# REVISED FINAL REPORT

AN ORAL REPRODUCTION/DE VELOPMENTAL TOXICITY

SCREENING STUDY OF ANTHRAQUINO NE (CAS # 84-65-1) IN RATS

TEST ARTICLE: Anthraquinone (CAS # 84-65-1)

TESTING FACILITY: MPI Research, Inc.

54943 North Main Street

Mattawan, Michigan 49071-9399

STUDY NUMBER: 1271-001

STUDY DIRECTOR: Cheryl K. Lanphear, B.S.

SPONSOR: Chemical Products Corporation

102 Old Mill Road, SE

P.O. Box 2470

Cartersville, Georgia 30120-1692

SPONSOR REPRESENTATIVE: Jerry A. Cook

DATE OF STUDY COMPLETION: February 6, 2007

DATE OF REVISED FINAL REPORT: May 4, 2012

Volume 1 of 1 Page 1 of 358

# 1271-001 REASON FOR REVISED FINAL REPORT

This report is a revised version of the origin al final report of MPI Research Study Number

1271-001, titled “An Oral Reproduction/Develo pmental Toxicity Screening Study of

Anthraquinone (CAS # 84-65-1) in Rats,” issued on February 6, 2007. The following

revisions have been made. This is a new report. No changes to the raw data were made.

1. Section 2. Summary – Added discussion regarding the actual measured dose

levels administered based on analysis results, and discussion regarding

postimplantation loss.

2. Section 2. Summary – Revised concluding sentence to indicate that maternal

NOEL was also not achieved.

3. Section 4.5. Statistics – % Postimplan tation Loss and Arcsin-Square-Root

Transformation was added to the statistical comparisons.

4. Section 5.1.1. Homogeneity – Added “Average Measured Dose Level

(mg/kg/day)” to in-text summary tables.

5. Section 5.1.3. Concentration – Added disc ussion regarding the actual measured

dose levels administered based on analysis results and “Average Measured Dose

Level (mg/kg/day)” to in-text summary tables.

6. Section 5.5. P Delivery and F 1 

# Litter Data – Added discussion regarding

postimplantation loss.

7. Section 6. Conclusion – Revised to include maternal NOEL not achieved at the

dose levels evaluated, and other effects seen at parturition (i.e., decrease in pups

born, fewer liveborn, etc.).

8. Reproduction Glossary – Added calculation for % Postimplantation Loss.

9. Table 23. Summary of P Natural Deli very and Litter Data – Added

% Postimplantation Loss.

10. Appendix B. Section 2 Analysis of Dosi ng Formulations – Average Dose Level

(mg/kg/day) calculated for homogeneity and concentration.

11. Appendix N. Individual P Natural Deliv ery and Litter Data – Added individual

calculated % Postimplantation Loss.

12. Appendix R. Protocol and Amendments – Added Protocol Amendment 4 issued

on March 27, 2012. Page 2 of 358

# Page 3 of 358

# Page 4 of 358

# 1271-001 MPI RESEARCH KEY PERSONNEL

Chairman & Owner, CEO William U. Parfet, M.B.A.

President, Chief Operating Officer William M. Harrison, B.S.

Executive Vice President,

Director of Research James Laveglia, Ph.D.

Study Director Cheryl K. Lanphear, B.S.

Alternate Contact Raymond E. Schroeder, M.S., D.A.B.T. ®



# Senior Director, Laboratory

Operations Benjie A. Culp, A.L.A.T.

Anthony Rohr, B.S., A.L.A.T.

Project Leader, Toxicology Services

Director, Clinical Laboratory

Medicine Allan G. Manus, D.V.M., M.S., D.A.C.L.A.M.

Director, Pathology Theodore W. Slone, Jr., D.V.M., D.A.C.V.P.

Senior Principal Pathologist

Dennis T. Nelson, D.V.M., M.S., D.A.C.V.P.

Study Pathologist

Senior Director, Regulatory Affairs Mary Ann Scott, B.S.

Report Services Christy E. Kubin, B.S.

Manager, Report Services

Brenda L. Gaza

Report Coordinator I, Report Services

Quality Assurance Katherine Longman, B.S., RQAP-GLP

Manager, Quality Assurance Research

Kelle Brooks, M.A., RQAP-GLP

Senior Quality Assurance Auditor Page 5 of 358

# 1271-001 TABLE OF CONTENTS

Page

REASON FOR REVISED FINAL REPORT............................................................................ 2

STATEMENT OF COMPLIANCE........................................................................................... 3

SIGNATURE......................................................................................................................

....... 4

MPI RESEARCH KEY PERSONNEL ..................................................................................... 5

1. QUALITY ASSURANCE STATEMENT ........................................................................ 9

2. SUMMARY..................................................................................................................... 10

3. INTRODUCTION ........................................................................................................... 12

3.1. Objective.................................................................................................................

.. 12

3.2. Species Selection ...................................................................................................... 12

3.3. Justification for Number on Study............................................................................ 12

3.4. Study Schedule.......................................................................................................... 13

4. MATERIALS AND METHODS..................................................................................... 14

4.1. Vehicle and Test Ar ticle Information ....................................................................... 14

4.2. Experimental Design................................................................................................. 16

4.3. In-life Examinations.................................................................................................. 17

4.4. Postmortem Study Evaluations................................................................................. 19

4.5. Statistics ................................................................................................................

.... 20

4.6. Computer Systems .................................................................................................... 23

4.7. Data and Specimen Retention................................................................................... 23

5. RESULTS AND DISCUSSION...................................................................................... 24

5.1. Analysis of Dosing Formulations ............................................................................. 24

5.2. In-life Examinations.................................................................................................. 27

5.3. Postmortem Study Evaluations................................................................................. 28

5.4. Reproductive Performance........................................................................................ 29

5.5. P Delivery and F 1 

# Litter Data.................................................................................... 30

5.6. F 1 

# Pup Evaluations .................................................................................................... 30

6. CONCLUSION................................................................................................................ 32

REPRODUCTION GLOSSARY.............................................................................................33

PATHOLOGY GLOSSARY...................................................................................................36

Table

1. Summary of P Clinic al Findings - MALE .........................................................................38

2. Summary of P Premating/Mating Clinical Findings – FEMALE......................................41

3. Summary of P Gestatio n Clinical Findings .......................................................................44

4. Summary of P Lactati on Clinical Findings........................................................................47

5. Summary of P Body Wei ght Values - MALE ...................................................................50

6. Summary of P Premating Body Weight Values – FEMALE ............................................52

7. Summary of P Gestatio n Body Weight Values..................................................................54

8. Summary of P Lactati on Body Weight Values..................................................................56 Page 6 of 358

# 1271-001 TABLE OF CONTENTS

Page

Table (Cont.)

9. Summary of P Gestation B ody Weight Change Values ....................................................58

10. Summary of P Lactation B ody Weight Change Values.....................................................60

11. Summary of P Food Consumption Values – MALE .........................................................62

12. Summary of P Premating Food Consumption Values – FEMALE ...................................64

13. Summary of P Gestation Food Consumption Values ........................................................66

14. Summary of P Lactation Food Consumption Values ........................................................68

15. Summary of P Macroscopic Observations – MALE .........................................................70

16. Summary of P Macroscopic Observations – FEMALE.....................................................72

17. Summary of P Organ We ight Values – MALE .................................................................75

18. Summary of P Organ Wei ght Values – FEMALE.............................................................77

19. Summary of P Microscopi c Observations – MALE..........................................................79

20. Summary of P Microscopic Observations – FEMALE .....................................................81

21. Summary of Premating Estrous Cycling............................................................................83

22. Summary of Reproductive and Fertility Parameters..........................................................85

23. Summary of P Natural Delivery and Litter Data ...............................................................88

24. Summary of F 1 

# Pup Clinical Findings................................................................................92

25. Summary of F 1 

# Pup Body Weight Values..........................................................................94

26. Summary of F 1 

# Pup Macroscopic Observations ................................................................96

Appendix

A. Vehicle and Test Ar ticle Information..............................................................................103

B. Analytical Report.............................................................................................................1 06

C. Record of Animal Fate and Disposition ..........................................................................128

D. Individual P Clinical Findings.........................................................................................133

E. Individual P Body Weight Values ...................................................................................159

F. Individual P Body Weight Change Values ......................................................................170

G. Individual P Food Consumption Values..........................................................................176

H. Pathology Interpretation ..................................................................................................186

I. Tissues and Organs Collected, Weighed, and Examined ................................................189

J. Individual P Orga n Weight Values..................................................................................191

K. Individual P Animal Data Record: Pathology ................................................................205

L. Individual P Premating/Ma ting Estrous Cycling Data.....................................................229

M. Individual P Mating Record............................................................................................237

N. Individual P Natural Delivery and Litter Data ................................................................240

O. Individual F 1 

# Pup Clinical Findings.................................................................................245 Page 7 of 358

# 1271-001 TABLE OF CONTENTS

Page

Appendix (Cont.)

P. Individual F 1 

# Pup Body Weight, Sex, and Status Data ....................................................275

Q. Individual F 1 

# Pup Animal Data Record: Pathology........................................................283

R. Protocol and Amendments...............................................................................................332

S. Deviations ..................................................................................................................... ...357 Page 8 of 358

# Page 9 of 358

# 1271-001 2. SUMMARY

This study was conducted for Chemical Products Co rporation to evaluate the effects of the

test article, Anthraquinone (CAS # 84-65-1), on male and female reproductive performance,

including gonadal function, estr ous cycle, mating behavior, c onception, development of the

conceptus, and parturition in rats. Three treatment groups of 12 male and 12 female

CD ® 

# [Crl:CD ® 

# (SD)] rats were administered the test article at intended dose levels of 150,

600, and 2400 mg/kg/day, at a dose volume of 20 mL/kg. One additional group of 12

animals/sex served as the control and received th e vehicle, 1% Guar Gum in distilled water.

The vehicle or test article was administered via oral gavage once daily to the males and

females beginning 14 days prior to mating. Dosing of the males continued through the

mating and postmating period to euthanasia wh ile dosing of the females continued through

the mating and gestation periods to Lactation Day (LD) 4.

Observations of the animals included clinical signs, body weights, and food consumption.

After at least 14 days of treatment, the anim als were cohabited, one male and one female

within the same treatment group for up to 14 da ys. Females were allowed to deliver litters

and maintain the pups until LD 4. Litters were evaluated for the number of live born or

stillborn pups, pup survival to LD 4, individu al pup body weights and sex, and pup external

gross abnormalities on LD 0 and 4. At study termination, necropsy examinations were

performed on the adult P animals, and organ we ights were recorded. Selected tissues were

microscopically examined for animals at 0 a nd 2400 mg/kg/day. Pups were euthanized and

externally examined on LD 4, and the carcasses were discarded.

Preparation of the dosing suspen sions was problematic. Due to the viscosity of the vehicle

(1% Guar Gum), the formulations were unable to be stirred adequatel y, and therefore it was

difficult to obtain uniform suspensions. Periodi c concentration analysis of the test article

suspensions prepared for study determined that the formulations for the most part were

neither homogenous nor were the intended concen tration levels achieve d. Recoveries ranged

from approximately 60-80% of nominal in most instances, except during Week 8 when

recovery at the low concentration was 25.1% of nominal and the high concentration was

99.5%. Based on these recoveries the actua l measured dose levels were 109, 396, and

1661 mg/kg/day for Week 1, 116, 357, and 2034 mg/kg/day for Week 3, and 38, 463, and

2388 mg/kg/day for Week 8, respectively, fo r the 150, 600, and 2400 mg/kg/day dose levels.

Test article-related mortality was seen in fe male rats at 600 and 2400 mg/kg/day. Eight and

seven females died or were euthanized in extremis in the 600 and 2400 mg/kg/day group,

respectively. Most of this mortality occurred during gestation and lactation. One male in the

150 mg/kg/day group died, but in the absence of mortality among males at the higher dose

levels and since macroscopic findings were unremarkable, this death was considered

unrelated to treatmen t with Anthraquinone.

Test article-related macroscopic changes we re noted in female rats that received

Anthraquinone, but not in males. Black or re d foci were observed in the glandular stomach

of females at 600 and 2400 mg/kg/day that died and an erosion/ulcer wa s noted in the pyloric

stomach of one female that died at 2400 mg /kg/day. Also at 2400 mg/kg/day, minimal focal Page 10 of 358

# 1271-001 necrosis and moderate periportal vacuolation of hepatocytes were observed in one female,

edema was seen in two of seven animals that di ed, and erosion/ulcer in the glandular stomach

was seen in three of these animals.

At all dose levels in both males and females treatment related toxicity was observed and

consisted of discolored urine (brown, orange , or red), lower body weights and body weight

gain, and lower food consumption. No effect of treatment was eviden t from mating, fertility,

and fecundity indices but treatment did pr oduce a lengthening of the estrous cycle.

The pregnancy index was 91.7, 83.3, 100, and 91.7% in the control, 150, 600, and

2400 mg/kg/day groups, respectively. All pregnant animals in the control and

150 mg/kg/day groups delivered and retained litters through LD 4. In the 600 mg/kg/day

group, 10 females delivered, but only two female s retained litters through LD 4. In the

2400 mg/kg/day group, four female s delivered, but only one reta ined a litter through LD 4.

The number of pups delivered (live + dead) was significantly decreased in the 600 and

2400 mg/kg/day groups. The number of liveborn pups in these groups was decreased with an

increase in stillborn pups. Additionally, the m ean number of total implantation scars/litter

was significantly lower in these groups a nd the mean post-implan tation loss was higher

compared to controls. These effects (i.e., de crease in mean number of total implantation

scars, fewer number of pups de livered (live plus stillborn), an d increase in post-implantation

loss) at 600 and 2400 mg/kg/day were considered te st article-related. Pu p survival to LD 4,

although not dose-dependent, for the 600 and 2400 mg/kg/day groups wa s significantly lower

(16.5 and 33.3%, respectively) than control. At 150 mg/kg/day, no effect of treatment was

evident from parturition data, litt er size, or pup survival to LD 4.

Mean pup body weights (by sex and sexes co mbined) in the 150 mg/kg/day group were

similar to control at birth (LD 0), but 8-10% lower by LD 4. In the 600 and 2400 mg/kg/day

groups, pup weights at birth and LD 4 were not ably lower than cont rol. The lower pup

weights at all dose levels were considered indicative of a treatment-related response.

Based on these results, neither a paternal, maternal, nor a developmental No-ObservedEffect-Level (NOEL) for Anthraquinone was achieved. Page 11 of 358

# 1271-001 3. INTRODUCTION

This study was conducted in accordance with Sta ndard Operating Procedures (SOP) and the

protocol as approved by the Sponsor. This study was based on Guideline 421,

Reproduction/Developmental Toxicity Screen ing Test, the Organization of Economic

Cooperation and Development (OECD) Guide line for Testing of Chemicals, revised

July 1995. The protocol and amendments are pres ented in Appendix R. Procedures pertinent

to this study are described in this report.

3.1. Objective

The objective of this study was to evaluate the ef fects of the test article on male and female

reproductive performance, including gonadal function, estrous cycle, mating behavior,

conception, development of the concep tus, and parturition in rats.

3.2. Species Selection

The current state of scientific knowledge a nd the applicable guidelines do not provide

acceptable alternatives in vitro or otherwise to the use of live animals to accomplish the

purpose of this study. “The development of kno wledge necessary for the improvement of the

health and well-being of humans as well as other animals requires in vivo experimentation

with a wide variety of animal species”. 1 

# “Whole animals are essential in research and testing

because they best reflect the dynamic interac tions between the various cells, tissues, and

organs comprising the human body”. 2 

# The rat is the usual rodent model 3 

# used for evaluation

the toxicity of various classes of chemicals and for which there is a larg e historical database.

3.3. Justification for Number on Study

This study was designed to use th e fewest number of animals po ssible, consistent with the

objective of the study, the scientific needs of th e Sponsor, contemporary sc ientific standards,

and in consideration of applicable re gulatory requirements cited previously.

1 

# “Principles for the Utilization and Care of Vertebrate Animals Used in Testing, Research, and Training”,

Federal Register, Vol. 50, No. 97, May 20, 1985

2 

# “Position Statement on the Use of Animals in Research”, NIH Guide 22: No. 8, Feb.26, 1993

3 

# Guidance for Industry, Investigators, and Reviewers: Exploratory IND Studies, U.S. F.D.A. Center for Drug

Evaluation and Research (CDER). January 2006. 

# Page 12 of 358

# 1271-001 3.4. Study Schedule

Protocol Approved by Sponsor June 12, 2006

Study Initiation Date (Protocol Signed by Study Director) June 12, 2006

Experimental Starting Date (receip t of test system) June 14, 2006

Experimental Start Date June 28, 2006

Experimental Termination Date August 21, 2006

Experimental Completion Date (Final Data Collection) September 14, 2006

Draft Report Mail Date November 3, 2006 Page 13 of 358

# 1271-001 4. MATERIALS AND METHODS

4.1. Vehicle and Test Article Information

Pertinent vehicle and test article receipt information is presented in Appendix A.

Documentation of the strength, purity, compositi on, stability, and other pertinent information

for the batch of vehicle used on study was limited to that information listed on the label of

this commercially available product. The Sponsor has pr ovided documentation of the

strength, purity, composition, stability, and other pertinent information for the lot of test

article used on study.

4.1.1. Vehicle and Test Article Preparation

To prepare the vehicle, 1% Guar Gum in di stilled water, the requi red amount of powdered

Guar Gum was weighed and slowly dispersed in the required am ount of distilled water using

a motor-driven propeller. The contents of th e beaker were stirred until dissolved. The

solution was transferred in portions into a grad uated cylinder. The cylinder was inverted and

the contents mixed thoroughly betw een additions, and the contents were transferred into an

appropriately sized container. The mixing contai ner was rinsed with distilled water, and the

rinse was transferred into the cylinder. Di stilled water was added to the cylinder, the

cylinder was inverted and the contents mixed thoroughly, and transferred into the container

to yield the required volume of prepared vehi cle. The contents of the container were

thoroughly mixed using a motor-driven propeller.

Fresh vehicle was prepared for use on study week ly, prior to handling th e test article, and

was stored refrigerated. To dispense the vehicle for dosi ng, the required volume of prepared

vehicle was measured using a graduated cylinder and transferred into a beaker. The contents

of the beaker were stirred using a motor-driven propeller. The contents of the beaker were

evenly dispensed into amber gl ass containers using a syringe, while stirring. On occasion,

additional preparations were made as necessary during the course of the study.

The test article was used as received from the Sponsor and no adjustment was made for

purity. To prepare the test ar ticle formulations at each conc entration, the required amount of

prepared vehicle was measured using a graduated cyli nder and transferred into a beaker. The

required amount of test articl e, Anthraquinone (CAS # 84-65-1), was weighed and slowly

transferred into the beaker containing the vehicle while mixing with a Silverson

homogenizer. The weighing vessel was rinsed with vehicle, and the rinse was transferred

into the beaker while mixing. The contents of the beaker were mixed using the Silverson

homogenizer until uniform in appearance. The c ontents of the beaker were transferred into a

graduated cylinder. The beaker and the Silverso n head were rinsed with vehicle and the rinse

was transferred into the cylinde r. Vehicle was added to the cylinder to yield the required

volume of prepared test article formulation. The cylinder was thoroughly inverted and the

contents thoroughly mixed, and transferred into a beaker. The contents of the beaker were

stirred using a motor-driven propeller and even ly dispensed into amber glass containers,

while stirring, using a syringe. Page 14 of 358

# 1271-001 Formulations of the test articl e were prepared for each concentr ation weekly, and were stored

refrigerated. On occasion, additional preparatio ns were made as necessary during the course

of the study.

4.1.2. Analysis of Dosing Formulations

4.1.2.1. Homogeneity

Prior to initiation of test ar ticle administration, from each test batch, and from the third

preparation, duplicate samples (2.0 mL) of the dosing formulations at the 150 and 2400

mg/kg/day dose levels were collected to assess the homogeneity of the test article

preparations employing the same method and ba tch size used during the study. The samples

were collected from the top, middle, and bottom of the container, while stirring, using a

syringe, and placed into 20 mL scintillation vi als. Following completion of the study, any

remaining samples were properly disposed.

4.1.2.2. Stability

Samples (2.0 mL) of the prepared formulati ons mixed for the homogeneity analysis were

stored under refrigerated conditions for up to 14 days. Samples of these stored preparations

were collected after 7 and 14 days at refrigerated temperature to determine the stability of the

test article in the vehicle. Following comple tion of the study, any remaining samples were

properly disposed.

4.1.2.3. Concentration

Samples (2.0 mL) of the dosing formulations at each concentration were collected from

Weeks 1, 3, and 8 to assess the concentration of th e test article. The samples were collected

from the middle of the container, while sti rring, using a syringe, and placed into 20 mL

scintillation vials. Followi ng completion of the study, any re maining samples were properly

disposed.

4.1.2.4. Analyses

All samples were stored refrigerated until an alyzed. All analytical work was conducted by

MPI Research, Inc., using an analytical me thod developed by MPI Research, Inc. and

validated under MPI Research Study Number 1271-002.

4.1.3. Reserve Sample and Test Article Disposition

A reserve sample from the lot of test article us ed in this study was co llected and archived.

Any remaining test article will be shipped to a Sponsor-designated location after completion

of the study. Page 15 of 358

# 1271-001 4.2. Experimental Design

4.2.1. Animal Acquisition and Acclimation

A total of 58 male and 58 female rats [Crl:CD ® 

# (SD)], approximately 6 weeks of age, were

received from Charles River Laboratories, Portage, Michigan on June 14, 2006. During the

2 week acclimation period, the animals were we ighed and observed twi ce daily with respect

to general health and any signs of disease. All animals we re given a detailed clinical

examination prior to selection for study. The pretest body weight s and clinical examinations

performed during the acclimation period are not reported, but are ma intained in the study file.

4.2.2. Randomization, Assignment to Study, and Maintenance

All animals assigned to study had body weights that fell within ± 20% of the mean body

weight for each sex. Animals considered suit able for study were weighed prior to treatment

and randomized, by sex, into treatment gr oups using a standard, by weight, block

randomization procedure. Extra rats obtained for the study, but not used, were euthanized

via carbon dioxide inha lation and discarded.

Forty eight male rats (weighing 237 to 263 gr ams at randomization) and 48 female rats

(weighing 186 to 208 grams at randomization) we re assigned to the control and treatment

groups identified in the table below.

Group Assignments

Number of Animals Group

Number Dose Level

(mg/kg/day) Male Female

1 0 12 12

2 150 12 12

3 600 12 12

4 2400 12 12

Each animal was assigned an animal number to be used in the Provantis TM 

# data collection

system and was implanted with a microchip bearing a unique identi fication number.

The individual animal number, implant num ber, and study number comprised a unique

identification for each animal. Each cage was identified by the animal number, study

number, group number, and sex. Pups were identified by tattoo on LD 0. Animal

identification was verified dur ing the course of the study, as documented in the data.

Animals found dead during the firs t week of the study were replaced. The cause of death for

these animals was determined to be gavage error (perforation of the thoracic cavity). A total

of 11 rats were utilized as replacement animal s and were assigned a unique animal number.

The data recorded for the replaced animals ar e not reported, but are maintained in the study

file.

Throughout the study, all animals were kept in an environmentally controlled room.

Fluorescent lighting was provide d for approximately 12 hours per day. Temperature and

humidity were continuously monitored and reco rded. The protocol-des ignated ranges were Page 16 of 358

# 1271-001 64 to 79° F and 30 to 70%, respectively. The actual temperature and humidity findings are

not reported, but are maintain ed in the study file. Animal s were individually housed in

suspended, stainless steel, wire-mesh type cag es, except during pairing, near parturition, and

during lactation. During pairi ng, animals were cohabited, one male and one female within

the same treatment group. On approximate ly Gestation Day (GD) 20, females were

individually housed in plastic cages containing wood chip beddi ng. Females were housed in

these cages for the remainder of gestation and with their litters through LD 4.

Lab Diet ® 

# (Certified Rodent Diet #5002, PMI Nutri tion International, Inc.) was available

ad libitum

. 

# The lot number from each diet lot used fo r this study was recorded. Certification

analysis of each diet lot was performed by the manufacturer. Tap water was available

ad libitum via an automatic watering system. The water supply is monitored for specified

contaminants at periodic interval s according to SOP. The resu lts of food and water analyses

are retained in the Archives. The Study Direct or is not aware of any potential contaminants

likely to be present in the diet or water that would have interfered w ith the results of the

study. Therefore, no analyses other th an those stated above were conducted.

4.2.3. Test Article Administration

4.2.3.1. Justification for Route of Administration

The oral route is one of the potential routes of human e xposure to this test article.

4.2.3.2. Justification of Dose Levels

The dose levels were selected by the Sponsor, or in consultation with the Sponsor, on the

basis of available data from previous studies.

4.2.3.3. Administration

Vehicle and test article admini stration began at least 14 days prior to pairing and continued

until euthanasia for the males, and at least 14 days prior to pairing through LD 4 for the

females. The test article was administered to the treated groups once per day at the intended

dose levels of 150, 600, and 2400 mg/kg/day, at a dose volume of 20 mL/kg. The control

animals received the vehicle, 1% Guar Gum in distilled water, at the same volume and

dosing regimen as the treated groups. Dosing wa s performed at approximately the same time

each day (± 2 hours from Day 1 start of dosing) . Individual doses we re based on the most

recent body weights.

The control and test articles were administered via oral gavage using an appropriately sized

syringe attached to a Fuchigami ® 

# dosing needle. The dosing formulations were shaken

vigorously between each animal to ensure the dosing formulation was homogenous. The

dosing apparatus was wiped free of te st article prior to administration.

4.3. In-life Examinations

4.3.1. Cageside Observations

All animals were observed for morbidity, mortal ity, injury, and the ava ilability of food and

water twice daily throughout the duration of th e study. On occasion, veterinary consultations Page 17 of 358

# 1271-001 were conducted during the course of the st udy. All treatments and observations were

recorded. The medical treatments and observati ons are not reported, but are maintained in

the study file.

4.3.2. Detailed Clinical Observations

Daily during treatment (approximately 30 to 90 minutes postdose), each animal was removed

from the cage and given a detailed clinical ex amination. The observations included, but were

not limited to, evaluation of th e skin, fur, eyes, ears, nose, oral cavity, thorax, abdomen,

external genitalia, limbs and feet, as well as eval uation of respiration.

4.3.3. Body Weights and Body Weight Changes

Individual body weights for the males and female s were recorded the da y after arri val, prior

to randomization, at initiation of test article administrati on, weekly during the premating

period, and weekly for males and umated females. All mated females were also weighed on

GD 0, 7, 14, and 20, and on LD 0 and 4. Body wei ght change for the females was calculated

for the following intervals: GD 0-7, 7-14, 14-20, 0-20, and LD 0-4. All body weights

recorded prior to the initiati on of test article administra tion are not reported, but are

maintained in the study data.

4.3.4. Food Consumption

Food consumption for all rats was measured a nd recorded weekly pr ior to pairing. Food

consumption was not recorded during the mating period because the anim als were cohabited.

Following the cohabitation period, food consump tion was recorded weekly for males and

unmated females until euthanasia. Food consum ption for the mated females was measured

and recorded on GD 0, 7, 14, and 20, and LD 0 and 4.

4.3.5. Estrous Cyclicity

At initiation of test article administration and until evidence of copulation was observed, or

the cohabitation period ended, the females were examined daily by vaginal lavage to

establish estrous cyclicity. Vaginal lavage of the females was completed daily by

10:00 A.M.

4.3.6. Breeding Procedures

After 2 weeks of treatment, the males were ra ndomly cohabited, one male to one female,

from the corresponding treatment or control group. Each female was housed in the cage of a

male during pairing. Positive evidence of copul ation was established by daily inspection for

a copulatory plug in the vagina and/or sperm present in th e vaginal lavage. The day on

which positive evidence of copulation was obser ved was considered GD 0. After evidence of

mating was observed, the female was returned to an individual cage. The maximum mating

period was 14 days, at the end of which any females with no confirmed evidence of mating

were returned to individual cag es until scheduled euthanasia. Page 18 of 358

# 1271-001 After the mating period, the males were indivi dually housed and continued on treatment for

approximately 2 weeks. At the end of this pe riod, the males were euthanized and necropsied

as described in Section 4.4. Postmortem Study Evaluations.

4.3.7. P Parturition and F 1 

# Litter Observations

Toward the end of the gestation period, female s were examined twice daily for signs of

parturition. The mated females were allowed to give birth (F 1 

# ). The duration of gestation

was calculated, and any difficulties occurring at parturition were recorded. The day on which

all pups were delivered was designated as LD 0. The litters were examined as soon as

possible after delivery and para meters including litter size, the sex of the pups, number of

dead and live born pups, and any gross abnormalitie s of the pups were recorded. Any intact

pups found dead at birth were externally exam ined, stillborn status was determined by lung

floatation in water, and the carcasses were discarded. Any intact pups found dead during

lactation were externally examined and discarded.

Pups were weighed, sexed, and examined for external abnormalities on LD 0 and 4. On

LD 4, the pups were euthanized via intraper itoneal injection of sodium pentobarbital

solution, externally examined and any abnormaliti es were recorded, and the carcasses were

discarded.

4.4. Postmortem Study Evaluations

4.4.1. Macroscopic

Necropsy examinations were performed unde r procedures approved by a veterinary

pathologist on all adult animals euthanized in extremis, found dead, and all surviving animals

at the scheduled necropsy. Euthanasia was by carbon dioxide inhalation, followed by

exsanguination via abdominal vena cava.

The animals were examined carefully for extern al abnormalities including masses. The skin

was reflected from a ventral midline incisi on and any abnormalities were identified and

correlated with antemortem findings. The a bdominal, thoracic, and cranial cavities were

examined for abnormalities and the organs removed, examined, and, where required, placed

in fixative. Special emphasis was placed on or gans of the reproductive system. Implantation

sites (scars) along the uterus were counted and recorded. All de signated tissues were fixed in

neutral buffered formalin, except for the testes and epididymides, which were placed in

Bouin’s fixative.

4.4.2. Organ Weights

Body weights and protocol-desi gnated organ weights were reco rded for all surviving adult

animals at the scheduled necropsy and appropr iate organ weight ratios were calculated

(relative to body weight). Paired organs were weighed together. The right and left testis

were weighed separately, and a combined we ight is summarized in the report. Page 19 of 358

# 1271-001 4.4.3. Microscopic

Microscopic examination of fixed hematoxylin and eosin-stained paraffin sections was

performed on protocol-designated sections of tissues. The slides for the control and

2400 mg/kg/day group were examined by a board -certified veterinary pathologist. A

four-step grading system was utilized to define gradable lesions for comparison between dose

groups. A glossary of macroscopic and microsco pic terms used in this study is included in

this report. A full list of organs and tissues collected, weighed, and examined for this study

is presented in Appendix I.

4.5. Statistics

The table below defines the set of comparisons us ed in the statistical analyses described in

this section.

Table of Statistical Comparisons

Control Group Treatment Groups

1 2, 3, 4

Data for each sex within a set were analyzed separately.

The raw data were tabulated within each time interval, and the mean and standard deviation

were calculated for each endpoint by sex and group. For each endpoint, treatment groups

were compared to the control group using the analysis outlined belo w and on the following

page.

Endpoint Analysis

Parental In-life Data

Prematin g Bod y Wei g hts Group Pai r -wise Comparisons

Premating Food Consumption Group Pair-wise Comparisons

Gestation Body Weights Group Pair-wise Comparisons

Gestation Body Weight Change s Group Pair-wise Comparisons

Gestation Food Consumpti on Group Pair-wise Comparisons

Lactation Body Weights Group Pair-wise Comparisons

Lactation Body Weight Cha nges Group Pair-wise Comparisons

Lactation Food Consumption Group Pair-wise Comparisons

Postmating Body Weights - Ma les Group Pair-wise Comparisons

Postmating Food Consumption - Males Group Pair-wise Comparisons

Fertilit y Indices

Estrous C y cle (mean c y cle time) Group Pai r -wise Comparisons

Estrous Cycle (# cycles/per iod) Group Pair-wise Comparisons

Gestation Length Group Pair-wise Comparisons

Copulatory Interval Gr oup Pair-wise Comparisons Page 20 of 358

# 1271-001 End p oint Anal y sis

Male Fertilit y Index Fisher’s Exact Test

Female Fertility Index Fisher’s Exact Test

Female Mating Index Fisher’s Exact Test

Male Mating Index Fisher’s Exact Test

Female Fecundity Index Fisher’s Exact Test

Male Fecundity Index Fisher’s Exact Test

Gestation Index Fisher’s Exact Test

Patholo gy

Male Reproduction Organ Wei ghts Group Pair-wise Comparisons

Female Reproduction Organ Wei ghts Group Pair-wise Comparisons

Litter

Number of Implantation scars/dam Group Pai r -wise Comparisons

Litter Size Group Pair-wise Comparisons

Viable Pups Group Pair-wise Comparisons

Pup Sex Ratio (% viable males/litt er) Arcsin-Square Root Transformation

Stillborn Pups Group Pair-wise Comparisons

Stillborn Index Arcsin -Square Root Transformation

% Postimplantation Loss Arcs in-Square Root Transformation

Pup Weights Covariate Analysis

Pup Survival (Days 0-4) Ar csin-Square Root Transformation

4.5.1. Group Pair-Wise Comparisons (Levene’s/ANOVA-Dunnett’s/Welch’s)

If sample sizes for all groups were 3 or greater, Levene’s test 4 

# was used to assess

homogeneity of group variances for each specified endpoint and for all collection intervals. If

Levene’s test was not significant (p> 0.01), a pooled estimate of the variance (Mean Square

Error or MSE) was computed from a one-way analysis of variance (ANOVA) and utilized by

a Dunnett’s 5 

# comparison of each treatment group with the control group. If Levene’s test

was significant (p<0.01), comparisons with th e control group were made using Welch’s

t-test 6 

# with a Bonferroni correction.

In the case that sample size was less than 3 for at least one treatm ent group, Levene’s method

could not be implemented. Groups with sample sizes less than 3 we re excluded from the

analysis and control-treatment pair-wise comparisons that satisfied the sample size

assumption (n> 3) were conducted using Welch’s t-te st with a Bonferroni correction.

If there were only 2 groups involved, the above methodology applied and the Dunnett’s test

reduced to a Student’s t-test 7



# .

4 

# Milliken GA, Johnson DE. Analysis of messy data. London: Chapman and Hall: 1992.

5 

# Dunnett CW. A multiple comparison procedure for comparing several treatments with a control. J Am Stat

Assoc 1955;50:1096-1121.

6 

# Welch BL. The significance of di fference between two means when the population variances are unequal.

Biometrika 1937;29:350-362. 

# Page 21 of 358

# 1271-001 Results of all pair-wise compar isons were reported at the 0.05 and 0.01 significance levels.

All endpoints were analyzed using two-tailed tests.

4.5.2. Arcsin-Square-Root Transformation Follo wed by Group Pair-wise Comparisons

Data comprised of percent values were tran sformed using the arcsin of the square root 7 

# . The

analysis described in the Group Pair-wise Co mparison section was used to analyze the

transformed percentage values.

