NTP TECHNICAL REPORT
 

ON THE
 

TOXICOLOGY AND CARCINOGENESIS
 

STUDIES OF
 

D&C YELLOW NO. 11
 

(CAS NO. 8003-22-3) 

IN F344/N RATS 

(FEED STUDIES) 

NATIONAL TOXICOLOGY PROGRAM
 

P.O. Box 12233
 

Research Triangle Park, NC 27709
 

April 1997 

NTP TR 463
 

NIH Publication No. 97-3379
 

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES
 

Public Health Service
 

National Institutes of Health
 
FOREWORD 

The National Toxicology Program (NTP) is made up of four charter agencies of the U.S. Department of Health and 

Human Services (DHHS): the National Cancer Institute (NCI), National Institutes of Health; the National 

Institute of Environmental Health Sciences (NIEHS), National Institutes of Health; the National Center for 

Toxicological Research (NCTR), Food and Drug Administration; and the National Institute for Occupational 

Safety and Health (NIOSH), Centers for Disease Control. In July 1981, the Carcinogenesis Bioassay Testing 

Program, NCI, was transferred to the NIEHS. The NTP coordinates the relevant programs, staff, and resources 

from these Public Health Service agencies relating to basic and applied research and to biological assay 

development and validation. 

The NTP develops, evaluates, and disseminates scientific information about potentially toxic and hazardous 

chemicals. This knowledge is used for protecting the health of the American people and for the primary prevention 

of disease. 

The studies described in this Technical Report were performed under the direction of the NIEHS and were 

conducted in compliance with NTP laboratory health and safety requirements and must meet or exceed all 

applicable federal, state, and local health and safety regulations. Animal care and use were in accordance with the 

Public Health Service Policy on Humane Care and Use of Animals. The prechronic and chronic studies were 

conducted in compliance with Food and Drug Administration (FDA) Good Laboratory Practice Regulations, and 

all aspects of the chronic studies were subjected to retrospective quality assurance audits before being presented 

for public review. 

These studies are designed and conducted to characterize and evaluate the toxicologic potential, including 

carcinogenic activity, of selected chemicals in laboratory animals (usually two species, rats and mice). Chemicals 

selected for NTP toxicology and carcinogenesis studies are chosen primarily on the bases of human exposure, level 

of production, and chemical structure. The interpretive conclusions presented in this Technical Report are based 

only on the results of these NTP studies. Extrapolation of these results to other species and quantitative risk 

analyses for humans require wider analyses beyond the purview of these studies. Selection per se is not an 

indicator of a chemical’s carcinogenic potential. 

These NTP Technical Reports are available for sale from the National Technical Information Service, 

U.S. Department of Commerce, 5285 Port Royal Road, Springfield, VA 22161 (703-487-4650). Single copies of 

this Technical Report are available without charge while supplies last from NTP Central Data Management, 

NIEHS, P.O. Box 12233, MD E1-02, Research Triangle Park, NC 27709 (919-541-3419). Listings of all 

published NTP reports and ongoing studies are also available from NTP Central Data Management. The 

Abstracts and other study information for 2-year studies are also available at the NTP’s World Wide Web site: 

http://ntp-server.niehs.nih.gov. 
NTP TECHNICAL REPORT
 

ON THE
 

TOXICOLOGY AND CARCINOGENESIS
 

STUDIES OF
 

D&C YELLOW NO. 11
 

(CAS NO. 8003-22-3) 

IN F344/N RATS 

(FEED STUDIES) 

NATIONAL TOXICOLOGY PROGRAM
 

P.O. Box 12233
 

Research Triangle Park, NC 27709
 

April 1997 

NTP TR 463
 

NIH Publication No. 97-3379
 

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES
 

Public Health Service
 

National Institutes of Health
 
2 D&C Yellow No. 11, NTP TR 463 

CONTRIBUTORS
 

National Toxicology Program 

Evaluated and interpreted results and reported findings 

W.C. Eastin, Ph.D., Study Scientist 

G.A. Boorman, D.V.M., Ph.D. 

D.A. Bridge, B.S. 

J.R. Bucher, Ph.D. 

L.T. Burka, Ph.D. 

R.E. Chapin, Ph.D. 

J.R. Hailey, D.V.M. 

J.K. Haseman, Ph.D. 

A. Radovsky, D.V.M., Ph.D. 

G.N. Rao, D.V.M., Ph.D. 

J.H. Roycroft, Ph.D. 

G.S. Travlos, D.V.M. 

D.B. Walters, Ph.D. 

K.L. Witt, M.S., Oak Ridge Associated Universities 

Southern Research Institute 

Conducted studies, evaluated pathology findings 

J.D. Prejean, Ph.D., Principal Investigator 

D.G. Serota, Ph.D., Principal Investigator 

J.E. Heath, D.V.M. 

R. Hoar, Ph.D., Argus Research Laboratories, Inc. 

C. Lindamood, III, Ph.D. 

Experimental Pathology Laboratories, Inc. 

Provided pathology quality assurance 

J.F. Hardisty, D.V.M., Principal Investigator 

C.C. Shackelford, D.V.M., M.S., Ph.D. 

Dynamac Corporation 

Prepared quality assurance audits 

S. Brecher, Ph.D., Principal Investigator 

Analytical Sciences, Inc. 

Provided statistical analyses 

R.W. Morris, M.S., Principal Investigator 

N.G. Mintz, B.S. 

S. Rosenblum, M.S. 

NTP Pathology Working Group 

Evaluated slides, prepared pathology report on rats (4 April 1995) 

M.P. Jokinen, D.V.M., Chairperson 

Pathology Associates, Inc. 

R.C. Cattley, M.S., V.M.D., Ph.D. 

Chemical Industry Institute of Toxicology 

M.R. Elwell, D.V.M., Ph.D. 

National Toxicology Program 

L. Gaboury, M.D., Ph.D., Observer 

Universite de Montreal 

J.R. Hailey, D.V.M. 

National Toxicology Program 

R.A. Herbert, D.V.M., Ph.D. 

National Toxicology Program 

R.R. Maronpot, D.V.M. 

National Toxicology Program 

A. Radovsky, D.V.M., Ph.D. 

National Toxicology Program 

C.C. Shackelford, D.V.M., M.S., Ph.D. 

Experimental Pathology Laboratories, Inc. 

Evaluated slides, prepared pathology report on extended renal 

evaluation in rats (19 September 1995) 

J.F. Hardisty, D.V.M., Chairperson 

Experimental Pathology Laboratories, Inc. 

D. Dixon, D.V.M., Ph.D. 

National Toxicology Program 

J. Hellman, D.V.M., Observer 

National Toxicology Program 

D.E. Malarkey, D.V.M. 

National Toxicology Program 

R.R. Maronpot, D.V.M. 

National Toxicology Program 

A. Radovsky, D.V.M., Ph.D. 

National Toxicology Program 

R.C. Sills, D.V.M., Ph.D. 

National Toxicology Program 

Biotechnical Services, Inc. 

Prepared Technical Report 

S.R. Gunnels, M.A., Principal Investigator 

L.M. Harper, B.S. 

D.C. Serbus, Ph.D. 

S.M. Swift, B.S. 
3 

CONTENTS
 

ABSTRACT ................................................................................. 5
 

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY ................... 9
 

TECHNICAL REPORTS REVIEW SUBCOMMITTEE .......................................... 10
 

SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS ............... 11
 

INTRODUCTION ............................................................................ 13
 

MATERIALS AND METHODS ............................................................... 17
 

RESULTS ................................................................................... 27
 

DISCUSSION AND CONCLUSIONS ........................................................... 45
 

REFERENCES .............................................................................. 49
 

APPENDIX A Summary of Lesions in Male Rats in the 2-Year Feed Study
 

of D&C Yellow No. 11 .......................................................... 53
 

APPENDIX B Summary of Lesions in Female Rats in the 2-Year Feed Study
 

of D&C Yellow No. 11 .......................................................... 99
 

APPENDIX C Genetic Toxicology ............................................................ 139
 

APPENDIX D Organ Weights and Organ-Weight-to-Body-Weight Ratios ......................... 153
 

APPENDIX E Hematology Results ............................................................ 155
 

APPENDIX F Reproductive Toxicity Study Results ............................................. 157
 

APPENDIX G Chemical Characterization and Dose Formulation Studies .......................... 163
 

APPENDIX H Feed and Compound Consumption in the 2-Year Feed Study
 

of D&C Yellow No. 11 .......................................................... 175
 

APPENDIX I Ingredients, Nutrient Composition, and Contaminant Levels 

in NIH-07 Rat and Mouse Ration ................................................ 179
 

APPENDIX J Sentinel Animal Program ...................................................... 183
 
4 D&C Yellow No. 11, NTP TR 463 
O 

C N C 

C 

H 

O 

5 

ABSTRACT
 

D&C YELLOW NO. 11 

CAS No. 8003-22-3
 

Chemical Formula: C 18 11 2 H NO Molecular Weight: 273.29

Synonyms:	 2-(2-Quinolinyl)-1H-indene-1,3-(2H)-dione; 2-(2-quinolyl)-1,3-indandione 

Trade names:	 Arlosol Yellow S, Chinoline Yellow D (soluble in spirits), Chinoline Yellow ZSS, C.I. 47000, C.I. Solvent Yellow 33, Nitro Fast 

Yellow SL, Oil Yellow SIS, Petrol Yellow C, Quinoline Yellow A Spirit Soluble, Quinoline Yellow Base, Quinoline Yellow Spirit 

Soluble, Quinoline Yellow SS, Solvent Yellow 33, Waxoline Yellow T 

D&C Yellow No. 11 is used to color topical drug 

preparations and cosmetics. It is also used in spiri t 

lacquers, polystyrenes, polycarbonates, polyamides , 

acrylic resins, colored smokes, and hydrocarbon sol ­

vents. D&C Yellow No. 11 was nomin ated to the NTP 

for toxicity and carcinogenesis studies as part of a 

larger regulatory effort mandated by Congress an d 

undertaken by the Food and Drug Administration t o 

determine the safety of a number of provisionally listed 

dyes. D&C Yellow No. 11 is currently regulated fo r 

external use. The recommendation to study D& C 

Yellow No. 11 by dietary exposure was based on th e 

fact that it is a contaminant of D&C Yellow No. 10, a 

candidate for permanent listing as a chemical for which 

there is a potential for ingestion. 

First-generation (F ) male and female F344/N rat s 0 

were given D&C Yellow No. 11 (approximately 99% 

pure) in feed for up to 19 weeks and then mated, an d 

exposure of second-generation (F ) males and females 1 

began in utero and continued for 2 years after weaning 

at 28 days of age. Genetic toxicology studies wer e 

conducted in Salmonella typhimurium , cultured 

Chinese hamster ovary cells, and mouse periphera l 

blood. 

REPRODUCTIVE TOXICITY STUDY 

Groups of 60 male and 60 female F rats were given 0, 0 

500, 1,700, or 5,000 ppm D&C Yellow No. 11 in feed 

for up to 19 weeks, which resulted in average dail y 

doses of 35, 120, or 350 mg D&C Yellow No. 11/kg 

body weight to males and 35, 120, or 370 mg/kg t o 

females. All F0 males and females survived until th e

end of the study. Prior to cohabitation, mean bod y 

weight gains of males given 500, 1,700, or 5,000 ppm 

and of females given 5,000 ppm were significantl y 

lower than those of the controls. The mean bod y 

weight gains of exposed females during gestation and 

lactation were generally similar to those of the con ­

trols. Feed consumption by exposed groups of rat s 

was generally similar to that by the control group s 

prior to cohabitation. 

