COD:ING FORMS FOR SRC INDEXING

Microfiche No.

OTS0572612

86940001508

06/23/94 07/22/94 8D

CHEM MFGS ASSN

EASTMAN KODAK CO

ETHYLENE GLYCOL MONOBUTYL ETHER: ACUTE ORAL TOXICITY STUDY IN

THE GUINEA PIG WITH COVER LETTER DATED 07/21/94

Chemical category

ETHYLENE GLYCOL MONOBUTYL ETHER (CASt 111-76-2)
REV. 7/27/82

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Document Processing Center (TS-790)

Room L-100

Office of Pollution Prevention and Toxics

U.S. Environmental Protection Agency

401 MStreet, SW

Washington, DC 20460

RE: TSCA Section 8(d) Health and Safety Report Rule

(Notification/Reporting); 40 CFR 716.60

Dear Sir or Madam:

~~9110<:x::)' s o~

1b

8"9lfooolSo~ 0iEMICAL. MANUFACTUAERS ASSOCIAllON

July 21, 1994

.,.,

-.. .' ---.

- ,

..!

Pursuant to TSCA Section 8(d) Notification/Reporting requirements, The

Chemical Manufacturers Association (CMA) submits the following

information on initiation and/or completion of research on ethylene

glycol monobutyl ether, (EGBE), CAS Number 111-76-2. CMA is submitting

this information on behalf of the U.S. manufacturers of EGBE

who are the research sponsors. The manufacturers/sponsors are:

Eastman Chemical Company

P.O. Box 431

Kingsport, TN 37662

Shell Oil Company

One Shell Plaza

P.O. Box 4320

Houston, TX 77210

Union Carbide Chemicals and

Plastics Company

39 Old Ridgebury Road

Danbury, CT 06817

Dow Chemical Company

2020 Willard H. Dow Center

Midland, MI 48674

Occidental Chemical Corp. S Greenway Plaza

Suite 2400

Houston. TX 77227-7702

The following research is reported on:

1. Development of a Physiologically-Based Pharmacokinetic Model for

Hu.an Health Risk Assessment .... .

'~~urpose of this research is to acquire data that will Allow

the construction of a reliable physiologically-based

pharmacokinetic .odel for EGBE so that appropriate risk ass.s~nt

prooedures can be carried out. The research is being conducted at

the Toxicology Research Laboratory, The Dow Chemical Complny,

Midland, MI. This research is being conducted over a multi-y.ar

period and began in 1991. An 8(d) notice of initiation was

submitted to EPA on March 20, 1991. A report of the initial phase

of the research is submitted with this letter. This project

continues.

250' M Street. NW. •....asI i1lQlD'1. DC 20037 TfJI8PI'1One 202-887-' '00 Fax 202-887-1237 Lc-.:= R' c.. "i' c; •
Docu-ent Processing Center

July 21, 1994

Page 2

2. Studies of the Effects of EGBE and Butoxyacetic Acid on Normal

and Abnormal Human Erythrocytes

The purpose of this research is to examine the effects of EGBE and

its principal metabolite, BAA, on normal and abnormal human and

rodent erythrocytes to further elucidate a possible mode of action

for EGBE-induced hemolysis in the rodent. The research is being conducted at the Department of Medicine, Section of Hematology, Baylor College of Medicine, Houston, TX. This research is being

conducted over a multi-year period and began in 1990. An SCd)

notice of initiation was submitted to EPA on January 17, 1990.

Two publications arising from the first phase of research are

submitted with this letter. Work on this project continues.

3. Dermal Bioavailability Study in Fasted Volunteers

The purpose of this research is to assess the dermal

bioavailability of EGBE vapor by an arm-only exposure. The

research is being conducted at L.A.B. Bio-Research International

Inc., Senneville, Canada. The research began in April 1994 and

should be completed in the fourth quarter 1994.

The Panel has also recently conducted three studies that are exempt from SCd) reporting. As these studies may be of interest to EPA, we

are submitting information on them as follows:

4. Acute Dermal and Oral Toxicity in the Guinea Pig

The purpose of these studies was to determine the oral and dermal

toxicity of EGBE in male and female guinea pigs. The studies were

carried out at the TOXicological Sciences Laboratory, Eastman

Kodak Company. The studies began in March 1994 and were completed

in June 1994. Final reports for the acute dermal and acute oral

toxicity, respectively, are submitted with this letter.

5. Acute Vapor Inhalation Toxicity· StudY in Guinea Pigs

The purpose of the study was to determine the acute inhalation

toxicity in guinea pigs resulting from a single I-hr exposure to

EGBE vapor. The study was carried out at Bushy Run Research

tenter, Export, PA. The study began in March 1994 and was

co-pleted in July 1994. The final report is su~itted with this

.fetter.

-, -- If yoa-have any questions regarding this submission, please contact me

at 2021887-1293.

Regards,

~-:!fM~ Manager

Ethylene Glycol Ethers Panel

Enclosures
-

86948881588

STIJDY lIfLE

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ETHYLENE GLYCOL MONOBUTYL ErBER

ACUTE ORAL TOXICITY STUDY IN THE GUINEA PIG

HAEL NUMBER: 94-0300 KAN: 902270

CAS REGISTRY NUMBER: 111-76-1

FINAL REPORT

AUTHOR

Kenneth P. Shepard, B.S.

PERfORMING LABORATORY

Toxicological Sciences Laboratory

Corporate Health and Environment Laboratories

Eastman Kodak Company

1100 Ridgeway Avenue

Rochester. New York 14652-6272

USA

LABORATORY PROJECT ID

9403OOAO

STIJDY SPONSOR

Ethylene Glycol Ether Panel

Chemical Manufacturers Association

2S01 M Street, NW

Washington, DC 20037

CMA REFERENCE NUMBER

EGE-S8.o-GPIG-EASTMAN

STUDY COMPLIDON DATE

June 23, 1994
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QUALITY ASSLTRANCE INSPECIJQN STATEMENT

. [21 CFR S8.3S(B)(7). 40 CPR 792.35(B)(7), and 40 CFR 160.35(B)(7)]

STUDY: 94-0300-1 STUDY DIRECTOR: SHEPARD,K.P.

ACCESSION NOHBER: 902270

PAGE 1

04/20/94

STUDY TYPE:

};./.

(AUDITOR, SURANCE UNIT)

'lO THE BEST OF MY KNOWLEDGE, THIS FINAL REPORT ACCURATELY DESCRIBES

'rIlE HE'l'BODS AND S"l'ANDARD OPERATING PROCEDURES, AND THE REPORTED

RESULTS ACCURATELY REFLEcr THE RAW DATA. THIS STUDY WAS INSPECTED

BY 1 OR MORE PERSONS OF THE QUALITY ASSURANCE UNIT OF HAEL, EASTMAN

KODAK COMPANY ROCHESTER, N. Y. AND WRITTEN STATUS REPORTS WERE

SUBMITTED ON THE FOLLOWING DATES:

------------------------------------------------------------------

INSPEC'l'ION

DATES

PHASE(S)

INSPECTED

STATUS REPORT

DATES

03/01/94 PROTOCOL SUBMISSION

TEST SYs-rEM RANDOMIZATION

TEST SYs-rEM IDENTIFICATION

TEST SYSTEMS WEIGHTS

DOSE CALCULATIONS

TEST ARTICLE MEASURE AND MIX WITH CARRIER

TEST ARTICLE-cARRIER-MIXTtJRE DOSING OF TEST

SYS'l'EMS

04/13/94 GROSS PATHOLOGY

PATHOLOGY REPORT

04/20/94 FINAl. REPORT REVIEW

DRAFT REPORT

04/13/94

04/20/94
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COMPLIANCE WITH GOOD LABORATORY PRACI1CE STANDARDS

The study described by this report was conducted in compliance with the following Good

LaboIatoIy Practice Standards:

United Stares Environmental Protection Agency, 40 CFR Pan 792 (TSCA) as revised

August 17, 1989.

United Stares Food and Drug Administration, 21 CFR Part 58 as revised September

4, 1987.

Annex 2 of the Organization for Economic Cooperation and Development Guidelines

for Testing of Chemicals C(81)30 (Final).

~hH Kenneth P. Shepard,••~~~ B.S.

