Document pBEow2aqexBbJV66nZRNqnZ1j

PERFLUOROOCTANESULFONYL FLUORIDE (POSF; T-7661.4) TOXICITY STUDY BY INHALATION ADMINISTRATION TO CD RATS FOR 13 WEEKS FOLLOWED BY A 4 WEEK RECOVERY PERIOD Volume 2 MIN 313/023622 PERFLUOROOCTANESULFONYL FLUORIDE (POSF; T-7661.4) TOXICITY STUDY BY INHALATION ADMINISTRATION TO CD RATS FOR 13 WEEKS FOLLOWED BY A 4 WEEK RECOVERY PERIOD Volume 2 Sponsor 33BMUSMMtuSNPiAlaCCd5.uieo5nlnr,1gpt3eo23rr2,-a30t2e-22TE0o,-x0i2c,ology, Research Laboratory HWAHCPEENaluuocm2nnGoo8ttbnliiLlnnrb4eAigguHydddNrgSRyooeD,,nnos,.haLdiri,efe, Sciences Ltd., Report issued: 28 September 2005 Page 366 of 740 MIN 313/023622 CONTENTS Page CONTENTS................................................................................................................................... 2 COMPLIANCE WITH GOOD LABORATORY PRACTICE.................................................... 4 QUALITY ASSURANCE STATEMENT................................................................................... 5 CONTRIBUTING SCIENTISTS.................................................................................................. 6 SUMMARY................................................................................................................................... 7 INTRODUCTION......................................................................................................................... 9 EXPERIMENTAL PROCEDURE................................................................................................ ANTPDSQSATEENEAUURSTCITAMTIDHRALAYSAOOILTTULPLSRYOBSOCMEBYSAHGATASSAEYTAENSDN.MRNU.A.UDV.CRE.GL.A.EHAN.EE.TI.NATM.SA.I.C.NTO..EN..E.OD.NN..D...L.ATSF....OS.N..O.....T....GD...R.....R....YM....A..U............UR..C............C.LT..........A.H....U..........TI....R.....V.....I....E..O...I.......N......N...........G...........................P...............R....................O.....................C..................E..................D.....................U.....................R....................E.................S.................................................................................................................................................................................................................................................................................................................................................................................... 11 2221111122516392 RESULTS...................................................................................................................................... 27 DISCUSSION................................................................................................................................ 35 REFERENCES.............................................................................................................................. 36 FIGURES 1. Bodyweights - group mean values.......................................................................................... 37 : 367 : TABLES MIN 313/023622 Page 23456781........ BHBUFOMMooiarraiooigdecnccdamyrrahonowlcaesyswotmeccsonoiioeglissposiphugtig-irctmhcysygtppprs---oatag-itggutohrhgrrponooooruloumul-opopupggegpmaymrmyonme-eeu-avaaepggnnnaarrmnolvovvuuuaevaaepalpllasunuudl.de.ueeiv.si.ssses..at....tsr....lr.....iui.....bb.....e.....uu.....s.....t..t......i...i......oo............n.n................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................... 4437658801826037 APPENDICES 42357968111.........01.. DWBABHUFIpCnHoioabardeoeoiidesniellcvdvomylykahailwpllcduaelpyyuromtuetosooeenipasilglissslthiosoh-futfy-rpgdertmogssyyaroiraiantpsc-hn--ttdetaieoiioiiilwnronnvolnmodendidbedgixfiiiisv-vuvngiiaeciniaiahimrdddandlvltuluuisdaivunanaafit-alraviillgoinltiivvnvuisndndoa.eaadeus.inll..lnsiau..u-uv...lg-e...ieei...vssdni...ss.n....aud.........d.....lai.......u..v.i....l......v.e...i....v......id.s.......da.......u........u.l.......a.u.......a...l.......e.....l.........f.s........fi.........in............n.........d.............d.........i.........i.n..........n.........g.........g.........s.........s............................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................................ 222211111110991352713032186028858 ADMINISTRATION OF POSF BY INHALATION TO RATS.................................................... 298 MSCICARNONAINNGAELLYESCIST.R...O...N....M...I..C..R...O...S..C...O...P..E....E..X...A...M....I.N...A...T..I.O...N....A...N...D....X...-.R...A...Y.................................. 369 ANALYTICAL PHASE REPORT.................................................................................................. 518 HEYUENTRIENSGEDAORNCHRECSEENATRRCEHGCLPENCTORMEPGLLIAPNCCOEMSPTLAIATNECMEESNTTAST...E...M...E...N...T..S.................................................... 773385 Volume 1........................................................................................................................................... 1 Volume 2........................................................................................................................................... 366 : 368 : MIN 313/023622 SCANNING ELECTRON MICROSCOPE EXAMINATION AND X-RAY MICROANALYSIS Plymouth Electron Microscope Unit Scanning Electron Microscope Examination and X-Ray Microanalysis. Study Number: MIN313R Huntingdon Life Sciences HCaumntbinsgdon PE284HS Plymouth Electron Microscope Unit University of Plymouth PDlryamkeoCutihrcus P L 4 8AA 01752 233092 : 369 : MIN 313/023622 Materials and Methods 1. SEM examination of urinary bladder tissue samples: Subsamples (approximately 0.5 x 0.5cm) were cut from fixed tissue samples (supplied in 70% ewtihtahncooll)loainddalcsriitlivcearl apdohienstivderi,esdpu(pttreortococaotleadttwacithhedg)o.ldDaryndsaemxapmleisnewdeirne amJoEuOnLted56o0n0sSpEecMimaetn15stkuVb.s Four images, each at 500x magnification, were collected to illustrate typical epithelial morphology. 2. SEM/X-Ray microanalysis ofurine samples: Dry samples as supplied were carbon coated and examined in a JEOL 6100 SEM at 15kV. X-ray data were collected and analysed with an Oxford Instruments ISIS X-Ray Analysis system. : 370 : MIN 313/023622 Specimen Preparation for Scanning Electron Microscopy Critical Point Drying (CPD) 1. Fix freshly excised sample appropriately (eg. 2.5% Glutaraldehyde in 0.1M phosphate buffer) 2. Wash in buffer - 3x 15 minutes 3. Dehydrate in ethanol series 30% -15 minutes 50% - 15 minutes 70% -15 minutes 90% -15 minutes 100% - 15 minutes 100% -15 minutes 4. Fill CPD chamber with 100%ethanol and cool to 10C or less 5. Place specimens in CPD chamber 6. Close and flood with liquid carbon dioxide 7. Exhaust alcohol from chamber. 8. Leave sample 30 minutes in chamber to infiltrate with carbon dioxide. 9. Exhaust remaining ethanol 10. Heat chamber to critical point 11. Vent carbon dioxide gas from chamber 12. Remove sample from chamber and mount for SEM examination. : 371 : MIN 313/023622 Results 1. SEM Examination ofUrinary Bladder Epithelium. Sample 19 - only two images were taken. The epithelium had separated from the sample, no cells remained and image quality, under the operating conditions used for the study, were inappropriate to produce adequate images ofthe sample. : 372 : MIN 313/023622 : 373 : MIN 313/023622 : 374 : MIN 313/023622 : 375 : MIN 313/023622 : 376 : MIN 313/023622 : 377 : MIN 313/023622 : 378 : MIN 313/023622 : 379 : MIN 313/023622 : 380 : MIN 313/023622 381 MIN 313/023622 : 382 : MIN 313/023622 : 383 : MIN 313/023622 : 384 : MIN 313/023622 : 385 : MIN 313/023622 : 386 : MIN 313/023622 : 387 : MIN 313/023622 : 388 : MIN 313/023622 : 389 : MIN 313/023622 : 390 : MIN 313/023622 : 391 : MIN 313/023622 : 392 : MIN 313/023622 : 393 : MIN 313/023622 : 394 : MIN 313/023622 : 395 : MIN 313/023622 : 396 : MIN 313/023622 : 397 : MIN 313/023622 : 398 : MIN 313/023622 : 399 : MIN 313/023622 : 400 : MIN 313/023622 401 MIN 313/023622 2. SEM/X-Ray analysis of Urine samples Representative images are provided from the supplied dried samples to illustrate typical crystal structure and deposit morphology. X-Ray analysis spectra from the supplied dried samples (labelled MIN 313R), to show general elemental composition, and composition of specific crystals and deposits (as labelled) are provided. : 402 MIN 313/023622 : 403 : MIN 313/023622 : 404 : MIN 313/023622 : 405 MIN 313/023622 : 406 : MIN 313/023622 : 407 : MIN 313/023622 : 408 : MIN 313/023622 O perator: Plymouth EM Unit C lient: none J o b : MIN 313 Recoveiy cps control r (21/05/0212:19) 30-- 20--| 10H -r------1------ i------ 1 -- F T f 10 15 20 Energy (keV) : 409 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 1r (21/05/02 12:20) MIN 313/023622 : 410 : Operator ; Plymouth EM Unit Client : none Job : MIN 313 Recovery 4r (21/05/02 13:19) MIN 313/023622 : 411 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 5r (21/05/02 09:23) MIN 313/023622 : 412 : O perator: Plymouth EM Unit C lient: none Jo b : MIN 313 Recovery 6r (21/05/02 09:27) MIN 313/023622 : 413 : O perator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 7r (16/05/02 08:43) MIN 313/023622 : 414 : O perator; Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 8r (16/05/02 08:51) MIN 313/023622 : 415 : Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 9r (16/05/02 08:57) MIN 313/023622 : 416 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 10r (16/05/02 09:02) MIN 313/023622 Energy (keV) : 417 : Operator : Plymouth EM Unit Ciient : none Job : MIN 313 Recovery 11r (21/05/02 13:23) MIN 313/023622 : 418 : Operator : Plymouth EM Unit Client : none Job : MIN 313 Recovery 13r (21/05/02 09:31) MIN 313/023622 : 419 : O perator: Plymouth EM Unit CBent: none Job : MIN 313 Recovery 15r (21/05/02 09:35) MIN 313/023622 : 420 : O perator: Plymouth EM Unit CRent: none J o b : MIN 313 Recovery 16r (21/05/02 09:39) MIN 313/023622 : 421 : Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 20r (16/05/02 09:08) MIN 313/023622 : 422 : MIN 313/023622 Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 21 r (21/05/02 09:44) 20- 15- Cl 10- 1V n i i i | i i i------ r~ iii 10 "p*r r~ m` *t 1| 1 15 Energy (keV20) : 423 : O perator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 22r (21/05/02 09:50) MIN 313/023622 : 424 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 24r (21/05/02 09:55) MIN 313/023622 : 425 : O perator: Plymouth EM Unit C fient: none Job : MIN 313 Recovery 26r (16/05/02 09:17) MIN 313/023622 Is--I Energy (k : 426 : Operator: Plymouth EM Unit C lient: none Jo b : MIN 313 Recovery 27r (16/05/02 09:23) MIN 313/023622 : 427 : Operator : Plymouth EM Unit Client : none Job : MIN 313 Recovery 28r (16/05/02 10:35) MIN 313/023622 : 428 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 30r (16/05/02 10:40) MIN 313/023622 : 429 : O perator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 33r (21/05/02 10:00) MIN 313/023622 : 430 : O perator: Pfymouth EM Unit C lient: none J o b : MIN 313 Recovery 34r (21/05/02 10:04) MIN 313/023622 3 8-1 Cl V 10 15 Energy (k : 431 : Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 35r (21/05/02 10:09) MIN 313/023622 : 432 : O perator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 36r (21/05/0210:13) MIN 313/023622 : 433 : Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 38r (16/05/02 10:44) MIN 313/023622 : 434 : O perator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 39r (16/05/02 10:47) MIN 313/023622 : 435 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 40r (16/05/02 10:55) MIN 313/023622 : 436 : Operator: Plymouth EM Unit C lient: none Jo b : MIN 313 Recovery 42 background (15/05/02 09:53) MIN 313/023622 : 437 : Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 42 large crystal (15/05/02 09:56) MIN 313/023622 : 438 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 43R crystal (15/05/02 10:10) MIN 313/023622 : 439 : O perator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery cps 44r (15/05/02 10:17) MIN 313/023622 : 440 : Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 45r (15/05/0210:21) MIN 313/023622 : 441 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 46r (15/05/02 10:46) MIN 313/023622 : 442 : O perator; Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 50r (15/05/02 10:56) MIN 313/023622 : 443 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 51 r (15/05/02 11:02) MIN 313/023622 : 444 : O perator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 52r (15/05/0211:08) MIN 313/023622 : 445 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery ^ 53r (15/05/02 11:11) MIN 313/023622 : 446 : O perator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery ^ 55r (15/06/02 11:17) MIN 313/023622 : 447 : Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 58r (15/05/02 12:20) MIN 313/023622 : 448 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 59r (15/05/02 12:25) MIN 313/023622 : 449 : Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 60r (21/05/02 13:26) MIN 313/023622 Energy (kc : 450 : O perator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 61 r (21/05/02 13:30) MIN 313/023622 : 451 : MIN 313/023622 Operator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 67r (21/05/02 12:08) -0 30--IS 20-1 Mg 10- J 10 11"' I-- i* 15 20 Energy (keV) : 452 : Operator; Plymouth EM Unit C lient: none Jo b : WIN 313 Recovery 66r (21/05/0212:14) MIN 313/023622 Energy (keV) : 453 : Operator; Plymouth EM Unit C lient: none jo b : MIN 313 Recovery 70r (21/05/02 12:05) MIN 313/023622 Energy (keV) : 454 : Operator: Plymouth EM Unit CRent: none J o b : MIN 313 Recovery cp 71r (21/05/02 12:00) MIN 313/023622 : 455 MIN 313/023622 Operator: Plymouth EM Unit C lient: none Job : MIN 313 Recovery 74r (21/05/02 11:56) Mg 20- 15- 10-- l n ---------------------1 r- ' ------ 1 .............. . . l----------1--------- 1----------1----------1----------i-------------- y---- -1. i . r - 10 15 r-T -T -r 20 Energy (keV) : 456 : O perator: Plymouth EM Unit C lient: none J o b : MIN 313 Recovery 75r (21/05/02 11:52) MIN 313/023622 : 457 : Plymouth Electron Microscope Unit MIN 313/023622 Scanning Electron Microscope Examination and X-Ray Microanalysis. Study Number: MIN313 Huntingdon Life Sciences HCuamntbinsgdon PE284HS PUlnyimveorustihtyEolefcPtlryomn Mouitchroscope Unit DPlryamkeouCtihrcus PL4 8AA 01752 233092 : 458 : MIN 313/023622 Materials and Methods 1. SEM examination of urinary bladder tissue samples: Subsamples (approximately 0.5 x 0.5cm) were cut from fixed tissue samples (supplied in 70% ethanol) and critical point dried (protocol attached), Diy samples were mounted on specimen stubs with colloidal silver adhesive, sputter coated with gold and examined in a JEOL 5600 SEM at 15kV. Four images, each at 500x magnification, were collected to illustrate typical epithelial morphology. 2. SEM/X-Ray microanalysis of urine samples: Dry samples as supplied were carbon coated and examined in a JEOL 6100 SEM at 15kV. X-ray data were collected and analysed with an Oxford Instruments ISIS X-Ray Analysis system. Liquid samples as supplied were centrifuged, a 30pl droplet from each sample was placed on a carbon stub, air dried, carbon coated and examined as described for the supplied dry samples. 459 : Results 1. SEM Examination ofUrinary Bladder Epithelium. MIN 313/023622 : 460 : MIN 313/023622 : 461 : MIN 313/023622 : 462 : MIN 313/023622 50pm : 463 : MIN 313/023622 : 464 : MIN 313/023622 : 465 : MIN 313/023622 : 466 : MIN 313/023622 : 467 : MIN 313/023622 : 468 : MIN 313/023622 : 469 : MIN 313/023622 : 470 : MFN 313/023622 2. SEM/X-Ray analysis of Urine samples Representative images are provided from the supplied dried samples to illustrate typical crystal structure and deposit morphology. X-Ray analysis spectra from the supplied dried samples (labelled MIN 313), to show general elemental composition, and composition of specific crystals and deposits (as labelled) are provided. Similarly, spectra for the supplied liquid samples are provided (labelled MEN 313a). : 471 : MIN 313/023622 : 472 : MIN 313/023622 : 473 : MIN 313/023622 : 474 : MIN 313/023622 Specimen Preparation for Scanning Electron Microscopy Critical Point Drying (CPD) 1. Fix freshly excised sample appropriately (eg. 2.5% Glutaraldehyde in O.IM phosphate buffer) 2. Wash in buffer - 3x 15 minutes 3. Dehydrate in ethanol series 30% -15 minutes 7500%% --1155 mmiinnuutteess 9100%0%- 1- 515mminiuntuetses 100% -15 minutes 4. Fill CPD chamber with 100%ethanol and cool to 10C or less 5. Place specimens in CPD chamber 6. Close and flood with liquid carbon dioxide 7. Exhaust alcohol from chamber. 8. Leave sample 30 minutes in chamber to infiltrate with carbon dioxide. 9. Exhaust remaining ethanol 10. Heat chamber to critical point 11. Vent carbon dioxide gas from chamber 12. Remove sample from chamber and mount for SEM examination. : 475 : Operator : Plymouth EM Unit C lient : andrew lawrence HLS Job : MIN/313 control (05/03/02 12:33) MIN 313/023622 476 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 81 centre deposit (27/02/02 18:58) MIN 313/023622 : 477 : Operator : Plymouth EM Unit C lient : andrew lawrence HLS Jot) : MIN/313 82 crystal (27/02/02 19:06) MIN 313/023622 : 478 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 82 background (27/02/02 19:09) MIN 313/023622 : 479 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 83 crystals (27/02/02 19:12) MIN 313/023622 : 480 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 84 crystal (27/02/02 19:15) MIN 313/023622 : 481 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 85 crystal (27/02/02 19:18) MIN 313/023622 : 482 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 86 crystal (27/02/02 19:20) MIN 313/023622 : 483 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 87 crystal (27/02/02 19:54) MIN 313/023622 : 484 : MIN 313/023622 Operator : Plymouth EM Unit Client : andrew lawrence HLS Job ; MIN/313 87 background (27/02/02 19:57) 80- 60--I : 40- 0 N C 20-- -- I-------- 1-------- 1-------- 1-------- I-------- 1-------- !-------- 1 - 10 20 Energy (keV) : 485 : O perator: Plymouth EM Unit C lie n t: andrew lawrence H IS J o b : MIN/313 88 crystal (27/02/02 20:02) MIN 313/023622 : 486 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 89 background (27/02/02 20:07) MIN 313/023622 : 487 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 89 crystals (27/02/02 20:09) MIN 313/023622 : 488 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 90 (27/02/02 20:13) MIN 313/023622 : 489 : O perator: Plymouth EM Unit C lie n t: andrew iawrence HLS J o b : MIN/313 92 crystal (27/02/02 20:18) MIN 313/023622 : 490 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 93 crystal (27/02/02 20:25) MIN 313/023622 : 491 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 93 background (27/02/02 20:28) MIN 313/023622 : 492 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 94 crystal (27/02/02 20:33) MIN 313/023622 : 493 : Operator : Plymouth EM Unit Client : andrew lawrence HLS Job : MIN/313 94 plates (27/02/02 20:38) MIN 313/023622 Energy (keV20) : 494 : O perator: Plymouth EM Unit C lie n t: andrew lawrence H IS J o b : MIN/313 95 crystal (27/02/02 20:43) MIN 313/023622 : 495 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 96 cast (05/03/02 12:03) MIN 313/023622 : 496 : Operator : Plymouth EM Unit C lient : andrew lawrence HLS Job : MIN/313 97 cast (05/03/02 12:06) MIN 313/023622 : 497 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 98 cast (05/03/02 12:11) MIN 313/023622 : 498 : O perator: Plymouth EM Unit C lie n t: andrew lawrence HLS J o b : MIN/313 99 cast (05/03/02 12:20) MIN 313/023622 Energy (keV) : 499 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313A 82A crystal (11/03/02 13:44) MIN 313/023622 : 500 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313 A 85A (11/03/02 14:00) MIN 313/023622 : 501 : O perator: Plymouth EM Unit C lie n t: andrew lawnence Jo b : MIN 313 A 86A (11/03/02 14:25) MIN 313/023622 : 502 : O perator: Plymouth EM Unit C lie n t: andrew lawrence J o b : MIN 313 A 87A (11/03/02 14:32) MIN 313/023622 : 503 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313 A 88A (11/03/02 14:30) 50-- 40-- 30- MIN 313/023622 5 8 -4 15 Energy (k : 504 : O perator: Plymouth EM Unit C lie n t: andrew lawrence J o b : MIN 313 A 89A (11/03/02 14:41) MIN 313/023622 : 505 : O perator: Plymouth EM Unit C lie n t: andrew lawrence J o b : MIN 3 1 3 A 90A (11/03/02 14:49) MIN 313/023622 : 506 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313 A 91A (13/03/02 09:33) MIN 313/023622 : 507 : O perator: Plymouth EM Unit C lie n t: andrew iawrence Jo b : MIN 3 1 3 A 92A (13/03/02 09:35) MIN 313/023622 : 508 : O perator: Plymouth EM Unit C lie n t: andrew lawrence J o b : MIN 313 A 93a (14/03/02 09:09) MIN 313/023622 : 509 : O perator: Plymouth EM Unit C lie n t: andrew iawrence Jo b : MIN 313 A 94a (14/03/02 09:13) MIN 313/023622 : 510 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : M IN 313A 95a (14/03/02 09:15) MIN 313/023622 : 511 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 3 1 3 A 96a (14/03/02 09:19) MIN 313/023622 i 15 : 512 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313 A 97a (14/03/02 09:24) MIN 313/023622 : 513 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Job: M IN 313A 98a (14/03/02 09:27) MIN 313/023622 : 514 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313 A 99a (14/03/02 09:31) MIN 313/023622 : 515 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313 A 99a crystals (14/03/02 09:35) MIN 313/023622 : 516 : O perator: Plymouth EM Unit C lie n t: andrew lawrence Jo b : MIN 313 A 100a (14/03/02 09:41) MIN 313/023622 : 517 : ANALYTICAL PHASE REPORT MIN 313/023622 ANALYTICAL PHASE REPORT STUDY TITLE APedrmfliunoirsotroactitoannetsoulCfoDnyRlaFtslufoorrid1e3 (WPOeeSkFs;FTo-l7lo6w61e.d4)bTyoax4ic-WityeSektuRdyecboyvIenrhyaPlaetriioond DATA REQUIREMENTS Analytical Method Requirements STUDY DIRECTOR HuntingTtoenrrLyiJf.eKSecniennyces, Ltd. ANALYTICAL PHASE COMPLETED ON March 10,2004 PERFORMING LABORATORY StP3ah0tEeo5xn8CyeoR:glee8lens1ge4Rae-r,e2csP7heA2aD-r1c1r0hi6v38e901 STUDY SPONSOR 3SMt.BPCuaoiulrldp,ioMnrgaNt2e25T05-o12x3Ei3c--o032l2o2g0y PROJECT ExPyrgoetnocSotluNdyumNbuemr.bMerIN02/331-0376 Total Pages: 217 : 518 : MIN 313/023622 Exygen Study No.: 023-076 GOOD LABORATORYPRACTICE COMPLIANCE STATEMENT Exgyen Study Number 023-076, the analytical phase of the study entitled "AcoPdnemdrfuilnuciotsertdoraotfciootarnn3etsMoulCfCoDnoyrRpl oarFtasltuefoorTrido1ex3ic(oWPloQegeSykF,s;wFTao-sl7lo6pw6er1ef.do4r)bmyTedoaxi4inc-iWtcyoemeSkptulRidaeynccoebvyewryiItnhPheaOrlaEiotCidoD,n" PErxiyngceipnleRseosefaGrcoho.d Laboratory Practice (as revised in 1997), ENV/MC/CHEM(98)17 by PErxiyngceipnaRl Iensveeasrctihgator Huntingdon Life Sciences Ltd JS3oMphonnCsBoourrptReonreahptoreefTsfeonxtiactoivloegy Exygen Research : 519 : Date Date Date Page 2 of 217 MIN 313/023622 Exygen Study No.: 023-076 QUALITYASSURANCE STATEMENT E7an6xa6yl1gy.et4inc)aRTl eospxehiacaristceyh'SostfuQdutyhaelbiytystIuAndhsyasulaertaninoticnteleAUdd,nmi`tTinreeirsvftilreuawotiroeodnoEctotxayCngeDesnuRlSfaottunsdyfylorNF1luu3moWrbiederee0k2(sP3FO-0oS7llF6o;,wtTehdebacycoard4in-Wg teoekExRygeceonvRereyseaPrecrhio'sd"S.tanAdallrdreOvpieerwaetidngpPhraosceesduwreerse, thinespSetucdteydPfroortoccooln,dauncdt aSltludayppDliicraebctleorGanododtoLmaabnoaragteomryenPt.ractice Standards. All findings were reported to the Phase InsDpeacteted DMtoataEenxRaayngegedpemonretPneItd 1. AIEnxnsttarrlauycmstiiesonnbt,yation 10/07,08/02 11/22/02 2. Raw Data Review 121/10/227,0,239/0,2 03/17/03 3. Raw Data Review 11/27,12/02-05/02 1122//1129//0022, 4. Raw Data Review 03/14,17/03 0077//1271//0033, 5. RDerapfotrAt Rnaelvyiteiwcal 10/29-31/03 1013//1110//0034, 6. RFienpaolrAt Rnaelvyiteiwcal 03/08/04 03/08/04, 03/10/04 SDtuaMTdteyeasRnDtaeFierpaeeoccmritltoeeitnrdytatnod 12/09/02 03/10/04 04/16/03 03/10/04 03/10/04 03/10/04 / 7 w **' / K MQuiwalaityNAabsestuarnaince Auditor fatibcL f y Date Exygen Research Page 3 of 217 : 520 : MIN 313/023622 Exygen Study No.: 023-076 CERTIFICATION OFAUTHENTICITY Tthheisrarwepdoartta, ffoorrEthxeysgteundyS.tudy Number 023-076, is a true and complete representation of Submitted by: E3S0xta5yt8geeRCneoRslleeeasgreeca,hrPcDhAriv16e801 (814)272-1039 Principal Investigator, Exygen: EPrxeysgideennRt esearch /t-nsJrt..oy Date Exygen Research Facility Management: ^ 6 h n M .EFla'he--rty ' EXV/x,i/cy,eagePD,nr-enRsriedseenatrch Date Huntingdon Life Sciences, Ltd Sponsor Representative, 3M; J3oMhnCBorupteonrahtoefTf oxicology Exygen Research Date Page 4 of 217 MIN 313/023622 Exygen Study No.: 023-076 STUDY IDENTIFICATION APedrmfliunoirsotroactitoannetsouClfoDnyRlaFtslufoorrid1e3 (WPOeeSkFs;FTo-l7lo6w61e.d4)bTyoax4ic-WityeSektuRdyecboyvIenrhyaPlaetriioond PROTOCOL NUMBER: MIN/313 EXYGEN STUDY NUMBER: 023-076 TYPE OF STUDY: Analytical SAMPLE MATRIX: TEST SUBSTANCES: Rat Liver, Serum, and Urine PPPeeernrfftllauudooerrcooaoofccluttaaonnreeossouuclltffaoonnnoyaitlceFA(lPucFoidrOid(SPe)F(aOPnOAd S) F), SPONSOR: STUDY DIRECTOR: 3BSMtu.iPlCdaiounlrg,pMo2r2aN0te-52T5Eo1-x30i23c-o3l2o2g0y Terry J. Kenny WHAHPCEauluocmnn2oott8blniilnn4rbeiggHuydddrgSRooyennoshaLdirifee, Sciences England Ltd SPONSOR REPRESENTATIVE: John Butenhoff B3SMLuiPlCdaiounrlg,pMo2r2aN0te-52T5Eo1-x03i23c-o3l2o2g0y PERFORMING LABORATORY: E3S0txa5yt8geeCRneoRslleeeasgreeca,hrPcDhAriv16e801 TAINMAELTYATBILCEA:L PHASE ESEAtxxnuppadeelyyrriitImminciaeetlnniatPtataihlloaTSnsteeDarrCmattoDienm:aatpteilo:entioDnaDtea:te: 00119320////21026009////00001242 Exygen Research Page 5 of 217 : 522 : MIN 313/023622 Exygen Study No.: 023-076 PROJECT PERSONNEL TThhee SfotulldoywDinigrepcetorsrofnonretlhifsropmrojEexcyt gweansRTeesreryarJc.hKWenerneyaastsoHcuiantteindg,wdoitnhlvifaerioSucsiepnhcaesseLstod.f the study: Name Richard A. Grazzini Emily Decker Paul Connolly Karen Risha Lawrence Ord Rickey Kelley Chas Simons Carisa Kelley Joe Gallagher Chris Pfleegor Xiaoming Zhu Title President Scientist Technical Lead-LC/MS Scientist Sample Custodian Sample Custodian Technical Lead-GC/MS Technician Scientist Scientist Technician Exygen Research : 523 : Page 6 of 217 MIN 313/023622 Exygen Study No.: 023-076 TABLE OF CONTENTS Page TGIOTOLEDPLAAGBEO.R...A...T...O...R...Y...P...R...A...C..T...I.C...E....C...O...M...P...L..I.A...N...C...E....S..T...A..TEMENT.............................. 2 QUALITY ASSURANCE STATEMENT.......................................................... PTLCSLRTAIIESSUORBTTDTJLOOEIYEFCFFOIITCTFDFIAAPEGCETBNUORILTORSNEINOFESTISN.OEC...N..NFA......ETAT....LS..IU..O........T....N.....H..............E.............N...............T............I........C.............I........T............Y............................................................................................................................................................................................-...............................................................................................................................................................................................................................8459 L2431....I0000STITOSNOUEBTSFMJRTEAMOCSPTDAYPIURSEVTCNYEETD....MI....IO..C.....N...E..........S............................................................................................................................................................................................................................................................................................................................................................................................................................................................................................1111111022 56..00666666.....311254RD...121IECEnPFhSPPDsrrEtCFOeeroORpusRSmcamESFIrraPiNaetEapTotnCintxgtIoidOotErSnranPaNepMocFnohftOOsAifySoiL.FtATt.ni.aCv.EAn.PE.i/R.dtMNrx.y.oaI.tA.A.rSrc.d..ae/L..L.sc.Md..Yt...au..i.S...oTn..r....ned.I..I...C.n....PF....s...A.or...t....or..r..L.u...t.c...i.m...Me.f......i.d...e.c....E.un..a......rt.Tt.....i.e....ao.H.......n..n......O....d.....S........DO...o...........l.p........u......e.......t..r.....i.....a..o.........t......n.i.......n.....s.......g.....................C.....................o...............n................d.................i.......t........i........o..................n...............s.........................................................................................................................................................................................................1111111135325454 66..65..12 DPFeOscAripatniodnPoFfOGSCQ/MuaSntIintasttriounmaenndt aEnxdaOmppeleraCtianlgcuClaotniodnit.i.o..n..s...............................................................1167 7891...00006.0.6ERC.R2XEOEPSPNTOUECERLSLNFITUMTSQSI.E.OuI.N.Oa.N.nT.N.t.AOi.St.a.LF...t..i.DD..o....nE.A.....Sa.T...n.I.A..Gd......NEA.....x.N.....a....D..m..........p.S.....l..A..e.......M.C.......a..P..l..Lc....u..E..l...aS...t....i...o......n...................................................................................................................................................................................................2222110180 1 3 67 Exygen Research : 524 : Page 7 of 217 MIN 313/023622 Exygen Study No.: 023-076 LIST OFTABLES Page Table L Summary ofPFOS in Control Rat liver Samples....................................................23' Table IL Summary of PFOS in Control Rat Serum Samples ........................................ 23 Table HL Summary of PFOS in Control Rat Urine Samples...........................................23 Table IV. Summary ofPFOA in Control Rat Liver Samples..................,*....... 