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CHEMICAL MANUFACTURERS

# AssooF'tz-o3'7·/.:l

# ?9

March 18, 1997

Dr. LynnGoldman AssistantAdministrator OfficiO' ofPrevention,Pesticides and Toxic Substances TS-7101 EnvironmentalProtectionAgency 401 

### :.1**Sttet~,** 

### SW, Room 637, EastTower Washington, DC 20460 DearOr. Guldman: TI:e Chemical Manufacturers Association makes available to thepublic and approp1iate governmentagencies final reports ofenvironmental,healthand safety**resea~** 

### that it manages. In

### keeping withthis policy, the following recently completedreportis :: enclosed: A 28-Day Repeated Dose Oral Toxicity Study ofHBCD in Rats Onlyvolume 1of3 is enclosed withthis letter. Volumes 2 and 3contains 658 pages andonlycontainrawdata. This reportdoesnotinclude**confid~ntial** 

### information. If

### youhave any questions, pleasecallme at 703-741-5637.

Enclosure Sincerely,

# Jfa./rn~~**~** HasmukhShah, Ph.D. Director, CHF
## 84970000012 

# Contains No 

# Cll

### FINAL REPORT VOLUME 

### 1 OF 

### 3 (Text and Summary Tables) STUDY 

### TITLE A**~f.-DAY** REPEATED 

### DOSE ORAL TOXICITY STIJDY OF HBCD IN RATS DATA REOUIIUMENT OECD Guideiine, Section 407 STUDY DIRECTOR Christopher P. Chengelis, Ph.D., D.A.RT. STI.TDY INITIATED ON Aprii 1 (. 1996 

### STUDY COMPLETION lJATE February 13, 1997 PERFORMITNGLABORATORY WIL Research Laboratories, Inc. i

### 407 George Road Ashland, Ohio 44805-9281 LABORATORY STUDY NUMBER Wl:L-186004 SPONSOR PROJECT NUMBER BFRIP 

### 2.0-WIL HBCD

Chemical Manufacturers Association Brominated Flame Retardant Industry Panel 1300 Wilson Blvd. 

### Arlington, V 

### sinia 22209 Page 1 of 925 WiL RESEARCH LABORATORIES, INC. ASubsidiary of Great !_akes Chem:cai Corporation, Ashland, OH 44805-!:!281 (419) 289-8700

# •

• wn..-186004 CMABFRIP


### A 28-Day Repeated 

### Dose Oral 

### Ioxjcity 

### Study of

## HBCP in Rats COMPLIANCE STATEMENT 

# ,. 

# •

Ibis study, designated 

### \Vn.-186004, was conducted in compliance with the United States

### Environmental Protection

### Agency (EPA) 

### Good

### Laboratory PracticeRegulations (40 CFR Put 

### 792), October 

### 16, 1989, the Organization for Economic Cooperation anti 

### Development (OBCD) Principles of 

### Good 

### Laboratory Practice [C (81) 30 (Fi.W)], the 

### Standud Operating 

### Procedures of

# wn. Research Laboratories, Inc., and the protocol as approved by 

### the sponsor (Chemical Manufacturers Association Brominatcd Flame Retardant 

### Industry Panel). 

### 1be protocol was designed and the study was conducted in accordance with 

### the OBCD Guidelines for testing of Chemicals, Health Effects Test Guidelines, Section 407, adopted May 

### 12, 1981. Study**~..rector:**

### 136~ Date

### WJL-186004 CM.ABFR1P 

# • A 28-Day Repeated 

### Dose Oral 

### Toxicity Stwfy 

### ofHBCILiJ.LRIU TABLE OF f:ONTENTS VOLUME I**~** 

### I. 

### Summary 14 

## n. Objective 18 

## m. Study Design 19 IV. 

### Experimental Procedures 

### 20 A. 

### Introduction 

### 20 B. Test and Control Articles 

### 20 1. 

### T.'!St Article Identification 20 

### 2. Control Article Identification 21 3. 

### Preparation 21 4. 

### Administration 

### 21 5. Sampling and Analyses 

### 22 

# • c. Animal Receipt and Acclimation/Pretest Period 

### 23 

### D. Animal Housing 

### 23 E. Diet, 

### Drinking Water and Maintenance 24 

### F. Environmental Conditions 24 

### G. Assignment ofAnimals to 

### Treatment Groups 

### 24 

### H. Clinical Observations and Survival 

### 25
T 

### Body Weights 

### 25 

## .... J. 

### Food Consumption 

### 26 K. 

### Functional Observational Battery 

### 26 1. 

### Home Cage Observations 

### 26 2. Handling Observations 

### 26 3. 

### Open Field Observations 

### 27 4. 

### Sensory Observations 

### 27 5. Neuromuscular Observations 

### 27 

### 6. Physiological Observations 

### 27 

# • L. 

### Locomotor Activity 

### 27 -3-' 

# . - 

### . ' 

### .,. ' '**~** 

### - ', ' '.;.

. . ' .. . .._ '


### WIL-!M004**~~~~ ~...,.**  ..**.&"'~** 

# • VO"**~VME** 

### 1 (eolltiaued) 

## fiG M. 

### Clinical Pathology 

### 28 1. 

### Hematology 

### 28 2. Serum Cbemistry 

### 29 N. 

### Pathology 

### 29 l. 

### Macroscopic BYaminalion 

### 29 2. Organ Weights 

### 30 3. Microscopic Examination 

### 30 0. 

### Statistical Methods 31 

### P. 

### Data Retention 

### 31 

# v. Results 

### 32 

## A. Clinical Observations and 

### Survival 

### 32 

## B. Body Weights 

### 32 

# c. Food Consumption 

### 32 

## D. Functional Observational 

### Battery 

### 33 

# • 1. 

### Home Cage 

### Observations 

### 33 2. Handling ObseiVations 

### 33 3. Open 

### Field Observations 

### 33 4. Sensory Observations 34 

### 5. Neuromuscular ObseiVations 34 

### 6. Physiological Observations 34 

## B. Loc:omotor Activity 

### 35 F. Clinical Pathology 

### 35 1. 

### Hematology 

### 3S 2. Serum Chemistry 

### 35 

## G. Pathology 

### 37 1. 

### Macroscopic Examination 

### 37 2. Organ Weights 

### 37 3. Microscopic Examination 

### 38 VI
. Discussion and 

### Conclusions 

### 39 

# •
-4-,.._ •**~** t**~** ' • a ". : • -/ • • r ' 1 ,-1 •**~** -

.. /**~** 

### .. ..**~** 

### ..**~** 

# .
\\"'L-186004 CMABFRIP



### • VOLUME 1 (continued)



# •

• vn. Key Personnel and Report 

### Submission 

# vm. Pathologist 

### Summary .:;wement

IX. Quality 

### Assurance Unit Statement X. 

### References



### -5- 41 

### 42 45

# •

•

• WIL-186004 CMABFRIP


### A 28-Day R!pated 

### Dose Oral ToxicitY 

### Study of

### HBCP in Rats 

### INDEX 

### OF TABLES 

### VOLUME 1 (continued) 1. Summary ofSurvival 

### and Dispositico 2. Summary ofClinical Findings: Total Occunence/No. ofAnimals (Priorto 

### Scheduled Necropsy/Dispositions) 3. Summary ofClinical Findings: (At Tune ofDosing) Total Occurrence/No. ofAnimals
4. Summary ofClinical Findings: (1-HourPost-Dosing) Total Occurrence/No. ofAnimals 

### 5. Summary ofClinical Findings: Total Occurrence/No. ofAnimals (Recovery Period 

### Observations) 6 . 

### Body Weights (Grams)

- Summary of

### Means 7. Body Weight Gains ·(Grams)

- Summary of

### Means 8. Weekly Food Cocsumption**(Gram~Animal/Day)-** 

### Summary 

### cfMean& 9. Functional Observational Battery Summary Incidence

- Home Cage 

### Observations (Week -1 

### Pretest Evaluation) iU. Functionai Observational Battery Summary Incidence

- Home Cage Observations (%] 

### (Week -1 Pl'etest Evaluation) 11. Functional Observational Battery Summary Incidence

- Home Cage 

### Observations (Week 3 Evalllation) 12. Functional Observational Battery Summary Incidence

- Home Cage 

### Observations [%] 

### (Week 3 Evaluation) 13. Functional Observational Battery Summary Incidence

- Home Cage 

### Observations (Week

### 5 Recovery Evaluation) 14. Functional Observational Battery Summary**IncidC"~ce** 

### - Home Cage Observations 

### [%] (Week 

### 5 Recovery EvaluatiOI·

-6-**~** 

### 47
49
51
53 

### 55 51 61 55
69
71
73 

### 15
77
79

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# - WlL-186004 CMABFRIP 

# • VOLUME l (continued) 

### 15. Yiunctional Observational Battery Summary Incidence

- Handling Observations (Week -1 Pretest Evaluation) 81 16. Functional Observational Battery Summary illcidence

- Handling Observations [%](Week -1 

### Pretest Evaluation) 87 17. Functional Observatlonal Battery Summary Incidence

- Handling Observations (Week 3 Evaluation) 93 18. Functional Observational Battery Summary Incidtmce

- Handling Observatiocs [%] (Week 3 Evaluation) 99 19. Functional Observational Battery Summary Incidence

- Handling Observations (Week 

### 5 Recovery Evaluation) 105 20. Functional Observational Battery Summary Incidence

- Handling Observations [%] (Week 

### 5 Recovery Evaluation) 111 21. Functional Observatioi'al**~.ery** Summary Incidence

- Open Field Observatioas (W (;ek -1 PretP..st Evaluation) 117 

# • 22. FunctioLal Observational Battery Summary Incidence

-**~n** Field Observations [%] (Week -1 Pretest Evaluation) 121 23. Functional Observational Battery Summ.u)' Incidence

