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`AptpoTeNO-nrepdortiofx 31.02 Final version CONFIDENTIAL 04.12.2008 APPENDIX TO TNO-REPORT OF STUDY 3M-02 Project / study number: 031.10074/3M-02 TTNNOO QBuiaolsictiyenofceLsife GZae0umtiieBucosrxel2ea2tr1sas.tary Autre Th2P3re0o1NneeCthEse3rL1iaa7dn1ed5sn18 1200 CE4HHoPCaeagrtaeesn.mdaanNobel { Fax 13171518 1601 | In assignment of: I 3M, Medical Department || JStouhdynmBourtieonrhof,Ph.D. CiH, DABT `MCeodripcoarlatDeeSpcairetntmiesntt 3StMPaCueln,teMri,nBnueislodtiang505212404.-0120.00:02 UPhsoane: +1651733.1062 | For nes17RATI | E-mail jbutenhofi@cmomm `OrS.tuLdoyudiisreHc.toCr:ohen Status and Date: Final version: 04 December 2006 PDrreav"iou1s6vOerasticonbse:r2008 Dra. 30October2008 Numberofpages: 172 Authorization by Dr. P.H.A. Quax, Technology manoafVgasceulrar and Metabolic Diseases: Signature and date: \\ | S[/I|I 9 ( \N Page A1 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Contents Contents page A02 A. Code E3L mice before and after matching A03 B. Bodyweight and food intake measurements A04 B1. Individual bodyweights A04 B2. Food intake measurements A07 C. Plasma measurements A13 C1. Plasma cholesterol measurements A13 C2. Plasma triglyceride measurements A15 C3. Pooled lipoprotein profile measurements A17 C4. Pooled plasma ALAT measurements A24 C5. Plasma ketone bodies measurements A25 D. Measurements in metabolic cages A28 D1. Code mice and metabolic cages A28 D2. Average bodyweight mice in metabolic cages A28 D3. Food and drink intake measurements A29 D4. X activity measurements A38 D5. O2 consumption,CO2 production and respiratory exchange ratio measurements A41 D6. Energy expenditure A50 E. Fecal bile acid, sterol and fatty acid measurements A53 E1. Fecal weight measurements, bodyweight and food intake Measurements A53 E2. Fecal bile acid measurements A60 E3. Fecal sterol measurements A83 E4. Fecal fatty acid measurements A107 F. Intestinal cholesterol absorption A122 G. Liver weight, perigonadal fat weight and lipid contents liver G1. Liver weight and perigonadal fat weight G2. Lipid contents liver A127 A127 A129 H. HMG-CoA reductase and cholesterol-7-hydroxylase activity in liver microsomes and sterol-27-hydroxylase activity in liver mitochondria A136 H1. HMG-CoA reductase and cholesterol-7-hydroxylase activity in liver microsomes A136 H2. Sterol-27-hydroxylase activity in liver mitochondria A148 I. Measurements in vitro experiment (rat hepatocytes) A156 Page A2 of A172 Appendix to TNO-report of 3M-02 Final version Appendix A. Code E3L mice before and after matching mouse# cage# before matching 658 86 663 86 712 93 716 93 752 98 758 98 763 99 784 102 790 102 791 102 641 84 645 84 646 84 674 88 737 96 738 96 740 96 760 99 765 99 785 102 650 85 678 88 690 90/91 699 90/91 729 95 754 98 756 98 759 98 766 99 793 103 681 89 686 89 704 92 705 92 706 92 710 92 768 100 771 100 792 103 795 103 665 87 666 87 667 87 671 87 679 88 700 90/91 730 95 731 95 744 97 799 103 714 93 721 94 727 94 769 100 774 100 776 101 778 101 782 101 783 101 797 103 652 85 657 86 670 87 680 89 687 89 696 90/91 720 94 724 94 761 99 762 99 excluded (655) 85 excluded (656) 86 excluded (741) 96 excluded (742) 96 excluded (748) 97 eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 12 00 21 02 20 group# 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 5 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 mouse# after matching 1 1 2 2 3 3 4 5 5 5 6 6 6 7 8 8 8 9 9 10 11 12 13 13 14 15 15 15 16 17 18 18 19 19 19 19 20 20 21 21 22 22 22 22 23 24 25 25 26 27 28 29 29 30 30 31 31 31 31 32 33 34 35 36 36 37 38 38 39 39 cage# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 04-12-2006 Page A3 of A172 Appendix to TNO-report of 3M-02 Final version Appendix B. Bodyweight and food intake measurements B1. Individual bodyweights Group Mouse# cage# run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in run in AVERAGE SD run in run in run in run in run in 641 84 645 84 646 84 650 85 652 85 657 86 658 86 663 86 665 87 666 87 667 87 670 87 671 87 674 88 678 88 679 88 680 89 681 89 686 89 687 89 690 90/91 696 90/91 699 90/91 700 90/91 704 92 705 92 706 92 710 92 712 93 714 93 716 93 720 94 721 94 724 94 727 94 729 95 730 95 731 95 737 96 738 96 740 96 744 97 752 98 754 98 756 98 758 98 759 98 760 99 761 99 762 99 763 99 765 99 766 99 768 100 769 100 771 100 774 100 776 101 778 101 782 101 783 101 784 102 785 102 790 102 791 102 792 103 793 103 795 103 797 103 799 103 excluded (655) 85 excluded (656) 86 excluded (741) 96 excluded (742) 96 excluded (748) 97 eartag LR 10 21 02 01 20 10 01 12 10 01 11 02 12 01 02 12 00 10 02 12 01 00 11 20 00 10 01 02 00 01 20 00 10 20 12 10 01 11 10 01 20 00 00 01 20 02 12 00 10 01 11 21 02 00 10 11 02 00 01 02 22 00 10 02 12 00 10 11 21 12 12 00 21 02 20 bodyweight (g) week -2 week -1 week 0 30.0 30.1 29.2 27.5 28.1 28.7 31.2 30.9 30.6 27.5 27.6 28.5 29.9 30.0 31.1 31.4 30.9 32.2 32.6 31.8 31.9 30.1 30.9 31.1 30.5 30.7 31.3 25.0 25.6 25.4 28.0 27.7 27.9 30.7 31.0 31.3 28.6 27.9 29.9 31.5 31.4 31.9 29.9 30.0 30.3 29.3 29.0 30.9 31.4 31.5 31.5 29.4 29.2 30.9 29.1 28.9 29.1 29.9 29.6 31.5 31.4 31.0 31.8 26.2 26.3 26.7 27.6 27.9 27.8 29.7 29.4 29.5 29.0 29.7 29.2 31.4 30.9 30.4 32.0 31.9 31.8 24.6 24.8 26.4 29.4 29.1 29.8 29.0 29.6 30.2 28.8 28.4 28.9 28.2 28.8 29.1 32.5 32.6 31.8 27.5 27.1 29.0 26.5 26.9 28.0 25.0 25.3 27.0 29.4 29.4 30.7 28.8 28.5 29.3 28.9 29.0 30.0 28.1 28.7 28.3 30.6 30.9 30.9 27.0 27.4 27.7 29.0 29.4 30.5 28.8 29.0 31.1 28.2 28.0 29.0 27.8 27.3 29.3 24.5 24.7 26.1 25.2 25.7 26.4 23.1 23.8 24.0 25.5 25.9 25.5 25.4 25.0 25.9 27.2 26.8 26.5 28.0 28.1 27.9 27.0 27.6 28.9 28.1 28.2 30.4 29.0 29.3 29.8 27.2 27.1 28.0 25.5 25.7 25.3 28.2 28.3 28.1 27.5 27.6 29.0 27.1 27.0 29.5 28.6 28.9 28.9 29.3 29.7 28.8 28.8 28.7 29.0 26.1 26.8 26.6 27.4 27.5 26.0 27.7 27.8 27.9 26.8 26.7 26.9 29.0 29.5 28.0 29.1 29.4 29.5 28.43 28.51 29.03 2.03 1.93 1.93 27.3 27.0 28.5 22.2 22.9 24.6 29.0 29.2 30.9 31.0 31.6 32.2 30.6 30.9 31.0 04-12-2006 Page A4 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group Mouse# cage# 1 1 1 1 2 1 1 3 2 1 4 2 1 5 3 1 6 3 1 7 4 1 8 5 1 9 5 1 10 5 AVERAGE SD 2 11 6 2 12 6 2 13 6 2 14 7 2 15 8 2 16 8 2 17 8 2 18 9 2 19 9 2 20 10 AVERAGE SD 3 21 11 3 22 12 3 23 13 3 24 13 3 25 14 3 26 15 3 27 15 3 28 15 3 29 16 3 30 17 AVERAGE SD 4 31 18 4 32 18 4 33 19 4 34 19 4 35 19 4 36 19 4 37 20 4 38 20 4 39 21 4 40 21 AVERAGE SD 5 41 22 5 42 22 5 43 22 5 44 22 5 45 23 5 46 24 5 47 25 5 48 25 5 49 26 5 50 27 AVERAGE SD 6 51 28 6 52 29 6 53 29 6 54 30 6 55 30 6 56 31 6 57 31 6 58 31 6 59 31 6 60 32 AVERAGE SD 7 61 33 7 62 34 7 63 35 7 64 36 7 65 36 7 66 37 7 67 38 7 68 38 7 69 39 7 70 39 AVERAGE SD eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 week 0 31.9 31.1 29.8 28.9 30.5 29.3 25.9 28.9 29.0 26.6 29.19 1.85 29.2 28.7 30.6 31.9 30.0 28.3 30.9 26.4 26.5 28.8 29.13 1.80 28.5 30.3 31.8 27.8 27.0 31.1 29.0 26.1 27.9 27.9 28.74 1.82 30.9 29.1 29.2 30.4 31.8 26.4 28.9 29.8 26.0 26.9 28.94 1.95 31.3 25.4 27.9 29.9 30.9 29.5 30.7 29.3 27.7 29.5 29.21 1.78 30.2 31.8 28.0 30.4 28.0 25.3 28.1 29.0 29.5 28.0 28.83 1.79 31.1 32.2 31.3 31.5 31.5 26.7 29.1 29.0 24.0 25.5 29.19 2.88 week 1 31.4 29.8 30.2 29.4 30.0 28.4 25.8 29.7 28.9 26.2 28.98 1.76 29.4 28.5 30.7 31.7 30.2 28.2 31.6 25.9 26.3 28.1 29.06 2.03 27.4 29.2 32.4 27.8 26.3 30.8 30.1 26.4 27.5 27.8 28.57 2.00 30.5 29.3 28.6 31.0 31.6 25.6 28.3 30.3 27.5 26.1 28.88 2.04 29.5 24.7 26.9 29.4 29.4 28.4 27.9 27.9 26.3 28.0 27.84 1.53 29.2 31.2 27.1 30.8 28.0 24.8 27.8 27.7 29.8 27.5 28.39 1.91 30.1 30.2 28.8 30.6 30.4 26.2 28.2 28.4 23.4 25.2 28.15 2.46 week 2 32.6 29.8 29.8 28.6 30.9 27.8 24.8 29.0 29.8 26.3 28.94 2.24 29.8 28.9 30.3 32.3 29.7 29.4 31.7 25.9 26.5 28.8 29.33 2.00 27.3 29.7 32.0 28.6 26.1 30.0 29.9 25.3 27.8 27.4 28.41 2.03 30.2 29.2 29.0 31.6 31.4 25.6 29.1 30.7 28.0 26.3 29.11 2.02 25.6 23.8 26.8 25.4 26.9 26.7 28.3 25.8 24.0 26.1 25.94 1.36 28.8 31.7 27.6 30.9 28.4 24.9 28.1 28.4 30.4 27.2 28.64 1.98 27.0 26.6 26.6 28.9 28.4 25.0 26.6 26.0 22.4 23.6 26.11 2.00 week 2.3 25.0 23.8 26.6 25.0 27.0 26.5 28.4 25.6 24.6 25.3 25.78 1.35 26.9 26.4 25.2 27.7 27.3 24.5 25.6 25.4 21.7 23.6 25.43 1.82 week 3 32.5 29.8 29.6 27.9 30.5 28.1 25.0 28.6 29.7 26.2 28.79 2.15 30.2 29.0 29.3 31.7 30.2 29.6 31.7 25.6 26.8 28.2 29.23 1.96 27.1 28.9 32.6 28.8 25.6 29.1 29.9 24.9 27.8 27.3 28.20 2.20 29.7 29.2 28.9 30.8 31.1 25.7 28.3 30.2 27.4 27.0 28.83 1.74 25.6 24.8 27.0 25.7 27.9 27.9 28.4 28.0 25.4 27.0 26.77 1.30 28.7 31.4 27.1 30.5 28.1 25.1 27.7 28.2 30.0 27.1 28.39 1.85 26.2 25.0 24.7 27.8 27.1 24.6 25.5 25.3 22.3 23.5 25.20 1.61 week 3.4 28.0 24.8 27.3 27.5 29.1 28.6 29.5 29.7 25.2 27.6 27.73 1.67 26.0 24.1 25.0 27.7 26.8 25.2 26.2 26.0 23.1 24.3 25.44 1.37 week 4 32.6 29.9 29.8 27.9 30.4 27.6 25.7 28.1 29.2 26.1 28.73 2.08 31.0 28.9 29.4 31.9 30.2 29.4 31.8 26.4 27.0 28.3 29.43 1.87 27.8 29.0 32.5 29.2 25.2 29.2 30.0 25.0 28.9 26.7 28.35 2.27 29.7 28.9 28.9 30.9 31.1 25.3 28.2 30.6 27.5 27.3 28.84 1.83 27.5 24.2 26.7 28.1 28.9 28.4 29.2 29.3 25.7 27.8 27.58 1.63 28.3 31.3 27.1 30.3 28.8 25.1 27.9 28.9 30.6 27.2 28.55 1.87 26.3 24.7 25.3 27.9 27.1 25.3 25.8 26.1 22.8 23.8 25.51 1.50 Bodyweight (g) week 5 week 5.3 week 5.6 32.9 33.9 33.9 30.5 31.2 30.5 30.7 30.4 31.0 28.7 28.1 28.2 31.1 31.1 30.6 29.6 29.5 29.4 26.5 26.7 27.2 30.0 30.4 30.6 30.5 30.6 31.0 27.1 27.2 27.3 29.76 29.91 29.97 1.90 2.14 2.02 31.4 33.0 33.1 29.4 29.8 30.0 29.9 31.2 32.0 32.9 32.8 33.1 31.0 30.8 31.0 30.2 30.1 29.8 32.9 32.8 32.8 26.2 26.5 26.6 26.6 26.9 26.6 28.6 29.0 28.8 29.91 30.29 30.38 2.31 2.33 2.48 28.2 27.8 28.3 28.8 28.8 28.9 32.9 32.2 32.3 28.9 28.8 28.9 25.8 25.8 26.3 29.0 28.7 28.5 30.2 30.1 30.1 25.1 25.0 24.7 29.2 29.0 29.7 27.1 27.0 27.0 28.52 28.32 28.47 2.21 2.07 2.12 30.2 30.3 30.2 29.3 29.3 29.2 28.8 29.2 28.9 30.4 30.9 30.7 30.7 31.0 30.7 25.3 25.6 25.8 27.3 27.0 26.7 30.2 30.2 30.2 27.1 27.7 27.2 27.5 27.7 27.7 28.68 28.89 28.73 1.80 1.82 1.78 28.6 28.9 30.0 25.0 25.2 25.3 27.4 27.6 27.8 28.2 28.5 28.1 29.2 28.6 29.7 28.2 28.5 28.6 30.3 30.3 30.1 29.3 29.3 29.9 26.5 27.3 27.2 28.0 27.7 28.7 28.07 28.19 28.54 1.51 1.37 1.52 28.5 28.4 28.5 31.5 31.3 31.5 27.4 27.3 27.2 30.3 30.2 30.0 28.7 28.8 28.4 25.5 25.6 25.4 27.6 27.6 27.8 28.9 29.1 29.1 30.4 30.3 30.7 26.8 26.7 26.7 28.56 28.53 28.53 1.82 1.77 1.87 28.0 28.2 27.7 27.2 27.3 26.7 26.1 26.2 26.6 28.3 28.2 28.5 27.4 27.4 28.2 26.1 26.1 26.1 26.2 26.0 26.2 26.5 26.7 26.9 22.8 23.6 23.0 24.6 25.1 24.6 26.32 26.48 26.45 1.64 1.42 1.65 week 6 32.9 30.1 30.0 28.3 29.2 30.4 26.3 30.0 30.5 27.0 29.47 1.89 32.2 30.0 31.2 33.6 30.1 30.0 32.5 26.3 26.9 29.0 30.18 2.34 28.4 29.2 32.3 29.2 26.1 28.8 29.8 25.1 29.4 27.6 28.59 2.00 29.6 28.7 29.6 30.5 30.7 26.2 26.9 30.1 27.2 27.7 28.72 1.62 29.1 25.5 28.6 28.8 30.1 28.6 30.1 29.7 27.0 28.4 28.59 1.42 28.7 31.4 27.0 29.7 28.0 25.3 26.9 28.8 30.9 26.3 28.30 1.99 29.1 27.8 26.8 28.9 27.6 26.2 26.7 27.2 23.6 25.7 26.96 1.60 week 6.3 33.2 30.2 30.2 28.4 31.1 29.6 26.5 30.8 30.7 27.1 29.78 1.99 32.8 30.0 31.8 33.1 30.3 30.9 32.7 25.9 26.5 29.1 30.31 2.53 29.1 29.2 32.5 29.3 26.4 29.1 30.2 25.4 29.6 27.8 28.86 1.98 29.9 29.2 29.8 30.5 31.1 26.5 28.0 30.9 27.5 27.5 29.09 1.61 29.8 25.6 29.2 29.1 29.6 28.6 30.6 30.2 27.3 28.6 28.86 1.48 28.7 32.1 27.4 30.2 28.5 25.6 27.2 28.6 30.6 26.6 28.55 1.98 28.5 27.8 27.0 28.7 28.2 27.1 26.7 27.3 23.8 25.4 27.05 1.50 week 7 33.5 30.6 30.5 28.5 30.8 29.8 27.0 30.9 31.0 27.6 30.02 1.90 33.1 29.5 32.0 33.0 30.4 29.5 31.4 26.2 27.7 29.5 30.23 2.23 29.9 30.1 33.2 29.3 27.2 29.4 30.3 26.3 30.4 28.9 29.50 1.87 29.4 29.4 30.0 30.5 30.7 26.4 27.1 30.7 27.9 28.3 29.04 1.54 30.5 25.9 29.6 29.1 30.6 30.2 30.0 30.5 27.2 29.0 29.26 1.57 28.9 32.0 27.6 30.0 28.1 25.5 26.7 28.5 30.4 26.3 28.40 2.01 28.6 28.4 26.8 28.6 28.3 27.2 26.9 27.1 24.0 25.2 27.11 1.53 week 8 33.8 31.2 30.2 28.6 31.0 29.7 27.5 31.0 30.7 27.3 30.10 1.94 34.1 29.5 32.2 33.7 29.9 30.5 31.2 26.4 28.7 29.7 30.59 2.32 30.2 30.0 33.1 29.5 27.0 30.1 30.1 26.4 30.8 28.6 29.58 1.91 29.7 29.0 30.2 30.3 30.8 26.5 27.6 30.9 28.1 28.5 29.16 1.47 30.4 25.6 29.1 29.1 30.4 29.9 30.4 30.6 27.7 29.1 29.23 1.56 28.8 32.1 26.8 30.3 28.5 25.6 26.7 28.3 30.1 26.5 28.37 2.03 29.0 28.6 27.1 29.1 28.6 27.3 27.2 27.6 24.3 25.7 27.45 1.53 week 8.4 33.7 31.0 30.3 28.5 30.8 29.9 27.3 31.9 30.6 27.1 30.11 2.03 33.7 29.5 33.1 33.5 30.8 31.0 31.7 26.5 28.9 29.5 30.82 2.29 30.5 30.1 33.2 29.3 27.2 31.0 30.3 26.1 31.1 28.5 29.73 2.05 29.0 28.7 29.8 30.4 30.7 26.4 27.2 30.6 27.6 28.4 28.88 1.50 30.4 25.4 29.1 29.2 30.1 29.8 30.7 30.5 28.1 28.9 29.22 1.57 28.5 32.1 26.7 28.1 27.4 25.5 26.8 28.2 30.2 26.3 27.98 1.96 29.4 28.7 27.8 29.5 28.6 27.4 27.5 27.6 24.7 26.3 27.75 1.45 week 9 34.5 31.1 30.0 28.4 31.2 30.0 27.7 33.1 30.4 27.0 30.34 2.32 34.8 30.1 33.6 33.6 31.2 31.2 32.4 27.8 29.8 30.1 31.46 2.14 30.6 30.0 33.4 29.4 27.2 31.4 30.7 26.3 31.6 28.1 29.87 2.17 28.4 28.1 29.6 30.2 30.9 26.4 27.0 30.4 27.2 28.6 28.68 1.55 30.2 25.0 28.9 29.3 29.6 29.7 31.0 30.4 28.5 29.0 29.16 1.64 28.4 32.2 26.6 27.8 27.1 25.7 26.7 27.9 30.0 26.3 27.87 1.95 29.7 29.2 28.2 29.8 28.9 27.8 27.6 28.0 25.0 26.8 28.10 1.45 week 10 34.3 31.4 30.5 28.9 32.0 29.3 28.0 33.2 31.7 27.4 30.67 2.26 35.4 30.6 33.3 33.8 30.6 30.7 32.9 28.3 30.8 30.1 31.65 2.12 31.0 30.9 33.8 28.4 27.3 30.9 31.3 25.9 32.7 27.7 29.99 2.54 28.2 28.1 29.2 29.6 30.3 26.2 26.7 29.7 26.8 28.1 28.29 1.41 31.3 25.4 29.6 29.4 29.5 30.0 31.2 30.7 28.4 29.0 29.45 1.70 27.8 31.3 26.2 27.7 26.8 25.2 26.7 27.6 29.9 26.0 27.52 1.85 29.8 29.5 28.0 29.8 29.5 27.9 27.5 28.1 25.1 27.6 28.28 1.45 Page A5 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 week 0 1 2 3 4 5 5.3 5.6 6 6.3 7 8 8.4 9 10 week 0 1 2 3 4 5 5.3 5.6 6 6.3 7 8 8.4 9 10 week 0 1 2 2.3 3 3.4 4 5 5.3 5.6 6 6.3 7 8 8.4 9 10 group 1 vs 2 nn p 10 10 0.739 10 10 0.912 10 10 0.796 10 10 0.631 10 10 0.436 10 10 0.971 10 10 0.739 10 10 0.739 10 10 0.579 10 10 0.579 10 10 0.971 10 10 0.739 10 10 0.529 10 10 0.247 10 10 0.393 group 2 vs 4 nn p 10 10 0.912 10 10 0.853 10 10 0.796 10 10 0.684 10 10 0.529 10 10 0.280 10 10 0.247 10 10 0.190 10 10 0.165 10 10 0.218 10 10 0.218 10 10 0.165 10 10 0.035 10 10 0.009 10 10 <0.001 group 3 vs 7 nn p 10 10 0.481 10 10 0.971 10 10 0.023 10 10 0.003 10 10 0.011 10 10 0.029 10 10 0.043 10 10 0.015 10 10 0.052 10 10 0.023 10 10 0.003 10 10 0.019 10 10 0.043 10 10 0.052 10 10 0.123 group 1 vs 3 nn p 10 10 0.481 10 10 0.631 10 10 0.579 10 10 0.481 10 10 0.631 10 10 0.143 10 10 0.105 10 10 0.123 10 10 0.190 10 10 0.165 10 10 0.280 10 10 0.353 10 10 0.684 10 10 0.853 10 10 0.481 group 2 vs 5 nn p 10 10 0.796 10 10 0.143 10 10 0.001 10 10 0.005 10 10 0.029 10 10 0.043 10 10 0.035 10 10 0.075 10 10 0.089 10 10 0.075 10 10 0.039 10 10 0.190 10 10 0.075 10 10 0.015 10 10 0.035 group 4 vs 5 nn p 10 10 0.631 10 10 0.280 10 10 0.002 10 10 0.009 10 10 0.165 10 10 0.393 10 10 0.315 10 10 0.579 10 10 0.796 10 10 0.631 10 10 0.853 10 10 0.739 10 10 0.529 10 10 0.353 10 10 0.089 Bodyweight, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 group 1 vs 7 nn p nn p nn p nn p 10 10 0.853 10 10 0.796 10 10 0.529 10 10 0.631 10 10 0.971 10 10 0.075 10 10 0.393 10 10 0.631 10 10 0.796 10 10 0.004 10 10 0.631 10 10 0.011 10 10 0.912 10 10 0.019 10 10 0.739 10 10 0.001 10 10 0.912 10 10 0.218 10 10 0.912 10 10 0.002 10 10 0.247 10 10 0.029 10 10 0.165 10 10 0.001 10 10 0.218 10 10 0.063 10 10 0.143 10 10 0.002 10 10 0.143 10 10 0.075 10 10 0.165 10 10 <0.001 10 10 0.481 10 10 0.218 10 10 0.218 10 10 0.005 10 10 0.481 10 10 0.190 10 10 0.247 10 10 0.007 10 10 0.165 10 10 0.190 10 10 0.075 10 10 0.003 10 10 0.247 10 10 0.280 10 10 0.063 10 10 0.004 10 10 0.143 10 10 0.247 10 10 0.029 10 10 0.023 10 10 0.105 10 10 0.247 10 10 0.019 10 10 0.019 10 10 0.029 10 10 0.315 10 10 0.003 10 10 0.035 group 2 vs 6 group 2 vs 7 group 3 vs 4 group 3 vs 5 nn p nn p nn p nn p 10 10 0.579 10 10 0.579 10 10 0.739 10 10 0.529 10 10 0.353 10 10 0.481 10 10 0.631 10 10 0.853 10 10 0.353 10 10 0.005 10 10 0.393 10 10 0.007 10 10 0.280 10 10 <0.001 10 10 0.353 10 10 0.105 10 10 0.353 10 10 <0.001 10 10 0.684 10 10 0.393 10 10 0.247 10 10 0.001 10 10 0.631 10 10 0.631 10 10 0.123 10 10 0.001 10 10 0.393 10 10 0.796 10 10 0.105 10 10 0.002 10 10 0.579 10 10 0.912 10 10 0.075 10 10 0.004 10 10 0.684 10 10 1.000 10 10 0.105 10 10 0.009 10 10 0.579 10 10 0.912 10 10 0.089 10 10 0.003 10 10 0.912 10 10 0.853 10 10 0.052 10 10 0.002 10 10 0.853 10 10 0.853 10 10 0.011 10 10 0.002 10 10 0.315 10 10 0.436 10 10 0.002 10 10 <0.001 10 10 0.218 10 10 0.353 10 10 <0.001 10 10 <0.001 10 10 0.105 10 10 0.579 group 4 vs 6 group 4 vs 7 group 5 vs 6 group 5 vs 7 nn p nn p nn p nn p 10 10 0.796 10 10 0.631 10 10 0.684 10 10 0.631 10 10 0.529 10 10 0.529 10 10 0.684 10 10 0.393 10 10 0.529 10 10 0.007 10 10 0.002 10 10 0.796 10 10 0.853 10 10 0.579 10 10 <0.001 10 10 0.043 10 10 0.023 10 10 0.007 10 10 0.579 10 10 0.001 10 10 0.353 10 10 0.011 10 10 0.912 10 10 0.011 10 10 0.579 10 10 0.015 10 10 0.529 10 10 0.009 10 10 0.796 10 10 0.007 10 10 0.796 10 10 0.015 10 10 0.912 10 10 0.007 10 10 0.739 10 10 0.035 10 10 0.631 10 10 0.035 10 10 0.481 10 10 0.019 10 10 0.529 10 10 0.005 10 10 0.353 10 10 0.029 10 10 0.190 10 10 0.003 10 10 0.315 10 10 0.035 10 10 0.165 10 10 0.005 10 10 0.165 10 10 0.143 10 10 0.089 10 10 0.019 10 10 0.165 10 10 0.481 10 10 0.063 10 10 0.063 10 10 0.190 10 10 0.912 10 10 0.035 10 10 0.075 group 2 vs 3 nn p 10 10 0.529 10 10 0.481 10 10 0.353 10 10 0.190 10 10 0.247 10 10 0.143 10 10 0.052 10 10 0.089 10 10 0.075 10 10 0.105 10 10 0.393 10 10 0.436 10 10 0.280 10 10 0.165 10 10 0.436 group 3 vs 6 nn p 10 10 0.579 10 10 0.912 10 10 0.796 10 10 0.912 10 10 0.971 10 10 0.971 10 10 0.796 10 10 0.971 10 10 0.684 10 10 0.579 10 10 0.218 10 10 0.218 10 10 0.063 10 10 0.052 10 10 0.043 group 6 vs 7 nn p 10 10 0.481 10 10 0.912 10 10 0.015 10 10 0.001 10 10 0.002 10 10 0.009 10 10 0.015 10 10 0.019 10 10 0.190 10 10 0.105 10 10 0.247 10 10 0.631 10 10 0.853 10 10 0.436 10 10 0.218 p<0.05 Page A6 of A172 Appendix to TNO-report of 3M-02 B2. Food intake measurements Final version 04-12-2006 cage# 84 85 86 87 88 89 90/91 92 93 94 95 96 97 98 99 100 101 102 103 number of mice 3 3 4 5 3 4 4 4 3 4 3 5 2 5 6 4 4 4 5 Food on cage week -2 (g) 86.1 88.8 117.2 131.5 84.2 105.6 108.5 107.7 79.3 104.1 82.6 132.8 54.5 139.5 156.4 105.8 102.2 102.3 132.2 Food off cage week -1 (g) 37.1 41.6 49.1 51.8 22.9 29.3 40.9 49.6 21 40 25.7 40 20.8 57.2 78.2 30.5 40.3 44.7 53.6 days 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 Food intake (g/day/mouse) 2.72 2.62 2.84 2.66 3.41 3.18 2.82 2.42 3.24 2.67 3.16 3.09 2.81 2.74 2.17 3.14 2.58 2.40 2.62 2.80 0.32 Food on cage week -1 (g) 82 81.8 108.9 132.7 77.6 103.9 105.2 108.2 80.3 108.8 81.6 130.8 52.2 129.7 152.1 104.2 106.9 106.8 130.6 Food off cage week 0 (g) 25 19.7 30.3 39.4 10.4 20.9 27.7 40.3 16.7 31.1 17.1 28.5 9.5 33.2 51.8 20.5 31 33.2 41.7 days 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 Food intake (g/day/mouse) 2.71 2.96 2.81 2.67 3.20 2.96 2.77 2.43 3.03 2.78 3.07 2.92 3.05 2.76 2.39 2.99 2.71 2.63 2.54 2.81 0.22 Page A7 of A172 Page A8 of A172 Group 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 AVERAGE SD cage# 1 2 3 4 5 number of mice 2 2 2 1 3 Food on cage week 0 (g) 70.8 69 62.5 31 86.1 Food off cage week 1 (g) 34.8 26.5 26.4 12.5 38.3 days 6 6 6 6 6 6 3 85.8 34.1 6 7 1 38.3 15.3 6 8 3 89.4 31.4 6 9 2 59.8 28.6 6 10 1 34.1 18.2 6 11 1 34.4 17 6 12 1 35 16.4 6 13 2 63 24.3 6 14 1 32.2 13.5 6 15 3 84.1 31.3 6 16 1 29 12.6 6 17 1 30.9 12.2 6 18 2 19 4 20 2 21 2 62.4 110.6 54.5 65.1 20.9 6 47.6 6 12.5 6 29.8 6 22 4 23 1 24 1 25 2 26 1 27 1 108.8 29.5 29.6 53.8 32 32 48.9 6 10.7 6 11.6 6 17.5 6 15.9 6 14.9 6 28 1 29 2 30 2 31 4 32 1 36.1 59.2 60.2 108.1 40.5 16 6 20.1 6 20.7 6 44.7 6 23.6 6 33 1 33.3 14.2 6 34 1 33.8 14.8 6 35 1 40.8 23.1 6 36 2 56.5 16.1 6 37 1 30.8 14.5 6 38 2 50.5 18.1 6 39 2 55.7 25.1 6 Food intake g/day/mouse 3.00 3.54 3.01 3.08 2.66 3.06 0.32 2.87 3.83 3.22 2.60 2.65 3.04 0.51 2.90 3.10 3.23 3.12 2.93 2.73 3.12 3.02 0.17 3.46 2.63 3.50 2.94 3.13 0.42 2.50 3.13 3.00 3.03 2.68 2.85 2.86 0.24 3.35 3.26 3.29 2.64 2.82 3.07 0.32 3.18 3.17 2.95 3.37 2.72 2.70 2.55 2.95 0.30 Food on cage week 1 (g) 51.4 52.3 51.1 29.7 83.6 Food off cage week 2 (g) 12 9.9 10.9 10.5 31.4 80.7 24.4 28.5 4.3 81.3 20.1 52.9 18 31.1 12.1 26.4 6.1 31.4 11.6 60 16.2 27.6 7.8 76.3 23.8 27.9 8.2 28.3 8.5 50.9 11.4 98.6 33.7 48.6 8.1 56.8 17.8 100.7 40.6 24.9 5.6 27.2 11.2 49.2 14.4 25.6 9 29.5 13 29 8.6 53.8 11.8 51.2 9.3 101.5 33 31.1 11.1 30.5 12.8 27.6 11.9 28.2 10.8 55.6 21 29.2 13.5 54.2 25.7 51.2 21.4 days 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 Food intake g/day/mouse 3.28 3.53 3.35 3.20 2.90 3.25 0.23 3.13 4.03 3.40 2.91 3.17 3.33 0.43 3.38 3.30 3.65 3.30 2.92 3.28 3.30 3.30 0.21 3.29 2.70 3.38 3.25 3.16 0.31 2.50 3.22 2.67 2.90 2.77 2.75 2.80 0.24 3.40 3.50 3.49 2.85 3.33 3.32 0.27 2.95 2.62 2.90 2.88 2.62 2.38 2.48 2.69 0.22 Food on cage week 2 (g) 57.2 55 55.7 27.6 75.7 Food off cage week 2.3 days 77.6 27 83.1 55.5 31.8 29.9 29.4 56.3 30.5 81.6 35.1 30.1 57.4 106.4 53.4 52.3 108.7 31.7 31.3 57.8 30.3 34.1 83.2 2.8 23.7 2.8 24.3 2.8 41.4 2.8 23 2.8 27.4 2.8 35.1 57.4 50.2 104.6 36 37.6 30.8 2.8 32.4 24.6 2.8 31.1 24.8 2.8 54.6 41 2.8 33.9 28.1 2.8 55.7 44.7 2.8 56.4 44.5 2.8 Food intake g/day/mouse 2.28 2.86 2.50 2.93 2.61 2.39 2.59 0.26 2.43 2.79 2.25 2.43 2.07 1.96 2.13 2.29 0.28 Food on cage week 2.3 Food off cage week 3 (g) 15.2 9.3 19.2 5.1 17.4 8.7 9.6 16.5 17.3 10.2 17.5 7.9 8.4 10.3 25.2 15.3 10.1 15.7 24.9 10 11.6 108.6 62.5 27.2 14.9 31.8 20.1 57.1 31.1 29.5 15.5 31.0 17.8 14.3 10.3 7.1 25.2 17.8 32.9 23 33.6 24.2 27.6 16.6 52.5 26.7 28.7 18.5 54.9 34.5 60.0 26.7 days 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 4.2 4.2 4.2 4.2 4.2 4.2 7 7 7 7 7 4.2 4.2 4.2 4.2 4.2 4.2 4.2 Food intake g/day/mouse 3.00 3.26 2.61 3.21 2.78 2.97 0.28 3.28 2.49 3.17 2.73 3.09 2.95 0.33 1.77 3.07 3.42 2.89 2.69 2.83 2.86 2.94 0.28 2.98 2.91 3.10 2.91 2.97 0.09 2.74 2.93 2.79 3.10 3.33 3.14 3.00 0.23 2.97 3.36 3.08 2.84 2.60 2.97 0.28 2.36 2.24 2.62 3.07 2.43 2.43 3.96 2.73 0.61 Food on cage week 3 (g) 64.6 59.6 59.6 30.6 80.4 Food off cage week 4 (g) 28.8 21 21.2 10.7 30.8 days 6 6 6 6 6 77.4 19.5 6 30.1 8.4 6 79 18.5 6 54.3 19.6 6 33.5 14 6 31.3 11 6 32.8 13.7 6 67.6 24.8 6 27.4 9.1 6 78.4 26.5 6 32.9 14 6 30.2 11.5 6 56.7 110.8 55.6 59.1 18.7 6 46.9 6 16.2 6 23.1 6 104.1 38.5 40.3 65.1 35.3 31.3 30.4 6 22.5 6 19.7 6 21.8 6 18.8 6 12.3 6 29 54.5 54.8 105.1 29.5 9.5 6 16.3 6 19.1 6 39 6 11.1 6 50.1 32.1 6 41.5 23.3 6 31.5 13.4 6 60.4 28.1 6 27.3 8.2 6 52.7 18.4 6 58.5 28.1 6 Food intake g/day/mouse 2.98 3.22 3.20 3.32 2.76 3.09 0.23 3.22 3.62 3.36 2.89 3.25 3.27 0.26 3.38 3.18 3.57 3.05 2.88 3.15 3.12 3.15 0.25 3.17 2.66 3.28 3.00 3.03 0.27 3.07 2.67 3.43 3.61 2.75 3.17 3.12 0.37 3.25 3.18 2.98 2.75 3.07 3.05 0.19 3.00 3.03 3.02 2.69 3.18 2.86 2.53 2.90 0.22 04-12-2006 Final version Appendix to TNO-report of 3M-02 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group cage# 1 1 1 2 1 3 1 4 1 5 AVERAGE SD 2 6 2 7 2 8 2 9 2 10 AVERAGE SD 3 11 3 12 3 13 3 14 3 15 3 16 3 17 AVERAGE SD 4 18 4 19 4 20 4 21 AVERAGE SD 5 22 5 23 5 24 5 25 5 26 5 27 AVERAGE SD 6 28 6 29 6 30 6 31 6 32 AVERAGE SD 7 33 7 34 7 35 7 36 7 37 7 38 7 39 AVERAGE SD number of mice 2 2 2 1 3 3 1 3 2 1 1 1 2 1 3 1 1 2 4 2 2 4 1 1 2 1 1 1 2 2 4 1 1 1 1 2 1 2 2 Food on cage week 4 (g) 50.3 55.6 51.7 26.2 83.0 Food off cage week 5 (g) 19.0 22.0 18.3 10.0 34.6 days 5.5 5.5 5.5 5.5 5.5 74.4 26.0 5.5 29.7 10.8 5.5 75.5 24.6 5.5 50.2 23.8 5.5 25.7 9.5 5.5 26.5 9.8 5.5 29.5 14.7 5.5 55.8 18.2 5.5 30.2 16.1 5.5 84.8 42.1 5.5 25.7 10.0 5.5 35.6 20.9 5.5 53.0 107.9 58.2 58.0 19.4 5.5 52.0 5.5 26.7 5.5 29.5 5.5 105.0 30.5 30.2 53.0 28.5 27.6 60.2 4.5 16.8 4.5 15.7 4.5 22.2 4.5 15.4 4.5 14.3 4.5 30.2 50.5 54.1 102.0 30.0 16.3 5.5 18.2 5.5 22.6 5.5 45.9 5.5 17.1 5.5 33.2 17.4 4.5 31.4 16.7 4.5 41.8 29.1 4.5 45.3 16.2 4.5 26.2 13.8 4.5 65.5 35.9 4.5 55.0 33.8 4.5 Food intake g/day/mouse 2.85 3.05 3.04 2.95 2.93 2.96 0.08 2.93 3.44 3.08 2.40 2.95 2.96 0.37 3.04 2.69 3.42 2.56 2.59 2.85 2.67 2.83 0.31 3.05 2.54 2.86 2.59 2.76 0.24 2.49 3.04 3.22 3.42 2.91 2.96 3.01 0.32 2.53 2.94 2.86 2.55 2.34 2.64 0.25 3.51 3.27 2.82 3.23 2.76 3.29 2.36 3.03 0.40 Food on cage week 5 (g) 60.2 60.7 57.2 31.6 77.1 Food off cage week 6 (g) 22.1 19.5 16.8 10.7 19.2 85.4 19.8 37.4 12.4 85.2 14.2 51.9 13.6 33.6 13.4 36 15.7 37.4 16.2 55.9 7.1 35.4 15.6 89.8 32.7 27.4 8 31.2 9.4 57.7 16.5 106.9 33.2 53.4 11.1 57.7 17.7 109.2 32.3 41.4 20.1 32.5 10.6 61.1 15.5 32.8 12.5 31.6 10.2 36.3 16.6 60.8 20.3 52.6 12.3 105.5 36.7 34.8 16 33.2 12.7 35.4 16.3 39.9 20 78.1 38.8 31.1 12.1 61.2 24.9 52.9 15 days 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 Food intake g/day/mouse 2.72 2.94 2.89 2.99 2.76 2.86 0.12 3.12 3.57 3.38 2.74 2.89 3.14 0.34 2.90 3.03 3.49 2.83 2.72 2.77 3.11 2.98 0.26 2.94 2.63 3.02 2.86 2.86 0.17 2.75 3.04 3.13 3.26 2.90 3.06 3.02 0.18 2.81 2.89 2.88 2.46 2.69 2.75 0.18 2.93 2.73 2.84 2.81 2.71 2.59 2.71 2.76 0.11 Food on cage week 6 (g) 78.3 73.4 67 38.3 94.8 Food off cage week 7 (g) 61.7 55.5 48.9 29.1 71 days 3 3 3 3 3 97.1 69.2 3 42 32.6 3 85.6 54.1 3 60.9 46.3 3 46.3 37.3 3 39.9 40.9 56.5 46.1 114.5 34.9 37.9 30.3 3 31.4 3 39.2 3 37.8 3 91.4 3 26.2 3 29.5 3 61.5 130.6 70 67.9 42.6 3 98.9 3 50.3 3 50.3 3 138.4 52.3 37.3 74.9 39.9 35.4 107.2 3 44.3 3 27.6 3 53.8 3 31.1 3 26.8 3 44.3 34.9 3 66.8 48.7 3 60 42.9 3 142 112.9 3 46.7 39 3 35.7 27.4 3 42.4 33.8 3 46.1 36.9 3 90.6 73.3 3 39.7 30.7 3 76.5 60.4 3 65.9 50.8 3 Food intake g/day/mouse 2.77 2.98 3.02 3.07 2.64 2.90 0.18 3.10 3.13 3.50 2.43 3.00 3.03 0.39 3.20 3.17 2.88 2.77 2.57 2.90 2.80 2.90 0.22 3.15 2.64 3.28 2.93 3.00 0.28 2.60 2.67 3.23 3.52 2.93 2.87 2.97 0.35 3.13 3.02 2.85 2.43 2.57 2.80 0.30 2.77 2.87 3.07 2.88 3.00 2.68 2.52 2.83 0.19 Page A9 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group cage# mouse# Food on cage Food off cage days Food intake Food on cage week 7 (g) week 7.5 (g) g/day/mouse week 7.5(g) 1 1 1 438.1 428.4 3 3.22 29.8 1 MC 101 2 metabolic cage 20.3 1 MC 105 3 metabolic cage 18.6 1 MC 201 4 metabolic cage 23.5 1 3 5 485.4 476.5 3 2.96 26.1 1 MC 206 6 metabolic cage 19.2 1 4 7 496.1 3 2.86 29.2 1 5 8,9,10 425.4 397.3 3 3.12 54.7 AVERAGE 3.04 SD 0.16 2 6 11,12,13 435 407.6 3 3.04 55.9 2 7 14 429.7 421.0 3 2.91 21.9 2 8 15,16,17 475.5 442.1 3 3.71 44.3 2 9 18,19 450.2 431.3 3 3.15 33.6 2 10 20 425.7 9.5 3 2.94 32.1 AVERAGE 3.15 SD 0.33 3 MC 102 21 metabolic cage 35.6 3 MC 106 22 metabolic cage 29.4 3 13 23 455.8 445.8 3 3.35 33.1 3 MC 202 24 metabolic cage 26.8 3 14 25 430.2 418.8 3 3.79 36.9 3 15 26,28 465.3 449.8 3 2.58 45.1 3 MC 206 27 metabolic cage 32.9 3 16 29 437.8 428.5 3 3.10 26.3 3 17 30 429.1 421.5 3 2.53 44.1 AVERAGE 3.07 SD 0.53 4 MC 103 31 metabolic cage 44.3 4 MC 107 32 metabolic cage 51.2 4 19 33,34,35,36 486.1 448.3 3 3.15 84.3 4 MC 203 37 metabolic cage 33.1 4 MC207 38 metabolic cage 32.6 4 21 39,40 436.2 419.7 3 2.75 44.2 AVERAGE 2.95 SD 0.28 5 22 41,42,43,44 505.9 470.3 3 2.97 109.2 5 23 45 443.9 434.1 3 3.26 41.4 5 24 46 438.7 428.7 3 3.35 32.5 5 25 47,48 443.3 423.9 3 3.24 61.1 5 26 49 428.6 419.7 3 2.98 32.8 5 27 50 436.1 427.5 3 2.86 31.6 AVERAGE 3.11 SD 0.20 6 MC104 51 metabolic cage 38.2 6 29 52 432.6 424.2 3 2.81 27.6 6 MC 108 53 metabolic cage 33.1 6 MC204 54 metabolic cage 28.4 6 MC208 55 metabolic cage 33.1 6 31 56,57,58,59 409.8 371.4 3 3.20 87.4 6 32 60 441.6 433.5 3 2.69 21.7 AVERAGE 2.90 SD 0.27 7 33 61 439.2 429.8 3 3.12 33.1 7 34 62 431.4 422.0 3 3.15 32.5 7 35 63 442.7 434.7 3 2.67 39.4 7 36 64,65 445.6 427.0 3 3.10 66.1 7 37 66 426.1 417.7 3 2.79 30.2 7 38 67,68 465.5 446.0 3 3.25 65.8 7 39 69,70 445.2 423.9 3 3.55 52.1 AVERAGE 3.09 SD 0.29 NB: week 7-7.5: all mice are put on powdered food because of metabolic cages of part of mice Food off cage week 8 (g) 15.7 12.8 8.7 15.0 13.9 8.3 18.2 25.8 12.5 11.9 11.7 8.4 17.3 26.2 21.4 20.3 17.7 24.7 20.5 25.1 15.5 34.7 35.6 43.3 36.0 23.1 23.2 27.0 63.4 31.1 18.9 38.2 20.9 22.2 27.5 15.6 25.6 20.7 24.7 46.3 11.4 22.4 22.5 27.5 39.3 18.8 40.7 32.5 days 4 3 3 3 4 3 4 4 4 4 4 4 4 3 3 4 3 4 4 3 4 4 3 3 4 3 3 4 4 4 4 4 4 4 3 4 3 3 3 4 4 4 4 4 4 4 4 4 Food intake g/day/mouse 3.52 2.51 3.29 2.84 3.05 3.63 2.75 2.41 3.00 0.45 3.62 2.51 2.72 3.15 3.70 3.14 0.53 3.12 2.67 3.20 3.02 3.06 3.08 2.61 2.71 2.36 2.87 0.29 2.89 2.62 3.02 3.34 3.14 2.15 2.86 0.42 2.86 2.57 3.41 2.86 2.98 2.36 2.84 0.36 3.58 3.00 2.49 2.57 2.81 2.57 2.58 2.80 0.39 2.68 2.51 2.98 3.35 2.84 3.14 2.45 2.85 0.33 Page A10 of A172 Appendix to TNO-report of 3M-02 Final version Group Mouse# 1 1 1 2 1 3 1 4 1 5 1 6 1 7 1 8 1 9 1 10 AVERAGE SD 2 11 2 12 2 13 2 14 2 15 2 16 2 17 2 18 2 19 2 20 AVERAGE SD 3 21 3 22 3 23 3 24 3 25 3 26 3 27 3 28 3 29 3 30 AVERAGE SD 4 31 4 32 4 33 4 34 4 35 4 36 4 37 4 38 4 39 4 40 AVERAGE SD 5 41 5 42 5 43 5 44 5 45 5 46 5 47 5 48 5 49 5 50 AVERAGE SD 6 51 6 52 6 53 6 54 6 55 6 56 6 57 6 58 6 59 6 60 AVERAGE SD 7 61 7 62 7 63 7 64 7 65 7 66 7 67 7 68 7 69 7 70 AVERAGE SD cage# 1 1 2 2 3 3 4 5 5 5 6 6 6 7 8 8 8 9 9 10 11 12 13 13 14 15 15 15 16 17 18 18 19 19 19 19 20 20 21 21 22 22 22 22 23 24 25 25 26 27 28 29 29 30 30 31 31 31 31 32 33 34 35 36 36 37 38 38 39 39 Food on cage week 8 (g) 34.3 27.1 28.6 44.1 36.9 43.3 42.2 34.2 33.0 32.9 Food off cage week 9 (g) 11.7 5.8 7.9 24.6 15.6 20.5 22.2 10.1 12.1 8.6 days 7 7 7 7 7 7 7 7 7 7 34.1 9.5 7 36.5 16.4 7 28.4 6.3 7 29.5 11.4 7 28.3 5.8 7 27.1 3.2 7 36.0 15.9 7 44.3 22.2 7 45.2 25.0 7 33.9 9.3 7 34.0 12.0 7 35.1 14.2 7 29.8 7.7 7 27.6 8.2 7 44.1 18.8 7 44.3 23.4 7 47.2 26.3 7 34.9 13.8 7 33.2 13.1 7 32.6 12.6 7 33.1 11.5 7 34.2 15.5 7 45.7 24.8 7 36.0 14.0 7 45.7 23.0 7 44.1 25.6 7 36.9 17.2 7 32.1 10.7 7 22.5 1.2 7 33.4 12.3 7 44.1 19.5 7 37.7 18.9 7 45.1 21.2 7 44.2 25.6 7 35.6 15.2 7 43.2 19.8 7 44.8 26.1 7 42.5 18.6 7 29.6 7.9 7 34.3 15.6 7 33.9 10.7 7 32.4 14.6 7 35.6 18.6 7 41.8 19.9 7 42.1 22.0 7 44.0 23.1 7 43.3 23.1 7 32.5 9.8 7 28.2 7.1 7 29.4 12.0 7 34.4 14.3 7 45.2 21.3 7 30.1 8.8 7 29.8 7.7 7 27.2 3.8 7 33.4 12.5 7 31.5 10.2 7 46.2 26.1 7 44.1 23.2 7 32.5 14.4 7 Food intake g/day/mouse 3.23 3.05 2.96 2.79 3.05 3.26 2.86 3.45 2.98 3.47 3.11 0.23 3.52 2.87 3.16 2.58 3.22 3.42 2.87 3.16 2.88 3.52 3.12 0.32 3.15 2.98 3.16 2.77 3.62 2.99 2.99 3.01 2.87 2.86 3.04 0.24 3.08 2.67 2.98 3.15 3.24 2.65 2.81 3.06 3.04 3.02 2.97 0.20 3.52 2.68 3.42 2.66 2.91 3.35 2.67 3.42 3.10 2.67 3.04 0.36 3.32 2.55 2.43 3.13 2.87 2.98 2.88 3.24 3.01 2.49 2.89 0.31 2.87 3.41 3.05 3.16 3.34 2.99 3.04 2.87 2.99 2.58 3.03 0.24 04-12-2006 Page A11 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 week 0-1 1-2 2-3 (gr 5,7: 2.3-3) 3-4 4-5 5-6 6-7 7-7.5 7.5-8 8-9 week 0-1 1-2 2-3 (gr 5,7: 2.3-3) 3-4 4-5 5-6 6-7 7-7.5 7.5-8 8-9 week 0-1 1-2 2-2.3 2-3 (gr 5,7: 2.3-3) 3-4 4-5 5-6 6-7 7-7.5 7.5-8 8-9 group 1 vs 2 nn p 5 5 0.690 5 5 1.000 5 5 1.000 5 5 0.222 5 5 0.841 5 5 0.222 5 5 0.310 4 5 0.905 8 5 0.622 10 10 0.912 group 2 vs 4 nn p 5 4 0.730 5 4 0.905 5 4 0.905 5 4 0.286 5 4 0.413 5 4 0.286 5 4 1.000 5 2 0.571 5 6 0.429 10 10 0.247 group 3 vs 7 nn p 7 7 0.805 7 7 0.001 group 1 vs 3 nn p 5 7 0.876 5 7 0.530 5 7 0.755 5 7 0.876 5 7 0.202 5 7 0.432 5 7 1.000 4 5 1.000 8 9 0.606 10 10 0.529 group 2 vs 5 nn p 5 6 0.792 5 6 0.030 5 6 0.792 5 6 0.429 5 6 0.931 5 6 0.792 5 6 0.792 5 6 0.931 5 6 0.429 10 10 0.631 group 4 vs 5 nn p 4 6 0.476 4 6 0.114 Food intake, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 nn p nn p nn p 5 4 0.905 5 6 0.429 5 5 1.000 5 4 0.905 5 6 0.017 5 5 0.690 5 4 0.905 5 6 0.931 5 5 1.000 5 4 0.730 5 6 0.931 5 5 0.548 5 4 0.286 5 6 0.662 5 5 0.056 5 4 0.222 5 6 0.126 5 5 0.222 5 4 0.310 5 6 1.000 5 5 0.690 4 2 0.800 4 6 0.352 4 3 0.400 8 6 0.755 8 6 0.662 8 7 0.463 10 10 0.353 10 10 0.579 10 10 0.190 group 2 vs 6 group 2 vs 7 group 3 vs 4 nn p nn p nn p 5 5 0.690 5 7 1.000 7 4 0.648 5 5 0.690 5 7 0.005 7 4 0.315 5 5 1.000 5 7 0.149 7 4 0.230 5 5 0.151 5 7 0.030 7 4 0.527 5 5 0.151 5 7 0.876 7 4 0.788 5 5 0.056 5 7 0.030 7 4 0.648 5 5 0.421 5 7 0.149 7 4 0.527 5 3 0.393 5 7 1.000 5 2 1.000 5 7 0.268 5 7 0.268 9 6 0.955 10 10 0.165 10 10 0.436 10 10 0.971 group 4 vs 6 group 4 vs 7 group 5 vs 6 nn p nn p nn p 4 5 0.730 4 7 0.527 6 5 0.329 4 5 0.190 4 7 0.042 6 5 0.017 7 7 0.383 4 6 0.762 4 5 0.905 4 7 0.230 6 5 7 7 0.026 4 6 0.610 4 5 1.000 4 7 0.527 6 5 7 7 0.318 4 6 0.352 4 5 0.413 4 7 0.315 6 5 7 7 0.073 4 6 0.171 4 5 0.413 4 7 0.230 6 5 7 7 0.535 4 6 0.762 4 5 0.286 4 7 0.315 6 5 5 7 0.755 2 6 0.429 2 3 1.000 2 7 0.667 6 3 9 7 0.918 6 6 0.699 6 7 0.445 6 7 0.836 6 7 10 10 0.684 10 10 0.631 10 10 0.529 10 10 0.739 10 10 0.931 0.931 0.082 0.030 0.429 0.167 0.836 0.315 group 1 vs 7 nn p 5 7 0.755 5 7 0.005 5 7 0.202 5 7 0.202 5 7 0.755 5 7 0.149 5 7 0.639 4 7 0.788 8 7 0.613 10 10 0.631 group 3 vs 5 nn p 7 6 0.295 7 6 0.002 7 6 0.445 7 6 0.836 7 6 0.366 7 6 0.445 7 6 0.731 5 6 0.931 9 6 0.607 10 10 0.796 group 5 vs 7 nn p 6 7 0.534 6 7 0.445 6 7 0.073 6 7 0.101 6 7 0.295 6 7 0.836 6 7 0.014 6 7 0.628 6 7 0.836 6 7 0.945 10 10 0.853 group 2 vs 3 nn p 5 7 0.639 5 7 0.755 5 7 0.639 5 7 0.432 5 7 0.432 5 7 0.432 5 7 0.530 5 5 1.000 5 9 0.298 10 10 0.436 group 3 vs 6 nn p 7 5 0.530 7 5 0.432 7 5 0.639 7 5 0.432 7 5 0.268 7 5 0.149 7 5 0.530 5 3 1.000 9 7 0.299 10 10 0.481 group 6 vs 7 nn p 5 7 0.530 5 7 0.018 5 7 0.202 5 7 0.268 5 7 0.149 5 7 1.000 5 7 1.000 3 7 0.667 7 7 0.805 10 10 0.315 p<0.05 Page A12 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Appendix C. Plasma measurements C1. Plasma cholesterol measurements group Mouse# cage# 1 1 1 1 2 1 1 3 2 1 4 2 1 5 3 1 6 3 1 7 4 1 8 5 1 9 5 1 10 5 AVERAGE SD 2 11 6 2 12 6 2 13 6 2 14 7 2 15 8 2 16 8 2 17 8 2 18 9 2 19 9 2 20 10 AVERAGE SD 3 21 11 3 22 12 3 23 13 3 24 13 3 25 14 3 26 15 3 27 15 3 28 15 3 29 16 3 30 17 AVERAGE SD 4 31 18 4 32 18 4 33 19 4 34 19 4 35 19 4 36 19 4 37 20 4 38 20 4 39 21 4 40 21 AVERAGE SD 5 41 22 5 42 22 5 43 22 5 44 22 5 45 23 5 46 24 5 47 25 5 48 25 5 49 26 5 50 27 AVERAGE SD 6 51 28 6 52 29 6 53 29 6 54 30 6 55 30 6 56 31 6 57 31 6 58 31 6 59 31 6 60 32 AVERAGE SD 7 61 33 7 62 34 7 63 35 7 64 36 7 65 36 7 66 37 7 67 38 7 68 38 7 69 39 7 70 39 AVERAGE SD excluded 655 85 excluded 656 86 excluded 741 96 excluded 742 96 excluded 748 97 eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 12 00 21 02 20 Measured cholesterol (mmol/l) week 0 week 1 week 4 week 8 3.219 2.814 2.172 2.211 3.446 3.088 2.599 2.394 3.633 3.176 2.120 1.725 3.260 2.556 1.911 1.877 1.847 2.612 2.042 1.733 3.819 3.458 2.033 2.500 4.055 3.958 1.815 2.166 3.949 3.668 2.347 2.295 4.590 3.257 1.972 1.816 3.706 3.015 1.911 1.854 3.998 1.791 4.428 4.274 4.453 1.774 1.750 4.079 4.055 3.633 3.901 2.145 3.281 2.878 3.080 2.306 2.322 3.450 2.959 3.233 2.625 1.110 2.094 1.859 2.443 1.057 1.563 2.233 2.233 2.085 2.181 0.867 2.287 1.353 2.272 0.837 1.209 2.151 2.439 2.029 4.201 3.446 4.047 4.071 3.649 3.893 3.130 3.519 4.290 2.083 3.217 3.007 3.257 2.999 2.685 2.741 2.798 3.201 3.466 1.968 1.850 1.876 2.077 1.920 1.406 1.885 1.737 1.850 2.521 1.153 2.439 2.021 1.771 2.318 1.763 1.870 1.703 2.219 2.431 1.315 3.974 3.625 4.104 3.122 3.698 1.701 3.576 3.511 3.917 2.010 1.565 1.622 2.057 1.912 2.322 1.904 1.895 1.879 2.467 1.557 1.327 1.327 1.476 1.284 1.380 1.301 1.467 1.354 1.754 1.293 1.125 1.353 1.422 1.186 1.490 1.209 1.452 1.156 1.672 1.429 1.661 3.617 3.682 3.795 4.323 3.901 3.641 3.065 1.669 2.708 1.871 1.525 1.839 1.670 3.056 2.226 1.485 1.638 2.008 2.274 1.179 1.197 1.162 1.188 1.258 1.388 0.813 1.449 1.354 1.467 1.072 1.885 1.983 1.490 2.591 2.454 0.746 1.429 1.247 2.097 2.862 3.138 4.086 3.316 3.511 3.584 1.523 3.795 3.357 3.365 1.992 2.604 2.556 2.258 2.492 2.508 1.654 2.330 2.024 2.065 1.101 1.101 1.223 1.110 1.136 1.110 1.214 1.005 1.249 1.310 1.794 1.247 1.953 1.611 1.543 1.968 0.928 1.452 1.763 2.386 3.462 3.763 3.024 3.479 3.324 3.357 4.241 4.274 3.917 3.795 1.646 1.775 1.541 1.549 1.501 1.581 1.879 1.928 1.839 1.783 2.338 3.174 2.573 2.242 1.920 2.077 1.998 1.963 1.824 1.894 2.113 2.021 2.447 2.280 1.816 1.832 1.733 2.158 1.695 1.505 6.230 1.336 1.360 0.792 0.776 week 0 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 dilution week 1 week 4 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 week 8 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 week 0 6.438 6.892 7.266 6.520 3.694 7.638 8.110 7.898 9.180 7.412 7.10 1.45 7.996 3.582 8.856 8.548 8.906 3.548 3.500 8.158 8.110 7.266 6.85 2.33 8.402 6.892 8.094 8.142 7.298 7.786 6.260 7.038 8.580 4.166 7.27 1.31 7.948 7.250 8.208 6.244 7.396 3.402 7.152 7.022 7.834 4.020 6.65 1.65 3.322 7.234 7.364 7.590 8.646 7.802 7.282 6.130 3.338 5.416 6.41 1.85 5.724 6.276 8.172 6.632 7.022 7.168 3.046 7.590 6.714 6.730 6.51 1.39 6.924 7.526 6.048 6.958 6.648 6.714 8.482 8.548 7.834 7.590 7.33 0.81 12.460 2.672 2.720 1.584 1.552 Cholesterol (mmol/l) week 1 week 4 5.628 6.516 6.176 7.797 6.352 6.360 5.112 5.733 5.224 6.126 6.916 6.099 7.916 5.445 7.336 7.041 6.514 5.916 6.030 5.733 6.32 6.28 0.90 0.70 7.802 7.875 4.290 3.330 6.562 6.282 5.756 5.577 6.160 7.329 4.612 3.171 4.644 4.689 6.900 6.699 5.918 6.699 6.466 6.255 5.91 5.79 1.12 1.60 6.434 5.550 6.014 5.628 6.514 6.231 5.998 5.760 5.370 4.218 5.482 5.655 5.596 5.211 6.402 5.550 6.932 7.563 3.936 3.459 5.87 5.48 0.84 1.10 3.130 3.981 3.244 3.981 4.114 4.428 3.824 3.852 4.644 4.140 3.808 3.903 3.790 4.401 3.758 4.062 4.934 5.262 3.114 3.879 3.84 4.19 0.61 0.43 3.742 3.537 3.050 3.591 3.678 3.486 3.340 3.564 6.112 3.774 4.452 4.164 2.970 2.439 3.276 4.347 4.016 4.062 4.548 4.401 3.92 3.74 0.94 0.57 3.984 3.303 5.208 3.303 5.112 3.669 4.516 3.330 4.984 3.408 5.016 3.330 3.308 3.642 4.660 3.015 4.048 3.747 4.130 3.930 4.50 3.47 0.62 0.27 3.292 7.014 3.550 9.522 3.082 7.719 3.098 6.726 3.002 5.760 3.162 6.231 3.758 5.994 3.856 5.889 3.678 5.472 3.566 5.682 3.40 6.60 0.31 1.24 week 8 6.633 7.182 5.175 5.631 5.199 7.500 6.498 6.885 5.448 5.562 6.17 0.87 6.543 2.601 6.861 4.059 6.816 2.511 3.627 6.453 7.317 6.087 5.29 1.88 7.317 6.063 5.313 6.954 5.289 5.610 5.109 6.657 7.293 3.945 5.96 1.10 3.375 4.059 4.266 3.558 4.470 3.627 4.356 3.468 5.016 4.287 4.05 0.53 3.216 5.655 5.949 4.470 7.773 7.362 2.238 4.287 3.741 6.291 5.10 1.81 5.382 3.741 5.859 4.833 4.629 5.904 2.784 4.356 5.289 7.158 4.99 1.23 6.339 6.063 7.341 6.840 5.448 5.496 5.199 6.474 5.085 4.515 5.88 0.88 Page A13 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 week 0 1 4 8 week 0 1 4 8 week 0 1 4 8 group 1 vs 2 nn p 10 10 0.631 10 10 0.436 10 10 0.912 10 10 0.436 group 2 vs 4 nn p 10 10 0.247 10 10 <0.001 10 10 0.029 10 10 0.190 group 3 vs 7 nn p 10 10 0.739 10 10 <0.001 10 10 0.023 10 10 0.796 group 1 vs 3 nn p 10 10 0.631 10 10 0.393 10 10 0.035 10 10 0.796 group 2 vs 5 nn p 10 10 0.218 10 10 <0.001 10 10 0.015 10 10 0.796 group 4 vs 5 nn p 10 10 0.739 10 10 0.739 10 10 0.089 10 10 0.165 Plasma cholesterol, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 group 1 vs 7 nn p nn p nn p nn p 10 10 0.631 10 10 0.393 10 10 0.247 10 10 0.912 10 10 <0.001 10 10 <0.001 10 10 <0.001 10 10 <0.001 10 10 <0.001 10 10 <0.001 10 10 <0.001 10 10 0.853 10 10 <0.001 10 10 0.247 10 10 0.035 10 10 0.393 group 2 vs 6 group 2 vs 7 group 3 vs 4 group 3 vs 5 nn p nn p nn p nn p 10 10 0.190 10 10 0.529 10 10 0.353 10 10 0.315 10 10 0.009 10 10 <0.001 10 10 <0.001 10 10 0.001 10 10 0.007 10 10 0.579 10 10 0.005 10 10 0.002 10 10 0.579 10 10 0.853 10 10 <0.001 10 10 0.393 group 4 vs 6 group 4 vs 7 group 5 vs 6 group 5 vs 7 nn p nn p nn p nn p 10 10 0.393 10 10 0.654 10 10 0.631 10 10 0.436 10 10 0.015 10 10 0.052 10 10 0.052 10 10 0.247 10 10 <0.001 10 10 <0.001 10 10 0.089 10 10 <0.001 10 10 0.023 10 10 <0.001 10 10 0.853 10 10 0.315 group 2 vs 3 nn p 10 10 0.796 10 10 0.853 10 10 0.393 10 10 0.579 group 3 vs 6 nn p 10 10 0.123 10 10 0.001 10 10 <0.001 10 10 0.089 group 6 vs 7 nn p 10 10 0.190 10 10 <0.001 10 10 <0.001 10 10 0.089 p<0.05 Page A14 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 C2. Plasma triglyceride measurements group Mouse# cage# 1 1 1 1 2 1 1 3 2 1 4 2 1 5 3 1 6 3 1 7 4 1 8 5 1 9 5 1 10 5 AVERAGE SD 2 11 6 2 12 6 2 13 6 2 14 7 2 15 8 2 16 8 2 17 8 2 18 9 2 19 9 2 20 10 AVERAGE SD 3 21 11 3 22 12 3 23 13 3 24 13 3 25 14 3 26 15 3 27 15 3 28 15 3 29 16 3 30 17 AVERAGE SD 4 31 18 4 32 18 4 33 19 4 34 19 4 35 19 4 36 19 4 37 20 4 38 20 4 39 21 4 40 21 AVERAGE SD 5 41 22 5 42 22 5 43 22 5 44 22 5 45 23 5 46 24 5 47 25 5 48 25 5 49 26 5 50 27 AVERAGE SD 6 51 28 6 52 29 6 53 29 6 54 30 6 55 30 6 56 31 6 57 31 6 58 31 6 59 31 6 60 32 AVERAGE SD 7 61 33 7 62 34 7 63 35 7 64 36 7 65 36 7 66 37 7 67 38 7 68 38 7 69 39 7 70 39 AVERAGE SD excluded 655 85 excluded 656 86 excluded 741 96 excluded 742 96 excluded 748 97 eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 12 00 21 02 20 Measured TG (mmol/l) week 0 week 1 week 4 week 8 0.445 0.387 0.257 0.298 0.467 0.270 0.263 0.266 0.950 0.731 0.290 0.233 0.706 0.609 0.329 0.277 1.666 0.583 0.214 0.277 0.645 0.472 0.345 0.244 0.595 0.625 0.247 0.471 0.495 0.482 0.208 0.293 0.562 0.477 0.296 0.233 0.756 0.456 0.186 0.212 week 0 2 2 2 2 2 2 2 2 2 2 dilution week 1 week 4 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 2 3 week 8 3 3 3 3 3 3 3 3 3 3 0.584 0.434 0.274 0.255 2 2 3 3 0.695 0.667 0.356 0.336 2 2 3 3 0.639 0.445 0.219 0.255 2 2 3 3 0.961 0.535 0.219 0.222 2 2 3 3 1.366 0.842 0.340 0.461 2 2 3 3 0.833 0.688 0.422 0.655 2 2 3 3 0.417 1.080 0.515 0.585 2 2 3 3 0.589 0.376 0.257 0.201 2 2 3 3 0.467 0.477 0.252 0.320 2 2 3 3 0.501 0.355 0.214 0.195 2 2 3 3 0.573 0.276 0.121 0.190 2 2 3 3 0.606 0.307 0.148 0.201 2 2 3 3 0.978 0.762 0.307 0.233 2 2 3 3 0.661 0.932 0.372 0.168 2 2 3 3 0.795 0.456 0.143 0.190 2 2 3 3 0.656 0.482 0.148 0.244 2 2 3 3 0.478 0.376 0.143 0.152 2 2 3 3 0.528 0.382 0.137 0.195 2 2 3 3 0.412 0.403 0.208 0.336 2 2 3 3 0.900 0.673 0.312 0.369 2 2 3 3 0.856 0.244 0.137 0.082 2 2 3 3 0.634 0.281 0.143 0.082 2 2 3 3 0.445 0.276 0.121 0.087 2 2 3 3 0.506 0.233 0.099 0.103 2 2 3 3 0.589 0.207 0.132 0.093 2 2 3 3 0.489 0.297 0.230 0.206 2 2 3 3 0.739 0.329 0.132 0.082 2 2 3 3 0.994 0.419 0.148 0.082 2 2 3 3 0.562 0.270 0.104 0.103 2 2 3 3 0.673 0.339 0.186 0.190 2 2 3 3 0.229 0.376 0.121 0.120 2 2 3 3 0.456 0.212 0.132 0.228 2 2 3 3 0.512 0.165 0.099 0.206 2 2 3 3 1.194 0.202 0.132 0.222 2 2 3 3 0.928 0.218 0.099 0.358 2 2 3 3 0.634 0.181 0.110 0.320 2 2 3 3 0.722 0.260 0.247 0.309 2 2 3 3 0.678 0.207 0.208 0.250 2 2 3 3 0.589 0.323 0.870 0.103 2 2 3 3 0.551 0.186 0.115 0.260 2 2 3 3 0.728 0.424 0.110 0.131 2 2 3 3 0.545 0.773 0.170 0.125 2 2 3 3 0.850 0.620 0.208 0.201 2 2 3 3 1.227 0.916 0.159 0.136 2 2 3 3 0.667 0.720 0.170 0.212 2 2 3 3 0.606 0.487 0.159 0.228 2 2 3 3 0.168 0.244 0.154 0.114 2 2 3 3 0.628 0.588 0.154 0.179 2 2 3 3 0.578 0.419 0.154 0.168 2 2 3 3 0.467 0.366 0.126 0.325 2 2 3 3 1.349 0.196 0.263 0.168 2 2 3 3 0.717 0.228 0.361 0.163 2 2 3 3 0.650 0.249 0.356 0.212 2 2 3 3 0.856 0.244 0.203 0.217 2 2 3 3 0.784 0.265 0.236 0.260 2 2 3 3 0.539 0.329 0.318 0.212 2 2 3 3 0.606 0.202 0.312 0.201 2 2 3 3 0.428 0.265 0.225 0.379 2 2 3 3 0.573 0.302 0.230 0.212 2 2 3 3 0.473 0.255 0.132 0.136 2 2 3 3 1.954 2 0.978 2 0.917 2 0.251 2 0.256 2 week 0 0.89 0.934 1.9 1.412 3.332 1.29 1.19 0.99 1.124 1.512 1.46 0.73 1.168 1.39 1.278 1.922 2.732 1.666 0.834 1.178 0.934 1.002 1.41 0.57 1.146 1.212 1.956 1.322 1.59 1.312 0.956 1.056 0.824 1.8 1.32 0.36 1.712 1.268 0.89 1.012 1.178 0.978 1.478 1.988 1.124 1.346 1.30 0.35 0.458 0.912 1.024 2.388 1.856 1.268 1.444 1.356 1.178 1.102 1.30 0.53 1.456 1.09 1.7 2.454 1.334 1.212 0.336 1.256 1.156 0.934 1.29 0.54 2.698 1.434 1.3 1.712 1.568 1.078 1.212 0.856 1.146 0.946 1.40 0.53 3.908 1.956 1.834 0.502 0.512 TG (mmol/l) week 1 week 4 0.774 0.771 0.54 0.789 1.462 0.87 1.218 0.987 1.166 0.642 0.944 1.035 1.25 0.741 0.964 0.624 0.954 0.888 0.912 0.558 1.02 0.79 0.26 0.16 0.868 0.822 1.334 1.068 0.89 0.657 1.07 0.657 1.684 1.02 1.376 1.266 2.16 1.545 0.752 0.771 0.954 0.756 0.71 0.642 1.18 0.92 0.46 0.30 0.552 0.363 0.614 0.444 1.524 0.921 1.864 1.116 0.912 0.429 0.964 0.444 0.752 0.429 0.764 0.411 0.806 0.624 1.346 0.936 1.01 0.61 0.43 0.27 0.488 0.411 0.562 0.429 0.552 0.363 0.466 0.297 0.414 0.396 0.594 0.69 0.658 0.396 0.838 0.444 0.54 0.312 0.678 0.558 0.58 0.43 0.12 0.12 0.752 0.363 0.424 0.396 0.33 0.297 0.404 0.396 0.436 0.297 0.362 0.33 0.52 0.741 0.414 0.624 0.646 2.61 0.372 0.345 0.47 0.42 0.14 0.16 0.848 0.33 1.546 0.51 1.24 0.624 1.832 0.477 1.44 0.51 0.974 0.477 0.488 0.462 1.176 0.462 0.838 0.462 0.732 0.378 1.11 0.47 0.41 0.08 0.392 0.789 0.456 1.083 0.498 1.068 0.488 0.609 0.53 0.708 0.658 0.954 0.404 0.936 0.53 0.675 0.604 0.69 0.51 0.396 0.51 0.79 0.08 0.22 week 8 0.894 0.798 0.699 0.831 0.831 0.732 1.413 0.879 0.699 0.636 0.84 0.22 0.765 1.008 0.765 0.666 1.383 1.965 1.755 0.603 0.96 0.585 1.05 0.49 0.57 0.603 0.699 0.504 0.57 0.732 0.456 0.585 1.008 1.107 0.68 0.21 0.246 0.246 0.261 0.309 0.279 0.618 0.246 0.246 0.309 0.57 0.33 0.14 0.36 0.684 0.618 0.666 1.074 0.96 0.927 0.75 0.309 0.78 0.76 0.21 0.393 0.375 0.603 0.408 0.636 0.684 0.342 0.537 0.504 0.975 0.55 0.19 0.504 0.489 0.636 0.651 0.78 0.636 0.603 1.137 0.636 0.408 0.65 0.20 Page A15 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 week 0 1 4 8 week 0 1 4 8 week 0 1 4 8 group 1 vs 2 nn p 10 10 0.971 10 10 0.739 10 10 0.393 10 10 0.684 group 2 vs 4 nn p 10 10 0.912 10 10 <0.001 10 10 <0.001 10 10 <0.001 group 3 vs 7 nn p 10 10 1.000 10 10 <0.001 10 10 0.123 10 10 1.000 group 1 vs 3 nn p 10 10 0.971 10 10 0.631 10 10 0.089 10 10 0.043 group 2 vs 5 nn p 10 10 0.796 10 10 <0.001 10 9 <0.001 10 10 0.218 group 4 vs 5 nn p 10 10 1.000 10 10 0.029 10 9 0.400 10 10 <0.001 Plasma triglycerides, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 group 1 vs 7 nn p nn p nn p nn p 10 10 0.912 10 10 0.796 10 10 0.912 10 10 0.912 10 10 <0.001 10 10 <0.001 10 10 0.739 10 10 <0.001 10 10 <0.001 10 9 <0.001 10 10 <0.001 10 10 0.912 10 10 <0.001 10 10 0.353 10 10 0.002 10 10 0.007 group 2 vs 6 group 2 vs 7 group 3 vs 4 group 3 vs 5 nn p nn p nn p nn p 10 10 0.853 10 10 0.971 10 10 0.971 10 10 0.912 10 10 0.853 10 10 <0.001 10 10 0.002 10 10 <0.001 10 10 <0.001 10 10 0.529 10 10 0.063 10 9 0.017 10 10 0.003 10 10 0.029 10 10 0.001 10 10 0.529 group 4 vs 6 group 4 vs 7 group 5 vs 6 group 5 vs 7 nn p nn p nn p nn p 10 10 0.971 10 10 0.853 10 10 0.971 10 10 0.796 10 10 0.001 10 10 0.143 10 10 <0.001 10 10 0.190 10 10 0.143 10 10 0.001 9 10 0.133 9 10 0.001 10 10 0.004 10 10 <0.001 10 10 0.123 10 10 0.315 group 2 vs 3 nn p 10 10 0.971 10 10 0.393 10 10 0.023 10 10 0.029 group 3 vs 6 nn p 10 10 0.971 10 10 0.529 10 10 0.971 10 10 0.123 group 6 vs 7 nn p 10 10 0.796 10 10 0.190 10 10 0.001 10 10 0.315 p<0.05 Page A16 of A172 Appendix to TNO-report of 3M-02 Final version C3. Pooled lipoprotein profile measurements 04-12-2006 Week 0 Cholesterol fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured cholesterol (mM) pooled sample 1 0.008 0.002 0.021 0.043 0.292 0.295 0.166 0.109 0.087 0.109 0.115 0.147 0.222 0.266 0.257 0.216 0.232 0.295 0.307 0.229 0.147 0.093 0.071 0.040 dilution cholesterol (mM) 2 0.016 2 0.004 2 0.042 2 0.086 2 0.584 2 0.590 2 0.332 2 0.218 2 0.174 2 0.218 2 0.230 2 0.294 2 0.444 2 0.532 2 0.514 2 0.432 2 0.464 2 0.590 2 0.614 2 0.458 2 0.294 2 0.186 2 0.142 2 0.080 measured cholesterol (mM) pooled sample 2 0.012 0.008 0.012 0.068 0.248 0.235 0.131 0.093 0.075 0.078 0.100 0.134 0.188 0.222 0.229 0.194 0.210 0.288 0.288 0.203 0.122 0.090 0.049 0.040 dilution cholesterol (mM) 2 0.024 2 0.016 2 0.024 2 0.136 2 0.496 2 0.470 2 0.262 2 0.186 2 0.150 2 0.156 2 0.200 2 0.268 2 0.376 2 0.444 2 0.458 2 0.388 2 0.420 2 0.576 2 0.576 2 0.406 2 0.244 2 0.180 2 0.098 2 0.080 measured cholesterol (mM) pooled sample 3 0.012 0.015 0.005 0.062 0.270 0.285 0.163 0.144 0.103 0.103 0.134 0.181 0.282 0.314 0.279 0.241 0.238 0.339 0.370 0.270 0.166 0.093 0.059 0.040 dilution cholesterol (mM) 2 0.024 2 0.030 2 0.010 2 0.124 2 0.540 2 0.570 2 0.326 2 0.288 2 0.206 2 0.206 2 0.268 2 0.362 2 0.564 2 0.628 2 0.558 2 0.482 2 0.476 2 0.678 2 0.740 2 0.540 2 0.332 2 0.186 2 0.118 2 0.080 Average plasma cholesterol (mmol/l) 0.021 0.017 0.025 0.115 0.540 0.543 0.307 0.231 0.177 0.193 0.233 0.308 0.461 0.535 0.510 0.434 0.453 0.615 0.643 0.468 0.290 0.184 0.119 0.080 sd 0.005 0.013 0.016 0.026 0.044 0.064 0.039 0.052 0.028 0.033 0.034 0.049 0.095 0.092 0.050 0.047 0.029 0.055 0.086 0.068 0.044 0.003 0.022 0.000 PL fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured PL (mg/dl) pooled sample 1 0.000 0.000 0.000 0.000 3.776 4.429 2.305 1.706 1.597 1.652 1.978 3.503 5.083 8.623 7.752 7.261 9.059 12.762 15.104 12.436 10.094 5.954 3.939 1.760 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 PL (mM) 0.000 0.000 0.000 0.000 0.058 0.068 0.036 0.026 0.025 0.026 0.031 0.054 0.079 0.133 0.120 0.112 0.140 0.197 0.234 0.192 0.156 0.092 0.061 0.027 measured PL (mg/dl) pooled sample 2 0.000 0.000 0.003 0.984 3.513 4.235 2.584 2.016 1.706 1.861 2.016 3.409 5.732 7.022 8.416 7.435 8.622 15.229 15.177 13.061 9.913 7.704 3.822 2.171 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 PL (mM) 0.000 0.000 0.000 0.015 0.054 0.065 0.040 0.031 0.026 0.029 0.031 0.053 0.089 0.109 0.130 0.115 0.133 0.235 0.235 0.202 0.153 0.119 0.059 0.034 measured PL (mg/dl) pooled sample 3 0.000 0.000 0.000 0.000 4.145 4.199 2.670 1.850 1.468 1.796 2.561 3.708 7.313 9.389 10.044 8.460 10.099 17.036 19.167 15.944 10.918 7.969 4.418 1.632 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 PL (mM) 0.000 0.000 0.000 0.000 0.064 0.065 0.041 0.029 0.023 0.028 0.040 0.057 0.113 0.145 0.155 0.131 0.156 0.263 0.296 0.246 0.169 0.123 0.068 0.025 Average plasma PL(mM) 0.000 0.000 0.000 0.005 0.059 0.066 0.039 0.029 0.025 0.027 0.034 0.055 0.093 0.129 0.135 0.119 0.143 0.232 0.255 0.214 0.159 0.111 0.063 0.029 sd 0.000 0.000 0.000 0.009 0.005 0.002 0.003 0.002 0.002 0.002 0.005 0.002 0.018 0.019 0.018 0.010 0.012 0.033 0.036 0.029 0.008 0.017 0.005 0.004 Page A17 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 week 4 Cholesterol fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured cholesterol (mM) pool group 1 0.006 0.006 0.006 0.028 0.136 0.139 0.096 0.074 0.071 0.083 0.126 0.197 0.272 0.284 0.241 0.204 0.278 0.442 0.429 0.265 0.151 0.099 0.065 0.040 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 1 0.012 0.012 0.012 0.056 0.272 0.278 0.192 0.148 0.142 0.166 0.252 0.394 0.544 0.568 0.482 0.408 0.556 0.884 0.858 0.530 0.302 0.198 0.130 0.080 measured cholesterol (mM) pool group 2 0.012 0.009 0.009 0.062 0.185 0.157 0.099 0.074 0.071 0.077 0.120 0.173 0.225 0.231 0.219 0.207 0.253 0.358 0.327 0.204 0.123 0.080 0.049 0.034 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 2 0.024 0.018 0.018 0.124 0.370 0.314 0.198 0.148 0.142 0.154 0.240 0.346 0.450 0.462 0.438 0.414 0.506 0.716 0.654 0.408 0.246 0.160 0.098 0.068 measured cholesterol (mM) pool group 3 0.009 0.012 0.009 0.031 0.108 0.142 0.089 0.077 0.068 0.083 0.108 0.157 0.204 0.207 0.182 0.179 0.222 0.337 0.352 0.238 0.136 0.086 0.049 0.031 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 3 0.018 0.024 0.018 0.062 0.216 0.284 0.178 0.154 0.136 0.166 0.216 0.314 0.408 0.414 0.364 0.358 0.444 0.674 0.704 0.476 0.272 0.172 0.098 0.062 fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured cholesterol (mM) pool group 4 0.020 0.014 0.027 0.030 0.064 0.058 0.048 0.030 0.027 0.030 0.054 0.107 0.215 0.324 0.324 0.271 0.231 0.203 0.160 0.107 0.064 0.045 0.036 0.024 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 4 0.040 0.028 0.054 0.060 0.128 0.116 0.096 0.060 0.054 0.060 0.108 0.214 0.430 0.648 0.648 0.542 0.462 0.406 0.320 0.214 0.128 0.090 0.072 0.048 measured cholesterol (mM) pool group 5 0.008 0.008 0.005 0.024 0.076 0.079 0.051 0.036 0.033 0.030 0.030 0.061 0.132 0.209 0.259 0.231 0.209 0.209 0.175 0.116 0.073 0.039 0.036 0.017 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 5 0.016 0.016 0.010 0.048 0.152 0.158 0.102 0.072 0.066 0.060 0.060 0.122 0.264 0.418 0.518 0.462 0.418 0.418 0.350 0.232 0.146 0.078 0.072 0.034 measured cholesterol (mM) pool group 6 0.011 0.008 0.011 0.024 0.054 0.073 0.039 0.036 0.027 0.030 0.036 0.064 0.135 0.215 0.249 0.218 0.197 0.184 0.160 0.110 0.064 0.039 0.027 0.014 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 6 0.022 0.016 0.022 0.048 0.108 0.146 0.078 0.072 0.054 0.060 0.072 0.128 0.270 0.430 0.498 0.436 0.394 0.368 0.320 0.220 0.128 0.078 0.054 0.028 measured cholesterol (mM) pool group 7 0.016 0.013 0.010 0.025 0.120 0.126 0.065 0.049 0.031 0.034 0.043 0.095 0.246 0.436 0.525 0.458 0.359 0.277 0.203 0.129 0.083 0.049 0.040 0.025 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 7 0.032 0.026 0.020 0.050 0.240 0.252 0.130 0.098 0.062 0.068 0.086 0.190 0.492 0.872 1.050 0.916 0.718 0.554 0.406 0.258 0.166 0.098 0.080 0.050 Page A18 of A172 Appendix to TNO-report of 3M-02 Final version PL fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured PL(mg/dl) pool group 1 0.000 0.000 0.000 0.000 0.113 0.247 0.000 0.000 0.000 0.000 0.918 3.201 5.752 7.095 6.289 6.155 12.063 22.804 23.744 14.748 8.303 4.946 2.261 0.247 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 1 0.000 0.000 0.000 0.000 0.002 0.004 0.000 0.000 0.000 0.000 0.014 0.049 0.089 0.110 0.097 0.095 0.186 0.353 0.367 0.228 0.128 0.076 0.035 0.004 measured PL(mg/dl) pool group 2 0.000 0.000 0.000 0.000 1.321 1.187 0.000 0.000 0.000 0.000 0.650 2.664 4.275 5.886 5.618 5.886 10.317 17.702 17.433 10.586 5.752 3.872 1.187 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 2 0.000 0.000 0.000 0.000 0.020 0.018 0.000 0.000 0.000 0.000 0.010 0.041 0.066 0.091 0.087 0.091 0.160 0.274 0.270 0.164 0.089 0.060 0.018 0.000 04-12-2006 measured PL(mg/dl) pool group 3 0.000 0.000 0.000 0.000 0.000 0.516 0.000 0.000 0.000 0.000 0.381 1.992 3.604 4.141 4.409 4.678 8.840 17.031 19.448 12.331 7.363 4.812 1.321 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 3 0.000 0.000 0.000 0.000 0.000 0.008 0.000 0.000 0.000 0.000 0.006 0.031 0.056 0.064 0.068 0.072 0.137 0.263 0.301 0.191 0.114 0.074 0.020 0.000 fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured PL(mg/dl) pool group 4 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.939 4.149 8.162 9.232 8.162 6.557 6.825 5.487 2.946 1.340 0.939 0.