Document 3QnaeO0n9axyoOV9xar7JmxM6

-- ' Commercial-in-Confidence Chemex reference : ENV8010/020737 P. 1 37 ,g ~ The growth inhibition of the marine alga Skeletonstatum by C ~ Report for Notox B.V - - _ -; ' Report issued by : Sponsor: Chemex Environmental International Limited Notox B .V Unit J Hambakenwetering 7 Broad Lane Industrial Estate P .O . Box 3476 Cottenham 5203 D L Cambridge `s-Hertogenbosch CB24 8SW The Netherlands England April 2007 ENV8010/020737 Page 1 of 18 p. 2 ~ p. 3 Contents Page Compliance with Good Laboratory Practice standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3 Quality Assurance audit statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5 1 . Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2 . Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6 2 .1 Test mate rial . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2 .2 Test organism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7 2 .3 Preparation of test solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7 2 .4 Test methods and conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 3 . Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9 3 .1 Water quality data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9 3 .2 Percent inhibition by biomass integral and growth rate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9 3 .3 EC5o values by biomass integral (EbC50 value) and growth rate (ErC50 value) . . . . . . . . . . .9 4 . Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9 5 . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10 Graph s 1 . Growth curves for control and test concentrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11 2 . Estimation of 0 to 48 hours EbC50 value . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12 3 . Estimation of 0 to 72 hours EbC50 value . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . : . . . . . . . . . . . .13 4 . Estimation of 0 to 48 hours EiC50 value . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14 5 . Estimation of 0 to 72 hours ErC50 value . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15 Appendice s 1 . Cell density measurements - test sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16 2 . Reference material results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17 3 . Culture media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18 ENV8010/020737 Page 2 of 1 8 ~ p. 4 Compliance with Good Laboratory Practice standard s I, the undersigned, hereby declare that the study described in this report was performed under my, supervision, and that the final report fully and accurately reflects the raw data generated during the conduct of the study, in compliance with international codes of Good Laboratory Practice including : Section II of Annex 1 to the European Parliament and council Directive 2004/10/EC and Annex 1 to the European Parliament and council Directive 2004/9/EC (Official Journal No . L 50) and embodied within : The UK Good Laboratory Practice Regulations 1999 (The United Kingdom GLP Regulations 1999, Statutory Instrument 3106) as amended by: The UK Good Laboratory Practice (Codification Amendments Etc .) Regulations 2004 (Statutory Instrument No 994) These principles are in accordance with the OECD Principles of Good Laboratory Practice, revised 1997 (ENV/MC/CHEM(98)17) . ~'. ~. ... .... .. ~~ Tom Fisher Date Study Director A1) ~~ .. .... ... ENV8010/020737 Page 3 of 18 p. 5 Quality Assurance audit statemen t Studies of this type are subject to random process-based inspections by the Chemex Quality Assurance Unit . Date of relevant inspection : 13 & 16 February 2007 Date relevant inspection reported : 16 February 2007 Date of study plan inspection : 28 February 200 7 This report has been audited by the Chemex Quality Assurance Unit . It is considered to be an accurate description of the procedures and practices employed during the course of the study and an accurate presentation of the findings . Date audit started : 20 April 2007 Date audit reported : 23 April 2007 Signed : .. ..... ... ... . .. . .. . ... .. ... . .. .. ... ..... David S tittl e Quality Manager Date : . . . . . ~:.~. . .