The duration of gestation, the average litter size, th e 

number of live pups on days 4 (precull) and 21, and 
6 D&C Yellow No. 11, NTP TR 463 

the percentage of male pups for each exposure group 

were similar to those of the controls. The mean body 

weights of exposed litters were significantly less than 

those of the control litters on days 14 and 21; thi s 

effect was considered to be related to D&C Yellow No. 

11 exposure. 

2-YEAR STUDY 

Groups of 60 male and 60 female F rats were given 0, 1 

500, 1,700, or 5,000 ppm D&C Yellow No. 11 in feed 

for 105 (males) or 106 (females) weeks after weaning 

(day 28); 6 to 10 rats per group were evaluated at 12 

months. These exposure concentrations resulted i n 

average daily doses of approximately 25, 85, o r 

250 mg D&C Yellow No. 11/kg body weight to males 

and 25, 100, or 280 mg/kg to females. 

Survival, Body Weights, Feed Consumption, 

and Clinical Findings 

Survival of males given 1,700 or 5,000 ppm was significantly less than that of the controls, and survival of 

1,700 ppm females was significantly greater than that 

of the controls. Mean body weights of 1,700 an d 

5,000 ppm males and females were generally lowe r 

than those of the controls throughout the study. Feed 

consumption by exposed groups was similar to that by 

the controls. Chemical-related clinical findings in ­

cluded yellow discoloration of the entire body in al l 

exposed males and females from day 1 and hea d 

swelling and edema in 1,700 and 5,000 ppm males . 

One 1,700 ppm and five 5,000 ppm males were moribund and were killed between weeks 49 and 81; these 

deaths were attributed to extensive edema. 

Hematology 

A few minimal hematology changes occurred in male 

rats at the 12-month interim evaluation. There wa s 

evidence of minimal anemia in exposed males; thi s 

anemia was characterized by decreased hematocri t 

values, hemoglobin concentrations, and erythrocyt e 

counts. The minimal anemia was characterized a s 

normocytic, normochromic, and nonresponsive. There 

were no biologically or statistically significant differences in hematology parameters between control an d 

exposed females. 

Pathology Findings 

Absolute and relative liver weights of all expose d 

groups of males and females were significantly greater 

than those of the controls at 12 months. At 2 years , 

the incidences of hepatocellular adenoma in 5,000 ppm 

males and of hepatocellular adenoma or carcinom a 

(combined) in 5,000 ppm females were significantl y 

greater than those in the controls. At 12 months, th e 

incidences of clear cell foci in 1,700 and 5,000 pp m 

females were significantly greater than that in th e 

controls. At 2 years, the incidences of mixed cell foci 

in exposed males and of clear cell foci in expose d 

males (except 500 ppm) and females were significantly 

greater than those in the controls. Incidences o f 

cytologic alterations (basophilia and granularity) o f 

hepatocytes, and pigmentation in bile duct epithelium, 

hepatocytes, and Kupffer cells in exposed males an d 

females were greater than those in the controls at both 

12 months and 2 years. 

Renal tubule adenomas were observed in tw o 

5,000 ppm males, and one renal tubule carcinoma was 

observed in a 1,700 ppm male. During an extende d 

evaluation, renal tubule adenomas were observed i n 

two additional 5,000 ppm males, four 1,700 pp m 

males, and two 500 ppm males. Renal tubule hyper ­

plasia was observed in exposed groups of males bu t 

not in controls, and the incidences in 1,700 ppm males 

from both standard and extended evaluations wer e 

significantly greater than those in the controls. Necrosis and regeneration of the renal tubule epitheliu m 

were observed in all control and exposed male rats and 

in most female rats at 12 months and 2 years. Th e 

severity of nephropathy in exposed males and females 

was significantly greater than that in the controls. I n 

exposed males and 1,700 ppm females at 2 years, the 

incidences of hyperplasia of the transitional epithelium 

in the kidney, which commonly accompanies advanced 

nephropathy, were greater than those of the controls , 

and the severity of this lesion in exposed males an d 

females was greater than that in the controls. Th e 

incidences of renal tubule pigmentation in all exposed 

groups of males and females at 12 months and 2 years 

were significantly greater than those in the controls. 

Squamous cell carcinomas of the tong ue were observed 

in one 500 ppm male at 12 months and one 5,000 ppm 

female at 2 years, and one squamous cell carcinoma of 

the oral mucosa was observed in each group of ex ­

posed males and in one 5,000 ppm female at 2 years. 

At 2 years, squamous cell papi llomas were observed in 

the oral cavity (oral mucosa or tongue) of one control, 

one 500 ppm, two 1,700 ppm, and four 5,000 pp m 
__________ 

D&C Yellow No. 11, NTP TR 463 7 

males; this lesion was also observed in one control and some evidence of carcinogenic activity * of D&C 

one 500 ppm female. Yellow No. 11 in male F344/N rats based on increased 

incidences of hepatocellular adenoma, renal tubul e 

GENETIC TOXICOLOGY 

Results of mutagenicity tests with D &C Yellow No. 11 

in Salmonella typhimurium were equivocal in one 

study, based on responses observed in strain TA10 0 

with induced rat liver S9, and weakly positive in a 

second study, based on responses observed in strain s 

TA98 and TA100 with induced rat or hamster liver S9. 

D&C Yellow No. 11 induced sister chromatid ex ­

changes and chromosomal aberrations in culture d 

Chinese hamster ovary cells, with and without S9. No 

increase in the frequency of micronucleate d 

normochromatic erythrocytes was observed i n 

peripheral blood samples from male and femal e 

B6C3F1 mice administered D&C Yellow No. 11 i n 

feed for 13 weeks. 

CONCLUSIONS 

Under the conditions of this perinatal exposur e 

followed by a 2-year dosed feed study, there was 

neoplasms, and squamous cell neoplasms of the ora l 

cavity. There was some evidence of carcinogeni c 

activity in female F344/N rats based on increased incidences of hepatocellular neoplasms. Incidences o f 

uncommon squamous cell carcinoma of the oral cavity 

in females may have been related to chemica l 

treatment. 

Exposure of rats to D&C Yellow No. 11 in feed for 2 

years resulted in increased incidences of nonneoplastic 

liver lesions including clear cell foci, increase d 

basophilia and granularity in the cytoplasm of hepatocytes, and bile duct, hepatocyte, and Kupffer cel l 

pigmentation in males and females and mixed cell foci 

in males. In the kidney, there were increase d 

incidences of renal tubule pigmentation and transi ­

tional epithelial hyperplasia in males and females and 

renal tubule hyperplasia in males. The severity o f 

nephropathy was increased in exposed males an d 

females. 

* Explanation of Levels of Evidence of Carcinogenic Activity is on page 9. A summary of the Technical Reports Review Subcommittee comments and 

the public discussion on this Technical Report appears on page 11. 
8 D&C Yellow No. 11, NTP TR 463 

Summary of the 2-Year Carcinogenesis and Genetic Toxicology Studies of D&C Yellow No. 11 

Male F344/N Rats	 Female F344/N Rats 

Doses	 0, 500, 1,700, or 5,000 ppm 0, 500, 1,700, or 5,000 ppm 

Body weights	 1,700 and 5,000 ppm groups lower than control 1,700 and 5,000 ppm groups lower than control 

group group 

2-Year survival rates	 19/50, 20/51, 8/51, 2/54 22/50, 26/51, 37/50, 23/51 

Nonneoplastic effects	 Liver: clear cell focus (9/50, 15/51, 15/51, 18/54); Liver: clear cell focus (10/50, 18/51, 29/50, 

mixed cell focus (1/50, 10/51, 9/51, 10/54); bile 30/51); bile duct pigmentation (0/50, 46/51, 

duct pigmentation (0/50, 38/51, 51/51, 54/54); 49/50, 50/51); hepatocyte cytologic alterations 

hepatocyte cytologic alterations (0/50, 20/51, 44/51, (0/50, 11/51, 31/50, 40/51); hepatocyte 

42/54); hepatocyte pigmentation (0/50, 22/51, pigmentation (0/50, 34/51, 44/50, 50/51); Kupffer 

45/51, 51/54); Kupffer cell pigmentation (7/50, cell pigmentation (9/50, 11/51, 16/50, 32/51) 

15/51, 23/51, 26/54) Kidney: renal tubule pigmentation (10/50, 48/51, 

Kidney: renal tubule hyperplasia (standard 50/50, 51/51); transitional epithelial hyperplasia 

evaluation – 0/50, 0/51, 4/51, 3/54; extended (2/50, 6/51, 10/50, 3/51); severity of nephropathy 

evaluation – 0/50, 2/51, 9/51, 2/54; standard and (1.4, 1.7, 1.8, 2.1) 

extended evaluations combined – 0/50, 2/51, 13/51, 

4/54); renal tubule pigmentation (18/50, 43/51, 

47/51, 54/54); transitional epithelial hyperplasia 

(11/50, 23/51, 29/51, 34/54); severity of 

nephropathy (2.3, 2.8, 3.2, 3.0) 

Neoplastic effects	 Liver: hepatocellular adenoma (1/50, 2/51, 1/51, Liver: hepatocellular adenoma or carcinoma 

7/54) (0/50, 2/51, 5/50, 5/51) 

Kidney: renal tubule adenoma (standard 

evaluation – 0/50, 0/51, 0/51, 2/54; extended 

evaluation – 0/50, 2/51, 4/51, 2/54; standard and 

extended evaluations combined – 0/50, 2/51, 4/51, 

4/54); renal tubule adenoma or carcinoma 

(standard and extended evaluations combined – 

0/50, 2/51, 5/51, 4/54) 

Oral cavity: squamous cell papilloma (1/50, 1/51, 

2/51, 4/54); squamous cell carcinoma (0/50, 1/51, 

1/51, 1/54); squamous cell papilloma or squamous 

cell carcinoma (1/50, 2/51, 3/51, 5/54) 

Uncertain finding None	 Oral cavity: squamous cell carcinoma (0/50, 

0/51, 0/50, 2/51); squamous cell papilloma or 

squamous cell carcinoma (1/50, 1/51, 0/50, 2/51) 

Level of evidence of Some evidence Some evidence 

carcinogenic activity 

Genetic toxicology 

Salmonella typhimurium gene mutations:	 Equivocal in strain TA100 with S9 at SRI International, and weakly 

positive in strains TA98 and TA100 with S9 at Microbiological 

Associates, Inc. 

Sister chromatid exchanges
 

Cultured Chinese hamster ovary cellsin vitro: Positive with and without S9
 

Chromosomal aberrations
 

Cultured Chinese hamster ovary cellsin vitro: Positive with and without S9
 

Micronucleated erythrocytes
 

Mouse peripheral bloodin vivo: Negative
 
D&C Yellow No. 11, NTP TR 463 9 

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY 

The National Toxicology Program describes the results of individual experiments on a chemical agent and notes the strength of the evidence for 

conclusions regarding each study. Negative results, in which the study animals do not have a greater incidence of neoplasia than control animals, do 

not necessarily mean that a chemical is not a carcinogen, inasmuch as the experiments are conducted under a limited set of conditions. Positive results 

demonstrate that a chemical is carcinogenic for laboratory animals under the conditions of the study and indicate that exposure to the chemical has the 

potential for hazard to humans. Other organizations, such as the International Agency for Research on Cancer, assign a strength of evidence for 

conclusions based on an examination of all available evidence, including animal studies such as those conducted by the NTP, epidemiologic studies, 

and estimates of exposure. Thus, the actual determination of risk to humans from chemicals found to be carcinogenic in laboratory animals requires a 

wider analysis that extends beyond the purview of these studies. 