Study Director

drtJ,<' Hll.f M nth/DaylYear

MontblDaylYear
TABLE OF CONTENTS

ABSTRACT

PERFORMING LABORATORY

SPONSOR

STUDYDAl'ES

STUDY DIREcrOR and OIHER. KEY PERSONNEL

PURPOSEIOBJEC'TIVE

MATERIALS AND MEI'HODS

Test Article O.arac:terizatiOD and Analyses

Test Procedure Guideline

Test System

Isolation

Bousmg and Environmental Conditions

Diet and Water

Animal Ideotification

Randomization

Dose Level and Dosing Regimen

Control Substance and Vehicle

Oinical Observations and Body Weight Determinations

Necropsy

RESULTS

DATA ANALYSIS

DISCUSSION AND INTERPRETATION

CONCLUSION

DATA STORAGE

SIGNATURE PAGE

APPENDIX TABLE OF CONTENTS

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Page Number'

S

6

6

6

6

7

7

8

8

8

8

9

9

9

.9

9

10

10

10

17

17

17

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ABSTRACI

ETHYLENE GLYCOL MONOBUTYL El'HER

ACUTE ORAL TOXICITY STUDY IN THE GUINEA PIG

BAa NUMBER: 94-0300 KAN: 902270

CAS REGISTRY NUMBER: 111-7~2

An oral toxicity study was initially conducted in five male and five female guinea pigs

administered a single limit dose of 2000 mg/kg of the test article by gavage. Following 60%

mortality for males and 100% mortality for females at the limit dose, a single oral dose of

either 1000 or 500 mg/kg of the test article was administered to groups of five male and five

female guinea pigs. Mortality was 20% for both sexes at 1000 mglkg and 0% for both sexes

at SOO mg/kg.

Abnonnal clinical signs included slight to severe weakness and prostration that were either

transient or noted prior to death. Other abnonnal clinical signs indicate that the test article

was probably irritating to .the stomach. These signs included sialorrhea and staining of the

hair of the face and/or abdomen by feed and saliva and were initially noted approximately 48

hours after dosing and for only the two surviving males at the 2000 mg/kg dose level. In

addition, gasping and a red nasal discharge were noted for a single male at the 1000 mg/kg

dose levd on the day following dosing. These two abnormal signs persisted to Day 3 of the

study wbeo. this male died. No other abnormal clinical signs were evident during the 14-day

observation period. All guinea pigs which survived to termination of the study g.ined weighL

The primary treatment-related cbanges noted at necropsy confirmed that the test article was a

gastrointestinal irritant. These included necrosis and hemorrhage in the gastric mucosa and

the accumulation of mucoid contents in the duodenum. No tissues were collected for

histological examination. No evidence ofred blood cell toxicity or hemolysis was observed

during the study. The ID5C) for guinea pigs was calculated to be 1414 mglkg with a 95%

confideDce level of 1020 to 1961 rng/kg for the combined sexes.
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PERFORMING LABORATORY

Toxicological Sciences Laboratory

Corporate Health and Environment Laboratories

Eastman Kodak Company

1100 Ridgeway Avenue

Bo320 Kodak Park

Rochester, New York: 14652-6272

USA

SPONSOR

Ethylene Glycol Ether Panel

Chemical Manufacturers Association

2501 M Street, NW

Washington, DC 20037

Authorized Representative: Kathryn A. Rosica

STUDY DATES

Study Initiation: March 1, 1994

Experiment Initiadon: March 1, 1994

Experiment Completion: March 29, 1994

Study Completion: JW1e 23, 1994

STUDy DIRECTOR.

Kenneth P. Shepard, B.S.

OTHER KEY PERSONNEL

Len Sakal, B.S•• Study Technician

John W. Mosher, B.S., Principal Investigator

Milan S. VIaovic, D.V.M., Ph.D., Laboratory Animal Medicine, Pathologist
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The purpose of the study was to determine the oral toxicity of the test article in male and

female guinea pigs.

MATERIALS AND METHODS

Test Article Characterization

Test Article NamC".:

Synonym:

CAS RegistIy Number:

CMA Reference Number:

Ethylene glycol monobutyl ether

2-Butoxyethanol

111-7~2

EGE-58.o-GPIG-FASTMAN

HAEL Number:

Laboratory Project Number:

Kodak Accession Number:

Soun:e of the Test Article:

94-0300

940300AO

902270

Eastman Chemical Company

Kingsport, TN 37622

Physical State and AppeaI3Dce: Colorless liquid

Lot Number: Lot 9792

Received at Performing Laboratory: February 17, 1994

Test Article Analyses

The test article was analyzed for purity using gas chromatography with flame ionization

detection (GCIFID). Samples of the test article was collected prior to initiation of the study

and after termiDation ofthe in-life phase of the study. Both the initial and final samples for

purity were determined to be 99.8% ethylene glycol monobutyl ether. Based on these dam.

the test article was considered to be stable throughout the study. The initial test article sample

was also analyzed for struetu:Ie confirmation using gas chromatography with mass

spectlometry (GCIMS). The structure of the test article was consistent with the structure of

ethylene glycol monobutyl ether based on comparison with reference library spectra.

Concentration verification of the dosing solutions consisted of analyzing each of the three

concentrations f~", 20%. and 10%) of the test article in distilled water. The solutions were

analyzed on the day of dosing, or as soon as possible thereafter, using gas chromatography

with flame ionization detection (GCIFID) under the same instrument conditions as used for
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MATERIALS AND MEIHODS. COQL

Test Article Analyses. Cont.

purity determination. The overall mean measured concenttations for the 40", 20". and 10"

solutions were 42.5%. 18.2". and 10.1" (weight/volume), respectively. All analyses were

perlormed by the Chemical Quality Services Division. Kodak Park. The analytical test report

is presented in the appendix.

Test Procedure GuicfeJinelReference

OECD Guideline for Testing of Chemicals: Guideline 401 (Annex V, Test B.1); also in

accordance with the Department of Transportation. CPR Title 49, Section 173.132, except

guinea pigs were selected for the test species.

Test System

Species/Strain:

Source:

Arrived at Performing Laboratory:

Number of Animals:

Sex:

Body Weight Range

at Study Initiation (grams):

Age at Study Initiation:

Isolation

Hanley Guinea PiglCrl:(HA)BR VAFIPIlJS'8

Charles River Laboratories, Kingston, NY, USA

February 16, 1994 (for 1000 and 2000 mglkg groups)

March 9, 1994 (for 500 mglkg group)

Five of each sex per dose group

Male and Female

Males = 309 - 405 Females = 310 - 375

Males = 5-7 weeks Females = 5-7 weeks

Animals were isolated and monitored for at least five days after arrival and before release to

the testing facility.

All animals were individually housed in suspended. stainless-steel, mesh cages.

EnyjronrnenJal Conditions

A photoperiod of 12 hoUl'S light from 6 a.m. to 6 p.m. W2S maintained. Room tempe:ratuIe

was maintained at 69-72 'F. Relative humidity was maintained at 57-59".
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Mt\1lJUA1$ AND METHODS. Cont, .

J2je:t iDd Water

Agwa~ Prolab- Animal Diet (RMH 3000) certified pellets and water (Monroe Co\Dlty (NY)

Water .t\utbority) were available ad. libitum. No known contaminants which would interfere

with the outeOllle of this study were expected to be present in feed or water from these

soura:s. Analyses of feed and results of quarterly analyses of water are maintained on file

within the testing laboratory.

AP.inmI Identification

All aniJnals were identified by cage numbers and uniquely-numbered, metal ear tags.

Cage NUiIlbers:

Cage Numbc:rs:

Cage NUI11bers:

Males

161-165

176-180

201-205

Females

166-170 (2000 mglkg dose group)

186-190 (1000 mglkg dose group)

206-210 (500 mg/kg dose group)

A clinical examination was performed on each animal to ensure that only healthy animals were

utilized. The procedure for including animals in this study was to randomly select and assign

animals from the same shipment to each dose level. Randomization was done by computergenera.ted lists using the Automated Animal Toxicology System. After assignment of animals

to the study, the body weights were determined to ensure that individual body weights did not

exceed 20% ofthe mean weight for each sex.