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 4 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.015 0.064 0.126 0.143 0.126 0.101 0.106 0.085 0.046 0.021 0.015 0.000 0.000 measured PL(mg/dl) pool group 5 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 1.742 4.551 6.825 6.825 6.022 7.226 6.022 3.481 1.875 0.672 0.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 5 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.027 0.070 0.106 0.106 0.093 0.112 0.093 0.054 0.029 0.010 0.000 0.000 measured PL(mg/dl) pool group 6 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 1.742 4.283 6.022 5.621 5.219 5.487 5.086 2.812 1.207 0.538 0.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 6 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.027 0.066 0.093 0.087 0.081 0.085 0.079 0.043 0.019 0.008 0.000 0.000 measured PL(mg/dl) pool group 7 0.000 0.000 0.000 0.000 0.000 0.187 0.000 0.000 0.000 0.000 0.000 0.858 5.690 11.998 16.024 15.085 12.400 10.253 7.435 4.348 2.603 1.529 0.321 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 7 0.000 0.000 0.000 0.000 0.000 0.003 0.000 0.000 0.000 0.000 0.000 0.013 0.088 0.185 0.248 0.233 0.192 0.159 0.115 0.067 0.040 0.024 0.005 0.000 Page A19 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Week 6 Cholesterol fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured cholesterol (mM) pool group 1 0.010 0.001 0.004 0.025 0.135 0.135 0.083 0.065 0.056 0.074 0.129 0.223 0.293 0.293 0.238 0.183 0.241 0.368 0.347 0.220 0.132 0.092 0.062 0.044 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 1 0.020 0.002 0.008 0.050 0.270 0.270 0.166 0.130 0.112 0.148 0.258 0.446 0.586 0.586 0.476 0.366 0.482 0.736 0.694 0.440 0.264 0.184 0.124 0.088 measured cholesterol (mM) pool group 2 0.007 0.007 0.004 0.050 0.174 0.147 0.083 0.065 0.050 0.068 0.114 0.186 0.226 0.211 0.180 0.162 0.220 0.274 0.241 0.153 0.086 0.059 0.035 0.022 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 2 0.014 0.014 0.008 0.100 0.348 0.294 0.166 0.130 0.100 0.136 0.228 0.372 0.452 0.422 0.360 0.324 0.440 0.548 0.482 0.306 0.172 0.118 0.070 0.044 measured cholesterol (mM) pool group 3 0.013 0.013 0.010 0.062 0.189 0.168 0.092 0.077 0.062 0.083 0.123 0.180 0.223 0.211 0.162 0.156 0.235 0.329 0.296 0.189 0.114 0.074 0.047 0.032 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 3 0.026 0.026 0.020 0.124 0.378 0.336 0.184 0.154 0.124 0.166 0.246 0.360 0.446 0.422 0.324 0.312 0.470 0.658 0.592 0.378 0.228 0.148 0.094 0.064 fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured cholesterol (mM) pool group 4 0.016 0.004 0.010 0.022 0.068 0.056 0.034 0.031 0.025 0.028 0.059 0.138 0.251 0.315 0.300 0.217 0.168 0.129 0.080 0.056 0.037 0.028 0.025 0.019 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 4 0.032 0.008 0.020 0.044 0.136 0.112 0.068 0.062 0.050 0.056 0.118 0.276 0.502 0.630 0.600 0.434 0.336 0.258 0.160 0.112 0.074 0.056 0.050 0.038 measured cholesterol (mM) pool group 5 0.010 0.004 0.010 0.077 0.248 0.193 0.117 0.080 0.071 0.077 0.129 0.220 0.281 0.251 0.202 0.156 0.205 0.281 0.254 0.150 0.092 0.056 0.043 0.037 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 5 0.020 0.008 0.020 0.154 0.496 0.386 0.234 0.160 0.142 0.154 0.258 0.440 0.562 0.502 0.404 0.312 0.410 0.562 0.508 0.300 0.184 0.112 0.086 0.074 measured cholesterol (mM) pool group 6 0.016 0.025 0.016 0.037 0.071 0.071 0.037 0.034 0.031 0.037 0.053 0.120 0.214 0.266 0.266 0.211 0.156 0.126 0.080 0.049 0.037 0.025 0.016 0.013 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 6 0.032 0.050 0.032 0.074 0.142 0.142 0.074 0.068 0.062 0.074 0.106 0.240 0.428 0.532 0.532 0.422 0.312 0.252 0.160 0.098 0.074 0.050 0.032 0.026 measured cholesterol (mM) pool group 7 0.008 0.011 0.011 0.029 0.093 0.081 0.051 0.045 0.036 0.045 0.075 0.142 0.306 0.425 0.407 0.306 0.206 0.139 0.081 0.054 0.039 0.033 0.026 0.017 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 7 0.016 0.022 0.022 0.058 0.186 0.162 0.102 0.090 0.072 0.090 0.150 0.284 0.612 0.850 0.814 0.612 0.412 0.278 0.162 0.108 0.078 0.066 0.052 0.034 Page A20 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 PL fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured PL(mg/dl) pool group 1 0.000 0.000 0.000 0.000 1.035 1.166 0.246 0.115 0.115 1.035 2.349 4.977 6.948 7.605 6.685 6.685 11.415 19.168 18.248 11.678 7.342 4.977 2.086 0.772 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 1 0.000 0.000 0.000 0.000 0.016 0.018 0.004 0.002 0.002 0.016 0.036 0.077 0.107 0.118 0.103 0.103 0.176 0.296 0.282 0.181 0.114 0.077 0.032 0.012 measured PL(mg/dl) pool group 2 0.000 0.000 0.000 0.000 1.823 1.297 0.903 0.640 0.246 0.509 1.823 4.057 5.502 6.159 5.897 6.028 9.313 13.649 12.729 8.525 4.714 2.874 1.035 0.115 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 2 0.000 0.000 0.000 0.000 0.028 0.020 0.014 0.010 0.004 0.008 0.028 0.063 0.085 0.095 0.091 0.093 0.144 0.211 0.197 0.132 0.073 0.044 0.016 0.002 measured PL(mg/dl) pool group 3 0.000 0.000 0.000 0.000 1.429 1.429 0.115 0.115 0.000 0.640 2.743 3.400 4.451 4.845 4.188 5.108 9.050 14.963 14.306 9.182 5.239 3.268 1.035 0.246 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 3 0.000 0.000 0.000 0.000 0.022 0.022 0.002 0.002 0.000 0.010 0.042 0.053 0.069 0.075 0.065 0.079 0.140 0.231 0.221 0.142 0.081 0.051 0.016 0.004 fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured PL(mg/dl) pool group 4 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.505 2.599 6.264 9.405 9.798 7.835 5.872 4.301 2.861 1.683 1.291 0.636 0.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 4 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.008 0.040 0.097 0.145 0.151 0.121 0.091 0.066 0.044 0.026 0.020 0.010 0.000 0.000 measured PL(mg/dl) pool group 5 0.000 0.000 0.000 0.113 2.338 1.945 0.898 0.505 0.505 0.898 2.730 5.479 7.442 7.442 6.133 6.002 8.489 12.808 12.547 8.227 5.217 2.992 1.160 0.243 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 5 0.000 0.000 0.000 0.002 0.036 0.030 0.014 0.008 0.008 0.014 0.042 0.085 0.115 0.115 0.095 0.093 0.131 0.198 0.194 0.127 0.081 0.046 0.018 0.004 measured PL(mg/dl) pool group 6 0.000 0.000 0.000 0.000 0.000 0.505 0.000 0.000 0.000 0.000 0.374 2.338 5.348 7.966 8.882 7.049 5.610 4.039 2.599 1.552 0.898 0.505 0.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 6 0.000 0.000 0.000 0.000 0.000 0.008 0.000 0.000 0.000 0.000 0.006 0.036 0.083 0.123 0.137 0.109 0.087 0.062 0.040 0.024 0.014 0.008 0.000 0.000 measured PL(mg/dl) pool group 7 0.000 0.000 0.000 0.000 0.282 0.414 0.000 0.000 0.000 0.017 0.678 3.583 8.866 13.621 14.413 11.508 7.281 4.640 2.131 1.074 0.678 0.414 0.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 7 0.000 0.000 0.000 0.000 0.004 0.006 0.000 0.000 0.000 0.000 0.010 0.055 0.137 0.211 0.223 0.178 0.113 0.072 0.033 0.017 0.010 0.006 0.000 0.000 Page A21 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Week 8 Cholesterol fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured cholesterol (mM) pool group 1 0.000 0.000 0.000 0.036 0.191 0.167 0.103 0.078 0.078 0.100 0.164 0.267 0.328 0.292 0.240 0.191 0.249 0.346 0.346 0.200 0.109 0.066 0.048 0.027 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 1 0.000 0.000 0.000 0.072 0.382 0.334 0.206 0.156 0.156 0.200 0.328 0.534 0.656 0.584 0.480 0.382 0.498 0.692 0.692 0.400 0.218 0.132 0.096 0.054 measured cholesterol (mM) pool group 2 0.000 0.000 0.002 0.066 0.228 0.182 0.109 0.078 0.069 0.087 0.139 0.200 0.249 0.252 0.237 0.197 0.237 0.276 0.240 0.136 0.078 0.051 0.023 0.020 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 2 0.000 0.000 0.004 0.132 0.456 0.364 0.218 0.156 0.138 0.174 0.278 0.400 0.498 0.504 0.474 0.394 0.474 0.552 0.480 0.272 0.156 0.102 0.046 0.040 measured cholesterol (mM) pool group 3 0.000 0.011 0.011 0.054 0.206 0.179 0.112 0.091 0.075 0.112 0.164 0.246 0.298 0.273 0.237 0.188 0.231 0.310 0.288 0.170 0.097 0.063 0.030 0.020 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 3 0.000 0.022 0.022 0.108 0.412 0.358 0.224 0.182 0.150 0.224 0.328 0.492 0.596 0.546 0.474 0.376 0.462 0.620 0.576 0.340 0.194 0.126 0.060 0.040 fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured cholesterol (mM) pool group 4 0.015 0.000 0.006 0.021 0.061 0.052 0.023 0.021 0.012 0.035 0.044 0.118 0.271 0.357 0.363 0.280 0.205 0.162 0.107 0.067 0.044 0.023 0.018 0.015 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 4 0.030 0.001 0.012 0.042 0.122 0.104 0.046 0.042 0.024 0.070 0.088 0.236 0.542 0.714 0.726 0.560 0.410 0.324 0.214 0.134 0.088 0.046 0.036 0.030 measured cholesterol (mM) pool group 5 0.000 0.000 0.006 0.041 0.153 0.130 0.078 0.058 0.049 0.058 0.098 0.159 0.210 0.225 0.208 0.179 0.199 0.257 0.222 0.144 0.075 0.052 0.029 0.029 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 5 0.000 0.000 0.012 0.082 0.306 0.260 0.156 0.116 0.098 0.116 0.196 0.318 0.420 0.450 0.416 0.358 0.398 0.514 0.444 0.288 0.150 0.104 0.058 0.058 measured cholesterol (mM) pool group 6 0.000 0.006 0.009 0.035 0.107 0.095 0.061 0.044 0.038 0.032 0.067 0.150 0.265 0.372 0.377 0.300 0.239 0.196 0.150 0.092 0.052 0.032 0.021 0.021 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 6 0.001 0.012 0.018 0.070 0.214 0.190 0.122 0.088 0.076 0.064 0.134 0.300 0.530 0.744 0.754 0.600 0.478 0.392 0.300 0.184 0.104 0.064 0.042 0.042 measured cholesterol (mM) pool group 7 0.000 0.000 0.003 0.041 0.167 0.135 0.073 0.056 0.038 0.050 0.079 0.170 0.302 0.419 0.419 0.328 0.246 0.199 0.138 0.085 0.053 0.035 0.023 0.011 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average cholesterol (mM) group 7 0.000 0.000 0.006 0.082 0.334 0.270 0.146 0.112 0.076 0.100 0.158 0.340 0.604 0.838 0.838 0.656 0.492 0.398 0.276 0.170 0.106 0.070 0.046 0.022 Page A22 of A172 Appendix to TNO-report of 3M-02 Final version PL fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured PL(mg/dl) pool group 1 0.000 0.000 0.000 0.000 2.067 1.800 0.601 0.201 0.334 1.400 3.266 6.599 8.998 8.732 7.132 7.265 11.931 20.595 18.596 11.531 7.265 4.733 2.467 0.867 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 1 0.000 0.000 0.000 0.000 0.032 0.028 0.009 0.003 0.005 0.022 0.050 0.102 0.139 0.135 0.110 0.112 0.184 0.318 0.287 0.178 0.112 0.073 0.038 0.013 measured PL(mg/dl) pool group 2 0.000 0.000 0.000 0.201 3.000 2.067 0.867 0.201 0.334 1.000 2.600 4.999 6.866 7.932 7.665 8.065 10.731 15.130 12.864 8.065 5.266 3.266 1.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 2 0.000 0.000 0.000 0.003 0.046 0.032 0.013 0.003 0.005 0.015 0.040 0.077 0.106 0.123 0.119 0.125 0.166 0.234 0.199 0.125 0.081 0.050 0.015 0.000 04-12-2006 measured PL(mg/dl) pool group 3 0.000 0.000 0.000 0.000 2.200 2.067 0.867 0.334 0.734 1.400 3.533 6.199 8.199 8.732 7.265 7.399 10.864 17.929 15.930 9.798 6.732 4.200 1.534 0.334 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 3 0.000 0.000 0.000 0.000 0.034 0.032 0.013 0.005 0.011 0.022 0.055 0.096 0.127 0.135 0.112 0.114 0.168 0.277 0.246 0.151 0.104 0.065 0.024 0.005 fraction 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 measured PL(mg/dl) pool group 4 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.048 2.583 8.185 12.587 12.987 10.453 8.052 6.184 4.183 2.583 1.916 1.115 0.000 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 4 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.000 0.001 0.040 0.127 0.195 0.201 0.162 0.124 0.096 0.065 0.040 0.030 0.017 0.000 0.000 measured PL(mg/dl) pool group 5 0.000 0.000 0.000 0.000 1.515 1.249 0.181 0.000 0.000 0.448 1.515 3.916 5.517 6.851 6.851 7.118 9.786 14.454 13.521 8.318 4.450 3.116 0.848 0.315 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 5 0.000 0.000 0.000 0.000 0.023 0.019 0.003 0.000 0.000 0.007 0.023 0.061 0.085 0.106 0.106 0.110 0.151 0.223 0.209 0.129 0.069 0.048 0.013 0.005 measured PL(mg/dl) pool group 6 0.000 0.000 0.000 0.000 0.715 0.582 0.000 0.000 0.000 0.000 0.715 3.783 8.452 12.720 13.120 11.386 9.519 8.318 5.917 3.650 2.983 2.182 0.315 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 6 0.000 0.000 0.000 0.000 0.011 0.009 0.000 0.000 0.000 0.000 0.011 0.058 0.131 0.197 0.203 0.176 0.147 0.129 0.091 0.056 0.046 0.034 0.005 0.000 measured PL(mg/dl) pool group 7 0.000 0.000 0.000 0.000 1.619 0.951 0.150 0.000 0.000 0.000 0.951 4.290 10.033 13.639 14.707 12.570 10.166 8.163 5.626 3.622 2.687 1.619 0.150 0.000 dilution 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Average PL (mM) group 7 0.000 0.000 0.000 0.000 0.025 0.015 0.002 0.000 0.000 0.000 0.015 0.066 0.155 0.211 0.227 0.194 0.157 0.126 0.087 0.056 0.042 0.025 0.002 0.000 Page A23 of A172 Appendix to TNO-report of 3M-02 Final version C4. Pooled plasma ALAT measurements 04-12-2006 group Group 1: HF control Group 2: HF + 0.01 % PFBS Group 3: HF + 0.03 % PFBS Group 4: HF + 0.006 % PFHS Group 5: HF + 0.02 % PFHS Group 6: HF + 0.003 % PFOS Group 7: HF + 0.01 % PFOS week 0 97.1 measured week 4 27.1 30.6 23.5 88.7 46 109 119 Pooled plasma ALAT (U/l) dilution week 4 week 6 (o/n fast) 3 81.3 84 3 91.8 113 3 70.5 108 3 266.1 213 3 138 130 3 327 246 3 357 307 week 8 57 134 87.2 370 180 394 450 Page A24 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 C5. Plasma ketone bodies measurements Week -1 concentration ketonbodies (mmol/l) 0.000 0.000 0.033 0.033 0.067 0.067 0.134 0.134 0.268 0.268 0.535 0.535 1.070 1.070 2.140 2.140 A340 pre 0.214 0.214 0.218 0.215 0.218 0.222 0.216 0.22 0.218 0.215 0.218 0.215 0.217 0.218 0.217 0.22 A340 post 0.211 0.214 0.222 0.22 0.223 0.239 0.249 0.257 0.282 0.284 0.348 0.358 0.462 0.484 0.689 0.716 A340 netto -0.003 0 0.004 0.005 0.005 0.017 0.033 0.037 0.064 0.069 0.13 0.143 0.245 0.266 0.472 0.496 A340netto 0.6 y = 0.2283x + 0.0028 0.5 0.4 0.3 0.2 0.1 0 0.0 0.5 1.0 1.5 2.0 2.5 Keton bodies (mmol/l) y=ax + b mouse# 641 645 646 650 652 655 656 657 658 663 665 666 667 670 671 674 678 679 680 681 686 687 690 696 699 700 704 705 706 710 712 714 716 720 721 724 727 729 730 731 737 738 740 741 742 744 748 752 754 756 758 759 760 761 762 763 765 766 768 769 771 774 776 778 782 783 784 785 790 791 792 793 795 797 799 a: A340 pre 0.307 0.321 0.3 0.269 0.293 0.329 0.307 0.381 0.293 0.35 0.245 0.276 0.273 0.351 0.324 0.464 0.291 0.447 0.334 0.468 0.352 0.467 0.356 0.274 0.3 0.305 0.311 0.294 0.292 0.419 0.388 0.289 0.333 0.346 0.302 0.347 0.29 0.244 0.317 0.27 0.432 0.387 0.351 0.249 0.239 0.242 0.26 0.284 0.366 0.282 0.333 0.317 0.303 0.293 0.277 0.297 0.287 0.309 0.383 0.382 0.318 0.357 0.31 0.241 0.298 0.282 0.284 0.272 0.253 0.326 0.292 0.261 0.371 0.339 0.301 0.2283 A340 post 0.766 0.546 0.623 0.644 0.546 0.53 0.505 0.67 0.658 0.58 0.489 0.65 0.668 0.621 0.613 0.628 0.534 0.649 0.579 0.736 0.608 0.638 0.644 0.546 0.563 0.583 0.666 0.609 0.621 0.664 0.624 0.573 0.465 0.692 0.705 0.754 0.44 0.37 0.423 0.405 0.712 0.581 0.578 0.485 0.418 0.516 0.429 0.445 0.631 0.558 0.664 0.613 0.605 0.639 0.623 0.69 0.62 0.627 0.632 0.693 0.533 0.554 0.521 0.466 0.596 0.556 0.653 0.645 0.452 0.589 0.58 0.403 0.561 0.712 0.572 b: 0.0028 A340 netto 0.459 0.225 0.323 0.375 0.253 0.201 0.198 0.289 0.365 0.23 0.244 0.374 0.395 0.27 0.289 0.164 0.243 0.202 0.245 0.268 0.256 0.171 0.288 0.272 0.263 0.278 0.355 0.315 0.329 0.245 0.236 0.284 0.132 0.346 0.403 0.407 0.15 0.126 0.106 0.135 0.28 0.194 0.227 0.236 0.179 0.274 0.169 0.161 0.265 0.276 0.331 0.296 0.302 0.346 0.346 0.393 0.333 0.318 0.249 0.311 0.215 0.197 0.211 0.225 0.298 0.274 0.369 0.373 0.199 0.263 0.288 0.142 0.19 0.373 0.271 meas. concentration ketonbodies (mmol/l) 2.00 0.97 1.40 1.63 1.10 0.87 0.86 1.25 1.59 1.00 1.06 1.63 1.72 1.17 1.25 0.71 1.05 0.87 1.06 1.16 1.11 0.74 1.25 1.18 1.14 1.21 1.54 1.37 1.43 1.06 1.02 1.23 0.57 1.50 1.75 1.77 0.64 0.54 0.45 0.58 1.21 0.84 0.98 1.02 0.77 1.19 0.73 0.69 1.15 1.20 1.44 1.28 1.31 1.50 1.50 1.71 1.45 1.38 1.08 1.35 0.93 0.85 0.91 0.97 1.29 1.19 1.60 1.62 0.86 1.14 1.25 0.61 0.82 1.62 1.17 dilution 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 2 2 2 1 1 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 1 1 1 Plasma keton- bodies (mmol/l) 2.00 0.97 1.40 1.63 1.10 0.87 0.86 1.25 1.59 1.00 1.06 1.63 1.72 1.17 1.25 0.71 1.05 0.87 1.06 1.16 1.11 0.74 1.25 1.18 1.14 1.21 1.54 1.37 1.43 1.06 1.02 1.23 0.57 1.50 1.75 1.77 1.29 1.08 0.90 1.16 1.21 0.84 0.98 1.02 0.77 1.19 0.73 1.39 1.15 1.20 1.44 1.28 1.31 1.50 1.50 1.71 1.45 1.38 1.08 1.35 0.93 0.85 0.91 0.97 1.29 1.19 1.60 1.62 1.72 1.14 1.25 0.61 0.82 1.62 1.17 mouse# after matching 11 12 13 21 61 excluded excluded 62 1 2 41 42 43 63 44 14 22 45 64 31 32 65 23 66 24 46 33 34 35 36 3 51 4 67 52 68 53 25 47 48 15 16 17 excluded excluded 49 excluded 5 26 27 6 28 18 69 70 7 19 29 37 54 38 55 56 57 58 59 8 20 9 10 39 30 40 60 50 Page A25 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Week 6 concentration ketonbodies (mmol/l) 0.000 0.000 0.033 0.033 0.067 0.067 0.134 0.134 0.268 0.268 0.535 0.535 1.070 1.070 2.140 2.140 A340 pre 0.255 0.254 0.255 0.258 0.256 0.262 0.257 0.262 0.257 0.256 0.258 0.257 0.256 0.261 0.259 0.266 A340 post 0.242 0.246 0.251 0.254 0.261 0.269 0.279 0.287 0.318 0.319 0.388 0.392 0.5 0.521 0.729 0.768 A340 netto -0.013 -0.008 -0.004 -0.004 0.005 0.007 0.022 0.025 0.061 0.063 0.13 0.135 0.244 0.26 0.47 0.502 A340netto 0.6 y = 0.233x - 0.0053 0.5 0.4 0.3 0.2 0.1 0 0.0 0.5 1.0 1.5 2.0 2.5 Keton bodies (mmol/l) y=ax + b Group 1 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 7 AVERAGE SD a: 0.233 Mouse# cage# 1 1 2 1 3 2 4 2 5 3 6 3 7 4 8 5 9 5 10 5 11 6 12 6 13 6 14 7 15 8 16 8 17 8 18 9 19 9 20 10 21 11 22 12 23 13 24 13 25 14 26 15 27 15 28 15 29 16 30 17 31 18 32 18 33 19 34 19 35 19 36 19 37 20 38 20 39 21 40 21 41 22 42 22 43 22 44 22 45 23 46 24 47 25 48 25 49 26 50 27 51 28 52 29 53 29 54 30 55 30 56 31 57 31 58 31 59 31 60 32 61 33 62 34 63 35 64 36 65 36 66 37 67 38 68 38 69 39 70 39 b: -0.0053 eartag L R 0 1 1 2 0 0 2 0 0 0 0 2 1 1 0 0 0 2 1 2 A340 pre 0.303 0.284 0.273 0.271 0.267 0.27 0.283 0.27 0.269 0.28 1 0 0.289 2 1 0.288 0 2 0.273 0 1 0.288 1 0 0.282 0 1 0.299 2 0 0.308 0 0 0.271 2 1 0.267 1 0 0.27 0 1 0.279 0 2 0.277 0 1 0.318 1 1 0.291 1 0 0.262 0 1 0.272 2 0 0.261 1 2 0.269 0 2 0.28 1 0 0.269 1 0 0.262 0 2 0.266 0 0 0.275 1 0 0.275 0 1 0.285 0 2 0.284 0 0 0.263 1 1 0.263 0 0 0.272 1 1 0.273 1 0 0.259 0 1 0.29 1 1 0.299 1 2 0.284 1 2 0.315 2 0 0.305 0 1 0.309 1 1 0.295 0 0 0.253 1 2 0.296 0 1 0.263 1 0 0.261 1 2 0.276 1 0 0.262 0 2 0.262 0 0 0.278 0 1 0.264 0 2 0.274 2 2 0.283 2 1 0.284 2 0 0.266 1 0 0.269 0 2 0.268 0 0 0.268 1 2 0.268 0 0 0.27 0 0 0.268 2 0 0.269 1 0 0.277 0 1 0.274 A340 post 0.443 0.402 0.383 0.359 0.382 0.363 0.424 0.38 0.367 0.379 0.413 0.343 0.382 0.468 0.395 0.347 0.364 0.413 0.366 0.383 0.443 0.473 0.409 0.38 0.421 0.418 0.419 0.374 0.445 0.372 0.409 0.381 0.424 0.439 0.452 0.446 0.417 0.365 0.49 0.427 0.435 0.437 0.541 0.408 0.558 0.529 0.399 0.439 0.44 0.466 0.581 0.556 0.541 0.54 0.551 0.605 0.408 0.605 0.564 0.659 0.551 0.568 0.566 0.522 0.541 0.579 0.615 0.568 0.618 0.647 A340 netto 0.14 0.118 0.11 0.088 0.115 0.093 0.141 0.11 0.098 0.099 0.124 0.055 0.109 0.18 0.113 0.048 0.056 0.142 0.099 0.113 0.164 0.196 0.091 0.089 0.159 0.146 0.158 0.105 0.165 0.103 0.147 0.115 0.149 0.164 0.167 0.162 0.154 0.102 0.218 0.154 0.176 0.147 0.242 0.124 0.243 0.224 0.09 0.144 0.187 0.17 0.318 0.295 0.265 0.278 0.289 0.327 0.144 0.331 0.281 0.375 0.285 0.299 0.298 0.254 0.273 0.309 0.347 0.299 0.341 0.373 meas. concentration ketonbodies (mmol/l) 0.62 0.53 0.49 0.40 0.52 0.42 0.63 0.49 0.44 0.45 0.55 0.26 0.49 0.80 0.51 0.23 0.26 0.63 0.45 0.51 0.73 0.86 0.41 0.40 0.71 0.65 0.70 0.47 0.73 0.46 0.65 0.52 0.66 0.73 0.74 0.72 0.68 0.46 0.96 0.68 0.78 0.65 1.06 0.55 1.07 0.98 0.41 0.64 0.83 0.75 1.39 1.29 1.16 1.22 1.26 1.43 0.64 1.44 1.23 1.63 1.25 1.31 1.30 1.11 1.19 1.35 1.51 1.31 1.49 1.62 dilution 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 Plasma keton- bodies (mmol/l) 1.87 1.59 1.48 1.20 1.55 1.27 1.88 1.48 1.33 1.34 1.50 0.23 1.66 0.78 1.47 2.39 1.52 0.69 0.79 1.90 1.34 1.52 1.41 0.54 2.18 2.59 1.24 1.21 2.12 1.95 2.10 1.42 2.19 1.39 1.84 0.48 1.96 1.55 1.99 2.18 2.22 2.15 2.05 1.38 2.88 2.05 2.04 0.40 2.33 1.96 3.18 1.66 3.20 2.95 1.23 1.92 2.48 2.26 2.32 0.65 4.16 3.87 3.48 3.65 3.79 4.28 1.92 4.33 3.69 4.90 3.81 0.78 3.74 3.92 3.91 3.34 3.58 4.05 4.54 3.92 4.46 4.87 4.03 0.47 Page A26 of A172 Appendix to TNO-report of 3M-02 Final version Group Mouse# cage# 1 1 1 1 2 1 1 3 2 1 4 2 1 5 3 1 6 3 1 7 4 1 8 5 1 9 5 1 10 5 AVERAGE SD 2 11 6 2 12 6 2 13 6 2 14 7 2 15 8 2 16 8 2 17 8 2 18 9 2 19 9 2 20 10 AVERAGE SD 3 21 11 3 22 12 3 23 13 3 24 13 3 25 14 3 26 15 3 27 15 3 28 15 3 29 16 3 30 17 AVERAGE SD 4 31 18 4 32 18 4 33 19 4 34 19 4 35 19 4 36 19 4 37 20 4 38 20 4 39 21 4 40 21 AVERAGE SD 5 41 22 5 42 22 5 43 22 5 44 22 5 45 23 5 46 24 5 47 25 5 48 25 5 49 26 5 50 27 AVERAGE SD 6 51 28 6 52 29 6 53 29 6 54 30 6 55 30 6 56 31 6 57 31 6 58 31 6 59 31 6 60 32 AVERAGE SD 7 61 33 7 62 34 7 63 35 7 64 36 7 65 36 7 66 37 7 67 38 7 68 38 7 69 39 7 70 39 AVERAGE SD excluded (655) excluded (656) excluded (741) excluded (742) excluded (748) eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 Plasma ketonbodies (mmol/l) week -1 week 6 1.59 1.87 1.00 1.59 1.02 1.48 0.57 1.20 1.39 1.55 1.44 1.27 1.71 1.88 1.60 1.48 1.72 1.33 1.14 1.34 1.32 1.50 0.38 0.23 2.00 1.66 0.97 0.78 1.40 1.47 0.71 2.39 1.21 1.52 0.84 0.69 0.98 0.79 1.31 1.90 1.45 1.34 1.62 1.52 1.25 1.41 0.39 0.54 1.63 2.18 1.05 2.59 1.25 1.24 1.14 1.21 1.08 2.12 1.15 1.95 1.20 2.10 1.28 1.42 1.38 2.19 0.61 1.39 1.18 1.84 0.26 0.48 1.16 1.96 1.11 1.55 1.54 1.99 1.37 2.18 1.43 2.22 1.06 2.15 1.08 2.05 0.93 1.38 1.25 2.88 0.82 2.05 1.17 2.04 0.22 0.40 1.06 2.33 1.63 1.96 1.72 3.18 1.25 1.66 0.87 3.20 1.21 2.95 0.90 1.23 1.16 1.92 1.19 2.48 1.17 2.26 1.22 2.32 0.27 0.65 1.23 4.16 1.75 3.87 1.29 3.48 1.35 3.65 0.85 3.79 0.91 4.28 0.97 1.92 1.29 4.33 1.19 3.69 1.62 4.90 1.25 3.81 0.29 0.78 1.10 3.74 1.25 3.92 1.17 3.91 1.06 3.34 0.74 3.58 1.18 4.05 1.50 4.54 1.77 3.92 1.50 4.46 1.50 4.87 1.28 4.03 0.30 0.47 0.87 0.86 1.02 0.77 0.73 04-12-2006 Page A27 of A172 Appendix to TNO-report of 3M-02 Final version Appendix D. Measurements in metabolic cages D1. Code mice and metabolic cages group# mouse# metabolic cage# 1 2 101 1 3 105 1 4 201 1 6 205 3 21 102 3 22 106 3 24 202 3 27 206 4 31 103 4 32 107 4 37 203 4 38 207 6 51 104 6 53 108 6 54 204 6 55 208 04-12-2006 D2. Average bodyweight mice in metabolic cages group# mouse# 1 2 1 3 1 4 1 6 3 21 3 22 3 24 3 27 4 31 4 32 4 37 4 38 6 51 6 53 6 54 6 55 metabolic cage# 101 105 201 205 102 106 202 206 103 107 203 207 104 108 204 208 Bodyweight (g) begin metabolic cage end metabolic cage 29.9 30.7 30.2 30 28 29.1 29.3 29.6 29.5 30.2 29.8 30.1 28.8 29.7 29.5 30.2 28.9 29.8 28.9 29 26.9 28.9 30.4 31.3 28.2 29.1 27.3 26.5 29.4 30.7 27.8 28.8 Average bodyweight (g) in metabolic cage 30.30 30.10 28.55 29.45 29.85 29.95 29.25 29.85 29.35 28.95 27.90 30.85 28.65 26.90 30.05 28.30 Page A28 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 D3. Food and drink intake measurements Food intake measurements Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 1: HF control Accumulated food intake (g) Average acc. SD cage 0101 cage 0105 cage 0201 cage 0205 food intake (g) (g) 0.258 0.182 0.114 0.206 0.19 0.06 0.735 0.468 0.273 0.555 0.51 0.19 0.888 0.696 0.733 0.777 0.77 0.08 1.017 0.851 0.926 0.997 0.95 0.08 1.205 0.946 1.021 1.184 1.09 0.13 1.380 1.248 1.356 1.345 1.33 0.06 1.571 1.388 1.770 1.525 1.56 0.16 1.810 1.510 2.016 1.681 1.75 0.21 2.015 1.682 2.182 1.792 1.92 0.22 2.184 1.747 2.294 1.898 2.03 0.25 2.402 1.837 2.324 1.992 2.14 0.27 2.458 1.940 2.623 2.060 2.27 0.32 2.616 2.045 3.224 2.152 2.51 0.54 2.724 2.188 3.390 2.308 2.65 0.54 2.824 2.437 3.480 2.473 2.80 0.48 3.064 2.599 3.486 2.689 2.96 0.40 3.300 2.792 3.510 2.748 3.09 0.38 3.564 3.013 3.525 2.995 3.27 0.31 3.879 3.109 3.676 3.304 3.49 0.35 4.146 3.214 3.802 3.448 3.65 0.41 4.399 3.284 3.980 3.750 3.85 0.46 4.531 3.372 4.110 3.918 3.98 0.48 4.743 3.412 4.174 3.977 4.08 0.55 4.856 3.514 4.280 4.085 4.18 0.55 4.935 3.602 4.321 4.343 4.30 0.55 5.129 3.739 4.441 4.488 4.45 0.57 5.509 4.064 4.469 4.635 4.67 0.61 5.770 4.341 4.705 4.815 4.91 0.61 5.968 4.531 4.897 4.947 5.09 0.62 6.096 4.684 5.032 5.059 5.22 0.61 6.260 4.869 5.144 5.138 5.35 0.62 6.378 5.016 5.351 5.318 5.52 0.59 6.529 5.181 5.535 5.470 5.68 0.59 6.741 5.397 5.692 5.729 5.89 0.59 6.916 5.608 5.944 5.965 6.11 0.56 7.046 5.794 6.060 6.139 6.26 0.54 7.233 5.993 6.328 6.449 6.50 0.52 7.443 6.101 6.615 6.749 6.73 0.55 7.561 6.231 6.776 6.879 6.86 0.55 7.629 6.317 7.002 7.007 6.99 0.54 7.763 6.406 7.175 7.153 7.12 0.56 8.003 6.555 7.392 7.280 7.31 0.59 8.140 6.690 7.504 7.345 7.42 0.60 8.297 6.796 7.656 7.432 7.55 0.62 8.485 6.840 7.831 7.553 7.68 0.68 8.671 7.034 7.922 7.620 7.81 0.68 8.766 7.102 7.973 7.671 7.88 0.69 8.926 7.156 8.014 7.735 7.96 0.74 9.030 7.190 8.075 7.826 8.03 0.76 9.096 7.277 8.149 7.856 8.09 0.76 9.128 7.336 8.299 7.904 8.17 0.75 9.139 7.553 8.481 8.007 8.30 0.68 9.279 7.633 8.613 8.157 8.42 0.70 9.441 7.933 8.781 8.277 8.61 0.66 9.564 8.168 8.938 8.488 8.79 0.61 9.710 8.462 9.207 8.706 9.02 0.55 9.835 8.621 9.459 8.970 9.22 0.53 10.044 8.689 9.575 9.184 9.37 0.58 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 3: HF + 0.03 % PFBS Accumulated food intake (g) Average acc. SD cage 0102 cage 0106 cage 0202 cage 0206 food intake (g) (g) 0.215 0.190 0.188 0.203 0.20 0.01 0.548 0.528 0.643 0.494 0.55 0.06 0.834 0.665 0.961 0.615 0.77 0.16 0.979 0.822 1.200 0.745 0.94 0.20 1.156 1.081 1.312 0.890 1.11 0.18 1.416 1.340 1.505 1.058 1.33 0.19 1.624 1.437 1.738 1.282 1.52 0.20 1.828 1.652 1.911 1.374 1.69 0.24 2.005 1.696 2.105 1.503 1.83 0.28 2.164 1.772 2.282 1.601 1.95 0.32 2.200 1.844 2.450 1.658 2.04 0.35 2.290 1.877 2.731 1.761 2.16 0.44 2.416 2.008 2.955 1.807 2.30 0.51 2.506 2.171 3.097 1.885 2.41 0.52 2.638 2.304 3.284 2.120 2.59 0.51 2.790 2.564 3.633 2.294 2.82 0.58 3.033 2.637 3.819 2.454 2.99 0.61 3.288 2.836 4.141 2.726 3.25 0.64 3.478 3.028 4.557 2.931 3.50 0.74 3.658 3.030 4.845 3.087 3.66 0.84 3.815 3.033 4.958 3.251 3.76 0.86 3.989 3.083 5.184 3.466 3.93 0.91 4.160 3.140 5.331 3.592 4.06 0.95 4.264 3.200 5.478 3.741 4.17 0.97 4.385 3.322 5.596 3.824 4.28 0.98 4.660 3.464 5.823 4.060 4.50 1.01 5.208 3.675 6.138 4.316 4.83 1.07 5.472 3.794 6.381 4.484 5.03 1.13 5.665 4.016 6.645 4.648 5.24 1.16 5.794 4.175 6.781 4.691 5.36 1.16 5.890 4.375 6.887 4.736 5.47 1.14 6.059 4.476 7.033 4.870 5.61 1.16 6.241 4.642 7.204 4.965 5.76 1.18 6.399 4.839 7.392 5.180 5.95 1.17 6.520 5.095 7.600 5.323 6.13 1.16 6.764 5.403 7.862 5.512 6.39 1.16 7.101 5.597 8.127 5.755 6.65 1.20 7.424 5.802 8.354 6.043 6.91 1.20 7.608 5.958 8.528 6.120 7.05 1.23 7.838 6.095 8.658 6.304 7.22 1.23 8.024 6.240 8.807 6.420 7.37 1.25 8.114 6.300 8.955 6.534 7.48 1.27 8.228 6.388 9.050 6.656 7.58 1.27 8.312 6.460 9.159 6.748 7.67 1.28 8.479 6.546 9.299 6.817 7.79 1.32 8.672 6.724 9.494 6.931 7.96 1.35 8.711 6.908 9.698 6.987 8.08 1.36 8.785 7.060 9.857 7.086 8.20 1.37 8.830 7.178 9.942 7.105 8.26 1.37 8.844 7.263 10.006 7.200 8.33 1.35 8.903 7.332 10.105 7.265 8.40 1.36 8.920 7.393 10.230 7.319 8.47 1.39 8.996 7.574 10.390 7.448 8.60 1.38 9.098 7.806 10.490 7.629 8.76 1.33 9.384 7.976 10.763 7.816 8.98 1.38 9.629 8.172 11.070 8.089 9.24 1.41 9.819 8.314 11.308 8.285 9.43 1.44 9.936 8.454 11.426 8.447 9.57 1.42 Page A29 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Metabolic cages group 4: HF + 0.006 % PFHS Time Accumulated food intake (g) Average acc. SD (hr) cage 0103 cage 0107 cage 0203 cage 0207 food intake (g) (g) 6.1 0.211 0.360 0.336 0.424 0.33 0.09 7.4 0.535 0.688 0.784 0.907 0.73 0.16 8.6 0.656 0.881 1.069 1.157 0.94 0.22 9.9 0.844 1.075 1.187 1.292 1.10 0.19 11.1 0.952 1.146 1.318 1.510 1.23 0.24 12.4 1.045 1.282 1.490 1.612 1.36 0.25 13.6 1.181 1.410 1.611 1.682 1.47 0.23 14.9 1.355 1.470 1.719 1.825 1.59 0.22 16.1 1.475 1.518 1.796 1.867 1.66 0.20 17.4 1.612 1.603 1.896 2.072 1.80 0.23 18.6 1.724 1.856 2.060 2.173 1.95 0.20 19.9 1.840 2.142 2.138 2.328 2.11 0.20 21.1 2.066 2.412 2.257 2.463 2.30 0.18 22.4 2.272 2.630 2.440 2.629 2.49 0.17 23.6 2.408 2.775 2.575 2.771 2.63 0.18 24.9 2.588 2.822 2.849 2.957 2.80 0.16 26.1 2.702 2.952 3.090 3.204 2.99 0.22 27.4 2.899 3.350 3.486 3.580 3.33 0.30 28.6 3.247 3.880 3.988 4.074 3.80 0.38 29.9 3.592 4.090 4.394 4.524 4.15 0.41 31.1 3.788 4.209 4.849 4.851 4.42 0.52 32.4 3.849 4.315 5.167 5.095 4.61 0.64 33.6 3.935 4.435 5.255 5.180 4.70 0.63 34.9 4.033 4.530 5.345 5.222 4.78 0.62 36.1 4.112 4.596 5.513 5.308 4.88 0.65 37.4 4.297 4.917 5.702 5.423 5.08 0.62 38.6 4.563 5.184 5.847 5.572 5.29 0.56 39.9 4.733 5.370 5.977 5.836 5.48 0.56 41.1 4.925 5.644 6.195 5.974 5.68 0.55 42.4 5.058 5.862 6.305 6.104 5.83 0.55 43.6 5.163 6.025 6.418 6.208 5.95 0.55 44.9 5.285 6.249 6.561 6.264 6.09 0.56 46.1 5.412 6.429 6.782 6.450 6.27 0.59 47.4 5.592 6.682 7.023 6.601 6.47 0.62 48.6 5.945 6.866 7.303 6.770 6.72 0.57 49.9 6.233 7.064 7.717 6.965 6.99 0.61 51.1 6.686 7.269 8.032 7.365 7.34 0.55 52.4 7.036 7.546 8.318 7.866 7.69 0.54 53.6 7.312 7.920 8.558 8.118 7.98 0.52 54.9 7.437 8.042 8.678 8.400 8.14 0.54 56.1 7.544 8.134 8.803 8.589 8.27 0.56 57.4 7.578 8.166 8.931 8.735 8.35 0.61 58.6 7.667 8.205 9.030 8.832 8.43 0.62 59.9 7.792 8.260 9.197 8.900 8.54 0.63 61.1 7.871 8.338 9.339 9.022 8.64 0.66 62.4 7.998 8.416 9.506 9.156 8.77 0.69 63.6 8.093 8.503 9.620 9.257 8.87 0.70 64.9 8.272 8.585 9.680 9.388 8.98 0.66 66.1 8.345 8.669 9.753 9.484 9.06 0.66 67.4 8.426 8.750 9.904 9.569 9.16 0.69 68.6 8.474 8.848 10.001 9.658 9.25 0.71 69.9 8.596 8.900 10.123 9.772 9.35 0.72 71.1 8.664 9.017 10.340 9.883 9.48 0.77 72.4 8.858 9.148 10.445 10.118 9.64 0.76 73.6 9.226 9.256 10.689 10.283 9.86 0.74 74.9 9.590 9.385 10.994 10.529 10.12 0.76 76.1 9.961 9.515 11.208 10.865 10.39 0.78 77.4 10.204 9.712 11.433 11.104 10.61 0.79 Metabolic cages group 6: HF + 0.003 % PFOS Time Accumulated food intake (g) Average acc. SD (hr) cage 0104 cage 0108 cage 0204 cage 0208 food intake (g) (g) 6.1 0.204 0.119 0.211 0.256 0.20 0.06 7.4 0.535 0.170 0.629 0.547 0.47 0.20 8.6 0.808 0.260 0.916 0.709 0.67 0.29 9.9 0.960 0.518 1.134 0.865 0.87 0.26 11.1 1.185 0.732 1.406 1.047 1.09 0.28 12.4 1.512 0.847 1.618 1.189 1.29 0.35 13.6 1.749 0.889 1.848 1.340 1.46 0.44 14.9 1.940 0.998 2.001 1.406 1.59 0.47 16.1 2.020 1.095 2.100 1.591 1.70 0.46 17.4 2.174 1.216 2.185 1.642 1.80 0.47 18.6 2.406 1.304 2.252 1.767 1.93 0.50 19.9 2.543 1.370 2.270 1.851 2.01 0.51 21.1 2.648 1.558 2.390 1.938 2.13 0.48 22.4 2.810 1.650 2.480 2.082 2.26 0.50 23.6 3.050 1.750 2.546 2.234 2.40 0.55 24.9 3.301 1.898 2.689 2.577 2.62 0.57 26.1 3.423 2.071 2.851 2.815 2.79 0.55 27.4 3.778 2.310 3.211 3.149 3.11 0.60 28.6 4.101 2.641 3.553 3.436 3.43 0.60 29.9 4.314 2.965 3.990 3.637 3.73 0.58 31.1 4.529 3.048 4.316 3.864 3.94 0.66 32.4 4.663 3.078 4.572 3.978 4.07 0.73 33.6 4.720 3.203 4.695 4.070 4.17 0.71 34.9 4.865 3.307 5.002 4.232 4.35 0.77 36.1 4.989 3.385 5.161 4.322 4.46 0.81 37.4 5.264 3.615 5.358 4.501 4.68 0.81 38.6 5.554 3.670 5.466 4.702 4.85 0.87 39.9 5.835 3.735 5.640 4.919 5.03 0.95 41.1 6.067 3.862 5.747 5.126 5.20 0.97 42.4 6.303 3.946 5.878 5.205 5.33 1.03 43.6 6.441 4.055 6.075 5.268 5.46 1.06 44.9 6.584 4.147 6.188 5.421 5.59 1.07 46.1 6.733 4.293 6.320 5.543 5.72 1.07 47.4 7.107 4.400 6.508 5.644 5.91 1.17 48.6 7.672 4.522 6.617 5.846 6.16 1.33 49.9 7.929 4.758 6.854 6.035 6.39 1.34 51.1 8.198 5.100 7.194 6.268 6.69 1.32 52.4 8.427 5.352 7.512 6.465 6.94 1.33 53.6 8.566 5.511 7.721 6.688 7.12 1.32 54.9 8.762 5.603 7.901 6.921 7.30 1.36 56.1 8.892 5.689 8.062 7.030 7.42 1.38 57.4 9.088 5.749 8.139 7.093 7.52 1.43 58.6 9.207 5.852 8.244 7.166 7.62 1.44 59.9 9.378 6.024 8.302 7.200 7.73 1.44 61.1 9.554 6.105 8.386 7.307 7.84 1.48 62.4 9.689 6.228 8.526 7.433 7.97 1.48 63.6 9.839 6.294 8.674 7.524 8.08 1.52 64.9 10.003 6.361 8.830 7.611 8.20 1.57 66.1 10.139 6.478 8.880 7.664 8.29 1.58 67.4 10.256 6.542 8.933 7.717 8.36 1.60 68.6 10.347 6.591 9.006 7.720 8.42 1.62 69.9 10.464 6.718 9.083 7.740 8.50 1.63 71.1 10.593 6.810 9.309 7.807 8.63 1.66 72.4 10.793 6.915 9.451 7.914 8.77 1.71 73.6 11.011 7.103 9.579 8.188 8.97 1.70 74.9 11.264 7.345 9.788 8.977 9.34 1.63 76.1 11.518 7.679 10.107 9.184 9.62 1.61 77.4 11.641 7.878 10.312 9.426 9.81 1.58 Page A30 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 1.000 4 4 1.000 4 4 1.000 4 4 1.000 4 4 0.860 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.886 4 4 1.000 4 4 1.000 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 Accumulated Food intake; p-values Mann Whitney U-test group 1 vs 4 group 1 vs 6 group 3 vs 4 group 3 vs 6 group 4 vs 6 nn p nn p nn p nn p nn p 4 4 0.057 4 4 0.860 4 4 0.057 4 4 0.686 4 4 0.057 4 4 0.200 4 4 0.886 4 4 0.114 4 4 0.886 4 4 0.114 4 4 0.486 4 4 1.000 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 1.000 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.200 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.200 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.343 4 4 0.200 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.343 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.486 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.114 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.057 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.114 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.114 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.114 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.114 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.114 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.057 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.057 4 4 0.686 4 4 0.343 4 4 0.886 4 4 0.686 p<0.05 Page A31 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Feeding bout size measurements Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 cage 0101 0.045 0.043 0.002 0.018 0.017 0.022 0.024 0.023 0.013 0.014 0.023 0.006 0.019 0.008 0.017 0.016 0.038 0.012 0.040 0.029 0.015 0.025 0.011 0.007 0.016 0.031 0.026 0.026 0.018 0.012 0.021 0.008 0.018 0.023 0.009 0.021 0.025 0.010 0.008 0.011 0.013 0.027 0.019 0.007 0.029 0.011 0.002 0.024 0.002 0.008 0.001 0.002 0.017 0.018 0.009 0.019 0.021 0.010 Metabolic cages group 1: HF control Feeding bout size (g) Average feeding cage 0105 cage 0201 cage 0205 bout size (g) 0.036 0.018 0.044 0.036 0.019 0.032 0.025 0.030 0.029 0.034 0.018 0.021 0.002 0.012 0.025 0.014 0.014 0.010 0.008 0.012 0.032 0.053 0.022 0.032 0.013 0.038 0.017 0.023 0.017 0.018 0.013 0.018 0.010 0.005 0.013 0.010 0.005 0.012 0.012 0.011 0.013 0.002 0.003 0.010 0.005 0.078 0.012 0.025 0.014 0.015 0.008 0.014 0.028 0.021 0.014 0.018 0.020 0.000 0.025 0.016 0.005 0.003 0.011 0.009 0.030 0.000 0.016 0.021 0.012 0.005 0.026 0.014 0.011 0.021 0.028 0.025 0.011 0.021 0.009 0.018 0.010 0.000 0.042 0.017 0.002 0.016 0.004 0.012 0.008 0.006 0.003 0.007 0.009 0.008 0.015 0.010 0.016 0.008 0.033 0.018 0.004 0.009 0.010 0.014 0.046 0.011 0.015 0.025 0.025 0.025 0.015 0.023 0.012 0.013 0.010 0.013 0.017 0.010 0.011 0.013 0.014 0.013 0.017 0.016 0.018 0.024 0.017 0.017 0.023 0.021 0.021 0.021 0.020 0.018 0.020 0.020 0.023 0.021 0.017 0.018 0.017 0.012 0.028 0.020 0.014 0.038 0.030 0.027 0.014 0.025 0.029 0.020 0.009 0.008 0.011 0.009 0.009 0.023 0.015 0.015 0.013 0.021 0.016 0.016 0.012 0.015 0.000 0.014 0.012 0.016 0.007 0.014 0.009 0.015 0.018 0.012 0.004 0.013 0.009 0.014 0.020 0.008 0.000 0.010 0.005 0.004 0.008 0.005 0.007 0.008 0.012 0.013 0.002 0.001 0.001 0.002 0.010 0.018 0.003 0.010 0.015 0.009 0.014 0.010 0.017 0.017 0.001 0.009 0.002 0.014 0.021 0.014 0.047 0.019 0.015 0.025 0.012 0.017 0.027 0.016 0.038 0.035 0.020 0.028 0.002 0.013 0.027 0.016 0.010 0.015 0.014 0.012 SD (g) 0.013 0.010 0.014 0.010 0.004 0.015 0.011 0.004 0.004 0.004 0.010 0.035 0.005 0.009 0.011 0.006 0.017 0.009 0.012 0.009 0.018 0.011 0.003 0.004 0.011 0.012 0.016 0.005 0.003 0.003 0.004 0.007 0.002 0.002 0.006 0.007 0.010 0.009 0.001 0.006 0.004 0.011 0.005 0.005 0.011 0.008 0.003 0.008 0.001 0.006 0.006 0.009 0.008 0.015 0.008 0.010 0.011 0.003 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 3: Feeding bout size (g) cage 0102 cage 0106 cage 0202 0.045 0.043 0.032 0.018 0.018 0.038 0.024 0.011 0.033 0.017 0.026 0.025 0.036 0.024 0.008 0.002 0.019 0.026 0.036 0.017 0.018 0.012 0.008 0.024 0.015 0.011 0.019 0.013 0.001 0.013 0.010 0.008 0.015 0.001 0.004 0.028 0.018 0.018 0.024 0.012 0.019 0.009 0.009 0.007 0.025 0.015 0.028 0.036 0.028 0.009 0.021 0.028 0.031 0.046 0.020 0.001 0.027 0.011 0.000 0.024 0.024 0.003 0.016 0.022 0.008 0.012 0.000 0.000 0.023 0.014 0.011 0.010 0.013 0.012 0.015 0.055 0.010 0.031 0.044 0.025 0.028 0.009 0.012 0.032 0.027 0.031 0.012 0.006 0.015 0.011 0.014 0.011 0.020 0.010 0.009 0.010 0.023 0.017 0.018 0.017 0.032 0.017 0.005 0.033 0.024 0.038 0.022 0.025 0.032 0.019 0.032 0.030 0.018 0.020 0.013 0.008 0.014 0.025 0.015 0.011 0.015 0.010 0.023 0.014 0.006 0.004 0.007 0.013 0.015 0.013 0.011 0.007 0.020 0.001 0.022 0.008 0.030 0.022 0.004 0.009 0.007 0.011 0.019 0.020 0.000 0.011 0.005 0.007 0.007 0.006 0.002 0.005 0.013 0.000 0.011 0.009 0.013 0.023 0.018 0.014 0.023 0.010 0.033 0.008 0.041 0.019 0.023 0.022 0.017 0.008 0.021 0.006 0.015 0.010 HF + 0.03 % PFBS Average feeding cage 0206 bout size (g) 0.041 0.040 0.010 0.021 0.014 0.021 0.015 0.021 0.015 0.021 0.020 0.017 0.020 0.023 0.008 0.013 0.011 0.014 0.007 0.009 0.012 0.011 0.005 0.010 0.003 0.016 0.015 0.014 0.020 0.015 0.027 0.027 0.012 0.018 0.029 0.034 0.021 0.017 0.009 0.011 0.029 0.018 0.009 0.013 0.008 0.008 0.018 0.013 0.012 0.013 0.032 0.032 0.016 0.028 0.020 0.018 0.009 0.020 0.003 0.009 0.012 0.014 0.010 0.010 0.016 0.019 0.022 0.022 0.011 0.018 0.024 0.027 0.024 0.027 0.023 0.023 0.000 0.009 0.026 0.019 0.008 0.014 0.017 0.010 0.010 0.011 0.002 0.008 0.012 0.014 0.011 0.018 0.002 0.006 0.010 0.015 0.001 0.004 0.014 0.009 0.006 0.007 0.002 0.006 0.022 0.019 0.014 0.015 0.019 0.025 0.032 0.024 0.017 0.016 0.013 0.011 SD (g) 0.006 0.012 0.010 0.006 0.012 0.010 0.009 0.008 0.004 0.006 0.003 0.012 0.009 0.004 0.009 0.009 0.009 0.008 0.011 0.010 0.011 0.006 0.011 0.004 0.001 0.018 0.012 0.010 0.011 0.005 0.004 0.001 0.003 0.007 0.013 0.007 0.006 0.005 0.006 0.007 0.007 0.006 0.004 0.005 0.010 0.010 0.003 0.005 0.005 0.004 0.005 0.005 0.005 0.006 0.015 0.006 0.006 0.004 Page A32 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 4: HF + 0.006 % PFHS Feeding bout size (g) Average feeding cage 0103 cage 0107 cage 0203 cage 0207 bout size (g) 0.043 0.050 0.060 0.066 0.055 0.019 0.023 0.037 0.037 0.029 0.009 0.023 0.014 0.025 0.018 0.022 0.014 0.011 0.009 0.014 0.007 0.007 0.025 0.021 0.015 0.010 0.024 0.006 0.004 0.011 0.020 0.000 0.015 0.020 0.014 0.010 0.006 0.011 0.001 0.007 0.013 0.005 0.006 0.019 0.011 0.013 0.012 0.007 0.012 0.011 0.011 0.027 0.021 0.005 0.016 0.021 0.040 0.008 0.023 0.023 0.018 0.013 0.014 0.008 0.013 0.016 0.029 0.010 0.021 0.019 0.015 0.009 0.025 0.013 0.016 0.019 0.001 0.024 0.024 0.017 0.009 0.019 0.024 0.030 0.021 0.036 0.063 0.056 0.041 0.049 0.033 0.030 0.044 0.048 0.039 0.024 0.020 0.049 0.049 0.036 0.014 0.012 0.036 0.017 0.020 0.005 0.009 0.019 0.025 0.015 0.010 0.017 0.005 0.000 0.008 0.012 0.000 0.018 0.010 0.010 0.009 0.012 0.013 0.008 0.011 0.028 0.032 0.015 0.007 0.021 0.019 0.040 0.020 0.030 0.027 0.017 0.000 0.012 0.019 0.012 0.022 0.044 0.021 0.011 0.025 0.009 0.015 0.007 0.011 0.011 0.007 0.016 0.010 0.008 0.010 0.012 0.017 0.030 0.019 0.020 0.013 0.035 0.011 0.010 0.017 0.028 0.007 0.029 0.017 0.020 0.037 0.026 0.037 0.017 0.029 0.033 0.015 0.035 0.022 0.026 0.040 0.027 0.027 0.060 0.039 0.030 0.039 0.032 0.026 0.032 0.022 0.025 0.023 0.036 0.027 0.013 0.004 0.001 0.020 0.010 0.008 0.008 0.021 0.016 0.013 0.006 0.002 0.006 0.014 0.007 0.014 0.007 0.018 0.006 0.011 0.004 0.005 0.017 0.007 0.008 0.011 0.008 0.015 0.018 0.013 0.014 0.012 0.014 0.011 0.013 0.017 0.002 0.009 0.009 0.009 0.008 0.015 0.000 0.014 0.009 0.011 0.009 0.022 0.009 0.013 0.006 0.000 0.001 0.008 0.004 0.001 0.010 0.017 0.008 0.009 0.014 0.010 0.013 0.013 0.013 0.006 0.014 0.015 0.013 0.012 0.034 0.013 0.011 0.024 0.021 0.041 0.008 0.035 0.023 0.027 0.034 0.015 0.030 0.025 0.026 0.037 0.016 0.019 0.036 0.027 0.017 0.014 0.025 0.013 0.017 SD (g) 0.010 0.009 0.008 0.006 0.009 0.009 0.009 0.005 0.007 0.003 0.010 0.013 0.004 0.008 0.007 0.011 0.009 0.013 0.009 0.016 0.011 0.009 0.007 0.007 0.002 0.012 0.010 0.009 0.014 0.003 0.004 0.008 0.012 0.010 0.010 0.009 0.016 0.005 0.006 0.009 0.006 0.005 0.006 0.006 0.004 0.002 0.006 0.007 0.006 0.004 0.007 0.002 0.004 0.011 0.015 0.008 0.011 0.005 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 6: HF + 0.003 % PFOS Feeding bout size (g) Average feeding cage 0104 cage 0108 cage 0204 cage 0208 bout size (g) 0.040 0.017 0.037 0.035 0.032 0.026 0.000 0.043 0.025 0.024 0.020 0.010 0.018 0.018 0.017 0.024 0.036 0.025 0.018 0.026 0.022 0.015 0.027 0.013 0.019 0.030 0.009 0.027 0.014 0.020 0.027 0.003 0.014 0.014 0.015 0.008 0.019 0.019 0.008 0.014 0.010 0.001 0.000 0.018 0.007 0.026 0.013 0.009 0.002 0.013 0.014 0.013 0.007 0.012 0.012 0.011 0.008 0.010 0.013 0.011 0.015 0.017 0.011 0.013 0.014 0.019 0.010 0.004 0.014 0.012 0.024 0.013 0.006 0.015 0.015 0.018 0.010 0.023 0.033 0.021 0.013 0.031 0.015 0.033 0.023 0.050 0.013 0.049 0.024 0.034 0.020 0.046 0.029 0.033 0.032 0.027 0.016 0.031 0.011 0.021 0.013 0.006 0.035 0.024 0.020 0.014 0.003 0.023 0.011 0.013 0.010 0.016 0.017 0.012 0.014 0.011 0.012 0.036 0.019 0.020 0.011 0.004 0.004 0.002 0.005 0.039 0.023 0.022 0.023 0.027 0.029 0.007 0.014 0.022 0.018 0.024 0.013 0.012 0.026 0.019 0.023 0.004 0.013 0.014 0.014 0.018 0.016 0.017 0.002 0.013 0.013 0.003 0.011 0.012 0.010 0.020 0.020 0.012 0.013 0.016 0.013 0.010 0.015 0.015 0.013 0.066 0.010 0.020 0.018 0.029 0.026 0.016 0.019 0.012 0.018 0.031 0.029 0.032 0.019 0.028 0.027 0.029 0.031 0.027 0.029 0.021 0.019 0.029 0.023 0.023 0.012 0.014 0.017 0.014 0.014 0.014 0.012 0.016 0.026 0.017 0.019 0.005 0.012 0.005 0.010 0.015 0.006 0.005 0.011 0.009 0.015 0.014 0.014 0.001 0.011 0.014 0.015 0.001 0.006 0.009 0.023 0.010 0.014 0.013 0.015 0.012 0.013 0.014 0.010 0.012 0.016 0.001 0.021 0.008 0.012 0.015 0.017 0.005 0.009 0.012 0.010 0.002 0.005 0.008 0.006 0.012 0.008 0.007 0.001 0.007 0.011 0.003 0.009 0.000 0.006 0.012 0.014 0.006 0.007 0.010 0.009 0.014 0.027 0.003 0.013 0.022 0.008 0.011 0.020 0.015 0.028 0.028 0.021 0.087 0.041 0.027 0.021 0.019 0.015 0.021 0.020 0.031 0.036 0.030 0.029 0.007 0.016 0.013 0.016 0.013 SD (g) 0.010 0.018 0.004 0.008 0.006 0.010 0.010 0.006 0.008 0.010 0.003 0.002 0.003 0.006 0.007 0.010 0.010 0.018 0.011 0.009 0.013 0.008 0.003 0.012 0.004 0.008 0.010 0.007 0.008 0.008 0.005 0.004 0.002 0.025 0.006 0.006 0.002 0.004 0.002 0.006 0.007 0.005 0.007 0.007 0.006 0.002 0.009 0.006 0.004 0.005 0.005 0.004 0.010 0.007 0.031 0.005 0.007 0.004 Page A33 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.886 4 4 0.200 4 4 0.886 4 4 0.343 4 4 0.343 4 4 0.114 4 4 0.886 4 4 0.343 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.200 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.057 4 4 0.686 4 4 0.029 4 4 0.343 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.343 4 4 0.114 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.200 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.460 4 4 0.343 4 4 0.200 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.686 Feeding bout size; p-values Mann Whitney U-test group 1 vs 4 group 1 vs 6 group 3 vs 4 group 3 vs 6 group 4 vs 6 nn p nn p nn p nn p nn p 4 4 0.114 4 4 0.486 4 4 0.057 4 4 0.200 4 4 0.029 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.200 4 4 0.114 4 4 0.486 4 4 0.057 4 4 1.000 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.114 4 4 0.343 4 4 0.686 4 4 0.686 4 4 0.200 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.200 4 4 0.886 4 4 0.029 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.886 4 4 0.486 4 4 0.486 4 4 0.200 4 4 0.486 4 4 1.000 4 4 1.000 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.200 4 4 0.343 4 4 0.200 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.886 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.114 4 4 0.200 4 4 0.486 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.343 4 4 0.686 4 4 0.029 4 4 0.114 4 4 0.114 4 4 1.000 4 4 0.343 4 4 0.114 4 4 0.486 4 4 0.029 4 4 0.114 4 4 0.343 4 4 0.200 4 4 0.486 4 4 0.057 4 4 0.114 4 4 0.343 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.057 4 4 0.886 4 4 0.343 4 4 0.343 4 4 0.686 4 4 0.114 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.200 4 4 0.057 4 4 0.200 4 4 0.029 4 4 0.114 4 4 0.486 4 4 0.200 4 4 0.343 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.343 4 4 0.343 4 4 0.114 4 4 0.343 4 4 0.486 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.686 4 4 0.860 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.486 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.114 4 4 0.029 4 4 0.200 4 4 0.343 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.029 4 4 0.029 4 4 0.886 4 4 0.343 4 4 0.029 4 4 0.343 4 4 0.029 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.114 4 4 1.000 4 4 0.200 4 4 0.886 4 4 0.114 4 4 0.343 4 4 0.140 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.343 4 4 1.000 4 4 0.686 4 4 0.057 4 4 0.686 4 4 0.057 4 4 0.886 4 4 0.057 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.200 4 4 0.029 4 4 0.486 4 4 0.486 4 4 0.200 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.200 4 4 0.886 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.200 4 4 0.343 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.343 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.343 4 4 0.343 4 4 0.486 4 4 0.029 4 4 0.057 4 4 0.114 4 4 0.486 4 4 0.114 4 4 0.114 4 4 0.486 4 4 0.200 4 4 1.000 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.886 4 4 0.686 4 4 1.000 4 4 1.000 4 4 0.057 4 4 0.343 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.114 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.057 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.343 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.114 4 4 0.343 4 4 0.057 4 4 0.886 4 4 0.200 4 4 0.686 4 4 0.114 4 4 0.343 4 4 0.343 p<0.05 Page A34 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Drink intake measurements Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 1: HF control Accumulated drink intake (ml) Average acc. SD cage 0101 cage 0105 cage 0201 cage 0205 drink intake (ml) (ml) 0.205 0.065 0.129 0.230 0.16 0.08 0.720 0.337 0.591 0.580 0.56 0.16 0.946 0.658 0.782 0.846 0.81 0.12 1.053 0.791 0.784 1.083 0.93 0.16 1.209 0.845 0.805 1.297 1.04 0.25 1.284 1.166 0.936 1.334 1.18 0.18 1.375 1.292 1.208 1.481 1.34 0.12 1.602 1.326 1.325 1.610 1.47 0.16 1.716 1.448 1.348 1.624 1.53 0.17 1.799 1.479 1.373 1.702 1.59 0.20 1.836 1.499 1.382 1.806 1.63 0.22 1.898 1.530 1.463 1.826 1.68 0.21 1.981 1.530 1.564 1.863 1.73 0.22 2.002 1.530 1.576 1.891 1.75 0.23 2.049 1.778 1.610 1.976 1.85 0.20 2.213 1.870 1.610 2.024 1.93 0.25 2.280 2.047 1.796 2.024 2.04 0.20 2.465 2.264 1.927 2.098 2.19 0.23 2.714 2.349 2.268 2.291 2.41 0.21 3.045 2.480 2.638 2.436 2.65 0.28 3.250 2.581 2.921 2.691 2.86 0.30 3.375 2.669 3.022 2.949 3.00 0.29 3.523 2.720 3.227 2.967 3.11 0.34 3.640 2.863 3.434 3.036 3.24 0.36 3.734 3.104 3.738 3.252 3.46 0.33 3.796 3.281 3.818 3.353 3.56 0.28 4.053 3.602 4.089 3.434 3.79 0.33 4.358 3.893 4.292 3.680 4.06 0.32 4.508 4.029 4.460 3.765 4.19 0.36 4.524 4.072 4.508 3.832 4.23 0.34 4.654 4.114 4.517 3.885 4.29 0.36 4.716 4.201 4.683 4.004 4.40 0.35 4.784 4.325 4.812 4.170 4.52 0.32 4.966 4.510 4.908 4.342 4.68 0.30 5.096 4.661 5.072 4.499 4.83 0.30 5.140 4.925 5.175 4.538 4.94 0.29 5.200 5.018 5.251 4.664 5.03 0.27 5.343 5.110 5.469 4.848 5.19 0.27 5.465 5.280 5.699 4.922 5.34 0.33 5.530 5.423 5.895 5.037 5.47 0.35 5.694 5.540 6.008 5.175 5.60 0.35 5.892 5.799 6.279 5.348 5.83 0.38 6.146 5.984 6.385 5.428 5.99 0.41 6.297 6.086 6.578 5.481 6.11 0.47 6.399 6.137 6.760 5.557 6.21 0.51 6.604 6.372 6.937 5.612 6.38 0.56 6.604 6.499 7.038 5.653 6.45 0.58 6.695 6.511 7.075 5.773 6.51 0.55 6.796 6.511 7.222 5.865 6.60 0.57 6.812 6.547 7.270 5.865 6.62 0.59 6.812 6.562 7.447 5.900 6.68 0.64 6.812 6.800 7.544 6.056 6.80 0.61 6.942 6.848 7.694 6.148 6.91 0.63 7.192 7.031 7.822 6.297 7.09 0.63 7.311 7.293 7.852 6.387 7.21 0.61 7.535 7.560 8.022 6.608 7.43 0.59 7.639 7.701 8.317 6.845 7.63 0.60 7.875 7.752 8.349 7.043 7.75 0.54 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 3: HF + 0.03 % PFBS Accumulated drink intake (ml) Average acc. SD cage 0102 cage 0106 cage 0202 cage 0206 drink intake (ml) (ml) 0.125 0.077 0.074 0.092 0.09 0.02 0.387 0.392 0.569 0.388 0.43 0.09 0.699 0.496 0.967 0.560 0.68 0.21 0.883 0.510 1.102 0.676 0.79 0.26 0.912 0.691 1.253 0.864 0.93 0.24 0.994 0.800 1.449 1.024 1.07 0.27 1.176 0.824 1.634 1.276 1.23 0.33 1.318 0.834 1.800 1.400 1.34 0.40 1.462 0.848 1.850 1.400 1.39 0.41 1.565 0.861 2.153 1.450 1.51 0.53 1.592 0.872 2.254 1.500 1.55 0.57 1.600 0.880 2.311 1.500 1.57 0.59 1.659 0.880 2.578 1.548 1.67 0.70 1.730 0.923 2.743 1.616 1.75 0.75 1.768 1.003 2.804 1.780 1.84 0.74 1.837 1.013 3.065 1.842 1.94 0.85 1.898 1.024 3.283 1.988 2.05 0.93 2.045 1.078 3.512 2.146 2.20 1.00 2.210 1.179 3.904 2.280 2.39 1.13 2.357 1.194 4.171 2.418 2.53 1.23 2.539 1.205 4.361 2.606 2.68 1.29 2.710 1.216 4.558 2.764 2.81 1.37 2.912 1.232 4.669 2.906 2.93 1.40 2.968 1.245 4.869 2.988 3.02 1.48 3.101 1.302 4.941 3.026 3.09 1.49 3.251 1.360 5.004 3.248 3.22 1.49 3.547 1.386 5.263 3.424 3.41 1.59 3.787 1.453 5.465 3.520 3.56 1.65 3.955 1.557 5.645 3.520 3.67 1.68 4.046 1.664 5.702 3.590 3.75 1.66 4.059 1.725 5.731 