Five categories of evidence of carcinogenic activity are used in the Technical Report series to summarize the strength of the evidence observed in each 

experiment: two categories for positive results (clear evidence and some evidence); one category for uncertain findings (equivocal evidence); one 

category for no observable effects (no evidence); and one category for experiments that cannot be evaluated because of major flaws (inadequate 

study). These categories of interpretative conclusions were first adopted in June 1983 and then revised in March 1986 for use in the Technical Report 

series to incorporate more specifically the concept of actual weight of evidence of carcinogenic activity. For each separate experiment (male rats, 

female rats, male mice, female mice), one of the following five categories is selected to describe the findings. These categories refer to the strength of 

the experimental evidence and not to potency or mechanism. 

•	 Clear evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing a dose-related (i) increase of malignant 

neoplasms, (ii) increase of a combination of malignant and benign neoplasms, or (iii) marked increase of benign neoplasms if there is an 

indication from this or other studies of the ability of such tumors to progress to malignancy. 

•	 Some evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing a chemical-related increased incidence 

of neoplasms (malignant, benign, or combined) in which the strength of the response is less than that required for clear evidence. 

•	 Equivocal evidenceof carcinogenic activity is demonstrated by studies that are interpreted as showing a marginal increase of neoplasms 

that may be chemical related. 

•	 No evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing no chemical-related increases in
 

malignant or benign neoplasms.
 

•	 Inadequate studyof carcinogenic activity is demonstrated by studies that, because of major qualitative or quantitative limitations, 

cannot be interpreted as valid for showing either the presence or absence of carcinogenic activity. 

When a conclusion statement for a particular experiment is selected, consideration must be given to key factors that would extend the actual boundary 

of an individual category of evidence. Such consideration should allow for incorporation of scientific experience and current understanding of longterm carcinogenesis studies in laboratory animals, especially for those evaluations that may be on the borderline between two adjacent levels. These 

considerations should include: 

• adequacy of the experimental design and conduct;
 

• occurrence of common versus uncommon neoplasia;
 

•	 progression (or lack thereof) from benign to malignant neoplasia as well as from preneoplastic to neoplastic lesions; 

•	 some benign neoplasms have the capacity to regress but others (of the same morphologic type) progress. At present, it is impossible to 

identify the difference. Therefore, where progression is known to be a possibility, the most prudent course is to assume that benign 

neoplasms of those types have the potential to become malignant; 

•	 combining benign and malignant tumor incidence known or thought to represent stages of progression in the same organ or tissue; 

•	 latency in tumor induction; 

•	 multiplicity in site-specific neoplasia; 

•	 metastases; 

•	 supporting information from proliferative lesions (hyperplasia) in the same site of neoplasia or in other experiments (same lesion in 

another sex or species); 

•	 presence or absence of dose relationships; 

•	 statistical significance of the observed tumor increase; 

•	 concurrent control tumor incidence as well as the historical control rate and variability for a specific neoplasm; 

•	 survival-adjusted analyses and false positive or false negative concerns; 

•	 structure-activity correlations; and 

•	 in some cases, genetic toxicology. 
 

__________ 

10 D&C Yellow No. 11, NTP TR 463 

NATIONAL TOXICOLOGY PROGRAM BOARD OF SCIENTIFIC COUNSELORS
 

TECHNICAL REPORTS REVIEW SUBCOMMITTEE
 

The members of the Technical Reports Review Subcommittee who evaluated the draft NTP Technical Report on D&C Yellow No. 11 on 

5 December 1995, are listed below. Subcommittee members serve as independent scientists, not as representatives of any institution, company, or 

governmental agency. In this capacity, subcommittee members have five major responsibilities in reviewing NTP studies: 

• to ascertain that all relevant literature data have been adequately cited and interpreted, 

• to determine if the design and conditions of the NTP studies were appropriate, 

• to ensure that the Technical Report presents the experimental results and conclusions fully and clearly, 

• to judge the significance of the experimental results by scientific criteria, and 

• to assess the evaluation of the evidence of carcinogenic activity and other observed toxic responses. 

Arnold L. Brown, M.D., Chairperson Irma Russo, M.D., Principal Reviewer 

University of Wisconsin Medical School Fox Chase Cancer Center 

Madison, WI Philadelphia, PA 

Gary P. Carlson, Ph.D., Principal Reviewer Louise Ryan, Ph.D. 

School of Health Sciences Division of Biostatistics 

Purdue University Dana-Farber Cancer Institute 

West Lafayette, IN Boston, MA 

Thomas L. Goldsworthy, Ph.D. Robert E. Taylor, M.D., Ph.D. 

Department of Experimental Pathology and Toxicology Department of Pharmacology 

Chemical Industry Institute of Toxicology Howard University College of Medicine 

Research Triangle Park, NC Washington, DC 

Robert LeBoeuf, Ph.D. Frederick L. Tyson, Ph.D. 

Corporate Professional and Regulatory Services St. Mary’s Hospital and Medical Center 

Human Safety Department Cancer Research Institute 

The Procter & Gamble Company Grand Junction, CO 

Cincinnati, OH 

Jerrold M. Ward, D.V.M., Ph.D.*

Janardan K. Reddy, M.D., Principal Reviewer* National Cancer Institute 

Department of Pathology Frederick, MD 

Northwestern University Medical School 

Chicago, IL 

* Did not attend 
D&C Yellow No. 11, NTP TR 463 11 

SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS
 

On 5 December 1995, the draft Technical Report o n 

the toxicology and carcinogenesis studies of D& C 

Yellow No. 11 received public review by the National 

Toxicology Program’s Board of Scientific Counselors’ 

Technical Reports Review Subcommittee. The review 

meeting was held at the National Institute of Environmental Health Sciences, Research Triangle Park, NC . 

Dr. W.C. Eastin, NIEHS, introduced the toxicolog y 

and carcinogenesis studies of D&C Yellow No. 11 by 

discussing the uses of the chemical and rationale fo r 

study, describing the experime ntal design, reporting on 

survival and body weight effects, and commenting on 

chemical-related neoplasms and nonneoplastic lesions 

in male and female rats. Dr. Eastin reported that th e 

study of this color additive was part of a larger effort 

mandated by Congress and undertaken by the FDA to 

determine the safety of provisionally listed dyes. The 

study design was not a standard NTP protocol. I n 

discussions with the nominator, FDA, NTP decided to 

tailor the protocol to provide perinatal exposur e 

followed by dietary exposure for 2 years in order t o 

generate data similar to those used by the FDA t o 

regulate other color additives. The proposed con ­

clusions were some evidence of carcinogenic activity 

in male and female F344/N rats . 

Dr. Reddy, a principal reviewer, was unable to attend 

the meeting but had submitted his review, whic h 

Dr. L.G. Hart, NIEHS, read into the record. Dr. Reddy 

agreed with the proposed conclusions. He said th e 

abstract should give the reasons for using only rats in 

this study. 

Dr. Russo, the second principal reviewer, agreed with 

the proposed conclusions. She also thought ther e 

needed to be clarification of why concurrent studie s 

were not done in mice. Dr. Eastin said that in pre ­

chronic studies effects in mice were about the same as 

in rats, although in all endpoints measured, rats wer e 

the more sensitive species. To have done both species 

with the larger perinatal protocol would have diverted 

resources from studying another chemical. Conducting 

the study in the more sensitive species would meet the 

FDA’s needs. 

disagreed with the perinatal protocol from this an d 

other such studies and claims made about effect s 

in utero, particularly when there are no groups ex ­

posed only post-weaning for comparison. Dr. Easti n 

agreed that these groups would have been useful a s 

well as animals exposed only in utero. Further, 

Dr. Carlson said the discussion mentions positiv e 

findings, therefore, negative findings should be cited. 

Dr. Eastin reported that there are only three other NTP 

studies with prenatal or perinatal exposures . 

Dr. Carlson said he was intrigued by the description of 

head swelling and edema and asked for mor e 

information on etiology. Dr. A. Radovsky, NIEHS , 

said the possibilities of hypoproteinemia, secondary to 

kidney or liver disease or intestinal malabsorption, and 

vascular or heart lesions were investigated. All o f 

these conditions were present in some but not in al l 

animals with edema, and the severity of kidney or liver 

neoplasms was not any greate r in these animals than in 

cohorts without edema. Thus, from an anatomi c 

histopathologic perspective, there was no explanation . 

Dr. LeBoeuf noted a body weight reduction i n 

5,000 ppm males of about 15% and wondered if thi s 

was typical or acceptable for NTP studies, rather than 

the 10%, which he thought was associated with reaching a maximum tolerated dose. Dr. J.R. Bucher , 

NIEHS, responded that it depends on the stud y 

outcome. If there is a neoplasm response in a stud y 

that has a 15% decrease, that would be acceptable , 

whereas in a negative study, such a large decreas e 

might help prevent development of a neoplasti c 

response. 

Dr. Bucher reported that in 5,000 ppm female rats, a 

second unusual oral cavity carcinoma was observed . 

Thus, NTP proposed adding a sentence to the end o f 

the first paragraph of the conclusions, but the primary 

level of evidence in female rats would remain some 

evidence of carcinogenic activity . 

Dr. Russo moved that the Technical Report on D& C 

Yellow No. 11 be accepted with the revision s 

discussed and the conclusions as written for male and 

female rats. Dr. Carlson seconded the motion, which 

was accepted with six yes votes and one abstentio n 

Dr. Carlson, the third principal reviewer, agreed with (Dr. LeBoeuf). 

the proposed conclusions. He commented that h e 
12 D&C Yellow No. 11, NTP TR 463 
O 

C N C 

C 

H 

O 

13 

INTRODUCTION
 

D&C YELLOW NO. 11 

CAS No. 8003-22-3
 

Chemical Formula: C 18 11 2 H NO Molecular Weight: 273.29

Synonyms:	 2-(2-Quinolinyl)-1H-indene-1,3-(2H)-dione; 2-(2-quinolyl)-1,3-indandione 

Trade names:	 Arlosol Yellow S, Chinoline Yellow D (soluble in spirits), Chinoline Yellow ZSS, C.I. 47000, C.I. Solvent Yellow 33, Nitro Fast 

Yellow SL, Oil Yellow SIS, Petrol Yellow C, Quinoline Yellow A Spirit Soluble, Quinoline Yellow Base, Quinoline Yellow Spirit 

Soluble, Quinoline Yellow SS, Solvent Yellow 33, Waxoline Yellow T 

CHEMICAL AND PHYSICAL PROPERTIES 

D&C Yellow No. 11 is a bright greenish yellow solid or 

a canary yellow powder with a melting point range of 

240.9 to 242.1 C. It is soluble in acetone, benzene, 

chloroform, toluene, and xylene; slightly soluble in 

methanol, ethanol, ethyl acetate, linseed oil, mineral oil, 

oleic acid, paraffin wax, stearic acid, and turpentine; and 

insoluble in water (Colour Index, 1982; Merck Index, 

1989). D&C Yellow No. 11 is the name given to 2-(2­

quinolyl)-1,3-indandione when it meets United States 

Certification Regulations (21 CFR, §74.1711). These 

regulations state that the certified dye must conform to 

the following specifications and be free from impurities 

other than those named: volatile matter 1%, matter 

insoluble in ethyl alcohol 0.4%, phthalic acid 0.3%, 

quinaldine 0.2%, subsidiary colors 5%, lead 

20 ppm, arsenic 3 ppm, and mercury 1 ppm. D&C 

Yellow No. 11 does not contain the methylated congener 

6-methyl-2-(2-quinolyl)-1,3-indandione. However, the 

noncertified dye, usually referred to as Solvent Yellow 33 

(CTFA, 1982), is composed of two parts nonmethylated 

and one part methylated forms of the dye (Colour Index, 

1982). In some toxicity studies, the D&C Yellow No. 11 

that was used contained both 2-(2-quinolyl)-1,3­

indandione and 6-methyl-2-(2-quinolyl)-1,3-indandione 

(Björkner and Niklasson, 1983; Weaver, 1983; Sato et 

al., 1984). 