Doses of 2000, 1000, or 500 mg test articlelkg body weight were administered to the animals•

.t\ single dose of the teSt article was administered by gavage to animals that had been fasted

overnight.

C9ntro1 S,ybstance

No control substance was used.
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MATERIAlS AND METHODS. Cont,

Vehicle

The test article was dissolved in distilled water. Variability in the volume of test article and

vehicle administered was minimized by adjusting the concentration to ensure a constant

volume at all dose levels.

Clinical Observations

Animals were observed at least three times on the day of dosing and at least once each day

thereafter for the duration of the experiment (a total of 14 calendar days). Observations

included, but were not limited to, changes in the skin; fur, feces; urine; eyes; mucous

membranes; respiratory, ciIculatory, autonomic, and central nervous systems; somatomotor

activity; and behavior pattern.

Body Weight Petenninations

Body weights were collected on Days 0 (prior to treatment), 7, and 14.

Animals that died during the study were necropsied as soon as possible. Surviving animals

were necropsied at the completion of the 14-day observation period.

RFSPLTS

Mortality

Mortality was 60% for males and 100% for females at the limit dose (2000 mglkg). For the

lower doses, mortality was 20% for both sexes at 1000 mglkg and 0% for both sexes at 500

mglkg.
RESTTLTS. Cont,

MORTALITY TABLE

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2000

1000

500

5,5

5,5

5,5

3,5

1,1

0,0

Day 0 to Day 1

Day 1 and Day 3

Dose Volumes TABLES OF DOSE VOLUMES

(Dose Group - 2000 mg/kg)

161

162

163

164

165

388

309

335

387

386

1.94

1.55

1.68

1.94

1.93

166

167

168

169

170

358

357

333

365

358

1.79

1.79

1.67

1.78

1.79
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RFSlJLTS. Coot.

Dose Volumes. Cont, TABLFS OF DOSE VOLUMES. Cont.

(Dose Group - 1000 mglkg)

'~."'="'!.•.'RECEIVED .. ·.·.:.

v

.:O.:(:.·.:·.·~T.·.· ..··. ", N"QMBER :. :~):: UUwJ

176 397 1.99 186 375 1.88

rn

178

179

180

405

399

359

392

2.03

2.00

1.80

1.96

187

188

189

190

350

331

343

368

1.75

1.66

1.72

1.84

(Dose Group - 500 mg/kg)

201

202

203

204

205

334

326

326

326

322

1.67

1.63

1.63

1.63

1.61

206

2m

208

209

210

310

322

319

336

320

1.55

1.61

too

1.68

1.60
RFlillLTS. Coot.

Clinjcal Observations

TABLE OF CLINICAL OBSERVATIONS

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2000 Severe Weakness and Prostration Day 0 SIS Males

(Transient or Prior to Death) SIS Females

2000 Death Day 0 to Day 1 3/S Males

SIS Females

2000 Slight to Moderate Weakness Day 1 to Day 2 212 Males

2000 Sialorrhea Day 2 212 Males

2000 Staining (Feed and Saliva) of the Hair Day 2 to Day S 212 Males of the Face and/or Abdomen

2000 Appeared Clinically Normal Day 6 to Day 14 212 Males

1000 Moderate to Severe Weakness Day 0 SIS Males

(Transient or Prior to Death) SIS Females

1000 Prostration Day 0 4/S Males

(Transient or Prior to Death) 3/5 Females

1000 Death Day 1 115 Females

1000 Appeared Clinically Normal Day 0 (p.m.) 4/5 Males

to Day 14 4/4 Females

1000 Moderate-Severe Weakness,Gasping,and Day 1 to Day 3 115 Males Red Nasal Discharge (Prior to Death)

1000 Death Day 3 115 Males
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RWJLTS. Cont.

Clinical Observations. Cont.

TABLE OF CLINICAL OBSERVATIONS. Cont.

SOO

SOO

Slight Weakness

(Transient)

Appeared Clinically Normal

Day 0

Day 0 (p.m.)

to Day 14

SIS Males

515 Females

SIS Males

SIS Females

At the 2000 mg/kg dose level, abnormal clinical signs included slight to severe weakness and

prostration that were either tIansient (severe weakness and prostration were noted only on the

day of dosing, moderate weakness was noted only on Day 1, and slight weakness was noted

only on Day 2) or noted prior to death. Three of the five males and all females died either on

the day of dosing or the day following dosing. Abnormal clinical signs for the two remaining

males included sialorrhea and staining of the hair of the face and/or abdomen with feed and

saliva. These signs, were noted approximately 48 hours after dosing. Slight to moderate

weakness was also noted for the two surviving males at this dose level. By Day 6 of the

study, the remaining two males appeared clinically normal.

For the 1000 mg/kg dose level, moderate to severe weakness and prostration were noted only

on the day of administration of the test material and were either transient or noted prior to

death. One offive females died on the day following dosing. In the afternoon on the day of

dosing, four offive males and all remaining females appeared clinically normal. Abnormal

clinical signs for the remaining male included modeIate to severe weakness, gasping, and a

red nasal discharge. This male died on Day 3 of the study.

Abnormal clinical signs noted for the SOO mglkg dose group were limited to slight weakness

for all animals on the day of dosing. By the afternoon examinations on the day of dosing, all

animals at this dose level appeared clinically normal.
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RESULTS. Coot.

All anjmals that survived to termination of the study gained weighL

TABLE OF INPMDUAL BODY WEIGHTS - MALES

2000

2000

2000

2000

2000

1000

1000

1000

1000

1000

500

500

500

500

500

161 388 441

162 309 Died Day 1

163 335 Died Day 0

164 387 Died Day 1

165 386 430

176 397 Died Dav 3

177 40S 486

178 399 493

179 359 427

180 392 476

201 334 428

202 326 417

203 326 386

204 326 394

205 322 397

498

512

548

555

469

512

498

486

436

466

430
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RESULTS. Cont.

Boc!y Wei&hts. Cont,

TABLE OF INDIVIDUAL BODY WEIGHTS - FEMALES

2000 166 358 Died Dav 1

2000 167 357 Died Day 0

2000 168 333 Died Day 1

2000 169 365 Died Day 0

2000 170 358 Died Day 0

1000 186 375 469 50'

1000 187 350 416 457

1000 188 331

1000 189 343

1000 190 368

500 206 310

500 207 322

500 208 319

500 209 336

500 210 320

Died Day 1

406 443

427 456

366 405

375 420

406 443

424 454

391 447
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RESULTS. Cont,

NecromY and Histgpathology Emdinp

The primary treatment-related chaDges noted at necropsy included minor to moderate diffuse

necrosis and mjnjmal to minor hemorrhage in the gastric mucosa; and minimal to minor

accumulation of mucoid contents in the duodenum. No tissues were collected for histological

examjnation. A record of the individual animal necropsy findings is presented in computergenerated tables which are included in the appended pathology report.

DATA ANALYSIS

The LDso in male and female guinea pigs:

1414 mglkg (95% C.I. = 1020 to 1961 mg/kg)

No dose/mortality curve was constructed since graphs become statistically useful only with the

use of large numbers of animals and dose groups.

DISCUSSION AND INTERPRETATION

The test. article was a gastrointestinal irritant as evidenced by abnormal clinical signs of

sialoabea and staining of the face and abdomen with feed and saliva and treatment-related

chaDges of necrosis and hemorrhage in the gastric mucosa and the accumulation of mucoid

contents in the duodenum that were noted at necropsy. All other abnormal clinical signs were

either transient (noted only on the day of dosing) or noted prior to death. No evidence of red

blood cell toxicity or hemolysis was observed during the study.

CONCLUSION

The IDso for the combined sexes of guinea pigs was 1414 mglkg with a 95% confidence level

of 1020 to 1961 mg/kg.

DATA STORAGE

All test results presented in this report are supported by raw· data which are maintained in the

archives of the Corporate Health and Environment Laboratories. Eastman Kodak: Company.
SIGNATURE PAGE

Kenneth P. Shepard, B.S.