- Open Field Observations (Week 3 Evaluation) 125 

### 24. 

### F!!!!ctiana! Obser--.::ticna! 

### .Bar..er-; SumiJ.Jar.**llic.~** 

### 1ecce

- Open Field Observations [%] (Week: Evaluation) 129 

### 25. Functional Observational Battery Summary Incidence

- Open Field**Observation~** (Week 

### 5 Recoven· Evaluation) 133 26. Functional Observational Battery Summary Incidence

- Open Field Observations [%] (Week 

### 5 Recovery Evaluation) 137 27. Functional Observational 

### Battery Summary Incidence

- Sensory Observations (\Veek -1 

### PrP..-test Evaluation) 141 28. Functional Observational Battery Summary Incidenc!!- Sensory Observations [%] (Week -1 Pretest Evaluation) 145 

# •
-7-**~~ ~~~~~---~** 
............UV\Ar"W' 

### CMABFRIP 

# • VOLUME 1 (coatiaued) 

### 29. FullctiOIIII 

### Observational Battery Summary Incidence

- Sensory Observati<D (Week 3 Evaluation) 149 30. Functioaal Observational Battery Summary Incidence

- Sensory ObseiVations [%](Week 3 Evaluation) 1S3 31. Functi'JDal ObseJVational Battery Summary Incidence

- Sensory ObseJVations (WeekS Recovery Evaluation) 1S7 32. Functional ObseJVational Battery Summary Incidence

- Sensory ObseJVations [%] (Week 

### S Recovery Evaluation) 161 33. Functirnal ObseJVational Battery Summary Incic:leoce

- Neuromuscular Observations (Week -1 Pretest 

### Evaluation) 16S 

### 34. Functional Observational Battery Summary Incidence

- Neuromuscular ObseJVations t.%] {Week -1 Pretest Evaluatio11) 167 3S. Functional ObseiVational Battery Summary Incidence

- Neuromuscular Obselvations (Week 

### 3 Evaluation) 169 

# • 36. Functional ObseJVational Battery Summary Incidence

- Neuromuscular ObseJVations [II] (Week 3 Evaluation) 171 37. Functional ObseJVational Battery Summary Incidence

- Neuromuscular Observations (WeekS Recovery Evaluation) 173 38. Functional ObseJVational Battery Summary Incidence

- Nfllli'QIIlnscular**~QM** 

### [%]**(W~** 

### S**~v~** Rv2lwation) 

### !75 39. Functional Observational Battery Summary Incidence

- Physiological ObseJVations (Week -1 

### Pretest Evaluation) 

### 177 40. 

### Functional Obserrational Battery Summary Incidence

- Physiological ObseJVations (Week 3 Evaluation) 179 

### 41. Functional Observational Battery Summary Incidence

- Physiological ObseJVations (Wee..1c 

### S Recovery Evaluation) 181 42. 

### Group Mean Motor Activity Counts (Week 

### -1 Pretest Evaluation) 183 43
. 

### uroup Mean Motor Activity Counts (Week 3 Evaluation) 187 

# • -8-wn.-186004 CMABFRIP 

# • VOLUME 1 (continued) 

# .ew 44. Group Meail Motor A<..1ivity Count'> (•.Veek 

### 5 Recovery Evaluation) 191 45. Hematology Values

- Summary ofMero•1 195 46. Leukocyte Differenili!l Count (%)

- Summary ofMeans 

### 203 47. Leukocyte Counts

- Summary ofMeans 

### 207 48. Serum Chemistry Values

- Summary ofMeans 

### 211 49. Gross Necropsy ObservatiClns Incidence Summary (Week 4 Primary Necropsy} 

### 223 50. Gross Necropsy**Observa~ons** Incidence Summary 

## 0N eelr 6 Recovery Necropsy) 224 51. Organ Weights (Gran&s)- Summary ofMeans (Week 4 Primary 1\
..:ropsy) 

### 225 52. Organ Weights (tirams) -Summary ofMeans 

# • (Week 6 Recovery Necropsy) 229 

### 53. Organ Weights Reh>tive to FinalBody Weights (Grams/100**G~ams)** - Summary ofMeans (Week 4 Primary Necropsy) 

### 233 54. Organ Weights Relative to Final Body Weights (Grams/**~00** Grams)

- Summary ofMeans (Week 6 Recovery Necropsy) 237 

### 55. ,**T~stomorphological** Diagnosis

- Summary Incidence

,Week 4 Primary Necropsy) 

### 241 )6. Histomorphological Diagnosis

- Summar} Incidence (Week 

### 6 Reco.rery Necropsy) 

### 255 VOUJME2 57. Individual Survival and Disposition 

### 269 58. Individual Clinical Observat:ons (Priorto Scheduled Necropsy/Dispositions) 273 

# , 59
. Individual Clinical Ot_:ervations (At Time ofDosing) 

### 279 

# • -9-

## . '/'**--~·-** 

## J;**·~·:·'** 

## :._:_...__ :.-.. :". -"' ,, (

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...**~·:·-;** 

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...**~** 1'

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rrrr 

### WD..-186004 CMABFRIP 

# • VOLUME 2 (coatinued)**fit&.~** 

### 60. Individual Clinical Observations (1-Hour Post-Dosing) 283 61. Individual Clinical Observations (Recovery Period 

### Observations) 287 62. Individual Body Weights (Grams) 288 63. Individual Body Weight Gains (Grams) 296 64. 

### Individual Food Consumption (Grams/ Animal/Day) 304 

### 65. Individual Functiooal Observational Battery Data

- Home Cage 

### Observations (Week -1 Pretest Evaluation) 312 66. lndividua' :-'· nctional Observational Battery Data

- Homer"' ·)bservations (Week 3 Evaluation) 320 67. Individual Functional Observational Battery 

### Data

- Home Cage 

### Observations (Week 

### 5 Recovery Evaluation) 328 68. Individual Functional Observational Battery Data

- 

# • Handling Observations (Week -1 Pretest Evaluation)**':\?~** • JL 

### 69. lndividua.l Functional Observational Battery Data

- Handling Ocservations (Week 3 Evaluation) 356 70. Individual FuncHonal Observational Battery 

### Data

- Handling Observations (Week 

### 5 Recovery Evaluation) 380 71. Individual Functional Observational Battery 

### Data

- Open Field Observations (Week -1 Pretest 

### Evaluation) 392 72. Individual Functional Observational Battery 

### Data

- Open Field Observations (Week 3 Evaluation) 408 73. Individual Functional Observational Battery Data

- 

## Oven Field Observations (Week 

### 5 Recovery Evaluation) 416 74. Individual Functional Observational Battery Data

- Sensory Observations (Week -1 Pretest 

### Evaluation) 424 75. Individual Functional Observational Battery Data

- Sensory Observations (Week 3 Evaluation) 440 

# • -10-

# ... wn.-186004 CMABFRIP 

# • VOLUME 2 

### (contiuued) 76. Individual Functional Observational Battery Data

- Sensory Observations (Week 5 Recovery Evaluation) 

### 448 77. Individual Functional Observational Battery Data

- Neuromuscular Observations (Week -1 Pretest Evaluation) 456 78. Individual Fucctional Observational Battery Data

- Neuromuscular Observations (Week 3 Evaluation) 472 79. Individual Functional Observational Battery Data

- Neuromuscular Ob[..!tvations (Week 5 Recovery Evaluation) 488

Pn: T**~..:·~-.,** •**~·** ·i:J<.**~ ~·**  ··· "''at;onal Battery Data

- . --.u**-~**

.......**~ ~** **lro..~** vCSe.vaL. ·; {Week -1 Pretest Evaluation) 496 

### - - )**~** 

### !'1. ludividual Functional Observational Battery DataPLysiological Observations (Week 3 Evaluation) 504 32. Individual Functional Observational Battery Data

- Physiological Observations (Week 5 Recovery Evaluation) 512 

# • 83. Individual Motor Activity Counts (Week -1 Pretest Evaluation) 516 

### 84. Individual Motor Activity Counts (Week 3 Evaluation) 532 85. Individual Motor Activity Counts (Week 5 Recovery Evaluation) 548 

### VOLUME3 86. Individual Hematology Values (Week 4 Primary Necropsy) 557 87. Individual Hematology V3.lues (Week 6 Recovery Necropsy) 565 88. Individual Leukocyte Differenticl Count(%) (Week 4 

### Primary Necropsy) 

### 569 89. Individual Leukocyte Differential Count (%) (Week 6 Recovery Necropsy) 577 

### 90. Individual Leukocyte Counts (Week 4 Primary Necropsy) 581 91. Icdividual Leukocyte Counts (Week 6 Recovery Necropsy) 589 

# • -11-

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### wn.-186004 CMABFRIP 

# • VOLUME 3 

### (coatinued)**~** 

### 92. Individual Serum Chemistry Values (Week 4 Primary Necropsy) 593 93. Individual Serum Chemistry Values (Week 6 Recovery Necropsy) 609 94. Individual Gross and Microscopic Description ofOrgans (Week 4 

### Prim2ry Necropsy) 617 

### 95. Individual Gross and Microscopic Description of**Organ~** (Week 6 Recovery Necropsy) 669 96. Individual C"'"p'l Weights and Final Body Weights (Grams) (Week 4 

### Primary Necropsy) 697 97. Individual Organ Weights and Final Body Weights (Grams) (Week 6 Recovery Necropsy) 705 98. Individual Organ Weights Relative to Final Body Weights (Grams/100 Grams) (Week 4 Primary Necropsy) 709 

