REVISED FINAL REPORT 

 

AN ORAL REPRODUCTION/DEVELOPMENTAL TOXICITY 

SCREENING STUDY OF ANTHRAQUINONE (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 original final report of MPI Research Study Number 

1271-001, titled “An Oral Reproduction/Developmental 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 – % Postimplantation 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 discussion 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 F1 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 Delivery and Litter Data – Added 

% Postimplantation Loss. 

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

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

11. Appendix N. Individual P Natural Delivery 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
1271-001

Page 3 of 358
1271-001

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 Article 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 F1 Litter Data....................................................................................30 

5.6. F1 Pup Evaluations ....................................................................................................30 

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

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

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

Table 

1. Summary of P Clinical Findings - MALE .........................................................................38 

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

3. Summary of P Gestation Clinical Findings .......................................................................44 

4. Summary of P Lactation Clinical Findings........................................................................47 

5. Summary of P Body Weight Values - MALE ...................................................................50 

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

7. Summary of P Gestation Body Weight Values..................................................................54 

8. Summary of P Lactation Body Weight Values..................................................................56 

Page 6 of 358
1271-001 

TABLE OF CONTENTS 

Page 

Table (Cont.) 

9. Summary of P Gestation Body Weight Change Values ....................................................58 

10. Summary of P Lactation Body 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 Weight Values – MALE .................................................................75 

18. Summary of P Organ Weight Values – FEMALE.............................................................77 

19. Summary of P Microscopic 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 F1 Pup Clinical Findings................................................................................92 

25. Summary of F1 Pup Body Weight Values..........................................................................94 

26. Summary of F1 Pup Macroscopic Observations ................................................................96 

Appendix 

A. Vehicle and Test Article Information..............................................................................103 

B. Analytical Report.............................................................................................................106 

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 Organ Weight Values..................................................................................191 

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

L. Individual P Premating/Mating Estrous Cycling Data.....................................................229 

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

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

O. Individual F1 Pup Clinical Findings.................................................................................245 

Page 7 of 358
1271-001 

TABLE OF CONTENTS 

Page 

Appendix (Cont.) 

P. Individual F1 Pup Body Weight, Sex, and Status Data ....................................................275 

Q. Individual F1 Pup Animal Data Record: Pathology........................................................283 

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

S. Deviations ........................................................................................................................357 

Page 8 of 358
1271-001

Page 9 of 358
1271-001 

2. SUMMARY 

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

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

including gonadal function, estrous cycle, mating behavior, conception, 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 the 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 while 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 animals were cohabited, one male and one female 

within the same treatment group for up to 14 days. 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, individual 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 weights were recorded. Selected tissues were 

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

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

Preparation of the dosing suspensions was problematic. Due to the viscosity of the vehicle 

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

difficult to obtain uniform suspensions. Periodic 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 concentration levels achieved. 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 actual 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, for the 150, 600, and 2400 mg/kg/day dose levels. 

Test article-related mortality was seen in female 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 treatment with Anthraquinone. 

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

Anthraquinone, 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 and an erosion/ulcer was 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 died, 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 evident from mating, fertility, 

and fecundity indices but treatment did produce 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 females retained litters through LD 4. In the 

2400 mg/kg/day group, four females delivered, but only one retained 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 mean number of total implantation scars/litter 

was significantly lower in these groups and the mean post-implantation loss was higher 

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

scars, fewer number of pups delivered (live plus stillborn), and increase in post-implantation 

loss) at 600 and 2400 mg/kg/day were considered test article-related. Pup survival to LD 4, 

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

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

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

Mean pup body weights (by sex and sexes combined) 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 notably lower than control. 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 Standard Operating Procedures (SOP) and the 

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

Reproduction/Developmental Toxicity Screening Test, the Organization of Economic 

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

July 1995. The protocol and amendments are presented 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 effects of the test article on male and female 

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

conception, development of the conceptus, and parturition in rats. 

3.2. Species Selection 

The current state of scientific knowledge and 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 knowledge 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 interactions between the various cells, tissues, and 

organs comprising the human body”.2 The rat is the usual rodent model3 used for evaluation 

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

3.3. Justification for Number on Study 

This study was designed to use the fewest number of animals possible, consistent with the 

objective of the study, the scientific needs of the Sponsor, contemporary scientific standards, 

and in consideration of applicable regulatory 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 (receipt 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, composition, 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 provided 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 distilled water, the required amount of powdered 

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

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

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

the contents mixed thoroughly between additions, and the contents were transferred into an 

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

rinse was transferred into the cylinder. Distilled 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 vehicle. The contents of the container were 

thoroughly mixed using a motor-driven propeller. 

Fresh vehicle was prepared for use on study weekly, prior to handling the test article, and 

was stored refrigerated. To dispense the vehicle for dosing, 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 glass 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 article formulations at each concentration, the required amount of 

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

required amount of test article, 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 contents of the beaker were transferred into a 

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

was transferred into the cylinder. 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 evenly dispensed into amber glass containers, 

while stirring, using a syringe. 

Page 14 of 358
1271-001 

Formulations of the test article were prepared for each concentration weekly, and were stored 

refrigerated. On occasion, additional preparations 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 article 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 batch 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 vials. Following completion of the study, any 

remaining samples were properly disposed. 

4.1.2.2. Stability 

Samples (2.0 mL) of the prepared formulations 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 completion 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 the test article. The samples were collected 

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

scintillation vials. Following completion of the study, any remaining samples were properly 

disposed. 

4.1.2.4. Analyses 

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

MPI Research, Inc., using an analytical method 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 used in this study was collected 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 weighed and observed twice daily with respect 

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

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

performed during the acclimation period are not reported, but are maintained 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 suitable for study were weighed prior to treatment 

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

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

via carbon dioxide inhalation and discarded. 

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

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

groups identified in the table below. 

Group Assignments 

Group Number of Animals 

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 ProvantisTM data collection 

system and was implanted with a microchip bearing a unique identification number. 

The individual animal number, implant number, 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 during the course of the study, as documented in the data. 

Animals found dead during the first 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 animals and were assigned a unique animal number. 

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

file. 

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

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

humidity were continuously monitored and recorded. The protocol-designated 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 maintained in the study file. Animals were individually housed in 

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

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

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

individually housed in plastic cages containing wood chip bedding. 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 Nutrition International, Inc.) was available

ad libitum. The lot number from each diet lot used for 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 intervals according to SOP. The results of food and water analyses 

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

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

study. Therefore, no analyses other than 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 exposure 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 administration 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 was performed at approximately the same time 

each day (± 2 hours from Day 1 start of dosing). Individual doses were 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 test article prior to administration. 

4.3. In-life Examinations 

4.3.1. Cageside Observations 

All animals were observed for morbidity, mortality, injury, and the availability of food and 

water twice daily throughout the duration of the study. On occasion, veterinary consultations 

Page 17 of 358
1271-001 

were conducted during the course of the study. All treatments and observations were 

recorded. The medical treatments and observations 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 examination. The observations included, but were 

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

external genitalia, limbs and feet, as well as evaluation of respiration. 

4.3.3. Body Weights and Body Weight Changes 

Individual body weights for the males and females were recorded the day after arrival, prior 

to randomization, at initiation of test article administration, 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 weight 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 initiation of test article administration are not reported, but are 

maintained in the study data. 

4.3.4. Food Consumption 

Food consumption for all rats was measured and recorded weekly prior to pairing. Food 

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

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

unmated females until euthanasia. Food consumption 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 randomly 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 copulation was established by daily inspection for 

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

which positive evidence of copulation was observed 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 cages until scheduled euthanasia. 

Page 18 of 358
1271-001 

After the mating period, the males were individually housed and continued on treatment for 

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

as described in Section 4.4. Postmortem Study Evaluations. 

4.3.7. P Parturition and F1 Litter Observations 

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

parturition. The mated females were allowed to give birth (F1). 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 parameters including litter size, the sex of the pups, number of 

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

pups found dead at birth were externally examined, 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 intraperitoneal injection of sodium pentobarbital 

solution, externally examined and any abnormalities were recorded, and the carcasses were 

discarded. 

4.4. Postmortem Study Evaluations 

4.4.1. Macroscopic 

Necropsy examinations were performed under 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 external abnormalities including masses. The skin 

was reflected from a ventral midline incision and any abnormalities were identified and 

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

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

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

sites (scars) along the uterus were counted and recorded. All designated 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-designated organ weights were recorded for all surviving adult 

animals at the scheduled necropsy and appropriate 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 weight 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 microscopic 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 used 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 below and on the following 

page. 

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 Changes Group Pair-wise Comparisons 

 Gestation Food Consumption Group Pair-wise Comparisons 

 Lactation Body Weights Group Pair-wise Comparisons 

 Lactation Body Weight Changes Group Pair-wise Comparisons 

 Lactation Food Consumption Group Pair-wise Comparisons 

 Postmating Body Weights - Males Group Pair-wise Comparisons 

 Postmating Food Consumption - Males Group Pair-wise Comparisons 

Fertility Indices 

 Estrous Cycle (mean cycle time) Group Pair-wise Comparisons

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

 Gestation Length Group Pair-wise Comparisons 

 Copulatory Interval Group Pair-wise Comparisons 

Page 20 of 358
1271-001 

Endpoint Analysis 

 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 

Pathology

 Male Reproduction Organ Weights Group Pair-wise Comparisons 

 Female Reproduction Organ Weights Group Pair-wise Comparisons 

Litter 

 Number of Implantation scars/dam Group Pair-wise Comparisons

 Litter Size Group Pair-wise Comparisons 

 Viable Pups Group Pair-wise Comparisons 

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

 Stillborn Pups Group Pair-wise Comparisons 

 Stillborn Index Arcsin-Square Root Transformation

 % Postimplantation Loss Arcsin-Square Root Transformation

 Pup Weights Covariate Analysis 

 Pup Survival (Days 0-4) Arcsin-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 test4 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’s5

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

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

t-test6

 with a Bonferroni correction. 

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

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

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

assumption (n>3) were conducted using Welch’s t-test 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-test7

. 

 

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 difference 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 comparisons 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 Followed by Group Pair-wise Comparisons 

Data comprised of percent values were transformed using the arcsin of the square root7

. The 

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

transformed percentage values. 

4.5.3. Fisher’s Exact Test 

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

Fisher’s exact8 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 were 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 testing 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 covariance9

 was 

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

dam) and time period. The covariate was litter size. If the control group did not satisfy the 

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

satisfying the sample size assumption was compared to control using Dunnett’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 reported at the 0.05 and 0.01 significance 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. 4th ed. New Jersey: Prentice Hall; 1999. 9

 Snedecor, GW, Cochran, WG. Statistical Methods, Eighth 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 

ProvantisTM 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: Millennium32 HPLC System (instrument) and 

Millennium32 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 developed 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 applications development. 

Microsoft® Office 2003 Professional: Bundle of integrated productivity tools including word 

and data processing and communications software. 

Contains the utilities Microsoft® Access, Excel, InfoPath, 

Outlook, PowerPoint, Publisher, and Word. 

Microsoft® Office 2000 Professional: Microsoft® Excel 2000 used in conjunction with 

Millennium32 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 Research, Inc., for a period of 1 year following 

completion of the study (final report issue date). Retention of materials after the time stated 

above will be subject to future contractual agreements. 

Page 23 of 358
1271-001 

5. RESULTS AND DISCUSSION 

5.1. Analysis of Dosing Formulations 

The individual results of the dosing formulation analysis for homogeneity, stability, and 

concentration are presented in Appendix B and are summarized below. 

Preparation of the dosing suspensions was problematic. Due to the viscosity of the vehicle 

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

difficult to obtain uniform suspensions. Periodic 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 suspensions was conducted at the low and high 

concentrations prior to study initiation (first test batch) 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 from the second test batch preparation. Mean 

recovery for the 150 mg/kg/day level was again 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 weight/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 % 

Recoverya

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 % 

Recoverya

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 % 

Recoverya

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 formulations 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 article 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 study 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 average 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, for 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) 

% Recoverya

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) 

% Recoverya

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) 

% Recoverya

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 restricted to females at the higher dose levels. In the 600 and 

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

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

were observed in the glandular stomach of several 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 examination or microscopically with erosion/ulcer of 

the stomach. All females in the control and 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 absence of mortality among 

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

5.2.2. Clinical Findings 

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

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

Test article-related clinical findings of discolored urine and decreased activity were noted in 

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

were observed at one or more daily intervals with discolored urine (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 dissimilar observations were noted at clinical 

examination of the treated animals but these were 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 (females – premating/mating), 7 

(females – gestation), and 8 (females – lactation). Body weight change data are summarized 

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

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

Mean body weights of treated males were lower than controls throughout 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 females were also lower than 

control throughout the premating, mating, gestation, and lactation period and in most 

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

significantly lower than control 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 statistically significant, 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. Body 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 Tables 11 (males), 12 (females – premating), 13 

(females – gestation), and 14 (females – lactation) and presented individually in Appendix G. 

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

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

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

(premating period) and was significantly 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 lower 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 presented below and in Appendix H. 

5.3.1. P Macroscopic 

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

macroscopic observations are presented in Appendix K. 

Test article-related macroscopic changes were 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-related 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 changes 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 controls, but the relative increases were caused by 

decreased body weight and were not 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 considered 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 summarized 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 moderate periportal vacuolation of hepatocytes were observed in 

one female at 2400 mg/kg/day. Edema (2 of 7) and 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 treated and control rats, and 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 only statistically significant at 600 mg/kg/day. 

Therefore, with the increase in cycle length, the number of cycles 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/day 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 Anthraquinone 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 interval (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 F1 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 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 females retained litters through LD 4. Two females died pregnant, 

two females died during delivery, four females died during the lactation 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 litter 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 entire 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 stillborn 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. Mean 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 total implantation scars, fewer number of pups delivered (live 

plus stillborn), and increase in post-implantation loss) at 600 and 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 controls and unaffected by treatment with 

Anthraquinone. 

5.6. F1 Pup Evaluations 

5.6.1. Clinical Observations 

F1 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 unrelated to treatment. Several pups 

were noted with skin cold to touch in the 600 mg/kg/day group. These 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 observations were noted for these pups. 

Page 30 of 358
1271-001 

5.6.2. Body Weights 

F1 pup body weight values are summarized in Table 25 and presented individually in 

Appendix P. 

Mean pup body weights (by sex and sexes combined) 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 significantly lower (22-25%) in comparison to 

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

analysis could not be conducted due to the small 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 data 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 

F1 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 considered comparable to controls 

and unaffected by treatment. 

5.6.5. Pup Macroscopic 

Pup macroscopic observations are summarized in Table 26 and presented individually in 

Appendix Q. 

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

necropsy. 

 

Page 31 of 358
1271-001 

6. CONCLUSION 

In this reproduction/developmental toxicity screening study in rats administered 

Anthraquinone orally, neither a paternal, maternal, nor a developmental No-Observed-EffectLevel (NOEL) was achieved at the dose levels evaluated. 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. Considerable mortality was seen in females 

at the 600 and 2400 mg/kg/day levels and ulcer/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 lactation period. Other effects 

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

increased in stillborn pups, and increased post-implantation loss. Pup survival was poor in 

the 600 and 2400 mg/kg/day groups with only two and 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 parameters in generation 

studies. They are usually classed as primary

(effects related to sex organs or reproductive 

capacity of adults), or secondary (changes that 

may be dependent upon or related 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 Clinical Findings - MALE 

Page 38 of 358
MPI Research Study Number 1271-001 

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

Table 1 Summary of P Clinical Findings+ - MALE 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Number of Animals Alive at Start of Interval 12 12 12 Behavior/Activity 

Activity decreased 0/0 3/1 0/0 Salivation 0/0 0/0 1/1 

Excretion 

Urine discolored, Brown 0/0 0/0 5/2 Urine discolored, Orange 2/1 40/12 51/12 Urine discolored, Red 0/0 66/12 213/12 

External Appearance 

Carriage low 0/0 2/1 0/0 Malocclusion 0/0 0/0 8/1 Material around eyes, Black, Eye/left 0/0 0/0 4/1 Material around eyes, Black, Eye/right 0/0 0/0 4/1 

Material around eyes, Red, Eye/left 0/0 0/0 4/1 Material around eyes, Red, Eye/right 0/0 0/0 4/1 Material around nose, Black 0/0 0/0 0/0 Material around nose, Red 0/0 2/1 0/0 Posture hunched 0/0 9/1 0/0 

Swelling, Anogenital region 0/0 0/0 0/0 

Pelage/Skin 

Abrasion(s), Cervical region 0/0 0/0 21/1 Abrasion(s), Lip 0/0 0/0 0/0 Hair discolored, Brown, Anogenital region 0/0 0/0 0/0 Hair discolored, Brown, Forelimb/left 0/0 15/1 0/0 Hair discolored, Brown, Forelimb/right 0/0 15/1 0/0 Hair discolored, Brown, Ventral surface 0/0 0/0 0/0 Hair discolored, Yellow, Anogenital region 1/1 0/0 0/0 
MPI Research Study Number 1271-001 

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

Table 1 Summary of P Clinical Findings+ - MALE 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Pelage/Skin 

Hair discolored, Yellow, Ventral surface 0/0 0/0 0/0 Hair sparse, Cervical region 0/0 0/0 11/1 Hair sparse, Face 0/0 11/1 0/0 Hair sparse, Forefoot/left 25/1 11/1 0/0 Hair sparse, Forefoot/right 25/1 11/1 0/0 Hair sparse, Forelimb/left 0/0 36/2 0/0 Hair sparse, Forelimb/right 0/0 36/2 21/2 Hair sparse, Ventral surface 0/0 9/1 0/0 Piloerection 0/0 12/1 0/0 Scabbed area, Thoracic region 0/0 0/0 7/1 

Respiration 

Breathing difficult 0/0 3/1 0/0 

Rales 0/0 0/0 0/0 
1271-001 

Table 2 

Summary of P Premating/Mating Clinical Findings – FEMALE 

Page 41 of 358
MPI Research Study Number 1271-001 

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

Table 2 Summary of P Premating/Mating Clinical Findings+ - FEMALE 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Number of Animals Alive at Start of Interval 12 12 12 Behavior/Activity 

Activity decreased 0/0 0/0 8/3 Hypersensitive to touch 0/0 0/0 3/1 

Excretion 

Feces discolored, Yellow 0/0 0/0 0/0 Feces few/absent 0/0 5/2 2/2 Feces watery 0/0 0/0 0/0 Urine discolored, Brown 0/0 3/1 1/1 Urine discolored, Orange 1/1 2/2 7/3 Urine discolored, Red 0/0 4/1 106/12 

External Appearance 

Lacrimation, Eye/right 1/1 0/0 0/0 

Malocclusion 16/1 0/0 0/0 Material around eyes, Red, Eye/right 16/1 0/0 0/0 

Material around mouth, Red 0/0 0/0 2/1 Material around nose, Red 2/1 0/0 0/0 Posture hunched 0/0 5/2 9/3 Swelling, Dorsal surface 0/0 0/0 11/1 Thin 0/0 0/0 0/0 

Pelage/Skin 

Hair discolored, Brown, Anogenital region 0/0 0/0 2/2 Hair discolored, Yellow, Anogenital region 0/0 0/0 2/2 Hair discolored, Yellow, Ventral surface 0/0 8/1 0/0 Hair sparse, Abdominal region 0/0 19/1 22/1 Hair sparse, Hind limb/left 0/0 0/0 0/0 Hair sparse, Hind limb/right 0/0 0/0 0/0 
MPI Research Study Number 1271-001 

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

Table 2 Summary of P Premating/Mating Clinical Findings+ - FEMALE 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Pelage/Skin 

Hair sparse, Inguinal region/right 0/0 0/0 0/0 

Hair sparse, Lumbar region 0/0 0/0 0/0 Hair sparse, Ventral surface 0/0 12/1 0/0 Piloerection 0/0 0/0 7/3 Scabbed area, Dorsal surface 0/0 0/0 3/1 Skin discolored, Pale, Entire body 0/0 0/0 0/0 Unkempt appearance 0/0 0/0 0/0 

Respiration 

Breathing audible 1/1 3/1 5/2 Breathing rapid 0/0 0/0 2/1 Rales 0/0 0/0 3/2 
1271-001 

Table 3 

Summary of P Gestation Clinical Findings 

Page 44 of 358
MPI Research Study Number 1271-001 

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

Table 3 Summary of P Gestation Clinical Findings+ 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Number of Animals Alive at Start of Interval 11 11 10 Behavior/Activity 

Activity decreased 0/0 0/0 10/4 

Excretion 

Feces discolored, Yellow 0/0 0/0 1/1 Feces soft 0/0 0/0 0/0 Feces watery 0/0 0/0 2/1 Material in pan/bedding, Red 0/0 0/0 0/0 Urine discolored, Orange 1/1 8/7 36/10 Urine discolored, Red 0/0 48/11 66/10 

External Appearance 

Carriage high 0/0 0/0 12/2 Discharge, Clear, Anus 0/0 0/0 0/0 Discharge, Red, Vulva 0/0 0/0 2/2 Malocclusion 27/1 0/0 0/0 Material around eyes, Red, Eye/right 14/1 0/0 0/0 

Material around mouth, Red 0/0 0/0 5/1 Material around nose, Black 0/0 0/0 0/0 Material around nose, Red 0/0 0/0 7/2 Posture hunched 0/0 3/1 0/0 Thin 0/0 0/0 16/2 

Eye/Ocular 

Eye discolored, Pale, Eye/left 0/0 0/0 1/1 Eye discolored, Pale, Eye/right 0/0 0/0 1/1 Eyelid partially/completely closed, Eye/left 0/0 0/0 1/1 Eyelid partially/completely closed, Eye/right 0/0 0/0 1/1 
MPI Research Study Number 1271-001 

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

Table 3 Summary of P Gestation Clinical Findings+ 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Pelage/Skin 

Hair discolored, Brown, Anogenital region 0/0 6/1 18/2 Hair discolored, Red, Abdominal region 0/0 0/0 2/1 Hair discolored, Red, Anogenital region 0/0 0/0 19/5 Hair discolored, Red, Ventral surface 0/0 10/1 0/0 Hair discolored, Yellow, Anogenital region 0/0 0/0 1/1 Hair discolored, Yellow, Ventral surface 0/0 10/1 0/0 Hair sparse, Abdominal region 0/0 28/1 37/2 Hair sparse, Anogenital region 0/0 14/1 12/1 Hair sparse, Cervical region 0/0 0/0 12/1 Hair sparse, Forelimb/left 0/0 27/2 11/1 Hair sparse, Forelimb/right 0/0 27/2 11/1 Hair sparse, Hind limb/left 0/0 0/0 0/0 Hair sparse, Hind limb/right 0/0 0/0 0/0 Hair sparse, Lumbar region 0/0 0/0 0/0 Hair sparse, Ventral surface 0/0 27/1 0/0 Piloerection 0/0 0/0 20/5 Scabbed area, Dorsal surface 0/0 0/0 10/1 Skin cold to touch 0/0 0/0 12/4 Skin discolored, Blue, Abdominal region 0/0 0/0 0/0 Skin discolored, Pale, Entire body 0/0 0/0 3/2 

Respiration 

Breathing shallow 0/0 0/0 1/1 

Breathing slow 0/0 0/0 1/1 Rales 0/0 1/1 0/0 
1271-001 

Table 4 

Summary of P Lactation Clinical Findings 

Page 47 of 358
MPI Research Study Number 1271-001 

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

Table 4 Summary of P Lactation Clinical Findings+ 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Number of Animals Alive at Start of Interval 11 10 9 Behavior/Activity 

Activity decreased 0/0 0/0 7/3 Tremors 0/0 0/0 2/1 

Excretion 

Feces watery 0/0 0/0 1/1 

External Appearance 

Carriage high 0/0 0/0 11/3 Discharge, Red, Vulva 0/0 0/0 2/2 Malocclusion 5/1 0/0 0/0 Material around eyes, Red, Eye/right 3/1 0/0 0/0 Material around mouth, Red 0/0 0/0 2/1 Material around nose, Red 0/0 0/0 4/2 Thin 0/0 0/0 12/3 

Eye/Ocular 

Eye discolored, Pale, Eye/left 0/0 0/0 1/1 Eye discolored, Pale, Eye/right 0/0 0/0 1/1 

Pelage/Skin 

Hair discolored, Red, Abdominal region 0/0 0/0 2/1 Hair discolored, Red, Anogenital region 0/0 0/0 4/3 Hair sparse, Abdominal region 0/0 5/1 8/2 Hair sparse, Anogenital region 0/0 5/1 4/1 Hair sparse, Cervical region 0/0 0/0 4/1 Hair sparse, Forelimb/left 0/0 5/1 5/1 Hair sparse, Forelimb/right 0/0 5/1 5/1 Hair sparse, Hind limb/left 0/0 0/0 0/0 
MPI Research Study Number 1271-001 

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

Table 4 Summary of P Lactation Clinical Findings+ 

 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+Number of times observed/Total number of animals affected 

Pelage/Skin 

Hair sparse, Hind limb/right 0/0 0/0 0/0 Hair sparse, Ventral surface 0/0 4/1 0/0 Piloerection 0/0 0/0 8/3 Skin cold to touch 0/0 0/0 11/5 Skin discolored, Pale, Entire body 0/0 0/0 2/2 

Respiration 

Breathing shallow 0/0 0/0 1/1 

Breathing slow 0/0 0/0 0/0 
1271-001 

Table 5 

Summary of P Body Weight Values - MALE 

Page 50 of 358
MPI Research Study Number 1271-001 

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

Table 5 Summary of P Body Weight Values - MALE  Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Week) Mean SD N Mean SD N Mean SD N Body Weight Values g 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