24 Table V. Summary of PFOA in Control Rat Serum Samples.........................................24 Table VL Summary of PFOA in Control Rat Urine Samples..........................................24 Table VC. Summary of POSF in Rat Liver Control Samples........................................... 25 Table VCL Summary of POSF in Rat Serum Control Samples......................................... 25 Table DC Summary of POSF in Rat Urine Control Samples........................................... 25 Table X. Summary of PFOS Fortification Recoveries in Rat liv e r...............................26 Table XL Summary of PFOS Fortification Recoveries in Rat Serum............................. 27 Table XU Summary of PFOS Fortification Recoveries in Rat Urine...............................28 Table XCL Summary of PFOA Fortification Recoveries in Rat Liver...............................29 Table XTV. Summary of PFOA Fortification Recoveries in Rat Serum............................ 30 Table XV. Summary of PFOA Fortification Recoveries in Rat Urine..............................31 Table XVI. Summary of POSF Fortification Recoveries in Rat Liver...............................32 Table XVIL Summary of POSF Fortification Recoveries in Rat Serum............................33 Table XVHL Summary of POSF Fortification Recoveries in Rat Urine............................34 Table XIX. Summary of PFOS Residues in Rat Liver Samples.........................................35 Table XX. Summary of PFOA Residues in Rat Liver Samples.........................................40 Table XXL Summary of POSF Residues in Rat Liver Samples.........................................45 Table XXC. Summary of PFOS Residues in Rat Serum Samples....................................50 Table XXCL Summary of PFOA Residues in Rat Serum Samples............................... 55 Table XXTV. Summary of POSF Residues in Rat Serum Samples...................................60 Table XXV. Summary of PFOS Residues in Rat Urine Samples..................................... 65 Table XXVI. Summary of PFOA Residues in Rat Urine Samples................................... 69 Table XXVII. Summary of POSF Residues in Rat Urine Samples...................................73 Exygen Research Page 8 of217 : 525 : MIN 313/023622 Exygen Study No.: 023-076 LIST OF FIGURES Figure 1. Typical Calibration Curve for PFOS.................. 78 ' Figure 2. Typical Calibration Curve for PFOA.............................. 79 Figure 3. Typical Calibration Curve for POSF.................- .............................................80 Figure 4. Chromatogram Representing a 0.1 ng/mL standard for PFOS and PFOA......81 Figure 5. Chromatogram Representing a 25 pg/mL standard for POSF..........................82 Figure 6. PCFhOroAma(EtoxgyrgaemnRIDep0r2es0e1n6t8in4gCaoCntornotlrAol,RSaett:L1iv0e0r8S02amB)p..l.e...f.o..r...P..F..O...S....a..n..d.......83 Figure 7. C(EhxryogmeantoIDgr:a0m20R2e8p7re7s,eSnetti:n1g0a2C50o2nAtro).l..R...a..t..L...i.v..e..r..S..a..m....p..l.e...f.o..r...P..O...S...F..............84 Figure 8. PCFhOroAm,a(tEogxryagmenRIeDp:re0s2e0n1t6in8g2aCConotnrtorloAl R, aSteSt:e0ru9m25S0a2mAp).l.e..f..o..r..P...F..O...S....a..n..d.....85 Figure 9. C(EhxryogmeantoIDgr:a0m20R1e6p8re2sCentrtli,nSgeat:C0o9n2t6ro0l2RBa).t..S..e..n..i..m....S..a..m...p..l..e..f..o..r..P...O...S..F..,...........86 figure 10. PCFhOroAm,a(tEogxryagmenRIeDp:re0s2e0n1t6in8g3aCConotnrtorlolAR, aSteUt:r1in0e0S30am2Apl)e...f.o...r..P..F...O...S...a..n..d.......87 Figure 11. C(EhxryogmeantoIDgr:a0m20R2e8p7r5esCentrtli,nSgeat:C1o0n1t1ro0l2RAa)t..U...r.i..n..e..S...a..m...p..l.e...f..o..r..P..O...S...F..,............88 figure 12. CPFhOroSmaantodgPraFmOARe(pErxeysegnetninIDg :C0o2n0tr1o6l8R4aStpLkivAe,r Sfoertt:if1ie0d08w0i2thB)1.0...n..g../..g...o..f..... 89 Figure 13. CPOhrSoFm(aEtoxgyrgaemn IRDe:p0re2s0e2n8ti7n7gSCpoknAtroSleRt:a1t L02iv5e0r2fAor)t.i..f.i.e..d...a..t..5...p..g.../.g...w...i.t.h.........90 Figure 14. oCfhProFmOaStoagnrdamPFROeApre(Esexnytgineng CIDo:n0tr2o0l1R6a8t2SSeprukmAf,oSretitf:i0ed10w0i2th021A0)n..g../.m...L......91 Figure 15. Chromatogram Representing Control Rat Serum fortified with 5 pg/mL of POSF (Exygen ID: 0201682 Spk A, Set 092602B)...................................92 figure 16. Chromatogram Representing Control Rat Urine fortified with 10 ng/mL of PFOS and PFOA (Exygen ID: 0201683 Spk A, Set 100302A)................93 figure 17. CPOhrSoFm(aEtoxgyrgaemn IRDe:p0re2s0e2n8ti7n5gSCpoknAtro, lSReat t1U0r1i1n0e2fAor)t.i.f..i.e..d...a..t..5...p..g../..m...L....w...i.t.h.... 94 Figure 18. C(EhxryogmeantoIDgr:a0m20R3e5p5r3es,eSnptoinngsoRraItDL:i9v1erSSeat:m1p0le21fo0r2PAF).O...S...a..n..d...P...F..O...A............. 95 Exygen Research Page 9 of 217 : 526 : MIN 313/023622 Exygen Study No.: 023-076 Figure 19. Chromatogram Representing Rat Liver Sample for POSF (Exygen ID: 0203515, Sponsor ID: 52, Set: 102502A)........................................................96 Figure 20. Chromatogram Representing Rat Serum Sample for PFOS and PFOA S(Eext:y1g0en02ID02:A0R20).3..4..5..0..,..S...p..o..n..s.o...r..I.D...:..8..6...M...-..G...R..O...U...P.'..4...W....e..e..k...4..,.........................97 Figure 21. C02h0r3om37a7t,ogSrpaomnsRoerpIDre:se1nlMtin-gGRroaut pSe3ruDmayS9a2m,pSleet:fo0r9P2O60S2FB()E...x..y..g..e..n...I.D...:...... 98 Figure 22. C(EhxryogmeantoIDgr:a0m20R3e5p7r3es,eSnptoinngsoRraItDU: r1in1eSSeta:m1p0l0e3f0o2rAPRFO)..S...a..n..d....P..F..O...A.............99 Figure 23. C02h0r3o5m7a3t,ogSrpaomnsRoerpIDre:se1n1t,inSgetR: a1t0U11ri0n2eAS)a..m...p..l.e...f..o..r..P..O...S...F...(..E..x..y..g...e.n....I.D...:......100 LIST OFAPPENDICES Page Appendix A AStmudeyndPmroetnotcsoalnMdIDNe/3v1ia3ti(oEnxsy..g..e..n...S..t..u..d..y...N...o.....0..2..3..-..0..7..6..)..a..n..d...................... 101 Appendix B PAenrafllyutoicroahl eMxaentheosdu:lfMoneattheo(dPoFfHASn),aPlyesrifslufoorrotohectDaneetseurmlfoinnaattieo(nPoFfOS) and P(EexnytagdeencMafleutohroodoNctoan. EoixcMA-c0i2d3(-P0F7O1,AR)eivnsRioant l1iv)e...r.,...S..e..r.u..m.....a.n...d...U...r.i.n..e...139 AP(EenxrafyllyugtoeinrcoaMol cMetatehntoehdsoudNl:fooM.nEyexlthMFoldu-0oo2rfi3dA-e0n7(aP1lyAOs,SisRFf)eovirnistRhioeantD2Ue)rt.ei.n.r.me..,..iS.n..ea..rt.iu.o.m.n...o.a..nf..d..L...i.v..e1r83 Exygen Research : 527 : Page 10 of 217 MIN 313/023622 Exygen Study No.: 023-076 1.0 SUMMARY EdextyegrmeninRateiosnarcohf epxetrraflcuteodrooacntdanaensuallyfozneadtera(tPlFivOeSr,) saenrudmp,eanntaddeucraifnleuosraomocptlaensoifcoratchide (dPeFteOrmAi)naaticocnorodfinpgerftlouoEroxoycgtaenncsMulfeothnoydl flEuxoMrid-0e2(3P-O07S1F) (aAccpoprednindgixtoBE)xyagnedn Mfoerthtohde ExM-023-071A (Appendix C). Tspveaghrl/euimdmlaiLmtiaofinotndrofrusaqtrutiundsaeiene.rtsuitTmapthieaeornnfldoifmroumirrtiePndoFef.OaqStTuahaEnnextdiLytPagOtFeiQoOnnAfo(fEoirnxreyrPaagcOtehnlSimvFeSarittnurwidxraiasewts1:lai0sv0ned2gre3/wtg-e0aram7sn4di2n.e15ad0npdnigng//g0am2am3Ln-e0idnt7h25ro.a)5dt. Hsswusaaesohmmrewuidcppmehwllveeiwhensariacic,lzsrihqteehd,uaienesootcenteLdrlrefOymotahQ0rsieen.s1dLefedorOtmuhruQmeLPsLiFwfnaoOOgnarQsAd0e.uafm1acsonehimrnddPmeLfOPoaasFtSranrOiFmsdxeSiprnb1uli.yemn0seaa,egrlfu0iaaqsmc.c2uahtmobo-mtyrpa1olaane.tf0rdfiw1amxic0netLwioogsrraofhosolmterfsdfeu2soe.r.crtieanTlrsiemevh,seeiarnoLn.endOdlDy0Qu.u01sfe.i0ong5trgofPmoaOlriLm1SliFwvimteeaiLrdns, PtdddsPhaeFeeFemtttOOeeerpcccSaAltttteeeeislidddnni.vitllnehteePhvrveFaeeesnrOlalrayssmaSttottlpoioifunvlret1ehi1tsnr2he5er,es81ara,asn0r0maat0g0mtlpe0inudlvpegrenlsif/erngmrsroe/samLmarnsam.Lgannmep.gTodlepenhdPfls-eer.FdofrsreeOmPorteaAwFmcnnOatogiensSnnedod-nindtlonhe-ferdvettPoheeeOmcrteleastSrectntadtFooesteldnsde8re-evu2ldretem7ueevl0mctseetlsncetsasogtdamet/5modgipn3.pll15eela8T,ves0n,seh60yrle0ra0saro0netnnfgonggwetg4eh/dga/d7em.s1ffrrLrn0Paooo.FtmnmOsPgTe/OnnAhmrooueSLnnimrFne--. was no POSF detected in any of the rat urine samples. The average percent recoveries standard deviations for PFOS in rat liver, serum, and urine samples were 96% 11%, 102% 14%, and 87% 11%, respectively. The average percent recoveries standard deviations for PFOA in rat liver, serum, and urine spwaeemrrcepeln9et9s%rewceorve1e8r9%i9e,%s92%st9a%nd1,a8r1%d05,d%aenvdia9ti91o4%n%s,foa1rn8Pd%O,1Sr1eF0s%pinecrtaivt1e7lli%yv.e,r,ressepreucmti,vaenlyd. urTinhee saavmerpalgees 2.0 OBJECTIVE T(sPphOeecSoimFb)je,encpsteiorvffeluroaoftrotlhiovicsetrpaanhneadssueslweforaunsmattoesa(dmPeFtpeOlremSs)ia,ncaecnlodervdpeienlnsgtoatdfoepPcerarofftllouucooorrool ooMccttIaaNnn/eo3si1cu3lafc(oAindyp(lpPfeFlunOodrAiixd)eAin). Exygen Research Page 11 of 217 : 528 : MIN 313/023622 Exygen Study No.: 023-076 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFOS and PFOA in rtramFhaenlteseudutloDhilPvrtoeiseeddtnrefeotreaam(rnndPtdtieOihntcsleaSaeetfFddriloue,)unmtoi`enMrorosmRfaeoPmaitcnhtetpaaoUrltfndeilroousioinn,ocfeuroo,AsAfhiSncPneeigxaOrdulatyShmn(sPeFei,FssaiauOnnfnlaofAdrolray)LntttaiihilncvtieeaveRleDr(PmrabeFtayteenHLtGrhdiSmvoCs)edi,e/nrMrPe,aunetSSmitroe.if"trlnlsuueaomdomr,fop"aPolMneecdsrtea,ftUlnuhuesrooisidnrnuoegloof"fotchntaAaenanntdaeeansla(yauPllsslyFfiosotOintcfhSoyae)rll TnthuhemeabsnteaurldyMyticIwaNla/es3x1ip3ne.itriiamTtheedentoaanlnatelDyrmeticcieanmlatebixoepnre6dr,aimt2ee0wn0t1aa,sl wOsthcaetrotnbdteahrtee29swt,u2ad0sy0S2de.iprteecmtobresri2g0n,ed20p0r2o,toacnodl 4.0 TEST SYSTEM Trehceeicvoendtrofrlorzaetnlivoenr adnryd siecreumonusJeudnefor1t2h,e2m00at2rixanbdlanAkusgaunsdt m7,at2ri0x0f2orftrifoimcatiPoenls-Fwreeerze Biologicals, Rogers, Arkansas. Nr2e0icn0ee2ivtyae-ndedifglrohogtzgereandtoilnnivbedryr,yEnixicnyeegtaeytn-EepixegyrhsgtoenrnantefrlsoeamrnudmHp,ulanactnienddgsidneovfneronLztiyef-nenisSntcoeirearngactee.ms ionneSseapmtepmlebsewr e1r0e, Sasasmocpilaeteldogw-iinthatnhdiscshtauidnyo. fSctuosrtaogdeyriencfoorrdmsawtiiolnl bceankebpet faotuEnxdyignenthReersaewarcdhataanpdacaktarguee cstoupdyy.of the storage records can be found in the raw data package associated with this 5.0 REFERENCE MATERIAL The analytical standard PFOA was received at Exygen on July 3, 2002 from SigmaAManlaadylryict1ih5c,aal2n0sdt0as0ntodfrraeordmdrPe3fOMriSgEFernawvteiardso.nremTcheeenivtaeanldaTlayettciEhcxanlyogsloteagnnydoaanrndJduPSnFeeOrv6Si,cw2es0a0sa2nredfcrseotimovreedSdiagftrmoEazx-eyAng.lednTriohchne and stored ambient The available information for the reference material is listed below. CoPPPmFFOpOOSoASFund ExygenSSSPPPIn112v134e829n78tory No.. 1T5SC7DNR20A1N1H8oQ. Pur998it986y...969(%) Exp000i876ra///300ti136o///n000633Date Exygen Research Page 12 of217 : 529 : MIN 313/023622 Exygeu Study No.: 023-076 The molecular structures of test substances are given below: NaCmhee:mPFicOalSName: Perfluorooctane sulfonate Molecular Weight: 499, as shown NaCmhee:mPFicOalAName: Perfluorooctanoic acid Molecular Weight: 413, as shown NaCmhee:mPOicaSlFName: Perfluorooctanesulfonyl fluoride Molecular Weight: 502, as shown 6.0 DESCRIPTION OFANALYTICALMETHOD The analytical methods ExM-023-071 and ExM-023-071A were used for this study. The mdariefefteihnroecdnlustdsweademrweplivethesrtiszhieiossnraeendpdoartfott.emr athkee manianloyrsitsypoofgtrhaephsiacmalpcloesrrewcetiroensc.omTphleerteedvitsoedamlloewthofdosr 6.1.1 PFOS and PFOA Extraction Procedure Aaliq1u00otpoLf aulriiqnueo,taonfdth0e.1segruomf tshaemlpivleer(1(.10.0mgLffoorr llaabboorraattoorryy ccoonnttrroollss)), w20e0repuLsetod 1fo.0rmthLe Exygen Research Page 13 of 217 : 530 : MIN 313/023622 Exygen Study No.: 023-076 extraction procedure. After fortification of appropriate samples, the serum and urine sumaathaldpiimedqnteuupsoduotleept1ase0naowrndfnmdeaotrhLtnteaheenbwestmrailowtimihvulalgepsiTrhllieytdtssepuaerewmcpaweIptnoralWteees2sdsa0thwrtoaeamennrkr.LtseeofnewTharfohriotecmehrdo-osTnfegdry2reoipu0tnmiemiomzIneeiaWeandndcudwahtSetiuesFstarrh.EimanTanecphdotliseelstau'hssmamueanempnldmil.vep5iselzeTrmewshsrwLeaefmrnoeeorrptfevhl-eaeocscie1stenweatmtoxmerneiirfpnediutlurbegfitsreoeleo.drwuw-agAenahrnde1st eluted with 2 mL of methanol. Each sample was analyzed by LC/MS/MS electrospray. 6.1.2 POSF Extraction Procedure Aext5r0a-c1ti0o0nppLroacleidquuroet.oAf tfhteersfeorrutmificaantdiounrionfeaapnpdro0p.1riagteofsathmepllievse,rthweesreamuspeledsfworetrheeheated to 60C-80C for - 3-4 minutes and then analyzed by GC/MS. 6.2 Preparation ofStandards and Fortification Solutions Asolsuttoiockn sotfaPndFaOrdS swoalustipornepoafrePdFoOnAAwugaussptr2e,p2a0re0d2oans sJpuelycif8i,ed20i0n2EaxnydgeansmtoectkhosdtaEndxaMrdF0dm2irsLo3sm-oo0lf7vteh1inai.scghTs1ohs0lteuomtscitkoognac,oknadfsteb1aar.n0icndhpgagrisnd/tmgasnotLdhlauefrtovdiorotn(ilfcusiomcwrareeteiruocetpnepdtsroetafp1ona0rdr0eapdrmudraListtoyawluciattonhinodmcnesewantlhtatrasacntpoioonrlne.teponaftr)e1di0n0bympgtea/tmkhiaLnngobly1. fAor0t.i1ficpagti/omnLsftaonrtdiafircdaatinodnbsrtainngdianrgdtwheasvporleupmaereudpbtyo t1a0k0inmg L10wmithLmofetthhaen1o.l0. pg/mL mixed AandsevtaorifosutsancadlairbdrsatcioonntsaoinluitnigonPsFiOn AtheanfodllPoFwOinSgwmaasnpnreerpared dilution of the 0.1 pg/mL Initial Cone. (ug/mL)1 0.1 0.1 000..00.10052 'o f PFOA0.0a0n1d PFOS Volum5e (mL) 11121000 Dilute1d0t0o (mL) 111110000000000 Final Cone. (ug/mL) 00000...0..0000000000015252 0.0001 A stock standard solution of POSF was prepared on June 11,2002 as specified in Exygen Fm215r9eo9mtmh0oLptdhogiEf/smxtshMLoel-usb0tty2oio3cndk-,0isa7asn1od1Al0vb.0irn0iTnggph-igen/gm0st.to2Lhceckgavslooitbaflunrtamdhtaieeordnustpsasottnaolduna5tdir0oadrnmd(wcLsooawrlsrueipttchiroteemnpdaewrftehoadsarnpapotrule.rapitcayor)endicnebnymtrteaattkhioiannngool-.f Exygen Research Page 14 of 217 : 531 : MIN 313/023622 Exygen Study No.: 023-076 A set of standards containing POSF was prepared by dilution of the 1000 pg/mL: Initial Co1n0e0.0(pg/mL)1 Volume (mL) 5 Diluted10to (mL) Final Cone. (pg/mL) 500 111000000000 201..55 . 1i1o00 - 21550000 'o f POSF11000000 0.25 0.1 1100 2150 The stock standard solution and all fortification and calibration standard solutions were sptroerpeadratiinonacarnefbriegeforautnodr in(4thera2wCda)tawahsesnocinaotetdinwituhseth. is rDepoocurtm. entation of standard 6.3 Chromatography QraenutyeannottfiiofthniceatictmioonentrooofflPPsFaFmOOpAAlewsanacdosr-PreF8sOp.6oSnawdnidansg- at8occ.8tohmme painlniasfhloyertdePrbFeyOteLSn.Ctio/PMneSatik/mMsewSs.eerleecntorotsdpertaeyc.teTdhine -Qanu3aal.yn5tteimfircienate.tinotnPioeonafktiPsmOweSeoFrfewPnOaosStaFdc.cetoemctpeldishinedthbey cGoGnt/rMolS.livTehresaremtepnletiocnortriemsepoonfdPinOgStFowthaes 6.4 Instrument Sensitivity Tcohnecensmtraatliloenstofs0ta.0n0d0ar1dpga/mmoLuonft PiFnOjeActeadnd dPuFrOinSgantdhe25cphgr/ommLatfoogrraPpOhSicF. run had a 6.5.1 Description of LC/MS/MS Instrument and Operating Conditions Instrument: PE SC1EX API 365 Biomolecular Mass Analyzer Interface: SCIEX Turboion Spray Liquid Introduction Interface Computer. Dell OptiPlex GX110 HSoPfLtwCa:re: PHEewSlceitetxHHPAaPPnckaVQlayaurcsdatut(PvuHemurPms)DipoSenegra1ise.1sse1r100 Exygen Research Page 15 of 217 : 532 : MIN 313/023622 Exygen Study No.: 023-076 HHPP ACoultuomsanmOplveern HPLC Column:Genesis Cg (Jones Chromatography), 2.1 mm x 50 mm, 4p CIMMFnlojoooelwbbuciimtlliReeonanPPtTehhVe:aamossleep.:.((:1AB05).)33::p5mL2MCImVetMmhainAn.oml monium Acetate.in. .W. ater Time 250..00 %90A 9100 %10B 9100 1994..05.0 1000 11900000 2104..05 9900 1100 Ions monitored: APnFaOlvAte neMgoatdieve Trans4it1i3on-M3o6n9itored ReteAntpiopnr8o.T6xiimmaet(emini PFOS negative 499 ^ 99 8.8 6.5.2 Description of GC/MS Instrument and Operating Conditions Instrument: HChewrolmetat-tPogarcakpahrd/mmooddeell56987930mSaesrsiesseGleacstive detector Column: Restek RTX-502.2,60 m x 0.25 mm ID, 1.4 pm df Oven Temperature: Hfoorld0 amti4n0.C for 4 min., ramp 20C/min. to 100C, hold Injector Temperature: 220C TCraarnrisefreGr Lasin:e Temperature: GC Head Pressure: Injection Mode: IInnjjeeccttiioonn LPuinregre: Delay: H42mr5S2e2pepmimm0nllsiiima.tiun,Clifemnr.oIsadDsrme0rsp.t2or-5fa8it0mghheiptnsag.in,/lmaarlasyimsnti.pctao8l03ru0pnsp.i/smi,ihno. ltdoa6t03p0spi,shi ofoldr 0.5 Exygen Research Page 16 of 217 : 533 : MIN 313/023622 Exygen Study No.: 023-076 PIESnucljearecgnctetrMiooFnnolodMVweo:utlloutimpSlpei:elirtVVoelntta:ge: F110rommmTLA/mTiUnNE +200V SIM MDwoneiltloTriemdeI:ons: ITRnoetettaegnlratriutoonnr:tTimime:es: mH7--375e/.zw5mm'slsmien6tiut9n-tPue(qstaecuskaanrtidtaCtihoenmioSnta)t;io1n31s-,o2ft1w9a(rqeualifier ions) 6.6.1 PFOA and PFOS Quantitation and Example Calculation Fifteen microliteis of sample or calibration standard were injected into the LC/MS/MS. Tlcionhneecaperenatrrekagtarireoesnasiwwoanas)sdmbeyteearsAmunrinaeldeydastnfrdosmothftetwhseatareenqduuaarstdiinocgnusrsvbiexelwocwaos.ngceennetrraattieodns(usoifngst1a/nxdfairtdws.eighTthede Etpnhrgqeo/umgasrtLtaiamonfno.dra1TrsdechraeulcncmuurElvaaqentuedad(utliitronhinneeeaa2.rmcroeaglucrnuetslaostifeodanntahpleayrtaaemmfoeoutuennrdst)o(ifgnaennngael/rmyattLeed,fobbuaysnedtdhineonnAgpn/egaalkfyosartrelsiaov)feturwsaiannrgde Equation 1: Where: AF = Analyte found (ng/mL) = Aliquot Factor /Peaksalorepae - intercept! x AF Equation 2: Analyte found (ppb, ng/g for liver and ng/mL for serum and urine) = Where: DFVF == DFiinluatlioVnoFluamscaatenomarlp,lfeovuonld. (/mngL/m) oLr-)sxamFpVle/mwLei!gxhtD(gF) EFoqruastaiomnp3lecsalfcourltaiftieeddthweiptherckennotwrencoavmeroyu.nts of PFOA and PFOS prior to extraction, Equation 3: Recovery (%) = ((anal, found a(nmoobu)n-taavdgd.eadn(apl,pibn) Ctrl (ppb)) *100% An example of a calculation using an actual sample follows (for PFOS only): Exygen Research Page 17 of 217 : 534 : MIN 313/023622 Exygen Study No.: 023-076 Rat liver sample Exygen ID 0201684 Spk A (Set: 0100802B), fortified at 10 ng/g where: pinetaekrcaerpeta == 53652.2074 slope = 1018.07 dpfaivipnlgbua.tlaiaovdmondltefudaimnc(tefcooorrnttlreovlsel) asalimqupolet fwaceitgohrt ==== == 110 . . . 0 (Not Detected) 210mL 1.0 g From equation 1: Analyte found (ng/mL) = ft 210-1385.0.270741x10 From equation 2: Analyte found (ppb) 5.17 ng/mL <~517 np/ml.x 2 mL x 11 (1.0 g) From e%(q(u1R0a.te3icoopnvp3ebr:-y 0=ppb~) = 10.3 ppb x 100% 10 ppb = 103% 6.6.2 POSF Quantitation and Example Calculation One milliliter of headspace of the sample or calibration standard was injected into the GwstaCeni/gdMhatSred.ds.TlihTneheeaprecaorkengcarereensatsriwaotnaios)nmbwyeaaHssudPreetdCerhamenmdinstehtdaeffisrootnamnsdtoahfretdwecqaurureavteuioswninasgsbgeflieovnweer.caotendce(unstriantgio1n6sc foift Etslhiovqefuetwrastataianornedndppa1rgrodc/gmarlccaLuumrflvoa. etreTsd(ehlrtieuhnnmee' aEaramqnurodeaugtuinrroetinsnosei2f.oancnaalplcayurtlaeamtfeoedutentrhdse)(iangmepnogeu/rnmattLeod,f babanysaeldtyhtoeenfHopuPenadkCaihnreempag)st/uagstiifonognr Equation 1: Analyte found (pg/mL) = (Peak area -1016106011 x DF Where: DF = Dilution Factor slope Exygen Research Page 18 of 217 : 535 : MIN 313/023622 Exygen Study No.: 023-076 Equation 2: Analyte found (ppm, pg/g for liver and pg/mL for serum and urine) - anal, found (ppm! x Standard volume added ('ml.') SV (mL) or SW (g) x VI (mL) . Where: SW = Sample Weight SV = Sample Volume FEcaoqlrucaustliaaomtnepd3lt:ehse fpoerrtcifeinetdrewcoitvheryk.nown amounts of POSF prior to analysis, Equation 3 Recovery (%) = ((anal, found fapmpmou'ln-taavdgd.eadn(apl,pimn)Ctrl (ppm)! xl00% An example of a calculation using an actual sample follows: RwaitthsPeOruSpisdpSSmnleFipatotlaamemuspwnkratecpdihmaoaelaedrenpprerddtelvafee:aovdclEout(loxfmuroymergtee(lnmeavdLIeDd)le)d02(m02L9)87 Volume injected (mL) Spk == ==== == A (Set: 100702A), 4-550011394...011048096050402 fortified at 5 pg/mL From eAqunaaltyiotenf1o:und (pg/mL) = 1494642- -319001 X 1 5480 FArnoamlyteeqfuoautnidon(p2p:m) = = 96.1 pg/mL 96.1 ('ug/mT.l x OOPS ImT.l 0.1 (mL) x 1.0 (mL) From equation 3: = 4.8 pg/mL %Recovery =14.8 nnm 1 x 100% 5 ppm = 96% Exygen Research Page 19 of 217 : 536 : MIN 313/023622 Exygen Study No.: 023-076 Note: This example calculation was done using rounded numbers, and therefore may be slightly different from the values shown in the RAW DATA. 7.0 EXPERIMENTALDESIGN Ebklnaaocnhwk,nstecwtoonocmfesanattrmriaxptilboelnsana(knEsdvfe-orr2oti0rfisesademrauptmlke)sn.ofowErnaLcchCo/nsMacmeSnp/tMlreaStwioaansnsa,elxoytnsrieascftcieeodlndussissaitnmegdptlhoeeffoaorpntpiefriomepdaritaartitxae method and then analyzed in dupHcate. Each set of samples (Ever or serum) for GC/MS analysis consisted of one matrix blank, two matrix blanks fortified at known concentrations, one field sample fortified at a known concentration and ~ 10 samples. 8.0 RESULTS TTmha.belPeTFshOIeVSP-fVFoOuI.nAdTifhnoeuthnPedOciSonFnttrhfooelucnraodtniEtnrvotelhrr,eastceorEunvmtreor,,lasnreadrtuuEmrvi,neera,nssademruuprmlien,seaasnraedmEupsrltieensde aisnraemTEapsbletlesedsairIneEcosrteredspionndTinabglteosthVeTaTnIaXly.te rPeteeanktsionwteirmeesn.ot detected in any of the control samples Individual recoveries for PFOS in the rat Ever, serum and urine samples are detailed in TEvaebrl,esseXru-XmE, LandThuerinaevesraagmeppleesrcwenetrere9c6o%veries11%st,an1d0a2r%d dev1ia4t%io,nsanfdor8P7F%OS i1n1%rat, raersepdeecttaivileeldy.inITndabivliedsuXalIIrIe-cXovVe.riTeshefoarvPerFaOgeApienrctehnet rreactoEvveerri,esserusmta,ndaanrdd udreivnieatsiaomnspfleosr Ps1aF7m%OpA,lersiensaprreaecttdEivevetealryil,.esdeIrinnudmTivaiadbnuldeaslurXrienVceoTvs-aeXmriVepsIlIeIfs.owrTePhrOee S9a9Fv%erinagte9h%pee,rrac1te0n5Et%vreerc,o1sv4eer%ruie,msa,ndans1tda1n0ud%rainrde danevdi9at9i%onsf1o8r%PO, rSesFpienctriavteElyv.er, serum, and urine samples were 99% 18%. 92% 18%, PInFdOivSidiunatlhreesrautltEsvaerer sEasmtepdleisnrTanagbeledXfrIoXm. nPoFnO-dAeteincttehdelreavteElsvteor s5a3m5,p0l0e0s rnagn/gge. d from nnXooXnPI-O.deSPteFFeOdteeSdteilncetvtehedelsirntaoat ns8ey2r7ou0fmtnhgsea/gmr.aptIlEnesdviervarinsdgaumeadlpfrlreeossu.mlItnsnodanirve-iddEeusteatelcdtreeidnsulTeltvaseballsreetXoEXs1t5.e8dT,0ihn0e0Treanbgw/lamesL. IwTnnngoaad/nbsmi-vlndeLioed.XtuPeaIXcOnltIderSVeidFvs.liueddlveuPtsetaFellaOscrrteteSeosdluii1nsiln2tttse,ia1drane0riy0neraonETtgfsua/ttrmbehidnleLeeir.nXastIaTXnsmaedIprbIiuv.lleimePdsXuFrsaaaOXlnmAVrgepe.slidenuPslftt.FrhsoIOenmaAdrreainvtiEonisdnsettu-rheduaedelmrtirenaesctsaTtuuemalrdtibpsnllleeaeevrssXeearlmaEXsnsptHgotleeeId4d.s7ifTn1r0hoemre ranged from non-detected levels to 18,600 ng/mL. Individual results are Ested in Table Exygen Research Page 20 of217 : 537 : MIN 313/023622 Exygen Study No.: 023-076 XXVI. There was no POSF detected in any of the rat urine samples. Individual results are listed in Table XXVII. 9.0 CONCLUSIONS - The rat liver, serum, and urine samples were successfully extracted and analyzed for PFOS, PFOA and POSF according to the appropriate analytical method. 10.0 RETENTION OF DATAAND SAMPLES WREosurxeichsagheecinnataractsclhohfeipanwifcseiiitsnllrliuaotlbymfe-aaesnlpsnlahetlricyaplitowfpiigcecasdd,lraatrthoeawo,ptwoahdsreeatvwtsiaepse,rolcwlneoxasimlosalrpca.btleescTtiegorhe,npitaseiaeidildsncooeooerdfipsgyitninenoomattfhlpitepnheacreEapluxfteuidyrnregaedelfanalatocanRgiaglseiletysywnet-ieriscarlpalaclethbecreideafpirbscocuyrhrbtaiEmvawxeniystddtgeafeadotlna.rl tihne p1e9r9i7o)d, oEfNtiVm/eMsCpe/CciHfiEedMi(n98O)E17C.D PRrientcaiipnleeds osfaGmopoledsLoabforreaftoerryenPcreacstiucbest(aanscreesvisaerde archived by the sponsor. Exygen Research : 538 : Page 21 of217 MIN 313/023622 Exygen Study No.: 023-076 TABLES Exygen Research : 539 Page 22 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table L Summary ofPFOS in Control Rat Liver Samples Sponsor ID Lot#14124 LLoott##1144112244 Lot#14124* Lot#14124 LLoott##1144112244* Lot#14124 LLoott##1144112244* Exygen ID 00220011668844 CCoonnttrrooll AA 0201684 Control A 0201684 Control A 0201684 Control A 00220011668844 CCoonnttrrooll AA 0201684 Control A 00220011668844 CCoonnttrrooll AA Set Number 100802B 110001840022BB 110011640022BA ' 11001166022BA 101602B 1100221I0022AA PFOS Found (ng/g) - ND ND ND NNDD NNDD ND NNDD Table n . Summary of PFOS in Control Rat Serum Samples Sponsor ID LLoott##0077002244* Lot #07024 LLoott##0077002244* Lot#07024* LLoott##0077002244* Lot #07024 Lot#07024* Exygen ID 00220011668822 CCoonnttrrooll AA 0201682 Control A 00220011668822 CCoonnttrrooll AA 000222000111666888222 CCCooonnntttrrrooolll AAA 00220011668822 CCoonnttrrooll AA Set Number 00992200Q022BB 092502A 009922550022AB 011900200511000222BAA 100202A 100202A FounPdF(OngS/mL) NNDD ND NNDD NNNDDD NNDD Table III. Sum m ary of PFOS in Control Rat Urine Samples Sponsor Exygen Set PFOS ID ED Number Found (ng/mL) LLLooottt###000777111222444* 0201683 Control A 00220011668833 CCoonnttrrooll AA 111000000373000222AAA ND NNDD LLoott##0077112244* 00220011668833 CCoonnttrrooll AA 110000770022BA NNDD N*(0DD.0u0=p0LLLlN1iooocotftta##tt#tg00e0D7/77m1I1e1n22tL24je44e)**c)ctteidon(Ar000e222a000111le666s888s333 Control A CCoonnttrrooll AA 100702B 110022110022BB than the lowest concentration of the ND NNDD calibration standards Exygen Research Page 23 of 217 : 540 : MEN 313/023622 Exygen Study No.: 023-076 Table IV. Summary ofPFOA in Control Rat Liver Samples Sponsor ID Lot #14124 Lot#14124* LLLooottt###111444111222444* LLoott##1144112244* Lot#14124* LLoott##1144112244* Exygen ID 0201684 Control A 00220011668844 CCoonnttrrooll AA 00220011668844 CCoonnttrrooll AA 0201684 Control A 0201684 Control A 0201684Control A 00220011668844 CCoonnttrrooll AA Set Number 110000880022BB 101402A 110011460022AA 10162A 110011660022BB 110022110022AA PFOA Found (ng/z) . . . NNDD ND NNDD ND NNDD NNDD Table V. Summary of PFOA in Control Rat Serum Samples Sponsor ID LLLooottt###000777000222444* LLoott##0077002244* LLLooottt###000777000222444** Lot#07024 Lot#07024* Exygen ID 000222000111666888222 CCCooonnntttrrrooolll AAA 00220011668822 CCoonnttrroollAA 000222000111666888222 CCCooonnntttrrrooolll AAA 00220011668822 CCoonnttrroollAA Set Number 000999222050000222BBA 009922550022BA 011900200511000222BAA 110000220022AA PFOA Found (ng/mL) NNDD ND NNDD ND NNDD NNDD Table VI. Summary of PFOA in Control Rat Urine Samples Sponsor Exygen Set PFOA ID ID Number Found (ng/mL) LLLooottt###000777111222444* 0201683 Control A 00220011668833 CCoonnttrroollAA 110000330022AA 100702A ND NNDD LLoott##0077112244* Lot#07124* 0201683 Control A 00220011668833 CCoonnttrrooll AA 100702A 110000770022BB ND NNDD N*(0DD.0u0=p0LLlN1iocooptat#t#gt00eD/77m1Ie1n2tL2je44e)*c)ctteidon(Ar00e22a0011le6688ss33 CCoonnttrroollAA 110022110022BB than the lowest concentration of the NNDD calibration standards Exygen Research Page 24 of 217 : 541 : MIN 313/023622 Exygen Study No.: 023-076 Table VII. Summary of POSF in Rat Liver Control Samples Sponsor ID LLoott##2211882244 LLoott##2211882244 Lot #21824 LLLooottt###222111888222444 Lot #21824 Lot#21824 Exygen ID 00220022887777 000222000222888777777 000222000222888777777 0202877 0202877 Set Number 11Q022230022AA 111000222344000222AAA 111000222855000222AAA 102802A 102902A FouPnOd S(uFg/g) ' NNDD- NNNDDD ND NNDD ND ND Table VH[. Summary of POSF in Rat Serum Control Samples Sponsor ID LLLooottt###000777000222444 LLLooottt###111666444222333 LLLooottt###111666444222333 Lot #07024 Exygen ID 0201682 00220011668822 000222000222999888777 000222000222999888777 0201682 Set Number 092602B 019020710022AA 111000000324000222AAA 110000470022AA 100802A 101702A POSF Found (ug/mL) NNNDDD NNNDDD NNNDDD ND Table IX. Summary ofPOSF in Rat Urine Control Samples Sponsor ID Exygen ID Set Number POSF Found Oig/mL'l LLoott##2200332244 LLLooottt###222000333222444 LLoott##0270132244 Lot #07124 00000222220000022222777778888855555 00220021768853 0201683 110011100022AA 111000111465000222AAA 110022120022AA 102902A NNDD NNNNNDDDDD ND ND = Not Detected Exygen Research Page 25 of 217 : 542 : MIN 313/023622 Exygen Study No.: 023-076 Table X. Summary ofPFOS Fortification Recoveries in Rat Liver Sponsor Exygen Set Amt % ID ID Number Added (ng/g) Recovery LLoott##1144112244 Lot#14124 Lot #14124 21 0201684 Spk A 0201684 SpkA* 00220011668844SSppkkBB 0203485 Spk C 100802B 100802B 110000880022BB 100802B 10 10 * -19023 5500 5000 92 18082 LLoott##211144112244 LLoott##11144112244 Lot #114124 LLLooottt###111444111222444 42 Lot#4124124 LLoott ##1144112244 Lot#14124 6611 LLLooottt###111444111222444 Lot #881114124 000222000311466888544SSSpppkkkACA* 00220011668844SSppkkBB* 00220033446655SSppkkCC* 0201684 Spk A 0201684 Spk A 00220011668844SSppkkBB* 00220033550055SSppkkCC* 0201684 Spk A 00220011668844SSppkkAB 0201684 SpkB* 00220033552244SSppkkCC* 000222000111666888444SSSpppkkkAAB* 0201684 Spk B* 00220033554433SSppkkCC* 100802B 110011440022BB 110011440022BB 11Q011440022BB 101602A 101602A 110011660022AA 110011660022AA 101602B 110011660022BB 101602B 110011660022BB 111000222111000222AAA 102102A 110022110022AA 5000 1100 5500 55000000 10 10 5500 5500000000 10 5100 550050000000 1100 50 50500000 500000 94 18051 110055 112 104 115 19050 1199004214 18083 9682 75 81 110019 988738 STANDARD DAEVVIEARTAIOGNE:: 1916 * Duplicate Injection RELATIVESTANDARD DEVIATION: 11 Exygen Research : 543 : Page 26 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table XI. Summary of PFOS Fortification Recoveries in Rat Serum Sponsor Exygen Set Amt % ID ID Number Added (ng/mL) Recovery Lot#07024 0201682 Spk A 092002B 10' ' ' 135 Lot#07024 0201682 Spk A* LLoott##0077002244 00220011668822SSppkkBB* 1M-GROUP I Day92 0203367 SpkC 092002B 092002B 009922000022BB 1M-GLRoOt#U0P7012D4ay 92 Lot#07024 0203367 SpkC* 00220011668822SSppkkAA* 092002B 009922550022AA 10 5500 5000 50110000 123 126 125 99 99 9714 LLoott##0077002244 00220011668822SSppkkBB* 092502A 092502A 221IFF--GGRROQUUPP 1I DDaayy9922 Lot#07024 0203387 Spk C 00220031368872SSppkkCA* 009922S50022AA 092502B LLoott##0077002244 Lot#07024 0201682 Spk A* 00220011668822SSppkkBB 009922550022BB 092502B 50 50 55000000 10 10 5500 97 111007 9799 98 99 105 4411MM--GGRROOUUPP 1I DDaayy Lot#07024 112211 000222000331446008662SSSpppkkkCAC* 092502B 019020510022BA LLoott##0077002244 00220011668822SSppkkAB* 110000110022AA 5000 501000 5100 111010127 19084 61F-GLRoOtU#0P7102D4ay 121 61F-GROUP 1Day 121 000222000133644822266SSSpppkkkCBC** 110000110Q22AAR 100102AR LLLooottt###000777000222444 000222000111666888222SSSpppkkkABA 111000000222000222AAA 5550000000 10 5100 101 9985 110117 105 Lot#07024 0201682 SpkB* 100202A 8811MM--GGRROOUUPP 11WWeeeekk44 00220033444455SSppkkCC* 110000220022AA 22000050000000 17100 85 AVERAGE: 102 STANDARD DEVIATION: 14 * Duplicate Injection RELATIVE STANDARDDEVIATION: 14 Exygen Research : 544 : Page 27 of217 MIN 313/023622 Exygen Study No.: 023-076 Table XII. Summary ofPFOS Fortification Recoveries in Rat Urine Sponsor ID LLoott##0077112244 LLoott ##0077112244 1 1 LLoott##0077112244 LLoott##0077112244 2244 Lot#07124 LLLooottt###000777111222444 44 44 Lot #07124 LLoott##0077112244 Lot#07124 6644 ExIyDgen Set Amt % Number Added (ne/mL) Recovery 00220011668833SSppkkAA* 110000330022AA 0201683 SpieB 0201683 SpkB* 00220033556633SSppkkCC* 111000000333000222AAAR 100302AR 00220011668833SSppkkAA* 0201683 Spk B 0201683 SpkB* 100702A 100702A 110000770022AA 10 10 5500 .500 500 1100 5500 87 84 84 88 110 18085 83 7849 00220033558866SSppkkCC* 110000770022AA 00220011668833SSppkkAA* 0201683 Spk B 100702B 110000770022BB 00220013668036SSppkiBC* 00220031660863SSppkkCA* 100702B 110000770022BB 102102B 00220011668833SSppkkAB* 110022110022BB 0201683 SpkB* 102102B 00220033662255SSppkkCC* 110022110022BB 550000 10 10 5500 500 500 511000 1150000000 998758 988982 9999 71 7828 67 7786 RELATIVE SSTTAANNDDAARRDDDDAEEVVVIIEAARTTAIIOOGNNE::: 118317 * Duplicate Injection Exygen Research : 545 : Page 28 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table XHI. Summary ofPFOA Fortification Recoveries in Rat Liver Sponsor ID Exygen ID Set Amt % Number Added (ng/g) Recovery LLoott##1144112244 LLoott##1144112244 21 Lot #2114124 Lot#14124 LLoott##1144112244 1 Lot #114124 Lot#14124 LLoott##1144112244 4422 Lot#14124 LLLooottt###111444111222444 6611 LLoott##1144112244 Lot#14124 Lot#S1I4124 SI 00220011668844SSppkkAA* 000222000131646888454SSSpppkkkBBC* 0203485 Spk C* 00220011668844SSppkkAA* 0201684 SpkB 0201684 SpkB* 0203465 SpkC 00220031466854SSppkkCA* 000222000111666888444SSSpppkkkBAB** 000222000313565080545SSSpppkkkCCA* 00220011668844SSppkkAB* 00220013658244SSppkkBC* 0203524 SpkC* 0201684 Spk A 00220011668844SSppkkAB* 0201684 Spk B* 0203543 SpkC 0203543 Spk C* 100802B 100802B 110000880022BB 1100O088O022BB 110011440022AA 111000111444000222AAA 110011460022AA 111000111666000222AAA 101602A 110011660022BA 110011660022BB 101602B 110011660022BB 111000222111000222AAA 102102A 102102A 1Q2102A 1100 50 505000 5000 1100 5500 55000000 10 551000 50000 5010000 10 50 5050000 50000 511000 50 520000 520000 -110092 9869 101 110114 99 104 94 96 19083 111 19070 9972 101 19034 988232 110216 106 18048 83 STANDARD DEAVVIEARTAIOGNE: 99 ' Duplicate Injection RELATIVE STANDARD DEVIATION Exygen Research : 546 : Page 29 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table XIV. Summary ofPFOA Fortification Recoveries in Rat Serum Sponsor Exygen Set Amt % m ID Number Added (ng/mL) Recoven Lot#07024 Lot#07024 00220011668822SSppkkAA* 092002B 092002B 10 ' 10 - - 111115 LLoott##0077002244 . 