~,~r`i ~ . . .2:Q-7 . . . . . . . . . . . . . . . . . . . . . . . . . . All reports are individually audited . Reports are issued to Management on a monthly basis . Raw data used to derive results included in this report, supplied by either the sponsor or subcontractor (where applicable), are not audited by Chemex Environmental International Limited . ENV8010/020737 Page 4 of 1 8 S p. 6 S~tA #*fi7 Z CC.( Summary This section summarises aquatic toxicity test results obtained by Chemex Environmental International Limited on a sample as detailed below : Test commissioned by : Notox B . V Substance under test: : Samp Chemxrfnc le : EC002073 7 Study : ENV801 0 Test species : Skeletonema costatum, strain 1077/5 Test type : Acute toxicity : 72 hour EC5 0 Protocol : Static test conditions according to SOP E209 based on ISO/DIS BS EN ISO 10253 : 1998 "Water quality - Marine Algal Growth Inhibition Test with Skeletonema costatum and Phaeodactylum tricornutum. Test period : From 2 to 5 April 200 7 Test concentrations : 0, 2 .2, 4 .5, 10, 22, 45 and 100mg/1 Test performed at : Chemex Environmental International Limited Unit J Broad Lane Industrial Estate Cottenham Cambridge CB24 8SW England Results : Period of EbCSa value mg/l Period of E , C50 value mg/I exposure biomass exposure growth rate hours (95% confidence limits (hours) 95% confidence limits Oto48 0 to72 14mg/1 23m 1 10-19m 1 Oto48 ~ ) (21 - 25m 1) 13mg/1 10-17m 1 0 to72 24m ~1 ) (21 - 27m 1 NOECb (biomass) NOECr 4 .5mg/1(0-72 hours) (growth rate) ENV8010/020737 Page 5 of 18 p. 7 s~ ~ f7'Z 1 . Introductio n This report contains a description of the methods used and the results obtained during a study to investigate the marine algal growth inhibition ofC -J to Skeletonema costatum . The objective of this study was to determine the 72 hour median effective concentration (EC50) which is defined as the concentration which inhibits the growth of the marine algae by 50% after a 72 hour exposure period at 20C . 2. Materials and method s Unless otherwise specified all methods mentioned in this report are according to Chemex Environmental International Limited standard procedures . All records of measurements and observations made during this test will be collated and held in the Chemex Environmental International Limited archives at Unit J, Broad Lane Industrial Estate, Cottenham, Cambridge CB24 8SW, England . 2 .1 Test material Identification : Supplied by: Notox B . V Date of receipt : 27 February 2007 Chemex reference : EC002073 7 Required storage conditions : Store in original containers at room temperature Actual storage conditions : Room temperature in dark Quoted stability : Stable Quoted appearance : Amber viscous liquid/paste Observed appearance : Amber viscous liquid/paste Quoted odour : Not Stated Quoted solubility in water : Complete ENV8010/020737 Page 6 of 18 / p. 8 s~Uj i f) Z CGC. 2.2 Test organis m Species : Skeletonema costatum strain : 1077/5 Source : Culture Collection of Algae and Protozoa Dunstaffnage Marine Laboratory, Scotlan d Culture conditions : Temperature : 20 1 C Illumination : 6000 - 10000 lux continuous white light Shaking : 200rpm Culture media: Natural seawater with added nutrients according to the ISO standard, with concentrations of iron and EDTA at the levels recommended by VKI (Danish Water Quality Institute) (see Appendix 2) . 2 .3 Preparati on of test solutions The behaviour oi C ':] was examined in seawater according to the method described in the Harmonised Offshore Chemical Notification Format (OSPAR 1995) . A nominal 1,000mg/1 solution was prepared in dilution water (see section 2 .4), shaken vigorously and allowed to stand for one hour . A faintly grey solution was obtained . Test concentrations were prepared by mixing specified quantities of the sample with culture medium to obtain the required concentrations . A preliminary study had identified the 72 hour EC50 as being between 10 and 32mg/1 and therefore definitive test concentrations were prepared as 0, 2 .2, 4 .5, 10, 22, 45 and 100mg/l . ENV8010/020737 Page 7 of 1 8 9 P. 9 2 .4 Test methods and condition s Static test conditions according to ISO draft method (1991) "WATER QUALITY Marine algal growth inhibition test with Skeletonema costatum and Phaeodactylum