Premating 1 260.3 9.33 12 265.1 13.55 12 263.1 16.12 12 

 2 305.2 14.68 12 298.8 24.58 12 293.5 17.64 12 Pairing 3 348.9 21.20 12 337.1 24.20 12 326.8a 18.59 12  4 378.2 26.60 12 366.9 22.90 12 350.6a 22.62 12 Postpairing 5 408.8 33.95 12 393.5 20.33 12 380.3a 23.85 12  6 444.3 38.93 12 426.2 21.30 11 410.2a 20.87 12  7 469.3 41.65 12 440.0 23.08 10 415.9b 24.37 11 

aSignificantly different from control; (p<0.05) bSignificantly different from control; (p<0.01) 
1271-001 

Table 6 

Summary of P Premating/Mating Body Weight Values - FEMALE 

Page 52 of 358
MPI Research Study Number 1271-001 

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

Table 6 Summary of Premating Body Weight Values - FEMALE 

 Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Week) Mean SD N Mean SD N Mean SD N Body Weight Values g 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 1 195.4 8.39 12 193.8 5.86 12 195.8 8.90 12 

 2 214.3 12.32 12 205.8 10.09 12 190.9b 16.33 12  3 229.5 14.37 12 212.2b 8.72 10 206.9b 14.28 11 

bSignificantly different from control; (p<0.01) 
1271-001 

Table 7 

Summary of P Gestation Body Weight Values 

Page 54 of 358
MPI Research Study Number 1271-001 

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

Table 7 Summary of P Gestation Body Weight Values  Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Day) Mean SD N Mean SD N Mean SD N Body Weight Values g 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 0 248.9 16.66 11 218.2b 15.79 10 209.7b 24.17 10 

 7 288.8 20.75 11 252.9b 12.47 10 233.9b 24.71 10  14 324.2 24.00 11 288.2b 15.84 10 263.1b 28.59 10  20 405.0 32.59 11 347.5b 25.23 10 303.3b 49.05 10 

aSignificantly different from control; (p<0.05) bSignificantly different from control; (p<0.01) 
1271-001 

Table 8 

Summary of P Lactation Body Weight Values 

Page 56 of 358
MPI Research Study Number 1271-001 

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

Table 8 Summary of P Lactation Body Weight Values 

 Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Day) Mean SD N Mean SD N Mean SD N Body Weight Values g 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 0 300.9 25.49 11 252.1b 21.75 10 201.8b 27.34 9 

 4 314.5 22.52 11 269.0b 39.14 10 203.5b 28.64 4 

bSignificantly different from control; (p<0.01) 
1271-001 

Table 9 

Summary of P Gestation Body Weight Change Values 

Page 58 of 358
MPI Research Study Number 1271-001 

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

Table 9 Summary of P Gestation Body Weight Change Values 

 Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Day) Mean SD N Mean SD N Mean SD N Body Weight Change Values g 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 0-7 39.9 8.07 11 34.7 6.18 10 24.2b 9.59 10 

 7-14 35.4 7.46 11 35.3 7.12 10 29.2 11.52 10  14-20 80.8 11.69 11 59.3 13.54 10 40.2b 25.42 10  0-20 156.1 21.64 11 129.3 14.66 10 93.6b 33.12 10 

bSignificantly different from control; (p<0.01) 
1271-001 

Table 10 

Summary of P Lactation Body Weight Change Values 

Page 60 of 358
MPI Research Study Number 1271-001 

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

Table 10 Summary of P Lactation Body Weight Change Values 

 Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Day) Mean SD N Mean SD N Mean SD N Body Weight Change Values g 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 0-4 13.5 12.76 11 16.9 27.65 10 7.8 20.42 4 
1271-001 

Table 11 

Summary of P Food Consumption Values – MALE 

Page 62 of 358
MPI Research Study Number 1271-001 

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

Table 11 Summary of P Food Consumption Values - MALE  Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Week) Mean SD N Mean SD N Mean SD N Food Consumption Values g/animal/day 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

Premating 1-2 26.5 2.11 12 22.4b 3.52 12 22.5b 1.53 12 

 2-3 27.5 2.44 12 25.4 2.72 12 25.3 1.96 11 Postmating 5-6 28.5 3.44 12 27.0 1.45 11 25.8a 1.63 12  6-7 28.1 3.16 12 25.3a 1.46 11 24.9a 2.97 12 

aSignificantly different from control; (p<0.05) bSignificantly different from control; (p<0.01) 

NA - Not Applicable/Not Available 
1271-001 

Table 12 

Summary of P Premating Food Consumption Values - FEMALE 

Page 64 of 358
MPI Research Study Number 1271-001 

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

Table 12 Summary of P Premating Food Consumption Values - FEMALE  Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Week) Mean SD N Mean SD N Mean SD N Food Consumption Values g/animal/day 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 1-2 17.9 2.17 12 14.4b 1.69 12 10.8b 1.69 12 

 2-3 19.3 2.17 12 17.2a 1.96 12 16.7b 2.10 11 

aSignificantly different from control; (p<0.05) bSignificantly different from control; (p<0.01) 
1271-001 

Table 13 

Summary of P Gestation Food Consumption Values 

Page 66 of 358
MPI Research Study Number 1271-001 

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

Table 13 Summary of P Gestation Food Consumption Values  Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Day) Mean SD N Mean SD N Mean SD N Food Consumption Values g/animal/day 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 0-7 24.9 2.83 11 20.4b 1.35 10 19.1b 2.59 10 

 7-14 25.6 3.36 11 21.6b 2.65 10 19.4b 3.02 10  14-20 28.0 2.43 11 23.3a 3.66 10 20.4b 4.43 10  0-20 26.1 2.74 11 21.7b 1.94 10 19.6b 2.96 10 

aSignificantly different from control; (p<0.05) bSignificantly different from control; (p<0.01) 
1271-001 

Table 14 

Summary of P Lactation Food Consumption Values 

Page 68 of 358
MPI Research Study Number 1271-001 

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

Table 14 Summary of P Lactation Food Consumption Values 

 Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

 Interval Endpoint (Day) Mean SD N Mean SD N Mean SD N Food Consumption Values g/animal/day 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

 0-4 37.1 4.70 11 32.4 6.87 9 12.3b 2.39 4 

bSignificantly different from control; (p<0.01) 
1271-001 

Table 15 

Summary of P Macroscopic Observations – MALE 

Page 70 of 358
MPI Research Study Number 1271-001 

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

Summary of P Macroscopic Observations - MALE 

Table 15 Terminal 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/da

Tissue 

Observation Severity DOS SNC DOS SNC DOS SNC 

Number of Animals Examined 0 12 1 11 0 12 

DOS - Died or euthanized on study 

SNC - Scheduled necropsy 

0 0 0 0 

all tissues 

within normal limits 0 12 0 10 11 

cavity, thoracic 

adhesion - mild 0 0 0 1 0 

skin 

abrasion - mild 0 0 0 0 1 

thyroid gland 

discoloration, red - mild 0 0 1 0 0 
1271-001 

Table 16 

Summary of P Macroscopic Observations – FEMALE 

Page 72 of 358
MPI Research Study Number 1271-001 

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

Summary of P Macroscopic Observations - FEMALE 

Table 16 Terminal 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/da

Tissue 

Observation Severity DOS SNC DOS SNC DOS SNC 

Number of Animals Examined 0 12 0 12 8 4 

DOS - Died or euthanized on study 

SNC - Scheduled necropsy 

2 0 1 0 1 1 2 1 1 0 0 

all tissues 

within normal limits 0 12 0 11 4 

animal/whole body 

body fat depleted - mild 0 0 0 0 0 

kidneys 

discoloration - mild 0 0 0 0 0 

liver 

discoloration, pink - mild 0 0 0 0 0 lobulation, accentuated - moderate 0 0 0 0 0 

stomach, glandular 

focus/foci, black - mild 0 0 0 0 0 focus/foci, red 0 0 0 0 0 

- minimal 0 0 0 0 0 

- moderate 0 0 0 0 0 

stomach, nonglandular 

focus/foci, red - mild 0 0 0 1 0 

stomach, pylorus 

erosion/ulcer - moderate 0 0 0 0 0 
MPI Research Study Number 1271-001 

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

Summary of P Macroscopic Observations - FEMALE 

Table 16 Terminal 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/da

Tissue 

Observation Severity DOS SNC DOS SNC DOS SNC 

Number of Animals Examined 0 12 0 12 8 4 

DOS - Died or euthanized on study 

SNC - Scheduled necropsy 

2 1 1 2 1 1 1 

thymus gland 

discoloration, red 0 0 0 0 0 

- mild 0 0 0 0 0 

- moderate 0 0 0 0 0 

small 0 0 0 0 0 

- mild 0 0 0 0 0 

- moderate 0 0 0 0 0 

thyroid gland 

pale - mild 0 0 0 0 0 
1271-001 

Table 17 

Summary of P Organ Weight Values – MALE 

Page 75 of 358
MPI Research Study Number 1271-001 

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

Summary of P Organ Weight Values - MALE 

Table 17 Terminal 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint Mean SD N Mean SD N Mean SD N M

Body weight g 474 43 12 449 21 10 426b 22 12 430

Epididymides g 1.221 0.100 12 1.196 0.126 11 1.149 0.068 12 1

Epididymides/BWt % 0.2589 0.0254 12 0.2688 0.0253 10 0.2703 0.0214 12 0Prostate gl g 0.728 0.227 12 0.620 0.115 11 0.588 0.162 12 0Prostate gl/BWt % 0.1541 0.0478 12 0.1415 0.0169 10 0.1380 0.0381 12 0Sem. ves. g 1.801 0.208 12 1.852 0.224 11 1.881 0.236 12 1Sem. ves./BWt % 0.3824 0.0534 12 0.4129 0.0572 10 0.4419 0.0557 12 0Testes g 3.315 0.157 11 3.494 0.217 11 3.527 0.299 12 3Testes/BWt % 0.7069 0.0732 11 0.7837 0.0487 10 0.8314b 0.1027 12 0

N - Number of measures used to calculate mean 

SD - Standard Deviation 

aSignificantly different from control; (p<0.05) bSignificantly different from control; (p<0.01) 
1271-001 

Table 18 

Summary of P Organ Weight Values – FEMALE 

Page 77 of 358
MPI Research Study Number 1271-001 

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

Summary of P Organ Weight Values - FEMALE 

Table 18 Terminal 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint Mean SD N Mean SD N Mean SD N M

Body weight g 302 21 11 261b 39 10 200b 28 4 222

Ovaries g 0.137 0.027 11 0.123 0.017 10 0.117 0.019 4 0

Ovaries/BWt % 0.0451 0.0070 11 0.0479 0.0071 10 0.0584a 0.0048 4 0Uterus w/ cervix g 0.806 0.140 11 0.678 0.133 10 0.564a 0.147 4 0Uterus w/ cervix/BWt % 0.2664 0.0426 11 0.2616 0.0453 10 0.2792 0.0383 4 0

N - Number of measures used to calculate mean 

SD - Standard Deviation 

aSignificantly different from control; (p<0.05) bSignificantly different from control; (p<0.01) 
1271-001 

Table 19 

Summary of P Microscopic Observations – MALE 

Page 79 of 358
MPI Research Study Number 1271-001 

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

Summary of P Microscopic Observations - MALE 

Table 19 Terminal 

0 mg/kg/day 2400 mg/kg/day

Tissue 

Observation Severity DOS SNC DOS SNC 

Number of Animals Examined 0 12 0 12 

DOS - Died or euthanized on study 

SNC - Scheduled necropsy 

( ) - Number observed 

epididymides (0) (12) (0) (12)

within normal limits 0 12 0 12

prostate gland (0) (12) (0) (12)

infiltration, lymphocytic - minimal 0 4 0 4within normal limits 0 8 0 8

seminal vesicles (0) (12) (0) (12)

within normal limits 0 12 0 12

testes (0) (12) (0) (12)

within normal limits 0 12 0 12
1271-001 

Table 20 

Summary of P Microscopic Observations – FEMALE 

Page 81 of 358
MPI Research Study Number 1271-001 

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

Summary of P Microscopic Observations - FEMALE 

Table 20 Terminal 

0 mg/kg/day 2400 mg/kg/day

Tissue 

Observation Severity DOS SNC DOS SNC 

Number of Animals Examined 0 12 7 5 

DOS - Died or euthanized on study 

SNC - Scheduled necropsy 

( ) - Number observed 

liver (0) (0) (0) (1)

infiltration, mononuclear cell - minimal 0 0 0 1necrosis, focal - minimal 0 0 0 1vacuolation, periportal - moderate 0 0 0 1

ovaries (0) (12) (7) (5)

within normal limits 0 12 7 5

stomach, glandular (0) (0) (5) (0)

edema 0 0 2 0

- minimal 0 0 1 0- mild 0 0 1 0

erosion/ulcer - mild 0 0 3 0within normal limits 0 0 2 0

uterus (0) (12) (7) (5)

within normal limits 0 12 7 5
1271-001 

Table 21 

Summary of Premating Estrous Cycling 

Page 83 of 358
MPI Research Study Number 1271-001 

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

Table 21 Summary of P Premating Estrous Cycling  0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint Mean SD N Mean SD N Mean SD N 

N - Number of measures used to calculate mean aSignificantly different fr

SD - Standard Deviation bSignificantly different fr

No. - Number

Mean Cycle Length (Days) 4.4 0.44 11 4.7 1.16 12 5.6a 1.43 10 

No. of Cycles (Count) 3.6 0.50 11 2.5b 1.24 12 2.2b 0.79 10 
1271-001 

Table 22 

Summary of Reproductive and Fertility Parameters 

Page 85 of 358
MPI Research Study Number 1271-001 

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

Table 22 Summary of P Reproductive and Fertility Parameters  0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint 

No. - Number 

No. Females on Study 12 12 12 

No. Females Paired 12 12 12 No. Females Mated 11 11 12 No. Pregnant 11 10 12 

Female Mating Index 91.7 91.7 100.0 Female Fertility Index 91.7 83.3 100.0 Female Fecundity Index 100.0 90.9 100.0 No. Males on Study 12 12 12 

No. Males Paired 12 12 12 

No. Males Mated 11 11 12 No. Males Impregnating a Female 11 10 12 

Male Mating Index 91.7 91.7 100.0 Male Fertility Index 91.7 83.3 100.0 

Male Fecundity Index 100.0 90.9 100.0 
MPI Research Study Number 1271-001 

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

Table 22 Summary of P Reproductive and Fertility Parameters  0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint 

N - Number of measures used to calculate mean 

SD - Standard Deviation 

Females with Confirmed Mating Day 11 11 10 Copulatory Interval (Days) 

Mean 3.0 3.0 3.1 

SD 1.18 2.32 1.54 

N 11 11 8 
1271-001 

Table 23 

Summary of P Natural Delivery and Litter Data 

Page 88 of 358
MPI Research Study Number 1271-001 

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

Table 23 Summary of P Natural Delivery and Litter Data 

____________________________________________________________________________________________________________

SD - Standard Deviation 1Not statistically analyzed aSignificantly different froN - Number of measures used to calculate mean bSignificantly different froNo. - Number 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint No. Females on Study N 12 12 12 No. Females Pregnant N 11 10 12 

Female Fertility Index % 91.7 83.3 100.0 

Females Delivering Litters N 11 10 8 

1

% 100.0 100.0 66.7 

With Stillborn Pups N 0 1 3 

1

% 0.0 10.0 37.5 

With All Stillborn N 0 0 1 

1

% 0.0 0.0 12.5 

Mean 22.1 22.6 23.1b 

SD 0.30 0.52 0.69 

Gestation Length (Days) 

N 11 10 7 

Mean 14.6 12.7 10.6a 

SD 2.01 1.64 4.87 

No. of Pups at Day 0 

(Total Pups Born/Litter) 

N 11 10 8 

Mean 14.6 12.6 8.1a 

SD 2.01 1.65 5.79 

Liveborn/Litter 

N 11 10 8 
MPI Research Study Number 1271-001 

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

Table 23 Summary of P Natural Delivery and Litter Data 

____________________________________________________________________________________________________________

SD - Standard Deviation aSignificantly different froN - Number of measures used to calculate mean No. - Number 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint 

Mean 0.0 0.1 2.5 

SD 0.00 0.32 4.60 

No. of Pups at Day 0 cont. 

Stillborn/Litter 

N 11 10 8 

Gestation Index N 11 10 7 

% 100.0 100.0 87.5 

Mean %/Litter 0.00 0.77 19.97 

SD 0.000 2.433 36.017 

Stillborn Index 

N 11 10 8 

Mean 15.5 13.9 13.0a 

SD 1.44 1.20 2.88 

Total Implantation Scars/Litter 

N 11 10 8 

Mean %/Litter 6.07 8.79 20.48 

SD 6.851 6.849 29.667 

Post-Implantation Loss 

N 11 10 8 
MPI Research Study Number 1271-001 

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

Table 23 Summary of P Natural Delivery and Litter Data 

____________________________________________________________________________________________________________

SD - Standard Deviation NA - Not Applicable/Not Available bSignificantly different froN - Number of measures used to calculate mean No. - Number 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Endpoint 

Mean 14.2 12.4 5.0 

SD 1.83 1.94 5.66 

No. Live Pups/Litter 

Day 4 

N 11 9 2 

Mean %/Litter 54.17 45.01 43.65 

SD 17.161 13.470 28.907 

Sex Ratio (% Males per Animal) 

Pups Day 0 

N 11 10 5 

Mean %/Litter 54.72 45.50 11.11 

SD 17.698 14.645 15.713 

Pups Day 4 

N 11 9 2 

Mean %/Litter 97.19 89.00 16.54b 

SD 5.818 31.429 32.850 

Pup Survival Indices 

Viability Index 

N 11 10 5 
1271-001 

Table 24 

Summary of F1 Pup Clinical Findings 

Page 92 of 358
MPI Research Study Number 1271-001 

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

Summary of F1 Pup Clinical Findings+ 

Table 24  0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Observation 

+ - Number of times observed/Total number of animals affected

Number of Animals Alive at Start of Interval 160 126 59 

Animals with No Abnormalities Detected 160 123 25 

Behavior/Activity  Activity decreased 0/0 0/0 1/1 External Appearance  Limb function impaired, Hind limb/left 0/0 2/2 0/0  Limb function impaired, Hind limb/right 0/0 2/2 0/0  Thin 0/0 0/0 1/1 Pelage/Skin  Abrasion(s), Cranial region 0/0 1/1 0/0  Skin cold to touch 0/0 1/1 33/33  Skin discolored, Pale, Entire body 0/0 1/1 1/1 
1271-001 

Table 25 

Summary of F1 Pup Body Weight Values 

Page 94 of 358
MPI Research Study Number 1271-001 

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

Table 25 Summary of F1 Pup Body Weight Values, g 

Study 0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Interval Sex 

SD - Standard Deviation ( ) - Least square mean b Significantly different 

N - Number of measures used to calculate mean 

Day 0  Males Mean 6.75 (6.82) 6.60 (6.57) 5.06 (4.96)b 

 SD 0.466 0.576 0.788  N 11 10 5  Females Mean 6.45 (6.60) 6.32 (6.30) 5.02 (4.79)b  SD 0.403 0.574 1.017  N 11 10 6  Males + Females Mean 6.60 (6.75) 6.46 (6.43) 5.17 (4.95)b  SD 0.443 0.551 0.988  N 11 10 6 Day 4  Males Mean 9.74 (9.76) 8.75 (8.72) 4.80 (4.81)  SD 0.803 1.242 NA  N 11 9 1 

 Females Mean 9.29 (9.29) 8.53 (8.53) 3.93 (3.93) 

 SD 0.947 1.101 0.182  N 11 9 2 

 Males + Females Mean 9.49 (9.50) 8.63 (8.63) 4.01 (4.01) 

 SD 0.842 1.132 0.299  N 11 9 2 
1271-001 

Table 26 

Summary of F1 Pup Macroscopic Observations 

Page 96 of 358
MPI Research Study Number 1271-001 

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

Summary F1 Pup Macroscopic Observations – MALE 

Table 26 Stillborn Pups 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Tissue

Observation Severity 

Number of Animals Examined 0 0 10 external appearance

within normal limits 0 0 10 
MPI Research Study Number 1271-001 

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

Summary F1 Pup Macroscopic Observations – FEMALE 

Table 26 Stillborn Pups 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Tissue

Observation Severity 

Number of Animals Examined 0 1 15 external appearance

within normal limits 0 1 15 
MPI Research Study Number 1271-001 

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

Summary F1 Pup Macroscopic Observations – MALE 

Table 26 Died on Study Pups 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Tissue

Observation Severity 

Number of Animals Examined 2 6 23 external appearance

within normal limits 2 6 23 
MPI Research Study Number 1271-001 

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

Summary F1 Pup Macroscopic Observations – FEMALE 

Table 26 Died on Study Pups 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Tissue

Observation Severity 

Number of Animals Examined 2 8 21 external appearance

within normal limits 2 8 21 
MPI Research Study Number 1271-001 

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

Summary F1 Pup Macroscopic Observations – MALE 

Table 26 Day 4 Pups 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Tissue

Observation Severity 

Number of Animals Examined 87 51 2 external appearance

within normal limits 87 51 2 
MPI Research Study Number 1271-001 

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

Summary F1 Pup Macroscopic Observations – FEMALE 

Table 26 Day 4 Pups 

0 mg/kg/day 150 mg/kg/day 600 mg/kg/day 

Tissue

Observation Severity 

Number of Animals Examined 69 61 8 external appearance

within normal limits 69 61 8 
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 Characteristics: 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
Page 108 of 358
Page 109 of 358
Page 110 of 358
Page 111 of 358
Page 112 of 358
Page 113 of 358
Page 114 of 358
Page 115 of 358
Page 116 of 358
1271-001 

Appendix B 

Section 2 

Analysis of Dosing Formulations 

Page 117 of 358
MPI Research Study Number 1271-001 

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

Homogeneity Analysis of Dosing Formulations

RSD - Relative Standard Deviation 

1 These data failed to meet acceptance criterion for RSD (≤10%). 2 These data failed to meet concentration acceptance criterion for recovery (±15%). 

Dose Level 

(mg/kg/day) Sample Identification 

Nominal 

Concentration 

(mg/mL) 

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration 

(mg/mL) 

Average 

Dose Level 

(mg/kg/day) %Recovery 

1501,2 Test Batch Top Replicate 1 7.5 4.5637 4.1661 83.3220 60.8 

Test Batch Top Replicate 2 2.5457 33.9 

Test Batch Middle Replicate 1 5.5896 74.5 Test Batch Middle Replicate 2 4.8385 64.5 

Test Batch Bottom Replicate 1 3.2569 43.4 Test Batch Bottom Replicate 2 4.2024 56.0 

24002 Test Batch Top Replicate 1 120.00 79.1361 91.5363 1830.7260 65.9 

Test Batch Top Replicate 2 91.1733 76.0 

Test Batch Middle Replicate 1 98.4989 82.1 Test Batch Middle Replicate 2 92.3053 76.9 

Test Batch Bottom Replicate 1 92.8235 77.4 Test Batch Bottom Replicate 2 95.2806 79.4 
MPI Research Study Number 1271-001 

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

Homogeneity Analysis of Dosing Formulations

RSD - Relative Standard Deviation 

1 These data failed to meet acceptance criterion for RSD (≤10%). 2 These data failed to meet concentration acceptance criterion for recovery (±15%). 

Dose Level 

(mg/kg/day) Sample Identification 

Nominal 

Concentration 

(mg/mL) 

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration 

(mg/mL) 

Average 

Dose Level 

(mg/kg/day) %Recovery 

1501,2 6/26/06 Top Replicate 1 7.5 3.4783 3.7765 75.5300 46.4 

6/26/06 Top Replicate 2 3.3648 44.9 

6/26/06 Middle Replicate 1 4.7963 64.0 6/26/06 Middle Replicate 2 4.0172 53.6 

6/26/06 Bottom Replicate 1 2.6793 35.7 6/26/06 Bottom Replicate 2 4.3232 57.6 

2400 6/26/06 Top Replicate 1 120.00 113.9178 104.9651 2099.3020 94.9 

6/26/06 Top Replicate 2 116.6933 97.2 

6/26/06 Middle Replicate 1 106.3067 88.6 

6/26/06 Middle Replicate 2 99.6576 83.0 

6/26/06 Bottom Replicate 1 94.4905 78.7 6/26/06 Bottom Replicate 2 98.7248 82.3 
MPI Research Study Number 1271-001 

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

Week 3 Homogeneity Analysis of Dosing Formulations

RSD - Relative Standard Deviation 

1 These data failed to meet acceptance criterion for RSD (≤10%). 2 These data failed to meet concentration acceptance criterion for recovery (±15%). 

Dose Level 

(mg/kg/day) Sample Identification 

Nominal 

Concentration 

(mg/mL) 

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration 

(mg/mL) 

Average 

Dose Level 

(mg/kg/day) %Recovery 

1501,2 Top Replicate 1 7.5 5.4518 6.2297 124.5940 72.7 

Top Replicate 2 7.4521 99.4 

Middle Replicate 1 6.1169 81.6 Middle Replicate 2 6.6424 88.6 

Bottom Replicate 1 5.5311 73.7 Bottom Replicate 2 6.1839 82.5 

24001 Top Replicate 1 120.00 114.3310 109.6368 2192.7360 95.3 

Top Replicate 2 118.4224 98.7 

Middle Replicate 1 121.3300 101.1 Middle Replicate 2 79.8885 66.6 

Bottom Replicate 1 112.4813 93.7 Bottom Replicate 2 111.3679 92.8 
MPI Research Study Number 1271-001 

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

Stability Analysis of Dosing Formulations 

RSD - Relative Standard Deviation 

1 These data failed to meet acceptance criterion for recovery (±10%). 2 Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from 

homogeneity analysis of dosing formulations. 3 Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from homogeneity analysis of dosing formulations. 