1M-GROUP l Day 92 0201682 SpkB 00220013638627SSppkkBC 009922000022BBRR 09200?B S5O0 5000 114420 120 1M-GLLRooOtt##U00P7700122D44ay 92 LLoott##0077002244 0203367 SpkC* 0201682 SpkA 000222000111666888222SSSpppkkkABB** 009922050022BA 092502A 009922550022AA 2211FF--GGRROOUUPP 11DDaayy 9922 Lot #07024 000222000331336888772SSSpppkkkCCA* 092502A 009922550022BA LLoott##0077002244 00220011668822SSppkkAB* 009922550022BB Lot#07024 0201682 Spk B* 092502B 4411MM--GGRROOUUPP 11DDaayy Lot #07024 LLoott##0077002244 6611FF--GGLRRoOOtUU#0PP71102DD4aayy 112211 112211 00220033440066SSppkkCC* 000222000111666888222SSSpppkkkABA* 000222000133644822266SSSpppkkkBCC* LLLooottt###000777000222444 Lot#07024 00220011668822SSppkkAA* 0201682 SpkB 0201682 SpkB* 009922550022BB 111000000111000222AAA 100102A 11000011Q022AARR 110000220022AA 100202A 100202A 501000 10 50 505000 501000 10 5500 55000000 10 5100 550000 5000 10 5100 50 18290 9946 100 110010 9818 107 113 109 110 111000334 1780915 102 102 110086 AVERAGE: 105 * Duplicate Injection RELATIVESSTTAANNDDAARRDDDDEEVVIIAATTIIOONN:: 1144 Exygen Research : 547 : Page 30 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table XV. Summary ofPFOA Fortification Recoveries in Rat Urine Sponsor Exygen Set Amt % ID ID Number Added (ug/mL) Recovery LLoott ##0077112244 L o t#07124 Lot#017124 00220011668833SSppkieAA* 00220011668833SSppkkBB* 0203563 Spk C 1I0000330022AA 110000330022AA 100302A 1100 -5550000 T9062 11832003 1 Lot #07124 00220031566833SSppkkCA* 110000370022AA 51000 18025 LLLooottt###000777111222444 24 Lot#2047124 00220011668833SSppkkAB* 0201683 Spk B* 000222000313565888636SSSpppkkkCCA* 111000000777000222AAA 111000000777000222BAA Lot#07124 LLoott##0077112244 4444 0201683 Spk A* 0201683 SpkB 0201683 SpkB* 00220033660066SSppkkCC* 110000770022BB 100702B 110000770022BB LLLooottt###000777111222444 Lot #660447124 0201683 Spk A 00220011668833SSppkkAB* 0201683 Spk B 00220033662255SSppkkCC* 111000222111000222BBB 110022110022BB 102102B 10 5500 550000 10 551000 500 51000 5100 50 11000000 111021338 110032 103 106 112296 106 108 111333300 130 8896 * Duplicate Injection RELATIVE SSTTAANNDDAARRDD DDAEEVVVIIEAARTTAIIOOGNNE::; 1117510 Exygen Research : 548 : Page 31 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table XVI. Summary of POSF Fortification Recoveries in Rat Liver Sponsor ID Lot#1201824 Lot#112101824 Lot #1211824 21 Lot#232121824 Lot#3221824 Lot#442221824 Lot#552221824 Lot #662221824 7733 Lot #8231824 83 Lot #992331824 Exygen ID NuSmetber Amt Added (pg/g) % Recovery 00220023847774SSppikeAB 110022220022AA 55 -110066 000222000323844777754 SSSpppfkkcBCA 000222000233844778755 SSSpppkkkCBA 111000222233000222AAA 102302A 110022330022AA 55 _ 5555 111113573 111222600 000222000323484879575 SSSpppkkkACB 111000222344000222AAA 555 969329 00220032489757 SSppkk AC 11002244O022AA 55 19011 000222000233855700755SSSpppkkk ABC 111000222445Q00222AAA 555 1983300 0203515 Spk B 00000222220000032323585851727257575 SSSSSpppppkkkkkABCAC 111000222555000222AAA 111Q00222585000222AAA 555 555 11911589 888357 00220033553355 SSppkkBC 110022880022AA 55 110020 00220023857475 SSppkk AB 110022880022AA 55 8845 0203545 Spk C 102802A 00220023857575 SSppkkAB 0203555 Spk C 111000222999000222AAA 55 55 8772 7796 STANDARD DAEVVIEARTAIOGNE RELATIVE STANDARD DEVIATION 911989 Exygen Research : 549 : Page 32 of217 MIN 313/023622 Exygen Stndy No.: 023-076 Table XVII. Summary ofPOSF Fortification Recoveries in Rat Serum Sponsor Exygen Set Amt % ID ID Number Added (pg/mL) Recovery 16M-GLRotOU07P0424Day 92 J1267MF--GGLRRoOOt#UU0P7P0242D4Daayy9922 00000222220000011333663338889822222SSSSSpppppkkkkkBBAAC 009922660022BB 000999222767000222BAA . 555S5 111135 1781760 27F-GLRoOt#U0P7022D4ay 92 88MM--GGRROOUUPP22 DDaayy9922 3322FF--GGLRRoOOt#UU1PP64332DD3 aayy 9922 45M-GLRoOt#U1P6412D3ay 121 45M-GLRoOt#U1P6412D3ay 121 0203392 SpkC 00220013638724 SSppkk BA 000222000323933879747 SSSpppkkkABC 00220032938977 SSppkkCA 000222000233944811700 SSSpppkkk BAC 019020710022AA 111000000112000222AAA 100202A 110000320022AA 111000000343000222AAA 5511MM--GGrroouupp33DDaayy 112211 66771122FFFF----GGGGLLrrrroooooottuuuu##pppp1166331144DDDD22aaaa33yyyy 112211 112211 000000222222000000232333949444818221767666 SSSSSSppppppkkkkkk BBACCA 00220033443366 SSppkkBC 111111000000000000444447000000222222AAAAAA 110000770022AA 9977FF--GGLorrootuu#pp16444WW23eeeekk44 8844MM--GGLRRoOOt#UU0PP70112WW4 eeeekk44 0202987 SpkA 00000222220000033133446446684411288 SSSSSpppppkkkkkBBACC 100802A 111110000000111887770000022222AAAAA 555 S5 555 555 555555 5 55 55 555 1971721 888840 9800 5668 187813642 1893569 100 110020 19976158641 RELATIVESSTTAANNDDAARRDD DDAEEVVVIIEAARTTAIIOOGNNE::: 129802 Exygen Research : 550 : Page 33 of217 MIN 313/023622 Exygen Study No.: 023-076 Table XVHL Summary ofPOSF Fortification Recoveries in Rat Urine SpoIDnsor Lot#210324 Lot#210324 Lot#21121140324 Lot#242340324 43 Lot #20324 Lot#662440324 Lot #77305547124 Lot#3047124 5511 Exygen Set Amt % ID Number Added (ug/mL) Recovery 0202875 Spk A 0000222200003233855576765333SSSSppppkkkk BBCA 000222000233855778536 SSSpppkkk CBA 000222000233865708556SSSpppkkk BCA 00220032680755 SSppkkCA 000222000233866722555 SSSpppkkkCBA 000222000133666833366 SSSpppkkkBAC 000222000133655899366 SSSpppkkk BCA 00220033661133 SSppkkBC 110011000022AA 111000111101000222AAA 111000111441000222AAA 111000111455000222AAA 110011560022AA 111000112661000222AAA 111000222121000222AAA 111000222229000222AAA 110022990022AA 55 55 . 5555 555 55 5 55555 5 55 55 -111254 111122533 11721815 11811229 103 77 11900174 9634 71 988338 9910 AVERAGE: 99 RELATIVE SSTTAANNDDAARRDD DDEEVVIIAATTIIOONN:: 18 18 Exygen Research : 551 : Page 34 of217 MIN 313/023622 Exygen Study No.: 023-076 Table XIX. Summary of PFOS Residues in Rat Liver Samples Sponsor ID 1 1* 22* 3. 3* 44* 55* 678678** 9 91*0 10 11 1112* 12* 111334* 14* 15 111566** 111778* 1189* 1290* 20* Exygen ID 00220033446655 0203466 00220033446667 00220033446678 0203468 00220033446699 0203470 00220033447710 000222000333444777122 0203473 00220033447734 0203474 0203475 00220033447756 0203476 000222000333444777778 00220033447789 0203479 00220033448800 0203481 00220033448812 00220033448832 0203483 00220033448844 Set Number 101402B 101402B 101402B 101402B 101^02B . 110011440022BB 101402B 110011440022BB 110011440022BB 101402B 111000111444000222BBB 101402B 110011440022BB 101402B 101402B 110011440022BB 1014023 111110000011111444440000022222BBBBB 101402B 110011440022BB 111000111444000222BBB 110011440022BB 101402B 110011440022BB PFOS Found (ng/g) * 117986 117632 225 234 211968 236792 44700 4469540000 4602300000 58200 333869289000000 37200 110000 105000 110014000000 111212246000000000 127000 91800 86200 335128000000 222513068000000000 229000 229673000000 221192000000 N* DDu=pNlicoatteDIentjeecctteidon(Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 35 of 217 : 552 : MIN 313/023622 Exygen Study No.: 023-076 Table XIX (coni'). Summary of PFOS Residues in Rat Liver Samples Sponsor m 2211 2222* 24 24* 2255 222676 27* 28 2289* 29* 3300* 31 333122** 3333* 3344* 35 333566** 3377* 38 3389* 39* 4400* Exygen m 00220033448855 0203486 00220033448876 00220033448878 0203488 000222000333444889990 00220033449910 00220033449912 0203492 000222000333444999334 00220033449954 0203495 00220033449966 00220033449977 0203498 000222000333444999899 0203500 0203500 000222000333555000112 0203502 00220033550033 NuSmetber 110000880022BB 211000000888000222BBB 100802B ' 110000880022BB 100802BR 110000880022BBRR 110000880022BBRR 111000000888000222BBBRRR 111000000888000222BBBRRR 110000880022BBRR 100802BR 110000880022BBRR 110000880022BBRR 100802BR 110000880022BBRR 100802BR 110000880022BBRR 111000000888000222BBBRRR 100802BR 110000880022BBRR FouPnFdO(nSg/g) ' 228962. .284 227904 296 163 126 9920500000 71700 6748310000 66000 8921860000 60900 56600 227000 211466000000 111486519000000000 210966000000 253000 255470251000000000 222547000000 359000 339430000000 322587667000000000 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 36 of 217 : 553 : MIN 313/023622 Exygen Study No.: 023-076 Table XIX (coni'). Summary ofPFOS Residues in Rat Liver Samples xmsor Exygen Set PFOS ID ID Number Found (na/^l 4411* 444232* 0203504 0203504 00220033550055 0203506 100802BR 100802BR 110011660022AA 101602A ' 1790 . 1231781821330 4434* 44* 444556* 46* 47 0203506 00220033550077 000222000333555000889 0203509 0203510 I01602A " 110011660022AA 111000111666000222AAAR 101602AR 101602AR 116263 123 110063 30000 2380590000 4478* 48* 00220033551110 0203511 101602AR 110011660022AARR 442558913000000 445990* 50* 51 555122** 555334* 555455** 5566* 57 5578* 58* 5599* 6600* 0203512 00220033551123 000222000333555111344 00220033551155 0203516 00220033551176 0203517 00220033551188 0203519 0203519 00220033552200 0203521 00000222220000033333555552222213322 110011660022AARR 101602AR 110011660022AARR 111000111666000222AAARRR 101602AR 110011660022AARR 111000111666000222AARRR 101602AR 101602AR 111000111666000222AAARRR 111110000011111666660000022222AAAAARRRRR 442856377000000 2972760000 9851550000 75900 6517440000 83200 19007400000 97600 223240000000 111429046000000000 200000 225376000000 114598000000 N*DD=upNliocat tDeeItnejcetcetdio(nArea less than lowest calibration standard of 0.0001 |ig/mL) Exygen Research Page 37 of217 : 554 : MIN 313/023622 Exygen Study No.: 023-076 Table XIX (cont'). Summary ofPFOS Residues in Rat Liver Samples Sponsor ID 61 61* 62 6624* 6645* 666566** 666778* 68* 69 69* 7700* 71 7712* 777233** 7744 777556* 7767* 77* 7788* 79 79* 80 80* Exygen ID 0203524 0203524 0203525 00220033552256 00220033552276 0203527 00220033552288 0203529 00220033552390 0203530 0203531 000222000333555333122 000222000333555333334 00220033553354 0203535 Q0220033553366 0203537 00220033553378 0203538 00220033553399 000222000333555444100 0203541 0203542 0203542 Set Nnmber 110011660022BB 111000111666000222BBB . 110011660022BB 101602B 110011660022BBRR 111000111666000222BBBRRR 101602BR 101602BR 111000111666C00222BBBRRR 111000111666000222BBBRRR 101602BR 110011660022BBRR 101602BR 110011660022B3RR 110011660022BBRR 111000111666000222BBBRRR 111000111666000222BBBRRR 101602BR 101602BR 101602BR PFOS Found (ng/g) 115459 111225129 156 136 111 4548620000 2268040000 33100 4312190000 4302300000 29300 93700 110510000000 110481000000 116000 111264000000 7899790000 158000 111768269000000000 221256423000000000 214837000000 248000 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 pg/m L) Exygen Research Page 38 of217 : 555 : MIN 313/023622 Exygen Study No.: 023-076 Table XIX (cont'). Summary of PFOS Residues in Rat Liver Samples Sponsor ID 8811* 82 8823* 8834* 84* 8855* 86 8867* 87* 8888* 898909* 9901* 91* 9922* 999334* 9945* 95* 9966* 999778* 9989* 1910900*0* Exygen ID 00220033554433 00220033554444 0203545 00220033554456 00220033554476 0203547 00220033554488 0203549 000222000333555455900 0203551 00220033555512 00220033555532 0203553 00220033555544 0203555 0203555 0203556 00220033555576 00220033555578 0203558 000222000333555556990 00220033556610 000222000333555666122 Set Number 110022110022AA 102102A 102102A 102102A. 1I0022110022AA 111000222111000222AAA 111000222111000222AAARRR 111000222111Q00222AAARRR 102102AR 102102AR 111000222111000222AAARR 102102A 102102A 102102A 110022110022AA 1Q2102A 102102A 102102A 110022110022AAR 102102AR 110022110022AARR 102102AR 110022110022AARR 11100022211I000222AAARRR PFOS Found (ng/g) 5610..39 5598..30 137 125 50.3 788746...460 173000 211749000000 219000 115539000000 113344000000 117755990090000 11610313200 4547..42 75.3 7223..72 17.6 210794000000 150000 210347000000 212882000000 111943305000000000 N* DDu=plNicoatteDIentejecctteidon(Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 39 of217 : 556 : MIN 313/023622 Exygen Study No.: 023-076 Table XX. Summary ofPFOA Residues in Rat Liver Samples Sponsor ID 1 1* 2 23* 3* 44* 5 5* 676* 788** 9 9* 1100 11 111212** 111334* 14* 15 15* 1166* 111778* 1189* 212090** Exygen ID 0203465 0203465 00220033446666 0203467 0203467 0203468 00220033446689 0203469 000222000333444777100 00220033447712 0203472 00220033447733 000222000333444777544 000222000333444777566 000222000333444777778 00220033447798 000222000333444788900 000222000333444888112 00220033448832 000222000333444888344 Set Number 101402A 101402A 101402A 110011440022AA. 110011440022AA 1Q14Q2A 110011440022AA 10I402A 110011440022AA 101402A 110011440022AA 110011440022AA 101402A 110011440022AA 101402A 110011440022AA 111000111444000222AAA 101402A 101402A 110011440022AA 101402A 111000111444000222AAA 110011440022AA 111000111444000222AAA PFOA Found (ng/g) 118 118111226856 169 109 110745 172 11333500 1480 111555739000 657 653 1230 21824100 22652700 22987200 2820 33332600 1790 661997677000 5850 55251700 5070 6200 S65323522000 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 [ig/mT .) Exygen Research Page 40 of 217 : 557 : MIN 313/023622 Exygen Study No.: 023-076 Table XX (cont'). Summary of PFOA Residues in Rat Liver Samples jonsor n> Exygen . ID Set Number PFOA Found (ns/z) 2211* 0203485 0203485 110000880022BB 2332..27 2222* 2424* 25 222566** 222778* 222899** 30 30* 0203486 0203486 0203487 0203487 0203488 000222000333444888899 00220033449900 0203491 000222000333444999122 00220033449933 110000880022BB 100802B 100802B 100802B 110000880022BB 100802B 110000880022BB 100802B 111000000888000222BBB 110000880022BB 333112* 000222000333444999454 111000000888000222BBB 333233** 34 3345* 3356* 36* 3377* 000222000333444999566 000222000333444999877 0203498 0203499 0203499 00220033550000 111000000888000222BBB 111000000888000222BBB 100802B 100802B 100802B 110000880022BB 333889* 434090** 00220033550011 0203502 00220033550023 0203503 111110000000000888880000022222BBBBB 100802B 3365..53 38.8 3390..84 222698...115 34.7 4344..39 453111..214 34.9 33.5 433091..215 333479...559 44.8 4319..39 44.9 95.1 103 5586..86 456913...116 5444..90 41.0 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 41 of 217 : 558 : MIN 313/023622 Exygen Study No.: 023-076 Table XX (cont'). Summary of PFOA Residues in Rat Liver Samples OO Sponsor ID 41 41* 4422* 43 , 444344** 4455* 4466* 47 4478* 4499* 5500* 51 S552I2** 555334* 54* 55 5556* 56* 5577* 58 5589* 59* 6600* ExEyDgen 0203504 0203504 0203505 00220033550056 0203506 0203507 00220033550078 0203508 00220033550099 0203510 000222000333555111110 0203512 0203512 000222000333555111334 00220033551154 0203515 000222000333555111766 00220033551178 000222000333555111899 0203520 00220033552210 0203521 0203522 00220033552232 0203523 Set Number 110000880022BB 101602A 110011660022AA. 110011660022AA 1016Q2A 110011660022AA 111000111666000222AAA 101602A 110011660022AA 101602A 101602A 110011660022AA 101602A 101602A 110011660022AA 111000111666000222AAA 110011660022AA 101602A 101602A 101602A 111000111666000222AAA 110011660022AA 111000111666000222AAA PFOA Found (ng/g) 6688..04 7872..59 63.1 7444..64 44.5 5528..13 194 315716 355 229953 417 439 446327302 634858 381 448892 221164 574 564 993380 433094829 340 272 267 335676 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 pgfmL) Exygen Research Page 42 of217 : 559 : MIN 313/023622 Exygen Study No.: 023-076 Table XX (cont'). Summary ofPFOA Residues in Rat Liver Samples Sponsor ID 61 6612 6624* 64* 65 65* 6666* 666778* 6689* 69* 7700* 7711* 72 777233** 74 74* 75 7756* 777677** 777889* 788900** Exygen ID 0203524 0203524 0203525 00220033552256 000222000333555222767 0203528 0203528 00220033552299 0203530 00220033553310 0203531 00220033553322 0203533 00220033553334 000222000333555333455 00220033553366 0203537 00220033553378 0203538 00220033553399 000222000333555444100 00220033554412 0203542 Set Number 110011660022BB 111000111666000222BBB . 110011660022BB 101602B 101602B 10I602B 110011660022BB 101602B 110011660022BB 101602B 101602B 110011660022BB 110011660022BB 111000111666000222BBB 101602B 110011660022BB 110011660022BB 101602B 110011660022BB 111000111666000222BBB 110011660022BB 101602B PFOA Found (ne/g) ' 3346.30 222302...576 ND 2272..50 222600...694 2189..77 2291..79 24.0 3343..74 2108..88 ND 223574...173 232752...136 19.4 2179..06 21.2 2N1D.5 3N1D.6 30.0 2289..76 * Duplicate Injection ND - Not Detected (Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 43 of 217 : 560 : MIN 313/023622 Exygen Study No.: 023-076 Table XX (cont'). Summary ofPFOA Residues in Rat Liver Samples Sponsor ID 8811 82 8823 83 8844* 8855 888667* 888788** 8899* 90 9901* 999122** 93 9934* 9945* 95* 9999976678** 9989* 99* 110000* Exygen ID 0203543 0203543 00220033554444 0203545 0203545 000222000333555444676 0203547 00220033554488 0203549 00220033554590 00220033555510 0203551 0203552 00220033555523 000222000333555555344 00220033555555 0203556 00220033555576 00220033555578 0203558 000222000333555556990 000222000333555666110 00220033556622 Set Number 102102A 102102A 110022110022AA 102J02A. 102102A 110022110022AA 110022110022AA 102102AR 11Q022110022AARR 102102AR 110022110022AARR 110022110022AARR 102102AR 110022110022AAR 111000222111000222AAA 110022110022AA 102102A 110022110022AA 102102A 110022110022AA 110022110022AA 102102A 102102A 102102A 110022110022AA 102102A PFOA Found (ng/g) ' 4442..70 444703...992 4441..35 44.6 6514..70 5750 57935200 6950 55658900 77543500 8250 86207.40 56.7 7891..39 ND NNDD NNDD 22N46D.38 2119.39 25.7 2199.36 22N44D.31 N* DIm=plNicoatteDIentjeecctteidon(Area less than lowest calibration standard of 0.0001 |ig/mL) Exygen Research Page 44 of 217 : 561 : MIN 313/023622 Exygen Study No.: 023-076 Table XXI. Summary ofPOSF Residues in Rat Liver Samples Sponsor ID 11* 22 3 .. 434* 556* 67* 7* 88* 19190*0 111112* 12* 111334* 14* 1155* 111667* 111788** 1199* 2200* Exygen ED 00220033446655 0203466 00220033446676 000000222222000000333333444444666667788990 00220033447710 00220033447712 00220033447732 00220033447734 000222000333444777554 00220033447766 00220033447777 00220033447788 00220033447799 000222000333444888100 00220033448812 000222000333444888332 00220033448844 Set Number 110Q22220022AA 111000222222000222AAA. 111111110000000022222222222222220000000022222222AAAAAAAA 110022220022AA 102202A 111000222222000222AAA 111000222233000222AAA 110022330022AA 110022330022AA 102302A 111000222333000222AAA 111000222333000222AAA 110022330022AA 111000222333000222AAA 110022330022AA POSF Found (nz/e) ' NNDD ND NNDD NNNDDD NNDD NNNDDD NNNDDD NNNDDD NNNDDD NNNNNDDDDD NNNDDD NNDD NNDD NNDD NNNNDDDD N* DDu=plNicoatteDAetleiqctueodt(Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 45 of 217 : 562 : MIN 313/023622 Exygen Study No.: 023-076 Table X X I (cont'). Summary of POSF Residues in Rat Liver Samples Sponsor ID 2211* 222224* 222545** 222676* 2222287989*** 333010* 3312* 333233** 333445* 333566** 333778* 3389* 434090** Exygen ID 00220033448855 00220033448866 0203487 000222000333444888788 00220033448899 0000222200003333444499991100 00220033449922 00000222220000033333444449999934534 00220033449956 000222000333444999776 0203498 000222000333444999899 000222000333555000100 00220033550012 000222000333555000332 Set Number 110022330022AA 110022330022AA 102302A _ 111000222333000222AAA 110022330022AA 110022330022AA 110022330022AA 110022330022AA 111110000022222333430000022222AAAAA 111100002222444400002222AAAA 110022440022AA 111000222444000222AAA 111000222444000222AAA 11002244O022AA 111000222444000222AAA POSF Found (pg/g) ' NNDD NNDD NNDD NNDD NNDD NNNDDD NNNDDD NNNDDD NNNDDD NNDD NNNDDD NNNDDD NNNDDD NNDD NNNDDD * Duplicate Aliquot ND = Not Detected (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 46 of 217 : 563 : MIN 313/023622 Exygen Study No.: 023-076 Table XXI (coni'). Summary ofPOSF Residues in Rat Liver Samples Sponsor IP 4411 444232* 4434 44 4455 444676 444445789890 50 51 555212* 555334 555455** 555676* 5578* 5565689090** Exygen IP 00220033550044 000222000333555000556 0203506 0000222200003333555500007788 00000222220000033333555551001119900 0000222200003333555511111322 00220033551134 00220033551154 0000222200003333555511115766 000222000333555111788 000222000333555112990 00220033552210 00000222220000033333555552222213322 Set Number 110022440022AA 111000222444000222AAA102402A 11Q022440022AA 110022440022AA 111110000022222444440000022222AAAAA 111100002222444400002222AAAA 110022440022AA 111000222545000222AAA 110022550022AA 111100002222555500002222AAAA 111000222555000222AAA 110022550022AA 111110000022222555550000022222AAAAA POSF Found (ng/g) ND NDNNNDDD ND NNDD NNDD NNNDDD NNNDDD NNNNDDDD ND NNNDDD NNNDDD NNNDDD NNDD NNNNNNDDDDDD NNDD * Duplicate Aliquot ND = Not Detected (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 47 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table XXI (cont'). Summary ofPOSF Residues in Rat Liver Samples Sponsor ID 6611 62 666244* 65 666566** 6677* 66669889** 777010* 71* 72 777233** 777454* 7756* 7767* 7778* 7789* 788900** Exygen . ID 00220033552244 000222000333555222556 00220033552267 00000222220000033333555552222289978 00220033553300 00000222220000033333555553333311322 00220033553334 000000222222000000333333555555333333556746 000222000333555333788 00220033553399 000222000333555444100 000222000333555444122 Set Number 102502A 102502A 111000222555000222AAA. 110022550022AA 111000222555000222AAA 110022550022AA 102502A 111000222555000222AAA 111000222555000222AAA 1I0022550022AA 102502A 111110000022222888880000022222AAAAA 111000222888000222AAA 102802A 102802A 111000222888000222AAA 111000222888000222AAA POSF Found fu/g'i ` NNDD ND NNDD NNDD NNNDDD NNNDDD NNNDDD NNNDDD NNDD NNNDDD NNNDDD NNNDDD NNDD NNDD NNNDDD ND * Duplicate Aliquot ND = Not Detected (Area less than lowest calibration standard of 25 fig/mL) Exygen Research Page 48 of 217 : 565 MIN 313/023622 Exygen Study No.: 023-076 Table XXI (cont'). Summary of POSF Residues in Rat Liver Samples Sponsor m 8811* 888223* 83* 84 8845* 88888888576786**** 8899* 999010* 999122** 999343* 999455** 9966* 999778* 999989** 100 100* Exygen - ID 00000222220000033333555554444433544 00220033554456 0203546 00220033554477 000222000333555444889 000222000333555455900 000222000333555555112 00220033555532 000222000333555555344 00220033555555 00220033555566 00220033555577 00220033555588 000222000333555655099 000222000333555666110 0203562 0203562 Set Number 110022880022AA 111000222888000222AAA102802A 1Q2802A 111000222888000222AAA 111111000000222222888888000000222222AAAAAA 110022880022AA 111000222888000222AAA 112100002222888900002222AAAA 110022990022AA 111000222999000222AAA 102902A 102902A 111000222999000222AAA 111000222999000222AAA 102902A 102902A POSF NNDD' NNNDDD NNDD NNNDDD NNNDDD NNNDDD NNDD NNNDDD NNNDDD NNNDDD NNNDDD NNDD NNNNDDDD NNDD NNDD N* DDu=plNicoatteDAetleicqtueodt (Area less than lowest calibration standard of 25 pg/mL) Exygcn Research Page 49 of 217 : 566 : MIN 313/023622 Exygen Study No.: 023-076 Table XXII. Summary ofPFOS Residues in Rat Serum Samples Sponsor ID 11MM--GGRROOUUPP11DDaayy9922* -223MMMG--GGRRROOOUUUPPP111DDDaaayyy999222 3M-GROUP 1Day 92* 4M-GROUP 1Day92 45MM--GGRROOUUPP11DDaayy9922* SM-GROUP 1Day 92* 667MMM---GGGRRROOOUUUPPP222DDDaaayyy999222* 78MM--GGRROOUUPP22DDaayy9922* 8M-GROUP2 Day92* 9M-GROUP2 Day 92 9M-GROUP2 Day92* 1100MM--GGRROOUUPP22DDaayy9922* 11112121MMMM----GGGGRRRROOOOUUUUPPPP3333DDDDaaaayyyy99992222** 1133MM--GGRROOUUPP33DDaayy9922* 14M-GROUP3 Day 92 1145MM--GGRROOUUPP33DDaayy9922* 1156MM--GGRROOUUPP34DDaayy9922* 16M-GROUP4 Day 92* 111778MMM---GGGRRROOOUUUPPP444DDDaaayyy999222* 111989MMM---GGGRRROOOUUUPPP444DDDaaayyy999222* 2200MM--GGRROOUUPP44DDaayy9922* Exygen ID 00220033336677 000222000333333666889 0203369 0203370 0203370 00220033337711 00220033337722 0203373 000222000333333777344 00220033337755 0203376 00220033337776 0203377 00220033337788 0203379 00220033337890 00220033338810 00220033338812 0203382 000222000333333888334 0203384 0203385 000222000333333888566 Set Number 092002B 009922000022BB 009922000022BB _ 009922000022BB 009922000022BB 092002B 000999222000000222BBBRRR 009922000022BBRR 092002BR 092002BR 000099992222000000002222BBBBRRRR 000999222000000222BBBRRR 092002BR 009922000022BBRR 009922000022BBRR 000999222000000222BBBRRR 000999222000000222BBBRRR 092002BR 092002BR 000999222000000222BBBRRR PFOS Found (ng/mL) ND ND NNNDDD ND 5N0D.6 64.7 64.7 66871300 66258200 55265300 88872600 2553852730000 222343788000000 23800 2230440000 20800 19700 12022400000 199908887814000000000 87400 110000 104000 5555790000 N* DDu=pNlicoattDe Ientejecctteidon(Area less than lowest calibration standard of 0.0001 |ig/mL) Exygen Research Page 50 of 217 : 567 : MIN 313/023622 Exygen Study No.: 023-076 Table XX (cont')< Summary ofPFOS Residues in Rat Serum Samples Sponsor Exygen ED _ ID 2211FF--GGRROOUUPP11DDaayy9922* 00220033338877 22F-GROUP1Day 92 -2224FF--GGRROOUUPP11DDaayy9922* 000222000333333888889 24F-GROUP 1Day 92* 2255FF--GGRROOUUPP11DDaayy9922* 000222000333333899900 2266FF--GGRROOUUPP22DDaayy9922* 00220033339911 27F-GROUP2 Day92 0203392 2278FF--GGRROOUUPP22DDaayy9922* 28F-GROUP 2 Day 92* 2299FF--GGRROOUUPP22DDaayy9922* 3300FF--GGRROOUUPP22DDaayy9922* 333112FFF---GGGRRROOOUUUPPP333DDDaaayyy999222* 0203392 0203393 00220033339934 0203394 00220033339955 0203396 00220033339976 333233FFF---GGGRRROOOUUUPPP333DDDaaayyy999222** 0203397 00220033339988 333445FFF---GGGRRROOOUUUPPP333DDDaaayyy999222* 00220033339999 0203400 3356FF--GGRROOUUPP34DDaayy9922* 00220033440010 3367FF--GGRROOUUPP44DDaayy9922* 00220033440012 37F-GROUP4 Day92* 0203402 38F-GROUP 4 Day 92 3389FF--GGRROOUUPP44DDaayy9922* 000222000333444000334 39F-GROUP4 Day92* 44Q0FF--GGRROOUUPP44DDaayy9922* 00220033440054 0203405 Set Number 009922550022AA 092502A 009922550022A. 0000999922225S5500002222AAAAR 092502AR 009922550022AARR 092502AR 092502AR 009922550022AARR 000099992222555500002222AAAARRRR 092502AR 011900200511000222AAAR 092502AR 092502AR 009922550022AARR 092502AR 092502AR 009922550022AARR 092502AR 009922550022AARR 000999222555000222AAARRR FounPdF(OngS/mL) NNDD ND NNDD ND NNDD 98277000 6350 76938200 8750 88448500 8760 8990 4410520000 39381041149300000000 224354931000000 4745010000 8728910000 84100 150000 115181000000 104000 115326000000 N* DDu=plNicoatteDIentjeecctteidon(Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 51 of 217 : 568 : MIN 313/023622 Exygen Study No.: 023076 Table XXII (coni'). Summary ofPFOS Residues in Rat Serum Samples Sponsor Exygen ID . ID 41M-GROUP 1Day 121 0203406 41M-GROUP 1Day 121* 0203406 4422MM--GGRROOUUPP11DDaayy112211* 00220033440077 43M-GROUP 1Day 121 0203408 43M-GROUP 1Day 121* 44444545MMMM----GGGGRRRROOOOUUUUPPPP1111DDDDaaaayyyy111122221111* 00220033440089 0203409 00220033441100 444667MMM---GGGRRROOOUUUPPP222DDDaaayyy111222111* 000222000333444111112 4478MM--GGRROOUUPP22DDaayy112211* 00220033441132 4489MM--GGRROOUUPP22DDaayy112211* 49M-GROUP2 Day 121* 0203413 00220033441144 5500MM--GGRROOUUPP22DDaayy112211* 00220033441155 51M-GROUP 3 Day 121 0203416 555122MMM---GGGRRROOOUUUPPP333DDDaaayyy111222111** 53M-GROUP 3 Day 121 5534MM--GGRROOUUPP33DDaayy112211* S545MM--GGRROOUUPP33DDaayy112211* 555566MMM---GGGRRROOOUUUPPP434DDDaaayyy111222111** 00220033441167 0203417 000222000333444111889 0203419 0203420 000222000333444222110 57M-GROUP4 Day 121 5578MM--GGRROOUUPP44DDaayy112211* 58M-GROUP4 Day 121* 56569090MMMM----GGGGRRRROOOOUUUUPPPP4444DDDDaaaayyyy111122221111** 00220033442222 0203423 00220033442234 000222000333444222554 Set Number 009922550022BB 092502B 092503 092502B 092503 092503 092503 009922550033 009922550033RR . 