tricornutum" (ISO/DIS 10253) . Chemex SOP reference : E209 "Marine Algal Growth Inhibition test with Skeletonema costatum an d Phaeodactylum tricornutum" Test period : 2 to 5 April 200 7 Test volume : 200ml Test vessel : 250m1 conical flas k Number of replicates : 6 control flasks, 3 replicates at each test concentration Seawater : Collected on 6 December 2006 from the CEFAS laboratories at Burnham-on-Crouch directly from the River Crouch estuary . Algal test inoculunn : From a pre-culture growing in exponential phase under conditions described in 2 .2 above . Inoculum level adjusted to give an initial cell density of 1 x 104 cells/ml . Test conditions : Temperature : 20 1 C Illumination : 6000 - 10000 lux continuous white light Shaking : 200 rpm Water quality measurements : The pH (to 0 .1) and temperature (to 0 .5C) were measured on the pooled replicates for each test and control solution immediately prior to initiating the test and at the end of the full 72 hour test period . Observations/frequency : The cell density was determined at 24, 48 and 72 hours on a small volume removed from each replicate flask . The cell counts were made using a haemocytometer and microscope . Calculation and expressio n of results : Growth curves for each test concentration were plotted as logarithm of the mean cell density against time . The inhibition of growth was calculated using both the biomass (area under the growth curve) and the growth rate method . Where possible, the EC50 values were estimated graphically and 95% confidence limits calculated according to the method of ToxCalcTM Version 5 .0 "Comprehensive Toxicity Data Analysis and Database Software", copyright 1994-1996 . ENV8010/020737 Page 8 of 1 8 ~ P. 10 3. Results 3 .1 Water quality data Nominal concentration Mean H values Temperature OC (mg/1) Sta rt of test End of test Start of test End of test 0 8 .0 8 .6 20 .0 20 .0 2 .2 8 .0 8 .7 20 .0 20 .0 4 .5 8 .0 8 .7 20 .0 20 .0 10 8 .0 8 .3 20 .0 20 .0 22 8 .0 8 .0 20 .0 20 .0 45 8 .0 8 .0 20 .0 20 . 0 1 100 8 .0 8 .0 20 .0 20 .0 3 .2 Percent inhibition by biomass integral and growth rate Nominal concentration Percent inhibition Percent inhibition biomass integral by growth rat e (mg/1) by 48 hours 72 hours 48 hours 72 hours 2 .2 -6 -8 2 -3 4 .5 0 2 -1 1 10 39 45 13 17 22 68 73 32 35 45 100 100 100 100 100 100 100 97 10 0 Note : Negative inhibition values indicate an increase in growth compared with controls . 3 .3 EC5o values by biomass integral (EbCSO value) and growth rate (ErC5a value) Period of EbCso value mg/1 Period of E, C50 value mg/1 exposure biomass exposure growth rate hours (95% confidence limits) hours (95% confidence limits 14mg/1 23mg/1 0 to 48 (10 - 19m 1 0 to 48 (21 - 25m 1 ) 13mg/1 24mg/1 0 to 72 10 - 17m 1 0 to 72 (21 - 27m 1 NOEC b (biomass) NOECr (growth rate ) 4 .5mg/1(0-72 hours) 4. Discussio n ENV8010/020737 Page 9 of 1 8 DI P. 11 S~tvt i ~iZ ~G( The 72 hour EC50 o_C ~ io Skeletonema costatum was estimated as 13mg/1 using the biomass integral and 24mg/1 by growth rate calculation . The 48 hour EC50 was 14mg/1 by biomass integral and 23mg/1 by growth rate . The 72 hour NOEC was estimated as 4 .5mg/1 as determined by the Bonferroni T test . The growth curves illustrated in Graph 1 show that growth inhibition increases with increasing concentration, demonstrating a clear dose response to the test material . The inhibition values given in 3 .2 support these observations . The test organism sensitivity was also monitored during this study using the recommended reference substance 3,5 DCP . The algae were exposed to a 1 .5mg/1 concentration of 3,5 DCP for 72 hours under test incubation conditions and therefore the 42% inhibition by growth rate represents an acceptable level of sensitivity (ie . 72 hour inhibition was within the range of 20 to 80%) . The water quality measurements of the test solutions are given in 3 .1 and were within acceptable limits. The cell density of the control during the 72 hour test period increased by a factor of 67, which is satisfactory . There was no chemical analytical confirmation of the actual dissolved concentrations . The dissolved concentrations were likely to be as given in this report as the 'sample was soluble in water . 