Stability 

Interval 

(Day) 

Dose Level 

(mg/kg/day) Sample Identification 

Initial Calculated 

Concentration 

(mg/mL)

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration

(mg/mL) %Recovery 

Me

%Rec

12 1501 6/23/06 Replicate 1 5.21412 3.7571 3.4471 72.1 66

6/23/06 Replicate 2 3.1372 60.2 

12 2400 6/23/06 Replicate 1 95.40212 92.9968 89.8037 97.5 94

6/23/06 Replicate 2 86.6106 90.8 

9 1501 6/26/06 Replicate 1 4.40683 3.5215 3.7021 79.9 84

6/26/06 Replicate 2 3.8828 88.1 

9 24001 6/26/06 Replicate 1 102.98223 99.2757 85.0652 96.4 82

6/26/06 Replicate 2 70.8548 68.8 
MPI Research Study Number 1271-001 

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

Stability Analysis of Dosing Formulations 

RSD - Relative Standard Deviation 

1 These data failed to meet acceptance criterion for recovery (±10%). 4 Initial calculated concentration was based on the average calculated concentration from the Week 3 concentration analyses of do5 These data are the result of a redilution and reanalysis conducted due to the original results failing to meet acceptance criterion fo

Reanalysis confirmed the original results; therefore, the mean of both analyses has been reported. 

Stability 

Interval 

(Day) 

Dose Level 

(mg/kg/day) Sample Identification 

Initial Calculated 

Concentration 

(mg/mL)

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration

(mg/mL) %Recovery 

Me

%Rec

57 1505 7/11/06 Replicate 1 5.81274 1.5039 1.7218 25.9 29

7/11/06 Replicate 2 1.5903 27.4 

7/11/06 Redilution Replicate 1 1.1627 20.0 

7/11/06 Redilution Replicate 2 2.6301 45.2 

57 24001 7/11/06 Replicate 1 101.67584 110.7990 112.7601 109.0 110

7/11/06 Replicate 2 114.7213 112.8 
MPI Research Study Number 1271-001 

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

Stability Analysis of Dosing Formulations 

RSD - Relative Standard Deviation 

1 These data failed to meet acceptance criterion for recovery (±10%). 2 Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from 

homogeneity analysis of dosing formulations. 3 Initial calculated concentration was based on the average calculated concentration from the average of the middle replicates from homogeneity analysis of dosing formulations. 

Stability 

Interval 

(Day) 

Dose Level 

(mg/kg/day) Sample Identification 

Initial Calculated 

Concentration 

(mg/mL)

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration

(mg/mL) %Recovery 

Me

%Rec

21 150 6/23/06 Replicate 1 5.21412 4.9598 4.8212 95.1 92

6/23/06 Replicate 2 4.6827 89.8 

21 24001 6/23/06 Replicate 1 95.40212 76.6087 75.1954 80.3 78

6/23/06 Replicate 2 73.7820 77.3 

18 1501 6/26/06 Replicate 1 4.40683 5.0838 51.1182 115.4 116

6/26/06 Replicate 2 5.1526 116.9 

18 24001 6/26/06 Replicate 1 102.98223 83.5364 83.6652 81.1 81

6/26/06 Replicate 2 83.7941 81.4 
MPI Research Study Number 1271-001 

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

Stability Analysis of Dosing Formulations 

RSD - Relative Standard Deviation 

1 These data failed to meet acceptance criterion for recovery (±10%). 4 Initial calculated concentration was based on the average calculated concentration from the Week 3 concentration analyses of do6 These data are the result of a redilution and reanalysis conducted due to the original results failing to meet acceptance criterion fo

Stability 

Interval 

(Day) 

Dose Level 

(mg/kg/day) Sample Identification 

Initial Calculated 

Concentration 

(mg/mL)

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration

(mg/mL) %Recovery 

Me

%Rec

51 1501 7/11/06 Replicate 1 5.81274 2.5876 2.7896 44.5 48

7/11/06 Replicate 2 2.9916 51.5 

57 1501,6 7/11/06 Redilution Replicate 1 5.81274 1.1075 1.7306 19.1 29

7/11/06 Redilution Replicate 2 2.3537 40.5 

51 24001 7/11/06 Replicate 1 101.67584 116.6508 115.8385 114.7 113

7/11/06 Replicate 2 115.0262 113.1 
MPI Research Study Number 1271-001 

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

Week 1 Concentration Analysis of Dosing Formulations

RSD - Relative Standard Deviation 

NA - Not Applicable/Not Available 

1 These data failed to meet acceptance criterion for recovery (±15%). 

Dose Level 

(mg/kg/day) Sample Identification 

Nominal 

Concentration 

(mg/mL) 

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration 

(mg/mL) 

Average 

Dose Level 

(mg/kg/day) %Recover

0 Replicate 1 0.0 0.0000 0.0000 0.0000 NA 

Replicate 2 0.0000 NA 

1501 Replicate 1 7.50 5.7402 5.4747 109.4940 76.5 

Replicate 2 5.2092 69.5 

6001 Replicate 1 30.00 19.6762 19.8070 396.1400 65.6 

Replicate 2 19.9378 66.5 

24001 Replicate 1 120.00 85.1654 83.0470 1660.9400 71.0 

Replicate 2 80.9286 67.4 
MPI Research Study Number 1271-001 

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

Week 3 Concentration Analysis of Dosing Formulations

RSD - Relative Standard Deviation 

NA - Not Applicable/Not Available 

1 These data failed to meet acceptance criterion for recovery (±15%). 2 These data failed to meet acceptance criterion for recovery (±15%) and RSD (≤10%). 

Dose Level 

(mg/kg/day) Sample Identification 

Nominal 

Concentration 

(mg/mL) 

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration 

(mg/mL) 

Average 

Dose Level 

(mg/kg/day) %Recover

0 Replicate 1 0.0 0.0000 0.0000 0.0000 NA 

Replicate 2 0.0000 NA 

1501 Replicate 1 7.50 5.8237 5.8127 116.2540 77.6 

Replicate 2 5.8018 77.4 

6002 Replicate 1 30.00 15.4342 17.8468 356.9360 51.4 

Replicate 2 20.2594 67.5 

24002 Replicate 1 120.00 109.6681 101.6758 2033.5160 91.4 

Replicate 2 93.6834 78.1 
MPI Research Study Number 1271-001 

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

Week 8 Concentration Analysis of Dosing Formulations

RSD - Relative Standard Deviation 

NA - Not Applicable/Not Available 

1 These data failed to meet acceptance criterion for recovery (±15%). 3 These data are the result of a reinjection conducted due to a failed check standard. 4 These data are the result of a redilution and reanalysis conducted due to the original results failing to meet acceptance criteria for 

RSD (≤10%). Reanalysis confirmed the original results; therefore, the mean of both analyses has been reported. 

Dose Level 

(mg/kg/day) Sample Identification 

Nominal 

Concentration 

(mg/mL) 

Calculated 

Concentration 

(mg/mL) 

Average 

Calculated 

Concentration 

(mg/mL) 

Average 

Dose Level 

(mg/kg/day) %Recover

03 Reinjection Replicate 1 0.0 0.0000 0.0000 0.0000 NA 

Reinjection Replicate 2 0.0000 NA 

1503,4 Reinjection Replicate 1 7.50 2.4108 1.8801 37.6020 32.1 

Reinjection Replicate 2 2.0710 27.6 

Redilution Replicate 1 1.1399 15.2 

Redilution Replicate 2 1.8989 25.3 

6001,3 Reinjection Replicate 1 30.00 23.2112 23.1492 462.9840 77.4 

Reinjection Replicate 2 23.0871 77.0 

24003 Reinjection Replicate 1 120.00 117.5559 119.4088 2388.1760 98.0 

Reinjection Replicate 2 121.2618 101.1 
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 Screening Study of Anthraquinone (CAS # 84-65-1) in Rats 

Record of P Animal Fate and Disposition - MALE 

Group, 

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 

324r 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 

333r terminal necropsy 45 

334 terminal necropsy 48 

335 terminal necropsy 48 

336 terminal necropsy 48 

2400 mg/kg/day

337r 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 Screening Study of Anthraquinone (CAS # 84-65-1) in Rats 

Record of P Animal Fate and Disposition - MALE 

Group, 

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 Screening Study of Anthraquinone (CAS # 84-65-1) in Rats 

Record of P Animal Fate and Disposition - FEMALE 

Group, 

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 

369r terminal necropsy 44 

370 terminal necropsy 48 

371r terminal necropsy 50 

372 terminal necropsy 55 

600 mg/kg/day

373r terminal necropsy 44 

374 found dead 46 

375 found dead 47 

376 found dead 45 

377 terminal necropsy 51 

378r found dead 41 

379 found dead 44 

380 found dead 48 

381r 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 Screening Study of Anthraquinone (CAS # 84-65-1) in Rats 

Record of P Animal Fate and Disposition - FEMALE 

Group, 

Animal Number Fate Day 

2400 mg/kg/day

389 terminal necropsy 49 

390 terminal necropsy 50 

391r found dead 41 

392r terminal necropsy 46 

393 euthanized in extremis 39 

394 found dead 44 

395r 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 358
MPI Research Study Number 1271-001 

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

Individual P Clinical Findings - MALE 

Group, 

Animal Number Day Sign Present 0 mg/kg/day

301 No Abnormalities Detected 1, 3-47 

 Hair discolored, Yellow, Anogenital region 2 

302 No Abnormalities Detected 1-21, 24-47 

 Urine discolored, Orange 22-23 

303 No Abnormalities Detected 1-22 

 Hair sparse, Forefoot/left 23-47  Hair sparse, Forefoot/right 23-47 

304 No Abnormalities Detected 1-47 305 No Abnormalities Detected 1-47 306 No Abnormalities Detected 1-47 307 No Abnormalities Detected 1-47 308 No Abnormalities Detected 1-47 309 No Abnormalities Detected 1-47 310 No Abnormalities Detected 1-47 311 No Abnormalities Detected 1-47 312 No Abnormalities Detected 1-47 

150 mg/kg/day

313 No Abnormalities Detected 1-27, 34, 36-37, 40-47 

 Material around nose, Red 38-39  Urine discolored, Orange 30  Urine discolored, Red 28-29, 31-33, 35 
MPI Research Study Number 1271-001 

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

Individual P Clinical Findings - MALE 

Group, 

Animal Number Day Sign Present 150 mg/kg/day

314 No Abnormalities Detected 1-22 

 Hair sparse, Forelimb/left 23-47  Hair sparse, Forelimb/right 23-47  Urine discolored, Orange 30  Urine discolored, Red 28-29, 31-33, 35 

315 No Abnormalities Detected 1-23, 27-28, 34, 36-47 

 Urine discolored, Orange 24-26, 30  Urine discolored, Red 29, 31-33, 35 

316 No Abnormalities Detected 1-2, 14-23, 27, 34, 36 

 Hair sparse, Forefoot/left 3-13  Hair sparse, Forefoot/right 3-13  Hair sparse, Forelimb/left 37-47  Hair sparse, Forelimb/right 37-47  Urine discolored, Orange 24-26, 30  Urine discolored, Red 28-29, 31-33, 35 

317 No Abnormalities Detected 1-24, 27-28 

 Urine discolored, Orange 25-26, 30  Urine discolored, Red 29, 31-33 

318 No Abnormalities Detected 1-22, 27-28, 34, 36-47 

 Urine discolored, Orange 23-26, 30  Urine discolored, Red 29, 31-33, 35 

319 No Abnormalities Detected 1-23, 27-28, 34, 36-41, 43-47 

 Urine discolored, Orange 24-26, 30  Urine discolored, Red 29, 31-33, 35, 42 

320 No Abnormalities Detected 1-22, 27, 34, 36-41, 45-47 
MPI Research Study Number 1271-001 

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

Individual P Clinical Findings - MALE 

Group, 

Animal Number Day Sign Present 150 mg/kg/day

 Urine discolored, Orange 23-26, 30  Urine discolored, Red 28-29, 31-33, 35, 42-44 

321 No Abnormalities Detected 1-24, 27-30, 34-47 

 Urine discolored, Orange 25-26  Urine discolored, Red 31-33 

322 No Abnormalities Detected 1-24, 27-28 

 Hair discolored, Brown, Forelimb/left 33-47  Hair discolored, Brown, Forelimb/right 33-47  Urine discolored, Orange 25-26, 30  Urine discolored, Red 29, 31-33 

323 No Abnormalities Detected 1-2, 15-24, 27-28, 34, 36-39, 41-47 

 Activity decreased 3-5  Breathing difficult 3-5  Carriage low 4-5  Piloerection 3-14  Posture hunched 3-11  Urine discolored, Orange 25-26, 30  Urine discolored, Red 29, 31-33, 35, 40 

324 r No Abnormalities Detected 1-18, 23-24, 30, 32 

 Hair sparse, Face 33-43  Hair sparse, Ventral surface 35-43  Urine discolored, Orange 19-22, 26  Urine discolored, Red 25, 27-29, 31, 38 

600 mg/kg/day

325 No Abnormalities Detected 1-10, 22-28, 34, 36-47 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Clinical Findings - MALE 

Group, 

Animal Number Day Sign Present 600 mg/kg/day

 Urine discolored, Orange 30  Urine discolored, Red 11-21, 29, 31-33, 35 

326 No Abnormalities Detected 2-10, 17 

 Abrasion(s), Cervical region 27-47  Hair sparse, Cervical region 37-47  Salivation 1  Scabbed area, Thoracic region 20-26  Urine discolored, Orange 23-26, 30  Urine discolored, Red 11-16, 18-21, 29, 31-33, 35-42 

327 No Abnormalities Detected 1-7, 27, 34, 36-39 

 Malocclusion 40-47  Material around eyes, Black, Eye/left 44-47  Material around eyes, Black, Eye/right 44-47  Material around eyes, Red, Eye/left 40-43  Material around eyes, Red, Eye/right 40-43  Urine discolored, Orange 22-26, 28, 30  Urine discolored, Red 8-21, 29, 31-33, 35, 40 

328 No Abnormalities Detected 1-10, 17-18, 27, 34, 36-37 

 Hair sparse, Forelimb/right 38-47  Urine discolored, Orange 22-26, 28, 30  Urine discolored, Red 11-16, 19-21, 29, 31-33, 35 

329 No Abnormalities Detected 1-10, 17-18, 22-24, 27, 34, 36-47 

 Urine discolored, Orange 25-26, 28, 30  Urine discolored, Red 11-16, 19-21, 29, 31-33, 35 

330 No Abnormalities Detected 1-8, 22-24, 27, 34, 36-46 

 Urine discolored, Orange 25-26, 28, 30 
MPI Research Study Number 1271-001 

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

Individual P Clinical Findings - MALE 

Group, 

Animal Number Day Sign Present 600 mg/kg/day

 Urine discolored, Red 9-21, 29, 31-33, 35, 47 

331 No Abnormalities Detected 1-8, 22-24, 27, 34, 36 

 Hair sparse, Forelimb/right 37-47  Urine discolored, Orange 25-26, 28, 30  Urine discolored, Red 9-21, 29, 31-33, 35, 47 

332 No Abnormalities Detected 1-8, 22-24, 34, 36-46 

 Urine discolored, Orange 25-28, 30  Urine discolored, Red 9-21, 29, 31-33, 35, 47 

333 r No Abnormalities Detected 1-5, 19-21, 24, 31, 33-43 

 Urine discolored, Orange 22-23, 25, 27  Urine discolored, Red 6-18, 26, 28-30, 32, 44 

334 No Abnormalities Detected 1-5, 7-8, 22-24, 27-29, 34, 36-47 

 Urine discolored, Brown 6  Urine discolored, Orange 25-26, 30  Urine discolored, Red 9-21, 31-33, 35 

335 No Abnormalities Detected 1-2, 7-10, 22-24, 27-28, 34, 36-47 

 Urine discolored, Brown 3-6  Urine discolored, Orange 25-26, 30  Urine discolored, Red 11-21, 29, 31-33, 35 

336 No Abnormalities Detected 1-8, 22-24, 27, 34, 36-47 

 Urine discolored, Orange 25-26, 28, 30  Urine discolored, Red 9-21, 29, 31-33, 35 

2400 mg/kg/day

337 r No Abnormalities Detected 1-4, 13-14, 18-21, 23, 30, 32-39, 41-43 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Clinical Findings - MALE 

Group, 

Animal Number Day Sign Present 2400 mg/kg/day

 Urine discolored, Orange 22, 24, 26, 40  Urine discolored, Red 5-12, 15-17, 25, 27-29, 31 

338 No Abnormalities Detected 1-8, 17-18, 22-25, 27, 34, 36-47 

 Urine discolored, Orange 26, 28, 30  Urine discolored, Red 9-16, 19-21, 29, 31-33, 35 

339 No Abnormalities Detected 4-7, 22-24, 27, 34, 36-43, 45-47 

 Abrasion(s), Lip 1  Hair discolored, Brown, Anogenital region 2-3  Urine discolored, Orange 25-26, 28, 30, 44  Urine discolored, Red 8-21, 29, 31-33, 35 

340 No Abnormalities Detected 1-10, 22-24, 27 

 Hair discolored, Brown, Ventral surface 30-47  Hair discolored, Yellow, Ventral surface 29  Urine discolored, Orange 25-26, 28, 30, 44  Urine discolored, Red 11-21, 29, 31-33, 35 

341 No Abnormalities Detected 1-11, 22-24, 27-28, 34, 36 

 Hair sparse, Forefoot/left 37-47  Hair sparse, Forefoot/right 37-47  Urine discolored, Orange 25-26, 30, 44  Urine discolored, Red 12-21, 29, 31-33, 35 

342 No Abnormalities Detected 1-8, 22-24, 27, 34, 36-38, 41-43, 45-47 

 Urine discolored, Orange 25-26, 28, 30, 44  Urine discolored, Red 9-21, 29, 31-33, 35, 39-40 

343 No Abnormalities Detected 1-8, 22-24, 27, 34, 36-43, 45-47 

 Urine discolored, Orange 25-26, 28, 30, 44  Urine discolored, Red 9-21, 29, 31-33, 35 
MPI Research Study Number 1271-001 

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

Individual P Clinical Findings - MALE 

Group, 

Animal Number Day Sign Present 2400 mg/kg/day

344 No Abnormalities Detected 1-8, 22-24, 27, 34, 36-43, 45-47 

 Urine discolored, Orange 25-26, 28, 30, 44  Urine discolored, Red 9-21, 29, 31-33, 35 

345 No Abnormalities Detected 1-8, 17-18, 22-24, 27-28, 34, 36-43, 45-47 

 Urine discolored, Orange 25-26, 30, 44  Urine discolored, Red 9-16, 19-21, 29, 31-33, 35 

346 No Abnormalities Detected 1-8, 17, 22-24, 27-28, 34, 36-43, 45-47 

 Urine discolored, Orange 25-26, 30, 44  Urine discolored, Red 9-16, 18-21, 29, 31-33, 35 

347 No Abnormalities Detected 2-8, 22-24, 27, 34, 36-43, 45-46 

 Rales 1  Urine discolored, Orange 25-26, 28, 30, 44  Urine discolored, Red 9-21, 29, 31-33, 35, 47 

348 No Abnormalities Detected 2-4, 6-8, 17, 22-24, 27, 34, 36 

 Material around nose, Black 5  Swelling, Anogenital region 1, 37-47  Urine discolored, Orange 25-26, 28, 30, 44  Urine discolored, Red 9-16, 18-21, 29, 31-33, 35 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Clinical Findings - FEMALE 

Group, 

Animal Number Day Sign Present 0 mg/kg/day

349 No Abnormalities Detected 1-24 350 No Abnormalities Detected 1-21 351 No Abnormalities Detected 1-21 352 No Abnormalities Detected 1-33 353 No Abnormalities Detected 1-5, 7-22 

 Breathing audible 6 

354 No Abnormalities Detected 1-23 355 No Abnormalities Detected 1-23 356 No Abnormalities Detected 1-23 357 No Abnormalities Detected 1-21 358 No Abnormalities Detected 1-22 359 No Abnormalities Detected 1-20 360 No Abnormalities Detected 1-6 

 Lacrimation, Eye/right 18  Malocclusion 7-22  Material around eyes, Red, Eye/right 7-22  Material around nose, Red 7-8  Urine discolored, Orange 22 

150 mg/kg/day

361 No Abnormalities Detected 1-22 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Clinical Findings - FEMALE 

Group, 

Animal Number Day Sign Present 150 mg/kg/day

362 No Abnormalities Detected 1-4, 7-19 

 Breathing audible 5-6  Feces few/absent 20  Posture hunched 20 

363 No Abnormalities Detected 1-20 

 Hair discolored, Yellow, Ventral surface 21-28  Urine discolored, Orange 26 

364 No Abnormalities Detected 1-20 365 No Abnormalities Detected 1-23 366 No Abnormalities Detected 1-4 

 Hair sparse, Abdominal region 5-23 

367 No Abnormalities Detected 1-3, 7-23 

 Urine discolored, Brown 4-6 

368 No Abnormalities Detected 1-21 369 r No Abnormalities Detected 1-4 

 Feces few/absent 5-8  Hair sparse, Ventral surface 5-16  Posture hunched 5-8 

370 No Abnormalities Detected 1-21 371 r No Abnormalities Detected 1-16 372 No Abnormalities Detected 1-28 

 Urine discolored, Orange 30  Urine discolored, Red 29, 31-33 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Clinical Findings - FEMALE 

Group, 

Animal Number Day Sign Present 600 mg/kg/day

373 r No Abnormalities Detected 1-3 

 Feces few/absent 4  Piloerection 4-5  Posture hunched 4-6  Urine discolored, Red 7-15 

374 No Abnormalities Detected 1-2, 7-10, 22 

 Activity decreased 3-6  Piloerection 3-6  Urine discolored, Brown 6  Urine discolored, Red 11-21 

375 No Abnormalities Detected 1-3, 5-10 

 Breathing audible 16  Rales 4  Urine discolored, Red 11-21 

376 No Abnormalities Detected 1-5, 7-10 

 Activity decreased 6  Hair discolored, Yellow, Anogenital region 6  Piloerection 6  Urine discolored, Red 11-21 

377 No Abnormalities Detected 1 

 Activity decreased 3-5  Breathing rapid 4-5  Hair discolored, Brown, Anogenital region 4  Hair sparse, Abdominal region 2-23  Hypersensitive to touch 5-7  Posture hunched 3-5 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Clinical Findings - FEMALE 

Group, 

Animal Number Day Sign Present 600 mg/kg/day

 Urine discolored, Red 15-16, 18-21 

378 r No Abnormalities Detected 1-7, 12, 17 

 Hair discolored, Brown, Anogenital region 18  Urine discolored, Red 8-11, 13-16 

379 No Abnormalities Detected 1-10, 17, 22-25, 27 

 Urine discolored, Orange 26, 28, 30  Urine discolored, Red 11-16, 18-21, 29, 31-33 

380 No Abnormalities Detected 1-11, 17-18, 22-24, 27-28 

 Urine discolored, Orange 25-26, 30  Urine discolored, Red 12-16, 19-21, 29, 31-33 

381 r No Abnormalities Detected 1-2, 5-16, 19-21 

 Rales 3-4  Urine discolored, Orange 22  Urine discolored, Red 17-18 

382 No Abnormalities Detected 1-7, 11-13, 17-18 

 Feces few/absent 9  Posture hunched 8-10  Urine discolored, Red 14-16, 19-21 

383 No Abnormalities Detected 1, 3, 7-10, 17, 22-23 

 Breathing audible 4-6  Hair discolored, Yellow, Anogenital region 2  Material around mouth, Red 4-5  Urine discolored, Red 11-16, 18-21 

384 No Abnormalities Detected 1-7 

 Scabbed area, Dorsal surface 19-21 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Clinical Findings - FEMALE 

Group, 

Animal Number Day Sign Present 600 mg/kg/day

 Swelling, Dorsal surface 8-18  Urine discolored, Red 15-16, 19-21 

2400 mg/kg/day

385 No Abnormalities Detected 1-2, 7-10, 17, 22-24 

 Activity decreased 3-6  Feces discolored, Yellow 27-33  Feces watery 27-33  Material around mouth, Red 27  Material around nose, Red 27  Piloerection 30-33  Posture hunched 27-33  Skin discolored, Pale, Entire body 33  Thin 33  Unkempt appearance 32-33  Urine discolored, Orange 25-26, 30  Urine discolored, Red 11-16, 18-21, 29, 31-33 

386 No Abnormalities Detected 1-11 

 Urine discolored, Red 12-21 

387 No Abnormalities Detected 1-8 

 Urine discolored, Red 9-21 

388 No Abnormalities Detected 1-8, 22 

 Urine discolored, Red 9-21 

389 No Abnormalities Detected 1 

 Activity decreased 3  Hair sparse, Abdominal region 6-18 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Clinical Findings - FEMALE 

Group, 

Animal Number Day Sign Present 2400 mg/kg/day

 Hair sparse, Hind limb/left 6-21  Hair sparse, Inguinal region/right 2-18  Hair sparse, Lumbar region 15-21  Hair sparse, Ventral surface 19-21  Urine discolored, Red 9-21 

390 No Abnormalities Detected 1-4, 6-8, 17, 22-23 

 Activity decreased 5  Urine discolored, Red 9-16, 18-21 

391 r No Abnormalities Detected 1-6, 13 

 Urine discolored, Red 7-12, 14-17 

392 r No Abnormalities Detected 1-3 

 Hair sparse, Hind limb/right 4-17  Urine discolored, Red 5-12, 17 

393 No Abnormalities Detected 1-8, 17 

 Urine discolored, Red 9-16, 18-21 

394 No Abnormalities Detected 1-8, 17-18, 22-24, 27 

 Urine discolored, Orange 25-26, 28, 30  Urine discolored, Red 9-16, 19-21, 29, 31-33 