4.5.3. Fisher’s Exact Test

An overall test for association between re sponse and treatment was conducted using a

Fisher’s exact 8 

# test. If this overall test was significant (p < 0.05) and there were more than

two groups, then a follow up analysis was conducted where each treatment group was

compared to the control group using a Fisher’s exact test. Results we re reported at the 0.05

and 0.01 significance levels. All endpoints were analyzed using two-tailed tests.

Overall testing and follow up pair-wise tes ting were conducted for the control and all

treatment groups that had sufficient sample size (n ≥ 3). If all groups had an insufficient

sample size (n<3), or if just the control group had a sample size of less than 3, no testing was

performed.

4.5.4. Covariate Analysis

For groups with sample sizes greater than or equal to 3, an analysis of covariance 9 

# was

conducted to compare treatment groups for each endpoint (mean of pup body weights per

dam) and time period. The covariate was litter si ze. If the control gr oup did not satisfy the

sample size assumption (n<3), no further anal ysis was performed. Each treatment group

satisfying the sample size assumption was compar ed to control using Du nnett’s test under the

analysis of covariance model.

The LSMEANS (Least Square Means), which are the means adjusted for values of the

covariate, are displayed on the summary table.

Results of pair-wise comparisons were repor ted at the 0.05 and 0.01 si gnificance levels.

Endpoints were analyzed using two-tailed tests.

7 

# Steel RGD, Torrie JH. Principles and Procedures of Statistics. A biometrical approach. New York: McGrawHill; 1980.

8 

# Zar JH. Biostatistical Analysis. 4 th 

# ed. New Jersey: Prentice Hall; 1999.

9 

# Snedecor, GW, Cochran, WG. Statistical Methods, Ei ghth Edition. Ames (IA): Iowa State Press; 1989. 

# Page 22 of 358

# 1271-001 4.6. Computer Systems

The computer systems used during the conduct of this study are presented in the following

table.

MPI Research Computer Systems

Provantis TM 

# v6.5: Client-server, Oracle-based system primarily used for

toxicology studies.

Dispense v5.0: Automates the test material control processes.

Niagara Framework ® 

# Software System

v2.3: Environmental monitoring, alarming, and reporting

application.

Millennium 32 

# v4.0: Millennium 32 

# HPLC System (instrument) and

Millennium 32 

# Chromatography Manager Version

(software) used to quantitatively determine the amount of

test article in prepared test article batches.

MPI Archiving System (MArcS) v1.1: In-house de veloped application for automated storage

and retrieval information for archiveable materials (e.g.,

lab books, study data, wet tissues, slides, etc.).

MPI Reporting System (MPIRS) v 6.5

and v7.0: In-house developed reporting system running SAS ®



# programs primarily reporting on Provantis data.

Master Schedule v2.2: Maintains the master schedule for the company.

SAS ® 

# v8.2: The SAS ® 

# System is an integrated system of software

products that enables a user to perform data entry,

retrieval, data management, reporting, graphics,

statistical analysis, and a pplications development.

Microsoft ® 

# Office 2003 Professional: Bundle of integrated produc tivity tools including word

and data processing and co mmunications software.

Contains the utilities Microsoft ® 

# Access, Excel, InfoPath,

Outlook, PowerPoint, Publisher, and Word.

Microsoft ® 

# Office 2000 Professional: Microsoft ® 

# Excel 2000 used in conjunction with

Millennium 32 

# software.

Additional information is available in the MPI Research, Inc., company document titled

“Computer Systems Information.”

4.7. Data and Specimen Retention

All raw data, documentation, records, protocol , reserve samples, specimens, and the final

report generated as a result of this study will be retained at MPI Research, Inc., or an

approved archive facility contracted by MPI Rese arch, Inc., for a period of 1 year following

completion of the study (final repor t issue date). Retention of ma terials after the time stated

above will be subject to futu re contractual agreements. Page 23 of 358

# 1271-001 5. RESULTS AND DISCUSSION

5.1. Analysis of Dosing Formulations

The individual results of the dosing formula tion analysis for hom ogeneity, stability, and

concentration are presented in Appendix B and are summarized below.

Preparation of the dosing suspen sions was problematic. Due to the viscosity of the vehicle

(1% Guar Gum), the formulations were unable to be stirred adequatel y, and therefore it was

difficult to obtain uniform suspensions. Periodi c concentration analysis of the test article

suspensions determined that the formulations for the most part were neither homogenous nor

were the intended dose levels achieved.

5.1.1. Homogeneity

Homogeneity analysis of the test article su spensions was conducted at the low and high

concentrations prior to study init iation (first test ba tch) and mean recoveries were low at 55.5

and 76.3% of nominal for the 150 and 2400 mg/kg/ day levels, respectively. Homogeneity

samples were again collected and analyzed fr om the second test batch preparation. Mean

recovery for the 150 mg/kg/day level was ag ain low at 50.4%, however, the 2400 mg/kg/day

level was within specifications ( ± 15%) at 87.5%. Due to a change in preparation

instructions (material prepared on a we ight/weight basis beginning on July 11, 2006)

additional homogeneity samples were collected and analyzed during Week 3. At this time

mean recovery for the 150 mg/kg/day level was 83.1% and recovery for the 2400 mg/kg/day

level was 91.4%.

The data are summarized below.

Homogeneity (Test Batch 1)

Dose Level

(mg/kg/day) Nominal

Concentration

(mg/mL) Mean Found

Concentration

(mg/mL) Average

Measured

Dose Level

(mg/kg/day) Mean %

Recovery a 

# RSD

(%)

150 7.5 4.1661 83.3220 55.5 26.5

2400 120 91.5363 1830.7260 76.3 7.2

a 

# Mean % recovery was calculated from the nominal concentration.

RSD - Relative Standard Deviation Page 24 of 358

# 1271-001 Homogeneity (Test Batch 2)

Dose Level

(mg/kg/day) Nominal

Concentration

(mg/mL) Mean Found

Concentration

(mg/mL) Average

Measured

Dose Level

(mg/kg/day) Mean %

Recovery a 

# RSD

(%)

150 7.5 3.7765 75.5300 50.4 20.0

2400 120 104.9651 2099.3020 87.5 8.5

a 

# Mean % recovery was calculated from the nominal concentration.

RSD - Relative Standard Deviation

Homogeneity (Week 3)

Dose Level

(mg/kg/day) Nominal

Concentration

(mg/mL) Mean Found

Concentration

(mg/mL) Average

Measured

Dose Level

(mg/kg/day) Mean %

Recovery a 

# RSD

(%)

150 7.5 6.2297 124.5940 83.1 12.0

2400 120 109.6368 2192.7360 91.4 13.7

a 

# Mean % recovery was calculated from the nominal concentration.

RSD - Relative Standard Deviation

5.1.2. Stability

Stability analyses were conducted for the test batch and for Week 1 and 3 mixes along with

homogeneity evaluations. The test formulati ons at the low (150 mg /kg/day) and high (2400

mg/kg/day) dose levels were considered stable over the period of use.

5.1.3. Concentration

Samples were collected from the Week 1, 3, and 8 mixes and analyzed for test article

concentration. Due to the viscosity of the test ar ticle formulations and the inability to stir the

mixes adequately to keep the Anthraquinone in suspension with the vehicle (1% Guar Gum),

recoveries at all levels were relatively low throughout the st udy period. Recoveries ranged

from 66.0-73.0% (Week 1), 59.5-84.7% (Week 3), and 25.1-99.5% (Week 8) in the treated

groups. Based on these recoveries the aver age measured dose levels were 109, 396, and

1661 mg/kg/day for Week 1, 116, 357, and 2034 mg/kg/day for Week 3, and 38, 463, and

2388 mg/kg/day for Week 8, respectively, fo r the 150, 600, and 2400 mg/kg/day dose levels.

These data are summarized on the following page . No test article was seen in the control

samples. Page 25 of 358

# 1271-001 Concentration (Week 1)

Dose Level

(mg/kg/day) Nominal

Concentration

(mg/mL) Mean Found

Concentration

(mg/mL) Average Measured

Dose Level

(mg/kg/day) % Recovery a



# 0 0.0 NA NA NA

150 7.50 5.4747 109.4940 73.0

600 30.00 19.8070 396.1400 66.0

2400 120.00 83.0470 1660.9400 69.2

a 

# Mean % recovery was calculated from the nominal concentration.

NA – Not Applicable

Concentration (Week 3)

Dose Level

(mg/kg/day) Nominal

Concentration

(mg/mL) Mean Found

Concentration

(mg/mL) Average Measured

Dose Level

(mg/kg/day) % Recovery a



# 0 0.0 NA NA NA

150 7.50 5.8127 116.2540 77.5

600 30.00 17.8468 356.9360 59.5

2400 120.00 101.6758 2033.5160 84.7

a 

# Mean % recovery was calculated from the nominal concentration.

NA – Not Applicable

Concentration (Week 8)

Dose Level

(mg/kg/day) Nominal

Concentration

(mg/mL) Mean Found

Concentration

(mg/mL) Average Measured

Dose Level

(mg/kg/day) % Recovery a



# 0 0.0 NA NA NA

150 7.50 1.8801 37.6020 25.1

600 30.00 23.1492 462.9840 77.2

2400 120.00 119.4088 2388.1760 99.5

a 

# Mean % recovery was calculated from the nominal concentration.

NA – Not Applicable Page 26 of 358

# 1271-001 5.2. In-life Examinations

5.2.1. Mortality

A record of animal fate and disposition is presented in Appendix C.

Mortality was largely restrict ed to females at the higher dose levels. In the 600 and

2400 mg/kg/day groups eight and se ven females, respectively, died or were euthanized

in extremis. These deaths were considered test article -related. At necrops y, black or red foci

were observed in the glandular stomach of se veral of these females (three females at

600 mg/kg/day and four females at 2400 mg/ kg/day). In addition, four females at

2400 mg/kg/day were observed at gross examinati on or microscopically w ith erosion/ulcer of

the stomach. All females in the control a nd 150 mg/kg/day group survived to terminal

euthanasia.

One male in the 150 mg/kg/day group was found dead during the study, but necropsy

findings were inconclusive as to the cause of death and in the abse nce of mortality among

males at the higher dose levels, this death was not considered te st article-related.

5.2.2. Clinical Findings

Clinical findings are summarized in Tables 1 (males), 2 (females – premating/mating), and

3 (females - gestation) and presen ted individually in Appendix D.

Test article-related clinical fi ndings of discolored urine and d ecreased activity were noted in

all treatment groups during the study period. Al l males and females in the treated groups

were observed at one or more da ily intervals with discolored ur ine (brown, orange, or red).

In addition, decreased activity was observed in four and five females in the 600 and

2400 mg/kg/day groups, respectively, and one male at 150 mg/kg/day. There was an

occasional occurrence of discolored (orange) urine noted among the controls but decreased

activity was not seen. A few additional diss imilar observations were noted at clinical

examination of the treated animals but these we re seen either at a low frequency or with

similar frequency as controls and were considered unrelated to treatment with

Anthraquinone.

5.2.3. Body Weights and Body Weight Changes

Body weight data are summarized in Tables 5 (males), 6 (fem ales – premating/mating), 7

(females – gestation), and 8 (f emales – lactation). Body wei ght change data are summarized

in Tables 9 (females – gestation) and 10 (femal es – lactation). Body weight and body weight

change data are presente d individually in Appendices E and F, respectively.

Mean body weights of treated males were lo wer than controls th roughout the premating,

mating, and postmating period and at 600 and 2400 mg/kg/day many of these differences

were statistically significant. Mean body weights of treated fe males were also lower than

control throughout the premating, mating, gest ation, and lactation period and in most

instances these differences too were statistically significant. Gestati on body weight gain was

significantly lower than contro l over most intervals (GD 0-7, 7-14, 14-20, and 0-20) in the

600 and 2400 mg/kg/day groups, and although not st atistically significan t, body weight gain Page 27 of 358

# 1271-001 was also lower than controls over GD 14-20 and 0-20 in the 150 mg/kg/day group. During

the lactation period (LD 0-4), body weight gain for the few females that retained pups in the

600 and 2400 mg/kg/day groups was lower than controls, although these differences were not

statistically significant. B ody weight gain in the 150 mg/ kg/day group during lactation was

comparable to controls.

5.2.4. Food Consumption

Food consumption data are summarized in Tabl es 11 (males), 12 (females – premating), 13

(females – gestation), and 14 (fem ales – lactation) and presente d individually in Appendix G.

A treatment-related effect on food consumption was evident in all treat ed males and females

in comparison to controls, although these re ductions were not dose-dependent. Food

consumption for all treated males was signifi cantly lower than controls during Week 1-2

(premating period) and was signi ficantly reduced in males at the 600 mg/kg/day level during

Week 5-7 (postmating period) and in the 150 mg/kg/day group during Week 6-7. Premating,

gestation, and lactation food consumption was lowe r than control in all treated female groups

and in most instances these differences were statistically significant.

5.3. Postmortem Study Evaluations

The pathology interpretation of the results is presente d below and in Appendix H.

5.3.1. P Macroscopic

Macroscopic observations are summarized in Tabl es 15 (males) and 16 (females). Individual

macroscopic observations are presented in Appendix K.

Test article-related macroscopic changes we re noted in female rats that received

Anthraquinone (CAS # 84-65-1), but not in males. Black or red foci were observed in the

glandular stomach of females at 600 and 2400 mg /kg/day that died on study (DOS) and an

erosion/ulcer was noted in the pyloric stomach of one DOS female at 2400 mg/kg/day. Black

or red foci and the erosion/ulcer corresponded microscopically with erosion/ulcer of the

glandular stomach.

Other macroscopic changes noted were typical of those commonly seen in rats of the same

strain and age or did not demonstrate a dose-re lated response, and were considered to be

incidental.

5.3.2. P Organ Weights

Organ weight values are summarized in Tables 17 (males) and 18 (females). Individual

organ weight values are presented in Appendix J.

No test article-related organ weight cha nges were noted in rats of either sex.

In males, the relative (to body weight) seminal vesicle weight at 2400 mg/kg/day and relative

(to body weight) left and right testes weights at 600 and 2400 mg/kg/day were statistically Page 28 of 358

# 1271-001 significantly increased compared to the contro ls, but the relative increases were caused by

decreased body weight and were no t primary organ weight changes.

In females, statistically significant decreases were noted in the mean weight of the ovaries at

2400 mg/kg/day and the mean weight of the uterus at 600 and 2400 mg/kg/day. Decreased

weight of the ovaries and uterus was consider ed to be secondary to significantly decreased

body weights in these groups and not primary organ weight changes. 10



# 5.3.3. P Microscopic

Microscopic observations are summ arized in Tables 19 (males) and 20 (females). Individual

microscopic observations are presented in Appendix K.

Test article-related microscopic changes were noted in the liver and glandular stomach of

female rats, but not in males.

Minimal focal necrosis and mode rate periportal vacuol ation of hepatocytes were observed in

one female at 2400 mg/kg/day. Edema (2 of 7) a nd erosion/ulcer (3 of 7) were present in the

glandular stomach of DOS females at 2400 mg/kg/day.

Other microscopic changes noted were typical of those commonly seen in rats of the same

strain and age or had a similar incidence in tr eated and control rats, a nd were considered to

be incidental.

5.4. Reproductive Performance

5.4.1. P Estrous Cyclicity

Estrous cyclicity data are summarized in Table 21, and presented individually in Appendix L.

Mean estrous cycle length (number of days per cycle) was increased in the treated females

(4.7, 5.6, and 4.8 days in the 150, 600, and 2400 mg/kg/day groups, respectively) in

comparison to controls (4.4 days), but was onl y statistically signifi cant at 600 mg/kg/day.

Therefore, with the increase in cycle length, the number of cycl es per animal over the 2-week

evaluation period was significantly decreased in all treated groups ( 2.5, 2.2, and 2.7 cycles

per animal in the 150, 600, and 2400 mg/kg/da y groups, respectively) in comparison to

controls (3.6 cycles per animal).

5.4.2. P Reproductive and Fertility

Reproductive and fertility indices are summarized in Table 22. Individual mating records are

presented in Appendix M

The mating, fertility, and fecundity indices ( 83.3-100%) for both treated males and females

were unaffected by treatment with Anthraqu inone and were similar to controls

10 

# Levin S, Semler D, and Ruben Z. Effects of two weeks of feed restriction on some common toxicologic

parameters in Sprague-Dawley rats. Toxicologic Pathology, 21(1): 1-14, 1993. 

# Page 29 of 358

# 1271-001 (91.7-100.0%). Likewise, the copulatory inte rval (days to mating) was unaffected by

treatment and ranged from 2.4-3.1 days in the treated groups in comparison to 3.0 in the

controls.

5.5. P Delivery and F 1 

# Litter Data

Natural delivery and litter data are summarized in Table 23 and presented individually in

Appendix N.

There were 11, 10, 12, and 11 females pregnant in the control, 150, 600, and 2400 mg/kg/day

groups, respectively. All pregnant animal s in the control an d 150 mg/kg/day groups

delivered and retained litters through LD 4. In the 600 mg/kg/day group 10 females

delivered, but only two females retained litter s through LD 4. Two females died pregnant,

two females died during delivery, four female s died during the lact ation period, and two

females lost their entire litter prior to LD 4. In the 2400 mg/kg/day group four females

delivered, but only one female retained a litte r through LD 4. Four females died pregnant,

one pregnant female was euthanized in extremis , one female had all resorbed fetuses, and

three females lost their en tire litter prior to LD 4.

The number of pups delivered (live + dead) was significantly decreased in the 600 and

2400 mg/kg/day groups. Likewise, the number of liveborn pups was decreased with an

increase in stillborn pups in these same groups. The still born index (mean % pups/litter),

although not significant, was 19.97% and 26.04% in the 600 and 2400 mg/kg/day groups,

respectively, compared to no stillborn pups in the control group. M ean number of total

implantation scars/litter was statistically significantly lower at 600 and 2400 mg/kg/day

(13.0 and 12.5, respectively) in comparison to controls (15.5 scars). Additionally, mean

post-implantation loss at 600 and 2400 mg /kg/day was higher at 20.48% and 31.09%

(statistically significant), respectively, compared to controls (6.07%). These effects (i.e.,

decrease in mean number of to tal implantation scars, fewer nu mber of pups delivered (live

plus stillborn), and increase in post-impla ntation loss) at 600 a nd 2400 mg/kg/day were

considered test article-related. The number of pups born, liveborn, and stillborn in the

150 mg/kg/day group was similar to contro ls and unaffected by treatment with

Anthraquinone.

5.6. F 1 

# Pup Evaluations

5.6.1. Clinical Observations

F 1 

# pup clinical findings are summarized in Table 24 and presented individually in

Appendix O.

A few dissimilar clinical observations were noted among pups in the 150 mg/kg/day group,

but these were seen at a low frequency and considered unrelat ed to treatment. Several pups

were noted with skin cold to touch in th e 600 mg/kg/day group. Th ese observations were

noted on LD 0 and all of these pups died prior to LD 4. Very few pups survived to LD 4 in

the 2400 mg/kg/day group, but no clinical ob servations were noted for these pups. Page 30 of 358

# 1271-001 5.6.2. Body Weights

F 1 

# pup body weight values are summarized in Table 25 and presented individually in

Appendix P.

Mean pup body weights (by sex and sexes co mbined) in the 150 mg/kg/day group were

similar to controls on LD 0, but by LD 4 were approximately 8-10% lower than controls. In

the 600 mg/kg/day group these weights were si gnificantly lower (22-25%) in comparison to

controls on LD 0. Only two lit ters remained on LD 4 for this group, and although statistical

analysis could not be conducted due to the sma ll litter size, weights were 51-58% lower than

controls. Due to the small number of litters in the 2400 mg/kg/day group, statistical analysis

was not conducted for pup body weights, but weights on LD 4 were 10-11% lower than

controls.

5.6.3. Pup Survival

Pup survival indices during lactation are summarized in Table 23 and derived from

individual litter da ta in Appendix N.

Pup survival (LD 0-4) in the 150 mg/kg/day group was 89% and was comparable to the

controls at 97%. Pup survival, although not dose-dependent, for the 600 and 2400 mg/kg/day

groups was significantly lower (16.5 and 33.3%, respectively) in comparison to controls.

5.6.4. Pup Sex Ratio

F 1 

# pup sex ratios are summarized in Table 23 and the distribution of sexes in individual litters

during lactation is presented in Appendix P.

Pup sex ratio (% males per litter) in the treated groups was consid ered comparable to controls

and unaffected by treatment.

5.6.5. Pup Macroscopic

Pup macroscopic observations are summarized in Table 26 an d presented individually in

Appendix Q.

No macroscopic observations were evident in an y pups (stillborn, died on study, or LD 4) at

necropsy. Page 31 of 358

# 1271-001 6. CONCLUSION

In this reproduction/developmental toxici ty screening study in rats administered

Anthraquinone orally, neither a paternal, mate rnal, nor a developmental No-Observed-EffectLevel (NOEL) was achieved at the dose levels ev aluated. Both male and female rats at all

treatment levels (150, 600, and 2400 mg/kg/day) were observed with discolored urine, and

decreased body weights and food consumption. C onsiderable mortality was seen in females

at the 600 and 2400 mg/kg/day levels and ulce r/erosion of the stomach was evident at

necropsy in the 2400 mg/kg/day group. Mating, fertility, and fecundity indices (male and

female) in the treatment groups were comparable to controls, however, several females in the

600 and 2400 mg/kg/day groups died during the gestation and lact ation period. Other effects

seen at parturition at these dose levels were a decrease in pups born, fewer liveborn,

increased in stillborn pups, a nd increased post-implan tation loss. Pup su rvival was poor in

the 600 and 2400 mg/kg/day groups with only two a nd one litter, respectively, surviving to

LD 4. In addition, pup body weights were decreased at all treatment levels. Page 32 of 358

# 1271-001 REPRODUCTION GLOSSARY

Abnormality - A morphologic or functional

deviation from normal limits (anomaly).

Activity - Action or process of exerting energy.

Autolysis - Decomposition occurring

spontaneously after death.

Conception - The onset of pregnancy; formation

of a visible zygote.

Delivery - Expulsion of offspring at birth.

Development - Gradual growth or expansion,

especially from a lower to a higher stage of

complexity.

- Arrested Development -

cessation of the developmental process at

some stage prior to its normal completion.

- Prenatal Development -

That which occurs before birth.

- Postnatal Development -

That which occurs after birth.

Developmental Toxicity - Toxicity incurred by

the conceptus during development. There are four

manifestations: (1) growth retardation, (2) death,

(3) terata or malformation, and (4) functional

deficit. The term supercedes older terms

“embryotoxicity” (death, malformation) and

“fetotoxicity” (death, growth retardation), thereby

eliminating temporal considerations.

Distal - Distant from the trunk.

Embryo - The early or developing stage of an

organism, especially the developing product of

fertilization of an egg. Its development is termed

embryogenesis. The embryonic period is as

follows:

Rat 6-13 days postconception

Fecundity Index or Pregnancy Index - Males

No. males impregnating a female x 100

No. males with evidence of mating

Fecundity Index or Pregnancy Index - Females

No. females pregnant x 100 No. females with evidence of mating

Females Delivering Litters (%)

No. females delivering litters x 100

No. females pregnant

Females with All Stillborn (%)

No. litters with all stillborn pups x 100

No. litters with at least 1 stillborn pup

Females with Stillborn Pups (%)

No. litters with at least 1 stillborn pup x 100

No. females delivering litters

Fertility Index - Males

No. males impregnating a female x 100

No. males paired

Fertility Index - Females

No. females pregnant x 100

No. females paired

Fetus - The unborn offspring; the developing

young following embryogenesis. The fetal period

is as follows:

Rat 15-22 days postconception

Gestation - The period of intrauterine

development, conception to birth.

Gestation Day 0 - Day when positive evidence of

mating has been ascertained.

Gestation Duration - The length of gestation;

term of pregnancy. The duration is as follows for

the common species:

Rat 21 days

Gestation Index

– No. females delivering at least 1 live pup x 100

No. females pregnant

GLPs - Good Laboratory Practices. Legislation

first enacted in June, 1979 to assure the quality

and integrity of safety data filed pursuant to

products regulated by the U.S. FDA. The Final 

# Page 33 of 358

# 1271-001 Rule became effective October 5, 1987 [ Fed. Reg.,

52 (172: 33768, 1987)]. Similar GLPs have since

been adopted by the U.S. EPA (effective October

16, 1989 for pesticides, and September 18, 1989

for TSCA-regulated chemicals), Japan Koseisho

(April 1, 1983) and Japan MAFF (October 1,

1984).

Gravid - An animal with uterine evidence of

implantation which delivers a conceptus or

contains developing young (pregnant).

Impaired - Diminished or reduced.

Implantation - Attachment of the blastocyst to the

epithelial lining of the uterus, its penetration

through the epithelium, and its embedding in the

compact layer of the endometrium (nidation).

It occurs after fertilization as follows:

Rat 5.5 - 6 days

Implantation sites not discernible by placental

remains may be visualized by staining of the

uterus with ammonium sulfide (Kopf method,

Naunyn Schmiedebergs Arch. Pharmacol., 247:

121-135, 1964).

The types of implants as defined by SOP are as

follows: a) viable fetus - responds to touch, b)

nonviable fetus - does not respond to touch, no

signs of autolysis, c) late resorption - recognizable

fetal form, but undergoing autolysis, d) early

resorption - implantation site, tissue has no

recognizable fetal characteristics.

Lactation Index – Day 21

No. pups surviving 21 days (weaning) x 100

No. live pups at 4 days (postculling)

Lesion - Any pathological or traumatic

discontinuity of tissue or loss of function of a part;

a circumscribed area of pathologically altered

tissue.

Litter Size - In polytocous animals, pregnancy

consists of multiple offspring in the litter.

Live Birth Index –

No. live pups at birth x 100

No. pups born Malformation - Defective or abnormal function;

anatomic or morphologic abnormality; deformity

(dysmorphosis, cacomorphosis). Defined recently

by a regulatory agency as “a permanent deviation

which generally is incompatible with or severely

detrimental to normal postnatal survival or

development” [EPA, Fed.Reg., 51 (185): 34028,

1986].

For practical purposes we use the following

definition in this laboratory: Malformations are

those structural anomalies that alter general body

conformity, disrupt or interfere with body

function, or are generally thought to be

incompatible with life. Specific examples of

processes that result in maldevelopment include

marked or severe misshapening, asymmetry or

irregularity of structure brought about by fusion,

splitting, disarticulation, malalignment, hiatus,

enlargement, lengthening, thickening, thinning, or

branching. Absence (agenesia) of parts or whole

structures is also considered a malformative

process.

Malformations occur when normal organogenesis

is interrupted.

Mating Index - Males

No. males with evidence of mating x 100

No. males paired

Mating Index - Females

No. females with evidence of mating x 100

No. females paired

Moribund - In a state of dying.

NOEL - No observable effect level.

NOAEL - No observable adverse effect level.

Nonviable - A nonliving conceptus; includes early

and late resorptions and dead fetuses.

Postimplantation Loss –

No. implantation scars - No. pups delivered x 100

No. implantation scars 

# Page 34 of 358

# 1271-001 Reproductive Effects - Those findings associated

with reproductive parame ters in generation

studies. They are usually classed as primary

(effects related to sex organs or reproductive

capacity of adults), or secondar y (changes that

may be dependent upon or re lated to other signs of

toxicity).

A tabulation of primary reproductive effects might

include (Christian, J. Am.Coll. Toxicol., 5: 161,

1986):

- estrous

- mating performance

- fertility

- fecundity

- duration of gestation

- delivery complications

- pathology of reproductive

organs

- litter size

- litter viability at birth

- litter survival

- sex ratios of litters

- growth & functional

development of litter, birth

to weaning

SOPs - Standard Operating Procedures.

Spontaneous Malformations - The normal

background incidence of maldevelopment

unrelated to known causes. The rate of

spontaneous malformation for common species is

as follows (Schardein, Chemically Induced Birth

Defects, Dekker 1993).

Rat 0.02 - 2%

Stillborn Index –

No. stillborn pups x 100

No. pups born

Teratogen - An agent or factor that causes the

production of physical defects in the developing

embryo. The production of defects is termed

teratogenesis

.

Ventral - Pertaining to the belly.

Viable - Capable of living. Viability Index – Day 4

No. pups surviving 4 days (preculling) x 100

No. live pups at birth 

# Page 35 of 358

# PATHOLOGY GLOSSARY

1271-001 Grading System:

Minimal - The amount of change present

barely exceeds that which is considered to

be within normal limits (10% or less of the

tissue is involved).

Mild - In general, the lesion is easily

identified but of limited severity. The

lesion probably does not produce any

functional impairment (11 - 20% of the

tissue is involved).

Moderate - The lesion is prominent but

there is significant potential for increased

severity. Limited tissue or organ

dysfunction is possible (21 - 40% of the

tissue is involved).

Severe – The degree of change is either as

complete as considered possible or great

enough in intensity or extent to expect

significant tissue or organ dysfunction

(41 - 100% of the tissue is involved).

Abrasion - A loss of superficial layers of

the skin or a mucous membrane secondary

to friction.

Adhesion - The binding together of

adjacent organs or tissues by cells or cell

products. The adhesions may be formed

from the proliferation of fibroblasts with

collagen production, from fibrin exuded in

response to an irritant, or from the

presence of fibrin on the serosal surface of

the adhered organs or tissues.

Body fat depleted - The loss of visible fat

from the body, especially in subcutaneous

tissue, retroperitoneal space, abdominal

cavity and/or mediastinum. Discoloration - Any color change that

differs from the normal.

Discoloration, pink - A change in color

from normal or from surrounding tissue to

a pinkish hue.

Discoloration, red - A change in color

from normal or from surrounding tissue to

a reddish hue.

Edema - Excessive accumulation of tissue

fluid is present in intercellular tissue

spaces.

Erosion/ulcer – Necrosis and loss of

architecture of the epithelium constitutes

an erosion. If the basement membrane is

disrupted the lesion is an ulcer.

Focus/foci, black - One or more discrete

areas exhibiting a black coloration, which

is/are different from the color of the

surrounding tissue and which is/are flush

with or depressed relative to the

surrounding tissue.

Focus/foci, red - One or more discrete

areas exhibiting a reddish coloration,

which is/are different from the color of the

surrounding tissue and which is/are flush

with or depressed relative to the

surrounding tissue.

Infiltration, lymphocytic - The presence

of increased numbers of lymphocytes

within a tissue.

Infiltration, mononuclear cell - The

presence of increased numbers of cells

with nonsegmented nuclei, particularly

lymphocytes, plasma cells and/or

macrophages, within an organ or tissue. Page 36 of 358

# PATHOLOGY GLOSSARY

1271-001 Lobulation, accentuated - In the liver, an

increase in the prominence of the

individual hepatic lobules as viewed from

the serosal and/or cut surface of the organ.

Necrosis, focal – There is a focal area of

dead tissue present.

Pale - Whitish or pallid; lighter in color

than normal.

Small - Comparatively little in size or

dimension; not as large as normally

expected. Vacuolation, periportal – The presence of

cytoplasmic vacuoles primarily in

hepatocytes surrounding the portal triads

of hepatic lobules.