PRODUCTION, USE, AND 

HUMAN EXPOSURE 

D&C Yellow No. 11 is generally used in solvent form to 

color topical drug preparations and cosmetics (CTFA, 

1982; El Dareer et al., 1988). In the United States, D&C 

Yellow No. 11 is approved only for external applications 

(21 CFR, §74.1711; Marmion, 1991). It is also used in 

spirit lacquers, polystyrenes, polycarbonates, polyamides, 

acrylic resins, colored smokes, and occasionally 

hydrocarbon solvents (Merck Index, 1989). Between

1985 and 1995, 24,580 pounds of D&C Yellow No. 11 

were certified, and 131 cosmetic formulations containing 

the dye were reported (FDA, personal communication). 

The National Occupational Exposure Survey estimated 

that 14,313 workers (4,310 females) were potentially 

exposed to D&C Yellow No. 11 in five different 

industries from 1981 to 1983 (NIOSH, 1990). 
14 D&C Yellow No. 11, NTP TR 463 

ABSORPTION, DISTRIBUTION, 

METABOLISM, AND EXCRETION 

Experimental Animals 

Studies sponsored by the NTP demonstrated tha t 

radiolabeled D&C Yellow No. 11 was rapidl y 

absorbed, distributed, and excreted by male F344/ N 

rats (El Dareer et al., 1988). D&C Yellow No. 11 was 

not concentrated to any great extent in any tissu e 

(range 0.006% to 0.88% per tissue 72 hours following 

intravenous, oral, or repeated oral administration) and 

was rapidly excreted in feces following both oral and 

intravenous administration. O n a μg Eq/g tissue basis, 

the concentration of D&C Yellow No. 11-derive d 

radioactivity 72 hours after administration wa s 

approximately an order of magnitude greater in th e 

liver and kidney than in the other tissues. Excretion in 

the feces accounted for approxima tely 80% of an intravenous dose within 24 hours of administration an d 

85% within 72 hours. Most of the remainder of th e 

dose was detected in the urine within 24 hours wit h 

only trace amounts, approximately 2%, remaining i n 

the tissues 72 hours after dosing. Results of bil e 

cannulation studies indicated that excretion in feces is 

the result of rapid metabolism and excretion in bile (El 

Dareer et al., 1988). Greater than 50% of an intra ­

venous dose was excreted in bile within 4 hours o f 

administration. 

Humans 

No information on the absorption, distribution , 

metabolism, or excretion of D&C Yellow No. 11 i n 

humans was found in the literature. 

TOXICITY 

Experimental Animals 

No deaths occurred in albino rats (strain not given ) 

given 2,500 to 50,000 ppm D&C Yellow No. 11 i n 

feed for 13 weeks; however, the liver was enlarged at 

all concentrations studied (Hansen et al., 1960). 

In a study by Sun et al. (1987), F344/N rats wer e 

exposed to Solvent Yellow 33 aerosol by inhalatio n 

6 hours per day, 5 days per week, for 14 days o r 

13 weeks. After 14 days of exposure (10, 51, o r 3 3 230 mg/m ), rats exposed to 230 mg/m had body 

weights 8% lower than those of the controls. Afte r 3 13 weeks of exposure (1, 10.8, or 100 mg/m ), rats 3 exposed to 100 mg/m had body weights 5% lowe r

than those of the controls and an accumulation o f 

vacuolated alveolar macrophages in the lung . 

However, tissue analysis by high-performance liqui d 

chromatography showed very little Solvent Yellow 33 

in the lung after exposure, indicating rapid clearance. 

In toxicity studies conducted by the NTP, D& C 

Yellow No. 11 (approximately 99% pure) wa s 

administered in feed to male and female F344/N rat s 

and B6C3F1 mice at concentrations up to 50,000 ppm 

for 14 days (five animals per group) or 13 week s 

(10 animals per group) (NTP, 1991a). Although th e 

estimated intake of D&C Yellow No. 11 by mice was 

more than twice that by rats, the results of the 14-day 

and 13-week studies were s imilar for both species. No 

deaths occurred in rats or mice in the 14-day or 13 ­

week studies, but body weight gains were slightl y 

reduced in male and female rats given 17,000 o r 

50,000 ppm. Liver weights of exposed rats and mice 

were greater than those of controls. There was mini ­

mal to mild degeneration of the periportal portion o f 

the liver lobules of rats given 1, 700 ppm or greater and 

of mice given 5,000 ppm or greater. A dose-relate d 

yellow-brown pigment was observed in th e 

hepatocytes, Kupffer cells, and biliary epithelium i n 

male and female rats and mice and in the renal tubule 

epithelium in male and female rats. Hepatocellula r 

degeneration progressed slightly in severity wit h 

increased time of exposure (14 days versus 13 weeks) 

in rats but not in mice. Cytoplasmic alteration, a n 

increase in the size and number of hyaline droplets, in 

the renal tubule epithelium of the cortex and oute r 

medulla was present in all exposed groups of mal e 

rats. The conclusions from these studies were tha t 

D&C Yellow No. 11 caused increased liver weights in 

male and female rats and mice and increases in the size 

and number of hyaline droplets in male rats at al l 

exposure concentrations. 

Because of the cytoplasmic alteration (protein droplet 

accumulation) observed in male rats given D& C 

Yellow No. 11 in the 13-week NTP study (NTP , 

1991a), additional studies were co nducted to determine 

the potential for regression of these chemical-relate d 

lesions (Eastin et al., 1996). Groups of six male rats 

given feed containing 5,000 ppm D&C Yellow No. 11 

or untreated feed for 70 days, then maintained o n 

undosed feed, were examined the last day of exposure 

(day 1) and on days 3, 14, and 28 of the recover y 
D&C Yellow No. 11, NTP TR 463 15 

period. On day 1, cytoplasmic alteration and pigment 

in the renal tubules and hepatocellular degeneratio n 

and pigmentation were similar to the lesions observed 

at the same exposure concentration in the 13-wee k 

study. After a recovery period of 3 days, the severities 

of cytoplasmic alteration and pigmentation in the renal 

tubule epithelium were reduced in all rats. At thi s 

time, there was no longer morphologic evidence of the 

hepatocellular degeneration, and although th e 

pigmentation was slightly less prominent, it was stil l 

present in the biliary epithelium and cytoplasm o f 

hepatocytes and Kupffer cells in the periportal areas . 

After recovery periods of 14 or 28 days, pigment was 

still present in the renal tubule epithelium and live r 

biliary epithelium of all exposed rats. Ultrastructural 

features included an electron-dense, homogenou s 

pigment in the cytoplasm of canaliculi, bile duc t 

epithelium, and the lumen of bile ducts. Protei n 

droplet accumulation resembled 2μ-globulin by light 

microscopy; however, there was no evidence of a n 

increase in the amount of 2μ-globulin (as percent of 

total protein) measured by an ELISA metho d 

(Charbonneau et al., 1987; Yuan et al., 1992) in the 

kidney of rats with cytoplasmic alteration. Whe n 

measured on day 1, the amount of 2μ-globulin in the 

kidney of control and exposed rats was 10.0% an d 

8.1%, respectively. On days 3, 14, and 28, thes e 

values were similar in exposed and control groups. 

When partially hepatectomized Charle s River male rats 

were given 15,000 ppm D&C Yellow No. 11 in fee d 

for 10 days after surgery, liver regeneration wa s 

stimulated significantly compared with that in partially 

hepatectomized controls (Gershbein, 1982). 

D&C Yellow No. 11 was shown to sensitize the ski n 

of adult Hartley guinea pigs. Females induced wit h 

40% D&C Yellow No. 11 in ethanol with a 24-hou r 

occluded patch once a week for 3 consecutive week s 

responded to a challenge concentration of 10% administered after a 2-week rest period (Lamson et al., 

1982). Hartley guinea pigs were also induced by injection of emulsified Freund’s complete adjuvant into the 

nuchal region, followed by application of one of fou r 

test samples of D&C Yellow No. 11 to abraded ski n 

for 2 days and topical application on days 8 and 9 

(Sato et al., 1984). Challenge was carried out b y 

topical application on day 21 to flank skin. In thes e 

studies, the threshold concentration for induction and 

challenge was 10 ppm. In another study, D&C Yellow 

No. 11 in Freund’s adjuvant injected into the footpad 

produced dose-response hypersensitivity in femal e 

Hartley guinea pigs 2 weeks after exposure t o 

intradermal challenges of the dye (Palazzolo an d 

DiPasquale, 1983). Histopathologic examination o f 

reaction sites indicated a cellular inflammator y 

response in guinea pigs consistent with delayed-typ e 

hypersensitivity. 

Humans 

D&C Yellow No. 11 has been shown to have a hig h 

allergenic potential in humans (Kita et al., 1984). 

Patients sensitized to D&C Yellow No. 11 i n 

maximization tests exhibited an allergic contac t 

dermatitis from the use of soaps (Jordan, 1981 ; 

Weaver, 1983) and facial cosmetics (Björkner an d 

Magnusson, 1981; Calnan, 1981; Björkner an d 

Niklasson, 1983; Rapaport, 1984) containing this dye. 

Positive reactions were seen in beauticians with hand 

dermatitis given Quinoline Yellow SS (0.5% i n 

petrolatum) (Matsunaga et al., 1988). 

REPRODUCTIVE AND 

DEVELOPMENTAL TOXICITY 

Experimental Animals 

In a perinatal exposure study, body weight gains of rat 

dams given diets containing 5,000, 17,000, o r 

50,000 ppm D&C Yellow No. 11 for 4 weeks before 

mating to unexposed males were similar to that of the 

controls at the time of mating but were lower tha n 

those of the controls at parturition and at weanin g 

(NTP, 1991a). However, fertility, length of gestation, 

litter size, and pup birth body weights were unaffected 

by exposure. At weaning, body weights of pups from 

all exposed dams were lower than those from contro l 

dams. After potential exposure to D&C Yello w 

No. 11 for 4 weeks through the milk and subsequently 

in feed at the same concentrations given their dams , 

body weights of male and female pups give n 

5,000 ppm were similar to those of the controls, bu t 

those of pups given 17,000 or 50,000 ppm were lower 

than those of the controls. Microscopic evaluatio n 

showed lesions in pups from all exposure groups ; 

these lesions were similar to those in the liver an d 

kidney of rats in the 14-day and 13-week studie s 

(NTP, 1991a). In the liver, degeneration of hepato ­

cytes was present in all exposed groups and wa s 

characterized by minimal cytoplasmic vacuolization . 
16 D&C Yellow No. 11, NTP TR 463 

All exposed rats had a minimal accumulation of a 

granular-to-globular yellow-brown pigment in th e 

cytoplasms of cells in the liver and kidney. In th e 

kidney of exposed males, there was cytoplasmi c 

alteration (hyaline droplets) similar to that observed in 

the males in the 14-day and 13-week studies (NTP , 

1991a; Eastin et al., 1996). 