395 r No Abnormalities Detected 1-4, 13 

 Urine discolored, Red 5-12, 14-16 

396 No Abnormalities Detected 1-8, 17, 22-23 

 Urine discolored, Red 9-16, 18-21 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 0 mg/kg/day

349 No Abnormalities Detected 0-21 350 No Abnormalities Detected 0-21 351 No Abnormalities Detected 0-21 353 No Abnormalities Detected 0-21 354 No Abnormalities Detected 0-21 355 No Abnormalities Detected 0-22 356 No Abnormalities Detected 0-21 357 No Abnormalities Detected 0-21 358 No Abnormalities Detected 0-21 359 No Abnormalities Detected 0-21 360 Malocclusion 0-21 

 Material around eyes, Red, Eye/right 0, 5-14  Urine discolored, Orange 0 

150 mg/kg/day

361 No Abnormalities Detected 0-8, 11-13 

 Hair sparse, Anogenital region 14-22  Rales 16  Urine discolored, Red 9-10 

362 No Abnormalities Detected 7-10, 13, 17-21 

 Hair discolored, Brown, Anogenital region 1-6  Posture hunched 0-2 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 150 mg/kg/day

 Urine discolored, Orange 1-2  Urine discolored, Red 11-12, 14-16 

363 No Abnormalities Detected 20-21 

 Hair discolored, Red, Ventral surface 10-19  Hair discolored, Yellow, Ventral surface 0-9  Urine discolored, Red 3-4, 6-8 

364 No Abnormalities Detected 0-10, 13, 15-22 

 Urine discolored, Red 11-12, 14 

365 No Abnormalities Detected 0-7, 10, 12-22 

 Urine discolored, Red 8-9, 11 

366 Hair sparse, Abdominal region 0-22 

 Urine discolored, Orange 6  Urine discolored, Red 7-9, 11, 13 

367 No Abnormalities Detected 0-5, 10, 12-22 

 Urine discolored, Orange 6  Urine discolored, Red 7-9, 11 

368 No Abnormalities Detected 0-7, 12, 14-21 

 Urine discolored, Orange 8  Urine discolored, Red 9-11, 13 

369 r Hair sparse, Ventral surface 0-22 

 Urine discolored, Orange 9  Urine discolored, Red 10-12, 14-16 

370 No Abnormalities Detected 0-7, 12, 14, 16 

 Hair sparse, Forelimb/left 17-21  Hair sparse, Forelimb/right 17-21 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 150 mg/kg/day

 Urine discolored, Orange 8  Urine discolored, Red 9-11, 13, 15 

371 r No Abnormalities Detected 0-5, 12, 14-17 

 Hair sparse, Forelimb/left 18-24  Hair sparse, Forelimb/right 18-24  Urine discolored, Orange 8  Urine discolored, Red 6-7, 9-11, 13 

600 mg/kg/day

373 r No Abnormalities Detected 1-3, 6-7, 13-15 

 Activity decreased 23  Carriage high 16-17, 22-23  Piloerection 16-23  Skin cold to touch 23  Thin 17-23  Urine discolored, Orange 4-5, 9  Urine discolored, Red 0, 8, 10-12, 16-19 

374 No Abnormalities Detected 0-1, 4-5, 11, 13-21 

 Activity decreased 22  Breathing slow 22  Piloerection 22  Urine discolored, Orange 2-3, 7  Urine discolored, Red 6, 8-10, 12 

375 No Abnormalities Detected 0-2, 12, 14, 19-22 

 Piloerection 23  Skin cold to touch 23  Skin discolored, Pale, Entire body 23 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 600 mg/kg/day

 Urine discolored, Orange 3-6, 8  Urine discolored, Red 7, 9-11, 13, 15-18 

376 No Abnormalities Detected 0-2, 5-6, 12, 14-16, 19-21 

 Feces discolored, Yellow 22  Feces watery 22  Skin cold to touch 22  Urine discolored, Orange 3-4, 8  Urine discolored, Red 7, 9-11, 13, 17-18 

377 Hair sparse, Abdominal region 0-22 

 Urine discolored, Orange 1-2, 4, 6  Urine discolored, Red 5, 7-9, 11-13 

378 r Hair discolored, Brown, Anogenital region 0-14, 19 

 Hair discolored, Red, Anogenital region 15-18, 21  Hair discolored, Yellow, Anogenital region 20  Piloerection 21  Urine discolored, Orange 2, 4, 6  Urine discolored, Red 5, 7-13 

381 r No Abnormalities Detected 1, 8, 10 

 Hair discolored, Brown, Anogenital region 11-12  Hair discolored, Red, Anogenital region 13-22  Urine discolored, Orange 0, 2, 4  Urine discolored, Red 3, 5-7, 9 

382 No Abnormalities Detected 0-2, 5, 12, 14-19 

 Activity decreased 22  Eyelid partially/completely closed, Eye/left 22  Eyelid partially/completely closed, Eye/right 22 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 600 mg/kg/day

 Material around mouth, Red 20-22  Material around nose, Red 20-22  Piloerection 22  Skin cold to touch 22  Urine discolored, Orange 3-4, 6, 8  Urine discolored, Red 7, 9-11, 13 

383 No Abnormalities Detected 0, 3, 10, 12-21 

 Urine discolored, Orange 1-2, 4, 6  Urine discolored, Red 5, 7-9, 11 

384 No Abnormalities Detected 12, 14 

 Hair sparse, Abdominal region 17-22  Hair sparse, Anogenital region 15-22  Hair sparse, Cervical region 15-22  Hair sparse, Forelimb/left 17-22  Hair sparse, Forelimb/right 17-22  Scabbed area, Dorsal surface 0-9  Urine discolored, Orange 3-4, 6, 8  Urine discolored, Red 7, 9-11, 13, 15 

2400 mg/kg/day

386 No Abnormalities Detected 0-2, 5, 12, 14 

 Hair sparse, Forelimb/left 17-21  Hair sparse, Forelimb/right 15-21  Urine discolored, Orange 3-4, 6, 8  Urine discolored, Red 7, 9-11, 13, 15, 18 

387 No Abnormalities Detected 0-2, 5, 12, 14, 16, 19-22 

 Urine discolored, Orange 3-4, 6, 8 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 2400 mg/kg/day

 Urine discolored, Red 7, 9-11, 13, 15, 17-18 

388 No Abnormalities Detected 0-1, 4-5, 11, 13 

 Activity decreased 21  Breathing slow 21  Discharge, Clear, Anus 14-17  Feces watery 18-21  Hair discolored, Brown, Anogenital region 20-21  Piloerection 19-21  Posture hunched 19-21  Skin discolored, Pale, Entire body 20-21  Thin 19-21  Urine discolored, Orange 2-3, 7  Urine discolored, Red 6, 8-10, 12, 14 

389 Hair sparse, Abdominal region 15-20 

 Hair sparse, Hind limb/left 0-22  Hair sparse, Lumbar region 0-10  Hair sparse, Ventral surface 0-10  Material around mouth, Red 22  Material around nose, Red 22  Urine discolored, Orange 6, 8  Urine discolored, Red 7, 9-11 

390 No Abnormalities Detected 0, 3, 10, 12, 14, 17-21 

 Urine discolored, Orange 1-2, 4, 6  Urine discolored, Red 5, 7-9, 11, 13, 15-16 

391 r No Abnormalities Detected 0-2, 5, 12, 14, 16, 19-21 

 Activity decreased 22  Feces watery 22 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 2400 mg/kg/day

 Piloerection 22  Skin cold to touch 22  Skin discolored, Blue, Abdominal region 22  Urine discolored, Orange 3-4, 6, 8  Urine discolored, Red 7, 9-11, 13, 15, 17-18 

392 r Hair sparse, Hind limb/right 0-23 

 Urine discolored, Orange 3-4, 6, 8  Urine discolored, Red 7, 9-11, 13, 15, 20 

393 No Abnormalities Detected 0-2, 12, 14 

 Activity decreased 17  Discharge, Red, Vulva 17  Hair discolored, Red, Abdominal region 17  Hair discolored, Red, Anogenital region 17  Material around mouth, Red 17  Material around nose, Black 15-17  Material in pan/bedding, Red 17  Piloerection 17  Skin cold to touch 17  Skin discolored, Pale, Entire body 17  Urine discolored, Orange 3-6, 8  Urine discolored, Red 7, 9-11, 13 

395 r No Abnormalities Detected 1-6, 13, 15, 17, 20-22 

 Discharge, Red, Vulva 23  Eye discolored, Pale, Eye/left 23  Eye discolored, Pale, Eye/right 23  Material around mouth, Red 23  Material around nose, Red 23  Material in pan/bedding, Red 23 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Clinical Findings 

Group, 

Animal Number Gestation Day Sign Present 2400 mg/kg/day

 Skin cold to touch 23  Skin discolored, Pale, Entire body 23  Urine discolored, Orange 7, 9  Urine discolored, Red 0, 8, 10-12, 14, 16, 18-19 

396 No Abnormalities Detected 0-3, 10, 12, 14 

 Feces soft 21-31  Material around mouth, Red 19-25, 31  Material around nose, Red 17-25, 31  Thin 23-31  Urine discolored, Orange 4, 6  Urine discolored, Red 5, 7-9, 11, 13, 15-16, 18 
MPI Research Study Number 1271-001 

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

Individual P Lactation Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

349 No Abnormalities Detected 0-4 350 No Abnormalities Detected 0-4 351 No Abnormalities Detected 0-4 353 No Abnormalities Detected 0-4 354 No Abnormalities Detected 0-4 355 No Abnormalities Detected 0-4 356 No Abnormalities Detected 0-4 357 No Abnormalities Detected 0-4 358 No Abnormalities Detected 0-4 359 No Abnormalities Detected 0-4 360 Malocclusion 0-4 

 Material around eyes, Red, Eye/right 1-3 

150 mg/kg/day

361 Hair sparse, Anogenital region 0-4 362 No Abnormalities Detected 0-4 363 No Abnormalities Detected 0-4 364 No Abnormalities Detected 0-4 365 No Abnormalities Detected 0-4 
MPI Research Study Number 1271-001 

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

Individual P Lactation Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

366 Hair sparse, Abdominal region 0-4 367 No Abnormalities Detected 0-4 368 No Abnormalities Detected 0-4 369 r No Abnormalities Detected 4 

 Hair sparse, Ventral surface 0-3 

370 Hair sparse, Forelimb/left 0-4 

 Hair sparse, Forelimb/right 0-4 

600 mg/kg/day

373 r Activity decreased 0-3 

 Carriage high 0-3  Piloerection 0-4  Skin cold to touch 0-3  Thin 0-4 

375 Activity decreased 0 

 Discharge, Red, Vulva 0  Eye discolored, Pale, Eye/left 0  Eye discolored, Pale, Eye/right 0  Hair discolored, Red, Anogenital region 0  Piloerection 0  Skin cold to touch 0  Skin discolored, Pale, Entire body 0 

376 Discharge, Red, Vulva 0 

 Feces watery 0  Hair discolored, Red, Anogenital region 0 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 600 mg/kg/day

 Skin cold to touch 0  Skin discolored, Pale, Entire body 0 

377 No Abnormalities Detected 4 

 Carriage high 0-3  Hair sparse, Abdominal region 0-3  Material around nose, Red 0-1  Thin 0-3 

380 Carriage high 0-2 

 Skin cold to touch 0-2  Thin 0-2  Tremors 1-2 

381 r No Abnormalities Detected 0-2 382 Activity decreased 0-1 

 Breathing shallow 1  Hair discolored, Red, Abdominal region 0-1  Hair discolored, Red, Anogenital region 0-1  Material around mouth, Red 0-1  Material around nose, Red 0-1  Piloerection 0-1  Skin cold to touch 0-1 

383 No Abnormalities Detected 0-4 384 Hair sparse, Abdominal region 0-3 

 Hair sparse, Anogenital region 0-3  Hair sparse, Cervical region 0-3  Hair sparse, Forelimb/left 0-4  Hair sparse, Forelimb/right 0-4 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 2400 mg/kg/day

387 No Abnormalities Detected 0-4 

389 No Abnormalities Detected 1-4 

 Activity decreased 0  Breathing slow 0  Hair sparse, Hind limb/left 0  Material around mouth, Red 0  Material around nose, Red 0  Piloerection 0  Skin cold to touch 0  Skin discolored, Pale, Entire body 0 

390 No Abnormalities Detected 0, 4 

 Thin 1-3 

392 r Activity decreased 0-2 

 Eye discolored, Pale, Eye/left 0-2  Eye discolored, Pale, Eye/right 0-2  Feces watery 0-3  Hair sparse, Hind limb/right 0-4  Piloerection 0-3  Skin discolored, Pale, Entire body 0-2  Thin 0-3 

rReplacement animal 
1271-001 

Appendix E 

Individual P Body Weight Values 

Page 159 of 358
MPI Research Study Number 1271-001 

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

Individual P Body Weight Values, g - MALE 

Group, 

Animal Week of Study Number 1 2 3 4 5 6 70 mg/kg/day

301 273 324 372 407 451 501 529 302 266 325 385 422 463 507 532 303 263 295 331 356 383 409 433 304 267 303 351 375 403 451 471 305 260 290 324 347 352 383 415 306 266 308 351 389 423 458 481 307 247 299 332 359 388 415 435 308 252 299 346 371 413 441 472 309 271 325 367 407 424 462 497 310 261 317 372 403 444 475 507 311 244 281 320 344 368 400 411 312 253 296 336 358 393 430 449 

150 mg/kg/day

313 265 289 326 348 380 402 418 314 262 305 341 365 399 431 444 315 266 311 363 401 425 467 482 316 267 306 350 384 414 445 471 317 264 302 342 365 390 NDC 318 254 290 337 355 389 430 437 319 262 296 336 350 375 399 419 320 257 289 323 354 376 402 409 321 257 306 346 382 401 431 447 322 257 300 333 364 372 426 443 323 264 241 275 328 372 411 430 

324r 306 351 373 407 429 444 NDC 

NDC - No data collected refer to record of fate 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Body Weight Values, g - MALE 

Group, 

Animal Week of Study Number 1 2 3 4 5 6 7600 mg/kg/day

325 258 289 322 342 360 395 410 326 267 292 330 358 386 423 436 327 255 286 328 349 383 408 370 328 255 282 317 344 366 392 406 329 250 285 319 343 383 414 431 330 268 295 322 352 376 410 422 331 269 298 328 345 376 407 405 332 256 287 327 364 384 423 436 

333r 308 344 376 401 432 449 NDC 

334 247 279 313 342 377 408 423 335 270 305 341 365 407 428 453 336 254 280 299 302 334 365 383 

2400 mg/kg/day

337r 300 355 382 416 452 475 NDC 

338 263 295 339 363 393 422 444 339 263 282 314 340 375 404 423 340 256 280 305 325 340 368 391 341 266 295 336 382 402 440 457 342 253 270 297 324 350 380 396 343 254 272 316 326 355 381 401 344 266 300 339 371 397 451 475 345 265 299 327 363 387 416 429 346 256 295 338 379 401 430 461 347 248 267 293 314 331 363 372 348 256 288 332 350 376 407 424 

NDC - No data collected refer to record of fate 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Body Weight Values, g - FEMALE 

Group, 

Animal Week of Study Number 1 2 3 4 50 mg/kg/day

349 207 216 244 254 350 187 200 217 351 199 243 262 352 184 202 214 233 249 353 204 211 236 254 354 193 212 219 234 355 197 217 226 243 356 207 219 230 250 357 185 200 215 358 202 226 238 255 359 190 206 218 360 190 220 235 263 

150 mg/kg/day

361 197 215 217 228 362 196 213 211 363 199 202 216 219 364 185 193 198 365 192 203 216 223 366 203 212 226 234 367 200 225 218 232 368 187 195 207 

369r 188 212 

370 193 207 214 

371r 198 202 

372 188 191 199 214 247 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Body Weight Values, g - FEMALE 

Group, 

Animal Week of Study Number 1 2 3 4 5600 mg/kg/day

373r 178 162 

374 200 202 206 218 375 192 181 190 376 205 196 210 377 195 186 191 185 

378r 193 187 193 

379 202 190 214 212 233 380 196 202 214 187 230 

381r 203 224 236 

382 210 171 193 383 187 204 219 228 384 188 186 210 

2400 mg/kg/day

385 198 203 206 215 186 386 214 214 232 387 213 215 216 388 200 201 215 207 389 188 188 209 390 193 194 200 213 

391r 226 235 

392r 221 223 

393 198 188 199 394 196 210 224 225 217 

395r 203 229 

396 191 187 200 208 

r Replacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Body Weight Values, g 

Group, 

Animal Day of Gestation Number 0 7 14 200 mg/kg/day

349 263 294 314 383 350 228 265 307 388 351 276 327 377 469 353 259 297 335 434 354 244 273 304 388 355 248 281 316 388 356 249 286 317 406 357 231 275 313 380 358 255 299 332 405 359 222 262 296 364 360 263 318 355 450 
MPI Research Study Number 1271-001 

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

Individual P Gestation Body Weight Values, g 

Group, 

Animal Day of Gestation Number 0 7 14 20150 mg/kg/day

361 217 247 276 345 362 186 230 270 319 363 204 245 273 302 364 211 250 271 339 365 225 260 301 374 366 232 263 297 350 367 241 272 315 389 368 212 244 283 340 

369r 226 266 304 363 

370 228 252 292 354 

371r x NP 217 246 241 245 

600 mg/kg/day

373r 164 189 200 200 

374 PIDE 221 234 289 358 375 197 234 265 316 376 PIDE 214 250 279 330 377 198 210 238 277 

378r DP 196 216 247 258 

381r 248 260 288 313 

382 212 240 275 295 383 242 274 291 366 384 205 232 259 320 

DP - Died pregnant NP - Not pregnant 

PIDE - Pregnant, incomplete delivery 

x - Excluded from summary statistics 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Body Weight Values, g 

Group, 

Animal Day of Gestation Number 0 7 14 202400 mg/kg/day

386 DP 232 268 275 342 387 243 256 282 296 388 DP 215 259 254 229 389 210 251 281 320 390 226 240 266 337 

391r DP 242 262 298 352 

392r 242 264 281 328 

393 PEM 222 226 242 NDC 

395r DP 224 241 261 305 

396 ALR 224 241 256 228 

ALR - All resorptions 

DP - Died pregnant 

NDC - No data collected refer to record of fate 

PEM - Pregnant euthanized moribund 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Body Weight Values, g 

Group, 

Animal Day of Lactation Number 0 40 mg/kg/day

349 304 330 350 283 301 351 349 363 353 323 318 354 267 309 355 283 296 356 294 298 357 289 291 358 307 317 359 277 295 360 334 341 

150 mg/kg/day

361 219 180 362 264 252 363 240 251 364 227 273 365 275 306 366 240 273 367 290 309 368 247 258 

369r 259 315 

370 260 273 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Body Weight Values, g 

Group, 

Animal Day of Lactation Number 0 4600 mg/kg/day

373r 153 162 

374 PIDE NDC 375 199 NDC 376 PIDE 211 NDC 377 180 209 380 186 NDC 

381r 228 NDC 

382 209 NDC 383 247 227 384 203 216 

2400 mg/kg/day

387 229 221 389 246 270 390 232 235 

392r 204 223 

NDC - No data collected refer to record of fate 

PIDE - Pregnant, incomplete delivery 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual Postmating Body Weight Values, g - FEMALE 

Group, 

Animal Week of Study Number 6 7 8

 NA- Not Applicable/Not Available 

0 mg/kg/day

352 NP 253 252 265 

150 mg/kg/day

371r NP NA 242 243 

372 NP 242 250 NDC 

600 mg/kg/day

380 PNMD 264 306 

NDC - No data collected refer to record of fate 

NP - Not pregnant 

PNMD - Pregnant no mating date 

rReplacement animal 
1271-001 

Appendix F 

Individual P Body Weight Change Values 

Page 170 of 358
MPI Research Study Number 1271-001 

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

Individual P Gestation Body Weight Change Values, g 

Group, 

Animal Day of Gestation Number 0-7 7-14 14-20 0-200 mg/kg/day

349 31 20 69 120 350 37 42 81 160 351 51 50 92 193 353 38 38 99 175 354 29 31 84 144 355 33 35 72 140 356 37 31 89 157 357 44 38 67 149 358 44 33 73 150 359 40 34 68 142 360 55 37 95 187 
MPI Research Study Number 1271-001 

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

Individual P Gestation Body Weight Change Values, g 

Group, 

Animal Day of Gestation Number 0-7 7-14 14-20 0-20150 mg/kg/day

361 30 29 69 128 362 44 40 49 133 363 41 28 29 98 364 39 21 68 128 365 35 41 73 149 366 31 34 53 118 367 31 43 74 148 368 32 39 57 128 

369r 40 38 59 137 

370 24 40 62 126 

371r x NP 29 -5 4 28 

600 mg/kg/day

373r 25 11 0 36 

374 PIDE 13 55 69 137 375 37 31 51 119 376 PIDE 36 29 51 116 377 12 28 39 79 

378r DP 20 31 11 62 

381r 12 28 25 65 

382 28 35 20 83 383 32 17 75 124 384 27 27 61 115 

DP - Died pregnant NP - Not pregnant 

PIDE - Pregnant, incomplete delivery 

x - Excluded from summary statistics 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Body Weight Change Values, g 

Group, 

Animal Day of Gestation Number 0-7 7-14 14-20 0-202400 mg/kg/day

386 DP 36 7 67 110 387 13 26 14 53 388 DP 44 -5 -25 14 389 41 30 39 110 390 14 26 71 111 

391r DP 20 36 54 110 

392r 22 17 47 86 

393 PEM 4 16 NDC 

395r DP 17 20 44 81 

396 ALR 17 15 -28 4 

ALR - All resorptions 

DP - Died pregnant 

PEM - Pregnant euthanized moribund 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Body Weight Change Values, g 

Group, 

Animal Day of Lactation Number 0-4 0 mg/kg/day

349 26 350 18 351 14 353 -5 354 42 355 13 356 4 357 2 358 10 359 18 360 7 

150 mg/kg/day

361 -39 362 -12 363 11 364 46 365 31 366 33 367 19 368 11 

369r 56 

370 13 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Body Weight Change Values, g 

Group, 

Animal Day of Lactation Number 0-4 600 mg/kg/day

373r 9 

377 29 383 -20 384 13 

2400 mg/kg/day

387 -8 389 24 390 3 

392r 19 

rReplacement animal 
1271-001 

Appendix G 

Individual P Food Consumption Values 

Page 176 of 358
MPI Research Study Number 1271-001 

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

Individual P Food Consumption Values, g/animal/day - MALE 

Group, 

Animal Study Interval (Week) Number 1-2 2-3 5-6 6-70 mg/kg/day

301 29.0 28.6 32.1 31.7302 29.6 31.4 35.1 31.7303 24.0 24.4 24.7 26.0304 27.0 28.4 32.3 29.9305 25.3 24.0 27.0 24.7306 24.1 25.7 27.1 27.1307 26.4 26.7 25.7 26.1308 26.1 28.1 27.7 28.0309 29.1 29.3 29.6 31.6310 27.7 29.7 28.1 29.9311 23.1 24.4 23.1 21.4312 26.9 29.7 29.7 28.9

150 mg/kg/day

313 21.1 24.0 24.1 23.3314 24.9 27.1 27.7 25.9315 23.4 27.3 27.1 26.7316 23.7 26.6 28.7 25.6317 23.3 26.3 NDC318 22.7 26.3 27.0 24.6319 22.0 23.9 25.9 22.9320 23.3 24.6 24.7 24.7321 24.3 27.1 27.4 28.0322 23.0 24.9 27.9 25.7323 11.9 18.3 28.1 25.4

324r 25.4 29.04 27.9 26.0

4 NDC - No data collected refer to record of fate Food consumption calculated using 4 days 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Food Consumption Values, g/animal/day - MALE 

Group, 

Animal Study Interval (Week) Number 1-2 2-3 5-6 6-7600 mg/kg/day

325 23.6 27.0 25.0 25.4326 22.0 23.9 25.4 25.9327 22.4 27.7 24.9 17.1328 21.4 24.9 23.9 24.1329 21.1 25.4 27.6 26.3330 23.1 26.1 27.0 27.0331 24.4 25.0 25.0 25.0332 21.3 27.0 27.7 26.6

333r 25.6 25.5e 26.9 25.6

334 22.9 25.1 27.9 28.7335 22.7 25.6 25.4 25.7336 20.0 20.4 22.7 21.7

2400 mg/kg/day

337r 26.1 29.04 30.7 26.6

338 22.4 28.6 28.4 25.9339 22.3 23.9 27.4 23.3340 22.9 24.7 27.6 24.1341 21.7 28.7 28.0 28.7342 20.1 24.9 28.0 24.0343 21.6 27.1 32.1 30.9344 25.3 28.7 33.4 30.7345 23.1 26.7 27.6 26.0346 23.6 27.4 29.4 29.7347 18.7 21.4 24.0 25.4348 22.1 26.9 28.3 26.7

4Food consumption calculated using 4 days eExcluded from the mean 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Premating Food Consumption Values, g/animal/day - FEMALE 

Group, 

Animal Week of Study Number 1-2 2-30 mg/kg/day

349 18.7 20.1 350 16.0 17.7 351 21.3 23.3 352 16.0 17.9 353 18.0 19.1 354 14.3 15.9 355 19.3 19.1 356 17.7 18.6 357 15.3 18.1 358 19.3 21.9 359 17.9 18.1 360 20.9 22.3 