Smdy Director

.M.D., Ph.D.

th and Environment Laboratories

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'-:::\'--& \c;C1'f__ MontlllDaylYear

L ZZ,IC,~'i

MontlllDay/Year
TABLE OF CONTENTS FOR THE APPENDIX

ANALYTICAL TEST REPORT

PATHOLOGY REPORT

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PAGE

NUMBER

1

19
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APPENDIX

Page 1 of 32

ANALYTICAL TEST REPORT

PUlUTY, STRUCTURE CONP:IlUmT:I:ON AND CONCENTRATION

DE'1'E1UIDm.TJ:ON

OF

Ethyl.ene Gl.ycol. !lODobutyl. Ether

KAN: 902270 HAEL No. 94-0300

Nancy K. Porter

TESTING FACILITY

Environmental Analytical Services

Chemicals Quality Services Division

Eastman Kodak Company

1100 Ridgeway Avenue

B-320 Kodak Park

Rochester, New York 14652-6276

SPONSOR

Toxicological. Sciences Section

Corporate Health and Environment Laboratories

Eastman Kodak Company

1100 Ridgeway Avenue

B-320 Kodak Park

Rochester, New York 14652-6272

COMPLETION DATE: 4q494 AMENDMENT DATE: v.jifJ,l
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APPENDIX

Page 2 of 32

ANALYTXCl\I, QUr.I.rn ASSOIUUICE I:NSPECT'ION S'rJ'.'1'EHEN'1

(CFR 58.35(B) (7) 792.35(B)(7) 160.35(B)(7»

~: 94-o300-Z STUDY DI:RECroR: STAUDENKJ'.YER.Jt..

~OL DDIECTOJl:

IOUI: 902270

CQS JOB 1ItJHBER:

S'l'UDy TYPE: PUIUTY. S'rROcr.OONFDUi. AND CONC. DE'rERIUNATI:ON

~~CE·tJNrr)

7BE E'OLLOWI:NG PBJ>.SES OF 'J:HI:S sraDY WERE I:RSPECrED BY ORE OR MORE PERSONS OF

7BE QOALI:'rY ASStJIUUfCE UHrr ON 'DIE DA:rES LISrED BELOW. WRI:TrEN STATOS REPORTS

'lEIlE SOBKl:'f'TED 1:0 "mE STUDY DDlEcrOR AND APPROPRXA'ft: M»tAGEHENT.

I:NSPEcr REQUEST PImSE(S) STA'rOS REPORr

DA7ES NtIKBER DlSPEClED DATES

03/14/94 PROTOCOL SOBKI:SSI:ON 03/14/94

PROTOCOL~

04/06/94 179342 TEST REPORT I:NSPECTl:ON 04/06/94

179347

179343

179344

179346

PtJRI:Tr. STRUCTURE CONFI:RHATI:OR AND CONC. DETERHI:NATI:ON

04/07/94 179342

179347

179343

179344

179346

TEST REPORT I:NSPEl::'l"l:ON 04/07/94

04/18/94 179342

179343

179344

179346

179347

REPORT AMENDMENT

Page 2 of J.8

04/18/6!J4

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APPENDIX

Page 3 of 32

To the best 01: the sigDer's JciIow1edge and be1ief. the study

described by this report vas conducted in COIIIPli.ance 'lith the

rollowinq Good Laboratory Practice standards:

United states Food aDd DrUg Administration, 21 CFR Part

58 as revised september 4. 1987.

united. states EmriromDental. Protection Agency, 40 CFR

Part 792 ('.rSCA) as revised AugUst 17, 1989_

Annex 2 of the OX'ganization for ECOnomic COOperation and

Development Guidelines for Testing of Cbeai.cal.s

(C(81) 30(FinaJ.) ) as required by council Directive

87/18fF:E.C of December, 1986.

Kathy staudezmayer

study Director

Page 3 of 18

<t/I~f1'i T'

Date

JO>,N: 902270
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APPENDIX

Page 4 of 32

TABLE OF CONTENTS

."

ABSTRACT • • _ _ _ • • _

SPONSOR"S STUDY • • • • • • _ • _

SPONSOR REPRESENTATIVE

STUDY INITIATION DATE.

ANALYTICAL START DATE

TEST SUBSTANCE • _ • _

STUDY DIRECTOR • _ • •

ANALYTICAL PERSONNEL

STORAGE CONDITIONS

HATERJ:ALS AND METHODS •

PORIT'l DETERM:t:NAT:t:ON

STRUCTURE CONFIRMATION _ _ _ _ _

CONCENTRAXION DETERMINATION _ CALCULATIONS • _

EXAMPLE CALCULATION. __

REStlLTS

PURITY • _ • • • - • •

STRUCTURE CONF:t:RMATION _ _ _ •

CONCENTRATION DETERMINATION • DISCUSSION • • • • _ • _ • _ •

DATA STORAGE AND RECORD RETENTION

SIGNATURE PAGE • _ • • • • _ • • •

Page

5

6

6

6

6

6

6

6

6

6

6

7

7

8

9

9 _ .10

• 10

• 12

• 12

• • _ 13

FIGURES:

1 EXAMPLE CHROMATOGRAM FROM PURITY DETERMINATION • • 14

2 EXAMPLE CHROMATOGRAM FROM CONCENTRATION

DETERMINATION • • • • • • • • • • • • _ • • 15

3 EXAMPLE TOTAL ION CHROMATOGRAM FROM STRUCTURE

CONFIRMATION • _ • _ • • _ _ _ _ • • • _ _ • _ • • 16

4 EXAMPLE MASS SPECTRUM FROM STRUCTURE CONFIRMATION 17

REPORT AMENDMENTS • • _ _ • _ • _ _ _ • • _ • _ • • • • 18

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Page 5 of 32

The purity phase of the analYtical support for this study

consisted of an initial and final purity assay of the test

substance. The purity was determined to be 99.8% at the

beginning and 99.8% at the end of the study. A mass spectra1

analysis was also done on the initial sample to confirm the

structure of the test substance.

The concentration verification phase consisted of analyzing

each batch of the test substance mixtures at concentration

levels of 40%, 20% and 10% test substance in distilled water.

The overall. mean measured concentrations for the 40%, 20%, and

10% test mixtures were 42.5%, 18.2% and 10.1% (weight/vol.ume)

respectively. The mean measured concentration for the reanalysis of the 40% dose was 44.5% (weight/volume).

Page 5 of 18 KAN: 902270
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APPENDIX

Page 6of32

SPONSOR. REPRESENTATXVE

Kenneth Shepard

STUDY ~:IAT:ION DATE: 2/23/94

ANALYTZCAL S~ DATE: 2/2S/94

. Name:

HAEL No.:

KAN:

sample Ref. No.:

S'rODy D:IREC'rOR

Ethylene glycol monobutyl ether

94-0300

902270

Lot 9792

Kathy Staudenmayer, Chemist

l\NALYTI:CAL PERSONNEL

Nancy K. Porter, B.S.

STORAGE COND~:IONS

The samples were stored at room temperature in a locked

cabinet.

~IUALS lWD HETBODS

The following procedures were used to obtain the data

presented in this report.

PURITY DETERMINATION

A test solution was prepared by dissolving approxilllately

2.0 g of the test substance into a 100 mL volumetric flask and

diluting with methanol. The test solution was analyzed using

gas chromatography with flame ionization detection (GC/FID)

and the following method:

Instrument:

Column:

carrier:

OVen:

:Injection:

Znj. Temp.:

Detector Temp.:

Hewlett Packard 5890

J+W DBWAX. 30m X 0.2Smm i.d., 0.2Sum film

thickness

Belium at 10psi initial bead pressure

80·C (no hold time) to 17S·C (no final

hold time) at S·C/min

1 ul split with split flow at 60cc/min

27S·C

27S·C

Page 6 of 18 KAN: 902270
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APPENDIX

Page 7 of 32

STRUCTURE CONFIRMATION

A test solution was prepared by dissolving approximately

0.2 g of the test substance ·i.'nto a 10 mL volumetric flask and

diluting with methanol. The test solution was analyzed using

gas chromatography with mass spectrometry (GC/MS) and the

following method:

:Instrument:

COlumn:

carrier:

Oven:

Injection:

Inj. Temp.:

Interface Temp.:

Source Temp.:

Hode:

Hewlett Packard S970 quadrupole MS

J+W DBWAX, 30m x O.25mm Ld., G.1Sum film

thickness

Helium at 10psi initial head pressure

80°C (no hold time) to 175°C (no final

hold time) at s·oc/min

1 ul splitless injection

27S0C

27SoC

250°C

electron ionization (EI)

CONCENTRATION DETERMINATION

Three samples of the test substance in distilled water were submitted for analysis on three separate days. The test

substance mixtures were analyzed using gas chromatography with

flame ionization detection (GC/FID) using the same instrument

conditions as the purity phase of this study. A dose

preparation of 40% (weight/volume) of the test substance in

distilled water was received and analyzed on 3/1/94. An

"al.iquot of the 40% dose preparation was diluted with distilled

water and analyzed. A five point calibration curve (ranging

from 4000 mg/L to 20,000 mg/L) was prepared using the pure

test substance diluted in distilled water, and was analyzed

along with the dose sample.