Humans 

No information on the reproductive or developmental 

toxicity of D&C Yellow No. 11 in humans was found 

in the literature. 

CARCINOGENICITY 

No information on the carcinogenic potential of D&C 

Yellow No. 11 in experimental animals or in humans 

was found in the literature. 

GENETIC TOXICITY 

D&C Yellow No. 11 has been shown to be mutagenic 

in vitro. It induced mutations in Salmonella 

typhimurium strains TA98 and TA100 when exposure 

occurred in the presence of S9 metabolic activatio n 

enzymes (Table C1; Zeiger et al., 1988). In a second 

study, mutations were induced in S. typhimurium 

strains TA102 and TA104 with and without S 9 

(Moore et al., 1988). D&C Yellow No. 11 was als o 

mutagenic and clastogenic to L5178Y/TK mous e 

lymphoma cells with and without S9 (Meyer et al., 

1986; Moore et al., 1988). Sister chromatid exchange 

levels were also elevated in mouse lymphoma cell s 

treated with D&C Yellow No. 11 in the presence of S9 

(Moore et al., 1988). In contrast to the demonstrated 

in vitro mutagenicity of D&C Yellow No. 11 in a 

number of assays, no increase in the frequency of 

sister chromatid exchanges was observed in vivo in 

bone marrow cells of male mice administered a single 

intraperitoneal injection of 10, 20, or 40 mg D& C 

Yellow No. 11/kg body weight (Moore et al., 1988). 

STUDY RATIONALE 

D&C Yellow No. 11 was nominated to the NTP fo r 

toxicity and carcinogenesis studies as part of a larger 

regulatory effort mandated by Congress an d 

undertaken by the FDA to determine the safety of a 

number of provisionally listed dyes. Currently, D&C 

Yellow No. 11 is regulated by the FDA for externa l 

use only (21 CFR, §74.1711). The decision to obtain 

toxicity and carcinogenesis data for this color additive 

by dietary exposure studies was based on the fact that 

it is a contaminant of D&C Yellow No. 10, a colo r 

additive approved for internal use and a candidate for 

permanent listing. The toxic effects of oral exposure 

to D&C Yellow No. 11 were unk nown in mice and had 

not been determined in rats in 2-year studies. Thus , 

14-day and 13-week toxicity s tudies were conducted in 

F344/N rats and B6C3F mice (NTP, 1991a) in order 1 

to compare the results to the NTP historical database 

for these strains. These studies were reported at th e 

time of their completion to the National Toxicolog y 

Program’s Board of Scientific Counselors’ Technical 

Reports Review Subcommittee. However, the FD A 

only requires carcinogenesis study data from on e 

rodent species, and different rat strains have been used 

the most, primarily the Sprague-Dawley rat. NTP also 

selected the rat based on the fact that the toxic effects 

in rats and mice after 13 weeks of dietary exposure to 

D&C Yellow No. 11 were basically the same, and rats 

were slightly more sensitive than mice. NTP also used 

the F344/N rat to be able to compare the results of the 

carcinogenicity study with their large historica l 

database on this strain. 
17 

MATERIALS AND METHODS
 

PROCUREMENT AND 

CHARACTERIZATION 

OF D&C YELLOW NO. 11 

D&C Yellow No. 11 was obtained fro m 

H. Kohnstamm and Company, Inc. (New York), in one 

lot (ZB2016) and certified by the Food and Dru g 

Administration, Division of Color Technology . 

Identity, purity, and stability analyses were conducted 

by the analytical chemistry laboratory, Midwes t 

Research Institute (Kansas City, MO) (Appendix G). 

Reports on analyses performed in support of the D&C 

Yellow No. 11 studies are on file at the Nationa l 

Institute of Environmental Health Sciences (NIEHS). 

The chemical, a yellow powder, w as identified as D&C 

Yellow No. 11 by infrared, ultraviolet/visible, nuclear 

magnetic resonance, and direct inlet mas s 

spectrometry. Purity was determined by elementa l 

analyses, Karl Fischer water analysis, thin-laye r 

chromatography, and high-perfo rmance liquid chromatography. Elemental analyses for carbon, hydrogen , 

and nitrogen were in agreement with the theoretica l 

values for D&C Yellow No. 11. Karl Fischer wate r 

analysis indicated less than 0.02% water. Thin-laye r 

chromatography indicated one major spot by on e 

system and one major spot and one trace impurity by 

a second system. High-performance liqui d 

chromatography revealed a major peak and tw o 

impurities with areas greater than 0.1% of the majo r 

peak area. The overall purity was determined to b e 

approximately 99%. 

Stability studies of the bulk chemical were performed 

using high-performance liquid ch romatography. These 

studies indicated that D&C Yellow No. 11 was stable 

as a bulk chemical for 2 weeks when stored protected 

from light at temperatures up to 60 C. To ensure 

stability, the bulk chemical was stored at roo m 

temperature in its original packaging protected fro m 

light. Stability was monitored during the reproductive 

toxicity and 2-year studies using high-performanc e 

liquid chromatography. No degradation of the bul k 

chemical was detected. 

PREPARATION AND ANALYSIS 

OF DOSE FORMULATIONS 

The dose formulations were prepa red every 2 weeks by 

mixing D&C Yellow No. 11 with feed (Table G1) . 

Homogeneity and stability studies of the 500 ppm dose 

formulation were performed by the analytica l 

chemistry laboratory using high-performance liqui d 

chromatography. Homogeneity was confirmed and the 

stability of the dose formulations was confirmed for at 

least 3 weeks when stored pro tected from light at room 

temperature and for 7 days when stored open to air and 

light. 

Periodic analyses of the dose formulations of D& C 

Yellow No. 11 were conducted at the study laboratory 

using visible spectrometry. During the reproductiv e 

toxicity and 2-year studies, the formulations wer e 

analyzed approximately every 8 weeks (Table G2) . 

All of the dose formulations used in the studies wer e 

within 10% of the target concentration. Due to a n 

unacceptable ratio of duplicate analyses, one dos e 

formulation was remixed. Res ults of a referee analysis 

performed by the analytical chemistry laborator y 

agreed with the results obtained by the stud y 

laboratory (Table G3). 

REPRODUCTIVE TOXICITY STUDY 

The reproductive toxicity study was conducted t o 

evaluate the cumulative toxic effects of parental an d 

in utero exposure to D&C Yellow No. 11; pups from 

this study continued to receive dosed feed at the same 

concentrations as their dams for the 2-year study. 

Thirty-two-day-old male and female F344/N first ­

generation (F ) rats were obtained from Taconic Farms 0 

(Germantown, NY) and quarantined for 10 days before 

receiving test diets. Rats were 112 days old on th e 

first day of cohabitation. Before initiation of the study, 

five male and five female rats were randomly selected 

for parasite evaluation and gross observation fo r 

evidence of disease. At the end of the study, serologic 

analyses were performed on five male and five female 
18 D&C Yellow No. 11, NTP TR 463 

control rats using the protocols of the NTP Sentine l 

Animal Program (Appendix J). 

Groups of 60 male and 60 female rats were fed diet s 

containing 0, 500, 1,700, or 5,000 ppm D&C Yellow 

No. 11 beginning 10 weeks prior to cohabitation , 

during cohabitation, and through gestation an d 

lactation (females). Feed and water were availabl e 

ad libitum. Rats were housed five per cage except 

during cohabitation (one male and one female pe r 

cage) and lactation (one dam and litter per cage) . 

Clinical findings, feed consumption, and body weights 

were recorded on day 1 (feed consumption day 2), once 

per week before cohabitation, on days 0, 6, 15, and 21 

of gestation (females), and on days 1, 4, 14, and 21 of 

lactation (females and pups). Details of the stud y 

design and animal maintenance are summarized i n 

Table 1; the following timeline describes the exposure 

periods. 

First-generation (F 0 ) rats started on D&C Yellow No. 11 in feed

70-day precohabitation period 

7-day cohabitation period 

21-day gestation period 

28-day lactation period 

Second-generation (F 1 ) rats continued on same concentrations

of D&C Yellow No. 11 in feed as dams for 2 years 

Terminal sacrifice 

During cohabitation, vaginal smears were taken daily 

from females to determine the pre sence of sperm. Rats 

showing no signs of littering by day 25 were killed , 

and uteri were examined for evidence of unsuccessful 

pregnancy. If there was no gross evidence o f 

pregnancy, uteri were stained with ammonium sulfide 

or sodium sulfide and examined for implantation sites. 

After parturition, clinical signs and number 

and sex of live pups were recorded. On day 4 

postpartum, litters were randomly culled to a 

maximum of eight pups (four male and four female ) 

per litter; on day 21, 60 male and 60 female pups were 

randomly selected from the litters of each exposur e 

group, and these pups were weaned on day 28 an d 

continued on the same test diet as their dams. Clinical 

findings and pup weights were recorded on days 1, 4, 

14, and 21. 
19 

1 

D&C Yellow No. 11, NTP TR 463 

2-YEAR STUDY 

Study Design 

Groups of 60 male and 60 female second-generatio n 

(F ) rats were fed diets containing 0, 500, 1,700, o r

5,000 ppm D&C Yellow No. 11 f or 105 to 106 weeks. 

Up to 10 male and 10 female rats from each grou p 

were evaluated at 12 months for hematology, orga n 

weights, and histopathology. 

Source and Specification of Animals 

Male and female F1 rats were selected from litter s 

produced by breeding male and female F344/N rats inhouse after exposure to 0, 500, 1,700, or 5,000 pp m 

D&C Yellow No. 11 in feed for 70 days. Rats wer e 

monitored for parasites throughout the study. Rat s 

were 28 days old when weaned at the beginning of the 

study. The health of the animal s was monitored during 

the studies according to the protocols of the NT P 

Sentinel Animal Program (Appendix J). 

Animal Maintenance 

Rats were housed five per cage. Feed and water were 

available ad libitum. Feed consumption was measured 

during week 2 and at monthly intervals thereafter b y 

cage (Appendix H). Cages and racks were rotate d 

every 2 weeks. Further details of animal maintenance 

are given in Table 1. Information on feed composition 

and contaminants is provided in Appendix I. 

Clinical Examinations and Pathology 

All animals were observed twice daily. Clinical findings and body weights were recorded at least once a 

week for the first 13 weeks and every 4 week s 

thereafter. 

A complete necropsy and microscopic examinatio n 

were performed on all rats. At the 12-month interi m 

evaluation, the liver and right kidn ey were weighed. At 

necropsy, all organs and tissues were examined fo r 

grossly visible lesions, and all major tissues were fixed 

and preserved in 10% neutral buffered formalin , 

processed and trimmed, embedded in paraffin , 

sectioned to a thickness of approximately 5 μm, an d 

stained with hematoxylin and eosin for microscopi c 

examination. For all paired organs (i.e., adrenal gland, 

kidney, ovary), samples from each organ wer e 

examined. For extended evaluation of rena l 

proliferative lesions, kidneys were step sectioned a t 

1 mm intervals, and four additional sections wer e 

obtained from each kidney. Tissues examine d 

microscopically are listed in Table 1. 

Hematology studies were performed on up to 10 male 

and 10 female rats per group at the 12-month interim 

evaluation. Rats were anesthetized with a CO 2 /O2

mixture, and blood was drawn from the retroorbita l 

sinus. Blood for hematology determinations wa s 

placed in tubes containing potassiu m 

ethylenediaminetetraacetic acid as an anticoagulant. 