150 mg/kg/day

361 15.9 15.0 362 16.3 17.0 363 11.9 18.7 364 13.9 15.0 365 14.7 18.3 366 15.1 17.1 367 16.6 15.3 368 13.4 17.1 

369r 11.3 21.8 

370 15.7 18.7 

371r 14.4 15.8 

372 13.3 16.4 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Premating Food Consumption Values, g/animal/day - FEMALE 

Group, 

Animal Week of Study Number 1-2 2-3600 mg/kg/day

373r 9.0 13.2 

374 10.9 15.4 375 11.7 14.3 376 10.0 19.1 377 10.7 16.6 

378r 13.0 16.0 

379 8.9 15.0 380 13.0 18.3 

381r 12.1 

382 7.7 19.9 383 11.0 17.4 384 12.1 18.1 

2400 mg/kg/day

385 11.1 19.4 386 13.0 21.1 387 13.1 17.0 388 10.9 18.0 389 12.0 18.6 390 10.7 17.4 

391r 15.1 19.0 

392r 15.6 18.5 

393 39.6e 16.1 

394 13.6 19.9 

395r 16.6 15.5 

396 10.3 16.7 

eExcluded from the mean 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Food Consumption Values, g/animal/day 

Group, 

Animal Day of Gestation Number 0-7 7-14 14-20 0-200 mg/kg/day

349 21.9 23.7 25.0 23.5 350 22.3 23.4 27.2 24.2 351 28.9 31.1 30.7 30.2 353 25.1 26.3 31.8 27.6 354 21.3 20.7 26.8 22.8 355 24.7 23.6 26.7 24.9 356 25.1 24.7 27.7 25.8 357 24.9 24.6 26.2 25.2 358 27.0 28.0 27.8 27.6 359 22.4 24.1 26.2 24.2 360 29.9 31.6 32.2 31.2 
MPI Research Study Number 1271-001 

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

Individual P Gestation Food Consumption Values, g/animal/day 

Group, 

Animal Day of Gestation Number 0-7 7-14 14-20 0-20150 mg/kg/day

361 18.9 18.9 22.7 20.0 362 19.3 26.1 23.0 22.8 363 19.1 20.9 14.0 18.2 364 19.0 18.9 24.2 20.5 365 22.7 23.9 27.5 24.6 366 21.1 20.0 22.7 21.2 367 22.1 23.7 26.5 24.0 368 20.9 18.1 23.8 20.8 

369r 20.4 22.4 23.3 22.0 

370 20.7 23.1 25.3 23.0 

371r x NP 19.7 16.4 15.5 17.3 

600 mg/kg/day

373r 15.1 14.9 11.8 14.1 

374 PIDE 18.3 22.6 25.2 21.9 

375 20.7 20.7 22.0 21.1 376 PIDE 22.3 22.0 22.5 22.3 377 15.4 14.6 18.0 15.9 

378r DP 18.4 16.7 15.2 16.9 

381r 19.0 22.3 21.7 21.0 

382 18.9 20.9 18.8 19.6 383 23.1 18.9 26.0 22.5 384 20.0 20.7 22.7 21.1 

DP - Died pregnant NP - Not pregnant 

PIDE - Pregnant, incomplete delivery 

x - Excluded from summary statistics 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Gestation Food Consumption Values, g/animal/day 

Group, 

Animal Day of Gestation Number 0-7 7-14 14-20 0-202400 mg/kg/day

386 DP 22.4 19.7 23.2 21.7 387 17.3 18.3 16.8 17.5 388 DP 21.4 18.1 8.2 16.3 389 21.3 20.4 20.0 20.6 390 19.3 21.3 23.5 21.3 

391r DP 20.7 22.3 24.2 22.3 

392r 22.6 19.7 22.0 21.4 

393 PEM 18.7 22.6 NDC 

395r DP 16.4 20.3 23.8 20.0 

396 ALR 19.7 18.4 13.0 17.3 

ALR - All resorptions 

DP - Died pregnant 

NDC - No data collected refer to record of fate 

PEM - Pregnant euthanized moribund 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Food Consumption Values, g/animal/day 

Group, 

Animal Day of Lactation Number 0-4 0 mg/kg/day

349 38.0 350 32.8 351 43.5 353 30.8 354 35.5 355 39.8 356 44.8 357 39.8 358 30.5 359 36.5 360 36.3 

150 mg/kg/day

361 0.5e 

362 39.5 363 23.8 364 28.3 365 41.3 366 30.0 367 35.5 368 24.5 

369r 40.3 

370 28.3 

eExcluded from the mean 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Lactation Food Consumption Values, g/animal/day 

Group, 

Animal Day of Lactation Number 0-4 600 mg/kg/day

373r 9.5 

377 15.0 383 11.3 384 13.3 

2400 mg/kg/day

387 17.5 389 31.8 390 8.3 

392r 14.5 

rReplacement animal 
1271-001 

Appendix H 

Pathology Interpretation 

Page 186 of 358
1271-001 

MPI Research Study Number 1271-001 

An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone 

(CAS # 84-65-1) in Rats 

Pathology Interpretation 

Macroscopic 

Test article-related macroscopic changes were 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-related response, and were considered to be 

incidental. 

Organ Weights 

No test article-related organ weight changes 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 controls, but the relative increases were caused by 

decreased body weight and were not 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 considered 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
187

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 358
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

 Body weight Epididymides Epididymides/ Prostate gl Prostate gl/ Sem. ves. Sem. ves./ Te

Group, BWt BWt BWt 

Animal 

Number g g % g % g % 

0 mg/kg/day301 514 1.146 0.2230 0.409 0.0796 1.682 0.3272 302 553 1.189 0.2150 0.841 0.1521 2.130 0.3852 303 434 1.275 0.2938 1.059 0.2440 1.612 0.3714 304 469 1.102 0.2350 0.674 0.1437 1.643 0.3503 305 418 1.092 0.2612 0.534 0.1278 1.966 0.4703 306 489 1.282 0.2622 0.423 0.0865 1.698 0.3472 307 444 1.172 0.2640 0.632 0.1423 1.648 0.3712 308 472 1.299 0.2752 0.951 0.2015 1.662 0.3521 309 514 1.445 0.2811 1.071 0.2084 1.549 0.3014 310 512 1.217 0.2377 0.853 0.1666 2.122 0.4145 

0.4661 

311 312 

416 454 

1.156 1.277 

0.2779 0.2813 

0.689 0.596 

0.1656 0.1313 

1.939 1.963 0.4324 

eExcluded from the mean 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

Testis, rt Testis, rt/

Group, BWt Animal 

Number g % 

0 mg/kg/day301 1.512 0.2942 302 1.671 0.3022 303 1.542 0.3553 304 1.589 0.3388 305 1.761 0.4213 306 1.996 0.4082 307 1.580 0.3559 308 1.736 0.3678 309 1.654 0.3218 310 1.735 0.3389 311 1.748 0.4202 312 1.670 0.3678 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

 Body weight Epididymides Epididymides/ Prostate gl Prostate gl/ Sem. ves. Sem. ves./ Te

Group, BWt BWt BWt 

Animal 

Number g g % g % g % 

150 mg/kg/day313 NA 1.081 NA 0.436 NA 1.863 NA 314 451 1.152 0.2554 0.620 0.1375 2.057 0.4561 315 484 1.122 0.2318 0.731 0.1510 1.691 0.3494 316 485 1.518 0.3130 0.855 0.1763 1.676 0.3456 318 449 1.168 0.2601 0.610 0.1359 1.884 0.4196 319 429 1.047 0.2441 0.544 0.1268 1.533 0.3573 320 418 1.249 0.2988 0.530 0.1268 1.661 0.3974 321 449 1.126 0.2508 0.575 0.1281 1.831 0.4078 322 452 1.246 0.2757 0.674 0.1491 2.182 0.4827 323 434 1.240 0.2857 0.543 0.1251 2.237 0.5154 

r 324 443 1.208 0.2727 0.701 0.1582 1.762 0.3977 

NA - Not Applicable/Not Available 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

Testis, rt Testis, rt/

Group, BWt Animal 

Number g % 

150 mg/kg/day313 1.650 NA 314 1.565 0.3470 315 1.793 0.3705 316 1.951 0.4023 318 1.780 0.3964 319 1.591 0.3709 320 1.829 0.4376 321 1.790 0.3987 322 1.781 0.3940 323 1.832 0.4221 

r 324 1.687 0.3808 

NA - Not Applicable/Not Available 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

 Body weight Epididymides Epididymides/ Prostate gl Prostate gl/ Sem. ves. Sem. ves./ Te

Group, BWt BWt BWt 

Animal 

Number g g % g % g % 

600 mg/kg/day325 412 1.010 0.2451 0.298 0.0723 1.376 0.3340 326 439 1.126 0.2565 0.522 0.1189 2.094 0.4770 327 400 1.069 0.2673 0.352 0.0880 1.886 0.4715 328 407 1.097 0.2695 0.674 0.1656 1.921 0.4720 329 434 1.158 0.2668 0.583 0.1343 1.910 0.4401 330 427 1.225 0.2869 0.620 0.1452 1.695 0.3970 331 413 1.212 0.2935 0.676 0.1637 2.227 0.5392 332 447 1.134 0.2537 0.595 0.1331 2.058 0.4604 

r 333 447 1.150 0.2573 0.455 0.1018 1.629 0.3644 

334 443 1.191 0.2688 0.656 0.1481 2.095 0.4729 335 457 1.169 0.2558 0.865 0.1893 1.918 0.4197 336 388 1.251 0.3224 0.757 0.1951 1.762 0.4541 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

Testis, rt Testis, rt/

Group, BWt Animal 

Number g % 

600 mg/kg/day325 1.549 0.3760 326 1.659 0.3779 327 1.994 0.4985 328 1.771 0.4351 329 1.512 0.3484 330 1.881 0.4405 331 2.009 0.4864 332 1.751 0.3917 

r 333 1.785 0.3993 

334 1.786 0.4032 335 1.647 0.3604 336 1.934 0.4985 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

 Body weight Epididymides Epididymides/ Prostate gl Prostate gl/ Sem. ves. Sem. ves./ Te

Group, BWt BWt BWt 

Animal 

Number g g % g % g % 

2400 mg/kg/day

r 337 469 1.221 0.2603 0.806 0.1719 1.979 0.4220 

338 447 1.143 0.2557 0.573 0.1282 1.916 0.4286 339 429 1.173 0.2734 0.529 0.1233 2.176 0.5072 340 393 1.214 0.3089 0.531 0.1351 2.144 0.5455 341 462 1.275 0.2760 0.553 0.1197 1.699 0.3677 342 390 1.164 0.2985 0.463 0.1187 1.986 0.5092 343 388 1.166 0.3005 0.664 0.1711 2.031 0.5235 344 478 1.035 0.2165 0.583 0.1220 1.937 0.4052 345 439 1.225 0.2790 0.831 0.1893 2.160 0.4920 346 463 1.177 0.2542 0.558 0.1205 1.672 0.3611 347 375 1.201 0.3203 0.496 0.1323 1.958 0.5221 348 429 1.153 0.2688 0.548 0.1277 1.462 0.3408 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - MALE 

Terminal 

Testis, rt Testis, rt/

Group, BWt Animal 

Number g % 

2400 mg/kg/day

r 337 1.769 0.3772 

338 1.698 0.3799 339 1.840 0.4289 340 1.633 0.4155 341 1.855 0.4015 342 1.733 0.4444 343 1.780 0.4588 344 1.630 0.3410 345 1.710 0.3895 346 1.638 0.3538 347 1.969 0.5251 348 1.882 0.4387 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - FEMALE 

Terminal 

 Body weight Ovaries Ovaries/BWt Uterus w/ Uterus w/

Group, cervix cervix/BWt 

Animal 

Number g g % g % 

0 mg/kg/day349 319 0.149 0.0467 0.949 0.2975 350 288 0.153 0.0531 1.087 0.3774 351 348 0.198 0.0569 0.936 0.2690 

e e e e e 352 261 0.098 0.0375 0.977 0.3743

353 304 0.120 0.0395 0.698 0.2296 354 295 0.124 0.0420 0.667 0.2261 355 287 0.139 0.0484 0.682 0.2376 356 289 0.135 0.0467 0.762 0.2637 357 284 0.090 0.0317 0.686 0.2415 358 304 0.118 0.0388 0.760 0.2500 359 360 

280 327 

0.132 0.146 

0.0471 0.0446 

0.730 0.908 

0.2607 0.2777 

eExcluded from the mean 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - FEMALE 

Terminal 

 Body weight Ovaries Ovaries/BWt Uterus w/ Uterus w/

Group, cervix cervix/BWt 

Animal 

Number g g % g % 

150 mg/kg/day361 177 0.102 0.0576 0.481 0.2718 362 242 0.125 0.0517 0.530 0.2190 363 234 0.122 0.0521 0.779 0.3329 364 262 0.125 0.0477 0.677 0.2584 365 296 0.104 0.0351 0.640 0.2162 366 268 0.153 0.0571 0.528 0.1970 367 301 0.125 0.0415 0.739 0.2455 368 253 0.105 0.0415 0.833 0.3292 

r 369 309 0.144 0.0466 0.865 0.2799 

370 265 0.127 0.0479 0.705 0.2660 

r e e e e e 371 240 0.129 0.0538 0.500 0.2083

e e e e e 372 217 0.116 0.0535 0.303 0.1396

eExcluded from the mean 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - FEMALE 

Terminal 

 Body weight Ovaries Ovaries/BWt Uterus w/ Uterus w/

Group, cervix cervix/BWt 

Animal 

Number g g % g % 

600 mg/kg/day

r 373 160 0.090 0.0563 0.374 0.2338 

377 204 0.122 0.0598 0.572 0.2804 383 224 0.119 0.0531 0.733 0.3272 384 210 0.135 0.0643 0.578 0.2752 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Organ Weight Values - FEMALE 

Terminal 

 Body weight Ovaries Ovaries/BWt Uterus w/ Uterus w/

Group, cervix cervix/BWt 

Animal 

Number g g % g % 

2400 mg/kg/day387 215 0.076 0.0353 0.641 0.2981 389 255 0.118 0.0463 0.644 0.2525 390 227 0.117 0.0515 0.791 0.3485 

r 392 214 0.086 0.0402 0.500 0.2336 

396 201 0.059 0.0294 0.347 0.1726 

rReplacement animal 
1271-001 

Appendix K 

Individual P Animal Data Record: Pathology 

Page 205 of 358
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 0 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

301 S Macroscopic 

all tissues - within normal limits 

301 S Microscopic 

epididymides - within normal limits prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 

302 S Macroscopic 

all tissues - within normal limits 

302 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

303 S Macroscopic 

all tissues - within normal limits 

303 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

304 S Macroscopic 

all tissues - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 304 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

305 S Macroscopic 

all tissues - within normal limits 

305 S Microscopic 

epididymides - within normal limits prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 

306 S Macroscopic 

all tissues - within normal limits 

306 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

307 S Macroscopic 

all tissues - within normal limits 

307 S Microscopic 

epididymides - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 307 S Microscopic 

prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 

308 S Macroscopic 

all tissues - within normal limits 

308 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

309 S Macroscopic 

all tissues - within normal limits 

309 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

310 S Macroscopic 

all tissues - within normal limits 

310 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 310 S Microscopic 

seminal vesicles - within normal limits testes - within normal limits 

311 S Macroscopic 

all tissues - within normal limits 

311 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

312 S Macroscopic 

all tissues - within normal limits 

312 S Microscopic 

epididymides - within normal limits prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 150 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 313 S Macroscopic 

all tissues - within normal limits 

314 S Macroscopic 

all tissues - within normal limits 

315 S Macroscopic 

all tissues - within normal limits 

316 S Macroscopic 

all tissues - within normal limits 

317 D Macroscopic 

thyroid gland - discoloration, red, bilateral, mild 

318 S Macroscopic 

all tissues - within normal limits 

319 S Macroscopic 

all tissues - within normal limits 

320 S Macroscopic 

all tissues - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 150 mg/kg/day321 S Macroscopic 

all tissues - within normal limits 

322 S Macroscopic 

all tissues - within normal limits 

323 S Macroscopic 

cavity, thoracic - adhesion, mild 

all thoracic organs adhered to one another 

324r S Macroscopic 

all tissues - within normal limits 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 600 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 325 S Macroscopic 

all tissues - within normal limits 

326 S Macroscopic 

skin - abrasion, right lateral neck, mild 

corresponds to antemortem observation (ab

327 S Macroscopic 

all tissues - within normal limits 

328 S Macroscopic 

all tissues - within normal limits 

329 S Macroscopic 

all tissues - within normal limits 

330 S Macroscopic 

all tissues - within normal limits 

331 S Macroscopic 

all tissues - within normal limits 

332 S Macroscopic 

all tissues - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 600 mg/kg/day333r S Macroscopic 

all tissues - within normal limits 

334 S Macroscopic 

all tissues - within normal limits 

335 S Macroscopic 

all tissues - within normal limits 

336 S Macroscopic 

all tissues - within normal limits 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day337r S Macroscopic 

all tissues - within normal limits 

337 S Microscopic 

epididymides - within normal limits prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 

338 S Macroscopic 

all tissues - within normal limits 

338 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

339 S Macroscopic 

all tissues - within normal limits 

339 S Microscopic 

epididymides - within normal limits prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 

340 S Macroscopic 

all tissues - within normal limits 

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 340 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

341 S Macroscopic 

all tissues - within normal limits 

341 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

342 S Macroscopic 

all tissues - within normal limits 

342 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

343 S Macroscopic 

all tissues - within normal limits 

343 S Microscopic 

epididymides - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 343 S Microscopic 

prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

344 S Macroscopic 

all tissues - within normal limits 

344 S Microscopic 

epididymides - within normal limits prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 

345 S Macroscopic 

all tissues - within normal limits 

345 S Microscopic 

epididymides - within normal limits prostate gland - infiltration, lymphocytic, minimal seminal vesicles - within normal limits testes - within normal limits 

346 S Macroscopic 

all tissues - within normal limits 

346 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - MALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 346 S Microscopic 

seminal vesicles - within normal limits testes - within normal limits 

347 S Macroscopic 

all tissues - within normal limits 

347 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 

348 S Macroscopic 

all tissues - within normal limits 

348 S Microscopic 

epididymides - within normal limits prostate gland - within normal limits seminal vesicles - within normal limits testes - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 349 S Macroscopic 

all tissues - within normal limits 

349 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

350 S Macroscopic 

all tissues - within normal limits 

350 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

351 S Macroscopic 

all tissues - within normal limits 

351 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

352 S Macroscopic 

all tissues - within normal limits 

352 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

353 S Macroscopic 

all tissues - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 353 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

354 S Macroscopic 

all tissues - within normal limits 

354 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

355 S Macroscopic 

all tissues - within normal limits 

355 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

356 S Macroscopic 

all tissues - within normal limits 

356 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

357 S Macroscopic 

all tissues - within normal limits 

357 S Microscopic 

ovaries - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 357 S Microscopic 

uterus - within normal limits 

358 S Macroscopic 

all tissues - within normal limits 

358 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

359 S Macroscopic 

all tissues - within normal limits 

359 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

360 S Macroscopic 

all tissues - within normal limits 

360 S Microscopic 

ovaries - within normal limits uterus - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 150 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 361 S Macroscopic 

stomach, nonglandular - focus/foci, red, focal, mild 

362 S Macroscopic 

all tissues - within normal limits 

363 S Macroscopic 

all tissues - within normal limits 

364 S Macroscopic 

all tissues - within normal limits 

365 S Macroscopic 

all tissues - within normal limits 

366 S Macroscopic 

all tissues - within normal limits 

367 S Macroscopic 

all tissues - within normal limits 

368 S Macroscopic 

all tissues - within normal limits 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 150 mg/kg/day369r S Macroscopic 

all tissues - within normal limits 

370 S Macroscopic 

all tissues - within normal limits 

371r S Macroscopic 

all tissues - within normal limits 

372 S Macroscopic 

all tissues - within normal limits 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 600 mg/kg/day373r S Macroscopic 

all tissues - within normal limits 

374 D Macroscopic 

stomach, glandular - focus/foci, red, mucosa, moderate 

375 D Macroscopic 

all tissues - within normal limits 

376 D Macroscopic 

kidneys - discoloration, mottled, bilateral, mild stomach, glandular - focus/foci, black, mucosa, mild thyroid gland - pale, bilateral, mild 

uterus with cervix - within normal limits 

incomplete delivery. this animal had 14 imp

non-viable fetus in utero. this fetus was ex

377 S Macroscopic 

all tissues - within normal limits 

378r D Macroscopic 

stomach, glandular - focus/foci, red, mucosa, minimal 

379 D Macroscopic 

all tissues - within normal limits 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 600 mg/kg/day380 D Macroscopic 

thymus gland - discoloration, red, moderate 

- small, moderate 

381r D Macroscopic 

thymus gland - discoloration, red, mild 

- small, mild 

382 E Macroscopic 

liver - lobulation, accentuated, multiple lobes, mod

383 S Macroscopic 

all tissues - within normal limits 

384 S Macroscopic 

all tissues - within normal limits 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 385 D Macroscopic 

stomach, glandular - focus/foci, black, multiple, mucosa, mild 

385 D Microscopic 

ovaries - within normal limits stomach, glandular - within normal limits uterus - within normal limits 

non-correlated macro observation - stomach, glandular - focus/foci, black 

386 D Macroscopic 

stomach, glandular - focus/foci, black, mucosa, mild 

386 D Microscopic 

ovaries - within normal limits 

stomach, glandular - edema, mild 

- erosion/ulcer, mild 

corresponds to macroscopic observation (st

focus/foci, black) 

uterus - within normal limits 

387 S Macroscopic 

all tissues - within normal limits 

387 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

388 D Macroscopic 

stomach, glandular - focus/foci, black, mucosa, mild 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day388 D Microscopic 

ovaries - within normal limits stomach, glandular - erosion/ulcer, mild 

corresponds to macroscopic observation (st

focus/foci, black) 

uterus - within normal limits 

389 S Macroscopic 

all tissues - within normal limits 

389 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

390 S Macroscopic 

liver - discoloration, pink, multifocal, multiple lobes

390 S Microscopic 

liver - infiltration, mononuclear cell, minimal 

- necrosis, focal, minimal 

corresponds to macroscopic observation (liv

- vacuolation, periportal, hepatocytes, modera

ovaries - within normal limits uterus - within normal limits 

391r D Macroscopic 

stomach, glandular - focus/foci, black, mucosa, mild 

foci located in glandular and pylorus portions 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day391 D Microscopic 

ovaries - within normal limits 

stomach, glandular - edema, minimal 

- erosion/ulcer, mild 

corresponds to macroscopic observation (st

focus/foci, black) 

uterus - within normal limits 

392r S Macroscopic 

all tissues - within normal limits 

392 S Microscopic 

ovaries - within normal limits uterus - within normal limits 

393 E Macroscopic 

stomach, pylorus - erosion/ulcer, moderate 

stomach contents red. 