### 99

. Individual Organ Weights Relative to Final Body Weights 

# • (Grams/100 Grams) (Week 6 Recovery Necropsy) 717



# • -12-

# •

• l!l"'rmo· N&lliM**···:Ii·l~~** CMA B-<'RIP
ft.**:.!~-fr.n** Repe?red Dv.5-e Oral**T!?~~:i~tdy** of**Tio~:::.;v** 

### i'l &w

A. Analyses of Dosing Prepa.rntions {Sponso7'-Ge1le1"3.ted Data) 

### B. Gravim<-.trk Analysis 

### ofT'tO.>iDg ?nrmulations (Wf'- P..ese:l!"ch I.abo:atories, Inc.) C. PrettS1 Climcal Cibs 1ativn'> 

### D. Scoring Criteriz for Ft nctional**Oos;~rvational** Battery E. Summarieo> of 

### V:tLdat'.:m**S~dies** 

### (WIL-9S;J26, WJL-99C1?..



### G . H. 

### I.
J . Wll.-99G34 <'- ' WIL·1fC")35)

WIL Motor Activity**Ifu~**orical Control Data Clinical Pathology Historico.i C**~nt·::>l** Data (WIL Research Labora10ries, IGc..) Study Protocol

-13- 

## ----:: . -

721**~29** 763 

### 766


### 782 813 818 

### 873

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# •

•

• WJL-186004 CMABFRIP



### I. 

## SJJMMAIY A 28-Day 

### Repeated Dolle Oral 

### Toxicity 

### Study of

## UBCD in 

### Rats

1betest article, hexabromocyclododecane (HBCD), 

### Lot #357/, in the vehicle, com oil, was administered orally 

### by gavage to 

### three groups ofSprague-Dawley Crl: CD BP rats 

### for a period of28 conset:utive days. Dose levels were 125 (low), 350 (mid), 

### or 1000 (high) mg/kglday, administered in a dosage volume of5 

### ml/kg. 

### The test

### groups consisted of6 males and 

### 6 females in the 125 and 350 mglkg/day groups and 

### 12 males and 12 females in the 1000 mglkg/day group. A concurrent control group 

### comprised of 12 males and 

### 12 females received the vehicle, com oil, for 28 consecutive days at a dosage volume ofSmllkg. 

### At the end ofthe dosing p'!riCid, 

### 6 animals/sex/group were eutbanized and necropsied. The remaining 6 animals/sex in the 

### control and 

### 1000 mg/kglday groups remained on-test untreated for a 14 day recovery period. At the 

### end ofthe 

### recovery period, 

### all animals were eutbanized and necropsied
. Animals were observed twice daily for mortality and moribundity. Clinical signs were recorded daily. Body weight and 

### food consumption were measured weekly. Functional observational 

### battery and motor activity evaluations were perfonned during weeks -1 (pretest), 

### 3, and 

### 5 (recovery). Samples for hematology (42 day) necropsies. Complete necropsies were perfonned on 

### all rats. The brain, liver, kidney, heart, spleen, 

### testes and 

### epididymis or ovaries, adrenal 

### glands, and thymus from 

### all animals were weighed at each necropsy. Approximately 40 tissues were collected and**p~rved** 

### at each necropsy from 

### all animals. 

### The following tissues were examined microscopically from the 

### control and high dose animals: liver, kidney, heart, spleen, 

### testes (males), prostate (males), seminal vesicles (males), epididymis (males), ovaries (females), adrenal glands, thymus, bone with marrow (sternebra), brain, stomach, cecum, duodenum, ileum, jejunum, lymph node, peripheral nerve (sciatic), spinal cord, lung, trachea, uterus (females), urinary

-14-

# •

•

• wn.-186004 CMA BFR.IP bladder, and 

### all gross lesions.) The lungs, liver, kidneys, stomach, gross lesions and target organs were examined in 

### all dose levels. Survival was not affected by administration of the test article. 

### All animals sunrived to the scheduled necropsy. Clinical signs observed during the study were nom..pecific, low in incidence, non-dose-reJated and not considered related to test article. Body weights, weight gain and food**consump~:on** of treated animals were compared statistically by sex and treatment day to their respective control groups 

### (p<0.05 or 

### p<O.on. Body weight, weight gain and food consumption was not affected by treatment. No statistically significantdifferen 'eS in body weightbetween control and treated animals were detected with the exception ofan increase in mean female body weight in the 

### 350 mg/kg/day during week 2 oftreatment. Mean female body weight at that time point was 19ti g vs 179 in**u,~.-** control group. No statistically significant differences in body weight gain between control and treated animalswere detectedwiththeexception ofa decrease in mean male body weightgain in the 1000 mg/kg/day recovery group during week 1 ofrecovery. Mean male body weight gain at that time point was 21 g vs 31 in the control group; mean male body weight was not statistically different from the control mean. No statistically significant differences in food consumption between control and treated animals were detected with the exception of an increase in mean female food consumption in the 

### 350 mg/kg/day during weeks -1, I, and 2 of treatment. Mean female food consumption at that those time points were 18, 17 and 17 g vs 16, 15 and 15 gin the control group, respectively. Functional observation battery and motor activity results from treated animals were compared statisticallyby sex andtreatmentdayto theirrespectivecontrol groups 

### (p<0.05). Theseparameters were not affected bytreatment with the test article. No statistically significantdifferences were observed between treated and control animals at any time p.:;int. No statistically significantdifferences between treated and control animals were found for hematology parameters with the exception of an increase in the mean
-15-

# •

•

• WJL-186004 C'MABFRJP
activared partial 

### tbromboplastin 

### time in tbe 

### 1000 mglkg/day males on week 4 and a decrease in the mean prothrombin time in the 

### 1000 mglkg/day females on week 4. These statistical differences were not of

### toxicological significance. No toxicologically significant effects on serum chemistry values related to test 

### article administration were observed at the 

### 28 day prima. y

**a~d** 

### 42 day recovery necropsies. Scatteredinstances ofstatistically significant differences between treated 

### and controlanimals were detected

### for someserum chemistryparameters at

### the 

### 28 day primary 

### necropsy. These 

### scattered 

### statistical differences were not considered toxicologically significantbecause the 

### statistical differences occurred: in 

### the absence ofa dose response, in the absence ofthe accompanying clinical chemistry changes expected, 

### in the 

### opposite direction from what occurs in a toxic state, 

### in 

### a direction which is without physiologic significance, or due to potential 

### interference with the laboratory method. No statistically significant differences in serum chemistry parameters were detected 

### between groups at 

### the 42 day recovery necropsy. No gross lesions which could be 

### attributed to the test 

### article were detected at 

### eithernecropsy. Gross lesions were nonspecific, low in iucidence, non-dose-related and 

### considered incidental. Nomicroscopic lesion which could be 

### attributed tothe test

### articlewere detected
on histopathologic exam. Microscopic changes were nonspecific, low in incidence, non-dose-related and considered incidental. No statistical significant differences in 

### organ weight or organ to body weight ratios were detected between control and trt:ated 

### animals with one exception. Absoluteliverweights were statistically significantlyincreased with respect 

### to 

### control
means at 

### the 

### 28 day necropsy in 

### males in the 

### high dose and females in 

### the mid and high dose. Liverto 

### body weightratios in mid and 

### high dose males and low, mid and high dose females were statistically significantly increased at the 28 day necropsy. At 

### the 

### recovery necropsy, male absolute and liver to 

### body weight ratio were statisticallycomparabletothe

### control mean whereas female absoluteliverweights and Hver to 

### body weight ratio were statistically significantly increased with respect to control mean. The difference in 

### absolute liver weight between control and treated


### -16-

# •

•

• WIL-186004 

### CMABPRIP
females was lesspronounced

### attheendoftherecovery period, indicatingtheincrease 

### in liver weight was reversible in**f~** as well as males. In the absence oftest articlerelated histopathologic and serum chemistry changes, increases inliver weight areconsidenc.:**~<Il** adaptive, ratherthan a toxic response, arenotuncommon intherat, and are most likely the result ofmicrosomal induction. In conclusion, no systemic toxicity was obseiVed at any dose 1evel. Based on the results ofthis study, the NOAEL (No ObseiVed Adverse Effect Level) ofHBCD administered orally to male and female rats for 28 consecutive days was 1000 mg/kg/day
.

-17-• , r ,( ! )\r ' ' '- I • • - • ..**~** f' 1 •**~** ' ,.**~** l_ r ' •?**.~** • • / -l,.: >I: 

# •

•

• WIL-186004 CMABF'BJP 

## n. OBJECTIY.E Tne objective 

### of the 

### study 

### was to 

### evaluate the 

### possible toxic effects 

### of HexabromocyclododecaDe (HBCD) 

### when administered 

### orally (gavage) to rats 

### for 

### 28 days. 'Ibis

### included evaluation 

### ofpoteDtia1 

### neurotoxicity by function.'ll observational battery (FOB) and 

### motor activity (MA) 

### assessment. Reversibility, persistence 

### or delayedoccurrenceoftoxic effects

### was evaluatedby a 14-daypost-treatmentrecovery period. Tbe 

### selected 

### route ofadministration was 

### oral by 

### gavage as 

### this ro•tte is 

### an acceptable and standard 

### method for 

### administering test 

### article per OECD 

### Guideline 407. Tbe 

### animal model, tbe 

### Sprague-Dawley**Crl:~R** 

### rat, is 

### recognized as 

### appropriate for subchronic 

### toxicity studies and was**se~ected** 

### based 

### on the availability of

### historical control data
.



### -18-

# •

•

• WIL-ltsOllH 

### CMABFRIP


## m. STIJDY DESIGN Acclimation and Pretest Period 

### I Assignment ofAnimals to Study Groups 

### I 12 Males, 12 Females/Group (1 and 4) and 6 Males, 6 Few_>Jes/Group (2 and 3) Treated Daily for 28 Days 

# I Body Weight!' and Food Intake

Recorded Weekly, Clinical Observations Recorded Daily 

# I Functional Observational Battery and Motor Activity Evaluations Petformed at Appropriate Intervals
Clinical Pathology Evaluations on 