Within normal limits - Tissue considered

to be normal, under the conditions of the

study and considering the age, sex, and

strain of the animal concerned. Alterations

may be present which, under other

circumstances, would be considered

deviations from normal. Page 37 of 358

# 1271-001 Table 1

Summary of P Clinic al Findings - MALE Page 38 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 1 Summary of P Clinical Findings+- MALE0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedNumber of Animals Alive at Start of Interval12 12 12 12Behavior/ActivityActivity decreased 0/0 3/1 0/0 0/0Salivation 0/0 0/0 1/1 0/0ExcretionUrine discolored, Brown 0/0 0/0 5/2 0/0Urine discolored, Orange 2/1 40/12 51/12 59/12Urine discolored, Red0/0 66/12 213/12 207/12External AppearanceCarriage low 0/0 2/1 0/0 0/0Malocclusion 0/0 0/0 8/1 0/0Material around eyes, Black, Eye/left 0/0 0/0 4/1 0/0Material around eyes, Black,Eye/right 0/0 0/0 4/1 0/0Material around eyes, Red, Eye/left 0/0 0/0 4/1 0/0Material around eyes, Red,Eye/right 0/0 0/0 4/1 0/0Material around nose, Black 0/0 0/0 0/0 1/1Material around nose,Red 0/0 2/1 0/0 0/0Posture hunched0/0 9/1 0/0 0/0Swelling, Anogenital region 0/0 0/0 0/0 12/1Pelage/SkinAbrasion(s), Cervical region 0/0 0/0 21/1 0/0Abrasion(s), Lip 0/0 0/0 0/0 1/1Hair discolored, Brown, Anogenital region 0/0 0/0 0/0 2/1Hair discolored, Brown, Forelimb/left 0/0 15/1 0/0 0/0Hair discolored, Brown, Forelimb/right 0/0 15/1 0/0 0/0Hair discolored, Brown, Ventral surface 0/0 0/0 0/0 18/1Hair discolored, Yellow, Anogenital region 1/1 0/0 0/0 0/0 

# Page 39 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 1 Summary of P Clinical Findings+- MALE0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedPelage/SkinHair discolored, Yellow, Ventral surface 0/0 0/0 0/0 1/1Hair sparse, Cervical region 0/0 0/0 11/1 0/0Hair sparse, Face 0/0 11/1 0/0 0/0Hair sparse, Forefoot/left 25/1 11/1 0/0 11/1Hair sparse, Forefoot/right 25/1 11/1 0/0 11/1Hair sparse, Forelimb/left 0/0 36/2 0/0 0/0Hair sparse, Forelimb/right 0/0 36/2 21/2 0/0Hair sparse, Ventral surface 0/0 9/1 0/0 0/0Piloerection 0/0 12/1 0/0 0/0Scabbed area, Thoracic region 0/0 0/0 7/1 0/0RespirationBreathing difficult0/0 3/1 0/0 0/0Rales 0/0 0/0 0/0 1/1 

# Page 40 of 358

# 1271-001 Table 2

Summary of P Premating/Mati ng Clinical Findings – FEMALE Page 41 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 2 Summary of P Premating/MatingClinical Findings+- FEMALE0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedNumber of Animals Alive at Start of Interval12 12 12 12Behavior/ActivityActivity decreased 0/0 0/0 8/3 6/3Hypersensitive to touch 0/0 0/0 3/1 0/0ExcretionFeces discolored, Yellow 0/0 0/0 0/0 7/1Feces few/absent 0/0 5/2 2/2 0/0Feces watery 0/0 0/0 0/0 7/1Urine discolored, Brown 0/0 3/1 1/1 0/0Urine discolored, Orange 1/1 2/2 7/3 8/2Urine discolored, Red0/0 4/1 106/12 144/12External AppearanceLacrimation, Eye/right1/1 0/0 0/0 0/0Malocclusion 16/1 0/0 0/0 0/0Material around eyes, Red,Eye/right 16/1 0/0 0/0 0/0Material around mouth,Red 0/0 0/0 2/1 2/1Material around nose,Red 2/1 0/0 0/0 2/1Posture hunched0/0 5/2 9/3 8/1Swelling, Dorsal surface 0/0 0/0 11/1 0/0Thin 0/0 0/0 0/0 1/1Pelage/SkinHair discolored, Brown, Anogenital region 0/0 0/0 2/2 0/0Hair discolored, Yellow, Anogenital region 0/0 0/0 2/2 0/0Hair discolored, Yellow, Ventral surface 0/0 8/1 0/0 0/0Hair sparse, Abdominalregion 0/0 19/1 22/1 13/1Hair sparse, Hind limb/left 0/0 0/0 0/0 16/1Hair sparse, Hind limb/right 0/0 0/0 0/0 14/1 

# Page 42 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 2 Summary of P Premating/MatingClinical Findings+- FEMALE0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedPelage/SkinHair sparse, Inguinal region/right 0/00/0 0/0 17/1Hair sparse, Lumbar region 0/0 0/0 0/0 7/1Hair sparse, Ventral surface 0/0 12/1 0/0 3/1Piloerection 0/0 0/0 7/3 4/1Scabbed area, Dorsal surface 0/0 0/0 3/1 0/0Skin discolored, Pale, Entire body 0/0 0/0 0/0 1/1Unkempt appearance 0/0 0/0 0/0 2/1RespirationBreathing audible1/1 3/1 5/2 0/0Breathing rapid 0/0 0/0 2/1 0/0Rales 0/0 0/0 3/2 0/0 

# Page 43 of 358

# 1271-001 Table 3

Summary of P Gestatio n Clinical Findings Page 44 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 3 Summary of P GestationClinical Findings+0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedNumber of Animals Alive at Start of Interval11 11 10 10Behavior/ActivityActivity decreased 0/0 0/0 10/4 9/5ExcretionFeces discolored, Yellow 0/0 0/0 1/1 0/0Feces soft 0/0 0/0 0/0 12/1Feces watery 0/0 0/0 2/1 9/3Material in pan/bedding, Red 0/0 0/0 0/0 3/2Urine discolored, Orange 1/1 8/7 36/10 34/10Urine discolored, Red 0/0 48/11 66/10 71/10External AppearanceCarriage high 0/0 0/0 12/2 0/0Discharge, Clear, Anus 0/0 0/0 0/0 4/1Discharge, Red, Vulva 0/0 0/0 2/2 3/2Malocclusion 27/1 0/0 0/0 0/0Material around eyes, Red,Eye/right 14/1 0/0 0/0 0/0Material around mouth,Red 0/0 0/0 5/1 14/4Material around nose, Black 0/0 0/0 0/0 4/1Material around nose,Red 0/0 0/0 7/2 15/3Posture hunched0/0 3/1 0/0 3/1Thin 0/0 0/0 16/2 20/4Eye/OcularEye discolored, Pale, Eye/left 0/0 0/0 1/1 4/2Eye discolored, Pale, Eye/right 0/0 0/0 1/1 4/2Eyelid partially/completely closed, Eye/left 0/0 0/0 1/1 0/0Eyelid partially/completely closed, Eye/right 0/0 0/0 1/1 0/0 

# Page 45 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 3 Summary of P GestationClinical Findings+0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedPelage/SkinHair discolored, Brown, Anogenital region 0/0 6/1 18/2 2/1Hair discolored, Red, Abdominal region 0/0 0/0 2/1 1/1Hair discolored, Red, Anogenital region 0/0 0/0 19/5 1/1Hair discolored, Red, Ventral surface 0/0 10/1 0/0 0/0Hair discolored, Yellow, Anogenital region 0/0 0/0 1/1 0/0Hair discolored, Yellow, Ventral surface 0/0 10/1 0/0 0/0Hair sparse, Abdominalregion 0/0 28/1 37/2 6/1Hair sparse, Anogenital region 0/0 14/1 12/1 0/0Hair sparse, Cervical region 0/0 0/0 12/1 0/0Hair sparse, Forelimb/left 0/0 27/2 11/1 5/1Hair sparse, Forelimb/right 0/0 27/2 11/1 7/1Hair sparse, Hind limb/left 0/0 0/0 0/0 25/1Hair sparse, Hind limb/right 0/0 0/0 0/0 29/1Hair sparse, Lumbarregion 0/0 0/0 0/0 11/1Hair sparse, Ventral surface 0/0 27/1 0/0 11/1Piloerection 0/0 0/0 20/5 12/5Scabbed area, Dorsal surface 0/0 0/0 10/1 0/0Skin cold to touch 0/0 0/0 12/4 6/4Skin discolored, Blue, Abdominal region 0/0 0/0 0/0 1/1Skin discolored, Pale, Entire body 0/0 0/0 3/2 9/5RespirationBreathing shallow0/0 0/0 1/1 0/0Breathing slow 0/0 0/0 1/1 3/2Rales 0/0 1/1 0/0 0/0 

# Page 46 of 358

# 1271-001 Table 4

Summary of P Lactati on Clinical Findings Page 47 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 4 Summary of P LactationClinical Findings+0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedNumber of Animals Alive at Start of Interval11 10 9 4Behavior/ActivityActivity decreased 0/0 0/0 7/3 5/2Tremors 0/0 0/0 2/1 0/0ExcretionFeces watery 0/0 0/0 1/1 4/1External AppearanceCarriage high 0/0 0/0 11/3 0/0Discharge, Red, Vulva 0/0 0/0 2/2 0/0Malocclusion 5/1 0/0 0/0 0/0Material around eyes, Red,Eye/right 3/1 0/0 0/0 0/0Material around mouth,Red 0/0 0/0 2/1 2/1Material around nose,Red 0/0 0/0 4/2 2/1Thin 0/0 0/0 12/3 7/2Eye/OcularEye discolored, Pale, Eye/left 0/0 0/0 1/1 3/1Eye discolored, Pale, Eye/right 0/0 0/0 1/1 3/1Pelage/SkinHair discolored, Red, Abdominal region 0/0 0/0 2/1 0/0Hair discolored, Red, Anogenital region 0/0 0/0 4/3 0/0Hair sparse, Abdominal region 0/0 5/1 8/2 0/0Hair sparse, Anogenital region 0/0 5/1 4/1 0/0Hair sparse, Cervical region 0/0 0/0 4/1 0/0Hair sparse, Forelimb/left 0/0 5/1 5/1 0/0Hair sparse, Forelimb/right 0/0 5/1 5/1 0/0Hair sparse, Hind limb/left 0/0 0/0 0/0 2/1 

# Page 48 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 4 Summary of P LactationClinical Findings+0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayObservation+Number of times observed/Total number of animals affectedPelage/SkinHair sparse, Hind limb/right 0/0 0/0 0/0 5/1Hair sparse, Ventral surface 0/0 4/1 0/0 0/0Piloerection 0/0 0/0 8/3 6/2Skin cold to touch 0/0 0/0 11/5 2/1Skin discolored, Pale, Entire body 0/0 0/0 2/2 5/2RespirationBreathing shallow0/0 0/0 1/1 0/0Breathing slow 0/0 0/0 0/0 2/1 

# Page 49 of 358

# 1271-001 Table 5

Summary of P Body We ight Values - MALE Page 50 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 5 Summary of P Body Weight Values - MALEStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Week) Mean SD N Mean SD N Mean SD N Mean SD NBody Weight ValuesgN - Number of measures used to calculate meanSD - Standard DeviationPremating 1 260.39.33 12 265.113.55 12 263.116.12 12 262.213.27 122 305.214.68 12 298.824.58 12 293.517.64 12 291.523.11 12Pairing 3 348.921.20 12 337.124.20 12 326.8a18.59 12 326.5a24.06 124 378.226.60 12 366.922.90 12 350.6a22.62 12 354.430.21 12Postpairing 5 408.833.95 12 393.520.33 12 380.3a23.85 12 379.9a33.28 126 444.338.93 12 426.221.30 11 410.2a20.87 12 411.4a34.54 127 469.341.65 12 440.023.08 10 415.9b24.37 11 424.8b32.53 11aSignificantly different from control; (p<0.05)bSignificantly different from control; (p<0.01) 

# Page 51 of 358

# 1271-001 Table 6

Summary of P Premating/Mating Body Weight Values - FEMALE Page 52 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 6 Summary of PrematingBody Weight Values - FEMALEStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Week) Mean SD N Mean SD N Mean SD N Mean SD NBody Weight ValuesgN - Number of measures used to calculate meanSD - Standard Deviation1 195.48.39 12 193.85.86 12 195.88.90 12 203.412.24 122 214.312.32 12 205.810.09 12 190.9b16.33 12 207.316.48 123 229.514.37 12 212.2b8.72 10 206.9b14.28 11 211.2b11.54 9bSignificantly different from control; (p<0.01) 

# Page 53 of 358

# 1271-001 Table 7

Summary of P Gestatio n Body Weight Values Page 54 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 7 Summary of P GestationBody Weight ValuesStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Day) Mean SD N Mean SD N Mean SD N Mean SD NBody Weight ValuesgN - Number of measures used to calculate meanSD - Standard Deviation0 248.916.66 11 218.2b15.79 10 209.7b24.17 10 228.0a11.54 107 288.820.75 11 252.9b12.47 10 233.9b24.71 10 250.8b13.39 1014 324.224.00 11 288.2b15.84 10 263.1b28.59 10 269.6b16.75 1020 405.032.59 11 347.5b25.23 10 303.3b49.05 10 304.1b46.28 9aSignificantly different from control; (p<0.05)bSignificantly different from control; (p<0.01) 

# Page 55 of 358

# 1271-001 Table 8

Summary of P Lactati on Body Weight Values Page 56 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 8 Summary of P LactationBody Weight ValuesStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Day) Mean SD N Mean SD N Mean SD N Mean SD NBody Weight ValuesgN - Number of measures used to calculate meanSD - Standard Deviation0 300.925.49 11 252.1b21.75 10 201.8b27.34 9 227.8b17.48 44 314.522.52 11 269.0b39.14 10 203.5b28.64 4 237.3b22.69 4bSignificantly different from control; (p<0.01) 

# Page 57 of 358

# 1271-001 Table 9

Summary of P Gestation B ody Weight Change Values Page 58 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 9 Summary of P GestationBody Weight Change ValuesStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Day) Mean SD N Mean SD N Mean SD N Mean SD NBody Weight Change ValuesgN - Number of measures used to calculate meanSD - Standard Deviation0-7 39.98.07 11 34.76.18 10 24.2b9.59 10 22.8b13.16 107-14 35.47.46 11 35.37.12 10 29.211.52 10 18.8b11.82 1014-20 80.811.69 11 59.313.54 10 40.2b25.42 10 31.4b36.77 90-20 156.121.64 11 129.314.66 10 93.6b33.12 10 75.4b42.39 9bSignificantly different from control; (p<0.01) 

# Page 59 of 358

# 1271-001 Table 10

Summary of P Lactation B ody Weight Change Values Page 60 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 10 Summary of P LactationBody Weight Change ValuesStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Day) Mean SD N Mean SD N Mean SD N Mean SD NBody Weight Change ValuesgN - Number of measures used to calculate meanSD - Standard Deviation0-4 13.512.76 11 16.927.65 10 7.820.42 4 9.514.71 4 

# Page 61 of 358

# 1271-001 Table 11

Summary of P Food Consumption Values – MALE Page 62 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 11 Summary of P Food Consumption Values - MALEStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Week) Mean SD N Mean SD N Mean SD N Mean SD NFood Consumption Valuesg/animal/dayN - Number of measures used to calculate meanSD - Standard DeviationPremating 1-2 26.52.11 12 22.4b3.52 12 22.5b1.53 12 22.5b2.01 122-3 27.52.44 12 25.42.72 12 25.31.96 11 26.52.34 12Postmating 5-6 28.53.44 12 27.01.45 11 25.8a1.63 12 28.82.45 126-7 28.13.16 12 25.3a1.46 11 24.9a2.97 12 26.82.60 12aSignificantly different from control; (p<0.05)bSignificantly different from control; (p<0.01)NA- Not Applicable/Not Available 

# Page 63 of 358

# 1271-001 Table 12

Summary of P Premating Food Consumption Values - FEMALE Page 64 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 12 Summary of P PrematingFood Consumption Values - FEMALEStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Week) Mean SD N Mean SD N Mean SD N Mean SD NFood Consumption Valuesg/animal/dayN - Number of measures used to calculate meanSD - Standard Deviation1-2 17.92.17 12 14.4b1.69 12 10.8b1.69 12 12.9b2.14 112-3 19.32.17 12 17.2a1.96 12 16.7b2.10 11 18.11.64 12aSignificantly different from control; (p<0.05)bSignificantly different from control; (p<0.01) 

# Page 65 of 358

# 1271-001 Table 13

Summary of P Gestation Food Consumption Values Page 66 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 13 Summary of P GestationFood Consumption ValuesStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Day) Mean SD N Mean SD N Mean SD N Mean SD NFood Consumption Valuesg/animal/dayN - Number of measures used to calculate meanSD - Standard Deviation0-7 24.92.83 11 20.4b1.35 10 19.1b2.59 10 20.0b2.08 107-14 25.63.36 11 21.6b2.65 10 19.4b3.02 10 20.1b1.59 1014-20 28.02.43 11 23.3a3.66 10 20.4b4.43 10 19.4b5.64 90-20 26.12.74 11 21.7b1.94 10 19.6b2.96 10 19.8b2.21 9aSignificantly different from control; (p<0.05)bSignificantly different from control; (p<0.01) 

# Page 67 of 358

# 1271-001 Table 14

Summary of P Lactation Food Consumption Values Page 68 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 14 Summary of P LactationFood Consumption ValuesStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayIntervalEndpoint (Day) Mean SD N Mean SD N Mean SD N Mean SD NFood Consumption Valuesg/animal/dayN - Number of measures used to calculate meanSD - Standard Deviation0-4 37.14.70 11 32.46.87 9 12.3b2.39 4 18.0b9.94 4bSignificantly different from control; (p<0.01) 

# Page 69 of 358

# 1271-001 Table 15

Summary of P Macroscopic Observations – MALE Page 70 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary of P Macroscopic Observations - MALETable 15Terminal0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverity DOS SNC DOS SNC DOS SNC DOS SNCNumber of Animals Examined0 12 1 11 0 12 0 12DOS - Died or euthanized on studySNC - Scheduled necropsy00000000all tissueswithin normal limits0120101112cavity, thoracicadhesion-mild000100skinabrasion-mild000010thyroid glanddiscoloration, red- mild001000 

# Page 71 of 358

# 1271-001 Table 16

Summary of P Macroscopic Observations – FEMALE Page 72 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary of P Macroscopic Observations - FEMALETable 16Terminal0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverity DOS SNC DOS SNC DOS SNC DOS SNCNumber of Animals Examined0 12 0 12 8 4 7 5DOS - Died or euthanized on studySNC - Scheduled necropsy2200100010142010100001all tissueswithin normal limits01201143animal/whole bodybody fat depleted- mild000001kidneysdiscoloration-mild000000liverdiscoloration, pink- mild000001lobulation, accentuated- moderate000000stomach, glandularfocus/foci, black- mild000000focus/foci, red000000-minimal000000-moderate000000stomach, nonglandularfocus/foci, red- mild000100stomach, pyloruserosion/ulcer-moderate000000 

# Page 73 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary of P Macroscopic Observations - FEMALETable 16Terminal0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverity DOS SNC DOS SNC DOS SNC DOS SNCNumber of Animals Examined0 12 0 12 8 4 7 5DOS - Died or euthanized on studySNC - Scheduled necropsy20101020101010thymus glanddiscoloration, red000000-mild000000-moderate000000small000000-mild000000-moderate000000thyroid glandpale-mild000000 

# Page 74 of 358

# 1271-001 Table 17

Summary of P Organ We ight Values – MALE Page 75 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary of P Organ Weight Values - MALETable 17Terminal0mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpoint Mean SD N Mean SD N Mean SD N Mean SD NBody weightg47443124492110426b2212430b3612Epididymidesg1.2210.100121.1960.126111.1490.068121.1790.05912Epididymides/BWt%0.25890.0254120.26880.0253100.27030.0214120.27600.028412Prostate glg0.7280.227120.6200.115110.5880.162120.5950.11512Prostate gl/BWt%0.15410.0478120.14150.0169100.13800.0381120.13830.024512Sem. ves.g1.8010.208121.8520.224111.8810.236121.9270.21612Sem. ves./BWt%0.38240.0534120.41290.0572100.44190.0557120.4521a0.072712Testesg3.3150.157113.4940.217113.5270.299123.5120.22312Testes/BWt%0.70690.0732110.78370.0487100.8314b0.1027120.8231b0.101712N - Number of measures used to calculate meanSD - Standard DeviationaSignificantly different from control; (p<0.05)bSignificantly different from control; (p<0.01) 

# Page 76 of 358

# 1271-001 Table 18

Summary of P Organ Wei ght Values – FEMALE Page 77 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary of P Organ Weight Values - FEMALETable 18Terminal0mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpoint Mean SD N Mean SD N Mean SD N Mean SD NBody weightg3022111261b3910200b284222b205Ovariesg0.1370.027110.1230.017100.1170.01940.091b0.0265Ovaries/BWt%0.04510.0070110.04790.0071100.0584a0.004840.04050.00875Uterus w/ cervixg0.8060.140110.6780.133100.564a0.14740.585a0.1685Uterus w/ cervix/BWt%0.26640.0426110.26160.0453100.27920.038340.26110.06645N - Number of measures used to calculate meanSD - Standard DeviationaSignificantly different from control; (p<0.05)bSignificantly different from control; (p<0.01) 

# Page 78 of 358

# 1271-001 Table 19

Summary of P Microscopic Observations – MALE Page 79 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary of P Microscopic Observations - MALETable 19Terminal0 mg/kg/day2400 mg/kg/dayTissueObservationSeverity DOS SNC DOS SNCNumber of Animals Examined0 12 0 12DOS - Died or euthanized on studySNC - Scheduled necropsy( ) - Number observedepididymides(0)(12)(0)(12)within normal limits012012prostate gland(0)(12)(0)(12)infiltration, lymphocytic- minimal0404within normal limits0808seminal vesicles(0)(12)(0)(12)within normal limits012012testes(0)(12)(0)(12)within normal limits012012 

# Page 80 of 358

# 1271-001 Table 20

Summary of P Microscopi c Observations – FEMALE Page 81 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary of P Microscopic Observations - FEMALETable 20Terminal0 mg/kg/day2400 mg/kg/dayTissueObservationSeverity DOS SNC DOS SNCNumber of Animals Examined0 12 7 5DOS - Died or euthanized on studySNC - Scheduled necropsy( ) - Number observedliver(0)(0)(0)(1)infiltration, mononuclear cell- minimal0001necrosis, focal- minimal0001vacuolation, periportal- moderate0001ovaries(0)(12)(7)(5)within normal limits01275stomach, glandular(0)(0)(5)(0)edema0020-minimal0010-mild0010erosion/ulcer-mild0030within normal limits0020uterus(0)(12)(7)(5)within normal limits01275 

# Page 82 of 358

# 1271-001 Table 21

Summary of Premating Estrous Cycling Page 83 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 21 Summary of P Premating Estrous Cycling0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpoint Mean SD N Mean SD N Mean SD N Mean SD NN - Number of measures used to calculate meanaSignificantly different from control; (p<0.05)SD - Standard DeviationbSignificantlydifferent from control;(p<0.01)No. - NumberMean Cycle Length (Days) 4.4 0.44 11 4.7 1.16 12 5.6a1.43 10 4.8 0.68 10No. of Cycles (Count) 3.6 0.50 11 2.5b1.24 12 2.2b0.79 10 2.7a0.67 10 

# Page 84 of 358

# 1271-001 Table 22

Summary of Reproductive and Fertility Parameters Page 85 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 22 Summary of P Reproductive and Fertility Parameters0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpointNo. - NumberNo. Females on Study 12 12 12 12No. Females Paired 12 12 12 12No. Females Mated 11 11 12 11No. Pregnant 11 10 12 11Female Mating Index 91.7 91.7 100.0 91.7Female Fertility Index91.7 83.3 100.0 91.7Female Fecundity Index100.0 90.9 100.0 100.0No. Males on Study 12 12 12 12No. Males Paired 12 12 1212No. Males Mated 11 11 12 11No. Males Impregnating a Female 11 10 12 11Male Mating Index 91.7 91.7 100.0 91.7Male Fertility Index91.7 83.3 100.0 91.7Male Fecundity Index100.0 90.9 100.0 100.0 

# Page 86 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 22 Summary of P Reproductive and Fertility Parameters0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpointN - Number of measures used to calculate meanSD - Standard DeviationFemales with Confirmed Mating Day 11 11 10 10Copulatory Interval (Days)Mean 3.0 3.0 3.1 2.4SD 1.18 2.32 1.54 0.97N 11 11 8 10 

# Page 87 of 358

# 1271-001 Table 23

Summary of P Natural Delivery and Litter Data Page 88 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 23Summary of P Natural Delivery and Litter Data_________________________________________________________________________________________________________________________________SD - Standard Deviation1Not statistically analyzedaSignificantly different from control; (p<0.05)N - Number of measures used to calculate meanbSignificantly different from control; (p<0.01)No. - Number0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpointNo. Females on StudyN12121212No. Females PregnantN11101211Female Fertility Index%91.783.3100.091.7N111084Females Delivering Litters1%100.0100.066.736.4N0132With Stillborn Pups1%0.010.037.550.0N0010With All Stillborn1%0.00.012.50.0Mean22.122.623.1b23.0aSD0.300.520.690.82Gestation Length (Days)N111074Mean14.612.710.6a9.0aSD2.011.644.874.55No. of Pups at Day 0(Total Pups Born/Litter)N111084Mean14.612.68.1a7.8SD2.011.655.795.74Liveborn/LitterN111084 

# Page 89 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 23Summary of P Natural Delivery and Litter Data_________________________________________________________________________________________________________________________________SD - Standard DeviationaSignificantly different from control; (p<0.05)N - Number of measures used to calculate meanNo. - Number0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpointMean0.00.12.51.3SD0.000.324.601.50No. of Pups at Day 0 cont.Stillborn/LitterN111084N111074Gestation Index%100.0100.087.5100.0Mean %/Litter0.000.7719.9726.04SD0.0002.43336.01732.342Stillborn IndexN111084Mean15.513.913.0a12.5aSD1.441.202.882.08Total Implantation Scars/LitterN111084Mean %/Litter6.078.7920.4831.09aSD6.8516.84929.66728.177Post-Implantation LossN111084 

# Page 90 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 23Summary of P Natural Delivery and Litter Data_________________________________________________________________________________________________________________________________SD - Standard DeviationNA - Not Applicable/Not AvailablebSignificantly different from control; (p<0.01)N - Number of measures used to calculate meanNo. - Number0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayEndpointMean14.212.45.012.0SD1.831.945.66NANo. Live Pups/LitterDay 4N11921Mean %/Litter54.1745.0143.6533.97SD17.16113.47028.90739.239Sex Ratio (% Males per Animal)Pups Day 0N111053Mean %/Litter54.7245.5011.1125.00SD17.69814.64515.713NAPups Day 4N11921Mean %/Litter97.1989.0016.54b33.33bSD5.81831.42932.85057.735Pup Survival IndicesViability IndexN111053 

# Page 91 of 358

# 1271-001 Table 24

Summary of F 1 

# Pup Clinical Findings Page 92 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummaryof F1PupClinical Findings+Table 240 mg/kg/day 150 mg/kg/day 600 mg/kg/day 2400 mg/kg/dayObservation+ - Number of times observed/Total number of animals affectedNumber of Animals Alive at Start of Interval160 126 59 25Animals with No Abnormalities Detected160 123 25 25Behavior/ActivityActivity decreased 0/0 0/0 1/1 0/0External AppearanceLimb function impaired,Hind limb/left 0/0 2/2 0/0 0/0Limb function impaired,Hind limb/right 0/0 2/2 0/0 0/0Thin 0/0 0/0 1/1 0/0Pelage/SkinAbrasion(s), Cranial region 0/0 1/1 0/0 0/0Skin cold to touch 0/0 1/1 33/33 0/0Skin discolored, Pale, Entire body 0/0 1/1 1/1 0/0 

# Page 93 of 358

# 1271-001 Table 25

Summary of F 1 

# Pup Body Weight Values Page 94 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsTable 25 Summary of F1Pup Body Weight Values, gStudy0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayInterval SexSD - Standard Deviation ( ) - Least square meanbSignificantly different from control; (p<0.01)N - Number of measures used to calculate meanDay 0Males Mean 6.75 (6.82)6.60 (6.57) 5.06 (4.96)b6.60 (6.56)SD 0.466 0.576 0.788 0.000N 11 10 5 2Females Mean 6.45 (6.60)6.32 (6.30) 5.02 (4.79)b5.98 (5.93)SD 0.403 0.574 1.017 0.110N 11 10 6 2Males + Females Mean 6.60 (6.75) 6.46 (6.43) 5.17 (4.95)b6.32 (6.27)SD 0.443 0.551 0.988 0.175N 11 10 6 2Day 4Males Mean 9.74 (9.76) 8.75 (8.72) 4.80 (4.81)8.70 (8.66)SD 0.803 1.242 NA NAN 11 9 1 1Females Mean 9.29 (9.29) 8.53 (8.53) 3.93 (3.93)8.37 (8.38)SD 0.947 1.101 0.182 NAN 11 9 2 1Males + Females Mean 9.49 (9.50) 8.63 (8.63) 4.01 (4.01) 8.45 (8.44)SD 0.842 1.132 0.299 NAN 11 9 2 1 

# Page 95 of 358

# 1271-001 Table 26

Summary of F 1 

# Pup Macroscopic Observations Page 96 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary F1Pup Macroscopic Observations – MALETable 26StillbornPups0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverityNumber of Animals Examined00 103external appearancewithin normal limits00103 

# Page 97 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary F1Pup Macroscopic Observations – FEMALETable 26StillbornPups0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverityNumber of Animals Examined01 152external appearancewithin normal limits01152 

# Page 98 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary F1Pup Macroscopic Observations – MALETable 26Died on Study Pups0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverityNumber of Animals Examined26 235external appearancewithin normal limits26235 

# Page 99 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary F1Pup Macroscopic Observations – FEMALETable 26Died on Study Pups0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverityNumber of Animals Examined28 214external appearancewithin normal limits28214 

# Page 100 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary F1Pup Macroscopic Observations – MALETable 26Day4Pups0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverityNumber of Animals Examined87 5123external appearancewithin normal limits875123 

# Page 101 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsSummary F1Pup Macroscopic Observations – FEMALETable 26Day4Pups0 mg/kg/day150 mg/kg/day600 mg/kg/day2400 mg/kg/dayTissueObservationSeverityNumber of Animals Examined69 6189external appearancewithin normal limits696189 

# Page 102 of 358

# 1271-001 Appendix A

Vehicle and Test Article Information Page 103 of 358

# Vehicle and Test Article Information

Date Received: June 7, 2006

Supplier: Sigma Aldrich Inc.

St. Louis, Missouri

Amount Received: 1,000.0000 g

Label Identification: Guar (also known as Guar Gum)

Batch Number: 104K0080

Physical Characteristic s: Light white powder

Expiration Date: June 7, 2009

Storage: Room temperature

TMC Number: 06051B

Date Received: June 12, 2006

Supplier: Chemical Products Corporation

Cartersville, Georgia

Amount Received: 4,530.0000 g

Label Identification: Anthraquinone

Lot Number: 1543-1-GESU5099797

Physical Characteristics: Dark gray powder

Expiration Date: July 1, 2011

Storage: Room temperature, protected from light

TMC Number: 060534

Distilled water was provided by MPI Research, Inc.

1271-001 Page 104 of 358

# Page 105 of 358

# 1271-001 Appendix B

Analytical Report

(Sections 1 to 2) Page 106 of 358

# 1271-001 Appendix B

Section 1

Analytical Method Page 107 of 358

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# Page 115 of 358

# Page 116 of 358

# 1271-001 Appendix B

Section 2

Analysis of Dosing Formulations Page 117 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsHomogeneity Analysis of Dosing FormulationsRSD - Relative Standard Deviation1These data failed to meet acceptance criterion for RSD (≤10%).2These data failed to meet concentration acceptance criterion for recovery (±15%).Dose Level(mg/kg/day)Sample IdentificationNominalConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)AverageDose Level(mg/kg/day)%RecoveryMean%Recovery%RSD1501,2Test Batch Top Replicate 17.54.56374.166183.322060.855.526.5Test Batch Top Replicate 22.545733.9Test Batch Middle Replicate 15.589674.5Test Batch Middle Replicate 24.838564.5Test Batch Bottom Replicate 13.256943.4Test Batch Bottom Replicate 24.202456.024002Test Batch Top Replicate 1120.0079.136191.53631830.726065.976.37.2Test Batch Top Replicate 291.173376.0Test Batch Middle Replicate 198.498982.1Test Batch Middle Replicate 292.305376.9Test Batch Bottom Replicate 192.823577.4Test Batch Bottom Replicate 295.280679.4 

# Page 118 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsHomogeneity Analysis of Dosing FormulationsRSD - Relative Standard Deviation1These data failed to meet acceptance criterion for RSD (≤10%).2These data failed to meet concentration acceptance criterion for recovery (±15%).Dose Level(mg/kg/day)Sample IdentificationNominalConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)AverageDose Level(mg/kg/day)%RecoveryMean%Recovery%RSD1501,26/26/06 Top Replicate 17.53.47833.776575.530046.450.420.06/26/06 Top Replicate23.364844.96/26/06 Middle Replicate 14.796364.06/26/06 Middle Replicate 24.017253.66/26/06 Bottom Replicate 12.679335.76/26/06 Bottom Replicate 24.323257.624006/26/06 Top Replicate 1120.00113.9178104.96512099.302094.987.58.56/26/06 Top Replicate 2116.693397.26/26/06 Middle Replicate 1106.306788.66/26/06 Middle Replicate 299.657683.06/26/06 Bottom Replicate 194.490578.76/26/06 Bottom Replicate 298.724882.3 

# Page 119 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsWeek 3 Homogeneity Analysis of Dosing FormulationsRSD - Relative Standard Deviation1These data failed to meet acceptance criterion for RSD (≤10%).2These data failed to meet concentration acceptance criterion for recovery (±15%).Dose Level(mg/kg/day)Sample IdentificationNominalConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)AverageDose Level(mg/kg/day)%RecoveryMean%Recovery%RSD1501,2Top Replicate 17.55.45186.2297124.594072.783.112.0TopReplicate27.452199.4Middle Replicate 16.116981.6Middle Replicate 26.642488.6Bottom Replicate 15.531173.7Bottom Replicate 26.183982.524001Top Replicate 1120.00114.3310109.63682192.736095.391.413.7TopReplicate2118.422498.7Middle Replicate 1121.3300101.1Middle Replicate 279.888566.6Bottom Replicate 1112.481393.7Bottom Replicate 2111.367992.8 

# Page 120 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsStability Analysis of Dosing FormulationsRSD - Relative Standard Deviation1These data failed to meet acceptance criterion for recovery (±10%).2Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from the test batchhomogeneity analysis of dosing formulations.3Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from the 6/26/06homogeneity analysis of dosing formulations.StabilityInterval(Day)Dose Level(mg/kg/day)Sample IdentificationInitial CalculatedConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)%RecoveryMean%Recovery%RSD1215016/23/06 Replicate 15.214123.75713.447172.166.112.76/23/06 Replicate 23.137260.21224006/23/06 Replicate 195.4021292.996889.803797.594.15.06/23/06 Replicate 286.610690.8915016/26/06 Replicate 14.406833.52153.702179.984.06.96/26/06 Replicate 23.882888.19240016/26/06 Replicate 1102.9822399.275785.065296.482.623.66/26/06 Replicate 270.854868.8 

# Page 121 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsStability Analysis of Dosing FormulationsRSD - Relative Standard Deviation1These data failed to meet acceptance criterion for recovery (±10%).4Initial calculated concentration was basedon the average calculated concentration from the Week 3 concentration analyses of dosing formulations.5These data are the result of a redilutionand reanalysis conducted due to the original results failing to meet acceptance criterion for recovery (±10%).Reanalysis confirmed the originalresults; therefore, the meanof both analyses has been reported.StabilityInterval(Day)Dose Level(mg/kg/day)Sample IdentificationInitial CalculatedConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)%RecoveryMean%Recovery%RSD5715057/11/06 Replicate 15.812741.50391.721825.929.636.87/11/06 Replicate 21.590327.47/11/06 Redilution Replicate 11.162720.07/11/06 Redilution Replicate 22.630145.257240017/11/06 Replicate 1101.67584110.7990112.7601109.0110.92.57/11/06 Replicate 2114.7213112.8 

# Page 122 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsStability Analysis of Dosing FormulationsRSD - Relative Standard Deviation1These data failed to meet acceptance criterion for recovery (±10%).2Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from the test batchhomogeneity analysis of dosing formulations.3Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from the 6/26/06homogeneity analysis of dosing formulations.StabilityInterval(Day)Dose Level(mg/kg/day)Sample IdentificationInitial CalculatedConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)%RecoveryMean%Recovery%RSD211506/23/06 Replicate 15.214124.95984.821295.192.54.16/23/06 Replicate 24.682789.821240016/23/06 Replicate 195.4021276.608775.195480.378.82.76/23/06 Replicate 273.782077.31815016/26/06 Replicate 14.406835.083851.1182115.4116.11.06/26/06 Replicate 25.1526116.918240016/26/06 Replicate 1102.9822383.536483.665281.181.20.26/26/06 Replicate 283.794181.4 

# Page 123 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsStability Analysis of Dosing FormulationsRSD - Relative Standard Deviation1These data failed to meet acceptance criterion for recovery (±10%).4Initial calculated concentration was basedon the average calculated concentration from the Week 3 concentration analyses of dosing formulations.6These data are the result of a redilutionand reanalysis conducted due to the original results failing to meet acceptance criterion for recovery (±10%).StabilityInterval(Day)Dose Level(mg/kg/day)Sample IdentificationInitial CalculatedConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)%RecoveryMean%Recovery%RSD5115017/11/06 Replicate 15.812742.58762.789644.548.010.27/11/06 Replicate 22.991651.5571501,67/11/06 Redilution Replicate 15.812741.10751.730619.129.850.97/11/06 Redilution Replicate 22.353740.551240017/11/06 Replicate 1101.67584116.6508115.8385114.7113.91.07/11/06 Replicate 2115.0262113.1 

# Page 124 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsWeek 1 Concentration Analysis of Dosing FormulationsRSD - Relative Standard DeviationNA - Not Applicable/Not Available1These data failed to meet acceptance criterion for recovery (±15%).Dose Level(mg/kg/day)Sample IdentificationNominalConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)AverageDose Level(mg/kg/day)%RecoveryAverage%Recovery%RSD0Replicate10.00.00000.00000.0000NANANAReplicate20.0000NA1501Replicate 17.505.74025.4747109.494076.573.06.9Replicate25.209269.56001Replicate 130.0019.676219.8070396.140065.666.00.9Replicate219.937866.524001Replicate 1120.0085.165483.04701660.940071.069.23.6Replicate280.928667.4 