393 E Microscopic 

ovaries - within normal limits stomach, glandular - within normal limits uterus - within normal limits 

non-correlated macro observation - stomach, pylorus - erosion/ulcer 

394 D Macroscopic 

all tissues - within normal limits 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
MPI Research Study Number 1271-001 

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

Individual P Animal Data Record: Pathology - FEMALE 

Terminal 

Group, 

Animal Number Fate Tissue Observations 2400 mg/kg/day394 D Microscopic 

ovaries - within normal limits uterus - within normal limits 

395r D Macroscopic 

all tissues - within normal limits 

395 D Microscopic 

ovaries - within normal limits uterus - within normal limits 

396 S Macroscopic 

animal/whole body - body fat depleted, mild 

corresponds to antemortem observation (thi

396 S Microscopic 

ovaries - within normal limits uterus - within normal limits non-correlated macro observation - animal/whole body - body fat depleted 

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study 

rReplacement animal 
1271-001 

Appendix L 

Individual P Premating/Mating 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 starting 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 358
MPI Research Study Number 1271-001 

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

Individual P Premating Estrous Cycling 

Group, Cycle Length Mean Cycle Number of

Animal (Days) Length Cycles Number Cycle 1 Cycle 2 Cycle 3 Cycle 4 (Days) (Count) 

NA - Not Applicable/Not Available 

e - Excluded from summary statistics 

r Replacement animal 

0 mg/kg/day

349 4 4 4 4 4.00 4 350 6 4 6 5.33 3 351 NA NA NA 

352 5 4 5 3 4.25 4 353 3 4 5 5 4.25 4 354 5 5 5 5.00 3 355 3 5 5 5 4.50 4 356 5 4 4 4 4.25 4 357 4 4 4 4 4.00 4 358 4 4 4 4.00 3 359 5 5 5 4 4.75 4 360 6 4 3 4.33 3 

150 mg/kg/day

361 4 5 4 4.33 3 362 4 4.00 1 363 5 10e 4 4.50 2 

364 4 8 4 5.33 3 365 4 4 8 5.33 3 366 4 4 4 4 4.00 4 367 4 4.00 1 368 4 4 4 4 4.00 4 

369r 8 8.00 1 

370 4 4 4 4 4.00 4 

371r 4 4.00 1 

372 4 6 5 5.00 3 
MPI Research Study Number 1271-001 

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

Individual P Premating Estrous Cycling 

Group, Cycle Length Mean Cycle Number of

Animal (Days) Length Cycles Number Cycle 1 Cycle 2 Cycle 3 Cycle 4 (Days) (Count) 

NA - Not Applicable/Not Available 

e - Excluded from summary statistics 

r Replacement animal 

600 mg/kg/day

373r 8 8.00 1 

374 NA NA NA 

375 9 5 7.00 2 376 7 4 4 5.00 3 377 12e 4 4.00 1 

378r 4 4 6 4.67 3 

379 3 4 9 5.33 3 380 9 5 7.00 2 

381r 4 5 4 4.33 3 

382 8 5 6.50 2 383 NA NA NA 

384 4 4 4.00 2 

2400 mg/kg/day

385 NA NA NA 

386 5 4 4 4.33 3 387 3 4 5 4.00 3 388 NA NA NA 

389 7 5 4 5.33 3 390 4 5 8 5.67 3 

391r 4 4 4 4.00 3 

392r 5 4 4.50 2 

393 5 5 4 4.67 3 394 5 5 8 6.00 3 

395r 5 5.00 1 

396 4 5 5 4.67 3 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Estrous Cycle Stages 

Group, 

Animal Day of Study Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

DI - Diestrus MET - Metestrus PRO - Proestrus EST - Estrus 

S - Sperm 

r Replacement animal 

0 mg/kg/day

349 DI DI DI EST DI DI DI EST DI DI DI EST DI DI 

350 DI EST PRO DI DI DI EST EST DI DI DI EST MET DI 351 PRO DI DI DI DI DI DI DI DI DI DI DI PRO DI 352 EST DI DI EST EST EST PRO DI DI EST DI DI DI MET 353 EST DI PRO EST DI DI DI EST DI DI DI EST EST DI 354 DI DI DI EST EST DI DI DI DI EST DI DI DI EST 355 DI EST PRO EST EST DI DI DI EST EST DI DI DI EST 356 DI EST DI DI DI DI EST DI DI DI EST DI DI DI 357 DI EST DI DI DI EST DI DI DI EST DI DI DI EST 358 DI DI EST DI DI DI EST DI DI DI EST DI DI DI 359 EST DI EST DI EST EST PRO DI DI EST EST DI DI DI 360 DI DI DI DI EST DI DI DI DI DI EST DI DI DI 

150 mg/kg/day

361 DI DI DI DI DI EST PRO DI DI EST DI DI DI EST 

362 DI DI PRO PRO DI DI DI DI DI DI DI DI DI PRO 363 EST DI DI DI DI EST DI DI DI DI DI DI DI DI 364 EST DI DI DI EST DI DI DI PRO DI DI DI EST PRO 365 DI PRO DI EST DI DI DI EST DI DI DI EST DI DI 366 DI PRO DI EST DI DI DI EST DI DI DI EST DI DI 367 DI DI DI DI MET DI DI DI DI DI DI DI DI DI 368 DI EST DI DI DI EST PRO DI DI EST DI DI DI EST 

369r DI PRO EST EST DI DI DI DI DI DI DI EST DI DI 

370 DI EST PRO DI DI EST PRO DI DI EST DI DI DI EST 

371r DI DI DI DI DI DI DI EST PRO DI DI EST DI DI 

372 DI DI DI EST EST DI DI DI EST DI DI DI DI EST 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Estrous Cycle Stages 

Group, 

Animal Day of Study Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 

DI - Diestrus PRO - Proestrus EST - Estrus NA - Not Applicable/Not Available 

S - Sperm 

r Replacement animal 

600 mg/kg/day

373r DI EST DI DI DI DI DI DI PRO EST DI DI DI DI 

374 DI DI DI DI DI DI DI DI DI DI DI DI DI DI 

375 DI PRO EST EST DI DI DI DI DI DI DI EST EST DI 376 DI PRO EST DI DI DI DI DI DI EST DI DI DI EST 377 EST EST PRO EST DI DI DI DI DI DI DI DI EST EST 

378r EST DI DI DI EST DI DI DI EST DI DI DI DI EST 

379 DI EST PRO EST EST DI DI DI EST DI DI DI DI DI 380 DI DI EST EST DI DI DI DI DI DI DI EST EST DI 

381r DI EST DI DI DI EST DI DI DI DI EST DI DI DI 

382 DI DI DI DI EST DI EST DI DI DI DI DI EST DI 383 DI DI DI DI DI DI DI DI DI DI DI DI DI DI 

384 DI DI DI EST EST DI DI DI EST DI DI DI EST DI 

2400 mg/kg/day

385 DI DI EST EST DI DI DI DI DI DI DI DI DI PRO 

386 DI DI DI EST EST PRO DI DI DI EST DI DI DI EST 387 EST DI DI EST DI DI EST EST DI DI DI EST EST DI 388 DI DI DI DI DI DI DI DI DI DI DI DI DI DI 

389 EST PRO EST DI DI DI PRO EST DI DI DI DI EST DI 390 DI DI EST DI DI DI EST DI DI DI DI EST DI DI 

391r EST EST PRO DI DI EST DI DI DI EST DI DI DI EST 

392r DI NA PRO EST DI DI DI DI EST PRO DI DI EST DI 

393 DI DI EST EST DI PRO DI PRO EST DI DI DI EST EST 394 EST EST DI DI DI DI EST DI DI DI DI EST DI DI 

395r DI NA DI DI DI DI EST DI DI DI DI EST DI DI 

396 DI DI PRO EST EST DI DI DI EST DI DI DI DI EST 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Estrous Cycle Stages 

Group, 

Animal Day of Study Number 18 19 20 21 22 23 24 25 26 27 28 29 30 31 

DI - Diestrus MET - Metestrus PRO - Proestrus EST - Estrus 

S - Sperm P - Plug 

r Replacement animal 

0 mg/kg/day

349 DI DI EST DI PRO DI EST S 350 EST DI DI PRO S 351 DI DI DI DI S 352 EST PRO EST DI DI DI DI DI DI DI DI DI DI DI 

353 EST DI DI DI EST S 354 DI DI EST DI PRO EST S 355 DI EST EST DI DI PRO S 356 DI EST DI DI PRO EST S 357 EST DI DI EST S 358 DI PRO DI DI DI S 359 DI PRO EST S 360 EST MET DI DI EST S 

150 mg/kg/day

361 DI EST DI DI PRO S 362 DI PRO EST S 363 DI DI EST DI DI DI EST PRO DI DI DI S 

364 DI DI DI S 365 DI DI EST DI DI DI S 366 DI DI EST DI DI DI S 367 DI EST DI DI EST EST S 368 EST DI DI EST S 

369r 

370 EST DI DI DI P 

371r 

372 DI EST EST DI DI DI DI DI DI DI DI DI DI DI 
MPI Research Study Number 1271-001 

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

Individual P Premating/Mating Estrous Cycle Stages 

Group, 

Animal Day of Study Number 18 19 20 21 22 23 24 25 26 27 28 29 30 31 

DI - Diestrus MET - Metestrus PRO - Proestrus EST - Estrus 

S - Sperm P - Plug 

r Replacement animal 

600 mg/kg/day

373r 

374 DI EST DI DI DI S 375 EST DI DI DI S 376 EST DI DI DI S 377 EST DI DI DI EST EST S 

378r DI S 

379 EST DI DI DI DI DI EST DI DI DI DI DI DI DI 

380 EST DI DI DI DI DI DI DI DI DI DI DI DI DI 

381r DI EST DI DI EST S 

382 EST DI DI EST S 383 DI DI EST DI DI DI S 384 DI DI DI DI S 

2400 mg/kg/day

385 DI DI DI DI DI DI DI DI DI DI DI DI DI DI 

386 EST DI DI DI P 387 PRO DI DI EST S 388 DI DI DI DI PRO S 389 MET DI DI PRO S 390 DI EST EST DI DI DI S 

391r S 

392r S 

393 EST DI DI EST P 394 DI DI EST DI DI DI DI DI DI DI DI DI DI DI 

395r 

396 DI EST DI DI DI EST S 
1271-001 

Appendix M 

Individual P Mating Record 

Page 237 of 358
MPI Research Study Number 1271-001 

An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CA # 84-65-1) in Rats 

Individual Mating Record 

Group, 

Female Number Male Number Outcome Date of Cohabitation Date of Ins

NRMP - Normal pregnancy NA - Not Applicable/N

NP - Not pregnant r Replacement anim

0 mg/kg/day

349 303 NRMP 17JUL2006 22JU350 312 NRMP 17JUL2006 19JU351 301 NRMP 17JUL2006 19JU352 305 NP 17JUL2006 N353 310 NRMP 17JUL2006 20JU354 304 NRMP 17JUL2006 21JU355 307 NRMP 17JUL2006 21JU356 309 NRMP 17JUL2006 21JU357 308 NRMP 17JUL2006 19JU358 306 NRMP 17JUL2006 20JU359 311 NRMP 17JUL2006 18JU360 302 NRMP 17JUL2006 20JU

150 mg/kg/day

361 320 NRMP 17JUL2006 20JU362 315 NRMP 17JUL2006 18JU363 322 NRMP 17JUL2006 26JU364 314 NRMP 17JUL2006 18JU365 318 NRMP 17JUL2006 21JU366 319 NRMP 17JUL2006 21JU367 317 NRMP 17JUL2006 21JU

368 313 NRMP 17JUL2006 19JU

369r 321 NRMP 17JUL2006 18JU

370 316 NRMP 17JUL2006 19JU

371r 323 NP 17JUL2006 19JU

372 324r NP 17JUL2006 N
MPI Research Study Number 1271-001 

An Oral Reproduction/Developmental Toxicity Screening Study of Anthraquinone (CA # 84-65-1) in Rats 

Individual Mating Record 

Group, 

Female Number Male Number Outcome Date of Cohabitation Date of Ins

ALR - All resorptions NRMP - Normal pregnancy NA - Not Applicable/N

DNP - Died not pregnant PIDE - Pregnant incomplete delivery r Replacement anim

DP - Died pregnant PEM - Pregnant euthanized moribund 

600 mg/kg/day

373r 326 NRMP 17JUL2006 18JU

374 325 PIDE 17JUL2006 20JU375 328 NRMP 17JUL2006 19JU376 334 PIDE 17JUL2006 19JU377 336 NRMP 17JUL2006 21JU

378r 331 DP 17JUL2006 21JU

379 330 DP 17JUL2006 N380 335 PNMD 17JUL2006 N

381r 333r NRMP 17JUL2006 23JU

382 327 NRMP 17JUL2006 19JU383 332 NRMP 17JUL2006 21JU384 329 NRMP 17JUL2006 19JU

2400 mg/kg/day

385 348 DNP 17JUL2006 N386 338 DP 17JUL2006 19JU387 347 NRMP 17JUL2006 19JU388 346 DP 17JUL2006 20JU389 344 NRMP 17JUL2006 19JU390 341 NRMP 17JUL2006 21JU

391r 342 DP 17JUL2006 19JU

392r 343 NRMP 17JUL2006 19JU

393 339 PEM 17JUL2006 19JU394 340 DP 17JUL2006 N

395r 345 DP 17JUL2006 18JU

396 337r ALR 17JUL2006 21JU
1271-001 

Appendix N 

Individual P Natural Delivery and Litter Data 

Page 240 of 358
MPI Research Study Number 1271-001 

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

Individual P Natural Delivery and Litter Data 

____________________________________________________________________________________________________________

rReplacement animal 

Group, 

Female 

Gestation 

Length Number of Pups Delivered on Day 0 Implant 

Number (Days) Liveborn Stillborn Uncertain Total Scars 0 mg/kg/day

349 22 13 0 0 13 16 350 22 16 0 0 16 16 351 22 17 0 0 17 18 353 22 16 0 0 16 16 354 22 16 0 0 16 16 355 23 13 0 0 13 14 356 22 17 0 0 17 17 357 22 13 0 0 13 15 358 22 12 0 0 12 14 359 22 12 0 0 12 13 360 22 16 0 0 16 16 

150 mg/kg/day

361 23 13 0 0 13 14 362 22 10 0 0 10 11 363 22 12 1 0 13 14 364 23 13 0 0 13 15 365 23 14 0 0 14 15 366 23 12 0 0 12 14 367 23 14 0 0 14 14 368 22 10 0 0 10 13 369r 23 15 0 0 15 15 370 22 13 0 0 13 14 
MPI Research Study Number 1271-001 

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

Individual P Natural Delivery and Litter Data 

____________________________________________________________________________________________________________

PIDE - Pregnant incomplete delivery rReplacement animal x - Excluded from PNMD - Pregnant no mating date NA - Not Applicable/Not Available 

Group, 

Female 

Gestation 

Length Number of Pups Delivered on Day 0 Impla

Number (Days) Liveborn Stillborn Uncertain Total Scar600 mg/kg/day

373r 24 1 0 0 1 11 374 x PIDE 23 3 0 0 3 NA 375 24 6 5 0 11 13 376 x PIDE 23 0 14 0 14 14 377 23 11 0 0 11 11 380 PNMD NA 16 0 0 16 17 381r 23 6 0 0 6 8 382 23 0 13 0 13 15 383 22 12 2 0 14 14 384 23 13 0 0 13 15 

2400 mg/kg/day

387 23 1 2 0 3 10 389 23 12 0 0 12 13 390 22 13 0 0 13 15 392r 24 5 3 0 8 12 
MPI Research Study Number 1271-001 

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

Individual P Litter Size

Group, Number of Live Pups

Female Day 0 Day 4 Number M F M F 

r Replacement animal 

0 mg/kg/day 

349 4 9 4 9 

350 7 9 7 9 351 12 5 12 5 353 9 7 8 5 354 8 8 7 8 355 8 5 8 5 356 11 6 11 5 357 4 9 4 9 358 7 5 7 5 359 5 7 5 7 360 14 2 14 2 

150 mg/kg/day

361 6 7 0 0 

362 3 7 3 7 363 5 7 5 7 364 10 3 10 3 365 4 10 4 10 366 5 7 5 7 367 7 7 7 7 368 5 5 5 4 

369r 7 8 7 8 

370 5 8 5 8 
MPI Research Study Number 1271-001 

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

Individual P Litter Size

Group, Number of Live Pups

Female Day 0 Day 4 Number M F M F 

PIDE - Pregnant incomplete delivery r Replacement animal x - Excluded from sum

PNMD - Pregnant no mating date 

600 mg/kg/day

373r 0 0 0 0 

374 x PIDE 1 2 0 0 375 0 0 0 0 376 x PIDE 0 0 0 0 377 7 3 0 0 380 PNMD 11 5 0 0 

381r 0 6 0 0 

382 0 0 0 0 383 4 8 2 7 384 6 7 0 1 

2400 mg/kg/day

387 0 1 0 0 389 3 9 3 9 390 10 3 0 0 

392r 0 0 0 0 
1271-001 

Appendix O 

Individual F1 Pup Clinical Findings 

Page 245 of 358
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

34901 No Abnormalities Detected 0, 4 

34902 No Abnormalities Detected 0, 4 34903 No Abnormalities Detected 0, 4 34904 No Abnormalities Detected 0, 4 

34905 No Abnormalities Detected 0, 4 

34906 No Abnormalities Detected 0, 4 34907 No Abnormalities Detected 0, 4 

34908 No Abnormalities Detected 0, 4 34909 No Abnormalities Detected 0, 4 

34910 No Abnormalities Detected 0, 4 34911 No Abnormalities Detected 0, 4 

34912 No Abnormalities Detected 0, 4 

34913 No Abnormalities Detected 0, 4 35001 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35002 No Abnormalities Detected 0, 4 

35003 No Abnormalities Detected 0, 4 35004 No Abnormalities Detected 0, 4 35005 No Abnormalities Detected 0, 4 

35006 No Abnormalities Detected 0, 4 

35007 No Abnormalities Detected 0, 4 35008 No Abnormalities Detected 0, 4 

35009 No Abnormalities Detected 0, 4 35010 No Abnormalities Detected 0, 4 

35011 No Abnormalities Detected 0, 4 35012 No Abnormalities Detected 0, 4 

35013 No Abnormalities Detected 0, 4 

35014 No Abnormalities Detected 0, 4 35015 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35016 No Abnormalities Detected 0, 4 

35101 No Abnormalities Detected 0, 4 35102 No Abnormalities Detected 0, 4 35103 No Abnormalities Detected 0, 4 

35104 No Abnormalities Detected 0, 4 

35105 No Abnormalities Detected 0, 4 35106 No Abnormalities Detected 0, 4 

35107 No Abnormalities Detected 0, 4 35108 No Abnormalities Detected 0, 4 

35109 No Abnormalities Detected 0, 4 35110 No Abnormalities Detected 0, 4 

35111 No Abnormalities Detected 0, 4 

35112 No Abnormalities Detected 0, 4 35113 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35114 No Abnormalities Detected 0, 4 

35115 No Abnormalities Detected 0, 4 35116 No Abnormalities Detected 0, 4 35117 No Abnormalities Detected 0, 4 

35301 No Abnormalities Detected 0, 4 

35302 No Abnormalities Detected 0, 4 35303 No Abnormalities Detected 0, 4 

35304 No Abnormalities Detected 0, 4 35305 No Abnormalities Detected 0, 4 

35306 No Abnormalities Detected 0, 4 35307 No Abnormalities Detected 0, 4 

35308 No Abnormalities Detected 0, 4 

35309 No Abnormalities Detected 0 35310 No Abnormalities Detected 0 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35311 No Abnormalities Detected 0 

35312 No Abnormalities Detected 0, 4 35313 No Abnormalities Detected 0, 4 35314 No Abnormalities Detected 0, 4 

35315 No Abnormalities Detected 0, 4 

35316 No Abnormalities Detected 0, 4 35401 No Abnormalities Detected 0, 4 

35402 No Abnormalities Detected 0, 4 35403 No Abnormalities Detected 0, 4 

35404 No Abnormalities Detected 0, 4 35405 No Abnormalities Detected 0, 4 

35406 No Abnormalities Detected 0, 4 

35407 No Abnormalities Detected 0, 4 35409 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35410 No Abnormalities Detected 0, 4 

35411 No Abnormalities Detected 0, 4 35412 No Abnormalities Detected 0, 4 35413 No Abnormalities Detected 0, 4 

35414 No Abnormalities Detected 0, 4 

35415 No Abnormalities Detected 0, 4 35416 No Abnormalities Detected 0, 4 

35501 No Abnormalities Detected 0, 4 35502 No Abnormalities Detected 0, 4 

35503 No Abnormalities Detected 0, 4 35504 No Abnormalities Detected 0, 4 

35505 No Abnormalities Detected 0, 4 

35506 No Abnormalities Detected 0, 4 35507 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35508 No Abnormalities Detected 0, 4 

35509 No Abnormalities Detected 0, 4 35510 No Abnormalities Detected 0, 4 35511 No Abnormalities Detected 0, 4 

35512 No Abnormalities Detected 0, 4 

35513 No Abnormalities Detected 0, 4 35601 No Abnormalities Detected 0, 4 

35602 No Abnormalities Detected 0, 4 35603 No Abnormalities Detected 0, 4 

35604 No Abnormalities Detected 0, 4 35605 No Abnormalities Detected 0, 4 

35606 No Abnormalities Detected 0, 4 

35607 No Abnormalities Detected 0, 4 35608 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35609 No Abnormalities Detected 0, 4 

35610 No Abnormalities Detected 0, 4 35611 No Abnormalities Detected 0, 4 35612 No Abnormalities Detected 0, 4 

35613 No Abnormalities Detected 0, 4 

35614 No Abnormalities Detected 0, 4 35615 No Abnormalities Detected 0 

35616 No Abnormalities Detected 0, 4 35617 No Abnormalities Detected 0, 4 

35701 No Abnormalities Detected 0, 4 35702 No Abnormalities Detected 0, 4 

35703 No Abnormalities Detected 0, 4 

35704 No Abnormalities Detected 0, 4 35705 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35706 No Abnormalities Detected 0, 4 

35707 No Abnormalities Detected 0, 4 35708 No Abnormalities Detected 0, 4 35709 No Abnormalities Detected 0, 4 

35710 No Abnormalities Detected 0, 4 

35711 No Abnormalities Detected 0, 4 35712 No Abnormalities Detected 0, 4 

35713 No Abnormalities Detected 0, 4 35801 No Abnormalities Detected 0, 4 

35802 No Abnormalities Detected 0, 4 35803 No Abnormalities Detected 0, 4 

35804 No Abnormalities Detected 0, 4 

35805 No Abnormalities Detected 0, 4 35806 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35807 No Abnormalities Detected 0, 4 

35808 No Abnormalities Detected 0, 4 35809 No Abnormalities Detected 0, 4 35810 No Abnormalities Detected 0, 4 

35811 No Abnormalities Detected 0, 4 

35812 No Abnormalities Detected 0, 4 35901 No Abnormalities Detected 0, 4 

35902 No Abnormalities Detected 0, 4 35903 No Abnormalities Detected 0, 4 

35904 No Abnormalities Detected 0, 4 35905 No Abnormalities Detected 0, 4 

35906 No Abnormalities Detected 0, 4 

35907 No Abnormalities Detected 0, 4 35908 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

35909 No Abnormalities Detected 0, 4 

35910 No Abnormalities Detected 0, 4 35911 No Abnormalities Detected 0, 4 35912 No Abnormalities Detected 0, 4 

36001 No Abnormalities Detected 0, 4 

36002 No Abnormalities Detected 0, 4 36003 No Abnormalities Detected 0, 4 

36004 No Abnormalities Detected 0, 4 36005 No Abnormalities Detected 0, 4 

36006 No Abnormalities Detected 0, 4 36007 No Abnormalities Detected 0, 4 

36008 No Abnormalities Detected 0, 4 

36009 No Abnormalities Detected 0, 4 36010 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 0 mg/kg/day

36011 No Abnormalities Detected 0, 4 

36012 No Abnormalities Detected 0, 4 36013 No Abnormalities Detected 0, 4 36014 No Abnormalities Detected 0, 4 

36015 No Abnormalities Detected 0, 4 

36016 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

36101 No Abnormalities Detected 0 

36102 No Abnormalities Detected 0 36103 No Abnormalities Detected 0 

36104 No Abnormalities Detected 0 

36105 No Abnormalities Detected 0 36106 No Abnormalities Detected 0 

36107 No Abnormalities Detected 0 

36108 No Abnormalities Detected 0 36109 No Abnormalities Detected 0 36110 No Abnormalities Detected 0 

36111 No Abnormalities Detected 0 

36112 No Abnormalities Detected 0 36113 No Abnormalities Detected 0 

36201 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

36202 No Abnormalities Detected 0, 4 

36203 No Abnormalities Detected 0, 4 36204 No Abnormalities Detected 0 

Abrasion(s), Cranial region 4 Skin cold to touch 4 Skin discolored, Pale, Entire body 4 

36205 No Abnormalities Detected 0, 4 

36206 No Abnormalities Detected 0, 4 36207 No Abnormalities Detected 0, 4 36208 No Abnormalities Detected 0, 4 

36209 No Abnormalities Detected 0, 4 

36210 No Abnormalities Detected 0, 4 36301 No Abnormalities Detected 0, 4 

36302 No Abnormalities Detected 0, 4 36303 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

36304 No Abnormalities Detected 0, 4 

36305 No Abnormalities Detected 0, 4 36306 No Abnormalities Detected 0, 4 

36307 No Abnormalities Detected 0, 4 

36308 No Abnormalities Detected 0, 4 36309 No Abnormalities Detected 0, 4 

36310 No Abnormalities Detected 0, 4 

36311 No Abnormalities Detected 0, 4 36312 No Abnormalities Detected 0, 4 36401 No Abnormalities Detected 0, 4 

36402 No Abnormalities Detected 0, 4 

36403 No Abnormalities Detected 0, 4 36404 No Abnormalities Detected 0, 4 

36405 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

36406 No Abnormalities Detected 0, 4 

36407 No Abnormalities Detected 0, 4 36408 No Abnormalities Detected 0, 4 

36409 No Abnormalities Detected 0, 4 

36410 No Abnormalities Detected 0, 4 36411 No Abnormalities Detected 0, 4 

36412 No Abnormalities Detected 0, 4 

36413 No Abnormalities Detected 0, 4 36501 No Abnormalities Detected 0, 4 36502 No Abnormalities Detected 0, 4 

36503 No Abnormalities Detected 0, 4 

36504 No Abnormalities Detected 0, 4 36505 No Abnormalities Detected 0, 4 

36506 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

36507 No Abnormalities Detected 0, 4 

36508 No Abnormalities Detected 0, 4 36509 No Abnormalities Detected 0, 4 

36510 No Abnormalities Detected 0, 4 

36511 No Abnormalities Detected 0, 4 36512 No Abnormalities Detected 0, 4 

36513 No Abnormalities Detected 0, 4 

36514 No Abnormalities Detected 0, 4 36601 No Abnormalities Detected 0, 4 36602 No Abnormalities Detected 0, 4 

36603 No Abnormalities Detected 0, 4 

36604 No Abnormalities Detected 0, 4 36605 No Abnormalities Detected 0, 4 

36606 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

36607 No Abnormalities Detected 0, 4 

36608 No Abnormalities Detected 0, 4 36609 No Abnormalities Detected 0, 4 

36610 No Abnormalities Detected 0, 4 

36611 No Abnormalities Detected 0, 4 36612 No Abnormalities Detected 0, 4 

36701 No Abnormalities Detected 0, 4 

36702 No Abnormalities Detected 0, 4 36703 No Abnormalities Detected 0, 4 36704 No Abnormalities Detected 0, 4 

36705 No Abnormalities Detected 0, 4 

36706 No Abnormalities Detected 0, 4 36707 No Abnormalities Detected 0, 4 

36708 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

36709 No Abnormalities Detected 0, 4 

36710 No Abnormalities Detected 0, 4 36711 No Abnormalities Detected 0, 4 

36712 No Abnormalities Detected 0, 4 

36713 No Abnormalities Detected 0, 4 36714 No Abnormalities Detected 0, 4 

36801 No Abnormalities Detected 0, 4 

36802 No Abnormalities Detected 0, 4 36803 No Abnormalities Detected 0, 4 36804 No Abnormalities Detected 0, 4 

36805 No Abnormalities Detected 4 

Limb function impaired, Hind limb/left 0 Limb function impaired, Hind limb/right 0 