### All Animals Prior to Scheduled Necropsy

Necropsy Examinations Performed on

6 Animals/Sex/Group (1-4) At the End ofthe Dosing PeriGd; Selected Organs Weighed; Selected Tissues Mi Necropsy Examinations Petfonned on 

### 6 Animals/Sex/Group 

### (1 and 4) After a 14-Day Recovery Period; Selected Organs Weighed; Selected Tissu Mic

-19-

# •

•

• < ' 

### -**~,** 

## .**~ ~**  .. <" ; - ' 

# . '

WIL-1Bti004 

### CMABFRIP
IV. 

## EXPBRIMBNTAI. PR.OCEPURES A. IND,ODl]CDON 

### 1"beexperimental SlUtdate,

### initiatingwith test

### articleadministration, was May 17, 1996 (week 0). 

### 1"be primary 

### necropsy was on June 14, 1996. The recovery necropsy was on June 28, 1996. 

### 1"be experimental tennination date, concluding with the last

### pathology examination, was 

### October 

### 8, 1996. B. 

## TSST AND CONIROL

### ARDCiffl 1. TEST 

## ARDCLE IDENTIFICA]JQN Hexabromocyclododecane (HBCD) 

### was received from 

### Chemical Manufacturers Association via Wildlife International Limited, Easton, Maryland, as follows: Label Identification #3577HBCD Approximately 300 

### g (Composite"' of#3462, 3519, and 3551) Prep. date: 2/19/96 

### Exp. date: 12/16/96 [Wn. Log No. 3014A] HBCD 

### Test substance no. 3577 Composite* ofno's 3462, 3519, 

### 3551 Prep. date: 2/19/96 Expires: 12/16/96 [WIL

### Log No. 3014B] No. of Containers BFceived One Bottle 

### Gross 

### weight 

### 646.3 g

One Bottle 

### Gross weight 430.1 g 

### Description White powder

White powder Date 

## Received 4/17/96

617/96

"' The test article was a composite sample ofequal 

### portions ofcommercial HBCD manufactured by CMA BFRIP members. The stability and 

### purity ofthe 

### bulk test 

### article were the 

### responsibility of the 

### sponsor. Forpurposes ofdose calculations, the 

### te..q article was considered to be 

### I00% 

### HBCD. The bulk test 

### article was stored in 

### a sealed container in 

### a 

### dry, well-ventilated area 

### at room temperature (68°F 

## ± 4°F) and was 

### -20-

# •

•

• WIL-186004 CMABFRIP
considered 

### to be stable under

### these conditions. 

### C.1 April19, 1996, a 

### 1.013 g sample was taken from tile first shipment of test arncie and stored 

### in the archives at 

## wn. Research laboratories, Llc. An approximate one 

### gram reserve sample ;vas 

### to be taken from tlle second sfa]-iment of test article in 

### accordance with 

## wn. standard operating procedures. However, after the remaining test article had been returned 

### to the**spo~sor,** it was discovered that a retention sample was not collected from this shipment. This deviation 

### from the protocol had no impact on the scientific validity of the study as both shipments were ofthe same lot number. 2. CONTROL ARTICLE IDENI'IFICATION The vehicle control article utilized in preparation ofthe test mixtures and for adtrinistration to the control group was**Mazola~** com oil manufactured by Best 

### Foods, CPC International, Inc., Englewood Cliffs, New Jersey. 

### 3. PREPARATION A sufficient amount of com oil 

### to dose the control 

### grour**~·nimals** was placed in a labeled storage container. The vehicle control article was stirred continuously throughoutthe sampling and dosing procedures using 

### a magnetic stir bar and stirplate. The appropriate amount of test article, HBCD, for each group was weighed into a tared, precalibrated storage container. A**st~** bar was added and the container

### was tared a second time. An appropriate volume ofvehicle was added and the weight ofthe formulation was recorded. 'The formulations were stirred continuouslythroughoutthe sampling anddosingprocedures using magr>etic stir bars and plates. All dosing formulations were prepared daily and stored at room temperature. Test article concentration and homogeneity of ·£he dosing formulations were verified using gravimetric determination. 

### 4. ADMINISTRJ\TION The test mixtures were administered ora1ly by gastric intubation via a 16- gauee stainless steel gavage cannula (Popper and Sons, Inc., HydePark, New


### -21-**~** 

## - ' 

# -**~,** "" - ,... . ... a , , • 

### ., -- ) -**~** . :....."'- .

# . ..

, :.. 1 '
. \ , - '
. .**~io** .- • '**~**

• f**~ ~**  "

.. • - "'

• I -'t> "' • • ' • •**~ ~**  ,. If II • J ' 

# •

•

• WB.-186004 CMABFRIP
York 11040) 

### as a single daily dose for 

### 28**con~ve** 

### days through the 

### day prior

### totbe scheduledprimary necropsy. A dose

### volume of

### S mllkgwas used for all 

### dosage levels. 

### 1be concurrent 

### control group animals 

### teeeived the 

### vehicle, com 

### o:U, on 

### a compuable regimen 

### at a dose 

### volume of 

### S mllkg. 

### Individual dosages were adjusted 

### based on the 

### most recent body weights, to provide tbe comet mglkg/doay dose. 

### On May 28, 1996 (study day 11), all 

### males and the 

### first female in 

### the 3SO mglkg/day group were inadvertently 

### dosed using 

### the dose volumes for all males 

### and the 

### first female in tbe 

### 125 mglkg/day 

### group, and on June 7, 

### 1996 (study day 21), tbe first four males in 

### the 

### 1000 mglkg/day group 

### were inadvertently dosed using tbe dose volumes for the first four males in 

### the 3SO mglkg/day group. All 

### arumab that were underdosed received a conecting dose amount. In the 

### 3SO mglkg/day group, three 

### males (nos. 

### S028S, S0286 and 

### S031S) were overdosed by 0.1 u.l andone female 

### (no. S0322) was overdosed by 0.03 mi. One 

### male (no. S0283) in tbe 1000 mglkg/day group was overdosed by 0.2 mi. 

### 'Ibis deviation from tbe protocol 

### bad 

### no impacton

### the outcome of

### the study as all animals received tbe correct 

### dosing &alution 

### and only a 

### limited number of 

### animal!i received a slightly bigber 

### than intended dosage. 

### The 

### following table presents tbe 

### study 

### group assignment. Dosage Do.qge Number Group Test Level Concentration Volume ofAllimals 

## -Number Artisle <mg/b/dayl (mglml) Cml/lsgl 

## »!Is!**~** 1 2 3 4 

# s. Vehicle 0 0 

## s 12 12 HBCD 125 2S 

## s 6 6 HBCD 350 70 

## s 6 6 HBCD 1000 200 

## s 12 12 

### SAMPLING 

### AJIIP ANALYSES Priorto the

### initiation ofdosing, sample dosing suspensions were prepared for Groups 2 and 4, 

### and duplicate 1 mi aliquots were withdrawn from tbe middle stratum 

### of each suspension immediately after preparation. The suspensions were 

### stilTed overnight using magnetic stir 

### bars 8.i.ld plates, and duplicate 1 

### m1 aliquots were wit!tdrawn from the 

### middle stratum 

### of each


### ·-22-'I • .. • •• \ '

.. j

.. ... • ... • .. - - "r ' <!; • - ),.

- _._. : •**~'·~~'** • .. • "

- •' I .J ' ' # <I; ", • • < - - " o !
.. . ;, . .**-~** ... . .**Ill~** • )

...**~** •**..~~t** ; ........ . a . '"'
.. •., /"
.. 

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### ;..**~** " . 

### . ' ::...

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# •

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### WIL-186004 CMABFRIP su-;pension approximately 2_. hours following p:reparation. The wnples \Vere frozen lU soon as possible after collection and were shipped on dry ice to Albemarlt: CorporationTechnical Cen·er, Baton Rouge, Louisiana, on May 9, 1996, for stability analysis. Duplicate one-gram samples of the test article were collected ;md frozen on the first and last days of**t~~** article administration, and were shipped on dry ice to Albemarle Cot¥->ration Technical Center on May 20, 1996 and Junt; 17, 1996, respectively, for stability analysis. The results of**th~** <JJalySF;S 

### are presented 

### in Appendix A. Gravimetric assays of the dosing formulations for verification of test article concentration and homogeneity v1ere perfonr.ed 

# '"t WIL Research Laboratories, Inc. Concentration Jetenninations 

### v:ere performc..1 daily prior tod"seadministration. Homogeneitydeterminations wereperfunneci on study days 0, 13 and 27. The results ofthese analyses are presented in Appendix B. C. ANIMAL RECEIPr 

### ANI.> ACCLIM<\TIQN/PRETEST PERIOD Forty male and forty female Sprague-Dawley**Crl:~R** rats were received
on May 2, 1996, from Charles River Laboratories, Inc., Portage, Michigan. The animals were 28 days old upon receipt. Each animal \/as examiiJed by a qualified technicianontheday ofreceipt. Animals wereuniquely identified byMonel metal eartags displaying the pennanent identification nua;!:ler. Ail**anim~.s** were housed for

### a 15-day acclimation andpretestperiod; observations 

### v<...re madetwice daily

### for mortality and general changes in appearanc.e or behavi"r. Pretest data collection began on May 

### 6, 1996. bdividual body weights we;e recorded and detailed physical examinations were perfoTJlled on the fu-n and last days of pretest week -1 (May 10 and 16, 1996, respectively). Jndividual food consumption was measured for the interval week -1 to week 0. For computer entry, pretest data were assigned to computer**pro~ol** number WIL-186004P. Pretest clinical observations**~** presented in Appendix C. D. ANIMAL HOUS!'NG Upon arrival, all animals were housed three per cage**~y** sex for a minimum
orthree days. Thereafter, allanimals were housed individually 