# Page 125 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsWeek 3 Concentration Analysis of Dosing FormulationsRSD - Relative Standard DeviationNA - Not Applicable/Not Available1These data failed to meet acceptance criterion for recovery (±15%).2These data failed to meet acceptance criterion for recovery (±15%) and RSD (≤10%).Dose Level(mg/kg/day)Sample IdentificationNominalConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)AverageDose Level(mg/kg/day)%RecoveryAverage%Recovery%RSD0Replicate10.00.00000.00000.0000NANANAReplicate20.0000NA1501Replicate 17.505.82375.8127116.254077.677.50.3Replicate25.801877.46002Replicate 130.0015.434217.8468356.936051.459.519.1Replicate220.259467.524002Replicate 1120.00109.6681101.67582033.516091.484.711.1Replicate293.683478.1 

# Page 126 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsWeek 8 Concentration Analysis of Dosing FormulationsRSD - Relative Standard DeviationNA - Not Applicable/Not Available1These data failed to meet acceptance criterion for recovery (±15%).3These data are the result of a reinjection conducted due to a failed check standard.4These data are the result of a redilutionand reanalysis conducted due to the original results failing to meet acceptance criteria for recovery (±15%) andRSD (≤10%). Reanalysis confirmed the original results; therefore, the mean of bothanalyses has been reported.Dose Level(mg/kg/day)Sample IdentificationNominalConcentration(mg/mL)CalculatedConcentration(mg/mL)AverageCalculatedConcentration(mg/mL)AverageDose Level(mg/kg/day)%RecoveryAverage%Recovery%RSD03Reinjection Replicate 10.00.00000.00000.0000NANANAReinjection Replicate 20.0000NA1503,4Reinjection Replicate 17.502.41081.880137.602032.125.128.6Reinjection Replicate 22.071027.6Redilution Replicate 11.139915.2Redilution Replicate 21.898925.36001,3Reinjection Replicate 130.0023.211223.1492462.984077.477.20.4Reinjection Replicate 223.087177.024003Reinjection Replicate 1120.00117.5559119.40882388.176098.099.52.2Reinjection Replicate 2121.2618101.1 

# Page 127 of 358

# 1271-001 Appendix C

Record of Animal Fate and Disposition Page 128 of 358

# MPI Research Study Number 1271-001

An Oral Reproduction/Developmental Toxicity Scre ening Study of Anthraquinone (CAS # 84-65-1) in Rats

Record of P Animal Fate and Disposition - MALE

Grou p

, Animal Number Fate Day

0 mg/kg/day

301 terminal necropsy 48

302 terminal necropsy 48

303 terminal necropsy 48

304 terminal necropsy 48

305 terminal necropsy 48

306 terminal necropsy 48

307 terminal necropsy 48

308 terminal necropsy 48

309 terminal necropsy 48

310 terminal necropsy 48

311 terminal necropsy 48

312 terminal necropsy 48

150 mg/kg/day

313 terminal necropsy 48

314 terminal necropsy 48

315 terminal necropsy 48

316 terminal necropsy 48

317 found dead 34

318 terminal necropsy 48

319 terminal necropsy 48

320 terminal necropsy 48

321 terminal necropsy 48

322 terminal necropsy 48

323 terminal necropsy 48

324 r 

# terminal necropsy 44

600 mg/kg/day

325 terminal necropsy 48

326 terminal necropsy 48

327 terminal necropsy 48

328 terminal necropsy 48

329 terminal necropsy 48

330 terminal necropsy 48

331 terminal necropsy 48

332 terminal necropsy 48

333 r 

# terminal necropsy 45

334 terminal necropsy 48

335 terminal necropsy 48

336 terminal necropsy 48

2400 mg/kg/day

337 r 

# terminal necropsy 44

338 terminal necropsy 48

339 terminal necropsy 48

340 terminal necropsy 48

r 

# Replacement animal 

# Page 129 of 358

# MPI Research Study Number 1271-001

An Oral Reproduction/Developmental Toxicity Scre ening Study of Anthraquinone (CAS # 84-65-1) in Rats

Record of P Animal Fate and Disposition - MALE

Grou p

, Animal Number Fate Day

2400 mg/kg/day

341 terminal necropsy 48

342 terminal necropsy 48

343 terminal necropsy 48

344 terminal necropsy 48

345 terminal necropsy 48

346 terminal necropsy 48

347 terminal necropsy 48

348 terminal necropsy 48 

# Page 130 of 358

# MPI Research Study Number 1271-001

An Oral Reproduction/Developmental Toxicity Scre ening Study of Anthraquinone (CAS # 84-65-1) in Rats

Record of P Animal Fate and Disposition - FEMALE

Grou p

, Animal Number Fate Day

0 mg/kg/day

349 terminal necropsy 51

350 terminal necropsy 48

351 terminal necropsy 48

352 terminal necropsy 55

353 terminal necropsy 49

354 terminal necropsy 50

355 terminal necropsy 51

356 terminal necropsy 50

357 terminal necropsy 48

358 terminal necropsy 49

359 terminal necropsy 47

360 terminal necropsy 49

150 mg/kg/day

361 terminal necropsy 50

362 terminal necropsy 47

363 terminal necropsy 55

364 terminal necropsy 48

365 terminal necropsy 51

366 terminal necropsy 51

367 terminal necropsy 51

368 terminal necropsy 48

369 r 

# terminal necropsy 44

370 terminal necropsy 48

371 r 

# terminal necropsy 50

372 terminal necropsy 55

600 mg/kg/day

373 r 

# terminal necropsy 44

374 found dead 46

375 found dead 47

376 found dead 45

377 terminal necropsy 51

378 r 

# found dead 41

379 found dead 44

380 found dead 48

381 r 

# found dead 49

382 euthanized in extremis 46

383 terminal necropsy 50

384 terminal necropsy 49

2400 mg/kg/day

385 found dead 54

386 found dead 44

387 terminal necropsy 49

388 found dead 45

r 

# Replacement animal 

# Page 131 of 358

# MPI Research Study Number 1271-001

An Oral Reproduction/Developmental Toxicity Scre ening Study of Anthraquinone (CAS # 84-65-1) in Rats

Record of P Animal Fate and Disposition - FEMALE

Grou p

, Animal Number Fate Day

2400 mg/kg/day

389 terminal necropsy 49

390 terminal necropsy 50

391 r 

# found dead 41

392 r 

# terminal necropsy 46

393 euthanized in extremis 39

394 found dead 44

395 r 

# found dead 40

396 terminal necropsy 55

r 

# Replacement animal 

# Page 132 of 358

# 1271-001 Appendix D

Individual P Clinical Findings

On occasion, clinical findings may have been observed more than once during the interval

and were recorded accordingly. The individual clinical findings table of this appendix

reports the findings observed, not the number of times observed. Page 133 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Clinical Findings - MALEGroup,Animal Number Day Sign Present0 mg/kg/day301No Abnormalities Detected 1, 3-47Hair discolored, Yellow, Anogenital region 2302No Abnormalities Detected 1-21, 24-47Urine discolored, Orange 22-23303No Abnormalities Detected 1-22Hair sparse, Forefoot/left 23-47Hair sparse, Forefoot/right 23-47304No Abnormalities Detected 1-47305No Abnormalities Detected 1-47306No Abnormalities Detected 1-47307No Abnormalities Detected 1-47308No Abnormalities Detected 1-47309No Abnormalities Detected 1-47310No Abnormalities Detected 1-47311No Abnormalities Detected 1-47312No Abnormalities Detected 1-47150 mg/kg/day313No Abnormalities Detected 1-27, 34, 36-37, 40-47Material around nose, Red 38-39Urine discolored, Orange 30Urine discolored, Red 28-29, 31-33, 35 

# Page 134 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Clinical Findings - MALEGroup,Animal Number Day Sign Present150 mg/kg/day314No Abnormalities Detected 1-22Hair sparse, Forelimb/left 23-47Hair sparse, Forelimb/right 23-47Urine discolored, Orange 30Urine discolored, Red 28-29, 31-33, 35315No Abnormalities Detected 1-23, 27-28, 34, 36-47Urine discolored, Orange 24-26, 30Urine discolored, Red 29, 31-33, 35316No Abnormalities Detected 1-2, 14-23, 27, 34, 36Hair sparse, Forefoot/left 3-13Hair sparse, Forefoot/right 3-13Hair sparse, Forelimb/left 37-47Hair sparse, Forelimb/right 37-47Urine discolored, Orange 24-26, 30Urine discolored, Red 28-29, 31-33, 35317No Abnormalities Detected 1-24, 27-28Urine discolored, Orange 25-26, 30Urine discolored, Red 29, 31-33318No Abnormalities Detected 1-22, 27-28, 34, 36-47Urine discolored, Orange 23-26, 30Urine discolored, Red 29, 31-33, 35319No Abnormalities Detected 1-23, 27-28, 34, 36-41, 43-47Urine discolored, Orange 24-26, 30Urine discolored, Red 29, 31-33, 35, 42320No Abnormalities Detected 1-22, 27, 34, 36-41, 45-47 

# Page 135 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Clinical Findings - MALEGroup,Animal Number Day Sign Present150 mg/kg/dayUrine discolored, Orange 23-26, 30Urine discolored, Red 28-29, 31-33, 35, 42-44321No Abnormalities Detected 1-24, 27-30, 34-47Urine discolored, Orange 25-26Urine discolored, Red 31-33322No Abnormalities Detected 1-24, 27-28Hair discolored, Brown, Forelimb/left 33-47Hair discolored, Brown, Forelimb/right 33-47Urine discolored, Orange 25-26, 30Urine discolored, Red 29, 31-33323No Abnormalities Detected 1-2, 15-24, 27-28, 34, 36-39, 41-47Activity decreased 3-5Breathing difficult 3-5Carriage low 4-5Piloerection 3-14Posture hunched 3-11Urine discolored, Orange 25-26, 30Urine discolored, Red 29, 31-33, 35, 40324rNo Abnormalities Detected 1-18, 23-24, 30, 32Hair sparse, Face 33-43Hair sparse, Ventral surface 35-43Urine discolored, Orange 19-22, 26Urine discolored, Red 25, 27-29, 31, 38600 mg/kg/day325No Abnormalities Detected 1-10, 22-28, 34, 36-47rReplacement animal 

# Page 136 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Clinical Findings - MALEGroup,Animal Number Day Sign Present600 mg/kg/dayUrine discolored, Orange 30Urine discolored, Red 11-21, 29, 31-33, 35326No Abnormalities Detected 2-10, 17Abrasion(s), Cervical region 27-47Hair sparse, Cervical region 37-47Salivation 1Scabbed area, Thoracic region 20-26Urine discolored, Orange 23-26, 30Urine discolored, Red 11-16, 18-21, 29, 31-33, 35-42327No Abnormalities Detected 1-7, 27, 34, 36-39Malocclusion 40-47Material around eyes, Black, Eye/left 44-47Material around eyes, Black, Eye/right 44-47Material around eyes, Red, Eye/left 40-43Material around eyes, Red, Eye/right 40-43Urine discolored, Orange 22-26, 28, 30Urine discolored, Red 8-21, 29, 31-33, 35, 40328No Abnormalities Detected 1-10, 17-18, 27, 34, 36-37Hair sparse, Forelimb/right 38-47Urine discolored, Orange 22-26, 28, 30Urine discolored, Red 11-16, 19-21, 29, 31-33, 35329No Abnormalities Detected 1-10,17-18, 22-24, 27, 34, 36-47Urine discolored, Orange 25-26, 28, 30Urine discolored, Red 11-16, 19-21, 29, 31-33, 35330No Abnormalities Detected 1-8, 22-24, 27, 34, 36-46Urine discolored, Orange 25-26, 28, 30 

# Page 137 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Clinical Findings - MALEGroup,Animal Number Day Sign Present600 mg/kg/dayUrine discolored, Red 9-21, 29, 31-33, 35, 47331No Abnormalities Detected 1-8, 22-24, 27, 34, 36Hair sparse, Forelimb/right 37-47Urine discolored, Orange 25-26, 28, 30Urine discolored, Red 9-21, 29, 31-33, 35, 47332No Abnormalities Detected 1-8, 22-24, 34, 36-46Urine discolored, Orange 25-28, 30Urine discolored, Red 9-21, 29, 31-33, 35, 47333rNo Abnormalities Detected 1-5, 19-21, 24, 31, 33-43Urine discolored, Orange 22-23, 25, 27Urine discolored, Red 6-18, 26, 28-30, 32, 44334No Abnormalities Detected 1-5,7-8, 22-24, 27-29, 34, 36-47Urine discolored, Brown 6Urine discolored, Orange 25-26, 30Urine discolored, Red 9-21, 31-33, 35335No Abnormalities Detected 1-2, 7-10, 22-24, 27-28, 34, 36-47Urine discolored, Brown 3-6Urine discolored, Orange 25-26, 30Urine discolored, Red 11-21, 29, 31-33, 35336No Abnormalities Detected 1-8, 22-24, 27, 34, 36-47Urine discolored, Orange 25-26, 28, 30Urine discolored, Red 9-21, 29, 31-33, 352400 mg/kg/day337rNo Abnormalities Detected 1-4,13-14, 18-21, 23, 30, 32-39, 41-43rReplacement animal 

# Page 138 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Clinical Findings - MALEGroup,Animal Number Day Sign Present2400 mg/kg/dayUrine discolored, Orange 22, 24, 26, 40Urine discolored, Red 5-12, 15-17, 25, 27-29, 31338No Abnormalities Detected 1-8, 17-18, 22-25, 27, 34, 36-47Urine discolored, Orange 26, 28, 30Urine discolored, Red 9-16, 19-21, 29, 31-33, 35339No Abnormalities Detected 4-7, 22-24, 27, 34, 36-43, 45-47Abrasion(s), Lip 1Hair discolored, Brown, Anogenital region 2-3Urine discolored, Orange 25-26, 28, 30, 44Urine discolored, Red 8-21, 29, 31-33, 35340No Abnormalities Detected 1-10, 22-24, 27Hair discolored, Brown, Ventral surface 30-47Hair discolored, Yellow, Ventral surface 29Urine discolored, Orange 25-26, 28, 30, 44Urine discolored, Red 11-21, 29, 31-33, 35341No Abnormalities Detected 1-11, 22-24, 27-28, 34, 36Hair sparse, Forefoot/left 37-47Hair sparse, Forefoot/right 37-47Urine discolored, Orange 25-26, 30, 44Urine discolored, Red 12-21, 29, 31-33, 35342No Abnormalities Detected 1-8, 22-24, 27, 34, 36-38, 41-43, 45-47Urine discolored, Orange 25-26, 28, 30, 44Urine discolored, Red 9-21, 29, 31-33, 35, 39-40343No Abnormalities Detected 1-8, 22-24, 27, 34, 36-43, 45-47Urine discolored, Orange 25-26, 28, 30, 44Urine discolored, Red 9-21, 29, 31-33, 35 

# Page 139 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Clinical Findings - MALEGroup,Animal Number Day Sign Present2400 mg/kg/day344No Abnormalities Detected 1-8, 22-24, 27, 34, 36-43, 45-47Urine discolored, Orange 25-26, 28, 30, 44Urine discolored, Red 9-21, 29, 31-33, 35345No Abnormalities Detected 1-8, 17-18, 22-24, 27-28, 34, 36-43, 45-47Urine discolored, Orange 25-26, 30, 44Urine discolored, Red 9-16, 19-21, 29, 31-33, 35346No Abnormalities Detected 1-8,17, 22-24, 27-28, 34, 36-43, 45-47Urine discolored, Orange 25-26, 30, 44Urine discolored, Red 9-16, 18-21, 29, 31-33, 35347No Abnormalities Detected 2-8, 22-24, 27, 34, 36-43, 45-46Rales 1Urine discolored, Orange 25-26, 28, 30, 44Urine discolored, Red 9-21, 29, 31-33, 35, 47348No Abnormalities Detected 2-4,6-8, 17, 22-24, 27, 34, 36Material around nose, Black 5Swelling, Anogenital region 1, 37-47Urine discolored, Orange 25-26, 28, 30, 44Urine discolored, Red 9-16, 18-21, 29, 31-33, 35 

# Page 140 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingClinical Findings - FEMALEGroup,Animal Number Day Sign Present0 mg/kg/day349No Abnormalities Detected 1-24350No Abnormalities Detected 1-21351No Abnormalities Detected 1-21352No Abnormalities Detected 1-33353No Abnormalities Detected 1-5, 7-22Breathing audible 6354No Abnormalities Detected 1-23355No Abnormalities Detected 1-23356No Abnormalities Detected 1-23357No Abnormalities Detected 1-21358No Abnormalities Detected 1-22359No Abnormalities Detected 1-20360No Abnormalities Detected 1-6Lacrimation, Eye/right 18Malocclusion 7-22Material around eyes, Red, Eye/right 7-22Material around nose, Red 7-8Urine discolored, Orange 22150 mg/kg/day361No Abnormalities Detected 1-22 

# Page 141 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingClinical Findings - FEMALEGroup,Animal Number Day Sign Present150 mg/kg/day362No Abnormalities Detected 1-4, 7-19Breathing audible 5-6Feces few/absent 20Posture hunched 20363No Abnormalities Detected 1-20Hair discolored, Yellow, Ventral surface 21-28Urine discolored, Orange 26364No Abnormalities Detected 1-20365No Abnormalities Detected 1-23366No Abnormalities Detected 1-4Hair sparse, Abdominal region 5-23367No Abnormalities Detected 1-3, 7-23Urine discolored, Brown 4-6368No Abnormalities Detected 1-21369rNo Abnormalities Detected 1-4Feces few/absent 5-8Hair sparse, Ventral surface 5-16Posture hunched 5-8370No Abnormalities Detected 1-21371rNo Abnormalities Detected 1-16372No Abnormalities Detected 1-28Urine discolored, Orange 30Urine discolored, Red 29, 31-33rReplacement animal 

# Page 142 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingClinical Findings - FEMALEGroup,Animal Number Day Sign Present600 mg/kg/day373rNo Abnormalities Detected 1-3Feces few/absent 4Piloerection 4-5Posture hunched 4-6Urine discolored, Red 7-15374No Abnormalities Detected 1-2, 7-10, 22Activity decreased 3-6Piloerection 3-6Urine discolored, Brown 6Urine discolored, Red 11-21375No Abnormalities Detected 1-3, 5-10Breathing audible 16Rales 4Urine discolored, Red 11-21376No Abnormalities Detected 1-5, 7-10Activity decreased 6Hair discolored, Yellow, Anogenital region 6Piloerection 6Urine discolored, Red 11-21377No Abnormalities Detected 1Activity decreased 3-5Breathing rapid 4-5Hair discolored, Brown, Anogenital region 4Hair sparse, Abdominal region 2-23Hypersensitive to touch 5-7Posture hunched 3-5rReplacement animal 

# Page 143 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingClinical Findings - FEMALEGroup,Animal Number Day Sign Present600 mg/kg/dayUrine discolored, Red 15-16, 18-21378rNo Abnormalities Detected 1-7, 12, 17Hair discolored, Brown, Anogenital region 18Urine discolored, Red 8-11, 13-16379No Abnormalities Detected 1-10, 17, 22-25, 27Urine discolored, Orange 26, 28, 30Urine discolored, Red 11-16, 18-21, 29, 31-33380No Abnormalities Detected 1-11, 17-18, 22-24, 27-28Urine discolored, Orange 25-26, 30Urine discolored, Red 12-16, 19-21, 29, 31-33381rNo Abnormalities Detected 1-2, 5-16, 19-21Rales 3-4Urine discolored, Orange 22Urine discolored, Red 17-18382No Abnormalities Detected 1-7, 11-13, 17-18Feces few/absent 9Posture hunched 8-10Urine discolored, Red 14-16, 19-21383No Abnormalities Detected 1, 3, 7-10, 17, 22-23Breathing audible 4-6Hair discolored, Yellow, Anogenital region 2Material around mouth, Red 4-5Urine discolored, Red 11-16, 18-21384No Abnormalities Detected 1-7Scabbed area, Dorsal surface 19-21rReplacement animal 

# Page 144 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingClinical Findings - FEMALEGroup,Animal Number Day Sign Present600 mg/kg/daySwelling, Dorsal surface 8-18Urine discolored, Red 15-16, 19-212400 mg/kg/day385No Abnormalities Detected 1-2, 7-10, 17, 22-24Activity decreased 3-6Feces discolored, Yellow 27-33Feces watery 27-33Material around mouth, Red 27Material around nose, Red 27Piloerection 30-33Posture hunched 27-33Skin discolored, Pale, Entire body 33Thin 33Unkempt appearance 32-33Urine discolored, Orange 25-26, 30Urine discolored, Red 11-16, 18-21, 29, 31-33386No Abnormalities Detected 1-11Urine discolored, Red 12-21387No Abnormalities Detected 1-8Urine discolored, Red 9-21388No Abnormalities Detected 1-8, 22Urine discolored, Red 9-21389No Abnormalities Detected 1Activity decreased 3Hair sparse, Abdominal region 6-18 

# Page 145 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingClinical Findings - FEMALEGroup,Animal Number Day Sign Present2400 mg/kg/dayHair sparse, Hind limb/left 6-21Hair sparse, Inguinal region/right 2-18Hair sparse, Lumbar region 15-21Hair sparse, Ventral surface 19-21Urine discolored, Red 9-21390No Abnormalities Detected 1-4, 6-8, 17, 22-23Activity decreased 5Urine discolored, Red 9-16, 18-21391rNo Abnormalities Detected 1-6, 13Urine discolored, Red 7-12, 14-17392rNo Abnormalities Detected 1-3Hair sparse, Hind limb/right 4-17Urine discolored, Red 5-12, 17393No Abnormalities Detected 1-8, 17Urine discolored, Red 9-16, 18-21394No Abnormalities Detected 1-8, 17-18, 22-24, 27Urine discolored, Orange 25-26, 28, 30Urine discolored, Red 9-16, 19-21, 29, 31-33395rNo Abnormalities Detected 1-4, 13Urine discolored, Red 5-12, 14-16396No Abnormalities Detected 1-8, 17, 22-23Urine discolored, Red 9-16, 18-21rReplacement animal 

# Page 146 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present0 mg/kg/day349No Abnormalities Detected 0-21350No Abnormalities Detected 0-21351No Abnormalities Detected 0-21353No Abnormalities Detected 0-21354No Abnormalities Detected 0-21355No Abnormalities Detected 0-22356No Abnormalities Detected 0-21357No Abnormalities Detected 0-21358No Abnormalities Detected 0-21359No Abnormalities Detected 0-21360Malocclusion 0-21Material around eyes, Red, Eye/right 0, 5-14Urine discolored, Orange 0150 mg/kg/day361No Abnormalities Detected 0-8, 11-13Hair sparse, Anogenital region 14-22Rales 16Urine discolored, Red 9-10362No Abnormalities Detected 7-10, 13, 17-21Hair discolored, Brown, Anogenital region 1-6Posture hunched 0-2 

# Page 147 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present150 mg/kg/dayUrine discolored, Orange 1-2Urine discolored, Red 11-12, 14-16363No Abnormalities Detected 20-21Hair discolored, Red, Ventral surface 10-19Hair discolored, Yellow, Ventral surface 0-9Urine discolored, Red 3-4, 6-8364No Abnormalities Detected 0-10, 13, 15-22Urine discolored, Red 11-12, 14365No Abnormalities Detected 0-7, 10, 12-22Urine discolored, Red 8-9, 11366Hair sparse, Abdominal region 0-22Urine discolored, Orange 6Urine discolored, Red 7-9, 11, 13367No Abnormalities Detected 0-5, 10, 12-22Urine discolored, Orange 6Urine discolored, Red 7-9, 11368No Abnormalities Detected 0-7, 12, 14-21Urine discolored, Orange 8Urine discolored, Red 9-11, 13369rHair sparse, Ventral surface 0-22Urine discolored, Orange 9Urine discolored, Red 10-12, 14-16370No Abnormalities Detected 0-7, 12, 14, 16Hair sparse, Forelimb/left 17-21Hair sparse, Forelimb/right 17-21rReplacement animal 

# Page 148 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present150 mg/kg/dayUrine discolored, Orange 8Urine discolored, Red 9-11, 13, 15371rNo Abnormalities Detected 0-5, 12, 14-17Hair sparse, Forelimb/left 18-24Hair sparse, Forelimb/right 18-24Urine discolored, Orange 8Urine discolored, Red 6-7, 9-11, 13600 mg/kg/day373rNo Abnormalities Detected 1-3, 6-7, 13-15Activity decreased 23Carriage high 16-17, 22-23Piloerection 16-23Skin cold to touch 23Thin 17-23Urine discolored, Orange 4-5, 9Urine discolored, Red 0, 8, 10-12, 16-19374No Abnormalities Detected 0-1, 4-5, 11, 13-21Activity decreased 22Breathing slow 22Piloerection 22Urine discolored, Orange 2-3, 7Urine discolored, Red 6, 8-10, 12375No Abnormalities Detected 0-2, 12, 14, 19-22Piloerection 23Skin cold to touch 23Skin discolored, Pale, Entire body 23rReplacement animal 

# Page 149 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present600 mg/kg/dayUrine discolored, Orange 3-6, 8Urine discolored, Red 7, 9-11, 13, 15-18376No Abnormalities Detected 0-2, 5-6, 12, 14-16, 19-21Feces discolored, Yellow 22Feces watery 22Skin cold to touch 22Urine discolored, Orange 3-4, 8Urine discolored, Red 7, 9-11, 13, 17-18377Hair sparse, Abdominal region 0-22Urine discolored, Orange 1-2, 4, 6Urine discolored, Red 5, 7-9, 11-13378rHair discolored, Brown, Anogenital region 0-14, 19Hair discolored, Red, Anogenital region 15-18, 21Hair discolored, Yellow, Anogenital region 20Piloerection 21Urine discolored, Orange 2, 4, 6Urine discolored, Red 5, 7-13381rNo Abnormalities Detected 1, 8, 10Hair discolored, Brown, Anogenital region 11-12Hair discolored, Red, Anogenital region 13-22Urine discolored, Orange 0, 2, 4Urine discolored, Red 3, 5-7, 9382No Abnormalities Detected 0-2, 5, 12, 14-19Activity decreased 22Eyelid partially/completely closed, Eye/left 22Eyelid partially/completely closed, Eye/right 22rReplacement animal 

# Page 150 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present600 mg/kg/dayMaterial around mouth, Red 20-22Material around nose, Red 20-22Piloerection 22Skin cold to touch 22Urine discolored, Orange 3-4, 6, 8Urine discolored, Red 7, 9-11, 13383No Abnormalities Detected 0, 3, 10, 12-21Urine discolored, Orange 1-2, 4, 6Urine discolored, Red 5, 7-9, 11384No Abnormalities Detected 12, 14Hair sparse, Abdominal region 17-22Hair sparse, Anogenital region 15-22Hair sparse, Cervical region 15-22Hair sparse, Forelimb/left 17-22Hair sparse, Forelimb/right 17-22Scabbed area, Dorsal surface 0-9Urine discolored, Orange 3-4, 6, 8Urine discolored, Red 7, 9-11, 13, 152400 mg/kg/day386No Abnormalities Detected 0-2, 5, 12, 14Hair sparse, Forelimb/left 17-21Hair sparse, Forelimb/right 15-21Urine discolored, Orange 3-4, 6, 8Urine discolored, Red 7, 9-11, 13, 15, 18387No Abnormalities Detected 0-2, 5, 12, 14, 16, 19-22Urine discolored, Orange 3-4, 6, 8 

# Page 151 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present2400 mg/kg/dayUrine discolored, Red 7, 9-11, 13, 15, 17-18388No Abnormalities Detected 0-1, 4-5, 11, 13Activity decreased 21Breathing slow 21Discharge, Clear, Anus 14-17Feces watery 18-21Hair discolored, Brown, Anogenital region 20-21Piloerection 19-21Posture hunched 19-21Skin discolored, Pale, Entire body 20-21Thin 19-21Urine discolored, Orange 2-3, 7Urine discolored, Red 6, 8-10, 12, 14389Hair sparse, Abdominal region 15-20Hair sparse, Hind limb/left 0-22Hair sparse, Lumbar region 0-10Hair sparse, Ventral surface 0-10Material around mouth, Red 22Material around nose, Red 22Urine discolored, Orange 6, 8Urine discolored, Red 7, 9-11390No Abnormalities Detected 0, 3, 10, 12, 14, 17-21Urine discolored, Orange 1-2, 4, 6Urine discolored, Red 5, 7-9, 11, 13, 15-16391rNo Abnormalities Detected 0-2, 5, 12, 14, 16, 19-21Activity decreased 22Feces watery 22rReplacement animal 

# Page 152 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present2400 mg/kg/dayPiloerection 22Skin cold to touch 22Skin discolored, Blue, Abdominal region 22Urine discolored, Orange 3-4, 6, 8Urine discolored, Red 7, 9-11, 13, 15, 17-18392rHair sparse, Hind limb/right 0-23Urine discolored, Orange 3-4, 6, 8Urine discolored, Red 7, 9-11, 13, 15, 20393No Abnormalities Detected 0-2, 12, 14Activity decreased 17Discharge, Red, Vulva 17Hair discolored, Red, Abdominal region 17Hair discolored, Red, Anogenital region 17Material around mouth, Red 17Material around nose, Black 15-17Material in pan/bedding, Red 17Piloerection 17Skin cold to touch 17Skin discolored, Pale, Entire body 17Urine discolored, Orange 3-6, 8Urine discolored, Red 7, 9-11, 13395rNo Abnormalities Detected 1-6, 13, 15, 17, 20-22Discharge, Red, Vulva 23Eye discolored, Pale, Eye/left 23Eye discolored, Pale, Eye/right 23Material around mouth, Red 23Material around nose, Red 23Material in pan/bedding, Red 23rReplacement animal 

# Page 153 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationClinical FindingsGroup,Animal Number Gestation Day Sign Present2400 mg/kg/daySkin cold to touch 23Skin discolored, Pale, Entire body 23Urine discolored, Orange 7, 9Urine discolored, Red0, 8, 10-12, 14, 16, 18-19396No Abnormalities Detected 0-3, 10, 12, 14Feces soft 21-31Material around mouth, Red 19-25, 31Material around nose, Red 17-25, 31Thin 23-31Urine discolored, Orange 4, 6Urine discolored, Red 5, 7-9, 11, 13, 15-16, 18 

# Page 154 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day349No Abnormalities Detected 0-4350No Abnormalities Detected 0-4351No Abnormalities Detected 0-4353No Abnormalities Detected 0-4354No Abnormalities Detected 0-4355No Abnormalities Detected 0-4356No Abnormalities Detected 0-4357No Abnormalities Detected 0-4358No Abnormalities Detected 0-4359No Abnormalities Detected 0-4360Malocclusion 0-4Material around eyes, Red, Eye/right 1-3150 mg/kg/day361Hair sparse, Anogenital region 0-4362No Abnormalities Detected 0-4363No Abnormalities Detected 0-4364No Abnormalities Detected 0-4365No Abnormalities Detected 0-4 

# Page 155 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day366Hair sparse, Abdominal region 0-4367No Abnormalities Detected 0-4368No Abnormalities Detected 0-4369rNo Abnormalities Detected 4Hair sparse, Ventral surface 0-3370Hair sparse, Forelimb/left 0-4Hair sparse, Forelimb/right 0-4600 mg/kg/day373rActivity decreased 0-3Carriage high 0-3Piloerection 0-4Skin cold to touch 0-3Thin 0-4375Activity decreased 0Discharge, Red, Vulva 0Eye discolored, Pale, Eye/left 0Eye discolored, Pale, Eye/right 0Hair discolored, Red, Anogenital region 0Piloerection 0Skin cold to touch 0Skin discolored, Pale, Entire body 0376Discharge, Red, Vulva 0Feces watery 0Hair discolored, Red, Anogenital region 0rReplacement animal 

# Page 156 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationClinical FindingsGroup,Animal Number Lactation Day Sign Present600 mg/kg/daySkin cold to touch 0Skin discolored, Pale, Entire body 0377No Abnormalities Detected 4Carriage high 0-3Hair sparse, Abdominal region 0-3Material around nose, Red 0-1Thin 0-3380Carriage high 0-2Skin cold to touch 0-2Thin 0-2Tremors 1-2381rNo Abnormalities Detected 0-2382Activity decreased 0-1Breathing shallow 1Hair discolored, Red, Abdominal region 0-1Hair discolored, Red, Anogenital region 0-1Material around mouth, Red 0-1Material around nose, Red 0-1Piloerection 0-1Skin cold to touch 0-1383No Abnormalities Detected 0-4384Hair sparse, Abdominal region 0-3Hair sparse, Anogenital region 0-3Hair sparse, Cervical region 0-3Hair sparse, Forelimb/left 0-4Hair sparse, Forelimb/right 0-4rReplacement animal 

# Page 157 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationClinical FindingsGroup,Animal Number Lactation Day Sign Present2400 mg/kg/day387No Abnormalities Detected 0-4389No Abnormalities Detected 1-4Activity decreased 0Breathing slow 0Hair sparse, Hind limb/left 0Material around mouth, Red 0Material around nose, Red 0Piloerection 0Skin cold to touch 0Skin discolored, Pale, Entire body 0390No Abnormalities Detected 0, 4Thin 1-3392rActivity decreased 0-2Eye discolored, Pale, Eye/left 0-2Eye discolored, Pale, Eye/right 0-2Feces watery 0-3Hair sparse, Hind limb/right 0-4Piloerection 0-3Skin discolored, Pale, Entire body 0-2Thin 0-3rReplacement animal 

# Page 158 of 358

# 1271-001 Appendix E

Individual P Body Weight Values Page 159 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Body Weight Values, g - MALEGroup,Animal Week of StudyNumber1 2 3 4 5 6 70 mg/kg/day301 273 324 372 407 451 501 529302 266 325 385 422 463 507 532303 263 295 331 356 383 409 433304 267 303 351 375 403 451 471305 260 290 324 347 352 383 415306 266 308 351 389 423 458 481307 247 299 332 359 388 415 435308 252 299 346 371 413 441 472309 271 325 367 407 424 462 497310 261 317 372 403 444 475 507311 244 281 320 344 368 400 411312 253 296 336 358 393 430 449150 mg/kg/day313 265 289 326 348 380 402 418314 262 305 341 365 399 431 444315 266 311 363 401 425 467 482316 267 306 350 384 414 445 471317 264 302 342 365 390 NDC318 254 290 337 355 389 430 437319 262 296 336 350 375 399 419320 257 289 323 354 376 402 409321 257 306 346 382 401 431 447322 257 300 333 364 372 426 443323 264 241 275 328 372 411 430324r306 351 373 407 429 444 NDCNDC- No data collected refer to record of faterReplacement animal 

# Page 160 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Body Weight Values, g - MALEGroup,Animal Week of StudyNumber1 2 3 4 5 6 7600 mg/kg/day325 258 289 322 342 360 395 410326 267 292 330 358 386 423 436327 255 286 328 349 383 408 370328 255 282 317 344 366 392 406329 250 285 319 343 383 414 431330 268 295 322 352 376 410 422331 269 298 328 345 376 407 405332 256 287 327 364 384 423 436333r308 344 376 401 432 449 NDC334 247 279 313 342 377 408 423335 270 305 341 365 407 428 453336 254 280 299 302 334 365 3832400 mg/kg/day337r300 355 382 416 452 475 NDC338 263 295 339 363 393 422 444339 263 282 314 340 375 404 423340 256 280 305 325 340 368 391341 266 295 336 382 402 440 457342 253 270 297 324 350 380 396343 254 272 316 326 355 381 401344 266 300 339 371 397 451 475345 265 299 327 363 387 416 429346 256 295 338 379 401 430 461347 248 267 293 314 331 363 372348 256 288 332 350 376 407 424NDC- No data collected refer to record of faterReplacement animal 