36806 No Abnormalities Detected 0, 4 36807 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 

r Replacement animal 

150 mg/kg/day

36808 No Abnormalities Detected 0 

36809 No Abnormalities Detected 4 

Limb function impaired, Hind limb/left 0 Limb function impaired, Hind limb/right 0 

36810 No Abnormalities Detected 0, 4 36901 r No Abnormalities Detected 0, 4 

36902 r No Abnormalities Detected 0, 4 36903 r No Abnormalities Detected 0, 4 36904 r No Abnormalities Detected 0, 4 36905 r No Abnormalities Detected 0, 4 36906 r No Abnormalities Detected 0, 4 36907 r No Abnormalities Detected 0, 4 36908 r No Abnormalities Detected 0, 4 36909 r No Abnormalities Detected 0, 4 36910 r No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 

r Replacement animal 

150 mg/kg/day

36911 r No Abnormalities Detected 0, 4 

36912 r No Abnormalities Detected 0, 4 36913 r No Abnormalities Detected 0, 4 36914 r No Abnormalities Detected 0, 4 36915 r No Abnormalities Detected 0, 4 37001 No Abnormalities Detected 0, 4 

37002 No Abnormalities Detected 0, 4 37003 No Abnormalities Detected 0, 4 37004 No Abnormalities Detected 0, 4 37005 No Abnormalities Detected 0, 4 37006 No Abnormalities Detected 0, 4 37007 No Abnormalities Detected 0, 4 37008 No Abnormalities Detected 0, 4 37009 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 150 mg/kg/day

37010 No Abnormalities Detected 0, 4 

37011 No Abnormalities Detected 0, 4 37012 No Abnormalities Detected 0, 4 

37013 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 600 mg/kg/day

37401 Skin cold to touch 0 

37402 No Abnormalities Detected 0 37701 Skin cold to touch 0 

37702 Skin cold to touch 0 

37703 Skin cold to touch 0 37704 Skin cold to touch 0 

37705 Skin cold to touch 0 

37706 Skin cold to touch 0 37707 Skin cold to touch 0 37708 Skin cold to touch 0 

37709 Skin cold to touch 0 

37710 Skin cold to touch 0 38001 Skin cold to touch 0 

38002 Skin cold to touch 0 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 600 mg/kg/day

38003 Activity decreased 0 

Skin cold to touch 0 

Skin discolored, Pale, Entire body 0 

38004 Skin cold to touch 0 

38005 Skin cold to touch 0 38006 Skin cold to touch 0 

38007 Skin cold to touch 0 

38008 Skin cold to touch 0 38009 Skin cold to touch 0 38010 Skin cold to touch 0 

38011 Skin cold to touch 0 

38012 Skin cold to touch 0 38013 Skin cold to touch 0 

38014 Skin cold to touch 0 38015 Skin cold to touch 0 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 

r Replacement animal 

600 mg/kg/day

38016 Skin cold to touch 0 

38101 r Skin cold to touch 0 

38102 r Skin cold to touch 0 38103 r Skin cold to touch 0 38104 r Skin cold to touch 0 38105 r Skin cold to touch 0 38106 r Skin cold to touch 0 38301 No Abnormalities Detected 0, 4 

38302 No Abnormalities Detected 0, 4 38303 No Abnormalities Detected 0 38304 No Abnormalities Detected 0 38305 No Abnormalities Detected 0, 4 38306 No Abnormalities Detected 0, 4 38307 No Abnormalities Detected 0, 4 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 600 mg/kg/day

38308 No Abnormalities Detected 0 

38309 No Abnormalities Detected 0, 4 38310 No Abnormalities Detected 0, 4 

38311 No Abnormalities Detected 0, 4 

38312 No Abnormalities Detected 0, 4 38313 No Abnormalities Detected 0 

38401 No Abnormalities Detected 0 

38402 No Abnormalities Detected 0 38403 No Abnormalities Detected 0 38404 No Abnormalities Detected 0 

38405 No Abnormalities Detected 0 

38407 No Abnormalities Detected 0 38408 No Abnormalities Detected 0 

38409 No Abnormalities Detected 0 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 600 mg/kg/day

38410 No Abnormalities Detected 0 

38411 No Abnormalities Detected 0 38412 No Abnormalities Detected 0 

Thin 4 

38413 No Abnormalities Detected 0 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 2400 mg/kg/day

38901 No Abnormalities Detected 0, 4 

38902 No Abnormalities Detected 0, 4 38903 No Abnormalities Detected 0, 4 38904 No Abnormalities Detected 0, 4 

38905 No Abnormalities Detected 0, 4 

38906 No Abnormalities Detected 0, 4 38907 No Abnormalities Detected 0, 4 

38908 No Abnormalities Detected 0, 4 

38909 No Abnormalities Detected 0, 4 38910 No Abnormalities Detected 0, 4 38911 No Abnormalities Detected 0, 4 

38912 No Abnormalities Detected 0, 4 

39001 No Abnormalities Detected 0 39002 No Abnormalities Detected 0 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Clinical Findings 

Group, 

Animal Number Lactation Day Sign Present 2400 mg/kg/day

39003 No Abnormalities Detected 0 

39004 No Abnormalities Detected 0 39005 No Abnormalities Detected 0 39006 No Abnormalities Detected 0 

39007 No Abnormalities Detected 0 

39008 No Abnormalities Detected 0 39009 No Abnormalities Detected 0 

39010 No Abnormalities Detected 0 

39011 No Abnormalities Detected 0 39012 No Abnormalities Detected 0 39013 No Abnormalities Detected 0 
1271-001 

Appendix P 

Individual F1 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 assigned (ascending numbers across the top). 

The dam number and the two-digit (e.g., 01) identification number comprise the pup number. 

Page 276 of 358
MPI Research Study Number 1271-001 

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

Individual F1 Pup Body Weight Values, g

Group, 

Female Pup # Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 

D - Died r Replacement animal

0 mg/kg/day 

349 7.2 6.8 7.2 6.8 6.3 6.3 6.9 7.0 7.0 6.4 6.4 7.2 6.3 

350 6.3 7.0 6.6 7.2 6.3 6.7 6.4 6.9 6.1 6.8 6.4 6.4 6.4 6.1 5.7 6.1 

351 8.0 7.6 6.5 7.3 8.2 6.2 6.9 7.0 6.4 7.0 6.8 7.4 7.3 6.5 6.7 6.2 6353 6.2 6.4 6.8 5.5 6.2 6.2 6.6 5.7 6.1 5.8 5.5 5.6 6.7 7.0 6.3 5.5 

354 6.2 6.3 5.3 6.0 5.9 6.4 6.0 D 5.8 5.9 5.7 5.6 5.6 5.5 5.4 5.4 

355 7.4 7.8 7.2 7.3 7.0 7.6 8.2 7.3 6.9 7.2 7.2 7.0 7.3 356 6.8 6.3 6.8 6.5 6.2 5.7 5.8 6.7 6.4 5.0 5.3 6.6 6.9 6.2 6.4 6.7 6357 7.0 7.2 6.6 7.3 5.4 6.6 6.5 6.7 6.4 6.7 5.9 8.8 6.5 358 7.1 7.0 6.8 7.1 7.2 6.6 5.8 6.2 6.9 6.7 6.5 6.1 359 6.5 6.3 6.6 7.0 7.3 6.0 6.2 6.7 5.5 6.3 6.1 6.1 360 7.8 7.4 6.7 7.8 7.2 7.0 6.9 6.8 7.1 7.1 6.4 7.3 7.0 6.9 6.7 6.8 

150 mg/kg/day

361 6.1 6.2 6.5 5.9 4.9 5.4 5.6 6.4 5.7 5.4 5.0 4.7 4.3 362 6.8 6.5 6.0 6.8 6.7 6.1 6.6 7.4 6.3 6.5 363 6.1 6.1 6.1 5.3 5.8 5.9 5.0 6.1 5.2 5.9 5.8 5.8 S 364 6.3 6.4 6.2 7.4 6.2 6.5 5.9 6.1 5.8 6.1 6.1 5.5 6.3 365 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 7.0 366 6.5 6.4 5.6 6.5 6.7 6.3 6.2 6.2 6.4 6.6 5.8 6.2 367 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 6.7 368 6.5 7.6 7.1 7.1 7.0 6.6 6.5 6.8 6.6 6.8 369 r 6.6 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 5.7 

370 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 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Body Weight Values, g

Group, 

Female Pup # Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 

D - Died r Replacement animal x - Excluded from sum

S - Stillborn PIDE - Pregnant incomplete delivery Y - Cannibalized 

600 mg/kg/day

373 r Y 

374 x PIDE 5.4 5.0 D 375 S S S S S D Y Y Y Y Y 376 x PIDE S S S S S S S S S S S S S S 377 4.1 4.2 4.4 4.7 4.7 4.2 4.5 4.4 4.6 4.2 Y 

380 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 3.5 381 r 7.4 7.0 7.2 6.6 6.2 6.7 

382 S S S S S S S S S S S S S 383 5.8 5.8 5.4 5.1 5.5 4.0 5.5 4.8 5.0 5.2 4.8 5.6 4.8 S 

384 6.0 5.9 5.9 5.3 6.4 D 5.5 5.0 4.7 5.3 5.8 5.1 3.9 

2400 mg/kg/day

387 S S D 

389 6.6 6.8 6.4 6.3 6.1 6.0 5.9 5.7 6.2 6.5 6.2 5.6 390 6.7 6.0 6.5 6.4 6.7 8.4 6.7 6.1 6.1 6.4 5.4 5.7 6.6 

392 r S S S D Y Y Y Y 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Body Weight Values, g

Group, 

Female Pup # Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 

D - Died r Replacement animal

Z - Missing from cage 

0 mg/kg/day

349 10.7 10.7 10.1 9.6 9.7 9.2 9.8 9.6 10.5 9.7 10.2 10.2 9.6 

350 9.5 10.2 9.9 9.4 8.2 9.4 9.8 9.4 8.5 8.9 9.7 9.1 8.7 8.6 9.1 8.9 

351 12.8 10.7 9.6 10.6 11.1 9.3 11.3 10.3 9.5 10.3 10.9 11.3 11.4 10.7 10.1 10.1 1

353 8.8 8.9 8.4 6.3 7.9 8.2 9.0 7.6 D Z D 6.3 7.6 8.1 7.9 6.5 

354 8.1 9.2 7.9 7.9 9.2 9.6 8.9 D 8.9 8.4 7.7 7.9 8.3 7.5 8.1 8.1 

355 9.6 11.3 10.0 9.9 11.1 9.5 11.4 10.0 10.8 9.3 9.1 9.9 10.7 356 10.2 8.8 9.1 9.5 7.9 8.5 8.4 9.9 8.6 8.2 10.1 9.7 9.5 7.8 D 10.2 

357 10.4 9.5 10.4 10.1 7.1 9.0 8.0 10.0 9.0 10.4 7.5 10.2 9.7 

358 9.6 11.4 10.8 10.3 11.1 10.0 8.8 9.1 9.6 9.9 9.9 9.1 359 10.3 9.4 9.9 9.6 10.3 9.0 9.1 9.6 9.5 9.6 9.0 10.1 360 11.0 10.9 9.5 10.7 11.0 9.7 9.9 10.0 9.8 9.5 9.0 10.5 10.5 10.6 10.1 10.2 

150 mg/kg/day

361 D D D D D D D D D D D D D 362 7.4 7.9 7.3 6.4 7.8 7.6 8.0 7.8 7.9 7.9 363 7.7 7.2 7.4 6.8 6.9 7.3 6.6 7.5 6.8 7.1 6.6 7.3 S 364 8.6 8.2 8.1 9.7 9.2 9.2 8.4 7.5 7.9 8.0 8.1 7.5 8.4 365 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 10.9 

366 9.4 7.9 8.2 7.3 9.2 8.8 7.3 7.4 8.5 8.9 7.9 8.4 367 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 9.4 368 8.9 9.6 10.2 9.9 7.7 9.4 9.2 D 8.9 10.0 369 r 9.4 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 6.4 

370 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 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Body Weight Values, g

Group, 

Female Pup # Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 

D - Died Z - Missing from cage x - Excluded from sum

S - Stillborn r Replacement animal 

E - Euthanized in extremis PIDE - Pregnant incomplete delivery Y - Cannibalized 

600 mg/kg/day

373 r Y 

374 X PIDE E E D 375 S S S S S D Y Y Y Y Y 376 X PIDE S S S S S S S S S S S S S S 377 Z D Z D D Z D D Z D Y 380 D D D D D D Y D D D D D D D D D 381 r E D D D E D 382 S S S S S S S S S S S S S 383 4.7 4.9 D D 4.4 3.3 4.1 S 4.0 4.5 3.9 4.2 D S 

384 D D D D D D D D D D D 3.8 D 

2400 mg/kg/day

387 S S D 

389 8.8 8.5 8.8 8.7 8.4 7.4 8.8 8.5 8.6 9.0 8.8 7.1 390 D Z D Z Z Z D D Z D D D D 392 r S S S D Y Y Y Y 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Sex and Status

Group, 

Female Pup # Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 

SEX CODES: M - Male F - Female PUP STATUS CODES: N - Scheduled necropsy D - Died Z - Missing from cage rReplacement animal 

0 mg/kg/day 

349 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 4 

350 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 4 351 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 353 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 4 354 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 4 355 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 4 356 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 357 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 4 358 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 359 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 360 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 4 

150 mg/kg/day

361 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 2 

362 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 363 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 0 364 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 4 365 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 4 366 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 367 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 4 368 MN 4 MN 4 MN 4 MN 4 MN 4 FN 4 FN 4 FD 2 FN 4 FN 4 

369r 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 

370 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 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Sex and Status

Group, 

Female Pup # Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 

SEX CODES: M - Male F - Female U - Unsexed PUP STATUS CODES: N - Scheduled necropsy Y - Cannibalized  S - Stillborn D - Died Z - Missing from cage E - Euthanized in extremis rReplacement animal 1Pregnant incomplete delivery 

600 mg/kg/day

373r UY 0 

3741 ME 0 FE 0 FD 0 

375 MS 0 MS 0 FS 0 FS 0 FS 0 UD 0 UY 0 UY 0 UY 0 UY 0 UY 0 

3761 MS 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 0 

377 MZ 1 MD 1 MZ 1 MD 1 MD 1 MZ 1 MD 1 FD 1 FZ 2 FD 1 UY 0 380 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 2 

381r FE 3 FD 3 FD 3 FD 3 FE 3 FD 3 

382 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 0 383 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 0 384 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 3 

2400 mg/kg/day

387 MS 0 FS 0 FD 0 

389 MN 4 MN 4 MN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 FN 4 390 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 2 

392r MS 0 MS 0 FS 0 UD 0 UY 0 UY 0 UY 0 UY 0 
1271-001 

Appendix Q 

Individual F1 Pup Animal Data Record: Pathology 

Page 283 of 358
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Stillborn Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study B – Stillborn U – Unable to Determine Sex NT – N

363-NT F B Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Stillborn Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study B – Stillborn U – Unable to Determine Sex NT – N

375-NT1 M B Macroscopic

external appearance - within normal limits 

375-NT2 M B Macroscopic

external appearance - within normal limits 

375-NT3 F B Macroscopic

external appearance - within normal limits 

375-NT4 F B Macroscopic

external appearance - within normal limits 

375-NT5 F B Macroscopic

external appearance - within normal limits 

376-NT1 M B Macroscopic

external appearance - within normal limits 

376-NT2 M B Macroscopic

external appearance - within normal limits 

376-NT3 M B Macroscopic

external appearance - within normal limits 

376-NT4 F B Macroscopic

external appearance - within normal limits 

376-NT5 F B Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Stillborn Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study B – Stillborn U – Unable to Determine Sex NT – N

382-NT1 M B Macroscopic

external appearance - within normal limits 

382-NT2 M B Macroscopic

external appearance - within normal limits 

382-NT3 M B Macroscopic

external appearance - within normal limits 

382-NT4 M B Macroscopic

external appearance - within normal limits 

382-NT5 M B Macroscopic

external appearance - within normal limits 

382-NT6 F B Macroscopic

external appearance - within normal limits 

382-NT7 F B Macroscopic

external appearance - within normal limits 

382-NT8 F B Macroscopic

external appearance - within normal limits 

382-NT9 F B Macroscopic

external appearance - within normal limits 

382-NT10 F B Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Stillborn Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study B – Stillborn U – Unable to Determine Sex NT – N

382-NT11 F B Macroscopic

external appearance - within normal limits 

382-NT12 F B Macroscopic

external appearance - within normal limits 

382-NT13 F B Macroscopic

external appearance - within normal limits 

383-NT1 F B Macroscopic

external appearance - within normal limits 

383-NT2 F B Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Stillborn Pups 

Group,

Animal Number Sex Fate Tissue Observations

2400 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study B – Stillborn U – Unable to Determine Sex NT – N

387-NT1 M B Macroscopic

external appearance - within normal limits 

387-NT2 F B Macroscopic

external appearance - within normal limits 

392-NT1 M B Macroscopic

external appearance - within normal limits 

392-NT2 M B Macroscopic

external appearance - within normal limits 

392-NT3 F B Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed353-09 M D Macroscopic

external appearance - within normal limits 

353-11 F D Macroscopic

external appearance - within normal limits 

354-NT M D Macroscopic

external appearance - within normal limits 

356-15 F D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

361-01 M D Macroscopic

external appearance - within normal limits 

361-02 M D Macroscopic

external appearance - within normal limits 

361-03 M D Macroscopic

external appearance - within normal limits 

361-04 M D Macroscopic

external appearance - within normal limits 

361-05 M D Macroscopic

external appearance - within normal limits 

361-06 M D Macroscopic

external appearance - within normal limits 

361-07 F D Macroscopic

external appearance - within normal limits 

361-08 F D Macroscopic

external appearance - within normal limits 

361-09 F D Macroscopic

external appearance - within normal limits 

361-10 F D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

361-11 F D Macroscopic

external appearance - within normal limits 

361-12 F D Macroscopic

external appearance - within normal limits 

361-13 F D Macroscopic

external appearance - within normal limits 

368-08 F D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

374-01 M E Macroscopic

external appearance - within normal limits 

374-02 F E Macroscopic

external appearance - within normal limits 

377-02 M D Macroscopic

external appearance - within normal limits 

377-04 M D Macroscopic

external appearance - within normal limits 

377-05 M D Macroscopic

external appearance - within normal limits 

377-07 M D Macroscopic

external appearance - within normal limits 

377-08 F D Macroscopic

external appearance - within normal limits 

377-10 F D Macroscopic

external appearance - within normal limits 

380-01 M D Macroscopic

external appearance - within normal limits 

380-02 M D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

380-03 M D Macroscopic

external appearance - within normal limits 

380-04 M D Macroscopic

external appearance - within normal limits 

380-05 M D Macroscopic

external appearance - within normal limits 

380-06 M D Macroscopic

external appearance - within normal limits 

380-08 M D Macroscopic

external appearance - within normal limits 

380-09 M D Macroscopic

external appearance - within normal limits 

380-10 M D Macroscopic

external appearance - within normal limits 

380-11 M D Macroscopic

external appearance - within normal limits 

380-12 F D Macroscopic

external appearance - within normal limits 

380-13 F D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

380-14 F D Macroscopic

external appearance - within normal limits 

380-15 F D Macroscopic

external appearance - within normal limits 

380-16 F D Macroscopic

external appearance - within normal limits 

381-01 F E Macroscopic

external appearance - within normal limits 

381-02 F D Macroscopic

external appearance - within normal limits 

381-03 F D Macroscopic

external appearance - within normal limits 

381-04 F D Macroscopic

external appearance - within normal limits 

381-05 F E Macroscopic

external appearance - within normal limits 

381-06 F D Macroscopic

external appearance - within normal limits 

383-03 M D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

383-04 M D Macroscopic

external appearance - within normal limits 

383-13 F D Macroscopic

external appearance - within normal limits 

384-NT M D Macroscopic

external appearance - within normal limits 

384-01 M D Macroscopic

external appearance - within normal limits 

384-02 M D Macroscopic

external appearance - within normal limits 

384-03 M D Macroscopic

external appearance - within normal limits 

384-04 M D Macroscopic

external appearance - within normal limits 

384-05 M D Macroscopic

external appearance - within normal limits 

384-06 F D Macroscopic

external appearance - within normal limits 

384-07 F D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

384-08 F D Macroscopic

external appearance - within normal limits 

384-09 F D Macroscopic

external appearance - within normal limits 

384-10 F D Macroscopic

external appearance - within normal limits 

384-13 F D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died On Study Pups 

Group,

Animal Number Sex Fate Tissue Observations

2400 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

387-NT3 F D Macroscopic

external appearance - within normal limits 

390-01 M D Macroscopic

external appearance - within normal limits 

390-03 M D Macroscopic

external appearance - within normal limits 

390-07 M D Macroscopic

external appearance - within normal limits 

390-08 M D Macroscopic

external appearance - within normal limits 

390-10 M D Macroscopic

external appearance - within normal limits 

390-11 F D Macroscopic

external appearance - within normal limits 

390-12 F D Macroscopic

external appearance - within normal limits 

390-13 F D Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died on Study- Unsexed Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed 

373-NT U D Macroscopic

external appearance - too cannibalized to examine 

375-NT6 U D Macroscopic

external appearance - too cannibalized to examine 

375-NT7 U D Macroscopic

external appearance - too cannibalized to examine 

375-NT8 U D Macroscopic

external appearance - too cannibalized to examine 

375-NT9 U D Macroscopic

external appearance - too cannibalized to examine 

375-NT10 U D Macroscopic

external appearance - too cannibalized to examine 

375-NT11 U D Macroscopic

external appearance - too cannibalized to examine 

376-NT6 U B Macroscopic

external appearance - within normal limits 

376-NT7 U B Macroscopic

external appearance - within normal limits 

376-NT8 U B Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died on Study- Unsexed Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed 

376-NT9 U B Macroscopic

external appearance - within normal limits 

376-NT10 U B Macroscopic

external appearance - within normal limits 

376-NT11 U B Macroscopic

external appearance - within normal limits 

376-NT12 U B Macroscopic

external appearance - within normal limits 

376-NT13 U B Macroscopic

external appearance - within normal limits 

376-NT14 U B Macroscopic

external appearance - within normal limits 

377-NT U D Macroscopic

external appearance - too cannibalized to examine 

380-07 U D Macroscopic

external appearance - too cannibalized to examine 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Died on Study- Unsexed Pups 