The hematology variables evaluated are listed i n 

Table 1. Erythrocyte and leukocyte counts , 

hemoglobin concentration, hematocrit, mean cel l 

volume, mean cell hemoglobin, mean cell hemoglobin 

concentration, and platelet counts were performed on 

a Technicon H-1 hematology analyzer (Tarrytown , 

NY). Differential leukocyte counts, morphologi c 

evaluation of blood cells, and nucleated erythrocyt e 

counts were determined by light microscopy usin g 

smears prepared from blood stained by incubatin g 

equal volumes of whole blood and new methylene blue 

for at least 20 minutes. 

Microscopic evaluations were completed by the study 

laboratory pathologist, and the pathology data wer e 

entered into the Toxicology Data Management System. 

The microscopic slides, paraffin blocks, and residua l 

wet tissues were sent to the NTP Archives fo r 

inventory, slide/block match, and wet tissue audit. The 

slides, individual animal data records, and patholog y 

tables were evaluated by an independent qualit y 

assessment laboratory. The individual animal records 

and tables were compared for accuracy, the slide an d 

tissue counts were verified, and the histotechnique was 

evaluated. For the 2-year study, a quality assessment 

pathologist reviewed the forestomach, kidney, liver , 

lung, lymph nodes, salivary glands, and spleen o f 

males and females; the mammary gland, oral mucosa, 

pancreas, parathyroid gland, small intestine, an d 

tongue of males; and the clitoral gland of females. 

The quality assessment report and the reviewed slides 

were submitted to the NTP Pathology Working Group 

(PWG) chairperson, who reviewed the selected tissues 

and addressed any inconsistencies in the diagnose s 

made by the laboratory and quality assessment path ­

ologists. Representative histopathology slides con ­

taining examples of lesions related to chemica l 

administration, examples of disagreements in diag ­

noses between the laboratory and quality assessmen t 
20 D&C Yellow No. 11, NTP TR 463 

pathologist, or lesions of general interest wer e 

presented by the chairperson to the PWG for review . 

The PWG consisted of the quality assessment pathologist and other pathologists experienced in roden t 

toxicologic pathology. This group examined th e 

tissues without any knowledge of dose groups o r 

previously rendered diagnoses. When the PWG consensus differed from the opinion of the laborator y 

pathologist, the diagnosis was changed. Thus, th e 

final diagnoses represent a consensus of qualit y 

assessment pathologists, the PWG chairperson, an d 

the PWG. Details of these review procedures hav e 

been described, in part, by Maronpot and Boorma n 

(1982) and Boorman et al. (1985). For subsequent 

analyses of the pathology data, the diagnosed lesion s 

for each tissue type were evaluated separately o r 

combined according to the guidelines of McConnel l 

et al. (1986). 

STATISTICAL METHODS 

Survival Analyses 

The probability of survival was estimated by th e 

product-limit procedure of Kaplan and Meier (1958 ) 

and is presented in the form of graphs. Animals found 

dead of other than natural causes were censored from 

the survival analyses; animals dying from natura l 

causes were not censored. Statistical analyses fo r 

possible dose-related effects on survival used Cox’ s 

(1972) method for testing two groups for equality and 

Tarone’s (1975) life table test to identify dose-related 

trends. All reported P values for the survival analyses 

are two sided. 

Calculation of Incidence 

The incidences of neoplasms or nonneoplastic lesions 

as presented in Tables A1, A5, B1, and B5 are given 

as the number of animals bearing such lesions at a 

specific anatomic site and the number of animals with 

that site examined microscopically. For calculation of 

statistical significance, the incidences of mos t 

neoplasms (Tables A3 and B3) and all nonneoplastic 

lesions are given as the numbers of animals affected at 

each site examined microscopically. However, whe n 

macroscopic examination was required to detec t 

neoplasms in certain tissues (e.g., harderian gland , 

intestine, mammary gland, oral ca vity, and skin) before 

microscopic evaluation, or when neoplasms ha d 

multiple potential sites of occurrence (e.g., leukemia or 

lymphoma), the denominators consist of the number of 

animals on which a necropsy was performed. Tables 

A3 and B3 also give the survival-adjusted neoplas m 

rate for each group and each site-specific neoplasm , 

i.e., the Kaplan-Meier estimate of the neoplas m 

incidence that would have been observed at the end of 

the study in the absence of mortality from all othe r 

competing risks (Kaplan and Meier, 1958). 

Analysis of Neoplasm Incidences 

The majority of neoplasms in these studies wer e 

considered to be incidental to the cause of death or not 

rapidly lethal. Thus, the primary statistical metho d 

used was logistic regression analysis, which assumed 

that the diagnosed neoplasms were discovered as th e 

result of death from an unrelated cause and thus di d 

not affect the risk of death. In this approach, neoplasm 

prevalence was modeled as a logistic function o f 

chemical exposure and time. Bo th linear and quadratic 

terms in time were incorporated initially, and th e 

quadratic term was eliminated if the fit of the mode l 

was not significantly enhanced. The neoplas m 

incidences of exposed and control groups wer e 

compared on the basis of the likelihood score test for 

the regression coefficient of dose. This method o f 

adjusting for intercurrent mortality is the prevalenc e 

analysis of Dinse and Lagakos (1983), furthe r 

described and illustrated by Dinse and Hasema n 

(1986). When neoplasms are incidental, thi s 

comparison of the time-specific neoplasm prevalences 

also provides a comparison of the time-specifi c 

neoplasm incidences (McKnight and Crowley, 1984). 

In addition to logistic regression, other methods o f 

statistical analysis were used, and the results of these 

tests are summarized in the appendixes. Thes e 

methods include the life table test (Cox, 1972; Tarone, 

1975), appropriate for rapidly lethal neoplasms, an d 

the Fisher exact test and the Cochran-Armitage trend 

test (Armitage, 1971; Gart et al., 1979), procedures 

based on the overall proportion of neoplasm-bearin g 

animals. 

Tests of significance included pairwise comparisons of 

each exposed group with controls and a test for a n 

overall dose-related trend. Continuity-corrected tests 

were used in the analysis of neoplasm incidence, an d 

reported P values are one sided. The procedure s 

described in the preceding paragraphs were also used 
D&C Yellow No. 11, NTP TR 463 21 

to evaluate selected nonneoplastic lesions. For further 

discussion of these statistical methods, refer t o 

Haseman (1984). 

Analysis of Nonneoplastic Lesion Incidences 

Because all nonneoplastic lesions in this study wer e 

considered to be incidental to the cause of death or not 

rapidly lethal, the primary statistical analysis used was 

a logistic regression analysis in which nonneoplasti c 

lesion prevalence was modeled as a logistic function of 

chemical exposure and time. For lesions detected a t 

the interim evaluation, the Fisher exact test, a 

procedure based on the overall proportion of affected 

animals, was used. 

Analysis of Continuous Variables 

Two approaches were employed to assess the significance of pairwise comparisons between exposed an d 

control groups in the analysis of continuous variables. 

Organ and body weight data, which have approxi ­

mately normal distributions, were analyzed using th e 

parametric multiple comparis on procedures of Dunnett 

(1955) and Williams (1971, 1972). Hematology data, 

which have typically skewed distributions, wer e 

analyzed using the nonparametric multiple comparison 

methods of Shirley (1977) and Dunn (1964) . 

Jonckheere’s test (Jonckheere, 1954) was used t o 

assess the significance of the dose -related trends and to 

determine whether a trend-sensitive test (Williams’ or 

Shirley’s test) was more appropriate for pairwise comparisons than a test that does not assume a monotonic 

dose-related trend (Dunnett’s or Dunn’s test). Prior to 

analysis, extreme values identifi ed by the outlier test of 

Dixon and Massey (1951) were examined by NT P 

personnel, and implausible values were eliminate d 

from the analysis. Average severity values wer e 

analyzed for significance using the Mann-Whitney U 

test (Hollander and Wolfe, 1973). 

Reproductive Toxicity Data 

Body weight data for F rats, maternal body weigh t 0 

data during gestation and lactation, litter weight data, 

pup delivery data, percent male pups, and pups surviving on days 4 and 21 were analyzed using Williams’ 

or Dunnett’s test. Feed consumption data for F 0 rats 

were analyzed using Dunn’s or Shirley’s test. 

Historical Control Data 

Although the concurrent control group is always th e 

first and most appropriate control group used fo r 

evaluation, historical control data can be helpful in the 

overall assessment of neoplasm incidence in certai n 

instances. Consequently, neoplasm incidences fro m 

the NTP historical control database which is update d 

yearly are included in the NTP reports for neoplasms 

appearing to show compound-related effects. 

QUALITY ASSURANCE METHODS 

The reproductive toxicity and 2-year studies wer e 

conducted in compliance with Food and Dru g 

Administration Good Laboratory Practice Regulations 

(21 CFR, Part 58). In addition, as records from th e 

2-year study were submitted to the NTP Archives, this 

study was audited retrospectively by an independen t 

quality assurance contractor. Separate audits covering 

completeness and accuracy of the pathology data , 

pathology specimens, final pathology tables, and a 

draft of this NTP Technical Report were conducted . 

Audit procedures and findings are presented in th e 

reports and are on file at NIEHS. The audit finding s 

were reviewed and assessed by NTP staff, so al l 

comments had been resolved or were otherwis e 

addressed during the preparation of this Technica l 

Report. 

GENETIC TOXICOLOGY 

The genetic toxicity of D&C Yellow No. 11 wa s 

assessed by testing the ability of the chemical to induce 

mutations in various strains of Salmonella 

typhimurium, sister chromatid exchanges an d 

chromosomal aberrations in cultured Chinese hamster 

ovary cells, and increases in the frequency of micronucleated erythrocytes in mouse peripheral blood. Th e 

protocols for these studies and the results are given in 

Appendix C. 

The genetic toxicity studies of D&C Yellow No. 11 are 

part of a larger effort by the NTP to develop a 

database that would permit the evaluation o f 

carcinogenicity in experimental animals from th e 

structure and responses of the chemical in short-term 
22 D&C Yellow No. 11, NTP TR 463 

in vitro and in vivo genetic toxicity tests. Thes e Other in vitro genetic toxicity tests do not correlat e 

genetic toxicity tests were originally developed t o well with rodent carcinogenicity (Tennant et al., 1987; 

study mechanisms of chemically in duced DNA damage Zeiger et al., 1990), although these other tests ca n 

and to predict carcinogenicity in animals, based on the provide information on the types of DNA an d 

electrophilic theory of chemical carcinogenesis and the chromosome effects that can be induced by th e 

somatic mutation theory (Miller and Miller, 1977 ; chemical being investigated. Data from NTP studie s 

Straus, 1981; Crawford, 1985). show that a positive response in Salmonella is 

currently the most predictive in vitro test for rodent 

There is a strong correlation between a chemical’ s carcinogenicity (89% of the Salmonella mutagens 

potential electrophilicity (structural alert to DN A were rodent carcinogens), and that there is n o 

reactivity), mutagenicity in Salmonella, and complementarity among the in vitro genetic toxicity 

carcinogenicity in rodents. The combination o f tests. That is, no battery of tests that included th e 

electrophilicity and Salmonella mutagenicity is highly Salmonella test improved the predictivity of th e 

correlated with the induction of carcinogenicity in rats Salmonella test alone. The predictivity for carcino ­

and mice and/or at multiple tissue sites (Ashby an d genicity of a positive response in bone marro w 

Tennant, 1991). chromosome aberration or micronucleus tests is not yet 

defined. 
D&C Yellow No. 11, NTP TR 463 23 

TABLE 1 

Experimental Design and Materials and Methods in the Feed Studies of D&C Yellow No. 11 

Reproductive Toxicity Study 2-Year Study 

Study Laboratory 

Southern Research Institute 

(Birmingham, AL) and Argus Research Laboratories, Inc. 