3.632 3.79 1.65 4.130 1.776 5.814 3.722 3.86 1.66 4.232 1.864 6.048 3.788 3.98 1.72 4.293 1.925 6.185 3.874 4.07 1.75 4.387 2.098 6.361 4.048 4.22 1.75 4.488 2.166 6.577 4.100 4.33 1.81 4.763 2.238 6.869 4.252 4.53 1.90 5.006 2.384 6.988 4.420 4.70 1.89 5.194 2.560 7.207 4.608 4.89 1.91 5.344 2.587 7.337 4.804 5.02 1.95 5.506 2.672 7.529 5.032 5.18 1.99 5.597 2.782 7.729 5.188 5.32 2.03 5.693 2.838 7.884 5.352 5.44 2.07 5.742 2.869 8.100 5.420 5.53 2.14 5.842 2.960 8.217 5.510 5.63 2.15 6.122 3.056 8.334 5.646 5.79 2.17 6.157 3.131 8.496 5.700 5.87 2.20 6.176 3.146 8.674 5.836 5.96 2.26 6.198 3.157 8.726 5.900 6.00 2.28 6.213 3.168 8.748 5.946 6.02 2.28 6.235 3.224 8.748 6.000 6.05 2.26 6.250 3.272 8.748 6.000 6.07 2.24 6.301 3.341 8.975 6.072 6.17 2.30 6.358 3.434 9.104 6.260 6.29 2.32 6.584 3.462 9.446 6.362 6.46 2.44 6.795 3.486 9.756 6.532 6.64 2.56 6.912 3.550 9.992 6.730 6.80 2.63 6.998 3.640 10.177 6.890 6.93 2.67 Page A35 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Metabolic cages group 4: HF + 0.006 % PFHS Time Accumulated drink intake (ml) Average acc. SD (hr) cage 0103 cage 0107 cage 0203 cage 0207 drink intake (ml) (ml) 6.1 0.131 0.362 0.214 0.306 0.25 0.10 7.4 0.361 0.782 0.880 0.849 0.72 0.24 8.6 0.479 0.996 1.327 1.135 0.98 0.36 9.9 0.574 1.207 1.512 1.254 1.14 0.40 11.1 0.612 1.207 1.718 1.395 1.23 0.46 12.4 0.669 1.248 1.961 1.518 1.35 0.54 13.6 0.768 1.343 1.995 1.558 1.42 0.51 14.9 0.819 1.370 2.085 1.606 1.47 0.53 16.1 0.982 1.377 2.184 1.624 1.54 0.50 17.4 1.113 1.377 2.184 1.694 1.59 0.46 18.6 1.121 1.550 2.297 1.729 1.67 0.49 19.9 1.159 1.758 2.310 1.815 1.76 0.47 21.1 1.290 1.880 2.329 1.971 1.87 0.43 22.4 1.463 1.938 2.520 2.024 1.99 0.43 23.6 1.516 1.955 2.545 2.024 2.01 0.42 24.9 1.592 1.955 2.747 2.081 2.09 0.48 26.1 1.615 1.955 2.982 2.152 2.18 0.58 27.4 1.685 2.072 3.137 2.429 2.33 0.62 28.6 2.048 2.708 3.620 2.974 2.84 0.65 29.9 2.329 2.948 4.053 3.518 3.21 0.74 31.1 2.453 3.074 4.574 3.846 3.49 0.92 32.4 2.512 3.186 4.893 4.052 3.66 1.04 33.6 2.546 3.349 4.998 4.092 3.75 1.05 34.9 2.595 3.451 4.998 4.145 3.80 1.02 36.1 2.645 3.461 5.093 4.312 3.88 1.06 37.4 2.683 3.667 5.187 4.312 3.96 1.06 38.6 2.831 3.856 5.271 4.396 4.09 1.02 39.9 2.945 3.910 5.414 4.576 4.21 1.04 41.1 3.027 4.175 5.527 4.708 4.36 1.05 42.4 3.173 4.352 5.565 4.717 4.45 0.99 43.6 3.192 4.352 5.649 4.752 4.49 1.02 44.9 3.211 4.423 5.712 4.759 4.53 1.03 46.1 3.276 4.498 5.943 4.888 4.65 1.10 47.4 3.416 4.651 6.203 4.950 4.81 1.14 48.6 3.585 4.763 6.422 4.950 4.93 1.16 49.9 3.753 4.859 6.766 5.025 5.10 1.25 51.1 4.051 5.005 7.046 5.350 5.36 1.25 52.4 4.366 5.263 7.388 5.870 5.72 1.27 53.6 4.579 5.632 7.642 6.395 6.06 1.29 54.9 4.579 5.794 7.854 6.743 6.24 1.39 56.1 4.579 5.868 7.892 7.018 6.34 1.44 57.4 4.579 5.930 8.100 7.227 6.46 1.54 58.6 4.699 6.011 8.245 7.454 6.60 1.57 59.9 4.803 6.042 8.392 7.608 6.71 1.60 61.1 4.872 6.106 8.610 7.726 6.83 1.67 62.4 4.940 6.188 8.736 7.806 6.92 1.69 63.6 4.978 6.297 8.736 7.854 6.97 1.67 64.9 5.043 6.351 8.736 7.920 7.01 1.64 66.1 5.088 6.450 8.763 8.004 7.08 1.64 67.4 5.111 6.460 8.904 8.074 7.14 1.69 68.6 5.111 6.504 8.988 8.140 7.19 1.73 69.9 5.172 6.542 9.114 8.175 7.25 1.75 71.1 5.187 6.664 9.284 8.228 7.34 1.79 72.4 5.324 6.725 9.303 8.391 7.44 1.77 73.6 5.596 6.732 9.574 8.532 7.61 1.78 74.9 5.917 6.773 9.811 8.708 7.80 1.78 76.1 6.236 6.943 10.019 9.082 8.07 1.78 77.4 6.335 7.133 10.185 9.297 8.24 1.80 Metabolic cages group 6: HF + 0.003 % PFOS Time Accumulated drink intake (ml) Average acc. SD (hr) cage 0104 cage 0108 cage 0204 cage 0208 drink intake (ml) (ml) 6.1 0.134 0.240 0.151 0.202 0.18 0.05 7.4 0.711 0.288 0.724 0.470 0.55 0.21 8.6 1.078 0.328 1.159 0.616 0.80 0.39 9.9 1.282 0.563 1.269 0.716 0.96 0.37 11.1 1.576 0.813 1.505 0.840 1.18 0.41 12.4 1.877 0.883 1.667 0.976 1.35 0.50 13.6 2.179 0.896 1.818 1.062 1.49 0.61 14.9 2.394 0.941 1.957 1.100 1.60 0.69 16.1 2.567 0.992 2.016 1.216 1.70 0.73 17.4 2.669 1.085 2.016 1.280 1.76 0.73 18.6 2.820 1.146 2.016 1.288 1.82 0.77 19.9 2.948 1.253 2.016 1.360 1.89 0.78 21.1 2.992 1.264 2.016 1.422 1.92 0.78 22.4 2.992 1.278 2.016 1.446 1.93 0.77 23.6 3.012 1.312 2.016 1.560 1.98 0.75 24.9 3.135 1.312 2.088 1.576 2.03 0.81 26.1 3.196 1.338 2.160 1.630 2.08 0.82 27.4 3.344 1.506 2.198 1.794 2.21 0.81 28.6 3.611 1.768 2.362 2.022 2.44 0.82 29.9 3.822 2.093 2.907 2.198 2.75 0.80 31.1 4.039 2.198 3.308 2.390 2.98 0.85 32.4 4.097 2.317 3.438 2.534 3.10 0.83 33.6 4.097 2.394 3.620 2.664 3.19 0.80 34.9 4.097 2.557 3.978 2.808 3.36 0.79 36.1 4.097 2.642 4.104 2.940 3.45 0.77 37.4 4.148 2.720 4.111 3.000 3.49 0.74 38.6 4.423 2.774 4.176 3.134 3.63 0.80 39.9 4.648 2.832 4.363 3.342 3.80 0.85 41.1 4.784 2.864 4.437 3.468 3.89 0.88 42.4 4.944 2.902 4.464 3.480 3.95 0.93 43.6 5.117 3.024 4.529 3.494 4.04 0.95 44.9 5.151 3.024 4.698 3.506 4.09 1.00 46.1 5.156 3.046 4.698 3.550 4.11 0.98 47.4 5.202 3.178 4.698 3.636 4.18 0.93 48.6 5.263 3.205 4.698 3.732 4.22 0.93 49.9 5.535 3.334 4.806 3.858 4.38 0.98 51.1 5.879 3.624 5.123 3.974 4.65 1.04 52.4 5.964 3.898 5.377 4.148 4.85 0.99 53.6 6.137 4.242 5.555 4.378 5.08 0.92 54.9 6.392 4.346 5.767 4.560 5.27 0.98 56.1 6.681 4.536 5.994 4.676 5.47 1.04 57.4 6.871 4.608 6.118 4.778 5.59 1.09 58.6 6.989 4.707 6.311 4.958 5.74 1.09 59.9 7.342 4.891 6.390 5.020 5.91 1.17 61.1 7.431 4.912 6.604 5.212 6.04 1.18 62.4 7.497 5.022 6.772 5.346 6.16 1.17 63.6 7.616 5.152 6.804 5.452 6.26 1.16 64.9 7.616 5.168 6.804 5.516 6.28 1.14 66.1 7.616 5.216 6.804 5.540 6.29 1.12 67.4 7.659 5.222 6.804 5.540 6.31 1.13 68.6 7.701 5.232 6.804 5.540 6.32 1.15 69.9 7.701 5.290 6.804 5.568 6.34 1.12 71.1 7.701 5.347 6.804 5.630 6.37 1.09 72.4 7.701 5.408 6.804 5.676 6.40 1.06 73.6 7.813 5.504 6.944 5.824 6.52 1.06 74.9 8.050 5.678 7.175 5.952 6.71 1.10 76.1 8.109 5.987 7.547 6.098 6.94 1.06 77.4 8.109 6.242 7.686 6.304 7.09 0.95 Page A36 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.343 4 4 0.343 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.860 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.343 Accumulated drink intake; p-values Mann Whitney U-test group 1 vs 4 group 1 vs 6 group 3 vs 4 group 3 vs 6 group 4 vs 6 nn p nn p nn p nn p nn p 4 4 0.343 4 4 0.686 4 4 0.029 4 4 0.029 4 4 0.343 4 4 0.200 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.200 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.686 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.486 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.686 4 4 1.000 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 1.000 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.486 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.486 p<0.05 4 4 0.686 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.686 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.686 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.343 Page A37 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 D4. X activity measurements Metabolic cages group 1: HF control X activity per interval cage 0101 cage 0105 cage 0201 cage 0205 100.7 88.8 134.7 99.5 63.9 227.7 123.3 73.9 67.5 124.2 57.2 69.0 76.1 238.3 182.4 62.8 90.7 71.1 72.9 173.1 88.6 74.3 128.0 80.4 69.1 109.8 54.7 35.5 72.2 28.9 67.7 67.7 14.3 92.9 70.7 46.6 112.4 64.9 119.1 63.5 144.9 81.0 41.1 47.5 74.2 81.1 92.5 166.7 44.1 99.0 49.7 29.3 47.4 71.0 121.8 94.7 88.3 91.9 108.7 57.4 424.8 330.7 375.7 615.8 133.7 276.6 421.4 814.9 331.4 346.2 385.6 228.3 121.9 221.1 257.7 488.8 143.4 139.3 121.1 158.0 133.1 229.4 213.0 217.7 74.2 181.4 129.7 250.7 54.6 182.5 199.9 94.7 172.7 166.9 158.2 119.2 84.7 128.2 53.6 109.3 101.7 122.5 136.4 118.5 53.4 25.2 67.8 54.2 33.9 49.9 56.4 55.9 84.9 47.7 44.7 23.0 28.9 79.6 74.6 119.0 64.8 34.9 59.5 96.2 74.9 64.0 65.7 38.0 69.8 101.4 87.1 72.6 90.4 52.7 46.1 71.2 373.3 279.0 284.9 313.7 337.7 147.0 370.8 371.0 361.6 270.6 276.3 229.2 161.5 365.6 199.2 227.9 164.7 143.5 156.8 202.0 119.6 206.4 153.3 185.1 76.2 117.9 136.6 57.6 82.3 180.9 143.9 106.5 96.9 97.8 154.2 234.6 173.9 144.2 96.5 86.3 90.2 68.3 85.5 90.4 71.4 80.6 87.9 29.3 49.0 185.5 93.7 91.4 94.4 67.4 115.8 76.0 36.2 14.6 23.2 57.4 91.6 118.8 97.4 31.4 42.5 77.5 37.2 106.7 50.0 112.7 53.7 40.3 94.0 43.9 83.9 120.3 186.7 126.5 215.6 235.7 380.3 349.8 374.6 551.9 411.4 223.4 274.0 372.4 246.9 230.0 248.0 185.3 156.4 360.5 270.5 260.9 Average X activity 105.9 122.2 79.5 139.9 102.0 92.8 67.3 59.1 56.1 90.0 78.6 103.6 55.5 83.7 86.6 436.8 411.7 322.9 272.4 140.5 198.3 159.0 132.9 154.3 94.0 119.8 50.2 49.0 50.1 75.5 63.9 60.7 82.7 65.1 312.7 306.6 284.4 238.6 166.8 166.1 97.1 128.4 145.9 125.2 83.6 67.3 104.9 88.4 32.9 84.8 66.0 64.2 85.5 191.1 414.2 320.3 227.6 262.1 SD 19.9 75.0 30.3 84.7 48.3 24.2 31.5 20.3 33.7 29.9 47.5 42.7 30.2 31.9 21.4 125.4 293.4 67.1 155.3 15.2 44.0 75.2 69.6 24.1 32.3 14.3 17.9 10.5 25.7 36.9 25.2 15.8 14.6 19.9 43.1 107.6 55.6 89.0 25.1 37.9 36.5 43.2 64.9 41.1 10.4 26.2 57.5 21.5 18.6 37.5 32.5 32.8 31.7 47.5 92.8 86.7 29.3 83.5 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 3: HF + 0.03 % PFBS X activity per interval Average cage 0102 cage 0106 cage 0202 cage 0206 X activity 92.2 254.5 88.7 185.0 155.1 92.7 238.4 92.6 375.7 199.9 79.1 99.1 110.4 111.4 100.0 131.6 82.0 45.9 221.7 120.3 134.8 124.3 259.8 60.1 144.8 54.7 138.5 174.2 39.8 101.8 134.4 53.3 68.6 182.7 109.8 59.0 82.5 103.6 47.8 73.2 122.4 105.6 146.3 18.4 98.2 84.7 29.6 245.5 93.8 113.4 158.6 172.4 76.4 41.6 112.3 21.0 39.6 50.1 14.5 31.3 93.0 113.8 169.7 118.6 123.8 111.5 160.0 89.9 126.0 121.9 158.6 55.4 169.7 72.4 114.0 454.8 579.9 431.8 455.2 480.4 432.8 621.4 618.1 933.4 651.4 277.4 351.9 422.6 616.8 417.2 325.6 48.8 191.6 130.5 174.1 262.1 373.9 151.6 282.2 267.5 95.4 52.6 238.0 216.4 150.6 127.8 215.5 260.5 148.1 188.0 99.6 135.3 127.1 112.9 118.7 199.2 141.3 217.7 267.4 206.4 135.8 276.8 172.6 73.8 164.8 109.4 98.3 200.9 103.7 128.1 58.6 61.8 132.5 28.6 70.4 61.9 88.5 155.1 26.9 83.1 60.3 164.0 35.5 11.7 67.9 47.7 32.8 87.4 223.3 97.8 69.2 57.8 71.1 68.3 66.6 131.2 97.6 173.2 60.2 115.6 88.0 78.5 143.7 50.2 90.1 120.8 63.9 104.5 30.4 79.9 361.3 203.4 323.6 485.2 343.4 527.6 382.4 442.4 1194.3 636.7 308.7 470.4 298.9 609.6 421.9 201.9 177.0 84.2 327.0 197.5 332.9 80.5 218.2 274.2 226.5 117.4 102.8 219.6 242.1 170.5 136.8 300.6 165.3 167.6 192.6 121.2 113.3 113.2 309.0 164.2 177.5 352.3 305.4 374.6 302.5 236.1 139.3 186.5 212.9 193.7 70.6 171.6 92.8 28.1 90.8 102.9 131.2 71.9 161.7 116.9 72.1 72.4 43.7 9.3 49.4 62.0 63.4 125.6 266.9 129.5 130.8 49.7 53.5 23.2 64.3 75.2 27.2 84.9 221.5 102.2 84.0 116.4 51.9 45.1 74.4 63.1 82.3 73.4 35.7 63.6 129.7 62.8 151.3 92.7 109.1 201.6 143.3 125.5 219.4 172.5 429.8 396.0 387.1 405.0 404.5 278.5 493.7 280.0 701.6 438.5 270.4 175.4 217.9 407.2 267.7 256.5 298.8 295.6 603.1 363.5 SD 79.9 135.9 15.0 76.2 83.5 64.9 60.0 24.9 55.7 92.5 63.4 16.4 32.6 29.4 58.5 67.2 207.6 145.7 116.7 91.3 90.6 61.2 15.8 52.1 85.1 48.8 44.0 54.2 67.1 86.8 6.0 48.1 39.2 40.8 116.1 376.5 147.8 100.1 108.0 70.6 73.4 96.6 88.2 41.5 60.2 38.4 29.9 96.3 46.3 83.4 32.8 20.2 39.2 45.1 18.4 202.5 100.8 160.9 Page A38 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 4: HF + 0.006 % PFHS X activity per interval Average cage 0103 cage 0107 cage 0203 cage 0207 X activity 75.4 118.1 92.7 258.6 136.2 91.1 61.6 100.7 151.8 101.3 32.3 128.9 85.4 124.6 92.8 88.7 58.6 66.1 53.7 66.8 56.0 47.9 89.9 144.3 84.5 79.3 139.0 66.9 185.7 117.7 103.8 31.0 43.1 57.7 58.9 51.9 53.5 153.0 95.1 88.4 65.2 199.5 53.3 90.9 102.2 83.7 172.4 74.7 34.6 91.4 28.4 46.4 42.0 124.4 60.3 59.3 94.9 103.2 76.9 83.6 108.0 72.6 51.6 57.3 72.4 59.8 39.3 122.1 273.1 123.6 71.2 76.4 74.7 95.4 79.4 484.3 589.1 168.2 410.3 413.0 821.1 605.3 368.9 719.7 628.8 418.6 371.6 219.7 566.0 394.0 282.1 99.3 260.1 379.5 255.3 175.2 215.2 227.2 177.6 198.8 203.6 190.6 144.7 107.9 161.7 92.2 84.7 83.6 169.4 107.5 146.6 178.0 157.2 185.6 166.9 85.3 14.1 101.8 125.7 81.7 173.4 94.0 82.6 336.9 171.7 141.5 111.9 103.3 374.0 182.7 97.6 65.8 82.8 63.6 77.5 46.4 28.1 47.1 140.5 65.5 54.7 69.4 73.6 65.1 65.7 47.0 47.6 44.3 73.3 53.1 79.7 55.9 49.8 76.8 65.6 33.1 77.6 137.3 47.2 73.8 141.3 169.8 67.1 187.9 141.5 148.8 21.7 77.9 55.3 75.9 211.0 139.5 118.3 271.5 185.1 555.2 277.4 176.1 489.8 374.6 227.0 219.5 308.1 548.7 325.8 192.2 120.1 214.5 482.5 252.3 145.5 229.0 129.1 229.2 183.2 159.5 85.7 211.4 146.7 150.8 114.2 96.2 119.5 189.7 129.9 64.4 84.0 123.6 69.4 85.4 116.3 161.9 141.4 211.7 157.8 59.8 24.8 79.1 127.3 72.8 98.7 39.5 90.8 97.0 81.5 97.6 87.9 60.4 116.3 90.6 97.9 39.2 108.8 93.5 84.9 87.3 120.3 55.8 55.3 79.7 69.4 44.1 120.1 211.5 111.3 63.4 28.1 69.2 83.9 61.2 34.3 79.2 92.1 126.5 83.0 77.4 110.7 103.2 151.6 110.7 54.9 103.1 77.5 66.3 75.5 121.3 183.4 84.3 76.1 116.3 262.2 613.3 156.4 233.3 316.3 393.8 463.2 326.7 686.7 467.6 152.2 146.0 423.4 234.1 238.9 160.4 290.0 246.2 300.4 249.3 SD 83.5 37.6 44.8 15.5 43.8 55.2 31.9 47.5 66.7 58.1 43.4 19.6 25.4 105.7 10.9 178.9 194.4 142.7 116.2 26.3 43.9 41.5 18.1 48.0 117.3 128.6 15.9 50.8 8.1 13.6 14.9 46.2 53.2 53.8 69.9 177.7 153.9 158.7 53.4 51.6 41.1 26.8 40.5 42.8 28.2 23.3 31.1 30.9 73.9 23.7 38.1 30.8 20.6 48.9 203.0 156.3 129.4 63.7 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 6: HF + 0.003 % PFOS X activity per interval Average cage 0104 cage 0108 cage 0204 cage 0208 X activity 152.1 223.2 114.3 130.4 155.0 136.3 217.5 107.7 211.5 168.3 37.5 132.4 78.6 35.1 70.9 120.1 164.6 61.9 77.3 106.0 165.9 124.0 96.1 64.6 112.7 64.9 56.9 76.3 42.0 60.0 99.3 51.5 54.9 136.8 85.6 66.3 168.5 118.2 51.8 101.2 35.2 74.0 67.3 72.4 62.2 29.9 94.3 55.5 23.5 50.8 75.5 152.4 123.9 48.4 100.1 94.7 30.6 41.5 67.2 58.5 61.8 90.8 77.4 96.6 81.7 41.2 176.7 52.9 68.4 84.8 154.0 98.7 119.9 115.8 122.1 248.5 353.9 134.8 128.7 216.5 432.4 353.1 703.1 283.1 442.9 225.6 313.2 437.8 367.4 336.0 250.2 403.1 203.5 248.2 276.3 130.8 117.3 152.3 164.3 141.2 194.5 243.2 125.1 138.7 175.4 136.1 54.8 78.9 156.4 106.6 138.1 137.0 183.6 133.4 148.0 250.1 184.4 140.2 133.4 177.0 164.8 39.0 140.4 63.1 101.8 111.8 88.9 133.9 108.8 110.9 81.0 99.0 41.5 98.2 79.9 64.7 12.4 52.8 123.7 63.4 29.8 48.0 55.2 50.8 46.0 36.2 141.9 24.8 28.7 57.9 73.6 35.9 147.9 112.9 92.6 75.9 45.5 63.5 32.1 54.3 42.3 63.0 57.3 78.0 60.2 38.4 84.7 46.8 106.0 69.0 140.0 152.0 217.4 78.4 147.0 405.1 292.8 232.2 226.6 289.2 131.5 332.3 228.9 252.9 236.4 212.6 410.7 208.7 235.1 266.8 114.7 310.1 107.5 227.4 189.9 136.0 161.8 93.3 83.1 118.6 198.1 73.7 158.3 108.8 134.7 89.1 134.6 91.5 166.0 120.3 136.5 126.4 110.9 175.2 137.3 180.5 105.8 129.4 141.3 139.3 85.9 71.5 90.4 116.7 91.1 56.8 151.5 70.7 107.5 96.6 74.8 40.4 70.0 67.5 63.2 42.2 130.3 32.1 72.8 69.4 26.7 24.3 85.4 91.4 57.0 74.5 80.0 53.0 61.2 67.2 59.1 27.3 68.7 34.4 47.4 57.3 101.8 40.5 102.7 75.6 55.0 80.3 57.9 55.6 62.2 106.5 43.9 112.9 98.0 90.3 193.8 308.6 110.3 109.8 180.6 256.2 142.5 208.6 290.9 224.6 131.1 305.9 223.4 367.3 256.9 203.7 407.8 178.4 337.2 281.8 SD 48.0 54.7 45.6 46.2 43.0 14.4 40.5 53.2 18.2 32.1 46.8 28.6 15.5 62.3 23.2 106.9 183.9 89.5 87.3 21.1 54.3 47.6 23.8 53.7 60.3 18.4 26.9 46.0 11.2 56.2 48.5 19.3 14.8 31.9 57.0 82.9 82.7 96.7 97.1 36.8 54.7 36.9 27.4 31.2 18.9 42.4 15.5 44.2 36.4 12.3 19.7 31.6 12.1 31.5 94.0 64.3 102.5 109.1 Page A39 of A172 Appendix to TNO-report of 3M-02 Final version Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.886 4 4 0.343 4 4 0.343 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.486 4 4 0.686 4 4 0.200 4 4 1.000 4 4 0.486 4 4 0.029 4 4 0.057 4 4 0.200 4 4 0.886 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.486 4 4 0.057 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.200 4 4 0.200 4 4 0.686 4 4 0.486 4 4 0.343 4 4 1.000 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.886 4 4 0.684 4 4 0.686 4 4 0.029 4 4 0.114 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.029 4 4 0.686 4 4 0.057 4 4 0.114 4 4 0.886 4 4 0.114 4 4 0.114 4 4 1.000 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.886 4 4 0.486 group 1 vs 4 nn p 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.200 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.029 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.057 4 4 0.486 4 4 0.686 4 4 0.114 4 4 0.686 4 4 0.343 4 4 0.343 4 4 1.000 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.029 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.114 4 4 0.343 4 4 0.200 4 4 1.000 4 4 0.686 4 4 0.057 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.057 4 4 0.343 4 4 0.200 4 4 0.686 4 4 1.000 X activity; p-values Mann Whitney U-test group 1 vs 6 group 3 vs 4 group 3 vs 6 nn p nn p nn p 4 4 0.114 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.343 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.057 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.114 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.343 4 4 0.686 4 4 0.200 4 4 0.029 4 4 0.200 4 4 0.343 4 4 0.057 4 4 0.057 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.057 4 4 0.886 4 4 1.000 4 4 0.057 4 4 0.886 4 4 0.029 4 4 0.686 4 4 0.886 4 4 0.200 4 4 1.000 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.486 4 4 0.200 4 4 1.000 4 4 0.343 4 4 0.114 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.486 4 4 0.114 4 4 0.200 4 4 0.886 4 4 0.029 4 4 0.057 4 4 0.686 4 4 0.029 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.486 4 4 1.000 4 4 0.200 4 4 0.343 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.486 4 4 0.486 4 4 0.343 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.686 4 4 0.343 4 4 0.114 4 4 0.114 4 4 0.343 4 4 0.200 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.029 4 4 0.114 4 4 0.057 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.343 4 4 0.343 4 4 0.114 4 4 0.486 4 4 0.686 4 4 0.200 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.200 4 4 0.686 4 4 0.343 4 4 0.486 4 4 0.200 4 4 0.343 4 4 0.886 4 4 0.200 4 4 0.686 4 4 0.886 4 4 0.057 4 4 0.029 4 4 0.686 4 4 0.029 4 4 0.114 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.343 4 4 0.486 4 4 0.200 4 4 0.200 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.486 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.057 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.057 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.029 4 4 0.343 4 4 0.029 4 4 0.200 4 4 0.886 4 4 0.114 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.343 4 4 0.686 group 4 vs 6 nn p 4 4 0.486 4 4 0.114 4 4 0.886 4 4 0.200 4 4 0.343 4 4 0.057 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.029 4 4 0.114 4 4 0.200 4 4 0.686 4 4 0.886 4 4 0.029 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.029 4 4 0.343 4 4 0.343 4 4 0.686 4 4 0.886 4 4 0.057 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.057 4 4 1.000 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.114 4 4 0.486 4 4 0.057 4 4 0.886 4 4 1.000 4 4 0.343 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.200 4 4 0.114 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.029 4 4 1.000 4 4 0.686 04-12-2006 p<0.05 Page A40 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 D5. O2 consumption,CO2 production and respiratory exchange ratio measurements O2 consumption Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 1: HF control VO2 (ml/h/kg mouse) Average VO2 SD cage 0101 cage 0105 cage 0201 cage 0205 (ml/h/kg mouse) 3055 3609 3572 3183 3355 278 2909 3769 3278 2987 3236 389 2786 3499 3070 3028 3096 296 3051 3540 3607 2913 3278 347 2741 3200 2943 3204 3022 224 2814 2966 3194 2903 2969 162 2636 3102 2744 2661 2786 216 2685 2756 2766 2705 2728 39 2399 2814 2770 2676 2665 186 2811 2699 2874 2756 2785 75 3115 2850 2610 2730 2826 216 2766 2917 2939 3098 2930 136 2602 2990 2628 2542 2691 203 2743 2708 2828 2892 2793 83 3001 2983 2979 2785 2937 102 3803 3693 3803 3939 3809 101 2959 3356 3539 3857 3428 375 3391 3518 3497 3027 3358 228 2921 3300 3385 3544 3287 264 2968 3266 3115 2992 3085 136 2981 3263 3316 3042 3150 164 2763 3180 3307 3199 3112 239 2742 3182 3401 2921 3062 290 3160 3090 3203 2842 3074 162 2844 3097 2702 2680 2831 192 2831 2938 3274 2716 2940 240 2671 2564 2826 2528 2647 134 2400 2696 2776 2658 2633 163 2690 2672 2630 2450 2611 110 2344 2796 2790 2828 2689 231 2542 2523 2642 2639 2587 63 2549 2679 2872 2558 2665 151 2757 2884 3000 2753 2849 118 2937 2860 2704 2758 2815 104 3585 3535 3750 3397 3566 146 3458 3277 3756 3348 3460 211 3320 3475 3528 3083 3352 199 2985 3469 3327 3112 3223 216 2933 3202 3213 3143 3123 130 3066 3289 3231 3090 3169 108 2657 3247 3191 2815 2977 287 2818 3230 3258 2922 3057 220 2822 2877 3274 3079 3013 206 2899 3156 3119 2652 2957 233 2683 2820 2794 2648 2736 84 2389 2882 2765 2368 2601 262 2451 2707 2939 2554 2663 212 2727 2568 2982 2495 2693 216 2399 2276 2392 2563 2407 118 2588 2697 2816 2448 2637 157 2392 2652 2564 2667 2569 126 2532 2656 2612 2506 2576 70 2612 2511 2816 2723 2666 132 3115 3156 3363 3306 3235 119 3315 3487 3910 3443 3539 258 3280 3320 3488 3288 3344 98 2917 3251 3373 2915 3114 234 2855 3337 3389 3106 3172 244 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 3: HF + 0.03 % PFBS VO2 (ml/h/kg mouse) Average VO2 SD cage 0102 cage 0106 cage 0202 cage 0206 (ml/h/kg mouse) 3472 3369 3160 3514 3379 158 3395 3409 3368 3614 3446 113 3050 3203 3449 3293 3249 167 3172 2947 3117 3489 3181 226 3251 3125 3816 3179 3343 320 2926 3060 3395 2899 3070 228 3037 2695 3096 3070 2975 188 2788 2851 3097 2552 2822 224 2871 2875 3325 2576 2912 309 2799 2652 3518 2756 2931 396 3056 3007 3074 2662 2950 194 2601 2709 3117 2514 2735 267 2805 2795 3413 2879 2973 296 2750 2921 3103 2867 2910 147 3171 2587 3536 2788 3020 420 3833 3748 4154 3592 3832 237 3636 3978 4043 3820 3869 181 3312 3246 3549 3655 3441 193 3416 2692 3359 3071 3135 332 3230 3470 3388 3377 3366 100 2869 2582 3516 3131 3025 397 2856 2876 3552 3029 3078 325 2994 2690 3323 2895 2975 264 3079 2731 3477 3112 3100 305 2968 3042 3324 2751 3022 237 3173 2525 3366 2900 2991 364 2704 2513 3166 2538 2730 302 2718 2533 3179 2470 2725 320 2628 2616 3017 2540 2700 215 2607 2201 3282 3134 2806 496 2730 2354 3253 2736 2768 369 2797 2485 3350 2734 2842 365 2698 2627 3198 2680 2801 267 3046 2525 3342 2752 2916 355 3577 3124 3851 3637 3547 306 3845 3499 3852 3908 3776 187 3451 3441 3668 3515 3519 105 3181 2990 3312 3294 3194 148 3346 2800 3445 3165 3189 284 3086 2879 3554 3289 3202 289 3090 3022 3415 3186 3178 172 2863 2881 3403 3262 3102 272 3087 3127 3591 3313 3279 230 3056 2850 3467 3039 3103 260 2671 3145 3078 2635 2882 266 2444 2789 2892 2715 2710 191 2399 2700 2935 2380 2603 265 2342 2391 3212 2899 2711 418 2545 2295 2815 2392 2512 227 2404 2324 3006 2846 2645 332 2463 2660 2803 2507 2608 155 2455 2428 2845 2482 2553 196 2691 2326 3266 2798 2770 387 3248 2864 3489 3160 3190 258 3502 3251 4078 3573 3601 346 3217 3329 3610 3467 3406 170 3102 2931 3421 3432 3222 247 3005 2951 3571 3348 3219 293 Page A41 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 4: HF + 0.006 % PFHS VO2 (ml/h/kg mouse) Average VO2 SD cage 0103 cage 0107 cage 0203 cage 0207 (ml/h/kg mouse) 3575 3707 3794 3663 3685 91 3438 3450 3743 3572 3551 141 3276 3564 3720 3214 3443 240 3298 3287 3705 3019 3327 283 3219 3026 3779 3239 3316 323 3263 3409 3672 3345 3422 177 3253 2782 3223 2912 3043 233 3148 2910 3420 2872 3088 253 3131 3357 3169 2875 3133 198 3100 3284 3191 2771 3086 224 3016 2897 2977 2999 2972 53 3076 3165 3223 2870 3084 155 3085 2998 2948 2834 2966 105 3053 2966 3152 3341 3128 161 3230 3150 3288 3059 3182 99 4037 4423 3777 3640 3969 345 4262 4267 4205 4105 4210 75 3794 3878 3828 3747 3812 55 3514 3369 3957 3750 3648 260 3148 3529 3640 3236 3388 234 3395 3172 3650 3219 3359 216 3149 2853 3984 3250 3309 480 3169 3211 3921 3297 3399 352 2970 2429 3604 3069 3018 481 3160 2889 3350 3309 3177 209 3016 3030 3543 3283 3218 249 2941 2788 3214 2655 2899 240 2850 2683 3085 2885 2876 165 2941 2931 3210 2530 2903 280 2907 2730 2981 2725 2836 128 2942 2681 2965 2754 2835 140 2796 2997 3440 2692 2981 331 3066 3271 3046 3180 3141 105 3205 2865 3259 2689 3005 273 3687 3511 3496 3092 3446 252 4214 3964 4094 3677 3987 230 3775 3687 4344 3700 3876 314 3674 3386 3911 3750 3680 220 3475 3563 3848 3319 3551 222 3342 2927 3707 3329 3326 319 3292 3075 3560 3360 3322 200 3037 2999 3737 3049 3206 355 3252 3141 3706 3142 3310 269 2949 2581 3450 2942 2980 357 2998 2658 3240 2765 2915 259 3028 2847 3009 2797 2920 116 2891 2517 3312 2674 2848 345 2898 2895 3049 2635 2869 172 2599 2387 3169 3032 2797 366 2763 2486 2988 2671 2727 209 2667 2703 3097 2657 2781 211 2847 2910 3004 2767 2882 100 2862 2964 3025 2614 2866 181 3237 3292 3296 2919 3186 180 3789 4002 3796 3425 3753 240 3950 3705 4281 3810 3937 251 3367 2971 4117 3470 3481 475 3409 3211 3938 3360 3480 317 Metabolic cages group 6: HF + 0.003 % PFOS Time VO2 (ml/h/kg mouse) Average VO2 SD (hr) cage 0104 cage 0108 cage 0204 cage 0208 (ml/h/kg mouse) 6.1 3936 4321 3970 3628 3963 283 7.4 3779 3980 3795 3648 3800 136 8.6 3334 3807 3674 3379 3549 229 9.9 3695 3783 3579 3441 3624 148 11.1 3866 3646 3646 3141 3575 307 12.4 3633 3225 3349 3124 3333 220 13.6 3766 3107 3222 3167 3315 304 14.9 3592 3471 3249 3007 3330 258 16.1 3335 2903 3089 3039 3091 180 17.4 3212 3171 2998 2777 3040 198 18.6 3726 3504 3191 2894 3329 364 19.9 3676 2963 2911 3007 3139 360 21.1 3305 3099 2949 3046 3100 150 22.4 3297 3319 2980 3014 3152 180 23.6 3731 3204 3078 3199 3303 291 24.9 4099 4024 3349 3392 3716 400 26.1 4351 3972 4050 3767 4035 242 27.4 3752 3870 4131 3917 3918 158 28.6 3941 4064 3600 3773 3845 202 29.9 3747 3497 3405 3448 3524 153 31.1 3948 3675 3488 3364 3619 254 32.4 3818 3178 3407 3492 3474 265 33.6 3918 3425 3579 3323 3561 260 34.9 3886 3428 3396 3234 3486 280 36.1 3850 3073 3304 3074 3325 366 37.4 3591 3040 3332 3159 3280 239 38.6 3316 3011 2944 3124 3099 163 39.9 3267 2725 2955 3142 3022 236 41.1 3116 2830 2972 3019 2984 119 42.4 3148 3212 2800 2929 3022 191 43.6 3309 2822 3150 3182 3116 208 44.9 3392 2842 2934 2916 3021 250 46.1 3310 2932 3103 3085 3108 155 47.4 3301 2919 2962 3075 3064 171 48.6 3857 3307 3541 3244 3487 278 49.9 4409 3774 3676 3620 3870 365 51.1 3834 3995 3818 3874 3880 80 52.4 4037 3881 3610 3592 3780 217 53.6 3747 3731 3514 3608 3650 110 54.9 3907 3451 3525 3338 3555 247 56.1 3965 3167 3620 3404 3539 339 57.4 3733 3412 3604 3462 3553 145 58.6 3792 3344 3376 3424 3484 208 59.9 3870 3327 3465 3392 3513 245 61.1 3598 3153 3213 3205 3292 205 62.4 3697 3167 3007 3175 3262 301 63.6 3151 2771 3077 2885 2971 174 64.9 3203 2934 2705 3044 2972 209 66.1 2953 2527 2901 2845 2806 192 67.4 3241 2925 2873 2718 2939 220 68.6 3204 2665 2843 2776 2872 233 69.9 3161 2950 2659 3014 2946 211 71.1 3089 2860 2748 2780 2869 154 72.4 3570 2917 2989 3115 3148 293 73.6 3761 3686 3194 3269 3477 287 74.9 3836 3329 3493 3677 3584 220 76.1 3648 3583 3465 3833 3632 154 77.4 3736 3613 3363 3688 3600 166 Page A42 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.200 4 4 0.886 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.200 4 4 0.886 4 4 1.000 4 4 0.114 4 4 0.686 4 4 0.686 4 4 0.057 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.343 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.057 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.114 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.343 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.334 4 4 0.886 group 1 vs 4 nn p 4 4 0.057 4 4 0.343 4 4 0.114 4 4 0.886 4 4 0.114 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.029 4 4 0.114 4 4 0.343 4 4 0.343 4 4 0.114 4 4 0.029 4 4 0.029 4 4 0.886 4 4 0.029 4 4 0.029 4 4 0.200 4 4 0.114 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.057 4 4 0.114 4 4 0.200 4 4 0.114 4 4 0.200 4 4 0.686 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.486 4 4 0.486 4 4 0.057 4 4 0.029 4 4 0.057 4 4 0.029 4 4 0.343 4 4 0.114 4 4 0.686 4 4 0.200 4 4 1.000 4 4 0.486 4 4 0.114 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.686 4 4 0.057 4 4 0.029 4 4 0.114 4 4 0.686 4 4 0.486 4 4 0.029 4 4 0.200 4 4 0.200 VO2, p-values Mann Whitney U-test group 1 vs 6 group 3 vs 4 group 3 vs 6 nn p nn p nn p 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.057 4 4 0.200 4 4 0.029 4 4 0.114 4 4 0.343 4 4 0.114 4 4 0.200 4 4 0.486 4 4 0.057 4 4 0.114 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.114 4 4 0.200 4 4 0.029 4 4 0.686 4 4 0.029 4 4 0.029 4 4 0.114 4 4 0.057 4 4 0.029 4 4 0.200 4 4 0.200 4 4 0.114 4 4 0.486 4 4 0.486 4 4 0.057 4 4 0.486 4 4 0.200 4 4 0.486 4 4 0.114 4 4 0.200 4 4 0.057 4 4 0.486 4 4 0.343 4 4 0.029 4 4 0.114 4 4 0.114 4 4 0.029 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.057 4 4 0.029 4 4 0.486 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.057 4 4 0.029 4 4 0.029 4 4 0.886 4 4 0.114 4 4 0.029 4 4 0.200 4 4 0.114 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.057 4 4 0.200 4 4 0.029 4 4 0.029 4 4 0.686 4 4 0.200 4 4 0.114 4 4 0.486 4 4 0.200 4 4 0.114 4 4 0.486 4 4 0.486 4 4 0.029 4 4 0.343 4 4 0.200 4 4 0.057 4 4 0.486 4 4 0.200 4 4 0.029 4 4 0.686 4 4 0.114 4 4 0.057 4 4 1.000 4 4 0.686 4 4 0.029 4 4 0.686 4 4 0.200 4 4 0.057 4 4 0.686 4 4 0.200 4 4 0.057 4 4 0.200 4 4 0.200 4 4 0.057 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.114 4 4 0.200 4 4 0.886 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.029 4 4 0.486 4 4 0.200 4 4 0.114 4 4 0.486 4 4 0.200 4 4 0.029 4 4 0.686 4 4 0.029 4 4 0.029 4 4 0.686 4 4 0.200 4 4 0.029 4 4 0.486 4 4 0.200 4 4 0.029 4 4 0.886 4 4 0.029 4 4 0.029 4 4 0.140 4 4 0.029 4 4 0.114 4 4 0.486 4 4 0.114 4 4 0.200 4 4 0.886 4 4 0.486 4 4 0.057 4 4 0.343 4 4 0.114 4 4 0.057 4 4 0.886 4 4 0.343 4 4 0.057 4 4 0.343 4 4 0.114 4 4 0.029 4 4 0.057 4 4 0.057 4 4 0.114 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.057 4 4 0.029 4 4 0.200 4 4 0.029 4 4 0.486 4 4 0.029 4 4 0.057 4 4 0.343 4 4 0.057 group 4 vs 6 nn p 4 4 0.200 4 4 0.057 4 4 0.486 4 4 0.200 4 4 0.486 4 4 0.486 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.200 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.686 4 4 0.200 4 4 0.486 4 4 0.343 4 4 0.200 4 4 1.000 4 4 0.486 4 4 0.200 4 4 0.343 4 4 0.686 4 4 0.200 4 4 0.114 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.886 4 4 0.486 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.200 4 4 0.114 4 4 0.200 4 4 0.114 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.114 4 4 0.114 4 4 0.486 4 4 0.486 p<0.05 Page A43 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 CO2 production Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 1: HF control VCO2 (ml/h/kg mouse) Average VCO2 SD cage 0101 cage 0105 cage 0201 cage 0205 (ml/h/kg mouse) 2929 3110 3620 2845 3126 348 2978 3408 3399 2787 3143 311 2781 3142 3042 2814 2945 176 2963 3025 3464 2655 3027 333 2773 2718 2664 2841 2749 76 2721 2759 3107 2565 2788 229 2478 2619 2736 2379 2553 157 2629 2308 2708 2391 2509 190 2418 2501 2567 2267 2438 130 2713 2207 2650 2315 2471 248 3008 2361 2357 2233 2490 351 2486 2419 2757 2474 2534 152 2362 2465 2357 2041 2306 184 2412 2231 2731 2440 2453 207 2563 2617 2602 2402 2546 99 3454 3237 3386 3369 3361 91 2775 2971 3388 3044 3044 256 3139 3105 3283 2578 3026 309 2804 2718 3414 3120 3014 318 2860 2620 3100 2592 2793 238 2861 2568 3243 2750 2856 285 2563 2568 3137 2730 2750 270 2599 2436 2976 2197 2552 327 2801 2505 2827 2280 2603 260 2500 2421 2436 2334 2423 68 2517 2267 2933 2241 2490 321 2627 2362 2813 2191 2498 276 2470 2518 2873 2345 2552 227 2748 2440 2656 2095 2485 290 2281 2470 2655 2354 2440 163 2448 2244 2505 2168 2341 161 2369 2400 2829 2209 2451 265 2594 2551 2874 2334 2588 222 2662 2631 2543 2510 2587 71 3258 3226 3635 3092 3303 233 3054 2903 3311 2877 3036 199 3010 3083 3407 2777 3069 261 2744 3012 3274 2839 2967 233 2610 2778 2975 2727 2773 152 2547 2717 3158 2521 2736 295 2403 2640 2997 2341 2595 297 2581 2784 3114 2383 2715 312 2571 2393 2915 2423 2576 239 2631 2574 2798 2088 2523 305 2574 2152 2597 2128 2363 258 2295 2478 2429 1822 2256 299 2211 2122 2445 1985 2191 193 2536 2019 2461 1914 2232 312 2132 1693 1936 2008 1943 185 2274 2161 2434 1765 2159 285 1943 2025 2320 2073 2090 162 1969 2239 2420 1970 2150 221 2228 1950 2546 2267 2248 244 2767 2843 3065 2691 2841 162 2844 3169 3474 2866 3088 297 2927 3054 3241 2818 3010 182 2559 2840 3150 2531 2770 289 2530 2822 2961 2683 2749 185 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 3: HF + 0.03 % PFBS VCO2 (ml/h/kg mouse) Average VCO2 SD cage 0102 cage 0106 cage 0202 cage 0206 (ml/h/kg mouse) 3252 3003 2825 3076 3039 177 3223 3131 3321 3262 3234 80 2920 2824 3352 2920 3004 236 2958 2601 2985 2986 2883 188 3064 2849 3475 2676 3016 345 2699 2726 3161 2520 2776 272 2801 2151 2872 2743 2642 331 2572 2481 2697 2060 2452 276 2714 2400 3020 2190 2581 363 2585 2148 3083 2324 2535 407 2574 2524 2658 2156 2478 222 2164 2181 2852 2015 2303 374 2411 2426 3150 2209 2549 413 2300 2522 2630 2256 2427 179 2736 2188 3094 2372 2598 402 3201 3308 3794 3129 3358 300 3254 3232 3611 3130 3307 210 2985 2775 3335 3196 3073 245 3014 2175 3300 2743 2808 480 2775 2675 3305 2895 2913 277 2439 1920 3161 2671 2548 515 2504 2269 3092 2606 2618 346 2610 2010 2889 2378 2472 372 2617 2140 2941 2590 2572 329 2477 2465 2788 2208 2485 237 2899 1937 2859 2524 2555 445 2685 2169 3039 2355 2562 383 2683 2199 3043 2262 2547 394 2621 2408 2877 2293 2550 257 2474 1986 2836 2555 2463 354 2507 2197 2699 2100 2376 276 2548 2105 2784 2263 2425 301 2467 2244 2683 2211 2401 219 2764 2233 3009 2456 2615 341 3042 2866 3422 3123 3113 232 3429 3157 3477 3172 3309 168 3211 3074 3245 2974 3126 125 3047 2715 2949 2936 2912 140 3111 2508 3013 2564 2799 307 2896 2513 3032 2765 2802 221 2872 2595 2781 2578 2707 144 2476 2380 2820 2651 2582 194 2685 2602 2998 2616 2725 186 2488 2275 2807 2394 2491 228 2467 2477 2587 2067 2399 228 2183 2398 2543 2186 2328 176 1944 2326 2532 1776 2145 346 1948 2041 2791 2344 2281 380 2018 2013 2171 1721 1981 188 1832 1903 2347 2267 2087 257 1932 2169 2213 1916 2057 155 1855 1854 2316 1905 1982 224 2156 1965 2756 2343 2305 338 2564 2510 2797 2630 2625 125 3132 2821 3506 2895 3088 308 2833 2869 3164 2960 2956 149 2774 2421 3060 2920 2794 275 2464 2451 2963 2853 2683 264 Page A44 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 4: HF + 0.006 % PFHS VCO2 (ml/h/kg mouse) Average VCO2 SD cage 0103 cage 0107 cage 0203 cage 0207 (ml/h/kg mouse) 3309 3390 3535 3348 3395 99 3243 3195 3526 3375 3335 148 2910 3285 3487 2961 3161 274 3013 2980 3360 2695 3012 273 2848 2670 3385 2981 2971 304 2803 3043 3222 3000 3017 172 2757 2340 2802 2483 2596 221 2834 2384 2957 2469 2661 278 2699 2685 2527 2316 2557 179 2641 2677 2628 2448 2598 102 2474 2658 2505 2519 2539 81 2501 3011 2615 2533 2665 236 2834 2835 2489 2379 2634 236 2638 2685 2634 2873 2708 113 2739 2772 2653 2586 2687 84 3504 3473 3331 3246 3389 121 3532 3535 3688 3585 3585 73 3236 3539 3498 3374 3412 137 3243 3256 3755 3475 3432 240 2940 3382 3493 3025 3210 269 3109 2912 3516 3001 3135 267 2575 2599 3708 2979 2965 528 2536 2804 3465 2755 2890 401 2419 1953 3136 2496 2501 487 2531 2461 2999 2664 2664 239 2613 2818 3063 2466 2740 259 2766 2612 2756 2243 2594 245 2533 2538 2510 2677 2564 76 2656 2834 2715 2186 2598 284 2367 2515 2371 2250 2376 109 2316 2481 2372 2195 2341 119 2273 2767 2861 2035 2484 395 2587 2927 2695 2742 2738 142 2771 2605 2987 2323 2672 280 3419 3205 3237 2691 3138 312 3884 3492 3775 3089 3560 355 3549 3283 4014 3281 3532 345 3537 3116 3646 3478 3444 230 3286 3411 3615 3060 3343 233 3013 2658 3123 3020 2954 204 2856 2761 2950 3005 2893 108 2482 2458 3004 2631 2644 252 2653 2516 2972 2505 2662 217 2446 2003 2852 2388 2422 348 2471 2132 2679 2317 2400 232 2444 2248 2508 2381 2395 111 2317 1959 2651 2239 2292 285 2384 2357 2281 2202 2306 82 2092 1863 2403 2467 2206 281 2089 1983 2360 2074 2127 163 2019 2148 2535 2135 2209 225 2254 2256 2476 2241 2307 113 2189 2394 2578 2122 2321 207 2792 2665 2648 2485 2648 126 3502 3130 3196 2820 3162 280 3708 2929 3755 3224 3404 397 3218 2380 3623 3083 3076 518 3192 2698 3327 2906 3031 283 Metabolic cages group 6: HF + 0.003 % PFOS Time VCO2 (ml/h/kg mouse) Average VCO2 SD (hr) cage 0104 cage 0108 cage 0204 cage 0208 (ml/h/kg mouse) 6.1 3618 3732 3629 3220 3550 226 7.4 3414 3149 3553 3346 3365 168 8.6 3077 3062 3321 2963 3106 152 9.9 3331 3294 3219 3015 3215 141 11.1 3459 3188 3324 2801 3193 284 12.4 3301 2650 2970 2656 2894 310 13.6 3344 2303 2875 2554 2769 449 14.9 3167 2783 2760 2326 2759 344 16.1 2754 2270 2407 2550 2495 207 17.4 2745 2591 2365 2139 2460 265 18.6 3329 2992 2511 2371 2801 441 19.9 3154 2288 2097 2433 2493 462 21.1 2765 2771 2441 2405 2595 200 22.4 2658 2817 2364 2466 2576 201 23.6 3248 2519 2387 2668 2705 380 24.9 3643 3328 2734 2974 3170 399 26.1 3711 3457 3256 3206 3408 230 27.4 3410 3409 3637 3476 3483 107 28.6 3586 3714 3225 3340 3466 224 29.9 3324 3143 3102 2929 3124 162 31.1 3436 3095 3183 2900 3153 223 32.4 3213 2431 3022 2895 2890 333 33.6 3040 2809 3004 2693 2887 165 34.9 3259 2784 2951 2688 2921 251 36.1 3172 2333 2737 2351 2648 396 37.4 3179 2503 2817 2633 2783 294 38.6 3080 2433 2362 2717 2648 326 39.9 2992 2068 2449 2838 2587 414 41.1 2701 2306 2399 2627 2508 186 42.4 2828 2532 2224 2310 2474 270 43.6 2695 2234 2681 2499 2527 215 44.9 2775 2233 2426 2409 2461 227 46.1 2741 2411 2500 2573 2556 140 47.4 2766 2411 2616 2534 2582 149 48.6 3454 2739 3044 2867 3026 312 49.9 3931 3266 3167 3104 3367 382 51.1 3393 3634 3407 3247 3420 160 52.4 3656 3495 3236 3014 3350 283 53.6 3109 3309 3021 3121 3140 121 54.9 3360 2940 2868 2811 2995 249 56.1 3313 2510 2896 2675 2849 348 57.4 3183 2708 2683 2661 2809 250 58.6 3210 2732 2631 2576 2787 289 59.9 3309 2775 2552 2529 2791 363 61.1 3020 2472 2453 2522 2617 270 62.4 2882 2549 2424 2522 2594 199 63.6 2636 2062 2504 2233 2359 260 64.9 2675 2302 2208 2333 2379 204 66.1 2419 2040 2197 2131 2197 161 67.4 2724 2379 2060 2004 2291 332 68.6 2533 2023 2195 1971 2180 254 69.9 2450 2344 2016 2234 2261 185 71.1 2505 2242 1969 2016 2183 246 72.4 3034 2294 2265 2426 2505 360 73.6 3260 3089 2531 2702 2895 337 74.9 3398 2900 2924 3037 3065 230 76.1 3198 3204 3044 3244 3172 88 77.4 3051 3148 2734 3064 2999 182 Page A45 of A172 Appendix to TNO-report of 3M-02 Final version Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.486 4 4 0.200 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.343 4 4 0.486 4 4 1.000 4 4 1.000 4 4 0.486 4 4 0.200 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.486 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.114 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.200 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.886 4 4 0.114 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.886 group 1 vs 4 nn p 4 4 0.343 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.200 4 4 0.200 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.889 4 4 0.343 4 4 0.200 4 4 0.057 4 4 0.200 4 4 0.114 4 4 0.686 4 4 0.029 4 4 0.057 4 4 0.114 4 4 0.114 4 4 0.200 4 4 0.489 4 4 0.343 4 4 0.686 4 4 0.057 4 4 0.343 4 4 0.886 4 4 0.486 4 4 0.486 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.343 4 4 0.686 4 4 0.686 4 4 0.057 4 4 0.114 4 4 0.057 4 4 0.029 4 4 0.486 4 4 0.200 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.686 4 4 1.000 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.200 4 4 0.886 4 4 0.114 4 4 0.886 4 4 0.200 4 4 0.486 4 4 0.200 VCO2, p-values Mann Whitney U-test group 1 vs 6 group 3 vs 4 group 3 vs 6 nn p nn p nn p 4 4 0.114 4 4 0.029 4 4 0.057 4 4 0.343 4 4 0.486 4 4 0.200 4 4 0.343 4 4 0.486 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.029 4 4 0.057 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.200 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.200 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.200 4 4 0.486 4 4 0.114 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.029 4 4 0.343 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.686 4 4 0.114 4 4 0.200 4 4 0.486 4 4 0.029 4 4 0.057 4 4 0.029 4 4 0.114 4 4 0.114 4 4 0.057 4 4 0.057 4 4 0.114 4 4 0.200 4 4 0.200 4 4 0.200 4 4 0.114 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.114 4 4 0.343 4 4 0.114 4 4 0.343 4 4 0.686 4 4 0.114 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.486 4 4 1.000 4 4 1.000 4 4 1.000 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.686 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.057 4 4 0.343 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.200 4 4 0.886 4 4 0.886 4 4 0.200 4 4 0.343 4 4 0.886 4 4 0.114 4 4 0.029 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.057 4 4 0.029 4 4 0.057 4 4 0.114 4 4 0.200 4 4 0.486 4 4 0.486 4 4 0.343 4 4 0.200 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.200 4 4 0.200 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.200 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.114 4 4 0.686 4 4 0.114 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.114 4 4 0.343 4 4 0.114 4 4 0.886 4 4 0.686 4 4 0.343 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.200 4 4 0.114 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.200 4 4 0.886 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.486 4 4 1.000 4 4 0.114 4 4 0.486 4 4 0.057 4 4 0.343 4 4 0.057 4 4 0.114 4 4 0.200 4 4 0.114 group 4 vs 6 nn p 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.486 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.343 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.686 4 4 1.000 4 4 0.686 4 4 1.000 4 4 1.000 4 4 0.686 4 4 0.200 4 4 0.886 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.200 4 4 1.000 4 4 0.114 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.343 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.486 4 4 0.200 4 4 0.343 4 4 0.114 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.343 4 4 0.200 4 4 1.000 4 4 1.000 04-12-2006 p<0.05 Page A46 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Respiratory exchange ratio Metabolic cages group 1: HF control Respiratory exchange ratio cage 0101 cage 0105 cage 0201 cage 0205 0.960 0.861 1.015 0.895 1.025 0.902 1.037 0.934 0.997 0.900 0.994 0.931 0.971 0.854 0.962 0.909 1.014 0.848 0.906 0.884 0.966 0.933 0.975 0.885 0.939 0.841 1.000 0.892 0.981 0.839 0.976 0.883 1.008 0.890 0.926 0.845 0.962 0.817 0.917 0.837 0.969 0.824 0.905 0.815 0.896 0.827 0.936 0.792 0.906 0.821 0.895 0.803 0.877 0.824 0.966 0.843 0.852 0.876 0.867 0.862 0.908 0.878 0.890 0.857 0.941 0.886 0.960 0.785 0.923 0.877 0.936 0.853 0.965 0.823 1.007 0.877 0.963 0.802 0.995 0.866 0.961 0.784 0.977 0.903 0.926 0.805 0.952 0.856 0.948 0.765 0.876 0.747 0.889 0.809 0.881 0.797 0.879 0.780 0.907 0.870 0.882 0.764 0.894 0.821 0.985 0.920 0.998 0.865 1.030 0.936 1.040 0.881 1.027 0.916 1.012 0.856 0.972 0.882 0.947 0.827 0.962 0.890 0.946 0.814 0.930 0.892 0.985 0.863 0.940 0.882 0.959 0.845 0.905 0.920 0.941 0.908 0.913 0.916 0.968 0.915 0.881 0.886 0.881 0.856 0.907 0.888 0.964 0.901 0.919 0.870 0.981 0.910 0.892 0.870 0.923 0.871 0.832 0.827 0.975 0.812 0.907 0.816 0.943 0.828 0.916 0.865 0.956 0.813 0.910 0.831 0.888 0.781 0.903 0.817 0.899 0.787 0.964 0.763 0.931 0.802 0.963 0.861 0.875 0.766 0.896 0.774 0.830 0.772 0.933 0.784 0.817 0.759 0.890 0.745 0.810 0.783 0.876 0.794 0.857 0.721 0.811 0.758 0.907 0.772 0.777 0.839 0.926 0.785 0.852 0.775 0.903 0.830 0.887 0.901 0.912 0.814 0.855 0.909 0.890 0.831 0.891 0.921 0.931 0.854 0.870 0.873 0.930 0.866 0.888 0.844 0.872 0.864 Average RER 0.933 0.974 0.955 0.924 0.913 0.940 0.918 0.920 0.917 0.883 0.878 0.863 0.856 0.878 0.864 0.883 0.893 0.897 0.918 0.906 0.906 0.885 0.834 0.844 0.859 0.840 0.942 0.972 0.953 0.907 0.903 0.917 0.906 0.918 0.928 0.876 0.915 0.920 0.889 0.861 0.874 0.887 0.853 0.851 0.865 0.867 0.818 0.823 0.807 0.812 0.812 0.832 0.840 0.878 0.871 0.899 0.885 0.867 SD 0.069 0.067 0.048 0.054 0.072 0.041 0.068 0.070 0.069 0.068 0.073 0.065 0.052 0.063 0.010 0.022 0.079 0.039 0.083 0.089 0.087 0.067 0.095 0.047 0.055 0.060 0.062 0.076 0.081 0.066 0.067 0.053 0.052 0.016 0.027 0.013 0.033 0.046 0.025 0.076 0.062 0.062 0.058 0.059 0.097 0.080 0.059 0.077 0.062 0.070 0.067 0.069 0.053 0.044 0.035 0.035 0.030 0.018 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 3: HF + 0.03 % PFBS Respiratory exchange ratio Average cage 0102 cage 0106 cage 0202 cage 0206 RER 0.937 0.894 0.893 0.878 0.900 0.950 0.919 0.987 0.903 0.940 0.959 0.882 0.972 0.887 0.925 0.931 0.884 0.959 0.856 0.908 0.944 0.910 0.909 0.842 0.901 0.922 0.889 0.932 0.869 0.903 0.920 0.800 0.928 0.895 0.886 0.923 0.867 0.867 0.805 0.866 0.946 0.831 0.908 0.851 0.884 0.927 0.808 0.874 0.841 0.862 0.836 0.832 0.863 0.807 0.834 0.831 0.804 0.916 0.803 0.839 0.855 0.863 0.921 0.763 0.851 0.833 0.858 0.847 0.780 0.830 0.858 0.846 0.874 0.849 0.857 0.834 0.878 0.913 0.871 0.874 0.894 0.812 0.895 0.818 0.855 0.901 0.856 0.943 0.875 0.894 0.887 0.809 0.982 0.893 0.892 0.858 0.771 0.975 0.855 0.865 0.851 0.743 0.896 0.855 0.837 0.877 0.783 0.862 0.859 0.845 0.871 0.744 0.867 0.823 0.826 0.850 0.779 0.842 0.830 0.825 0.837 0.802 0.835 0.801 0.819 0.922 0.765 0.852 0.871 0.852 0.994 0.868 0.960 0.929 0.938 0.988 0.870 0.957 0.917 0.933 0.998 0.920 0.955 0.904 0.944 0.949 0.901 0.864 0.811 0.881 0.916 0.931 0.829 0.766 0.860 0.912 0.842 0.825 0.824 0.851 0.912 0.851 0.833 0.821 0.854 0.906 0.883 0.900 0.893 0.895 0.854 0.919 0.888 0.861 0.881 0.890 0.902 0.902 0.809 0.876 0.929 0.892 0.882 0.845 0.887 0.958 0.907 0.892 0.893 0.912 0.928 0.895 0.873 0.808 0.876 0.938 0.875 0.853 0.840 0.877 0.930 0.855 0.814 0.809 0.852 0.864 0.826 0.829 0.812 0.833 0.870 0.829 0.832 0.790 0.830 0.815 0.800 0.811 0.783 0.802 0.922 0.786 0.840 0.784 0.833 0.884 0.858 0.880 0.803 0.856 0.809 0.860 0.861 0.746 0.819 0.833 0.854 0.863 0.801 0.838 0.793 0.877 0.770 0.719 0.790 0.759 0.822 0.778 0.790 0.787 0.783 0.814 0.789 0.760 0.787 0.754 0.758 0.811 0.767 0.772 0.794 0.844 0.841 0.835 0.828 0.787 0.873 0.794 0.830 0.821 0.894 0.867 0.862 0.809 0.858 0.879 0.856 0.875 0.854 0.866 0.892 0.826 0.893 0.850 0.866 0.821 0.826 0.826 0.853 0.831 SD 0.025 0.037 0.047 0.046 0.042 0.029 0.059 0.048 0.053 0.051 0.023 0.053 0.065 0.034 0.013 0.032 0.046 0.037 0.071 0.084 0.065 0.042 0.059 0.032 0.020 0.065 0.053 0.051 0.042 0.059 0.077 0.041 0.040 0.010 0.030 0.045 0.034 0.031 0.051 0.044 0.056 0.022 0.033 0.014 0.065 0.037 0.054 0.027 0.066 0.027 0.022 0.026 0.023 0.039 0.036 0.013 0.033 0.014 Page A47 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 4: HF + 0.006 % PFHS Respiratory exchange ratio Average cage 0103 cage 0107 cage 0203 cage 0207 RER 0.927 0.918 0.932 0.913 0.922 0.944 0.925 0.942 0.944 0.939 0.889 0.920 0.938 0.922 0.917 0.913 0.905 0.907 0.894 0.905 0.885 0.881 0.896 0.920 0.895 0.858 0.889 0.876 0.895 0.880 0.848 0.843 0.869 0.852 0.853 0.901 0.817 0.861 0.860 0.860 0.862 0.791 0.795 0.802 0.812 0.851 0.807 0.823 0.884 0.841 0.820 0.918 0.843 0.837 0.854 0.810 0.952 0.803 0.884 0.862 0.919 0.943 0.842 0.839 0.886 0.863 0.905 0.824 0.857 0.862 0.845 0.880 0.802 0.845 0.843 0.868 0.787 0.879 0.888 0.855 0.829 0.828 0.875 0.871 0.851 0.855 0.912 0.912 0.899 0.894 0.922 0.968 0.948 0.926 0.941 0.933 0.959 0.960 0.934 0.947 0.915 0.919 0.963 0.932 0.932 0.815 0.912 0.930 0.916 0.893 0.795 0.872 0.882 0.833 0.846 0.813 0.806 0.870 0.812 0.825 0.794 0.851 0.896 0.801 0.836 0.866 0.938 0.866 0.749 0.855 0.942 0.939 0.856 0.845 0.895 0.890 0.945 0.812 0.929 0.894 0.903 0.968 0.843 0.867 0.895 0.812 0.919 0.795 0.821 0.837 0.781 0.925 0.796 0.794 0.824 0.813 0.923 0.829 0.754 0.830 0.840 0.892 0.884 0.857 0.868 0.860 0.910 0.915 0.861 0.887 0.925 0.913 0.924 0.871 0.908 0.922 0.882 0.921 0.839 0.891 0.939 0.890 0.922 0.885 0.909 0.964 0.919 0.931 0.926 0.935 0.945 0.958 0.940 0.922 0.941 0.900 0.909 0.840 0.908 0.889 0.863 0.898 0.830 0.893 0.871 0.819 0.819 0.803 0.862 0.826 0.814 0.801 0.803 0.792 0.803 0.823 0.777 0.829 0.810 0.810 0.820 0.803 0.828 0.839 0.822 0.809 0.786 0.834 0.849 0.819 0.801 0.777 0.795 0.838 0.803 0.817 0.811 0.745 0.832 0.801 0.802 0.781 0.753 0.806 0.786 0.754 0.798 0.785 0.771 0.777 0.754 0.786 0.820 0.796 0.789 0.789 0.769 0.819 0.803 0.795 0.763 0.802 0.846 0.809 0.805 0.862 0.809 0.804 0.850 0.831 0.924 0.781 0.841 0.822 0.842 0.939 0.789 0.877 0.846 0.863 0.955 0.799 0.878 0.888 0.880 0.936 0.836 0.841 0.860 0.868 SD 0.008 0.009 0.020 0.008 0.018 0.016 0.012 0.034 0.034 0.034 0.044 0.070 0.053 0.033 0.032 0.046 0.026 0.027 0.021 0.015 0.022 0.053 0.040 0.030 0.048 0.078 0.052 0.059 0.055 0.056 0.068 0.070 0.024 0.030 0.025 0.040 0.026 0.020 0.015 0.033 0.031 0.025 0.009 0.023 0.015 0.028 0.026 0.039 0.025 0.019 0.027 0.021 0.034 0.029 0.060 0.063 0.064 0.047 Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 Metabolic cages group 6: HF + 0.003 % PFOS Respiratory exchange ratio Average cage 0104 cage 0108 cage 0204 cage 0208 RER 0.920 0.863 0.914 0.888 0.896 0.903 0.790 0.935 0.916 0.886 0.923 0.802 0.903 0.878 0.876 0.900 0.871 0.899 0.877 0.887 0.894 0.874 0.911 0.891 0.892 0.909 0.821 0.887 0.850 0.867 0.888 0.741 0.893 0.799 0.830 0.882 0.796 0.848 0.774 0.825 0.826 0.782 0.777 0.839 0.806 0.855 0.817 0.787 0.771 0.808 0.891 0.850 0.782 0.814 0.834 0.853 0.772 0.718 0.805 0.787 0.835 0.895 0.828 0.785 0.836 0.806 0.845 0.793 0.818 0.815 0.868 0.784 0.771 0.830 0.813 0.892 0.828 0.814 0.875 0.852 0.851 0.871 0.804 0.848 0.844 0.908 0.881 0.879 0.886 0.888 0.911 0.914 0.894 0.885 0.901 0.886 0.900 0.909 0.848 0.885 0.868 0.840 0.912 0.862 0.871 0.843 0.764 0.888 0.827 0.830 0.776 0.819 0.836 0.812 0.811 0.836 0.812 0.865 0.830 0.836 0.824 0.756 0.828 0.761 0.792 0.890 0.825 0.847 0.839 0.850 0.929 0.807 0.801 0.869 0.852 0.916 0.759 0.828 0.902 0.851 0.867 0.813 0.807 0.871 0.839 0.898 0.784 0.793 0.790 0.816 0.808 0.787 0.846 0.782 0.806 0.816 0.785 0.830 0.825 0.814 0.827 0.820 0.804 0.834 0.821 0.836 0.826 0.885 0.822 0.842 0.896 0.827 0.859 0.883 0.866 0.890 0.864 0.860 0.857 0.868 0.885 0.909 0.891 0.836 0.880 0.905 0.901 0.897 0.837 0.885 0.829 0.885 0.860 0.861 0.859 0.860 0.852 0.813 0.841 0.841 0.835 0.792 0.799 0.784 0.802 0.852 0.795 0.743 0.768 0.790 0.846 0.817 0.779 0.750 0.798 0.855 0.834 0.735 0.745 0.792 0.841 0.783 0.762 0.784 0.793 0.782 0.803 0.806 0.795 0.797 0.838 0.741 0.813 0.772 0.791 0.833 0.776 0.816 0.763 0.797 0.820 0.807 0.753 0.749 0.782 0.833 0.811 0.715 0.736 0.774 0.787 0.758 0.766 0.709 0.755 0.773 0.792 0.757 0.733 0.764 0.809 0.782 0.715 0.722 0.757 0.850 0.786 0.755 0.775 0.792 0.868 0.838 0.788 0.825 0.830 0.885 0.871 0.835 0.825 0.854 0.876 0.893 0.878 0.846 0.873 0.814 0.869 0.811 0.832 0.832 SD 0.026 0.066 0.053 0.015 0.015 0.039 0.074 0.049 0.031 0.037 0.047 0.057 0.045 0.022 0.044 0.037 0.028 0.013 0.014 0.027 0.030 0.051 0.025 0.022 0.039 0.028 0.060 0.073 0.034 0.055 0.029 0.020 0.013 0.029 0.030 0.015 0.031 0.032 0.023 0.021 0.023 0.047 0.042 0.061 0.034 0.011 0.043 0.033 0.037 0.057 0.033 0.025 0.046 0.041 0.033 0.028 0.020 0.027 Page A48 of A172 Appendix to TNO-report of 3M-02 Final version Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.486 4 4 0.686 4 4 0.343 4 4 0.686 4 4 0.886 4 4 0.200 4 4 0.686 4 4 0.343 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.486 4 4 0.343 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.886 4 4 1.000 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.114 4 4 0.200 4 4 0.057 4 4 0.200 4 4 0.343 4 4 0.343 4 4 0.686 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.686 4 4 1.000 4 4 1.000 4 4 0.486 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.114 4 4 0.886 4 4 0.114 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.057 Respiratory exchange ratio, p-values Mann Whitney U-test group 1 vs 4 group 1 vs 6 group 3 vs 4 group 3 vs 6 group 4 vs 6 nn p nn p nn p nn p nn p 4 4 1.000 4 4 0.686 4 4 0.343 4 4 0.886 4 4 0.200 4 4 0.886 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.057 4 4 0.343 4 4 0.114 4 4 1.000 4 4 0.343 4 4 0.343 4 4 0.486 4 4 0.343 4 4 1.000 4 4 0.686 4 4 0.114 4 4 1.000 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.114 4 4 0.114 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.114 4 4 1.000 4 