# Page 161 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingBody Weight Values, g - FEMALEGroup,Animal Week of StudyNumber1 2 3 4 50 mg/kg/day349 207 216 244 254350 187 200 217351 199 243 262352 184 202 214 233 249353 204 211 236 254354 193 212 219 234355 197 217 226 243356 207 219 230 250357 185 200 215358 202 226 238 255359 190 206 218360 190 220 235 263150 mg/kg/day361 197 215 217 228362 196 213 211363 199 202 216 219364 185 193 198365 192 203 216 223366 203 212 226 234367 200 225 218 232368 187 195 207369r188 212370 193 207 214371r198 202372 188 191 199 214 247rReplacement animal 

# Page 162 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Premating/MatingBody Weight Values, g - FEMALEGroup,Animal Week of StudyNumber1 2 3 4 5600 mg/kg/day373r178 162374 200 202 206 218375 192 181 190376 205 196 210377 195 186 191 185378r193 187 193379 202 190 214 212 233380 196 202 214 187 230381r203 224 236382 210 171 193383 187 204 219 228384 188 186 2102400 mg/kg/day385 198 203 206 215 186386 214 214 232387 213 215 216388 200 201 215 207389 188 188 209390 193 194 200 213391r226 235392r221 223393 198 188 199394 196 210 224 225 217395r203 229396 191 187 200 208rReplacement animal 

# Page 163 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationBody Weight Values, gGroup,Animal Day of GestationNumber0 7 14 200 mg/kg/day349 263 294 314 383350 228 265 307 388351 276 327 377 469353 259 297 335 434354 244 273 304 388355 248 281 316 388356 249 286 317 406357 231 275 313 380358 255 299 332 405359 222 262 296 364360 263 318 355 450 

# Page 164 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationBody Weight Values, gGroup,Animal Day of GestationNumber0 7 14 20150 mg/kg/day361 217 247 276 345362 186 230 270 319363 204 245 273 302364 211 250 271 339365 225 260 301 374366 232 263 297 350367 241 272 315 389368 212 244 283 340369r226 266 304 363370 228 252 292 354371rx NP 217 246 241 245600 mg/kg/day373r164 189 200 200374 PIDE 221 234 289 358375 197 234 265 316376 PIDE 214 250 279 330377 198 210 238 277378rDP 196 216 247 258381r248 260 288 313382 212 240 275 295383 242 274 291 366384 205 232 259 320DP- Died pregnantNP- Not pregnantPIDE- Pregnant, incomplete deliveryx- Excluded from summary statisticsrReplacement animal 

# Page 165 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationBody Weight Values, gGroup,Animal Day of GestationNumber0 7 14 202400 mg/kg/day386 DP 232 268 275 342387 243 256 282 296388 DP 215 259 254 229389 210 251 281 320390 226 240 266 337391rDP 242 262 298 352392r242 264 281 328393 PEM 222 226 242 NDC395rDP 224 241 261 305396 ALR 224 241 256 228ALR- All resorptionsDP- Died pregnantNDC- No data collected refer to record of fatePEM- Pregnant euthanized moribundrReplacement animal 

# Page 166 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationBody Weight Values, gGroup,Animal Day of LactationNumber0 40 mg/kg/day349 304 330350 283 301351 349 363353 323 318354 267 309355 283 296356 294 298357 289 291358 307 317359 277 295360 334 341150 mg/kg/day361 219 180362 264 252363 240 251364 227 273365 275 306366 240 273367 290 309368 247 258369r259 315370 260 273rReplacement animal 

# Page 167 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationBody Weight Values, gGroup,Animal Day of LactationNumber0 4600 mg/kg/day373r153 162374 PIDE NDC375 199 NDC376 PIDE 211 NDC377 180 209380 186 NDC381r228 NDC382 209 NDC383 247 227384 203 2162400 mg/kg/day387 229 221389 246 270390 232 235392r204 223NDC- No data collected refer to record of fatePIDE- Pregnant, incomplete deliveryrReplacement animal 

# Page 168 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual PostmatingBody Weight Values, g - FEMALEGroup,Animal Week of StudyNumber6 7 8NA- Not Applicable/Not Available0 mg/kg/day352 NP 253 252 265150 mg/kg/day371rNP NA 242 243372 NP 242 250 NDC600 mg/kg/day380 PNMD 264 306NDC- No data collected refer to record of fateNP- Not pregnantPNMD- Pregnant no mating daterReplacement animal 

# Page 169 of 358

# 1271-001 Appendix F

Individual P Body Weight Change Values Page 170 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationBody Weight Change Values, gGroup,Animal Day of GestationNumber0-7 7-14 14-20 0-200 mg/kg/day349 31 20 69 120350 37 42 81 160351 51 50 92 193353 38 38 99 175354 29 31 84 144355 33 35 72 140356 37 31 89 157357 44 38 67 149358 44 33 73 150359 40 34 68 142360 55 37 95 187 

# Page 171 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationBody Weight Change Values, gGroup,Animal Day of GestationNumber0-7 7-14 14-20 0-20150 mg/kg/day361 30 29 69 128362 44 40 49 133363 41 28 29 98364 39 21 68 128365 35 41 73 149366 31 34 53 118367 31 43 74 148368 32 39 57 128369r40 38 59 137370 24 40 62 126371rx NP 29 -5 4 28600 mg/kg/day373r25 11 0 36374 PIDE 13 55 69 137375 37 31 51 119376 PIDE 36 29 51 116377 12 28 39 79378rDP 20 31 11 62381r12 28 25 65382 28 35 20 83383 32 17 75 124384 27 27 61 115DP- Died pregnantNP- Not pregnantPIDE- Pregnant, incomplete deliveryx- Excluded from summary statisticsrReplacement animal 

# Page 172 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationBody Weight Change Values, gGroup,Animal Day of GestationNumber0-7 7-14 14-20 0-202400 mg/kg/day386 DP 36 7 67 110387 13 26 14 53388 DP 44 -5 -25 14389 41 30 39 110390 14 26 71 111391rDP 20 36 54 110392r22 17 47 86393 PEM 4 16 NDC395rDP 17 20 44 81396 ALR 17 15 -28 4ALR- All resorptionsDP- Died pregnantPEM- Pregnant euthanized moribundrReplacement animal 

# Page 173 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationBody Weight Change Values, gGroup,Animal Day of LactationNumber0-40 mg/kg/day349 26350 18351 14353 -5354 42355 13356 4357 2358 10359 18360 7150 mg/kg/day361 -39362 -12363 11364 46365 31366 33367 19368 11369r56370 13rReplacement animal 

# Page 174 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationBody Weight Change Values, gGroup,Animal Day of LactationNumber0-4600 mg/kg/day373r9377 29383 -20384 132400 mg/kg/day387 -8389 24390 3392r19rReplacement animal 

# Page 175 of 358

# 1271-001 Appendix G

Individual P Food Consumption Values Page 176 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Food Consumption Values, g/animal/day - MALEGroup,Animal Study Interval (Week)Number1-2 2-3 5-6 6-70 mg/kg/day301 29.028.632.131.7302 29.631.435.131.7303 24.024.424.726.0304 27.028.432.329.9305 25.324.027.024.7306 24.125.727.127.1307 26.426.725.726.1308 26.128.127.728.0309 29.129.329.631.6310 27.729.728.129.9311 23.124.423.121.4312 26.929.729.728.9150 mg/kg/day313 21.124.024.123.3314 24.927.127.725.9315 23.427.327.126.7316 23.726.628.725.6317 23.326.3NDC318 22.726.327.024.6319 22.023.925.922.9320 23.324.624.724.7321 24.327.127.428.0322 23.024.927.925.7323 11.918.328.125.4324r25.429.0427.926.04Food consumption calculated using 4 daysNDC- No data collected refer to record of faterReplacement animal 

# Page 177 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Food Consumption Values, g/animal/day - MALEGroup,Animal Study Interval (Week)Number1-2 2-3 5-6 6-7600 mg/kg/day325 23.627.025.025.4326 22.023.925.425.9327 22.427.724.917.1328 21.424.923.924.1329 21.125.427.626.3330 23.126.127.027.0331 24.425.025.025.0332 21.327.027.726.6333r25.625.5e26.925.6334 22.925.127.928.7335 22.725.625.425.7336 20.020.422.721.72400 mg/kg/day337r26.129.0430.726.6338 22.428.628.425.9339 22.323.927.423.3340 22.924.727.624.1341 21.728.728.028.7342 20.124.928.024.0343 21.627.132.130.9344 25.328.733.430.7345 23.126.727.626.0346 23.627.429.429.7347 18.721.424.025.4348 22.126.928.326.74Food consumption calculated using 4 dayseExcluded from the meanrReplacement animal 

# Page 178 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P PrematingFood Consumption Values, g/animal/day - FEMALEGroup,Animal Week of StudyNumber1-2 2-30 mg/kg/day349 18.7 20.1350 16.0 17.7351 21.3 23.3352 16.0 17.9353 18.0 19.1354 14.3 15.9355 19.3 19.1356 17.7 18.6357 15.3 18.1358 19.3 21.9359 17.9 18.1360 20.9 22.3150 mg/kg/day361 15.9 15.0362 16.3 17.0363 11.9 18.7364 13.9 15.0365 14.7 18.3366 15.1 17.1367 16.6 15.3368 13.4 17.1369r11.3 21.8370 15.7 18.7371r14.4 15.8372 13.3 16.4rReplacement animal 

# Page 179 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P PrematingFood Consumption Values, g/animal/day - FEMALEGroup,Animal Week of StudyNumber1-2 2-3600 mg/kg/day373r9.0 13.2374 10.9 15.4375 11.7 14.3376 10.0 19.1377 10.7 16.6378r13.0 16.0379 8.9 15.0380 13.0 18.3381r12.1382 7.7 19.9383 11.0 17.4384 12.1 18.12400 mg/kg/day385 11.1 19.4386 13.0 21.1387 13.1 17.0388 10.9 18.0389 12.0 18.6390 10.7 17.4391r15.1 19.0392r15.6 18.5393 39.6e16.1394 13.6 19.9395r16.6 15.5396 10.3 16.7eExcluded from the meanrReplacement animal 

# Page 180 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationFood Consumption Values, g/animal/dayGroup,Animal Day of GestationNumber0-7 7-14 14-20 0-200 mg/kg/day349 21.9 23.7 25.0 23.5350 22.3 23.4 27.2 24.2351 28.9 31.1 30.7 30.2353 25.1 26.3 31.8 27.6354 21.3 20.7 26.8 22.8355 24.7 23.6 26.7 24.9356 25.1 24.7 27.7 25.8357 24.9 24.6 26.2 25.2358 27.0 28.0 27.8 27.6359 22.4 24.1 26.2 24.2360 29.9 31.6 32.2 31.2 

# Page 181 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationFood Consumption Values, g/animal/dayGroup,Animal Day of GestationNumber0-7 7-14 14-20 0-20150 mg/kg/day361 18.9 18.9 22.7 20.0362 19.3 26.1 23.0 22.8363 19.1 20.9 14.0 18.2364 19.0 18.9 24.2 20.5365 22.7 23.9 27.5 24.6366 21.1 20.0 22.7 21.2367 22.1 23.7 26.5 24.0368 20.9 18.1 23.8 20.8369r20.4 22.4 23.3 22.0370 20.7 23.1 25.3 23.0371rx NP 19.7 16.4 15.5 17.3600 mg/kg/day373r15.1 14.9 11.8 14.1374 PIDE 18.3 22.6 25.2 21.9375 20.7 20.7 22.0 21.1376 PIDE 22.3 22.0 22.5 22.3377 15.4 14.6 18.0 15.9378rDP 18.4 16.7 15.2 16.9381r19.0 22.3 21.7 21.0382 18.9 20.9 18.8 19.6383 23.1 18.9 26.0 22.5384 20.0 20.7 22.7 21.1DP- Died pregnantNP- Not pregnantPIDE- Pregnant, incomplete deliveryx- Excluded from summary statisticsrReplacement animal 

# Page 182 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P GestationFood Consumption Values, g/animal/dayGroup,Animal Day of GestationNumber0-7 7-14 14-20 0-202400 mg/kg/day386 DP 22.4 19.7 23.2 21.7387 17.3 18.3 16.8 17.5388 DP 21.4 18.1 8.2 16.3389 21.3 20.4 20.0 20.6390 19.3 21.3 23.5 21.3391rDP 20.7 22.3 24.2 22.3392r22.6 19.7 22.0 21.4393 PEM 18.7 22.6 NDC395rDP 16.4 20.3 23.8 20.0396 ALR 19.7 18.4 13.0 17.3ALR- All resorptionsDP- Died pregnantNDC- No data collected refer to record of fatePEM- Pregnant euthanized moribundrReplacement animal 

# Page 183 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationFood Consumption Values, g/animal/dayGroup,Animal Day of LactationNumber0-40 mg/kg/day349 38.0350 32.8351 43.5353 30.8354 35.5355 39.8356 44.8357 39.8358 30.5359 36.5360 36.3150 mg/kg/day361 0.5e362 39.5363 23.8364 28.3365 41.3366 30.0367 35.5368 24.5369r40.3370 28.3eExcluded from the meanrReplacement animal 

# Page 184 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P LactationFood Consumption Values, g/animal/dayGroup,Animal Day of LactationNumber0-4600 mg/kg/day373r9.5377 15.0383 11.3384 13.32400 mg/kg/day387 17.5389 31.8390 8.3392r14.5rReplacement animal 

# Page 185 of 358

# 1271-001 Appendix H

Pathology Interpretation

Page 186 of 358

# 1271-001 MPI Research Study Number 1271-001

An Oral Reproduction/Developmental Toxici ty Screening Study of Anthraquinone

(CAS # 84-65-1) in Rats

Pathology Interpretation

Macroscopic

Test article-related macroscopic changes we re noted in female rats that received

Anthraquinone (CAS # 84-65-1), but not in males. Black or red foci were observed in the

glandular stomach of females at 600 and 2400 mg /kg/day that died on study (DOS) and an

erosion/ulcer was noted in the pyloric stomach of one DOS female at 2400 mg/kg/day. Black

or red foci and the erosion/ulcer corresponded microscopically with erosion/ulcer of the

glandular stomach.

Other macroscopic changes noted were typical of those commonly seen in rats of the same

strain and age or did not demonstrate a dose-re lated response, and were considered to be

incidental.

Organ Weights

No test article-related organ weight cha nges were noted in rats of either sex.

In males, the relative (to body weight) seminal vesicle weight at 2400 mg/kg/day and relative

(to body weight) left and right testes weights at 600 and 2400 mg/kg/day were statistically

significantly increased compared to the contro ls, but the relative increases were caused by

decreased body weight and were no t primary organ weight changes.

In females, statistically significant decreases were noted in the mean weight of the ovaries at

2400 mg/kg/day and the mean weight of the uterus at 600 and 2400 mg/kg/day. Decreased

weight of the ovaries and uterus was consider ed to be secondary to significantly decreased

body weights in these groups and not primary organ weight changes. 11

11 

# Levin S, Semler D, and Ruben Z. Effects of two weeks of feed restriction on some common toxicologic

parameters in Sprague-Dawley rats. Toxicologic Pathology, 21(1): 1-14, 1993. 

# Page 187 of 358

# Page 188 of 358

# 1271-001 Appendix I

Tissues and Organs Collected, Weighed, and Examined Page 189 of 358

# The following list constitutes the full complement of organs and tissues:

- Epididymis (2)*

- Gonads:

ovary (2)*

testis (2)* - Gross lesions

- Prostate* and seminal vesicle (2)*

- Uterus [both horns]/Cervix*

*Weighed organ

(2) Paired organ



# 1271-001 Page 190 of 358

# 1271-001 Appendix J

Individual P Organ Weight Values Page 191 of 358

# Codes For Individual Organ Weight Values

Abbreviations

BWt - Body Weight

gl - gland

lt - left

rt - right

Sem. - Seminal

ves. - vesicle

w/ - with

1271-001 Page 192 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalBodyweight Epididymides Epididymides/ Prostategl Prostategl/ Sem. ves. Sem. ves./ Testis, lt Testis, lt/Group,BWtBWtBWtBWtAnimalNumber g g % g % g % g %0 mg/kg/day3015141.1460.22300.4090.07961.6820.32721.5410.29983025531.1890.21500.8410.15212.1300.38521.6800.30383034341.2750.29381.0590.24401.6120.37141.5990.36843044691.1020.23500.6740.14371.6430.35031.5670.33413054181.0920.26120.5340.12781.9660.47031.6720.40003064891.2820.26220.4230.08651.6980.34724.025e0.8231e3074441.1720.26400.6320.14231.6480.37121.5940.35903084721.2990.27520.9510.20151.6620.35211.7050.36123095141.4450.28111.0710.20841.5490.30141.8060.35143105121.2170.23770.8530.16662.1220.41450.46611.7761.6710.34690.40173113124164541.1561.2770.27790.28130.6890.5960.16560.13131.9391.9630.43241.6610.3659eExcluded from the mean 

# Page 193 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalTestis, rt Testis, rt/Group,BWtAnimalNumber g %0 mg/kg/day3011.5120.29423021.6710.30223031.5420.35533041.5890.33883051.7610.42133061.9960.40823071.5800.35593081.7360.36783091.6540.32183101.7350.33893111.7480.42023121.6700.3678 

# Page 194 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalBodyweight Epididymides Epididymides/ Prostategl Prostategl/ Sem. ves. Sem. ves./ Testis, lt Testis, lt/Group,BWtBWtBWtBWtAnimalNumber g g % g % g % g %150 mg/kg/day313NA1.081NA0.436NA1.863NA1.602NA3144511.1520.25540.6200.13752.0570.45611.5760.34943154841.1220.23180.7310.15101.6910.34941.8240.37693164851.5180.31300.8550.17631.6760.34561.8810.38783184491.1680.26010.6100.13591.8840.41961.7960.40003194291.0470.24410.5440.12681.5330.35731.6320.38043204181.2490.29880.5300.12681.6610.39741.8190.43523214491.1260.25080.5750.12811.8310.40781.7750.39533224521.2460.27570.6740.14912.1820.48271.8470.40863234341.2400.28570.5430.12512.2370.51541.7670.4071r4431.2080.27270.7010.15821.7620.39771.6670.3763324NA- Not Applicable/Not AvailablerReplacement animal 

# Page 195 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalTestis, rt Testis, rt/Group,BWtAnimalNumber g %150 mg/kg/day3131.650NA3141.5650.34703151.7930.37053161.9510.40233181.7800.39643191.5910.37093201.8290.43763211.7900.39873221.7810.39403231.8320.4221r1.6870.3808324NA- Not Applicable/Not AvailablerReplacement animal 

# Page 196 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalBodyweight Epididymides Epididymides/ Prostategl Prostategl/ Sem. ves. Sem. ves./ Testis, lt Testis, lt/Group,BWtBWtBWtBWtAnimalNumber g g % g % g % g %600 mg/kg/day3254121.0100.24510.2980.07231.3760.33401.5710.38133264391.1260.25650.5220.11892.0940.47701.7070.38883274001.0690.26730.3520.08801.8860.47151.9320.48303284071.0970.26950.6740.16561.9210.47201.7660.43393294341.1580.26680.5830.13431.9100.44011.5920.36683304271.2250.28690.6200.14521.6950.39701.8530.43403314131.2120.29350.6760.16372.2270.53921.9940.48283324471.1340.25370.5950.13312.0580.46041.6600.3714r4471.1500.25730.4550.10181.6290.36441.6780.37543333344431.1910.26880.6560.14812.0950.47291.7510.39533354571.1690.25580.8650.18931.9180.41971.6230.35513363881.2510.32240.7570.19511.7620.45411.9130.4930rReplacement animal 

# Page 197 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalTestis, rt Testis, rt/Group,BWtAnimalNumber g %600 mg/kg/day3251.5490.37603261.6590.37793271.9940.49853281.7710.43513291.5120.34843301.8810.44053312.0090.48643321.7510.3917r1.7850.39933333341.7860.40323351.6470.36043361.9340.4985rReplacement animal 

# Page 198 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalBodyweight Epididymides Epididymides/ Prostategl Prostategl/ Sem. ves. Sem. ves./ Testis, lt Testis, lt/Group,BWtBWtBWtBWtAnimalNumber g g % g % g % g %2400 mg/kg/dayr3374691.2210.26030.8060.17191.9790.42201.8090.38573384471.1430.25570.5730.12821.9160.42861.7080.38213394291.1730.27340.5290.12332.1760.50721.7840.41593403931.2140.30890.5310.13512.1440.54551.6040.40813414621.2750.27600.5530.11971.6990.36771.8900.40913423901.1640.29850.4630.11871.9860.50921.7660.45283433881.1660.30050.6640.17112.0310.52351.8080.46603444781.0350.21650.5830.12201.9370.40521.6030.33543454391.2250.27900.8310.18932.1600.49201.7000.38723464631.1770.25420.5580.12051.6720.36111.5540.33563473751.2010.32030.4960.13231.9580.52211.9240.51313484291.1530.26880.5480.12771.4620.34081.8510.4315rReplacement animal 

# Page 199 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - MALETerminalTestis, rt Testis, rt/Group,BWtAnimalNumber g %2400 mg/kg/dayr3371.7690.37723381.6980.37993391.8400.42893401.6330.41553411.8550.40153421.7330.44443431.7800.45883441.6300.34103451.7100.38953461.6380.35383471.9690.52513481.8820.4387rReplacement animal 

# Page 200 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - FEMALETerminalBodyweight Ovaries Ovaries/BWt Uterus w/ Uterus w/Group,cervix cervix/BWtAnimalNumber g g % g %0 mg/kg/day3493190.1490.04670.9490.29753502880.1530.05311.0870.37743513480.1980.05690.9360.2690eeeee0.0980.03750.9770.37433522613533040.1200.03950.6980.22963542950.1240.04200.6670.22613552870.1390.04840.6820.23763562890.1350.04670.7620.26373572840.0900.03170.6860.24153583040.1180.03880.7600.25003593602803270.1320.1460.04710.04460.7300.9080.26070.2777eExcluded from the mean 

# Page 201 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - FEMALETerminalBodyweight Ovaries Ovaries/BWt Uterus w/ Uterus w/Group,cervix cervix/BWtAnimalNumber g g % g %150 mg/kg/day3611770.1020.05760.4810.27183622420.1250.05170.5300.21903632340.1220.05210.7790.33293642620.1250.04770.6770.25843652960.1040.03510.6400.21623662680.1530.05710.5280.19703673010.1250.04150.7390.24553682530.1050.04150.8330.3292r3090.1440.04660.8650.27993693702650.1270.04790.7050.2660reeeee2400.1290.05380.5000.2083371eeeee3722170.1160.05350.3030.1396eExcluded from the meanrReplacement animal 

# Page 202 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - FEMALETerminalBodyweight Ovaries Ovaries/BWt Uterus w/ Uterus w/Group,cervix cervix/BWtAnimalNumber g g % g %600 mg/kg/dayr3731600.0900.05630.3740.23383772040.1220.05980.5720.28043832240.1190.05310.7330.32723842100.1350.06430.5780.2752rReplacement animal 

# Page 203 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Organ Weight Values - FEMALETerminalBodyweight Ovaries Ovaries/BWt Uterus w/ Uterus w/Group,cervix cervix/BWtAnimalNumber g g % g %2400 mg/kg/day3872150.0760.03530.6410.29813892550.1180.04630.6440.25253902270.1170.05150.7910.3485r2140.0860.04020.5000.23363923962010.0590.02940.3470.1726rReplacement animal 

# Page 204 of 358

# 1271-001 Appendix K

Individual P Animal Data Record: Pathology Page 205 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study301SMacroscopicall tissues- within normal limits301SMicroscopicepididymides- within normal limitsprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits302SMacroscopicall tissues- within normal limits302SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits303SMacroscopicall tissues- within normal limits303SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits304SMacroscopicall tissues- within normal limits 

# Page 206 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study304SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits305SMacroscopicall tissues- within normal limits305SMicroscopicepididymides- within normal limitsprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits306SMacroscopicall tissues- within normal limits306SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits307SMacroscopicall tissues- within normal limits307SMicroscopicepididymides- within normal limits 

# Page 207 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study307 SMicroscopicprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits308SMacroscopicall tissues- within normal limits308SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits309SMacroscopicall tissues- within normal limits309SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits310SMacroscopicall tissues- within normal limits310SMicroscopicepididymides- within normal limitsprostate gland- within normal limits 

# Page 208 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study310 SMicroscopicseminal vesicles- within normal limitstestes- within normal limits311SMacroscopicall tissues- within normal limits311SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits312SMacroscopicall tissues- within normal limits312SMicroscopicepididymides- within normal limitsprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits 

# Page 209 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study313SMacroscopicall tissues- within normal limits314SMacroscopicall tissues- within normal limits315SMacroscopicall tissues- within normal limits316SMacroscopicall tissues- within normal limits317DMacroscopicthyroid gland- discoloration, red, bilateral, mild318SMacroscopicall tissues- within normal limits319SMacroscopicall tissues- within normal limits320SMacroscopicall tissues- within normal limits 

# Page 210 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations150 mg/kg/day321SMacroscopicall tissues- within normal limits322SMacroscopicall tissues- within normal limits323SMacroscopiccavity, thoracic- adhesion, mildall thoracic organs adhered to one another324rSMacroscopicall tissues- within normal limitsS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 211 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study325SMacroscopicall tissues- within normal limits326SMacroscopicskin- abrasion, right lateral neck, mildcorresponds to antemortem observation (abrasion(s); hair sparse)327SMacroscopicall tissues- within normal limits328SMacroscopicall tissues- within normal limits329SMacroscopicall tissues- within normal limits330SMacroscopicall tissues- within normal limits331SMacroscopicall tissues- within normal limits332SMacroscopicall tissues- within normal limits 

# Page 212 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations600 mg/kg/day333rSMacroscopicall tissues- within normal limits334SMacroscopicall tissues- within normal limits335SMacroscopicall tissues- within normal limits336SMacroscopicall tissues- within normal limitsS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 213 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/day337rSMacroscopicall tissues- within normal limits337SMicroscopicepididymides- within normal limitsprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits338SMacroscopicall tissues- within normal limits338SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits339SMacroscopicall tissues- within normal limits339SMicroscopicepididymides- within normal limitsprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits340SMacroscopicall tissues- within normal limitsS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 214 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study340SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits341SMacroscopicall tissues- within normal limits341SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits342SMacroscopicall tissues- within normal limits342SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits343SMacroscopicall tissues- within normal limits343SMicroscopicepididymides- within normal limits 

# Page 215 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study343SMicroscopicprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits344SMacroscopicall tissues- within normal limits344SMicroscopicepididymides- within normal limitsprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits345SMacroscopicall tissues- within normal limits345SMicroscopicepididymides- within normal limitsprostate gland- infiltration, lymphocytic, minimalseminal vesicles- within normal limitstestes- within normal limits346SMacroscopicall tissues- within normal limits346SMicroscopicepididymides- within normal limitsprostate gland- within normal limits 

# Page 216 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - MALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study346SMicroscopicseminal vesicles- within normal limitstestes- within normal limits347SMacroscopicall tissues- within normal limits347SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits348SMacroscopicall tissues- within normal limits348SMicroscopicepididymides- within normal limitsprostate gland- within normal limitsseminal vesicles- within normal limitstestes- within normal limits 

# Page 217 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study349SMacroscopicall tissues- within normal limits349SMicroscopicovaries- within normal limitsuterus- within normal limits350SMacroscopicall tissues- within normal limits350SMicroscopicovaries- within normal limitsuterus- within normal limits351SMacroscopicall tissues- within normal limits351SMicroscopicovaries- within normal limitsuterus- within normal limits352SMacroscopicall tissues- within normal limits352SMicroscopicovaries- within normal limitsuterus- within normal limits353SMacroscopicall tissues- within normal limits 

# Page 218 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study353SMicroscopicovaries- within normal limitsuterus- within normal limits354SMacroscopicall tissues- within normal limits354SMicroscopicovaries- within normal limitsuterus- within normal limits355SMacroscopicall tissues- within normal limits355SMicroscopicovaries- within normal limitsuterus- within normal limits356SMacroscopicall tissues- within normal limits356SMicroscopicovaries- within normal limitsuterus- within normal limits357SMacroscopicall tissues- within normal limits357SMicroscopicovaries- within normal limits 

# Page 219 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyMicroscopic357Suterus- within normal limits358SMacroscopicall tissues- within normal limits358SMicroscopicovaries- within normal limitsuterus- within normal limits359SMacroscopicall tissues- within normal limits359SMicroscopicovaries- within normal limitsuterus- within normal limits360SMacroscopicall tissues- within normal limits360SMicroscopicovaries- within normal limitsuterus- within normal limits 

# Page 220 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study361SMacroscopicstomach, nonglandular- focus/foci, red, focal, mild362SMacroscopicall tissues- within normal limits363SMacroscopicall tissues- within normal limits364SMacroscopicall tissues- within normal limits365SMacroscopicall tissues- within normal limits366SMacroscopicall tissues- within normal limits367SMacroscopicall tissues- within normal limits368SMacroscopicall tissues- within normal limits 

# Page 221 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations150 mg/kg/day369rSMacroscopicall tissues- within normal limits370SMacroscopicall tissues- within normal limits371rSMacroscopicall tissues- within normal limits372SMacroscopicall tissues- within normal limitsS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 222 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations600 mg/kg/day373rSMacroscopicall tissues- within normal limits374DMacroscopicstomach, glandular- focus/foci, red, mucosa, moderate375DMacroscopicall tissues- within normal limits376DMacroscopickidneys- discoloration,mottled, bilateral, mildstomach, glandular- focus/foci, black, mucosa, mildthyroid gland- pale, bilateral, milduterus with cervix- within normal limitsincomplete delivery. this animal had 14 implantation scars and onenon-viable fetus in utero. this fetus was externallynormal.377SMacroscopicall tissues- within normal limits378rDMacroscopicstomach, glandular- focus/foci, red, mucosa, minimal379DMacroscopicall tissues- within normal limitsS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 223 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations600 mg/kg/dayMacroscopic380Dthymus gland- discoloration, red, moderate- small, moderate381rMacroscopicDthymus gland- discoloration, red, mild- small, mild382MacroscopicEliver- lobulation, accentuated, multiple lobes, moderate383MacroscopicSall tissues- within normal limits384MacroscopicSall tissues- within normal limitsS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 224 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study385DMacroscopicstomach, glandular- focus/foci, black, multiple, mucosa, mild385DMicroscopicovaries- within normal limitsstomach, glandular- within normal limitsuterus- within normal limitsnon-correlated macro observation- stomach, glandular - focus/foci, black386DMacroscopicstomach, glandular- focus/foci, black, mucosa, mild386DMicroscopicovaries- within normal limitsstomach, glandular- edema, mild- erosion/ulcer, mildcorresponds to macroscopic observation (stomach, glandular -focus/foci, black)uterus- within normal limits387SMacroscopicall tissues- within normal limits387SMicroscopicovaries- within normal limitsuterus- within normal limits388DMacroscopicstomach, glandular- focus/foci, black, mucosa, mild 

# Page 225 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/day388DMicroscopicovaries- within normal limitsstomach, glandular- erosion/ulcer, mildcorresponds to macroscopic observation (stomach, glandular -focus/foci, black)uterus- within normal limits389SMacroscopicall tissues- within normal limits389SMicroscopicovaries- within normal limitsuterus- within normal limits390SMacroscopicliver- discoloration, pink, multifocal, multiple lobes, mild390SMicroscopicliver- infiltration, mononuclear cell, minimal- necrosis, focal, minimalcorresponds to macroscopic observation (liver - discoloration, pink)- vacuolation, periportal, hepatocytes, moderateovaries- within normal limitsuterus- within normal limits391rDMacroscopicstomach, glandular- focus/foci, black, mucosa, mildfoci located in glandular and pylorus portions of the stomach.S - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 226 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/day391DMicroscopicovaries- within normal limitsstomach, glandular- edema, minimal- erosion/ulcer, mildcorresponds to macroscopic observation (stomach, glandular -focus/foci, black)uterus- within normal limits392rSMacroscopicall tissues- within normal limits392SMicroscopicovaries- within normal limitsuterus- within normal limits393EMacroscopicstomach, pylorus- erosion/ulcer, moderatestomach contents red.393EMicroscopicovaries- within normal limitsstomach, glandular- within normal limitsuterus- within normal limitsnon-correlated macro observation- stomach, pylorus - erosion/ulcer394DMacroscopicall tissues- within normal limitsS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 227 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Animal Data Record: Pathology - FEMALETerminalGroup,Animal Number FateTissueObservations2400 mg/kg/dayDMicroscopic394ovaries- within normal limitsuterus- within normal limits395rMacroscopicDall tissues- within normal limits395DMicroscopicovaries- within normal limitsuterus- within normal limitsMacroscopic396Sanimal/whole body- body fat depleted, mildcorresponds to antemortem observation (thin)396SMicroscopicovaries- within normal limitsuterus- within normal limitsnon-correlated macro observation- animal/whole body - body fat depletedS - Scheduled Necropsy E - Euthanizedin extremisD - Died on StudyrReplacement animal 

# Page 228 of 358

# 1271-001 Appendix L

Individual P Premating/Ma ting Estrous Cycling Data Page 229 of 358

# -



# 1271-001 PROCEDURE FOR DETERMINING ESTROUS CYCLE LENGTH

IN RODENTS

The cycle length count (in days) was started from the first day estrous smears were collected.

The starting point for each animals’ cycle was referenced to the first occurrence of estrus. It

was determined how many days there were from the start of the cycle to the first occurrence

of estrus. This number was equal to the number of days prior to estrus and was used as the

cycle starting point for every cycle. More than 3 days prior to the first incidence of estrus

was not counted. If there were more than 3 days prior to the first incidence of estrus, the

count began from the first incidence of estrus.

In situations where estrus occurred on the first day of the cycle (starting point), the number of

days from the start of cycle to the first occurrence was zero (0). In this case, the cycle started

on estrus. If the first occurrence of estrus had 2 or more consecutive days of estrus, the last

estrus was assigned as the start of the first cycle and continued to the last estrus of the next

cycle.

To determine the number of days in a cycle, the number of days it took to go from the

starting point of the first cycle to the starti ng point of the next cycle was counted. The

starting point was not counted as 1 day.