009922550033RR 092503R 092503R 009922550033RR 009922550033RR 092503R 000999222555000333RRR 092503R 009922550033RR 009922550023BRR 092503R 009922S50033RR 009922550033RR 009922550033RR 092502BR 011900200522000223AAR PFOS Found (ng/mL) NNDD NNDD ND NNDD ND NNDD 676708930000 47243200 4310 44223900 22438700 11600 1108460000 17700 221459933000000 18800 16500 761788370000000 223683877000000 3335180000 344379771000000 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 52 of 217 MIN 313/023622 Exygen Study No.: 023-076 Table XXII (cont'). Summary of PFOS Residues in Rat Serum Samples Sponsor ID 6611FF--GGRROOUUPP11DDaayy112211 62F-GROUP 1Day 121 62F-GROUP 1Day 121* 64F-GROUP 1Day 121 64F-GROUP 1Day 121* 6655FF--GGRROOUUPP11DDaayy112211* 6666FF--GGRROOUUPP22DDaayy112211* 67F-GROUP2 Day 121 666788FFF-G-GGRRROOOUUUPPP222DDDaaayyy111222111** 69F-GROUP2 Day 121 69FGROUP 2 Day 121* 7700FF--GGRROOUUPP22DDaayy112211* 71F-GROUP 3Day 121 7712FF--GGRROOUUPP33DDaayy112211* 72F-GROUP 3 Day 121* 7733FF--GGRROOUUPP33DDaayy112211* 74F-GROUP 3 Day 121 7745FF--GGRROOUUPP33DDaayy112211* 777566FFF---GGGRRROOOUUUPPP434DDDaaayyy111222111** 7777FF--GGRROOUUPP44DDaayy112211* 78F-GROUP4 Day 121 7789FF--GGRROOUUPP44DDaayy112211* 79F-GROUP 4 Day 121* SOI'-GROUP4 Day 121 80F-GROUP4 Dav 121* Exygen ID 000222000333444222676 0203427 0203428 000222000333444222899 0203430 00220033443310 0203431 0000222200003333444433333223 000222000333444333445 000222000333444333566 0203437 0203437 00220033443388 0203439 000222000333444344900 0203441 00220033444412 00220033444432 000222000333444444344 Set Number 100102A 110000110022AA 110000110022AA 100102A' 110000110022AA 110000110022AARR 100102AR 110000110022AARR 111000000111000222AAARRR 111000000111000222AAARRR 111000000111000222AAARRR 100102AR 100102AR 110000110022AARR 100102AR 110000110022AARR 100102AR 100102AR 110000110022AARR 100102AR I001Q2AR 111000000111000222AAARRR PFOS Found (na/mL) ' ' ' 5NN0DD.9. NNDD NNNDDD 1880531610000 180538000 1110260000 10500 3983114200000 232625177000000 34300 34300 1165540000 32900 11333513600000000 8860080000 72900 7783390000 666749372000000 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 53 of 217 : 570 : MIN 313/023622 Exygen Study No.: 023-076 Table XXII (cont'). Summary ofPFOS Residues in Rat Serum Samples Sponsor Exygen ID ID 81M-GROUP 1Week 4 0203445 81M-GROUP 1Week4* 0203445 8822MM--GGRROOUUPP11WWeeeekk44* 00220033444466 -888343MMM---GGGRRROOOUUUPPP111WWWeeeeeekkk444* 84M-GROUP 1Week4* 0203447 0203447 00220033444488 85M-GROUP 1Week4 85M-GROUP1 Week 4* 00220033444499 888667MMM---GGGRRROOOUUUPPP444WWWeeeeeekkk444* 00220033445500 0203451 888788MMM---GGGRRROOOUUUPPP444WWWeeeeeekkk444** 89899G90MMMM----GGGGRRRROOOOUUUUPPPP4444WWWWeeeeeeeekkkk4444** 91F-GROUP 1Week4 91F-GROUP1 Week 4* 9922F1--GGRROOUUPP11WWeeeekk44* 93F-GROUP1 Week 4 9934FF--GGRROOUUPP11WWeeeekk44* 94F-GROUP 1Week4 99S5FF--GGRROOUUPP11WWeeeekk44* 00220033445512 0203452 000222000333444555334 0203454 0203455 0203455 00220033445566 00220033445577 0203458 0203458 00220033445599 9966FF--GGRROOUUPP44WWeeeekk44* 9977F5-:-GGRROOUUPP44WWeeeekk44* 981-GROUP 4 Week4 999989FFF---GGGRRROOOUUUPPP411WWWeeeeeekkk444** 11O0O0FF--GGRROOUUPPI1WWeeeekk44* 0203460 0203460 000222000333444666112 00220033446632 000222000333444666344 Set Number 100202A 100202A 111000000222000222AAA. 110000220022AA 100202A 100202A 100202A 111000000222000222AAARRR 111000000222000222AAARRR 110000220022AARR 100202AR 110000220022AAR 111000000222000222AAA 110000220022AA 110000220022AA 100202A 111000000222000222AAARR 100202AR 110000220022AARR 110000220022AARR 111000000222000222AAARRR PFOS Found (ng/mL) NNDD ND NNDD ND NNDD ND ND 3321300000 16400 331045160000000 222319560000000 28N9D00 ND NNDD NNDD ND 5N2D2 56571S1910000 82400 7813680000 722356239000000 28000 27400 N* DDu=pNlicoatteDIentjeecctteidon(Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 54 of 217 : 571 : MIN 313/023622 Exygen Study No.: 023-076 Table XXIII. Summary ofPFOA Residues in Rat Serum Samples Sponsor Exygen ID ID 1M-GROUP 1Day 92 0203367 1M-GROUP 1Day 92 0203367 223MMM---GGGRRROQOUUUPPP111DDDaaayyy999222* 00220033336688 0203369 34MM--GGRROOUUPP11DDaayy9922* 00220033336790 4M-GROUP 1Day92* 0203370 55MM--GGRROOUUPP11DDaayy9922* 6M-GROUP2 Day92 0203371 00220033337712 67MM--GGRROOUUPP22DDaayy9922 78MM--GGRROOUUPP22DDaayy9922* 8M-GROUP2 Day 92* 00220033337732 000222000333333777344 919MOMM--GG-GRRROOOUUUPPP222DDDaayayy992922* 10M-GROUP2 Day92* 1111MM--GGRROOUUPP33DDaayy9922* 1122MM--GGRROOUUPP33DDaayy9922* 00220033337755 0203376 00220033337776 0203377 00220033337788 1I33MM--GGRROOUUPP33DDaayy9922* 14M-GROUP 3 Day92 000222000333333778990 111455MMM---GGGRRROOOUUUPPP333DDDaaayyy999222** 000222000333333888110 1166NMI--GGRROOUUPP44DDaayy9922* 0203382 0203382 111787MMM---GGGRRROOOUUUPPP444DDDaaayyy999222* 18M-GROUP4 Day92* 00220033338833 00220033338844 19M-GROUP4 Day92 0203385 212090MMM---GGGRRROOOUUUPPP444DDDaaayyy999222** 0203385 00220033338866 Set Number 092002B 092002B 092002B 00992200Q022BB 00099922200000222BBB 009922000022BB 092002B 009922000022BB 009922000022BB 092002B 000999222000000222BBB 092002B 009922000022BBRR 092002BR 009922000022BBRR 092002BR 092002BR 000999222000000222BBBRRR 009922000022BBRR 092002BR 009922000022BBRR 092002BR 092002BR 092002BR 009922000022BBRR PFOA Found (ng/mL) ' 60.6 57.3 76.0 115152 777830...147 77.6 18449.50 11552400 111522825000 111113113000 331323147000 44126900 34904200 3890 223688084000 1100670000 111120197000000 10300 10600 10600 99755700 N* DDu=plNicoatteDIentjeecctteidon(Axea less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 55 of 217 : 572 : MIN 313/023622 Exygen Study No.: 023-076 Table XXm (coni'). SSuammmplaersy ofPFOA Residues in Rat Serum Sponsor ID 21F-GROUP 1Day92 21F-GROUP 1Day 92 _2222FF--GGRROOUUPP11DDaayy9922* 24F-GROUP 1Day92 2245FF--GGRROOUUPP11DDaayy9922* 2256FF--GGRROOUUPP12DDaayy9922* 222767FFF---GGGRRROOOUUUPPP222DDDaaayyy999222** 2288FF--GGRROOUUPP22DDaayy9922* 29F-GROUP 2 Day 92 233900FFF---GGGRRROOOUUUPPP222DDDaaayyy999222* 3Ir-GROUP 3 Day 92 31F-GROUP 3 Day 92* 333223FFF---GGGRRROOOUUUPPP333DDDaaayyy999222* 333344FFF---GGGRRROOOUUUPPP333DDDaaayyy999222** 3355FF--GGRROOUUPP33DDaayy9922* 333667FFF--G-GGRRROOOUUUPPP444DDDaaayyy999222* 37F-GROUP4 Day92* 3388FF--GGRROOUUPP44DDaayy9922* 39F-GROUP4 Day 92 3490FG-G3RROOUUPP44DDaayy9922* 40F-GROUP4 Day92* Exygen ro 0203387 0203387 00220033338888 0203389 0203389 000222000333333999100 00220033339912 0203392 0203393 00220033339934 00220033339954 000222000333333999566 00220033339977 0203398 0203398 00220033339999 0203400 0203400 000222000333444000112 00220033440032 00220033440034 00220033440045 0203405 Set Number 092502A 092502A 009922550022AA. 092502A 092502A 009922550022AA 092502A 009922550022AA 092502A 092502A 009922550022AA 009922550022AA 000999222555000222AAA 009922550022AA 092502A 092502A 009922550022AA 092502A 009922550022AA 092502A 009922550022AA 092502A 009922550022AA 009922550022AA 092502A PFOA Found (na/mL) ND NNDD ND NNDD ND NNDD ND NNDD NNDD ND NNDD NNDD ND NNDD ND NNNDDD ND ND 8110.27 67.6 7NN4DD.9 503 513 5533..86 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.0001 pg/mL) Exygen Research Page 56 of 217 : 573 : MIN 313/023622 Exygen Study No.: 023-076 Table XXHI (cont'). SSaummpmleasry of PFOA Residues in Rat Serum Sponsor ID Exygen ID Set Number PFOA Found (ng/mL) 41M-GROUP 1Day 121 0203406 4412MM--GGRROOUUPP11DDaayy112211* 00220033440076 42M-GROUP 1Day 121* 0203407 092502B 092502B 009922550022BB ND ND 3388.38 444334MMM---GGGRRROOOUUUPPP111DDDaaayyy111222111* 4445MM--GGRROOUUPP11DDaayy112211* 0203408 00220033440089 00220033440190 009922550022BB 092502B 009922550022BB NNDD ND ND 42.6 4456MM--GGRROOUUPP12DDaayy112211* 00220033441110 46M-GROUP2 Day 121* 0203411 009922550022BB 092502B 44487U69 4477MM--GGRROOUUPP22DDaayy112211* 00220033441122 009922550022BB 458022 48M-GROUP 2 Day 121 0203413 092502B 662 4489MM--GGRROOUUPP22DDaayy112211* 49M-GROUP2 Day 121* 000222000333444111344 009922550022BB 092502B 652758 514 555001MMM--G-GGRRROO0UUUPPP223DDDaaayyy111222111* 51M-GROUP 3 Day 121* 0000222200003333444411115566 52M-GROUP 3 Day 121 0203417 555323MMM---GGGRRROOOUUUPPP333DDDaaayyy111222111** 000222000333444111788 54M-GROUP 3 Day 121 0203419 5545MM--GGRROOUUPP33DDaayy112211* 00220033441290 009922550022BB 000999222555000222BBB 000999222555000222BBB 092502B 009922550022BB 551097 668210669 604 11116600 320994 790 55M-GROUP 3 Day 121* 56M-GROUP4 Day 121 00220033442210 5567MM--GGRROOUUPP44DDaayy112211* 00220033442212 555N6*5567890890DDMMMMMMMu---G=---GGGpGGGRRRRlNRRRiOOOOcOOOoaUUUUtUUUtPPePPDPPP4444Ie444ntDDDDDDjDeeaaaacaaacyyyytyyyte1i1111d11o22222221n1111(11****Area00000002222222l0e000000333s3333s44444442222222t3325544han 009922550022BB 092502B 009922550022BB 009922550022BB 000099992222555500002222BBBB lowest calibration standard 797004 917 11003400 553672368 377367836 of 0.0001 pg/mL) Exygen Research Page 57 of 217 : 574 : MIN 313/023622 Exygen Study No.: 023-076 Table XXIII (coni'). SSuammmplaersy of PFOA Residues in Rat Serum Sponsor Exygen ID ID 61F-GROUP 1Day 121 61F-GROUP 1Day 121* 62F-GROUP 1Day 121 62F-GROUP 1Day 121* 0203426 000222000333444222677 64F-GROUP 1Day 121 0203428 64F-GROUP 1Day 121* 0203428 666556FFF--4GG3RRROOOUUUPPP121DDDaaayyy111222111* 66F-GROUP2 Day 121* 6677FF--GGRROOUUPP22DDaayy112211* 68F-GROUP2 Day 121 68F-GROUP2 Day 121* 67679090FFFF----GGGGRRRROOQOUUUUPPPP2222DDDDaaaayyyy111122221111** 00220033442299 0203430 00220033443310 0203431 00220033443322 0000222200003333444433333344 7711FF--GGRROOUUPP33DDaayy112211* 72F-GROUP 3 Day 121 7723FF--GGRROOUUPP33DDaayy112211* 73F-GROUP 3 Day 121* 74F-GROUP3 Day 121 00220033443355 000222000333444333676 00220033443378 777545FFF---GGGRRROOOUUUPPP333DDDaaayyy111222111** 0203438 00220033443399 7766FF--GGRROOUUPP44DDaayy112211* 00220033444400 7777FF--GGRROOUUPP44DDaayy112211* 7788FF--GGRROOUUPP44DDaayy112211* 0203441 000222000333444444122 79F-GROUP4 Day 121 79F-GROUP4 Day 121* 00220033444433 8800FF--GGRROOUUPP44DDaayy112211* 00220033444444 NuSmetber 100102A 100102A 110000110022AA 100102A 100102A 111000000111000222AAA 100102A 110000110022AA 110000110022AA 100102A 110000110022AA 100102A 110000110022AA 111000000111000222AAA 110000110022AA 1001Q2A 11000011Q022AA 110000110022AA 100102A 111000000111000222AAA 110000110022AA 110000110022AA PFOA Found (ng/'mL) NET NNDD ND NNDD NNDD ND NNNDDD NNDD NNNDDD NNNDDD ND NNDD NNDD ND NNDD NNDD ND NNDD ND NNDD NNDD N* DDu=plNicoatteDIentjeecctteidon(Area less than lowest calibration standard of 0.0001 pg/inL) Exygen Research Page 58 of 217 : 575 : MIN 313/023622 Exygen Study No.: 023-076 Table XXin (cont'). SSaummpmleasry of PFOA Residues in Rat Serum Sponsor Exygen Set PFOA ID ID Number Found (ng/mL) 81M-GROUP 1Week4 8812MM--GGRROOUUPP11WWeeeekk44* 82M-GROUP 1Week4* 888334MMM---GGGRRROOOUUUPPP111WWWeeeeeekkk444* 84M-GROUP 1Week4* 88885656MMMM----GGGGRRRROOOOUUUUPPPP4141WWWWeeeeeeeekkkk4444** 0203445 000222000333444444566 00220033444477 0203448 00220033444489 0203449 00220033445500 100202A 111000G00222000222AAA 100202A 1I0000220022AA 111000000222000222AAA 110000220022AARR NO ND NNDD ND NNDD ND NNDD 76162200 8877MM--GGRROOUUPP44WWeede:k44 88888989MMMM----GGGGRRRROOOOUUUUPPPP4444WWWWeeeeeeeekkkk4444** 9900MM--GGRROOUUPP44WWeeeekk44* 000222000333444555112 0203452 00220033445533 00220033445544 111000000222000222AAARRR 111000000222000222AAARRR 110000220022AARR 33566300 7320 778782139000 99243700 9911FF-G-GRRO0UUPP11WWeeeekk44* 92F-GROUP 1Week4 9923FF--GGRROOUUPP11WWeeeekk44* 9934FF--GGRROOUUPP11WWeeeekk44 9945FF--GGRROOUUPP11WWeeeekk44* 95F-GROUP 1Week4* 96F-GROUP4 Week4 999677FFF---GGGRRRQOOUUUPPP444WWWeeeeeekkk444** 999889FFF---GGGRRROOOUUUPPP441WWWeeeeeekkk444* 919OFO-GF-RGOROUUPP1!WWeeekek4*4 0203455 00220033445556 00220033445576 0203457 00220033445588 0203459 00220033445690 00000000222222220000000033333333444444446666666611233024 110000220022AA 110000220022AA 110000220022AA 100202A 110000220022AA 110000220022AA 110000220022AA 100202A 111000000222000222AAA 110000220022AA ND NNDD NNNDDD ND ND 6N2D.6 ND NNDD ND NNNDDD NNDD N*10DD0Fu=-pGlNRicoOatUteDPIen1tjeWeccetteeikdo4n(*Area02le0s3s46t4han 100202A lowest calibration standard ND of 0.0001 pg/mL) Exygen Research Page 59 of 217 : 576 : MIN 313/023622 Exygen Study No.: 023-076 Table XXIV. Summary ofPOSF Residues in Rat Serum Samples Sponsor ID Exygen ID 11MM--GGRROOUUPP11DDaayy9922* 00220033336677 22MM--GGRROOUUPP11DDaayy9922* .33MM--GGRROOUUPP.11DDaayy9922* 44MM--GGRROOUUPP11DDaayy9922* 000222000333333666889 000222000333333677900 S6785768MMMMMMMM--------GGGGGGGGRRRRRRRROOOOOOOOUUUUUUUUPPPPPPPP22212221DDDDDDDDaaaaaaaayyyyyyyy9999999922222222**** 00000000222222220000000033333333333333337777777711223344 919M0MM--GG-RGROROUOUPUP22PD2DDaayayy999222* 000222000333333777556 111011MMM---GGGRRROOOUUUPPP233DDDaaayyy999222** 000222000333333777767 12M-GROUP3 Day92 11112334MMMM----GGGGRRRROOOOUUUUPPPP3333DDDDaaaayyyy99992222** 14M-GROUP 3 Day92* 00220033337788 00220033337799 00220033338800 1155MM--GGRROOUUPP33DDaayy9922 111667MMM---GGGRRROOOUUUPPP444DDDaaayyy999222* 111788MMM---GGGRRROOOUUUPPP444DDDaaayyy999222** 00220033338811 00220033338822 0000222200003333333388883344 1199MM--GGRROOUUPP44DDaayy9922* 00220033338855 2200MM--GGRROOUUPP44DDaayy9922* 00220033338866 Set Number 110000110022AA 100102A 111100000000111100002222AAAA. 100102A 111000000111000222AAA 110000110022AA 111000000111000222AAA 111000000111000222AAA 019020610022BA 092602B 009922660022BB 000999222666O00222BBB 009922660022BB 092602B 000999222666000222BBB 000999222666000222BBB 000099992222666600002222BBBB POSF Found (ue/mL) ' NNDD NNNDDD NNDD NNDD NNDD NNDD NNNDDD NNNDDD NNDD NNDD ND NNNDDD NNNDDD NNNDDD NNNDDD NNDD NNDD N* DDu=pNlicoatteDAetleiqctueodt (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 60 of 217 : 577 : MIN 313/023622 Exygen Study No.: 023-076 Table XXIV (coni'). SSaummpmleasry of POSF Residues in Rat Serum Sponsor ID 2211FF--GGRROOUUPP11DDaayy9922* 2222FF--GGRROOUUPP11DDaayy9922* 2244FF--GGRROOUUPP11DDaayy9922* 222556FFF---GGGRRROOOUUUPPP121DDDaaayyy999222* 26F-GROUP 2 Day 92* 2277FF--GGRROOUUPP22DDaayy9922* 2288FF--GGRROOUUPP22DDaayy9922* 2299FF-G-GRROOUUPP22DDaayy9922* 33330101FFFF---GG-GGRRRROOOOUUUUPPPP2323DDDDaaaayyyy99992222** 3322FF--GGRROOUUPP33DDaayy9922* 33333434FFFF---GGGGRRRROOOOUUUUPPPP3333DDDDaaaayyyy99992222** 3355FF--GGRROOUUPP33DDaayy9922* 3366FF--GGRROOUUPP44DDaayy9922* 33337878FFFF-G--GGGRRRROOOOUUUUPPPP4444DDDDaaaayyyy99992222** 34349009FFFF-G--GGGRRRROOOOUUUUPPPP4444DDDDaaaayyyy99992222** Exygen m 0203387 0203387 00220033338888 00220033338899 00220033339900 0203391 000222000333333999122 00220033339933 00220033339944 000000000222222222000000000333333333333333333999999999S56677889 000222000333344900900 0203401 00220033440012 000222000333444000332 000222000333444000544 0203405 Set Number 092702A 009922770022AA 092702A009922770022AA 000999222777000222AAA 000999222777000222AAA 009922770022AA 009922770022AA 000099992222777700002222AAAA 1110Q0000222000222AAA 110000220022AA 100202A 110000220022AA 110000220022AA 111000000222000222AAA 100202A 111100000000222200002222AAAA . ... . POSF Found (jig/mL) ND ND NNNDDD ND NNNDDD ND NNDD NNDD NNNDDD NNNDDD NNNDDD NNDD ND NNDD ND NNDD NNNDDD NNDD NNDD * Duplicate Aliquot ND = Not Detected (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 61 of 217 : 578 : MIN 313/023622 Exygen Study No.: 023-076 Table XXIV (cont'>. SSaummpmleasry ofPOSF Residues in Rat Serum Sponsor Exygen ID ID 44I2MM--GGRROOUUPP 11DDaayy 112211 42M-GROUP 1Day 121* -43M-GROUP1Day 121 4434MM--GGRROOUUPP11DDaayy112211* 0203406 000222000333444000778 0203408 0203409 444455MMM---GGGRRROOOUUUPPP111DDDaaayyy111222111** 000222000333444011900 46M-GROUP2 Day 121 4444467878MMMMM-----GGGGGRRRRROOOOOUUUUUPPPPP22222DDDDDaaaaayyyyy111112222211111*** 000222000333444111112 000222000333444111233 45459090MMMM----GGGGRRRROOOOUUUUPPPP2222DDDDaaaayyyy111122221111** 5511MM--GGRROOUUPP33DDaayy112211* 5522MM--GGRROOUUPP33DDaayy112211* 00220033441144 000222000333444111556 000222000333444111677 555334MMM---GGGRRROOOUUUPPP333DDDaaayyy111222111* 000222000333444111889 54M-GROUP 3 Day 121* 555S5555675678MMMMMMM------G-GGGGGGRRRRRRROOOOOOOUUUUUUUPPPPPPP4434344DDDDDDDaaaaaaayyyyyyy111111122222221111111*** 00220033441290 000222000333444222110 000222000333444222322 5565689090MMMMM-----GGGGQRRRRROOOOOUUUUUPPPPP44444DDDDDaaaaayyyyy111112222211111*** 000222000333444222344 00220033442255 NuSmetber 110000330022AA 110000330022AA110000330022AA 100302A 111100000000333300002222AAAA 111100000000333300002222AAAA 100302A 110000330022AA 110000340022AA 110000440022AA 100402A 111000000444000222AAA 111000000444000222AAA 110000440022AA 111111110000000000000000444444440000000022222222AAAAAAAA POSF Found (ue/mL) NNDD' NNDD NNDD ND NNDD NNNDDD NNDD ND NNNNNDDDDD NNNDDD NNNDDD NNNDDD NNNNNDDDDD NNDD NNNDDD N* DDu=pNlicoatteDAetleicqtueodt(Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 62 of 217 : 579 : MIN 313/023622 Exygen Study No.: 023-076 Table XXTV (cont'). SSaummpmleasry of POSF Residues in Rat Serum Sponsor ID Exygen ID 6IF-GROUP 1Day 121 0203426 6612FF--GGRROOUUPP11DDaayy112211* 00220033442276 62F-GROUP1Day 121* 0203427 6644FFGGRROOUUPP11DDaayy112211* 00220033442288 666S65FFF--GGGRRROOOUUUPPP121DDDaaayyy111222111* 66666778FFFF--GGGGRRRROOOOUUUUPPPP2222DDDDaaaayyyy111122221111** 68F-GROUP2 Day 121* 000000222222000000333333444444322333191900 00220033443322 76670990FFFF----GGGGRRRROOQOUUUUPPPP2222DDDDaaaayyyy111122221111** 000222000333444333334 0203434 7711FF-GGRROOUUPP33DDaayy112211* 00220033443355 777223FFF--GGGRRROOOUUUPPP333DDDaaayyy111222111* 000222000333444333766 777345FFF--GGGRRROOOUUUPPP333DDDaaayyy111222111* 7756FF--GGRROOUUPP34DDaayy112211* 7767FF-GGRROOUUPP44DDaayy112211* 77FGROUP4Day 121* 0203437 000222000333444333899 0000222200003333444444441100 77877889089QFFFFFF-----GGG-GGGRRRRRROOOOOOUUUUUUPPPPPP444444DDDDDDaaaaaayyyyyy111111222222111111*** 000222000333444444232 000222000333444444344 Set Number 100402A 100402A 100402A 111Q0000Q44400G222AAA' 110000440022AA 111000O00444O00222AAA 100402A 111110000000000444440000022222AAAAA 111000000444000222AAA 100702A 110000770022AA . 111000000777000222AAA 110000770022AA 111000000777000222AAA 110000770022AA 100702A 111000000777000222AAA , POSF Found (ug/mL) ND NNDD NNDD ND NNNNNNDDDDDD NNDD NNNDDD NNDD ND NNNDDD NNNDDD NNDD NNNDDD NNNDDD NNNDDD N* DDu=pNlicoatteDAetleiqctueodt (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 63 of217 : 580 : MIN 313/023622 Exygen Study No.: 023-076 Table XXTV (coni'). SSaummpmleasry ofPOSF Residues in Rat Serum SpoIDnsor Exygen ID 81M-GROUP 1Week 4 0203445 8822MM--GGRROOUUPP11WWeeeekk44* 00220033444466 "8843MM--GGRROOUUPP11 WWeeeekk 44 84M-GROUP1Week4* 8885S6MMM---GGGRRROOOUUUPPP141WWWeeeeeekkk444* 000222000333444444788 0203449 00220033444590 888677MMM---GGGRRROOOUUUPPP444WWWeeeeeekkk444** 00220033445510 0203451 8888MM--GGRROOUUPP44WWeeeekk44* 00220033445522 989090MMM---GGGRRROOOUUUPPP444WWWeeeeeekkk444* 000222000333444555344 999121FPF--G-GGRRROOOUUUPPP111WWWeeeeeekkk444* 000222000333444555556 9923FF--GGRROOUUPP11WWeeeekk44* 00220033445576 9934FF--GGRROOUUPP11WWeeeekk44* 9945FF--GGRROOUUPP11WWeeeekk44* 999566FFF---GGGRRROOOUUUPPP414WWWeeeeeekkk444** 0203457 000222000333444555889 0Q0222000333444566900 9977FF--GGRROOUUPP44WWeeeekk44* 00220033446611 999889FFF---GGGRRROOOUUUPPP441WWWeeeeeekkk444 000222000333444666322 91O9FO-FG-RGOROUUPP11WWeeekek4*4 00220033446634 100F-GROUP1Week4* 0203464 NuSmetber 101702A 101702A 101702A 111000111777000222AAA111000111777000222AAA 111000111777000222AAA 101702A 110011770022AA 111110000011000787880000022222AAAAA 110000880022AA 110000880022AA 110000880022AA 110000880022AA 100802A 110000880022AA 111000000888000222AAA 110000880022AA 100802A . POSF Found (iis/mL) ND NNDD ND NNDD NNNDDD NNNDDD NNDD NNDD NNNNDDDD NNNDDD ND NNDD NNDD NNDD ND NNNDDD NNDD ND N* DDu=pNlicoatteDAetleicqtueodt (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 64 of 217 : 581 : MIN 313/023622 Exygen Study No.: 023-076 Table XXV. Summary ofPFOS Residues in Rat Urine Samples Sponsor ID Exygen ID Set Number PFOS Found (ng/mL) NN*DDQu==plNNicooattteDQ21U1111111124536791821111111111I24735689e10u023456789***n****10537802469*t*******a*jeenccttitefiidoanb(Alere(Raelesspso0000000000000000000000000000000000000000tn2222222222222222222222222222222222222222hs0000000000000000000000000000000000000000ae3333333333333333333333333333333333333333n5555555555555555555555555555555555555555l66667667666666667777877777777778777788o88t33514h5714678899351604022345771668899w0202aenstthcealairbera11a11111111111111111111111111111111111111t000000000000000000o00i00000000000000000000000000000000000000of0000000000000000000000333333333333333333n3t333333333333333333333000000000000000000hQ00000000000000s0000002222222222222222202et222222222222222222222AAAAAAAAAAAAaAAAAAA2AAAAAAAlAAAAAAAAAAAAAAnoARRRRRRRRRRRRRRRRRRdwa'ersdtocfal0i.b'0r0a0ti'1onpgst/33a1111m1NNNNNNNNNN442453311111891111119118n37400015068951225108966392293DDDLDDDDDQQ7d399607119337304778930082500490)a000000rd but less than the LOQ. Exygen Research Page 65 of 217 : 582 : MIN 313/023622 Exygen Study No.: 023-076 Table XXV (coni'). Summary of PFOS Residues in Rat Urine Samples Sponsor ID Exygen .. ID Set Number PFOS 2211* 2222* 23 2234* 24* 2255 26 2267* 222878** 2299* 3300* 31 333122* 3333* 34 34* 35 333656** 333778* 3389* 434090** 0203583 00220033558834 0203584 0203585 00220033558856 00220033558867 0203587 00220033558888 0203589 00220033558990 0203590 00220033559911 0203592 00220033559932 00220033559934 0203594 0203595 0203595 00220033559966 0203597 0203597 00220033559988 00220033559999 000222000333666000100 000222000333666000122 110000770022AA 111000000777000222AAA. 110000770022AA I00702A 110000770022AA 100702A 110000770022AA 100702A 100702A 100702A 110000770022AA 111000000777000222AAA 111000000777000222AAA 110000770022AA 110000770022AA 100702AR 100702AR 110000770022AARR 110000770022AARR 110000770022AARR 100702AR 111000000777000222AAARRR NNDD ND ND NNNDQD NQ NQ NQ 49.1 5134.00 3162.87 30.5 2249..27 15.8 1741.91 671 390 458372 577 441212 238 255 21094700 11874200 44174100 1540 111365848000 NN* DQDu==pNNlicooattteDQIeuntajeenccttietfiodianb(Aler(eRaelesspsotnhsaen5lothwaenstthcealairberaatoiofnthsetalnodwaerdstocafl0i.b0r0a0ti1onpgst/amnLda)rd but less than the LOQ. Exygen Research page 66 of 217 : 583 : MIN 313/023622 Exygen Study No.: 023-076 Table XXV (cont'). Summary ofPFOS Residues in Rat Urine Samples Sponsor Q) 41 41* 4422* 43 43* 44 44* 4455* 4466* 47 47* 4488* 4499* 555010* 555122* 5533 54 5545* 55* 5566* 5577* 58 58* 59 59 6600* Exygen .. ID 0203603 0203603 00220033660044 0203605 00220033660056 000222000333666000776 000222000333666000898 00220033660190 00220033661110 0203611 0203612 00220033661132 00220033661134 0203614 00220033661155 00220033661166 0203617 00220033661178 0203618 00220033661199 00220033662200 0203621 000222000333666222122 Set Number 100702B 100702B 110000770022BB 100702B100702B 100702B 100702B 110000770022BB 1007020 11000077Q022BB 110000770022BB 110000770022BB 10O7Q2B 110000770022BB 100702B 111000000777000222BBB 110000770022BB 101402B 11001144O022BB 111000001774000222BBBRR 110000770022BBRR 110000770022BBRR 100702BR 111000000777000222BBBRRR PFOS ND NDNNDD NNDD ND ND NNDD 4533361975.....78707 4558..52 4X8 70.1 6175.88 911405.795 240 232 430229 253 924950 895 11126200 883 1825510 1552323060 NNDQ== NNoottDQeutaencttiefdiab(Alere(Raelesspsotnhsaenlothwaenstthcealaibreraatoiofnthsetalnodwaerdstocfal0ib.0r0a0ti1on(igst/amnLda)rd but less than theLOQ. Exygen Research Page 67 of 217 : 584 : MIN 313/023622 Exygen Study No.: 023-076 Table XXV (cont'). Summary of PFOS Residues in Rat Urine Sample Sponsor ED Exygen Set ID Number PFOS 61 61* 62 6624* 64* 6655* 66 66* 6677* 68 666899** 70 7701* 71* 72 72 73 73* 74 74* 75 75* 76 777767** 777898* 788900** *Duplicate Injection 00220033662233 0203624 0203624 0203625 00220033662256 0203626 00220033662277 0203628 0203628 0203629 00220033662390 0203630 00220033663311 0203632 00220033663332 0203633 00220033663344 0203635 00220033663356 00220033663376 0203637 0203638 0203638 0203639 00220033663490 00220033664410 0203641 102102B 102102B 102102B 110022110022BB . 102102B 102102B 110022110022BB 102102B 102102B 110022110022BB 111000222111000222BBB 102102B 102102B 110022110022BB 102102B 102102B 110022110022BB 1I0022110022BB 111000222111000222BBB 102102B 102102BR 102102BR I1002211O022BB 102102BR 110022110022BBR 102102B ' ND ND ND NNDD NNDD ND 7668..19 32.9 33.5 65.1 56321.82 441776..138 211512537 221 332580 132 112752 512668 473 1580 1450 630 263750 277464 674 ND = Not Detected (Area less than lowest calibration standard of0.0001 pg/mL) NQ = Not Quantifiable (Response than the area of the lowest calibration standard but less than the LOQ. Exygen Research Page 68 of217 : 585 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVI. Summary ofPFOA Residues in Rat Urine Samples Sponsor m Exygen ID Set Number PFOA 11* 22* 3 434** 55 66 7 788* 9 9* 1100* 11 1112* 12* 1111I43S34** I1S6* 16* 17 1178* 18 19 19* *Duplicate22I0n0*jection 0203563 0203563 0203S64 00220033556654 0203565 0203566 000222000333555666767 000222000333555666889 0203569 0203570 0203570 0203571 00220033557712 00220033557732 0203573 00220033557744 0Q0222000333555777556 00220033557767 000222000333555777878 00220033557799 0203580 0203580 0203581 000222000333555888122 110000330022AA 100302A 100302A 100302A. 100302A 100302A 110000330022AA 110000330022AAR 110000330022AAR 110000330022AAR 100302AR 110000330022AARR 110000330022AARR 100302AR 110000330022AARR 100302AR 110000330022AARR 110000330022AARR 100302AR 111000000333000222AAARRR 100302AR 100302AR 100302AR 110000330022AARR 111000000333000222AAARRR ' NNQD NNQQ 14.8 1N4Q2 NNNQQQ 203 317979 383 237 222343800 253682384 552684 582 778876 446530 711 55741204800 44343000 2060 2230 2700 222633941000 NNQD =~ NNoott DQeutaencttiefidab(Alere(Raelesspsotnhsaen>lothwaenstdciealaibreraatoiofnthsetalnodwaersdtocfal0i.b0r0a0ti1onpgst/amnLd)ard but less the LOQ Exygen Research Page 69 of 217 : 586 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVI (coni'). Summary ofPFOA Residues in Rat Urine Samples Sponsor ID 21 21 22 2223 2234* 24* 2255* 26 2267* 222788** 2299* 333010* 333122* 333334* 34* 35 35* 3366* 3377* 38 333989** 4400* Exygen ID 00220033558833 0203584 0203584 0203585 00220033558856 000222000333555888776 00220033558888 0203589 00220033558990 0203590 00220033559911 000222000333555999322 000222000333555999344 000222000333555999556 0203596 0203597 00220033559978 0203598 0203599 00220033569090 00220033660010 000222000333666000122 Set Number 100702A 100702A 100702A 111100000000777700002222AAAA . 100702A 110000770022AA 100702AR 100702AR 100702A 110000770022AA 100702A 100702AR 100702AR 100702A 110000770022AAR 100702AR 110000770022AARR 100702AR 100702AR 100702AR 100702AR 100702AR 110000770022AARR 100702AR 100702AR 100702AR 100702AR 100702AR 100702AR 111000000777000222AAARRR PFOA ' 13.4 12.4 NNNQQD ND NQ NQ 1145..14 358 435732 413849 192 226796 116676 2450 221409115000 22559500 11897100 1300 71926800 8000 1177290000 18100 1180630000 111012480000000 NN* DQDu==pNNlicooattteDQieuntajeenccttitefiodianb(Alere(Raelesspsotnhsaenlothwaenstthcealaibreraatoiofnthsetalnodwaerdstocaf0li.b0r0a0ti1onpgst/amnLda)rd but less than theLOQ Exygen Research Page 70 of 217 : 587 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVI (cont'). Summary of PFOA Residues in Rat Urine Samples Sponsor ID Exygen ID Set Number PFOA 41 4412* 4423* 444344** 45 4456* 46* 47 4478* 48* 445990* 555011** 5522* 555343* 54* 5555* 56 5567* 5578* 555899** * Duplicate66I0n0*jection 0203603 0203603 0203604 00220033660045 00220033660056 0203606 00220033660077 0203608 00220033660089 0203609 00220033661100 0203611 00220033661112 0203612 00220033661133 00220033661144 000222000333666111556 000222000333666111767 0203618 00220033661189 00220033661290 00220033662210 0203621 00220033662222 100702B 100702B 10Q702B 110000770022BB 100702B ' 110000770022BB 110000770022BB 110000770022BB 100702B 110000770022BB 100702B 111000000777000222BBB 1100C0770022BB 111000000777000222BBB 111000000777000222BBB I00702B 110011440022BB 100702BR 100702BR 111000000777000222BBB 100702B 110000770022BB 110000770022BB * NNDD ND NNDD ND NNDD ND ND 478360...170 444584...117 444090...390 4793..84 69.8 8803..15 115574 53.7 51646.60 1740 241 203 111436873 111171156 192 189 NNDQ =~ NNoott DQeutaencttiefdia(bAlere(Raelesspsotnhsaen lothwaenstdciealairberaatoiofnthsetalnodwaersdtocafl0i.b0r0a0ti1onpgst/amnLda)rd but less than the LOQ Exygen Research Page 71 of 217 : 588 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVI (cont'). Summary ofPFOA Residues in Rat Urine Samples Sponsor ID Exygen ID Set Number PFOA 61 61* 62 62* 64 6645* 65* 66 66* 67 67* 68 68* 69 69* 70 7701* 71* 7722 7733* 74 74* 75 75* 76 76* 77 77* 78 78* 79 79* 80 80* * Duplicate Injection 0203623 0203623 0203624 00220033662254 00220033662256 00220033662267 0203627 0203628 00220033662289 0203629 00220033663300 00220033663311 0203632 00220033663332 00220033663343 0203634 00220033663355 0203636 00220033663376 00220033663387 0203638 000222000333666343909 0203640 0203641 0203641 110022110022BB 111000222111000222BBB_ 110022110022BB 102102B 110022110022BB 110022110022BB 102102B 111000222111000222BBB 102102B 102102B 1100221I0022BB 102102B 110022110022BB 102102B 110022110022BB 110022110022BB I02102B 102102B 110022110022BB 111000222111000222BBB 110022110022BB 102102B ' NNDD NNDD ND NNDD NNNDDD NNDD ND NNDD ND NNDD NNNDDD NNDD 233 NNDD NQ NQ 19.6 20.8 23.4 2N3D.3 1N2D3 11.9 ND ND NNDQ ==NNoott DQeutaencttiefdia(bAlere(Ra elesspsotnhsaen5lothwaenstthcealaibreraatoiofnthsetalnodwaerdstocfal0i.b0r0a0ti1onjigst/amnLd)ard but less than the LOQ Exygen Research Page 72 of217 589 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVII. Summary of POSF Residues in Rat Urine Samples Sponsor ED 1 1* 323.. 