A dose preparation of 20% (weight/volume) of the test

substance in distilled. water was received and analyzed on

3/3/94. The results were highly variable from injection to

injection of the same dilution; therefore, the data were

considered unreliable (:these data are not reported) and the

method of analysis was modified to inclUde the addition of 1-

m.ethyl-2-propanol as an internal standard. The 20% preparation

was reanalyzed on 3/4/94. The 40% preparation was also

reanalyZed to determine the stability of the test sUbstance in

distilled water. However, due to a technical error, the 20%

and 40% preparations were reanalyzed on 3/5/94 when it was

discovered that the standards were not analyZed on 3/4/94.

All the values from the 3/4/94 and 3/5/94 analyses are

reported.

Page 7 of 18 KAN: 902270
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Page 8 of 32

A third dose preparation of 10% (weight/vol.ume) ·of the

test substance in ctistiJ.l.ed water was received and ana1yzed on

3/1.5/94. The intez:nal. standard (1-methoxy-2-propanol.) was

used with this anal.ysis al.so~ The following dilutions were used: .

SAMPLE :IO ALJ:QUar (mt..) ALIQUOT (mL) FINAL

DOSE :INTERNAL VOLUME

STANDARD (mL)

40% Dose 1.0 NA 25

20% Dose 0.6 0.15 10

40% Dose Repeat 0.3 0.15 10

10% Dose 1.0 0.15 10

CALCULATIONS

* 1CL1 * 100%

1000(mL) * 1(gl

1000 (mg)

Linear regression (used for initial. .(0% samp1e):

y=lIIX+b

y = area m = slope

x = conc. (mg/L)

b = y-intercept

OF = volume sol (mL1

Al.iquot taken (mL)

, sample = conc. from linear reg. curve (mg/L) * OF

lUtE" (used for remaining samp1es)

Definitions:

:IS = internal standard

RRF = rel.ative response factor determined from a ca1ibration

standard

RF = average of the RRF factors determined for the individual.

calibration levels

DF = dilution factor

Page 8 of 18 KAN: 902270
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Page 9 of 32

RRF = area std. * amount IS (mqlL)

amount Std. (mq/L) * area IS

OF = volume sol (mL) * 1. (gl '* 1. (Ll '* 100

A1iquot taken(mL) 1000(mg) 1000(mL)

Amount spl. (wt/vol %) = area spI. '* amount IS '* OF

area :IS '* RF

EXNWLE CALCULATION

RRF for injection U (3/15/94) 4144 (1IlCJfL) std.

RRF = 234389 '* 1.5000(mg/Ll = 1.3687

4144(mq/L) '* 619853

Average RRF for 3 injections of 4144 (1II9'/L) standard = 1.4086

RF = Average RRF for 5 calibration standards = 1.39696

OF for 10% dose sample:

OF = 10(mLl * l(g) * 1CLl '* 100

1 (mL) 1000 (mg) 1000 (mL)

0.001 (gl CLl

(mg) (mL)

10% spl. inj U (wt/vol %) = 605074 '* 15000('mg/Ll '* O.OO1Cgl (Ll

631381'* 1.39696 (mq)(mL)

= 10.3 (g/mL) = 10.3%

RESULTS

PURITY

sample Request for Date Mean std. N:

IO Analysis No. Analyzed Dev.

Initial 179342 2/25/94 99.8% 0.015 3

Final 179347 3/25/94 99.8% 0.012 3

Page 9 of 18 KAN: 902270
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Page 10 of 32

STRUCTURE CONFIRMATION

The structure of the test substance was consistent with the

structure of ethylene glycol monobutyl ether based on comparison witli reference libr.u:y spectra.

CONCENTRATION DETERHrNATION

40% '.rest Hi%ture

Date Ana1yzed: 3/1/94

Request for Ana1ysis No.: 179343

Concentration of the Test SUbstance in Distilled Water

(weight/volume%)

40% Test Mixture

mean = n

(Injection '2 not used)

Injection tJ.

Injection '3

42.5%

2

42.6%

42.4%

20% '.rest Hixture

Dates Ana1yzed: 3/4/94, 3/5/94

Request for Analysis No.: 179344

concentration of the '.rest Substance in Distilled Water

(weight/volume%)

20% Test Mixture

'4 - '6 injected 3/5/94)

18.8%

19.7%

20.9%

17.0%

16.7%

16.0%

Injection '1

Injection '2

Injection '3

Injection '4

Injection '5

Injection '6

18.2%

6

1.92

10.5%

mean

n

Sx

CV

('1 - '3 injected 3/4/94;

Page 10 of 18 KAN: 902270
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APPENDIX

Page 11 of 32

40% ~est Hixture Repeat

Dates Anal.yzed: 3/4/94, 3/5/94

Request for Analysis No.: 1.79344

Concentration of the Test SUbstance in Distil1ed Water

(weight/volume%)

40' Test Mixture

mean

n

SX

CV

=

=

:Injection 'I

:Injection 12

:Injection 13

:Injection '4

:Injection '5

Xnjection 16

44.5'

6

2.12

4.76%

46.6%

47.2%

44.7%

41.8%

44.1%

42.7%

(tl. - '3 injected 3/4/94: '4 - '6 injected 3/5/94)

10% Test Hixture

Date Analyzed.: 3/15/94

Request for Analysis No.: 179346

Concentration of the Test Substance in Distilled Water

(weight/volUJ1le%)

10% Test Mixture

mean

n

SX

CV

=

=

=

=

J:njection '1

:Injection 12

:Injection 13

10.1.%

3

0.51

5.05%

Page 1.1 of 18

10.3%

9.53%

10.5%

RAN: 902270
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APPENDIX

Page 12 of 32

DI:SClJSSI:ON

The purity of the test substance was measured using' gas

chromatography with flame ionization detection. 'Xhe initial.

purity of the test substance was 99.8~ and the final. purity was 99.8:t at the end of the study.

The structure confi.xmation of the test substance was perfo:r:med

using gas chromatography with mass spectrometry. The structure

of the test substance was consistent with the structure of

ethyl.ene gl.ycol. monobutyl. ether based on comparison with

reference l.ibrary spectra. An exampl.e spectrum is shown in

figure 4, along with a reference l.ibrary spectrum of 2-butoxy

ethanol (ethyl.ene glycol. monobutyl. ether).

The concentration verification of the test substance mixtures

in distil.led water were determined foll.owing dilution with

distil.l.ed water. The mean concentrations obtained for the

10,20, and 40~ preparations were 10.1, 18.2 and 42.5,

respectivel.y. Al.though an oversight in the 3/4 analysis of the 20~ preparation was discovered in which the standards were not

injected, the data from the 3/4/94 analyses could not be

discounted because the 40~ preparation which was reanal.yzed at

the same time was within 10% of the val.ue from the 3/1/94

anal.ysis.

DAn STORAGE AND RECORD RETENT:IOH

Al.l original. raw data wUl. be transferred to the Health and

Environment Laboratories archives of the Eastman Kodak Co.,

Building 320, for storage. .

Page 12 of 18 KAN: 902270
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APPENDIX

Page 13 of 32

signature Page

ADLYST._:"'-~J2~···~~,;g.......k~~~u..~.=-.· 4~-'~2_-<:>a_4~_ N<lraCirj. Porter D~

Page J.3 of 1.8 KAN: 902270
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APPENDIX

Page 14 of 32

FIGURE 1

EDHPLB CHR01m'.rOGRlU( PROH PURrrY D~I:ON

SAHPLB OF SUBSTANCE J:N HB'rB]WOL

llut.,cIwoe.