4 0.343 4 4 0.114 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.057 4 4 0.029 4 4 0.114 4 4 0.057 4 4 0.686 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.486 4 4 1.000 4 4 0.686 4 4 1.000 4 4 0.200 4 4 0.886 4 4 0.486 4 4 0.200 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.686 4 4 0.114 4 4 0.886 4 4 0.114 4 4 0.343 4 4 0.486 4 4 0.057 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.486 4 4 0.343 4 4 0.886 4 4 0.486 4 4 1.000 4 4 0.343 4 4 0.343 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.886 4 4 1.000 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.886 4 4 1.000 4 4 0.343 4 4 0.686 4 4 0.029 4 4 1.000 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.029 4 4 0.886 4 4 0.486 4 4 0.029 4 4 0.486 4 4 0.029 4 4 1.000 4 4 0.343 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.886 4 4 1.000 4 4 0.486 4 4 0.486 4 4 0.343 4 4 0.886 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.486 4 4 0.686 4 4 0.114 4 4 0.886 4 4 0.343 4 4 0.343 4 4 1.000 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.343 4 4 0.114 4 4 0.486 4 4 0.114 4 4 0.486 4 4 0.343 4 4 0.114 4 4 0.200 4 4 0.029 4 4 0.343 4 4 0.114 4 4 0.114 4 4 0.486 4 4 0.114 4 4 0.200 4 4 0.114 4 4 0.057 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.114 4 4 0.029 4 4 0.686 4 4 0.343 4 4 1.000 4 4 0.486 4 4 0.029 4 4 0.486 4 4 0.200 4 4 0.029 4 4 0.343 4 4 0.029 4 4 1.000 4 4 0.057 4 4 0.114 4 4 0.686 4 4 0.029 4 4 0.200 4 4 0.686 4 4 0.114 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.886 4 4 0.343 4 4 0.343 4 4 0.200 4 4 0.200 4 4 0.686 4 4 0.029 4 4 0.057 4 4 0.114 4 4 0.057 4 4 0.486 4 4 0.029 4 4 0.343 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.200 4 4 1.000 4 4 0.114 4 4 0.486 4 4 0.114 4 4 0.057 4 4 0.200 4 4 0.057 4 4 0.486 4 4 0.200 4 4 0.200 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.343 4 4 1.000 4 4 0.486 4 4 0.343 4 4 0.686 4 4 1.000 4 4 1.000 4 4 1.000 4 4 0.343 4 4 1.000 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.343 4 4 0.200 4 4 0.114 4 4 0.200 4 4 0.886 4 4 0.686 4 4 0.486 4 4 0.486 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.200 4 4 0.114 4 4 1.000 4 4 0.686 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.486 4 4 0.486 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.200 4 4 0.886 4 4 0.200 4 4 0.343 4 4 0.686 4 4 0.114 4 4 0.200 4 4 0.886 4 4 0.200 4 4 0.343 4 4 0.114 4 4 0.686 4 4 0.057 4 4 0.200 4 4 0.114 4 4 0.057 4 4 0.686 4 4 0.200 4 4 0.200 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.486 4 4 1.000 4 4 0.486 4 4 0.114 4 4 0.886 4 4 0.686 4 4 0.886 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.114 4 4 0.114 4 4 0.886 4 4 0.200 04-12-2006 p<0.05 Page A49 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 D6. Energy expenditure Metabolic cages group 1: HF control Time Energy expenditure (kcal/h/kg mouse) Average energy expenditure SD (hr) cage 0101 cage 0105 cage 0201 cage 0205 (kcal/h/kg mouse) 6.1 15.26 17.60 18.09 15.65 16.65 1.40 7.4 14.77 18.58 16.69 14.83 16.22 1.81 8.6 14.06 17.22 15.46 15.02 15.44 1.32 9.9 15.29 17.23 18.03 14.38 16.23 1.69 11.1 13.87 15.56 14.51 15.73 14.92 0.88 12.4 14.09 14.71 16.01 14.24 14.76 0.88 13.6 13.11 15.06 13.84 13.08 13.77 0.93 14.9 13.48 13.36 13.89 13.27 13.50 0.27 16.1 12.13 13.82 13.73 13.00 13.17 0.78 17.4 14.07 13.01 14.23 13.37 13.67 0.58 18.6 15.59 13.78 12.86 13.16 13.85 1.22 19.9 13.62 14.11 14.61 14.87 14.30 0.56 21.1 12.84 14.44 12.93 12.21 13.11 0.95 22.4 13.44 13.08 14.15 14.04 13.68 0.51 23.6 14.61 14.60 14.57 13.58 14.34 0.50 24.9 18.76 18.08 18.68 19.18 18.67 0.45 26.1 14.71 16.46 17.67 18.47 16.83 1.64 27.4 16.80 17.25 17.38 14.72 16.54 1.24 28.6 14.60 15.94 17.12 17.36 16.25 1.27 29.9 14.85 15.69 15.70 14.61 15.21 0.57 31.1 14.90 15.61 16.65 14.99 15.54 0.80 32.4 13.70 15.30 16.48 15.57 15.26 1.16 33.6 13.66 15.14 16.64 13.85 14.82 1.38 34.9 15.51 14.87 15.70 13.65 14.93 0.93 36.1 13.93 14.80 13.31 13.10 13.78 0.76 37.4 13.90 14.00 16.10 13.12 14.28 1.28 38.6 13.43 12.69 14.25 12.34 13.18 0.84 39.9 12.20 13.39 14.13 13.03 13.19 0.80 41.1 13.65 13.20 13.31 11.93 13.02 0.75 42.4 11.75 13.71 13.91 13.69 13.27 1.01 43.6 12.71 12.39 13.16 12.74 12.75 0.32 44.9 12.64 13.18 14.44 12.48 13.19 0.89 46.1 13.72 14.15 14.99 13.38 14.06 0.70 47.4 14.48 14.15 13.45 13.61 13.93 0.48 48.6 17.69 17.46 18.78 16.77 17.67 0.84 49.9 16.95 16.08 18.41 16.32 16.94 1.05 51.1 16.37 17.06 17.66 15.18 16.57 1.06 52.4 14.77 16.94 16.72 15.37 15.95 1.05 53.6 14.41 15.64 15.92 15.35 15.33 0.66 54.9 14.84 15.90 16.22 14.90 15.46 0.70 56.1 13.10 15.64 15.87 13.62 14.56 1.40 57.4 13.93 15.75 16.27 14.08 15.01 1.18 58.6 13.93 13.92 16.08 14.73 14.67 1.02 59.9 14.30 15.21 15.35 12.69 14.39 1.22 61.1 13.41 13.41 13.86 12.72 13.35 0.47 62.4 11.94 14.05 13.54 11.28 12.70 1.31 63.6 12.07 12.94 14.22 12.19 12.86 0.99 64.9 13.53 12.28 14.41 11.88 13.02 1.16 66.1 11.78 10.77 11.51 12.25 11.58 0.62 67.4 12.67 12.95 13.74 11.51 12.72 0.92 68.6 11.52 12.61 12.64 12.73 12.37 0.57 69.9 12.09 12.89 12.94 11.99 12.48 0.51 71.1 12.71 11.98 13.88 13.18 12.94 0.80 72.4 15.29 15.54 16.61 15.93 15.84 0.57 73.6 16.15 17.21 19.20 16.67 17.31 1.33 74.9 16.12 16.43 17.30 16.02 16.47 0.58 76.1 14.28 15.90 16.75 14.24 15.29 1.24 77.4 14.01 16.21 16.58 15.16 15.49 1.16 Metabolic cages group 3: HF + 0.03 % PFBS Time Energy expenditure (kcal/h/kg mouse) Average energy expenditure SD (hr) cage 0102 cage 0106 cage 0202 cage 0206 (kcal/h/kg mouse) 6.1 17.43 16.55 15.54 17.20 16.68 0.85 7.4 17.09 16.86 16.94 17.80 17.17 0.43 8.6 15.39 15.70 17.29 16.16 16.13 0.83 9.9 15.91 14.45 15.57 16.99 15.73 1.05 11.1 16.34 15.43 18.84 15.42 16.51 1.61 12.4 14.64 15.03 16.85 14.16 15.17 1.17 13.6 15.19 12.93 15.35 15.09 14.64 1.14 14.9 13.95 13.93 15.14 12.27 13.82 1.18 16.1 14.44 13.92 16.41 12.52 14.32 1.61 17.4 14.01 12.76 17.22 13.38 14.34 1.99 18.6 14.98 14.58 15.00 12.81 14.34 1.04 19.9 12.72 13.02 15.40 12.07 13.30 1.45 21.1 13.81 13.65 16.90 13.71 14.52 1.59 22.4 13.46 14.25 15.08 13.72 14.13 0.72 23.6 15.62 12.57 17.30 13.56 14.76 2.12 24.9 18.75 18.37 20.52 17.56 18.80 1.25 26.1 18.06 19.16 19.87 18.43 18.88 0.80 27.4 16.48 15.80 17.65 17.88 16.95 0.98 28.6 16.92 12.95 16.88 15.09 15.46 1.88 29.9 15.90 16.54 17.00 16.45 16.47 0.45 31.1 14.09 12.21 17.31 15.23 14.71 2.13 32.4 14.12 13.77 17.36 14.77 15.00 1.62 33.6 14.78 12.74 16.24 13.97 14.43 1.47 34.9 15.12 13.05 16.89 15.06 15.03 1.57 36.1 14.52 14.64 16.12 13.22 14.62 1.19 37.4 15.83 12.02 16.36 14.18 14.60 1.95 38.6 13.76 12.26 15.82 12.58 13.61 1.61 39.9 13.81 12.37 15.88 12.21 13.57 1.70 41.1 13.39 12.95 15.06 12.52 13.48 1.11 42.4 13.12 10.84 16.01 15.10 13.77 2.29 43.6 13.64 11.69 15.73 13.03 13.52 1.68 44.9 13.95 12.08 16.21 13.22 13.86 1.74 46.1 13.47 12.78 15.51 12.95 13.68 1.25 47.4 15.18 12.38 16.46 13.52 14.39 1.79 48.6 17.57 15.45 18.91 17.72 17.41 1.44 49.9 19.09 17.24 18.98 18.82 18.53 0.87 51.1 17.30 16.92 17.99 17.07 17.32 0.48 52.4 16.05 14.75 16.27 16.18 15.81 0.71 53.6 16.77 13.77 16.86 15.23 15.66 1.46 54.9 15.50 14.08 17.30 15.96 15.71 1.33 56.1 15.48 14.73 16.46 15.33 15.50 0.72 57.4 14.11 13.92 16.45 15.71 15.05 1.23 58.6 15.24 15.13 17.39 15.86 15.91 1.04 59.9 14.87 13.68 16.69 14.54 14.94 1.27 61.1 13.37 15.05 14.93 12.60 13.99 1.20 62.4 12.14 13.59 14.17 13.05 13.24 0.86 63.6 11.67 13.16 14.32 11.27 12.60 1.40 64.9 11.45 11.64 15.69 13.95 13.18 2.02 66.1 12.32 11.23 13.41 11.25 12.05 1.04 67.4 11.54 11.21 14.36 13.65 12.69 1.55 68.6 11.90 12.82 13.42 11.92 12.52 0.74 69.9 11.77 11.55 13.71 11.82 12.21 1.00 71.1 13.05 11.30 15.86 13.56 13.44 1.88 72.4 15.71 14.02 16.76 15.30 15.44 1.13 73.6 17.40 15.88 19.87 17.20 17.59 1.67 74.9 15.92 16.23 17.67 16.87 16.68 0.77 76.1 15.41 14.16 16.82 16.69 15.77 1.25 77.4 14.65 14.28 17.27 16.29 15.62 1.40 Page A50 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Metabolic cages group 4: HF + 0.006 % PFHS Time Energy expenditure (kcal/h/kg mouse) Average energy expenditure SD (hr) cage 0103 cage 0107 cage 0203 cage 0207 (kcal/h/kg mouse) 6.1 17.72 18.32 18.90 18.10 18.26 0.49 7.4 17.11 17.10 18.69 17.78 17.67 0.75 8.6 16.08 17.64 18.56 15.91 17.05 1.27 9.9 16.29 16.21 18.34 14.84 16.42 1.44 11.1 15.79 14.83 18.65 16.03 16.33 1.64 12.4 15.90 16.76 18.05 16.46 16.79 0.91 13.6 15.81 13.50 15.81 14.17 14.82 1.17 14.9 15.50 14.04 16.75 14.00 15.07 1.32 16.1 15.27 16.11 15.26 13.82 15.12 0.95 17.4 15.08 15.83 15.47 13.59 14.99 0.99 18.6 14.56 14.33 14.50 14.54 14.48 0.11 19.9 14.82 15.78 15.58 14.07 15.06 0.78 21.1 15.26 14.93 14.36 13.74 14.57 0.67 22.4 14.90 14.62 15.33 16.29 15.28 0.73 23.6 15.70 15.43 15.87 14.86 15.46 0.44 24.9 19.72 21.15 18.58 17.89 19.33 1.43 26.1 20.61 20.63 20.66 20.08 20.50 0.28 27.4 18.46 19.15 18.98 18.45 18.76 0.36 28.6 17.40 16.86 19.80 18.59 18.16 1.31 29.9 15.63 17.63 18.26 16.07 16.90 1.25 31.1 16.78 15.69 18.33 15.98 16.69 1.18 32.4 15.19 14.09 19.84 16.07 16.29 2.50 33.6 15.21 15.71 19.30 15.97 16.55 1.86 34.9 14.31 11.67 17.68 14.78 14.61 2.46 36.1 15.18 14.05 16.54 15.91 15.42 1.07 37.4 14.73 15.03 17.36 15.56 15.67 1.18 38.6 14.63 13.86 15.71 12.89 14.27 1.20 39.9 13.99 13.36 14.92 14.30 14.14 0.65 41.1 14.49 14.67 15.65 12.34 14.29 1.39 42.4 14.01 13.51 14.35 13.17 13.76 0.52 43.6 14.08 13.28 14.29 13.21 13.71 0.55 44.9 13.47 14.84 16.71 12.78 14.45 1.73 46.1 14.88 16.08 15.00 15.51 15.37 0.55 47.4 15.64 14.14 16.17 13.12 14.77 1.40 48.6 18.28 17.34 17.39 15.11 17.03 1.35 49.9 20.86 19.42 20.34 17.84 19.62 1.33 51.1 18.77 18.11 21.60 18.16 19.16 1.65 52.4 18.37 16.76 19.49 18.59 18.30 1.14 53.6 17.30 17.80 19.21 16.43 17.68 1.16 54.9 16.46 14.44 18.06 16.42 16.35 1.48 56.1 16.08 15.13 17.28 16.52 16.25 0.90 57.4 14.64 14.47 18.02 14.88 15.50 1.69 58.6 15.67 15.08 17.87 15.07 15.92 1.33 59.9 14.27 12.31 16.74 14.16 14.37 1.82 61.1 14.48 12.77 15.72 13.40 14.09 1.30 62.4 14.56 13.63 14.62 13.60 14.11 0.56 63.6 13.88 12.02 15.96 12.96 13.70 1.69 64.9 13.99 13.95 14.50 12.77 13.80 0.73 66.1 12.49 11.40 15.11 14.61 13.40 1.75 67.4 13.11 11.93 14.36 12.75 13.04 1.01 68.6 12.66 12.96 14.99 12.77 13.35 1.10 69.9 13.64 13.88 14.56 13.32 13.85 0.53 71.1 13.62 14.26 14.77 12.59 13.81 0.94 72.4 15.79 15.84 15.90 14.20 15.43 0.82 73.6 18.77 19.12 18.49 16.54 18.23 1.16 74.9 19.64 17.74 21.04 18.51 19.23 1.43 76.1 16.81 14.27 20.24 17.03 17.09 2.45 77.4 16.94 15.58 19.19 16.40 17.03 1.55 Metabolic cages group 6: HF + 0.003 % PFOS Time Energy expenditure (kcal/h/kg mouse) Average energy expenditure SD (hr) cage 0104 cage 0108 cage 0204 cage 0208 (kcal/h/kg mouse) 6.1 19.47 21.08 19.61 17.81 19.49 1.34 7.4 18.62 19.06 18.86 18.04 18.65 0.44 8.6 16.51 18.30 18.11 16.54 17.36 0.97 9.9 18.20 18.49 17.62 16.84 17.79 0.73 11.1 19.01 17.84 18.00 15.43 17.57 1.52 12.4 17.93 15.57 16.43 15.19 16.28 1.22 13.6 18.49 14.69 15.83 15.23 16.06 1.68 14.9 17.61 16.67 15.79 14.34 16.10 1.39 16.1 16.12 13.87 14.75 14.73 14.87 0.93 17.4 15.64 15.29 14.35 13.23 14.63 1.08 18.6 18.32 17.05 15.27 13.96 16.15 1.92 19.9 17.91 14.12 13.69 14.47 15.05 1.94 21.1 16.01 15.24 14.26 14.58 15.02 0.78 22.4 15.85 16.13 14.28 14.54 15.20 0.93 23.6 18.23 15.33 14.68 15.49 15.93 1.57 24.9 20.12 19.45 16.14 16.60 18.08 2.00 26.1 21.17 19.41 19.46 18.32 19.59 1.18 27.4 18.52 18.96 20.24 19.23 19.24 0.73 28.6 19.45 20.08 17.71 18.51 18.94 1.04 29.9 18.39 17.21 16.81 16.76 17.29 0.76 31.1 19.29 17.83 17.23 16.41 17.69 1.22 32.4 18.53 15.12 16.72 16.89 16.81 1.39 33.6 18.69 16.53 17.36 15.99 17.14 1.18 34.9 18.84 16.51 16.59 15.65 16.90 1.36 36.1 18.59 14.60 15.98 14.62 15.95 1.88 37.4 17.62 14.68 16.18 15.30 15.94 1.27 38.6 16.44 14.48 14.14 15.27 15.08 1.02 39.9 16.15 12.94 14.29 15.48 14.72 1.41 41.1 15.21 13.64 14.29 14.75 14.47 0.67 42.4 15.50 15.37 13.42 14.02 14.58 1.02 43.6 15.94 13.52 15.32 15.22 15.00 1.04 44.9 16.36 13.59 14.18 14.09 14.56 1.23 46.1 16.01 14.15 14.92 14.94 15.00 0.76 47.4 16.00 14.11 14.52 14.85 14.87 0.81 48.6 18.97 15.99 17.26 15.91 17.03 1.43 49.9 21.66 18.42 17.93 17.63 18.91 1.86 51.1 18.81 19.72 18.76 18.78 19.02 0.47 52.4 19.91 19.11 17.76 17.42 18.55 1.16 53.6 18.13 18.31 17.13 17.61 17.79 0.53 54.9 19.04 16.79 16.98 16.20 17.25 1.24 56.1 19.21 15.17 17.38 16.28 17.01 1.72 57.4 18.16 16.35 17.05 16.49 17.01 0.82 58.6 18.42 16.12 16.12 16.24 16.73 1.13 59.9 18.84 16.11 16.36 16.05 16.84 1.34 61.1 17.44 15.08 15.28 15.33 15.78 1.11 62.4 17.66 15.22 14.46 15.22 15.64 1.39 63.6 15.27 13.11 14.82 13.76 14.24 0.98 64.9 15.52 14.03 13.04 14.49 14.27 1.03 66.1 14.25 12.15 13.77 13.48 13.41 0.90 67.4 15.72 14.09 13.50 12.84 14.04 1.23 68.6 15.34 12.66 13.55 13.02 13.64 1.19 69.9 15.08 14.14 12.63 14.25 14.02 1.02 71.1 14.87 13.67 12.91 13.09 13.64 0.89 72.4 17.36 13.95 14.19 14.87 15.09 1.56 73.6 18.36 17.87 15.30 15.80 16.83 1.51 74.9 18.82 16.27 16.93 17.77 17.45 1.10 76.1 17.86 17.62 16.97 18.62 17.76 0.68 77.4 18.01 17.66 16.20 17.84 17.43 0.83 Page A51 of A172 Appendix to TNO-report of 3M-02 Final version Time (hr) 6.1 7.4 8.6 9.9 11.1 12.4 13.6 14.9 16.1 17.4 18.6 19.9 21.1 22.4 23.6 24.9 26.1 27.4 28.6 29.9 31.1 32.4 33.6 34.9 36.1 37.4 38.6 39.9 41.1 42.4 43.6 44.9 46.1 47.4 48.6 49.9 51.1 52.4 53.6 54.9 56.1 57.4 58.6 59.9 61.1 62.4 63.6 64.9 66.1 67.4 68.6 69.9 71.1 72.4 73.6 74.9 76.1 77.4 group 1 vs 3 nn p 4 4 0.886 4 4 0.343 4 4 0.343 4 4 0.886 4 4 0.343 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.200 4 4 1.000 4 4 0.886 4 4 0.343 4 4 0.200 4 4 0.343 4 4 1.000 4 4 0.886 4 4 0.114 4 4 0.686 4 4 0.486 4 4 0.029 4 4 0.686 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.486 4 4 0.686 4 4 1.000 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.057 4 4 0.343 4 4 0.686 4 4 0.886 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.200 4 4 0.886 4 4 0.886 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.686 4 4 1.000 4 4 0.686 4 4 0.343 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.886 Energy expenditure, p-values Mann Whitney U-test group 1 vs 4 group 1 vs 6 group 3 vs 4 group 3 vs 6 group 4 vs 6 nn p nn p nn p nn p nn p 4 4 0.057 4 4 0.057 4 4 0.029 4 4 0.029 4 4 0.200 4 4 0.200 4 4 0.057 4 4 0.200 4 4 0.029 4 4 0.114 4 4 0.114 4 4 0.114 4 4 0.343 4 4 0.114 4 4 0.686 4 4 0.886 4 4 0.200 4 4 0.486 4 4 0.057 4 4 0.200 4 4 0.114 4 4 0.114 4 4 0.886 4 4 0.343 4 4 0.486 4 4 0.057 4 4 0.114 4 4 0.200 4 4 0.200 4 4 0.343 4 4 0.200 4 4 0.057 4 4 0.686 4 4 0.343 4 4 0.343 4 4 0.029 4 4 0.029 4 4 0.114 4 4 0.057 4 4 0.343 4 4 0.029 4 4 0.029 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.114 4 4 0.200 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.114 4 4 0.343 4 4 0.200 4 4 0.343 4 4 0.343 4 4 0.886 4 4 0.114 4 4 0.200 4 4 0.686 4 4 0.114 4 4 0.057 4 4 0.486 4 4 0.343 4 4 0.686 4 4 0.029 4 4 0.029 4 4 0.114 4 4 0.114 4 4 0.686 4 4 0.029 4 4 0.029 4 4 0.686 4 4 0.686 4 4 0.886 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.029 4 4 0.057 4 4 0.029 4 4 0.486 4 4 0.343 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.343 4 4 0.114 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.486 4 4 0.114 4 4 0.029 4 4 0.886 4 4 0.114 4 4 0.686 4 4 0.114 4 4 0.057 4 4 0.200 4 4 0.114 4 4 0.343 4 4 0.886 4 4 0.200 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.114 4 4 0.200 4 4 0.057 4 4 0.343 4 4 0.686 4 4 0.057 4 4 0.686 4 4 0.200 4 4 0.200 4 4 0.057 4 4 0.114 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.200 4 4 0.114 4 4 0.686 4 4 0.486 4 4 0.886 4 4 0.200 4 4 0.057 4 4 0.486 4 4 0.200 4 4 0.486 4 4 0.200 4 4 0.200 4 4 0.486 4 4 0.343 4 4 0.686 4 4 0.200 4 4 0.057 4 4 0.486 4 4 0.200 4 4 1.000 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.686 4 4 0.343 4 4 0.029 4 4 0.029 4 4 1.000 4 4 0.343 4 4 0.114 4 4 0.200 4 4 0.200 4 4 0.686 4 4 0.343 4 4 0.886 4 4 0.057 4 4 0.200 4 4 0.686 4 4 0.200 4 4 0.486 4 4 0.686 4 4 0.114 4 4 0.114 4 4 0.886 4 4 1.000 4 4 0.486 4 4 0.486 4 4 0.886 4 4 0.886 4 4 0.886 4 4 0.057 4 4 0.114 4 4 0.486 4 4 0.886 4 4 0.686 4 4 0.029 4 4 0.029 4 4 0.200 4 4 0.029 4 4 0.486 4 4 0.057 4 4 0.029 4 4 0.029 4 4 0.029 4 4 0.886 4 4 0.029 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.886 4 4 0.343 4 4 0.057 4 4 0.486 4 4 0.200 4 4 0.486 4 4 0.114 4 4 0.114 4 4 0.343 4 4 0.343 4 4 0.486 4 4 0.686 4 4 0.029 4 4 0.686 4 4 0.057 4 4 0.200 4 4 0.200 4 4 0.029 4 4 0.686 4 4 0.200 4 4 0.200 4 4 0.686 4 4 0.029 4 4 0.686 4 4 0.200 4 4 0.210 4 4 0.686 4 4 0.029 4 4 0.886 4 4 0.029 4 4 0.200 4 4 0.114 4 4 0.029 4 4 0.114 4 4 0.029 4 4 0.114 4 4 0.686 4 4 0.114 4 4 0.486 4 4 0.200 4 4 0.686 4 4 0.343 4 4 0.200 4 4 0.486 4 4 0.486 4 4 0.486 4 4 0.200 4 4 0.057 4 4 0.200 4 4 0.114 4 4 0.886 4 4 0.686 4 4 0.200 4 4 0.686 4 4 0.486 4 4 0.343 4 4 0.057 4 4 0.057 4 4 0.486 4 4 0.200 4 4 0.486 4 4 0.029 4 4 0.114 4 4 0.114 4 4 0.057 4 4 0.686 4 4 0.343 4 4 0.486 4 4 0.686 4 4 0.886 4 4 1.000 4 4 0.686 4 4 0.343 4 4 0.686 4 4 0.686 4 4 0.486 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.686 4 4 0.114 4 4 0.029 4 4 0.200 4 4 0.029 4 4 0.200 4 4 0.200 4 4 0.200 4 4 0.029 4 4 0.343 4 4 0.029 4 4 0.486 4 4 0.200 4 4 0.114 4 4 0.343 4 4 0.114 4 4 0.686 04-12-2006 p<0.05 Page A52 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Appendix E. Fecal bile acid, sterol and fatty acid measurements E1. Fecal weight measurements, bodyweight and food intake measurements Bodyweight and food intake during collecting feces Group# 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 5 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 cage# 1 1 2 2 3 3 4 5 5 5 6 6 6 7 8 8 8 9 9 10 11 12 13 13 14 15 15 15 16 17 18 18 19 19 19 19 20 20 21 21 22 22 22 22 23 24 25 25 26 27 28 29 29 30 30 31 31 31 31 32 33 34 35 36 36 37 38 38 39 39 eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 collection point 1; 2 days feces collection start collection point 1 end collection point 1 bodyweight (g) food on cage (g) bodyweight (g) food off cage (g) 33.9 33.9 31.2 49.7 30.5 38.9 30.4 31.0 28.1 51.9 28.2 39.5 31.1 30.6 29.5 47.4 29.4 35.8 26.7 26.9 27.2 20.5 30.4 30.6 30.6 31.0 27.2 61.6 27.3 44.8 33.0 33.1 29.8 30.0 31.2 68.5 32.0 48.6 32.8 31.7 33.1 24.8 30.8 31.0 30.1 29.8 32.8 68.0 32.8 47.2 26.5 26.6 26.9 42.3 26.6 31.8 29.0 29.2 28.8 23.4 27.8 32.1 28.3 26.2 28.8 32.9 28.9 27.2 32.2 32.3 28.8 44.9 28.9 30.3 25.8 31.2 26.3 25.2 28.7 28.5 30.1 30.1 25.0 77.5 24.7 61.0 29.8 23.1 29.7 17.1 27.0 26.3 27.0 20.0 30.3 30.2 29.3 48.3 29.2 36.3 29.2 28.9 30.9 30.7 31.0 30.7 25.6 90.1 25.8 68.4 27.0 26.7 30.2 43.1 30.2 31.0 27.7 27.2 27.7 47.9 27.7 36.0 28.9 30.0 25.2 25.3 27.6 27.8 28.5 90.3 28.1 68.8 28.6 37.8 29.7 30.8 28.5 27.2 28.6 21.1 30.3 30.1 29.3 50.7 29.9 37.2 27.3 27.7 27.2 22.1 27.7 27.1 28.7 20.2 28.4 31.9 28.5 26.2 31.3 31.5 27.3 52.3 27.2 40.4 30.2 30.0 28.8 42.9 28.4 31.2 25.6 25.4 27.6 27.8 29.1 29.1 30.3 88.0 30.7 67.7 26.7 30.5 26.7 25.0 28.2 29.1 27.7 23.4 27.3 31.2 26.7 25.5 26.2 34.7 26.6 29.3 28.2 28.5 27.4 21.2 28.2 9.4 26.1 26.4 26.1 21.3 26.0 26.2 26.7 53.0 26.9 42.5 23.6 23.0 25.1 43.8 24.6 32.4 Average bodyweight (g) 33.90 30.85 30.70 28.15 30.85 29.45 26.95 30.50 30.80 27.25 33.05 29.90 31.60 32.95 30.90 29.95 32.80 26.55 26.75 28.90 28.05 28.85 32.25 28.85 26.05 28.60 30.10 24.85 29.75 27.00 30.25 29.25 29.05 30.80 30.85 25.70 26.85 30.20 27.45 27.70 29.45 25.25 27.70 28.30 29.15 28.55 30.20 29.60 27.25 28.20 28.45 31.40 27.25 30.10 28.60 25.50 27.70 29.10 30.50 26.70 27.95 27.00 26.40 28.35 27.80 26.10 26.10 26.80 23.30 24.85 Food intake (g) coll. point 1 10.8 12.4 11.6 6.4 16.8 19.9 6.9 20.8 10.5 5.8 5.9 5.7 14.6 6.0 16.5 6.0 6.3 12.0 21.7 12.1 11.9 21.5 7.0 6.1 13.5 5.6 6.9 5.7 11.9 11.7 20.3 5.5 5.7 5.7 5.4 11.8 5.1 10.5 11.4 Page A53 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group# 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 5 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 cage# 1 1 2 2 3 3 4 5 5 5 6 6 6 7 8 8 8 9 9 10 11 12 13 13 14 15 15 15 16 17 18 18 19 19 19 19 20 20 21 21 22 22 22 22 23 24 25 25 26 27 28 29 29 30 30 31 31 31 31 32 33 34 35 36 36 37 38 38 39 39 eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 collection point 2; 3 days feces collection start collection point 2 end collection point 2 bodyweight (g) food on cage (g) bodyweight (g) food off cage (g) 33.9 32.9 30.5 38.9 30.1 22.1 31 30 28.2 39.5 28.3 19.5 30.6 29.2 29.4 35.8 30.4 16.8 27.2 20.5 26.3 10.7 30.6 30 31 30.5 27.3 44.8 27 19.2 33.1 32.2 30 30 32 48.6 31.2 19.8 33.1 24.8 33.6 12.4 31 30.1 29.8 30 32.8 47.2 32.5 14.2 26.6 26.3 26.6 31.8 26.9 13.6 28.8 23.4 29 13.4 28.3 26.2 28.4 15.7 28.9 27.2 29.2 16.2 32.3 32.3 28.9 30.3 29.2 7.1 26.3 25.2 26.1 15.6 28.5 28.8 30.1 29.8 24.7 61.0 25.1 32.7 29.7 17.1 29.4 8.0 27 20.0 27.6 9.4 30.2 29.6 29.2 36.3 28.7 16.5 28.9 29.6 30.7 30.5 30.7 30.7 25.8 68.4 26.2 33.2 26.7 26.9 30.2 31.0 30.1 11.1 27.2 27.2 27.7 36.0 27.7 17.7 30 29.1 25.3 25.5 27.8 28.6 28.1 68.8 28.8 32.3 29.7 30.8 30.1 20.1 28.6 21.1 28.6 10.6 30.1 30.1 29.9 37.2 29.7 15.5 27.2 22.1 27 12.5 28.7 20.2 28.4 10.2 28.5 26.2 28.7 16.6 31.5 31.4 27.2 40.4 27 20.3 30 29.7 28.4 31.2 28 12.3 25.4 25.3 27.8 26.9 29.1 28.8 30.7 67.7 30.9 36.7 26.7 25.0 26.3 16.0 27.7 23.4 29.1 12.7 26.7 25.5 27.8 16.3 26.6 29.3 26.8 20.0 28.5 28.9 28.2 56.1 27.6 38.8 26.1 21.3 26.2 12.1 26.2 26.7 26.9 42.5 27.2 24.9 23 23.6 24.6 32.4 25.7 15.0 Average bodyweight (g) 33.40 30.30 30.50 28.25 29.90 29.90 26.75 30.30 30.75 27.15 32.65 30.00 31.60 33.35 30.55 29.90 32.65 26.45 26.75 28.90 28.35 29.05 32.30 29.05 26.20 28.65 29.95 24.90 29.55 27.30 29.90 28.95 29.25 30.60 30.70 26.00 26.80 30.15 27.20 27.70 29.55 25.40 28.20 28.45 29.90 28.60 30.10 29.80 27.10 28.55 28.60 31.45 27.10 29.85 28.20 25.35 27.35 28.95 30.80 26.50 28.40 27.25 26.70 28.70 27.90 26.15 26.45 27.05 23.30 25.15 Food intake (g) coll. point 2 16.8 20.0 19.0 9.8 25.6 28.8 12.4 33.0 18.2 10.0 10.5 11.0 23.2 9.6 28.3 9.1 10.6 19.8 35.2 19.9 18.3 36.5 10.7 10.5 21.7 9.6 10.0 9.6 20.1 18.9 31.0 9.0 10.7 9.2 9.3 17.3 9.2 17.6 17.4 Page A54 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group# 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 5 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 cage# 1 1 2 2 3 3 4 5 5 5 6 6 6 7 8 8 8 9 9 10 11 12 13 13 14 15 15 15 16 17 18 18 19 19 19 19 20 20 21 21 22 22 22 22 23 24 25 25 26 27 28 29 29 30 30 31 31 31 31 32 33 34 35 36 36 37 38 38 39 39 eartag LR 01 12 00 20 00 02 11 00 02 12 10 21 02 01 10 01 20 00 21 10 01 02 01 11 10 01 20 12 02 10 10 02 00 10 01 02 00 11 00 11 10 01 11 12 12 20 01 11 00 12 01 10 12 10 02 00 01 02 22 21 20 10 02 00 12 00 00 20 10 01 collection point 3; 2 days feces collection start collection point 3 end collection point 3 bodyweight (g) food on cage (g) bodyweight (g) food off cage (g) 32.9 33.2 30.1 78.3 30.2 68.3 30 30.2 28.3 73.4 28.4 63.2 29.2 31.1 30.4 67.0 29.6 56.6 26.3 38.3 26.5 33.4 30 30.8 30.5 30.7 27 94.8 27.1 81.0 32.2 32.8 30 30.0 31.2 97.1 31.8 80.4 33.6 42.0 33.1 36.8 30.1 30.3 30 30.9 32.5 85.6 32.7 67.2 26.3 25.9 26.9 60.9 26.5 52.8 29 46.3 29.1 41.2 28.4 39.9 29.1 34.3 29.2 40.9 29.2 35.5 32.3 32.5 29.2 56.5 29.3 43.8 26.1 46.1 26.4 41.0 28.8 29.1 29.8 30.2 25.1 114.5 25.4 100.9 29.4 34.9 29.6 29.9 27.6 37.9 27.8 33.3 29.6 29.9 28.7 61.5 29.2 50.2 29.6 29.8 30.5 30.5 30.7 31.1 26.2 130.6 26.5 111.9 26.9 28.0 30.1 70.0 30.9 58.0 27.2 27.5 27.7 67.9 27.5 57.3 29.1 29.8 25.5 25.6 28.6 29.2 28.8 138.4 29.1 119.7 30.1 52.3 29.6 47.7 28.6 37.3 28.6 31.9 30.1 30.6 29.7 74.9 30.2 52.3 27 39.9 27.3 35.1 28.4 35.4 28.6 30.3 28.7 44.3 28.7 38.8 31.4 32.1 27 66.8 27.4 55.8 29.7 30.2 28 60.0 28.5 49.9 25.3 25.6 26.9 27.2 28.8 28.6 30.9 142.0 30.6 124.9 26.3 46.7 26.6 41.8 29.1 35.7 28.5 30.7 27.8 42.4 27.8 37.5 26.8 46.1 27.0 40.3 28.9 28.7 27.6 90.6 28.2 81.1 26.2 39.7 27.1 34.2 26.7 26.7 27.2 76.5 27.3 66.5 23.6 23.8 25.7 65.9 25.4 56.1 Average bodyweight (g) 33.05 30.15 30.10 28.35 30.15 30.00 26.40 30.40 30.60 27.05 32.50 30.00 31.50 33.35 30.20 30.45 32.60 26.10 26.70 29.05 28.75 29.20 32.40 29.25 26.25 28.95 30.00 25.25 29.50 27.70 29.75 28.95 29.70 30.50 30.90 26.35 27.45 30.50 27.35 27.60 29.45 25.55 28.90 28.95 29.85 28.60 30.35 29.95 27.15 28.50 28.70 31.75 27.20 29.95 28.25 25.45 27.05 28.70 30.75 26.45 28.80 27.80 26.90 28.80 27.90 26.65 26.70 27.25 23.70 25.55 Food intake (g) coll. point 3 10.0 10.2 10.4 4.9 13.8 16.7 5.2 18.4 8.1 5.1 5.6 5.4 12.7 5.1 13.6 5.0 4.6 11.3 18.7 12.0 10.6 18.7 4.6 5.4 22.6 4.8 5.1 5.5 11.0 10.1 17.1 4.9 5.0 4.9 5.8 9.5 5.5 10.0 9.8 Page A55 of A172 Appendix to TNO-report of 3M-02 Final version Fecal weight Group 1 1 1 1 1 2 2 2 2 2 3 3 3 3 3 3 3 4 4 4 4 5 5 5 5 5 5 6 6 6 6 6 7 7 7 7 7 7 7 cage# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 Feces collection point 1; 2 days collection number of Tube empty Tube full Tube full after freeze drying mice (g) (g) (g) 2 6.6630 8.3248 8.0444 2 6.6490 8.6605 8.3426 2 6.6221 8.6309 8.3229 1 6.6808 7.664 7.4697 3 6.6712 9.4474 8.9658 3 6.6634 9.8825 9.4253 1 6.6794 7.8244 7.6266 3 6.6207 10.2937 9.7316 2 6.6605 8.2584 8.0008 1 6.6630 7.5596 7.4162 1 6.6697 7.5244 7.3942 1 6.6594 7.5056 7.3806 2 6.6379 9.0681 8.6519 1 6.6624 7.5339 7.4049 3 6.6568 9.433 8.9562 1 6.6607 7.5433 7.3604 1 6.6312 7.6683 7.5344 2 6.6765 8.6953 8.3655 4 6.6311 10.3697 9.7743 2 6.6493 8.6134 8.2667 2 6.6603 8.6425 8.2974 4 6.6918 10.4134 9.8868 1 6.6683 7.7517 7.5567 1 6.7028 7.6752 7.5296 2 6.6868 9.2271 8.8008 1 6.6825 7.5842 7.456 1 6.6889 7.8388 7.6669 1 6.6648 7.6251 7.4945 2 6.6617 8.6518 8.3356 2 6.6503 8.6434 8.3383 4 6.6110 9.8507 9.3193 1 6.6541 7.4635 7.3215 1 6.6615 7.3485 7.2406 1 6.6589 7.5622 7.3939 1 6.6820 7.476 7.3376 2 6.6335 8.4094 8.0542 1 6.6727 7.4706 7.3318 2 6.6890 8.3375 8.054 2 6.6706 8.2396 7.9761 freeze dried feces (g) 1.3814 1.6936 1.7008 0.7889 2.2946 2.7619 0.9472 3.1109 1.3403 0.7532 0.7245 0.7212 2.0140 0.7425 2.2994 0.6997 0.9032 1.6890 3.1432 1.6174 1.6371 3.1950 0.8884 0.8268 2.1140 0.7735 0.9780 0.8297 1.6739 1.6880 2.7083 0.6674 0.5791 0.7350 0.6556 1.4207 0.6591 1.3650 1.3055 04-12-2006 Page A56 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group 1 1 1 1 1 2 2 2 2 2 3 3 3 3 3 3 3 4 4 4 4 5 5 5 5 5 5 6 6 6 6 6 7 7 7 7 7 7 7 cage# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 Feces collection point 2; 3 days collection number of Tube empty Tube full Tube full after freeze drying mice (g) (g) (g) 2 6.6728 9.043 8.654 2 6.6313 9.7853 9.3421 2 6.6987 9.5813 9.188 1 6.6499 8.1391 7.9395 3 6.6805 10.6433 10.0911 3 6.6610 11.3687 10.736 1 6.6450 8.426 8.1553 3 6.6479 12.3927 11.5086 2 6.6676 9.2172 8.8482 1 6.6137 8.0501 7.8405 1 6.6564 8.1514 7.945 1 6.6806 8.3368 8.1005 2 6.6554 10.2976 9.7616 1 6.6905 8.1358 7.9299 3 6.6709 11.0804 10.4427 1 6.6500 8.0103 7.7806 1 6.6622 8.2678 8.0649 2 6.6790 9.8571 9.3923 4 6.6452 12.0852 11.2859 2 6.6370 9.933 9.4414 2 6.6464 9.514 9.0495 4 6.6068 12.3762 11.578 1 6.6411 8.2187 7.9954 1 6.6557 8.1263 7.9237 2 6.6822 10.3758 9.8975 1 6.6134 8.0183 7.792 1 6.6799 8.1596 7.914 1 6.6762 8.1471 7.9636 2 6.6989 9.824 9.3799 2 6.6732 9.5294 9.1304 4 6.6621 11.3048 10.6299 1 6.6310 7.9252 7.7404 1 6.6902 8.1846 7.96 1 6.6712 7.9152 7.7309 1 6.6337 7.9533 7.7519 2 6.6633 9.2718 8.8526 1 6.6324 7.8893 7.7077 2 6.6776 9.1802 8.8063 2 6.6697 9.253 8.8469 freeze dried feces (g) 1.9812 2.7108 2.4893 1.2896 3.4106 4.0750 1.5103 4.8607 2.1806 1.2268 1.2886 1.4199 3.1062 1.2394 3.7718 1.1306 1.4027 2.7133 4.6407 2.8044 2.4031 4.9712 1.3543 1.2680 3.2153 1.1786 1.2341 1.2874 2.6810 2.4572 3.9678 1.1094 1.2698 1.0597 1.1182 2.1893 1.0753 2.1287 2.1772 Page A57 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group 1 1 1 1 1 2 2 2 2 2 3 3 3 3 3 3 3 4 4 4 4 5 5 5 5 5 5 6 6 6 6 6 7 7 7 7 7 7 7 cage# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 Feces collection point 3; 2 days collection number of Tube empty Tube full Tube full after freeze drying mice (g) (g) (g) 2 6.6429 8.5821 8.2399 2 6.6725 8.9822 8.6509 2 6.6831 8.9939 8.653 1 6.6645 7.7309 7.5727 3 6.6432 9.9317 9.4021 3 6.6844 10.106 9.6123 1 6.6652 7.8515 7.6749 3 6.6453 10.6479 10.0341 2 6.6397 8.3585 8.1165 1 6.6569 7.7971 7.6376 1 6.6860 7.7179 7.5628 1 6.6590 7.7398 7.58 2 6.6605 9.3189 8.8877 1 6.6334 7.6444 7.4947 3 6.6582 9.6295 9.1847 1 6.6145 7.6351 7.4819 1 6.6692 7.7894 7.631 2 6.6935 9.0966 8.7345 4 6.6306 10.6276 10.077 2 6.6585 9.1542 8.7793 2 6.6901 8.8739 8.5242 4 6.6840 10.9177 10.2953 1 6.6687 7.6866 7.5361 1 6.6635 7.8102 7.6563 2 6.6833 9.43 9.0427 1 6.6333 7.6942 7.5479 1 6.6752 7.7479 7.5966 1 6.6351 7.7734 7.6365 2 6.6633 9.0885 8.5695 2 6.6611 8.8705 8.5556 4 6.6657 10.3688 9.9106 1 6.6786 7.7434 7.5484 1 6.6860 7.6314 7.485 1 6.6887 7.5625 7.393 1 6.6783 7.9028 7.6867 2 6.6551 8.561 8.2858 1 6.6481 7.617 7.4543 2 6.6522 8.6462 8.3568 2 6.6641 8.7084 8.3702 freeze dried feces (g) 1.5970 1.9784 1.9699 0.9082 2.7589 2.9279 1.0097 3.3888 1.4768 0.9807 0.8768 0.9210 2.2272 0.8613 2.5265 0.8674 0.9618 2.0410 3.4464 2.1208 1.8341 3.6113 0.8674 0.9928 2.3594 0.9146 0.9214 1.0014 1.9062 1.8945 3.2449 0.8698 0.7990 0.7043 1.0084 1.6307 0.8062 1.7046 1.7061 Page A58 of A172 Appendix to TNO-report of 3M-02 Final version Feces weight, bodyweight and food intake GC samples GC-1 code 1:1-1 1:1-2 1:1-3 1:2-4 1:2-5 1:2-6 1:3-7 1:3-8 1:3-9 1:4-10 1:4-11 1:4-12 1:5-13 1:5-14 1:5-15 1:6-16 1:6-17 1:6-18 1:7-19 1:7-20 1:7-21 Feces collection 3M-02 week 6; collection point 1, 2 days collection Group cage# feces collected Bodyweight Food intake (g) (g) (g) 1 1,2 3.075 123.6 23.2 1 3,4 2.4897 87.25 18 1 5 2.2946 88.55 16.8 2 6 2.7619 94.55 19.9 2 7,8 4.0581 126.6 27.7 2 9,10 2.0935 82.2 16.3 3 11,12,13 3.4597 118 26.2 3 15 2.2994 83.55 16.5 3 14,16,17 2.3454 82.8 18.3 4 18 1.689 59.5 12 4 19 3.1432 116.4 21.7 4 20,21 3.2545 112.2 24 5 22 3.195 110.7 21.5 5 24,25 2.9408 88.35 19.6 5 23,26,27 2.6399 84.6 19.5 6 28,29 2.5036 87.1 17.6 6 31 2.7083 112.8 20.3 6 30,32 2.3554 85.4 17.2 7 33,34,35 1.9697 81.35 16.8 7 36,37 2.0798 82.25 16.9 7 38,39 2.6705 101.05 21.9 GC-2 code 2:1-1 2:1-2 2:1-3 2:2-4 2:2-5 2:2-6 2:3-7 2:3-8 2:3-9 2:4-10 2:4-11 2:4-12 2:5-13 2:5-14 2:5-15 2:6-16 2:6-17 2:6-18 2:7-19 2:7-20 2:7-21 Feces collection 3M-02 week 6; collection point 2, 3 days collection Group cage# feces collected Bodyweight Food intake (g) (g) (g) 1 1,2 4.692 122.45 36.8 1 3,4 3.7789 86.55 28.8 1 5 3.4106 88.2 25.6 2 6 4.075 94.25 28.8 2 7,8 6.371 126.45 45.4 2 9,10 3.4074 82.1 28.2 3 11,12,13 5.8147 118.75 44.7 3 15 3.7718 83.5 28.3 3 14,16,17 3.7727 83.05 29.3 4 18 2.7133 58.85 19.8 4 19 4.6407 116.55 35.2 4 20,21 5.2075 111.85 38.2 5 22 4.9712 111.6 36.5 5 24,25 4.4833 88.5 32.2 5 23,26,27 3.767 85.55 30.3 6 28,29 3.9684 87.15 29.7 6 31 3.9678 112.45 31 6 30,32 3.5666 84.55 27.9 7 33,34,35 3.4477 82.35 29.2 7 36,37 3.2646 82.75 26.5 7 38,39 4.3059 101.95 35 GC-3 code 3:1-1 3:1-2 3:1-3 3:2-4 3:2-5 3:2-6 3:3-7 3:3-8 3:3-9 3:4-10 3:4-11 3:4-12 3:5-13 3:5-14 3:5-15 3:6-16 3:6-17 3:6-18 3:7-19 3:7-20 3:7-21 Feces collection 3M-02 week 6; collection point 3, 2 days collection Group cage# feces collected Bodyweight Food intake (g) (g) (g) 1 1,2 3.5754 121.65 20.2 1 3,4 2.8781 86.55 15.3 1 5 2.7589 88.05 13.8 2 6 2.9279 94 16.7 2 7,8 4.3985 126.6 23.6 2 9,10 2.4575 81.85 13.2 3 11,12,13 4.025 119.6 23.7 3 15 2.5265 84.2 13.6 3 14,16,17 2.6905 83.45 14.7 4 18 2.041 58.7 11.3 4 19 3.4464 117.45 18.7 4 20,21 3.9549 112.9 22.6 5 22 3.6113 112.85 18.7 5 24,25 3.3522 88.9 28 5 23,26,27 2.7034 85.5 14.5 6 28,29 2.9076 87.65 16.5 6 31 3.2449 111.95 17.1 6 30,32 2.7643 84.65 15 7 33,34,35 2.5117 83.5 15.7 7 36,37 2.4369 83.35 15 7 38,39 3.4107 103.2 19.8 04-12-2006 Page A59 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 E2. Fecal bile acid measurements 1:1-1 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso 5.8 area 770 2880 837 1412 3635 3854 411 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 1062.6 2880 979 1412 3925.8 5704 411 % g bile acid ng bile acid/ nmol bile acid/ mg feces mg feces 6.5 0.447 77.1 0.189 17.6 1.211 208.9 0.532 6.0 0.412 71.0 0.174 8.6 0.594 102.4 0.272 24.0 1.651 284.7 0.697 34.8 2.399 413.6 1.012 2.5 0.173 29.8 0.076 total 13799 16375 100 7 1187 3.0 nor-hyodeoxy 23775 1 23775 23775 1:1-2 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso 6 area 481 1551 1143 2203 2222 5717 261 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 663.78 1551 1337 2203 2399.76 8461 261 % g bile acid ng bile acid/ nmol bile acid/ mg feces mg feces 3.9 0.276 46.0 0.113 9.2 0.645 107.4 0.274 7.9 0.556 92.6 0.227 13.1 0.916 152.6 0.405 14.2 0.997 166.2 0.407 50.1 3.517 586.1 1.435 1.5 0.108 18.1 0.046 total 13578 16877 100 7 1169 2.9 nor-hyodeoxy 24059 1 24059 24059 1:1-3 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso 5.5 area 304 1047 535 1334 1317 3031 192 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 419.52 1047 626 1334 1422.36 4486 192 % g bile acid ng bile acid/ nmol bile acid/ mg feces mg feces 4.4 0.269 48.9 0.120 11.0 0.672 122.1 0.311 6.6 0.401 73.0 0.179 14.0 0.856 155.6 0.413 14.9 0.912 165.9 0.406 47.1 2.877 523.1 1.280 2.0 0.123 22.4 0.057 total 7760 9527 100 6 1111 2.8 nor-hyodeoxy 15591 1 15591 15591 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A60 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 2:1-1 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:1-2 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:1-3 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 7.2 area 314 688 213 335 803 507 2860 4612 7.2 area 1014 2554 2363 2730 4506 9183 213 22563 29667 5.1 area 1430 3201 3202 808 5434 7559 255 21889 43696 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 433.32 688 249 335 867.24 750 0 3323 1 4612 % 13.0 20.7 7.5 10.1 26.1 22.6 0.0 100 4612 g bile acid ng bile acid/ nmol bile acid/ mg feces mg feces 0.940 130.5 0.319 1.492 207.2 0.528 0.540 75.0 0.184 0.726 100.9 0.268 1.880 261.2 0.639 1.627 226.0 0.553 0.000 0.0 0.000 7 1001 2.5 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1399.32 5.0 0.472 65.5 0.160 1 2554 9.1 0.861 119.6 0.305 1 2765 9.8 0.932 129.4 0.317 1 2730 9.7 0.920 127.8 0.339 1.08 4866.48 17.3 1.640 227.8 0.558 1 13591 48.3 4.581 636.3 1.557 1 213 0.8 0.072 10.0 0.025 28118 100 9 1316 3.3 1 29667 29667 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1973.4 7.3 0.452 88.6 0.217 1 3201 11.8 0.733 143.6 0.366 1 3746 13.9 0.857 168.1 0.411 1 808 3.0 0.185 36.3 0.096 1.08 5868.72 21.7 1.343 263.3 0.645 1 11187 41.4 2.560 502.0 1.229 1 255 0.9 0.058 11.4 0.029 27040 100 6 1213 3.0 1 43696 43696 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A61 of A172 Appendix to TNO-report of 3M-02 Final version 3:1-1 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:1-2 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:1-3 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 7 area 755 6854 1031 1211 6883 6151 349 23234 32626 6.6 area 475 1558 748 1489 1894 3897 198 10259 20195 6.1 area 750 1906 1166 974 3492 4213 396 12897 24103 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1041.9 3.8 0.319 45.6 0.112 1 6854 25.2 2.101 300.1 0.764 1 1206 4.4 0.370 52.8 0.129 1 1211 4.5 0.371 53.0 0.141 1.08 7433.64 27.3 2.278 325.5 0.797 1 9103 33.5 2.790 398.6 0.976 1 349 1.3 0.107 15.3 0.039 27199 100 8 1191 3.0 1 32626 32626 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 655.5 5.2 0.325 49.2 0.120 1 1558 12.4 0.771 116.9 0.298 1 875 7.0 0.433 65.7 0.161 1 1489 11.8 0.737 111.7 0.297 1.08 2045.52 16.2 1.013 153.5 0.376 1 5768 45.8 2.856 432.7 1.059 1 198 1.6 0.098 14.9 0.038 12589 100 6 944 2.3 1 20195 20195 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1035 6.6 0.429 70.4 0.172 1 1906 12.2 0.791 129.6 0.330 1 1364 8.7 0.566 92.8 0.227 1 974 6.2 0.404 66.2 0.176 1.08 3771.36 24.0 1.565 256.5 0.628 1 6235 39.8 2.587 424.1 1.038 1 396 2.5 0.164 26.9 0.069 15682 100 7 1067 2.6 1 24103 24103 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A62 of A172 Appendix to TNO-report of 3M-02 Final version 1:2-4 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:2-5 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:2-6 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 6.3 area 133 741 488 509 758 1756 133 4518 8688 6.4 area 385 1785 1437 1980 5071 4256 151 15065 25522 5.3 area 696 3055 1508 3344 2771 7511 363 19248 29599 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 183.54 741 571 509 818.64 2599 133 5555 1 8688 % 3.3 13.3 10.3 9.2 14.7 46.8 2.4 100 8688 g bile acid ng bile acid/ nmol bile acid/ mg feces mg feces 0.211 33.5 0.082 0.853 135.4 0.345 0.657 104.3 0.255 0.586 93.0 0.247 0.942 149.6 0.366 2.991 474.8 1.162 0.153 24.3 0.062 6 1015 2.5 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 531.3 3.0 0.208 32.5 0.080 1 1785 10.0 0.699 109.3 0.278 1 1681 9.4 0.659 102.9 0.252 1 1980 11.1 0.776 121.2 0.322 1.08 5476.68 30.6 2.146 335.3 0.821 1 6299 35.2 2.468 385.6 0.944 1 151 0.8 0.059 9.2 0.024 17904 100 7 1096 2.7 1 25522 25522 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 960.48 4.1 0.324 61.2 0.150 1 3055 12.9 1.032 194.7 0.496 1 1764 7.5 0.596 112.5 0.275 1 3344 14.2 1.130 213.2 0.566 1.08 2992.68 12.7 1.011 190.8 0.467 1 11116 47.1 3.756 708.6 1.734 1 363 1.5 0.123 23.1 0.059 23596 100 8 1504 3.7 1 29599 29599 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A63 of A172 Appendix to TNO-report of 3M-02 Final version 2:2-4 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:2-5 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:2-6 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 7.7 area 564 1799 924 1298 1925 3892 10402 15018 6.6 area 521 1246 771 1507 3321 3749 322 11437 18400 6.4 area 1341 4170 1935 3343 3407 9002 474 23672 29674 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 778.32 6.1 0.518 67.3 0.165 1 1799 14.1 1.198 155.6 0.396 1 1081 8.4 0.720 93.5 0.229 1 1298 10.1 0.864 112.2 0.298 1.08 2079 16.2 1.384 179.8 0.440 1 5760 45.0 3.836 498.1 1.219 1 0 0.0 0.000 0.0 0.000 12796 100 9 1107 2.7 1 15018 15018 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 718.98 5.2 0.391 59.2 0.145 1 1246 9.0 0.677 102.6 0.261 1 902 6.5 0.490 74.3 0.182 1 1507 10.9 0.819 124.1 0.330 1.08 3586.68 25.9 1.949 295.3 0.723 1 5549 40.1 3.016 456.9 1.118 1 322 2.3 0.175 26.5 0.068 13831 100 8 1139 2.8 1 18400 18400 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1850.58 6.4 0.624 97.4 0.238 1 4170 14.3 1.405 219.6 0.559 1 2264 7.8 0.763 119.2 0.292 1 3343 11.5 1.127 176.0 0.467 1.08 3679.56 12.6 1.240 193.7 0.474 1 13323 45.8 4.490 701.5 1.717 1 474 1.6 0.160 25.0 0.064 29104 100 10 1532 3.8 1 29674 29674 04-12-2006 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A64 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 3:2-4 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:2-5 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:2-6 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 6.4 area 469 1577 949 839 2511 4030 280 10655 16647 6.5 area 438 1197 858 906 3408 2988 364 10159 18855 5.5 area 871 2878 1791 1999 2933 6248 527 17247 26219 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 647.22 4.9 0.389 60.7 0.149 1 1577 12.0 0.947 148.0 0.377 1 1110 8.5 0.667 104.2 0.255 1 839 6.4 0.504 78.7 0.209 1.08 2711.88 20.7 1.629 254.5 0.623 1 5964 45.4 3.583 559.8 1.370 1 280 2.1 0.168 26.3 0.067 13130 100 8 1232 3.0 1 16647 16647 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 604.44 5.0 0.321 49.3 0.121 1 1197 9.8 0.635 97.7 0.249 1 1004 8.2 0.532 81.9 0.200 1 906 7.4 0.481 73.9 0.196 1.08 3680.64 30.2 1.952 300.3 0.735 1 4422 36.3 2.345 360.8 0.883 1 364 3.0 0.193 29.7 0.076 12178 100 6 994 2.5 1 18855 18855 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1201.98 5.7 0.458 83.4 0.204 1 2878 13.6 1.098 199.6 0.508 1 2095 9.9 0.799 145.3 0.356 1 1999 9.5 0.762 138.6 0.368 1.08 3167.64 15.0 1.208 219.7 0.538 1 9247 43.8 3.527 641.2 1.569 1 527 2.5 0.201 36.5 0.093 21116 100 8 1464 3.6 1 26219 26219 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A65 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 1:3-7 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:3-8 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:3-9 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5.8 area 463 2160 560 2503 3081 5475 269 14511 22598 5.4 area 258 1480 573 1667 1531 3397 212 9118 15759 6.4 area 483 1604 840 2300 2706 3466 11399 16088 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 638.94 3.6 0.283 48.7 0.119 1 2160 12.2 0.956 164.8 0.420 1 655 3.7 0.290 50.0 0.122 1 2503 14.2 1.108 191.0 0.507 1.08 3327.48 18.8 1.472 253.9 0.621 1 8103 45.9 3.586 618.2 1.513 1 269 1.5 0.119 20.5 0.052 17657 100 8 1347 3.4 1 22598 22598 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 356.04 3.2 0.226 41.8 0.102 1 1480 13.4 0.939 173.9 0.443 1 670 6.1 0.425 78.8 0.193 1 1667 15.1 1.058 195.9 0.520 1.08 1653.48 14.9 1.049 194.3 0.476 1 5028 45.4 3.190 590.8 1.446 1 212 1.9 0.135 24.9 0.063 11066 100 7 1300 3.2 1 15759 15759 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 666.54 4.9 0.414 64.7 0.158 1 1604 11.8 0.997 155.8 0.397 1 983 7.2 0.611 95.5 0.234 1 2300 16.9 1.430 223.4 0.593 1.08 2922.48 21.5 1.817 283.8 0.695 1 5130 37.7 3.189 498.2 1.219 1 0 0.0 0.000 0.0 0.000 13606 100 8 1321 3.3 1 16088 16088 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A66 of A172 Appendix to TNO-report of 3M-02 Final version 2:3-7 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:3-8 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:3-9 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5 area 213 1811 1070 2080 2392 4005 367 11938 21102 7.3 area 222 790 602 1050 1101 2234 135 6134 10229 5.3 area 887 2496 1947 482 4734 4874 15420 36744 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 293.94 2.1 0.139 27.9 0.068 1 1811 12.7 0.858 171.6 0.437 1 1252 8.7 0.593 118.7 0.290 1 2080 14.5 0.986 197.1 0.523 1.08 2583.36 18.0 1.224 244.8 0.599 1 5927 41.4 2.809 561.8 1.375 1 367 2.6 0.174 34.8 0.089 14315 100 7 1357 3.4 1 21102 21102 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 306.36 4.1 0.300 41.0 0.100 1 790 10.6 0.772 105.8 0.269 1 704 9.4 0.689 94.3 0.231 1 1050 14.0 1.026 140.6 0.373 1.08 1189.08 15.9 1.162 159.2 0.390 1 3306 44.2 3.232 442.8 1.084 1 135 1.8 0.132 18.1 0.046 7481 100 7 1002 2.5 1 10229 10229 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1224.06 6.5 0.333 62.9 0.154 1 2496 13.3 0.679 128.2 0.326 1 2278 12.1 0.620 117.0 0.286 1 482 2.6 0.131 24.8 0.066 1.08 5112.72 27.2 1.391 262.5 0.643 1 7214 38.4 1.963 370.4 0.907 1 0 0.0 0.000 0.0 0.000 18806 100 5 966 2.4 1 36744 36744 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A67 of A172 Appendix to TNO-report of 3M-02 Final version 3:3-7 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:3-8 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:3-9 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 8.2 area 727 3031 1265 1974 4421 5331 401 17150 26440 6.4 area 541 1842 998 1904 2601 4796 270 12952 28268 7 area 612 2141 1127 2027 3995 4490 224 14616 22149 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1003.26 4.9 0.379 46.3 0.113 1 3031 14.7 1.146 139.8 0.356 1 1480 7.2 0.560 68.3 0.167 1 1974 9.6 0.747 91.0 0.242 1.08 4774.68 23.2 1.806 220.2 0.539 1 7890 38.4 2.984 363.9 0.891 1 401 2.0 0.152 18.5 0.047 20554 100 8 948 2.4 1 26440 26440 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 746.58 4.7 0.264 41.3 0.101 1 1842 11.6 0.652 101.8 0.259 1 1168 7.4 0.413 64.5 0.158 1 1904 12.0 0.674 105.2 0.279 1.08 2809.08 17.7 0.994 155.3 0.380 1 7098 44.8 2.511 392.3 0.960 1 270 1.7 0.096 14.9 0.038 15837 100 6 875 2.2 1 28268 28268 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 844.56 4.8 0.381 54.5 0.133 1 2141 12.2 0.967 138.1 0.352 1 1319 7.5 0.595 85.0 0.208 1 2027 11.6 0.915 130.7 0.347 1.08 4314.6 24.6 1.948 278.3 0.681 1 6645 37.9 3.000 428.6 1.049 1 224 1.3 0.101 14.4 0.037 17515 100 8 1130 2.8 1 22149 22149 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A68 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 1:4-10 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:4-11 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:4-12 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5.1 area 103 354 286 394 349 1185 235 2906 12494 4.8 area 276 396 184 214 555 896 2521 8389 6 area 320 852 475 954 954 2059 5614 11311 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 142.14 4.0 0.114 22.3 0.055 1 354 9.9 0.283 55.6 0.142 1 335 9.3 0.268 52.5 0.129 1 394 11.0 0.315 61.8 0.164 1.08 376.92 10.5 0.302 59.2 0.145 1 1754 48.8 1.404 275.2 0.674 1 235 6.5 0.188 36.9 0.094 3590 100 3 563 1.4 1 12494 12494 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 380.88 396 215 214 599.4 1326 0 3132 1 8389 % 12.2 12.6 6.9 6.8 19.1 42.3 0.0 100 8389 g bile acid 0.454 0.472 0.257 0.255 0.715 1.581 0.000 ng bile acid/ mg feces 94.6 98.3 53.5 53.1 148.9 329.3 0.0 nmol bile acid/ mg feces 0.231 0.250 0.131 0.141 0.364 0.806 0.000 4 778 1.9 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 441.6 6.4 0.390 65.1 0.159 1 852 12.4 0.753 125.5 0.320 1 556 8.1 0.491 81.9 0.200 1 954 13.9 0.843 140.6 0.373 1.08 1030.32 15.0 0.911 151.8 0.372 1 3047 44.3 2.694 449.0 1.099 1 0 0.0 0.000 0.0 0.000 6881 100 6 1014 2.5 1 11311 11311 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A69 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 2:4-10 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:4-11 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:4-12 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 6.2 area 361 1051 1247 1420 979 2197 7255 19330 6.6 area 842 1126 866 1217 2527 3746 481 10805 22800 7.6 area 543 902 1180 1019 1622 2624 206 8096 13502 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 498.18 5.7 0.258 41.6 0.102 1 1051 12.0 0.544 87.7 0.223 1 1459 16.7 0.755 121.7 0.298 1 1420 16.3 0.735 118.5 0.315 1.08 1057.32 12.1 0.547 88.2 0.216 1 3252 37.2 1.682 271.3 0.664 1 0 0.0 0.000 0.0 0.000 8737 100 5 729 1.8 1 19330 19330 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1161.96 8.8 0.510 77.2 0.189 1 1126 8.5 0.494 74.8 0.191 1 1013 7.6 0.444 67.3 0.165 1 1217 9.2 0.534 80.9 0.215 1.08 2729.16 20.6 1.197 181.4 0.444 1 5544 41.8 2.432 368.4 0.902 1 481 3.6 0.211 32.0 0.081 13272 100 6 882 2.2 1 22800 22800 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 749.34 7.6 0.555 73.0 0.179 1 902 9.1 0.668 87.9 0.224 1 1381 14.0 1.023 134.5 0.329 1 1019 10.3 0.755 99.3 0.264 1.08 1751.76 17.7 1.297 170.7 0.418 1 3884 39.3 2.876 378.5 0.926 1 206 2.1 0.153 20.1 0.051 9892 100 7 964 2.4 1 13502 13502 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A70 of A172 Appendix to TNO-report of 3M-02 Final version 3:4-10 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:4-11 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:4-12 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 8.1 area 374 928 737 749 1189 2355 244 6576 16790 8.3 area 474 1184 335 668 2030 2413 257 7361 13770 6.6 area 436 831 1182 693 1774 2161 172 7249 15281 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 516.12 6.4 0.307 38.0 0.093 1 928 11.5 0.553 68.2 0.174 1 862 10.7 0.514 63.4 0.155 1 749 9.3 0.446 55.1 0.146 1.08 1284.12 15.9 0.765 94.4 0.231 1 3485 43.2 2.076 256.3 0.627 1 244 3.0 0.145 17.9 0.046 8069 100 5 593 1.5 1 16790 16790 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 654.12 7.3 0.475 57.2 0.140 1 1184 13.3 0.860 103.6 0.264 1 392 4.4 0.285 34.3 0.084 1 668 7.5 0.485 58.4 0.155 1.08 2192.4 24.6 1.592 191.8 0.469 1 3571 40.0 2.593 312.5 0.765 1 257 2.9 0.187 22.5 0.057 8919 100 6 780 1.9 1 13770 13770 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 601.68 6.8 0.394 59.7 0.146 1 831 9.4 0.544 82.4 0.210 1 1383 15.7 0.905 137.1 0.336 1 693 7.9 0.454 68.7 0.182 1.08 1915.92 21.8 1.254 190.0 0.465 1 3198 36.4 2.093 317.1 0.776 1 172 2.0 0.113 17.1 0.043 8795 100 6 872 2.2 1 15281 15281 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A71 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 1:5-13 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:5-14 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:5-15 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5.1 area 279 1398 233 1493 1698 1962 138 7201 19809 5.9 area 273 1027 1007 1318 1369 1924 193 7111 18666 6.6 area 201 976 457 1197 374 2027 5232 11586 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 385.02 4.6 0.194 38.1 0.093 1 1398 16.6 0.706 138.4 0.352 1 273 3.2 0.138 27.0 0.066 1 1493 17.7 0.754 147.8 0.392 1.08 1833.84 21.8 0.926 181.5 0.444 1 2904 34.5 1.466 287.4 0.703 1 138 1.6 0.070 13.7 0.035 8424 100 4 834 2.1 1 19809 19809 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 376.74 4.5 0.202 34.2 0.084 1 1027 12.2 0.550 93.3 0.238 1 1178 14.0 0.631 107.0 0.262 1 1318 15.7 0.706 119.7 0.318 1.08 1478.52 17.6 0.792 134.3 0.329 1 2848 33.8 1.526 258.6 0.633 1 193 2.3 0.103 17.5 0.045 8419 100 5 764 1.9 1 18666 18666 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 277.38 4.3 0.239 36.3 0.089 1 976 15.3 0.842 127.6 0.325 1 535 8.4 0.461 69.9 0.171 1 1197 18.7 1.033 156.5 0.416 1.08 403.92 6.3 0.349 52.8 0.129 1 3000 47.0 2.589 392.3 0.960 1 0 0.0 0.000 0.0 0.000 6389 100 6 836 2.1 1 11586 11586 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A72 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 2:5-13 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:5-14 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:5-15 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 6.9 area 170 421 188 1014 816 778 196 3583 8118 7.1 area 488 1599 1339 1652 2345 2835 357 10615 19702 5.4 area 1147 3831 1371 3141 2207 7603 531 19831 34357 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 234.6 421 220 1014 881.28 1151 196 4118 1 8118 % 5.7 10.2 5.3 24.6 21.4 28.0 4.8 100 8118 g bile acid 0.289 0.519 0.271 1.249 1.086 1.418 0.241 ng bile acid/ mg feces 41.9 75.2 39.3 181.0 157.3 205.6 35.0 nmol bile acid/ mg feces 0.103 0.191 0.096 0.481 0.385 0.503 0.089 5 735 1.8 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 673.44 5.4 0.342 48.1 0.118 1 1599 12.7 0.812 114.3 0.291 1 1567 12.5 0.795 112.0 0.274 1 1652 13.1 0.838 118.1 0.314 1.08 2532.6 20.1 1.285 181.0 0.443 1 4196 33.4 2.130 299.9 0.734 1 357 2.8 0.181 25.5 0.065 12576 100 6 899 2.2 1 19702 19702 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1582.86 6.5 0.461 85.3 0.209 1 3831 15.7 1.115 206.5 0.526 1 1604 6.6 0.467 86.5 0.212 1 3141 12.9 0.914 169.3 0.450 1.08 2383.56 9.8 0.694 128.5 0.314 1 11252 46.3 3.275 606.5 1.484 1 531 2.2 0.155 28.6 0.073 24326 100 7 1311 3.3 1 34357 34357 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A73 of A172 Appendix to TNO-report of 3M-02 Final version 3:5-13 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:5-14 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:5-15 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 7.9 area 608 1723 925 1896 2481 3000 400 11033 29729 8.1 area 391 1007 1069 799 1999 2126 548 7939 20411 7.3 area 908 3513 1190 2079 2885 6815 481 17871 34491 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 839.04 6.4 0.282 35.7 0.087 1 1723 13.2 0.580 73.4 0.187 1 1082 8.3 0.364 46.1 0.113 1 1896 14.5 0.638 80.7 0.214 1.08 2679.48 20.5 0.901 114.1 0.279 1 4440 34.0 1.493 189.0 0.463 1 400 3.1 0.135 17.0 0.043 13060 100 4 556 1.4 1 29729 29729 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 539.58 5.7 0.264 32.6 0.080 1 1007 10.7 0.493 60.9 0.155 1 1251 13.2 0.613 75.7 0.185 1 799 8.5 0.391 48.3 0.128 1.08 2158.92 22.8 1.058 130.6 0.320 1 3146 33.3 1.542 190.3 0.466 1 548 5.8 0.268 33.1 0.084 9450 100 5 572 1.4 1 20411 20411 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1253.04 5.7 0.363 49.8 0.122 1 3513 16.0 1.019 139.5 0.355 1 1392 6.4 0.404 55.3 0.135 1 2079 9.5 0.603 82.6 0.219 1.08 3115.8 14.2 0.903 123.7 0.303 1 10086 46.0 2.924 400.6 0.980 1 481 2.2 0.139 19.1 0.049 21920 100 6 871 2.2 1 34491 34491 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A74 of A172 Appendix to TNO-report of 3M-02 Final version 1:6-16 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:6-17 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:6-18 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5.8 