545* 66* 98978** 1110 1112* 111323** 1144* 111556* 1178 1189* 122910* 22224234* 2255* Exygen n> 00000222220000033333555556666635534 00220033556676 0203567 00220033556688 000222000333555677900 000000222222000000333333555555777777113324 00220033557754 000222000333555777566 00220033557777 00220033557789 00220033558800 000222000333555888112 00220033558834 0203585 00220033558866 00220033558877 Set Number 110011000022AA 101002A 101002A 111000111900000222AAA . 101002A 110011000022AA 111000111000000222AAA 111000111000000222AAA 111000111111000222AAA 111110000011111111110000022222AAAAA 101102A 110011110022AA 11100011I111000222AAA 110011110022AA 111000111144000222AAA 1110001I1444O00222AAA 110011440022AA POSF Found (pg/mL) ' ND ND NNDD ND NNNDDD NNDD NNNDDD NNNDDD NNNDDD NNDD NNDD NNNNDDDD NNNDDD NNDD NNNDDD NNDD NNNDDD N* DDu=plNicoatteDAetleicqtueodt(Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 73 0f 217 : 590 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVII (coni'). Summary of POSF Residues in Rat Urine Samples Spnon>sor ExIyDgen Set Number POSF Found (iig/mL) 26 22228789* 3300* 3312 3334 3356 4433317890 4422* 44444334** 45 4466* 444444798978*** s5o0* 5511* * Duplic5a55532t32e**Aliquot 00220033558889 0203590 00000222220000033333555559999913022 00220033559954 00000222220000033333555659990978906 0203601 000222000333666000342 00220033660045 000222000333666000566 0203607 00220033660088 000222000333666001990 0203610 000222000333666111112 0203612 000000222222000000333333666666111111335544 101402A 101402A 111000111444000222AAA110011440022AA 1022Q2A 110022220022AA 111000222222000222AAA 111000222222Q00222AAA 111000211255000222AAA 110011550022AA 111000111555000222AAA 111000111555000222AAA 101502A 110011550022AA 111000111555000222AAA 110011550022AA 111000222999000222AAA 111000222999000222AAA ' NNDO NNDD NNDD ND NNNDDD NNNDDD NNNNNNDDDDDD NNNDDD NNDD NNNDDD NNNNNDDDDD NNNDDD NNDD NNNNDDDD ND = Not Detected (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 74 of 217 : 591 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVII (cont'). Summary of POSF Residues in Rat Urine Samples Sponsor ID ExIyDgen Set Number POSF Found Pis/mLl 54 54* 555556* 56* 5577* 5588* 556990* 666011** 666224* 6645* 666566** 666787* 666899** 70 7701* 7722* 777434* 00220033661166 000222000333666111778 000222000333666111899 00220033662200 000222000333666222112 0203622 00000222220000033333666662222253344 00220033662256 0000222200003333666622228776 00220033662289 000222000333666233900 00220033663311 000222000333666333332 000222000333666333554 102902A 1111000022229999Q0002222AAAA110022990022AA 1Q29Q2A 110022990022AA 111100002222999900002222AAAA 110022990022AA 111000122699000222AAA 110011660022AA 111111000000111111666666000000222222AAAAAA 110011660022AA 101602A 110011660022AA 111000111666000222AAA 111000221116000222AAA NNDD NNDD ND NNDD NNDD ND NNNDDD NNNDDD NNNDDD NNDD NNNDDD NNDD NNDD NNDD NNDD NNNNDDDD NNDD * Duplicate Aliquot ND = Not Detected (Area less than lowest calibration standard of 25 pg/mL) Exygen Research Page 75 of 217 : 592 : MIN 313/023622 Exygen Study No.: 023-076 Table XXVII (coni'). SSaummpmleasry of POSF Residues in Rat Urine Sponsor m 75 75* 76 777778* 777899** 8800* Exygen ID 0203636 00220033663376 00220033663388 0203639 000222000333666344900 00220033664411 Set Number 102102A 110022110022AA 102102A110022110022AA 111000222111Q00222AAA 110022110022AA . POSF Found (us/mL) ND ND ND NNNDDD NNNDDD NNDD * Duplicate Aliquot ND = Not Detected (Area less than lowest calibration standard of 25 (ig/mT.) Exygen Research : 593 : Page 76 of 217 MIN 313/023622 Exygen Study No.: 023-076 FIGURES Exygen Research : 594 : Page 77 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 1. Typical Calibration Curve for PFOS * 100102A.rdb (PRO S): T in e a s Regression (*11 weighting): y 1084.56 x + 75.115 ( r 0.9S78965) Exygen Research : 595 : Page 78 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 2. Typical Calibration Curve for PFOA 100102A.KS) (PFO A): lin ear' Regression ("1 / x ' weighing): y 18602.7 x + 390.509 (r - 0.9972473) Exygen Research : 596 : Page 79 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 3. Typical Calibration Curve for POSF MVO/F'Qjrai^qptilrsateuereplreialrnifttenctoMdifKrnobaenteher::::::: 3L10p6LE:1o0G:\\0s102of203c30-St-C02t72d7-6065.\0\,RW2AS5UXptCuiO9dAAo:yT5#TfA30A\S21\m1O0301-O100uu1f0g06s2/i2AanAL\gP\,HOA10S*c1FLq1.M0Mh2ee0tah5doJ)dspPaAceLS02 RcP_oi_etnt_fc_Te_ainnn_tder_a:Ttiyopoe:: 34P1O.95S32F.33RpTgED/aiTi used Signal Ratios Liarits RT U nits m261099.o...000o000 1 x42010..o..501x 1806...001--- 63041...308 3333....S555Z222 t33o..60 Rasp mteg Type 41M0925S91I7873580 tissyyyysssj ddd*eexffft Exygen Research : 597 : Page 80 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 4. PCFhOroSmaantdogPrFamOARepresenting a 0.1 ng/mL standard for Exygen Research : 598 : Page 81 of 217 MIN 313/023622 Exygen Study No.: 023-076 FPOigSuFre 5. Chromatogram Representing a 25 pg/mL standard for "VAFOI.s.najic.ip*msla.qeet?.lurpr.uialHeIrmtneoud!nerm?anbmteer::: L:\023-076\hAhDATA\101102AVW110201.0 1pC23o1E1wGGs0oCf1. //WcMrstutS2d-21o80.>025StpuLd9yro0#f2022a35m-07u/ 6s i.n, g Accpethod PALS02 ImL headspace Exygen Research : 599 : Page 82 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 6. PCFhOroSmaantdogPrFaOmAR(eEpxreysgeenntiInDg0a2C01o6n8tr4oCl RonattrLolivAe,rSSeatm: ple for Exygen Research : 600 : Page 83 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 7. fCohrrPoOmSaFtog(EraxmygeRnepIDre:s0e2n0ti2n8g7a7,CSoetn:tr1o0l2R50a2tAL)iver Sample oAMInpcsleqterruauitrmoeder nt ;::: L2c5a:g\x0cO2/3cmt-0s270S60\R2AM1U0:T0A2\1iW02CSu0*s2inAWg\HA02o*Sp0w*2t0b7o.d0* wu.502 * a w V ial Muaferr: 4 J Exygen Research : 601 : Page 84 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 8. Chromatogram Representing a Control Rat Serum Sample f0o9r25P0F2OAS) and PFOA, (Exygen ID: 0201682 Control A, Set: Exygen Research : 602 : Page 85 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 9. fCohrrPoOmSaFto,g(rEaxmygRenepIrDe:se0n2t0in1g68a2CCotnrlt,rSoeltR: a0t9S26er0u2mB)Sample APOVSHI na1icpims3laqeeCtlurprualiHeirmtneoudNfermonabmteer:!;:::: LoC2S8a:6\tko0UG2nSCd3te/-ryM"p0o#S7l026, 2S0\R30ImM2~LWIDhAt3ernT:a3.dA8s\pHcpaoSmcx2e.6u0s2i0n7g.DA cqM ethod ^PAL50.2 r,.l ,i.r - o * x (^XT' Exygen Research : 603 : Page 86 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 10. fC1o0hr0rP3o0Fm2OAaSt)oagnradmPFROepAr,e(sEenxytignegnaIDC:on02tr0o1l6R83atCUornintreoSl Aam, Spelet: Exygen Research : 604 : Page 87 of217 MIN 313/023622 Exygen Study No.: 023-076 Figure 11. fCohrrPoOmSaFto,g(rEaxmygRenepIrDe:se0n2t0in2g87a5CCotnrlt,rSoletR: a1t0U11r0in2Ae S)ample vOAsMIFnaiiicpansslqeteclprurulaieNirmtneoidfNmeroanbmte::er::::: L381Sc6:1\ot60ugn2doc3t/ycr-m#0ot70sl622.\083J01U-02Wm76_O1Ah0eT:1aAd9\1as0pn1a1uQcesZi>Anog\HtoAOz.cllq*0M-2,0*e,7th.DodCWpLal5a0-2q - c i Exygen Research : 605 : Page 88 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 12. w(CEhitxrhyog1me0nantIogDg/g:ra0om2f0PR1F6eO8p4SreSaspnekndtAPinF,gSOCeAto:n1t0r0o8l0R2aBt)Liver fortified Exygen Research : 606 : Page 89 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 13. Chromatogram Representing Control Rat Liver fortified at 510p2g5/0g2wAi)th POSF (Exygen ID: 0202877 Spk A Set: iVoAisFnnapicisasaeqltcpl.rruualieNirtmneoudfMremonabtneer::::::: LF1c2S5a0p:okG\i0rkOCt2/icM3ftAi-Sc02,7a0860tSi\2oWjinL>1A0otnT:f1A5cUMotfatnc0tnTu2tr5Uiu_ot0_rsl.2iu.An1kgS\/a-tu0A>td2c+y5hq0#Me402ae20dt3h-.soDQpda7c6PeALS02 Exygen Research : 607 : Page 90 of217 MIN 313/023622 Exygen Study No.: 023-076 Figure 14. wSCphitkrhoA1m,0aSnteogtg/:mr0a1Lm0o0R2f0Pe2pFArOe)SseanntidngPFCOonAtr(oElxRyagtenSeIrDu:m02fo0r1t6i8fi2ed Exygen Research : 608 : Page 91 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 15. wCihthro5mpagt/orgnrTa.mofRPeOpSrFese(EntxiynggenCoIDnt:r0o2l 0R1a6t8S2eSrupmk Afo,rSteifti:ed 092602B) AFOSMVI naicpiimlssaqeectlpurraliuNeIrtnemoudNfrmoeabmneters:::::: Lo2SC9a:6\pok0Gin2kSC3teer/-pMo0A7lS26, 80\PR0SoA2prLWt ioDl i3fAc:5Ia1A10t i0\Hop1mnS92,/m6u0sSL2it,nu08dgIm.yD#AL0c2hq3Me-afedrtfhsrpeo d PAL-5-02 aoc:ev - ? n-w w . -- Exygen Research : 609 : Page 92 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 16. wCihtrho1m0antogg/mraLmoRf PepFrOesSenantidngPFCOonAtr(oElxRyagtenUIrDin:e0f2o0r1ti6fi8e3d Exygen Research 610 : Page 93 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 17. 5C10ph1gr1o/0mm2LAat)wogitrhamPORSeFpr(EesxeyngtienngECDo:n0t2r0o2l8R7a5tSUprkinAe,fSoertt:ified at Exygen Research : 611 : Page 94 of217 MIN 313/023622 Exygen Study No.: 023-076 Figure 18. aC1n0h2dr1oP0mF2AOat)Aog(rEamxygReenpIrDes:e0n2ti0n3g55R3a,tSLpiovnesroSraImD:pl9e1foSretP:FOS Exygen Research : 612 : Page 95 of217 MIN 313/023622 Exygen Study No.: 023-076 Figure 19. (CEhxryogmenatIoDg:ra0m20R35e1p5r,eSsepnotninsogrRIaDt:L5i2v,eSreSt:am10p2l5e0f2oAr)POSF FoAMviSnpiaiclsalessqtelcprruualiiKertmnoeBdfNreonbamteer::::::: Lc2Ss1a*53t:\ukga0doc>2y/c3t#ke-t00s72260832\0-0R023A756K11O50C,:5T31AUlaL025ub0su2indag\WsApJ2accSqeM02eUth.oDd PAL502 Exygen Research 613 : Page 96 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 20. aCnhdroPmFOatAog(rEamxygReenpIrDes:e0n2ti0n3g4R50a,tSSpeornusmorSIaDm:p8l6eMfo-r PFOS Exygen Research : 614 : Page 97 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 21. (SCEehxtr:yo0gm9e2na6tIo0Dg2:rBa0)m20R33e7p7r,eSsepnotninsogrRIaDt:SHerMum-GSraomuppl3e DfoaryP9O2S, F - VOAWPSI naicpiismlasqeetelruprualiNIermtneouNdfemronabmteer::::::: LcS2Sa6:at\ukGm0dSC2pye/3Mlp#-e0S,07226#083\00R-22wA0oa3-n3{4A7n:7TU2,A9 \HS260211.D pm u s in g AcqMethod l^mL headspace 12 PALS02 Exygen Research : 615 : Page 98 of 217 MIN 313/023622 Exygen Study No.: 023-076 Figure 22. aCnhdroPmFOatAog(rEamxygReenpIrDes:e0n2ti0n3g5R73a,tSUproinnseoSraImD:p1le1fSoertP: FOS Exygen Research : 616 : Page 99 of217 MIN 313/023622 Exygen Study No.: 023-076 Figure 23. Chromatogram Representing Rat Urine Sample for POSF (Exygen ID: 0203573, Sponsor ID: 11, Set: 101102A) AmOIswvnaicpimsasqetelrpurualiiNermtneoudMfermonubtseer: 1JLS1_Ea21:_tmG\uG0_OdC2p/y3clMe-t0S27J2N36082\-00R023A756M71D31A,:4T11A\a1Ls0Xhu1es0ai2ndAg\sHpA0ac1cq1e>0t2etUho.Dd pal502 Exygen Research : 617 Page 100 of 217 MIN 313/023622 Exygen Study No.: 0234)76 APPENDIX A (ExygeSntuSMdtuyIdNPy/r3No1to3o.c0o2l 3-076) Amendmentsanadnd Deviations Exygen Research : 618 : Page 101 of 217 Study Number : MDOT13 CONFIDENTIAL MIN 313/023622 Exygen Study No.: 023-076 r -u c i'H Huntingdon ' Life Sciences PROTOCOL PERELUOROOCIANESULroNYI,FLUORIDE(POST; T-766L4) TOXICITYSTUDYBY INHALATIONADMINISTRATIONTO CDRATSFOR13WEEKS FOLLOWEDBYA4-WEEKRECOVERYPERIOD 33MMCCeonrpteorrateTtmcolegy SBttnPitafidng 22C-2E-02 USAMN 55133-3220 WnnfiJ V>n Ts fm WAkooacObetytrRyood CPEam2GShLr4iAHrfNgSBDahnc T TcoJ amber cfpegca 25 Final Protocol Page Exygen Research : 619 Page 102 of 217 MIN 313/023622 Exygeo Study No.: 023-076 SlndyNumber : MTN/313 CONTACTDETAILS Huntingdon * Life Sciences Sponsor*i Montana* Sciata Z- ' Exygen Research : 620 : Page 103 of 217 MIN 313/023622 Exygen Study No.: 023-076 Huntingdon Life Sciences PROTOCOL APPROVAL PERFL'COHOOCTANESDLPONYL FLUORIDE (POSF; T-7L4) TOXICITY STUDYBY INHALATION ADMINISTRATIONTO CDRATS FOR 13 WEEKS FOLLOWED BYA 4-WEEK RECOVERY PERIOD THStJnu.edtKyioDagnidryaenc, tBLo.riSf.ee S(Hciea&nc.)c.a Ltd. Dec Tbe sigaattae of tfao Study Director confirm* this protocol as the wotting dnrwm--tt fcr the stody. WimtingAwi T.Wa T 3SInMptwocBsuri,Bafacff ' PathHausentsiniggndobnotLhicfatpidtaaotfath.ispage, retainonefaryearrecords andreturnone to theStudyDirector Ftaai Protocol Exygen Research Page 104 of 217 : 621 MIN 313/023622 Exygen Study No.: 023-076 Study Number :MIN/313 Huntingdon ' Life Sciences FERFLOOROOCTANESULFONYL FLUORIDE (POSP; T-7A6L4) Toxicrrystudybt INHALATIONADMINISTRATION TO CDRATS FOR 13WEEKS FOLLOWED BY A 4-WERK RECOVERY PERIOD Enquiry Number: 23923B Numberatpages for internal distribution; 22 Dait-ols c e u i 6 ' 2 a y , HfCPfEnacsn2rtd8bf1n4rriigHd^drgtSorenotakiir CflUtTO ADpcccodores tobopezfcsmedattbeabove sitoanless otherwisedetdiedbelow. l ocdk a tf^ id flc ta d Blood rf* mitry RCCoeaunmttibnrere,ildEygydeeos,hnSierueaf,tfPoHElkn2.n8IiPi4c2Hg3dS7c,PabXRn:efmsaeraalyrocbgheisptiecrsf!orremnaetednrna*ti.iEwygetRHe^search Histology : RCCneannmttmrberr,iidEygydeeos,hnSierueaf,tfPoHElak2n,8tImP4g2Er3iSr7w,PbiXRutefmsoeeaalyrocbgheisCptieecnraftlorerre,maseodna*t.Eye Research Smchruymtai,vu-writnve,4E7vrverf>avri?ttef^st : Sponsor. Exygen Research 622 : Page 105 of 217 MIN 313/023622 Exygen Study No.: 023-076 St*dyNumber :MIN/313 Q0N1BQ8 1. INTRODUCTION 2222..321... STUDYDISdcSaharCeig&dH5aEyotecDccfoiUitfriLtmmrErejtAfapteNake<DainFTSoToRpUaCTURE ' 3. TBSTSUBSTANCEANDATMOSPHEREGENERATION 444.21. ANIMALMANAGEMENT AAsians-hcia^ dietmd inieriutpv4piyTUv 3. INHALATIONPROCEDURES 555.312.. TWtf | m>wAir 65666666666....-..3314246275........ SERIALOBSERVATIONS BHMFBUcmolorodoarcoayadudUaloByrctortyheitojaibgatyim, pyeripheral Mood Urtao SEMX-try eksxsi.analyse 777777...32422.. NECRMMFOFOrPnberoctStthrtmMYoiooedAccBHooNvwpfsDIid^rcUjsHPtotIatStphTloOeloiLgOyGY 77..67.. PSCatNoAbcsty*ba$fixcellprodfentjeo(Tabi1) 88-.1. PATHLOiLgOhtGmYtercaccpy 8991..031... DARTEAPPSOThtaRoRstToEarItAmiNmTiGgiMogEurNlaypTrhisy(Optional) I11LL1I2. QUALITYASSURANCEAND ARCHIVING PROCEDURES Archivine H u n tin g d o n Life Sciences Page 3 4455 67 77t 11110000 111122 1111111133536462 2111111iIS077778t7 222000 21 2211 22 . 2222 Fbml Protocol Exygen Research Page 106 of217 : 623 : MIN 313/023622 Exygen Study No.: 023-076 StudyNumber :MDV313 Huntingdon Life Sciences L INTRODUCTION Managementofstudy r Stady Director . * : TJ. Kenny MoritcriagToodcologfat : D.W. Coomb IDtfalgktcmhcetoctecvm,etrphboeyrsatcrhiyeeanMbtisfoeincnicrteoesropinfogtnhTseiboSqitlcuiiodirlysogwisiltl;be tshtoelfiirdstbienRstiafnincec.d to tbe followingpezxnin : R.Davies Ohjactha AefsfevcatasrwuirDntboefesvyagleunataecdtdourreinpgcatr4stwiaeleikaraec1o3vwerrydpreirniohda.atfxr study in CDRett. Recoveryfrom ay TbestadywiDbe performedincompliance withthofollowingregulations orguidelines: Guoiciina far the Testing ofCheanoU-413. GoodLaboratory Practice Tsehtefosetatbdyinw: ill be cendpcttd in compliance.with priacipks of GoodLaboratory Practice Standards as The UKGoodLabcntoy PracticeRegulation 1999 (StmnoryIsstnrmec! No 3106). OECDPtmapka ofGood LaboratoryPractice(ts revised in 1997),ENV/MCCHEM(9$)17. BCCcmmissioQ DimtLro 199*1l/ECof8 M idi 1999(OfficalJam al No L 77/8). Animals(.ScientificPrecedares) Act1936compliance TcppateardrhcoommfictvmieeninsdivpiqisuoBaotrneiefenserss,eaoMsdrmufx^bflatwfyhyterhetriheTibnTUeietgsenUr,stimdfatKaeci?sadHtopdrmfeKnepsmctisargnoe<bcgjfrdaOealoc,rafetmfsftlilhtici\cAeende,gnnfrticbremr"eagasaruwmlflsoantar(enSysspdcaeiopleclrfniosftdcviinfeiniaicddcrmaerptPrserigfrofdioogtchcrrmepaetdchmdfoueyimcrvdrecreiesddrss)tanirorgAtenflngicwfattitewiiseno1hkln9io.8v<o6ipfnpnegw(rtsihyatfafengtbfiipAmlwiscafhtnlt)mshh.weejwnThptithsrcatehwocAtmh!incecaeratyel, TtuhniedisHersotSaucdshiynewdguiallnledc1oCtmoaprtohlyeowAficAtht,naiimsBsaualepsdpuluiscneaddbeliressaeSoccctiiteoionncti2sf1iocofPtfhrtoheceAeAdcuctrta.ensdrtahdetlbaosoHrioammadnCoKdHesiiongf PerfaActnicime faolsr Research Page 107 of217 MIN 313/023622 Exygen Study No.: 023-076 StodyNumber :MDW13 H u n tin g d o n Life Sciences tTuhnhedisHeriotSoocdahymewgduiallnledc1oCtmoaprtehlyeowAfiActhtn,aitmallmaalepsdptlroiacedadbelirensyeS*c`c7titiWeonnTtiT2f1iocofPtfhrtoheceAeAdcucrmtesladnidetheyHvuymHanCe oKdillinogfPorfaActnicime faolsr The ganber of miraah uaed wjH be the camjmnm that h nxffia*mTwith integrity and regnlatoiyacceptability, coosideraiianhaving beengivm tothe wdfice of inifividaal animals in tmn* O f the errtrart rrfpfVK'flhjm tO b e T M Tp *^ OUt 031 anirrf l ; CdsDtaKavmai,lsabcciee.ptedbyrrgaiatoryagendo, background By Inhalation Group Compound Exposure level (ppm) 12 3 4 Control ------ POSF------ 0 30 100 300 2. STUDYSCHEDULI ANDSTRUCTURE r v 2d, Duration oftreatment Minimum period : 13wed followed by a 4-weekReocrwayperiod. TTaadnhhdiremiotuiatorglwnsh.afollunotwbansnteyrpvesarutiicoohdn*amdcdaoiytniaobindeaclerextdrterauatdemeeedwn,tuwpriyetf,raiodtdohceturemSapetnmotneesndotrh'ws icnaoncoaocenaatrmta, ratieomfeionarctoatrloplospruarrotveciovoaionnLyg AoclfBotsrseausatrtvtmoivrtihnnregfsMotararttihnoefstnRuedeccyrooavpnesiyrmyoapflsahnawsimiellatbnlseaantlraleolaac-taewtdeidHthtrosottuahrgethMoonuatitnhtheetiofridrnyse.tcdroapysyofpethrieodst;ucdeyss%atiwokn TOhfaenRyecovery paefribod miafynbcceceaxaatetynid/eifdV,?wiTiWth#rtphrmelSmpaonnsiroiHrsTcvolmnsaeiMnttt,nto establish the Recovery Tocbobebmsriecnprndvlgueahtcteitoo'ewondiesienawkntihylelbefbaifdenodarrieletcircoeoonprcadoolermdtsataputodoaymppeprgeotrpioroifdaetseithaiennrtdetrhvdeaulM*rina(mgSeocthtrieoRnenc6eo)c.vroeDrpysayptahpafesoerrionadencsyrotaphdsedieistaieowrniiaaQll Final Protocol Exygen Research Page 108 of 217 : 625 : MIN 313/023622 Exygen Study No.: 023-076 StudyNumber : MIN/313 H u n tin g d o n * LifeSdences 1 1 Scheduledtime plan Animals to arrive Treatment to commence Tenamal tof-faingica ffiatopathology tobe completed Draftreport to beissued . Docenber200l December 2001 Much20(22 June2002 July 2002 ( tmatnd) (estimated) 2*3. Identity of treatment group (to be elected from 118animahordered) Group 1 2 3 i4 Treatment Control Tatmbtance Test substance Tet; substance Expo(psujrne)k*el 0 30 100 300 5 5 5 3 5 3MaMleaiaXFeomdrale NMRuaamekobreerroyFfpeamhnaiamsleeals MSaslieeffiteFsetumdayle s5 s 5 5 5 5 5 0 0 0 0 s5s5 5 3 Main study Group Caga numbers Animal numbers Mak Female Mak Female r i 3 1-5 21-25 2 2 6 6-10 26-30 3 3 7 11-13 31-35 4. 4 3 16-20 36-40 K atm ar phase CageBombers Animal ambers Mak Fcauk Mak Fesnak 9 13 41-45 61-65 10 14 46-50 66-70 11 15 51-55 71-75 12 16 56-60 76-80 Exygen Research Final Protor*! 626 : Page 109 of 217 MIN 313/023622 Exygen Study No.: 023-076 Study Number : MDW13 H u n tin g d o n Life Sciences Groarps 1 2 3 4 Satellite shady Cage numbers Anfana! members Malo Female Male Female 17 19 81-85 91-95 - - - -- - - 18 20 86-90 96-100 3. TEST SUBSTANCEANDATMOSPHERE GENERATION albtriehaenyggedoiatairrthtldhdaaeeeidtniryCSogfcpoofkoloarnanntHoztsdowotmhrlrneonegtiofSonorSpgiuaHogtbrcikomscotcatLsrinnciscfgoehe*doahSolcHslccszpmikm7rLnoiadincvrfsgdceidacs*eShtioHcipitecnmononcemHteniapestga.loydlwftohwsTictuRhihbtlhceistfhgtdaefSunlspShcaotcetHftnisiccsedbcroal.sycirlnts1rhsgo9haa9wand9lid,l,thriSaatailailtltf,oleaaitoenyccfafoaaaoomatnrrdmpWyaxlahtygeitr.oiafowrokMnioeHtahftrvcvu.aaniAlldltceidctrtuc1ark9tro7ien4nigt,t SbbSoencicrhehenacnicenedifsvloe,erdodmtibhasyetirtoShdnaenfifelnmatybtwfoMaornaafwtnooarritgywse.wgmcAfeoantntptthaSflieernoridvpniifgscrctetwrhseolatiHetoqBanuaatiofavaftaliSlgneaignOfdtePoitynplfilMbfinefaneSnaaSdtgiccOeiaDmetnmetc*naetytseSbbfeeterftvaooicrbcceecedspaitaaeactbt.eBelesad.taanttfganhsgahdfexOenBtc*tLocaiafsnet lsaaffoemtywprreiccnaurteicoenisvetadkwasililnbtehneswedortkopsleaccteh.e HuntingdonLife Sciences Hazard Class,which*dctmtuncs HuntingdonLife WjwMn w Final Protocol Exygen Research : 627 : Page 110 of217 MIN 313/023622 Exygen Study No.: 023-076 StadyNa*ber :MD313 H u n tin g d o n Life Sciences 34, Tcatnixam SRptnoynaagonrr'saraytteiTnptiafyicijticp Sponaoc^ifgpoiaTnKtfcs <Ado:reoammctnebedisebtnoytdtthninentpShpeeortaotaorodery.odAraintnayaaanraedcfhrmidgeoefacradiatookdrm,icnwotihllaol bsfieontahlerrewpiosret. Ddeorcivmataieonnt.xtjcc ofmethod*ofsynthasi, fabricationar S ta b ility date. Cfffrf^-gtw rtfnpfyrif 4. ANIMALMANAGEMENT 4X Mhnab-snppfr, acchaatbatin ad aBocatioa 4 JU . Aaimau Strain Aga ordered Weightrange ordered Supplier Rat. CrtCDBR. 422dtys. To bewhhinan 11g rango foreach sex. Orarie* River(UK)Limited. Duration At least7day*beforecocmeaccraeat oftrwamot. Referto Scctioc42. 403. Aflocatioo to m iMBctgroppa Allocation Motbod Cage dbtnbctk OaanivaL mr ATiaodftroevqriduthanKea]fstocaatragtreovibfraoetcxtrpentoyca.ontrEaelasicenhafclbuhaetgntrecoreyespwbwe2t1iwlbloebeoabokgoeropsoetpodshn.iaan Final Protocol Exygen Research Page 111 of 217 : 628 : M IN 313/023622 Exygen Study No.*. 023-076 StadyNomber :MIN/313 Huntingdon Life Sciences Method Cage label* Unique foe-- Tail tattoo. withinitody. TTr^igraTy UVntrfyrng m r w pirf p1e0rsiopda.re win be orderedto replace any zadhrUadsrejectedduring the acclimatisation Replacement beforetreatment : ABIBob-dnhyoewsrjateahiB.ghtftcrsangeextranet. Rwp]t* Aii?iigttrjfnifnf ; NOOC 4 1 Animals- bearing diet sad watar snppiy Kodentfodhty B*wlr^frtTy~mm4nw4wmtTyr<f>tiw^n|biologv^l tnH Air supply TRedmaprierreatbusrme aSty Monitored cantznoooaiy or daily. T.lgbtmg ; Aiann systems t Filtered, notrncireulatod. Mlinttoaedwithintherange of 19-23*C. Maintainedwithinthe range of40-70%. <bree <Wiun^int jg study 12hours light: 12boors dark. ArtirntrA m trrrrtTlrfir^ ftfKrwwyrw4 m y q m e tmiiBty Electricity supply : Pobbcsupplywithntomatk: stand-byge&crstarx. final Protocol Exygen Research : 629 : Page 112 of217 MIN 313/023622 Exygen Study No.: 023-076 Study Number : MIN/313 H u n tin g d o n Life Sciences Animals per cage Ctge tutorial C^igr flooring five ofthe scoe sex,mies? rodaccd by mortality or isolaricc. Polycarbcostoor starale**steeL Stsmxas c t grid. 1ianp5tperrcovapaglrseia*etaewciihnQtwebreveeasklu*;s.cpaPegnreedsce,idcsaeagbdeoe^tvareailaysbsos,offeocboaegdaihenogppawppieelrrls.bteTaihdaedwluaadttteeerdrbvienreilttllhebesef-cwinhaEalnbrgeeepcdohraatt.nagpepdroatpriate A23. Diet sad m tor supply Copka ofallcertificate* ofanalysisarestaredin thearchival. Die*supply Diet rame RpraetsseondtiMn tohuissedNieot.. 1MaintenanceDiet. Fiah n o i is not Diet type Availability Pefleted diet . Nha<enmwateo*ljofgcyttdor(eMxcoeopdtcObTarrdadstgibyl).forblood umpire* for Certificatioo BfmSboeueirfcpfoovrproarleebriieaidoorneu'lsyolsigavbnnieascarttayrclyilhretciioaooccfGnahdltiaibclmeegttrt^ietinis^bfaYirncoentalsffetVdea`sisreieaftdlrio*afTasa|dcrnruausltyei.sneisdebdyatnbdeaspupprpolvieedr This diete suo added antibiotic oc orprophylactic agent. SojipiT Regulatory agency Avaabiry Certificatioo Mff rirrnlrmymy^fry UJL Depaxtnrru cftheEanramnccL Naiweitricted via polyetfayicior polycarbonate boette wife Certificatesofanalysts are:routinelyreceivedfrom thesuppBer. Anal Protocol Exygen Research Page 113 of 217 : 630 : MIN 313/023622 Exygen Study No.: 023-076 StodyNtmber :MIN/313 H u n tin g d o n Life Sciences cmiIntoaiefsyitstabhhmeemtpSaepomrolftnoa*srsmloeikrde'edsiloyrieftssroapempqcrnueetejisobmtdieliidtncteaybiytihddtieheaeotdsovS.sipcsooenmBsoonrnc.tfmPfePgedOosStna,dlaiyfm.eeAStancbiaeolynliscteeessofofcfPraOsnoSydFsi,pcmeccamiyfiabcgeaparnesssent 5. INHALATIONPROCEDURES 5J.. Pre-etndy dxarsctnisatiCB BexepfoorttedopcavmapDoCoDrcceoaoccBetnotrfattrieoactmi wenitthtohuetsayntitmnanlswiinnobredechrator:acterisedt thetarget loved*m the chamber. 5JL Test atmoapbcre fcaeratfen : AHglam vaporiser. I(btoooteagthswobfccktsilace (bIqloitkjl*) wpiinllebde)meterednto the vaporiser TGAbpeovTaOpTTo-octh/aimlrbmero.imc prodocedpatte directly intothe t TiarnrMidnngffdrrfnmifgfi<wysyfwfryriitn-lgnfgter-srmTer)ifnfretaeiadopfrpmthrtoerp^Trcri*oaTtnetrvocl^lendgbay. ^th4e)1hm 53. Tast atmnaphes* adinlniatralfoB Rents Exposure system : Inhalation-cocct oniy exposure. AahcleaDlnvoOtiyrnaecglxo2apme0orpseoruxsirposeiolndisgnuiarlaeembtpwabositert^thssnu,aiottaaddnadgaveytanorCtpiiaoaabelcalaetcirtrsaioaiesnmdleorbton.ecrlcacrpifotsaeemactftiaiaorgnnnsaneach Final Protocol Exygen Research Page 114 of 217 : 631 : MIN 313/023622 Exygen Study No.: 023-076 Study Number :M3N/313 H u n tin g d o n Life Sciences Controls (Grucp 1) Sham doting Daratkmofdaily exposure Airflow pemiclcnexhrkvoepastseodhemoaleptessd*btauec(erprw1rhraemrsat,iot-etmhiCthnrrCThbuobeefhedudcxretigmsfpnwhftragslekbittslfwtoealetrrgriobbrwUanmeepi.inbiranntioAedbngbaveghnseai.eitesemuulnedcsbddaeeiw)*lnd.seoewfrxTdfoepibdstHrohteyersebaapuaitceorcnnehscaibinpotgettgior*raodorctinituanhnsbp(etgnole.deotstvDotthoweiuaelnci)rteihuhonxaspgftmehtobewbscieurirrcnembe Ecaabcihneetxtpooasuvroeidsypsotsesmibwleicnrboeoh<ooursieadininmatrsioepnabrcattwe etmxtragcrtoups. Azrcoly. Trtbchaeefftiaarnisnitmt.tenaslotsrsomunbalsslttyaondocyveewerixanp5obdseuaaryecp.deurniottdeidmtmo etbdeiamteelythpordecoefding : 6fTobbrooTroahndu.ar5ysddsaoayyt,hdaemtaicdshatywbeaefeitnke.trhtFbeierfsifntianelxaplweoexsepukroensuewcreirl.olpbseycabsacheduled 30Vminute. FSopccruieorr-.teansforaedinfraredspectroscopy tPlik). Suppliedby 5wAX Twi atmosphere sampling Sampling : ScahmampibeesratfbtrboeotgeasttbaetmFoUspRheinrestwrufmflebnet.drawn fromeach test Concentration : FteTmIRpo(raatlfvreaqriuaetinotni)n.tervalsto demoesaate adeqeme eruttai of AtTcSbcehnencidFsStiiomplttincaRe,ontAhssnooemurdrlaooaynlsysoodegblztyeli,enwmwcsfiwoicltldDhibilfosleialbeofldoajaylvabltoaoltelwhiddmeeaAdedtetnibhdnaeolsprydtersoiHdtnlsoocu.uginpytoliewnf,gtibadfiieotbbnHeoleesiagufdehpposSlpfideeecdaifbciyecs. F ini Protocol Exygen Research Page 115 of 217 : 632 : MIN 313/023622 Exygen Study No.: 023-076 Stady Number -.MD313 H u n tin g d o n Life Sciences ^ 6. SERIAL OBSERVATIONS FmToheeertephrtehoaacnruier2itadilma4yesws.oeTdfhaaefrloeefaxtfrsteeaarstnwmaeeteincktapn*ruewmcibiltleebrdseaqiyunon&tuexmdiebmderamsy, btwhehevefrsoeiaeqldoroebtpyeodnr,ot.amnoyrebethvaanri1edwbeeyke.ot 6J. fltnkal obeerrstk* Axmmls 1 theircages Drreeceaorpireaddteidoofoiftraocmtinmoerminal fcapcctedat kaetwico dailyforevidence of reaction ft- n r iT l-hrw hh NDDaatattneardaeonadcnatdinmadeeproerxofigaryrrea.srs;.ofthe observedcoodicioc. Qpoeeach wed:forall animala Ifnollaodwriifnrigocf.redqeoteaoilceyd: observances will be made daily, oc days of exponte emtfinj to the 423L... PDAArueasr-lieeanaxtgcepheaox*wapmporemaouooarbi&eaies[lOrcrveibatceuddercnrove.wtdtiotoodcmsegvedraeyly. restricted tiringtabe restrain!) Theabove schedulewillbe amended, anecessary,in the lightof signsobaerved. A2. Mortality Debihttted nrrcnah Ocsbnsreorhviehdscmarefully,may be isolated to prevent P rp m jfirr ^ar r if w ; A tw rw 1 m ry H TW 4 rtn Im niiM i flrrrra H nr it cocndered m tm tm u . WtbshepraerapmraectteircsasbpleecbilfoieoddmsamSepclteisonwsil6l5beatnadke6n.6b.eforedeath and samples willbe analysed for Animals fo ignroeuz ntidcsn d z oordcmdetamd,mkaisllced : pAAprnreonismveericadvrleoesdpdfsofiynourniastdMrpoeefcirrptfisogoiredrmrpeaeotthdoeer.'(naosporpmoronaxlaiwsmpoaortkesldsyiaby4lew*.Qill be VU.I JfEEal rTOvOCOt Exygen Research Page 116 of 217 : 633 : MIN 313/023622 Exygen Study No.: 023-076 Study Number : MHV313 H u n tin g d o n Life Sciences C ' 63. BadymifU Bodyweigfar recording : BWATWteenyheerkcwtt-rte1ofep.tXksM.yC.BBBHtCOBBB dispixfyrirnrgjnmrn-tbem^etiojh. mTgheirstmeydVaitwapwwiDft rbmetrseitatfi>n>eidrfmfhwAme rarrecfhnivrwe*n./if ri--cy yl^jon r tf TrtrrefTy (A Food ccesnmptlcn WWeeeekkl-y1. . Food sopprd Food jpJed Food remaning At intBonli each week. Recocdedat cage cleaning. Recorded at endof stndyweek. iS . HaanatoJogy,peripheral Wood Week 14 (Day91) 18 (Day 121) Animals Main groopanimals Recovery grocp animals Condition AnaesthjeJttiCc : Fy(doaolsclroydwelicnJeOgdBeoCnvStestrf)an.citgSUhaLtmdpelpebsvcaotlilaenctoedffgoooddearnLdigfwcttaer TtrtfhirT^ Retro-octttal ainas. mQn/DteJ/OOS^mtoLL ADsanfcs wiDbe examinedfer thefollowingciugictemticx: 1) UsingEDTAas anticoagulantHarmatnrrit ETRDroeiyfttfiatechlrurleoleoncuctyciyatoetlcekcysooteccacotoccyatet cFohctnatl P.rot.ocol Exygen Research Page 117 of 217 : 634 : MIN 313/023622 Exygen Study No.: 023-076 Study Nmaber :MIN/313 H u n tin g d o n Life Sciences / APMMMtbaeeeoraaootnnrntcccaaeeeacllltolliixuhhvianaoceotstmaqcnoofeggtJbioocbbbiinlaocoadafciclmmraflCE 2) Usingcitrate as antk3*$nlzntPArcottihvractmedbpmarttuinlic.thromboplagip tin (L6. Blood daacaittrj Week 14 (Dey 91) 18(Day 121} Animals MMmgmif minali Recovery poopjamaali rnrrfjtxwy : F(gcoeonllteodrwaelcinnegdeonvttehin).rngSfaas,tmtfperlisvacEotHicocicoodf fuonoddecroligghwtater Anaotbctic ; Iscfhgaac. Sample ske ; Retro-crtexl ass. Ant^*^g"?TT*^*TT,r1^ vwfarrw>. Litfaimabepexin/Q.7mL All tempi**willbe far die fcUowing characteristics: Using Hthwnn ilI a s T tfifn a g n lT tf -- AAADisapbxorSrnttmaeleipnahnmoasoinp-tohr-aattnraassnfecsfrearsacs(eO(TGTO)T) GBTTCCUrozhSnitetogiaaaomlllteyiwp&scbtreeimorsriteode-.ltieon~statoltal A l>mrwn/g}fhnTw nfin fbupfeotu P M P w tnrol Exygen Research Page 118 of 217 : 635 : MIN 313/023622 Exygen Study No.: 023-076 Stodytomber :MBOTI3 H u n tin g d o n ' Life Sciences LBSCoVrDerNbaHitti+noeldPeebhnoyzedyprbmoogeiedsnnaassee 6.7. Crinalysia Week 4(Dr2S) 14 (Day 91) 18 (Day 121) r/nHmnni fttrflifri gmnp imrrTMTt Recovery gtmjpt tualt ; Ovrmigl*fartvffvMmljnwfSrdlrmraggnfh and wster deprivation-. aarnpt* will be etommedicr the fioowingcharacteristic*: AVPPSHprpooeplcuteoimafnriecangcreavity ~BSKRolero>dtroojmandVmepTing(mbenetpsigments) Ibocfcpocr, obtainedfora crrtrifhgitioa, wigbe esamnaerincrotcopcallyby LM for. LESCCprcaryoyasttcsnhstosarcaolystcotyextaeoa sad precursors Q t frrrr tW m a t iM -T y ravruM ACCranysdstsbtay-lsSEMfor Methodreference*will be detailedin thefinalreport. Final Protocol Exygen Research Page 119 of 217 : 636 : MIN 313/023622 Exygen Study No.: 023-076 Stud? Number : MIX'S13 H u n tin g d o n * Ufe Sciences iS . SRaemcopvteearyo(fDWayoo1d21(f)bmc sdcrSatmfe)t,ikuerin(Dea(1y 7m92))asnaidmlailvse. r win be 1necropsy fromMain (Dy 91), BstoalmofoopedleSwpafoflnllosbowere.rdemtoodvoedt iit!roooocmrotpeaaypbsyatcaarecEtaacd/atfoarptisceppmunucrturireciantmo ftarobzeesn(porpiotroto4 dmesl)p,attchhe Uy2f8rt/yi2nr9e<fcir,oonlmlpe?cS^traejtdretnBovioteefranwnigmimhatatfylsrrokimtillpMeadxaaoiznqnesDtocadzysR2a9esc.soavye.ryOgvreorunpigshatt DaTyTt9p1wawnidn1b2e1 wcoillllebcetefdroDzeany The frozensamples will be despatched tothe Sponsor'sPrincipal Investigatorforanalysis. Principal Investigator Address : LisaQcnea : 39SM3t.5EPBnavuoilsr.hoMcAmmvnennretuaaseloBLtaau,bi5lod5ai1nt3og3t2y-3-33E3-109 TelNo. j (6(65511))7777S^-66011786((0F)) AAtwhfnleflylSxbfapaeworrntdehstaoeatrria.nprerAeodlcecbevoysapsAnyintcogtoSfatptnbbodeenaSsanopnaroa.llynysssioserso'sofrftehtpeeossrtatsmwupbillsletbmemeiendcaldnnaddtaemdceoatlsalabatonioliantdewdseiinnlldpbulemasthmtoeart,ehuserpisontnuesdaiybnirdleiptEyovroet.rf &9. Urine SEMX-raydesaeat analydi KWIhnaodevMieevrkibndeg1ue3ranolaoaunlmrdlionRfwereoecsmdaomfvreeprelyeasgcwrcoeiuslspl nbtaoienmfic*omokoaldlleAsacidntreudidnrwignwaWgittehterbi.enekl2a4sbtoowfofreoseekfosoltlufodRwyei.cnMogva`elirinyg.hgtpTs^bo'pene'!ei*n>mthoienmAicTMmwnnftfgll Ithfefntetsbbevsoaimdpulreinweililsbneortemobotvaiendefdrofrmomtheauprrienpaarryabtiloanddoefr,foaetnaencimroaplssyw. ithin foe time allowed, TIonbbvteeaisuntriignfaoeteosrasametdptibimeeseUwnnti,ilvlaebnresdiitnyfoxoenfcPpdlriyeamptraioryuetadhssfSaoEyreMadnafsolytrusbpissHbdyueaSsipnEagMtachXne-adrcarytooccllcRemcatyerondtMeid,oecnnitmeti,tfriPicfartiitngiorcrnifp.taol iaunsw uu* 1--^ -A .1 Exygen Research Page 120 of 217 : 637 : MIN 313/023622 Exygen Study No.: 023-076 StudyNumber :MDW33 H u n tin g d o n Life Sciences NECROPSY AND HISTOLOGY 7JL Method ofkill Method intr^peritncicil odiumpentobetfritooc. Followedby 5t*^jurrir* To iQowatiictory inffir-grao? snpuiacso. MUA-- tna-cxoe-co-psi-c*xnn'oiogy (TaUel) Complete Checks Photography Spedai leqm rannii 73. Organ weights CTabiel) Pea coBectioo Data presentanoti 7A. P late (Table 1) Standard Others Ail animals. Retained tim e ii dUinsaoseotaklnceosfathipeencetecdrotpresytesaaepneMrvtiabottccodrfSintoddinygDs;aagtctthoec. Rapeptaroinprlyiamtep)h. nodesadjacentto messe*(where FwPoseteibgbohiileoadQtetronPgdneoxttrhxgefaarnztBas,kelsaeTf*tibaontihede.rriwgihsteosrpgeacnisfiwedinocbethe Okfrfgleadnowrediygihntgsaprreemnoatturroeulyti.nelyrecordod for anhsals AAbbaaohtertde.fortsnniaal bodywdgfa. 