.

1000

900

eao

700

;::

-

.-

400

300

200

100

8.0

. . 4.0 6.0 1.2.0. . 14.0 16.0

.

-

-

eao

700

-

sao

.

-

300

200

100

8.0 2.0 4.0 6.0 "0 10.0 '11_ ...--....,

Page 14 of 1S

U.Q 14.0 111.0

!<AN: 902270
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APPENDIX

Page 15 of 32

FIGURE 2

EDHPLB cmto~~OK COBCENntAttON DE'r~:tON

40% ':rES'r SUBSDNCB HLUuQ Dl:L1J'.rJ:ON wrm D:tSTXLLED

. tm:rElt.

(DlCLVDES S'DNDMm)

"", ..Ietrwo-

...0 "l.O ....0 1••0 l ..O

S20

280

240

:zoo

:- 160 "i 120

110'"

~.O

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APPENDIX

Page 16 of 32

FIGURE 3

BDHPLE ".rO'.rAL :tON cmt.~PROH s-ntUCTQRB CONE':IR!m.TJ:ON

IDHPLB Ol!' 'nIB SUBSTANCE :tN ~OL

Fil. >E8ftU

Page 16 of 18

is .'

DN: 902270
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APPENDIX

Page 17 of 32

fIGURE 4

EXAHPLE HAaS SPEc.rR1JH I'ROH S~UC'rlJRE CONP':IRHA'rXON

Fi1. )E&n£2 904-0300 SPL. 1U1.. 9pl< Ab 21159.SPLLT. 30n Cl9UAX. 0.15Ut1 FILn Scan U-7

SUB 5.86 _in.

57

, 100

'110

REFERENCE X.rspARY IlABS SPEC'rRUH OP'

ETHu.ENE GL~COL HONOBUTYL ftHER

Fil. )91609

BpI< Ab 99')9_

Scan 19751

0.00 .in.

19

,

57

,

100 104 ,./

Page 1.7 of 1.8 DN: 902270
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APPENDIX

Page 18 of 32

Report Amendments

Title: Purity. Structure Confirmation and Concentration

Determination of Ethylene Glycol Monobutyl Ether

KAN: 902270

The following changes were made to the report:

Location: Page 7

Change: Concentration Determination Paragraph 2

Reason: The preparation and analysis descriptions of the 20%

dose and the reanalysis of the 40% dose on 3/4/94 and 3/5/94

w~e changed to clarify the analyses.

Location: Page ~2

Change: The discussion was changed

Reason: The discussion was changed to clarify the concentration

determination analyses.

study

Kathy

Director: K t-. ,_....-.......---- Staudenmaye%l

Page 18 of 18 !CAN: 902270
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Page 19 of 32

PATHOLOGY REPORT

Compound: Ethylene glycol monobutyl ether

Male and female guinea pigs given 2000. 1000, or 500 mglkg of the test mate:riaJ by gavage,

as part of an acute oral toxicity study. were necropsied. Necropsy lesions are listed in

computer-generated tables.

The cause of death for guinea pigs, which died after exposure to the test mate:rial, was not

determined. However, necrosis and hemOIrhage in the glandular gastric mucosa. may have

contributed to the deaths.

GROSS PATHOLOGY

Male Guinea Pigs - 2000 mglkg dose grouJ): Treatment-related changes in two animals,

which died on Day I, and a single animal, which died on the day of dosing, included minor

to moderate diffuse necrosis (313) and minor hemorrhage (313) in the gastric mucosa and

minor accumulation of mucoid contents in the duodenum (313). The carcasses of two

animals showed minor autolysis. No treatment-related changes were observed in the

remaining two animals that survived the 14-day obselV2.ti.on period.

Male Guinea Pigs - 1000 mg/kg dose groW: Treatment-related changes in a single animal

which died on Day 3 included moderate diffuse necrosis of the gastric mucosa, and minor

red discolOIation of the facial hair. No treatment-related changes were observed in the

remaining four animals which survived the 14-day observation period.

Male Guinea Pigs - 500 mglkg dose grouJ!: No treatment-related changes were observed.

All guinea pigs survived the 14-day observation period. .

Female Guinea Pigs - 2000 roglkg dW.e gI'OlJ1!: Treatment-related changes included minor to

modetate diffuse necrosis (5/5) and minor hemoabage (515) in the gastric mucosa, and minor

accumulaliOD of mucoid contents in the duodenum. (415). Three animals died on the day of

dosing and the remaining two animals died on Day 1. Carcasses of two animals showed

minor autolysis.

Female Guinea Pigs - 1000 mglkg dose group: Treatment-related changes in a single znimal

that died on Day I included minor diffuse necrosis and minima] bemoIIbage in the gastric

i:''JCOS2., and minimal accumulaDon of mucoid contents in the duodenum. No treatmentrdated ch3nges were observed in the remaining four animals, which survived the 14-day

C?bservation period.
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APPENDIX

Page 20 of 32

Female Guinea Pigs - 500 mglkg dose gtOU1!: No tIeatment-related changes were observed.

All guinea pigs survived the 14-day observation period.

All other lesions listed in the attachC:d tables were not considered treatment-related. In the

absence of significant gross organ lesions, other than the obvious signs of gastroin~

iIritation, no tissue was collected for microscopic examination.

Comments: The test material was a gastrointestinal irritant as evidenced by necrosis and

hemonbage in the gastric mucosa and accumulation of mucoid contents in the duodenum.

The cause for incomplete collapse of the lungs upon thoracotomy and lung hemoIIbage that

were observed in Guinea Pigs 176 (1000 mg/kg) and 188 (1000 mglkg) was not determined.

Milan S. Vlaovic, D.V.M., Ph.D.

MSV:dap

04/12194
EXPERIMENT I 9483e9Ae

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APPENDIX

Page 21 of 32

SlMlARY GROSS PA1l1OLOCY INCIDENCE TABLE

COIotPOUNO , 9+-e3ee

SOFTllARE VERS I 3.8

ACCESSION NlU3ER 982270

see.oo9 '009.009 2ee8.ee8

IoIG/I<G IoIG/I<G 1oIG/l<G .. .. ..

TRACHEA 5 5 5

LUNGS 5 5 5

COLI.N'S£ INCOtoFLETE ON l'HORACOTa.rt' 8 , 8

HDICRI'lHf.GE 8 , 8

nmaJS 5 5 5

HEART 5 5 5

• ESOPHAGUS 5 5 5

STCUAOt 5 5 5

IoIJCOSA

NECROSIS. DIFFUSE 8 , 3

HOIJRRHAGE 8 8 3

llUClDOUol 5 5 5

INTESTINAL CONTENTS

INCREASED e e 3

JEJl.INUM 5 5 5.

Il..ELU 5 5 5

C£Cl.U 5 5 5

COLON 5. 5 5

RECTUot 5 5 5

LIVER 5 5 5

KIONEYS 5 5 5

~INAR'r BI,.q)l)ER 5 5 5

PITUITARY GlAND 5 5 5

ADRENAlS 5 5 5

p~. NOS 5 5 5

THYROID GlANDS 5 5 5

PARATHYROID GLANDS S S S

SPLEEN S S 5

MESENTERIC L"BFH NODES S 5 5

BONE MARROW S S S

BRAIN 5 S S

EYES S S 5

SALIVARY GLANDS S S S

ADIPOSE TISSUE S S S

I«IoIBERS REPRESENT7f!ER OF TISSUES EXAMINED. OR IN 1HE CASE OF A8NClRMAL FINDINGS.