5. Reference s ISO/DIS BS EN ISO 10253 : 1998 "Water quality - Marine Algal Growth Inhibition Test with Skeletonema costatum and Phaeodactylum tricornutum . VKI - "Addendum to PARCOM Ring Toxicity Test with Marine Unicellular Algae" (1991) (Danish Water Quality Institute) . OSPAR - Oslo and Paris Conventions for the Prevention of Marine Pollution Programmes and Measures Committee (PRAM) Harmonised Offshore Chemical Notification Format 1995 . ToxCalcTM Version 5 .0 "Comprehensive Toxicity Data Analysis and Database Software",copyright 1994-1996 . ENV8010/020737 Page 10 of 18 ~/ p. 12 Graph 1 Growth curves for control and test concentrations 1 .0 0 0 .75 0 0 0 .-; 0 .50 E V 0 .25 0 .00 0 24 48 Exposure period (hours) 72 Concentration in mg/1 --6--0 -2 .2 - 4 .5 - 10 -22 -45 -10 0 Statistical data NOECb (biomass) = 4 .5mg/1 NOECr (growth rate) = 4 .5mg/1 LOEC = 10mg/1 As determined by the Bonferroni T test, based on the 0-72 hour values . ENV8010/020737 Page 11 of 1 8 /C~ p. 13 Graph 2 Estimation of 0 to 48 hours EbC5o value 1.0 0 .9 ; 0.8 0.7 0.6 Z 0.5 0.4 0.3 0.2 0 .1 0 .0 1 10 100 1000 Concentration mg/1L ---------- Denotes 95% confidence limit s Statistical data Estimated EC50 value = l4mg/ l (as determined by the Maximum Likelihood-Probit Method) 95% confidence limits = 10 to 19mg/ 1 ENV8010/020737 Page 12 of 1 8 13 p. 14 Graph 3 Estimation of 0 to 72 hours EbCso value 1 .0 0 .9 0.8 . 0.7 c 0. 6 ~' ; ; c 0.5 ^ ; 0 0.4 O CL 0 .3 0 .2 0. 1 0 .0 1 10 100 1000 Concentration mg/L ---------- Denotes 95% confidence limits Statistical data Estimated EC50 value = l3mg/ l (as determined by the Maximum Likelihood-Probit Method) 95% confidence limits = 10 to 17mg/ 1 ENV8010/020737 Page 13 of 18 /~ p. 15 Graph 4 Estimation of 0 to 48 hours ErC5o value 1 .0 0 .9 0 .8 0. 7 0.6 0.5 0 .4 0 .3 0 .2 0 .1 0 .0 1 10 100 Concentration mg/L Statistical data Estimated EC50 value = 23mg/1 (as determined by the Spearman-Karber Method) 95% confidence limits = 21 to 25mg/1 ENV8010/020737 Page 14 of 18 /J p. 16 Graph 5 Estimation of 0 to 72 hour E,.C50 value 1.0 0.9 0.8 0 .7 0 .6 0 .5 ea 0 .4 0 .3 0 .2 0.1 0.0 1 10 100 Concentration mg/L Statistical data Estimated EC50 value = 24mg/ 1 (as determined by the Spearm an-Karber Method) 95% confidence limits = 21 to 27mg/ l ENV8010/020737 Page 15 of 18 p. 17 Appendix 1 Cell density measurements - test sample Concentration Cell density measurements ~m~~ Replicate cells/ml x 104 24 hours 48 hours 72 hours 1 2 .67 22 .67 69 .33 2 6 .33 21 .67 62 .33 3 6 .67 27 .67 72 .0 0 0 4 4 .00 24 .00 56 .67 5 4 .33 23 .00 67 .67 6 8 .33 22 .67 73 .3 3 Mean 5 .39 23 .61 66 .89 1 10 .00 22 .00 87 .33 2 .2 2 5 .33 24 .67 71 .0 0 3 6 .00 19 .67 68 .33 Mean 7.11 22 .11 75 .55 4 .5 1 2 3 4 .67 3 .67 6 .67 29 .33 23 .33 20 .33 66 .6 7 60 .00 64 .33 Mean 5 .00 24 .33 63.6 7 1 4 .00 18 .00 35 .00 10 2 3 2 .00 13 .00 32 .67 4 .00 15 .67 30 .6 7 Mean 3 .33 15.56 32 .78 22 1 0 .00 5 .33 2 3 2 .33 14 .67 3 .00 8 .00 Mean 1 .78 9 .33 14 .33 14 .6 7 16 .33 15.11 45 1 2 3 0 .00 1 .33 0 .67 0 .00 0 .00 0 .00 0 .0 0 0 .00 0 .00 Mean 0 .67 0 .00 0 .0 0 1 0 .00 0 .00 0 .00 100 2 0 .00 0 .00 0 .00 3 0 .00 1 .33 0 .0 0 Mean 0.00 0.44 0 .00 Me an initial cell density : 1 x 104 cells/ml . ENV8010/020737 Page 16 of 1 8 17 p. 18 Appendix 2 Reference material result s The test organism sensitivity was monitored du ring this study using the recommended reference subst an ce 3,5 DCP . The algae were exposed to a 1 .5mg/1 concentration of 3,5 Dichlorophenol for 72 hours under test incubation conditions. Percent inhibition by biomass integral and growth rate inhibitio n Nominal Percent inhibition Percent inhibition concentration by biomass integral by growth rate 3,5-DCP 48 hours 72 hours 48 hours 72 hours 1 .5m /1 60 75 38 42 Cell density measurements Cell density measurements 3C ,5-oDCnPc(megn/1t)ration Replicate cells/ml x 104 24 hours 48 hours 72 hours 1 2 .67 22 .67 69 .33 2 6 .33 21 .67 62 .33 0 3 6 .67 27 .67 72 .0 0 4 4 .00 24 .00 56 .67 5 4 .33 23 .00 67 .67 6 8 .33 22 .67 73 .3 3 Mean 5 .39 23 .61 66.89 1 6 .00 9 .67 17 .33 1 .5 2 3 .67 3 2 .33 Mean 4 .00 7 .67 13 .00 5 .00 7 .00 7 .45 12 .44 ENV8010/020737 Page 17 of 1 8 / Y P. 19 Appendix 3 Culture