7kac Formalin. ETUyrrriefnslaf:ryIhnbwDlaiapdvdwiedrist-,honf'tasyfth'rnwd. nri lightly <4iti-Tvf the orixtry bladder withfizadre. Final Protocol Exygen Research Page 121 of 217 : 638 : MIN 313/023622 Exygen Study No.: 023-076 Study Number :MBV313 H u n tin g d o n ' Life Sciences 7i AtfhroleztSaeprnnobmnysaoilrml.rinTUwhoreifasMacmainipnlH,esRqnweidiclolxbmveerortagskenendnaSfnbadBtasotlowlirtmesdagnionirmgaaadlneswespaemfarpereilenzsegoapfteol-i7ev0te*drCawepiarlil.obretoredmeospvaetdchtatdo 1JL Histology (Table 1and Section 8.1) Procr.w n; AAUSlaKinEgflrildlihcMntsrntnaoaemrXaiiyymnck-ponborapedcltkayylaidkm.cmcdicfD:eipcGryMri:fotOacspcnedarxdcopodtabciyaoeelinaltntloonagpapdbrplotecy4tlciai.pfnuHera..e:ylsHBe,ainicduttaistyfihoni.eorgdoLostingzhhe.ethr haalflffofeix 4-5jnn sectioca sarinedwife haezaatoxylinand eoaizi. Sappoepctiofircicinbocdeideesr.to(yunrhatlvdy a *tbcpresence of 7.7. PO^StaiBbtig for cdl pro&fermdoo(Table 1) Sanecdti4onfeeoPfCuNri^niamrycmblpadcdstegriwriinllgbine otarkdeenrtforoamsseMssatihneadnedgSreaeteollficteelglrporuopliafenriamtiaoln*pinroGtersotc.p 1 A total c f3,000 cells wiEbe counted from 3 separate ancrincs (1.000 nrotbcrial/ssctxx) cells fr> rrrAtr t n t\*!r\iniwi f l y f r f l pTTdifa r a f to n tiy f e r . PseocstiitoivnesaPnCdNcAonint:tmS-ipnhgasceepllosswitiivnebceellsa,iidfperdacatincdabthlee. exammiog pathologist will assess the Ienraaifdrcpffiotiofocr. st&ecCtiioinnTHs cwfmdantaoidwrmcogijnjtftrtfoamodeoalocghya.nimalwillbe taken tad stained to act as positive Final Protocol Exygen Research : 639 : Page 122 of217 MIN 313/023622 Exygen Study No.: 023-076 Stadj Number :MW/313 TABLE praccte T J a m a o d n c ia a c to b ta M fa id A h u m lk to Adnsab A a a -fto n d c Br*n*dr*txr. Coke TVxVm n H u n tin g d o n * Life Sciences N arcw r Hi * - B k to k c r F a tta L% k:^cr]copy - A c a c ie bocks PCNA Fesaor(te e *to ta i acetico tiaouxh k*=0 H arfebanftadi H B ad a it __ (te la d la * aortcafar ta d nmcscalM ra*icoa) Ih n J iE i-- LtrvDltiMax L B jB flla to il twolia r (eakto fin s a ll sa^or toba) Loop (M etto*te a m *jar k*xs iadod&g L n lA - a d a k _ * -m to a tto n a e . M o n to - .. H ai to ttta n (3 M ) -_ ) * O p te -to '* '* 7 ____ . _______ 5-- I " " S to in i e u - a n h .................. _ r-rta* (. ___ ...ta .p f rr %- oarfkal, iarahar sod thoracic lo to ! *< Sam e " r a ir li T e n a ffA S ia ta to *) T im U lM lB W V !. Too T V naC iocladtofhttond ooh t * ^ * - * * _____________________________ Utero a riti: carrix ' V tffaa a) lSOaaeteptraiao^nwaMd>o^#IigfcKrt'tiHLawiclfytBp.ioppMKtomct^omMdlMroiliMrsftafiicMdfoaaailnraiWdiariaMdraygtox.aoopkjPy, Uf BnBnU?rTiTT.XnOrCnQnIt i: Exygen Research Page 123 of 217 : 640 : MIN 313/023622 Exygen Study No.: 023-076 Study Number :MIN/323 H u n tin g d o n Life Sciences & PATHOLOGY 8X Lightmfcroci yy Cte*7 ftananredcah. Terminal ** PccrRsvictr Item s All from *Hgxocpt. AHspecifiedmTable 1. AinlGlMroaunpsa2ndanRde4c.overyanimals AHspecified mTable 1. : - CmcriOOt by 1 g v i C T i n t o rrffmvitinriny 83. Sxtenskm cf Initial examination eAdviteatlahureactheiviaeadsc.itviiornia.cilfttehcecpsia*t.hoDloegtaiislt*,fcafrtthhce*reptroocchenssiginage*tawdilsltbaoindinogcatencxf.cTntitqtine*ardorseytabineeHdieind to lSip^ehntsmeri,carsofsoclolopwyso;f tbe specified tissue* may be extracted, fcrowing consultation with tbo tiossmoeaslclmaanaimdearlesdotof MexahiinbiatnadreRacetcioocvietroytrGeraotmupesnt2intaGdro3upti4lt.ed at trerninal sacrifice: for Anysneb reqairrmrot -willbedocomeatndin an amendraent totheprotocol. Tpffrevsaxeysrdwigsp<*layinmgttwzyeitncrnhxwolity-^idsialcblQgeafy be farther examinedosinsadditional S3. Pbotaakrogrvphy (Optional) images : HSpftoanfcsaotrk,mtakoefnmbayjoarPleastihoonl*ogaifstte.rconrafarkm with tbe Repost : Pall pbotcmtcrographic reportifreqmtcd. r '- Exygen Research : 641 : Page 124 of 217 MIN 313/023622 Exygen Study No.: 023-076 H u n tin g d o n Life Sciences 9. DATATREATMENT 9JL Statistical analysis TM*tjpa Thefollowingdatatypes willbeanalyacdateach timopoint aeptrately:fMboooroogdodaydnwcwcorhieegeithmgashin.stusetr.siyibonoasgtn,hgdoaavhibenaarseoamolpnvaptetcroroalzponpgodpyaar.toodpjsurtisuatdteryd:spftooerrdityoadpnsen.riinoidlsb,odyweightwhere appropriate. fmfKngJfqf th. mrrwTvr n f Tim a N xv4fri a m r f TgrtHrm? inr^ findTlg Peaecrhctarteeagtoerdicgarlodoaptav,etrhseusptrioiepocrotniotrnool.famends willbe analysedwing Haber'sBead to t foe Fwbcoeoitlnrwlycteohaeenrnantmotbbnoeesccgcworsomhduepaprtaesar.,eodBcUcwarcirnaittagihca;rtttyefhaa.ettoedsoteawpaeoinllldpfeiornsottpob,settDahpCepoolripueotdcraottomiategeaoagfdtBbjnaersbttlamertotc*omagfteetaTaeti,inttyirledoapftetvodargiraonucpes 10. REPORTING Study progress : PpRrerrrooAiuogntdrienitscewsvswmeyroiiblnolaipbltaeajirnspwdrreoowpvforitidhrttetesedntwbrvuiylnpitfdbhr.aeeptesSiasetSnaoOdtnya.sfDttueidrrye1cwtoer.ek Draft final report : ForreviewbytbeSponsor. Authorised final report After approval fromtbe Sponsor. Routinelyreporta aresupplied ooA4paper. Tbefollowingnumbers ofreportsare Emptied. Typo of report PHnttag DAuatfhtorreispeodrtfinal DDoouubbllee--asiiddeedd Photographicreport(ifany) Single-aided Bo0r011Nraudmber o f cUonpbie2001osund Aadndyeandddoiatifornmeonrdcmorernescttoiotnhsetfoinaanl arenptboartr.is-ed foal reportwfflbe doqnnartErftt a formal flsmafaumoocatdehbKnifevadi%cnnaabaresciivemgaennan(cos.se,raanodelefrliwesomcsnsutgaoieooirdntmibynseg,ioafcrsiownaamadpilldmplirrtbeuoipoenponirtcirraatatttnfeoiros)otnfahmesnre,rdtefHhidwniqsaitslolslitarbutegbedpdeysoourasQtbriwcxjehLimcilivtlfoteebon.eSthtrAhisVdenairytfaitooyesbnrotjbiierslsceacetsaoqeoeotrftveo.aenstftoirterhnqeduarteiiasgrfteths.ptfoItoonnr Final Protocol Exygen Research Page 125 of 217 : 642 : MIN 313/023622 Exygen Study No.: 023-076 Study Number : M3N/313 H u n tin g d o n Life Sciences O U- QOALnYASSURANCEANDARCHIVINGPHOCEDURES 11JL Quality Aaacraace The fciicwing-wiHbe inspected or auditedin relation to this study. Protocol Audit Studybated inspection* Report Audi: : Authorisedprotocol anday amendments. ; Critical phases ofteas study win be w specvxL Tobfethdeadfrtirfetproeprtocrot tdosttbuedySpdoantasowr.ill bo endued after issae QcoAmpfiancdtiionng*o-feilalcXh arcetpioonrt,eedxtcoepthtefoSrtupdroycDesisrebcatosredsaidnsCpeocmtipoannsy, wMhainchagwemillenbteprreopmoprttleydotoc 1LZ. Archiving Atblelprarowpedratytao, fsatmhepSlepsoannsodr.specimens arising from tbe performance of dot Study will retrain /--> rTeStcyeipneteniscoenosfSasntaadmndpaarlcerdhaiOvndipnesgrpaemtcinaimgyPebnreowcdehdiisucprhoessa.ered uonfsuafittaebrleth, ebypereriaosdosn sotfatiendstaibsilRitytm, ftioorgldocnngLteifaes SSASsthipccgleilioneensnnofwsccttothheeTr**iee.mrwfii,ssnialdailmtilscsrpopealcopertscosiahnrlitsov.oeoedArtfofosftnprereeirlatckaaipoirmnetcrhetibtnortes&admrxa^onat,f\dtntfehTirav)eiale<SlljsypfrseaotuwhanbesnjoedfrTcarwntottatmTiolTlwatbibaierlleelcadbosaoentJneffraueobrtteeclaeqdiwnufaeeehndesidctbebhhdoyi,tnsbgHWaedautSuvgatiirrutciiednneegygdarrotiDwHononigirtfheaLLcttiitobffoneeer Ba rcfoflpiymogdfctbneLfiifnealSrceipenocrteisnwitisllarrecthaiivnetbinedQefuinailtietylyA. ssurance records relevant to this study and Exygen Research Huai Protocol : 643 : Page 126 of 217 M IN 313/023622 Exygen Study No.: 023-076 FAZ:CHt0rS22tt 0.8718 f. 1 ... P. 001 N ^ T F n N n TOT iw w Starni trrtub m fi f v rr ta lin in ' t** 1 h* FAX HEADER SHEET ' Huntingdon LWicfrUengSfocr aibetantrcfimetsrt TOTALMiMBEXOfTAJ3MSINCLUDZMOTHUPAQ1: ? IfIMI m an tete k TOCii&flByyimlta n a u at Tit -M4(0) 1419to o * ATJJNIIOWOF: company: PAXNUMUDL M Bun D rSG m kiK m U iki txrtm X oank, 001 1417J1M f* * T iny Koany DA1& 1*9 September 1001 OO 1ITBTaIt)esboff(Fax *01 *91 733 IUi PwdtranfihilmiorirMetroNcat<aOmriUeteo)ilCfDOarjlateBfsoorriUdeV(/PnOkStTf;bOTe-w7eKdLb4y);>4T-WexeidetkjrrerctoovdeyrybpyortleadtU(SLtUatdi*y DterDrOnrtim. PprtoetuoecoOl nrdaeeotudcmhaendtl otgepryttocfpptrooltinocdonliaiymrotouddayxMarQJCtTelrZI.boeboveBody togetherwith >copy of Tny K ray LWorcoaal dfetcorlrtaSeBaefclaor -i*444411441100119911027J0I itetei.TB iatt.Bte.PIRteIN-Ta IDti KEB uSnEPe.19. U U PirT " ' Exygen Research : 644 : Page 127 of 217 MIN 313/023622 Exygen Study No.: 023-076 FAX:OlMOI922tl 2P.002 ; fSrnodntytN!rAainMtaTwb.aaat N m b tr I>M] DV313 Huntingdon Life Sciences FXMLTOROOCTANMCIJQNYI. FUIOXIDE CTOSi T-7M1/I) T tw ocrryrrD O Y tY inhalation admuottration to cd haw foe u w o n FOLLOWED EYA -W ine RECOVERYW OOD Ttttl s u W afpapt: 3 NnabtrfpafM&rlaimml4IstrfeflMi 3 StadFEfePMiar ; TJ.Krany.RfcCBoaaj. /--s Tg*itl*t*=imFi*tooni*ijoartcowii!4^dySdi4aatisayc>Dfon3atifcalttornrflaaudamaiotaroimifadddliiistthntrhaoopurgtiaihtnotiatdtutipgiTt uo. ronMf twMainniatbmneacd-oaAcnmnDaunacnz:*Wmpihirnnoottdo-cSotlnA. bnfaycr AMENDMENT AWROVAL ArtktSptHtg Aj*ao*uJby;________________________ Data: RECOVEB THE SEP.19. Hp=1 FRUIT TIfE SEP.19. lltM PK Exygen Research : 645 : Page 128 of 217 M IN 313/023622 Exygen Study No.: 023-076 ,t , FSCT \ l 220 2 tSmtltijmStmUkwi lri h i i i U f t i f ttKJDMU ' FAX:OIOM2I 518718 f- 3P. 003 ; i Huntingdon LifeSciences natsurokoocxA m tcoL raK Y L f l c o m b i tF o srj x -7*0 .4) TOXZCTTY STUDY BY INHALATION ADMINISTRATION TO CDKATSFORU WEDCS FOLLOWED BY A4-WEEKRECOVERYFKBIOD TBtwo SopmoofloerrMbaateatdoasfuedBttoNMipoZIinwtigiior T" b&t$ BSaemoopwle!iyo(fItbelyoo1d23()ibanrdnrSucatdOH.ttee(dDtaoyw29w)OImtoalct.ollected at imumsy from Main CD*y 92), BatoKloitaoopdkSwtp*oilUnlsoto*or%.mdtooocrleodt attxntmonrotpaotypbcaytekr*xdfaecd/attaofdmc ppwatma Awmt cinvtroi farlotos(purpiotroio4 dsaop])o,t6ab U"21r/i2a9*ofsoroooQmpabatbtaeotdoogttort*vfnuartifbn*aaltfyetscikrsialptMendtxafaantaDtiesdnyX2t9aa.sety,m. yOpwoconpisk sDon-ptt1savntdt!1t2o woofll!ktoota6deDaany Th. t e n t arapk* will w ta p ra h * tt* paugri ftfadp.1 tn M l,* fa, htmtfMm H h> M iw * **-- * st S fc M aiw Um s i iiiw M MWH IIHt Mi *** {( tf1l))7T71 1711( ((T0)) HECBVeB TDE SEP.19. lltlZ S fkp3 PRINT THE SEP.19. H = l* n Exygen Research : 646 : Page 129 of 217 MIN 313/023622 Exygen Study No.: 023-076 2l 02 F*X:0HMH22II Ka 8115 P- *p_004 Studywyi&amlA&jnn-ai.BonKv^w iffcf0rw"/t"i* Huntingdon Life Sciences Atridnddrrmg] S aM la tt i B rSJdm Jn.^ SASiAwSajsaS.aemJSdaySaoSkta laiNo. USA i m am a : aam m nw SroAneofapdjtoyVhUrswtfv.tSarrArtptawBolKlnfpfrsplaocbrwrSaa,cMdzaAaaaMttsaaciinoarfipeaadiyj*sw*bvovytdfdntif,thctt*itolSyStphp)o*ennssoszoefrur.i1lbyaroopaoairfpttkawcsitnsabudhedrtstaxpcxilocedosaHadsd&mmxxetawsbolbheadfteooernprcl%supstoh*oe,iiusbtriuiHadey^ g gTTVO THE SEP. 19. 1112PH ' PR2KT T IT E No SEP.1 9 . I i: i4 f ln Exygen Research : 647 : Page 130 of217 M IN 313/023622 Exygen Study No.: 023-076 F*1:0U0H22. IO' 8,!8 sp.005 . rSrteaBijcrNataAfMcvrtaHUNnbtr ::M1 DW13 Huntingdon Life Sciences raurLCOROOClAJOSBLFIW YI.TIUOSZSI (FOCTsT-7**lJ) KtSUMDiABY TOJOCrry STCDY*Y in h a la tio n abm o o stra tio n t o CD RATSTOt.1 WKXJC Total aaak e ofr a w 3 X ia h r a T ra torlaaraal (fttaikuOoj: 3 StaijDtraOOT : T.LKray,U c (Boat). (fiCOVEDTDE S p . 19. 1H12PH h*.l PRIMT TDE SEP.19. i l 'U fn Exygen Research : 648 : Page 131 of 217 M IN 313/023622 Exygen Study No.: 023-076 FAMHMI52H8 HO.87!! P. 6P. 008 li"" Huntingdon LifeSciences wtBK.TOfcoocrAi8mjwja.itt)OBnix(iow)T-7ij) . . rKU M W A K Jf WMUCtTVSTUDY * y balahon ASM ntunuTKm to CDKaTS FOR 1WEEK Tb* Spiot&fsrorrh iswdubrnainiitTcw4i*i Prinr;ip,tl 2bwtiptBr RECEIVED TITE SEP. 13. lU lW fl PRINT TUC SEP.19. h2kig:ie4lfln Exygen Research Page 132 of 217 : 649 : M IN 313/023622 Exygen Study No.: 023-076 i!-sE.sF-.]?-.i0P2i!?i!3A)(im L liPM J20 2E02 F U :tl4 IO m itl sa 8718 P. 7P.007 Si ncat edsjrKi uAbaourA n i a t N u l K r :MBuni :J Huntingdon Life Sciences Tog nbrtM m /m ntM bttlM iso ly w pSturmWiopsrlt*eoUid*ai*sfp&bal,toehotddoo(tb^t tMcSpmnaiarianOrw.^oWy crcwalatne(uapMtoT' o4pmcylbj.yrandU&*oc/ai amrt&ipiunpc*tamroa>dna)ddnmoimssbo Tbc flom am pi ili bsdeipscbad ts thSparar**PrfodpalIovaloor fixsudym. Mncipel taratipxor : LteChmcs. Adirasi -r '3SS9M3L551?E3BsmuMolr.ibMMroAaf1Ea.vaMUMaSBsoAtsit,IaBLuab&otatntofr2y-3E-09 TdNo. i3511*-7777WS410716f<t(Fm). PArdtaAcriewttJ im n in mr Td No. Putfrn. SUFAtSiAt1i6f8i0B1lln i mi 114231ton ; ISMailitt Mr*otpfjlOMybK--sfl.apTAOUt leMlnnpcii*ro.StdiAieums*aMoi.tffkTmnOnifilkluo.tUAo*oluKof^cA'moitoitp*aofvnmwiiilcrIuaIbdhcS*uim itxnoilddtaMjtBmUnafefnofUlliWuonnAidnu.pnlogtssoemAom.wMufxliufbay*- RECEIVED TUE SEP.19. 11:13*1 hpl PRINT TUE SEP.19. ll> l r l Exygen Research Page 133 of217 : 650 : M IN 313/023622 Exygen Study No.: 023-076 OCT.24.2003 8:44AH EDICAL220 2E 02 JO.2805 . 5 Stadyf iu ta r : MINOIS Huntingdon . Life Sciences PERLDOROOCTANESDIJFONYLFLDOKmeCTOSf;T-7L4) TOnCTTYSTDDVBY INHALATIONADMEOSTBATIONTO CDBATSJOB13WEEKS FOLLOWEDBYA4-WEEKBECOVESTPERIOD MiticaalariytkrintafeodologytateOi && T s a w ta te jx w te ta lite sn rij* SBaamepolmesya(tDbalyoo1d23()foradmScvtaolQl,itaic(rDifaiyrt2r9w)aQaataakc.oitoctedu Mcrapsytun Main(Day92X StfaoopoatdeSwpafalHalcatnac-.etdaetoaodvoadt aittraooaumotpnaaypbmy tsspnKdjaitacticppaanstteodrauinatso fErostwn(prpiotorto4 dkacI)plattctfae i3Xf8Ai/t2ifa9icfooromoUmpaiSoaattdreioBvoiatkabunninMaufiljsetIekdsOyMaudmoaaDuimdayS2aU9n.cyom. yO|rwota|tytt tDesmyp9i1nwdin3W21mSSIataaadfraDnaay Ita a m samptotwillta tapatetad to (ta Spose**Principal Isvasdpic*forasalysis. Priadprifcn*ad**cr : DrEiciatdCrime Add : S3SUx0tASay5tA4e1tw6RC8Ba0oa2ata-amupcrkcbDriva TParixNNoo.. 001824231*8033 0038142311380 RECETD T if t OCT.24. 9J48RN P*fo2 PRINT T ift OCT.24. 9:50RP1 Exygen Research : 651 : Page 134 of 217 M IN 313/023622 Exygen Study No.: 023-076 OCT. 24. 2003 8:44AM ' MEDICAL 220 2E 02 HO. 2609 f. 6 gSrtuodtaycNtjuAmrobaeardmmrt Number *:4MW313 Huntingdon . Life Sciences PESyLCOXOOClANKSULTONYLiLUOWDI (POSF;T-76*L4) TOXICnY STUDYBY INHALATIONADMDOSXRATIONTO CD RATS FOR 13 WEEKS 70UL0WKD BYA 4-WEEKRECOVERYPERIOD ToC*2*nzabrfp*ij*c 3 JfHDbc-ofpeg* tar lateral dfctribgtio*:;3 Stady Dtrd*j- : TJ. Ksray, B-3c(Horn.). Tcthfhiasessa*fimr*aenttretfat|nknTtiens*ft'afddtsyeldSeMttoeddifyB*Difahc*r*t*tobre+adsxUtbaeootsfftcrfwotidetetottthilnmoorpoikeibianwtsiaiepdsaftmlaonvcwo*fffytlhb--iesdtomcaivnahMnaDtPmCtrKtiotrlfpifaraiowatfolicmoflA.acaITqr AMENDMENT APPROVAL 7ra*paer ABCTW Jlur j- *** ^ S ^ m L bG sobs Dm " . RECEIVED TITC OCT.24. 9:48P*1 Page! PRINT THE OCT.24. 9:S0PN Exygen Research : 652 : Page 135 of 217 MIN 313/023622 Exygen Study No.: 023-076 OCt. 24. 2003 8:UAH KEDICAI 220 U 02 SD. 2609 P. 7 PricatdoycNaluAmsbsaeardmcal Nsabcr ::M4 N13 Huntingdon Life Sciences st -- hifffflttmhaifaKSf.prosnjrfOA SrnnhlwffllwtMhwdiDfKgPiBliffli'rinRiTwaiMltolBAt^Bi^mAlniagiTtni BM I eaAafqdtojbUnefmaStAphwoHecrfanfpolwbcro.eednAacmaaclloocrpfeaydtzeobondyfamttthheleoiSSjapiwbcmaaeomoafrt.b'bewr'nepa&ofprifewiaeitBtusbdbet4ffmnacetotcrcueidhdtaiaoscoaoawbiollfebaes4uk npptioemathriei,bm&eddiatyy received tite: oct.e4. 9:000 h | 3 PRIrfT TI: OCT.24. 91500(1 Exygen Research : 653 : Page 136 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exygen Research : 654 : Page 137 of 217 MIN 313/023622 Exygen Study No.: 023-076 PRODDTitoeOv,oiCaftOiOoLcncNuDrurEmeVnbceIeAr:.T,ION2 Pape 1 of 1 ExygenStudyNumber 023-076 ProtocolNumber MIN/313 Abowqufeafa0isdvn.ar9etopi8-tppp2apol.tiieieondTdtn,hcChreaoeolmpiwtbhoeerertavhtctseoiuorwd,rnvtesfecrt,aeutttrttevhthtseestnwehhxopaatwsrtrianaannnttIeeaotodlnybueazasegerpdacoison(idtnrsaratttettidlahsaitsedtieic0onta.n1ol9Qtoo8puf22rt90colC.iov9eare8rrA.5teo^lWmaiMbtuiheoDesnatn8btsaehtaecotiobssrtttaariectinialstelttldiooc.unatlAlroiopeuerot.flifinTeietrhtmteeenasytt Adeviationwas Issued. ACTKHg.IAKEN Tsahmepreleiss.no Recorded impact onstudy C since theVIamnapiayct-etoocf InnStTetrepsotyw^as not Deto: >17*03 detected inanyof the SgMun (sp<cri r-}^, 1. D^SVRBISSBQKRIKRSd" baia iDUatta-e<nL/L. - e i ExyganQAURavtaw /K.-' 7//.> ~ iB SiS6iii Exygen Research : 655 : Page 138 of 217 MIN 313/023622 Exygen Study No.: 023-076 APPENDIX B Analytical Method: Method of Analysis for the Determination PePon(efEtrPafxdleRyuregofcaelratunofLoloMuricvooteehratroRneh,oxeoeScasvdteuniarslNenfuosooonmun.ilc1aEfao)tAxnenMdca(iPtdU-eF0r((2OPiP3nFSF-e0)OH7aAS1n,))d,in Exygen Research : 656 : Page 139 of217 MIN 313/023622 Exygen Study No.: 023-076 TITffi PoMflaohcotodooefUAnnraaulylfsoeifuottr (dFiePODSa)aannndnPiteinoteodoecfaFfalofcinfoooncfOamraonicetAiddfdco(EalFeO(FAF)HinSR),ai Uver, Seriimmd Urine Rrnsicm 1 authors John Flaherty, Kates Ruhi, aod Emily Decker PATEISSUED November 20,2002 SPONSOR 3M M e3dMialCDeenpteari,tBmusimldiCnagr2p2a0n-t2eETOo2xicology SLPmUMN 55144-1000 PERFORMING LABORATORY 30E5xSyRgeesseRarecsheaDrcahve Stele College, PA 16S01 M B H M uaaira ExM-023-071 RevieioQ 1 TOTAL NUMBER OF PAGES 43 Exygen Research : 657 : Page 140 of217 MIN 313/023622 Exygen Study No.: 023-076 ExjrjcaMethodNo;ExM-Q2S-C7I Reruicn 1 MANAGEMENT APPROVAL ' /ioLETaaxihrbynaogreFfaoltaoRhreeyrsMteyaarnchager //A *!fZ f f*Z JS3oMphonMnLseo.drBiRcxeattpleDrnehesopenfaftratmtiveent Corporate Dale Toxicology ExyjcnBeseech Exygen Research : 658 : Pits 2of43 Page 141 of 217 MIN 313/023622 Exygen Study No.: 023-076 Eiyjco MethodNo: EiM-023~<771Revixkx: 1 TABLE OF oe N TEN TS ' m u s -----------------------------;---------------------------------------- :-- i TMAABNLAEGOEPMCEONNTTAEPNPTRSO--V--A--L--------------------------------------------------------------------------------------------------------------------------------------------- 32 LIST OF TABLES-------------------------------------------------- :------------------------------------- 4 LLISSTUOMF MFIAGRUYR_E_S_-_--_-_--_-_--_-_--_-_--:-_-_--_-_--_-_-_--_-_--_-_--_-_-_--___--_-_--_-_--_-_--_-_--_-_--_-_--_-_--_-_--_-_---_-------------_-_--_--7S 23.. ECXHPEEMRIICMAELNSTAANLDCSOUMPPPOLIUENSD---S--------------------------------------i--------------------------------------------------------------------------------------8-9. 333.331... 33A3 .. EOSSPtRaqnaEuuaPntfmAlidCwRaoAAernnTIdIstSOs__aN-__-n_-O_-d__-F_-_S-S__-u_T-_-_Ap_-_p-N_-l__-D_i-_eA-__-s_R-_--_D-_--__--S_--_O-_-_--_L--_-_-U_--_--_T-_-_-I-_-O-_-_-_N--_--_S-_-_---_---_-_--_---_---_-_---_--_----__--_---_-_----_-_---_-_----_-_---_-_----_-_---_-_----_-_---_-_----_---_-_---_---_--_-_---_------_---_--------------------------------------------.-----------1199900 3333.31.. FSotortcikfiRcaottiaotnioSno--l-u--t-i-o--n-s----TM---.:--------.-----------------------------------:------I--------_--------------------------------------------I1I1 3 3 3 . Calibration Staadanis-...---- ------------------------ 11 - 4. 4M.1E.THFlOoDw_D__ia__g_r_a:-m------------------------------!---------------------------------------------------------------- 1122 444.334... 44SBSa4a4am.mt31cfp..lhlbbLSSPEeeivrxtraeotumrcrpEae_axsc_ntsr_tdia_snUc_cot_rni-oi_n--n_--e----_--E_----_x--_--t--r_--a_--c--_--t_--i--o_--n_-----._--.-._---.--_---.-._---.--.-_--.-.-_--.--.-_--.-.-_.-.---_-----_---_-----_----_-----_----_-----_----_-----_---_------_---_------_---_------_---_------_--_------_--_-_---__--_-_---__--_-_---__--_-_---_-1111132333 4 3 . 44Q34u.3a1..niLSiiFCaBIiMioCSno/lM-u-m-S--n-S-C-y-so-t-ne--md-i-t-ai-an--ad-i-nO--g-p_-e-_-r_-a-_t-i_-n-_-g-_-C_--_o--_n-d-_-i-t-i_-o-_-n_-s-_--_----_-_-_-_-__-_-_-__-_-_-__-_-_-__-_-_-_--_-_--__--_-_--_--111444 4 3 3 . Calibralioo Carre Procdtes-----------.----------------------------------------IS 444.337... TPa4Bec3kc3feRcp.SetQmaSCnaanUcnnecOEitCeaDCrAFirtOnietRarelAiyraNsi-iaA-s-_.L-.-_.Y-._.-S.-._T-.._-S.-._-.-.-_-.--._-.-.-_-.--._-.-.-_-.--._-.-.-_-.--._-.-.-_-.--._-.-.-_-.--._---_---_---_---_---_---_---_---_---_---_----_--_----_--_----_--_----_--_----_--_----_--_---_-_--_-_--_-.-11118677 56.. SCAAFLECTUYL_A__T_I_O_N__S________________________________________________________________________________________________________ 1198 ExTsmRcwanti Exygen Research P*jt3of43 Page 142 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExygesMetbodNo:BzM-023-071Reviiios 1 LISTOFTABLES * . Table I. Recovery Summary of PFHS in Rat Liver and Serum.-- ................. ....20 Table 2. Recovery Summary ofPFOS in R it Liver, Scrum and Urine -- -------------- 21 Table 3. Recovery Summary of FFOAm Rat liver, Scctnn and Urine------- -- ----------22 BxygeaReKnch Exygen Research P*jc4of<3 Page 143 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyfanMethodNo:EsM-Q234771tarital LISTOFFIGURES ' Figure 1. CaXbntian Carve fo rP M s................................. .............................................. 23 Hgure2. CeHhmxsithnve for PFOS..........-- ....................... ........... ..........24 Kguie3. Calibration Carve far PFOA------------------------------------------------------------25 Hgare4. Representative Chromatogram of*0.1 ngfaiL Standard CnaainhigrFH.s....... -- -------- ---- ---- .......................r..........._26 Figures. RepreaenmivcChrontrogramofa0.1ng/mLStandiidCoenammgPFOS...26 Fignre 6. Representative CtaeBMPOgnm of a 0.1 agfaL Standard Contiming FPOA..27 Hgare7. Representative Chromatogram of a 0,5 ngfaL Standard CcnttimogPFH5...27 Figure 8. Regararnnitive Chmmatogram of a (X5 ng/mL Standard Containing PFOS...28 Hgure9. Representative Chromatogram ofa(XSngfmLStandardConttmingFFOA.28 Figure 10. Representative Chram atognm ofiS.QngtaL Standard Contiming FFB 5-29 F.gnre 11. Repreaenlative Chromatogram of a 5.0 ngfaiL Standard Containing PFOS--29 Rgnrel2- Representative Chromatogram of a 5-0 ngfal. Standard ComainmgITOA -30 Figure 13. RFFepHreSK__c_a_t_iv__e_O__g_o_m__a_a_g_r_a_m__o_f___R_e_a_g_e_n_t_B__l_a_n_k_S_a_m__p_l_e_A__n_a_ly__z_e_d_f_o_r___30 Figure 14. RPeiOpreSs_e_n_tat;_iv_e__C_h_r_o_m_a_t_o_g_r_a_m__o_f_a_R__e_a_g_e_n_t_B__la_n_k__S_a_m__p_le__A_n_a_l_y_z_e_d__fo_r_ _ 3 1 H gurelS. RPFeOprAes_e_n_t_a_ti_v_e__C_h_z_o_m__a_to_g_t_a_m_o1fTMa R-e--a-g--e--n-t--B--l-a-n--k--S--a--m--p--l-e--A--n--a-l-y--z-e--d-^fo_r_ ,,31 Fignre 16. RepresentativeChromatogramof a Control Liver Sample Analyzed fbriE H S______________________________^-------------------------------------- 32 Figure 17. RfoerpPreFsOenSt_a_ti_v_e_C__h_r_o_m_a_t_o_gram of a Control liv e r Sample Analyzed 32 Fignre It. Repraeaative Otrotmlognm of a Control Liver Sample Analyzed far PFOA---------------------------------------------------------------- Figure 19. RfoerpPrFesHenSt.a..t.i.v..e...C...h..r.o..m....a..t.o..g..r..a..m....o..f..a...C...o_n.t.r..o..l..S..e..r..a..m.....S..a..m...p...le...A...n..a..l.y..z..e.d Figure 2D. Representative Chmmerngrara of a Control Semto Sample Analyzed forPFOS..___________________________ 33 34 34 Fignre 21. fRoerpPrFesOenAta--t-iv--e--C--h--r-o--m.-a-t-o-g--r-a--m---o-f--a--C--o--n--t-r-o-l--S--e-r-a--m---S--a--m--p--l-e--A--n--a-l-y--z-e--d----------34 Fignre 22.. RfoerpPrFeaOeSnl_i_ti_v_e_C__h_ro__m_a_t_o_g_r_am___o_f_a_C__o_n_tr_o_l_U__r_in_e__S_a_m__p_le__A_n_a_l_y_z_e_d_______ 35 Figure 23. RfuerpPrFesOenAta-t-i-v--e--C--h--r-o-m---a-t-o--g-r-a--m---o--f-a--C--a--m---r-o--l-U---n-n--e---S--t-a--p--l-e--A--n--a-l-y--z-e--d----------- 35 Fignre 24. Rate1p0reasegnfgtawtiivthe CPFhrHoSmat_o_g_ra_m_ --of.a...C...o...n-t.r.o..l..l..iv...e..r...S..a..m....p--le Forti.f.i.e..d.................36' Hay* Beaesdi FlJD5of43 Exygen Research : 661 : Page 144 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjmMated No: EiM-02M7I Keviiios1 LISTOFFIGURES (continued) Figure 25. Rato1p0icnags/ngtiwtiivtehCFhFrOomS.a..t.o..g..r.a..m...~o.f..a...C...o..n..t.r.o...l..l.i.v...e..r...S..a..m....p..le__F_o_r_ti_f_ie_d_ __ ____36 figure 26. Ratfc1p0mngm/gtiwlivi*c QPFcaOBAM--flD--g--m--n--o--f-i- --Con..t.r..o..l..L...i.v..e..f..S...a..m...p..l..e..R...a...t.i.f..i.e..d...... ...........37 figure 27. Representative Chromatogram of a Control Liver Sample Fortified at 50 ng/g with PFHS-----........................-- ......................... ' V Figure 28. Representative Chromatogram of a Control Liver Sample Fortified at 50 ng/g with FFOS.........................-------------- ------------- -- -- ___.38 figure 29. Representative Chromatogram of a Control Liver Sample Forfifiod at 50 ng/g with PFOA------- ---------- .------------------- -------- --------- -----------38 figure 30. Rate1p0rensgen/mtaLtivweiCthhJrPoFmHaSt-o-g--r-a--m---o-f--a--C--o--n-t.r-o--l-S--e--r-u-m- --Sample Fortified ............39 figure 31. Rate1p0rensgen/mtaLtivweiCthhProFmOaSt-o-g--r-a-m-----o--f-a--C--o-n--tr--ol S..e..r.u..m....S...i.n...g..l..e...F..o..r..t.i.f..i.e..d............_39 figure 32. Rate1p0rensgen/mtaLtivweitChhProFmOaAto_g_r_a_m__o_f_a_C__o_n_t_ro_l_S__e_ru_m__S__a_m_p_l_e_F_o_r_ti_f_ie_d_______ 40 _ figure 33. Rate5p0rensgen/mtaLtivweitChhProFmHaSt--ogr-a-m---o--f.--a Co..n...t.r.o...l..S..e..r.u...m....S..a..m...p...l.e..F...o.r..t.i.f..i.e..d...._ .......40 figure 34. Representative Chromatogram of a Cocttol Serum Sample Fortified at 50 ng/mL with PFOS------------------------------------------------------ .----------- 41 figure 35. Rate5p0rensgen/mtaLtivweiCthhProFmOaAto..g..r..a..m....o..f..a...C...o..n...tr..o..l..S..e..r..u..m....S...a..m-ple .F..o..r..t.i.f.i..e..d...............41 figure 36. Rate1p0rensgen/mtaLtivweitChhProFmOaSt_o_g_ra_m__o__f_a_C_o!-n--t-r-o--l-U---r-i-n-e--S--a--m--p--l-e--F--o-r-t-i-f-i-e--d------------42 figure 37. Rare1p0rensegn/mtaLtivweitChhwroamnAatogram of a Control U_r_i_n_e_S_a_m__p_le__F_o_r_ti_f_ie_d________ 42 figure 38. Rate5p0rensgen/mtaLtivweitChhFroFmQaSt~o~g-r-a-m---o--f--a--C--o-n--t-r-o--l-U---r-i-n-e--S--a-m.,-p--l-e--F--o-r--ti-f--ied............... 43 figure 39. aRte5p0rensgen/mtaLtivweitChhProFmOaAto--g-r-a--m---o--f-a--C---o-n--tro..l.U....r.i.n--e S--a-m---p-l-e--F--oi--rti--fied _____43 ExygenResearch Exygen Research : 662 : P ip 6 of43 . Page 145 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjcaMethodNo:ExM-Q23-G71Rcruxxi 1 r \ L SUMMARY T(PhFisHrSeXporFtedzeSooiolscosomcaestehsoadlfooafsnlEuiy(PiiFsOfSor) maniddigPserorftaPdnefctaafclosoorhoeoxcitnmeottki:lfaAnditde (PFOA) in Rat Liver. Serom and Orine. Reajdnea of FFHS, PFOS and PFOA are extracted from cacti m alrix wife ctalaaicon-njonaatolednymdTistiiipiowsflnriuiiaeVtshhdiend2Cg b1mTsy8ohuLellsitqoioapulfcilideedmtrpoceehhanthraciotastdimenoloenacLtxmoetgxrorQatnarcuapittacihootnyrcti/iintfs(agiScn(aaFdMdtEiadoR)nenMdcmao)ar.tftosrsFidFwsgpHeae.tScet,rrAoPamFnnaOedltySrytleoa(anLrdedCesdi/PdMFnoSOena/AMtoaStiesa) Tmheethpordopwosheidchlimgiviteosfaqdueaqnutiattaetiorenc(oLvOerQy; athcceolrodwinegsttfoecEtiPfiAtatigounideplienreifsi)edfobrytthhies method is 10 ng/g (parisqw-bUIian) each for PFHS, PFOS and PFOA Tanhde twbeiollsebtiecaaltlimleaitstofgdreetaescetriotnha(nLQ3D)tiwmielsl btehebalseevdeol notfhenosiigsen,albtaoanedd aoenratthioe icnofrirnemspeonntdastitooc0.s1ynstge/mmLu.sed. For all analytes, the lowest analytical standard This method was developed using tat liver, serom and urine. Typical percent ' ' rccoYcrins standinl deviations (at 10 and SOnglg) ate shown below; PLoevrtei3Sl10(c0aatriot)o PF19H18S3%*ReL3c9io3.v9v*e%erCry(ii*n*33R))at LFecvretlif3(1in0c0gii/imonL) PE:lolHlw*SR&9Se.ec6rooa%vr*ce.r(fyci*i*n33R>)at Level PFmORSUReLciovveerry FarLtifclcradtioc PFORSaRtSeccoavmeryin PFORSaRteUcroinveeryia 3100 9868**81..55%%((ti**33)) (at5f1m00 L) 18280*9Z.81%*((ii**33)) 9739** 4L.27%* ((t*-33)) (Lee51^v00Eel) PFiOnARaRtelcivoeverry ForLtiefvtoetlion PFORAaRtSeccroavmeryin PFORAaRxeUcroinveeryin 9984%%32.1i*%(r(*t*33>) 5100 111117**A1.O5** (0i**33)) 8897%*22^J%*(<rt**33)) Rcherpormesaetnotgartaivmes csiaclibshraotwionnincuFrivgeusreasre4 tsoh3o9w.n hr Figures 1-3.' Representative ExystaRsansirb Psge7of<3 Exygen Research : 663 : Page 146 of217 MIN 313/023622 Exygen Study No.: 023-076 BzTgcnMethodNo:EzM-02S-071Rsvisioc 1 2. EXPERIMENTAL COMPOUNDS H e toctora for FFHS, PPOS n d FK5A n e given below. FFHS Moiecoler weight 399, u hows FFHS u rap p fo lt the ponmiam aJt(CjF|3SQ)'K*).mr)lcca!r weight <3S paMgroomkscneldarHwaemigeht 4R9a9f,haxsxsohoowcms iaulfccite PFOS U uppHedm tfae poassum n it (CtFoSOaK*) molecoier weight = 538 pMChroonlcnaei.ncililrNweaigtht = 4P1ai3a,risnschifolwoosroorminic Acid ExyicaRocerch Exygen Research : 664 : PifolofC Page 147 of 217 MIN 313/023622 Exygen Study No.: 023-076 S x n a K ited Na EaM<B3-<!71Renata 1 /--> 3. CHEMICALS ANDSUPPLIES . . . . . . 