THE tu.lBER OF TI Wl~ ~TY -

REVIEWED BY: fkL.W/rP~DATE:~BY~~\1~Wv DATE:tJl,j.fqlj

(J /
CROUP

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APPENDIX

Page 22 of 32

CROSS PATHOLOGY INCIDENCE TABLE

COIFCJUtoD , 94-0386

SOFlWARE VERS' 3.0

ACCESSION ......eER 902278

5e8.ee8 1ee8.ee8 2ee8.ee8

lotC/KC lotC/KC lotC/KC .. M M

SKIN. NOS 5 5 5

HAIR 5 5 5

HAIR OF' FACE

OISCOl..CAATION.RED e e

JroCC£SSOln' SEX ORGANS (WALE) 5 5 5

EPIDIO'tUIDES 5 5 5

TESTES 5 5 5

BODY AS A WHOLE. NOS e 8 2

AUTOl.YSIS e 8 2

ER OF TISSUES EXAMINED. IN THE CASE OF ABNORMA.L FINDINGS.

WIlM EACH ASNORNALITY

REVIEWED B'f:.~!t!l:...::;t.q.L.1i2.l.~DATE: 3::Y)'i"FEPTED B'f~ ~\~~ DATE: ~r~ tj
291l09.D

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APPENDIX

Page 23 of 32

INDIVIDUAL ANI......L CROSS PATHOL.OGY INCIDENCE TABLE

EXPERIMENT , 9~

see.He WGjl<C CROUP - tot

CCU'OUNDl9+-ll3ee SOFlWARE VERS 3.e

AOCESSIOI HUe ge227e

ANINAL ,

DAYS 01 TEST

TRAOtEA

L.l,N;S

TtmIJS

HEART

ESOPHACUS

STOloW:H

OUODENIAI

JEJUNtIoI

ILDM

CECLU

COLON

REC1Uol

UVER

KIDNEYS

URINARY BLADDER

PITUITARY GLAND

AIlR£NALS

PANCftEAS. NOS

THYROID GI.NOS

PARATHYROID GLANDS

SPLEEN

MESENTERIC L'lV'H NOOES

BONE IolARROlI

BRAIN

EYES

SALIVARY GLANDS

ADIPOSE TISSUE

SKIN. NOS

201 282 2e3 2M 285

14 14 14 14 14

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

~R X X X X x

ACCESSORl SEX ORGANS (NALE) X X X X x

EPIDID'lUIDES x x x x X

TESTES X x x x X

KEY:N-NClIlIP.L]iTlSSUE COLLECTED FOR HISTOPAnt:)LOGY. 1-wINlWAL.2-MlNOR.3-VJDERATE.4-SEVERE

P-PRESENT. .~REPORT (FORY 12). X~BUT NOT COLLECTED

R£VIEWED BY: L.+tUJJ&v- DATE: BY~J \\'(.\i\~ DATE:"l{~
291109D

TX-94-96

APPENDIX

Page 24 of 32

INDIVIDUAL ANIMAL CROSS PATHOI..OGY INCIDENCE TABLE

EXPERIMENT , 948388A9 COI'OUND 9+-e3ee SOFTWARE VERS 3.9 ACCESSION toe 982Z78

1998.998 Y:/l<G GROlP - lot

ANIMAL , 176 177 178 179 188

DAYS ON TEST 3 14 14 14 14

lRACH£A . X X X X X

I.J..IoIGS X X X X COLLAPSE INCICIY"LETE ON THORItCOTOIIY P HDO'lRHAGE 2

THIVJS x x x x X

HEART X X X X X

E:SOPItAGUS X X X X X

STCMAOt X X X X

YJOOSA

NECROSIS. DIFFUSE ·3

DUODEM.U X x x x X

JEJtH.N X X X X X

IL.D.U X X X X X

CECUII X X X X X

COLON X X X X X

R£CTtAI X X X X X

LIVER X X X X X

KIDNEYS X X X X X

I.UNARY' BLADDER X X X X X

PIlUITARY" GLAND X X X X X

ADRENA1..S X X X X x

PANCREAS. NOS x X x x X

THrROID CI.NC)S X X X X X

PARATHrROID GI.NOS X x x x x

SPLEEN x x x x X

MES£NTERlC Lnf'H NODES X X X X x

8ClNE IWlRCII X X X X X

BRAIN X X X X x

EYES X X X X X

SALIVARY" GLAfC)S X X X X X

ADIPOSE TISSUE X X X X X

SKIN. NOS X X X lC X

HAIR lC X X X HAIR OF' FACE

DISCOLORATION.RED 2

NX£SSa<f SEX ORGANS (1oIALE) x x x x X

EPIDIlMIlDES X X X X x

TESTES X X X X x

""~"""'1i- .....""""'lHOLClOY.,....-................,..,.TE.......... P-PRESENT. k,...REPOfCT (FORM '2). BUT ND1' c:ou.ECfED

REVlDIED BY: ( lI2 DATE:3~BY:.~J\\\Dau..:.. DATE:'J(oMfj"
EXPERIMENT I 9483e8Ae

291109D

TX-94-96

APPENDIX

Page 25 of 32

INlIVIOUAL ANllIAl. GROSS PA~ IHCIDDCE TABLE

ca.::l 9SOFTWARE VERS 3.0 4-830e

ACCESSION 992278

ANIMAL I

DAYS ON TEST

TlWH:A

I..lH:S

TMYIIJS

HEART

E:SOPtWlUS

S'1'CIMQi

YJCClSA

NECROSIS. DIFFUSE

HaIORRHAGE

.DUODENlIoI

INTESTINAl. CONTDlTS

INCR£\Sm

JE./UNLU

ILEUM

C£D.AoI

CO~

REC1\.U

UVER

KIDNEYS

UUNAR'/' BLADDER

PITUITAR'I" GlAND

ADROW.S

PANCREAS. NOS

lH'tROlD GLNClS

PARAlH'tROID GLNI:lS

SPLEEM

MESENTERIC L'IV'H NODES

ElONE w.RRClW

llRAIN

EYES

SAUVAR'I" CLANDS

ADIPOSE TISSUE

SKIN. NOS

HAIR

ACCESSOR. SEX ORGANS (looIAlE)

EPIDIOYNIDES

TESTES

2888.ee8 1oIG/lCG GRClLP - ..