### inclean, wire-mesh

-23-

# -

# •

•

• wn..-186004 CMABFRIP
cages suspended above cage-boaJd. All 

### animals were maintained by 

### the animal husbandry staff

### in accordance with the "Guidefor

### the Care and UseofLaboratory Animals" 1• The 

### animal facilities 

### at WlL Research Laboratories, Inc., are accredited 

### by 

### the American Association for Accreditation 

### of Laboratory Animal 

### Care (AAALAC). 

## E. Dmr. DRINKING WATER. AND 

## MAINTENANCE The 

### basal ration used in this study 

### was Purina• Certified Rodent ChowGD #5002. 

### The diet utilized at 

### WlL Research Laboratories, Inc., is a certified feed with appropriate analyses performed by the manufactun·1· and provided to 

## wn. Research Laboratories, Inc. Municipal water supplying thefacility is 

### sampled and analyzedforcontaminantsaccordingtoStandanlOperating

### Procedures. Theresults oftheseanalyses are main;.ained 

### at

### WlLResearch Laboratories, Inc. Contaminants were not present 

### in animal feed or water at levels sufficient to interfere with the objectives of 

### this study. The basal 

### diet and drinking 

### water**d~livered** 

### by 

### an automatic watering system were provided ad

### libitum throughout the study period e:1rc, ·:. •i"ring the period of fasting prior to blood collection when food but not**W'~k** ·. Na.: V**~thheld,** 

### F. 

## ENVIRONMENTAL CQNDI110NS 

### All animals were housed throughout the acclimation period and 

### during the
temperature 

### at 72° 

## ± 4°F (22° 

## ± 2°C), relative humidity at approximately 30- 70% anda minimum of 10 air

### exchangesperhour. Room temperature anarelative humidity were recorded twice daily. Actual 

### recorded ranges for temperature and humidity were 71.9° to 73.0°F (22.2° to 22.8°C) and 

### 33.7% to 67.3%, respectively, during the study period. Light timers were set 

### to provide a 12-hour ligbt/12-hour dark 

### photoperiod. G. ASSIGNMENT OF

## ANIMALS TO

## TREATMENT GROUPS On

### May 16, 1996 (thedaypriortotest

### article adminimation), all available rats were weighed and examined 

### in detail 

### for physical abnormalities. Animals

### judged suitable for testing, 

### as determined 

### by 

### the study director, were selected for use in


### -24-

. . • . •• .**.~** /. -,**(.~j ~-**  

### .-I (._

. . . '

.**-~.** 

### .. '. · . -· . · I 

## l ! I · ... 1- I ! • '

•**~** .. <"• •**~** • ,, ..)**~** - "'·" •

# •

•

• wn.-186004 CMABFRIP
the computerized randomization procedure. The individual body weights were entered into the wn. Toxicology Data Management System (WTDMS"'). A printoutcontainingthe animal numbers, co:rrezponding bodyweights an..i individual group assignments was generated based on computer-generated random number pennutations. The animals then were arranged according to the printout. The control and 1000 mg/kg/day groups each consisted of 12 males and 12 females. The 125 and 350 mg/kg/day groups each consisted of six malec: and six femaies. Afterrandomization into study groups, the animals were then randomized intotwo study replicates to allow for the reasonable conduct of the FOB and locomotor activity assessments. Each dose group and sex were equally represented within each study replicate. Animal no. 50292 was replaced by animal no. 50289 orJ study day -1 as animal no. 50292 died shortly after being handled for pretest clinical obsetvations .?..nd weighing. The selected animals were approximately six weeks old at the initiation ofdosing; body weight values ranged from 159 to 222 grams for the males and from 121 to 161 grams for the females. Several animals weighed less than the protocol-specified minimum weight (175 g for males, 125 g for females) at the initiation of dosing. This deviation bad no impact on the outcome ofthe study as all animals were within the prot'"iCOl-specified age range (4-8 weeks) at the initiation of dosing. H. 

### CI.Thi1CAL OBSERVATIONS AND StJRvTvAL The animals were observed twice daily, once in the m0ming and once in the afternoon, for mortality and moribundity. Clini':al observations were perfonned
on all animal> at the time ofdosing and approximately one to two hours following dosing (designatedas onehourpost-dosingtorreportingpurposes). Duringthe 

### !4- day recovery period, clinical observations were performed daily on the remaining six animals/sex in the control and high dose groups. Each animal received a detailed physical examination prior to the scheduled necropsy. I. BODY WEIGHTS Individual body weights were measured and recorded weekly beginning one week prior to test article administration (week -1). Mean body weight changes

-25-

# •

•

• WIL-186004 CMABFRIP

were calculated 

### foreacb study week. A final body weight was recorded for each 

### animal on 

### tbe day 

### ofscheduled necropsy.**1.~** Individual 

### food consumption was measured weekly beginning one week prior totest

### articleadministration (week

### -1). Foodintakewas calculated

### as g/animallday for the corresponding body weight intervals. K. FUNCUQNAI. OB$ERVADONAL BA'ITER.Y 

### Functional observational battery (FOB) 

### evaluations were performed on six animals/sex/group during the pretest week, the fourth week of test 

### article administration and the last 

### week of the recovery period (weeks 

### -1, 3 and 

### S, respectively). lbesame six animais/sexinthecontrol and 1000mg/kg/day groups were used for each evaluation intetval. Testing was performed by the same technicians without knowledge of the animal group assignment. lbe FOB 

### was performed in a sound-proofed room equipped with 

### a white noise generator set 

### to operateat70 

## ± 10db, withthefollowingexception. Homecageobsetvations were performed in 

### the animal room. All 

### animals were observed for the following parameters 

### as described 

### below (refer to Appendix 

### D for a detailed description of 

### the scoring criteria used for eacb obsetvation): 1. HOME CAGE OBSERV ATIQNS 

### Posture ConwisionstTremors Feces consistency 

### '2. 

### HANDIJNG OBSERVATIONS 

### Ease ofremoval from 

### cage Lacrimation/Cbromodacryorrbea Piloerection Palpebral 

### closure Red/Crusty 

### deposits Eye prominence



### -26- Biting Paipebrai 

### (eyelid) ciosure



### Ease ofhandling animal in band Salivation Fur

### appearance Respiratory rate/character Mucous membranes/Eye/Skin color Muscle tone

# •

•

• wn..-186004**CY'~** BFRJP
3. OPEN 

### f!F1 n OBSERVATIONS (evaluated over a 2 minute observatic.n period) Mobility Rearing Convulsions/Tremors Grooming Biza.rre/Stereotypic behavior Time to first step (seconds) 4. SENSORY OBSERVATIONS Approach response Startle response Pupil response Forelimb extension Air righting refle.>.. 

### 5. NEUROMUSCULAR OBSERVATIONS Hindlimb extensor strength Hindlimb foot splay 6. PHYSIOLOGICAL OBSERVATIONS Catalepsy Body temperature L. LOCOMOTOR ACTIVITY Gait Arousal Urination/Defecation**G~tscore** Backing

Touch response Tail pinch**re~ponse** Eyeblink response Hindlimb extension Olfactory orientation
Grip strength-hind and forelimb Rotarod performance
Body weight

Locomotor activity observations were performed on six animalsisexigroup during the pretest week, the fourth week oftest article administration and the last week of the recovery period, (weeks -1, 3 and 

### 5, rrspecti.'!ely). The same six animals/sex from the control and 1000 mg/kg/day groups were used for each evaluation interval. LocomotorActivity, recorded afterthecompletionofthe FOB, was measured automatically using the Digiscan 'Micro' Animal Activity System (Omnitech Electronics, Inc., Columbus, OH 43228). This is a personal computercontrolled system consisting of individual test chambers which utilize a series of infrared photobeams surrounding a clear plastic, rectangular cage to quantify an animal's motor activity. The testing of treatment groups was done according to replicate sequence. The activity system was operated in the Stagger Start mode of -27-' " -**~** .,.,**t-~ ~....**  ...... \:.. • \**~** ' '**~J** ' • • • < '

) • : • ... T'"->
.. -- .. 

# •

•

• WD..-186004 CMABFRIP operation. 