3 X Cazm cog__________________ ____________________________________ Chfmtrai_______ Grade_______ Sparer______ Catalog No. MAmetmhaonnoilu(mMAoOrwHm) e , RHePaLgeCnt EMJTSBciaeknecre MX0509467-S0-11 WAcaetteornitrile Type I u vLC EMBaSycgieenoce AX0N14AS-1 Type I water electrical resistivity, mronnum of 16.67 MQ/cm at 25 C, from a lafcccneo WwaproTM workstation. 32 . Standards SWtnafnlWdatnrSdhtfcjrwrwwiTfnrurtf. P(PaFfhHwSr)ooctanegulfaaaie P(Pmf]Of<SW) if^v>pyy^nnir. AddffFOA) TCH Number S&036 SD-018 OLSo3t1N6Do:O Purity (%) 89493.969sat8rla6li.ig9shotmcehrasi,n 96 Source 3M 3M ACldbreimch 33. EQdFMZNTANDSOFKJES Equipment B12al5a-nmceL;LaDnaPlyEtincaarlro(dwismploauytahtbleoattslet0s.0001 g) TDiissspuomsxazbelexglass micropipets (50-100 & 100-200 pL) WScrriysatDacRtiCon5sChapkluers Cenaifnge 5105 mmLL ddiissppoossaabbllee ppoollyypprrooppyylieenoee cceennttrriiffuuggee ttuubbeess Visiprep vanmm mardfold S02PS3r-fesma6pnf7rLPi9na5dArkreipVai|prs*hHc*e6P*qLwuCip(vmlvginae)llnrtKtmCi1^t8f(rcfcacatAm#mi3d1Wg8cey1sl-i(3np4dm0e0rs)#. WAT LHHcS8B4CtPya84s/Lpn4pM0d4oeC10-srS7aa1oP/-lb7M4rool-o0enm4S0etd0pi)0rradbo)(nrUfpodr-ptMHie-nitPQncgL.uAg)CanDradar)ydcsoctelaumrmtarindg(4e holder ( p m mm) (part# Supplier NMaelgtteleore DBruumrreTmlelokSnmcdiae(VrntWifiRc ) SDWVVuopWWapemelRRcsno!. vHaerwioluestt-sPuapcpkliaerrds various sippliera Keystone Socadfic Keystone Scientific As described in section 43. ExygeaRejmch Pap? of43 Exygen Research : 665 : Page 148 of 217 MIN 313/023622 Exygen Study No.: 023-076 EzyjcnMethodNccBiM-023-071 RevhioQ1 1. 2. tIunboersd,eporiptroTetaetevfsloo.inedt-cfcl.oy)n*its*ahmiWginhatlrtyiiforlWncc,*ofenoemriufrspaeirpoirfrriidw.irsiptonshafbtiilledlfaKb*wHsrrc (ccntamm, 3. IpCtreemrsteaaniyncbeloeotnfTmeccmoensbstamamrtyihntaoavncethsbee(deecsnp-ffeeoceuianslodlyltvoFebnFetOt uA(na)csuebtioytanbLiltCeril/feMo,rSmu/sMeet.hSanboelf)ofreartofeee. 4/ S. UwEqsheueinvdpaislreepnpotasmraibnalgteesrmitaalinscdrmaorpadtyspbeatnetdessusbaomsrtiptpuleitpseedftofteorser xtthtoroascaetliisoqpnue.octi.fsimednidnatxhdissmoleutthioonds.- 3A* Soum oets (1) 20.15z4oMg ofmsammnauriimumu aacceetatateteandindiswsoaltveirngiins I Lpreopfawreadter.by weighing (2) Hypercart? filtered type I water is prepared by filtering type I water tBafhicleretofetoourrengedihtrtuyailspeHee,,yItpwhweearnaCsthae-rrdfce2ghe5ougtomeudLafocrdoeoluufcsmaetryntiprniuedfsgeIineegwewxaattirHefaercP,tL-ifo2eCn5e.spumRbmeLeppgeioanatft c-fH2eoe-lPl3weLczCatnsiLnhgg/narxafftmdeeee.r filtering -2L of water. Nmoa tye :b eT hpereapfaorreedmaesntkiomngedaesxafemepaleppi sropprorivaitdeerdaftioorsgouifdfaeneces,oallvteenrnt attoivesovlouJtaemacres 3 3 . Pszpaxatn o r Standard Solutions Anal1y.ticCs!shstbarnsdtiaorodsSatraenudsaerddfso-Tr thheresee sptuarnpdoasredss; arc prepared in methanol sudare ' 2. LcduoesantebedcoremrtnaoittncroaeartyhiaobnntCasslfoyecntofitcerrroaeellrsrepeSsocppnooldvincieensrsegy.ot-foLfeatThebehodereLsateOteocQrtfyooracrutnoisdnfietcrd5aoxitlinoLstnpOhsueQraasnraaeanlrdyepsapirserr.eeppauarsereeddd tiaont 3. Mecscafolifmaneettncrpritotlx'elossSmfreapfaqteitkuaerieexksssa.nt-m.oTwphlneecsmoeanftcorerixtnitforiacntaiatoinnoan.lsyMtaircaeatrlpirxreecsppoavirkeeedrsybaayrneadupaserdednpgtorieneptvaoarelfudeaeateftifefeelede Tprhoepaonrtailoynsst omfasyolvuatreytofeseoalvbesnotluatrec vmoaliunmtaeinseodf.fee standards as long as fee correct Exys*nRoecda ** o ore Exygen Research : 666 : Page 149 of217 MIN 313/023622 Exygen Study No.: 023-076 ExygenMethodNo:ExM-023-071Rerixkm 1 " 3S1. Stocksedation . # PPpiLsntoPrDaezObspPieltAaEeyprafesborboayoirdnttaetdwilmifer1vsina0ig)dex0iucari-miearmneslugsLamstootrvoyuopcb,telkeusr1ami0osslottedomltcuorroeigtfcnidoootfneniflnssaeets)ysakatceasrha.ne1rdf0afTrn0rdiohgaQmpleeuyrgttasdfeittrcoontaorcLldkas1aftt0tsoaeo20nrloduxPCfatsFiprLodtrBnoewS(spc6i,ato(*rhiPraCnrtPmeioOc1actn2tnSeh.5ddt-oamaf&norLdeIr 3 J JZ . F o rtific a tio n S o lu tio n s a. Prepare a mixed fortification standard at 1.0 pg/mL (1000 ngiftnL) of PftFrvHySf,foP1FutOioST>|ainindtoPFaO1A00-bmyjLadvdoilnugm1e.t0rizcaL of eachborifntghuep1t0o0vpoglufmaLe b. wPriethpamreethaamnoixl.ed fortification standard at 0.1 jig/mL (100 ngfrnL) of ExamPfoFprltSeifS:ic,oaPntFeioOhnuSsnoadlnuredtdioPmnPOtkorAo1f0bit0yemrdsiLlouwftitinhtghe'm100.e1.t0hjimagn/LomloLifnstoahlevuot1il.ou0nmnse#ptrimkiceLdf,lmainsiktxo.ed1 g of liver or 1 mL of serum/urine is equivalent to a 10 ppb (10 ngfrnL or og/g) fortification. ^ bSottotlreesa)Qatfb2rtCifitcoat6icaCi sftoarnadamrdaxsitmatuimonpseirnioadreoffroigneerayteoarrOfnro1m25t-hme LdaLtDe PofE pnerecpeassraatriyo.n. Note also dial additional concentrations may be prepared if 3J3. CalibrationStandards LpthrCee/p0Ma.r1Sedj/iMgafStc0acL.a1lm,i0bi.rx2ae,tdi&ofn5o,rstLtiaf0inc,da2atJri}odnsanscoodln5ut.ta0iionnnign(/gsmePcLtFiioBnnSm3,.eP5th.F2aO.nbSo).lavnida dPiFlaOtiAonaraef Tprheepafroeldloawsinnegediesd.a typical example; additional conceotratioas may be Initial Cone. (n g1f0m0 L) 100 1520..000 L0 V(om5ta.L0m) e 1112L000.O0...000 Di(lum1t0eL0d) to 111100000000 100 Fi(nnag52<00lI/D..DmCu205aLn)c. Tbohtetlessta)nwdhaerdnssmtoareydbreefruigseedrafteodr (aatp2erCiodtoo6fCon).e year (in 125-mL LDPE ExygiaSfiseaich Page 21of43 Exygen Research : 667 : Page 150 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjenMethodNoeBxM-CE3-C7lReviwo 1 4. METHOD 4 J . F lo w Dug kam The Sow diagnm of tbe method is gives below, followed by a detailed deunption of each step. Method How Diagram Weig(hfoarptipfryosparmiapteleasndyexsrritg!noaftelidvaesr omramtreixassupreieaspparnodplraiabtoeravtoolruymceonotfroselrupmikoesr)urine Add watef to Ever for a finalvvoolulummeeooff2100rp4im?L-L,haodmdowgeanteizreto scrum and urine for a final Remove 1mL and adld 5 niL of ACN, Sluice 4O n trifn g r. D ean: supernatant into 35 mL of water Load onto co4nditioned SPE Elate with 24m'L methanol LC/MS/MS Analysis 4 2 . Sampl* Processing Fdroyr .Eicvee.r saTmhepnlesp,lapclaecesafmropzleens sianmcpolenstaiinnearsfoaonddprloecaevsesoorpaenndihnofmroozgeenniszteorwaigthe ostvoerrangieghbtetloowallo-1w0foCr CunOt:ilsnrihmHemaorflone.xtrSaectailona.nd pAlaltceerntahteelysa, mifpledsieirne frios zetnn iasnensrduufmtfhiceaiensndatmuaprmilneoeucnsaantmobfpelseausms.epdHleaoswseulvepesprs,litfehrdao.nzeSNn gos)e,sratuhmmepnlaennodprpuorrcoiencseessissnaignmgipsilsenseneemdceuesdsstafbroyer allowed to completely thaw to rocn tCfflpcateto before 01c. r' Exyjca Research Page 12of43 Exygen Research : 668 : Page 151 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExygcaMethodNcsExM-C23-071 Revision 1 r s 43. Bitch S rr up . #. a. A btxch of sample* should not cootam more than 20 field sample*. b. Ecbolaancnchkenbutarstacitnhiognoscfos(ntaytmrpopilclaemlslyaatnr1ia0xly)aznaedndd5m0towsnotg/ignmcafluotrdriexlivacteornletoarrostlnsgo/nfmoerLtcioffinoetdrrosaletr(umkmnetohawondnd urine) to verify procedurallewYcry for the batch. < c. kAAndtodlweitanisotncoaolnnsecaefmnietprlaldetissoainnmtaphnleedhincaatecrrahicemhdabtyhafrtcolxhungmhhuetsfhtoeartlpisfroioecbdee*dfsuerpeatroatevleyriffoyrtrieficeodveartya. d All samples require daplicaie injections. 4.4. Sample Extraction 4 .4 .1 . U v e r E x tra c tio n a. Wforetiigfyh, 1ifg aopfplriovperriastaem. plNeointetothaa5t 0azltreiLrndaitespowseaibglhetscenotfrifliuvgeer tusbaey anbdo bcd... e. HmTCmAdireoedisannmapdnutsorswoutsifegfraeasueebtrdgneolerei1ndzteeaoctmpeuswtenbahLnrmteeridssiospiftnauflasemggtctueohtpis-neloi3tennut0shfbgaso0eehmar0ssa.ahpakmmlrfeeiprnAp.imaeludelisidvisufnioozzSglrceutmam-afvoeLdar5oii--lsofapm1bf1ol0uiseAnaumfnbCaotlnLNeertseup.sCaeinpa.dCeptattsierghehafinutkllteyloy. fo1dr5ec-me2nL0t * f. Lswstrueoaieppateepdsrreen.tdchatteoiaonsnnattmhi4nep.t4Sloe3Poa.E)n.5tco0oDlaumicmsLocnnadrdadiitsttpithohoinseseapdbcollSueiPnacttEee.nctoriAlfuunmgyne tube and add 35 a(nfoarlyctoenrdeistiiodnuiensg mL of wdeitlal ilbse, g. fa. ErAalTnua.tcelyewznetintshfaum2gpemlmeLsbuoesfimngetehleacntorlo.spCraoylleLcCt /2MmS/LMoSf. elute into a graduated 15 L Seram4 .4 2 a n d U rin s E x tra c tio n a_ Measure 1. mT. of serum or nww mpT* into a 50 mT. disposable cutfespne.tiyrwifutgieHtiuirbiney.amndyfobretifmy,eiafsuaprepdrodperpiaetned. inNgootec that alter7n/a*t.eavvsoQluambleosfoorf Exyjeu Research Page 13of49 Exygen Research : 669 : Page 152 of217 MIN 313/023622 Exygen Study No.: 023-076 SxygenMrtfcodNo:2x2i-023-071 Rrmrioo1 r * b. Afbdr-dImwmu enrstco. nTmiepnlceo&ntiianufieiwa litvii3ttttmpseddi2i0oaleLctiConp4.d4j.1ir.tly nd vortex 4 .4 3 . S P E C o k a rx . C o n d itio n in g Place the SPE columns on the vacuum " nffold, Condition the tSbuhsyPeiEn~cgoc5vloualmmucmunLunumosnfabaiwytdaeapdtftes.lroi.DfwfiiTgsrach-taeerd1ow0afal-msl1hwLedasosrohmfpem/sys.eetDcbheaoonrpnomuolltatlheaydlrblooeutwhgarhltolhuotehgwceheocdtlohuthemormnSpePntotEsfdotcrhloylrol.ouwumgehdn 4 5 . Q d x ttta t o n 4 3 .1 . LC /M S/M S SysU m a n d O p era tin g C o n d itio n s IMntaesrsfaScpee:c: CSoofmtwpaurtee:n Mcromats Quattro i ntima (McroDMBa) HEkacrvtraorsdpicnafyla(tihofnicpToommpas(sH) arvard Insmuncate), for tuning COMPAQ Professional Workstation AP200 Window* NT, M mlynx 33 HPLC: HewlettHPPacQkauradt P(HumP)pSeries 1100 HP Vacuum Degasser HHPP AConhtoirsnanmOplveern Note: Atchoe4nptxaumrlgOinemavnmatlsvbteyhpametrmdcbaazyebfboderreiontphtehinesagmmuoapbrldeilecinapjrehtrcaitsdoegraepniesdn/taotttroaHctrhPaeLpdCaonsnyy-lsritenesemida.ufteer HPLC Column: Genesis C(Jones Chromatography), 2.1 mm x SOmm.4^ Cohirnn Temperature:35 C MbjeocbtiilceoPhVaosleu(mAe):: 21m5 pMLAmmonium Acetate in Type I water Mobile Phase (B): Methanol TItty. 0z5..o00 a991a000 9.0 119445.50 910000 20.0 90 31199111L100000000B00 Flow R a0000000tt5555555ftnL 'nAO 05 ExygeoRateareh Pag# 14of<3 Exygen Research : 670 : Page 153 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exyjcs MsthodNo: ExM-G23-07I Rcvsoql IfptreormmfoarydmibfafenercneeencteCmssoarrtmyofntfolaerawndnijBtuhrs*dt(itKfhfeeeyHrsetPdoLndCeimBgeerntaasdaiioQenn*Ct (iuen.egotTrcd2.).e1rcoxtou3lod0p)btaiemndtiozaeeldsio,nspctroroulvumimdeenndst equivalent chromatography is obtained. iHULfflotieaa* Azute PFHS PPOS PPOA Mods TftaraypMOTiwrai Negative 399 -* 80 Negative 499 -99 Negative 413 -369 Approximate --8.2 min. --88..86 mnainn Tmastnhaoanebldyirlateeirtcdepanshltuairosuseneni,etnitacmcsl.ueldsoDenmdrgiaafyattsidvnitaehrreebyte,edgnoritinnifotnancindotgianmyatinentdsooe(dunapdtyhtorobofautshgies4h,at%dnheae)plyieestnnicadtiacirnlcegeruaponntan.abltylheseiwsbiaathntcidnhtahonef Neqoutiev:alAennt.alternative LC/MS/MS system may be used once demonstrated to be The mass spectrometer is toned for each analyte by infusing a - 1.0 pgfaL sotfanmdoabrdilesoplhuatisoenc(oantt1a0inpinLg/m50in%, umsientghaannoilnafnudsio5n0%pu2mmpM) viaamam*oTn*iuimntoacaestatrteeaimn apwnaardtaemtrheiettne0rtsu2naemrdeLfs/oamvrietnhdeflaopswroadmtuuecnt.eioEfnial.ec'.hOaInbncaieslythtteueniisensifntirliuetimsiEseynthttoeisnnetuudnsfeeoddr,dtthuhereinpogaprteriomnutitziionende analysis. 4_5L2. C a lib ra tio n C u rve P ro ced u res a. IL(nrCajen/McgtiSnthg/MefrSso.ammteheallioqwuoetst(lbeevtewlesetnan1d0artdotoSOthpeLhi)gohfesetalcehveclaplribepraatrieodn),sitnantodathrde b. Use weighted linear standard curves for quantitation. Linear standard curves pecaoqareueluatclikiugvelearaanlteriebenoryatna)teousvdofsefirffntsewoguersacraeaecaalasicbylhairsbpatareptanimrotaoinl.opynrcAtieuanrstbvetyaeycn.oadluHiHantrlboeidrewaarrtecirvotoeeneagsrcstr,e,tetanshnmtserdiaoaattynoirodtanubflseoniuunusengmixdnc1bgtl/oeuxrdMboewedfaeascisagfsllrthyiaobtntmiirnxsatgtiitc(ohooanefrl ssttaannddaarrddss itnhjaetctmeda.y be excluded must not exceed 20% of the total number of c. The correlation coefficient (R) for calibration curves generated must be 2astp&apn9rd9oa2prSrdiast(oeRr2tsh2tee0p.rs9e8lem5v)ua.snttIsbfeetcoatlfaiksbaeramntipoto!n arsedhsjouuulstltsd ibfnaesltlrreuoamnuateslnyidtzeeodpt.heerasetiolnim, itsa,dthtehne Exygm Research Pago15of43 Exygen Research : 671 : Page 154 of 217 MIN 313/023622 Exygen Study No.: 023-076 SsygcaMethodNo:EiM-023-071 KfiYincnI Typical calibration carve foi FFHS, PFOS and PPOA'csn be found in Figures 13. 4 S J Sam ple A n a ly sis a. rIencjeocvtertyh,econmtreol,aelitqc,uiontto(bthetewLeCen/M1S0/MtoS 5sy0stjeimL). of each standard, temple, b. Standards corresponding to at least five or more concentration levels (starting with the LOQ levd or below) most be included m an analytical set c Asent feonltliorewseedt boyf ccaalliibbrraattiioonnssttaannddaarrddsssihnoteurlsdpeberseindjeacptepdroaxtimthaetebleygeinvneirnyg5o-f10a ' sabmaletumgesripntnlnbaeitesnivtgie(rt,eaonfaidnrasactectanotnhtuidenrelteanssfdteoitronjfoeaafctscsiaoeamncliobpinnrladeatsisoseanetmt spItonlaefnesdieetaxhtrtedrrascctmaesdaey, cstabalenibdiranarctdihosx)nf.esdtanAadtsartahdnes d. The concentration of each sample/fbrtificaticn/ccntrol is rictenni neti from the r \ rRrsnaeteeanscnspgeudoeslan.strsasdermysc,asuydhriovbluueetl,edqbubtaharsneaetcdisktaaemmtterpidelesuthpspeottnoopseeg1sai0vko%efaoatrheueratesrsiodepsfeoindtehsuaeeechcwfuoiarutvnhneaidnlbyitytnhe.eerxstrrtThaabnpseodalamasrttdpiaolncendu.sarevrItdef. e. Fortification recoveries falling within 70 to 130% are considered acceptable 1 Samples must be stored refrigerated between 2C to 6C until analysis. g. cSaonanmaclpyeltneetrraeitntieonwntihooincfhtthiemeitehcaewlribnilrolatbpioeenarkessptaoanrrdeteaddredatsescatrNeedDdoe(rtnepocettaedkde-taeltecsttbesdet)h.caonrSr.teahsmeppololnewdsienisngt lwceasilnsibdtrh)aaptnieoatnbksestaLinrOedQadrdeatsnewdctigeldlrebaaettertrehptehocaronterodrerasespqNounQadli(ntongotatbnqeaullaoynwtteiefrsieattbcelonen)ti.coennttrimatieonthoaft tahree T5i1n00hsenaengandn/ngan5loy0afsnnPidsgF/puOgeriSronffoear.nFmdFTeHPydpFSidOciunaArlirncianhgtrmftoihvmeefarmi,tvoe1egtr0hraaoanmndddsd1c5e0av0neanlnbogdep/m5mfo0Leunnnotgdfi/nimFacFLlFuHiodgSfeudPirneFf*oOs4reStr-iu3faim9cn.ad,t1iFo0FnOasnAadt ExygenRa cgcfa Figs 16of43 Exygen Research : 672 : Page 155 of217 MIN 313/023622 Exygen Study No.: 023-076 ExypnH a tat No:BIM-02W71 Itoruxx 1 A C dPlxyc* Cbttzria The following aiBd> most be met to ensue the presence of FFHS, FFOS and PPOA: 1. Tp4a9hr9eendatsnmoofaf3mo9ro9FgaFrmaOmoS,mfoaornadPtFasHhdoSaw,ugaahdpteaernagikohontrfaratic3(ie6a9nEgah9stm9rraiforrmonmaftraeSeOpr,aaranemnptarfoerfon4mt1o3af mmiferFiOA. 2. Method a blank N tnln rrtntf contains' the naontaClyotnetaailnLaenvaellystgeraetaLteervtehlsagnrr1jt0trnygthfaaLn tbo LOQ. If , tibca a new blank ample most be obtained and the entire set mint be re-extracted. 7Ramd0ceeac-tct1eeorrp3ivmxt0ea%irbnsilpeeesioiklffoiermfetho-iceteuosxit,nrsttrtirahdkoceenltoies7owpn0nitn*kiir1seev3sw0asal%aeunrterdaossn.mfhtoesaduaItrfm.lidxpalbsecpesoiknseehvtrsoaoluQluldafstpaebnidekye)rbemyf-aeuldxlssittembocauetnttsbdaiel.dytewsAtetnehtnoye ABHmoonawyostrecnTvaoleeitrbs,etr,xtahmcteieoaetnyodtsba2tale0nn%eduxamocrlfdbutefdhoreeudotnoffdtrcaoatloHmnbbutnehmtefabicaseantralocstitufsaltsnaitcdtaiaanolrdndoasuordttfEhstaehintrejbcecoycautlueilbsddir.nbatgeiotehnxec.clHuurudvgeede. The correlation coefficient (R) for calibration curves generated must be 2ast0paJpn9rdo9a2prSrdias(tcRer1stthe0ep.9rse8lm5e)vo.asntItbfseceatbobfcsaatmicpulersessuhlotsuilfndislbtlreuomrueteasnnidtaelHytpzhemeds.etfolTim*, iatsn,dthtehne Retention tunes between standards and sample? most act drift mote dun w4ith%in awnitahnianlyatnicaanl arulyntitchaelnruthne. seIft mreatesnt tbieonretalnmarl.ydzreidft. exrerds hi Emit 4.7. PlSJOSMANCS CKZIZSIA Tbheahfseonrfoeotldlboieewecnionumgsmetwdeofnoccrertmihteiesrnimtaoemfthuaosndta.lbyesipse, rwfohremneudsionngcaenaisnsatrsuymsteenmtatsiuointasbeitl-iutypttehsat,t RFiurnst Ca risttearniodnar:d solution on LC/MS/MS corresponding to the estimated LOQ /--n aa(tp1tnp0ldernocagphs/trami9na:Lgt1ee),).icinnosmmtrpuaasmriexednattnoodpaoerbreataatgiinnegnatpsbailrgaannmakle.tteoIrfnstoh(ioissrecirnraictteiroeriafosonerctthhaieemiaonncjaeblcyettiemonterttvz,otoslpuittmiiomenL,orieff HxyyaBcjccch Paps 17of43 Exygen Research 673 : Page 156 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjenMethodNa BxM-Q23-071Rcvinca I RLSGeOoecnnQoena,radsoteCeptraoittcfoeanslttoiahbnner:daathiroidgn*hcoeusftrfvicevoefnoocrerdnmiteroaartienoacnloynlteecveaenn*ltrd*atootibotbnae#ilneivnaecllsliu,ndefreaodrmreingartetohsrseiboeannlowawlyitsthhisea. ccroieteffriicoinenist mofetd, seatemrmpliensamtioany b(Re3a)naolfyzaetdlewaistth 0st5a8n5darfdosr itnhteersapnearlsyetde.. Once this 4 & Time Requeued for Analyse isOsntaanmanepdpppaleerrosrds,xoiinm1njecamcattenailotytrnai4xsk)ehwboaluiaslrnlesktt.a,okTf2eh2ae0lpaLspbaCromor/xaMpitmloSers/ayMttehcSlryooanu1ntg4raohhllyodssuiipesrsisok.afemsth,pel1esepmtrea(cptroainrxatatsiiopnniinkpegro2ac0neddfiue1rled2 5. CALCULATIONS a. UpeseakEqauraetaio)nu1sitnogcatlhceulastetatnhdeaardmocuunrtvoef a(n1a/xlytwe feoiguhntdedOnliangefamr Lr,ebgarseesdsioonn p a u n rten ) generated by.the Masslynx software program. Equation 1: Analyte found (ng/mL) (peak aredao-pientercept) x D F x aliquot factor DAlFiq=ufoatcftaocrtqbry=w1h0icfohrtlhiveefrin, 2al0vfoolruamereowmaasnddiluurtiende, if necessary. b. FEoqur astaimonpl2estofocartlicfuieladtewtihthe pkenrocwenntraemcoovuenrtys.of analyte prior to extractioc, use F q ra m w i 2 : . Recovery (%) *= [total analytefound (ng/mL) - analytefound in control (ng/mL)] nalyte added (ng/mL) X Note: Sifunbgtr/amcLt ainnaClyotnetrfooluinsdgirneactoenrttrhoaln(nLgO/mQL. ) from analyte found (ng/mL), ExygaaRMetrah Pap LSof43 ` Exygen Research : 674 : Page 157 of217 MIN 313/023622 Exygen Study No.: 023-076 ExyiecMo&od N a BaM-23-07lRcv$sk I c. `Uic Eqnjdoa 3 to calculate the amount of analyte found (in ppb) Pimarifm A u2yatalsl(l]rtorfljtaL)a trfipm*Marigfohtmd fCmCfTtnTLrrpIIyFvVniwfgnflr.1(asL) FV = volume , - ,* .Fleodsres srtpehpoaonnrdtthiinnegglopwaunrepasoltyscteeosn, rcseeatnemtnrpatiltoeiosnnitniomfwethhiewchcinaeliibbthereartrieonpnoosrptteaeadnkdsaarsadrsJe3aDdreet(ednceottetedcdteoedtrepacteteatdhk)es. tSthhaaemtcpaarlleeibslreianstsiowtnhhasinctahtnfpdaeeaLardkOssQwaraiennddbeegterreecpateotedrrteathdt atahnseoNcrQoerqr(uenasopltotqonudtahinnetgilfoiaawnbaellesy)t.tceornectcennttrioatniotiamoef 6. SAFETY Trhlaeahbanegoderlaanitnntosagrlyysbstceatofnoasdrhtaseo,rduspasldefarefnrtoyedargmdrleaiatnshgsgeeenst,mthsai,anstmdermegifaleooltrvhiseioansdfg.e.twyoUdrkastienagmsihneivefteusnmfaeol rhpoaroleldcsasutaatnnioddna*wrdeswarahinnedng ExygenBcaeard Exygen Research : 675 : Fife 19*43 Page 158 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxyjenMethodNc:ExM*QZ3-C71Revirioc L Table 1. Recovery Summary ofFFHS in Ratliver and Serran RecoverySummaryof PFHSInRai Liver 000222j0002111686S888444tSSSEptff*kcABC-- S 111000,lS S l-- Standard DAawvfaatniosrc JSSii1111S01218S6065 ECi 000222000S111a666m888444piSSSapelpDAkkEDF AratfSyttanAd55ad000rddadDA(envvjija/nstgi)on:; Paroant R1S69a0CS4o1ovary<%) ZJS Recovery Summary of PFHS In Rot Serum 000222000111666888222 SSSpppikkeBCA 000222000S111a866r88o822p2iSSSapeplDkkkDEF 111000 Standard DAavVtantiSoKn: AnahSrttaaAndd555ad000raddD(AnavgvtataarHaUgoan:: 111141001.72380 Prcaot Recovery(%) 1111M10021742 Ezygea&eanrdi Exygen Research : 676 : Pi*etf43 Page 159 of 217 MIN 313/023622 Exygen Study No.: 023-076 Emeu MethodNo:&M-0ZM71 gmHon l Table 2. RecoTtry Summary ofPFOS In Rat Liver, Serum and Urine ficovry Summiry of PFOS In Rja Liver 000222000111666T686444iSS8lpppIkkkDBCA 000222000s111r6668f88444e SS8ipDpofikctFE0 K iaSMtanUd111a00S0r'dMOAe'nvvsta-mgB'i:obrkc AnaStyttaenAd655ad000rddeDdAn(rvttgae/trftlao)gne:: PtrgtttR18t8690-85656t . (%' PuoMt R19888to0768JovMyfX) R*cowy 8wnmwyo( PFOS InRt Setun 0002220001116868882228SSpppkkkCAB 0Q02200s11166rtt88f222BeSSpBppkkkEDF 111000 Standard DAevviamtioonK: AnaStyttaenAd665ad000rddeDdA(envvgiea/rtmaiogLne):;. M68890689 Percent R11112e2122.c11802overy (%) RecoverySummaryof PFOS InRetUrtns 000222000111a666m888222pSS8tepppaiPktcACe AnaSlyttaenAd111ad000rdde-DdAe(PvfOiKatttaiQoLn#):! PercentRe49898c.16837o.veryf%) 0Q022200S911a166r8R6a22p2tSSSeppl(D*kkDEF AnaSiytatsnAd55ad000rddeDdA(envvgieafrttainogUne:; Percent R1777e8*80cJovery (%) SxygnaResearch Pig*21 of43 Exygen Research : 677 : Page 160 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjenMethodNexExM-023-071 RcrjxkmI Table 3. Recovery Summary ofPFOA in Rat liver, Sennn and Urine Recovery Summeryof PFOAin Ret Lhmr m plaP 000222000111666888444 S8SppdfkccBAC Analyte Addad (nota) 1100 10 Standard DAavntoniognc: Parcant It o w y fW ' 18061 w 88.11 tam piaP C00222300111486888444 SSSppf*kkEF0 6600 SO Awags: B ttndvd Devfetfcxe Paroant A aco w y <%) 888724 84 2i RecoverySummeryof PFOAin RatSerum 8 a a p ia P 000222000111666888222 8S8pppkkkABC Aaafyta Addad (ngrtnL) 111000 Standard DAevvteerdaogcac: Parcant Racovary {%) 111111867 . 1t-1E7 tam piaP 000222000111886888222 SSopkkEF * Anatyta Addad (ag/taL) 55SO00 Standard OAavvfcatriafegna:: Pareant Raoovary 111110725 111 ao RecoverySummeryof PFOAin Rat Urine a m p le ID 00022200011166088222 SS81pp*kkBCA 000222D00111666888222S8SpppIkkcFED Anatyta Addad (agSnU 111000. Standard DAavvfaeOraogoe: AntrtsAddadfogftaL) 60 5500 Standard DAavvtaartafagna:: Paroant ftaoovary <%1 S88168 ts 2 JS Paroant rtaoovary(%) ae 8886 2.71 ExyjenR*iearch Pagt22cf<3 Exygen Research : 678 : Page 161 of 217 MIN 313/023622 Exygen Study No.: 023-076 Eljfan MethodNo: BzM^23-071 Roviiioc 1 Exyjea Ketemcb Exygen Research : 679 Pjo 23of43 Page 162 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyxcnMe&odNo:E*M-Q23-07I Rernx 1 Figura 2. Calibration Curva for FFOS CCCCRuooauJrmepvbfofoprioocntyiuieopnnnedtt:oyc2UufoMrDcvnosiwBtm:e3c,:2vrOPi1nrFr1uatOgtJ]iObiSSorftEid:a0d.9u49d87o.0,9CW288o5ightbXF^ A>d*trans: Nono ExyjnoRJMBch Exygen Research : 680 : Page24 of43 Page 163 of 217 MIN 313/023622 Exygen Study No.: 023-076 Bujnn MhodNe BlM-OZW?! Rrrjcc 1 Figure 3. Calibration Curvefor PFOA ' ' CCCCRouceaerrsffvppefeofoicttnufiyesonpnendett:oyc1UfupnreDnv:aeeenaBt:or$ed,n1eOrPim2nrF7iaagO0fiiiA.nSo9:trd*Bc,xA0ci+.r8u39d*8Se79,54W.63e6ighting: 1/x,Aidetame Nane Exyjco Research Exygen Research : 681 : Page25 of<3 Page 164 of217 MIN 313/023622 Exygen Study No.: 023-076 BxyjccMethodNcr.ExM-023^J71Reviren 1 Figure 4. RCeopnrteasineinntgatPivFeHCShromatogram ofa 0.1 ng/mL Standard Figure 5. RCeopnrtaeisneinntgatPivFeOCShromatogram ofa 0.1 ng/mL Standard Bsyses Research Exygen Research : 682 : Pije 26 of43 Page 165 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxyjmMcdKdNo:EM^&071 Ktvioo 1 figure 6. RCeopnrteasineinntgatPivFeOCAhromatogram ofa 0.1 ng/mL Standard n u iw m u m .1min IO .--.-- v'-s Figure 7. RCeopnrteasineinntgatFivfettCohromatogram ofa 0.5 ng/mL Standard Exygen Research : 683 : Piie27oM3 Page 166 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyxroMldNo:Exi4-CZJ-C71 Roritwo l Figure 8. RCeopnrteasineinntgatPivFeOCShromatogram ofa 05 ng/mL Standard Figure 9. RCeopnrteasineinntgatPivFeOCAhromatogram ofa 0.5 ng/mL Standard 2xyjea Rc*e*rch < ?I{c2Sof43 Exygen Research : 684 : Page 167 of 217 MIN 313/023622 Exygen Study No.: 023-076 EhjtikiM atai Noe BlM-C23-a71 RcvMe l ligure 10. RCeopnrteasineinntgatPivFeHCShromatogram of S.Ong/ml. Standard Figureil. CRoepnrteasineinntgatPivFeOCShromatogram of 5D ng/mL Standard ExyjesBcsesrch Exygen Research : 685 : fifa 29Of43 Page 168 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxyjoaMethodNo: ExM023071Renuoo i figure 12. RCeopnrteasineinntgatFivFeOCAhromatogram ofa .t) ng/mL Standard Exygen Research : 686 : Page 169 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exyjco Method No: ExM-G23n71 Rrvixioa I Figure 14. RAenpalryezseendtfaotirvPeFCOhSromatogram of a Reagent Blank Sample Figure 15. ARenparlyezseendtfaotriv.PeFCOhAromatogram ofa Reagent Blank Sample Exygen Research : 687 : Pi* 31 of t3 Page 170 of217 MIN 313/023622 Exygen Study No.: 023-076 ExygenMethodNo;ExM-023-071Reviooc l Figure 16. RAenparlyezseendtfaotirvFeFCHhSiTimatogram ofa Ctotrol Liver Sample Figure 17. RAenparlyezseendtfaotirvPeFCOhSromatogram ofa Control Liver Sample Exygen Research : 688 : Pete 32e f Page 171 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exyiea MefeodNo: ExirMZWT] BevisiOQ1 Figure 18. ARenpalryezseedntfaotirvPeFCOhAromatogram of a CtmtroI liver Sample Figuris. RAenpalryezseedntfaotirvPeFCHhSromatogram ofa Control Serum Sample BroganResearch Exygen Research : 689 : P*i33oM3 Page 172 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxygesMethodNo: &SM-QZ3471Rcviijon 1 Figure 20. RAenparlyezseendtfaotirvPeFCOhSromatogram ofa Control Seram Sample Figure 21. ARenparlyezseendtfaotirvPeFCOhAromatogram ofa Control Serum Sample HxyjwRMMreh Exygen Research : 690 : Pi.jcJ4oi43 Page 173 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxypsMethodNo:EiM-023-071Steviiioa 1 figure 23. RAenparlyezseendtfaotirvPeFCOhAromatogram ofa Control Urine Sample Exygcatoccch Exygen Research : 691 : Fife 39Of43 Page 174 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exygca Method No: ExM-Q23-<J71 Revitioo 1 Figure 24. RFoerptrifeiseednatatti1v0enCgh/growmitahtoPgFrHamS ofa Control liver Sample Figure 25. FRoerptrifeiseednatatti1v0enCgh/growmitahtoFgFrOamS ofa Control liver Sample ExyjenResearch Exygen Research : 692 : Ptg&36of43 Page 175 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exype Stated Na ExM-07tR*ioo 1 Figure 26. RFoerptrifeiseednatatti1v0enCgh/growmitahtoPgFrOamA of a Control Liver Sample Figure 27. FRoerptrifeiseednatatti5v0enCgh/growmitahtoPgFrHamS ofa Control Liver Sample E xyjeaisw sds Exygen Research : 693 : Pace 37 of43 Page 176 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxyfaxMtfbodNo: E*M-023-07LRcrnsiaa l Figure28. FRoerptrifeiseednatatti5v0enCgh/growmithatoPgFrOamS ofa Cbntrol Liver Sample Figure 29. RFoerptrifeiseednatatt5iv0enCgh/growmitahtoPgFrOamA ofa Control LiverSample ExyfcaReward! Exygen Research : 694 : Page 3J of43 Page 177 of 217 MIN 313/023622 Exygen Study No.: 023-076 EzygecMethodNo:BxM-023-071 Revirioc 1 Figure 30. RFoerptriefiseednatatti1v0enCgh/mroLmwatiothgPraFmHSofa Centro! Serum Sample Figure 31. FRoerptrifeiseednatatti1v0enCg/hmroLmwaittohgPraFmOoSfa Control Serum Sample Exyjeaj Rcaeirch Exygen Research Pie39tf43 Page 178 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exyjso MethodNa E*M-Q23-07I Revkicn 1 Figure 32. RFoerptrifeiseednatatti1v0enCgh/mroLmvartlolhgPraFmOAof Control Senna Sample r . Figure 33. FRoerptrifeiseednatattSivOenCg/hmroLmwatiothgPraFmHoSfa Control Serum Sample Exygen Research : 696 : F its) of43 Page 179 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjeaMttbodNeeEaii-023-071 Rcvisioc I Figure 34. RFoerptriefiseednatatti5v0enCgh/mroLmwatiothgPraFmOoSfa C&ntrolSerum Sample Figure 35. FRoerptrifeiseednatatti5v0ecCg/hmroLmwatiothgPraFmOAofa Control Serum Sample r' BsygsaRtacccfa Exygen Research : 697 : PV4lQf Page 180 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exytcr. No: ExMCZM' Kiyuot 1 Figure 36. RFoerptrifeiseednatatti1v0enCg/hmroLmwatiothgFraFmOoSf Control Urine Sample r*- Figure 37. RFoerptriefiseednatatti1v0enCg/hmroLmwaittohgPraFmOoAfa Control Urine Sample ExygenResearch Exygen Research : 698 : Pifc42ci43 Page 181 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExygeaMe>odNo:ExM'C23-0r71 Rgvixioc 1 Figure 38. RFoerptrifeiseednatatti5v0enCgh/mrolm. watiothgPraFmOSofa ControlUrine Sample Figure 39. FRoerptrifeiseednatatti5v0enCg/hmroLmwatiothgPraFmOAaf a Control Urine Sample SxTjca Research Exygen Research : 699 : Pigc43of43 Page 182 of 217 MIN 313/023622 Exygen Study No.: 023-076 Analytical Method: (MPOeotShfFoP)deironfflRuAoanrtaoUloycrsGitinasCenf,/eoMSsrueStrlhfuoemnDy,elatFnelrdumoLriinivdaeetriobny (Exygen MethRoedviNsioo.nE2x)M-023-071A, Exygen Research : 700 : Page 183 of 217 MIN 313/023622 Exygcn Study No.: 023-076 TITLE , Method of Analysis for thRealDUerteinrme,iSneatniaornyoafnPderLfiiuvcerroboyctGanCe/sMulSfemyl Ffacodc (POSF) in AUTHORS John Flaherty and Alan Sensue DATE REVISED Angnat 30,2002 SPONSOR 3MBCraoird3piMonrgaCt2ee2Tn0t-oe2xrEic-o0l2ogy SL Paul, MN 55133-3220 PERFORMING LABORATORY St3a0tBe5x8CyoRgleleesageRea,recPehAtnDn1rhi6v8e0! METHOD NUMBER ExM-023-07LA, Revision 2 ' TOTAL NUMBER OF PAGES 34 Exygen Research : 701 : Page 184 of 217 MIN 313/023622 Exygen Study No.: 02SO76 Erjrtm MethodNo.ExlM2WT71ARtv.2 MANAGEMENTAPPROVAL Cto F/lahfte)rty / / / f LEaxbyogreantoRryesMeaarncahge* 7/ D^ate ExygenResearch Dale 2- ______ X f-JU L-D J JS3oMphonnCsLoo.rrpBRomreaplcrnebTseoenfcfthaPntiohvleDog,yOH, DABT Date Exygen Research : 702 : Pagc2 of34 Page 185 of 217 MIN 313/023622 Exygen Study No.: 023-076 Bxjjm MetedNo. BIM23-071A Be.