(Horsham, PA) 

Southern Research Institute 

(Birmingham, AL) 

Strain and Species 

F344/N rats F344/N rats 

Animal Source 

Taconic Farms 

(Germantown, NY) 

Bred in-house 

Time Held Before Studies 

10 days Not applicable 

Average Age When Studies Began 

42 days 28 days at weaning 

Date of First Dose 

18 December 1989 26 April 1990 

Duration of Dosing 

Males: 13 weeks 

Females: 19 weeks 

Males: 105 weeks 

Females: 106 weeks 

Date of Last Dose 

Males: 13 March 1990 

Females: 24 April 1990 

12-Month interim evaluation — 

males: 17 April 1991 

females: 18 April 1991 

Terminal — 

males: 27 April 1992 

females: 4 May 1992 

Necropsy Dates 

Not applicable 12-Month interim evaluation — 

males: 17 April 1991 

females: 18 April 1991 

Terminal — 

males: 27-28 April 1992 

females: 4-6 May 1992 

Average Age at Necropsy 

Not applicable 12-Month interim evaluation — 

males: 56 weeks 

females: 56 weeks 

Terminal — 

males: 110 weeks 

females: 111 weeks 
24 D&C Yellow No. 11, NTP TR 463 

TABLE 1
 

Experimental Design and Materials and Methods in the Feed Studies of D&C Yellow No. 11 (continued)
 

Reproductive Toxicity Study	 2-Year Study 

Size of Study Groups 

60 males and 60 females 12-Month interim evaluation) 

6 to 10 males and 9 to 10 females 

Terminal )50 to 54 males and 50 to 51 females 

Method of Distribution 

Rats were distributed randomly into groups of approximately equal	 Litters culled twice using a table of random numbers to no more than 

initial mean body weights.	 four males and four females per litter on day 4, then two male and 

two female pups from each litter on day 21; 60 male and 60 female 

pups per exposure group were continued on study after weaning. 

Animals per Cage 

Before cohabitation: 5 5 

During cohabitation: 1 pair 

After cohabitation: 5 males 

or 1 dam and litter 

Method of Animal Identification 

Tail tattoo	 Tail tattoo 

Diet 

NIH-07 open formula mash (Zeigler Brothers, Inc., Gardners, PA), Same as reproductive toxicity study 

available ad libitum, changed weekly 

Water Distribution 

Tap water (Birmingham municipal supply) via automatic watering Same as reproductive toxicity study 

system (Edstrom Industries, Inc., Waterford, WI), available 

ad libitum 

Cages 

Solid-bottom polycarbonate (Lab Products, Maywood, NJ), changed Solid-bottom polycarbonate (Lab Products, Maywood, NJ), changed 

twice weekly except from day 18 of gestation through delivery twice weekly or when excessively soiled or wet 

Bedding 

Sani-Chips (P.J. Murphy Forest Products Corp., Montville, NJ), Sani-Chips (P.J. Murphy Forest Products Corp., Montville, NJ), 

changed twice weekly except from day 18 of gestation through changed twice weekly 

delivery 

Rack Filters 

Reemay® spun-bonded polyester (Andico, Birmingham, AL), Reemay® spun-bonded polyester (Andico, Birmingham, AL), 

changed once every 2 weeks except from day 18 of gestation through changed once every 2 weeks 

delivery 

Racks 

Stainless steel (Lab Products, Inc., Maywood, NJ), changed once Stainless steel (Lab Products, Inc., Maywood, NJ), changed once 

every 2 weeks except from day 18 of gestation through delivery every 2 weeks 
D&C Yellow No. 11, NTP TR 463 25 

TABLE 1
 

Experimental Design and Materials and Methods in the Feed Studies of D&C Yellow No. 11 (continued)
 

Reproductive Toxicity Study	 2-Year Study 

Animal Room Environment 

Temperature: 20.0 to 25.6 C Same as reproductive toxicity study 

Relative humidity: 23.3% to 81.2% 

Fluorescent light: 12 hours/day 

Room air: minimum of 10 changes per hour 

Doses 

0, 500, 1,700, or 5,000 ppm in feed, availablead libitum	 0, 500, 1,700, or 5,000 ppm in feed, availablead libitum 

Type and Frequency of Observation 

Observed twice daily; clinical findings and body weights were Observed twice daily; animals were weighed and clinical findings 

recorded on day 1 and weekly before cohabitation for F0 males and were recorded initially, weekly for 13 weeks, monthly thereafter, and

females, on days 0, 6, 15, and 21 of gestation for F0 females, and on	 at the end of the studies. Feed consumption was recorded during

days 1, 4, 14, and 21 of lactation for F females and F pups. 1 Feed	 week 2 and approximately monthly thereafter by cage. 0 

consumption was recorded by cage weekly before cohabitation, on 

days 0, 6, 15, and 21 during gestation, and on days 1, 4, 14, and 21 

during lactation. 

Method of Sacrifice 

CO asphyxiation	 CO asphyxiation 2 2 

Necropsy 

None	 Necropsy performed on all animals. Organs weighed at the 

12-month interim evaluation were the liver and right kidney. 

Clinical Pathology 

None	 Blood was collected from the retroorbital sinus of all 12-month 

interim evaluation rats. 

Hematology: hematocrit; hemoglobin; erythrocyte, reticulocyte, and 

nucleated erythrocyte counts; mean cell volume; mean cell 

hemoglobin; mean cell hemoglobin concentration; platelet counts; 

and total leukocyte counts and differentials 

Histopathology 

None	 Complete histopathology was performed on all rats. In addition to 

gross lesions and tissue masses, the tissues examined included: 

adrenal gland, brain, clitoral gland, esophagus, femur with marrow 

and epiphysis, heart and aorta, large intestine (cecum, colon, and 

rectum), small intestine (duodenum, jejunum, and ileum), kidneys, 

liver, lungs and mainstem bronchi, lymph nodes (mandibular and 

mesenteric), mammary gland, nose, ovaries, pancreas, parathyroid 

gland, pituitary gland, preputial gland, prostate gland, salivary gland, 

skin, spleen, stomach (forestomach and glandular stomach), testes, 

thymus, thyroid gland, trachea, urinary bladder, and uterus. 
26 D&C Yellow No. 11, NTP TR 463 
27 

RESULTS
 

DOSE SELECTION RATIONALE 

The results of the 13-week rat study were used to select 

doses of 500, 1,700, and 5,000 ppm for the current 

F344/N rat study. In the 13-week feed study, rats were 

given 500, 1,700, 5,000, 17,000, or 50,000 ppm. There 

was no perinatal exposure, and animals were about 6 

weeks old when placed on dosed feed. Mean body 

weights of males and females were significantly reduced 

after 13 weeks of exposure to 17,000 and 50,000 ppm, 

and there was mild hepatocellular periportal degeneration 

in 7 males given 17,000 ppm, in all 10 given 50,000 

ppm, and in 2 females given 50,000 ppm. This lesion 

was minimal at doses of 1,700 and 5,000 ppm in males 

(4/4, 9/10) and females (2/2, 7/7) and in females at 

17,000 ppm (9/10) and was not observed in groups given 

500 ppm. In addition, a range-finding study was 

conducted in which female rats were given 5,000, 

17,000, or 50,000 ppm D&C Yellow No. 11 in feed for 

4 weeks before mating and during mating, gestation, and 

the first 4 weeks after having litters. Pups were weaned 

at week 4 and continued on the same feed as their dams 

for an additional 4 weeks. Litters would have been 

potentially exposed in utero, through lactation, and feed. 

There was no difference between study groups in 

reproductive performance. However, pup body weights 

in the 17,000 and 50,000 ppm groups were decreased at 

8 weeks of age. Microscopic evaluation showed that the 

liver lesions in exposed pups were similar to those 

described for the 13-week study. 

Following discussions with the FDA, the nominator, the 

NTP conducted studies of perinatal exposure followed by 

dietary exposure for 2 years after weaning in male and 

female F344/N rats to assess the toxicity and 

carcinogenicity of D&C Yellow No. 11. This study was 

chosen to generate data similar to those used by the FDA 

to regulate other color additives, and the results are 

presented in this Technical Report. 

REPRODUCTIVE TOXICITY STUDY 

All first-generation (F ) male and female rats survived 0 

until the end of the study. Prior to cohabitation, mean 

body weight gains of males (days 1 to 71) given 500, 

1,700, or 5,000 ppm and of females (days 1 to 66) given 

5,000 ppm were significantly lower than those of the 

controls (Table F1). The mean body weight gains of 

exposed females during gestation and lactation were 

generally similar to those of the controls (Table F3). 

Feed consumption by exposed groups of rats was 

generally similar to that by the control groups prior to 

cohabitation (Table F2). Dietary levels of 500, 1,700, 

and 5,000 ppm D&C Yellow No. 11 resulted in average 

daily doses of approximately 35, 120, and 350 mg D&C 

Yellow No. 11/kg body weight to males and 35, 120, and 

370 mg/kg to females. 

Prior to cohabitation, clinical findings attributed to D&C 

Yellow No. 11 exposure included yellow discoloration of 

the entire body or fur in all males and females given 

1,700 or 5,000 ppm and in all male s and seven females 

given 500 ppm. All rats given 1,700 or 5,000 pp m 

had urine-stained abdominal fur. Yellow discoloration 

of the fur was observed in all exposed female rat s 

during gestation and lactation. 

The duration of gestation (Table F3), the average litter 

size, the number of live pups on days 4 (precull) an d 

21, and the percent male pups for each exposure group 

(Table F4) were similar to those of the controls. The 

mean body weights of exposed litters were signifi ­

cantly less than those of the control litters on days 14 

and 21; this effect was considered to be related t o 

D&C Yellow No. 11 exposure. 
28 

c 

D&C Yellow No. 11, NTP TR 463 

2-YEAR STUDY 5,000 ppm was significantly less than that of th e 

Survival controls. Survival of 1,700 ppm females wa s 

Estimates of 2-year survival probabilities for second- significantly greater than that of the controls. Survival 

generation (F 1 ) male and female rats are shown i n of 500 ppm males and females and of 5,000 pp m 

Table 2 and in the Kaplan-Meier survival curve s females was similar to that of the controls. 