### Data were collected in one-minute epochs and the test 

### session was 41 minutesindlr.ration foreachanimal. 

### Animal placementintotheactivity cagein the Stagger Start mode of operation 

### initiated the data 

### collection process. The first 

### epoch was oftenincomplete duetotheplacementofthe

### animal in theactivity cage. Forthis 

### reason, the first 

### minute ofdata 

### was deleted. 

### The remaining 

### 40 minutes of data 

### collection 

### (4, 10 minute subsessions) were compiled for tabular presentation. Locomotor 

### Activity was divided into 

### t:wo categories: total and 

### ambulatory activity. Total motor activity was defined as a combination of fine motor skills 

### (i.e. grooming; interruption ofone ortwo adjacent photobeams) and 

### ambulatory motor activity (interruption oftluee 

### or more consecutive**photobeam~).** 

### Refer to Appendix 

### E for additional details on the motor activity methodology. 

### M. CLINICAL PADIQLQGY Blood samplesforclinicalpathologyevaluationsweretaken from thevenacava of all animals just prior to tile scheduled necropsy. The animals were 

### fasted overnightpriortonecropsy and

### thecollection ofblood samples. Clinicalpathology methods, procedures 

### and references are presented in Appendix 

### F. 1. IIBMATOLQGY 

### 1befollowing hematologic evaluations were conducted on blood samples collected from all animals 

### at the scheduled necropsies: Total Leukocyte Count (White 

### Cell) Erythrocyte Count (Red 

### Cells) Hemoglobin Hematocrit Mean Corpuscular Volume (MCV) Mean Corpuscular Hemoglobin

(MCH) Mean Corpuscular Hemoglobin

Concentration (MCHC) Platelet Count Platelet Estimate• 

### RBC Morphology-


### • = Presented only on individual tables 

### ( ) = Designates tabular abbreviation


### -28- .Differential Leukocyte Count -Neutrophil 

### -Lymphocyte -Monocyte -Eosinophil -Bascphil 

### Prothrombin Time (Pro 

### Time) 

### Activated Partial Thromboplastin



### 1ime (AP'IT)

# •

•

• wn.-186004 CMABFRIP
2. SERIJM CHEMISTRY The 

### following serum chemistry evaluations were conducted on blood samples collected from all 

### animals at the 

### scheduled necropsies. Glucose Blood Urea 

### Nitrogen Blood Creatinine Total 

### Protein Albumin Albumin/Globulin Ratio

(A/G 

### Ratio) Globulin Calcium Phosphorus Chloride Total 

### Bilirubin (Total Bill) 

## ( ) = Designates tabular abbreviation N. 

### PATIIOLQGY 1. 

### MACROSCOPIC EXAMINATIQN Gamma Glutamyltransferase (Glutamyl Transfer) Serum Aspartate 

### Aminouansferase

(Aspa.rtat 

### Transfer) Serum Alanine AminotransferaSP.

(Alanine 

### Transfer) Serum Alkaline Phosphatase

(Alkaline Phos'tse) · Sodium Potassium Total Cholesteroi 

### (Cholesterol}

A 

### complete necrorsy was conducted on all 

### animals that 

### were euthanized atthe end ofthe dosin6 period 

### orfollowing a two week recovery period. All 

### unimals were euthanized by cartx:m dioxide asphyxiation and exsangumated. The necropsy included, but was not limited to, 

### examination of the external 

### surface, all 

### orifices and the cranial, 

### thoracic, abdominal and pelvic cavities 

### including 

### viscera. At the time of necropsy, the following tissues and 

### orga_n_s were collected and preserved in 10% neutral buffered formalin:

-29-

# •

•

• WIL-186004 CMABFRIP 

### AdreDals {2)* 

### Aona 

### Bone witb marrow {sternebrae)* 

### Bone marrow 

### smear (from 

### femur)• Brain (fonbrain, midbrain,

hindbrain)* 

### Byes witb optic nerve (2) 

### GastrointestiDa tract



### Esophagus 

### Stomach* Duodenum* 

### Jejunum* Ileum* 

### Cecum• 

### Colon Rectum Heart* 

### Kidneys (2)* Liver (sections oftwo lobes)* Lungs [including bronchi, fixed by

inflation with fixative (2)]* Lymph node 

### (mediastiul)* Lymph node 

### (mesenteric)* Mammary gland 

### (females only) 

### Ovaries with oviducts 

### (2)* Pancreas Peripherzl 

### nerve (sciatic)* PituitaJy Prostate* Salivary glands [submaxillary (2)] 

### Seminal 

### vesicles (2)* Skeletal muscle (vas'rus medialis) Skin Spinal cord 

### (cervical, midthoracic,

lumbar)* Spleen* Testes with epididymides (2)*" 1bym":Js* Thyroids [with parathyroids if



### present (2)] 

### Trachea* Urinary bladder* 

### Uterus with vagina* All gross lesions•
• = 

### Bone marrow 

### smears were obtained atthe

### scheduled necropsy butwere not

### placed in**10~** neutral 

### buffered formalin. " = Tbe testes 

### and 

### epididymides were preserved in Bouin's solution. 2. ORGAN

## WElGHTS 1be

### following organs from animals euthanized at

### the week 4

### primary and 

### week 

### 6 

### recowry necropsies were weighed: Adrenals 

### Brain Epididymides Heart 

### Kidneys Liver Ovaries Spleen Testes Thymus Paired organs were weighed together. Organ 

### to final 

### body weight ratios were calculated. 3. MICROSCOPIC EXAMiiiADON 

### After 

### fiXation, specified tissues (designated 

### with an asterisk 

### in Section IV.N.l.) were sent

### to Colorado Histo-Prep, 

### Inc., 

### Fort Collins, Colorado, for
-30-

# •

•

• wn.-186004 CMABFRIP trimming and staining (hematoxylin-eosin). All tissues from the control and high dose group animals designated with an asterisk in Section IV.N.1 andthe lungs, liver, kidneys, stomach and gross lesions from the low and mid dose g:oup animals were examined microscopically. Microscopic examination of these tissues was conducted by Carney B. Jackson, D.V.M., B.S. An. Sci., D.A.C.V.P., D.A.C.V.P.M., Assistant Director ofPathology and Veterinary**Medicin~.** 

## wn.. Research Laboratories, Inc. 0. STATISTICAL ME'lliODS All analyses were conducted using two-tailed tests for minimum significance levels of I % and 5% comparing the treatment groups tc the vehicle control group by sex. All means werepresented witb standard deviations (S.D.)andthe numbers of sampling units (n) used to calculate the means. All statistical tests were performed by a Digital* MicroV**~** 3-'!rJ() computer with 

### appropri.ate programming. Analysis ofbody weight, body weight cbange, food consumption, clinical pathology values, continuous functional observational battery (FOB) data and absolute and relative organ weight data were subjected to a one-way analysis of variance, followed by Dunnett's tesf. Discontinuous (ordinal or descriptive) FOB data were analyzeJ using Fisher's exact 

### tesf. Statistical tests for locomotor activity data were performed using a personal computer installed with SAS/STAT statistical software3• Clinical labo·'itory values for cell types that OCCI!r l'!t a !ow incidence (i.e., monocytes, eosinophils and basophils) were not subjected to statistical analysis. P. DATA RETENTION The sponsor will b..:-. title to all documentation records, raw data, specimens or other wor'· product ce:;er..ted during the performance ofthe study. All work product induding raw pt.per data and specimens will be retained in the archives at Wll.. Research laborn;ories, Inc., as perprotocol. Raw <bta in m:>gnetic form, a retention sample of the test article and the originaJ final 

### report will be retained 

### in the archives atWll.. Research Laboratories, Inc., in compliance with regulatory requirements
.
-31- 

# -

# •

•

• WIL-186004 CMABFRIP 

### V. Bffltn.TS A. CUNJCALOBSJmVADONS 

## AND SURVIVAL 

### Summary 

### Data: Tables 1, 2, 3, 4, 

### 5 Individual 

### Data: Tables 57, 58, 59, 60, 61 AU animals survived tothe 

### scheduled**~ies.** 

### Noclinic:al signs wbich couldbe attributed tc

### the test 

### a.-ticle were observed at the 

### clinical examiutions performed 

### at the 

### time of dosing, one hour following dosing, 

### during the 

### recovery period 

### orpriorto the 

### scheduled necropsies. Clinical 

### findings in the treated 

### groups occulTed similarly 

### in the 

### control group and/or in a 

### limited number ofanimals. No relationship to the test 

### article was evident. 

### B. BQDY

## WEIGHTS Summary 

### Data: Tables 6, 7 

### Individual 

### Data: Tables 62, 63 Mean body weights in the treated 

### groups were statistically comparable to the 

### conttol 

### group means throughout the 

### dosing and 

### recovery periods, 

### with the 

### following exception. Mean body weight in the 

### 350 mglkg/day

### group females was significandy (p<O.OS)increased

### onweek2. Meanbody weightgains inall treated 

### groups were statistically comparable to the 

### control group values throughout the dosingperiod. 

### Duringthe

### recoveryperiod

### (weeks

### 4-5 and5-6), mean body weight gain 

### in tile 

### 1000 mg/kglday group males was reduced 

### (statistically significant at 

### p<0.01 for week 4-5) in comparison to the 

### male control group mean. However, these

### decreases were 

### not attributed

### to

### treatment as no

### te.~ article-related effects on body weight were noted 

### during the 

### dosing period. 

### Inthe 

### 1000 mglkg/day group females, mean bodyweightgains

### were statistically<'.omparabletothe 

### control

### group values during the 

### recovery period. 

### C. FOOD CONSUMPUQN 

### Summary Data: Table 8 Individual 

### Data: Table 64 Food consumption, evaluated as g/animallday, was not affected 

### by test 

### article administtation 

### at any dose leve!. 

### The only statistically significant (p<O.OS or**-3~-**

# •

•

• WD..-186004 CMABFRIP 

### p<O.Ol) differences from the control group were slight increases in food consumption 

### in the 

### 125 and/or 

### 350 mglkg/day group females during the 

### frrst two weeks oftestarticle administration. However, thevalues**~·ere** comparableto their respective

### pretest values and decreases, :rather than increases, in food consumption are generally considered to be toxicologically significant; no relationship to treatment was evident. All other food consumption values in tht> treated groups were similar to the control group values. D. FUNCTIONAL OBSERVATIONAL BATTERY 

### 1. HOME CAGE OBSERVATIONS Summary 

### Data: Tables 

### 9, 10, 11, 12, 13, 14 Individual 

### Data: Tables 65, 

### 66, 67 Historical Control Data: Appendix G No remarkable differences were apparent between the control and treated groups when the home cage observati..Jns were evaluated during study weeks 

### -1 (pretest period), 

### 3 and 

### 5 (recrvery period)
. 2. HANDliNG OBSERVATIONS Sum'llary 

### Data: Tables 

### 15, 16, 17, 18, 19, 20 Individual Data: Tables 

### 68, 69, 70 Historical Control Data: Appen<Hx G No remar'ICable differences were apparent between the control and treated groups when the handling observations were evaluated during study weeks ··1 (pretest period), 