Due to the potential occurrence of prolonged estrus (estrus occurring on 2 or more

consecutive days) or highly irregular cyclicity, the “subsequent” estrus was defined as the

nearest estrus that was preceded by at least 2 days of non-estrus. To count the number of

days in subsequent cycles, the process was started over from where it left off (end of cycle =

start of new cycle). Page 230 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS #84-65-1) in RatsIndividual P Premating Estrous CyclingGroup, Cycle Length Mean Cycle Number ofAnimal (Days) Length CyclesNumber Cycle 1 Cycle 2 Cycle 3 Cycle 4 (Days) (Count)NA - Not Applicable/Not Availablee - Excluded from summary statisticsrReplacement animal0 mg/kg/day3494 4 4 4 4.00 43506 4 6 5.33 3351NA NA NA3525 4 5 3 4.25 43533 4 5 5 4.25 43545 5 5 5.00 33553 5 5 5 4.50 43565 4 4 4 4.25 43574 4 4 4 4.00 43584 4 4 4.00 33595 5 5 4 4.75 43606 4 34.33 3150 mg/kg/day3614 5 4 4.33 33624 4.00 13635 10e4 4.50 23644 8 4 5.33 33654 4 8 5.33 33664 4 4 4 4.00 43674 4.00 13684 4 4 44.00 4369r88.00 13704 4 4 44.00 4371r44.00 13724 6 55.00 3 

# Page 231 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS #84-65-1) in RatsIndividual P Premating Estrous CyclingGroup, Cycle Length Mean Cycle Number ofAnimal (Days) Length CyclesNumber Cycle 1 Cycle 2 Cycle 3 Cycle 4 (Days) (Count)NA - Not Applicable/Not Availablee - Excluded from summary statisticsrReplacement animal600 mg/kg/day373r88.00 1374NANA NA3759 57.00 23767 4 45.00 337712e44.00 1378r4 4 64.67 33793 4 95.33 33809 57.00 2381r4 5 44.33 33828 56.50 2383NANA NA3844 44.00 22400 mg/kg/day385NANA NA3865 4 44.33 33873 4 54.00 3388NANA NA3897 5 45.33 33904 5 85.67 3391r4 4 44.00 3392r5 44.50 23935 5 44.67 33945 5 86.00 3395r55.00 13964 5 54.67 3 

# Page 232 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS #84-65-1) in RatsIndividual P Premating/Mating Estrous Cycle StagesGroup,Animal Day of StudyNumber1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17DI - Diestrus MET - Metestrus PRO - Proestrus EST - EstrusS - SpermrReplacement animal0 mg/kg/day349 DI DI DI EST DI DI DI ESTDI DI DI EST DI DI DI EST DI350 DI EST PRO DI DI DI EST ESTDI DI DI EST MET DI DI DI DI351 PRO DI DI DI DI DI DI DIDI DI DI DI PRO DI DI DI EST352 EST DI DI EST EST EST PRO DIDI EST DI DI DI MET EST DI DI353 EST DI PRO EST DI DI DI ESTDI DI DI EST EST DI DI DI EST354 DI DI DI EST EST DI DI DIDI EST DI DI DI EST EST DI DI355 DI EST PRO EST EST DI DI DIEST EST DI DI DI EST EST DI DI356 DI EST DI DI DI DI EST DIDI DI EST DI DI DI EST DI DI357 DI EST DI DI DI EST DI DIDI EST DI DI DI EST DI DI DI358 DI DI EST DI DI DI EST DIDI DI EST DI DI DI EST DI DI359 EST DI EST DI EST EST PRO DI DI EST EST DI DI DI EST EST DI360 DI DI DI DI EST DI DI DIDI DI EST DI DI DI EST DI DI150 mg/kg/day361 DI DI DI DI DI EST PRO DIDI EST DI DI DI EST EST DI DI362 DI DI PRO PRO DI DI DI DIDI DI DI DI DI PRO DI EST DI363 EST DI DI DI DI EST DI DIDI DI DI DI DI DI DI EST DI364 EST DI DI DI EST DI DI DIPRO DI DI DI EST PRO DI DI EST365 DI PRO DI EST DI DI DI ESTDI DI DI EST DI DI DI DI DI366 DI PRO DI EST DI DI DI ESTDI DI DI EST DI DI DI EST DI367 DI DI DI DI MET DI DI DIDI DI DI DI DI DI EST DI DI368 DI EST DI DI DI EST PRO DIDI EST DI DI DI EST DI DI DI369rDI PRO EST EST DI DI DI DIDI DI DI EST DI DI DI DI S370 DI EST PRO DI DI EST PRO DIDI EST DI DI DI EST DI DI DI371rDI DI DI DI DI DI DI ESTPRO DI DI EST DI DI DI DI S372 DI DI DI EST EST DI DI DIEST DI DI DI DI EST EST DI DI 

# Page 233 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS #84-65-1) in RatsIndividual P Premating/Mating Estrous Cycle StagesGroup,Animal Day of StudyNumber1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17DI - Diestrus PRO - Proestrus EST - Estrus NA - Not Applicable/Not AvailableS - SpermrReplacement animal600 mg/kg/day373rDI EST DI DI DI DI DI DIPRO EST DI DI DI DI DI S374 DI DI DI DI DIDI DI DI DI DI DIDI DI DI DI DI DI375 DI PRO EST EST DI DI DI DIDI DI DI EST EST DI DI DI EST376 DI PRO EST DI DI DI DI DIDI EST DI DI DI EST DI DI DI377 EST EST PRO EST DI DI DI DIDI DI DI DI EST EST DI DI DI378rEST DI DI DI EST DI DI DI ESTDI DI DI DI EST EST DI DI379 DI EST PRO EST EST DI DI DIEST DI DI DI DI DI DI DI PRO380 DI DI EST EST DI DI DI DIDI DI DI EST EST DI DI DI EST381rDI EST DI DI DI EST DI DI DIDI EST DI DI DI EST DI DI382 DI DI DI DI EST DI EST DIDI DI DI DI EST DI DI DI EST383 DI DI DI DI DIDI DI DI DI DI DIDI DI DI DI DI DI384 DI DI DI EST EST DI DI DIEST DI DI DI EST DI DI DI DI2400 mg/kg/day385 DI DI EST EST DI DI DI DIDI DI DI DI DI PRO DI DI DI386 DI DI DI EST EST PRO DI DIDI EST DI DI DI EST DI DI DI387 EST DI DI EST DI DI EST ESTDI DI DI EST EST DI DI DI DI388 DI DI DI DI DIDI DI DI DI DI DIDI DI DI DI DI DI389 EST PRO EST DI DI DI PRO ESTDI DI DI DI EST DI DI DI EST390 DI DI EST DI DI DI EST DIDI DI DI EST DI DI DI DI DI391rEST EST PRO DI DI EST DI DI DIEST DI DI DI EST DI DI DI392rDI NA PRO EST DI DI DI DI ESTPRO DI DI EST DI DI DI EST393 DI DI EST EST DI PRO DI PROEST DI DI DI EST EST DI DI DI394 EST EST DI DI DI DI EST DIDI DI DI EST DI DI DI DI DI395rDI NA DI DI DI DI EST DIDI DI DI EST DI DI DI DI S396 DI DI PRO EST EST DI DI DIEST DI DI DI DI EST DI DI DI 

# Page 234 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS #84-65-1) in RatsIndividual P Premating/Mating Estrous Cycle StagesGroup,Animal Day of StudyNumber18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34DI - Diestrus MET - Metestrus PRO - Proestrus EST - EstrusS - Sperm P - PlugrReplacement animal0 mg/kg/day349 DI DI EST DI PRO DI EST S350 EST DI DI PRO S351 DI DI DI DI S352 EST PRO EST DI DI DI DI DIDI DI DI DI DI DI DI DI DI353 EST DI DI DI EST S354 DI DI EST DI PRO EST S355 DI EST EST DI DI PRO S356 DI EST DI DI PRO EST S357 EST DI DI EST S358 DI PRO DI DI DI S359 DI PRO EST S360 EST MET DI DI EST S150 mg/kg/day361 DI EST DI DI PRO S362 DI PRO EST S363 DI DI EST DI DI DI ESTPRO DI DI DI S364 DI DI DI S365 DI DI EST DI DI DI S366 DI DI EST DI DI DI S367 DI EST DI DI EST EST S368 EST DI DI EST S369r370 EST DI DI DI P371r372 DI EST EST DI DI DI DI DIDI DI DI DI DI DI DI DI DI 

# Page 235 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS #84-65-1) in RatsIndividual P Premating/Mating Estrous Cycle StagesGroup,Animal Day of StudyNumber18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34DI - Diestrus MET - Metestrus PRO - Proestrus EST - EstrusS - Sperm P - PlugrReplacement animal600 mg/kg/day373r374 DI EST DI DI DI S375 EST DI DI DI S376 EST DI DI DI S377 EST DI DI DI EST EST S378rDI S379 EST DI DI DI DI DI EST DIDI DI DI DI DI DI DI DI DI380 EST DI DI DI DI DI DI DIDI DI DI DI DI DI DI DI DI381rDI EST DI DI EST S382 EST DI DI EST S383 DI DI EST DI DI DI S384 DI DI DI DI S2400 mg/kg/day385 DI DI DI DI DIDI DI DI DI DI DIDI DI DI DI DI DI386 EST DI DI DI P387 PRO DI DI EST S388 DI DI DI DI PRO S389 MET DI DI PRO S390 DI EST EST DI DI DI S391rS392rS393 EST DI DI EST P394 DI DI EST DI DI DI DI DIDI DI DI DI DI DI DI DI DI395r396 DI EST DI DI DI EST S 

# Page 236 of 358

# 1271-001 Appendix M

Individual P Mating Record Page 237 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CA # 84-65-1) in RatsIndividual Mating RecordGroup,FemaleNumber Male Number Outcome Dateof Cohabitation Date of InseminationNRMP - Normal pregnancy NA - Not Applicable/Not AvailableNP - Not pregnantrReplacement animal0 mg/kg/day349 303 NRMP 17JUL2006 22JUL2006350 312 NRMP 17JUL2006 19JUL2006351 301 NRMP 17JUL2006 19JUL2006352 305 NP 17JUL2006 NA353 310 NRMP 17JUL2006 20JUL2006354 304 NRMP 17JUL2006 21JUL2006355 307 NRMP 17JUL2006 21JUL2006356 309 NRMP 17JUL2006 21JUL2006357 308 NRMP 17JUL2006 19JUL2006358 306 NRMP 17JUL2006 20JUL2006359 311 NRMP 17JUL2006 18JUL2006360 302 NRMP 17JUL2006 20JUL2006150 mg/kg/day361 320 NRMP 17JUL2006 20JUL2006362 315 NRMP 17JUL2006 18JUL2006363 322 NRMP 17JUL2006 26JUL2006364 314 NRMP 17JUL2006 18JUL2006365 318 NRMP 17JUL2006 21JUL2006366 319 NRMP 17JUL2006 21JUL2006367 317NRMP17JUL2006 21JUL2006368 313NRMP17JUL2006 19JUL2006369r321NRMP17JUL2006 18JUL2006370 316NRMP17JUL2006 19JUL2006371r323 NP 17JUL2006 19JUL2006372 324rNP 17JUL2006 NA 

# Page 238 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CA # 84-65-1) in RatsIndividual Mating RecordGroup,FemaleNumber Male Number Outcome Dateof Cohabitation Date of InseminationALR - All resorptions NRMP - Normal pregnancy NA - Not Applicable/Not AvailableDNP - Died not pregnant PIDE -Pregnant incomplete deliveryrReplacement animalDP - Died pregnant PEM - Pregnant euthanized moribund600 mg/kg/day373r326 NRMP 17JUL2006 18JUL2006374 325 PIDE 17JUL2006 20JUL2006375 328 NRMP 17JUL2006 19JUL2006376 334 PIDE 17JUL2006 19JUL2006377 336 NRMP 17JUL2006 21JUL2006378r331 DP 17JUL2006 21JUL2006379 330 DP 17JUL2006 NA380 335PNMD17JUL2006 NA381r333rNRMP17JUL2006 23JUL2006382 327NRMP17JUL2006 19JUL2006383 332NRMP17JUL2006 21JUL2006384 329NRMP17JUL2006 19JUL20062400 mg/kg/day385 348 DNP 17JUL2006 NA386 338 DP 17JUL2006 19JUL2006387 347 NRMP 17JUL2006 19JUL2006388 346 DP 17JUL2006 20JUL2006389 344 NRMP 17JUL2006 19JUL2006390 341 NRMP 17JUL2006 21JUL2006391r342 DP 17JUL2006 19JUL2006392r343 NRMP 17JUL2006 19JUL2006393 339 PEM 17JUL2006 19JUL2006394 340 DP 17JUL2006 NA395r345 DP 17JUL2006 18JUL2006396 337rALR 17JUL2006 21JUL2006 

# Page 239 of 358

# 1271-001 Appendix N

Individual P Natural Delivery and Litter Data Page 240 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Natural Delivery and Litter Data_________________________________________________________________________________________________________________________________rReplacement animalGroup,FemaleGestationLengthNumber of Pups Delivered on Day 0ImplantPost-ImplantationNumber(Days)LivebornStillbornUncertainTotalScarsLoss (%)0 mg/kg/day349221300131618.835022160016160.035122170017185.635322160016160.035422160016160.035523130013147.135622170017170.0357221300131513.3358221200121414.335922120012137.736022160016160.0150 mg/kg/day36123130013147.136222100010119.136322121013147.1364231300131513.336523140014156.7366231200121414.336723140014140.0368221000101323.1369r23150015150.037022130013147.1 

# Page 241 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Natural Delivery and Litter Data_________________________________________________________________________________________________________________________________PIDE - Pregnant incomplete deliveryrReplacement animalx - Excluded from summary statisticsPNMD - Pregnant no mating dateNA - Not Applicable/Not AvailableGroup,FemaleGestationLengthNumber of Pups Delivered on Day 0ImplantPost-ImplantationNumber(Days)LivebornStillbornUncertainTotalScarsLoss (%)600 mg/kg/day373r2410011190.9374xPIDE233003NANA37524650111315.4376xPIDE23014014140.037723110011110.0380PNMDNA160016175.9381r236006825.0382230130131513.338322122014140.0384231300131513.32400 mg/kg/day3872312031070.038923120012137.7390221300131513.3392r2453081233.3 

# Page 242 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Litter SizeGroup, Number of Live PupsFemale Day 0 Day 4Number M F M FrReplacement animal0 mg/kg/day3494 9 4 93507 9 7 935112 5 12 53539 7 8 53548 8 7 83558 5 8 535611 6 11 53574 9 4 93587 5 7 53595 7 5 736014 2 14 2150 mg/kg/day3616 7 0 03623 7 3 73635 7 5 736410 3 10 33654 10 4 103665 7 5 73677 7 7 73685 5 5 4369r7 8 7 83705 8 5 8 

# Page 243 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual P Litter SizeGroup, Number of Live PupsFemale Day 0 Day 4Number M F M FPIDE - Pregnant incomplete deliveryrReplacement animal x - Excluded from summary statisticsPNMD - Pregnant no mating date600 mg/kg/day373r0 0 0 0374xPIDE 1 2 0 03750 0 0 0376xPIDE 0 0 0 03777 3 0 0380PNMD 11 5 0 0381r0 6 0 03820 0 0 03834 8 2 73846 7 0 12400 mg/kg/day3870 1 0 03893 9 3 939010 3 0 0392r0 0 0 0 

# Page 244 of 358

# 1271-001 Appendix O

Individual F 1 

# Pup Clinical Findings Page 245 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day34901No Abnormalities Detected 0, 434902No Abnormalities Detected 0, 434903No Abnormalities Detected 0, 434904No Abnormalities Detected 0, 434905No Abnormalities Detected 0, 434906No Abnormalities Detected 0, 434907No Abnormalities Detected 0, 434908No Abnormalities Detected 0, 434909No Abnormalities Detected 0, 434910No Abnormalities Detected 0, 434911No Abnormalities Detected 0, 434912No Abnormalities Detected 0, 434913No Abnormalities Detected 0, 435001No Abnormalities Detected 0, 4 

# Page 246 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35002No Abnormalities Detected 0, 435003No Abnormalities Detected 0, 435004No Abnormalities Detected 0, 435005No Abnormalities Detected 0, 435006No Abnormalities Detected 0, 435007No Abnormalities Detected 0, 435008No Abnormalities Detected 0, 435009No Abnormalities Detected 0, 435010No Abnormalities Detected 0, 435011No Abnormalities Detected 0, 435012No Abnormalities Detected 0, 435013No Abnormalities Detected 0, 435014No Abnormalities Detected 0, 435015No Abnormalities Detected 0, 4 

# Page 247 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35016No Abnormalities Detected 0, 435101No Abnormalities Detected 0, 435102No Abnormalities Detected 0, 435103No Abnormalities Detected 0, 435104No Abnormalities Detected 0, 435105No Abnormalities Detected 0, 435106No Abnormalities Detected 0, 435107No Abnormalities Detected 0, 435108No Abnormalities Detected 0, 435109No Abnormalities Detected 0, 435110No Abnormalities Detected 0, 435111No Abnormalities Detected 0, 435112No Abnormalities Detected 0, 435113No Abnormalities Detected 0, 4 

# Page 248 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35114No Abnormalities Detected 0, 435115No Abnormalities Detected 0, 435116No Abnormalities Detected 0, 435117No Abnormalities Detected 0, 435301No Abnormalities Detected 0, 435302No Abnormalities Detected 0, 435303No Abnormalities Detected 0, 435304No Abnormalities Detected 0, 435305No Abnormalities Detected 0, 435306No Abnormalities Detected 0, 435307No Abnormalities Detected 0, 435308No Abnormalities Detected 0, 435309No Abnormalities Detected 035310No Abnormalities Detected 0 

# Page 249 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35311No Abnormalities Detected 035312No Abnormalities Detected 0, 435313No Abnormalities Detected 0, 435314No Abnormalities Detected 0, 435315No Abnormalities Detected 0, 435316No Abnormalities Detected 0, 435401No Abnormalities Detected 0, 435402No Abnormalities Detected 0, 435403No Abnormalities Detected 0, 435404No Abnormalities Detected 0, 435405No Abnormalities Detected 0, 435406No Abnormalities Detected 0, 435407No Abnormalities Detected 0, 435409No Abnormalities Detected 0, 4 

# Page 250 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35410No Abnormalities Detected 0, 435411No Abnormalities Detected 0, 435412No Abnormalities Detected 0, 435413No Abnormalities Detected 0, 435414No Abnormalities Detected 0, 435415No Abnormalities Detected 0, 435416No Abnormalities Detected 0, 435501No Abnormalities Detected 0, 435502No Abnormalities Detected 0, 435503No Abnormalities Detected 0, 435504No Abnormalities Detected 0, 435505No Abnormalities Detected 0, 435506No Abnormalities Detected 0, 435507No Abnormalities Detected 0, 4 

# Page 251 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35508No Abnormalities Detected 0, 435509No Abnormalities Detected 0, 435510No Abnormalities Detected 0, 435511No Abnormalities Detected 0, 435512No Abnormalities Detected 0, 435513No Abnormalities Detected 0, 435601No Abnormalities Detected 0, 435602No Abnormalities Detected 0, 435603No Abnormalities Detected 0, 435604No Abnormalities Detected 0, 435605No Abnormalities Detected 0, 435606No Abnormalities Detected 0, 435607No Abnormalities Detected 0, 435608No Abnormalities Detected 0, 4 

# Page 252 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35609No Abnormalities Detected 0, 435610No Abnormalities Detected 0, 435611No Abnormalities Detected 0, 435612No Abnormalities Detected 0, 435613No Abnormalities Detected 0, 435614No Abnormalities Detected 0, 435615No Abnormalities Detected 035616No Abnormalities Detected 0, 435617No Abnormalities Detected 0, 435701No Abnormalities Detected 0, 435702No Abnormalities Detected 0, 435703No Abnormalities Detected 0, 435704No Abnormalities Detected 0, 435705No Abnormalities Detected 0, 4 

# Page 253 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35706No Abnormalities Detected 0, 435707No Abnormalities Detected 0, 435708No Abnormalities Detected 0, 435709No Abnormalities Detected 0, 435710No Abnormalities Detected 0, 435711No Abnormalities Detected 0, 435712No Abnormalities Detected 0, 435713No Abnormalities Detected 0, 435801No Abnormalities Detected 0, 435802No Abnormalities Detected 0, 435803No Abnormalities Detected 0, 435804No Abnormalities Detected 0, 435805No Abnormalities Detected 0, 435806No Abnormalities Detected 0, 4 

# Page 254 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35807No Abnormalities Detected 0, 435808No Abnormalities Detected 0, 435809No Abnormalities Detected 0, 435810No Abnormalities Detected 0, 435811No Abnormalities Detected 0, 435812No Abnormalities Detected 0, 435901No Abnormalities Detected 0, 435902No Abnormalities Detected 0, 435903No Abnormalities Detected 0, 435904No Abnormalities Detected 0, 435905No Abnormalities Detected 0, 435906No Abnormalities Detected 0, 435907No Abnormalities Detected 0, 435908No Abnormalities Detected 0, 4 

# Page 255 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day35909No Abnormalities Detected 0, 435910No Abnormalities Detected 0, 435911No Abnormalities Detected 0, 435912No Abnormalities Detected 0, 436001No Abnormalities Detected 0, 436002No Abnormalities Detected 0, 436003No Abnormalities Detected 0, 436004No Abnormalities Detected 0, 436005No Abnormalities Detected 0, 436006No Abnormalities Detected 0, 436007No Abnormalities Detected 0, 436008No Abnormalities Detected 0, 436009No Abnormalities Detected 0, 436010No Abnormalities Detected 0, 4 

# Page 256 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present0 mg/kg/day36011No Abnormalities Detected 0, 436012No Abnormalities Detected 0, 436013No Abnormalities Detected 0, 436014No Abnormalities Detected 0, 436015No Abnormalities Detected 0, 436016No Abnormalities Detected 0, 4 

# Page 257 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day36101No Abnormalities Detected 036102No Abnormalities Detected 036103No Abnormalities Detected 036104No Abnormalities Detected 036105No Abnormalities Detected 036106No Abnormalities Detected 036107No Abnormalities Detected 036108No Abnormalities Detected 036109No Abnormalities Detected 036110No Abnormalities Detected 036111No Abnormalities Detected 036112No Abnormalities Detected 036113No Abnormalities Detected 036201No Abnormalities Detected 0, 4 

# Page 258 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day36202No Abnormalities Detected 0, 436203No Abnormalities Detected 0, 436204No Abnormalities Detected 0Abrasion(s), Cranial region 4Skin cold to touch 4Skin discolored, Pale, Entire body 436205No Abnormalities Detected 0, 436206No Abnormalities Detected 0, 436207No Abnormalities Detected 0, 436208No Abnormalities Detected 0, 436209No Abnormalities Detected 0, 436210No Abnormalities Detected 0, 436301No Abnormalities Detected 0, 436302No Abnormalities Detected 0, 436303No Abnormalities Detected 0, 4 

# Page 259 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day36304No Abnormalities Detected 0, 436305No Abnormalities Detected 0, 436306No Abnormalities Detected 0, 436307No Abnormalities Detected 0, 436308No Abnormalities Detected 0, 436309No Abnormalities Detected 0, 436310No Abnormalities Detected 0, 436311No Abnormalities Detected 0, 436312No Abnormalities Detected 0, 436401No Abnormalities Detected 0, 436402No Abnormalities Detected 0, 436403No Abnormalities Detected 0, 436404No Abnormalities Detected 0, 436405No Abnormalities Detected 0, 4 

# Page 260 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day36406No Abnormalities Detected 0, 436407No Abnormalities Detected 0, 436408No Abnormalities Detected 0, 436409No Abnormalities Detected 0, 436410No Abnormalities Detected 0, 436411No Abnormalities Detected 0, 436412No Abnormalities Detected 0, 436413No Abnormalities Detected 0, 436501No Abnormalities Detected 0, 436502No Abnormalities Detected 0, 436503No Abnormalities Detected 0, 436504No Abnormalities Detected 0, 436505No Abnormalities Detected 0, 436506No Abnormalities Detected 0, 4 

# Page 261 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day36507No Abnormalities Detected 0, 436508No Abnormalities Detected 0, 436509No Abnormalities Detected 0, 436510No Abnormalities Detected 0, 436511No Abnormalities Detected 0, 436512No Abnormalities Detected 0, 436513No Abnormalities Detected 0, 436514No Abnormalities Detected 0, 436601No Abnormalities Detected 0, 436602No Abnormalities Detected 0, 436603No Abnormalities Detected 0, 436604No Abnormalities Detected 0, 436605No Abnormalities Detected 0, 436606No Abnormalities Detected 0, 4 

# Page 262 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day36607No Abnormalities Detected 0, 436608No Abnormalities Detected 0, 436609No Abnormalities Detected 0, 436610No Abnormalities Detected 0, 436611No Abnormalities Detected 0, 436612No Abnormalities Detected 0, 436701No Abnormalities Detected 0, 436702No Abnormalities Detected 0, 436703No Abnormalities Detected 0, 436704No Abnormalities Detected 0, 436705No Abnormalities Detected 0, 436706No Abnormalities Detected 0, 436707No Abnormalities Detected 0, 436708No Abnormalities Detected 0, 4 

# Page 263 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day36709No Abnormalities Detected 0, 436710No Abnormalities Detected 0, 436711No Abnormalities Detected 0, 436712No Abnormalities Detected 0, 436713No Abnormalities Detected 0, 436714No Abnormalities Detected 0, 436801No Abnormalities Detected 0, 436802No Abnormalities Detected 0, 436803No Abnormalities Detected 0, 436804No Abnormalities Detected 0, 436805No Abnormalities Detected 4Limb function impaired, Hind limb/left 0Limb function impaired, Hind limb/right 036806No Abnormalities Detected 0, 436807No Abnormalities Detected 0, 4 

# Page 264 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign PresentrReplacement animal150 mg/kg/day36808No Abnormalities Detected 036809No Abnormalities Detected 4Limb function impaired, Hind limb/left 0Limb function impaired, Hind limb/right 036810No Abnormalities Detected 0, 436901rNo Abnormalities Detected 0, 436902rNo Abnormalities Detected 0, 436903rNo Abnormalities Detected 0, 436904rNo Abnormalities Detected 0, 436905rNo Abnormalities Detected 0, 436906rNo Abnormalities Detected 0, 436907rNo Abnormalities Detected 0, 436908rNo Abnormalities Detected 0, 436909rNo Abnormalities Detected 0, 436910rNo Abnormalities Detected 0, 4 

# Page 265 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign PresentrReplacement animal150 mg/kg/day36911rNo Abnormalities Detected 0, 436912rNo Abnormalities Detected 0, 436913rNo Abnormalities Detected 0, 436914rNo Abnormalities Detected 0, 436915rNo Abnormalities Detected 0, 437001No Abnormalities Detected 0, 437002No Abnormalities Detected 0, 437003No Abnormalities Detected 0, 437004No Abnormalities Detected 0, 437005No Abnormalities Detected 0, 437006No Abnormalities Detected 0, 437007No Abnormalities Detected 0, 437008No Abnormalities Detected 0, 437009No Abnormalities Detected 0, 4 

# Page 266 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present150 mg/kg/day37010No Abnormalities Detected 0, 437011No Abnormalities Detected 0, 437012No Abnormalities Detected 0, 437013No Abnormalities Detected 0, 4 

# Page 267 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present600 mg/kg/day37401Skin cold to touch 037402No Abnormalities Detected 037701Skin cold to touch 037702Skin cold to touch 037703Skin cold to touch 037704Skin cold to touch 037705Skin cold to touch 037706Skin cold to touch 037707Skin cold to touch 037708Skin cold to touch 037709Skin cold to touch 037710Skin cold to touch 038001Skin cold to touch 038002Skin cold to touch 0 

# Page 268 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present600 mg/kg/day38003Activity decreased 0Skin cold to touch 0Skin discolored, Pale, Entire body 038004Skin cold to touch 038005Skin cold to touch 038006Skin cold to touch 038007Skin cold to touch 038008Skin cold to touch 038009Skin cold to touch 038010Skin cold to touch 038011Skin cold to touch 038012Skin cold to touch 038013Skin cold to touch 038014Skin cold to touch 038015Skin cold to touch 0 

# Page 269 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign PresentrReplacement animal600 mg/kg/day38016Skin cold to touch 038101rSkin cold to touch 038102rSkin cold to touch 038103rSkin cold to touch 038104rSkin cold to touch 038105rSkin cold to touch 038106rSkin cold to touch 038301No Abnormalities Detected 0, 438302No Abnormalities Detected 0, 438303No Abnormalities Detected 038304No Abnormalities Detected 038305No Abnormalities Detected 0, 438306No Abnormalities Detected 0, 438307No Abnormalities Detected 0, 4 

# Page 270 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present600 mg/kg/day38308No Abnormalities Detected 038309No Abnormalities Detected 0, 438310No Abnormalities Detected 0, 438311No Abnormalities Detected 0, 438312No Abnormalities Detected 0, 438313No Abnormalities Detected 038401No Abnormalities Detected 038402No Abnormalities Detected 038403No Abnormalities Detected 038404No Abnormalities Detected 038405No Abnormalities Detected 038407No Abnormalities Detected 038408No Abnormalities Detected 038409No Abnormalities Detected 0 

# Page 271 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present600 mg/kg/day38410No Abnormalities Detected 038411No Abnormalities Detected 038412No Abnormalities Detected 0Thin 438413No Abnormalities Detected 0 

# Page 272 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present2400 mg/kg/day38901No Abnormalities Detected 0, 438902No Abnormalities Detected 0, 438903No Abnormalities Detected 0, 438904No Abnormalities Detected 0, 438905No Abnormalities Detected 0, 438906No Abnormalities Detected 0, 438907No Abnormalities Detected 0, 438908No Abnormalities Detected 0, 438909No Abnormalities Detected 0, 438910No Abnormalities Detected 0, 438911No Abnormalities Detected 0, 438912No Abnormalities Detected 0, 439001No Abnormalities Detected 039002No Abnormalities Detected 0 

# Page 273 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1PupClinical FindingsGroup,Animal Number Lactation Day Sign Present2400 mg/kg/day39003No Abnormalities Detected 039004No Abnormalities Detected 039005No Abnormalities Detected 039006No Abnormalities Detected 039007No Abnormalities Detected 039008No Abnormalities Detected 039009No Abnormalities Detected 039010No Abnormalities Detected 039011No Abnormalities Detected 039012No Abnormalities Detected 039013No Abnormalities Detected 0 

# Page 274 of 358

# 1271-001 Appendix P

Individual F 1 

# Pup Body Weight, Sex, and Status Data Page 275 of 358

# 1271-001 Code Order For Individual Pup Sex and Status

The following table presents the sex and fate of each pup on study.

The first letter represents the sex of the pup. The second letter represents the fate of the pup.

The number represents the postnatal day on which the fate occurred.

For example, MD3 represents the following information:

M – Male

D – Died

3 – Postnatal Day 3 (day of fate)

Each letter/number combination corresponds to the dam to which the pup was born (at left) and

the identification number to which it was as signed (ascending numbers across the top).

The dam number and the two-digit (e.g., 01) identification number comprise the pup number. 