-2 TABLE OFCONTENTS ' I MTAABNLAEGOEFMCEONNTTAEPNPTRSO--V--A--I----------------------------------------------------------------------------------------------------------------------------------------------------32 LIST OF TABLES-----------------------------------------------------------------------------------------4 LEST OF FIGURES_______________ :--------1--------------------------------------------'--------5 1. Z3. CESHUXEMPMEMRICAIMARLEYSN--AT--AN--LD---C-S-OU---MP-P-P-L-O-I-E-U-S-N----D-----S-------------*---------------------------------------------------------------------------------------------------------------------------------------------------------------------------J.87S 33332...1243.... CPESrtqhaemanpmdapraemar*dtei!*on*-nt_-a_-o-n_-f-_d-S_-St-_aa-_-np-_d-p_-al-_ir-c_d-a-_-S_---o-_-l_---u-_-t_-i-.o_.---_n--_s-_.-.---_.-._-..-_-----_----_-----_----_-----_----_----_-----_----_-----_----_-----_----_-----_----_-----_----_-----_-----_---_---_--.-_.-.-.-_-----_---_---_---_---_---_---_---_---_---_--_----_.888 34.1. 33.A4.33.. FSCotaorlctiibkfrisacotailtouiontinSo/tnaC.n..a.d.H.a~brmd..s.i..i..c...m........S.--..t.r..a..d-..-i.-n.-..-i.-.S-..-o.-.l.u..-t.i-.o-..-.n-..-s.--.~.----------------.-_-------------------------------,---------------------------------.-.-.1.330 4. METHOD__________________________ __ _________ _____ _______I;,________ 11 444...123... FSBkaamrtwcphDleSiePatrguorcpaem-s--s--i--n---g-----------------.---------------------------------------------------------------------------------------------------------------------------------------.-.----------------------------_--_---_---_---_---_---111111 4.4. 4.5. 44SQ..a44nm..a12mp.. lSleLiteAiirtviunoemarnlA.y-/-snU-i-asr-lP-iyn-rse-ei-psA---a--nr-ax----ti-iy-o-in-.i.-s*.---.-.._.--._.--._.._.~_.-.~-_.-.~_.-.---_.-._.-._.---._----_--_----__---_-_--__--_--_--__---_-_--_--_--__--_--_--__--_--_--_--__---__---_-_--_-_--_-_--__--_--_--__--_--_--_-_-_--_-_---__---_-_-,,__-.11112322 44..552. 2. A4.J5S.43.. CALanltiGobsrCaa/mtMiopnSleCSr uOyrivpteearnPartaionncgdedCOuoprneedrsai.t.t.ii.no..gn..sC......o.....n.....d.....i...t...o.....l...a...._...._...._...._...._...._...._.._...._......_..._..._...._...._..._...._...._....._..._...._..._.._. 111543 Sample Analyais.------------------------------- ----- .....---------- *----------------15 5. 444C...a678l...coPTAleiacnrrcfsieooeproRmtsa.ean_qn_cuc_eie_rCe_Cdr_irt_fie_otre_rir_aAi__an__.._a_..l__.y.__.s.__.i.__s...__....__....._....._......_...._...._...._.._.._.._.._.._.._.._.._.._.-_,____________i_n_,,_n_T_-_.._.._..__,,_.._-_,-11116777 _ . . 6. Safety-- ............... -- .............................- ,,,-- ____ _____i<? Exygea Research Exygen Research : 703 : Page3cf34 Page 186 of 217 MIN 313/023622 Exygen Study No.: 023-076 ZxnBlMcodNa. &cM-OS-071A.1t.2 UST OPTABLES Tibie I- Recovery Summary ofPOSF in Rt Serum ---- ...............-- .....------------ ----- 20 Table . Recovery Sram ary o fPOSF in KU U m --------------------------------------------21 Table HL Recovery Summary ofPOSF b K it Liver ,,--------------------------------- - -- 22 BxygcsRcscccii Exygen Research : 704 : P*ge4of34 Page 187 of217 MIN 313/023622 Exygen Study No.: 023-076 Ex?gcsU0&odNo.Bxi&Z3-O71AKjtv.2 LISTOFFIGURES .... . Figaro 1. Calibration carve for POSF ...................... -- ...............................................23 Figaro 2. Representative Chromatogram of1 0.10 jig Stodard Injection ofPOSF__ 24 Figaro 3. Representative Chromatogram ofa 0.01 pg Standard hgecdon ofP O S r__ 25 Figuro 4. Representative QhmTnatftgram of a Control Rat Serum Sample 26 Figure 6. Representative Chromatogram of a Control Rat Liver Sample------- --------- 28 Figure 7. Representative Chromatogram of a Fortified Rat Scrum Control Sample (0.1 pg hgecticn ofPOSF)-----------------------------------------29 Figure 8. RSaemprpesleen(0ta.1tivpegCInhjreocmtiaotnoogrfaPmOoSfFa)F--o--r-t-if..i.e..d...R...a..t..U...d..n...e...C...o.n..t..r.o..l......................30 Figure 9. RCeopnrteroselnStaamtivpeleC(h0.r1ompgatIongjreacmtioonfoafFPoOrtSifFie)d--R--a--t-L--i-v--e-r---------- -- ----- 31 Figure 10. RCeopnrteroselnStaamtivpeloC(h0r.0o1mpatgoIgnrajemctoiofnaoFfoPrOtifSieFd)R....a..t..S..e..n...a..n..... ......................... ,,..32 Figure 11. Representative Chromatogram of a Fortified Rat Urine Control Sample (0.01 pg Injection o f P O S F ) ....................................... -- 33 Figure 12. CRoenptrreoslenStaamtivpeleC(0h.r0o1mpagtoIgnrjaemctioofnaoFfoPrOtifSieFd)-R--a--t-L--i-v--e-r--------------------------- 34 Exygen Research : 705 : P***$of34 Page 188 of217 M IN 313/023622 Exygen Study No.: 023-076 ExyjeaMethodNo. ExM-023-071ARev. 2 1. SUMMARY TPGebCriG*/MocSrre.opoocrttanedseutaIiflc*nyla FlMuoertihdoed(PoOfSFA) nianlyRsiast Uforirne,thSeeruDmc,tnannmd altiivoenr boyf Analysis of headspace g u ii used to detect POSF in n t urine, terum, and liver. sQpueacntrtoifmiceattriyon(GoCf/MPSO)SaFnalyissisa. ccomplished by gas chromatography / mass The proposed Emit of qcxntitaiion (LOQ; the lowest fortification, specified by the mmeetthhoodd iws 2hi^chngg/imveLs (afodresqcurautem raencdovaenrnyo )acocro2r.d5inpgg /tgo(fEoPr A Hy cgtu) ifdoerliPnOesS)Ff.o r this 'Tp2cuehtrroivcsee1mn2i st.ertsehhcooodwvenWriiaenssFadiregevuesrhleoopw1e.ndRiunespTirnaegsberknasttaItu,irvIieIn,ceah, nrrodamtI IsaLetorugAmrar,meapsnraedrseernsethtaotliwivvneeric.nalFiTbiygraputiricoeansl Exygen Research : 706 : Page 6 of34 Page 189 of217 M IN 313/023622 Exygen Study No.: 023-076 Exyjta MethodNo. B3M-023-071A Rev. 2 2. . EXPERIMENTAL COMPOUNDS The troctmc forPOSF is givenbelow. PCOheSmTical Name: Perflcarooctanesolfenyl fhioridc (PO$F) CMCAhoeSlmec#iuc: la3al0rF7Wo-3rem5i-ga7hlat: CgFnSQzF 502 CF3(CF2)gCF2- S --F O Exygen Research : 707 : Pifie 7of34 Page 190 of 217 M IN 313/023622 Exygen Study No.: 023-076 ExyjeaMethodNo. B4M2MJ71ASIY.2 3. CHEMICALS AND SUPPLIES Cbdocals Methanol (MeOH) Grade HPLC ' Source EM Science Catalog No. MX047S-1 Standards Standard Lot Number FPleucoftroidoero(oPcOtaSaFca)ulfonyi 1S721HQ P orfty(% ) 98.6 SSoigumrcae 3-3. E quipment and Sctfuxs Equipment Balance, analytical (display at least 0.0001 g)~* 125-mL amber glass bottles Supplier ^KMiorottbletre^ 2IQ0-niinLLhaemadbsepragcleasv3ials* C.`jal-aysflsdaArejpjigpfertesqaunidpmTeonfat r(ogertrfi4cnfgl,fiarHirsr y lfnHrrn" (gspoeablc tabes etc.)______________________ G C ^ system LEAP Technologies Combi-PAL sutoeamplcx- Kimble ' vvaaTrreiioocuuhLssncsaoauclpoppgppilleiieserns Aa described in A ssseedccettaiioocnnri44b..cS5d..in Nwifoatshteed:yetEecraqmnuiibvneaeldeshntootwmbenat1teo2ripmarlLsod;muthacyeesbseaevtsiisauflbasscwttioteurryteerudessfeuodlrtfso. r*thAecmptu*eartnlhfhowedyfalvdiasnlpitdaiwcaetiomvno.wlutmyvef 3.4. Pripakation o p Standard Solutions Analytical standards are used for three purposes: 1. uCseadlibtoractiaobnbSratlaentdhaerrdess-pTonhseeseofsttahneddaertdescatorer upsreedpainretdheinanmaelythsaisn.ol and are BxygcsResearch P agtS oO * Exygen Research : 708 : Page 191 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExygtnMethodNo.ExM-02J-C71AR*r.2 2 . Laboratory Control Spikes - These- fortifications are prepared at dcoentecnenritnraetiaonnaslyctoicrarelsrpeocnodvienrgy.toLtahbeorLaOtoQry tcrounMtroxl LspOikQesaanrde aprrcepuasreedd tion control matrix. > 3. Matrix Spikes - These fortifications are prepared by spiking into tbe field ssaammpplleesmaatt4rixx LonOaQn.alMytiactarlixreacpoivkeeraya. re used to evaluate tbe effect of die Tcohrerect pfroorpteovrotilounms eosfoaof ituhtecsttoansodlavrednstmaracymboaivnatariineeddb.y the analyst as inng as the 3.4.J. Stocksolution Prepare an individual stock solutions of -2000 p.g/mL for POSF by weighing oat 200 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 100-mL .volumetric flask. The stock 2soCluttioon6*(Cinaan1d2i5s-amsLsiganmebdetbr gelmasasnbuoftatcletu) riesrt'os ebxepsitroatrieodn,indaater.efrigerator at /3 .4 .2 . F o rtific a tio n C a lib ra tio n S ta n d a rd S o lu tio n s a. PPcorOLeSpvFaorbeluyamafedotdrrtiiicnfifgcla-at2sick5GamnfcdLabloibrfirnfaegtkem-2ts0ot0avn0odplaugrmd/mesoLwlusitftoeiocmnkeosfoefah1nu0oi0oL0a gingtfoaoaL5f0b-r b. PS rrevrcl/Sratrfeoriftb,href remanrinc frrrfr'firjf?tin 2T5grvu^rsrtHofL &om 1s0ta0n0daHrdgtsaollutiaonnstoolf Example: Five zmeroliters of fee 50 pgfcriL solution spiked into 0.1 g of l2i.v5erpg(ofera1L0)0foprLtifoicfasteiornu.m or urine) is equivalent to a 2.5 ppm QL5 jig/g or Store all fortification standard solutions in a refrigerator (in 125-mL amber gdalatosspbreopttalreesd).at 2C to 6C for a maximum period o f one year from tbe Nmoatyeb: eAprdedpiatiroendailfcnoencceessnatrryat.ions of Fortification / Calibration Solutions Sxygco H o a r d : Page Sof34 Exygen Research : 709 : Page 192 of 217 MIN 313/023622 Exygen Study No.: 023-076 Extjci MethodNo. Esh.W23-07iA Hcv. 2 3 .4 J . C a lib ra tio n Sta n d a rd s ' GC/MS analysis calibration standard levels are determined by spiking a shaomperpaadr0los.l1ppaarigmcaeto.eruaEntmxtlaie(vmtaeynprpt)lie,ocsafeolvclyfoacpna5otlrrioabpltrLimant)giaotirntnihtxsoeta(1nsit0da0asnroddpiaLllreeddvf,iexvlasinraaadlrteiwensxhhjeaoimccwthinnacganosdn1fntoalxlilnnouLswrisnao:nef OtgtPeL/w)p 13212350305030005 VSpo&tanao* Wis5555 Vwoi11111tla22222.n)e IVnjoectatiro*n (n11lI1lL) (m) - 000500...0.00.321412310021 Exygen Research : 710 : Pigs 10cf34 Page 193 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxypgnMfl&odNo. ExM-Q23-071ARev.2 4. METHOD 4.1. FlowDiagram Tdehsecrifplatiwondoifeegancmh steopf. the m'ethod i* giver- below, fijDowed by t detailed Method Plow Diagram WFeoigrthif-y0s.1amgpoleftsidveesriognramteed.asa(ussremeala1it0mr0ixmp;seLpdiiokafetessleyar)nudm/o(rolramboinraet)oirnytocohnetarodlspspacikeevsial (heat to 60*HCefaotrt--o38m0*iCnuftoers~i4fumsiinn4gutaesLfEoAr mPaCncumalbiin-PjeActLioinnfmampler) Inject 1mL4headrace GC/MS4Analysis A 2. Samtue Processing Nbseeorfuosmraemoupsrelue.ripnreocseasmsipnlegsims unsetedbeedalfloorwseedrutomcoormuprlienteelysatmhapwlesto. rHooomwetevmerp, efrraotzuerne Fi(ncotoornKtaavine1ri2nsgammfoLpelelhisve,aerdresmspaomavcpeeleay)piaipnLrtooxaiImmfraemteeclzydera0ii.rf1ityigmrsaeematols a((nwcaaiplny)ksifssoteeilxlvcfeiraeoLdzsenoP)nlaeandcdeaypv.liaacles 43, BatchSetw a. A batch ofsamples ihccld not containmore than 20 field samples. b. Ebcolaancnchkenbutarstacitnhigooncf(otsynaptmriocplallemlysabaternitaxwl)yezeaennddthmeounLseOt Qminacatlnruiddxel OcaoxtnLlterOoaQsl tflooenrvteeiflsice)o.dntarot la(mknetohwodn Pige 11 of34 Exygen Research : 711 : Page 194 of 217 MIN 313/023622 Exygen Study No.: 023-076 Eiyxcalhftad No.Biia3-C71A Re.S c. kbAnatsoelwodnsutpcooonnnecthnfeitetsnlpdtoicnaasm.orp'Alsedreidnqitueiioarnecmih] ebnsatlmsc.hplae*mitnt itlhieobbietcthenptunyteailysofobretiffioerdtifiitedt d. Samples will be analyzed in duplicate. 4j4. Sammx Analysis PKETARATWPf 4 .4 .1 . L iver A nalysis a TReigmhotlvyec-a0p.t1hegvioafLliFvoerrtisfaymspalme ptoled(ipflnaecceesinsatroy)a v1i2a szynrLinhgeea. dspice vial b. Hmmearrtt<to-*8jf0T*iCinfgora~L4EmAPinCutoems fboi-rPmAaLnusuatloisaagmecpdleorn. or beat to 60*C for ~3 c&. IAnnjeaclyt z1emsaLmhpelaedssupsaicneg. GC/MS. 4.4J. Sinm/Urbu Analysis a. MTiegahstulyreca1p00thpeLvioaLf sFerourmtify(erammipnlee)(isfanmepcelessianrtyo)avi1a2smyrLinghee.adspace vial b. c. HIrrnnejreauctrTtt^o1 m8jf0Lr*wChmefaogdras~pL4aEcmeA.iPnuCteosmfboir-PmAaLnuaaultoinsajexcsptiloenr. or heat to 60*C for - 3 <L Analyze samples using GC/MS. SxygenXMm& Exygen Research : 712 : P*jb 22 of34 Page 195 of 217 MIN 313/023622 Exygen Study No.: 023-076 ErygtnMafcodNo.ExM-023-071ARev. 2 45 . QDAXTTTAnON 4S.1. CC/MSSystemandOperatingConditions t Instrument: HCehwrolmetta-tPoagcrakpahrd/mmooddcell56987930mSaeirsieasdGccastive detector HestekILTX-5012, 60mx0.25 rnmlD, M p n d f OvenTemperature: Hold at40Cfcr4 min^ ramp20C/imn.to I00C,hold fix 0 min. Transfer Line Teiiipcraim~ Ccrier Gu: GCH etd Pressure GCFtow Injection Mode Injection liner ibjcctico Purse Dcky Purge Flowto SplitVent Injection Volume Electron M ultiplier Voltage Scan Mode MonitoredIons DwellTime Integrator Rttnakm Times Totalrun time z tsrc 220-C Be&nm mT5rpmitLrwi,Tfromamrfo0pr.2p-58ff0mhfprtoaa.in,/maralyminti.pctao8l03TTp0ITapL/msi.mho, ltdo a6t03p0spi,sbi ofoldr 05 Varies with GC bead pressure and oven temperature Splitless Lrrrm gNt 2 min. 10 mTJm'm lm L From ATONE+ 200V SIM m/z'i 69(quantitationion); 131,219 (qualifierloos) 75 ms Hewlett-PackardCbemStation Software ~3.4 mincies ~7 mrmit-* BxygeaResearch Page 13of34 Exygen Research : 713 : Page 196 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exyp=MaS*sdNo.Ei2n>07lAS*v.2 /tuUnanpUrOptrntmgCoiuiitioiis StuffyPlm Ho. ExP*023^>75 Etygsa StudyNo.023-075 TngtnTm^yit* LEAPTechnologic*Combi-PAL sutrwamplrr - Injection/ Volume: Headrace 7lmL. Incoiamr/ AgilaSorTemp: 6CTC Incobator/ AgitatorTunc; 3 m . Iacnbator/AgitatorSpeed: 400 PPM-2 sec. on, 18sec.off Syringe Temp: 70C Syringe Fill Speed: 50 pL/aec. Syringe inject Speed; 750(iL/c. SyringePullup Delay: 1sec. Pro-injccticn Delay. 100 msec. Post-Injection Delay: 200 msec. ExygenResearch Exygen Research : 714 : Page 14 of34 Page 197 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exyjea Medod No. ExM-025-073AR tr. 2 4 J .3 . C a lib ra tio n C u rv e P ro ce d u re s a. Prepare calibration standards by spiking an appropriate volume of standard sO(clfneoaoamrvvllitreiunoebr8atul)iri0tn.onapo*jtntrseCAieocoai)pftnn,tfiatoet6mrocsrne0otda,*taanhn)Cn,tfesudroei(oavanetallhriltladeoyemvldsi(tiaiayhrsn(talrcrejcinieanxoiGgcpsgntipC(ned-1ecgt/iid0hMtosi)e0nfohrSdhneosp.ieeaamtLadmitoIed(fednfsfhouopeteaesrdaoamicrtn7leeao(o0gdhtwvu*e)aiCnseaafet)stLolr.etourEwdlm(f)AehavhipPfceaepohlnaCrrddoscosxto(mrapianamnapbttdcapaieua-irtnPorerdis(lxAn1yiaetL)monmfoasoaLfutruoept)reo0lpy'e.omr1)hsofmiipngtgrehpuhitratlreceeetashesxett b. aUrseegweneiegrahtteedd lfionreaerascthanadnaarldytceurbvyeslifnoeraqruraengtrietastsiioonn. ulsiinnegari/sxtawndeaigrdhtcinargveosf psnoreurfamatkiwmbaeafrrrredeo.affcrvAoaemnlniybutrcshaateilcoiabcnlarialbscttriuaaonltnaidotaisnortdanssntoadthnafadrftdaomrefdoacuycanclbidsbecrteeaoxstitcborlaecutdiacoeudnsrtvmauetsoi.sisnttiHgcnaooHlwtPoeeuxvtCceleirhee,crdtmhm2ae0atf%tytoiotboanelf the total number ofstandards injected. e. The correlation coefficient (R) for calibration curves generated must be asp0ta3pnr9do2apr5rdias(toeRr3fsotee0p.rs9e8lem5v)ua.snttIsbfeetcoatlfaiskbaermantpiotloecs results ashdojuusltd ibfnaesltlrreuoamunetasnliytdzeoepdthe.erasetioEnm, iatrsi,dththene A typical cairbrarion curve for POSF can be fbqndm Figure 1. 4 .5 .4 . Sam ple A n a ly sis a. tPshtraeenpsdaaarmredpslsaeomilnupttloieosanbhiyenatwodsefpiogaehceisnevgailae(fldoIr(fclanivpeepcree)dsos)arhryme,aedsapsspiukarecineagnsa(afmpopprrloseepvrruiiasmLte avAnodlfuteumrrienfooee)f vmcaoniaantnoltrsuoaiaslml,lypceolietnncrjd.efioctiirtnottinhoneejedfscoatei(mohneeG,ftafCeold/ieMq)uvSoifatolrs(1yasmctpeopLmnr)od. xiotiifmoIenafaetceduhlsy(ihsnteafgaontudeardar)drLfn,oErsmrAa(miPatMppplCero(,oaxrmteimcboi8a-v0Ptee*Alr0yyL)),, three narrates at 60*C (fee syringe is heated to 70*C). b. Standards corresponding to at least five or more concentration levels (starting with foe LOQ level orbelow) must be included m an analytical set, c. Asent efonltliorewseedt btfycaalfboartacthioonfsstaanmdpalredss. shHoouwldebveeri,najebctaetcdhf'tmthaey bneogtincnoinnsgisot foaf ExygenResearch Page 15of34 Exygen Research : 715 Page 198 of217 MEN 313/023622 Exygen Study No.: 023-076 B xnea MrtxxJNo. BxM-flZ3-071A.Rrr. 2 mcaolirberattriiomn ctuwrevnetmy assitmbpelgeesn(egrartiendd. ing spites sod dnplicates) until a new d. The concentration of each samplc/fortification/ccntrol i*~dctatmmod from the ruRneenescsrpeauostlnsteassdremys,cauyaarvmbesemi,dqabublaalrewnarctdiktsaeaottmeordpesuldeppieoantolpsieqe1*ue0oko%tfamodreuiaeatysrioedbsefeidtehuuaeescehcfduoaruivnnfeadlcbyioyntem.eepxaotcrTuhanhpsdeoalmarsettpiasolnpendo.sanersIdetf exceeds the calibration curve. e. FRoerctoifviceraiteiosnouretsciodveetrhieesseflailmlinitgs mwiatyhibne6a0ccteop1te4d0w%itahrespcoonnssoirdaeprepdroavcacle. ptable. Rbfreaottzweseneeranutm2>1aCQndtCouur6nintiCel msoaanmleyp&dleagsy. mpruisotrbtoe satnoarleydsifsr,ozaennd arat t-1li0veCr,mour srtefbreigsetroarteedd g. Freiteelndtisoanmtipmlees winillwbheicrehpnoortepdeaakssNaDre(ndoettedcetteedcteadt )d.ieSacmorprelesspoinn-dwinhgichanpaelayktes LarOe QdewteiclltebdeirtepthoertecdoatisaNpaQad(ninogt qunainltyifteiarbeltee)n. tion time that arc less than the 4.6. Accxptanoc Crjtexia The following criteria must be met to ensure the presence of POSF: 1. The instrument must have adequate scnritivily (s/n > 10) tt the LOQ level. 2. MbcleaeantshekoucdnobtnilltaatnhiknesssmtohuuersctaennooalftytcthoeenactatsinlnetPvmOelmSs Fgiiriaeotanlteeivsretflhosaugnnrded.aiteerLthOaQn,thaenLalOysQis. wIfilal 3. R14e0co%veorfitehseoirfkcnoonwtrnolvaslpuiekse.sIafnadcomnatrtorilxspsipkiekefosilshoouutlrdidbeethbeeatwcceeepnta6b0leloainumatsiltyisds,eias6ni0so-tw1hae4rr0r%Sanptisekhdeo.suhlodublde beevaplrueaptaerdedbyanthdeanaanlaylzyesdt.toAdrteytemrmatirniex ifpirkee 4. ofAforfonemcytaoclftiaarbilleribnacturiaamoltcnibouesnltarastotinaofdnnadtroiadwfrsdtvhtthffeodaucftnacoldobibjuterolacdttbieobedne.a, cesuxtacrvtliuesd.tiecdHalomowuuestvltieesrro,mtfoeaexytcobeteeadel xn2cu0lm%udbeoedrf 5. The correlation coefficient (R) for calibration curves generated rp^y* he 0.9925 (R20.985). If calibration results foil outride these limit*, then BxygeaResearch Page 16of34 Exygen Research : 716 : Page 199 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxygcnMethodNo. ExM-023-07lARev. 2 astpapardoaprrdiajtoeesttheeprselmevuasnt tbseettoikfmsamtoplea*djsuhsotuilmd btreurtneaencatlyopzeedra.tion, and the 4.7. Pzsjormancx CxrrzKZA Tthheecfoomllmowenincgemtweontcoriftaenriaalymsiuss.twbheenpeursfionrgmaend iansstarusmysetnemtatisounitsaebti-loitpyttheastt,hbaesfonroet been used for this method. First Criterion: PdreetepramreinaenidffionejeincsttraumGeCn/Mt hSassatdanedqauradtecsoernrseistpivointyd.inIgftthoisfcoreiteersitai,mcaantendotLbOeQmetto, Second Criterion: RLGccorOaeietnnQefefrai,ricoasiutneeepntiastotmocofasefltftiad,obneesrdaatemathirroimpdgnlshiencoesausfmttrifvoaicevnyoefnb(ooRcerer2at)nmhntraeooalfyratieznoaeactndloyl.nelteeacvesatennlt0dra.t9ooti8bo5tbnaeiflneoivrnaecfllolsiun,edeferaaodnrmareliyngatrteef.ososreibOoenanln,ocawewlyittsfhhiosieas. A X T W f fiiCOim>FOR Analysis iimOnnjnaeaetcprpitpixeorrsonspx*oii,knm2eca)0atwenfliiytleall2ktdaehksaoeausmarepspt.ploerTfos,2xh0ie1msGmaamtCaet/lpMryliex5Sshbtaholnauranorlskuy.,gsih1sfoolafebfsooaremastpoelrtey(pccroeonpntaatrrmoaltiinosgpni6pkreso,tacanenddduarrde1 5. CALCULATIONS a. UppaesraeakmEqeautreaertasi))ongue1snietnorgacteatdlhcbeuylafstoteaentHhdeaPradCmhoceuumnrvstteoatfio(PLnO/sxSoFfwtwfeoaiurgenhdpterd(oignrlapinmge/.marL,rebgarseesdsiocmn Equation 1: Analyte fbendfug/mL) CPtafcAreSal-opInetercept^ xDF n F fidcr by whichthe foal vohmaem s tfihftcd, if necesary. ExygenResearch Page 17of34 Exygen Research : 717 : Page 200 of217 MIN 313/023622 Exygen Study No.: 023-076 Eryjtn MethodNo.ExM-C23-071ARev.2 b. Eijuation 2 waa used to Igplate the amount of POSF found (in pg/mL or MS/g)Equation 2: Total Analyte Found (pg/mL or pg/g) Analyte Found (ug/mL) x StandardVolume Added (mLI SW (g) or SV (mL) SW - ample weight, SV * sample volume c. wEiqthuaktnioonw3n awmaouunstsedoftPoOcSalFcuplraitoertthoeexpterraccetinotnr.ecovery for samples fortified Equation 3: Recovery (% )Total Analyte Found (Augn/amlyLt*e^A-dAdendal(yptge/FmoLu*n)d in Sample (ug/mL*) x 100 - (or ng/g) FcSthooaarmrtrearpserlpepeosolneridstnisinnwtghghapinacunhfraoppleoyesLtaeekOss,rQeaftwrieeenlitdldiloebtneeacmrtteiemppdloeearttweifdniolaelwscbNoheriQcrrehes(pnpnoooortntqdepudienaagnaktssaifniNaaarblDeylete)d(.enrteoettcetndeteditoeanccttteitmdh)ee. acsygfto Rascaicn Exygen Research : 718 : Page It of34 Page 201 of217 MIN 313/023622 Exygen Study No.: 023-076 Ejcjjo M ated No. E*t)23-07IAJtrY. 2 6. SAFETY Hie analyst should read the material safety data sheets for all standards and hrleaaabngoderalninttosgrysbtceaofnosdtrsea,rdsjssafacjfntnydrgtrnleamaagsgeenst,thsai,nsidnmcglleuotdvhieonsdg.. woUrksiengtminivfaemaael hporoedcsauatniodnws ewarhinegn Exygen Research : 719 : Fags 1$ of34 Page 202 of217 MIN 313/023622 Exygen Study No.: 023-076 Bcypsi MaOodNo. ElM23-071A Kv. 2 Table L Recovery Summary ofPOSF in Rat Serum Summary ofRstSerum Rocovari Sample Sampio tnjocted Sample A rt Strf Injected SW Atm Trial Amount(pg) Counts Amount (go) Counts #1 0.1 803475 0.1 538407 #2 0.1 5064S2 . 0.1 538407 #1 0.01 48925 0.01 51030 % Reoovery 113 94.4 92.0 Exygen Research : 720 : P ip 20 of34 Page 203 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjecMethodNo.ExM-023-07IA Rev. 2 Table n . Recovery Summary ofPOSF In Rat Urine Summary of Rat Urine Recover!** SaTmriaplle nt #i Sample Injected Amount(ps) 00..11 0-01 SamCpoluenAtsrte 456183381800 48038 Astma oIunnjetc(tuepd) 00..11 0.01 SCWouAnrtes* 688822558B6C 51030 Rec%overy 7707..76 94.1 ExygcaResearch Exygen Research : 721 : P*fP21of34 Page 204 of 217 MIN 313/023622 Exygen Study No.: 023-076 Eiyien Mabod No. EXM4J23-971ABv. 2 Table HL Recovery Summary ofPOSF in Rat Liver Summary ofRatLtvarRaeovariaa SaTmriaplle o*eAm...m.por1au,.nItnntii(,a.p1cg-4r).ao SamCpoluenAtsrea. ASmtdolrulnatc(tpagd) SCtoduAnmts #12 00..11 750622585776 00..11 662288557768 #1 0.01 45864 041 51050 Rsc%ovary 817124 894 ExygcaRmcaicfa Exygen Research : 722 : Page 22 c f 34 Page 205 of217 MIN 313/023622 Exygen Study No.: 023-076 Bxy|*nMethodN.E*M-023-07lARrr.2 Figure 1. Calibration curve for POSF ' ' ' ExytenKecnrch Exygen Research : 723 : File Z3 of34 Page 206 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExygeaMctbod No. BxM-023-O71ARrv. 2 FSitganudrear2d. InRjeecptiroenseonftPatOivSeFCliromatogram of a 0.10 pg . s xa\fi23tfn\naE^\f?m209\in9m^ StmTqUiatSsInzS2isiaiadtndiewr>!i ^aSKpM0aoa,c tMwi ae*I--/*iSt*laigX*iaJ4a. a x fiaq s*4i S(es Uflb vial HxyjtnStaecdi Exygen Research 724 Ptfe24of34 Page 207 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxyjcnMethodNo. BxM-023-07LA.Rev.2 FSitgaunrdea3rd. InRjeecptiroenseonftaPtOivSeFChromatogram ofa 0.01 p.g vu* L i\m -m \n w n \rB W g i\ro o n ii TfacygmRr rarr>i Exygen Research : 725 : ofM Page 208 of 217 MIN 313/023622 Exygen Study No.: 023-076 BijpaMe6odNc.EM4S3-07!AJtar.2 yS(1jag0m0nprpleeL4.RatRSeeprruemse)ntative Chromatogram of * Control jg `\m-m\BMaa\t7m a u w a u 3 3 5 * .; 3 W | | f S i & S S t e S ? Exygen Research : 726 : Page 209 of217 MIN 313/023622 Exygen Study No.: 023-076 Bryj MttiudKo. BHM23-C71A te r. 2 SS{1alg0m0uprpeleLS.RatRUerpinrees)entative Chromatogram ofa Control i'IipinJ\-W9tCtw/X`-SnIo#IO\9MaO9wZ1nhnvt Owiaratlaa.ewW* *fM-c,gB1toMMttwTjlUt1o Bqiate g rcL Exygen Research 727 : P*fe 27 o f34 Page 210 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExyjenMethodNo. EiM-02M71ARev.J fS(0iag.m1ugprelRe6a.t Representative liver) Chromatogram of -- a Control n j g j .! J W 1 tiM I>m & i lo t K U m 3G-i*cl l *! CleonvtaTci**oa*iioi gRaweIi*irj.. t*. Utf- t u t Exygn&e*rch Exygen Research 728 : Page28 cf34 Page 211 of 217 MIN 313/023622 Exygen Study No.: 023-076 ExygmMiJodN<xExM-0Z3-07LAfcrT. 2 RFiagtuSreer7a.m CRoenptrreosleSnatamtipvleeC(0h.1ropmgaItnojgecratimonooffaPFOoSrtFif)ied Kla K_U__c _tf_cf_io_ iSMiMti9L.joouXc3i/ttma3eShOmpo/ut&taute*la v)Xa.,nm**i,a li t ft n 1nT1lAselt . 1 1 11 11 1 I I 1 '. k______ :______) ' \ ExygenR**e*rch Exygen Research : 729 P<029af34 Page 212 of 217 MIN 313/023622 Exygen Study No.: 023-076 Exjjcn M edial No. EiM-OZ3-<rrU. Itov. 3 FRiagtuUreri8n.e CRoneptrroelsSenatmatpivlee(C0.h1rpogmIantjoegcrtaiomnooffP*OFSorFti)fied i tiVm-m\xac>a\e7sioai\rau>T.s QB/U i t ExygaRocnxi Exygen Research Page 30 of34 Page 213 of 217 MIN 313/023622 Exygen Study No.: 023-076 EireaM c& ixlN o.2x14403-071A Xar.2 FRiagtuLreiv9e.r CRonetprroelsSenamtatpilvee(0C.1hr(oigmIantjoegcrtiaomn ooffPaOFSoFrt)ified ExygaoRcgcch Exygen Research : 731 : Pifo31of34 Page 214 of 217 MIN 313/023622 Exygen Study No.: 023-076 BxyfeaMethodNo.BaAtC23-C71ARev.2 FRiagtuSreer1u0m. CRoenptrreosleSnatamtipvleeC(0h.0ro1mfiagtIongjreacmtioonfoafFPoOrtSifFi)ed t feV8>*mYnuBXtt\e7ataaVroK2u.8 t r%xphrOQm,Oili/uM330*t0M*i3qfimha>.0ed.Ipiai P ri lu i. g R Mt&rthof xt, U IQ ExygenRacarcb Exygen Research Page 32 c f34 Page 215 of 217 MIN 313/023622 Exygen Study No.: 023-076 MKW No. Iter. 2 - SFiagtuUreri1nLe CRoenptrreoileSnatamtlpvleeC(0h.0r1ompgatIongjencutnioonfoafFPoOrStiFfi)ed 1'i^&3*a\eea/m-md\nnmia\M.07***MIM3la**yr"oa2pi7m.&mt tygcaR anrch Exygen Research : 733 : Pit 33 of34 Page 216 of217 MIN 313/023622 Exygen Study No.: 023-076 ExyjenMethodNo. Bj4-O25-071ARbv.2 RFiagtuLreiv1e1r CRoenptrreosleSnatmatpivlee (CQh.0r1ompgatIongjercatmioonfoafFPoOrStiFfi)ed i iu\e-e7i\*X3E3m\87OTM *Vmeau.B i*.t Sl*AaL<o,ix/i1mMaMS.9Mia/MMObiQ<MUa a.y-1aTMiakSi.i,a^gkttda. t 1lS*i Lwv m ial Exyjea Research Exygen Research : 734 : Page 34 of 34 Page 217 of217 MIN 313/023622 HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2001 THE DEPARTMOEFNTTHOEFUHNEIATELDTHKOINFGTDHOEMGOVERNMENT GOODLABORATORYPRACTICE IN ACCOSRTDAATNEMCEENWTITOHFDCIORMECPLTIIAVNEC8E8/320EEC LABORATORY TEST TYPE Huntingdon Life Sdeneas HWuonotlienygRdoonadResearch Centre AHr^lurcnnohtnijnLbgudryon PE284HS Analytical Chemistry Oiaical Chemistry EEEcnnovvsiiyrroostnnemmmeesnnttaall FTaotxeicity PTfoaxyisc/oClohgemy Testing DATE OF INSPECTION 154 January 2001 AwasgecnaerrrailedinosuptecatitontheforabcoovmepllaiabnocraetowryithasthpearPtrionfcipUlKesGoLfPGCooodmpLlaiabnocreatoPrryogPraramcmticee. Athtetvbaelitdimityeooffntobn?-cinlisnpieccatliosntudnioesdpeveriafotiromnesdwaetrtehefoseunfdacoilfitiseusf.ficient magnitude to affect * /th DHre.aRd.oUgeKr GG.LAPleMxaonnditeorring Authority : 735 : MIN 313/023622 HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2003 N-i- *i E THE DEPARTMENT OF HEALTH OFTHE GOVERNMENT OF THE UNITED KINGDOM GOOD LABORATORY PRACTICE STATEMENT OF COMPLIANCE IN ACCORDANCE W ITH DIRECTIVE 88/320 EEC LABORATORY TEST TYPE HWHuuonnottliilnneggyddRooonnaRLdiefseeSacrciehnCceesntre AHCPEalucm2no8tnbi4nbsHg.udrSyon AEEETCPhocnnlnioxyvvansisiilicyr/rycCooostalitnnhloecmmeCmgamlyheesCnneTmtthaaeellismstTFstrioaystxtericyity DATE OF INSPECTION 07** April 2003 AwagsecnaerrrailedinospuetcatitontheforabcoovmepllaiabnocraetowryithasthpearPtrionfciUplKes GoLfPGCooodmpLlaiabnocreatoPrryogPrraamcmticee. Athtetvhaelitdimityeooffntohne-cinlisnpiceac!tiostnudnioesdpeevrifaotriomnesdwaetrtehefsoeunfadcioliftiseus.fficient magnitude to affect C* `P3 - Dr. Roger G. Alexander Head, UK GLP Monitoring Authority 736 : MIN 313/023622 HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2005 Department of Health THE DEPARTMENT OF HEALTH OF THE GOVERNMENT OF THE UNITED KINGDOM GOOD LABORATORY PRACTICE STATEMENT OF COMPLIANCE INACCORDANCE WITH DIRECTIVE 2004/9/EC LABORATORY WAHHCPEaluuocm2nnoo8ttlnbiilnn4ebriggyHudddrSRgyooeonnsahLRdiirefeseeSacrciehncCeesntre TEST TYPE EEEATPChconnlnioxyvvansisiilicyrry/coCoostalitnnhlocemmCeamgmlyheesCnneTmthtaaeelilsmstFTtirinaosygtxteriycity DATE OF INSPECTION 7thMarch 2005 A general inspection for compliance with the Principles of Good Laboratory Practice was carried out at the above laboratory as part ofthe UK GLP Compliance Programme. At the time of inspection no deviations were found of sufficient magnitude to affect the validity of non-clinical studies performed at these facilities. Mr. Bryan J. Wright Head, UK GLP Monitoring Authority : 737 : MIN 313/023622 EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2001 THE DEPARTMENT OF HEALTH OF THE GOVERNMENT OF THE UNITED KINGDOM GOOD LABORATORY PRACTICE IN ACCOSRTDAATNEMCEENWTITOHFDCIORMECPLTIIAVNEC8E8/320EEC LABORATORY TEST TYPE HEESIPnyyu2feenf3otRi7lnekPgsXedaornchLCifeenStcrieences ACEEETMPcohnnlnuioxyvvantsiliiiaicyy/rrcgCooostaeiltnnlhcoenmmCeamgimclyheesiCenntyTmhttaaeeillsmsTFttirinaosytgxteriycity DATE OF INSPECTION 29* January 2001 Awasgecnaerrrailedinospuetcatitonthefoarbcoovme pllaibanocraetowryithasthpeaPrtrionfciUplKesGoLfPGCooodmpLlaiabnocreatoPrryogPraramcmticee. Athtetvhaelitdimityeooffntohne-cinlisnpieccatliosntudnioesdpeveriafotiromnesdwaetrtehefoseunfdacoilfitiseusf.ficient magnitude to affect DHre.aRd,oUgeKr GG.LAPleMxoannditeorring Authority : 738 : MIN 313/023622 EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2003 THE DEPARTMOEFNTTHOEFUHNEIATLETDHKOINFGTDHOEMGOVERNMENT GOOD LABORATORY PRACTICE STATEMENT OF COMPLIANCE IN ACCOKUANCfrW TTHDntECnVF. 8 3 E E C LABORATORY TEST TYPE EH>tEWS~/Vyyucn-2eeft-up3f-Rotim7loekFs^XedaorochLiCfeenStcrieences EETAEMPhcannnuexyvvftesfdiiayorr/ygCoosdtmtnnthleoommeimtjemiyefeCtnsvTthtaaemilaTtFsitadatxertydty 22D--ATAEpOrilF2I0N0S3PECTION Awagsecnaerrrailedinospuetcatitonthetoarbcoovme pllaibanocraetowryithasthpearPt rionfciUpiKes GoLf PGCooodmpLlaiabnocraetoPrryogPrraamcmticee. Athtetvhaolitdimityeooffntohne-icnismpieccstliostnudnioesdpeevrifaotiromnesdwatertheefsoeunhddVoiiftisourfficient magnitude to affect Or, RogerG. Alexander Head, UK GLP Monitoring Authority : 739 : MIN 313/023622 EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2005 Department of Health THE DEPARTMENT OF HEALTH OF THE GOVERNMENT OF THE UNITED KINGDOM GOOD LABORATORY PRACTICE STATEMENT OF COMPLIANCE INACCORDANCE WITH DIRECTIVE 2004/9/EC LABORATORY \ HSIEOEPuyycu2feecnf3ooRtli7ldnekPgsXedaornchLiCfeenStcrieences TEST TYPE AEEMTPCEhocnnlnuiyoxvvantsisliiaicy/yrrcgCooostaelitnnhlocenmmeCgamimclyheesiCtnneyTmtthaaeellismstFTtirinaosygtxteriycity DATE OF INSPECTION 12thApril 2005 A general inspection for compliance with the Principles of Good Laboratory Practice was carried out at the above laboratory as part of die UK GLP Compliance Programme. At the time of inspection no deviations were found of sufficient magnitude to affect the validity of non-clinica! studies performed at these facilities. Mr. Bryan J. Wright I M O * Head, UK GLP Monitoring Authority : 740 :