area 222 654 724 565 752 1281 123 4321 16312 6.2 area 199 543 411 518 383 1077 111 3242 10983 6.3 area 117 650 962 598 677 1575 174 4753 13455 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 306.36 5.9 0.188 32.4 0.079 1 654 12.6 0.401 69.1 0.176 1 847 16.3 0.519 89.5 0.219 1 565 10.9 0.346 59.7 0.159 1.08 812.16 15.6 0.498 85.8 0.210 1 1896 36.4 1.162 200.4 0.490 1 123 2.4 0.075 13.0 0.033 5203 100 3 550 1.4 1 16312 16312 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 274.62 7.0 0.250 40.3 0.099 1 543 13.8 0.494 79.7 0.203 1 481 12.2 0.438 70.6 0.173 1 518 13.2 0.472 76.1 0.202 1.08 413.64 10.5 0.377 60.7 0.149 1 1594 40.5 1.451 234.1 0.573 1 111 2.8 0.101 16.3 0.042 3935 100 4 578 1.4 1 10983 10983 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 161.46 2.8 0.120 19.0 0.047 1 650 11.3 0.483 76.7 0.195 1 1126 19.5 0.837 132.8 0.325 1 598 10.4 0.444 70.5 0.187 1.08 731.16 12.7 0.543 86.3 0.211 1 2331 40.4 1.732 275.0 0.673 1 174 3.0 0.129 20.5 0.052 5771 100 4 681 1.7 1 13455 13455 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A75 of A172 Appendix to TNO-report of 3M-02 Final version 2:6-16 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:6-17 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:6-18 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5.6 area 215 821 747 598 840 1244 226 4691 17522 5.5 area 196 966 220 486 177 733 2778 11207 7.4 area 553 1729 1319 1154 1500 3330 318 9903 20902 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 296.7 5.3 0.169 30.2 0.074 1 821 14.8 0.469 83.7 0.213 1 874 15.7 0.499 89.1 0.218 1 598 10.7 0.341 60.9 0.162 1.08 907.2 16.3 0.518 92.5 0.226 1 1841 33.1 1.051 187.6 0.459 1 226 4.1 0.129 23.0 0.059 5564 100 3 567 1.4 1 17522 17522 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 270.48 8.3 0.241 43.9 0.107 1 966 29.7 0.862 156.7 0.399 1 257 7.9 0.230 41.8 0.102 1 486 14.9 0.434 78.8 0.209 1.08 191.16 5.9 0.171 31.0 0.076 1 1085 33.3 0.968 176.0 0.431 1 0 0.0 0.000 0.0 0.000 3256 100 3 528 1.3 1 11207 11207 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 763.14 6.3 0.365 49.3 0.121 1 1729 14.3 0.827 111.8 0.285 1 1543 12.8 0.738 99.8 0.244 1 1154 9.6 0.552 74.6 0.198 1.08 1620 13.4 0.775 104.7 0.256 1 4928 40.9 2.358 318.6 0.780 1 318 2.6 0.152 20.6 0.052 12056 100 6 779 1.9 1 20902 20902 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A76 of A172 Appendix to TNO-report of 3M-02 Final version 3:6-16 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:6-17 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 3:6-18 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 7.4 area 621 1219 1628 685 1439 1866 253 7711 23222 8.1 area 829 1831 1171 653 1120 4227 295 10126 26821 7.7 area 441 1194 995 999 934 1799 6362 24030 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 856.98 9.3 0.369 49.9 0.122 1 1219 13.2 0.525 70.9 0.181 1 1905 20.6 0.820 110.8 0.271 1 685 7.4 0.295 39.9 0.106 1.08 1554.12 16.8 0.669 90.4 0.221 1 2762 29.9 1.189 160.7 0.393 1 253 2.7 0.109 14.7 0.038 9235 100 4 537 1.3 1 23222 23222 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1144.02 9.0 0.427 52.7 0.129 1 1831 14.4 0.683 84.3 0.215 1 1370 10.7 0.511 63.1 0.154 1 653 5.1 0.243 30.1 0.080 1.08 1209.6 9.5 0.451 55.7 0.136 1 6256 49.0 2.332 288.0 0.705 1 295 2.3 0.110 13.6 0.035 12759 100 5 587 1.5 1 26821 26821 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 608.58 8.0 0.253 32.9 0.080 1 1194 15.6 0.497 64.5 0.164 1 1164 15.2 0.484 62.9 0.154 1 999 13.1 0.416 54.0 0.143 1.08 1008.72 13.2 0.420 54.5 0.133 1 2663 34.9 1.108 143.9 0.352 1 0 0.0 0.000 0.0 0.000 7637 100 3 413 1.0 1 24030 24030 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A77 of A172 Appendix to TNO-report of 3M-02 Final version 1:7-19 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:7-20 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 1:7-21 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5.2 area 493 1997 1424 376 1518 1175 6983 12241 5.8 area 602 895 817 302 659 977 267 4519 11457 5.4 area 512 1419 1545 649 893 912 204 6134 13452 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 680.34 8.4 0.556 106.9 0.262 1 1997 24.7 1.631 313.7 0.799 1 1666 20.6 1.361 261.7 0.641 1 376 4.6 0.307 59.1 0.157 1.08 1639.44 20.2 1.339 257.6 0.630 1 1739 21.5 1.421 273.2 0.669 1 0 0.0 0.000 0.0 0.000 8098 100 7 1272 3.2 1 12241 12241 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 830.76 15.4 0.725 125.0 0.306 1 895 16.5 0.781 134.7 0.343 1 956 17.7 0.834 143.8 0.352 1 302 5.6 0.264 45.4 0.121 1.08 711.72 13.2 0.621 107.1 0.262 1 1446 26.7 1.262 217.6 0.533 1 267 4.9 0.233 40.2 0.102 5408 100 5 814 2.0 1 11457 11457 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 706.56 10.0 0.525 97.3 0.238 1 1419 20.0 1.055 195.3 0.498 1 1808 25.5 1.344 248.8 0.609 1 649 9.1 0.482 89.3 0.237 1.08 964.44 13.6 0.717 132.8 0.325 1 1350 19.0 1.003 185.8 0.455 1 204 2.9 0.152 28.1 0.072 7100 100 5 977 2.4 1 13452 13452 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A78 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 2:7-19 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:7-20 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 2:7-21 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 5.5 area 216 631 176 110 543 386 2062 5496 5.9 area 872 3015 1451 382 2004 3004 653 11381 27158 5.3 area 665 3474 1201 397 1600 2024 155 9516 15683 mg response factor 1.38 1 1 1 1.08 1 1 area corrected for response factor 298.08 631 206 110 586.44 571 0 2403 1 5496 % 12.4 26.3 8.6 4.6 24.4 23.8 0.0 100 5496 g bile acid 0.542 1.148 0.375 0.200 1.067 1.039 0.000 ng bile acid/ mg feces 98.6 208.7 68.1 36.4 194.0 189.0 0.0 nmol bile acid/ mg feces 0.241 0.532 0.167 0.097 0.475 0.463 0.000 4 795 2.0 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1203.36 8.9 0.443 75.1 0.184 1 3015 22.2 1.110 188.2 0.479 1 1698 12.5 0.625 106.0 0.259 1 382 2.8 0.141 23.8 0.063 1.08 2164.32 16.0 0.797 135.1 0.331 1 4446 32.8 1.637 277.5 0.679 1 653 4.8 0.240 40.8 0.104 13561 100 5 846 2.1 1 27158 27158 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 917.7 8.3 0.585 110.4 0.270 1 3474 31.4 2.215 418.0 1.065 1 1405 12.7 0.896 169.1 0.414 1 397 3.6 0.253 47.8 0.127 1.08 1728 15.6 1.102 207.9 0.509 1 2996 27.1 1.910 360.4 0.882 1 155 1.4 0.099 18.6 0.047 11072 100 7 1332 3.3 1 15683 15683 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 Page A79 of A172 Appendix to TNO-report of 3M-02 Final version 3:7-19 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 6.7 area 535 850 877 325 1150 921 605 5263 11706 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 738.3 12.0 0.631 94.1 0.230 1 850 13.8 0.726 108.4 0.276 1 1026 16.7 0.877 130.8 0.320 1 325 5.3 0.278 41.4 0.110 1.08 1242 20.2 1.061 158.4 0.388 1 1363 22.2 1.164 173.8 0.425 1 605 9.8 0.517 77.1 0.196 6149 100 5 784 1.9 1 11706 11706 3:7-20 weight feces 6.5 mg a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso oppervlakte 290 571 462 258 300 422 response factor 1.38 1 1 1 1.08 1 1 total 2303 nor-hyodeoxy 4214 1 oppervlakte 400.2 571 541 258 324 625 0 2718 4214 % 14.7 21.0 19.9 9.5 11.9 23.0 0.0 100 4214 g bile acid 0.950 1.355 1.283 0.612 0.769 1.482 0.000 ng bile acid/ mg faeces 146.1 208.5 197.3 94.2 118.3 228.0 0.0 nmol bile acid/ mg faeces 0.358 0.531 0.483 0.250 0.289 0.558 0.000 6 992 2.5 3:7-21 weight feces a-muricholate deoxycholate cholate lithocholate -muricholate w-muricholate hyodeo/urso total nor-hyodeoxy 7.2 area 1184 3208 2501 1182 3049 3105 1201 15430 32495 mg response area corrected % g bile acid ng bile acid/ nmol bile acid/ factor for response factor mg feces mg feces 1.38 1633.92 9.1 0.503 69.8 0.171 1 3208 17.8 0.987 137.1 0.349 1 2926 16.2 0.900 125.1 0.306 1 1182 6.6 0.364 50.5 0.134 1.08 3292.92 18.3 1.013 140.7 0.344 1 4595 25.5 1.414 196.4 0.481 1 1201 6.7 0.370 51.3 0.131 18039 100 6 771 1.9 1 32495 32495 a-muri deoxy cholate lithochol b-muri w-muri hyodeoxy/urso 408.6 392.6 408.6 376.6 408.6 408.6 392.6 04-12-2006 Page A80 of A172 Page A81 of A172 group 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 AVERAGE SD cage# 1,2 3,4 5 1,2 3,4 5 1,2 3,4 5 6 7,8 9,10 6 7,8 9,10 6 7,8 9,10 11,12,13 15 14,16,17 11,12,13 15 14,16,17 11,12,13 15 14,16,17 18 19 20,21 18 19 20,21 18 19 20,21 22 24,25 23,26,27 22 24,25 23,26,27 22 24,25 23,26,27 28,29 31 30,32 28,29 31 30,32 28,29 31 30,32 33,34,35 36,37 38,39 33,34,35 36,37 38,39 33,34,35 36,37 38,39 GC sample# 1:1-1 1:1-2 1:1-3 2:1-1 2:1-2 2:1-3 3:1-1 3:1-2 3:1-3 a-muricholate 0.189 0.113 0.120 0.319 0.160 0.217 0.112 0.120 0.172 deoxycholate 0.532 0.274 0.311 0.528 0.305 0.366 0.764 0.298 0.330 1:2-4 1:2-5 1:2-6 2:2-4 2:2-5 2:2-6 3:2-4 3:2-5 3:2-6 0.082 0.080 0.150 0.165 0.145 0.238 0.149 0.121 0.204 0.345 0.278 0.496 0.396 0.261 0.559 0.377 0.249 0.508 1:3-7 1:3-8 1:3-9 2:3-7 2:3-8 2:3-9 3:3-7 3:3-8 3:3-9 0.119 0.102 0.158 0.068 0.100 0.154 0.113 0.101 0.133 0.420 0.443 0.397 0.437 0.269 0.326 0.356 0.259 0.352 1:4-10 1:4-11 1:4-12 2:4-10 2:4-11 2:4-12 3:4-10 3:4-11 3:4-12 0.055 0.231 0.159 0.102 0.189 0.179 0.093 0.140 0.146 0.142 0.250 0.320 0.223 0.191 0.224 0.174 0.264 0.210 1:5-13 1:5-14 1:5-15 2:5-13 2:5-14 2:5-15 3:5-13 3:5-14 3:5-15 0.093 0.084 0.089 0.103 0.118 0.209 0.087 0.080 0.122 0.352 0.238 0.325 0.191 0.291 0.526 0.187 0.155 0.355 1:6-16 1:6-17 1:6-18 2:6-16 2:6-17 2:6-18 3:6-16 3:6-17 3:6-18 0.079 0.099 0.047 0.074 0.107 0.121 0.122 0.129 0.080 0.176 0.203 0.195 0.213 0.399 0.285 0.181 0.215 0.164 1:7-19 1:7-20 1:7-21 2:7-19 2:7-20 2:7-21 3:7-19 3:7-20 3:7-21 0.262 0.306 0.238 0.241 0.184 0.270 0.230 0.358 0.171 0.799 0.343 0.498 0.532 0.479 1.065 0.276 0.531 0.349 bile acids (mol/g feces) cholate lithocholate b-muricholate w-muricholate hyodeo/urso 0.174 0.272 0.697 1.012 0.076 0.227 0.405 0.407 1.435 0.046 0.179 0.413 0.406 1.280 0.057 0.184 0.268 0.639 0.553 0.000 0.317 0.339 0.558 1.557 0.025 0.411 0.096 0.645 1.229 0.029 0.129 0.141 0.797 0.976 0.039 0.161 0.297 0.376 1.059 0.038 0.227 0.176 0.628 1.038 0.069 bile acids total (mol/g feces) 2.95 2.91 2.77 2.49 3.26 2.99 2.96 2.35 2.64 0.255 0.247 0.366 1.162 0.062 2.52 0.252 0.322 0.821 0.944 0.024 2.72 0.275 0.566 0.467 1.734 0.059 3.75 0.229 0.298 0.440 1.219 0.000 2.75 0.182 0.330 0.723 1.118 0.068 2.83 0.292 0.467 0.474 1.717 0.064 3.81 0.255 0.209 0.623 1.370 0.067 3.05 0.200 0.196 0.735 0.883 0.076 2.46 0.356 0.368 0.538 1.569 0.093 3.64 0.122 0.507 0.621 1.513 0.052 3.36 0.193 0.520 0.476 1.446 0.063 3.24 0.234 0.593 0.695 1.219 0.000 3.30 0.290 0.523 0.599 1.375 0.089 3.38 0.231 0.373 0.390 1.084 0.046 2.49 0.286 0.066 0.643 0.907 0.000 2.38 0.167 0.242 0.539 0.891 0.047 2.35 0.158 0.279 0.380 0.960 0.038 2.18 0.208 0.347 0.681 1.049 0.037 2.81 0.129 0.164 0.145 0.674 0.094 1.40 0.131 0.141 0.364 0.806 0.000 1.92 0.200 0.373 0.372 1.099 0.000 2.52 0.298 0.315 0.216 0.664 0.000 1.82 0.165 0.215 0.444 0.902 0.081 2.19 0.329 0.264 0.418 0.926 0.051 2.39 0.155 0.146 0.231 0.627 0.046 1.47 0.084 0.155 0.469 0.765 0.057 1.93 0.336 0.182 0.465 0.776 0.043 2.16 0.066 0.392 0.444 0.703 0.035 2.09 0.262 0.318 0.329 0.633 0.045 1.91 0.171 0.416 0.129 0.960 0.000 2.09 0.096 0.481 0.385 0.503 0.089 1.85 0.274 0.314 0.443 0.734 0.065 2.24 0.212 0.450 0.314 1.484 0.073 3.27 0.113 0.214 0.279 0.463 0.043 1.39 0.185 0.128 0.320 0.466 0.084 1.42 0.135 0.219 0.303 0.980 0.049 2.16 0.219 0.159 0.210 0.490 0.033 1.37 0.173 0.202 0.149 0.573 0.042 1.44 0.325 0.187 0.211 0.673 0.052 1.69 0.218 0.162 0.226 0.459 0.059 1.41 0.102 0.209 0.076 0.431 0.000 1.32 0.244 0.198 0.256 0.780 0.052 1.94 0.271 0.106 0.221 0.393 0.038 1.33 0.154 0.080 0.136 0.705 0.035 1.45 0.154 0.143 0.133 0.352 0.000 1.03 0.641 0.157 0.630 0.669 0.000 3.16 0.352 0.121 0.262 0.533 0.102 2.02 0.609 0.237 0.325 0.455 0.072 2.43 0.167 0.097 0.475 0.463 0.000 1.97 0.259 0.063 0.331 0.679 0.104 2.10 0.414 0.127 0.509 0.882 0.047 3.31 0.320 0.110 0.388 0.425 0.196 1.95 0.483 0.250 0.289 0.558 0.000 2.47 0.306 0.134 0.344 0.481 0.131 1.92 total feces (g) 3.075 2.4897 2.2946 4.692 3.7789 3.4106 3.5754 2.8781 2.7589 2.7619 4.0581 2.0935 4.075 6.371 3.4074 2.9279 4.3985 2.4575 3.4597 2.2994 2.3454 5.8147 3.7718 3.7727 4.025 2.5265 2.6905 1.689 3.1432 3.2545 2.7133 4.6407 5.2075 2.041 3.4464 3.9549 3.195 2.9408 2.6399 4.9712 4.4833 3.767 3.6113 3.3522 2.7034 2.5036 2.7083 2.3554 3.9684 3.9678 3.5666 2.9076 3.2449 2.7643 1.9697 2.0798 2.6705 3.4477 3.2646 4.3059 2.5117 2.4369 3.4107 average weight days (g) 123.6 2 87.25 2 88.55 2 122.45 3 86.55 3 88.2 3 121.65 2 86.55 2 88.05 2 94.55 2 126.6 2 82.2 2 94.25 3 126.45 3 82.1 3 94 2 126.6 2 81.85 2 118 2 83.55 2 82.8 2 118.75 3 83.5 3 83.05 3 119.6 2 84.2 2 83.45 2 59.5 2 116.4 2 112.2 2 58.85 3 116.55 3 111.85 3 58.7 2 117.45 2 112.9 2 110.7 2 88.35 2 84.6 2 111.6 3 88.5 3 85.55 3 112.85 2 88.9 2 85.5 2 87.1 2 112.8 2 85.4 2 87.15 3 112.45 3 84.55 3 87.65 2 111.95 2 84.65 2 81.35 2 82.25 2 101.05 2 82.35 3 82.75 3 101.95 3 83.5 2 83.35 2 103.2 2 total bile acid excretion (mol/100 gr mouse/day) 3.67 4.15 3.58 3.18 4.75 3.86 4.35 3.90 4.14 3.95 0.46 3.68 4.36 4.77 3.96 4.75 5.27 4.75 4.27 5.46 4.59 0.58 4.92 4.46 4.67 5.52 3.75 3.61 3.96 3.26 4.53 4.30 0.71 1.99 2.60 3.66 2.79 2.90 3.71 2.56 2.84 3.78 2.98 0.61 3.01 3.17 3.26 2.74 3.78 4.80 2.22 2.67 3.42 3.23 0.74 1.96 1.73 2.33 2.14 1.56 2.72 2.21 2.11 1.68 2.05 0.36 3.82 2.55 3.22 2.75 2.76 4.67 2.93 3.61 3.17 3.27 0.67 a-muricholate 6.39 3.87 4.33 12.82 4.92 7.24 3.78 5.13 6.53 6.11 2.80 3.26 2.93 4.00 6.00 5.13 6.26 4.87 4.91 5.61 4.77 1.16 3.56 3.16 4.81 2.02 4.03 6.46 4.81 4.64 4.75 4.25 1.26 3.90 12.03 6.31 5.60 8.64 7.48 6.31 7.24 6.76 7.14 2.26 4.47 4.39 4.25 5.55 5.26 6.39 6.31 5.63 5.63 5.32 0.80 5.80 6.86 2.76 5.24 8.11 6.24 9.16 8.87 7.83 6.76 2.02 8.29 15.16 9.78 12.23 8.76 8.15 11.84 14.48 8.92 10.84 2.69 deoxycholate 18.03 9.42 11.24 21.19 9.34 12.23 25.85 12.68 12.51 14.72 5.71 13.69 10.23 13.24 14.43 9.25 14.67 12.36 10.11 13.98 12.44 2.06 12.51 13.66 12.04 12.93 10.81 13.71 15.12 11.92 12.53 12.80 1.25 10.10 13.02 12.67 12.29 8.72 9.37 11.81 13.64 9.72 11.26 1.80 16.89 12.46 15.55 10.36 13.00 16.10 13.48 10.94 16.43 13.91 2.43 12.88 14.11 11.55 15.10 30.13 14.70 13.56 14.77 15.99 15.87 5.51 25.31 16.99 20.45 26.94 22.83 32.13 14.19 21.50 18.23 22.06 5.50 cholate 5.89 7.80 6.46 7.37 9.71 13.75 4.37 6.84 8.60 7.87 2.69 10.13 9.26 7.35 8.33 6.43 7.65 8.36 8.15 9.78 8.38 1.19 3.65 5.94 7.09 8.59 9.26 12.02 7.09 7.26 7.41 7.59 2.30 9.17 6.80 7.94 16.39 7.54 13.77 10.54 4.34 15.55 10.23 4.17 3.16 13.73 8.19 5.20 12.24 6.48 8.13 13.05 6.25 8.49 3.72 16.03 12.00 19.22 15.45 7.71 12.61 20.37 10.62 14.98 14.33 4.04 20.29 17.44 25.03 8.45 12.35 12.49 16.45 19.56 15.97 16.45 4.95 % of bile acids lithocholate b-muricholate 9.21 23.61 13.95 14.00 14.94 14.68 10.75 25.66 10.41 17.10 3.22 21.54 4.76 26.94 12.63 16.00 6.66 23.78 9.61 20.37 4.05 4.97 9.80 14.53 11.84 30.17 15.11 12.46 10.85 16.02 11.66 25.58 12.26 12.44 6.86 20.43 7.98 29.88 10.12 14.78 10.72 19.59 2.43 7.23 15.11 18.52 16.04 14.66 18.00 21.08 15.48 17.72 14.97 15.63 2.76 26.98 10.27 22.89 12.84 17.46 12.37 24.26 13.09 19.91 4.49 4.15 11.72 10.33 7.33 18.93 14.79 14.73 17.31 11.88 9.82 20.30 11.03 17.48 9.93 15.70 8.02 24.27 8.45 21.54 10.94 17.24 3.28 4.55 18.81 21.29 16.67 17.23 19.89 6.19 26.01 20.84 14.01 19.79 13.76 9.62 15.46 20.14 9.05 22.53 10.13 14.00 15.97 16.85 5.19 5.72 11.60 15.37 14.03 10.33 11.08 12.49 11.47 16.04 15.80 5.73 10.23 13.24 7.95 16.62 5.49 9.38 13.95 12.98 11.29 12.46 3.18 3.51 4.97 19.97 5.98 12.98 9.75 13.35 4.90 24.06 3.02 15.75 3.83 15.35 5.65 19.92 10.13 11.72 7.00 17.97 6.14 16.79 2.45 4.02 w-muricholate 34.30 49.37 46.29 22.20 47.75 41.06 32.99 45.11 39.32 39.82 8.75 46.13 34.70 46.28 44.38 39.57 45.04 44.92 35.90 43.16 42.23 4.42 45.10 44.58 36.99 40.65 43.46 38.07 37.82 44.13 37.37 40.91 3.42 48.08 41.89 43.55 36.53 41.25 38.74 42.61 39.53 35.96 40.90 3.75 33.71 33.19 45.94 27.22 32.79 45.43 33.36 32.84 45.31 36.64 6.96 35.89 39.79 39.81 32.54 32.51 40.27 29.53 48.49 34.26 37.01 5.73 21.18 26.38 18.69 23.43 32.35 26.62 21.86 22.60 25.08 24.24 3.97 hyodeo/urso 2.57 1.58 2.06 0.00 0.78 0.97 1.32 1.61 2.60 1.50 0.85 2.46 0.87 1.57 0.00 2.39 1.67 2.19 3.08 2.56 1.86 0.95 1.56 1.96 0.00 2.62 1.85 0.00 2.00 1.75 1.31 1.45 0.90 6.70 0.00 0.00 0.00 3.72 2.14 3.10 2.96 2.01 2.29 2.19 1.67 2.34 0.00 4.82 2.90 2.23 3.13 5.95 2.25 2.81 1.74 2.42 2.88 3.09 4.16 0.00 2.70 2.82 2.38 0.00 2.27 1.39 0.00 5.07 2.94 0.00 4.95 1.43 10.10 0.00 6.82 3.48 3.54 04-12-2006 Final version Appendix to TNO-report of 3M-02 Appendix to TNO-report of 3M-02 Final version 04-12-2006 bile acid total bile acids a-muricholate deoxycholate cholate lithocholate b-muricholate w-muricholate hyodeo/urso bile acid total bile acids a-muricholate deoxycholate cholate lithocholate b-muricholate w-muricholate hyodeo/urso bile acid total bile acids a-muricholate deoxycholate cholate lithocholate b-muricholate w-muricholate hyodeo/urso group 1 vs 2 nn p 9 9 0.019 9 9 0.258 9 9 0.863 9 9 0.340 9 9 0.605 9 9 0.666 9 9 0.931 9 9 0.436 group 2 vs 4 nn p 9 9 <0.001 9 9 0.006 9 9 0.113 9 9 0.546 9 9 0.931 9 9 0.605 9 9 0.387 9 9 0.730 group 3 vs 7 nn p 9 9 0.008 9 9 <0.001 9 9 <0.001 9 9 <0.001 9 9 0.004 9 9 0.161 9 9 <0.001 9 9 0.387 group 1 vs 3 nn p 9 9 0.297 9 9 0.077 9 9 0.931 9 9 1.000 9 9 0.063 9 9 1.000 9 9 0.730 9 9 1.000 group 2 vs 5 nn p 9 9 0.003 9 9 0.297 9 9 0.222 9 9 0.666 9 9 0.019 9 9 0.666 9 9 0.094 9 9 0.258 group 4 vs 5 nn p 9 9 0.489 9 9 0.031 9 9 0.031 9 9 0.340 9 9 0.031 9 9 1.000 9 9 0.161 9 9 0.546 Bile acid excretion, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 nn p nn p nn p 9 9 0.003 9 9 0.019 9 9 <0.001 9 9 0.190 9 9 0.863 9 9 0.258 9 9 0.340 9 9 0.796 9 9 0.258 9 9 0.190 9 9 0.931 9 9 0.002 9 9 0.666 9 9 0.019 9 9 0.340 9 9 0.258 9 9 0.190 9 9 0.003 9 9 1.000 9 9 0.258 9 9 0.297 9 9 0.387 9 9 0.040 9 9 0.094 group 2 vs 6 group 2 vs 7 group 3 vs 4 nn p nn p nn p 9 9 <0.001 9 9 0.001 9 9 0.002 9 9 0.024 9 9 <0.001 9 9 0.004 9 9 0.040 9 9 <0.001 9 9 0.094 9 9 0.001 9 9 <0.001 9 9 0.161 9 9 0.730 9 9 0.002 9 9 0.094 9 9 0.077 9 9 0.546 9 9 0.387 9 9 0.063 9 9 <0.001 9 9 0.863 9 9 0.258 9 9 0.546 9 9 0.222 group 4 vs 6 group 4 vs 7 group 5 vs 6 nn p nn p nn p 9 9 0.002 9 9 0.489 9 9 <0.001 9 9 1.000 9 9 0.003 9 9 0.063 9 9 0.003 9 9 <0.001 9 9 0.730 9 9 0.077 9 9 0.011 9 9 0.014 9 9 0.666 9 9 0.003 9 9 0.077 9 9 0.040 9 9 0.796 9 9 0.050 9 9 0.094 9 9 <0.001 9 9 0.931 9 9 0.931 9 9 0.605 9 9 0.931 group 1 vs 7 nn p 9 9 0.019 9 9 0.002 9 9 0.014 9 9 0.001 9 9 0.077 9 9 0.136 9 9 0.001 9 9 0.387 group 3 vs 5 nn p 9 9 0.008 9 9 0.094 9 9 0.340 9 9 0.796 9 9 0.387 9 9 0.340 9 9 0.258 9 9 0.031 group 5 vs 7 nn p 9 9 0.863 9 9 <0.001 9 9 <0.001 9 9 0.002 9 9 <0.001 9 9 0.666 9 9 <0.001 9 9 0.863 group 2 vs 3 nn p 9 9 0.387 9 9 0.258 9 9 1.000 9 9 0.222 9 9 0.040 9 9 0.605 9 9 0.436 9 9 0.340 group 3 vs 6 nn p 9 9 <0.001 9 9 0.008 9 9 0.063 9 9 0.001 9 9 0.161 9 9 0.001 9 9 0.094 9 9 0.050 group 6 vs 7 nn p 9 9 <0.001 9 9 0.002 9 9 0.011 9 9 0.340 9 9 0.002 9 9 0.050 9 9 <0.001 9 9 0.546 p<0.05 Page A82 of A172 Appendix to TNO-report of 3M-02 Final version E3. Fecal sterol measurements 1:1-1 weight feces 6.2 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 284344 0.98 278657 85.2 60.3 9718 25.13 cholestanol 9225 0.91 8395 2.6 1.8 293 0.75 lathosterol 10102 1 10102 3.1 2.2 352 0.91 campesterol 4671 1.56 7287 2.2 1.6 254 0.63 stigmasterol 1682 1.49 2506 0.8 0.5 87 0.21 b-sitosterol 13713 1.47 20158 6.2 4.4 703 1.70 b-sitostanol 1 0 0.0 0.0 0 0.00 total 323737 327105 100 71 11407 29.3 cholestane 111178 1.04 1:1-2 weight feces 6.5 mg 115625 115625 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 313994 0.98 307714 86.6 76.7 11803 30.52 cholestanol 9355 0.91 8513 2.4 2.1 327 0.84 lathosterol 9015 1 9015 2.5 2.2 346 0.89 campesterol 4796 1.56 7482 2.1 1.9 287 0.72 stigmasterol 1808 1.49 2694 0.8 0.7 103 0.25 b-sitosterol 13562 1.47 19936 5.6 5.0 765 1.84 b-sitostanol 1 0 0.0 0.0 0 0.00 total 352530 355354 100 89 13631 35.1 cholestane 96413 1.04 1:1-3 weight feces 7.1 mg 100270 100270 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 298829 0.98 292852 86.7 73.5 10347 26.76 cholestanol 8558 0.91 7788 2.3 2.0 275 0.71 lathosterol 10265 1 10265 3.0 2.6 363 0.94 campesterol 4356 1.56 6795 2.0 1.7 240 0.60 stigmasterol 1589 1.49 2368 0.7 0.6 84 0.20 b-sitosterol 12044 1.47 17705 5.2 4.4 626 1.51 b-sitostanol 1 0 0.0 0.0 0 0.00 total 335641 337773 100 85 11934 30.7 cholestane 95828 1.04 99661 99661 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A83 of A172 Appendix to TNO-report of 3M-02 Final version 2:1-1 weight feces 7 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 319680 0.98 313286 86.1 87.7 12532 32.41 cholestanol 11436 0.91 10407 2.9 2.9 416 1.07 lathosterol 10467 1 10467 2.9 2.9 419 1.08 campesterol 5023 1.56 7836 2.2 2.2 313 0.78 stigmasterol 1268 1.49 1889 0.5 0.5 76 0.18 b-sitosterol 13626 1.47 20030 5.5 5.6 801 1.93 b-sitostanol 1 0 0.0 0.0 0 0.00 total 361500 363916 100 102 14557 37.5 cholestane 85850 1.04 2:1-2 weight feces 10.4 mg 89284 89284 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 508256 0.98 498091 87.4 131.0 12591 32.56 cholestanol 15011 0.91 13660 2.4 3.6 345 0.89 lathosterol 13004 1 13004 2.3 3.4 329 0.85 campesterol 7404 1.56 11550 2.0 3.0 292 0.73 stigmasterol 2781 1.49 4144 0.7 1.1 105 0.25 b-sitosterol 19988 1.47 29382 5.2 7.7 743 1.79 b-sitostanol 1 0 0.0 0.0 0 0.00 total 566444 569831 100 150 14405 37.1 cholestane 91434 1.04 2:1-3 weight feces 6.8 mg 95091 95091 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 292666 0.98 286813 86.8 89.9 13215 34.17 cholestanol 8651 0.91 7872 2.4 2.5 363 0.93 lathosterol 9600 1 9600 2.9 3.0 442 1.14 campesterol 4193 1.56 6541 2.0 2.0 301 0.75 stigmasterol 1436 1.49 2140 0.6 0.7 99 0.24 b-sitosterol 11876 1.47 17458 5.3 5.5 804 1.94 b-sitostanol 1 0 0.0 0.0 0 0.00 total 328422 330424 100 104 15225 39.2 cholestane 76721 1.04 79790 79790 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A84 of A172 Appendix to TNO-report of 3M-02 Final version 3:1-1 weight feces 6.1 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 440429 0.98 431620 87.6 73.9 12121 31.34 cholestanol 13442 0.91 12232 2.5 2.1 344 0.88 lathosterol 11803 1 11803 2.4 2.0 331 0.86 campesterol 6241 1.56 9736 2.0 1.7 273 0.68 stigmasterol 2114 1.49 3150 0.6 0.5 88 0.21 b-sitosterol 16460 1.47 24196 4.9 4.1 679 1.64 b-sitostanol 1 0 0.0 0.0 0 0.00 total 490489 492738 100 84 13837 35.6 cholestane 140330 1.04 3:1-2 weight feces 5.3 mg 145943 145943 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 237491 0.98 232741 86.7 55.6 10481 27.10 cholestanol 7441 0.91 6771 2.5 1.6 305 0.78 lathosterol 6444 1 6444 2.4 1.5 290 0.75 campesterol 3777 1.56 5892 2.2 1.4 265 0.66 stigmasterol 982 1.49 1463 0.5 0.3 66 0.16 b-sitosterol 10254 1.47 15073 5.6 3.6 679 1.64 b-sitostanol 1 0 0.0 0.0 0 0.00 total 266389 268385 100 64 12086 31.1 cholestane 100714 1.04 3:1-3 weight feces 6.7 mg 104743 104743 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 404372 0.98 396285 88.1 72.6 10838 28.03 cholestanol 11102 0.91 10103 2.2 1.9 276 0.71 lathosterol 11373 1 11373 2.5 2.1 311 0.80 campesterol 5436 1.56 8480 1.9 1.6 232 0.58 stigmasterol 2003 1.49 2984 0.7 0.5 82 0.20 b-sitosterol 14146 1.47 20795 4.6 3.8 569 1.37 b-sitostanol 1 0 0.0 0.0 0 0.00 total 448432 450020 100 82 12307 31.7 cholestane 131190 1.04 136438 136438 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A85 of A172 Appendix to TNO-report of 3M-02 Final version 1:2-4 weight feces 9.1 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 473290 0.98 463824 88.5 123.6 13579 35.12 cholestanol 9709 0.91 8835 1.7 2.4 259 0.67 lathosterol 12754 1 12754 2.4 3.4 373 0.97 campesterol 6031 1.56 9408 1.8 2.5 275 0.69 stigmasterol 2406 1.49 3585 0.7 1.0 105 0.25 b-sitosterol 17298 1.47 25428 4.9 6.8 744 1.80 b-sitostanol 1 0 0.0 0.0 0 0.00 total 521488 523835 100 140 15336 39.5 cholestane 90227 1.04 1:2-5 weight feces 7.2 mg 93836 93836 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 446804 0.98 437868 90.1 114.1 15846 40.98 cholestanol 6124 0.91 5573 1.1 1.5 202 0.52 lathosterol 14209 1 14209 2.9 3.7 514 1.33 campesterol 4560 1.56 7114 1.5 1.9 257 0.64 stigmasterol 1811 1.49 2698 0.6 0.7 98 0.24 b-sitosterol 12767 1.47 18767 3.9 4.9 679 1.64 b-sitostanol 1 0 0.0 0.0 0 0.00 total 486275 486229 100 127 17596 45.3 cholestane 92255 1.04 1:2-6 weight feces 6.2 mg 95945 95945 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 298782 0.98 292806 84.8 65.1 10502 27.16 cholestanol 11596 0.91 10552 3.1 2.3 378 0.97 lathosterol 8473 1 8473 2.5 1.9 304 0.79 campesterol 5240 1.56 8174 2.4 1.8 293 0.73 stigmasterol 1956 1.49 2914 0.8 0.6 105 0.25 b-sitosterol 15108 1.47 22209 6.4 4.9 797 1.92 b-sitostanol 1 0 0.0 0.0 0 0.00 total 341155 345129 100 77 12379 31.8 cholestane 108095 1.04 112419 112419 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A86 of A172 Appendix to TNO-report of 3M-02 Final version 2:2-4 weight feces 8.6 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 481673 0.98 472040 88.7 118.2 13750 35.56 cholestanol 11693 0.91 10641 2.0 2.7 310 0.80 lathosterol 12978 1 12978 2.4 3.3 378 0.98 campesterol 6203 1.56 9677 1.8 2.4 282 0.70 stigmasterol 1175 1.49 1751 0.3 0.4 51 0.12 b-sitosterol 17089 1.47 25121 4.7 6.3 732 1.76 b-sitostanol 1 0 0.0 0.0 0 0.00 total 530811 532206 100 133 15502 39.9 cholestane 95962 1.04 2:2-5 weight feces 9.9 mg 99800 99800 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 434117 0.98 425435 88.8 152.2 15373 39.75 cholestanol 8812 0.91 8019 1.7 2.9 290 0.75 lathosterol 13243 1 13243 2.8 4.7 479 1.24 campesterol 5232 1.56 8162 1.7 2.9 295 0.74 stigmasterol 2068 1.49 3081 0.6 1.1 111 0.27 b-sitosterol 14440 1.47 21227 4.4 7.6 767 1.85 b-sitostanol 1 0 0.0 0.0 0 0.00 total 477912 479167 100 171 17315 44.6 cholestane 67196 1.04 2:2-6 weight feces 6 mg 69884 69884 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 268071 0.98 262710 85.9 72.3 12043 31.14 cholestanol 10152 0.91 9238 3.0 2.5 423 1.09 lathosterol 6784 1 6784 2.2 1.9 311 0.80 campesterol 4476 1.56 6983 2.3 1.9 320 0.80 stigmasterol 1045 1.49 1557 0.5 0.4 71 0.17 b-sitosterol 12736 1.47 18722 6.1 5.1 858 2.07 b-sitostanol 1 0 0.0 0.0 0 0.00 total 303264 305993 100 84 14027 36.1 cholestane 87400 1.04 90896 90896 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A87 of A172 Appendix to TNO-report of 3M-02 Final version 3:2-4 weight feces 5.6 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 269271 0.98 263886 87.6 66.6 11896 30.76 cholestanol 7132 0.91 6490 2.2 1.6 293 0.75 lathosterol 7058 1 7058 2.3 1.8 318 0.82 campesterol 3976 1.56 6203 2.1 1.6 280 0.70 stigmasterol 1401 1.49 2087 0.7 0.5 94 0.23 b-sitosterol 10601 1.47 15583 5.2 3.9 702 1.69 b-sitostanol 1 0 0.0 0.0 0 0.00 total 299439 301307 100 76 13582 35.0 cholestane 95225 1.04 3:2-5 weight feces 6.9 mg 99034 99034 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 398275 0.98 390310 89.5 79.9 11581 29.95 cholestanol 7905 0.91 7194 1.6 1.5 213 0.55 lathosterol 11474 1 11474 2.6 2.3 340 0.88 campesterol 4821 1.56 7521 1.7 1.5 223 0.56 stigmasterol 1507 1.49 2245 0.5 0.5 67 0.16 b-sitosterol 11865 1.47 17442 4.0 3.6 518 1.25 b-sitostanol 1 0 0.0 0.0 0 0.00 total 435847 436185 100 89 12942 33.3 cholestane 117414 1.04 3:2-6 weight feces 7.2 mg 122111 122111 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 336997 0.98 330257 86.2 80.7 11206 28.98 cholestanol 12422 0.91 11304 2.9 2.8 384 0.99 lathosterol 7963 1 7963 2.1 1.9 270 0.70 campesterol 5484 1.56 8555 2.2 2.1 290 0.72 stigmasterol 2009 1.49 2993 0.8 0.7 102 0.25 b-sitosterol 15075 1.47 22160 5.8 5.4 752 1.81 b-sitostanol 1 0 0.0 0.0 0 0.00 total 379950 383233 100 94 13003 33.4 cholestane 98396 1.04 102332 102332 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A88 of A172 Appendix to TNO-report of 3M-02 Final version 1:3-7 weight feces 6 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 240788 0.98 235972 85.5 67.4 11229 29.04 cholestanol 7267 0.91 6613 2.4 1.9 315 0.81 lathosterol 9283 1 9283 3.4 2.7 442 1.14 campesterol 3808 1.56 5940 2.2 1.7 283 0.71 stigmasterol 1488 1.49 2217 0.8 0.6 106 0.26 b-sitosterol 10778 1.47 15844 5.7 4.5 754 1.82 b-sitostanol 1 0 0.0 0.0 0 0.00 total 273412 275869 100 79 13128 33.8 cholestane 84191 1.04 1:3-8 weight feces 7 mg 87559 87559 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 283543 0.98 277872 85.5 71.7 10245 26.49 cholestanol 10969 0.91 9982 3.1 2.6 368 0.95 lathosterol 8285 1 8285 2.5 2.1 305 0.79 campesterol 4663 1.56 7274 2.2 1.9 268 0.67 stigmasterol 1774 1.49 2643 0.8 0.7 97 0.24 b-sitosterol 12953 1.47 19041 5.9 4.9 702 1.69 b-sitostanol 1 0 0.0 0.0 0 0.00 total 322187 325097 100 84 11986 30.8 cholestane 93139 1.04 1:3-9 weight feces 10.8 mg 96865 96865 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 529012 0.98 518432 89.0 153.4 14205 36.73 cholestanol 14038 0.91 12775 2.2 3.8 350 0.90 lathosterol 11702 1 11702 2.0 3.5 321 0.83 campesterol 6647 1.56 10369 1.8 3.1 284 0.71 stigmasterol 1317 1.49 1962 0.3 0.6 54 0.13 b-sitosterol 18688 1.47 27471 4.7 8.1 753 1.82 b-sitostanol 1 0 0.0 0.0 0 0.00 total 581404 582711 100 172 15966 41.1 cholestane 81233 1.04 84482 84482 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A89 of A172 Appendix to TNO-report of 3M-02 Final version 2:3-7 weight feces 8 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 330311 0.98 323705 86.2 95.0 11875 30.71 cholestanol 9369 0.91 8526 2.3 2.5 313 0.80 lathosterol 11675 1 11675 3.1 3.4 428 1.11 campesterol 5157 1.56 8045 2.1 2.4 295 0.74 stigmasterol 1936 1.49 2885 0.8 0.8 106 0.26 b-sitosterol 14114 1.47 20748 5.5 6.1 761 1.84 b-sitostanol 1 0 0.0 0.0 0 0.00 total 372562 375583 100 110 13778 35.4 cholestane 81910 1.04 2:3-8 weight feces 7.3 mg 85186 85186 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 361988 0.98 354748 87.7 89.9 12322 31.86 cholestanol 10955 0.91 9969 2.5 2.5 346 0.89 lathosterol 10492 1 10492 2.6 2.7 364 0.94 campesterol 4873 1.56 7602 1.9 1.9 264 0.66 stigmasterol 1595 1.49 2377 0.6 0.6 83 0.20 b-sitosterol 13274 1.47 19513 4.8 4.9 678 1.63 b-sitostanol 1 0 0.0 0.0 0 0.00 total 403177 404701 100 103 14057 36.2 cholestane 94804 1.04 2:3-9 weight feces 7.7 mg 98596 98596 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 409014 0.98 400834 88.8 109.4 14211 36.75 cholestanol 11289 0.91 10273 2.3 2.8 364 0.94 lathosterol 9106 1 9106 2.0 2.5 323 0.83 campesterol 5174 1.56 8071 1.8 2.2 286 0.71 stigmasterol 1042 1.49 1553 0.3 0.4 55 0.13 b-sitosterol 14716 1.47 21633 4.8 5.9 767 1.85 b-sitostanol 1 0 0.0 0.0 0 0.00 total 450341 451469 100 123 16006 41.2 cholestane 88054 1.04 91576 91576 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A90 of A172 Appendix to TNO-report of 3M-02 Final version 3:3-7 weight feces 8.2 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 428635 0.98 420062 87.0 95.1 11598 29.99 cholestanol 12166 0.91 11071 2.3 2.5 306 0.79 lathosterol 13406 1 13406 2.8 3.0 370 0.96 campesterol 6496 1.56 10134 2.1 2.3 280 0.70 stigmasterol 2441 1.49 3637 0.8 0.8 100 0.24 b-sitosterol 16860 1.47 24784 5.1 5.6 684 1.65 b-sitostanol 1 0 0.0 0.0 0 0.00 total 480004 483094 100 109 13338 34.3 cholestane 106174 1.04 3:3-8 weight feces 7.6 mg 110421 110421 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 370433 0.98 363024 86.3 74.7 9828 25.41 cholestanol 13556 0.91 12336 2.9 2.5 334 0.86 lathosterol 9317 1 9317 2.2 1.9 252 0.65 campesterol 6050 1.56 9438 2.2 1.9 256 0.64 stigmasterol 2043 1.49 3044 0.7 0.6 82 0.20 b-sitosterol 15825 1.47 23263 5.5 4.8 630 1.52 b-sitostanol 1 0 0.0 0.0 0 0.00 total 417224 420422 100 87 11382 29.3 cholestane 116833 1.04 3:3-9 weight feces 6 mg 121506 121506 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 327522 0.98 320972 88.4 71.3 11877 30.71 cholestanol 9565 0.91 8704 2.4 1.9 322 0.83 lathosterol 7411 1 7411 2.0 1.6 274 0.71 campesterol 4458 1.56 6954 1.9 1.5 257 0.64 stigmasterol 1015 1.49 1512 0.4 0.3 56 0.14 b-sitosterol 11869 1.47 17447 4.8 3.9 646 1.56 b-sitostanol 1 0 0.0 0.0 0 0.00 total 361840 363001 100 81 13432 34.6 cholestane 108272 1.04 112603 112603 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A91 of A172 Appendix to TNO-report of 3M-02 Final version 1:4-10 weight feces 5.9 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 362101 0.98 354859 88.7 81.6 13829 35.76 cholestanol 9381 0.91 8537 2.1 2.0 333 0.86 lathosterol 7899 1 7899 2.0 1.8 308 0.80 campesterol 4622 1.56 7210 1.8 1.7 281 0.70 stigmasterol 1882 1.49 2804 0.7 0.6 109 0.26 b-sitosterol 12850 1.47 18890 4.7 4.3 736 1.78 b-sitostanol 1 0 0.0 0.0 0 0.00 total 398735 400199 100 92 15595 40.2 cholestane 104552 1.04 1:4-11 weight feces 8 mg 108734 108734 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 489702 0.98 479908 90.0 107.0 13380 34.60 cholestanol 15062 0.91 13706 2.6 3.1 382 0.98 lathosterol 10888 1 10888 2.0 2.4 304 0.78 campesterol 4869 1.56 7596 1.4 1.7 212 0.53 stigmasterol 748 1.49 1115 0.2 0.2 31 0.08 b-sitosterol 13552 1.47 19921 3.7 4.4 555 1.34 b-sitostanol 1 0 0.0 0.0 0 0.00 total 534821 533134 100 119 14864 38.3 cholestane 107776 1.04 1:4-12 weight feces 6.1 mg 112087 112087 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 459088 0.98 449906 89.5 96.7 15857 41.01 cholestanol 15039 0.91 13685 2.7 2.9 482 1.24 lathosterol 11182 1 11182 2.2 2.4 394 1.02 campesterol 4797 1.56 7483 1.5 1.6 264 0.66 stigmasterol 686 1.49 1022 0.2 0.2 36 0.09 b-sitosterol 13085 1.47 19235 3.8 4.1 678 1.63 b-sitostanol 1 0 0.0 0.0 0 0.00 total 503877 502514 100 108 17711 45.6 cholestane 111810 1.04 116282 116282 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A92 of A172 Appendix to TNO-report of 3M-02 Final version 2:4-10 weight feces 7.7 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 521217 0.98 510793 90.7 142.2 18473 47.77 cholestanol 14915 0.91 13573 2.4 3.8 491 1.26 lathosterol 11055 1 11055 2.0 3.1 400 1.03 campesterol 4779 1.56 7455 1.3 2.1 270 0.67 stigmasterol 743 1.49 1107 0.2 0.3 40 0.10 b-sitosterol 13109 1.47 19270 3.4 5.4 697 1.68 b-sitostanol 1 0 0.0 0.0 0 0.00 total 565818 563253 100 157 20370 52.5 cholestane 86322 1.04 2:4-11 weight feces 6.4 mg 89775 89775 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 497491 0.98 487541 90.8 118.8 18567 48.01 cholestanol 13351 0.91 12149 2.3 3.0 463 1.19 lathosterol 10137 1 10137 1.9 2.5 386 1.00 campesterol 4786 1.56 7466 1.4 1.8 284 0.71 stigmasterol 727 1.49 1083 0.2 0.3 41 0.10 b-sitosterol 12456 1.47 18310 3.4 4.5 697 1.68 b-sitostanol 1 0 0.0 0.0 0 0.00 total 538948 536687 100 131 20439 52.7 cholestane 98627 1.04 2:4-12 weight feces 8.9 mg 102572 102572 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 572090 0.98 560648 90.6 171.6 19286 49.87 cholestanol 15045 0.91 13691 2.2 4.2 471 1.21 lathosterol 12013 1 12013 1.9 3.7 413 1.07 campesterol 5734 1.56 8945 1.4 2.7 308 0.77 stigmasterol 676 1.49 1007 0.2 0.3 35 0.08 b-sitosterol 15286 1.47 22470 3.6 6.9 773 1.86 b-sitostanol 1 0 0.0 0.0 0 0.00 total 620844 618775 100 189 21285 54.9 cholestane 78519 1.04 81660 81660 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A93 of A172 Appendix to TNO-report of 3M-02 Final version 3:4-10 weight feces 6.8 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 434209 0.98 425525 90.4 89.6 13179 34.08 cholestanol 12431 0.91 11312 2.4 2.4 350 0.90 lathosterol 9196 1 9196 2.0 1.9 285 0.74 campesterol 4448 1.56 6939 1.5 1.5 215 0.54 stigmasterol 651 1.49 970 0.2 0.2 30 0.07 b-sitosterol 11421 1.47 16789 3.6 3.5 520 1.25 b-sitostanol 1 0 0.0 0.0 0 0.00 total 472356 470731 100 99 14579 37.6 cholestane 114142 1.04 3:4-11 weight feces 6.8 mg 118708 118708 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 690765 0.98 676950 91.4 122.7 18042 46.66 cholestanol 15497 0.91 14102 1.9 2.6 376 0.97 lathosterol 14645 1 14645 2.0 2.7 390 1.01 campesterol 6317 1.56 9855 1.3 1.8 263 0.66 stigmasterol 805 1.49 1199 0.2 0.2 32 0.08 b-sitosterol 16234 1.47 23864 3.2 4.3 636 1.53 b-sitostanol 1 0 0.0 0.0 0 0.00 total 744263 740615 100 134 19739 50.9 cholestane 132638 1.04 3:4-12 weight feces 7.7 mg 137944 137944 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 657017 0.98 643877 91.0 121.8 15814 40.89 cholestanol 16315 0.91 14847 2.1 2.8 365 0.94 lathosterol 13781 1 13781 1.9 2.6 338 0.88 campesterol 6401 1.56 9986 1.4 1.9 245 0.61 stigmasterol 812 1.49 1210 0.2 0.2 30 0.07 b-sitosterol 16291 1.47 23948 3.4 4.5 588 1.42 b-sitostanol 1 0 0.0 0.0 0 0.00 total 710617 707648 100 134 17380 44.8 cholestane 127109 1.04 132193 132193 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A94 of A172 Appendix to TNO-report of 3M-02 Final version 1:5-13 weight feces 7.8 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 482166 0.98 472523 89.1 129.2 16561 42.83 cholestanol 15148 0.91 13785 2.6 3.8 483 1.24 lathosterol 13001 1 13001 2.5 3.6 456 1.18 campesterol 5131 1.56 8004 1.5 2.2 281 0.70 stigmasterol 1060 1.49 1579 0.3 0.4 55 0.13 b-sitosterol 14512 1.47 21333 4.0 5.8 748 1.80 b-sitostanol 1 0 0.0 0.0 0 0.00 total 531018 530225 100 145 18583 47.9 cholestane 87934 1.04 1:5-14 weight feces 13.4 mg 91451 91451 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 634279 0.98 621593 89.9 187.2 13972 36.13 cholestanol 12599 0.91 11465 1.7 3.5 258 0.66 lathosterol 16167 1 16167 2.3 4.9 363 0.94 campesterol 6696 1.56 10446 1.5 3.1 235 0.59 stigmasterol 2927 1.49 4361 0.6 1.3 98 0.24 b-sitosterol 18799 1.47 27635 4.0 8.3 621 1.50 b-sitostanol 1 0 0.0 0.0 0 0.00 total 691467 691667 100 208 15547 40.1 cholestane 79811 1.04 1:5-15 weight feces 6.2 mg 83003 83003 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 255399 0.98 250291 85.5 66.1 10669 27.59 cholestanol 13187 0.91 12000 4.1 3.2 512 1.32 lathosterol 6307 1 6307 2.2 1.7 269 0.70 campesterol 3843 1.56 5995 2.0 1.6 256 0.64 stigmasterol 729 1.49 1086 0.4 0.3 46 0.11 b-sitosterol 11657 1.47 17136 5.9 4.5 730 1.76 b-sitostanol 1 0 0.0 0.0 0 0.00 total 291122 292815 100 77 12481 32.1 cholestane 90959 1.04 94597 94597 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A95 of A172 Appendix to TNO-report of 3M-02 Final version 2:5-13 weight feces 7.9 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 455921 0.98 446803 89.0 124.6 15774 40.79 cholestanol 13655 0.91 12426 2.5 3.5 439 1.13 lathosterol 12928 1 12928 2.6 3.6 456 1.18 campesterol 5203 1.56 8117 1.6 2.3 287 0.72 stigmasterol 898 1.49 1338 0.3 0.4 47 0.11 b-sitosterol 13941 1.47 20493 4.1 5.7 723 1.74 b-sitostanol 1 0 0.0 0.0 0 0.00 total 502546 502105 100 140 17726 45.7 cholestane 86190 1.04 2:5-14 weight feces 6.9 mg 89638 89638 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 302926 0.98 296867 90.2 101.4 14697 38.01 cholestanol 6271 0.91 5707 1.7 1.9 283 0.73 lathosterol 8314 1 8314 2.5 2.8 412 1.06 campesterol 3149 1.56 4912 1.5 1.7 243 0.61 stigmasterol 541 1.49 806 0.2 0.3 40 0.10 b-sitosterol 8475 1.47 12458 3.8 4.3 617 1.49 b-sitostanol 1 0 0.0 0.0 0 0.00 total 329676 329065 100 112 16291 42.0 cholestane 70369 1.04 2:5-15 weight feces 4.8 mg 73184 73184 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 210645 0.98 206432 83.6 52.2 10870 28.11 cholestanol 13493 0.91 12279 5.0 3.1 647 1.66 lathosterol 5461 1 5461 2.2 1.4 288 0.74 campesterol 3832 1.56 5978 2.4 1.5 315 0.79 stigmasterol 733 1.49 1092 0.4 0.3 58 0.14 b-sitosterol 10573 1.47 15542 6.3 3.9 818 1.97 b-sitostanol 1 0 0.0 0.0 0 0.00 total 244737 246784 100 62 12995 33.4 cholestane 95106 1.04 98910 98910 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A96 of A172 Appendix to TNO-report of 3M-02 Final version 3:5-13 weight feces 11.8 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 760655 0.98 745442 88.7 146.6 12425 32.13 cholestanol 21051 0.91 19156 2.3 3.8 319 0.82 lathosterol 19855 1 19855 2.4 3.9 331 0.86 campesterol 9056 1.56 14127 1.7 2.8 235 0.59 stigmasterol 3585 1.49 5342 0.6 1.1 89 0.22 b-sitosterol 24867 1.47 36554 4.3 7.2 609 1.47 b-sitostanol 1 0 0.0 0.0 0 0.00 total 839069 840477 100 165 14009 36.1 cholestane 122218 1.04 3:5-14 weight feces 7.2 mg 127107 127107 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 475539 0.98 466028 90.3 88.3 12257 31.70 cholestanol 8894 0.91 8094 1.6 1.5 213 0.55 lathosterol 11319 1 11319 2.2 2.1 298 0.77 campesterol 4936 1.56 7700 1.5 1.5 203 0.51 stigmasterol 2133 1.49 3178 0.6 0.6 84 0.20 b-sitosterol 13417 1.47 19723 3.8 3.7 519 1.25 b-sitostanol 1 0 0.0 0.0 0 0.00 total 516238 516042 100 98 13573 35.0 cholestane 126938 1.04 3:5-15 weight feces 7.1 mg 132016 132016 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 369324 0.98 361938 85.4 62.2 8763 22.66 cholestanol 21096 0.91 19197 4.5 3.3 465 1.20 lathosterol 8285 1 8285 2.0 1.4 201 0.52 campesterol 6116 1.56 9541 2.3 1.6 231 0.58 stigmasterol 1212 1.49 1806 0.4 0.3 44 0.11 b-sitosterol 15819 1.47 23254 5.5 4.0 563 1.36 b-sitostanol 1 0 0.0 0.0 0 0.00 total 421852 424021 100 73 10267 26.4 cholestane 139834 1.04 145427 145427 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A97 of A172 Appendix to TNO-report of 3M-02 Final version 1:6-16 weight feces 5.5 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 316760 0.98 310425 88.4 75.3 13682 35.38 cholestanol 10679 0.91 9718 2.8 2.4 428 1.10 lathosterol 7883 1 7883 2.2 1.9 347 0.90 campesterol 3863 1.56 6026 1.7 1.5 266 0.66 stigmasterol 573 1.49 854 0.2 0.2 38 0.09 b-sitosterol 10951 1.47 16098 4.6 3.9 710 1.71 b-sitostanol 1 0 0.0 0.0 0 0.00 total 350709 351004 100 85 15471 39.8 cholestane 99161 1.04 1:6-17 weight feces 5.1 mg 103127 103127 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 283717 0.98 278043 86.7 61.2 12006 31.05 cholestanol 12638 0.91 11501 3.6 2.5 497 1.28 lathosterol 7544 1 7544 2.4 1.7 326 0.84 campesterol 4033 1.56 6291 2.0 1.4 272 0.68 stigmasterol 593 1.49 884 0.3 0.2 38 0.09 b-sitosterol 11162 1.47 16408 5.1 3.6 709 1.71 b-sitostanol 1 0 0.0 0.0 0 0.00 total 319687 320670 100 71 13847 35.6 cholestane 109156 1.04 1:6-18 weight feces 9.3 mg 113522 113522 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 432750 0.98 424095 87.8 121.3 13048 33.74 cholestanol 15694 0.91 14282 3.0 4.1 439 1.13 lathosterol 11216 1 11216 2.3 3.2 345 0.89 campesterol 5531 1.56 8628 1.8 2.5 265 0.66 stigmasterol 828 1.49 1234 0.3 0.4 38 0.09 b-sitosterol 15866 1.47 23323 4.8 6.7 718 1.73 b-sitostanol 1 0 0.0 0.0 0 0.00 total 481885 482778 100 138 14853 38.2 cholestane 84015 1.04 87376 87376 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A98 of A172 Appendix to TNO-report of 3M-02 Final version 2:6-16 weight feces 5.2 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 306320 0.98 300194 89.1 71.3 13705 35.44 cholestanol 10414 0.91 9477 2.8 2.2 433 1.11 lathosterol 7025 1 7025 2.1 1.7 321 0.83 campesterol 3400 1.56 5304 1.6 1.3 242 0.60 stigmasterol 583 1.49 869 0.3 0.2 40 0.10 b-sitosterol 9578 1.47 14080 4.2 3.3 643 1.55 b-sitostanol 1 0 0.0 0.0 0 0.00 total 337320 336948 100 80 15383 39.6 cholestane 101256 1.04 2:6-17 weight feces 4.6 mg 105306 105306 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 264813 0.98 259517 86.0 58.0 12612 32.61 cholestanol 13978 0.91 12720 4.2 2.8 618 1.59 lathosterol 6351 1 6351 2.1 1.4 309 0.80 campesterol 4076 1.56 6359 2.1 1.4 309 0.77 stigmasterol 392 1.49 584 0.2 0.1 28 0.07 b-sitosterol 11081 1.47 16289 5.4 3.6 792 1.91 b-sitostanol 1 0 0.0 0.0 0 0.00 total 300691 301819 100 67 14667 37.8 cholestane 107534 1.04 2:6-18 weight feces 7.1 mg 111835 111835 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 415098 0.98 406796 88.0 89.1 12553 32.46 cholestanol 15248 0.91 13876 3.0 3.0 428 1.10 lathosterol 11257 1 11257 2.4 2.5 347 0.90 campesterol 5140 1.56 8018 1.7 1.8 247 0.62 stigmasterol 790 1.49 1177 0.3 0.3 36 0.09 b-sitosterol 14501 1.47 21316 4.6 4.7 658 1.59 b-sitostanol 1 0 0.0 0.0 0 0.00 total 462034 462441 100 101 14270 36.8 cholestane 109718 1.04 114107 114107 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A99 of A172 Appendix to TNO-report of 3M-02 Final version 3:6-16 weight feces 9.4 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 528030 0.98 517469 90.2 120.9 12861 33.26 cholestanol 15892 0.91 14462 2.5 3.4 359 0.92 lathosterol 10925 1 10925 1.9 2.6 272 0.70 campesterol 5503 1.56 8585 1.5 2.0 213 0.53 stigmasterol 761 1.49 1134 0.2 0.3 28 0.07 b-sitosterol 14410 1.47 21183 3.7 4.9 526 1.27 b-sitostanol 1 0 0.0 0.0 0 0.00 total 575521 573757 100 134 14260 36.8 cholestane 102895 1.04 3:6-17 weight feces 5.2 mg 107011 107011 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 325132 0.98 318629 87.3 55.5 10671 27.59 cholestanol 15757 0.91 14339 3.9 2.5 480 1.24 lathosterol 7236 1 7236 2.0 1.3 242 0.63 campesterol 4495 1.56 7012 1.9 1.2 235 0.59 stigmasterol 611 1.49 910 0.2 0.2 30 0.07 b-sitosterol 11479 1.47 16874 4.6 2.9 565 1.36 b-sitostanol 1 0 0.0 0.0 0 0.00 total 364710 365001 100 64 12224 31.5 cholestane 138038 1.04 3:6-18 weight feces 6.9 mg 143560 143560 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 716301 0.98 701975 88.7 96.4 13976 36.14 cholestanol 24577 0.91 22365 2.8 3.1 445 1.15 lathosterol 18733 1 18733 2.4 2.6 373 0.96 campesterol 8508 1.56 13272 1.7 1.8 264 0.66 stigmasterol 1102 1.49 1642 0.2 0.2 33 0.08 b-sitosterol 22755 1.47 33450 4.2 4.6 666 1.61 b-sitostanol 1 0 0.0 0.0 0 0.00 total 791976 791437 100 109 15758 40.6 cholestane 174978 1.04 181977 181977 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A100 of A172 Appendix to TNO-report of 3M-02 Final version 1:7-19 weight feces 7.5 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 449806 0.98 440810 86.5 98.4 13115 33.92 cholestanol 25289 0.91 23013 4.5 5.1 685 1.76 lathosterol 7667 1 7667 1.5 1.7 228 0.59 campesterol 6477 1.56 10104 2.0 2.3 301 0.75 stigmasterol 917 1.49 1366 0.3 0.3 41 0.10 b-sitosterol 18151 1.47 26682 5.2 6.0 794 1.91 b-sitostanol 1 0 0.0 0.0 0 0.00 total 508307 509642 100 114 15163 39.0 cholestane 107726 1.04 1:7-20 weight feces 10.8 mg 112035 112035 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 497481 0.98 487531 85.5 119.2 11041 28.55 cholestanol 30777 0.91 28007 4.9 6.9 634 1.63 lathosterol 9362 1 9362 1.6 2.3 212 0.55 campesterol 7714 1.56 12034 2.1 2.9 273 0.68 stigmasterol 1041 1.49 1551 0.3 0.4 35 0.09 b-sitosterol 21441 1.47 31518 5.5 7.7 714 1.72 b-sitostanol 1 0 0.0 0.0 0 0.00 total 567816 570004 100 139 12909 33.2 cholestane 98281 1.04 1:7-21 weight feces 8.5 mg 102212 102212 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 328489 0.98 321919 86.3 100.2 11792 30.49 cholestanol 18232 0.91 16591 4.4 5.2 608 1.56 lathosterol 5881 1 5881 1.6 1.8 215 0.56 campesterol 5028 1.56 7844 2.1 2.4 287 0.72 stigmasterol 654 1.49 974 0.3 0.3 36 0.09 b-sitosterol 13624 1.47 20027 5.4 6.2 734 1.77 b-sitostanol 1 0 0.0 0.0 0 0.00 total 371908 373237 100 116 13671 35.2 cholestane 77207 1.04 80295 80295 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A101 of A172 Appendix to TNO-report of 3M-02 Final version 2:7-19 weight feces 7.3 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 393028 0.98 385167 87.2 87.9 12046 31.15 cholestanol 19805 0.91 18023 4.1 4.1 564 1.45 lathosterol 6961 1 6961 1.6 1.6 218 0.56 campesterol 5235 1.56 8167 1.8 1.9 255 0.64 stigmasterol 2749 1.49 4096 0.9 0.9 128 0.31 b-sitosterol 13255 1.47 19485 4.4 4.4 609 1.47 b-sitostanol 1 0 0.0 0.0 0 0.00 total 441033 441898 100 101 13820 35.6 cholestane 105292 1.04 2:7-20 weight feces 7 mg 109504 109504 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 363884 0.98 356606 86.1 91.7 13105 33.89 cholestanol 22163 0.91 20168 4.9 5.2 741 1.91 lathosterol 6943 1 6943 1.7 1.8 255 0.66 campesterol 5485 1.56 8557 2.1 2.2 314 0.78 stigmasterol 752 1.49 1120 0.3 0.3 41 0.10 b-sitosterol 14149 1.47 20799 5.0 5.4 764 1.84 b-sitostanol 1 0 0.0 0.0 0 0.00 total 413376 414194 100 107 15222 39.2 cholestane 93444 1.04 2:7-21 weight feces 6.9 mg 97182 97182 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 399924 0.98 391926 87.3 88.6 12845 33.22 cholestanol 19224 0.91 17494 3.9 4.0 573 1.48 lathosterol 7234 1 7234 1.6 1.6 237 0.61 campesterol 5395 1.56 8416 1.9 1.9 276 0.69 stigmasterol 2024 1.49 3016 0.7 0.7 99 0.24 b-sitosterol 14143 1.47 20790 4.6 4.7 681 1.64 b-sitostanol 1 0 0.0 0.0 0 0.00 total 447944 448876 100 102 14711 37.9 cholestane 106300 1.04 110552 110552 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A102 of A172 Appendix to TNO-report of 3M-02 Final version 3:7-19 weight feces 10 mg area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 582689 0.98 571035 88.5 114.6 11457 29.63 cholestanol 23901 0.91 21750 3.4 4.4 436 1.12 lathosterol 10390 1 10390 1.6 2.1 208 0.54 campesterol 7529 1.56 11745 1.8 2.4 236 0.59 stigmasterol 942 1.49 1404 0.2 0.3 28 0.07 b-sitosterol 19466 1.47 28615 4.4 5.7 574 1.38 b-sitostanol 1 0 0.0 0.0 0 0.00 total 644917 644939 100 129 12940 33.3 cholestane 119811 1.04 3:7-20 weight feces 4.9 mg 124603 124603 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 282329 0.98 276682 87.2 56.9 11609 30.02 cholestanol 16221 0.91 14761 4.7 3.0 619 1.59 lathosterol 5485 1 5485 1.7 1.1 230 0.60 campesterol 3642 1.56 5682 1.8 1.2 238 0.59 stigmasterol 437 1.49 651 0.2 0.1 27 0.07 b-sitosterol 9568 1.47 14065 4.4 2.9 590 1.42 b-sitostanol 1 0 0.0 0.0 0 0.00 total 317682 317326 100 65 13314 34.3 cholestane 116924 1.04 3:7-21 weight feces 9.5 mg 121601 121601 area response area corrected % g sterol ng sterol/ nmol sterol/ factor for response factor mg feces mg feces coprostanol 1 0 0.0 0.0 0 0.00 coprostanon 1 0 0.0 0.0 0 0.00 cholesterol 486322 0.98 476596 88.6 110.7 11653 30.13 cholestanol 21163 0.91 19258 3.6 4.5 471 1.21 lathosterol 7823 1 7823 1.5 1.8 191 0.49 campesterol 6293 1.56 9817 1.8 2.3 240 0.60 stigmasterol 622 1.49 927 0.2 0.2 23 0.05 b-sitosterol 16081 1.47 23639 4.4 5.5 578 1.39 b-sitostanol 1 0 0.0 0.0 0 0.00 total 538304 538060 100 125 13155 33.9 cholestane 103492 1.04 107632 107632 coprostanol coprostanon cholesterol cholestanol lathosterol campesterol stigmasterol b-sitosterol b-sitostanol 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 388.7 386.7 386.7 388.7 386.7 400.7 412.7 414.7 416.7 04-12-2006 Page A103 of A172 Page A104 of A172 group cage# GC neutral sterols (mol/g feces) neutral sterols total feces average weight days sample# coprostanol coprostanon cholesterol cholestanol lathosterol total (mol/g feces) (g) (g) 1 1,2 1:1-1 0.00 0.00 25.13 0.75 0.91 26.79 3.075 123.6 2 1 3,4 1:1-2 0.00 0.00 30.52 0.84 0.89 32.26 2.4897 87.25 2 1 5 1:1-3 0.00 0.00 26.76 0.71 0.94 28.40 2.2946 88.55 2 1 1,2 2:1-1 0.00 0.00 32.41 1.07 1.08 34.56 4.692 122.45 3 1 3,4 2:1-2 0.00 0.00 32.56 0.89 0.85 34.30 3.7789 86.55 3 1 5 2:1-3 0.00 0.00 34.17 0.93 1.14 36.25 3.4106 88.2 3 1 1,2 3:1-1 0.00 0.00 31.34 0.88 0.86 33.08 3.5754 121.65 2 1 3,4 3:1-2 0.00 0.00 27.10 0.78 0.75 28.64 2.8781 86.55 2 1 5 3:1-3 0.00 0.00 28.03 0.71 0.80 29.54 2.7589 88.05 2 AVERAGE SD 2 6 1:2-4 0.00 0.00 35.12 0.67 0.97 36.75 2.7619 94.55 2 2 7,8 1:2-5 0.00 0.00 40.98 0.52 1.33 42.83 4.0581 126.6 2 2 9,10 1:2-6 0.00 0.00 27.16 0.97 0.79 28.92 2.0935 82.2 2 2 6 2:2-4 0.00 0.00 35.56 0.80 0.98 37.33 4.075 94.25 3 2 7,8 2:2-5 0.00 0.00 39.75 0.75 1.24 41.74 6.371 126.45 3 2 9,10 2:2-6 0.00 0.00 31.14 1.09 0.80 33.04 3.4074 82.1 3 2 6 3:2-4 0.00 0.00 30.76 0.75 0.82 32.34 2.9279 94 2 2 7,8 3:2-5 0.00 0.00 29.95 0.55 0.88 31.38 4.3985 126.6 2 2 9,10 3:2-6 0.00 0.00 28.98 0.99 0.70 30.66 2.4575 81.85 2 AVERAGE SD 3 11,12,13 1:3-7 0.00 0.00 29.04 0.81 1.14 30.99 3.4597 118 2 3 15 1:3-8 0.00 0.00 26.49 0.95 0.79 28.23 2.2994 83.55 2 3 14,16,17 1:3-9 0.00 0.00 36.73 0.90 0.83 38.46 2.3454 82.8 2 3 11,12,13 2:3-7 0.00 0.00 30.71 0.80 1.11 32.62 5.8147 118.75 3 3 15 2:3-8 0.00 0.00 31.86 0.89 0.94 33.70 3.7718 83.5 3 3 14,16,17 2:3-9 0.00 0.00 36.75 0.94 0.83 38.52 3.7727 83.05 3 3 11,12,13 3:3-7 0.00 0.00 29.99 0.79 0.96 31.74 4.025 119.6 2 3 15 3:3-8 0.00 0.00 25.41 0.86 0.65 26.93 2.5265 84.2 2 3 14,16,17 3:3-9 0.00 0.00 30.71 0.83 0.71 32.25 2.6905 83.45 2 AVERAGE SD 4 18 1:4-10 0.00 0.00 35.76 0.86 0.80 37.41 1.689 59.5 2 4 19 1:4-11 0.00 0.00 34.60 0.98 0.78 36.37 