### 3 and 

### 5 (recovery petiod). 3. QPEN FIELD OBSERVATIONS 

### Summary Data: Tables 

### 21, 22, 23, 24, 25, 26 Jndividual Data: Tables 

### 71, 72, 73 Historical Control Data: Appendix G 

### N.:. remarkable differences were apparent between the control and treated groups when the open field observations were evaluated during study weeks -1 (pretest period), 

### 3 and 

### 5 (recovery period)
.


### -3j-

# •

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• WD..-186004 

### CMABFRIP
4. SENSORY OISBIYA'DONS 

### Summary 

### Data: Tables 27, 28, 29, 30, 31, 32 Individual 

### Data: Tables 74, 75, 76 Historical Control 

### Data: Appendix 

### G No remarkable differences wme 

### apparent between 

### the control and treated groups 

### when the

### sensorimotorobservations**we~e** 

### evaluatedduring study weeks -1 (pretest 

### period), 

### 3 and 

### S (recovery period). 

### S. NBlJROMUSClJLAR OBSERYA'UONS 

### Summary Data: Tables 33, 34, 35, 36, 37, 38 Individual Data: Tables 77, 78, 79 Historical Control Data: Appendix 

### G No test**article-~eJated** 

### diffeieDces were apparent between the control and treated

### groups when the

### neuromuscular

### responses wereevaluated during study weeks -1 (pretest 

### period), 3 and S 

### (recovery period). 1be 

### only statistically significant (p<

### 0.01) differences from the control group v.ere decreased 

### hindlimb grip strengths 

### in the 

### 125 and 

### 1000 mglkg/day group females at the 

### week 

### 3 evaluafon. However, similar values were observed for these 

### groups duringtheprer t 

### evaluation andthe 

### 1000mglkg/daygroup

### female mean at

### the week S 

### recovery evaluation was comparable to 

### the control group value; therefA'e, 

### the decreases 

### could not be 

### attributed to the test 

### article. 6. 

### PHYSIOLOGICAL OBSERVADQNS 

### :Summary Data: Tables 39, 40, 41 Individual Data:**Tabl~** 

### 80, 81, 82 Histor;caJ Control Data: Appendix 

### G No remarkable differences were apparent between the 

### control andtreated 

### groups when the 

### physiologicalparameters were evaluated during study weeks -1 (pretest 

### period), 

### 3 and 

### S (recovery period)
.



### -34-

# •

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• WR.-186004 CMABFRIP


### E. LOCOMOtOR 

## ACDVlTY Summary 

### Data: Tables 42, 43, 44 Individual Data: Tables 83, 84, 85 Histoncal Control Data: Appendix H Noremarkable differences in mean ambulatory and total motoractivity values in the 

### 125, 350 and 

### 1000 mg/kg/day 

### group males and females were observed during study weeks 

### -1 (pretest period), 

### 3 and 

### 5 (recovery period). 

### F. CLUnCALPATHOLOGY 

### 1. IIEMATOLQGY Summary Data: Tables 45, 46, 47 Individual Data: Tables 

### 86, 87, 88, 89, 90, 91 No test article-related effects on hematology parameters were observed at the week4primaryorweek 6 recovery evaluations. A slight increase in mean activatuf partial thromboplastin time (APTI) in the 1000 mg/kg/day group males was statistically significant 

### (p<0.05) when compared to the control group value at the week 4 evaluation. However, APTT increases of this magnitude are not considered to be toxicologically significant. A statistically significant 

### (p<0.01) decrease in meanprothrombin time occurred in the 

### 1000 mg/kgiday group femaies at the week 4 eval•Jation. However, increases,

### ratb~r than decreases, in prothrombin time 

### are gener ..!ly considered to be toxicologically significant. No other remarkable differences in hematology parameters were observed. 2. SERUM CHEMISTRY Summary Data: Table 48 Individual Data: Table 92, 93 Historical Control Data: Appendix I Noadverseeffects relatedtotestarticleadmini!.trati0n on ser._.•.**chemi~** parameters were observed at any dose level at the 28 dayprim2.ry necropsy or the 42 day recovery necropsy.

-35-**1Un_1G~ ~~~** **~~~-~"""~** 

### CMABFRIP 

# •

•

• Scattered instances 

### of statistically significant differences between treated andcontrolanimals were

### detected for some serum chemisuypuameters

### atthe 28 day 

### primary oecropsy. These scattered statistical differences were not ooosideredanadverse

### effectof

### thetestarticlebecausethestatisticaldifferences 

### occumd: in 

### the 

### absence of a dose 

### response; in the absence of the accompanying clinical chemistry changes expected (i.e. t!te increase in total pJasma protein in mid and high dose males and high dose females was not accompanied by an 

### increase in hematocrit and total red blood cell count numbers); in 

### the opposite direction from what occurs in a toxic state (i.e. the increase in globulin and albumin/globulin ratio in mid dose males and 

### the increase inalbumin in high dosefemales wherea decrease in 

### these parameters 

### is indicative of toxicity); in a direction which 

### is without physiologic significance (i.e. the decrease BUN and creatinine in mid dose females, the decrease in serum 

### alanine aminotransferase and :.lSp&Itate aminotransferase 

### in low, mid andhigh dosemales andmid

### and high dosemales, respectively); due to**interf~** with

### the laboratory method (i.e. theincrease in ser ..m chloride in mid and high dose males and in low, mid and high dose females

- free bromine in 

### the test Miele can artificially elevate chloride by reacting in the laboratory method as freechlorideanions). Additionally, althoughdetected as statisticallydifferentfrom concurrentcontrol values, theabsolutechangeinthe measured parameters were small and within 

### the range of·historical 

### ct'lgtrol valuesfor

### this species, strain, sex and laboratory (i.e. total**plasm~**protein

### and the albumin/globulinratio 

### inmiddosemales, cholesterol in mida."ld high .iose females, serum glucose in high dose females, serum calcium in high dose females). No statistically significantdifferencesin serumchemistryparameters were 

### detected between groups at 

### the 42 day recovery necropsy.



### -36-

# •

•

• CMABFRIP
G. 

## PAlHOLOGY 1. :MACRQSCOPIC EXAMINATiON Silmmary 

### Da1a: Tables 49, 

### 50 Individual Data: Table-: 

### 94, 95 Nc test article-reJated gross lesions were**obser-.~ed** 

### at the week

### 4 primary necropsy oratthe week 6 recovery necropsy. Gross lesions inthe controi and treated groups occurred with similar frequency or were observed in single animals. 

### 2. ORGAN WEIGHTS Summary Data: Tables 

### 51, 52, 53, 54 Individual Data: Tables 

### 96, 97, 98, 99 No statistical significant differences in organ weigh!: or organ to body weight ratios were detected between control and treated animals with one exception. Absoluteliverweights were statistically significantlyincreased with respect t{'l control**mearo~** at the 28 day primary necropsy in 

### males in the high dose a..-td females 

### inthe mid and high dose. Liverto tody weight ratios in mid and high dose males and low, mid and high dose females were statistically significantly

### mcreased atthe 28 day necropsy. At

### the recovery necropsy, 

### maY: absolute and liver to bcly "';eight ratio were statistically comparable to the control 

### mean at the recov<!ry**n~rop~y** whereas female absolute liver weights and liver tc oody weight ratio were statistically significantly increased with 

### respo.,"t to control mean. The dll:erence in absolute liver weights between control and treated females wa.s less 

### pronuunced at 

### the end of the recovery period, indicatingtheincrease in livex**wei~bt** 

### was reversib!e in females 

### as well 

### as males. In the absence of test article related ilistopatho!ogic and serum chemistry**chang~,** increases inliverweight are considered an adaptive, rather than a toxic response, are not uncommon in the rat, and are most likely the result of

### microsoma.t induction
.

-37-

# •

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• WIL-186004 CMABFT.dP


### 3. MICROSCOPIC EXAMINATION 

### Summary Data: Tables SS, S6 Individual 

### Data: Tables 94, 

### 9S No test

### article-related histopathological lesions were observed at 1\e week 4primary orweek 6recovery necropsies. Lesions noted 

### in 

### the treated 

### groups occUlTed 

### in a limited 

### number of animals, 

### in a 

### manner wbicb was 

### not doserelated, or

### with similar frequency in 

### the 

### control group.

-38-

+**~1..** ' •**~** •**~** - ....**~** - '

- l-1

- ' "-
.... ....... t ,, ' ' '. ' 

### I I

. I j . /. 

### I I' 

### I '

. 1-**~** t ,._ ' b**~** - \ • •**~** • --:::-**~** ..:...-"**~** -

# •

•

• WlL-186004 CMABFRIP
VI. DISCUSSION

### AND CONCLUSIONS All animals survivedto scheduled necropsy. Noclinical signs related 

### totestarticle administration were observed. Notestarticle-related effectsonbodyweight, bodyweightgain, food consumption, functional observation

### battery orlocomotorartivity werefoundineithersex atany dose during either the 28 day dosing interval or during the 14 dz.y recovery period. NotestarticJe-related adverseeffectsonserum chemistryorhematologyparameters were detected during the study. No test article-related gross or microscopic lesions were found at any dose level in either sex at either necropsy. Absolute and relative organ weights were unaffected by test article administration with the exception ofli·1er weight. Absolute liver weight was statistically increased at the 28 day necropsy in

### rnales ofthe high dose and in females at the mid and high dose; relative liver weights were increased ii1 the mid and high dose males and in low, mid and high dose females. At the day 42 recovery necropsy, male absolute and relative liver weight were statistically comparable to the control mean. Female absolute and relative liver weight was statistically increased relative to the control mean 

### at the recovery necropsy. This difference was less pronounced than at the 28 day necropsy, which indicates theincrease in female liverwas reversible as was the male liverweight. Increases in liver weight in the absence of test article-related changes in serum chemistry and hlstopathoiogy 

### are likely the resuit ofmicrosomal induction, which is an adaptive response in tlte rat and not evidence oftoxicity. In conclusion, no evidence of systemic toxicity was observed at any dose level or time point. Based on the results of this study the NOAEL (No Observed Adverse Effect Level) ofHBCD administered orally to male and female rats for 28 consecutive days was 1000 mg/kg/day.
engelis, Ph.D., D.A.B.T. tudy Director

-39- 

### 13 

# F;6 '!) Date

# - 

# •

•**~** ---........**~** - ,...-**,.~~** !
. •**~** .,.# ' ' ' J - ''- '**-~-** • ' t ' ' • ' .,.. /!"' > "-- "' "-.. I - .-**-~** 

### WD.-186004 CMABFRIP

VB. 