161 162 163 1~ 165

14 8 14

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X

323

222

X X

222

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X· x x x

x x x x x

x x x x x

x x x x x

x x x x x
291109D

TX-94-96

APPENDIX

Page 26 of 32

Sl.UWtr GROSS PATHOt.OGY' INCIDENCE TABLE

CCIof'OUIC) , 94-e3ee

SOf"TWARE 'WRS' 3.8 N::CESSION tue£R ge2278

:588.888 1eee.888 2eee.eete ~/KC ~/KC 1oC/KC

F F F

TllAQtEA 5 5 5

UN:S 5 5 5

HO«JRRttACE 8 1 e

COUAPSE INCOIoI'LETE ON 11tORACOTOI« e 1 e

TtMIJS 5 5 5

HEART 5 5 5

ESCPKAGUS 5 5 5

STOMACH 5 5 5

YJCOSA

NEalOSIS. DIFFUSE e 5

HBlORRHACE e 5

OUCDEHM 5 5 5

INTESTINAL CQ(I'£NTS

INCR£ASED e 4

JE.IUlI.U 5 5 5

IL£U,I 5 5 5

ca:u. 5 5 5

COLON 5 5 5

RECTLU 5 5 5

UVER 5 5 5

KIDNE\'S 5 5 5

URINARY BLADD£R 5 5 5

PIlUITARY GUM) 5 5 5

AORENALS 5 5 5

PANCRE"AS. NOS 5 5 5

THYROID ClANOS 5 5 5

PARATHYROID CLAtClS 5 5 5

SPLEEN 5 5 5

WESENTERIC L'nFH NODES 5 5 5

BONE MARROW 5 5 5

BRAIN 5 5 5

EYES 5 5 5

SAUVAffY GlANDS 5 5 5

ADIPOSE TISSUE 5 5 S
EXPERIMENT , 9483HA8

291109P

TX-94-96

APPENDIX

Page 27 of 32

SUAlARY GROSS PATHOLOGY' INCIDENCE TA8I..E

COFOlN) , 94-938fI

SOFTWARE VERS' 3.8 ACCESSIQI N.I8ER 982278

5ee.eee 1eee.eee :zeee.eee IIG/ICC IIG/ICC UG/l<G

F F F

SKIN. NOS 5 5 5

HAIR 5 5 5

FALLOPIAN TUBES 5 5 5

VAGINA 5 5 5

UTERUS 5 5 5

HTDRCIoIETRA 8 8 1

OVARIES 5 5 5

CERVIX UTtRI 5 5 5

IIOO'f A WHOLE. NOS e e 2

AUTOLYSIS e e 2
291109.D

TX-94-96

APPENDIX

Page 28 of 32

INDIVIDUAl. ANIMAL GROSS PATHOLOGY INCIDENCE TABLE

:5ee.&ee "'/lCG GRCU' - F

2e6 267 268 269 216

\4 14 14 14 14

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

ANIMAl. ,

DAYS ON TEST

TRAa1EA

u.t«;S

'IlM&JS

HEART

ESOPHAGUS

STOoIAOt

DUQDEJU,I

.IEJUNM

ILEUI

CEQM

COLON

REC1\.IoI

UVER

KIDNEYS

UUNARY BlADOER

PITUITARY GU\ND

ADRaW.S

PANCREAS. NOS

THYROID CL»I)S

PARATHYROID CI.»lDS

SPLEEN

IoIESOfrERIC L'M'H NODES

BONE MARROI

tlRAlN

EYES

SALIVARY ClNlDS

ADIPOSE TISSUE

SKIN. NOS

HAIR

FALlOPIAN TUBES

VAGINA

UnRUS

x x x

x x x

x x x

x x x

x x x

x X x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x x

x x

x x

x x

x x

x x

x x

x x

x x

x X

X X

X X

X X

X X

X X

X X

X X

X X

X X

X X

X X

X X

X X

X X

X X

X X

aM"CUI)' SOFTWARE VERS ,- 3.6

ACCESSION tUeER 962276

OVARIES X X X X X

CERVIX UTERI X X X X X

ICEY:N-HClRIM.L.]lISSUE COLLECTED FOR HISTOPAntDl..OG'l".1-YINIW.l..:HIlNOR.3-UODERATE.+-SEVERE

p...flftESE)(T. • REPORT (f'QRW '2). x-NORMAI. BUT NOT COLLECTED •

REVIEWED B'f:~ ?ttrlf?1...&. DATE:~ BY~"t \!q~U,.... DAT£:"Iro/qr/
291109D

TX-94-96

APPENDIX

P~e29 of 32

ItI>IVIOUAL AMINAL CROSS PATt1Ol..OG'r' INCIDENCE TABLE

aM'C'.N)l M-e30e SOFTWARE VERS 3.8 I'CCESSICI'l NI..IoS 992270

'_.808 MG/kG GRCJU> - F

186 187 188 189 198

14 , ... 14 14

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

X X X X X

ANII.W. I

DA'YS CI'l TEST

TRACHEA

LUNGS

COUAPSE:

H~

INCaoFLETE ON THORACOTOWY

1HnAJS

HEART

ESOPHAGUS

S'TCINACH

I&JOOSA

NB3)SIS. DIFFUSE

HBIORRHAGE

-DUClDEJ«AA

INTESTINAL CONTENTS

INCREASED

JEJlH.U

ILEIM

CECUoI

COLON

RECTUoI

UVER

KIDNEYS

lIUNARY BlMJOER

PITUITARY GlN«)

AIlRENA1.S

PNCREAS. NOS

THYROID Cl.AN)S

PARATHYROID GLANDS

SPl.EDl

MESENTERIC L'nAt NODES

BONE UARRCW

ElRAlN

EYES

SALIVARY GlN«)S

ADIPOSE TISSUE

SKIN. NOS

HAIR

FALLOP~ TUBES

VAGINA

UT£RUS

OVARIES

CERVIX UTER1

x

X

X

X

X

X

X

X

X

x

X

X

X

X

X

...

P

X

X

X

2

1

X

X

x

X

X

X

X

X

X

X

X

X

X

X
2911000

TX-94-96

APPENDIX

Page 30 of 32

INDIVIIlUAI. ANIUAl. CROSS PAntOl..OCOl' INCIDOICE TABLE

EXPERIWENT I 94e380A8

SOFTWARE~~VERS 3.8

ACCESSION 992278

2e8e.lIe8 UC/KG CROU" - F

ANIUAl. I 166 167 168 169 178

DAYS ON TEST e e 8

TRAOIEA X X X X X

lam X X X X X

llmAJS X X X X X

HEART X X X X X

ESClPIWiUS X X X X X

STaw::H

IofJCOSA

N£CROSIS. DIFFUSE 3 2 3 2 2

HDI:lRRHAGE 2 2 2 2 2

-OUCOEt«N X

INTESTINAl. 00NTDlTS

INCREASED 2 2 2 2

JE.nI«AI x X x x X

ILEIIoI X X X X X

cm.u x x x x x

CQltIol x x x x X

RECTI.IIIl X X X X X

LIVER X X X X X

KIDNEYS X X X X X

LRlNAR't' BLADDER X X X X X

PIlUITAR't GLAND X X X X x

ADRDW.S X X X X X

PNCREAS. NOS x X x x X

lHYROlO CI.AMlS X X X X X

PARATHYROID GLANDS X X X X X

SPL£EH X X X X X

IoIESEN1'ERIC LYIAI NODES x x x x x

BONE MARROIf X X X X X

BRAIN X X X X X

EYES X X X X X

SALIVAR't ClNlDS X X X X X

ADIPOSE T1ssuE x x x x X

SKIN. NOS X X X X X

HAIR X X X X X

FALLOPIAN TUBES X X X X X

VAGINA X X X X X

UTERUS X X X x

H'I'DRClMEIRA 2

OVARIES x x x x x

CERVIX UI'ERI X X X X X

ICEY:N-NORIML TISSUE coua:::rm FOR HISTClPATHOLClGY.'I-MINDIAL.2-UINCR.3-4d)ERA.TE.+-SEVERE

"'--;:;J;"!Jzl~ ,..... Pl. - "" .... """""'" .•

REVIEJIED 8'1": :' ~~11P DATE: 3·3?J~ACCEPTED 8'1"~..& \J \ ~U- DATE: '1fc:-'9t/
2911~D

TX-94-96

APPENDIX

Page 31 of 32

UOIVIDUAL. ANIWA!. CROSS PATHOLOGY INCIDENCE TABLE

SOFTWAREau:::l~ vms 3.8

ACCESSION 982278

zeee.ee&- WG/lCG CROl.P - F"

ANIlW. , 166 167 168 169 178

DAYS (IN TEST 8 8 8

800Y IS A WHOLE. NOS

AUTOLYSIS 2 2

1CEI':~ISSlJE COLLECTm FaR HISTClPA1ltOLOGY.11lINIWA!..2-UINOR.3-IoIXlERATE.4-SEVERE

:-4 iCESWT. .-~~ REPORT (~ '2). BUT NOT COLLECTED

REVlEJIED BY: .~{'- 'l!(pJ.- DATE: 3·~ BY:~A \f ,~u.:...o....TE:'-Ir(.Hqy

f
EXPERIMENT , 9483lKWl

291109.0

T-X-94-96

APPENDIX

Page 32 of 32

GROSS PATHOLO(;'( CClI&I£NT REPORT

COWPOUND , 94--03ee

SOFTWARE VERS' ;S.B

ACCESSION MMlER 982278

DAY DOSE LEVEL AHIWAL-' ---- --- 2.3 18ee.8ee AGjKG 188

2.3 2eee.8ee AG/lCG 162

23 2eee.8ee AGjKG 164

2.3 2eee.ee8 AG/KG 166

23 2888.8ee AG/lCG 168

2.3 2eee.8ee AGjKG 163

2.3 2eee.8ee IIG/KG 167

CCII.IENT

OUOOBUI HAD UJCOID eotn'ENTS.

OUOOBUI HAD UJCOID CQolTENTS.

OUOOBUI HAD UJCOID CQolTENTS.

OUOOENlM HAD UJCOID CONTENTS.

OUOOBUI HAD UJCOID eotn'ENTS.

OUODENUoI HAD UJCOID eotn'ENTS.

DUODBUI HAD IoIJCOIO eotn'ENTS.

23 2eee.8ee AG/KG 170 DUODBUI HAt' IoIJCOID eotn'ENTS.

REVIElIED B'I': ft~p~ DATE:3-3:>-1'fKUPTm SY:~~V~~~DATE:'I(C'1V

p