3.1432 116.4 2 4 20,21 1:4-12 0.00 0.00 41.01 1.24 1.02 43.27 3.2545 112.2 2 4 18 2:4-10 0.00 0.00 47.77 1.26 1.03 50.07 2.7133 58.85 3 4 19 2:4-11 0.00 0.00 48.01 1.19 1.00 50.20 4.6407 116.55 3 4 20,21 2:4-12 0.00 0.00 49.87 1.21 1.07 52.15 5.2075 111.85 3 4 18 3:4-10 0.00 0.00 34.08 0.90 0.74 35.72 2.041 58.7 2 4 19 3:4-11 0.00 0.00 46.66 0.97 1.01 48.63 3.4464 117.45 2 4 20,21 3:4-12 0.00 0.00 40.89 0.94 0.88 42.71 3.9549 112.9 2 AVERAGE SD 5 22 1:5-13 0.00 0.00 42.83 1.24 1.18 45.25 3.195 110.7 2 5 24,25 1:5-14 0.00 0.00 36.13 0.66 0.94 37.73 2.9408 88.35 2 5 23,26,27 1:5-15 0.00 0.00 27.59 1.32 0.70 29.60 2.6399 84.6 2 5 22 2:5-13 0.00 0.00 40.79 1.13 1.18 43.10 4.9712 111.6 3 5 24,25 2:5-14 0.00 0.00 38.01 0.73 1.06 39.80 4.4833 88.5 3 5 23,26,27 2:5-15 0.00 0.00 28.11 1.66 0.74 30.52 3.767 85.55 3 5 22 3:5-13 0.00 0.00 32.13 0.82 0.86 33.81 3.6113 112.85 2 5 24,25 3:5-14 0.00 0.00 31.70 0.55 0.77 33.01 3.3522 88.9 2 5 23,26,27 3:5-15 0.00 0.00 22.66 1.20 0.52 24.38 2.7034 85.5 2 AVERAGE SD 6 28,29 1:6-16 0.00 0.00 35.38 1.10 0.90 37.38 2.5036 87.1 2 6 31 1:6-17 0.00 0.00 31.05 1.28 0.84 33.17 2.7083 112.8 2 6 30,32 1:6-18 0.00 0.00 33.74 1.13 0.89 35.76 2.3554 85.4 2 6 28,29 2:6-16 0.00 0.00 35.44 1.11 0.83 37.38 3.9684 87.15 3 6 31 2:6-17 0.00 0.00 32.61 1.59 0.80 35.00 3.9678 112.45 3 6 30,32 2:6-18 0.00 0.00 32.46 1.10 0.90 34.46 3.5666 84.55 3 6 28,29 3:6-16 0.00 0.00 33.26 0.92 0.70 34.88 2.9076 87.65 2 6 31 3:6-17 0.00 0.00 27.59 1.24 0.63 29.46 3.2449 111.95 2 6 30,32 3:6-18 0.00 0.00 36.14 1.15 0.96 38.25 2.7643 84.65 2 AVERAGE SD 7 33,34,35 1:7-19 0.00 0.00 33.92 1.76 0.59 36.27 1.9697 81.35 2 7 36,37 1:7-20 0.00 0.00 28.55 1.63 0.55 30.73 2.0798 82.25 2 7 38,39 1:7-21 0.00 0.00 30.49 1.56 0.56 32.61 2.6705 101.05 2 7 33,34,35 2:7-19 0.00 0.00 31.15 1.45 0.56 33.16 3.4477 82.35 3 7 36,37 2:7-20 0.00 0.00 33.89 1.91 0.66 36.46 3.2646 82.75 3 7 38,39 2:7-21 0.00 0.00 33.22 1.48 0.61 35.30 4.3059 101.95 3 7 33,34,35 3:7-19 0.00 0.00 29.63 1.12 0.54 31.29 2.5117 83.5 2 7 36,37 3:7-20 0.00 0.00 30.02 1.59 0.60 32.21 2.4369 83.35 2 7 38,39 3:7-21 0.00 0.00 30.13 1.21 0.49 31.84 3.4107 103.2 2 AVERAGE SD total neutrol sterols excretion (mol/100 gr mouse/day) 33.33 46.02 36.80 44.14 49.92 46.73 48.62 47.62 46.28 44.38 5.60 53.67 68.64 36.83 53.80 70.10 45.70 50.36 54.51 46.03 53.29 10.66 45.43 38.85 54.48 53.24 50.74 58.33 53.40 40.40 51.99 49.65 6.64 53.10 49.10 62.75 76.95 66.63 80.94 62.10 71.35 74.80 66.41 10.76 65.30 62.80 46.18 64.00 67.20 44.79 54.10 62.25 38.54 56.13 10.56 53.73 39.82 49.32 56.74 41.17 48.46 57.86 42.69 62.46 50.25 8.01 43.91 38.86 43.10 46.28 47.94 49.70 47.06 47.08 52.61 46.28 3.98 coprostanol 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 % of neutral sterols coprostanon cholesterol cholestanol 0.00 93.79 2.81 0.00 94.62 2.60 0.00 94.21 2.49 0.00 93.77 3.10 0.00 94.93 2.59 0.00 94.27 2.57 0.00 94.74 2.67 0.00 94.64 2.74 0.00 94.87 2.41 0.00 94.43 2.67 0.00 0.44 0.20 0.00 95.56 1.81 0.00 95.68 1.21 0.00 93.92 3.37 0.00 95.25 2.14 0.00 95.25 1.79 0.00 94.27 3.30 0.00 95.13 2.33 0.00 95.44 1.75 0.00 94.50 3.22 0.00 95.00 2.32 0.00 0.62 0.79 0.00 93.70 2.61 0.00 93.85 3.35 0.00 95.50 2.34 0.00 94.14 2.47 0.00 94.56 2.64 0.00 95.40 2.43 0.00 94.51 2.48 0.00 94.39 3.19 0.00 95.23 2.57 0.00 94.59 2.68 0.00 0.66 0.35 0.00 95.58 2.29 0.00 95.14 2.70 0.00 94.78 2.87 0.00 95.41 2.52 0.00 95.64 2.37 0.00 95.63 2.32 0.00 95.41 2.52 0.00 95.94 1.99 0.00 95.75 2.20 0.00 95.48 2.42 0.00 0.35 0.27 0.00 94.65 2.75 0.00 95.75 1.76 0.00 93.21 4.45 0.00 94.64 2.62 0.00 95.50 1.83 0.00 92.11 5.45 0.00 95.04 2.43 0.00 96.01 1.66 0.00 92.97 4.91 0.00 94.43 3.09 0.00 1.36 1.45 0.00 94.65 2.95 0.00 93.61 3.85 0.00 94.34 3.16 0.00 94.80 2.98 0.00 93.18 4.54 0.00 94.20 3.20 0.00 95.34 2.65 0.00 93.68 4.19 0.00 94.48 2.99 0.00 94.25 3.39 0.00 0.67 0.65 0.00 93.52 4.86 0.00 92.91 5.31 0.00 93.50 4.79 0.00 93.93 4.37 0.00 92.96 5.23 0.00 94.09 4.18 0.00 94.69 3.59 0.00 93.21 4.95 0.00 94.64 3.80 0.00 93.71 4.56 0.00 0.67 0.61 lathosterol 3.40 2.77 3.30 3.13 2.48 3.16 2.59 2.62 2.72 2.91 0.34 2.63 3.10 2.72 2.62 2.96 2.43 2.54 2.81 2.28 2.68 0.26 3.69 2.80 2.16 3.40 2.80 2.17 3.02 2.42 2.20 2.74 0.56 2.13 2.16 2.36 2.07 1.99 2.05 2.06 2.08 2.05 2.10 0.11 2.60 2.49 2.35 2.74 2.67 2.44 2.53 2.33 2.13 2.48 0.19 2.40 2.54 2.50 2.22 2.28 2.61 2.01 2.13 2.52 2.36 0.21 1.63 1.78 1.71 1.70 1.81 1.74 1.72 1.85 1.55 1.72 0.09 04-12-2006 Final version Appendix to TNO-report of 3M-02 Page A105 of A172 group cage# GC neutral sterols (mol/g feces) neutral sterols total feces average weight days sample# campesterol stigmasterol b-sitosterol b-sitostanol total (mol/g feces) (g) (g) 1 1,2 1:1-1 0.63 0.21 1.70 0.00 2.54 3.075 123.6 2 1 3,4 1:1-2 0.72 0.25 1.84 0.00 2.81 2.4897 87.25 2 1 5 1:1-3 0.60 0.20 1.51 0.00 2.31 2.2946 88.55 2 1 1,2 2:1-1 0.78 0.18 1.93 0.00 2.90 4.692 122.45 3 1 3,4 2:1-2 0.73 0.25 1.79 0.00 2.77 3.7789 86.55 3 1 5 2:1-3 0.75 0.24 1.94 0.00 2.93 3.4106 88.2 3 1 1,2 3:1-1 0.68 0.21 1.64 0.00 2.54 3.5754 121.65 2 1 3,4 3:1-2 0.66 0.16 1.64 0.00 2.46 2.8781 86.55 2 1 5 3:1-3 0.58 0.20 1.37 0.00 2.15 2.7589 88.05 2 AVERAGE SD 2 6 1:2-4 0.69 0.25 1.80 0.00 2.74 2.7619 94.55 2 2 7,8 1:2-5 0.64 0.24 1.64 0.00 2.52 4.0581 126.6 2 2 9,10 1:2-6 0.73 0.25 1.92 0.00 2.91 2.0935 82.2 2 2 6 2:2-4 0.70 0.12 1.76 0.00 2.59 4.075 94.25 3 2 7,8 2:2-5 0.74 0.27 1.85 0.00 2.86 6.371 126.45 3 2 9,10 2:2-6 0.80 0.17 2.07 0.00 3.04 3.4074 82.1 3 2 6 3:2-4 0.70 0.23 1.69 0.00 2.62 2.9279 94 2 2 7,8 3:2-5 0.56 0.16 1.25 0.00 1.97 4.3985 126.6 2 2 9,10 3:2-6 0.72 0.25 1.81 0.00 2.78 2.4575 81.85 2 AVERAGE SD 3 11,12,13 1:3-7 0.71 0.26 1.82 0.00 2.78 3.4597 118 2 3 15 1:3-8 0.67 0.24 1.69 0.00 2.60 2.2994 83.55 2 3 14,16,17 1:3-9 0.71 0.13 1.82 0.00 2.65 2.3454 82.8 2 3 11,12,13 2:3-7 0.74 0.26 1.84 0.00 2.83 5.8147 118.75 3 3 15 2:3-8 0.66 0.20 1.63 0.00 2.49 3.7718 83.5 3 3 14,16,17 2:3-9 0.71 0.13 1.85 0.00 2.70 3.7727 83.05 3 3 11,12,13 3:3-7 0.70 0.24 1.65 0.00 2.59 4.025 119.6 2 3 15 3:3-8 0.64 0.20 1.52 0.00 2.36 2.5265 84.2 2 3 14,16,17 3:3-9 0.64 0.14 1.56 0.00 2.33 2.6905 83.45 2 AVERAGE SD 4 18 1:4-10 0.70 0.26 1.78 0.00 2.74 1.689 59.5 2 4 19 1:4-11 0.53 0.08 1.34 0.00 1.94 3.1432 116.4 2 4 20,21 1:4-12 0.66 0.09 1.63 0.00 2.38 3.2545 112.2 2 4 18 2:4-10 0.67 0.10 1.68 0.00 2.45 2.7133 58.85 3 4 19 2:4-11 0.71 0.10 1.68 0.00 2.49 4.6407 116.55 3 4 20,21 2:4-12 0.77 0.08 1.86 0.00 2.72 5.2075 111.85 3 4 18 3:4-10 0.54 0.07 1.25 0.00 1.86 2.041 58.7 2 4 19 3:4-11 0.66 0.08 1.53 0.00 2.27 3.4464 117.45 2 4 20,21 3:4-12 0.61 0.07 1.42 0.00 2.10 3.9549 112.9 2 AVERAGE SD 5 22 1:5-13 0.70 0.13 1.80 0.00 2.64 3.195 110.7 2 5 24,25 1:5-14 0.59 0.24 1.50 0.00 2.32 2.9408 88.35 2 5 23,26,27 1:5-15 0.64 0.11 1.76 0.00 2.51 2.6399 84.6 2 5 22 2:5-13 0.72 0.11 1.74 0.00 2.57 4.9712 111.6 3 5 24,25 2:5-14 0.61 0.10 1.49 0.00 2.19 4.4833 88.5 3 5 23,26,27 2:5-15 0.79 0.14 1.97 0.00 2.90 3.767 85.55 3 5 22 3:5-13 0.59 0.22 1.47 0.00 2.27 3.6113 112.85 2 5 24,25 3:5-14 0.51 0.20 1.25 0.00 1.96 3.3522 88.9 2 5 23,26,27 3:5-15 0.58 0.11 1.36 0.00 2.04 2.7034 85.5 2 AVERAGE SD 6 28,29 1:6-16 0.66 0.09 1.71 0.00 2.47 2.5036 87.1 2 6 31 1:6-17 0.68 0.09 1.71 0.00 2.48 2.7083 112.8 2 6 30,32 1:6-18 0.66 0.09 1.73 0.00 2.48 2.3554 85.4 2 6 28,29 2:6-16 0.60 0.10 1.55 0.00 2.25 3.9684 87.15 3 6 31 2:6-17 0.77 0.07 1.91 0.00 2.75 3.9678 112.45 3 6 30,32 2:6-18 0.62 0.09 1.59 0.00 2.29 3.5666 84.55 3 6 28,29 3:6-16 0.53 0.07 1.27 0.00 1.87 2.9076 87.65 2 6 31 3:6-17 0.59 0.07 1.36 0.00 2.02 3.2449 111.95 2 6 30,32 3:6-18 0.66 0.08 1.61 0.00 2.34 2.7643 84.65 2 AVERAGE SD 7 33,34,35 1:7-19 0.75 0.10 1.91 0.00 2.76 1.9697 81.35 2 7 36,37 1:7-20 0.68 0.09 1.72 0.00 2.49 2.0798 82.25 2 7 38,39 1:7-21 0.72 0.09 1.77 0.00 2.57 2.6705 101.05 2 7 33,34,35 2:7-19 0.64 0.31 1.47 0.00 2.42 3.4477 82.35 3 7 36,37 2:7-20 0.78 0.10 1.84 0.00 2.73 3.2646 82.75 3 7 38,39 2:7-21 0.69 0.24 1.64 0.00 2.57 4.3059 101.95 3 7 33,34,35 3:7-19 0.59 0.07 1.38 0.00 2.04 2.5117 83.5 2 7 36,37 3:7-20 0.59 0.07 1.42 0.00 2.08 2.4369 83.35 2 7 38,39 3:7-21 0.60 0.05 1.39 0.00 2.05 3.4107 103.2 2 AVERAGE SD total neutrol sterols excretion (mol/100 gr mouse/day) 3.16 4.01 2.99 3.70 4.04 3.78 3.73 4.09 3.37 3.65 0.39 4.00 4.03 3.70 3.73 4.80 4.21 4.08 3.42 4.18 4.02 0.39 4.07 3.58 3.76 4.62 3.75 4.08 4.36 3.53 3.76 3.95 0.37 3.89 2.62 3.45 3.77 3.31 4.21 3.24 3.33 3.68 3.50 0.46 3.81 3.86 3.92 3.82 3.70 4.25 3.64 3.69 3.23 3.77 0.27 3.54 2.98 3.43 3.42 3.23 3.22 3.10 2.93 3.83 3.30 0.29 3.35 3.14 3.40 3.37 3.59 3.62 3.07 3.05 3.38 3.33 0.21 campesterol 24.96 25.48 25.94 27.00 26.27 25.66 26.91 26.93 26.95 26.23 0.76 25.12 25.53 25.18 27.14 25.78 26.27 26.64 28.32 26.02 26.22 1.03 25.38 25.76 26.71 26.04 26.43 26.48 26.94 27.07 27.51 26.48 0.67 25.58 27.20 27.65 27.46 28.49 28.28 28.79 28.92 29.11 27.94 1.11 26.55 25.24 25.40 27.78 27.70 27.10 25.86 25.80 28.26 26.63 1.12 26.89 27.35 26.66 26.85 28.05 26.95 28.47 28.98 28.13 27.59 0.83 27.15 27.35 27.87 26.37 28.77 26.78 28.82 28.54 29.25 27.88 1.02 % of neutral sterols stigmasterol b-sitosterol 8.33 66.71 8.91 65.61 8.77 65.29 6.32 66.68 9.15 64.58 8.15 66.18 8.45 64.63 6.49 66.57 9.21 63.85 8.20 65.57 1.08 1.05 9.29 65.59 9.40 65.07 8.72 66.10 4.77 68.09 9.45 64.77 5.69 68.05 8.70 64.66 8.21 63.47 8.84 65.13 8.12 65.66 1.70 1.54 9.20 65.42 9.09 65.15 4.91 68.38 9.07 64.89 8.02 65.55 4.95 68.57 9.39 63.67 8.48 64.46 5.81 66.68 7.66 65.86 1.89 1.69 9.66 64.76 3.87 68.93 3.67 68.68 3.96 68.58 4.01 67.50 3.09 68.63 3.91 67.30 3.42 67.66 3.42 67.46 4.33 67.72 2.02 1.28 5.09 68.37 10.23 64.53 4.47 70.14 4.45 67.77 4.41 67.88 4.81 68.09 9.49 64.65 10.34 63.86 5.19 66.55 6.50 66.87 2.67 2.12 3.70 69.41 3.73 68.92 3.70 69.64 4.27 68.88 2.50 69.44 3.84 69.21 3.65 67.88 3.65 67.37 3.38 68.49 3.60 68.81 0.47 0.76 3.56 69.28 3.42 69.22 3.36 68.77 12.84 60.79 3.66 67.57 9.32 63.91 3.34 67.84 3.18 68.28 2.68 68.06 5.04 67.08 3.55 2.85 b-sitostanol 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 04-12-2006 Final version Appendix to TNO-report of 3M-02 Appendix to TNO-report of 3M-02 Final version 04-12-2006 sterols total neutral sterols coprostanol coprostanon cholesterol cholestanol lathosterol total phytosterols campesterol stigmasterol b-sitosterol b-sitostanol sterols total neutral sterols coprostanol coprostanon cholesterol cholestanol lathosterol total phytosterols campesterol stigmasterol b-sitosterol b-sitostanol sterols total neutral sterols coprostanol coprostanon cholesterol cholestanol lathosterol total phytosterols campesterol stigmasterol b-sitosterol b-sitostanol group 1 vs 2 nn p 9 9 0.050 9 9 1.000 9 9 1.000 9 9 0.040 9 9 0.258 9 9 0.190 9 9 0.113 9 9 0.863 9 9 0.666 9 9 0.931 9 9 1.000 group 2 vs 4 nn p 9 9 0.040 9 9 1.000 9 9 1.000 9 9 0.077 9 9 0.489 9 9 <0.001 9 9 0.024 9 9 0.006 9 9 0.004 9 9 0.024 9 9 1.000 group 3 vs 7 nn p 9 9 0.190 9 9 1.000 9 9 1.000 9 9 0.031 9 9 <0.001 9 9 <0.001 9 9 <0.001 9 9 0.008 9 9 0.040 9 9 0.190 9 9 1.000 group 1 vs 3 nn p 9 9 0.077 9 9 1.000 9 9 1.000 9 9 0.863 9 9 0.546 9 9 0.489 9 9 0.222 9 9 0.546 9 9 0.863 9 9 1.000 9 9 1.000 group 2 vs 5 nn p 9 9 0.666 9 9 1.000 9 9 1.000 9 9 0.605 9 9 0.340 9 9 0.113 9 9 0.136 9 9 0.489 9 9 0.387 9 9 0.387 9 9 1.000 group 4 vs 5 nn p 9 9 0.113 9 9 1.000 9 9 1.000 9 9 0.094 9 9 0.605 9 9 <0.001 9 9 0.190 9 9 0.031 9 9 0.002 9 9 0.387 9 9 1.000 Sterol excretion, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 nn p nn p nn p 9 9 <0.001 9 9 0.050 9 9 0.161 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 <0.001 9 9 0.546 9 9 0.489 9 9 0.050 9 9 1.000 9 9 0.002 9 9 <0.001 9 9 0.008 9 9 0.010 9 9 0.489 9 9 0.730 9 9 0.077 9 9 0.002 9 9 0.489 9 9 0.014 9 9 0.004 9 9 0.258 9 9 <0.001 9 9 0.001 9 9 0.190 9 9 <0.001 9 9 1.000 9 9 1.000 9 9 1.000 group 2 vs 6 group 2 vs 7 group 3 vs 4 nn p nn p nn p 9 9 0.666 9 9 0.113 9 9 0.006 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 0.040 9 9 0.003 9 9 0.003 9 9 0.050 9 9 <0.001 9 9 0.136 9 9 0.008 9 9 <0.001 9 9 <0.001 9 9 0.001 9 9 <0.001 9 9 0.063 9 9 0.008 9 9 0.003 9 9 0.006 9 9 <0.001 9 9 0.031 9 9 0.004 9 9 <0.001 9 9 0.113 9 9 0.024 9 9 1.000 9 9 1.000 9 9 1.000 group 4 vs 6 group 4 vs 7 group 5 vs 6 nn p nn p nn p 9 9 0.006 9 9 <0.001 9 9 0.190 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 1.000 9 9 <0.001 9 9 <0.001 9 9 0.489 9 9 <0.001 9 9 <0.001 9 9 0.258 9 9 0.011 9 9 <0.001 9 9 0.297 9 9 0.190 9 9 0.340 9 9 0.004 9 9 0.258 9 9 0.796 9 9 0.094 9 9 0.436 9 9 0.297 9 9 <0.001 9 9 0.040 9 9 0.730 9 9 0.024 9 9 1.000 9 9 1.000 9 9 1.000 group 1 vs 7 nn p 9 9 0.605 9 9 1.000 9 9 1.000 9 9 0.024 9 9 <0.001 9 9 <0.001 9 9 0.063 9 9 0.003 9 9 0.050 9 9 0.040 9 9 1.000 group 3 vs 5 nn p 9 9 0.161 9 9 1.000 9 9 1.000 9 9 0.730 9 9 0.931 9 9 0.387 9 9 0.546 9 9 0.730 9 9 0.436 9 9 0.546 9 9 1.000 group 5 vs 7 nn p 9 9 0.094 9 9 1.000 9 9 1.000 9 9 0.190 9 9 0.063 9 9 <0.001 9 9 0.001 9 9 0.031 9 9 0.024 9 9 0.605 9 9 1.000 group 2 vs 3 nn p 9 9 0.489 9 9 1.000 9 9 1.000 9 9 0.222 9 9 0.190 9 9 0.931 9 9 0.796 9 9 0.387 9 9 0.605 9 9 0.730 9 9 1.000 group 3 vs 6 nn p 9 9 0.863 9 9 1.000 9 9 1.000 9 9 0.340 9 9 0.008 9 9 0.190 9 9 0.001 9 9 0.014 9 9 <0.001 9 9 0.001 9 9 1.000 group 6 vs 7 nn p 9 9 0.297 9 9 1.000 9 9 1.000 9 9 0.113 9 9 0.004 9 9 <0.001 9 9 0.863 9 9 0.546 9 9 0.387 9 9 0.094 9 9 1.000 p<0.05 Page A106 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 E4. Fecal fatty acid measurements 1:1-1 weight feces 8.6 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 13257 256605 2203 231285 22933 1279 2731 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 2.5 13.9 1619 7.09 48.4 269.5 31340 122.23 0.4 2.3 269 1.06 43.6 242.9 28247 99.29 4.3 24.1 2801 9.91 0.2 1.3 156 0.56 0.5 2.9 334 1.20 total 530293 100 557.0 64766 241.33 C15:0 76166 1:1-2 weight feces 10.1 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 16324 404265 3337 411875 37807 2492 1091 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.9 14.8 1465 6.42 46.1 366.5 36288 141.53 0.4 3.0 300 1.18 47.0 373.4 36971 129.95 4.3 34.3 3394 12.01 0.3 2.3 224 0.80 0.1 1.0 98 0.35 total 877191 100 795.3 78739 292.24 C15:0 88241 1:1-3 weight feces 8.8 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 18959 482473 4337 520971 55946 3057 6151 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.7 14.2 1618 7.09 44.2 362.4 41183 160.62 0.4 3.3 370 1.46 47.7 391.3 44469 156.30 5.1 42.0 4775 16.90 0.3 2.3 261 0.93 0.6 4.6 525 1.89 total 1091894 100 820.2 93201 345.18 C15:0 106504 2:1-1 weight feces 7.6 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 8193 1.8 7.5 993 4.35 210520 45.4 193.9 25508 99.49 1596 0.3 1.5 193 0.76 214647 46.3 197.7 26008 91.42 24494 5.3 22.6 2968 10.51 1701 0.4 1.6 206 0.73 2635 0.6 2.4 319 1.15 total 463786 100 427.1 56196 208.40 C15:0 86874 2:1-2 weight feces 5.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 7613 1.8 6.7 1172 5.13 201403 46.8 176.8 31014 120.96 1437 0.3 1.3 221 0.87 196561 45.7 172.5 30268 106.39 19865 4.6 17.4 3059 10.83 1352 0.3 1.2 208 0.74 2147 0.5 1.9 331 1.19 total 430378 100 377.8 66274 246.11 C15:0 91143 2:1-3 weight feces 7.2 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 9801 1.4 9.1 1262 5.52 284195 41.3 263.4 36587 142.70 2725 0.4 2.5 351 1.38 348778 50.7 323.3 44901 157.83 35118 5.1 32.6 4521 16.00 2252 0.3 2.1 290 1.03 4534 0.7 4.2 584 2.10 total 687403 100 637.2 88496 326.56 C15:0 86307 molw C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 228.4 256.4 254.4 284.5 282.5 280.5 278.5 MYSTRIC ACID PALMITIC ACID PALMITOLEIC ACID STEARIC ACID OLEIC ACID LINOLEIC ACID LINOLENIC ACID Page A107 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 3:1-1 weight feces 7.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 5463 1.2 7.5 1023 4.48 156085 35.1 213.4 29233 114.01 1332 0.3 1.8 249 0.98 248855 56.0 340.2 46607 163.82 26880 6.0 36.8 5034 17.82 1566 0.4 2.1 293 1.05 4158 0.9 5.7 779 2.80 total 444339 100 607.5 83219 304.96 C15:0 58514 3:1-2 weight feces 5.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 7866 1.5 7.3 1287 5.63 223765 43.6 208.6 36598 142.74 1702 0.3 1.6 278 1.09 248550 48.4 231.7 40651 142.89 26968 5.3 25.1 4411 15.61 1646 0.3 1.5 269 0.96 2844 0.6 2.7 465 1.67 total 513341 100 478.6 83959 310.59 C15:0 85813 3:1-3 weight feces 8.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 11108 1.5 9.5 1150 5.03 319405 42.1 274.4 33063 128.95 2509 0.3 2.2 260 1.02 380522 50.1 326.9 39390 138.45 37736 5.0 32.4 3906 13.83 2443 0.3 2.1 253 0.90 5057 0.7 4.3 523 1.88 total 758780 100 651.9 78546 290.07 C15:0 93112 1:2-4 weight feces 13.4 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 27893 1.2 20.7 1545 6.76 881387 39.5 654.1 48810 190.37 8053 0.4 6.0 446 1.75 1181866 53.0 877.0 65450 230.05 109775 4.9 81.5 6079 21.52 5394 0.2 4.0 299 1.06 17094 0.8 12.7 947 3.40 total 2231462 100 1655.9 123575 454.92 C15:0 107806 1:2-5 weight feces 9.2 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 31737 1.4 19.1 2081 9.11 923618 40.8 557.2 60560 236.19 93 0.0 0.1 6 0.02 1135022 50.2 684.7 74421 261.59 147397 6.5 88.9 9665 34.21 7383 0.3 4.5 484 1.73 16098 0.7 9.7 1056 3.79 total 2261348 100 1364.1 148272 546.64 C15:0 132620 1:2-6 weight feces 9 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 10087 1.7 6.6 728 3.19 262021 45.2 170.1 18905 73.73 2302 0.4 1.5 166 0.65 272616 47.0 177.0 19669 69.14 27573 4.8 17.9 1989 7.04 1825 0.3 1.2 132 0.47 3059 0.5 2.0 221 0.79 total 579483 100 376.3 41810 155.01 C15:0 123199 Page A108 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 2:2-4 weight feces 9.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 18615 1.6 15.3 1579 6.91 488224 42.0 401.8 41421 161.55 4357 0.4 3.6 370 1.45 581814 50.0 478.8 49361 173.50 60179 5.2 49.5 5106 18.07 3098 0.3 2.5 263 0.94 7073 0.6 5.8 600 2.15 total 1163360 100 957.4 98699 364.58 C15:0 97212 2:2-5 weight feces 9.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 18128 1.3 14.7 1575 6.90 549910 39.1 444.5 47793 186.40 5378 0.4 4.3 467 1.84 733331 52.1 592.7 63734 224.02 85796 6.1 69.3 7457 26.39 4289 0.3 3.5 373 1.33 10396 0.7 8.4 904 3.24 total 1407228 100 1137.4 122302 450.12 C15:0 98978 2:2-6 weight feces 5.9 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 5981 2.7 4.9 824 3.61 103637 47.0 84.2 14272 55.66 790 0.4 0.6 109 0.43 97059 44.0 78.9 13366 46.98 10984 5.0 8.9 1513 5.35 884 0.4 0.7 122 0.43 1019 0.5 0.8 140 0.50 total 220354 100 179.0 30345 112.97 C15:0 98463 3:2-4 weight feces 8.6 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 18311 1.7 15.1 1754 7.68 420511 39.9 346.4 40284 157.11 3649 0.3 3.0 350 1.37 545499 51.7 449.4 52257 183.68 56501 5.4 46.5 5413 19.16 3001 0.3 2.5 287 1.02 7025 0.7 5.8 673 2.42 total 1054497 100 868.7 101017 372.45 C15:0 97105 3:2-5 weight feces 7.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 14837 1.3 11.7 1515 6.63 453886 39.8 356.9 46352 180.78 4567 0.4 3.6 466 1.83 591094 51.9 464.8 60364 212.18 63874 5.6 50.2 6523 23.09 3296 0.3 2.6 337 1.20 7991 0.7 6.3 816 2.93 total 1139545 100 896.1 116374 428.65 C15:0 101736 3:2-6 weight feces 8.4 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 5585 1.4 4.9 585 2.56 169177 42.5 148.8 17709 69.07 1364 0.3 1.2 143 0.56 196952 49.5 173.2 20617 72.47 20806 5.2 18.3 2178 7.71 1530 0.4 1.3 160 0.57 2302 0.6 2.0 241 0.87 total 397716 100 349.7 41633 153.80 C15:0 90981 Page A109 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 1:3-7 weight feces 11 mg 2:3-7 weight feces 8.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 14517 354931 3055 369554 36484 2417 4404 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.8 10.1 914 4.00 45.2 245.8 22349 87.16 0.4 2.1 192 0.76 47.1 256.0 23269 81.79 4.6 25.3 2297 8.13 0.3 1.7 152 0.54 0.6 3.1 277 1.00 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 7895 202071 1616 212839 21090 1476 2397 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.8 6.9 833 3.65 45.0 177.0 21327 83.18 0.4 1.4 171 0.67 47.4 186.4 22464 78.96 4.7 18.5 2226 7.88 0.3 1.3 156 0.56 0.5 2.1 253 0.91 total 785362 100 544.0 49451 183.38 total 449384 100 393.7 47429 175.80 C15:0 115502 C15:0 91324 1:3-8 weight feces 11.1 mg 2:3-8 weight feces 7.6 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 20655 613614 5769 774403 72889 4402 10243 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.4 11.8 1062 4.65 40.9 350.2 31548 123.04 0.4 3.3 297 1.17 51.6 441.9 39815 139.95 4.9 41.6 3747 13.27 0.3 2.5 226 0.81 0.7 5.8 527 1.89 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 13707 374445 3351 433392 46483 2770 5202 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.6 9.9 1300 5.69 42.6 269.9 35519 138.53 0.4 2.4 318 1.25 49.3 312.4 41111 144.50 5.3 33.5 4409 15.61 0.3 2.0 263 0.94 0.6 3.8 493 1.77 total 1501975 100 857.2 77221 284.77 total 879350 100 633.9 83414 308.29 C15:0 140182 C15:0 110969 1:3-9 weight feces 13.4 mg 2:3-9 weight feces 7.4 mg area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces C14:0 26223 1.2 14.5 1081 4.73 C16:0 851207 38.6 470.3 35098 136.89 C16:1 7949 0.4 4.4 328 1.29 C18:0 1186581 53.8 655.6 48927 171.97 C18:1 108707 4.9 60.1 4482 15.87 C18:2 7434 0.3 4.1 307 1.09 C18:3 16281 0.7 9.0 671 2.41 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 9500 267057 2386 302932 29071 2020 3014 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.5 7.5 1014 4.44 43.4 210.9 28499 111.15 0.4 1.9 255 1.00 49.2 239.2 32327 113.63 4.7 23.0 3102 10.98 0.3 1.6 216 0.77 0.5 2.4 322 1.15 total 2204382 100 1218.0 90894 334.26 total 615980 100 486.4 65734 243.12 C15:0 144789 C15:0 101306 Page A110 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 3:3-7 weight feces 8.5 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 7543 193432 1421 205498 21652 1375 2581 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.7 6.4 753 3.30 44.6 164.2 19312 75.32 0.3 1.2 142 0.56 47.4 174.4 20517 72.12 5.0 18.4 2162 7.65 0.3 1.2 137 0.49 0.6 2.2 258 0.93 total 433502 100 367.9 43281 160.36 C15:0 94269 3:3-8 weight feces 6.5 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 6822 183086 1710 197892 21000 1164 2282 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.6 6.4 987 4.32 44.2 172.2 26486 103.30 0.4 1.6 247 0.97 47.8 186.1 28628 100.63 5.1 19.7 3038 10.75 0.3 1.1 168 0.60 0.6 2.1 330 1.19 total 413956 100 389.3 59885 221.76 C15:0 85077 3:3-9 weight feces 14 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 13974 420919 4038 511096 49923 3410 6144 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.4 12.4 883 3.87 41.7 372.5 26609 103.78 0.4 3.6 255 1.00 50.6 452.3 32310 113.57 4.9 44.2 3156 11.17 0.3 3.0 216 0.77 0.6 5.4 388 1.39 total 1009504 100 893.5 63818 235.56 C15:0 90391 1:4-10 weight feces 8.2 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 16048 1.4 11.1 1350 5.91 495415 41.9 341.8 41686 162.58 3727 0.3 2.6 314 1.23 599026 50.7 413.3 50404 177.17 56734 4.8 39.1 4774 16.90 3322 0.3 2.3 280 1.00 7505 0.6 5.2 631 2.27 total 1181777 100 815.4 99439 367.06 C15:0 115946 1:4-11 weight feces 8.8 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 13693 1.5 8.5 966 4.23 392497 42.8 243.6 27681 107.96 3202 0.3 2.0 226 0.89 454998 49.7 282.4 32089 112.79 43482 4.7 27.0 3067 10.86 2667 0.3 1.7 188 0.67 5468 0.6 3.4 386 1.38 total 916007 100 568.5 64603 238.78 C15:0 128901 1:4-12 weight feces 9.8 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 11857 1.3 12.4 1261 5.52 361984 40.3 377.2 38489 150.11 2570 0.3 2.7 273 1.07 460388 51.3 479.7 48952 172.06 51833 5.8 54.0 5511 19.51 3265 0.4 3.4 347 1.24 5863 0.7 6.1 623 2.24 total 897760 100 935.5 95457 351.76 C15:0 76774 Page A111 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 2:4-10 weight feces 7.1 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 10522 1.2 8.5 1196 5.23 344618 40.8 278.0 39156 152.71 2591 0.3 2.1 294 1.16 438053 51.9 353.4 49772 174.94 40213 4.8 32.4 4569 16.17 2602 0.3 2.1 296 1.05 5292 0.6 4.3 601 2.16 total 843891 100 680.8 95883 353.44 C15:0 99169 2:4-11 weight feces 8.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 13021 1.5 10.2 1170 5.12 381006 42.6 297.9 34238 133.53 3180 0.4 2.5 286 1.12 444595 49.7 347.6 39952 140.43 44296 5.0 34.6 3981 14.09 2954 0.3 2.3 265 0.95 4928 0.6 3.9 443 1.59 total 893980 100 698.9 80335 296.84 C15:0 102328 2:4-12 weight feces 7.2 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 12246 1.6 9.2 1273 5.58 332981 44.3 249.3 34627 135.05 2528 0.3 1.9 263 1.03 357246 47.5 267.5 37150 130.58 40701 5.4 30.5 4232 14.98 2826 0.4 2.1 294 1.05 3646 0.5 2.7 379 1.36 total 752174 100 563.2 78218 289.63 C15:0 106848 3:4-10 weight feces 8.5 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 10172 1.3 9.5 1112 4.87 310431 40.9 288.5 33946 132.39 2449 0.3 2.3 268 1.05 394129 51.9 366.3 43098 151.49 34766 4.6 32.3 3802 13.46 2260 0.3 2.1 247 0.88 4580 0.6 4.3 501 1.80 total 758787 100 705.3 82973 305.94 C15:0 86070 3:4-11 weight feces 9.8 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 20169 1.9 17.5 1789 7.83 422311 39.3 367.1 37462 146.11 3653 0.3 3.2 324 1.27 565589 52.6 491.7 50171 176.35 52273 4.9 45.4 4637 16.41 2913 0.3 2.5 258 0.92 7488 0.7 6.5 664 2.39 total 1074396 100 934.0 95305 351.28 C15:0 92026 3:4-12 weight feces 10.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 15056 1.1 13.7 1283 5.62 501243 38.2 456.9 42703 166.55 4206 0.3 3.8 358 1.41 708410 54.0 645.8 60353 212.14 68397 5.2 62.3 5827 20.63 4333 0.3 3.9 369 1.32 9322 0.7 8.5 794 2.85 total 1310967 100 1195.1 111688 410.51 C15:0 87759 Page A112 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 1:5-13 weight feces 8.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 13526 393457 3030 473949 44962 3228 5563 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.4 9.5 1140 4.99 42.0 275.2 33162 129.34 0.3 2.1 255 1.00 50.5 331.6 39947 140.41 4.8 31.5 3790 13.41 0.3 2.3 272 0.97 0.6 3.9 469 1.68 total 937715 100 656.0 79035 291.81 C15:0 114357 1:5-14 weight feces 11 mg area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces C14:0 33483 1.3 19.7 1792 7.84 C16:0 1006010 39.7 592.1 53832 209.95 C16:1 9704 0.4 5.7 519 2.04 C18:0 1300166 51.3 765.3 69572 244.54 C18:1 160875 6.3 94.7 8608 30.47 C18:2 7669 0.3 4.5 410 1.46 C18:3 18973 0.7 11.2 1015 3.65 total 2536880 100 1493.2 135749 499.96 C15:0 135913 1:5-15 weight feces 8.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 11073 294462 2464 315545 28429 1608 3367 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.7 7.3 884 3.87 44.8 195.1 23500 91.65 0.4 1.6 197 0.77 48.0 209.0 25183 88.52 4.3 18.8 2269 8.03 0.2 1.1 128 0.46 0.5 2.2 269 0.96 total 656948 100 435.2 52429 194.27 C15:0 120773 2:5-13 weight feces 8.1 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 13428 386269 3253 437237 45693 2448 5034 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.5 10.8 1331 5.83 43.2 310.0 38276 149.28 0.4 2.6 322 1.27 48.9 350.9 43327 152.29 5.1 36.7 4528 16.03 0.3 2.0 243 0.86 0.6 4.0 499 1.79 total 893362 100 717.1 88525 327.35 C15:0 99670 2:5-14 weight feces 9.9 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 23143 686470 6696 830835 105595 4899 10267 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.4 17.1 1731 7.58 41.2 508.5 51359 200.31 0.4 5.0 501 1.97 49.8 615.4 62160 218.49 6.3 78.2 7900 27.97 0.3 3.6 367 1.31 0.6 7.6 768 2.76 total 1667905 100 1235.4 124787 460.38 C15:0 108008 2:5-15 weight feces 8.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 7030 128252 1016 106592 11903 760 1127 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 2.7 6.9 833 3.65 50.0 126.1 15190 59.24 0.4 1.0 120 0.47 41.5 104.8 12624 44.37 4.6 11.7 1410 4.99 0.3 0.7 90 0.32 0.4 1.1 133 0.48 total 256680 100 252.3 30401 113.53 C15:0 81381 Page A113 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 3:5-13 weight feces 6.8 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 26211 674622 5831 907134 95082 4891 12523 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.5 10.5 1538 6.73 39.1 269.2 39589 154.40 0.3 2.3 342 1.35 52.5 362.0 53233 187.11 5.5 37.9 5580 19.75 0.3 2.0 287 1.02 0.7 5.0 735 2.64 total 1726294 100 688.9 101304 373.01 C15:0 200479 3:5-14 weight feces 5.3 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 12309 339628 3302 368609 45053 2365 4243 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.6 10.3 1940 8.49 43.8 283.7 53533 208.79 0.4 2.8 520 2.05 47.5 307.9 58101 204.22 5.8 37.6 7101 25.14 0.3 2.0 373 1.33 0.5 3.5 669 2.40 total 775509 100 647.9 122237 452.42 C15:0 95763 3:5-15 weight feces 8.1 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 4597 119586 872 128120 13678 939 1419 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.7 4.7 584 2.56 44.4 123.1 15199 59.28 0.3 0.9 111 0.44 47.6 131.9 16283 57.23 5.1 14.1 1738 6.15 0.3 1.0 119 0.43 0.5 1.5 180 0.65 total 269211 100 277.1 34215 126.73 C15:0 77711 1:6-16 weight feces 11.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 20396 1.2 12.5 1070 4.69 664144 40.5 407.7 34849 135.92 5241 0.3 3.2 275 1.08 854148 52.1 524.4 44819 157.54 80539 4.9 49.4 4226 14.96 5034 0.3 3.1 264 0.94 11097 0.7 6.8 582 2.09 total 1640599 100 1007.2 86086 317.21 C15:0 130309 1:6-17 weight feces 8.9 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 12220 1.4 8.4 939 4.11 366294 42.7 250.4 28137 109.74 3077 0.4 2.1 236 0.93 427401 49.9 292.2 32831 115.40 40324 4.7 27.6 3098 10.96 2415 0.3 1.7 186 0.66 5166 0.6 3.5 397 1.42 total 856897 100 585.8 65824 243.23 C15:0 117016 1:6-18 weight feces 10.8 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 17671 1.4 10.4 962 4.21 535579 41.3 315.0 29168 113.76 4357 0.3 2.6 237 0.93 659139 50.8 387.7 35897 126.18 67882 5.2 39.9 3697 13.09 4052 0.3 2.4 221 0.79 8388 0.6 4.9 457 1.64 total 1297068 100 762.9 70639 260.59 C15:0 136015 Page A114 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 2:6-16 weight feces 12.1 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 16368 1.4 12.4 1028 4.50 492848 43.0 374.7 30966 120.77 3736 0.3 2.8 235 0.92 567463 49.5 431.4 35654 125.32 55948 4.9 42.5 3515 12.44 3689 0.3 2.8 232 0.83 6084 0.5 4.6 382 1.37 total 1146136 100 871.4 72013 266.16 C15:0 105228 2:6-17 weight feces 8.5 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 7619 2.2 7.2 846 3.70 165470 48.1 156.1 18369 71.64 1010 0.3 1.0 112 0.44 151890 44.1 143.3 16862 59.27 15484 4.5 14.6 1719 6.08 1040 0.3 1.0 115 0.41 1620 0.5 1.5 180 0.65 total 344133 100 324.7 38204 142.20 C15:0 84780 2:6-18 weight feces 10.2 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 13778 1.8 9.5 928 4.06 361552 46.5 248.5 24363 95.02 2752 0.4 1.9 185 0.73 355381 45.7 244.3 23948 84.17 38817 5.0 26.7 2616 9.26 2125 0.3 1.5 143 0.51 3865 0.5 2.7 260 0.94 total 778270 100 534.9 52444 194.69 C15:0 116392 3:6-16 weight feces 9.6 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 11600 1.2 10.6 1109 4.86 381872 39.5 350.5 36507 142.38 3071 0.3 2.8 294 1.15 510655 52.8 468.7 48818 171.59 50174 5.2 46.0 4797 16.98 2963 0.3 2.7 283 1.01 6403 0.7 5.9 612 2.20 total 966738 100 887.2 92420 340.17 C15:0 87169 3:6-17 weight feces 7.6 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 6657 1.8 6.5 849 3.72 174284 45.8 168.9 22222 86.67 1200 0.3 1.2 153 0.60 175484 46.1 170.0 22375 78.65 19588 5.2 19.0 2498 8.84 1351 0.4 1.3 172 0.61 1768 0.5 1.7 225 0.81 total 380332 100 368.6 48494 179.90 C15:0 82557 3:6-18 weight feces 8.1 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 8700 1.3 8.3 1022 4.47 277658 41.8 264.1 32606 127.17 1872 0.3 1.8 220 0.86 334779 50.4 318.4 39313 138.18 34667 5.2 33.0 4071 14.41 1792 0.3 1.7 210 0.75 4339 0.7 4.1 510 1.83 total 663807 100 631.4 77952 287.68 C15:0 84105 Page A115 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 1:7-19 weight feces 10.7 mg 2:7-19 weight feces 12.4 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 9334 205542 1575 189750 23168 1823 1777 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 2.2 6.0 557 2.44 47.5 131.2 12263 47.83 0.4 1.0 94 0.37 43.8 121.1 11321 39.79 5.4 14.8 1382 4.89 0.4 1.2 109 0.39 0.4 1.1 106 0.38 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 10569 262993 1907 260336 26561 1861 2435 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.9 8.5 689 3.02 46.4 212.5 17137 66.84 0.3 1.5 124 0.49 45.9 210.3 16963 59.63 4.7 21.5 1731 6.13 0.3 1.5 121 0.43 0.4 2.0 159 0.57 total 432969 100 276.4 25831 96.09 total 566662 100 457.9 36924 137.09 C15:0 125320 C15:0 99012 1:7-20 weight feces 7.2 mg 2:7-20 weight feces 6 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 5300 116241 798 106821 14034 1332 1059 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 2.2 3.9 545 2.39 47.3 86.1 11953 46.62 0.3 0.6 82 0.32 43.5 79.1 10984 38.61 5.7 10.4 1443 5.11 0.5 1.0 137 0.49 0.4 0.8 109 0.39 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 4357 90363 573 77962 9578 799 815 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 2.4 3.7 608 2.66 49.0 75.7 12617 49.21 0.3 0.5 80 0.31 42.3 65.3 10886 38.26 5.2 8.0 1337 4.73 0.4 0.7 112 0.40 0.4 0.7 114 0.41 total 245585 100 181.8 25254 93.92 total 184447 100 154.5 25754 95.99 C15:0 108053 C15:0 95490 1:7-21 weight feces 10 mg 2:7-21 weight feces 8.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 9522 270616 2338 313018 37006 3195 3428 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.5 6.2 622 2.72 42.3 176.8 17684 68.97 0.4 1.5 153 0.60 49.0 204.6 20455 71.90 5.8 24.2 2418 8.56 0.5 2.1 209 0.74 0.5 2.2 224 0.80 C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 7650 212450 1476 217370 22360 1545 2300 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.6 5.6 642 2.81 45.7 155.1 17829 69.54 0.3 1.1 124 0.49 46.7 158.7 18242 64.12 4.8 16.3 1876 6.64 0.3 1.1 130 0.46 0.5 1.7 193 0.69 total 639123 100 417.7 41766 154.30 total 465151 100 339.6 39036 144.75 C15:0 122421 C15:0 109572 Page A116 of A172 Appendix to TNO-report of 3M-02 Final version 3:7-19 weight feces 5.8 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 4977 125240 723 123021 11441 691 1184 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.9 4.8 822 3.60 46.9 120.0 20682 80.66 0.3 0.7 119 0.47 46.0 117.8 20316 71.41 4.3 11.0 1889 6.69 0.3 0.7 114 0.41 0.4 1.1 196 0.70 total 267277 100 256.0 44138 163.94 C15:0 83524 3:7-20 weight feces 7.7 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 5155 125067 811 121012 14616 1143 1266 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.9 4.2 548 2.40 46.5 102.5 13306 51.90 0.3 0.7 86 0.34 45.0 99.1 12875 45.25 5.4 12.0 1555 5.50 0.4 0.9 122 0.43 0.5 1.0 135 0.48 total 269070 100 220.4 28627 106.31 C15:0 97655 3:7-21 weight feces 17 mg C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 area 11944 374593 3142 460518 44662 3197 5430 % g fatty ng fatty acid nmol fatty acid/ acid mg feces mg feces 1.3 10.3 603 2.64 41.5 321.7 18926 73.82 0.3 2.7 159 0.62 51.0 395.5 23268 81.78 4.9 38.4 2257 7.99 0.4 2.7 162 0.58 0.6 4.7 274 0.99 total 903486 100 776.0 45648 168.41 C15:0 93140 04-12-2006 Page A117 of A172 Page A118 of A172 group 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 AVERAGE SD cage# 1,2 3,4 5 1,2 3,4 5 1,2 3,4 5 GC sample# 1:1-1 1:1-2 1:1-3 2:1-1 2:1-2 2:1-3 3:1-1 3:1-2 3:1-3 C14:0 7.09 6.42 7.09 4.35 5.13 5.52 4.48 5.63 5.03 fatty acids (mol/g feces) C16:0 C16:1 C18:0 C18:1 122.23 1.06 99.29 9.91 141.53 1.18 129.95 12.01 160.62 1.46 156.30 16.90 99.49 0.76 91.42 10.51 120.96 0.87 106.39 10.83 142.70 1.38 157.83 16.00 114.01 0.98 163.82 17.82 142.74 1.09 142.89 15.61 128.95 1.02 138.45 13.83 C18:2 0.56 0.80 0.93 0.73 0.74 1.03 1.05 0.96 0.90 C18:3 1.20 0.35 1.89 1.15 1.19 2.10 2.80 1.67 1.88 fatty acids total (mol/g feces) 241.33 292.24 345.18 208.40 246.11 326.56 304.96 310.59 290.07 6 1:2-4 6.76 190.37 1.75 230.05 21.52 1.06 3.40 7,8 1:2-5 9.11 236.19 0.02 261.59 34.21 1.73 3.79 9,10 1:2-6 3.19 73.73 0.65 69.14 7.04 0.47 0.79 6 2:2-4 6.91 161.55 1.45 173.50 18.07 0.94 2.15 7,8 2:2-5 6.90 186.40 1.84 224.02 26.39 1.33 3.24 9,10 2:2-6 3.61 55.66 0.43 46.98 5.35 0.43 0.50 6 3:2-4 7.68 157.11 1.37 183.68 19.16 1.02 2.42 7,8 3:2-5 6.63 180.78 1.83 212.18 23.09 1.20 2.93 9,10 3:2-6 2.56 69.07 0.56 72.47 7.71 0.57 0.87 454.92 546.64 155.01 364.58 450.12 112.97 372.45 428.65 153.80 11,12,13 15 14,16,17 11,12,13 15 14,16,17 11,12,13 15 14,16,17 1:3-7 1:3-8 1:3-9 2:3-7 2:3-8 2:3-9 3:3-7 3:3-8 3:3-9 4.00 87.16 0.76 81.79 8.13 0.54 1.00 4.65 123.04 1.17 139.95 13.27 0.81 1.89 4.73 136.89 1.29 171.97 15.87 1.09 2.41 3.65 83.18 0.67 78.96 7.88 0.56 0.91 5.69 138.53 1.25 144.50 15.61 0.94 1.77 4.44 111.15 1.00 113.63 10.98 0.77 1.15 3.30 75.32 0.56 72.12 7.65 0.49 0.93 4.32 103.30 0.97 100.63 10.75 0.60 1.19 5.52 150.11 1.07 172.06 19.51 1.24 2.24 183.38 284.77 334.26 175.80 308.29 243.12 160.36 221.76 351.76 18 19 20,21 18 19 20,21 18 19 20,21 1:4-10 5.91 162.58 1.23 177.17 16.90 1.00 2.27 1:4-11 4.23 107.96 0.89 112.79 10.86 0.67 1.38 1:4-12 5.52 150.11 1.07 172.06 19.51 1.24 2.24 2:4-10 5.23 152.71 1.16 174.94 16.17 1.05 2.16 2:4-11 5.12 133.53 1.12 140.43 14.09 0.95 1.59 2:4-12 5.58 135.05 1.03 130.58 14.98 1.05 1.36 3:4-10 4.87 132.39 1.05 151.49 13.46 0.88 1.80 3:4-11 7.83 146.11 1.27 176.35 16.41 0.92 2.39 3:4-12 5.62 166.55 1.41 212.14 20.63 1.32 2.85 367.06 238.78 351.76 353.44 296.84 289.63 305.94 351.28 410.51 22 1:5-13 4.99 129.34 1.00 140.41 13.41 0.97 1.68 24,25 1:5-14 7.84 209.95 2.04 244.54 30.47 1.46 3.65 23,26,27 1:5-15 3.87 91.65 0.77 88.52 8.03 0.46 0.96 22 2:5-13 5.83 149.28 1.27 152.29 16.03 0.86 1.79 24,25 2:5-14 7.58 200.31 1.97 218.49 27.97 1.31 2.76 23,26,27 2:5-15 3.65 59.24 0.47 44.37 4.99 0.32 0.48 22 3:5-13 6.73 154.40 1.35 187.11 19.75 1.02 2.64 24,25 3:5-14 8.49 208.79 2.05 204.22 25.14 1.33 2.40 23,26,27 3:5-15 2.56 59.28 0.44 57.23 6.15 0.43 0.65 291.81 499.96 194.27 327.35 460.38 113.53 373.01 452.42 126.73 28,29 31 30,32 28,29 31 30,32 28,29 31 30,32 1:6-16 4.69 135.92 1.08 157.54 14.96 0.94 2.09 1:6-17 4.11 109.74 0.93 115.40 10.96 0.66 1.42 1:6-18 4.21 113.76 0.93 126.18 13.09 0.79 1.64 2:6-16 4.50 120.77 0.92 125.32 12.44 0.83 1.37 2:6-17 3.70 71.64 0.44 59.27 6.08 0.41 0.65 2:6-18 4.06 95.02 0.73 84.17 9.26 0.51 0.94 3:6-16 4.86 142.38 1.15 171.59 16.98 1.01 2.20 3:6-17 3.72 86.67 0.60 78.65 8.84 0.61 0.81 3:6-18 4.47 127.17 0.86 138.18 14.41 0.75 1.83 317.21 243.23 260.59 266.16 142.20 194.69 340.17 179.90 287.68 33,34,35 1:7-19 2.44 47.83 0.37 39.79 4.89 0.39 0.38 36,37 1:7-20 2.39 46.62 0.32 38.61 5.11 0.49 0.39 38,39 1:7-21 2.72 68.97 0.60 71.90 8.56 0.74 0.80 33,34,35 2:7-19 3.02 66.84 0.49 59.63 6.13 0.43 0.57 36,37 2:7-20 2.66 49.21 0.31 38.26 4.73 0.40 0.41 38,39 2:7-21 2.81 69.54 0.49 64.12 6.64 0.46 0.69 33,34,35 3:7-19 3.60 80.66 0.47 71.41 6.69 0.41 0.70 36,37 3:7-20 2.40 51.90 0.34 45.25 5.50 0.43 0.48 38,39 3:7-21 2.64 73.82 0.62 81.78 7.99 0.58 0.99 96.09 93.92 154.30 137.09 95.99 144.75 163.94 106.31 168.41 total feces average weight days (g) (g) 3.075 123.6 2 2.4897 87.25 2 2.2946 88.55 2 4.692 122.45 3 3.7789 86.55 3 3.4106 88.2 3 3.5754 121.65 2 2.8781 86.55 2 2.7589 88.05 2 2.7619 94.55 2 4.0581 126.6 2 2.0935 82.2 2 4.075 94.25 3 6.371 126.45 3 3.4074 82.1 3 2.9279 94 2 4.3985 126.6 2 2.4575 81.85 2 3.4597 118 2 2.2994 83.55 2 2.3454 82.8 2 5.8147 118.75 3 3.7718 83.5 3 3.7727 83.05 3 4.025 119.6 2 2.5265 84.2 2 2.6905 83.45 2 1.689 59.5 2 3.1432 116.4 2 3.2545 112.2 2 2.7133 58.85 3 4.6407 116.55 3 5.2075 111.85 3 2.041 58.7 2 3.4464 117.45 2 3.9549 112.9 2 3.195 110.7 2 2.9408 88.35 2 2.6399 84.6 2 4.9712 111.6 3 4.4833 88.5 3 3.767 85.55 3 3.6113 112.85 2 3.3522 88.9 2 2.7034 85.5 2 2.5036 87.1 2 2.7083 112.8 2 2.3554 85.4 2 3.9684 87.15 3 3.9678 112.45 3 3.5666 84.55 3 2.9076 87.65 2 3.2449 111.95 2 2.7643 84.65 2 1.9697 81.35 2 2.0798 82.25 2 2.6705 101.05 2 3.4477 82.35 3 3.2646 82.75 3 4.3059 101.95 3 2.5117 83.5 2 2.4369 83.35 2 3.4107 103.2 2 total fatty acid excretion (mol/100 gr mouse/day) 300.20 416.95 447.24 266.18 358.19 420.92 448.15 516.42 454.44 403.19 80.09 664.43 876.12 197.40 525.43 755.96 156.29 580.05 744.63 230.89 525.69 268.58 268.83 391.86 473.41 286.94 464.20 368.14 269.83 332.70 567.05 380.33 103.97 520.97 322.39 510.16 543.18 393.97 449.48 531.88 515.39 719.00 500.71 110.09 421.11 832.08 303.10 486.06 777.41 166.63 596.83 852.97 200.35 515.17 265.81 455.90 292.00 359.37 403.99 167.25 273.76 564.22 260.72 469.72 360.77 124.47 116.33 118.75 203.89 191.32 126.23 203.79 246.56 155.41 278.30 182.29 57.68 C14:0 2.94 2.20 2.05 2.09 2.09 1.69 1.47 1.81 1.74 2.01 0.42 1.49 1.67 2.06 1.90 1.53 3.19 2.06 1.55 1.66 1.90 0.53 2.18 1.63 1.42 2.08 1.85 1.83 2.06 1.95 1.57 1.84 0.26 1.61 1.77 1.57 1.48 1.73 1.93 1.59 2.23 1.37 1.70 0.26 1.71 1.57 1.99 1.78 1.65 3.21 1.81 1.88 2.02 1.96 0.49 1.48 1.69 1.62 1.69 2.60 2.09 1.43 2.07 1.55 1.80 0.38 2.54 2.54 1.77 2.20 2.77 1.94 2.20 2.26 1.57 2.20 0.39 C16:0 50.65 48.43 46.53 47.74 49.15 43.70 37.39 45.96 44.46 46.00 3.91 41.85 43.21 47.57 44.31 41.41 49.27 42.18 42.17 44.91 44.10 2.74 47.53 43.21 40.95 47.31 44.93 45.72 46.97 46.58 42.68 45.10 2.33 44.29 45.21 42.68 43.21 44.99 46.63 43.27 41.59 40.57 43.60 1.89 44.32 41.99 47.18 45.60 43.51 52.18 41.39 46.15 46.77 45.46 3.24 42.85 45.12 43.65 45.38 50.38 48.81 41.86 48.18 44.20 45.60 2.90 49.77 49.63 44.70 48.75 51.26 48.04 49.20 48.81 43.83 48.22 2.42 % of fatty acids C16:1 C18:0 C18:1 0.44 41.14 4.11 0.40 44.47 4.11 0.42 45.28 4.90 0.36 43.87 5.04 0.35 43.23 4.40 0.42 48.33 4.90 0.32 53.72 5.84 0.35 46.00 5.03 0.35 47.73 4.77 0.38 45.97 4.79 0.04 3.65 0.54 0.39 50.57 4.73 0.00 47.85 6.26 0.42 44.60 4.54 0.40 47.59 4.96 0.41 49.77 5.86 0.38 41.59 4.74 0.37 49.32 5.14 0.43 49.50 5.39 0.36 47.12 5.01 0.35 47.54 5.18 0.13 2.86 0.57 0.41 44.60 4.43 0.41 49.14 4.66 0.39 51.45 4.75 0.38 44.91 4.48 0.41 46.87 5.06 0.41 46.74 4.52 0.35 44.97 4.77 0.44 45.38 4.85 0.31 48.92 5.55 0.39 47.00 4.79 0.04 2.37 0.35 0.34 48.27 4.60 0.37 47.24 4.55 0.31 48.92 5.55 0.33 49.50 4.58 0.38 47.31 4.75 0.36 45.09 5.17 0.34 49.52 4.40 0.36 50.20 4.67 0.34 51.68 5.02 0.35 48.63 4.81 0.02 1.93 0.37 0.34 48.12 4.60 0.41 48.91 6.09 0.40 45.56 4.13 0.39 46.52 4.90 0.43 47.46 6.07 0.42 39.09 4.40 0.36 50.16 5.30 0.45 45.14 5.56 0.34 45.16 4.86 0.39 46.24 5.10 0.04 3.19 0.70 0.34 49.66 4.72 0.38 47.44 4.51 0.36 48.42 5.02 0.35 47.08 4.68 0.31 41.68 4.28 0.37 43.23 4.76 0.34 50.44 4.99 0.33 43.72 4.91 0.30 48.03 5.01 0.34 46.64 4.76 0.03 3.05 0.25 0.38 41.41 5.09 0.34 41.11 5.44 0.39 46.60 5.55 0.36 43.49 4.47 0.33 39.86 4.93 0.34 44.30 4.59 0.29 43.56 4.08 0.32 42.57 5.18 0.37 48.56 4.74 0.35 43.49 4.90 0.03 2.74 0.48 C18:2 0.23 0.27 0.27 0.35 0.30 0.32 0.34 0.31 0.31 0.30 0.04 0.23 0.32 0.30 0.26 0.30 0.38 0.28 0.28 0.37 0.30 0.05 0.30 0.28 0.33 0.32 0.30 0.32 0.31 0.27 0.35 0.31 0.02 0.27 0.28 0.35 0.30 0.32 0.36 0.29 0.26 0.32 0.31 0.03 0.33 0.29 0.24 0.26 0.28 0.28 0.27 0.29 0.34 0.29 0.03 0.30 0.27 0.30 0.31 0.29 0.26 0.30 0.34 0.26 0.29 0.03 0.40 0.52 0.48 0.32 0.41 0.32 0.25 0.41 0.34 0.38 0.09 C18:3 0.50 0.12 0.55 0.55 0.48 0.64 0.92 0.54 0.65 0.55 0.21 0.75 0.69 0.51 0.59 0.72 0.45 0.65 0.68 0.56 0.62 0.10 0.54 0.66 0.72 0.52 0.57 0.48 0.58 0.53 0.64 0.58 0.08 0.62 0.58 0.64 0.61 0.54 0.47 0.59 0.68 0.69 0.60 0.07 0.58 0.73 0.50 0.55 0.60 0.42 0.71 0.53 0.51 0.57 0.10 0.66 0.59 0.63 0.52 0.45 0.48 0.65 0.45 0.64 0.56 0.09 0.40 0.42 0.52 0.42 0.43 0.48 0.43 0.45 0.58 0.46 0.06 04-12-2006 Final version Appendix to TNO-report of 3M-02 Appendix to TNO-report of 3M-02 Final version 04-12-2006 fatty acid total fatty acids C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 fatty acid total fatty acids C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 fatty acid total fatty acids C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 group 1 vs 2 nn p 9 9 0.258 9 9 0.297 9 9 0.136 9 9 0.666 9 9 0.191 9 9 0.222 9 9 0.931 9 9 0.190 group 2 vs 4 nn p 9 9 0.489 9 9 0.546 9 9 1.000 9 9 0.019 9 9 0.489 9 9 0.190 9 9 0.863 9 9 0.546 group 3 vs 7 nn p 9 9 <0.001 9 9 0.040 9 9 0.011 9 9 0.019 9 9 0.006 9 9 0.546 9 9 0.031 9 9 0.002 group 1 vs 3 nn p 9 9 0.730 9 9 0.436 9 9 0.340 9 9 0.730 9 9 0.297 9 9 0.863 9 9 0.666 9 9 0.666 group 2 vs 5 nn p 9 9 1.000 9 9 0.605 9 9 0.436 9 9 0.730 9 9 0.340 9 9 0.730 9 9 0.489 9 9 0.297 group 4 vs 5 nn p 9 9 1.000 9 9 0.113 9 9 0.161 9 9 0.019 9 9 0.094 9 9 0.387 9 9 0.297 9 9 0.340 fatty acid excretion, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 nn p nn p nn p 9 9 0.040 9 9 0.387 9 9 0.546 9 9 0.094 9 9 0.436 9 9 0.190 9 9 0.050 9 9 0.436 9 9 0.489 9 9 0.136 9 9 0.605 9 9 0.077 9 9 0.050 9 9 0.546 9 9 0.546 9 9 0.931 9 9 0.387 9 9 0.863 9 9 0.863 9 9 0.387 9 9 0.387 9 9 0.387 9 9 0.863 9 9 1.000 group 2 vs 6 group 2 vs 7 group 3 vs 4 nn p nn p nn p 9 9 0.222 9 9 0.008 9 9 0.040 9 9 0.931 9 9 0.063 9 9 0.222 9 9 0.222 9 9 0.008 9 9 0.190 9 9 0.014 9 9 0.040 9 9 0.014 9 9 0.546 9 9 0.008 9 9 0.077 9 9 0.094 9 9 0.436 9 9 1.000 9 9 0.796 9 9 0.024 9 9 0.730 9 9 0.190 9 9 0.001 9 9 0.436 group 4 vs 6 group 4 vs 7 group 5 vs 6 nn p nn p nn p 9 9 0.040 9 9 <0.001 9 9 0.222 9 9 0.730 9 9 0.011 9 9 0.340 9 9 0.161 9 9 0.002 9 9 1.000 9 9 0.730 9 9 1.000 9 9 0.011 9 9 0.222 9 9 0.001 9 9 0.863 9 9 0.931 9 9 0.605 9 9 0.489 9 9 0.436 9 9 0.040 9 9 0.546 9 9 0.489 9 9 <0.001 9 9 1.000 group 1 vs 7 nn p 9 9 <0.001 9 9 0.222 9 9 0.136 9 9 0.136 9 9 0.136 9 9 0.605 9 9 0.019 9 9 0.031 group 3 vs 5 nn p 9 9 0.297 9 9 0.931 9 9 0.931 9 9 0.863 9 9 1.000 9 9 0.340 9 9 0.136 9 9 0.796 group 5 vs 7 nn p 9 9 0.004 9 9 0.136 9 9 0.050 9 9 0.019 9 9 0.063 9 9 0.605 9 9 0.011 9 9 0.014 group 2 vs 3 nn p 9 9 0.297 9 9 0.730 9 9 0.387 9 9 0.730 9 9 0.387 9 9 0.113 9 9 0.546 9 9 0.340 group 3 vs 6 nn p 9 9 0.730 9 9 0.666 9 9 0.796 9 9 0.008 9 9 1.000 9 9 0.863 9 9 0.161 9 9 0.666 group 6 vs 7 nn p 9 9 0.002 9 9 0.040 9 9 0.077 9 9 0.796 9 9 0.050 9 9 0.489 9 9 0.008 9 9 0.008 p<0.05 Page A119 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Fatty acid balance group 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 AVERAGE SD cage# 1,2 3,4 5 1,2 3,4 5 1,2 3,4 5 GC sample# 1:1-1 1:1-2 1:1-3 2:1-1 2:1-2 2:1-3 3:1-1 3:1-2 3:1-3 6 1:2-4 7,8 1:2-5 9,10 1:2-6 6 2:2-4 7,8 2:2-5 9,10 2:2-6 6 3:2-4 7,8 3:2-5 9,10 3:2-6 11,12,13 15 14,16,17 11,12,13 15 14,16,17 11,12,13 15 14,16,17 1:3-7 1:3-8 1:3-9 2:3-7 2:3-8 2:3-9 3:3-7 3:3-8 3:3-9 18 19 20,21 18 19 20,21 18 19 20,21 1:4-10 1:4-11 1:4-12 2:4-10 2:4-11 2:4-12 3:4-10 3:4-11 3:4-12 22 24,25 23,26,27 22 24,25 23,26,27 22 24,25 23,26,27 1:5-13 1:5-14 1:5-15 2:5-13 2:5-14 2:5-15 3:5-13 3:5-14 3:5-15 28,29 31 30,32 28,29 31 30,32 28,29 31 30,32 1:6-16 1:6-17 1:6-18 2:6-16 2:6-17 2:6-18 3:6-16 3:6-17 3:6-18 33,34,35 36,37 38,39 33,34,35 36,37 38,39 33,34,35 36,37 38,39 1:7-19 1:7-20 1:7-21 2:7-19 2:7-20 2:7-21 3:7-19 3:7-20 3:7-21 total fatty acids (mol/g feces) 241.33 292.24 345.18 208.40 246.11 326.56 304.96 310.59 290.07 454.92 546.64 155.01 364.58 450.12 112.97 372.45 428.65 153.80 183.38 284.77 334.26 175.80 308.29 243.12 160.36 221.76 351.76 367.06 238.78 351.76 353.44 296.84 289.63 305.94 351.28 410.51 291.81 499.96 194.27 327.35 460.38 113.53 373.01 452.42 126.73 317.21 243.23 260.59 266.16 142.20 194.69 340.17 179.90 287.68 96.09 93.92 154.30 137.09 95.99 144.75 163.94 106.31 168.41 total feces (g) 3.075 2.4897 2.2946 4.692 3.7789 3.4106 3.5754 2.8781 2.7589 2.7619 4.0581 2.0935 4.075 6.371 3.4074 2.9279 4.3985 2.4575 3.4597 2.2994 2.3454 5.8147 3.7718 3.7727 4.025 2.5265 2.6905 1.689 3.1432 3.2545 2.7133 4.6407 5.2075 2.041 3.4464 3.9549 3.195 2.9408 2.6399 4.9712 4.4833 3.767 3.6113 3.3522 2.7034 2.5036 2.7083 2.3554 3.9684 3.9678 3.5666 2.9076 3.2449 2.7643 1.9697 2.0798 2.6705 3.4477 3.2646 4.3059 2.5117 2.4369 3.4107 average weight days (g) 123.6 2 87.25 2 88.55 2 122.45 3 86.55 3 88.2 3 121.65 2 86.55 2 88.05 2 94.55 2 126.6 2 82.2 2 94.25 3 126.45 3 82.1 3 94 2 126.6 2 81.85 2 118 2 83.55 2 82.8 2 118.75 3 83.5 3 83.05 3 119.6 2 84.2 2 83.45 2 59.5 2 116.4 2 112.2 2 58.85 3 116.55 3 111.85 3 58.7 2 117.45 2 112.9 2 110.7 2 88.35 2 84.6 2 111.6 3 88.5 3 85.55 3 112.85 2 88.9 2 85.5 2 87.1 2 112.8 2 85.4 2 87.15 3 112.45 3 84.55 3 87.65 2 111.95 2 84.65 2 81.35 2 82.25 2 101.05 2 82.35 3 82.75 3 101.95 3 83.5 2 83.35 2 103.2 2 fatty acids excretion (mol/100 gr mouse/day) 300.20 416.95 447.24 266.18 358.19 420.92 448.15 516.42 454.44 403.19 80.09 664.43 876.12 197.40 525.43 755.96 156.29 580.05 744.63 230.89 525.69 268.58 268.83 391.86 473.41 286.94 464.20 368.14 269.83 332.70 567.05 380.33 103.97 520.97 322.39 510.16 543.18 393.97 449.48 531.88 515.39 719.00 500.71 110.09 421.11 832.08 303.10 486.06 777.41 166.63 596.83 852.97 200.35 515.17 265.81 455.90 292.00 359.37 403.99 167.25 273.76 564.22 260.72 469.72 360.77 124.47 116.33 118.75 203.89 191.32 126.23 203.79 246.56 155.41 278.30 182.29 57.68 food intake (g) 23.2 18 16.8 36.8 28.8 25.6 20.2 15.3 13.8 19.9 27.7 16.3 28.8 45.4 28.2 16.7 23.6 13.2 26.2 16.5 18.3 44.7 28.3 29.3 23.7 13.6 14.7 12 21.7 24 19.8 35.2 38.2 11.3 18.7 22.6 21.5 19.6 19.5 36.5 32.2 30.3 18.7 28 14.5 17.6 20.3 17.2 29.7 31 27.9 16.5 17.1 15 16.8 16.9 21.9 29.2 26.5 35 15.7 15 19.8 fatty acids uptake (mol/100 gr mouse/day) 5283.22 5806.79 5340.11 5639.32 6244.01 5446.40 4673.78 4975.69 4411.43 5924.08 6158.51 5581.42 5733.89 6737.13 6445.31 5000.55 5246.96 4539.25 6249.55 5558.62 6220.85 7063.37 6359.72 6620.12 5577.59 4546.28 4958.16 5676.67 5247.30 6020.71 6313.30 5667.20 6408.63 5418.38 4481.44 5634.35 5466.64 6244.23 6487.74 6137.15 6827.33 6646.00 4664.12 8865.14 4773.44 5687.53 5065.43 5668.91 6394.80 5172.97 6191.97 5298.60 4299.33 4987.62 5812.74 5783.36 6100.10 6653.61 6009.19 6441.97 5292.28 5065.41 5400.26 cholesterol balance (mol/100 gr mouse/day) 4983.02 5389.84 4892.87 5373.15 5885.82 5025.48 4225.64 4459.27 3956.98 4910.23 611.93 5259.65 5282.39 5384.03 5208.45 5981.17 6289.03 4420.50 4502.33 4308.36 5181.77 682.54 5980.71 5166.76 5747.45 6776.43 5895.52 6251.98 5307.76 4213.57 4391.11 5525.70 841.88 5155.70 4924.91 5510.55 5770.13 5273.22 5959.15 4886.51 3966.05 4915.34 5151.28 587.51 5045.52 5412.15 6184.64 5651.09 6049.92 6479.38 4067.29 8012.16 4573.08 5719.47 1159.56 5231.63 4773.43 5309.54 5990.81 5005.72 5918.21 4734.38 4038.62 4517.90 5057.81 634.70 5696.42 5664.61 5896.21 6462.29 5882.96 6238.18 5045.71 4910.00 5121.96 5657.59 537.83 molw C14:0 C16:0 C16:1 C18:0 C18:1 C18:2 C18:3 average 228.4 256.4 254.4 284.5 282.5 280.5 278.5 266.46 MYSTRIC ACID PALMITIC ACID PALMITOLEIC ACID STEARIC ACID OLEIC ACID LINOLEIC ACID LINOLENIC ACID g/mol diet: 15 % fat= 0.563 mmol per g diet Page A120 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 fatty acid balance balance fatty acid balance balance fatty acid balance balance group 1 vs 2 nn p 9 9 0.436 group 2 vs 4 nn p 9 9 0.863 group 3 vs 7 nn p 9 9 1.000 group 1 vs 3 nn p 9 9 0.113 group 2 vs 5 nn p 9 9 0.258 group 4 vs 5 nn p 9 9 0.222 fatty acid balance, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 nn p nn p nn p 9 9 0.489 9 9 0.063 9 9 0.730 group 2 vs 6 group 2 vs 7 group 3 vs 4 nn p nn p nn p 9 9 0.796 9 9 0.258 9 9 0.222 group 4 vs 6 group 4 vs 7 group 5 vs 6 nn p nn p nn p 9 9 0.730 9 9 0.136 9 9 0.136 group 1 vs 7 nn p 9 9 0.019 group 3 vs 5 nn p 9 9 0.863 group 5 vs 7 nn p 9 9 0.931 group 2 vs 3 nn p 9 9 0.546 group 3 vs 6 nn p 9 9 0.297 group 6 vs 7 nn p 9 9 0.094 p<0.05 Page A121 of A172 Appendix to TNO-report of 3M-02 Final version Appendix F. Intestinal cholesterol absorption Cholesterol absorption Oral gavage mix: 14 C dpm average: 76083.3 76547.5 75205.8 74174.5 73763.5 75669.7 75241 3H dpm average: 92065.2 92196.7 90472.4 88575.4 92526.8 92548.9 91398 14C/3H dosing: 0.823 Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 14C dpm feces 7900.00 9070.58 8759.79 7943.46 9442.29 9027.49 9996.01 10914.90 11495.20 8402.11 8966.49 9560.47 10754.70 11858.50 12382.30 8918.47 9795.44 9570.76 11342.40 10247.10 11420.10 10160.50 11533.30 11375.70 9360.62 9242.62 9409.35 10257.10 11572.10 11790.60 7215.69 6854.01 8420.46 7620.85 7592.86 7701.77 10718.20 9765.43 11045.10 8973.56 10118.90 10186.80 11415.30 11521.70 9926.39 9713.21 10522.10 10357.80 10622.10 