## ICBY PP$0NNBJ,

## AND REPQRT SUBMISSION Study 

### Stmmisog: Kerin Clevidence, B.S. Sally A. 

### Keets, A.S. 

### Gary R. Kiplinger, 

### B.S. Ken D. 

### Miller, B.A., M.T.(ASCP) Daniel W. Sved, 

### Ph.D.

ieyiewed By: 

### Group Supervisor ofGross Pathology and Developmental Toxicology Laboratory 

### Maaager ofVivarium Mauager 

### ofGeneral Toxicology Senior Groups Supervisor

- Clinical Pathology 

### Director ofMetabolism and Analytical 

### Chemistry



### Carney B. , D.V.M., B.S. An. 

### Sci., D.A C.V.P., D.A.C.V.P.M
. 

### Assistant 

### Director ofPathology and Veterinary 

### Medicine Stu logist



# ' .s. lmecbllr'ofToxicology

lis, 

### Ph.D., D.A.B.T. 

### ·orToxicologist Study 

### Director 

# d-l:.laz

_ Date


### I) 

# &6 1') Date

# •

•

• wn..-186004 CMABFRIP


### vm. PAJHOLQGISI SUMMARY STATEMENT Study animals werepre-designated for eutbanization either 

### atthe end ofthe dosing period (week 4) or following a two-week recovery period (week 6). All animals were euthanized by carbon dioxide asphyxiation and exsanguinated. A complete macroscopic examination was perfonned on each study animal that included, but was not limited to, the external surfaces, all orifices and cranial, thoracic, abdominal and pelvic cavities including viscera. No test article-related external or internal findings were noted 

### at the week 4 frlmary orthe week 6 recovery necropsies. SpP..:cified tissues (refer to section IV.N.1.) were preserved in 10% neutral buffered formalin and prepared of microscopic evaluation by Colorado Histo-Prep, Inc., Fort Collins, Colorado. Microscopic examination ofthe tissues was performed by Carney B. Jackson, D.V.M., B.S. An. Sci., D.A.C.V.P., D.A.C.V.P.M., Assistant Director of Pathology and Veterinary Medicine, 

## wn.. Research Laboratories, Inc. Findings (lesions) in the test aWcle-treated groups occurred either in a limited numberofanimals, 

### in a manner which was not dose-related ar with similarfrequency in the control group. In conclusion, under the conditions ofthis study, no test article-related macroscopic or microscopic fmdings (lesions) were apparent at any dose leve! at the week 4 primary and week 6 recovery necropsies.

# 13~!7 Carney B. ackson, D.V.M., B.S. An. Sci., Date D
..C.V.P., D.A.C.V.P.M. Assistant Director of Pathology and Veterinary Medicine Study Pathologist

-41- 

# -

# - .. . "'"; '

- 

# . . . '**~** I -**~** • ' '**~** ! • I**~ ~**  • I 

# •

• WD..-186004 CMABPRIP 

## . , "' '



## IX. OUAI.lTY ASSJJIANCE

## UNIT SIATBMENI

Date(s) of Pbue 

### Ipsnestjgp<(l 

### Ip=Jed 5/17/96 Teat MaterW Prepanlioomel Allalylis 5/W96 ADimiJ 

### c.e " Equipment 6/11/96 FUDCtioaal Oblervatioaal Battery 6/14196 Blood Collectioo & Allalylis 6/14/96 Necropsy 9124196 Study Records Vol. 1-1 9123, 24, 26, 27196 Study Records Vola. 1-2 & 1-3 9124196 Study Records Vol. 1-4 9125196 StudyRecords Vol. Rx-1 9125196 Study Records Vol. N-1 9126196 Study Records Vols. C-1, 2, 3 10/10/96 Study Records Vol. HIP-I 10/14/96 Draft Report (Appendix B ODly) 9/26, 10/3, 4, 6, 7,

14, 20, 21196 lmft Report (w/oAppeDdix B) Date(s) F'mdiop Reported to Study

## Directer

5/17/96 5128196 6/11/96 6/14/96 5/14/96 9126196 9127196 9126196 9126196 9126196 9126196 10/10/96 10/14/96 10/21/96 Date(s) FiDdiDp Reported to M•p•geppt
6120196 6120196 1130196 1130196 7130/96 10/24/96 10/24/96 10/24/96 10/24/96 10/24/96 10/24/96 11/26/96 11/26/% 11126196


### 1bestudy wu**condo~** 

### and inspectedinaccordance with theUnited StatesEPAGood Laboratory Pm.'tice Regulations, the OBCD Principles of 

### Good Laboratory Practice, the Standatd Operating Procedures of

# wn.. Aesearcb Laboratories, Inc., and the protocol. Quality Assurance fmdings, derived from 

### the inspections 

### during the conduct !lf the study 

### reponed to 

### the study director. A status report is submitted to management monthly. 

### 1beraw data, retention sample(s), 

### ifapplicable, and the final report 

### in the Archives 

### at 

## wn, Research Laboratories, Inc.' oranother location specified by the sponsor.

# •

•

• wn.-186004 CMABFRIP



# Colorado Histo-Prep, Inc. Post Oflicc Box 270733 • Fon Collins. CO 80527 • 970-493-2660 CH-P Study # 1948 Client's Study #WIL-186004 QUALITY ASSURANCE S·IATEMtNT This study has been conducted in accordance with the**t~A** Health Effects Test Guidelines (40 CFR Part 792) and the OtCD GLP's, C(81) 30 (Final) Annex 2, including appropriate SOP's and designated recording**re~~irements.** The study •as inspected by a member of the Quality Assurance Unit and reported to the Project Director and Management as recorded below.**Comp~risons** were made between the raw data and the final report, and they were found to be in agreement. All records were reviewed to assure**tha~** any problems found during the course of a Quality Assurance Inspection were corrected and properly documented. PHASE REVIEWED Tissue receipt in file Protocol reviewed Tissue list compared to protocol Animal numbers,**~Px,** necropsy dates reviewed Trimming Embedding Microtomy Slides compared to blocks**SliQ~5 CO!!!par~d**  to blocks Blocks compared to inventory Wet tissue compared REVIEWED BY Aimee Kramer Aimee Kramer Aimee Kramer Aimee Kramer Aimee Kramer Aimee Kramer Aimee Kramer Aimee Kramer Aimee Kramer Aimee Kramer to inventory Aimee Kramer Handwritten forms checked for writeovers Computer forms compared to CH-P animal forms Necropsy sheets compared to animal forms Slides returned Slides returned Blocks returned Wet tissue finalized for shippment Ai:tr""e Kramer Aimee Kramer Aimee Aimee Aimee Ai.mee Kramer Kramer Kramer Kramer Aimee Kramer



### -43- OAT£ REVI.EWED 07-12-96 07-11-96 07-11-96 07-12-96 07-15-96 07-19-96 07-25-96 07-25-96 07-30-96 07-30-96 10··18-96 07-29-96 07-30-96 07-2S.-96 07-26-96 07-30-96 10-13-96 10-18-96 PROJECT DIRECIOR 10-18-96 10-18-96 10-18-96 10-18-96 07-15-96 07-19-96 0'/-25-96 10-18-96 10-18-96 10-18-96 10-18-96 10-18-96 10-18-96 10-18-96 MANAGEMENT 10-18-96 10-18-96 10-18-96 10-18-96 07-15-96 07-19-96 07-25-96 10-18-96 10-18-96 l0-18-96 10-18-96 10-18-96 10-18-96 10-18-96

# •

•

• WIL-186004 CMABPIUP



# Colorado Histo-Prep, Inc. Pcllll Office Boa 270733 • Fort Collins. CO IO:o
. 1 • 97o-493·2660
The raw histology data consistinq of paper records related to this study are archived at Colorado Histo-Prep, Inc., located at 319 Lincoln court, Fort Collins, 

### co 80524. Raw data of blocks, slides, and wet tissues are temporarily stored at Colorado Histo-Prep, Inc. until forwarded to client. All raw Quality Assurance data are archived at Colorado Histo-Prep, Inc., Fort Collins, co 80524. Storage is in accordance with the EPA and OECD quidelines•



### -44-

\--
- - I t \ • 0 •

# •

•

• -- , , f • • ' r - •**~-** I - . •**~** ' I - • /

WIL-186004 CMA BFI{D)
X. 

### REFERENCES 1. National Research Council (1996) Guide for the Care and Use of Laboratory 

### Animals, Institute ofLaboratory Animal 

### Resourr---..s, Commission on 

### Life Sciences. National Academy Press, Washington, D.C. 

### 2. BMDP (1979) Biomedical Computer Programs. (Dixon, W.J. and Brown, M.B., eds.) University ofCalifornia Press, Berkeley, CA, 

### pp. 612, 780, 781. 3. SAS (1991) SAS/STATUser's Guide, Varsion 6, 4th Edition. SAS Institute, Cary, North Carolina, 1028 pages.



### -45-