# Page 276 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Body Weight Values, gGroup,Postpartum Day 0FemalePup #Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17D - DiedrReplacement animal0 mg/kg/day349 7.2 6.8 7.2 6.8 6.3 6.3 6.9 7.0 7.0 6.4 6.4 7.2 6.3350 6.3 7.0 6.6 7.26.3 6.7 6.4 6.9 6.1 6.8 6.4 6.4 6.4 6.1 5.7 6.1351 8.0 7.6 6.5 7.38.2 6.2 6.9 7.0 6.4 7.0 6.8 7.4 7.36.5 6.7 6.2 6.5353 6.2 6.4 6.8 5.56.2 6.2 6.6 5.7 6.1 5.8 5.5 5.6 6.7 7.0 6.3 5.5354 6.2 6.3 5.3 6.05.9 6.4 6.0 D 5.8 5.9 5.7 5.6 5.6 5.5 5.4 5.4355 7.4 7.8 7.2 7.3 7.0 7.6 8.2 7.3 6.9 7.2 7.2 7.0 7.3356 6.8 6.3 6.8 6.56.2 5.7 5.8 6.7 6.4 5.0 5.3 6.6 6.96.2 6.4 6.7 6.5357 7.0 7.2 6.6 7.3 5.4 6.6 6.5 6.7 6.4 6.7 5.9 8.8 6.5358 7.1 7.0 6.8 7.1 7.2 6.6 5.8 6.2 6.96.7 6.5 6.1359 6.5 6.3 6.6 7.0 7.3 6.0 6.2 6.7 5.56.3 6.1 6.1360 7.8 7.4 6.7 7.87.2 7.0 6.9 6.8 7.1 7.1 6.4 7.3 7.0 6.9 6.7 6.8150 mg/kg/day361 6.1 6.2 6.5 5.9 4.9 5.4 5.6 6.4 5.7 5.4 5.0 4.7 4.3362 6.8 6.5 6.0 6.8 6.7 6.1 6.6 7.4 6.3 6.5363 6.1 6.1 6.1 5.3 5.8 5.9 5.0 6.1 5.25.9 5.8 5.8 S364 6.3 6.4 6.2 7.4 6.2 6.5 5.9 6.1 5.8 6.1 6.1 5.5 6.3365 7.7 7.6 7.0 7.6 7.3 6.5 7.3 7.4 7.7 7.0 6.7 6.5 7.3 7.0366 6.5 6.4 5.6 6.5 6.7 6.3 6.2 6.2 6.46.6 5.8 6.2367 6.3 7.8 7.4 7.2 7.2 7.3 7.2 7.5 6.3 6.6 7.2 7.2 6.6 6.7368 6.5 7.6 7.1 7.1 7.0 6.6 6.5 6.8 6.6 6.8369r6.6 6.8 6.2 5.9 6.3 6.1 6.6 5.6 6.0 6.3 6.2 5.7 6.4 6.5 5.7370 7.1 6.6 7.8 7.2 7.0 6.9 6.7 6.9 6.5 6.2 6.9 6.5 7.7 

# Page 277 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Body Weight Values, gGroup,Postpartum Day 0FemalePup #Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17D - DiedrReplacement animal x - Excluded from summary statisticsS - Stillborn PIDE - Pregnant incomplete deliveryY - Cannibalized600 mg/kg/day373rY374 x PIDE 5.4 5.0 D375 S S S S S D Y Y Y Y Y376 x PIDE S S S S S S S S S S S S S S377 4.1 4.2 4.4 4.7 4.7 4.2 4.5 4.4 4.6 4.2 Y380 4.6 4.3 3.7 3.7 3.9 4.1 4.3 4.6 3.7 3.9 3.8 3.6 4.0 4.0 4.0 3.5381r7.4 7.0 7.2 6.6 6.2 6.7382 S S S S S S S S S S S S S383 5.8 5.8 5.4 5.15.5 4.0 5.5 4.8 5.0 5.2 4.8 5.6 4.8 S384 6.0 5.9 5.9 5.3 6.4 D 5.5 5.0 4.7 5.3 5.8 5.1 3.92400 mg/kg/day387 S S D389 6.6 6.8 6.4 6.3 6.1 6.0 5.9 5.7 6.26.5 6.2 5.6390 6.7 6.0 6.5 6.4 6.7 8.4 6.7 6.1 6.1 6.4 5.4 5.7 6.6392rS S S D Y Y Y Y 

# Page 278 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Body Weight Values, gGroup,Postpartum Day 4FemalePup #Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17D - DiedrReplacement animalZ - Missing from cage0 mg/kg/day349 10.7 10.7 10.1 9.6 9.7 9.2 9.8 9.6 10.5 9.7 10.2 10.2 9.6350 9.5 10.2 9.9 9.48.2 9.4 9.8 9.4 8.5 8.9 9.7 9.1 8.7 8.6 9.1 8.9351 12.8 10.7 9.6 10.611.1 9.3 11.3 10.3 9.5 10.3 10.9 11.3 11.410.7 10.1 10.1 10.9353 8.8 8.9 8.4 6.3 7.9 8.2 9.0 7.6 D Z D6.3 7.6 8.1 7.9 6.5354 8.1 9.2 7.9 7.99.2 9.6 8.9 D 8.9 8.4 7.7 7.9 8.3 7.5 8.1 8.1355 9.6 11.3 10.0 9.9 11.1 9.5 11.4 10.0 10.8 9.3 9.1 9.9 10.7356 10.2 8.8 9.1 9.57.9 8.5 8.4 9.9 8.6 8.2 10.1 9.7 9.5 7.8 D 10.2 9.6357 10.4 9.5 10.4 10.1 7.1 9.0 8.0 10.0 9.0 10.4 7.5 10.2 9.7358 9.6 11.4 10.8 10.3 11.1 10.0 8.8 9.1 9.69.9 9.9 9.1359 10.3 9.4 9.9 9.6 10.3 9.0 9.1 9.6 9.59.6 9.0 10.1360 11.0 10.9 9.5 10.711.0 9.7 9.9 10.0 9.8 9.5 9.0 10.5 10.5 10.6 10.1 10.2150 mg/kg/day361 D D D D D D D D D D D D D362 7.4 7.9 7.3 6.4 7.8 7.6 8.0 7.8 7.9 7.9363 7.7 7.2 7.4 6.8 6.9 7.3 6.6 7.5 6.87.1 6.6 7.3 S364 8.6 8.2 8.1 9.7 9.2 9.2 8.4 7.5 7.9 8.0 8.1 7.5 8.4365 11.9 12.0 10.2 10.5 10.7 10.9 11.3 11.0 11.2 11.2 8.2 10.2 10.0 10.9366 9.4 7.9 8.2 7.3 9.2 8.8 7.3 7.4 8.58.9 7.9 8.4367 9.2 10.0 10.5 10.7 10.3 9.5 10.3 10.0 9.3 9.4 9.7 9.5 9.9 9.4368 8.9 9.6 10.2 9.9 7.7 9.4 9.2 D 8.9 10.0369r9.4 9.0 8.9 8.6 7.7 8.0 8.2 8.6 8.9 9.1 8.2 7.8 8.4 8.4 6.4370 8.0 7.8 8.7 8.7 7.2 7.4 8.0 8.2 8.1 7.8 8.4 7.9 9.4 

# Page 279 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Body Weight Values, gGroup,Postpartum Day 4FemalePup #Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17D - Died Z - Missing from cage x - Excluded from summary statisticsS - StillbornrReplacement animalE - Euthanizedin extremisPIDE - Pregnant incomplete deliveryY - Cannibalized600 mg/kg/day373rY374 X PIDE E E D375S S S S S D Y Y Y Y Y376 X PIDE S S S S S S S S S S S S S S377Z D Z D D Z D D Z D Y380D D D D D D Y D D D D D D D D D381rE D D D E D382S S S S S S S S S S S S S3834.7 4.9 D D 4.4 3.3 4.1S 4.0 4.5 3.9 4.2 D S384D D D D D D D D D D D 3.8 D2400 mg/kg/day387 S S D389 8.8 8.5 8.8 8.7 8.4 7.4 8.8 8.5 8.69.0 8.8 7.1390 D Z D Z Z Z D D Z D D D D392rS S S D Y Y Y Y 

# Page 280 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Sex and StatusGroup,FemalePup #Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17SEX CODES: M - Male F - FemalePUP STATUS CODES: N - Scheduled necropsy D - Died Z - Missing from cagerReplacement animal0 mg/kg/day349 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4350 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4351 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4353 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MD 4 FZ 2 FD 2 FN 4 FN 4 FN 4 FN 4 FN 4354 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MD 0 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4355 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4356 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FD 4 FN 4 FN 4357 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4358 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4359 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4360 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4150 mg/kg/day361 MD 3 MD 1 MD 3 MD 3 MD 1 MD 2 FD 3 FD 3 FD 3 FD 2 FD 2 FD 3 FD 2362 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4363 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FS 0364 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4365 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4366 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4367 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4368 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FD 2 FN 4 FN 4369rMN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4370 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 

# Page 281 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Sex and StatusGroup,FemalePup #Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17SEX CODES: M - MaleF - Female U - UnsexedPUP STATUS CODES: N - Scheduled necropsy Y - CannibalizedS - Stillborn D - Died Z - Missing from cage E - Euthanizedin extremisrReplacement animal1Pregnant incomplete delivery600 mg/kg/day373rUY 03741ME 0 FE 0 FD 0375 MS 0 MS 0 FS 0 FS 0 FS 0 UD 0 UY 0 UY 0 UY 0 UY 0 UY 03761MS 0 MS 0 MS 0 FS 0 FS 0 US 0 US 0 US 0 US 0 US 0 US 0 US 0 US 0 US 0377 MZ 1 MD 1 MZ 1 MD 1 MD 1 MZ 1 MD 1 FD 1 FZ 2 FD 1 UY 0380 MD 1 MD 2 MD 1 MD 1 MD 2 MD 2 MY 2 MD 1 MD 1 MD 2 MD 2 FD 1 FD 1 FD 2 FD 2 FD 2381rFE 3 FD 3 FD 3 FD 3 FE 3 FD 3382 MS 0 MS 0 MS 0 MS 0 MS 0 FS 0 FS 0 FS 0 FS 0 FS 0 FS 0 FS 0 FS 0383 MN 4 MN 4 MD 1 MD 0 FN 4 FN 4 FN 4 FS 0 FN 4 FN 4 FN 4 FN 4 FD 4 FS 0384 MD 3 MD 1 MD 1 MD 3 MD 2 MD 0 FD 3 FD 2 FD 2 FD 4 FD 2 FN 4 FD 32400 mg/kg/day387 MS 0 FS 0 FD 0389 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4390 MD 2 MZ 3 MD 3 MZ 3 MZ 3 MZ 3 MD 2 MD 2 MZ 3 MD 2 FD 3 FD 2 FD 2392rMS 0 MS 0 FS 0 UD 0 UY 0 UY 0 UY 0 UY 0 

# Page 282 of 358

# 1271-001 Appendix Q

Individual F 1 

# Pup Animal Data Record: Pathology Page 283 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyStillborn PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study B – Stillborn U – Unable to Determine Sex NT – Not Tattooed363-NTFBMacroscopicexternal appearance- within normal limits 

# Page 284 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyStillborn PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study B – Stillborn U – Unable to Determine Sex NT – Not Tattooed375-NT1MBMacroscopicexternal appearance- within normal limits375-NT2MBMacroscopicexternal appearance- within normal limits375-NT3FBMacroscopicexternal appearance- within normal limits375-NT4FBMacroscopicexternal appearance- within normal limits375-NT5FBMacroscopicexternal appearance- within normal limits376-NT1MBMacroscopicexternal appearance- within normal limits376-NT2MBMacroscopicexternal appearance- within normal limits376-NT3MBMacroscopicexternal appearance- within normal limits376-NT4FBMacroscopicexternal appearance- within normal limits376-NT5FBMacroscopicexternal appearance- within normal limits 

# Page 285 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyStillborn PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study B – Stillborn U – Unable to Determine Sex NT – Not Tattooed382-NT1MBMacroscopicexternal appearance- within normal limits382-NT2MBMacroscopicexternal appearance- within normal limits382-NT3MBMacroscopicexternal appearance- within normal limits382-NT4MBMacroscopicexternal appearance- within normal limits382-NT5MBMacroscopicexternal appearance- within normal limits382-NT6FBMacroscopicexternal appearance- within normal limits382-NT7FBMacroscopicexternal appearance- within normal limits382-NT8FBMacroscopicexternal appearance- within normal limits382-NT9FBMacroscopicexternal appearance- within normal limits382-NT10FBMacroscopicexternal appearance- within normal limits 

# Page 286 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyStillborn PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study B – Stillborn U – Unable to Determine Sex NT – Not Tattooed382-NT11FBMacroscopicexternal appearance- within normal limits382-NT12FBMacroscopicexternal appearance- within normal limits382-NT13FBMacroscopicexternal appearance- within normal limits383-NT1FBMacroscopicexternal appearance- within normal limits383-NT2FBMacroscopicexternal appearance- within normal limits 

# Page 287 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyStillborn PupsGroup,Animal NumberSexFate TissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study B – Stillborn U – Unable to Determine Sex NT – Not Tattooed387-NT1MBMacroscopicexternal appearance- within normal limits387-NT2FBMacroscopicexternal appearance- within normal limits392-NT1MBMacroscopicexternal appearance- within normal limits392-NT2MBMacroscopicexternal appearance- within normal limits392-NT3FBMacroscopicexternal appearance- within normal limits 

# Page 288 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed353-09MDMacroscopicexternal appearance- within normal limits353-11FDMacroscopicexternal appearance- within normal limits354-NTMDMacroscopicexternal appearance- within normal limits356-15FDMacroscopicexternal appearance- within normal limits 

# Page 289 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed361-01MDMacroscopicexternal appearance- within normal limits361-02MDMacroscopicexternal appearance- within normal limits361-03MDMacroscopicexternal appearance- within normal limits361-04MDMacroscopicexternal appearance- within normal limits361-05MDMacroscopicexternal appearance- within normal limits361-06MDMacroscopicexternal appearance- within normal limits361-07FDMacroscopicexternal appearance- within normal limits361-08FDMacroscopicexternal appearance- within normal limits361-09FDMacroscopicexternal appearance- within normal limits361-10FDMacroscopicexternal appearance- within normal limits 

# Page 290 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed361-11FDMacroscopicexternal appearance- within normal limits361-12FDMacroscopicexternal appearance- within normal limits361-13FDMacroscopicexternal appearance- within normal limits368-08FDMacroscopicexternal appearance- within normal limits 

# Page 291 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed374-01MEMacroscopicexternal appearance- within normal limits374-02FEMacroscopicexternal appearance- within normal limits377-02MDMacroscopicexternal appearance- within normal limits377-04MDMacroscopicexternal appearance- within normal limits377-05MDMacroscopicexternal appearance- within normal limits377-07MDMacroscopicexternal appearance- within normal limits377-08FDMacroscopicexternal appearance- within normal limits377-10FDMacroscopicexternal appearance- within normal limits380-01MDMacroscopicexternal appearance- within normal limits380-02MDMacroscopicexternal appearance- within normal limits 

# Page 292 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed380-03MDMacroscopicexternal appearance- within normal limits380-04MDMacroscopicexternal appearance- within normal limits380-05MDMacroscopicexternal appearance- within normal limits380-06MDMacroscopicexternal appearance- within normal limits380-08MDMacroscopicexternal appearance- within normal limits380-09MDMacroscopicexternal appearance- within normal limits380-10MDMacroscopicexternal appearance- within normal limits380-11MDMacroscopicexternal appearance- within normal limits380-12FDMacroscopicexternal appearance- within normal limits380-13FDMacroscopicexternal appearance- within normal limits 

# Page 293 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed380-14FDMacroscopicexternal appearance- within normal limits380-15FDMacroscopicexternal appearance- within normal limits380-16FDMacroscopicexternal appearance- within normal limits381-01FEMacroscopicexternal appearance- within normal limits381-02FDMacroscopicexternal appearance- within normal limits381-03FDMacroscopicexternal appearance- within normal limits381-04FDMacroscopicexternal appearance- within normal limits381-05FEMacroscopicexternal appearance- within normal limits381-06FDMacroscopicexternal appearance- within normal limits383-03MDMacroscopicexternal appearance- within normal limits 

# Page 294 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed383-04MDMacroscopicexternal appearance- within normal limits383-13FDMacroscopicexternal appearance- within normal limits384-NTMDMacroscopicexternal appearance- within normal limits384-01MDMacroscopicexternal appearance- within normal limits384-02MDMacroscopicexternal appearance- within normal limits384-03MDMacroscopicexternal appearance- within normal limits384-04MDMacroscopicexternal appearance- within normal limits384-05MDMacroscopicexternal appearance- within normal limits384-06FDMacroscopicexternal appearance- within normal limits384-07FDMacroscopicexternal appearance- within normal limits 

# Page 295 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed384-08FDMacroscopicexternal appearance- within normal limits384-09FDMacroscopicexternal appearance- within normal limits384-10FDMacroscopicexternal appearance- within normal limits384-13FDMacroscopicexternal appearance- within normal limits 

# Page 296 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied On Study PupsGroup,Animal NumberSexFate TissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed387-NT3FDMacroscopicexternal appearance- within normal limits390-01MDMacroscopicexternal appearance- within normal limits390-03MDMacroscopicexternal appearance- within normal limits390-07MDMacroscopicexternal appearance- within normal limits390-08MDMacroscopicexternal appearance- within normal limits390-10MDMacroscopicexternal appearance- within normal limits390-11FDMacroscopicexternal appearance- within normal limits390-12FDMacroscopicexternal appearance- within normal limits390-13FDMacroscopicexternal appearance- within normal limits 

# Page 297 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied on Study- Unsexed PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed B - Stillborn373-NTUDMacroscopicexternal appearance- too cannibalized to examine375-NT6UDMacroscopicexternal appearance- too cannibalized to examine375-NT7UDMacroscopicexternal appearance- too cannibalized to examine375-NT8UDMacroscopicexternal appearance- too cannibalized to examine375-NT9UDMacroscopicexternal appearance- too cannibalized to examine375-NT10UDMacroscopicexternal appearance- too cannibalized to examine375-NT11UDMacroscopicexternal appearance- too cannibalized to examine376-NT6UBMacroscopicexternal appearance- within normal limits376-NT7UBMacroscopicexternal appearance- within normal limits376-NT8UBMacroscopicexternal appearance- within normal limits 

# Page 298 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied on Study- Unsexed PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed B - Stillborn376-NT9UBMacroscopicexternal appearance- within normal limits376-NT10UBMacroscopicexternal appearance- within normal limits376-NT11UBMacroscopicexternal appearance- within normal limits376-NT12UBMacroscopicexternal appearance- within normal limits376-NT13UBMacroscopicexternal appearance- within normal limits376-NT14UBMacroscopicexternal appearance- within normal limits377-NTUDMacroscopicexternal appearance- too cannibalized to examine380-07UDMacroscopicexternal appearance- too cannibalized to examine 

# Page 299 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDied on Study- Unsexed PupsGroup,Animal NumberSexFate TissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed B - Stillborn392-NT4UDMacroscopicexternal appearance- too cannibalized to examine392-NT5UDMacroscopicexternal appearance- too cannibalized to examine392-NT6UDMacroscopicexternal appearance- too cannibalized to examine392-NT7UDMacroscopicexternal appearance- too cannibalized to examine392-NT8UDMacroscopicexternal appearance- too cannibalized to examine 

# Page 300 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed349-01MSMacroscopicexternal appearance- within normal limits349-02MSMacroscopicexternal appearance- within normal limits349-03MSMacroscopicexternal appearance- within normal limits349-04FSMacroscopicexternal appearance- within normal limits349-05FSMacroscopicexternal appearance- within normal limits349-06FSMacroscopicexternal appearance- within normal limits349-07MSMacroscopicexternal appearance- within normal limits349-08FSMacroscopicexternal appearance- within normal limits349-09FSMacroscopicexternal appearance- within normal limits349-10FSMacroscopicexternal appearance- within normal limits 

# Page 301 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed349-11FSMacroscopicexternal appearance- within normal limits349-12FSMacroscopicexternal appearance- within normal limits349-13FSMacroscopicexternal appearance- within normal limits350-01MSMacroscopicexternal appearance- within normal limits350-02MSMacroscopicexternal appearance- within normal limits350-03MSMacroscopicexternal appearance- within normal limits350-04MSMacroscopicexternal appearance- within normal limits350-05MSMacroscopicexternal appearance- within normal limits350-06MSMacroscopicexternal appearance- within normal limits350-07MSMacroscopicexternal appearance- within normal limits 

# Page 302 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed350-08FSMacroscopicexternal appearance- within normal limits350-09FSMacroscopicexternal appearance- within normal limits350-10FSMacroscopicexternal appearance- within normal limits350-11FSMacroscopicexternal appearance- within normal limits350-12FSMacroscopicexternal appearance- within normal limits350-13FSMacroscopicexternal appearance- within normal limits350-14FSMacroscopicexternal appearance- within normal limits350-15FSMacroscopicexternal appearance- within normal limits350-16FSMacroscopicexternal appearance- within normal limits351-01MSMacroscopicexternal appearance- within normal limits 

# Page 303 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed351-02MSMacroscopicexternal appearance- within normal limits351-03MSMacroscopicexternal appearance- within normal limits351-04MSMacroscopicexternal appearance- within normal limits351-05MSMacroscopicexternal appearance- within normal limits351-06MSMacroscopicexternal appearance- within normal limits351-07MSMacroscopicexternal appearance- within normal limits351-08MSMacroscopicexternal appearance- within normal limits351-09MSMacroscopicexternal appearance- within normal limits351-10MSMacroscopicexternal appearance- within normal limits351-11MSMacroscopicexternal appearance- within normal limits 

# Page 304 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed351-12MSMacroscopicexternal appearance- within normal limits351-13FSMacroscopicexternal appearance- within normal limits351-14FSMacroscopicexternal appearance- within normal limits351-15FSMacroscopicexternal appearance- within normal limits351-16FSMacroscopicexternal appearance- within normal limits351-17FSMacroscopicexternal appearance- within normal limits353-01MSMacroscopicexternal appearance- within normal limits353-02MSMacroscopicexternal appearance- within normal limits353-03MSMacroscopicexternal appearance- within normal limits353-04MSMacroscopicexternal appearance- within normal limits 

# Page 305 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed353-05MSMacroscopicexternal appearance- within normal limits353-06MSMacroscopicexternal appearance- within normal limits353-07MSMacroscopicexternal appearance- within normal limits353-08MSMacroscopicexternal appearance- within normal limits353-12FSMacroscopicexternal appearance- within normal limits353-13FSMacroscopicexternal appearance- within normal limits353-14FSMacroscopicexternal appearance- within normal limits353-15FSMacroscopicexternal appearance- within normal limits353-16FSMacroscopicexternal appearance- within normal limits354-01MSMacroscopicexternal appearance- within normal limits 

# Page 306 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed354-02MSMacroscopicexternal appearance- within normal limits354-03MSMacroscopicexternal appearance- within normal limits354-04MSMacroscopicexternal appearance- within normal limits354-05MSMacroscopicexternal appearance- within normal limits354-06MSMacroscopicexternal appearance- within normal limits354-07MSMacroscopicexternal appearance- within normal limits354-09FSMacroscopicexternal appearance- within normal limits354-10FSMacroscopicexternal appearance- within normal limits354-11FSMacroscopicexternal appearance- within normal limits354-12FSMacroscopicexternal appearance- within normal limits 

# Page 307 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed354-13FSMacroscopicexternal appearance- within normal limits354-14FSMacroscopicexternal appearance- within normal limits354-15FSMacroscopicexternal appearance- within normal limits354-16FSMacroscopicexternal appearance- within normal limits355-01MSMacroscopicexternal appearance- within normal limits355-02MSMacroscopicexternal appearance- within normal limits355-03MSMacroscopicexternal appearance- within normal limits355-04MSMacroscopicexternal appearance- within normal limits355-05MSMacroscopicexternal appearance- within normal limits355-06MSMacroscopicexternal appearance- within normal limits 

# Page 308 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed355-07MSMacroscopicexternal appearance- within normal limits355-08MSMacroscopicexternal appearance- within normal limits355-09FSMacroscopicexternal appearance- within normal limits355-10FSMacroscopicexternal appearance- within normal limits355-11FSMacroscopicexternal appearance- within normal limits355-12FSMacroscopicexternal appearance- within normal limits355-13FSMacroscopicexternal appearance- within normal limits356-01MSMacroscopicexternal appearance- within normal limits356-02MSMacroscopicexternal appearance- within normal limits356-03MSMacroscopicexternal appearance- within normal limits 

# Page 309 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed356-04MSMacroscopicexternal appearance- within normal limits356-05MSMacroscopicexternal appearance- within normal limits356-06MSMacroscopicexternal appearance- within normal limits356-07MSMacroscopicexternal appearance- within normal limits356-08MSMacroscopicexternal appearance- within normal limits356-09MSMacroscopicexternal appearance- within normal limits356-10MSMacroscopicexternal appearance- within normal limits356-11MSMacroscopicexternal appearance- within normal limits356-12FSMacroscopicexternal appearance- within normal limits356-13FSMacroscopicexternal appearance- within normal limits 

# Page 310 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed356-14FSMacroscopicexternal appearance- within normal limits356-16FSMacroscopicexternal appearance- within normal limits356-17FSMacroscopicexternal appearance- within normal limits357-01MSMacroscopicexternal appearance- within normal limits357-02MSMacroscopicexternal appearance- within normal limits357-03MSMacroscopicexternal appearance- within normal limits357-04MSMacroscopicexternal appearance- within normal limits357-05FSMacroscopicexternal appearance- within normal limits357-06FSMacroscopicexternal appearance- within normal limits357-07FSMacroscopicexternal appearance- within normal limits 

# Page 311 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed357-08FSMacroscopicexternal appearance- within normal limits357-09FSMacroscopicexternal appearance- within normal limits357-10FSMacroscopicexternal appearance- within normal limits357-11FSMacroscopicexternal appearance- within normal limits357-12FSMacroscopicexternal appearance- within normal limits357-13FSMacroscopicexternal appearance- within normal limits358-01MSMacroscopicexternal appearance- within normal limits358-02MSMacroscopicexternal appearance- within normal limits358-03MSMacroscopicexternal appearance- within normal limits358-04MSMacroscopicexternal appearance- within normal limits 

# Page 312 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed358-05MSMacroscopicexternal appearance- within normal limits358-06MSMacroscopicexternal appearance- within normal limits358-07MSMacroscopicexternal appearance- within normal limits358-09FSMacroscopicexternal appearance- within normal limits358-10FSMacroscopicexternal appearance- within normal limits358-11FSMacroscopicexternal appearance- within normal limits358-12FSMacroscopicexternal appearance- within normal limits358-13FSMacroscopicexternal appearance- within normal limits359-01MSMacroscopicexternal appearance- within normal limits359-02MSMacroscopicexternal appearance- within normal limits 

# Page 313 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed359-03MSMacroscopicexternal appearance- within normal limits359-04MSMacroscopicexternal appearance- within normal limits359-05MSMacroscopicexternal appearance- within normal limits359-06FSMacroscopicexternal appearance- within normal limits359-07FSMacroscopicexternal appearance- within normal limits359-08FSMacroscopicexternal appearance- within normal limits359-09FSMacroscopicexternal appearance- within normal limits359-10FSMacroscopicexternal appearance- within normal limits359-11FSMacroscopicexternal appearance- within normal limits359-12FSMacroscopicexternal appearance- within normal limits 

# Page 314 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed360-01MSMacroscopicexternal appearance- within normal limits360-02MSMacroscopicexternal appearance- within normal limits360-03MSMacroscopicexternal appearance- within normal limits360-04MSMacroscopicexternal appearance- within normal limits360-05MSMacroscopicexternal appearance- within normal limits360-06MSMacroscopicexternal appearance- within normal limits360-07MSMacroscopicexternal appearance- within normal limits360-08MSMacroscopicexternal appearance- within normal limits360-09MSMacroscopicexternal appearance- within normal limits 

# Page 315 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations0 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed360-10MSMacroscopicexternal appearance- within normal limits360-11MSMacroscopicexternal appearance- within normal limits360-12MSMacroscopicexternal appearance- within normal limits360-13MSMacroscopicexternal appearance- within normal limits360-14MSMacroscopicexternal appearance- within normal limits360-15FSMacroscopicexternal appearance- within normal limits360-16FSMacroscopicexternal appearance- within normal limits 

# Page 316 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed362-01MSMacroscopicexternal appearance- within normal limits362-02MSMacroscopicexternal appearance- within normal limits362-03MSMacroscopicexternal appearance- within normal limits362-04FSMacroscopicexternal appearance- within normal limits362-05FSMacroscopicexternal appearance- within normal limits362-06FSMacroscopicexternal appearance- within normal limits362-07FSMacroscopicexternal appearance- within normal limits362-08FSMacroscopicexternal appearance- within normal limits362-09FSMacroscopicexternal appearance- within normal limits362-10FSMacroscopicexternal appearance- within normal limits 

# Page 317 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed363-01MSMacroscopicexternal appearance- within normal limits363-02MSMacroscopicexternal appearance- within normal limits363-03MSMacroscopicexternal appearance- within normal limits363-04MSMacroscopicexternal appearance- within normal limits363-05MSMacroscopicexternal appearance- within normal limits363-06FSMacroscopicexternal appearance- within normal limits363-07FSMacroscopicexternal appearance- within normal limits363-08FSMacroscopicexternal appearance- within normal limits363-09FSMacroscopicexternal appearance- within normal limits363-10FSMacroscopicexternal appearance- within normal limits 

# Page 318 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed363-11FSMacroscopicexternal appearance- within normal limits363-12FSMacroscopicexternal appearance- within normal limits364-01MSMacroscopicexternal appearance- within normal limits364-02MSMacroscopicexternal appearance- within normal limits364-03MSMacroscopicexternal appearance- within normal limits364-04MSMacroscopicexternal appearance- within normal limits364-05MSMacroscopicexternal appearance- within normal limits364-06MSMacroscopicexternal appearance- within normal limits364-07MSMacroscopicexternal appearance- within normal limits364-08MSMacroscopicexternal appearance- within normal limits 

# Page 319 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed364-09MSMacroscopicexternal appearance- within normal limits364-10MSMacroscopicexternal appearance- within normal limits364-11FSMacroscopicexternal appearance- within normal limits364-12FSMacroscopicexternal appearance- within normal limits364-13FSMacroscopicexternal appearance- within normal limits365-01MSMacroscopicexternal appearance- within normal limits365-02MSMacroscopicexternal appearance- within normal limits365-03MSMacroscopicexternal appearance- within normal limits365-04MSMacroscopicexternal appearance- within normal limits365-05FSMacroscopicexternal appearance- within normal limits 

# Page 320 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed365-06FSMacroscopicexternal appearance- within normal limits365-07FSMacroscopicexternal appearance- within normal limits365-08FSMacroscopicexternal appearance- within normal limits365-09FSMacroscopicexternal appearance- within normal limits365-10FSMacroscopicexternal appearance- within normal limits365-11FSMacroscopicexternal appearance- within normal limits365-12FSMacroscopicexternal appearance- within normal limits365-13FSMacroscopicexternal appearance- within normal limits365-14FSMacroscopicexternal appearance- within normal limits366-01MSMacroscopicexternal appearance- within normal limits 

# Page 321 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed366-02MSMacroscopicexternal appearance- within normal limits366-03MSMacroscopicexternal appearance- within normal limits366-04MSMacroscopicexternal appearance- within normal limits366-05MSMacroscopicexternal appearance- within normal limits366-06FSMacroscopicexternal appearance- within normal limits366-07FSMacroscopicexternal appearance- within normal limits366-08FSMacroscopicexternal appearance- within normal limits366-09FSMacroscopicexternal appearance- within normal limits366-10FSMacroscopicexternal appearance- within normal limits366-11FSMacroscopicexternal appearance- within normal limits 

# Page 322 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed366-12FSMacroscopicexternal appearance- within normal limits367-01MSMacroscopicexternal appearance- within normal limits367-02MSMacroscopicexternal appearance- within normal limits367-03MSMacroscopicexternal appearance- within normal limits367-04MSMacroscopicexternal appearance- within normal limits367-05MSMacroscopicexternal appearance- within normal limits367-06MSMacroscopicexternal appearance- within normal limits367-07MSMacroscopicexternal appearance- within normal limits367-08FSMacroscopicexternal appearance- within normal limits367-09FSMacroscopicexternal appearance- within normal limits 

# Page 323 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed367-10FSMacroscopicexternal appearance- within normal limits367-11FSMacroscopicexternal appearance- within normal limits367-12FSMacroscopicexternal appearance- within normal limits367-13FSMacroscopicexternal appearance- within normal limits367-14FSMacroscopicexternal appearance- within normal limits368-01MSMacroscopicexternal appearance- within normal limits368-02MSMacroscopicexternal appearance- within normal limits368-03MSMacroscopicexternal appearance- within normal limits368-04MSMacroscopicexternal appearance- within normal limits368-05MSMacroscopicexternal appearance- within normal limits 

# Page 324 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed368-06FSMacroscopicexternal appearance- within normal limits368-07FSMacroscopicexternal appearance- within normal limits368-09FSMacroscopicexternal appearance- within normal limits368-10FSMacroscopicexternal appearance- within normal limits369-01MSMacroscopicexternal appearance- within normal limits369-02MSMacroscopicexternal appearance- within normal limits369-03MSMacroscopicexternal appearance- within normal limits369-04MSMacroscopicexternal appearance- within normal limits369-05MSMacroscopicexternal appearance- within normal limits369-06MSMacroscopicexternal appearance- within normal limits 

# Page 325 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed369-07MSMacroscopicexternal appearance- within normal limits369-08FSMacroscopicexternal appearance- within normal limits369-09FSMacroscopicexternal appearance- within normal limits369-10FSMacroscopicexternal appearance- within normal limits369-11FSMacroscopicexternal appearance- within normal limits369-12FSMacroscopicexternal appearance- within normal limits369-13FSMacroscopicexternal appearance- within normal limits369-14FSMacroscopicexternal appearance- within normal limits369-15FSMacroscopicexternal appearance- within normal limits370-01MSMacroscopicexternal appearance- within normal limits 

# Page 326 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed370-02MSMacroscopicexternal appearance- within normal limits370-03MSMacroscopicexternal appearance- within normal limits370-04MSMacroscopicexternal appearance- within normal limits370-05MSMacroscopicexternal appearance- within normal limits370-06FSMacroscopicexternal appearance- within normal limits370-07FSMacroscopicexternal appearance- within normal limits370-08FSMacroscopicexternal appearance- within normal limits370-09FSMacroscopicexternal appearance- within normal limits370-10FSMacroscopicexternal appearance- within normal limits370-11FSMacroscopicexternal appearance- within normal limits 

# Page 327 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations150 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed370-12FSMacroscopicexternal appearance- within normal limits370-13FSMacroscopicexternal appearance- within normal limits 

# Page 328 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations600 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed383-01MSMacroscopicexternal appearance- within normal limits383-02MSMacroscopicexternal appearance- within normal limits383-05FSMacroscopicexternal appearance- within normal limits383-06FSMacroscopicexternal appearance- within normal limits383-07FSMacroscopicexternal appearance- within normal limits383-09FSMacroscopicexternal appearance- within normal limits383-10FSMacroscopicexternal appearance- within normal limits383-11FSMacroscopicexternal appearance- within normal limits383-12FSMacroscopicexternal appearance- within normal limits384-12FSMacroscopicexternal appearance- within normal limits 

# Page 329 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed389-01MSMacroscopicexternal appearance- within normal limits389-02MSMacroscopicexternal appearance- within normal limits389-03MSMacroscopicexternal appearance- within normal limits389-04FSMacroscopicexternal appearance- within normal limits389-05FSMacroscopicexternal appearance- within normal limits389-06FSMacroscopicexternal appearance- within normal limits389-07FSMacroscopicexternal appearance- within normal limits389-08FSMacroscopicexternal appearance- within normal limits389-09FSMacroscopicexternal appearance- within normal limits389-10FSMacroscopicexternal appearance- within normal limits 

# Page 330 of 358MPI Research Study Number 1271-001An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CAS # 84-65-1) in RatsIndividual F1Pup Animal Data Record: PathologyDay 4 PupsGroup,Animal NumberSexFate TissueObservations2400 mg/kg/dayS - Scheduled Necropsy E - Euthanizedin extremisD - Died on Study U – Unable to Determine Sex NT – Not Tattooed389-11FSMacroscopicexternal appearance- within normal limits389-12FSMacroscopicexternal appearance- within normal limits 

# Page 331 of 358

# 1271-001 Appendix R

Protocol and Amendments Page 332 of 358

# AN ORAL REPRODUCTION/DEVELOPMENTAL TOXICITY

SCREENING STUDY OF ANTHRAQUINO NE (CAS # 84-65-1) IN RATS

TESTING FACILITY

MPI Research, Inc.

54943 North Main Street

Mattawan, MI 49071-9399 U.S.A.



# STUDY NUMBER

1271-001



# STUDY DIRECTOR

Cheryl K. Lanphear, B.S.