10309.40 9363.58 9289.29 8492.80 9169.78 7720.92 8438.85 8631.31 10252.90 9662.34 10589.00 3H dpm feces 17857.0 20224.1 19669.7 AVERAGE 17369.9 20044.0 19314.7 AVERAGE 16226.5 17696.9 19154.0 AVERAGE 15319.6 16793.3 16635.3 AVERAGE 19111.4 20818.2 21352.2 AVERAGE 19316.9 21221.6 20766.7 AVERAGE 19935.8 17921.2 19737.7 AVERAGE 22122.3 24951.5 24849.9 AVERAGE 18676.0 18523.6 18338.6 AVERAGE 18752.4 20953.2 21592.7 AVERAGE 13209.8 12708.1 14817.4 AVERAGE 13562.2 13608.7 13685.9 AVERAGE 22219.9 20159.2 22642.2 AVERAGE 15531.3 17347.9 17603.2 AVERAGE 21059.0 21372.1 19181.1 AVERAGE 17538.7 18995.2 18714.0 AVERAGE 17065.5 16759.2 15186.6 AVERAGE 18158.2 16878.2 18096.8 AVERAGE 15528.4 17238.0 17527.6 AVERAGE 18655.8 17086.2 19019.0 AVERAGE 14/3H feces 0.442 0.449 0.445 0.445 0.457 0.471 0.467 0.465 0.616 0.617 0.600 0.611 0.548 0.534 0.575 0.552 0.563 0.570 0.580 0.571 0.462 0.462 0.461 0.461 0.569 0.572 0.579 0.573 0.459 0.462 0.458 0.460 0.501 0.499 0.513 0.504 0.547 0.552 0.546 0.548 0.546 0.539 0.568 0.551 0.562 0.558 0.563 0.561 0.482 0.484 0.488 0.485 0.578 0.583 0.579 0.580 0.542 0.539 0.518 0.533 0.554 0.554 0.553 0.554 0.622 0.615 0.617 0.618 0.512 0.503 0.507 0.507 0.497 0.490 0.492 0.493 0.550 0.566 0.557 0.557 % cholesterol absorption 45.89 43.48 25.78 32.90 30.67 43.95 30.38 44.15 38.73 33.38 33.03 31.87 41.10 29.56 35.27 32.73 24.92 38.39 40.11 32.30 04-12-2006 Page A122 of A172 Appendix to TNO-report of 3M-02 Final version Mouse# 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 14C dpm feces 8632.76 7757.78 8732.12 9317.58 10349.10 10995.40 12726.80 11922.70 10824.40 11803.80 11509.60 10446.20 9824.33 10536.10 10819.00 9944.17 9896.95 9024.08 11012.10 12345.10 12611.60 10539.20 11997.90 11733.20 8467.17 8615.23 7791.76 10611.20 11669.60 11704.20 6797.41 8479.36 8042.25 9667.22 10332.00 10290.40 9274.67 9043.46 8359.95 8337.87 8978.72 9290.98 9260.73 8926.69 8216.57 7388.65 7801.01 8272.11 8775.47 8627.50 7935.44 8265.72 7672.71 7372.15 6512.96 7565.00 7444.81 8569.40 9540.76 9523.59 8330.02 9412.73 9427.18 9607.51 9889.25 9677.26 10990.90 9651.99 11648.90 9120.82 10887.20 11023.60 7559.63 8832.90 9369.62 3H dpm feces 18263.4 16543.7 18151.9 AVERAGE 18797.2 20750.2 21398.2 AVERAGE 21922.9 20859.7 19073.0 AVERAGE 20531.4 19930.8 17926.4 AVERAGE 17823.8 19638.0 19717.7 AVERAGE 17861.6 17830.0 16251.3 AVERAGE 20530.5 22826.4 23740.9 AVERAGE 19138.3 21840.1 21335.4 AVERAGE 19713.7 19837.0 18343.0 AVERAGE 16118.3 17565.2 17995.9 AVERAGE 15090.2 18363.5 17655.2 AVERAGE 18299.0 19525.1 19416.7 AVERAGE 18965.3 18555.8 17064.5 AVERAGE 16938.5 18142.5 18500.1 AVERAGE 21618.5 20655.8 19301.2 AVERAGE 17101.3 18082.3 19128.3 AVERAGE 18331.8 18170.1 16944.8 AVERAGE 17166.9 16099.9 15533.3 AVERAGE 13424.4 15510.5 15298.9 AVERAGE 17514.8 19514.6 19814.4 AVERAGE 16006.8 17737.5 17550.5 AVERAGE 19624.7 19941.0 19580.0 AVERAGE 20009.6 17422.8 20445.0 AVERAGE 17747.4 20878.9 21294.1 AVERAGE 14314.6 16604.8 17203.4 AVERAGE 14/3H feces 0.473 0.469 0.481 0.474 0.496 0.499 0.514 0.503 0.581 0.572 0.568 0.573 0.575 0.577 0.583 0.578 0.551 0.537 0.549 0.545 0.557 0.555 0.555 0.556 0.536 0.541 0.531 0.536 0.551 0.549 0.550 0.550 0.430 0.434 0.425 0.430 0.658 0.664 0.650 0.658 0.450 0.462 0.456 0.456 0.528 0.529 0.530 0.529 0.489 0.487 0.490 0.489 0.492 0.495 0.502 0.496 0.428 0.432 0.426 0.429 0.432 0.431 0.432 0.432 0.479 0.475 0.468 0.474 0.481 0.477 0.475 0.478 0.485 0.488 0.487 0.487 0.489 0.489 0.481 0.486 0.520 0.531 0.537 0.529 0.490 0.496 0.494 0.493 0.549 0.554 0.570 0.558 0.514 0.521 0.518 0.518 0.528 0.532 0.545 0.535 % cholesterol absorption 42.39 38.93 30.37 29.74 33.74 32.50 34.87 33.19 47.82 20.11 44.62 35.72 40.63 39.69 47.92 47.53 42.43 41.99 40.90 40.93 35.69 40.08 32.26 37.12 35.02 04-12-2006 Page A123 of A172 Appendix to TNO-report of 3M-02 Final version Mouse# 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 14C dpm feces 13292.00 12025.90 8154.44 7073.99 8303.55 8312.14 8105.89 9177.91 8404.42 9083.21 10025.30 10551.50 7998.17 9589.24 9485.67 8469.79 9318.33 9497.10 8613.92 8699.27 7646.35 10299.70 9010.87 10286.90 8794.09 9327.04 9893.72 12083.00 11744.30 10287.20 7269.37 8544.20 8283.26 7909.58 7041.53 8328.59 7120.23 8146.99 8204.53 8975.84 9015.56 7523.90 7587.01 8719.17 8811.04 7582.08 8218.83 8867.68 8808.83 8584.87 7447.43 7758.45 6896.60 7801.31 8952.17 10058.20 10221.10 8171.80 7457.41 8538.29 8381.63 7890.69 7086.95 9800.20 11238.90 11334.00 6081.05 6823.98 6967.67 7436.52 8308.62 8710.60 8595.56 8773.69 7650.67 3H dpm feces 23039.1 20891.1 14093.8 AVERAGE 11977.8 13879.6 13866.1 AVERAGE 14355.8 16142.2 14482.0 AVERAGE 16293.9 17848.5 18335.8 AVERAGE 17522.8 20188.8 20422.8 AVERAGE 18261.6 19807.5 20101.5 AVERAGE 19823.8 19531.5 17499.6 AVERAGE 20630.3 17880.3 20622.3 AVERAGE 18229.9 19419.7 20500.1 AVERAGE 22066.1 21734.5 19073.9 AVERAGE 16991.1 19484.0 19099.1 AVERAGE 18224.2 16492.5 19376.2 AVERAGE 17677.8 19926.7 19132.5 AVERAGE 20407.6 19983.1 17563.4 AVERAGE 17288.6 19191.4 19513.2 AVERAGE 18143.7 20142.1 21451.9 AVERAGE 21295.7 20799.9 18297.0 AVERAGE 17723.8 15907.7 18207.4 AVERAGE 21000.2 23550.7 23744.5 AVERAGE 18430.1 17029.8 19260.2 AVERAGE 20381.9 19487.6 17369.0 AVERAGE 23428.2 26990.4 26923.6 AVERAGE 14094.2 15200.4 16000.0 AVERAGE 18241.1 20465.5 21554.3 AVERAGE 21140.4 21390.2 18677.5 AVERAGE 14/3H feces 0.577 0.576 0.579 0.577 0.591 0.598 0.599 0.596 0.565 0.569 0.580 0.571 0.557 0.562 0.575 0.565 0.456 0.475 0.464 0.465 0.464 0.470 0.472 0.469 0.435 0.445 0.437 0.439 0.499 0.504 0.499 0.501 0.482 0.480 0.483 0.482 0.548 0.540 0.539 0.542 0.428 0.439 0.434 0.433 0.434 0.427 0.430 0.430 0.403 0.409 0.429 0.413 0.440 0.451 0.428 0.440 0.439 0.454 0.452 0.448 0.418 0.408 0.413 0.413 0.414 0.413 0.407 0.411 0.438 0.434 0.428 0.433 0.426 0.427 0.430 0.428 0.443 0.438 0.443 0.442 0.411 0.405 0.408 0.408 0.418 0.416 0.421 0.419 0.431 0.449 0.435 0.439 0.408 0.406 0.404 0.406 0.407 0.410 0.410 0.409 % cholesterol absorption 29.90 27.59 30.62 31.38 43.48 43.04 46.68 39.18 41.48 34.11 47.36 47.73 49.77 46.58 45.55 49.82 50.06 47.37 48.02 46.36 50.43 49.16 46.72 50.69 50.34 04-12-2006 Page A124 of A172 Appendix to TNO-report of 3M-02 Group# 1 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 7 AVERAGE SD Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 % cholesterol absorption 45.89 43.48 25.78 32.90 30.67 43.95 30.38 44.15 38.73 33.38 36.9 7.2 33.03 31.87 41.10 29.56 35.27 32.73 24.92 38.39 40.11 32.30 33.9 5.0 42.39 38.93 30.37 29.74 33.74 32.50 34.87 33.19 47.82 20.11 34.4 7.5 44.62 35.72 40.63 39.69 47.92 47.53 42.43 41.99 40.90 40.93 42.2 3.7 35.69 40.08 32.26 37.12 35.02 29.90 27.59 30.62 31.38 43.48 34.3 4.9 43.04 46.68 39.18 41.48 34.11 47.36 47.73 49.77 46.58 45.55 44.1 4.7 49.82 50.06 47.37 48.02 46.36 50.43 49.16 46.72 50.69 50.34 48.9 1.6 Final version 04-12-2006 Page A125 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 week 8/9 week 8/9 week 8/9 group 1 vs 2 nn p 10 10 0.315 group 2 vs 4 nn p 10 10 <0.001 group 3 vs 7 nn p 10 10 <0.001 Intestinal cholesterol absorption, p values Mann-Whitney U-test group 1 vs 3 group 1 vs 4 group 1 vs 5 group 1 vs 6 group 1 vs 7 nn p nn p nn p nn p nn p 10 10 0.529 10 10 0.143 10 10 0.353 10 10 0.015 10 10 <0.001 group 2 vs 5 group 2 vs 6 group 2 vs 7 group 3 vs 4 group 3 vs 5 nn p nn p nn p nn p nn p 10 10 0.912 10 10 <0.001 10 10 <0.001 10 10 0.009 10 10 1.000 group 4 vs 5 group 4 vs 6 group 4 vs 7 group 5 vs 6 group 5 vs 7 nn p nn p nn p nn p nn p 10 10 0.002 10 10 0.315 10 10 <0.001 10 10 0.001 10 10 <0.001 group 2 vs 3 nn p 10 10 0.796 group 3 vs 6 nn p 10 10 0.005 group 6 vs 7 nn p 10 10 0.003 p<0.05 Page A126 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Appendix G. Liver weight and perigonadal fat weight and lipid contents liver G1. Liver weight and perigonadal fat weight group 1 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 7 AVERAGE SD Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 eartag LR 01 12 00 20 00 02 11 00 02 12 bodyweight (g) 34.5 31.9 30.4 28.9 32.4 30.1 28.4 32.8 32 28 1 0 35.2 2 1 30.3 02 34 0 1 34.9 1 0 30.4 0 1 31.3 2 0 32.9 0 0 29.2 2 1 28.8 1 0 31.2 01 31 02 31 0 1 33.6 1 1 29.8 1 0 27.7 0 1 30.9 2 0 31.2 1 2 26.3 0 2 32.3 1 0 29.6 1 0 28.3 0 2 28.5 0 0 29.1 1 0 29.8 0 1 31.1 0 2 26.4 0 0 27.6 1 1 30.1 0 0 27.8 1 1 28.3 1 0 31.8 0 1 25.9 1 1 29.6 1 2 29.2 1 2 29.6 2 0 30.2 0 1 31.9 1 1 30.1 0 0 29.2 1 2 29.6 01 28 1 0 31.2 1 2 25.9 1 0 27.5 0 2 26.9 0 0 25.4 0 1 26.6 0 2 27.8 22 30 2 1 26.4 2 0 29.9 10 30 0 2 28.7 0 0 30.3 1 2 29.4 0 0 28.3 0 0 28.1 20 28 10 25 01 28 perigonadal fat weight (g) 1.06 0.72 0.68 0.39 0.81 0.57 0.63 0.94 0.89 0.48 0.72 0.21 1.16 0.53 0.92 0.78 0.55 0.43 0.50 0.74 0.87 0.92 0.74 0.23 0.77 0.74 0.59 0.37 0.42 0.64 0.75 0.44 1.05 0.33 0.61 0.23 0.24 0.26 0.43 0.42 0.45 0.39 0.23 0.30 0.34 0.32 0.34 0.08 0.82 0.54 0.65 0.39 0.51 0.57 0.44 0.52 0.86 0.47 0.58 0.16 0.23 0.34 0.14 0.19 0.25 0.30 0.41 0.37 0.31 0.21 0.28 0.09 0.34 0.36 0.33 0.38 0.30 0.30 0.35 0.32 0.29 0.30 0.33 0.03 liver weight (g) 2.19 2 1.66 1.41 1.68 1.7 1.61 1.77 1.68 1.49 1.72 0.23 2.77 1.17 1.88 2.53 1.68 1.23 1.35 1.84 1.53 1.98 1.80 0.53 2 2.23 2.27 1.68 1.56 2.06 1.85 1.51 2.42 2.14 1.97 0.31 3.46 3.68 3.69 3.93 4.23 1.94 3.62 3.9 4.03 3.39 3.59 0.63 1.9 1.71 2.48 1.69 2.25 2.46 1.47 1.88 1.82 2.14 1.98 0.34 3.9 4.87 4.1 4.17 4.13 3.48 3.13 4.11 4.74 4.33 4.10 0.52 4.29 4.33 3.88 4.28 4.15 3.85 3.91 3.83 3.33 3.92 3.98 0.30 liver weight as % of bw (%) 6.35 6.27 5.46 4.88 5.19 5.65 5.67 5.40 5.25 5.32 5.54 0.46 7.87 3.86 5.53 7.25 5.53 3.93 4.10 6.30 5.31 6.35 5.60 1.38 6.45 7.19 6.76 5.64 5.63 6.67 5.93 5.74 7.49 7.23 6.47 0.71 12.23 12.91 12.68 13.19 13.60 7.35 13.12 12.96 14.50 11.98 12.45 1.92 5.97 6.60 8.38 5.79 7.60 8.15 4.61 6.25 6.23 7.23 6.68 1.16 13.93 15.61 15.83 15.16 15.35 13.70 11.77 14.78 15.80 16.40 14.83 1.37 14.35 14.43 13.52 14.13 14.12 13.60 13.91 13.68 13.32 14.00 13.91 0.37 Page A127 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 tissue perigonadal fat liver liver as % of bodyweight tissue perigonadal fat liver liver as % of bodyweight tissue perigonadal fat liver liver as % of bodyweight Liverweight; perigonadal fat weight; liverweight as % of bodyweight, p values Mann-Whitney U-test group 1 vs 2 group 1 vs 3 group 1 vs 4 group 1 vs 5 group 1 vs 6 group 1 vs 7 group 2 vs 3 nn p nn p nn p nn p nn p nn p nn p 10 10 0.912 10 10 0.315 10 10 <0.001 10 10 0.123 10 10 <0.001 10 10 <0.001 10 10 0.190 10 10 0.971 10 10 0.075 10 10 <0.001 10 10 0.043 10 10 <0.001 10 10 <0.001 10 10 0.247 10 10 0.796 10 10 0.003 10 10 <0.001 10 10 0.015 10 10 <0.001 10 10 <0.001 10 10 0.089 group 2 vs 4 group 2 vs 5 group 2 vs 6 group 2 vs 7 group 3 vs 4 group 3 vs 5 group 3 vs 6 nn p nn p nn p nn p nn p nn p nn p 10 10 <0.001 10 10 0.123 10 10 <0.001 10 10 <0.001 10 10 0.003 10 10 0.971 10 10 <0.001 10 10 <0.001 10 10 0.315 10 10 <0.001 10 10 <0.001 10 10 <0.001 10 10 0.912 10 10 <0.001 10 10 <0.001 10 10 0.105 10 10 <0.001 10 10 <0.001 10 10 <0.001 10 10 0.529 10 10 <0.001 group 3 vs 7 group 4 vs 5 group 4 vs 6 group 4 vs 7 group 5 vs 6 group 5 vs 7 group 6 vs 7 nn p nn p nn p nn p nn p nn p nn p 10 10 <0.001 10 10 <0.001 10 10 0.105 10 10 0.796 10 10 <0.001 10 10 <0.001 10 10 0.190 10 10 <0.001 10 10 <0.001 10 10 0.043 10 10 0.105 10 10 <0.001 10 10 <0.001 10 10 0.481 10 10 <0.001 10 10 <0.001 10 10 0.003 10 10 0.003 10 10 <0.001 10 10 <0.001 10 10 0.029 p<0.05 Page A128 of A172 Appendix to TNO-report of 3M-02 Final version G2. Lipid contents liver Liverhomogenates; protein concentration mouse 1 13 21 31 45 51 61 2 14 22 32 46 52 62 3 17 23 33 41 53 63 4 11 24 34 42 54 64 6 12 25 35 43 55 65 5 15 26 36 44 56 66 7 16 27 37 47 57 67 8 18 28 38 48 58 68 9 19 29 39 49 59 69 10 20 40 60 30 50 70 measured prot conc (mg/ml) well 1 well 2 0.551 0.56 0.292 0.301 0.701 0.686 1.058 1.098 0.384 0.389 0.468 0.46 1.208 1.279 0.468 0.46 0.479 0.468 0.596 0.587 1.011 0.949 0.707 0.692 1.406 1.44 1.185 1.185 0.35 0.341 0.59 0.579 0.768 0.772 0.622 0.607 0.484 0.494 1.022 1.091 0.942 0.945 0.75 0.734 0.734 0.71 0.508 0.516 0.747 0.722 0.407 0.407 1.054 1.047 0.778 0.765 0.489 0.468 0.185 0.187 0.263 0.274 0.822 0.829 0.546 0.573 1.251 1.231 1.627 1.608 0.524 0.529 0.579 0.579 0.533 0.541 0.296 0.321 0.758 0.749 0.707 0.71 0.453 0.455 0.468 0.478 0.433 0.425 0.438 0.438 1.099 1.151 0.13 0.13 0.93 0.934 1.953 2.04 0.428 0.415 0.36 0.351 0.463 0.458 1.124 1.147 0.481 0.465 0.372 0.363 0.779 0.801 0.465 0.478 0.568 0.554 0.776 0.755 0.173 0.169 0.62 0.612 2.018 2.035 0.692 0.674 0.321 0.314 0.341 0.341 0.239 0.235 0.355 0.348 0.175 0.177 0.743 0.728 1.444 1.462 dilution 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 Protein conc (mg/ml) 11.11 5.93 13.87 21.56 7.73 9.28 24.87 9.28 9.47 11.83 19.60 13.99 28.46 23.70 6.91 11.69 15.40 12.29 9.78 21.13 18.87 14.84 14.44 10.24 14.69 8.14 21.01 15.43 9.57 3.72 5.37 16.51 11.19 24.82 32.35 10.53 11.58 10.74 6.17 15.07 14.17 9.08 9.46 8.58 8.76 22.50 2.60 18.64 39.93 8.43 7.11 9.21 22.71 9.46 7.35 15.80 9.43 11.22 15.31 3.42 12.32 40.53 13.66 6.35 6.82 4.74 7.03 3.52 14.71 29.06 04-12-2006 Page A129 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 HPTLC measurements, liver lipids: mouse# 1 13 21 31 45 51 61 2 14 22 32 46 52 62 3 17 23 33 Peak# 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 Name Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Width [mm] 4.57 8.13 3.94 4.19 4.57 6.22 4.06 5.08 4.57 7.87 3.94 4.06 3.81 5.46 4.19 4.32 4.06 6.99 4.19 4.83 4.57 7.62 4.32 4.7 4.19 7.75 4.06 4.32 4.57 6.73 4.06 4.45 4.32 7.11 4.06 4.32 4.06 7.49 3.17 3.81 4.45 5.71 3.81 4.19 4.45 8.64 3.17 4.57 4.83 8 4.32 4.57 4.45 6.6 3.94 4.83 4.95 4.83 4.06 3.94 4.45 6.6 3.68 4.7 4.06 6.86 3.81 5.08 4.57 6.22 4.19 4.57 Pos [mm] 6.73 21.84 27.3 35.31 6.22 19.94 26.42 34.8 5.97 19.68 26.16 34.42 6.35 19.43 26.42 34.8 6.35 20.32 26.42 34.67 6.48 20.07 26.29 34.42 6.48 20.07 26.16 34.16 6.99 20.32 26.54 34.54 6.86 20.83 26.16 33.66 6.86 21.97 26.29 33.66 7.62 23.24 27.56 34.16 6.48 22.86 27.56 35.31 6.48 20.83 26.54 34.54 6.22 19.56 25.65 34.16 5.97 18.41 25.4 33.78 6.22 20.07 26.16 34.29 6.35 21.34 26.67 34.54 6.86 22.23 27.3 34.92 od-Bkg 62604 287988 22996 105220 52524 197880 24262 125607 43766 251729 26977 68880 34466 123478 27048 67378 52422 260322 24841 141768 50585 265253 24749 127056 48224 248371 28494 130184 44510 192428 25345 79085 54357 258846 23946 89672 57023 299353 22427 84768 39787 115041 21069 81470 51326 318787 21609 111253 41327 242241 25212 94035 46397 221527 27238 133113 35829 104710 25858 42916 44899 206251 22142 78487 51469 216844 25800 74388 46317 162768 26102 89651 g lipid/ 200 g protein 5.44 50.09 2.00 9.15 4.33 32.62 2.00 10.35 3.24 37.32 2.00 5.11 2.55 18.26 2.00 4.98 4.22 41.92 2.00 11.41 4.09 42.87 2.00 10.27 3.38 34.87 2.00 9.14 3.51 30.37 2.00 6.24 4.54 43.24 2.00 7.49 5.09 53.39 2.00 7.56 3.78 21.84 2.00 7.73 4.75 59.01 2.00 10.30 3.28 38.43 2.00 7.46 3.41 32.53 2.00 9.77 2.77 16.20 2.00 3.32 4.06 37.26 2.00 7.09 3.99 33.62 2.00 5.77 3.55 24.94 2.00 Page A130 of A172 6.87 Appendix to TNO-report of 3M-02 Final version 04-12-2006 mouse# 41 53 63 4 11 24 34 42 54 64 6 12 25 35 43 55 65 5 Peak# 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 Name Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Width [mm] 5.08 5.59 4.06 4.06 4.19 6.86 3.94 3.94 4.45 7.49 3.3 4.45 4.32 5.33 4.32 3.56 4.19 8.51 3.56 5.33 3.94 5.97 4.45 4.95 4.19 6.48 4.06 5.33 4.45 6.48 3.94 5.08 4.06 6.6 3.94 5.59 4.06 6.99 4.32 6.1 4.7 5.97 3.56 4.06 3.94 5.97 4.32 5.59 3.81 5.46 3.94 4.45 4.32 6.73 4.06 4.95 4.7 7.24 3.94 4.95 3.68 6.6 3.56 5.59 3.81 6.6 3.81 5.84 3.56 5.71 4.19 5.59 Pos [mm] 7.37 22.61 27.69 35.31 7.24 22.99 27.81 35.31 6.73 23.11 27.43 35.05 7.37 23.37 28.45 36.07 5.46 21.21 26.16 34.04 5.46 18.8 25.27 33.66 4.95 17.02 24 33.02 4.57 16 23.24 32.64 4.7 15.75 23.11 32.64 5.21 16 23.24 32.64 6.22 19.68 26.29 34.29 4.45 15.37 22.86 32.13 5.33 17.14 24.13 33.02 5.59 19.05 24.77 33.15 5.46 20.57 25.53 33.4 3.94 13.97 21.21 30.35 3.94 13.84 20.96 30.35 4.19 13.08 20.7 30.23 od-Bkg 39765 132487 27644 37392 49635 246286 23327 123642 60183 294481 22239 141459 38024 104943 29874 26558 35962 307860 24049 107934 40358 115325 25364 52857 42991 172245 24996 73646 56968 192630 26017 123337 56224 215795 27127 153105 55767 238689 31249 162067 53651 199383 25618 75588 38266 151763 32037 54371 49570 149995 26617 46177 61124 219127 28657 145012 56257 295337 28462 97524 48925 249297 28191 157664 65477 276227 27578 178277 49411 189703 29682 95600 g lipid/ 200 g protein 2.88 19.17 2.00 2.71 4.26 42.23 2.00 10.60 5.41 52.97 2.00 12.72 2.55 14.05 2.00 1.78 2.99 51.20 2.00 8.98 3.18 18.19 2.00 4.17 3.44 27.56 2.00 5.89 4.38 29.62 2.00 9.48 4.15 31.82 2.00 11.29 3.57 30.55 2.00 10.37 4.19 31.13 2.00 5.90 2.39 18.95 2.00 3.39 3.72 22.54 2.00 3.47 4.27 30.59 2.00 10.12 3.95 41.51 2.00 6.85 3.47 35.37 2.00 11.19 4.75 40.07 2.00 12.93 3.33 25.57 2.00 6.44 Page A131 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 mouse# 15 26 36 44 56 66 7 16 27 37 47 57 67 8 18 28 38 48 Peak# 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 Name Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Width [mm] 3.3 5.71 4.19 5.33 3.43 7.49 3.81 5.59 3.94 6.35 3.81 5.46 3.56 5.71 3.81 4.95 3.94 6.6 3.68 5.33 4.19 7.37 3.68 5.46 4.45 5.97 4.19 4.7 4.7 6.35 4.19 3.68 4.83 6.99 4.7 4.57 4.32 6.22 4.06 4.45 3.94 6.6 3.81 4.06 4.19 5.71 3.68 4.06 4.7 7.49 3.43 5.08 3.81 5.59 3.81 4.06 3.94 5.84 3.94 4.45 4.32 5.71 3.81 4.45 4.06 5.59 3.81 4.45 4.32 6.6 3.17 4.7 Pos [mm] 3.94 12.57 20.45 30.1 3.94 13.84 21.08 30.73 4.32 14.48 22.1 31.5 4.19 14.73 22.48 31.62 4.57 16.13 23.37 32.26 5.33 18.16 24.51 33.02 5.33 18.92 25.27 33.27 5.59 20.32 25.91 33.27 5.84 21.21 27.69 35.69 5.71 20.45 26.92 34.92 5.97 20.07 26.42 34.54 5.97 19.3 25.65 33.78 6.48 20.45 25.65 33.66 6.99 20.45 25.91 33.66 7.11 21.21 25.91 33.53 6.48 20.83 25.53 32.89 7.37 22.23 26.8 33.91 7.75 23.88 27.43 34.16 od-Bkg 51117 186121 31056 108021 51578 314298 30069 134094 49352 209014 27111 117556 43993 173724 24940 61245 53573 223574 24814 161398 55464 270083 24336 153330 51342 176085 23750 76943 47717 148024 25114 60069 45004 193690 27970 60245 43432 150439 25551 98765 42568 194874 23418 72154 40720 177809 27101 69768 52458 292884 29963 160643 50307 162127 27192 70970 56774 225869 27244 91446 49756 171023 26319 87109 41083 144165 27516 61184 55902 285090 26432 119215 g lipid/ 200 g protein 3.29 23.97 2.00 6.96 3.43 41.81 2.00 8.92 3.64 30.84 2.00 8.67 3.53 27.86 2.00 4.91 4.32 36.04 2.00 13.01 4.56 44.39 2.00 12.60 4.32 29.66 2.00 6.48 3.80 23.58 2.00 4.78 3.22 27.70 2.00 4.31 3.40 23.55 2.00 7.73 3.64 33.29 2.00 6.16 3.01 26.24 2.00 5.15 3.50 39.10 2.00 10.72 3.70 23.85 2.00 5.22 4.17 33.16 2.00 6.71 3.78 25.99 2.00 6.62 2.99 20.96 2.00 4.45 4.23 43.14 2.00 9.02 Page A132 of A172 Appendix to TNO-report of 3M-02 Final version mouse# 58 68 9 19 29 39 49 59 69 10 20 30 40 50 60 70 Peak# 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 Name Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Free cholesterol Triglycerides Internal standard Cholesterolester Width [mm] 3.81 6.73 3.43 4.57 3.56 8.89 4.06 5.71 3.17 5.59 3.81 4.95 3.3 4.7 3.94 4.95 3.3 6.99 3.68 5.33 3.56 5.08 4.19 5.33 3.94 4.83 4.32 4.95 3.94 5.97 4.06 5.71 3.17 8.13 4.32 5.46 3.43 5.21 3.68 4.83 3.43 6.22 3.68 4.7 3.17 5.08 3.68 4.32 4.06 5.46 2.92 4.83 3.56 4.83 3.43 3.94 4.19 6.35 2.79 4.83 4.7 7.49 3.17 4.45 Pos [mm] 7.11 23.24 27.43 34.16 4.57 14.61 22.1 32.13 4.57 12.19 20.83 31.24 4.32 11.68 20.57 31.12 3.81 12.57 20.57 31.12 3.81 11.68 20.7 31.24 4.06 11.81 21.34 32.13 3.81 12.57 21.84 32.38 4.7 14.86 22.99 33.4 4.57 13.59 22.99 33.27 4.83 14.99 23.75 33.91 5.08 15.49 24.38 34.16 8.76 27.43 30.1 35.18 8.89 25.78 29.34 35.18 8 24.77 28.57 35.05 7.49 23.11 27.81 34.8 od-Bkg 50057 254148 23859 136982 59495 368098 29699 172395 47364 186870 29695 97139 44604 114977 29508 85494 55495 290613 29976 133502 54475 146150 30815 126727 63475 140542 33224 84388 64340 238585 30644 139703 56623 368599 30442 178078 37007 124055 26723 78711 47883 216520 26901 83366 34039 136553 27780 92480 54261 222500 24418 136191 36509 87808 28028 35295 58652 238639 21843 134627 58311 269150 31958 124736 g lipid/ 200 g protein 4.20 42.61 2.00 11.48 4.01 49.58 2.00 11.61 3.19 25.17 2.00 6.54 3.02 15.59 2.00 5.79 3.70 38.78 2.00 8.91 3.54 18.97 2.00 8.23 3.82 16.92 2.00 5.08 4.20 31.14 2.00 9.12 3.72 48.43 2.00 11.70 2.77 18.57 2.00 5.89 3.56 32.20 2.00 6.20 2.45 19.66 2.00 6.66 4.44 36.45 2.00 11.15 2.61 12.53 2.00 2.52 5.37 43.70 2.00 12.33 3.65 33.69 2.00 7.81 04-12-2006 Page A133 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group# 1 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 7 AVERAGE SD Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 Liver lipids (g/200 g protein) Free cholesterol Triglycerides Cholesterolester 5.44 50.09 9.15 3.51 30.37 6.24 2.77 16.20 3.32 2.55 14.05 1.78 3.33 25.57 6.44 4.19 31.13 5.90 4.32 29.66 6.48 3.70 23.85 5.22 3.19 25.17 6.54 2.77 18.57 5.89 2.99 51.20 8.98 2.39 18.95 3.39 4.33 32.62 10.35 4.54 43.24 7.49 3.29 23.97 6.96 3.80 23.58 4.78 4.06 37.26 7.09 4.17 33.16 6.71 3.02 15.59 5.79 3.56 32.20 6.20 3.24 37.32 5.11 5.09 53.39 7.56 3.99 33.62 5.77 3.18 18.19 4.17 3.72 22.54 3.47 3.43 41.81 8.92 3.22 27.70 4.31 3.78 25.99 6.62 3.70 38.78 8.91 2.61 12.53 2.52 2.55 18.26 4.98 3.78 21.84 7.73 3.55 24.94 6.87 3.44 27.56 5.89 4.27 30.59 10.12 3.64 30.84 8.67 3.40 23.55 7.73 2.99 20.96 4.45 3.54 18.97 8.23 2.45 19.66 6.66 2.88 19.17 2.71 4.38 29.62 9.48 3.95 41.51 6.85 3.53 27.86 4.91 4.22 41.92 11.41 4.75 59.01 10.30 3.64 33.29 6.16 4.23 43.14 9.02 3.82 16.92 5.08 5.37 43.70 12.33 4.09 42.87 10.27 3.28 38.43 7.46 4.26 42.23 10.60 4.15 31.82 11.29 3.47 35.37 11.19 4.32 36.04 13.01 3.01 26.24 5.15 4.20 42.61 11.48 4.20 31.14 9.12 4.44 36.45 11.15 3.38 34.87 9.14 3.41 32.53 9.77 5.41 52.97 12.72 3.57 30.55 10.37 4.75 40.07 12.93 4.56 44.39 12.60 3.50 39.10 10.72 4.01 49.58 11.61 3.72 48.43 11.70 3.65 33.69 7.81 Liver lipids (g/mg protein) Free cholesterol Triglycerides Cholesterolester 27.22 250.5 45.76 17.56 151.8 31.20 13.86 81.0 16.60 12.73 70.3 8.89 16.65 127.8 32.21 20.94 155.7 29.51 21.62 148.3 32.40 18.50 119.2 26.10 15.95 125.9 32.71 13.85 92.8 29.45 17.89 132.3 28.48 4.41 51.0 9.92 14.95 256.0 44.88 11.94 94.7 16.97 21.65 163.1 51.77 22.70 216.2 37.45 16.46 119.9 34.78 19.00 117.9 23.92 20.28 186.3 35.45 20.84 165.8 33.57 15.12 77.9 28.97 17.80 161.0 30.99 18.07 155.9 33.87 3.44 55.1 9.86 16.22 186.6 25.53 25.43 267.0 37.80 19.95 168.1 28.83 15.91 90.9 20.84 18.62 112.7 17.35 17.15 209.0 44.59 16.09 138.5 21.54 18.90 130.0 33.10 18.51 193.9 44.54 13.03 62.7 12.59 17.98 155.9 28.67 3.28 61.0 11.14 12.74 91.3 24.91 18.88 109.2 38.67 17.74 124.7 34.35 17.20 137.8 29.46 21.33 152.9 50.60 18.20 154.2 43.36 17.00 117.8 38.65 14.93 104.8 22.24 17.68 94.9 41.13 12.25 98.3 33.29 16.80 118.6 35.67 2.78 23.2 8.64 14.38 95.9 13.53 21.90 148.1 47.41 19.77 207.5 34.26 17.64 139.3 24.56 21.10 209.6 57.07 23.75 295.1 51.49 18.18 166.4 30.81 21.15 215.7 45.10 19.10 84.6 25.40 26.85 218.5 61.64 20.38 178.1 39.13 3.45 63.8 15.76 20.44 214.4 51.34 16.39 192.2 37.30 21.28 211.2 53.00 20.73 159.1 56.44 17.35 176.9 55.93 21.59 180.2 65.04 15.03 131.2 25.74 20.98 213.0 57.41 21.00 155.7 45.59 22.22 182.2 55.77 19.70 181.6 50.36 2.49 27.5 11.40 16.92 174.3 45.69 17.03 162.7 48.87 27.06 264.8 63.61 17.85 152.8 51.86 23.74 200.3 64.65 22.79 222.0 63.01 17.51 195.5 53.61 20.03 247.9 58.05 18.60 242.2 58.50 18.25 168.4 39.03 19.98 203.1 54.69 3.43 39.4 8.45 Page A134 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 lipid free cholesterol triglyceride cholesterolester lipid free cholesterol triglyceride cholesterolester lipid free cholesterol triglyceride cholesterolester group 1 vs 2 nn p 10 10 0.739 10 10 0.247 10 10 0.218 group 2 vs 4 nn p 10 10 0.353 10 10 0.089 10 10 0.684 group 3 vs 7 nn p 10 10 0.247 10 10 0.075 10 10 <0.001 group 1 vs 3 nn p 10 10 0.796 10 10 0.353 10 10 0.912 group 2 vs 5 nn p 10 10 0.218 10 10 0.436 10 10 0.529 group 4 vs 5 nn p 10 10 0.023 10 10 0.035 10 10 0.579 lipid contents liver, p values Mann-Whitney U-test group 1 vs 4 group 1 vs 5 group 1 vs 6 nn p nn p nn p 10 10 0.853 10 10 0.105 10 10 0.218 10 10 0.631 10 10 0.105 10 10 0.004 10 10 0.105 10 10 0.190 10 10 0.002 group 2 vs 6 group 2 vs 7 group 3 vs 4 nn p nn p nn p 10 10 0.315 10 10 0.353 10 10 0.529 10 10 0.280 10 10 0.052 10 10 0.143 10 10 0.004 10 10 <0.001 10 10 0.143 group 4 vs 6 group 4 vs 7 group 5 vs 6 nn p nn p nn p 10 10 0.052 10 10 0.075 10 10 0.739 10 10 <0.001 10 10 <0.001 10 10 0.971 10 10 0.005 10 10 <0.001 10 10 0.105 group 1 vs 7 nn p 10 10 0.190 10 10 0.002 10 10 <0.001 group 3 vs 5 nn p 10 10 0.089 10 10 0.353 10 10 0.105 group 5 vs 7 nn p 10 10 0.579 10 10 0.315 10 10 0.015 group 2 vs 3 nn p 10 10 0.739 10 10 0.912 10 10 0.280 group 3 vs 6 nn p 10 10 0.089 10 10 0.247 10 10 0.001 group 6 vs 7 nn p 10 10 0.971 10 10 0.315 10 10 0.393 p<0.05 Page A135 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 H. HMG-CoA reductase and cholesterol-7-hydroxylase activity in liver microsomes and sterol27-hydroxylase activity in liver mitochondria H1. HMG-CoA reductase and cholesterol-7-hydroxylase activity in liver microsomes Protein concentration isolated microsomes Microsomes for HMG CoA reductase determination mouse 1 13 21 31 45 51 61 2 14 22 32 46 52 62 3 17 23 33 41 53 63 4 11 24 34 42 54 64 6 12 25 35 43 55 65 5 15 26 36 44 56 66 7 16 27 37 47 57 67 8 18 28 38 48 58 68 9 19 29 39 49 59 69 10 20 40 60 30 50 70 measured prot conc (mg/ml) well 1 well 2 0.828 0.866 0.725 0.739 1.052 1.102 1.42 1.525 0.927 0.977 0.659 0.654 0.654 0.688 0.763 0.801 1.019 1.073 0.788 0.818 1.667 1.803 1.031 1.056 0.64 0.647 0.691 0.709 0.789 0.828 0.669 0.688 0.718 0.739 0.876 0.888 0.664 0.677 1.86 1.983 1.733 1.851 0.583 0.604 0.752 0.773 0.718 0.729 1.124 1.161 0.734 0.752 1.555 1.696 1.756 1.961 0.7 0.723 0.519 0.521 0.822 0.846 1.223 1.308 1.136 1.176 1.649 1.791 0.651 0.643 0.622 0.628 0.98 1.041 0.98 0.991 0.763 0.796 0.881 0.91 1.149 1.216 1.2 1.281 0.66 0.734 0.597 0.64 1.127 1.198 1.254 1.315 0.67 0.699 1.697 1.712 1.641 1.823 0.637 0.668 0.794 0.826 0.832 0.859 1.63 1.553 0.796 0.817 1.747 1.899 0.636 0.632 1.097 1.149 0.736 0.766 0.92 0.906 1.503 1.47 0.98 1.034 0.621 0.612 1.154 1.219 0.798 0.82 0.807 0.794 1.17 1.203 1.685 1.612 0.629 0.657 1.092 1.048 1.529 1.598 dilution 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 60 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 60 20 20 20 20 20 60 20 20 20 20 20 20 20 20 Protein conc (mg/ml) 16.94 14.64 21.54 29.45 19.04 39.39 40.26 15.64 20.92 16.06 34.70 20.87 38.61 42.00 16.17 13.57 14.57 17.64 13.41 38.43 35.84 11.87 15.25 14.47 22.85 14.86 32.51 37.17 14.23 10.40 16.68 25.31 23.12 34.40 38.82 12.50 20.21 19.71 15.59 17.91 23.65 24.81 13.94 12.37 23.25 25.69 13.69 34.09 34.64 13.05 16.20 16.91 31.83 16.13 36.46 38.04 22.46 15.02 18.26 29.73 20.14 36.99 23.73 16.18 16.01 23.73 32.97 12.86 21.40 31.27 Microsomes for 7-alpha-hydroxylase determination mouse 1 13 21 31 45 51 61 2 14 22 32 46 52 62 3 17 23 33 41 53 63 4 11 24 34 42 54 64 6 12 25 35 43 55 65 5 15 26 36 44 56 66 7 16 27 37 47 57 67 8 18 28 38 48 58 68 9 19 29 39 49 59 69 10 20 40 60 30 50 70 Code 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 1.10 1.11 1.12 1.13 1.14 1.15 1.16 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.9 2.10 2.11 2.12 2.13 2.14 2.15 2.16 3.1 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 3.10 3.11 3.12 3.13 3.14 3.15 3.16 4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 4.10 4.11 4.12 4.13 4.14 4.15 4.16 5.1 5.2 5.3 5.4 5.5 5.6 measured prot conc (mg/ml) well 1 well 2 0.952 0.996 0.683 0.703 1.097 1.048 1.543 1.438 1.222 1.218 0.652 0.644 0.612 0.694 1.103 1.113 1.366 1.329 0.999 0.963 1.524 1.447 0.921 0.909 0.569 0.575 0.577 0.571 0.708 0.679 0.553 0.545 0.918 0.874 1.15 1.14 0.69 0.694 0.5 0.436 0.543 0.479 0.599 0.569 0.887 0.837 0.726 0.724 0.882 0.893 0.772 0.829 0.541 0.515 0.61 0.579 0.604 0.624 0.488 0.498 0.674 0.67 1.263 1.329 1.023 1.055 0.507 0.472 0.505 0.492 0.595 0.581 0.808 0.79 0.994 0.988 0.673 0.646 1.164 1.183 0.472 0.488 0.385 0.376 0.792 0.772 0.561 0.592 0.942 0.933 1.404 1.527 0.933 0.948 0.462 0.479 0.35 0.366 0.74 0.699 0.818 0.821 0.837 0.859 1.604 1.483 0.829 0.824 0.45 0.46 0.468 0.455 0.821 0.865 0.832 0.826 0.916 0.954 1.164 1.132 0.893 0.951 0.455 0.468 0.432 0.41 0.821 0.845 0.994 1.039 1.268 1.517 0.483 0.525 0.779 0.785 0.919 0.87 0.553 0.511 dilution 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 20 20 60 60 20 20 20 60 20 20 60 Protein conc (mg/ml) 19.48 13.86 21.45 29.81 24.40 38.88 39.18 22.16 26.95 19.62 29.71 18.30 34.32 34.44 13.87 10.98 17.92 22.90 13.84 28.08 30.66 11.68 17.24 14.50 17.75 16.01 31.68 35.67 12.28 9.86 13.44 25.92 20.78 29.37 29.91 11.76 15.98 19.82 13.19 23.47 28.80 22.83 15.64 11.53 18.75 29.31 18.81 28.23 21.48 14.39 16.39 16.96 30.87 16.53 27.30 27.69 16.86 16.58 18.70 22.96 18.44 27.69 25.26 16.66 20.33 27.85 30.24 15.64 17.89 31.92 Page A136 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Amount isolated liver microsomes group# mouse# 1 1 1 2 1 3 1 4 1 5 1 6 1 7 1 8 1 9 1 10 AVERAGE SD 2 11 2 12 2 13 2 14 2 15 2 16 2 17 2 18 2 19 2 20 AVERAGE SD 3 21 3 22 3 23 3 24 3 25 3 26 3 27 3 28 3 29 3 30 AVERAGE SD 4 31 4 32 4 33 4 34 4 35 4 36 4 37 4 38 4 39 4 40 AVERAGE SD 5 41 5 42 5 43 5 44 5 45 5 46 5 47 5 48 5 49 5 50 AVERAGE SD 6 51 6 52 6 53 6 54 6 55 6 56 6 57 6 58 6 59 6 60 AVERAGE SD 7 61 7 62 7 63 7 64 7 65 7 66 7 67 7 68 7 69 7 70 AVERAGE SD Protein conc (mg/ml) microsomes microsomes 7 alpha hydroxylase HMGCo A reduct. 19.48 16.94 22.16 15.64 13.87 16.17 11.68 11.87 11.76 12.50 12.28 14.23 15.64 13.94 14.39 13.05 16.86 22.46 16.66 16.18 estimated l microsomes 600 600 600 600 600 600 600 600 600 600 mg microsomes 7 alpha hydroxylase 11.688 13.296 8.322 7.008 7.056 7.368 9.384 8.634 10.116 9.996 mg microsomes HMGCo A reduct. 10.164 9.384 9.702 7.122 7.5 8.538 8.364 7.83 13.476 9.708 17.24 9.86 13.86 26.95 15.98 11.53 10.98 16.39 16.58 20.33 15.25 600 10.40 600 14.64 600 20.92 600 20.21 600 12.37 600 13.57 600 16.20 600 15.02 600 16.01 600 10.344 5.916 8.316 16.17 9.588 6.918 6.588 9.834 9.948 12.198 9.15 6.24 8.784 12.552 12.126 7.422 8.142 9.72 9.012 9.606 21.45 19.62 17.92 14.5 13.44 19.82 18.75 16.96 18.7 15.64 21.54 600 16.06 600 14.57 600 14.47 600 16.68 600 19.71 600 23.25 600 16.91 600 18.26 600 12.86 600 12.87 11.772 10.752 8.7 8.064 11.892 11.25 10.176 11.22 9.384 12.924 9.636 8.742 8.682 10.008 11.826 13.95 10.146 10.956 7.716 29.81 29.71 22.9 17.75 25.92 13.19 29.31 30.87 22.96 27.85 29.45 700 34.70 700 17.64 700 22.85 700 25.31 700 15.59 600 25.69 700 31.83 700 29.73 700 23.73 700 20.867 20.797 16.03 12.425 18.144 7.914 20.517 21.609 16.072 19.495 20.615 24.29 12.348 15.995 17.717 9.354 17.983 22.281 20.811 16.611 13.84 16.01 20.78 23.47 24.4 18.3 18.81 16.53 18.44 17.89 13.41 600 14.86 600 23.12 600 17.91 600 19.04 600 20.87 600 13.69 600 16.13 600 20.14 600 21.40 600 8.304 9.606 12.468 14.082 14.64 10.98 11.286 9.918 11.064 10.734 8.046 8.916 13.872 10.746 11.424 12.522 8.214 9.678 12.084 12.84 38.88 34.32 28.08 31.68 29.37 28.8 28.23 27.3 27.69 30.24 39.39 700 38.61 700 38.43 700 32.51 700 34.40 700 23.65 700 34.09 700 36.46 700 36.99 700 32.97 700 27.216 24.024 19.656 22.176 20.559 20.16 19.761 19.11 19.383 21.168 27.573 27.027 26.901 22.757 24.08 16.555 23.863 25.522 25.893 23.079 39.18 34.44 30.66 35.67 29.91 22.83 21.48 27.69 25.26 31.92 40.26 700 42.00 700 35.84 700 37.17 700 38.82 700 24.81 700 34.64 700 38.04 700 23.73 700 31.27 700 27.426 24.108 21.462 24.969 20.937 15.981 15.036 19.383 17.682 22.344 28.182 29.4 25.088 26.019 27.174 17.367 24.248 26.628 16.611 21.889 isolated microsomes (mg) 21.85 22.68 18.02 14.13 14.56 15.91 17.75 16.46 23.59 19.70 18.47 3.37 19.49 12.16 17.10 28.72 21.71 14.34 14.73 19.55 18.96 21.80 18.86 4.72 25.79 21.41 19.49 17.38 18.07 23.72 25.20 20.32 22.18 17.10 21.07 3.15 41.48 45.09 28.38 28.42 35.86 17.27 38.50 43.89 36.88 36.11 35.19 8.44 16.35 18.52 26.34 24.83 26.06 23.50 19.50 19.60 23.15 23.57 22.14 3.42 54.79 51.05 46.56 44.93 44.64 36.72 43.62 44.63 45.28 44.25 45.65 4.74 55.61 53.51 46.55 50.99 48.11 33.35 39.28 46.01 34.29 44.23 45.19 7.58 used liver mg microsomes for isolation (g) per g liver 1.92 11.38 1.76 12.89 1.45 12.43 1.19 11.87 1.49 9.77 1.48 10.75 1.42 12.50 1.6 10.29 1.52 15.52 1.31 15.04 12.24 1.89 2.52 7.74 1.04 11.69 1.8 9.50 2.23 12.88 1.48 14.67 1.05 13.66 1.16 12.70 1.67 11.71 1.34 14.15 1.79 12.18 12.09 2.12 1.76 14.66 1.98 10.81 1.97 9.90 1.5 11.59 1.39 13.00 1.78 13.32 1.67 15.09 1.35 15.05 2.18 10.17 1.14 15.00 12.86 2.10 3.1 13.38 3.33 13.54 3.33 8.52 3.53 8.05 3.89 9.22 1.72 10.04 3.21 11.99 3.43 12.80 3.64 10.13 3.19 11.32 10.90 2.00 1.71 9.56 1.51 12.27 2.32 11.35 1.47 16.89 2.14 12.18 2.17 10.83 1.32 14.77 1.65 11.88 1.62 14.29 1.86 12.67 12.67 2.13 3.49 15.70 4.36 11.71 4.1 11.36 4.17 10.78 4.13 10.81 3.13 11.73 2.78 15.69 3.57 12.50 4.17 10.86 3.85 11.49 12.26 1.88 3.85 14.44 3.86 13.86 3.27 14.24 3.92 13.01 3.7 13.00 3.48 9.58 3.27 12.01 3.36 13.69 2.92 11.74 3.48 12.71 12.83 1.45 AVERAGE SD group 1-7 12.26 1.97 Page A137 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Cholesterol-7--hydroxylase activity measurements in microsomes used per assay 420 g protein 20 min incubation internal standard 20 OH: 0.5 g per determination Mouse 1 13 21 31 45 51 61 2 14 22 32 46 52 62 3 17 23 33 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 7.489 7.764 8.344 7.459 7.921 8.048 7.859 8.072 8.614 8.284 8.279 7.708 7.956 8.246 8.569 8.034 8.103 8.551 8.177 7.462 8.615 8.176 7.997 8.678 7.153 8.014 7.760 7.119 8.211 8.341 7.684 7.654 8.062 8.155 8.529 8.412 8.674 8.281 9.215 7.851 8.260 8.138 9.023 9.148 8.783 7.856 7.946 7.525 7.842 7.885 8.094 7.994 7.866 7.808 area 7aOH 1.417 1.459 0.055 2.470 2.542 0.046 2.287 2.372 0.036 0.709 0.725 0.012 2.363 2.406 0.054 0.580 0.602 0.017 0.525 0.452 n.d. 2.048 2.039 0.036 0.912 0.988 0.018 1.032 1.302 0.026 0.389 0.369 n.d. 1.034 0.991 0.027 0.344 0.332 n.d. 0.414 0.408 n.d. 2.248 2.242 n.d. 2.554 2.519 0.055 2.020 2.034 0.031 0.878 0.863 n.d. g 7aOH formed 0.095 0.094 0.003 0.166 0.160 0.003 0.146 0.147 0.002 0.043 0.044 0.001 0.149 0.146 0.003 0.036 0.037 0.001 0.032 0.030 0.000 0.125 0.127 0.002 0.064 0.062 0.001 0.072 0.079 0.002 0.025 0.024 0.000 0.063 0.058 0.002 0.020 0.020 0.000 0.026 0.025 0.000 0.125 0.123 0.000 0.163 0.159 0.004 0.129 0.129 0.002 0.055 0.055 0.000 7 alpha hydroxylase activity (nmol/h/mg protein) 1.681 2.959 2.662 0.785 2.661 0.658 0.576 2.296 1.137 1.373 0.456 1.093 0.368 0.472 2.282 2.898 2.346 1.014 Page A138 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Mouse 41 53 63 4 11 24 34 42 54 64 6 12 25 35 43 55 65 5 15 26 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 8.173 7.392 7.547 7.601 8.694 8.113 8.815 8.171 8.490 8.332 8.091 8.957 8.472 7.915 8.828 8.352 8.133 8.018 7.148 8.432 6.831 8.519 8.106 8.352 8.875 7.996 8.907 8.561 9.027 9.222 8.466 8.414 8.950 7.978 8.371 9.549 8.359 7.939 7.754 7.600 8.216 7.859 8.187 7.860 7.683 9.259 8.480 8.305 7.459 8.254 8.066 8.428 8.346 7.805 8.127 8.333 8.348 7.974 7.537 8.761 area 7aOH 3.471 2.911 0.024 0.537 0.542 n.d. 0.556 0.583 n.d. 2.127 2.288 n.d. 1.862 1.780 0.029 1.753 1.984 n.d. 0.368 0.396 n.d. 2.052 1.990 n.d. 0.496 0.455 n.d. 0.678 0.653 n.d. 1.697 1.674 0.029 6.870 6.821 0.075 1.183 1.091 0.011 0.546 0.559 n.d. 2.021 1.895 0.020 0.619 0.595 0.024 1.066 1.116 n.d. 2.606 2.667 0.010 3.369 3.371 n.d. 2.884 2.681 0.043 g 7aOH formed 0.212 0.197 0.002 0.035 0.031 0.000 0.032 0.036 0.000 0.128 0.141 0.000 0.110 0.112 0.002 0.105 0.122 0.000 0.026 0.023 0.000 0.120 0.123 0.000 0.028 0.028 0.000 0.040 0.036 0.000 0.100 0.099 0.002 0.431 0.407 0.004 0.071 0.069 0.001 0.036 0.034 0.000 0.123 0.121 0.001 0.033 0.035 0.000 0.071 0.068 0.000 0.155 0.160 0.001 0.207 0.202 0.000 0.181 0.178 0.002 7 alpha hydroxylase activity (nmol/h/mg protein) 3.751 0.614 0.621 2.485 2.023 2.096 0.455 2.246 0.521 0.700 1.815 7.668 1.275 0.646 2.229 0.633 1.284 2.892 3.783 3.268 Page A139 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Mouse 36 44 56 66 7 16 27 37 47 57 67 8 18 28 38 48 58 68 9 19 29 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 7.543 7.412 8.080 8.523 8.450 7.586 8.189 9.455 7.871 9.543 8.293 8.583 6.707 8.792 8.695 7.664 8.129 9.510 9.979 9.559 9.372 8.482 9.335 7.948 8.375 8.607 9.266 7.959 10.692 8.987 8.115 9.262 8.761 4.538 4.178 4.434 3.873 4.184 4.643 8.343 9.026 9.171 3.989 3.669 4.843 4.001 4.331 5.012 3.910 4.155 5.027 4.100 4.005 4.765 4.192 4.391 4.288 4.084 4.075 4.527 4.149 3.944 4.428 area 7aOH 4.629 4.514 0.158 4.801 4.542 0.494 0.921 0.949 0.014 1.148 0.974 0.026 3.465 4.504 0.064 6.853 7.362 n.d. 2.836 2.618 n.d. 0.848 0.932 n.d. 4.765 4.853 0.083 0.895 1.034 0.006 0.890 0.924 0.028 2.079 2.064 0.049 1.823 1.740 0.032 4.317 4.463 0.031 0.463 0.409 n.d. 1.789 1.905 0.014 0.450 0.453 n.d. 0.821 0.866 0.037 2.296 2.352 0.023 2.755 2.820 n.d. 1.052 1.025 0.036 g 7aOH formed 0.307 0.305 0.010 0.282 0.269 0.033 0.056 0.050 0.001 0.060 0.059 0.000 0.258 0.256 0.004 0.447 0.453 0.000 0.142 0.137 0.000 0.050 0.050 0.000 0.284 0.282 0.004 0.056 0.048 0.000 0.055 0.050 0.000 0.229 0.247 0.006 0.235 0.208 0.003 0.259 0.247 0.002 0.058 0.056 0.000 0.224 0.220 0.001 0.058 0.055 0.000 0.100 0.108 0.004 0.274 0.268 0.003 0.337 0.346 0.000 0.127 0.130 0.004 7 alpha hydroxylase activity (nmol/h/mg protein) 5.466 4.482 0.967 1.098 4.684 8.312 2.577 0.923 5.149 0.960 0.967 4.295 4.031 4.642 1.051 4.071 1.035 1.852 4.954 6.311 2.296 Page A140 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Mouse 39 49 59 69 10 20 40 60 30 50 70 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 4.145 4.132 3.371 4.812 3.305 4.276 3.994 3.969 4.551 3.328 3.792 10.237 9.887 11.010 8.101 9.894 12.152 11.089 10.748 10.280 11.854 9.866 10.231 11.422 10.852 10.771 11.044 10.087 10.560 10.687 10.035 10.174 10.971 n.d.: not detectable area 7aOH 0.417 0.483 n.d. 1.356 0.979 n.d. 0.219 0.210 n.d. 0.710 0.735 n.d. 3.986 4.342 0.026 3.083 3.063 0.073 0.839 0.883 n.d. 0.951 0.893 0.040 2.471 2.346 n.d. 4.003 3.663 n.d. 1.536 1.606 n.d. g 7aOH formed 0.050 0.058 0.000 0.141 0.148 0.000 0.027 0.026 0.000 0.107 0.097 0.000 0.202 0.197 0.002 0.156 0.126 0.003 0.039 0.043 0.000 0.048 0.044 0.002 0.114 0.109 0.000 0.198 0.173 0.000 0.077 0.079 0.000 7 alpha hydroxylase activity (nmol/h/mg protein) 1.004 2.669 0.498 1.880 3.654 2.542 0.757 0.816 2.057 3.435 1.436 Page A141 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Group 1 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 7 AVERAGE SD mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 liver weight (g) 2.19 2.00 1.66 1.41 1.68 1.70 1.61 1.77 1.68 1.49 2.77 1.17 1.88 2.53 1.68 1.23 1.35 1.84 1.53 1.98 2.00 2.23 2.27 1.68 1.56 2.06 1.85 1.51 2.42 2.14 3.46 3.68 3.69 3.93 4.23 1.94 3.62 3.90 4.03 3.39 1.90 1.71 2.48 1.69 2.25 2.46 1.47 1.88 1.82 2.14 3.90 4.87 4.10 4.17 4.13 3.48 3.13 4.11 4.74 4.33 4.29 4.33 3.88 4.28 4.15 3.85 3.91 3.83 3.33 3.92 average mg microsomes/g liver 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 cholesterol 7 alpha hydroxylase activity (nmol/h/mg protein) per liver (nmol/h) 1.681 45.14 2.296 56.32 2.282 46.47 2.485 42.97 2.892 59.59 1.815 37.83 4.684 92.49 4.295 93.24 4.954 102.07 3.654 66.76 3.104 64.29 1.206 23.55 2.023 68.74 7.668 110.02 2.959 68.22 1.137 35.27 3.783 77.94 8.312 125.39 2.898 47.98 4.031 90.96 6.311 118.43 2.542 61.74 4.166 80.47 2.442 30.16 2.662 65.31 1.373 37.55 2.346 65.30 2.096 43.19 1.275 24.40 3.268 82.56 2.577 58.48 4.642 85.97 2.296 68.15 2.057 54.00 2.459 58.49 0.965 19.37 0.785 33.32 0.456 20.60 1.014 45.89 0.455 21.91 0.646 33.51 5.466 130.06 0.923 40.97 1.051 50.26 1.004 49.65 0.757 31.48 1.256 45.77 1.495 31.40 3.751 87.40 2.246 47.11 2.229 67.81 4.482 92.91 2.661 73.43 1.093 32.96 5.149 92.83 4.071 93.86 2.669 59.59 3.435 90.15 3.179 73.80 1.221 21.60 0.658 31.48 0.368 22.00 0.614 30.88 0.521 26.64 0.633 32.05 0.967 41.25 0.960 36.84 1.035 52.17 0.498 28.93 0.816 43.33 0.707 34.56 0.226 8.94 0.576 30.32 0.472 25.05 0.621 29.54 0.700 36.74 1.284 65.37 1.098 51.84 0.967 46.38 1.852 86.98 1.880 76.80 1.436 69.03 1.089 51.81 0.516 21.79 Page A142 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 HMG CoA reductase activity measurements internal standard mix 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 mouse# 1 13 21 31 45 51 61 2 14 22 32 46 52 3 17 23 33 53 4 11 24 34 42 54 64 6 12 25 35 blanc blanc 3H dpm (internal standard) 14802 15138 14416 16500 16182 18200 16197 16086 15302 17401 19233 18151 18408 16484 14705 17314 no sample 19044 17259 17962 16815 18678 17627 16776 18636 13567 12055 12101 13870 12322 13846 15599 13354 14885 17744 17935 15926 15147 17598 18276 17701 13443 16084 13092 12500 12547 12475 15313 15836 11436 13617 15941 12877 13025 17786 15984 16769 15482 14676 12984 14C dpm 222 207 199 294 310 278 437 321 187 224 296 360 192 169 132 159 172 124 137 205 265 140 158 226 146 118 92 143 138 116 144 197 241 231 266 190 172 221 316 187 156 260 209 127 116 114 145 119 86 104 110 196 254 405 279 378 214 162 118 14C dpm -blanc 82 67 59 154 170 138 297 181 47 84 156 220 52 29 -8 19 14C mevalonate formed (dpm) 169 135 125 284 320 231 558 342 93 147 247 369 86 54 -17 33 HMG CoA activity pmol/min/mg protein 5.111 4.083 3.776 8.611 9.692 6.995 16.917 10.381 2.834 4.454 7.483 11.182 2.606 1.623 -0.502 1.012 32 51 -16 -28 -3 -5 65 118 125 204 0 0 18 33 86 140 6 13 -22 -56 -48 -121 3 7 -2 -5 -24 -53 4 8 57 130 101 207 91 156 126 214 50 96 32 64 81 140 176 293 47 81 16 36 120 227 69 160 -13 -32 -24 -58 -26 -63 5 10 -21 -40 -54 -144 -36 -80 -30 -57 56 132 114 266 265 453 139 247 238 403 74 136 1.550 -0.855 -0.154 3.566 6.174 0.000 0.990 4.257 0.408 -1.684 -3.659 0.200 -0.150 -1.599 0.237 3.938 6.260 4.731 6.481 2.896 1.949 4.246 8.885 2.450 1.098 6.883 4.862 -0.959 -1.765 -1.923 0.301 -1.223 -4.356 -2.439 -1.736 4.012 8.075 13.746 7.482 12.211 4.112 mean 4.60 6.19 8.34 13.65 3.64 9.33 2.11 0.26 1.55 0.00 4.87 0.49 2.33 0.00 0.02 0.00 5.10 5.61 2.42 6.57 1.77 5.87 0.00 0.00 0.00 0.00 6.04 10.61 8.16 Input internal standard (dpm): average mix 1 30176 29765 31362 30434 mix 2 28810 28307 28031 28383 14C dpm HMG CoA mix: average 58465 57242 55833 57180 11436 per 5 l mix per l mix HMG CoA mix: Specific activity 14C HMGCoA: incubation time: liver microsomes: 2.6 4398.46 30.0 0.250 mmol/l HMG CoA dpm/nmol min mg Page A143 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 mouse# 43 55 65 5 15 26 36 44 66 7 16 27 37 blanc blanc 3H dpm (internal standard) 16023 14382 15354 16171 15593 15352 13631 12996 13907 11471 10075 12964 13289 16215 17106 14863 14921 14914 14706 15107 13433 14340 14736 15759 16357 15923 14228 11628 Input internal standard (dpm): average 27932 27694 28968 28198 14C dpm 252 192 261 211 451 136 140 139 98 109 97 93 139 173 402 338 155 168 195 321 191 287 142 154 204 179 194 144 14C dpm -blanc 83 23 92 42 282 -33 -29 -30 -71 -60 -72 -76 -30 4 233 169 -14 -1 26 152 22 118 -27 -15 35 10 14C mevalonate formed (dpm) 146 45 169 73 510 -61 -60 -65 -144 -147 -202 -165 -64 7 384 321 -26 -2 50 284 46 232 -52 -27 60 18 HMG CoA act. pmol/min.mg eiwit 4.237 1.308 4.901 2.124 14.792 -1.758 -1.740 -1.888 -4.176 -4.278 -5.845 -4.795 -1.846 0.202 11.141 9.300 -0.767 -0.055 1.446 8.230 1.340 6.731 -1.499 -0.779 1.750 0.514 mean 2.77 3.51 6.52 0.00 0.00 0.00 0.00 10.22 0.00 4.84 0.00 0.00 1.13 14C HMG CoA mix: average 58407 55714 65147 59756 11951.2 per 5 l mix per l mix HMG CoA mix: Specific activity 14C HMGCoA: incubation time: liver microsomes: 2.6 mmol/l HMG CoA 4596.62 dpm/nmol 30.0 min 0.250 mg mouse# 67 8 9 19 29 39 49 59 50 70 blanc blanc 3H dpm (internal standard) 16112 16092 14295 17696 17286 16982 14651 19083 16755 16504 17225 17750 18651 15076 15831 16644 16807 14453 16679 15823 16121 16049 Input internal standard (dpm): average 27956 27723 28126 27935 14C dpm 284 180 129 164 379 317 145 113 142 115 247 218 209 152 159 154 195 321 191 287 95 72 14C dpm -blanc 201 97 46 81 296 234 62 30 59 32 164 135 126 69 76 71 112 238 108 204 14C mevalonate formed (dpm) 348 168 89 127 478 384 117 43 98 53 265 212 188 127 133 118 185 459 180 359 HMG CoA act. pmol/min.mg eiwit 9.646 4.648 2.467 3.526 13.251 10.658 3.254 1.198 2.706 1.479 7.358 5.874 5.216 3.522 3.697 3.283 5.142 12.738 4.996 9.969 mean 7.15 3.00 11.95 2.23 2.09 6.62 4.37 3.49 8.94 7.48 14C HMG CoA mix: average 63731 60065 63603 62466 12493.27 per 5 l mix per l mix HMG CoA mix: Specific activity 14C HMGCoA: incubation time: liver microsomes: 2.6 mmol/l HMG CoA 4805.1 dpm/nmol 30.0 min 0.250 mg Page A144 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 mouse# 18 28 38 48 58 68 69 10 20 40 30 62 41 63 56 47 blanc 3H dpm (internal standard) 18621 18163 16885 17274 15495 18485 15821 17041 19175 17310 16607 14530 16586 14581 15189 15445 16021 16747 15999 14205 20405 19844 16228 14978 15343 13316 15166 16339 17431 14614 15456 15823 16766 Input internal standard (dpm): average 32959 31221 31297 31826 14C dpm 513 361 265 260 682 691 341 312 412 318 241 348 352 360 275 241 224 235 404 355 591 403 328 218 233 317 214 302 407 514 650 287 164 14C dpm -blanc 349 197 101 96 518 527 177 148 248 154 77 184 188 196 111 77 60 71 240 191 427 239 164 54 69 153 50 138 243 350 486 123 14C mevalonate formed (dpm) 596 345 190 177 1064 907 356 276 412 283 148 403 361 428 233 159 119 135 477 428 666 383 322 115 143 366 105 269 444 762 1001 247 HMG CoA act. pmol/min.mg eiwit 17.513 10.135 5.589 5.193 31.238 26.640 10.454 8.115 12.085 8.313 4.333 11.833 10.592 12.561 6.829 4.658 3.499 3.962 14.017 12.564 19.554 11.254 9.443 3.369 4.202 10.736 3.081 7.892 13.026 22.379 29.382 7.264 mean 13.82 5.39 28.94 9.28 10.20 8.08 11.58 5.74 3.73 13.29 15.40 6.41 7.47 5.49 17.70 18.32 14C HMG CoA mix: average 61119 60406 55583 59036 11807.2 per 5 l mix per l mix HMG CoA mix: Specific activity 14C HMGCoA: incubation time: liver microsomes: 2.6 mmol/l HMG CoA 4541.23 dpm/nmol 30.0 min 0.250 mg Page A145 of A172 Appendix to TNO-report of 3M-02 Final version Group mouse# 1 1 1 2 1 3 1 4 1 5 1 6 1 7 1 8 1 9 1 10 AVERAGE SD 2 11 2 12 2 13 2 14 2 15 2 16 2 17 2 18 2 19 2 20 AVERAGE SD 3 21 3 22 3 23 3 24 3 25 3 26 3 27 3 28 3 29 3 30 AVERAGE SD 4 31 4 32 4 33 4 34 4 35 4 36 4 37 4 38 4 39 4 40 AVERAGE SD 5 41 5 42 5 43 5 44 5 45 5 46 5 47 5 48 5 49 5 50 AVERAGE SD 6 51 6 52 6 53 6 54 6 55 6 56 6 57 6 58 6 59 6 60 AVERAGE SD 7 61 7 62 7 63 7 64 7 65 7 66 7 67 7 68 7 69 7 70 AVERAGE SD liver weight (g) 2.19 2.00 1.66 1.41 1.68 1.70 1.61 1.77 1.68 1.49 average mg microsomes/g liver 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 12.26 2.77 12.26 1.17 12.26 1.88 12.26 2.53 12.26 1.68 12.26 1.23 12.26 1.35 12.26 1.84 12.26 1.53 12.26 1.98 12.26 2.00 12.26 2.23 12.26 2.27 12.26 1.68 12.26 1.56 12.26 2.06 12.26 1.85 12.26 1.51 12.26 2.42 12.26 2.14 12.26 3.46 12.26 3.68 12.26 3.69 12.26 3.93 12.26 4.23 12.26 1.94 12.26 3.62 12.26 3.90 12.26 4.03 12.26 3.39 12.26 1.90 12.26 1.71 12.26 2.48 12.26 1.69 12.26 2.25 12.26 2.46 12.26 1.47 12.26 1.88 12.26 1.82 12.26 2.14 12.26 3.90 12.26 4.87 12.26 4.10 12.26 4.17 12.26 4.13 12.26 3.48 12.26 3.13 12.26 4.11 12.26 4.74 12.26 4.33 12.26 4.29 12.26 4.33 12.26 3.88 12.26 4.28 12.26 4.15 12.26 3.85 12.26 3.91 12.26 3.83 12.26 3.33 12.26 3.92 12.26 HMG CoA reductase activity (pmol/min/mg protein) per liver (pmol/min) 4.60 123.43 0.26 6.26 0.00 0.00 2.42 41.88 0.00 0.00 0.00 0.00 4.84 95.49 3.00 65.03 11.95 246.23 5.74 104.92 3.28 68.32 3.77 78.41 6.57 222.96 6.04 86.69 6.19 142.75 1.55 48.08 0.00 0.00 0.00 0.00 0.02 0.41 13.82 311.85 2.23 41.76 3.73 90.56 4.02 94.51 4.33 104.24 8.34 204.59 0.00 0.00 0.00 0.00 1.77 36.54 10.61 203.00 0.00 0.00 0.00 0.00 5.39 99.81 2.09 62.10 15.40 404.15 4.36 101.02 5.44 133.13 13.65 578.98 4.87 219.73 5.10 230.67 5.87 282.96 8.16 423.27 0.00 0.00 1.13 50.24 28.94 1383.68 6.62 326.87 13.29 552.38 8.76 404.88 8.36 392.39 7.47 173.99 0.00 0.00 2.77 84.30 10.22 211.77 3.64 100.51 0.49 14.93 18.32 330.22 9.28 214.00 4.37 97.48 8.94 234.56 6.55 146.18 5.51 104.48 9.33 446.23 2.33 139.28 5.61 281.81 0.00 0.00 3.51 177.86 17.70 755.28 6.15 236.05 10.20 513.93 3.49 202.82 6.69 355.05 6.50 310.83 5.01 216.26 2.11 111.22 6.41 340.07 5.49 260.98 0.00 0.00 6.52 331.58 0.00 0.00 7.15 342.61 8.08 379.53 11.58 472.60 7.48 359.61 5.48 259.82 3.71 165.03 04-12-2006 Page A146 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 micromes mg/g liver 7-alpha act. 7-alpha act. per liver HMG Co A act. HMG Co A act. per liver micromes mg/g liver 7-alpha act. 7-alpha act. per liver HMG Co A act. HMG Co A act. per liver micromes mg/g liver 7-alpha act. 7-alpha act. per liver HMG Co A act. HMG Co A act. per liver micromes; mg per group 1 vs 2 nn p 10 10 0.971 10 10 0.353 10 10 0.190 10 10 0.631 10 10 0.739 group 2 vs 4 nn p 10 10 0.218 10 10 <0.001 10 10 0.009 10 10 0.190 10 10 0.015 group 3 vs 7 nn p 10 10 0.796 10 10 0.001 10 10 0.631 10 10 0.436 10 10 0.063 g liver, 7-alpha-hydroxylase activity and HMG CoA group 1 vs 3 group 1 vs 4 group 1 vs 5 nn p nn p nn p 10 10 0.529 10 10 0.143 10 10 0.739 10 10 0.393 10 10 0.002 10 10 0.912 10 10 0.684 10 10 0.035 10 10 0.353 10 10 1.000 10 10 0.029 10 10 0.165 10 10 1.000 10 10 0.007 10 10 0.105 group 2 vs 5 group 2 vs 6 group 2 vs 7 nn p nn p nn p 10 10 0.579 10 10 0.579 10 10 0.315 10 10 0.529 10 10 <0.001 10 10 <0.001 10 10 0.684 10 10 <0.001 10 10 0.043 10 10 0.247 10 10 0.247 10 10 0.315 10 10 0.247 10 10 0.015 10 10 0.035 group 4 vs 5 group 4 vs 6 group 4 vs 7 nn p nn p nn p 10 10 0.063 10 10 0.029 10 10 0.029 10 10 0.002 10 10 0.529 10 10 0.529 10 10 0.015 10 10 0.353 10 10 0.353 10 10 0.631 10 10 0.684 10 10 0.579 10 10 0.043 10 10 0.631 10 10 0.684 reductase activity, group 1 vs 6 nn p 10 10 0.971 10 10 <0.001 10 10 <0.001 10 10 0.105 10 10 0.003 group 3 vs 4 nn p 10 10 0.035 10 10 0.002 10 10 0.052 10 10 0.218 10 10 0.011 group 5 vs 6 nn p 10 10 0.481 10 10 <0.001 10 10 <0.001 10 10 0.912 10 10 0.052 p values Mann-Whitney U-test group 1 vs 7 group 2 vs 3 nn p nn p 10 10 0.315 10 10 0.436 10 10 <0.001 10 10 0.105 10 10 0.247 10 10 0.089 10 10 0.165 10 10 0.796 10 10 0.019 10 10 0.739 group 3 vs 5 group 3 vs 6 nn p nn p 10 10 0.912 10 10 0.738 10 10 0.190 10 10 <0.001 10 10 0.075 10 10 0.005 10 10 0.280 10 10 0.218 10 10 0.190 10 10 0.023 group 5 vs 7 group 6 vs 7 nn p nn p 10 10 0.971 10 10 0.143 10 10 <0.001 10 10 0.143 10 10 0.029 10 10 0.123 10 10 0.684 10 10 0.912 10 10 0.063 10 10 0.853 p<0.05 Page A147 of A172 Appendix to TNO-report of 3M-02 Final version H2. Cholesterol-27-hydroxylase activity in mitochondria Protein concentration isolated mitochondria Mitochondria mouse 1 13 21 31 45 51 61 2 14 22 32 46 52 62 3 17 23 33 41 53 63 4 11 24 34 42 54 64 6 12 25 35 43 55 65 5 15 26 36 44 56 66 7 16 27 37 47 57 67 8 18 28 38 48 58 68 9 19 29 39 49 59 69 10 20 40 60 30 50 70 measured prot conc (mg/ml) well 1 well 2 0.772 0.8 0.699 0.699 0.848 0.908 1.508 1.499 0.925 0.973 0.784 0.813 0.865 0.881 1.143 1.162 0.918 0.931 0.945 1.022 1.192 1.104 1.043 1.062 1.26 1.236 1.201 1.204 0.81 0.81 0.57 0.54 0.915 0.949 1.036 1.092 0.699 0.655 1.033 0.942 0.898 0.839 0.576 0.587 0.726 0.672 0.969 0.959 1.532 1.43 0.784 0.852 0.98 0.997 1.201 1.28 0.699 0.658 0.383 0.359 0.931 0.861 1.37 1.235 0.842 0.845 0.945 0.976 0.949 0.901 0.649 0.637 0.648 0.635 0.775 0.799 0.678 0.705 0.757 0.757 0.84 0.843 1.031 0.998 0.754 0.714 0.467 0.443 0.876 0.886 1.171 1.1 0.554 0.554 0.847 0.809 0.578 0.596 0.616 0.596 0.697 0.7 0.728 0.745 1.808 1.903 1.086 0.85 1.261 1.147 0.734 0.766 0.616 0.611 0.691 0.659 0.847 0.847 1.255 1.251 0.76 0.751 0.85 0.796 0.751 0.728 0.837 0.821 0.84 0.809 1.344 1.34 0.769 0.827 0.642 0.598 0.987 0.965 1.039 0.926 dilution 50 50 50 50 50 100 100 50 50 50 100 50 100 100 50 50 50 100 50 100 100 50 50 50 100 