medi a Culture medium : Prepared in natural seawater according to the ISO/DIS 10253 standard except that concentrations of iron and EDTA were at the levels recommended by VKI (Danish Water Quality Institute) . Seawater : Filtered, and sterilised by autoclaving at 120C for 30 minutes . Method of preparation : Sterile nutrient stock solutions were prepared and added to the seawater to obtain the culture medium . Composition of culture medium : Nutrient Final concentration in test solution FeC13 6H20 16 .4g/1 (Fe) MnC12 .4H20 605g/l (Mn) ZnSO4 .7 H20 150g/1(Zn) CuSO4 .5 H20 0 .6g/l (Cu) CoC12 .6 H20 1 .5g/1 (Co) 1-131303 17 .1mg/1 NaZ EDTA, 2H2 0 100g/1 Thiamin hydrochloride 25 g/1 Biotin 0 .005g/1 B12 (cyanocobalamin) 0 .05g/1 K3 P041-120 3 .25mg/1 NaNO3 50 .0mg/1 NazSiO3 .5Hz0 14 .9mg/1 ENV8010/020737 Page 18 of 1 8 ~9 This TSCA 8(e) substantiation applies to the following studies for materials that have never been commercialized in the U .S.: [test substance ] In addition to the chemical identities or formula of a proprietary mixture, 3M maintains its 3M proprietary ID #s and names of 3M personnel . 1 . Yes, the CBI claim is on behalf of 3M Company, including its wholly-owned subsidiaries and affiliates . 2. The period of time for 3M's confidentiality claim is indefinite . 3M's claim is for the chemical identities of components of materials that have been commercialized in the U.S . 3M engaged in significant R&D efforts to develop this product . 3M has always claimed the components of the chemical identities in question as proprietary and continues to do so today . With this knowledge, competitors could do one or more of the following : (a) deduce (and potentially duplicate) 3M's process technology, process chemistry and process know-how; (b) develop products with components and/or functionalities identical or similar to 3M's current products ; and (c) devise marketing strategies and tactics to promote their current products as compared to 3M's current products . In these ways, disclosure of 3M's proprietary information would deprive 3M of its intellectual property and its competitive advantage and would bestow a competitive advantage that 3M's competitors would not otherwise be able to achieve using their independent "know how" . 3M would be deprived of research and development investment return as a result of such disclosure . 3 . If a PMN was written, the PMN number is provided in the confidential copy of the Index to Studies Submitted . Disclosure would have been made to the U .S . EPA . Otherwise, no other disclosure was made. 4 . The exact chemical identities or formula of the proprietary mixture has been kept confidential using general descriptions (e .g., experimental product) on all hazard communication (MSDS and labels) . In addition, the measures taken to prevent undesired disclosure include strict R&D 3M security procedures with disclosure of the chemical identity of the substance within 3M on a need to know basis . 5 . Contract employees, engineering and construction firms and other third parties with access to this information sign a confidentiality agreement or are otherwise bound through agency that prohibits disclosure of 3M business confidential information . 6 . No, the chemical identities only appear as a generic name on the MSDS . 7 . No . 8 . Refer to response to Question #2 . 9 . No . 10 . a) Yes b) Approximately 3 years . C) The mixture is a raw material used at a very small percentage in hard surface coating formulations . Competitors are not aware of the exact formulation of the mixture . 11 . Determining the exact chemical identity of the substance would require the use of very sophisticated analytical equipment and methods by highly skilled chemists and would require an extended period of time . 12 . a.) Yes, refer to response to Question #2 . b.) Yes, refer to response to Question #2 . c.) Yes, refer to response to Question #2 . 13 . No CAS number exists for the polymer in the product . 14 . No . p . 21 3M Compan y Contact name : Deanna Luebker, PhD, 3M TSCA 8(e) Coordinato r 3M Toxicology Assessment and Compliance Assurance 3M Center 220-06-E-03 St. Paul, MN 5514 4 651-737-1374 (office) 651-733-1773 (fax) d1luebkergmmm .com