# SPONSOR

Chemical Products Corporation

102 Old Mill Road, SE

P.O. Box 2470

Cartersville, GA 30120-1692

June 5, 2006 Page 333 of 358

# MPI Research Study Number: 1271-001

2 Table of Contents

1. INTRODUCTION ..................................................................................................... 4

1.1. Study Number ..................................................................................................... 4

1.2. Study Title........................................................................................................... 4

1.3. Sponsor................................................................................................................ 4

1.4. Sponsor Representative....................................................................................... 4

1.5. Objective ............................................................................................................. 4

1.6. Regulatory Guideline .......................................................................................... 4

1.7. Good Laboratory Practice ................................................................................... 4

1.8. Testing Facility.................................................................................................... 4

1.9. Computer Systems............................................................................................... 5

1.10. Personnel ............................................................................................................. 5

1.11. Proposed Study Schedule.................................................................................... 6

1.12. Quality Assurance ............................................................................................... 6

1.13. Alteration of Design............................................................................................ 6

1.14. Declaration of Intent............................................................................................ 6

2. TEST AND CONTROL ARTICLES ........................................................................ 6

2.1. Description of Test Article.................................................................................. 6

2.2. Test Article Preparation ...................................................................................... 6

2.3. Test Article Analysis........................................................................................... 7

2.4. Reserve Sample................................................................................................... 7

2.5. Test Article Disposition ...................................................................................... 7

2.6. Description of Vehicle/Control ........................................................................... 8

3. TEST SYSTEM ......................................................................................................... 8

3.1. Species................................................................................................................. 8

3.2. Strain ................................................................................................................... 8

3.3. Source.................................................................................................................. 8

3.4. Justification of Test System ................................................................................ 8

3.5. Expected Age ...................................................................................................... 8

3.6. Expected Body Weight........................................................................................ 9

3.7. Number of Animals............................................................................................. 9

3.8. Husbandry ........................................................................................................... 9

4. STUDY DESIGN..................................................................................................... 11

5. TEST AND CONTROL ARTICLE ADMINISTRATION..................................... 11

5.1. Route of Administration.................................................................................... 11

5.2. Justification for Route of Administration.......................................................... 11

5.3. Frequency of Administration ............................................................................ 11

5.4. Duration of Administration ............................................................................... 11

5.5. Dose Levels....................................................................................................... 11

5.6. Dose Volume..................................................................................................... 11

5.7. Justification of Dose Levels .............................................................................. 11

5.8. Test and Control Article Administration........................................................... 11

6. ANTEMORTEM STUDY EVALUATIONS.......................................................... 12

6.1. Cageside Observations ...................................................................................... 12 Page 334 of 358

# MPI Research Study Number: 1271-001

3 6.2. Detailed Clinical Observations.......................................................................... 12

6.3. Body Weights.................................................................................................... 12

6.4. Food Consumption ............................................................................................ 12

6.5. Estrous Cyclicity ............................................................................................... 12

6.6. P Breeding Procedures ...................................................................................... 12

6.7. P Parturition and F 1 

# Litter Observations ........................................................... 12

7. EUTHANASIA........................................................................................................ 13

7.1. Moribundity....................................................................................................... 13

7.2. Method of Euthanasia........................................................................................ 13

7.3. Final Disposition ............................................................................................... 13

8. POSTMORTEM EVALUATIONS......................................................................... 13

8.1. Anatomic Pathology.......................................................................................... 13

9. STATISTICS ........................................................................................................... 15

9.1. Group Pair-Wise Comparisons (L evene’s/ANOVA-Dunnett’s/Welch’s) ........ 16

9.2. Arcsin-Square-Root Transforma tion Followed by Group-Pairwise ................. 16

9.3. Fisher’s Exact Test............................................................................................ 17

9.4. Covariate Analysis ............................................................................................ 17

10. STUDY REPORTS.................................................................................................. 17

10.1. Progress/Status Reports..................................................................................... 17

10.2. Final Report....................................................................................................... 17

11. DATA AND SPECIMEN RETENTION................................................................. 18

12. ANIMAL WELFARE.............................................................................................. 18

13. SIGNATURES......................................................................................................... 19

13.1. Sponsor Approval.............................................................................................. 19

13.2. Study Director Approval/Study Initiation......................................................... 19 Page 335 of 358

# MPI Research Study Number: 1271-001

4 1. INTRODUCTION

1.1. Study Number

1271-001

1.2. Study Title

An Oral Reproduction/Developmental Toxicity Screeni ng Study of Anthraquinone

(CAS # 84-65-1) in Rats

1.3. Sponsor

Chemical Products Corporation

102 Old Mill Road, SE

P.O. Box 2470

Cartersville, GA 30120-1692

1.4. Sponsor Representative

Jerry A. Cook, Technical Director

Telephone: 770-382-2144

Telefax: 770-386-6053

E-mail: jcook@cpc-us.com

1.5. Objective

The objective of this study is to evaluate the effects of the te st article on male and female

reproductive performance, including gonadal function, estrous cycle, mating behavior,

conception, development of the concep tus, and parturition in rats.

1.6. Regulatory Guideline

This protocol meets or exceeds Guideline 421, Reproduction/Developmental Toxicity Screening

Test, the Organization of Economic Cooperatio n and Development (OECD) Guideline for

Testing of Chemicals, revised July 1995.

1.7. Good Laboratory Practice

This nonclinical laboratory study will be conduc ted in accordance with the United States

Environmental Protection Agency (EPA), T oxic Substance Control Act (TSCA) Good

Laboratory Practice (GLP) Standards, 40 CF R Part 792, effective September 18, 1989, and

OECD Principles of GLP ENV/MC/CHEM(98)17.

1.8. Testing Facility

MPI Research, Inc.

54943 North Main Street

Mattawan, MI 49071-9399 U.S.A.

MPI Research is fully accredited by the Asso ciation for Assessment and Accreditation of

Laboratory Animal Care Internat ional (AAALAC International). Page 336 of 358

# MPI Research Study Number: 1271-001

5 1.9. Computer Systems

The following are the proposed computer systems to be used during the conduct of this study.

The actual systems used will be documented in the final report.

MPI Research Computer Systems

Provantis TM 

# : Client-server, Oracle-based system primarily used for

toxicology studies.

Niagara Framework ® 

# Software System: Environmental monitoring, alarming, and reporting

application.

Dispense: Automates the test article control processes.

Millennium 32 

# : Millennium 32 

# HPLC System (instrument) and

Millennium 32 

# Chromatography Manager Version

(software) used to quantitatively determine the amount

of test article in prepar ed test article batches.

MPI Archiving System (MArcS): In-house deve loped application for automated storage

and retrieval information for archiveable materials

(e.g. lab books, study data, wet tissues, slides, etc.).

MPI Reporting System (MPIRS): In-house developed reporting system running SAS ®



# programs primarily reporting on Provantis TM 

# data.

Master Schedule: Maintains the ma ster schedule for the company.

SAS ® 

# : The SAS ® 

# System is an integrated system of software

products that enables a user to perform data entry,

retrieval, data management, reporting, graphics,

statistical analysis, and a pplications development.

Microsoft ® 

# Office 2003 Professional: Bundle of integrated produc tivity tools including word

and data processing and co mmunications software.

Contains the utilities Microsoft ® 

# Access, Excel,

InfoPath, Outlook, PowerPoint, Publisher, and Word.

1.10. Personnel

1.10.1. Study Director

Cheryl K. Lanphear, B.S.

Telephone: 269-668-3336 ext. 1450

Telefax: 269-668-4151

E-mail: cheryl.lanphear@mpiresearch.com

1.10.2. Alternate Contact

Raymond E. Schroeder, M.S., D.A.B.T. ®



# Telephone: 269-668-3336 ext. 1215

Telefax: 269-668-4151

E-mail: ray.schroeder@mpiresearch.com Page 337 of 358

# MPI Research Study Number: 1271-001

6 1.11. Proposed Study Schedule

Study Initiation Date: Date St udy Director signs Study ApprovalInitiation Line in this protocol

Experimental Starting Date (OECD): To be added by amendment

Experimental Start Date: To be added by amendment

Experimental Termination Date: To be added by amendment

Experimental Completion Date (OECD): To be added by amendment.

Draft Report Mail Date: To be added by amendment

1.12. Quality Assurance

This study will be subjected to periodic inspect ions and the draft and final reports will be

reviewed by the Quality Assurance Departme nt of MPI Research in accordance with MPI

Research’s Standard Operating Procedures. St udy quality assurance inspection records will be

made available to the Sponsor Represen tatives during visits to MPI Research.

1.13. Alteration of Design

Alterations of this protocol may be made as th e study progresses. No ch anges in the protocol

will be made without the specific wr itten request or consent of the Sponsor. In the event that the

Sponsor authorizes a protocol change verbally, MPI Research will honor such change. However,

written authorization will be obtained thereafter. All protocol modifications and the reasons will

be documented, signed, and dated by the Study Di rector and Sponsor. The protocol and all

amendments will be issued to the Sponsor as well as at MPI Research.

1.14. Declaration of Intent

This study may be submitted to the United States Environmental Protection Agency (EPA) and

an Organization for Economic Cooperation a nd Development (OECD) member country.

2. TEST AND CONTROL ARTICLES

2.1. Description of Test Article

2.1.1. Identity

Anthraquinone (CAS # 84-65-1)

A description, lot number, storag e conditions, expiration date, safe handling procedures, physical

properties, as well as other relevant info rmation will be documented in the study data.

2.1.2. Test Article Properties

The Sponsor will provide documentation on the strength, purity, composition, stability, and other

pertinent information on each batch of test artic le, unless otherwise noted. If the Sponsor does

not supply the above information (e.g. certificate of analysis), this will be listed as a GLP

deviation in the final report.

2.2. Test Article Preparation

The test article will be used as received from the Sponsor and no adjustment will be made for

purity. The test article will be d iluted with an appropriate vehicle in order to achieve the desired Page 338 of 358

# MPI Research Study Number: 1271-001

7 dose volumes. If the material is a solid/powder, appropriate amounts of th e test article will be

mixed with an appropriate vehi cle to achieve the desired conc entrations. The vehicle and

method of preparation will be determined based u pon physical characteristic s of the test article

and size of batches required. Fresh suspensions will be prepared for each concentration weekly,

unless the stability analysis results indicate otherwis e. Preparations will be stored refrigerated

until used. Each daily aliquot will be removed from the refrigerator and stirred continuously

until completion of dosing.

2.3. Test Article Analysis

2.3.1. Homogeneity

To comply with the regulatory requirement to document test article homogeneity in all

formulations, prior to initiation of test article administration, te st batches of the test article

formulation at both the low and high concentrati ons will be prepared and analyzed. The test

batches will be prepared employing the same met hod and batch sizes to be used during the study.

Following mixing, duplicate 2 mL samples (top, middle, and bottom) will be collected and stored

refrigerated until analyzed. Remixing may be necessary to obtain satisfactory homogeneity

results. Following completion of the study any remaining samples will be properly disposed.

2.3.2. Stability Analysis

Appropriately sized portions of the prepared su spensions mixed for the homogeneity analysis

will be stored under refrigerated conditions for up to 14 days. Samples (2 mL) of these stored

preparations will be collected after 7 and 14 days at refrigerated temperature and analyzed to

determine the stability of the test article in the vehicle. If the result s of this analysis are

unsatisfactory, further testing may be necessary to obtain satisfactory stabil ity results. Following

completion of the study any remaining samples will be properly disposed.

2.3.3. Concentration Analysis

Samples (2 mL) of the dosing formulations at each concentration will be collected from the first,

third, and last preparation for an alysis of test article concentr ation. Samples will be stored

refrigerated until analyzed. Following completi on of the study any remaining samples will be

properly disposed.

2.3.4. Analyses

All analytical work will be conducted by MPI Research, Inc. using an analytical method

developed and validated under MP I Research Study Number 1271-002.

2.4. Reserve Sample

A reserve sample from each batch of a test article used in this study will be taken and archived at

MPI Research. If multipl e studies are conducted with the same test article, a common reserve

sample may be taken an d labeled appropriately.

2.5. Test Article Disposition

Any remaining test article will be shipped to a Sponsor-designated location after completion of

the study. The Sponsor will be notified prior to such shipment. Alternatively, MPI Research

will dispose of the test ar ticle after completion of the study (additional cost). Page 339 of 358

# MPI Research Study Number: 1271-001

8 2.6. Description of Vehicle/Control

2.6.1. Identity

1% Guar Gum in distilled water (distilled from deionized tap water at the Testing Facility).

A description, lot number, storag e conditions, expiration date, safe handling procedures, physical

properties, as well as other relevant info rmation will be documented in the study data.

2.6.2. Vehicle/Control Properties

Documentation of the strength, purity, composition, stability, and other pertinent information on

each batch of vehicle, will be limited to that in formation listed on the label of this commercially

available vehicle, unless otherwise noted.

3. TEST SYSTEM

3.1. Species

Rat

3.2. Strain

CD ® 

# [Crl:CD ® 

# (SD)]

3.3. Source

Charles River Laboratories

3.4. Justification of Test System

The current state of scientific knowledge and the a pplicable guidelines cited previously in this

protocol does not provide acceptable alternatives, in vitro or otherwise, to the use of live animals

to accomplish the purpose of this study. “The development of knowledge necessary for the

improvement of the health and well-being of humans as well as ot her animals requires in vivo

experimentation with a wide variety of animal species". 1 

# "Whole animals are essential in

research and testing because they best reflect the dynamic interactions be tween the various cells,

tissues, and organs co mprising the human body". 2



# The rat is the usual rodent model 3 

# used for evaluating the toxicity of various classes of chemicals

and for which there is a la rge historical database.

3.5. Expected Age

The test animals will be approximately 6 weeks of age at arrival.

1 

# “Principles for the Utilization and Care of Vertebrate An imals Used in Testing, Research, and Training”, Federal

Register, Vol. 50, No. 97, May 20, 1985

2 

# “Position Statement on the Use of Animals in Research”, NIH Guide 22: No. 8, Feb.26, 1993

3 

# Guidance for Industry, Investigators, and Reviewers: Exploratory IND Studies, U.S. F.D.A. Center for Drug

Evaluation and Research (CDER), January 2006. 

# Page 340 of 358

# MPI Research Study Number: 1271-001

9 3.6. Expected Body Weight

The males will weigh approximately 130 to 210 g and females will weigh approximately 100 to

170 g at arrival, as measured within 3 days of arrival. The actual range may vary but will be

documented in the data.

3.7. Number of Animals

3.7.1. Number Ordered

Males: 58

Females: 58

3.7.2. Number on Study

Males: 48

Females: 48

Females will be nulliparous and nonpregnant

F 1 

# pups (based on approximately 14 pups per litte r): approximately 672 pups (actual number

will be documented in the study data).

3.7.3. Justification for Number on Study

This study was designed to use th e fewest number of animals po ssible, consistent with the

objective of the study, the sc ientific needs of the Sponsor, contemporary scie ntific standards, and

in consideration of applicable regulatory requirements cited previously in this protocol.

3.7.4. Selection for Study

All animals placed on study will have body weights that fall within ± 20% of the mean body

weight for each sex. If not enough animals fall with in this weight range to satisfy the number of

animals required to go on study, the Study Director will be notified to as certain the appropriate

action to be taken.

Animals considered suitable for study will be wei ghed prior to treatment. After the appropriate

number of animals with the highest and lowest body weights has been excluded, the remaining

required number of animals on study will be rand omized, by sex, into treatment groups using a

standard, by weight, bloc k randomization procedure.

3.7.5. Method of Identification

Each animal will be assigned an animal number to be used in Provantis TM 

# and will be implanted

with a microchip bearing a unique identificati on number. Pups will be identified by tattoo on

approximately Lactation Day (LD) 0. The indi vidual animal number, implant number, and the

MPI Research study number will comprise a unique identification for each animal. The animal

cage will be identified by the study numb er, animal number, group number, and sex.

3.8. Husbandry

3.8.1. Acclimation

All animals will be permitted an acclimation period of at least 2 weeks. During this acclimation

period, all animals will be observed daily for any clin ical signs of disease, and all animals will be Page 341 of 358

# MPI Research Study Number: 1271-001

10 given a detailed clinical examination prior to se lection for study. All animals with any evidence

of disease or physical abnormalities will not be se lected for study. Animals considered suitable

for study will be randomly selected and divided into groups as described above.

3.8.2. Housing

The animals will be individually housed in suspe nded, stainless steel, wire-mesh type cages,

except during pairing, near parturition, and durin g lactation. During pairing, animals will be

housed 2/cage. On approximately Gestation Da y (GD) 20, females will be individually housed

in plastic cages containing wood ch ip bedding until euthanasia on LD 4.

3.8.3. Environmental Conditions

Fluorescent lighting will be provide d via an automatic timer for approximately 12 hours per day.

On occasion, the dark cycle may be interrupted intermittently due to study-related activities.

Temperature and humidity will be monitored and r ecorded daily and maintained to the maximum

extent possible between 64 to 79° F and 30 to 70%, respectively.

3.8.4. Diet and Drinking Water

3.8.4.1. Basal Diet

The basal diet will be meal Lab Diet ® 

# Certified Rodent Diet #5002, PM I Nutrition International,

Inc. This diet will be available ad libitum unless designated otherwise. Each lot number used

will be identified in the study records.

3.8.4.2. Basal Diet Contaminants

The Study Director is not aware of any potential contaminants likel y to be present in the certified

diet that would interfere with th e results of this study. Therefor e, no analyses other than those

routinely performed by the f eed supplier will be conducted.

3.8.4.3. Water

Tap water will be supplied ad libitum to all animals via an automatic water system unless

otherwise indicated.

3.8.4.4. Water Contaminants

The drinking water used will be monitored for specified contaminants at periodic intervals

according to MPI Research Standard Operating Pr ocedures. The Study Director is not aware of

any potential contaminants likely to be present in the water that would interfere with the results

of this study. Therefore, no analyses other th an those mentioned in this protocol will be

conducted. Page 342 of 358

# MPI Research Study Number: 1271-001

11 4. STUDY DESIGN

NUMBER OF ANIMALS

(P GENERATION)

DOSE

LEVEL INITIAL NECROPSY

GROUP mg/kg/day M F M F

1 0 12 12 12 12

2 150 12 12 12 12

3 600 12 12 12 12

4 2400 12 12 12 12



# 5. TEST AND CONTROL ARTICLE ADMINISTRATION

5.1. Route of Administration

The test and control articles wi ll be administ ered by gavage.

5.2. Justification for Route of Administration

The oral route is one of the potential routes of human e xposure to this test article.

5.3. Frequency of Administration

The test and control articles will be administered once per day.

5.4. Duration of Administration

Dosing of the males will begin 14 days prior to ma ting and continue to terminal euthanasia (14

day mating period and 14 days post-mating). Dosi ng of the females will begin 14 days prior to

mating and continue through LD 4. Unmated fe males will continue to be dosed following

completion of the mating period until determined not to be pregnant. Dosing will be performed

at approximately the same time each day (± 2 hours from Day 1).

5.5. Dose Levels

0, 150, 600, and 2400 mg per kg per day.

5.6. Dose Volume

20 mL per kg per dose

5.7. Justification of Dose Levels

The dose levels were selected by the Sponsor, or in consultation with the Sponsor, on the basis of

available data from previous studies.

5.8. Test and Control Article Administration

The test and control articles will be administer ed via gavage. The te st and control article

preparations will be stirred continuously through out dosing. The control an imals will receive the

control article at the same volume as the test articl e. Individual doses will be based on the most

recent body weights. Page 343 of 358

# MPI Research Study Number: 1271-001

12 6. ANTEMORTEM STUDY EVALUATIONS

6.1. Cageside Observations

All rats will be observed at le ast twice a day, 7 days a week, fo r morbidity, mortality, and signs

of injury. Any findings will be recorded. S hould mortality or other signs of toxicity be

observed, these findings will be recorded on the day they are observed.

6.2. Detailed Clinical Observations

A detailed clinical observation of each animal will be performed daily during treatment (30-90

minutes postdose). The examination will include, but will not be limited to, evaluation of the

skin, fur, eyes, ears, nose, oral cavity, thorax, ab domen, external genitalia, limbs and feet, as well

as evaluation of respiration.

6.3. Body Weights

Individual body weights will be measured and recorded weekly during the study. All females

will also be weighed on GD 0, 7, 14, and 20 and on LD 0 and 4.

6.4. Food Consumption

Individual food consumption will be measured and recorded weekly during the study, except

during the mating period. Duri ng the mating period, food consumption will not be measured

because the animals will be coha bitating at this time. Food cons umption will also be measured

on GD 0, 7, 14, and 20 and on LD 0 and 4.

6.5. Estrous Cyclicity

At initiation of test article ad ministration and until evidence of copulation is observed or the

cohabitation period has ended, the females will be examined daily to establish estrous cycles.

Vaginal lavage will be completed daily by 10:00am.

6.6. P Breeding Procedures

The P animals will be individually housed until mating. After 2 weeks of treatment, the males

will be randomly cohabited, 1 male to 1 female , with the corresponding females in the treatment

and control groups. Mating will take place in the cage of the male.

Vaginal smears will be performed daily on female s during the mating period and the presence or

absence of sperm or vaginal plug will be reco rded. The day on which evidence of mating is

observed will be designated as GD 0. The rats will be paired a maximum of 14 days. If

evidence of mating is noted, the female will be removed from the cage of the male and

individually housed. After the mating period, an y female with no positive evidence of mating

will be individually housed in a plastic cage w ith wood chip bedding in anticipation of an

undetermined pregnancy.

After the mating period, the males will be individually housed in suspended, stainless-steel, wire

mesh type caging and continued on treatment for approximately 2 weeks. At the end of this

period, the males will be euthanized and necropsied according to the Anatomic Pathology

section.

6.7. P Parturition and F 1 

# Litter Observations

Toward the end of the gestation period, female s will be examined twice daily for signs of

parturition. The mated females w ill be allowed to give birth (F 1 

# ). The duration of gestation will Page 344 of 358

# MPI Research Study Number: 1271-001

13 be calculated, and any difficulties occurring at partur ition will be recorded. The day on which all

pups have been delivered will be designated as LD 0. The litters will be examined as soon as

possible after delivery, and the follo wing parameters will be recorded:

• Litter size

• Number of males and females

• Number of stillborn pups

• Number of live born pups

• Gross abnormalities of the pups

Litters will be weighed on LD 0 and 4, and any abnormal behavior observed in the pups will be

recorded daily. On LD 4, the pups will be eu thanized and evaluated as described in the

Anatomic Pathology section.

7. EUTHANASIA

7.1. Moribundity

Any moribund animals, as defined by the Testing F acility Standard Operating Procedure, will be

euthanized for humane reasons and to prevent the loss of tissues through autolysis. All animals

euthanized in extremis or found dead will be subjected to a routine necropsy. Where practical, a

full set of tissues as listed in the Postmortem Study Evaluations portion of this protocol will be

collected and preserved in the appropriate fixative.

7.2. Method of Euthanasia

Euthanasia will be by carbon dioxide inhalati on followed by an MPI Research SOP approved

method to ensure death, e.g. exsanguination. Euthan asia of fetuses/pups (LD 0-4) will be by an

intraperitoneal inj ection of sodium pe ntobarbital solution.

7.3. Final Disposition

All surviving animals placed on study will be euthanized at their scheduled necropsy or, if

necessary, euthanized in extremis . Extra animals obtained for this study, but not placed on study,

will be transferred to either an MPI Research stock or training colony, or euthanized and

discarded. The final disposition of each anim al will be documented in the study records.

8. POSTMORTEM EVALUATIONS

8.1. Anatomic Pathology

Complete necropsy examinations will be perfor med under procedures approved by a veterinary

pathologist on all adult animals dying spontaneously, euthanized in extremis , or terminated at

scheduled intervals. Adult animals that are found d ead will be necropsied as soon as possible. If

an animal cannot be necropsied immediately af ter being found (within 2 hours), the carcass will

be refrigerated until th e necropsy is performed.

At the termination of the study, all surviving adul t animals will be euthanized and examined. The

animals will be examined caref ully for external abnormalities including palpable masses. The

skin will be reflected from a ventral midline incision and any subcutaneous masses will be

identified and correlated with antemortem fi ndings. The abdominal, thoracic, and cranial

cavities will be examined for abnormalities. Sp ecial emphasis will be placed on organs of the Page 345 of 358

# MPI Research Study Number: 1271-001

14 reproductive system. Implantation sites will be c ounted and recorded in females. The organs

will be removed, examined and, where required by protocol, placed in neutral buffered formalin,

except the testes and epididymides which will be placed in Bouin’s fixative.

The body weight and testes, epididymides, seminal vesicles, and prostate weights of parental

males will be recorded. The left and right tes tis will be weighed separately, and a combined

weight will be presented in th e report. For processing of the Bouins-fixed testes, hematoxylin

and periodic acid Schiff (PAS) staining and pa raffin embedding will be performed. Plastic

(methacrylate) embedding of the testes and epididymides and staging of testes for

spermatogenesis can be conducted at th e Sponsor’s request (extra cost).

Body weight and the organ weight s identified in the following ta ble will be recorded for all

animals at scheduled necropsies and a ppropriate organ weight ratios will be calculated (relative

to body weights). Organs will not be weighed for animals dying spontaneously or euthanized in

extremis . The following tissues/organs from all animals will be removed, examined and placed

in neutral buffered formalin, except where spec ified otherwise. Micr oscopic examination of

formalin-fixed hematoxylin and eo sin-stained paraffin sections of the tissues indicated in the

table below will be examined.

Tissues to be Weighed, Preserved and Microscopically Examined

Organ

Weight Collected

and Microscopic

Examination

(Groups)

No. a 

# Tissue Taken Preserved 1, 4 2, 3

2 epididymis X X X

2 ovary X X X

1 prostate X X X

2 seminal vesicle X X X

2 testes X X X

1 uterus X X X

tissue masses with regional lymph

nodes X X X c



# gross lesions X X

identified target organs b 

# X X X c

a 

# Number of organs/sections preserved/examined.

b 

# Target organs (and target organ gross lesions) will be designated by the Study Director, Pathologist

and/or Sponsor based on experimental findings.

c 

# Any target organs examined in lower dose groups will incur additional cost.



# External examination of intact or aborted fetuse s (if present) from the animals listed above will

be conducted by Developmental Toxicology pers onnel under the supervisi on of a veterinary

pathologist, and the intact fetuses will be sa ved in 10% neutral buffere d formalin for possible

future examination.

F 1 

# pups from dams dying during la ctation will be externally examined, euthanized, and any

abnormalities will be recorded. The carcasses will then be discarded. Page 346 of 358

# MPI Research Study Number: 1271-001

15 On LD 4, offspring will be euthanized and ex ternally examined, and any abnormalities will be

recorded. The carcasses will then be discarde d. Any offspring found dead or euthanized in

extremis will be externally examined and discarded.

9. STATISTICS

The following is the proposed analysis plan to be used when data assumptions are met. If there

are deviations to this plan due to violations of assumptions or if any ot her techniques are used

(Sponsor consulted), they will be documented in the final report.

Table of Statistical Comparisons

Control Group Comparison Groups

1 2, 3, 4

The above table defines the set(s) of comparisons to be used in the statis tical analyses described

below. If more than one set of comparisons is required, all analyses wi ll be conducted separately

on each set unless stated otherwise. Data for each sex within a se t will be analyzed separately.

Endpoint Analysis

Parental In-life Data

Premating Body Weights Group Pair-wise Comparisons

Premating Food Consumption Group Pair-wise Comparisons

Gestation Body Weights Group Pair-wise Comparisons

Gestation Body Weight Cha nges Group Pair-wise Comparisons

Gestation Food Consumption Group Pair-wise Comparisons

Lactation Body Weights Group Pair-wise Comparisons

Lactation Body Weight Change s Group Pair-wise Comparisons

Lactation Food Consumption Group Pair-wise Comparisons

Postmating Body Weights-Male s Group Pair-wise Comparisons

Postmating Food Consumption-Ma les Group Pair-wise Comparisons

Fertility Indices

Estrous Cycle (mean cycle time) Group Pair-wise Comparisons

Estrous Cycle (# cycles/per iod) Group Pair-wise Comparisons

Gestation Length Gr oup Pair-wise Comparisons

Copulatory Interval Group Pair-wise Comparisons

Male Fertility Index Fisher's Exact Test

Female Fertility Index Fisher's Exact Test

Female Mating Index Fisher's Exact Test

Male Mating Index Fisher's Exact Test

Female Fecundity Index Fisher's Exact Test

Male Fecundity Index Fisher's Exact Test

Gestation Index Fisher's Exact Test Page 347 of 358

# MPI Research Study Number: 1271-001

16 Endpoint Analysis

Pathology

Male Reproduction Organ Wei ghts Group Pair-wise Comparisons

Female Reproduction Organ Wei ghts Group Pair-wise Comparisons

Litter

Litter Size Group Pair-wise Comparisons

Viable Pups Group Pair-wise Comparisons

Pup Sex Ratio (% viable males/litt er) Arcsin-Square-Root Transformation

Stillborn Pups Group Pair-wise Comparisons

Stillborn Index Arcsin -Square-Root Transformation

Pup Weights Covariate Analysis

Pup Survival (Days 0-4) Arcsin-Square-Root Transformation

9.1. Group Pair-Wise Comparisons (Levene’s/ANOVA-Dunnett’s/Welch’s)

If sample sizes for all groups are 3 or greater, Levene’s test 4 

# will be used to assess homogeneity

of group variances for each specified endpoint (see table above) and for all collection intervals. If

Levene’s test is not significant (p> 0.01), a pooled estimate of the variance (Mean Square Error or

MSE) will be computed from a one-way analys is of variance (ANOVA) and utilized by a

Dunnett’s 5 

# comparison of each treatment group with the control group. If Levene’s test is

significant (p<0.01), comparisons with the cont rol group will be made using Welch’s t-test 6 

# with

a Bonferroni correction.

In the case that sample size is less than 3 for at least one treatment group, Levene’s method

cannot be implemented. Groups with sample sizes less than 3 will be excluded from the analysis

and control-treatment pairwise comparisons that satisfy the sample size assumption (n> 3) will be

conducted using Welch’s t-test with a Bonferroni correction.

If there are only 2 groups involved, the above methodology applies and the Dunnett’s test

reduces to a Student’s t-test 7



# .

Results of all pair-wise compar isons will be reported at the 0. 05 and 0.01 significance levels. All

endpoints will be analyzed using two-taile d tests unless indicated otherwise.

9.2. Arcsin-Square-Root Transformation Followed by Group-Pairwise Comparisons

Data comprised of percent values will be tr ansformed using the arcsin of the square root 7 

# . The

analysis described in the Group Pair-Wise Comparison paragraph wi ll be used to analyze the

transformed percentage values.

4 

# Milliken GA, Johnson DE. Analysis of messy data. London: Chapman and Hall: 1992.

5 

# Dunnett CW. A multiple comparison procedure for comparin g several treatments with a control. J Am Stat Assoc

1955;50:1096-1121.

6 

# Welch BL. The significance of di fference between two means when the population variances are unequal.

Biometrika 1937;29:350-362.

7 

# Steel RGD, Torrie JH. Principles an d Procedures of Statistics. A biom etrical approach. New York: McGraw-Hill;

1980. 

# Page 348 of 358

# MPI Research Study Number: 1271-001

17 9.3. Fisher’s Exact Test

An overall test for association between response and treatment will be done using a Fisher’s

exact 8 

# test. If this overall test is significant (p < 0.05) and there are more than two groups, then a

follow up analysis will be done where each trea tment group will be compared to the control

group using a Fisher’s exact test. Results wi ll be reported at th e 0.05 and 0.01 significance

levels. All endpoints will be analyzed using two-tailed tests unless indicated otherwise.

Overall testing and follow up pair-w ise testing will be conducted fo r the control and all treatment

groups that have sufficient sample size (n ≥ 3). If all groups have an insufficient sample size

(n<3), or if just the control group has a sample size of less than 3, no testing will be performed.

9.4. Covariate Analysis

For groups with sample sizes greater than or equal to 3, an analysis of covariance 9 

# will be

conducted to compare treatment groups for each endpoint (mean of pup body weights per dam)

and time period. The covariate will be litter size. If the control group does not satisfy the sample

size assumption (n<3), no further analysis wi ll be performed. Each treatment satisfying the

sample size assumption will be compared to cont rol using Dunnett’s testunder the analysis of

covariance model.

The LSMEANS (Least Square Means), which are the means adjusted for values of the covariate,

will be displayed on the summary table.

Results of pairwise comparisons will be repo rted at the 0.05 and 0.01 si gnificance levels.

Endpoints will be analyzed using two-ta iled tests unless indicated otherwise.

10. STUDY REPORTS

10.1. Progress/Status Reports

Regular progress reports will be submitted to the Sponsor throughout the study. The frequency

of these reports will be determined after consultation with the Sponsor.

10.2. Final Report

After completion of the study, a comprehensive draf t report containing the results of all tests,

analyses, observations and measurements requi red by this protocol, and an interpretative

summary of the study results will be submitted to the Sponsor. The repor t will include all items

required by the applicable regulatory agency. Af ter receipt of any Sponsor comments, 2 copies

(1 bound and 1 unbound) of the final report will be issued.

Six months after issuance of the draft report, if no requested revi sions or instructions to finalize

have been communicated by the Sponsor, the draft re port will be issued as a final report, signed

by the Study Director, and submitted to the Sponsor . Any modifications or changes to the draft

report requested 6 months after issuance of the dr aft will be performed at additional cost to the

Sponsor.

8 

# Zar JH. Biostatistical Analysis. 4 th 

# ed. New Jersey: Prentice Hall; 1999.

9 

# Snedecor, GW, Cochran, WG. Statistical Methods, Ei ghth Edition. Ames (IA): Iowa State Press; 1989. 

# Page 349 of 358

# MPI Research Study Number: 1271-001

18 11. DATA AND SPECIMEN RETENTION

All raw data, documentation, records, protocol, re serve samples, specimens (slides, blocks, and

wet tissues), and the final report generated as a result of this study w ill be retained at MPI

Research, or an approved archive facility contracted by MPI Research, for a period of 1 year

following completion of the study (fin al report issue date). Retent ion of materials after the time

stated above will be subject to a future cont ractual agreement betwee n the Sponsor and MPI

Research.

12. ANIMAL WELFARE

MPI Research is committed to complying with all applicable regulati ons governing the care and

use of laboratory animals. Animal welfare for this study will be in compliance with the U.S.

Department of Agriculture’s (USDA) Animal We lfare Act (9 CFR Parts 1, 2 and 3). The Guide

for the Care and Use of Laboratory Animals, Ins titute of Laboratory Animal Resources, National

Academy Press, Washington, D.C., 1996, will be fo llowed. This facility maintains an Animal

Welfare Assurance statement with National Inst itutes of Health, Office of Laboratory Animal

Welfare.

To ensure compliance:

A. This protocol will be reviewed and a pproved by the Institutional Animal Care and Use

Committee (IACUC) before animal receipt or transfer.

B. The Sponsor, by his or her signature, attests th at the activities specified in this protocol do not

unnecessarily duplicate any pr evious experiment.

C. The Study Director has considered alternativ es to procedures that may cause more than

momentary or slight pain or distress to the animal and has signified that (select one):

__ i.) The relevant supervisory government agency currently gives no alternatives.

X ii.) The following literature searches have been performed to determine whether an

alternative species could be used or anothe r procedure to reduce any pain or distress

was available and none was found.

Date: February 14, 2006

Interval Searched: All years to Present

Search terms: general alternative test ing methods; alterna tive testing methods,

toxicology; general toxicology testing method alternatives

Databases searched: toxnet.nlm.ni h.gov;pubmed.gov;medscape.com;caat.jhsph.edu

Date: February 14, 2006

Interval Searched: All years to Present

Search terms: alternative testing methods rodents; al ternative toxicology methods

rodents; rodents, alternative te sting; testing methods rodents

Databases searched: toxnet.nlm.nih.gov; pubmed.gov; medscape.com; caat.jhsph.ed

__ iii.) This study does not require any procedures that may cause more than slight or

momentary pain or distress to the animal . Note, unknown test articles are presumed

to have the potential to cause more than slight pain or distress. Page 350 of 358

# Page 351 of 358

# Page 352 of 358

# Page 353 of 358

# Page 354 of 358

# Page 355 of 358

# Page 356 of 358

# 1271-001 Appendix S

Deviations Page 357 of 358

# 1271-001 DEVIATIONS

This study was conducted in accordance with the protocol, except for the following

deviations.

Samples for stability analyses of the dosing formulations at the low and high

concentration were collected after 7 and 14 days of refrigeration. However, the

samples were further stored refrigerated and analyzed 2 to 44 days after the samples

were collected.

On Day 2, the postdose observations for tw o control males (animal numbers 311 and

312) were conducted 13 minutes outside the 30 to 90 minute allowable window.

On one occasion, the P.M. cagesid e observation was not documented.

On July 4, 2006, test article administrati on was conducted 11 mi nutes outside the

allowable window of ± 2 hours from the start of dosing on Day 1.

On July 16, 2006, vaginal lavage for estrous cycle evaluation for eight females at

2400 mg/kg/day (animal numbers 389 to 396) was conducted after 10:00 A.M.

The body weight of one male at 150 mg /kg/day (animal number 313) was not

documented at the time of necropsy.

Fetuses from females euthanized in extremis , incomplete deliveries, or died pregnant

were not saved in 10% neutral buffered fo rmalin for possible further examination.

In the opinion of the Study Director, these minor deviations did not a ffect the quality or

integrity of the study. Page 358 of 358