Group,

Animal Number Sex Fate Tissue Observations

2400 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed 

392-NT4 U D Macroscopic

external appearance - too cannibalized to examine 

392-NT5 U D Macroscopic

external appearance - too cannibalized to examine 

392-NT6 U D Macroscopic

external appearance - too cannibalized to examine 

392-NT7 U D Macroscopic

external appearance - too cannibalized to examine 

392-NT8 U D Macroscopic

external appearance - too cannibalized to examine 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed349-01 M S Macroscopic

external appearance - within normal limits 

349-02 M S Macroscopic

external appearance - within normal limits 

349-03 M S Macroscopic

external appearance - within normal limits 

349-04 F S Macroscopic

external appearance - within normal limits 

349-05 F S Macroscopic

external appearance - within normal limits 

349-06 F S Macroscopic

external appearance - within normal limits 

349-07 M S Macroscopic

external appearance - within normal limits 

349-08 F S Macroscopic

external appearance - within normal limits 

349-09 F S Macroscopic

external appearance - within normal limits 

349-10 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed349-11 F S Macroscopic

external appearance - within normal limits 

349-12 F S Macroscopic

external appearance - within normal limits 

349-13 F S Macroscopic

external appearance - within normal limits 

350-01 M S Macroscopic

external appearance - within normal limits 

350-02 M S Macroscopic

external appearance - within normal limits 

350-03 M S Macroscopic

external appearance - within normal limits 

350-04 M S Macroscopic

external appearance - within normal limits 

350-05 M S Macroscopic

external appearance - within normal limits 

350-06 M S Macroscopic

external appearance - within normal limits 

350-07 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed350-08 F S Macroscopic

external appearance - within normal limits 

350-09 F S Macroscopic

external appearance - within normal limits 

350-10 F S Macroscopic

external appearance - within normal limits 

350-11 F S Macroscopic

external appearance - within normal limits 

350-12 F S Macroscopic

external appearance - within normal limits 

350-13 F S Macroscopic

external appearance - within normal limits 

350-14 F S Macroscopic

external appearance - within normal limits 

350-15 F S Macroscopic

external appearance - within normal limits 

350-16 F S Macroscopic

external appearance - within normal limits 

351-01 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed351-02 M S Macroscopic

external appearance - within normal limits 

351-03 M S Macroscopic

external appearance - within normal limits 

351-04 M S Macroscopic

external appearance - within normal limits 

351-05 M S Macroscopic

external appearance - within normal limits 

351-06 M S Macroscopic

external appearance - within normal limits 

351-07 M S Macroscopic

external appearance - within normal limits 

351-08 M S Macroscopic

external appearance - within normal limits 

351-09 M S Macroscopic

external appearance - within normal limits 

351-10 M S Macroscopic

external appearance - within normal limits 

351-11 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed351-12 M S Macroscopic

external appearance - within normal limits 

351-13 F S Macroscopic

external appearance - within normal limits 

351-14 F S Macroscopic

external appearance - within normal limits 

351-15 F S Macroscopic

external appearance - within normal limits 

351-16 F S Macroscopic

external appearance - within normal limits 

351-17 F S Macroscopic

external appearance - within normal limits 

353-01 M S Macroscopic

external appearance - within normal limits 

353-02 M S Macroscopic

external appearance - within normal limits 

353-03 M S Macroscopic

external appearance - within normal limits 

353-04 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed353-05 M S Macroscopic

external appearance - within normal limits 

353-06 M S Macroscopic

external appearance - within normal limits 

353-07 M S Macroscopic

external appearance - within normal limits 

353-08 M S Macroscopic

external appearance - within normal limits 

353-12 F S Macroscopic

external appearance - within normal limits 

353-13 F S Macroscopic

external appearance - within normal limits 

353-14 F S Macroscopic

external appearance - within normal limits 

353-15 F S Macroscopic

external appearance - within normal limits 

353-16 F S Macroscopic

external appearance - within normal limits 

354-01 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed354-02 M S Macroscopic

external appearance - within normal limits 

354-03 M S Macroscopic

external appearance - within normal limits 

354-04 M S Macroscopic

external appearance - within normal limits 

354-05 M S Macroscopic

external appearance - within normal limits 

354-06 M S Macroscopic

external appearance - within normal limits 

354-07 M S Macroscopic

external appearance - within normal limits 

354-09 F S Macroscopic

external appearance - within normal limits 

354-10 F S Macroscopic

external appearance - within normal limits 

354-11 F S Macroscopic

external appearance - within normal limits 

354-12 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed354-13 F S Macroscopic

external appearance - within normal limits 

354-14 F S Macroscopic

external appearance - within normal limits 

354-15 F S Macroscopic

external appearance - within normal limits 

354-16 F S Macroscopic

external appearance - within normal limits 

355-01 M S Macroscopic

external appearance - within normal limits 

355-02 M S Macroscopic

external appearance - within normal limits 

355-03 M S Macroscopic

external appearance - within normal limits 

355-04 M S Macroscopic

external appearance - within normal limits 

355-05 M S Macroscopic

external appearance - within normal limits 

355-06 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed355-07 M S Macroscopic

external appearance - within normal limits 

355-08 M S Macroscopic

external appearance - within normal limits 

355-09 F S Macroscopic

external appearance - within normal limits 

355-10 F S Macroscopic

external appearance - within normal limits 

355-11 F S Macroscopic

external appearance - within normal limits 

355-12 F S Macroscopic

external appearance - within normal limits 

355-13 F S Macroscopic

external appearance - within normal limits 

356-01 M S Macroscopic

external appearance - within normal limits 

356-02 M S Macroscopic

external appearance - within normal limits 

356-03 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed356-04 M S Macroscopic

external appearance - within normal limits 

356-05 M S Macroscopic

external appearance - within normal limits 

356-06 M S Macroscopic

external appearance - within normal limits 

356-07 M S Macroscopic

external appearance - within normal limits 

356-08 M S Macroscopic

external appearance - within normal limits 

356-09 M S Macroscopic

external appearance - within normal limits 

356-10 M S Macroscopic

external appearance - within normal limits 

356-11 M S Macroscopic

external appearance - within normal limits 

356-12 F S Macroscopic

external appearance - within normal limits 

356-13 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed356-14 F S Macroscopic

external appearance - within normal limits 

356-16 F S Macroscopic

external appearance - within normal limits 

356-17 F S Macroscopic

external appearance - within normal limits 

357-01 M S Macroscopic

external appearance - within normal limits 

357-02 M S Macroscopic

external appearance - within normal limits 

357-03 M S Macroscopic

external appearance - within normal limits 

357-04 M S Macroscopic

external appearance - within normal limits 

357-05 F S Macroscopic

external appearance - within normal limits 

357-06 F S Macroscopic

external appearance - within normal limits 

357-07 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed357-08 F S Macroscopic

external appearance - within normal limits 

357-09 F S Macroscopic

external appearance - within normal limits 

357-10 F S Macroscopic

external appearance - within normal limits 

357-11 F S Macroscopic

external appearance - within normal limits 

357-12 F S Macroscopic

external appearance - within normal limits 

357-13 F S Macroscopic

external appearance - within normal limits 

358-01 M S Macroscopic

external appearance - within normal limits 

358-02 M S Macroscopic

external appearance - within normal limits 

358-03 M S Macroscopic

external appearance - within normal limits 

358-04 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed358-05 M S Macroscopic

external appearance - within normal limits 

358-06 M S Macroscopic

external appearance - within normal limits 

358-07 M S Macroscopic

external appearance - within normal limits 

358-09 F S Macroscopic

external appearance - within normal limits 

358-10 F S Macroscopic

external appearance - within normal limits 

358-11 F S Macroscopic

external appearance - within normal limits 

358-12 F S Macroscopic

external appearance - within normal limits 

358-13 F S Macroscopic

external appearance - within normal limits 

359-01 M S Macroscopic

external appearance - within normal limits 

359-02 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed359-03 M S Macroscopic

external appearance - within normal limits 

359-04 M S Macroscopic

external appearance - within normal limits 

359-05 M S Macroscopic

external appearance - within normal limits 

359-06 F S Macroscopic

external appearance - within normal limits 

359-07 F S Macroscopic

external appearance - within normal limits 

359-08 F S Macroscopic

external appearance - within normal limits 

359-09 F S Macroscopic

external appearance - within normal limits 

359-10 F S Macroscopic

external appearance - within normal limits 

359-11 F S Macroscopic

external appearance - within normal limits 

359-12 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

360-01 M S Macroscopic

external appearance - within normal limits 

360-02 M S Macroscopic

external appearance - within normal limits 

360-03 M S Macroscopic

external appearance - within normal limits 

360-04 M S Macroscopic

external appearance - within normal limits 

360-05 M S Macroscopic

external appearance - within normal limits 

360-06 M S Macroscopic

external appearance - within normal limits 

360-07 M S Macroscopic

external appearance - within normal limits 

360-08 M S Macroscopic

external appearance - within normal limits 

360-09 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

0 mg/kg/day S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed360-10 M S Macroscopic

external appearance - within normal limits 

360-11 M S Macroscopic

external appearance - within normal limits 

360-12 M S Macroscopic

external appearance - within normal limits 

360-13 M S Macroscopic

external appearance - within normal limits 

360-14 M S Macroscopic

external appearance - within normal limits 

360-15 F S Macroscopic

external appearance - within normal limits 

360-16 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

362-01 M S Macroscopic

external appearance - within normal limits 

362-02 M S Macroscopic

external appearance - within normal limits 

362-03 M S Macroscopic

external appearance - within normal limits 

362-04 F S Macroscopic

external appearance - within normal limits 

362-05 F S Macroscopic

external appearance - within normal limits 

362-06 F S Macroscopic

external appearance - within normal limits 

362-07 F S Macroscopic

external appearance - within normal limits 

362-08 F S Macroscopic

external appearance - within normal limits 

362-09 F S Macroscopic

external appearance - within normal limits 

362-10 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

363-01 M S Macroscopic

external appearance - within normal limits 

363-02 M S Macroscopic

external appearance - within normal limits 

363-03 M S Macroscopic

external appearance - within normal limits 

363-04 M S Macroscopic

external appearance - within normal limits 

363-05 M S Macroscopic

external appearance - within normal limits 

363-06 F S Macroscopic

external appearance - within normal limits 

363-07 F S Macroscopic

external appearance - within normal limits 

363-08 F S Macroscopic

external appearance - within normal limits 

363-09 F S Macroscopic

external appearance - within normal limits 

363-10 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

363-11 F S Macroscopic

external appearance - within normal limits 

363-12 F S Macroscopic

external appearance - within normal limits 

364-01 M S Macroscopic

external appearance - within normal limits 

364-02 M S Macroscopic

external appearance - within normal limits 

364-03 M S Macroscopic

external appearance - within normal limits 

364-04 M S Macroscopic

external appearance - within normal limits 

364-05 M S Macroscopic

external appearance - within normal limits 

364-06 M S Macroscopic

external appearance - within normal limits 

364-07 M S Macroscopic

external appearance - within normal limits 

364-08 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

364-09 M S Macroscopic

external appearance - within normal limits 

364-10 M S Macroscopic

external appearance - within normal limits 

364-11 F S Macroscopic

external appearance - within normal limits 

364-12 F S Macroscopic

external appearance - within normal limits 

364-13 F S Macroscopic

external appearance - within normal limits 

365-01 M S Macroscopic

external appearance - within normal limits 

365-02 M S Macroscopic

external appearance - within normal limits 

365-03 M S Macroscopic

external appearance - within normal limits 

365-04 M S Macroscopic

external appearance - within normal limits 

365-05 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

365-06 F S Macroscopic

external appearance - within normal limits 

365-07 F S Macroscopic

external appearance - within normal limits 

365-08 F S Macroscopic

external appearance - within normal limits 

365-09 F S Macroscopic

external appearance - within normal limits 

365-10 F S Macroscopic

external appearance - within normal limits 

365-11 F S Macroscopic

external appearance - within normal limits 

365-12 F S Macroscopic

external appearance - within normal limits 

365-13 F S Macroscopic

external appearance - within normal limits 

365-14 F S Macroscopic

external appearance - within normal limits 

366-01 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

366-02 M S Macroscopic

external appearance - within normal limits 

366-03 M S Macroscopic

external appearance - within normal limits 

366-04 M S Macroscopic

external appearance - within normal limits 

366-05 M S Macroscopic

external appearance - within normal limits 

366-06 F S Macroscopic

external appearance - within normal limits 

366-07 F S Macroscopic

external appearance - within normal limits 

366-08 F S Macroscopic

external appearance - within normal limits 

366-09 F S Macroscopic

external appearance - within normal limits 

366-10 F S Macroscopic

external appearance - within normal limits 

366-11 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

366-12 F S Macroscopic

external appearance - within normal limits 

367-01 M S Macroscopic

external appearance - within normal limits 

367-02 M S Macroscopic

external appearance - within normal limits 

367-03 M S Macroscopic

external appearance - within normal limits 

367-04 M S Macroscopic

external appearance - within normal limits 

367-05 M S Macroscopic

external appearance - within normal limits 

367-06 M S Macroscopic

external appearance - within normal limits 

367-07 M S Macroscopic

external appearance - within normal limits 

367-08 F S Macroscopic

external appearance - within normal limits 

367-09 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

367-10 F S Macroscopic

external appearance - within normal limits 

367-11 F S Macroscopic

external appearance - within normal limits 

367-12 F S Macroscopic

external appearance - within normal limits 

367-13 F S Macroscopic

external appearance - within normal limits 

367-14 F S Macroscopic

external appearance - within normal limits 

368-01 M S Macroscopic

external appearance - within normal limits 

368-02 M S Macroscopic

external appearance - within normal limits 

368-03 M S Macroscopic

external appearance - within normal limits 

368-04 M S Macroscopic

external appearance - within normal limits 

368-05 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

368-06 F S Macroscopic

external appearance - within normal limits 

368-07 F S Macroscopic

external appearance - within normal limits 

368-09 F S Macroscopic

external appearance - within normal limits 

368-10 F S Macroscopic

external appearance - within normal limits 

369-01 M S Macroscopic

external appearance - within normal limits 

369-02 M S Macroscopic

external appearance - within normal limits 

369-03 M S Macroscopic

external appearance - within normal limits 

369-04 M S Macroscopic

external appearance - within normal limits 

369-05 M S Macroscopic

external appearance - within normal limits 

369-06 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

369-07 M S Macroscopic

external appearance - within normal limits 

369-08 F S Macroscopic

external appearance - within normal limits 

369-09 F S Macroscopic

external appearance - within normal limits 

369-10 F S Macroscopic

external appearance - within normal limits 

369-11 F S Macroscopic

external appearance - within normal limits 

369-12 F S Macroscopic

external appearance - within normal limits 

369-13 F S Macroscopic

external appearance - within normal limits 

369-14 F S Macroscopic

external appearance - within normal limits 

369-15 F S Macroscopic

external appearance - within normal limits 

370-01 M S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

370-02 M S Macroscopic

external appearance - within normal limits 

370-03 M S Macroscopic

external appearance - within normal limits 

370-04 M S Macroscopic

external appearance - within normal limits 

370-05 M S Macroscopic

external appearance - within normal limits 

370-06 F S Macroscopic

external appearance - within normal limits 

370-07 F S Macroscopic

external appearance - within normal limits 

370-08 F S Macroscopic

external appearance - within normal limits 

370-09 F S Macroscopic

external appearance - within normal limits 

370-10 F S Macroscopic

external appearance - within normal limits 

370-11 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

150 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

370-12 F S Macroscopic

external appearance - within normal limits 

370-13 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

600 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

383-01 M S Macroscopic

external appearance - within normal limits 

383-02 M S Macroscopic

external appearance - within normal limits 

383-05 F S Macroscopic

external appearance - within normal limits 

383-06 F S Macroscopic

external appearance - within normal limits 

383-07 F S Macroscopic

external appearance - within normal limits 

383-09 F S Macroscopic

external appearance - within normal limits 

383-10 F S Macroscopic

external appearance - within normal limits 

383-11 F S Macroscopic

external appearance - within normal limits 

383-12 F S Macroscopic

external appearance - within normal limits 

384-12 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

2400 mg/kg/day

S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

389-01 M S Macroscopic

external appearance - within normal limits 

389-02 M S Macroscopic

external appearance - within normal limits 

389-03 M S Macroscopic

external appearance - within normal limits 

389-04 F S Macroscopic

external appearance - within normal limits 

389-05 F S Macroscopic

external appearance - within normal limits 

389-06 F S Macroscopic

external appearance - within normal limits 

389-07 F S Macroscopic

external appearance - within normal limits 

389-08 F S Macroscopic

external appearance - within normal limits 

389-09 F S Macroscopic

external appearance - within normal limits 

389-10 F S Macroscopic

external appearance - within normal limits 
MPI Research Study Number 1271-001 

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

Individual F1 Pup Animal Data Record: Pathology 

Day 4 Pups 

Group,

Animal Number Sex Fate Tissue Observations

2400 mg/kg/day

 S - Scheduled Necropsy E - Euthanized in extremis D - Died on Study U – Unable to Determine Sex NT – Not Tattooed

389-11 F S Macroscopic

external appearance - within normal limits 

389-12 F S Macroscopic

external appearance - within normal limits 
1271-001 

Appendix R 

Protocol and Amendments 

Page 332 of 358
 

AN ORAL REPRODUCTION/DEVELOPMENTAL TOXICITY 

SCREENING STUDY OF ANTHRAQUINONE (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 F1 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 (Levene’s/ANOVA-Dunnett’s/Welch’s) ........16 

9.2. Arcsin-Square-Root Transformation 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 Screening 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 test article on male and female 

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

conception, development of the conceptus, and parturition in rats. 

1.6. Regulatory Guideline 

This protocol meets or exceeds Guideline 421, Reproduction/Developmental Toxicity Screening 

Test, the Organization of Economic Cooperation and Development (OECD) Guideline for 

Testing of Chemicals, revised July 1995. 

1.7. Good Laboratory Practice 

This nonclinical laboratory study will be conducted in accordance with the United States 

Environmental Protection Agency (EPA), Toxic Substance Control Act (TSCA) Good 

Laboratory Practice (GLP) Standards, 40 CFR 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 Association for Assessment and Accreditation of 

Laboratory Animal Care International (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 

ProvantisTM: 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. 

Millennium32: Millennium32 HPLC System (instrument) and 

Millennium32 Chromatography Manager Version 

(software) used to quantitatively determine the amount 

of test article in prepared test article batches. 

MPI Archiving System (MArcS): In-house developed 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 ProvantisTM data. 

Master Schedule: Maintains the master 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 applications development. 

Microsoft® Office 2003 Professional: Bundle of integrated productivity tools including word 

and data processing and communications 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 Study 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 inspections and the draft and final reports will be 

reviewed by the Quality Assurance Department of MPI Research in accordance with MPI 

Research’s Standard Operating Procedures. Study quality assurance inspection records will be 

made available to the Sponsor Representatives during visits to MPI Research. 

1.13. Alteration of Design 

Alterations of this protocol may be made as the study progresses. No changes in the protocol 

will be made without the specific written 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 Director 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 and 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, storage conditions, expiration date, safe handling procedures, physical 

properties, as well as other relevant information 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 article, 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 diluted 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 the test article will be 

mixed with an appropriate vehicle to achieve the desired concentrations. The vehicle and 

method of preparation will be determined based upon physical characteristics of the test article 

and size of batches required. Fresh suspensions will be prepared for each concentration weekly, 

unless the stability analysis results indicate otherwise. 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, test batches of the test article 

formulation at both the low and high concentrations will be prepared and analyzed. The test 

batches will be prepared employing the same method 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 suspensions 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 results of this analysis are 

unsatisfactory, further testing may be necessary to obtain satisfactory stability 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 analysis of test article concentration. Samples will be stored 

refrigerated until analyzed. Following completion 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 MPI 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 multiple studies are conducted with the same test article, a common reserve 

sample may be taken and 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 article 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, storage conditions, expiration date, safe handling procedures, physical 

properties, as well as other relevant information 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 information 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 applicable 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 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 interactions between the various cells, 

tissues, and organs comprising the human body".2

 

The rat is the usual rodent model3

 used for evaluating the toxicity of various classes of chemicals 

and for which there is a large 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 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 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 

F1 pups (based on approximately 14 pups per litter): 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 the fewest number of animals possible, consistent with the 

objective of the study, the scientific needs of the Sponsor, contemporary scientific 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 within this weight range to satisfy the number of 

animals required to go on study, the Study Director will be notified to ascertain the appropriate 

action to be taken. 

Animals considered suitable for study will be weighed 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 randomized, by sex, into treatment groups using a 

standard, by weight, block randomization procedure. 

3.7.5. Method of Identification 

Each animal will be assigned an animal number to be used in ProvantisTM and will be implanted 

with a microchip bearing a unique identification number. Pups will be identified by tattoo on 

approximately Lactation Day (LD) 0. The individual 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 number, 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 clinical 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 selection for study. All animals with any evidence 

of disease or physical abnormalities will not be selected 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 suspended, stainless steel, wire-mesh type cages, 

except during pairing, near parturition, and during lactation. During pairing, animals will be 

housed 2/cage. On approximately Gestation Day (GD) 20, females will be individually housed 

in plastic cages containing wood chip bedding until euthanasia on LD 4. 

3.8.3. Environmental Conditions 

Fluorescent lighting will be provided 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 recorded 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, PMI 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 likely to be present in the certified 

diet that would interfere with the results of this study. Therefore, no analyses other than those 

routinely performed by the feed 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 Procedures. 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 than 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 will be administered by gavage. 

5.2. Justification for Route of Administration 

The oral route is one of the potential routes of human exposure 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 mating and continue to terminal euthanasia (14 

day mating period and 14 days post-mating). Dosing of the females will begin 14 days prior to 

mating and continue through LD 4. Unmated females 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 administered via gavage. The test and control article 

preparations will be stirred continuously throughout dosing. The control animals will receive the 

control article at the same volume as the test article. 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 least twice a day, 7 days a week, for morbidity, mortality, and signs 

of injury. Any findings will be recorded. Should 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, abdomen, 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. During the mating period, food consumption will not be measured 

because the animals will be cohabitating at this time. Food consumption 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 administration 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 females during the mating period and the presence or 

absence of sperm or vaginal plug will be recorded. 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, any female with no positive evidence of mating 

will be individually housed in a plastic cage with 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 F1 Litter Observations 

Toward the end of the gestation period, females will be examined twice daily for signs of 

parturition. The mated females will be allowed to give birth (F1). The duration of gestation will 

Page 344 of 358
MPI Research Study Number: 1271-001

13 

be calculated, and any difficulties occurring at parturition 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 following 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 euthanized and evaluated as described in the 

Anatomic Pathology section. 

7. EUTHANASIA 

7.1. Moribundity 

Any moribund animals, as defined by the Testing Facility 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 inhalation followed by an MPI Research SOP approved 

method to ensure death, e.g. exsanguination. Euthanasia of fetuses/pups (LD 0-4) will be by an 

intraperitoneal injection of sodium pentobarbital 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 animal will be documented in the study records. 

8. POSTMORTEM EVALUATIONS 

8.1. Anatomic Pathology 

Complete necropsy examinations will be performed 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 dead will be necropsied as soon as possible. If 

an animal cannot be necropsied immediately after being found (within 2 hours), the carcass will 

be refrigerated until the necropsy is performed. 

At the termination of the study, all surviving adult animals will be euthanized and examined. The 

animals will be examined carefully 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 findings. The abdominal, thoracic, and cranial 

cavities will be examined for abnormalities. Special 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 counted 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 testis will be weighed separately, and a combined 

weight will be presented in the report. For processing of the Bouins-fixed testes, hematoxylin 

and periodic acid Schiff (PAS) staining and paraffin embedding will be performed. Plastic 

(methacrylate) embedding of the testes and epididymides and staging of testes for 

spermatogenesis can be conducted at the Sponsor’s request (extra cost). 

Body weight and the organ weights identified in the following table will be recorded for all 

animals at scheduled necropsies and appropriate 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 specified otherwise. Microscopic examination of 

formalin-fixed hematoxylin and eosin-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 Xc

 gross lesions X X 

 identified target organsb X X Xc

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 fetuses (if present) from the animals listed above will 

be conducted by Developmental Toxicology personnel under the supervision of a veterinary 

pathologist, and the intact fetuses will be saved in 10% neutral buffered formalin for possible 

future examination. 

F1 pups from dams dying during lactation 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 externally examined, and any abnormalities will be 

recorded. The carcasses will then be discarded. 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 other 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 statistical analyses described 

below. If more than one set of comparisons is required, all analyses will be conducted separately 

on each set unless stated otherwise. Data for each sex within a set 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 Changes Group Pair-wise Comparisons 

 Gestation Food Consumption Group Pair-wise Comparisons 

 Lactation Body Weights Group Pair-wise Comparisons 

 Lactation Body Weight Changes Group Pair-wise Comparisons 

 Lactation Food Consumption Group Pair-wise Comparisons 

 Postmating Body Weights-Males Group Pair-wise Comparisons 

 Postmating Food Consumption-Males Group Pair-wise Comparisons 

Fertility Indices 

 Estrous Cycle (mean cycle time) Group Pair-wise Comparisons 

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

 Gestation Length Group 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 Weights Group Pair-wise Comparisons 

 Female Reproduction Organ Weights Group Pair-wise Comparisons 

 

Litter 

 Litter Size Group Pair-wise Comparisons 

 Viable Pups Group Pair-wise Comparisons 

 Pup Sex Ratio (% viable males/litter) 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 test4 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 analysis of variance (ANOVA) and utilized by a 

Dunnett’s5 comparison of each treatment group with the control group. If Levene’s test is 

significant (p<0.01), comparisons with the control group will be made using Welch’s t-test6 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-test7. 

Results of all pair-wise comparisons will be reported at the 0.05 and 0.01 significance levels. All 

endpoints will be analyzed using two-tailed tests unless indicated otherwise. 

9.2. Arcsin-Square-Root Transformation Followed by Group-Pairwise Comparisons 

Data comprised of percent values will be transformed using the arcsin of the square root7

. The 

analysis described in the Group Pair-Wise Comparison paragraph will 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 comparing several treatments with a control. J Am Stat Assoc 

1955;50:1096-1121. 

6

 Welch BL. The significance of difference between two means when the population variances are unequal. 

Biometrika 1937;29:350-362. 

7

 Steel RGD, Torrie JH. Principles and Procedures of Statistics. A biometrical 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 

exact8 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 treatment group will be compared to the control 

group using a Fisher’s exact test. Results will be reported at the 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-wise testing will be conducted for 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 covariance9

 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 will be performed. Each treatment satisfying the 

sample size assumption will be compared to control using Dunnett’s test under 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 reported at the 0.05 and 0.01 significance levels. 

Endpoints will be analyzed using two-tailed 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 draft report containing the results of all tests, 

analyses, observations and measurements required by this protocol, and an interpretative 

summary of the study results will be submitted to the Sponsor. The report will include all items 

required by the applicable regulatory agency. After 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 revisions or instructions to finalize 

have been communicated by the Sponsor, the draft report 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 draft will be performed at additional cost to the 

Sponsor. 

 

8

 Zar JH. Biostatistical Analysis. 4th ed. New Jersey: Prentice Hall; 1999. 9

 Snedecor, GW, Cochran, WG. Statistical Methods, Eighth 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, reserve samples, specimens (slides, blocks, and 

wet tissues), and the final report generated as a result of this study will 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 (final report issue date). Retention of materials after the time 

stated above will be subject to a future contractual agreement between the Sponsor and MPI 

Research. 

12. ANIMAL WELFARE 

MPI Research is committed to complying with all applicable regulations 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 Welfare Act (9 CFR Parts 1, 2 and 3). The Guide 

for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National 

Academy Press, Washington, D.C., 1996, will be followed. This facility maintains an Animal 

Welfare Assurance statement with National Institutes of Health, Office of Laboratory Animal 

Welfare. 

To ensure compliance: 

A. This protocol will be reviewed and approved by the Institutional Animal Care and Use 

Committee (IACUC) before animal receipt or transfer. 

B. The Sponsor, by his or her signature, attests that the activities specified in this protocol do not 

unnecessarily duplicate any previous experiment. 

C. The Study Director has considered alternatives 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 another 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 testing methods; alternative testing methods, 

toxicology; general toxicology testing method alternatives 

Databases searched: toxnet.nlm.nih.gov;pubmed.gov;medscape.com;caat.jhsph.edu 

 Date: February 14, 2006 

Interval Searched: All years to Present 

Search terms: alternative testing methods rodents; alternative toxicology methods 

rodents; rodents, alternative testing; 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 two 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. cageside observation was not documented. 

On July 4, 2006, test article administration was conducted 11 minutes 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 formalin for possible further examination. 

In the opinion of the Study Director, these minor deviations did not affect the quality or 

integrity of the study. 

Page 358 of 358