50 100 100 50 50 50 100 50 100 100 50 50 50 50 50 100 100 50 50 50 100 50 100 100 50 50 50 100 50 100 100 50 50 50 100 50 100 100 50 50 100 100 50 50 100 Protein conc (mg/ml) 39.30 34.95 43.90 75.18 47.45 79.85 87.30 57.63 46.23 49.18 114.80 52.63 124.80 120.25 40.50 27.75 46.60 106.40 33.85 98.75 86.85 29.08 34.95 48.20 148.10 40.90 98.85 124.05 33.93 18.55 44.80 130.25 42.18 96.05 92.50 32.15 32.08 39.35 34.58 37.85 84.15 101.45 36.70 22.75 44.05 113.55 27.70 82.80 58.70 30.30 34.93 36.83 185.55 48.40 120.40 75.00 30.68 33.75 42.35 125.30 37.78 82.30 73.95 41.45 41.23 134.20 79.80 31.00 48.80 98.25 04-12-2006 Page A148 of A172 Appendix to TNO-report of 3M-02 Final version Amount isolated mitochondria group# 1 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 7 AVERAGE SD mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 Protein conc mitochondria (mg/ml) 39.30 57.63 40.50 29.08 32.15 33.93 36.70 30.30 30.68 41.45 estimated l microsomes 600 600 600 600 600 600 600 600 600 600 34.95 600 18.55 600 34.95 600 46.23 600 32.08 600 22.75 600 27.75 600 34.93 600 33.75 600 41.23 600 43.90 600 49.18 600 46.60 600 48.20 600 44.80 600 39.35 600 44.05 600 36.83 600 42.35 600 31.00 600 75.18 700 114.80 700 106.40 700 148.10 700 130.25 700 34.58 600 113.55 700 185.55 700 125.30 700 134.20 700 33.85 600 40.90 600 42.18 600 37.85 600 47.45 600 52.63 600 27.70 600 48.40 600 37.78 600 48.80 600 79.85 700 124.80 700 98.75 700 98.85 700 96.05 700 84.15 700 82.80 700 120.40 700 82.30 700 79.80 700 87.30 700 120.25 700 86.85 700 124.05 700 92.50 700 101.45 700 58.70 700 75.00 700 73.95 700 98.25 700 isolated mitochondria (mg) 23.58 34.58 24.30 17.45 19.29 20.36 22.02 18.18 18.41 24.87 20.97 11.13 20.97 27.74 19.25 13.65 16.65 20.96 20.25 24.74 26.34 29.51 27.96 28.92 26.88 23.61 26.43 22.10 25.41 18.60 52.62 80.36 74.48 103.67 91.18 20.75 79.49 129.89 87.71 93.94 20.31 24.54 25.31 22.71 28.47 31.58 16.62 29.04 22.67 29.28 55.90 87.36 69.13 69.20 67.24 58.91 57.96 84.28 57.61 55.86 61.11 84.18 60.80 86.84 64.75 71.02 41.09 52.50 51.77 68.78 64.28 14.24 used liver for isolation (g) 1.92 1.76 1.45 1.19 1.49 1.48 1.42 1.6 1.52 1.31 2.52 1.04 1.8 2.23 1.48 1.05 1.16 1.67 1.34 1.79 1.76 1.98 1.97 1.5 1.39 1.78 1.67 1.35 2.18 1.14 3.1 3.33 3.33 3.53 3.89 1.72 3.21 3.43 3.64 3.19 1.71 1.51 2.32 1.47 2.14 2.17 1.32 1.65 1.62 1.86 3.49 4.36 4.1 4.17 4.13 3.13 2.78 3.57 4.17 3.85 3.85 3.86 3.27 3.92 3.7 3.48 3.27 3.36 2.92 3.48 mg mitochondria per g liver 12.28 19.64 16.76 14.66 12.95 13.75 15.51 11.36 12.11 18.98 14.80 2.89 8.32 10.70 11.65 12.44 13.00 13.00 14.35 12.55 15.11 13.82 12.49 1.94 14.97 14.90 14.19 19.28 19.34 13.26 15.83 16.37 11.66 16.32 15.61 2.42 16.98 24.13 22.37 29.37 23.44 12.06 24.76 37.87 24.10 29.45 24.45 7.03 11.88 16.25 10.91 15.45 13.30 14.55 12.59 17.60 13.99 15.74 14.23 2.09 16.02 20.04 16.86 16.59 16.28 18.82 20.85 23.61 13.82 14.51 17.74 3.05 15.87 21.81 18.59 22.15 17.50 20.41 12.57 15.63 17.73 19.76 18.20 2.99 AVERAGE SD group 1-3,5 14.28 2.55 AVERAGE SD group 4 24.45 7.03 AVERAGE SD group 6,7 17.97 2.95 04-12-2006 Page A149 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Cholesterol-27-hydroxylase activity measurements used per assay 420 g protein 20 min incubation internal standard 20 OH: 0.5 g per determination Mouse 1 13 21 31 45 51 61 2 14 22 32 46 52 62 3 17 23 33 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 10.798 10.39 7.98 11.806 12.439 8.742 11.5 11.782 8.867 9.271 10.042 8.477 12.431 12.184 8.816 10.388 10.595 8.259 9.893 10.305 8.75 9.624 10.36 8.95 11.022 11.6 9.308 10.919 10.512 9.52 8.987 9.298 8.538 10.614 10.223 9.715 9.353 10.539 8.851 10.024 9.352 8.92 12.459 11.384 9.006 10.383 14.276 9.026 9.646 10.293 7.233 8.216 8.156 7.185 area 27OH 13.936 13.174 0.136 20.962 19.566 0.125 19.885 16.14 0.126 8.456 9.881 0.038 18.514 17.151 0.07 10.217 11.997 0.113 11.615 11.839 0.068 11.398 13.32 n.a. 14.253 14.61 0.023 15.555 16.346 0.074 8.043 8.372 0.023 15.727 15.178 0.048 9.827 9.649 0.071 6.711 6.677 n.a. 19.765 15.393 n.a. 12.005 18.144 n.a. 14.544 14.524 n.a. 7.206 6.744 0.249 g 27 OH formed 0.645 0.634 0.009 0.888 0.786 0.007 0.865 0.685 0.007 0.456 0.492 0.002 0.745 0.704 0.004 0.492 0.566 0.007 0.587 0.574 0.004 0.592 0.643 0.000 0.647 0.630 0.001 0.712 0.777 0.004 0.447 0.450 0.001 0.741 0.742 0.002 0.525 0.458 0.004 0.335 0.357 0.000 0.793 0.676 0.000 0.578 0.635 0.000 0.754 0.706 0.000 0.439 0.413 0.017 27- hydroxylase activity (nmol/h/mg protein) 11.659 15.332 14.181 8.715 13.306 9.645 10.656 11.407 11.766 13.689 8.267 13.654 9.007 6.389 13.571 11.209 13.480 7.549 Page A150 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Mouse 41 53 63 4 11 24 34 42 54 64 6 12 25 35 43 55 65 5 15 26 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 10.176 9.541 7.88 7.856 7.979 7.279 10.998 8.797 7.705 9.576 10.192 7.719 11.137 9.797 8.556 10.407 9.642 6.778 6.577 9.528 7.252 9.538 9.074 7.471 8.887 8.267 7.436 8.585 8.945 6.454 11.823 8.846 8.775 9.224 9.098 9.234 10.72 9.542 8.864 9.856 10.84 8.053 10.178 10.114 7.765 8.696 8.713 8.251 9.579 9.473 8.811 9.569 9.56 8.442 10.061 10.559 9.105 9.863 9.469 8.926 area 27OH 4.943 4.889 n.a. 4.981 4.873 n.a. 7.897 6.181 0.086 11.39 12.068 n.a. 8.145 7.601 0.038 11.002 11.512 n.a. 3.674 5.095 n.a. 11.731 10.387 0.027 6.229 5.477 n.a. 6.457 6.783 n.a. 13.325 9.816 0.057 1.784 1.721 n.a. 7.132 7.327 0.031 5.695 4.8 n.a. 12.222 10.741 n.a. 5.552 5.349 0.032 6.842 7.217 0.056 7.618 7.922 0.004 8.488 7.934 n.a. 7.929 8.269 n.a. g 27 OH formed 0.243 0.256 0.000 0.317 0.305 0.000 0.359 0.351 0.006 0.595 0.592 0.000 0.366 0.388 0.002 0.529 0.597 0.000 0.279 0.267 0.000 0.615 0.572 0.002 0.350 0.331 0.000 0.376 0.379 0.000 0.564 0.555 0.003 0.097 0.095 0.000 0.333 0.384 0.000 0.289 0.221 0.000 0.600 0.531 0.000 0.319 0.307 0.002 0.357 0.381 0.003 0.398 0.414 0.000 0.422 0.376 0.000 0.402 0.437 0.000 27- hydroxylase activity (nmol/h/mg protein) 4.610 5.749 6.458 10.962 6.920 10.396 5.049 10.933 6.297 6.976 10.270 1.767 6.619 4.714 10.450 5.748 6.758 7.499 7.366 7.746 Page A151 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Mouse 36 44 56 66 7 16 27 37 47 57 67 8 18 28 38 48 58 68 9 19 29 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 9.147 9.469 7.56 9.972 11.897 8.394 9.607 9.685 8.852 9.829 10.203 9.701 10.39 9.963 9.004 9.887 9.808 8.749 10.948 11.307 9.47 10.004 9.859 8.538 9.054 10.166 9.566 10.482 9.237 9.439 9.185 9.321 8.555 10.123 10.395 8.227 9.534 9.641 7.802 10.422 10.894 7.384 9.189 8.998 8.56 10.497 10.009 8.178 8.418 9.913 8.293 9.401 10.176 8.704 8.909 9.904 7.036 6.336 8.809 6.461 6.725 9.786 4.961 area 27OH 4.853 4.856 0.022 8.19 8.239 n.a. 5.319 4.876 0.035 7.898 7.893 0.042 6.813 7.322 n.a. 6.158 5.884 n.a. 8.509 8.456 0.018 6.283 6.029 n.a. 6.579 8.773 n.a. 5.812 5.663 n.a. 8.637 10.313 0.041 14.048 11.26 n.a. 9.63 8.353 n.a. 10.021 12.138 n.a. 3.667 3.925 n.a. 11.218 9.378 0.01 4.05 4.887 n.a. 6.674 6.558 n.a. 7.488 10.247 n.a. 5.573 7.57 n.a. 5.613 7.53 0.002 g 27 OH formed 0.265 0.256 0.001 0.411 0.346 0.000 0.277 0.252 0.002 0.402 0.387 0.002 0.328 0.367 0.000 0.311 0.300 0.000 0.389 0.374 0.001 0.314 0.306 0.000 0.363 0.431 0.000 0.277 0.307 0.000 0.470 0.553 0.002 0.694 0.542 0.000 0.505 0.433 0.000 0.481 0.557 0.000 0.200 0.218 0.000 0.534 0.468 0.001 0.241 0.246 0.000 0.355 0.322 0.000 0.420 0.517 0.000 0.440 0.430 0.000 0.417 0.385 0.000 27- hydroxylase activity (nmol/h/mg protein) 4.792 6.991 4.846 7.244 6.422 5.647 7.026 5.725 7.341 5.392 9.408 11.412 8.666 9.586 3.858 9.251 4.499 6.255 8.660 8.031 7.405 Page A152 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Mouse 39 49 59 69 10 20 40 60 30 50 70 energy generating mix added + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix + NADPH mix + NADPH mix - NADPH mix area 20 OH 8.832 9.279 8.084 9.633 11.234 7.927 9.799 9.597 9.087 9.836 9.96 1.235 9.549 10.004 7.526 9.543 9.019 8.008 7.696 8.096 7.695 8.639 8.551 8.104 8.271 8.406 7.768 9.353 9.445 7.913 9.568 8.325 7.941 area 27OH 4.203 4.385 n.a. 6.413 7.141 n.a. 4.515 4.761 n.a. 6.674 6.877 n.a. 8.229 9.15 n.a. 7.346 7.444 0.031 2.057 2.046 0.01 3.465 3.757 0.047 5.462 5.704 n.a. 8.537 7.619 n.a. 4.626 4.716 n.a. g 27 OH formed 0.238 0.236 0.000 0.333 0.318 0.000 0.230 0.248 0.000 0.339 0.345 0.000 0.431 0.457 0.000 0.385 0.413 0.002 0.134 0.126 0.001 0.201 0.220 0.003 0.330 0.339 0.000 0.456 0.403 0.000 0.242 0.283 0.000 27- hydroxylase activity (nmol/h/mg protein) 4.380 6.010 4.419 6.322 8.204 7.331 2.390 3.828 6.184 7.941 4.849 Page A153 of A172 Appendix to TNO-report of 3M-02 Final version Group 1 1 1 1 1 1 1 1 1 1 AVERAGE SD 2 2 2 2 2 2 2 2 2 2 AVERAGE SD 3 3 3 3 3 3 3 3 3 3 AVERAGE SD 4 4 4 4 4 4 4 4 4 4 AVERAGE SD 5 5 5 5 5 5 5 5 5 5 AVERAGE SD 6 6 6 6 6 6 6 6 6 6 AVERAGE SD 7 7 7 7 7 7 7 7 7 7 AVERAGE SD Mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 mouse# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 liver weight (g) 2.19 2.00 1.66 1.41 1.68 1.70 1.61 1.77 1.68 1.49 2.77 1.17 1.88 2.53 1.68 1.23 1.35 1.84 1.53 1.98 2.00 2.23 2.27 1.68 1.56 2.06 1.85 1.51 2.42 2.14 3.46 3.68 3.69 3.93 4.23 1.94 3.62 3.90 4.03 3.39 1.90 1.71 2.48 1.69 2.25 2.46 1.47 1.88 1.82 2.14 3.90 4.87 4.10 4.17 4.13 3.48 3.13 4.11 4.74 4.33 4.29 4.33 3.88 4.28 4.15 3.85 3.91 3.83 3.33 3.92 average mg mitochondria/g liver 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 24.45 24.45 24.45 24.45 24.45 24.45 24.45 24.45 24.45 24.45 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 14.28 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 17.97 cholesterol 27 hydroxylase activity (nmol/h/mg protein) per liver (nmol/h) 11.66 364.7 11.41 325.9 13.57 321.8 10.96 220.8 7.50 180.0 10.27 249.4 6.42 147.7 11.41 288.5 8.66 207.8 8.20 174.6 10.01 248.1 2.22 73.9 6.92 273.8 1.77 29.5 15.33 411.7 11.77 425.2 7.37 176.8 5.65 99.2 11.21 216.1 8.67 227.8 8.03 175.5 7.33 207.3 8.40 224.3 3.70 123.2 14.18 405.1 13.69 436.0 13.48 437.1 10.40 249.5 6.62 147.5 7.75 227.9 7.03 185.6 9.59 206.8 7.40 255.9 6.18 189.0 9.63 274.0 3.14 109.9 8.72 737.3 8.27 743.8 7.55 681.1 5.05 485.2 4.71 487.5 4.79 227.3 5.72 506.7 3.86 367.9 4.38 431.6 2.39 198.1 5.54 486.7 2.03 192.9 4.61 125.1 10.93 267.0 10.45 370.2 6.99 168.8 13.31 427.6 13.65 479.8 7.34 154.1 9.25 248.4 6.01 156.2 7.94 242.7 9.05 264.0 3.02 123.5 9.65 676.0 9.01 788.2 5.75 423.5 6.30 471.8 5.75 426.6 4.85 303.0 5.39 303.3 4.50 332.3 4.42 376.4 3.83 297.8 5.94 439.9 1.93 167.2 10.66 821.5 6.39 497.1 6.46 450.3 6.98 536.5 6.76 504.0 7.24 501.1 9.41 661.0 6.25 430.5 6.32 378.3 4.85 341.6 7.13 512.2 1.68 140.0 04-12-2006 Page A154 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 mitochondria mg/g liver 27 hydroxylase act. 27 hydroxylase act. per liver mitochondria mg/g liver 27 hydroxylase act. 27 hydroxylase act. per liver mitochondria mg/g liver 27 hydroxylase act. 27 hydroxylase act. per liver mitochondria; mg per g liver and 27-hydroxylase activity, p values Mann-Whitney U-test group 1 vs 2 group 1 vs 3 group 1 vs 4 group 1 vs 5 group 1 vs 6 group 1 vs 7 nn p nn p nn p nn p nn p nn p 10 10 0.165 10 10 0.436 10 10 0.002 10 10 0.796 10 10 0.043 10 10 0.019 10 10 0.247 10 10 0.631 10 10 0.003 10 10 0.353 10 10 0.001 10 10 0.004 10 10 0.579 10 10 0.684 10 10 0.001 10 10 0.971 10 10 0.002 10 10 <0.001 group 2 vs 4 group 2 vs 5 group 2 vs 6 group 2 vs 7 group 3 vs 4 group 3 vs 5 nn p nn p nn p nn p nn p nn p 10 10 <0.001 10 10 0.075 10 10 <0.001 10 10 <0.001 10 10 0.002 10 10 0.218 10 10 0.075 10 10 0.796 10 10 0.063 10 10 0.165 10 10 0.005 10 10 0.684 10 10 0.003 10 10 0.631 10 10 0.002 10 10 <0.001 10 10 0.015 10 10 0.739 group 3 vs 7 group 4 vs 5 group 4 vs 6 group 4 vs 7 group 5 vs 6 group 5 vs 7 nn p nn p nn p nn p nn p nn p 10 10 0.052 10 10 0.001 10 10 0.009 10 10 0.011 10 10 0.011 10 10 0.005 10 10 0.043 10 10 0.015 10 10 0.529 10 10 0.075 10 10 0.019 10 10 0.123 10 10 0.001 10 10 0.009 10 10 0.393 10 10 0.796 10 10 0.009 10 10 <0.001 group 2 vs 3 nn p 10 10 0.005 10 10 0.579 10 10 0.353 group 3 vs 6 nn p 10 10 0.123 10 10 0.003 10 10 0.019 group 6 vs 7 nn p 10 10 0.739 10 10 0.035 10 10 0.089 p<0.05 Page A155 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 I. Measurements in vitro experiment (rat hepatocytes) Experiment 1 Protein concentration determination sample 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 Measured protein conc (mg/ml) 97.5 90.6 119.5 125.5 172.9 140.5 155.8 180.3 167.7 177.1 131.6 121.2 139.7 114.5 257.5 214.7 176 175 155.8 141.6 155.8 122.1 167.7 154.8 129 127.2 136.1 141.5 170.8 177.1 172.9 185.7 168.7 155.6 177.1 165.7 151.9 150.9 188.9 183.5 190.1 229.9 168.7 162.7 167.7 157.7 229.9 218.4 150.9 146.2 127.2 133.4 172.9 151.9 170.8 185.7 170.8 178.1 163.7 170.8 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 average protein conc (mg/ml) 376.2 490.0 626.8 672.2 689.6 505.6 508.4 944.4 702.0 594.8 555.8 645.0 512.4 555.2 695.8 717.2 648.6 685.6 605.6 744.8 840.0 662.8 650.8 896.6 594.2 521.2 649.6 713.0 697.8 669.0 g protein per well 125.4 163.3 208.9 224.1 229.9 168.5 169.5 314.8 234.0 198.3 185.3 215.0 170.8 185.1 231.9 239.1 216.2 228.5 201.9 248.3 280.0 220.9 216.9 298.9 198.1 173.7 216.5 237.7 232.6 223.0 sample 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 Measured protein conc (mg/ml) 134.3 127.2 149.9 131.6 202.7 195.7 194.1 216.8 188.7 203.7 173.8 161.3 194.1 216.8 188.7 203.8 173.9 161.3 168.6 173.9 140.8 143.8 139.7 144.8 155.1 147.9 162.4 164.4 164.4 160.3 179.2 169.7 198.4 167.6 209.1 195.1 207 216.8 239.9 255.6 241.1 249.9 182.33 132.7 201.6 195.1 212.4 221.2 210.4 193 211.4 243.3 208.1 205.9 239.9 223.4 213.5 211.4 198.4 181.3 129.7 133.7 156.1 181.3 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 average protein conc (mg/ml) 523.0 563.0 796.8 821.8 784.8 670.2 821.8 785.0 670.4 685.0 569.2 569.0 606.0 653.6 649.4 697.8 732.0 808.4 847.6 991.0 982.0 630.1 793.4 867.2 806.8 909.4 828.0 926.6 849.8 759.4 526.8 674.8 g protein per well 174.3 187.7 265.6 273.9 261.6 223.4 273.9 261.7 223.5 228.3 189.7 189.7 202.0 217.9 216.5 232.6 244.0 269.5 282.5 330.3 327.3 210.0 264.5 289.1 268.9 303.1 276.0 308.9 283.3 253.1 175.6 224.9 Page A156 of A172 Appendix to TNO-report of 3M-02 Final version sample 63 64 63 64 63 64 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 Measured protein conc (mg/ml) 171.8 193 146.9 140.8 166.5 177 147.9 132.7 130.7 127.7 135.7 145.9 131.7 139.7 77.6 72.1 153.1 160.2 132.7 126.7 96.4 90.2 118.4 125.3 162.2 140.6 152.9 177.5 165.9 175.4 129.7 119.4 138.7 113.3 261.1 217.1 176.4 175.4 153.9 138.7 154.8 120.1 167.9 154.9 126.1 125.3 133.2 140.6 169 176.1 174.3 186.3 167.9 155.9 176.4 164.9 150 149 188.5 184.1 189.6 221.4 166.9 161.9 166.9 156.9 230.1 218.4 149 145.2 125.3 132.3 172.2 151.9 170.1 185.2 170.1 176.4 163.9 170.1 132.3 126.1 148.1 130.6 203.6 196.5 212.1 195.4 146.2 213.4 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 average protein conc (mg/ml) 729.6 575.4 687.0 561.2 516.8 563.2 542.8 299.4 626.6 518.8 373.2 487.4 605.6 660.8 682.6 498.2 504.0 956.4 703.6 585.2 549.8 645.6 502.8 547.6 690.2 721.2 647.6 682.6 598.0 745.2 822.0 657.6 647.6 897.0 588.4 515.2 648.2 710.6 693.0 668.0 516.8 557.4 800.2 815.0 719.2 g protein per well 243.2 191.8 229.0 187.1 172.3 187.7 180.9 99.8 208.9 172.9 124.4 162.5 201.9 220.3 227.5 166.1 168.0 318.8 234.5 195.1 183.3 215.2 167.6 182.5 230.1 240.4 215.9 227.5 199.3 248.4 274.0 219.2 215.9 299.0 196.1 171.7 216.1 236.9 231.0 222.7 172.3 185.8 266.7 271.7 239.7 04-12-2006 Page A157 of A172 Page A158 of A172 sample compound concentration 1 control 2 control 3 control 4 control 5 control 6 control 7 control 8 control 9 control 10 control 11 control 12 control 13 PFBS 1 M 14 PFBS 1 M 15 PFBS 10 M 16 PFBS 10 M 17 PFBS 100 M 18 PFBS 100 M 19 PFHS 1 M 20 PFHS 1 M 21 PFHS 10 M 22 PFHS 10 M 23 PFHS 100 M 24 PFHS 100 M 25 PFOS 1 M 26 PFOS 1 M 27 PFOS 10 M 28 PFOS 10 M 29 PFOS 100 M 30 PFOS 100 M 31 Rosuvastatin 0.003 M 32 Rosuvastatin 0.003 M 33 Rosuvastatin 0.03 M 34 Rosuvastatin 0.03 M 35 Rosuvastatin 0.3 M 36 Rosuvastatin 0.3 M 37 PFBS 1 M 38 PFBS 1 M 39 PFBS 10 M 40 PFBS 10 M 41 PFBS 100 M 42 PFBS 100 M 43 PFHS 1 M 44 PFHS 1 M 45 PFHS 10 M 46 PFHS 10 M 47 PFHS 100 M 48 PFHS 100 M 49 PFOS 1 M 50 PFOS 1 M 51 PFOS 10 M 52 PFOS 10 M 53 PFOS 100 M 54 PFOS 100 M 14C Label 14C acetic acid 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 3H-cholesterol 14C-cholesterol 14C-cholesterol corrected g protein 14C-cholesterol Average14C-cholesterol % of control Average SD (dpm) (dpm) for recovery (dpm) per mg protein (dpm/mg) per mg protein (dpm/mg) % of control 12705 7957 16067 125.4 128130 129029 99 100 0.7 12827 10612 21225 163.3 129974 101 11820 12414 26944 208.9 128982 100 14124 8971 16295 224.1 72713 108158 67 100 34.2 12699 11979 24200 229.9 105265 97 15542 14954 24684 168.5 146495 135 12531 17327 35474 169.5 209286 156534 134 100 31.0 11864 16561 35812 314.8 113761 73 11644 15565 34294 234 146556 94 14154 7968 14442 198.3 72832 70783 103 100 19.5 12949 7804 15462 185.3 83441 118 14391 6763 12056 215 56077 79 15327 11500 19249 170.8 112700 110093 87 85 2.9 14468 11220 19896 185.1 107486 83 14439 13928 24747 231.9 106714 105182 83 82 1.7 18148 17531 24783 239.1 103650 80 12484 10821 22237 216.2 102856 97594 80 76 5.8 13353 10981 21098 228.5 92331 72 15401 13298 22152 201.9 109717 117064 85 91 8.1 14081 16955 30891 248.3 124411 96 14983 15087 25833 280 92261 105305 72 82 14.3 13978 14244 26143 220.9 118349 92 14523 14505 25623 216.9 118134 107898 92 84 11.2 16419 18682 29191 298.9 97661 76 13313 9191 17712 198.1 89408 120763 69 94 34.4 13999 14418 26423 173.7 152118 118 13096 11960 23430 216.5 108220 103774 84 80 4.9 14417 13268 23610 237.7 99328 77 12438 8472 17475 232.6 75127 83236 58 65 8.9 12519 9940 20370 223.0 91345 71 13661 6222 11685 174.3 67038 58119 52 45 9.8 14096 5074 9235 187.7 49200 38 12455 1670 3440 265.6 12951 14780 10 11 2.0 12791 2268 4549 273.9 16608 13 14394 624 1112 261.6 4251 4151 3 3 0.1 13181 465 905 223.4 4051 3 13006 7667 15124 273.9 55216 66147 51 61 14.3 13189 10370 20172 261.7 77079 71 13331 17500 33678 223.5 150685 138237 139 128 16.3 12774 14299 28718 228.3 125790 116 13454 8093 15432 189.7 81351 100007 75 92 24.4 16489 14468 22511 189.7 118664 110 12397 13939 28846 202.0 142802 124204 132 115 24.3 12966 11630 23012 217.9 105606 98 13703 10974 20546 216.5 94899 109301 88 101 18.8 13864 15549 28773 232.6 123702 114 15672 16480 26978 244.0 110564 108003 102 100 3.3 15719 17411 28417 269.5 105442 97 12626 9783 19878 282.5 70366 82685 65 76 16.1 13507 16521 31380 330.3 95004 88 15475 19586 32470 327.3 99207 105912 92 98 8.8 13164 12135 23650 210.0 112617 104 15165 9834 16636 264.5 62898 62847 58 58 0.1 15228 10776 18155 289.1 62797 58 [14C]-Cholesterol synthesis in rat hepatocytes 04-12-2006 Final version Appendix to TNO-report of 3M-02 Page A159 of A172 sample 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 compound concentration Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M 14C Label 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C 14C input input input palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid palmitic acid 3H cholesterol 3H cholesterol 3H cholesterol average input 3H cholesterol 3H-cholesterol (dpm) 15609 15687 14378 13730 15870 12832 13019 16614 16141 15873 17295 16841 10210 15012 15875 16131 16397 14452 15496 17004 13663 13951 14919 16126 15126 15117 15845 15353 16583 15158 14410 15961 16275 14927 14899 2959 14864 15988 14366 15716 13749 16009 14012 15284 14474 12042 14571 13686 13478 13285 12506 13476 14963 26014 25923 25028 25655 14C-cholesterol (dpm) 4123 4793 901 984 311 512 11827 19187 21002 19906 21050 16813 11716 18465 17804 20213 12748 11356 14299 17756 13984 16405 11557 12122 4914 5462 1201 1756 465 505 5450 5718 5596 5781 5367 1505 6675 4773 5602 8024 5977 5872 4048 9125 5125 5104 4478 4542 3476 2874 1940 1859 1145 14C-cholesterol corrected for recovery (dpm) 6777 7839 1608 1839 503 1024 23306 29628 33381 32173 31225 25612 29439 31556 28772 32147 19946 20159 23673 26790 26258 30168 19874 19285 8335 9270 1945 2934 719 855 9703 9191 8821 9936 9242 13049 11521 7659 10004 13098 11153 9410 7412 15317 9084 10874 7884 8514 6616 5550 3980 3539 1963 g protein 268.9 303.1 276.0 308.9 283.3 253.1 175.6 224.9 243.2 191.8 229.0 187.1 172.3 187.7 180.9 99.8 208.9 172.9 124.4 162.5 201.9 220.3 227.5 166.1 168.0 318.8 234.5 195.1 183.3 215.2 167.6 182.5 230.1 240.4 215.9 228.5 199.3 248.4 274.0 219.2 215.9 299.0 196.1 171.7 216.1 236.9 231.0 222.7 172.3 185.8 266.7 271.7 239.7 14C-cholesterol per mg protein (dpm/mg) 25201 25861 5825 5952 1775 4044 132722 131739 137258 167745 136354 136891 170860 168120 159051 322115 95480 116594 190300 164859 130053 136939 87357 116105 49611 29076 8292 15040 3925 3972 57894 50361 38336 41330 42805 57105 57807 30833 36511 59756 51657 31472 37795 89207 42036 45901 34131 38232 38401 29871 14922 13026 8190 Average14C-cholesterol per mg protein (dpm/mg) 25531 5889 2910 132231 152501 136623 168120 240583 106037 177579 133496 101731 39343 11666 3948 54127 39833 42805 44320 48134 41565 63501 43968 36182 34136 13974 8190 % of control Average % of control 23 24 24 5 5 6 2 3 4 85 84 84 88 97 107 87 87 87 109 108 107 102 154 206 61 68 74 122 113 105 83 85 87 56 65 74 32 25 19 5 7 10 3 3 3 82 76 71 54 56 58 60 71 81 82 63 44 52 68 84 73 59 44 53 90 126 59 62 65 48 51 54 54 48 42 21 20 18 12 12 04-12-2006 Final version Appendix to TNO-report of 3M-02 Appendix to TNO-report of 3M-02 Experiment 2 Final version 04-12-2006 Protein concentration determination sample 1 2 3 4 5 6 7 8 9 10 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 Measured protein conc (mg/ml) 191.5 204.6 218.1 230.3 234.8 221 267.6 215.2 187.7 214.2 222.9 233.8 235.8 266.6 250 260.3 232.8 241.9 246.9 269.7 211.4 195.2 155.9 154.1 123.5 172.1 156.8 231.8 139.1 114.9 147 194.3 256.2 220.1 126.6 143.5 221 217.1 158.6 179.4 235.8 224.9 175.8 241.8 196.3 204.6 206.6 200.8 240.8 242.9 190.5 191.5 150.6 154.1 142.6 140.9 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 average protein conc (mg/ml) 792.2 896.8 911.6 965.6 803.8 913.4 1004.8 1020.6 949.4 1033.2 813.2 620.0 591.2 777.2 508.0 682.6 952.6 540.2 876.2 676.0 921.4 835.2 801.8 814.8 967.4 764.0 609.4 567.0 g protein per well 264.1 298.9 303.9 321.9 267.9 304.5 334.9 340.2 316.5 344.4 271.1 206.7 197.1 259.1 169.3 227.5 317.5 180.1 292.1 225.3 307.1 278.4 267.3 271.6 322.5 254.7 203.1 189.0 sample 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 Measured protein conc (mg/ml) 231.9 240.9 211.3 208.5 216.2 239.8 203.7 235.8 292.4 340.6 167.6 197.1 177.5 211.3 237.4 232.4 217.3 244.9 202.1 206.2 240 266.7 181.8 202.9 217.3 223.1 188.5 174.1 238.2 250 173.3 204.6 235.8 231.6 229.9 240 183.5 188.5 163.1 152 202.9 228.1 213.8 244.9 211.4 218.1 168.2 149.4 211.4 213 232.4 254.2 245.8 224 156.2 136.5 215.6 236.6 133.9 158.7 210.5 209.7 156.2 184.3 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 average protein conc (mg/ml) 945.6 839.6 912.0 879.0 1266.0 729.4 777.6 939.6 924.4 816.6 1013.4 769.4 880.8 725.2 976.4 755.8 934.8 939.8 744.0 630.2 862.0 917.4 859.0 635.2 848.8 973.2 939.6 585.4 904.4 585.2 840.4 681.0 g protein per well 315.2 279.9 304.0 293.0 422.0 243.1 259.2 313.2 308.1 272.2 337.8 256.5 293.6 241.7 325.5 251.9 311.6 313.3 248.0 210.1 287.3 305.8 286.3 211.7 282.9 324.4 313.2 195.1 301.5 195.1 280.1 227.0 Page A160 of A172 Appendix to TNO-report of 3M-02 Final version sample 63 64 63 64 63 64 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 Measured protein conc (mg/ml) 252.5 225.7 153.6 142.5 228.2 210.5 228.1 204.6 220.6 244.2 133.9 150.2 220.6 224.8 234.9 236.6 247.5 230.7 137.3 156.4 206.2 194.7 150 175 152.7 142.8 141.9 140.1 230.3 233.8 237.3 211.6 184 146.9 118.4 200.1 180.4 235.6 162.6 132 211.6 252.4 168.8 170.6 222.3 242.6 257.7 247.1 223.1 208.9 178.7 215.1 179.6 194.7 168.8 142.9 188.5 222.3 196.5 180.4 226.7 240.9 182.2 228.4 240 165.2 142.8 272.7 243.5 201.8 167.1 213.4 236.5 254.2 219.6 153.6 122 232 281.5 207.2 227.6 208.1 207.2 192.9 161.7 281.6 218.7 145.5 151.8 162.6 137.4 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 average protein conc (mg/ml) 252 256 252 256 252 256 276 280 284 288 292 296 300 304 308 312 316 320 324 328 332 336 340 344 348 352 356 360 364 368 372 376 380 384 388 392 396 400 404 408 412 416 420 424 428 432 g protein per well 84.0 85.3 84.0 85.3 84.0 85.3 92.0 93.3 94.7 96.0 97.3 98.7 100.0 101.3 102.7 104.0 105.3 106.7 108.0 109.3 110.7 112.0 113.3 114.7 116.0 117.3 118.7 120.0 121.3 122.7 124.0 125.3 126.7 128.0 129.3 130.7 132.0 133.3 134.7 136.0 137.3 138.7 140.0 141.3 142.7 144.0 04-12-2006 Page A161 of A172 Page A162 of A172 sample compound concentration 1 control 2 control 3 control 4 control 5 control 6 control 7 control 8 control 9 control 13 PFBS 1 M 14 PFBS 1 M 15 PFBS 10 M 16 PFBS 10 M 17 PFBS 100 M 18 PFBS 100 M 19 PFHS 1 M 20 PFHS 1 M 21 PFHS 10 M 22 PFHS 10 M 23 PFHS 100 M 24 PFHS 100 M 25 PFOS 1 M 26 PFOS 1 M 27 PFOS 10 M 28 PFOS 10 M 29 PFOS 100 M 30 PFOS 100 M 31 Rosuvastatin 0.003 M 32 Rosuvastatin 0.003 M 33 Rosuvastatin 0.03 M 34 Rosuvastatin 0.03 M 35 Rosuvastatin 0.3 M 36 Rosuvastatin 0.3 M 37 PFBS 1 M 38 PFBS 1 M 39 PFBS 10 M 40 PFBS 10 M 41 PFBS 100 M 42 PFBS 100 M 43 PFHS 1 M 44 PFHS 1 M 45 PFHS 10 M 46 PFHS 10 M 47 PFHS 100 M 48 PFHS 100 M 49 PFOS 1 M 50 PFOS 1 M 51 PFOS 10 M 52 PFOS 10 M 53 PFOS 100 M 54 PFOS 100 M 14C Label 14C acetic acid 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 3H-cholesterol (dpm) 10176 15067 8013 7102 13239 16182 11285 9341 14585 10415 11106 13209 17775 11729 16244 15747 13331 15569 8199 15486 14084 15835 16929 15411 9454 15895 17054 18199 17403 16235 14014 15647 9439 17104 16962 16583 15691 14615 16380 16700 15447 15347 15658 16797 16476 14120 13621 16279 16347 15609 16385 14C-cholesterol (dpm) 13013 29921 20565 3628 7313 13946 19393 14887 19635 15531 22304 31195 41007 24081 42002 34491 30717 28923 21682 25911 36299 30330 46461 37008 20098 26782 29629 16110 11966 3127 1836 499 439 8190 23749 11608 16193 17212 17173 12743 20907 18404 22910 19524 25363 11804 21880 28289 17820 18990 28115 14C-cholesterol corrected for recovery (dpm) 34516 53601 69272 13788 14910 23262 46384 43017 36337 40250 54206 63744 62269 55416 69791 59120 62193 50143 71378 45162 69565 51699 74077 64817 57380 45479 46894 23893 18559 5199 3536 861 1255 12924 37791 18894 27855 31788 28298 20596 36532 32368 39492 31373 41550 24616 43357 46904 29423 32838 46314 g protein 264.1 298.9 303.9 321.9 267.9 304.5 334.9 340.2 316.5 271.1 206.7 197.1 259.1 169.3 227.5 317.5 180.1 292.1 225.3 307.1 278.4 267.3 271.6 322.5 254.7 203.1 189.0 315.2 279.9 304.0 293.0 422.0 243.1 259.2 313.2 308.1 272.2 337.8 256.5 293.6 241.7 325.5 251.9 311.6 313.3 248.0 210.1 287.3 305.8 286.3 211.7 14C-cholesterol per mg protein (dpm/mg) 130711 179328 227969 42839 55647 76402 138487 126446 114821 148487 262288 323464 240359 327262 306730 186184 345324 171682 316765 147042 249876 193434 272742 201004 225315 223922 248115 75803 66313 17101 12069 2040 5163 49863 120662 61317 102332 94102 110338 70149 151125 99450 156757 100685 132635 99258 206398 163241 96218 114684 218739 Average14C-cholesterol per mg protein (dpm/mg) 179336 58296 126585 205388 281912 316996 265754 244224 198459 233088 213159 236019 71058 14585 3601 85262 81825 102220 110637 128104 116660 152828 129729 166712 % of control Average % of control 73 100 100 127 73 100 95 131 83 115 146 180 157 134 182 177 171 104 148 193 96 136 177 82 111 139 108 130 152 112 119 126 125 132 138 42 40 37 10 8 7 1 2 3 86 146 207 105 140 176 161 175 189 120 190 259 171 220 269 173 200 228 170 262 354 280 223 165 197 286 375 SD 27.1 29.1 44.9 32.8 8.1 62.7 57.2 40.5 31.3 9.6 9.5 3.7 2.0 1.2 85.9 49.8 19.7 98.2 69.5 38.8 130.0 81.3 126.2 [14C]-Cholesterol synthesis in rat hepatocytes 04-12-2006 Final version Appendix to TNO-report of 3M-02 sample compound concentration 14C Label 3H-cholesterol 14C-cholesterol 14C-cholesterol corrected g protein 14C-cholesterol Average14C-cholesterol % of control Average SD (dpm) (dpm) for recovery (dpm) per mg protein (dpm/mg) per mg protein (dpm/mg) % of control Appendix to TNO-report of 3M-02 55 Rosuvastatin 0.003 M 14C butyric acid 16224 4328 7200 282.9 25449 27019 44 46 3.8 56 Rosuvastatin 0.003 M 14C butyric acid 16103 5533 9274 324.4 28589 49 57 Rosuvastatin 0.03 M 58 Rosuvastatin 0.03 M 14C butyric acid 14C butyric acid 16808 17593 975 1781 1566 2732 313.2 195.1 4999 14003 9501 9 16 10.9 24 59 Rosuvastatin 0.3 M 14C butyric acid 14906 452 818 301.5 2715 2604 5 4 0.3 60 Rosuvastatin 0.3 M 14C butyric acid 15758 284 486 195.1 2494 4 61 PFBS 1 M 14C lauric acid 14491 19019 38645 280.1 137952 165768 109 131 31.1 62 PFBS 1 M 14C lauric acid 17266 28110 43943 227.0 193583 153 63 PFBS 10 M 14C lauric acid 16567 21487 35007 318.8 109809 129179 87 102 21.6 64 PFBS 10 M 14C lauric acid 17868 19412 29324 197.4 148550 117 65 PFBS 100 M 14C lauric acid 17520 21648 33351 292.5 114033 127014 90 100 14.5 66 PFBS 100 M 14C lauric acid 19364 28972 40384 288.5 139995 111 67 PFHS 1 M 14C lauric acid 16638 26282 42637 309.9 137582 168120 109 150 58.5 68 PFHS 1 M 14C lauric acid 17741 30159 45884 189.4 242261 191 69 PFHS 10 M 14C lauric acid 17115 25762 40628 296.9 136826 130644 108 103 6.9 70 PFHS 10 M 14C lauric acid 16449 23842 39123 314.3 124462 98 71 PFHS 100 M 14C lauric acid 15800 21356 36483 318.8 114438 169881 90 134 61.9 72 PFHS 100 M 14C lauric acid 16793 27449 44119 195.8 225325 178 73 PFOS 1 M 14C lauric acid 16050 18811 31635 267.3 118363 150205 94 119 35.6 74 PFOS 1 M 14C lauric acid 17035 24894 39444 216.7 182048 144 75 PFOS 10 M 14C lauric acid 16660 16687 27035 197.0 137234 157048 108 124 22.1 76 PFOS 10 M 14C lauric acid 15663 19295 33250 188.0 176863 140 77 PFOS 100 M 14C lauric acid 16046 27259 45853 309.4 148200 157868 117 125 10.8 78 PFOS 100 M 14C lauric acid 13213 24544 50138 299.3 167537 132 Final version 79 Rosuvastatin 0.003 M 14C lauric acid 13776 6081 11915 220.6 54010 44273 43 35 10.9 80 Rosuvastatin 0.003 M 14C lauric acid 12989 3529 7333 212.3 34537 27 81 Rosuvastatin 0.03 M 82 Rosuvastatin 0.03 M 14C lauric acid 14C lauric acid 15098 11184 1767 2114 3159 5102 277.3 196.4 11390 25977 18684 9 15 8.1 21 83 Rosuvastatin 0.3 M 14C lauric acid 13447 566 1136 309.3 3673 3183 3 3 0.5 84 Rosuvastatin 0.3 M 14C lauric acid 14569 329 610 226.3 2694 2 input 3H cholesterol 26701 input 3H cholesterol input 3H cholesterol 27378 26895 average input 3H cholesterol 26991.33333 Page A163 of A172 04-12-2006 Appendix to TNO-report of 3M-02 Experiment 3 Final version Protein concentration determination sample 1 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 Measured protein conc (mg/ml) 246.35 298.46 229.35 266.32 232.70 299.74 186.54 154.56 143.00 169.33 153.58 301.02 220.50 370.42 222.70 224.91 171.32 272.34 180.40 240.62 227.12 209.66 295.91 248.65 295.91 331.42 268.72 222.71 282.13 236.08 172.33 263.93 175.34 171.32 247.50 249.81 393.62 348.03 289.60 153.58 227.12 209.66 317.98 272.34 211.81 258.00 195.88 304.88 389.03 352.27 258.00 279.67 363.75 295.91 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 protein conc (mg/ml) 985.4 1193.8 917.4 1065.3 930.8 1199.0 746.2 618.2 572.0 677.3 614.3 1204.1 882.0 1481.7 890.8 899.6 685.3 1089.4 721.6 962.5 908.5 838.6 1183.6 994.6 1183.6 1325.7 1074.9 890.8 1128.5 944.3 689.3 1055.7 701.4 685.3 990.0 999.2 1574.5 1392.1 1158.4 614.3 908.5 838.6 1271.9 1089.4 847.2 1032.0 783.5 1219.5 1556.1 1409.1 1032.0 1118.7 1455.0 1183.6 g protein per well 328.5 397.9 305.8 355.1 310.3 399.7 248.7 206.1 190.7 225.8 204.8 401.4 294.0 493.9 296.9 299.9 228.4 363.1 240.5 320.8 302.8 279.5 394.5 331.5 394.5 441.9 358.3 296.9 376.2 314.8 229.8 351.9 233.8 228.4 330.0 333.1 524.8 464.0 386.1 204.8 302.8 279.5 424.0 363.1 282.4 344.0 261.2 406.5 518.7 469.7 344.0 372.9 485.0 394.5 04-12-2006 Page A164 of A172 Appendix to TNO-report of 3M-02 Final version sample 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 Measured protein conc (mg/ml) 356.51 319.45 356.51 319.45 226.01 241.05 333.17 251.23 297.09 232.80 268.15 294.15 222.82 295.13 244.74 237.38 207.61 324.00 253.09 241.05 283.46 267.20 275.77 347.64 216.53 368.75 219.00 329.09 221.92 285.39 348.30 312.93 212.95 288.30 281.53 305.97 245.66 274.81 289.28 254.03 230.98 288.30 332.15 294.15 219.22 335.22 252.16 297.09 247.52 280.57 250.30 296.11 217.43 dilution 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 protein conc (mg/ml) 1426.0 1277.8 1426.0 1277.8 904.0 964.2 1332.7 1004.9 1188.4 931.2 1072.6 1176.6 891.3 1180.5 979.0 949.5 830.4 1296.0 1012.4 964.2 1133.8 1068.8 1103.1 1390.6 866.1 1475.0 876.0 1316.4 887.7 1141.6 1393.2 1251.7 851.8 1153.2 1126.1 1223.9 982.6 1099.2 1157.1 1016.1 923.9 1153.2 1328.6 1176.6 876.9 1340.9 1008.6 1188.4 990.1 1122.3 1001.2 1184.4 869.7 g protein per well 475.3 425.9 475.3 425.9 301.3 321.4 444.2 335.0 396.1 310.4 357.5 392.2 297.1 393.5 326.3 316.5 276.8 432.0 337.5 321.4 377.9 356.3 367.7 463.5 288.7 491.7 292.0 438.8 295.9 380.5 464.4 417.2 283.9 384.4 375.4 408.0 327.5 366.4 385.7 338.7 308.0 384.4 442.9 392.2 292.3 447.0 336.2 396.1 330.0 374.1 333.7 394.8 289.9 04-12-2006 Page A165 of A172 Page A166 of A172 3H cholesterol mix 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 sample 1 3 4 5 6 7 8 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 compound concentration control control control control control control control control control PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M PFBS 1 M PFBS 1 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M 14C Label 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C lauric acid 14C lauric acid 14C palmitic acid 14C palmitic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 3H-cholesterol (dpm) 12507 19023 16324 15491 14301 15647 14301 16834 17733 17724 15643 13898 17481 17149 18303 17772 16907 17592 16152 18689 17098 17841 19413 17762 14828 16243 15814 15332 14969 14770 14215 16761 16727 15968 14142 16118 16293 19457 18922 17479 17443 16118 18498 17002 17226 16924 15074 17062 15031 18507 14681 15247 15905 17593 13507 18268 14C-cholesterol (dpm) 44451 99078 18935 15714 17937 40817 32511 5000 4789 22246 21849 22496 26287 23557 22881 35195 39844 23683 24871 27769 22118 27624 43559 25533 24114 22967 26323 2953 3029 574 626 257 248 5934 11778 13512 9931 7769 6439 14976 7993 9922 8585 10438 7974 8198 12993 18014 12106 9530 930 687 276 212 94 120 14C-cholesterol corrected for recovery (dpm) 91111 133519 29736 26005 32153 66874 58279 7614 6923 32176 35806 41495 38550 35215 32048 50768 60414 34512 39474 38091 33162 39693 57521 36851 41690 36248 42672 4938 5187 996 1129 393 380 9527 21350 21491 15626 10236 8724 21965 11747 15781 11898 15738 11867 12418 22097 27066 20647 13201 1624 1155 445 309 178 168 g protein 328.5 397.9 305.8 355.1 310.3 399.7 248.7 225.8 204.8 401.4 294.0 493.9 296.9 299.9 228.4 363.1 240.5 320.8 302.8 279.5 394.5 331.5 394.5 441.9 358.3 296.9 376.2 314.8 229.8 351.9 233.8 228.4 330.0 333.1 524.8 464.0 204.8 302.8 279.5 424.0 363.1 282.4 344.0 261.2 406.5 518.7 469.7 344.0 372.9 485.0 394.5 475.3 425.9 475.3 425.9 301.3 14C-cholesterol per mg protein (dpm/mg) 277384 335520 97240 73233 103632 167329 234314 33725 33809 80168 121789 84018 129826 117430 140298 139810 251169 107571 130352 136259 84052 119725 145791 83394 116357 122069 113436 15686 22576 2831 4829 1721 1152 28602 40681 46312 76307 33802 31206 51807 32351 55879 34586 60261 29192 23940 47045 78680 55370 27218 4116 2430 1044 650 419 559 Average14C-cholesterol per mg protein (dpm/mg) 306452 91368 200821 33767 100979 106922 128864 195490 118962 110155 132758 99876 117752 19131 3830 1436 34641 46312 55054 41506 44115 47423 26566 62863 41294 3273 847 489 % of control Average SD % of control 90.5 100.0 13.4 109.5 106.4 100.0 17.5 80.2 113.4 83.3 100.0 23.6 116.7 99.9 100.0 0.18 100.1 26.2 33.0 9.6 39.7 27.4 34.9 10.6 42.4 38.3 42.1 5.3 45.8 45.6 63.8 25.7 82.0 35.1 38.8 5.3 42.5 44.5 35.9 12.0 27.4 39.1 43.3 6.0 47.6 27.2 32.6 7.6 38.0 39.8 38.4 2.0 37.0 5.1 6.2 1.6 7.4 0.9 1.2 0.5 1.6 0.6 0.5 0.1 0.4 31.3 37.9 9.3 44.5 50.7 50.7 83.5 60.3 32.9 37.0 34.2 45.4 15.9 56.7 35.4 48.3 18.2 61.2 37.9 51.9 19.9 66.0 32.0 29.1 4.1 26.2 51.5 68.8 24.5 86.1 60.6 45.2 21.8 29.8 4.5 3.6 1.3 2.7 1.1 0.9 0.3 0.7 0.5 0.5 0.1 0.6 [14C]-Cholesterol synthesis in rat hepatocytes 04-12-2006 Final version Appendix to TNO-report of 3M-02 Page A167 of A172 3H cholesterol mix 1 1 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 sample 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 compound concentration PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M 14C Label 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C lauric acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid input 3H cholesterol input 3H cholesterol input 3H cholesterol average input 3H cholesterol 3H-cholesterol (dpm) 18064 16312 16632 16430 17140 16929 17947 17680 17511 18652 17842 17390 12194 17125 16554 16019 15880 17209 15981 14109 17448 16191 15558 16555 17040 15827 16659 15285 17337 14572 14708 15562 14849 16477 15761 16923 19055 16210 17729 16090 17458 12554 17205 16635 17350 13361 15847 mix 1 25860 25671 25376 25635.7 14C-cholesterol (dpm) 35197 33843 31024 42519 24245 22983 34462 35488 31561 34500 20913 30612 39952 49945 29563 42312 22742 31390 3218 3171 754 686 261 4312 3219 3366 4602 2953 3284 2399 3322 3238 3857 2952 3697 3859 4608 4074 5568 2981 3072 711 986 534 544 313 359 mix 2 24418 25195 24950 24854.3 14C-cholesterol corrected for recovery (dpm) 49950 53187 47819 66342 36262 34803 49226 51457 46205 47417 30048 45127 81432 72488 44386 65649 35594 45335 5005 5586 1074 1053 417 6474 4695 5286 6866 4802 4708 4092 5614 5171 6456 4453 5830 5668 6010 6247 7806 4605 4373 1408 1424 798 779 582 563 g protein 321.4 444.2 335.0 396.1 310.4 357.5 392.2 297.1 393.5 326.3 316.5 276.8 432.0 337.5 321.4 377.9 356.3 367.7 463.5 288.7 491.7 292.0 438.8 380.5 464.4 417.2 283.9 384.4 375.4 408.0 327.5 366.4 385.7 338.7 308.0 384.4 442.9 392.2 292.3 447.0 336.2 396.1 330.0 374.1 333.7 394.8 289.9 14C-cholesterol per mg protein (dpm/mg) 155414 119730 142754 167480 116825 97343 125512 173201 117417 145310 94937 163023 188500 214808 138103 173700 99909 123297 10797 19348 2185 3606 950 17013 10110 12669 24182 12492 12542 10030 17139 14114 16738 13147 18930 14744 13572 15927 26705 10302 13008 3554 4316 2133 2335 1475 1942 Average14C-cholesterol per mg protein (dpm/mg) 137572 155117 107084 168120 131364 128980 201654 155901 111603 15073 2895 1050 13562 18425 42805 13584 15426 16038 14158 21316 11655 3935 2234 8190 % of control Average SD % of control 77.4 68.5 12.6 59.6 71.1 77.2 8.7 83.4 58.2 53.3 6.9 48.5 62.5 74.4 16.8 86.2 58.5 65.4 9.8 72.4 47.3 64.2 24.0 81.2 93.9 100.4 9.3 107.0 68.8 77.6 12.5 86.5 49.8 55.6 8.2 61.4 5.4 7.5 3.0 9.6 1.1 1.4 0.5 1.8 0.5 0.5 0.1 50.4 50.4 37.5 54.6 24.1 71.6 37.0 37.1 0.1 37.1 29.7 40.2 14.9 50.8 41.8 45.7 5.5 49.6 38.9 47.5 12.1 56.1 43.7 41.9 2.5 40.2 47.2 63.1 22.6 79.1 30.5 34.5 5.7 38.5 10.5 11.7 1.6 12.8 6.3 6.6 0.4 6.9 4.4 5.1 1.0 5.8 04-12-2006 Final version Appendix to TNO-report of 3M-02 Appendix to TNO-report of 3M-02 Experiment 4 Final version 04-12-2006 Protein concentration determination sample 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 Measured protein conc (mg/ml) well 1 well 2 207.81 223.77 214.31 224.72 189.73 202.31 205.97 230.5 234.4 252.34 221.86 205.97 218.07 224.72 225.68 225.68 214.31 223.77 210.58 214.31 210.58 220.91 199.58 205.97 212.44 214.31 241.29 245.28 230.5 226.54 234.4 239.31 272.05 262.61 292.63 272.24 254.37 242.28 231.47 237.34 269.94 274.18 272.05 253.35 253.35 250.31 242.28 221.86 202.31 210.58 241.29 242.28 245.28 239.31 230.5 235.37 245.28 233.42 218.07 209.66 250.31 240.3 253.35 242.28 244.28 249.3 233.42 237.34 215.24 204.14 227.6 210.58 249.52 228.83 256.56 246.52 277.1 284.44 248.52 238.6 251.52 254.54 266.75 258.59 238.6 246.52 265.73 254.54 251.52 253.53 242.55 245.53 237.62 229.8 212.56 208.8 211.62 203.19 206.92 209.73 194.87 192.13 206.92 210.68 222.08 234.67 177.7 186.68 214.45 228.83 220.17 214.45 dilution 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 protein conc (mg/ml) 1079.0 1097.6 980.1 1091.2 1216.9 1069.6 1107.0 1128.4 1095.2 1062.2 1078.7 1013.9 1066.9 1216.4 1142.6 1184.3 1336.7 1412.2 1241.6 1172.0 1360.3 1313.5 1259.2 1160.4 1032.2 1208.9 1211.5 1164.7 1196.8 1069.3 1226.5 1239.1 1234.0 1176.9 1048.5 1095.5 1195.9 1257.7 1403.9 1217.8 1265.2 1313.4 1212.8 1300.7 1262.6 1220.2 1168.6 1053.4 1037.0 1041.6 967.5 1044.0 1141.9 911.0 1108.2 1086.6 g protein per well 359.7 365.9 326.7 363.7 405.6 356.5 369.0 376.1 365.1 354.1 359.6 338.0 355.6 405.5 380.9 394.8 445.6 470.7 413.9 390.7 453.4 437.8 419.7 386.8 344.1 403.0 403.8 388.2 398.9 356.4 408.8 413.0 411.3 392.3 349.5 365.2 398.6 419.2 468.0 405.9 421.7 437.8 404.3 433.6 420.9 406.7 389.5 351.1 345.7 347.2 322.5 348.0 380.6 303.7 369.4 362.2 Page A168 of A172 Appendix to TNO-report of 3M-02 Final version 04-12-2006 sample 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 91 92 94 95 96 97 98 100 101 103 104 105 106 107 109 110 112 113 114 115 116 118 119 Measured protein conc (mg/ml) well 1 well 2 224.96 223.04 219.21 218.26 213.51 204.12 199.48 205.99 210.68 204.12 186.68 193.04 160.17 161.9 209.73 203.19 205.99 217.3 229.8 225.93 237.62 246.52 222.08 234.67 261.64 263.68 259.6 235.65 239.59 255.55 278.14 285.49 212.61 215.22 232.92 229.35 225.79 240.13 231.13 233.82 233.82 241.03 225.79 239.22 241.03 238.32 239.22 232.03 217.85 221.37 224.02 234.72 220.49 225.79 283 287.81 208.27 190.32 204.81 214.35 203.09 217.85 218.73 214.35 216.10 212.61 205.67 203.95 224.02 217.85 218.73 211.74 233.82 224.02 226.68 218.73 242.85 235.62 252.91 239.22 242.85 245.58 218.73 209.13 255.68 256.60 242.85 245.58 248.32 237.42 229.35 242.85 251.99 243.76 245.58 259.39 231.13 217.85 251.07 255.68 272.51 291.69 219.61 216.10 243.74 246.67 255.52 269.48 267.47 271.50 234.05 235.01 dilution 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 protein conc (mg/ml) 1120.0 1093.7 1044.1 1013.7 1037.0 949.3 805.2 1032.3 1058.2 1139.3 1210.4 1141.9 1313.3 1238.1 1237.9 1409.1 1069.6 1155.7 1164.8 1162.4 1187.1 1162.5 1198.4 1178.1 1098.1 1146.9 1115.7 1427.0 996.5 1047.9 1052.4 1082.7 1071.8 1024.1 1104.7 1076.2 1144.6 1113.5 1196.2 1230.3 1221.1 1069.7 1280.7 1221.1 1214.4 1180.5 1239.4 1262.4 1122.5 1266.9 1410.5 1089.3 1226.0 1312.5 1347.4 1172.7 g protein per well 373.3 364.6 348.0 337.9 345.7 316.4 268.4 344.1 352.7 379.8 403.5 380.6 437.8 412.7 412.6 469.7 356.5 385.2 388.3 387.5 395.7 387.5 399.5 392.7 366.0 382.3 371.9 475.7 332.2 349.3 350.8 360.9 357.3 341.4 368.2 358.7 381.5 371.2 398.7 410.1 407.0 356.6 426.9 407.0 404.8 393.5 413.1 420.8 374.2 422.3 470.2 363.1 408.7 437.5 449.1 390.9 Page A169 of A172 Page A170 of A172 3H cholesterol mix 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 sample 1 2 3 4 5 7 8 9 10 11 12 13 15 17 18 19 20 22 23 24 25 26 27 28 30 31 32 33 34 35 36 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 56 57 58 compound concentration control control control control control control control control control control control PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M PFBS 1 M PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M 14C Label 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C palmitic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C acetic acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 14C butyric acid 3H-cholesterol 14C-cholesterol 14C-cholesterol corrected g protein 14C-cholesterol % of control Average SD (dpm) (dpm) for recovery (dpm) per mg protein (dpm/mg) % of control 11557 1815 3220 359.7 8953 98.5 100.0 12.7 13921 1877 2765 365.9 7556 83.2 12789 2094 3357 326.7 10276 113.1 14330 2430 3477 363.7 9559 105.2 14143 2257 3272 405.6 8067 64.9 100.0 33.3 12409 2888 4772 369.0 12932 104.0 13395 4007 6134 376.1 16307 131.1 12206 1142 1918 365.1 5255 76.4 100.0 16.2 13316 1694 2608 354.1 7367 107.2 12616 1574 2558 359.6 7114 103.5 13307 1703 2624 338.0 7764 112.9 14019 2569 3757 355.6 10566 116.3 90.0 37.2 14018 1506 2203 380.9 5784 63.7 13123 1714 2678 445.6 6011 66.2 67.3 1.6 12859 1837 2929 470.7 6223 68.5 12350 1427 2369 413.9 5724 63.0 93.7 43.4 12583 2709 4414 390.7 11299 124.4 12002 1811 3094 437.8 7066 77.8 91.8 21.4 12074 1822 3094 419.7 7372 81.1 10469 2089 4091 386.8 10578 116.4 12178 2935 4942 344.1 14362 158.1 154.3 20.2 13070 3091 4849 403.0 12033 132.4 11021 3398 6322 403.8 15655 172.3 12494 2059 3379 388.2 8704 95.8 100.0 5.9 11696 1924 3373 356.4 9463 104.1 12079 3650 6196 408.8 15155 166.8 139.9 24.2 12015 2929 4998 413.0 12102 133.2 12687 2772 4480 411.3 10892 119.9 12477 2383 3916 392.3 9982 109.9 113.9 8.6 12560 2407 3929 349.5 11243 123.7 10462 1828 3583 365.2 9811 108.0 10416 1647 3242 419.2 7733 85.1 89.0 5.4 9390 1807 3946 468.0 8432 92.8 9534 81 171 405.9 420 4.6 3.8 1.1 11731 95 163 421.7 386 4.2 12187 61 100 437.8 230 2.5 11494 36 63 404.3 156 1.7 1.4 0.5 12998 42 65 433.6 150 1.6 10852 18 33 420.9 79 0.9 9480 7 15 406.7 36 0.4 0.6 0.1 8849 10 23 389.5 58 0.6 11450 12 21 351.1 60 0.7 9541 1216 2559 345.7 7402 59.5 92.0 36.8 11879 2158 3647 347.2 10504 84.5 12114 3192 5290 322.5 16403 131.9 11350 998 1765 348.0 5073 40.8 52.2 10.1 11456 1499 2627 380.6 6902 55.5 9935 1125 2273 303.7 7487 60.2 11036 4223 7682 362.2 21211 170.6 118.7 73.4 9818 1517 3102 373.3 8309 66.8 12175 1686 2780 364.6 7626 61.3 124.5 62.2 [14C]-Cholesterol synthesis in rat hepatocytes 04-12-2006 Final version Appendix to TNO-report of 3M-02 Page A171 of A172 3H cholesterol mix 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 sample 59 60 61 62 64 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 85 86 87 88 89 91 92 94 95 96 97 98 100 101 103 104 105 106 107 109 110 112 113 114 115 116 118 119 compound concentration PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M PFBS 1 M PFBS 1 M PFBS 1 M PFBS 10 M PFBS 10 M PFBS 100 M PFBS 100 M PFHS 1 M PFHS 1 M PFHS 1 M PFHS 10 M PFHS 10 M PFHS 100 M PFHS 100 M PFOS 1 M PFOS 1 M PFOS 1 M PFOS 10 M PFOS 10 M PFOS 100 M PFOS 100 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.003 M Rosuvastatin 0.03 M Rosuvastatin 0.03 M Rosuvastatin 0.3 M Rosuvastatin 0.3 M 14C Label 3H-cholesterol 14C-cholesterol 14C-cholesterol corrected g protein 14C-cholesterol % of control Average SD (dpm) (dpm) for recovery (dpm) per mg protein (dpm/mg) % of control 14C butyric acid 11671 3180 5470 348.0 15718 126.4 14C butyric acid 11805 4589 7804 337.9 23097 185.7 14C butyric acid 11641 3479 6000 345.7 17357 139.6 175.7 64.8 14C butyric acid 11648 4837 8337 316.4 26346 211.9 14C butyric acid 14C butyric acid 11906 12251 2793 3204 4710 5250 344.1 379.8 13687 13825 110.1 111.2 110.6 0.8 14C butyric acid 12599 1926 3069 403.5 7607 61.2 71.1 15.9 14C butyric acid 10509 2215 4231 380.6 11117 89.4 14C butyric acid 12593 2141 3413 437.8 7797 62.7 14C butyric acid 12245 2079 3409 412.7 8259 66.4 73.4 8.4 14C butyric acid 11423 2076 3649 412.6 8843 71.1 14C butyric acid 12013 2892 4833 469.7 10290 82.7 14C butyric acid 11885 2655 4485 356.5 12579 101.2 92.1 10.1 14C butyric acid 14C butyric acid 11791 11061 2641 2162 4497 385.2 11673 93.9 3924 388.3 10107 81.3 14C butyric acid 12385 137 222 387.5 573 4.6 4.8 0.7 14C butyric acid 12579 127 203 395.7 512 4.1 14C butyric acid 12107 162 269 387.5 693 5.6 14C butyric acid 12774 35 55 399.5 138 1.1 1.3 0.2 14C butyric acid 11196 33 59 392.7 151 1.2 14C butyric acid 12355 43 70 366.0 191 1.5 14C butyric acid 13007 30 46 382.3 121 1.0 1.2 0.4 14C butyric acid 12240 43 71 371.9 190 1.5 14C palmitic acid 14199 978 1383 332.2 4163 60.6 56.3 7.1 14C palmitic acid 12961 745 1154 349.3 3304 48.1 14C palmitic acid 13307 963 1453 350.8 4142 60.2 14C palmitic acid 13107 863 1322 360.9 3663 53.3 40.8 17.6 14C palmitic acid 12361 429 697 357.3 1950 28.4 14C palmitic acid 14351 1098 1536 341.4 4500 65.5 62.4 4.3 14C palmitic acid 12742 954 1503 368.2 4082 59.4 14C palmitic acid 13653 1536 2259 358.7 6296 91.6 84.3 7.1 14C palmitic acid 12703 1392 2200 381.5 5766 83.9 14C palmitic acid 12038 1184 1975 371.2 5320 77.4 14C palmitic acid 13137 1031 1576 398.7 3952 57.5 58.2 1.1 14C palmitic acid 11960 991 1663 410.1 4056 59.0 14C palmitic acid 11398 1551 2732 407.0 6712 97.6 82.3 21.7 14C palmitic acid 14337 1171 1640 356.6 4599 66.9 14C palmitic acid 11689 851 1462 426.9 3424 49.8 57.4 8.1 14C palmitic acid 13496 1062 1580 407.0 3881 56.5 14C palmitic acid 13069 1193 1833 404.8 4527 65.9 14C palmitic acid 13862 1377 1994 393.5 5068 73.7 71.1 3.7 14C palmitic acid 14202 1377 1947 413.1 4712 68.5 14C palmitic acid 9771 742 1525 420.8 3623 52.7 70.7 25.5 14C palmitic acid 8624 980 2281 374.2 6097 88.7 14C palmitic acid 6247 152 488 422.3 1157 16.8 11.7 4.4 14C palmitic acid 8729 122 281 470.2 597 8.7 14C palmitic acid 10667 128 241 363.1 663 9.7 14C palmitic acid 6797 127 375 408.7 918 13.4 14.3 1.3 14C palmitic acid 4697 107 457 437.5 1045 15.2 14C palmitic acid 9054 141 313 449.1 696 10.1 15.1 7.0 14C palmitic acid 4674 125 537 390.9 1374 20.0 mix 1 mix 2 input 3H cholesterol 21387 21723 input 3H cholesterol input 3H cholesterol 20596 19529 17423 21082 average input 3H cholesterol 20504 20076 04-12-2006 Final version Appendix to TNO-report of 3M-02 Appendix to TNO-report of 3M-02 Final version 04-12-2006 Average [14C]-Cholesterol synthesis in rat hepatocytes (% of control) Concentration: experiment 1 2 3 4 AVERAGE SD experiment 1 2 3 4 AVERAGE SD experiment 1 1 M 10 M 100 M 0.003 M 14C acetate incorporation, n=4, PFBS 85.0 82 76 115 157 177 33 34.9 42.1 90 67.3 93.7 80.8 85.3 97.2 34.4 51.7 57.3 14C acetate incorporation, n=4, PFHS 91 82 84 148 136 111 63.8 38.8 35.9 91.8 154.3 100 98.7 102.8 82.7 35.4 52.5 33.1 14C acetate incorporation, n=4, PFOS 76 84 65 2 3 4 AVERAGE SD experiment 1 2 3 4 AVERAGE SD 130 119 132 43.3 32.6 38.4 139.9 113.9 89 97.3 87.4 81.1 45.7 39.6 39.7 14C acetate incorporation, n=4, Rosuvastatin 45 40.0 6.2 3.8 23.8 21.8 0.03 M 11 8 1 1 5.4 4.9 0.3 M 3 2 0.5 0.6 1.5 1.2 Concentration: experiment 1 2 3 4 AVERAGE SD experiment 1 2 3 4 AVERAGE SD experiment 1 2 3 4 AVERAGE SD experiment 1 2 3 4 AVERAGE SD 1 M 10 M 100 M 0.003 M 14C butyrate incorporation, n=4, PFBS 61.0 128 92 24 146 140 175 37.9 50.7 60.3 92 52.2 118.7 84.2 92.7 111.5 46.8 47.9 48.6 14C butyrate incorporation, n=4, PFHS 115 101 100 190 220 200 45.4 48.3 51.9 124.5 175.7 110.6 118.7 136.3 115.6 59.2 76.5 61.8 14C butyrate incorporation, n=4, PFOS 76 98 58 262 223 286 29.1 68.8 45.5 71.1 73.4 92.1 109.6 115.8 120.4 103.8 72.6 112.1 14C butyrate incorporation, n=4, rosuvastatin 24 46.0 3.7 4.8 19.6 19.9 0.03 M 5 5 16 1 1 5.8 7.0 0.3 M 3.0 3 4 0.5 1.2 2.2 1.6 Concentration: experiment 1 2 3 AVERAGE SD 1 2 3 AVERAGE SD 1 2 3 AVERAGE SD 1 2 3 AVERAGE SD 1 M 10 M 100 M 0.003 M 14C laurate incorporation, n=3, PFBS 84.0 97 87 131 102 100 68.5 77.2 53.3 94.5 92.1 80.1 32.5 13.1 24.1 14C laurate incorporation, n=3, PFHS 108 102 68 150 103 134 74.4 65.4 64.2 110.8 90.1 88.7 37.9 21.4 39.2 14C laurate incorporation, n=3, PFOS 113 85 65 119 124 125 100.4 77.6 55.6 110.8 95.5 81.9 9.5 24.9 37.6 14C laurate incorporation, n=3, rosuvastatin 25 35.0 7.5 22.5 13.9 0.03 M 7 15 1 7.8 6.8 0.3 M 3 3 0.5 2.2 1.4 Concentration: experiment 1 3 4 AVERAGE SD 1 3 4 AVERAGE SD 1 3 4 AVERAGE SD 1 3 4 AVERAGE SD 1 M 10 M 100 M 0.003 M 14C palmitate incorporation, n=3, PFBS 76.0 56 71 50.4 54.6 37.1 56.3 40.8 62.4 60.9 50.5 56.8 13.4 8.4 17.6 14C palmitate incorporation, n=3, PFHS 63 68 59 40.2 45.7 47.5 84.3 58.2 82.3 62.5 57.3 62.9 22.1 11.2 17.7 14C palmitate incorporation, n=3, PFOS 90 62 51 41.6 63.1 34.5 57.4 71.1 70.7 63.0 65.4 52.1 24.7 5.0 18.1 14C palmitate incorporation, n=3, rosuvastatin 48 11.7 11.7 23.8 21.0 0.03 M 20 7 14 13.8 6.5 0.3 M 12 5 15.1 10.7 5.2 Page A172 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