(Figure 1). Survival of males given 1,700 or 

TABLE 2 

Survival of Rats in the 2-Year Feed Study of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Male 

Animals initially in study 60 60 60 60 

12-Month interim evaluationa 10 9 9 6 

Moribund 29 29 41 49 

Natural deaths 2 2 1 3 

Other 0 0 1 0 

Animals surviving to study termination 19 20 8 2 

b Percent probability of survival at end of study 38 39 16 4 

Mean survival (days) c 625 614 595 567 

d Survival analysis P<0.001 P=0.974 P=0.013 P<0.001 

Female 

Animals initially in study 60 60 60 60 

12-Month interim evaluationa 10 9 10 9 

Moribund 25 23 12 25 

Natural deaths 3 2 1 3 

Animals surviving to study termination 22 26 37 23 

Percent probability of survival at end of study 44 51 74 45 

Mean survival (days) 637 638 654 631 

Survival analysis P=0.882 P=0.769N P=0.006N P=1.000N 

a Censored from survival analyses b Kaplan-Meier determinations based on the number of animals alive on the first day of terminal sacrifice
 

Mean of all deaths (uncensored, censored, and terminal sacrifice) d The result of the life table trend test (Tarone, 1975) is in the control column, and the results of the life table pairwise comparisons (Cox, 1972) with 

the controls are in the exposure columns. A lower mortality in an exposure group is indicated byN. 
D&C Yellow No. 11, NTP TR 463 29 

FIGURE 1
 

Kaplan-Meier Survival Curves for Male and Female Rats Administered D&C Yellow No. 11
 

in Feed for 2 Years
 
30 D&C Yellow No. 11, NTP TR 463 

Body Weights, Feed and Compound 

Consumption, and Clinical Findings 

Male and female F1 rats were selected from litters i n 

the reproductive toxicity study; therefore , 

measurements of individual body weight data for F 1 

rats in the 2-year study began at weaning when the rats 

were 28 days old. Mean body weights of 1,700 an d 

5,000 ppm males and females were generally lowe r 

than those of the controls throughout the stud y 

(Figure 2; Tables 3 and 4). Final mean body weights 

of males were 95% (500 ppm), 93% (1,700 ppm), and 

85% (5,000 ppm) that of the controls, and those o f 

females were 99%, 95%, and 94% that of the controls. 

Feed consumption by exposed groups was similar t o 

that by the controls (Tables H1 and H2). Dietar y 

levels of 500, 1,700, and 5,000 ppm D&C Yellow No. 

11 resulted in average daily doses of approximatel y 

25, 85, and 250 mg D&C Yellow No. 11/kg bod y 

weight to males and 25, 100, and 280 mg/kg t o 

females. Chemical-related clinical findings include d 

yellow discoloration of the entire body in all exposed 

males and females from day 1 and head swelling an d 

edema in 1,700 and 5,000 ppm males . One 1,700 ppm 

male and five 5,000 ppm males were killed moribund

 between weeks 49 and 81; these deaths wer e 

attributed to extensive edema. 

Hematology 

A few minimal hematology differences occurred i n 

male rats at the 12-month interim evaluatio n 

(Table E1). There was evidence of minimal anemia in 

exposed males; this anemia was characterized b y 

decreased hematocrit values, hemoglobin concentra ­

tions, and erythrocyte counts. There were no differ ­

ences in the mean cell volume o r mean cell hemoglobin 

concentration in exposed rats, to indicate that erythrocytes were normocytic and normochromic. There were 

no increases in reticulocyte counts to indicate a bon e 

marrow response to the anemia. Therefore, th e 

minimal anemia was characterized as normocytic , 

normochromic, and nonresponsive. Normocytic , 

normochromic, nonresponsive anemias have bee n 

related to selective suppression of erythropoiesis in a 

variety of disorders and may be due to decrease d 

erythropoietin elaboration, bone marrow suppression, 

or defective iron metabolism. There were n o 

biologically or statistically significant differences i n 

hematology parameters between control and expose d 

females. 
D&C Yellow No. 11, NTP TR 463 29 

FIGURE 1
 

Kaplan-Meier Survival Curves for Male and Female Rats Administered D&C Yellow No. 11
 

in Feed for 2 Years
 
32 D&C Yellow No. 11, NTP TR 463 

TABLE 3 

Mean Body Weights and Survival of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 

Weeks 0 ppm 500 ppm 1,700 ppm 5,000 ppm 

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of 

Study (g) Survivors (g) controls) Survivors (g) controls) Survivors (g) controls) Survivors 

1 112 60 100 89 60 99 88 60 93 83 60 

2 159 60 145 91 60 142 89 60 133 84 60 

3 194 60 180 93 60 176 90 60 167 86 60 

4 227 60 209 92 60 204 90 60 197 87 60 

5 254 60 235 92 60 229 90 60 221 87 60 

6 277 60 260 94 60 253 91 60 240 87 60 

7 288 60 272 94 60 267 93 60 256 89 60 

8 303 60 289 96 60 284 94 60 271 90 60 

9 315 60 302 96 60 296 94 60 284 90 60 

10 327 60 315 96 60 310 95 60 297 91 60 

11 337 60 324 96 60 319 95 60 307 91 60 

12 347 60 336 97 60 331 96 60 317 91 60 

13 353 60 343 97 60 337 96 60 323 91 60 

17 383 60 371 97 60 365 96 60 352 92 60 

21 401 60 390 97 60 385 96 60 370 93 60 

25 416 60 405 97 60 400 96 60 384 92 60 

29 431 60 421 98 60 413 96 60 400 93 60 

33 425 60a 427 100 59a 423 100 60 404 95 60a 

37 449 60 436 97 59 432 96 59 416 93 60 

41 455 60 443 97 59 439 97 59 421 93 60 

45 456 60 449 98 59 443 97 59 423 93 59 

49 

53b 

464 

472 

60 

50 

454 

460 

98 

98 

59 

50 

450 

457 

97 

97 

59 

50 

430 

438 

93 

93 

58 

50 

57 472 50 461 98 50 460 97 49 438 93 49 

61 475 50 462 97 50 461 97 48 441 93 49 

65 477 50 463 97 50 460 96 47 439 92 48 

69 475 50 463 97 50 462 97 46 438 92 47 

73 472 48 460 98 48 456 97 43 436 93 39 

77 473 47 451 96 47 451 95 43 433 92 37 

81 470 47 446 95 42 442 94 42 425 91 35 

85 463 46 445 96 38 435 94 41 413 89 30 

89 452 42 439 97 36 433 96 36 404 89 25 

93 451 35 425 94 33 422 94 30 401 89 22 

97 446 30 425 95 28 420 94 23 387 87 19 

101 435 24 414 95 26 403 93 13 368 85 8 

Mean for weeks 

1-13 269 255 95 250 93 239 89 

14-52 431 422 98 417 97 400 93 

53-101 464 447 96 443 95 420 91 

a 

b The number of animals weighed for this week is less than the number of animals surviving. 

Interim evaluation occurred during week 51. 
D&C Yellow No. 11, NTP TR 463 33 

TABLE 4 

Mean Body Weights and Survival of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 

Weeks 0 ppm 500 ppm 1,700 ppm 5,000 ppm 

on Av. Wt. No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of Av. Wt. Wt. (% of No. of 

Study (g) Survivors (g) controls) Survivors (g) controls) Survivors (g) controls) Survivors 

1 100 60 89 90 60 88 89 60 85 86 60 

2 126 60 117 93 60 115 92 60 113 90 60 

3 141 60 132 93 60 130 92 60 129 92 60 

4 152 60 145 95 60 143 94 60 143 94 60 

5 163 60 154 95 60 150 92 60 151 93 60a 

6 168 60 161 96 60 156 93 60 155 92 60 

7 177 60 168 95 60 164 93 60 163 92 60 

8 180 60 173 96 60 167 93 60 168 93 60 

9 186 60 178 96 60 174 93 60 173 93 60 

10 187 60 179 96 60 173 92 60 174 93 60 

11 193 60 185 96 60 181 94 60 180 93 60 

12 196 60 189 96 60 183 93 60 184 93 60 

16 206 60 200 97 60 196 95 60 194 94 60 

21 212 60 205 97 60 201 95 60 197 93 60 

24 217 60 212 97 60 207 95 60 205 94 59 

28 227 60 218 96 60 215 95 60 211 93 59 

32 233 60 227 97 60 221 95 60 219 94 59 

36 238 60 231 97 60 222 93 60 223 94 59 

40 246 60 240 98 60 231 94 60 233 95 59 

44 255 60 248 97 59 238 93 60 240 94 59 

48 

52b 

265 

277 

60 

50 

258 

276 

97 

100 

59 

50 

251 

259 

95 

94 

60 

50 

250 

263 

94 

95 

59 

50 

56 290 50 285 99 50 273 94 50 275 95 50 

60 296 50 293 99 50 280 95 50 282 95 50 

64 307 50 303 99 50 289 94 50 291 95 48 

68 314 50 312 99 49 300 96 49 299 95 47 

72 320 49 317 99 49 304 95 49 307 96 47 

76 323 49 320 99 49 305 95 49 308 95 45 

80 331 48 323 98 47 310 94 48 313 95 45 

84 339 45 330 97 47 315 93 47 317 93 44 

88 342 41 330 97 45 320 94 45 319 93 42 

92 348 40 333 96 38 327 94 44 320 92 42 

96 357 35 342 96 35 331 93 41 332 93 36 

100 355 33 344 97 31 336 95 41 334 94 35 

104 354 28 349 99 27 337 95 37 333 94 27 

Mean for weeks 

1-13 164 156 95 152 93 152 93 

14-52 238 232 97 224 94 224 94 

53-104 329 322 98 310 94 310 94 

a 

b The number of animals weighed for this week is less than the number of animals surviving. 

Interim evaluation occurred during week 52. 
 

34 D&C Yellow No. 11, NTP TR 463 

Pathology and Statistical Analyses 

This section describes the statistically significant o r 

biologically noteworthy changes in the incidences o f 

mononuclear cell leukemia and neoplasms and/o r 

nonneoplastic lesions of the liver, kidney, oral cavit y 

(oral mucosa and tongue), testis, forestomach, smal l 

intestine, salivary gland, pancreas, lymph nodes , 

clitoral gland, and pituitary gland. Summaries of th e 

incidences of neoplasms and nonneoplastic lesions , 

individual animal tumor diagnoses, statistical analyses 

of primary neoplasms that occurred with an incidence 

of at least 5% in at least one animal group, and his ­

torical incidences for the neoplasms mentioned in this 

section are presented in Appendix A for male rats and 

Appendix B for female rats. 

Liver: At the 12-month interim evaluation, absolut e 

and relative liver weights of all exposed groups o f 

males and females were significant ly greater than those 

of the controls (Table D1). At 2 years, the incidences 

of hepatocellular adenoma in 5,000 ppm males and of 

hepatocellular adenoma or carcinoma (combined) i n 

5,000 ppm females were significantly greater tha n 

those in the controls (Tables 5, A3, and B3); thes e 

neoplasms occurred with significant exposure-related 

trends. The incidence of hepatocellular adenoma i n 

5,000 ppm males exceeded the historical range (0% to 

10%; Tables 5 and A4a) in untreated controls fro m 

NTP feed studies; the incidences of adenoma o r 

carcinoma (combined) in 1,700 and 5,000 pp m 

females also exceeded the historical control range (0% 

to 6%; Tables 5 and B4a). Hepatocellular adenomas 

were discrete masses with distinct borders tha t 

compressed and replaced adjacent hepatic parenchyma 

(Plate 1). Hepatic cords within adenomas typicall y 

were at sharp angles to the cords in the adjacen t 

normal hepatic parenchyma. Adenomas had loss o f 

normal lobular pattern and usually lacked central veins 

and portal areas. Cells within adenomas were ofte n 

somewhat pleomorphic and had altered stainin g 

patterns. The hepatocellular carcinoma (Plate 2) in a 

5,000 ppm female was a discrete lesion with markedly 

disturbed architecture (clumps of cells separated b y 

irregular, relatively wide spaces) and more cellula r 

atypia than the adenomas. 

At 12 months, the incidences of clear cell foci in 1,700 

and 5,000 ppm females were significantly greater than 

that in the controls (Tables 5 and B5). At 2 years, the 

incidences of mixed cell foci in exposed males and of 

clear cell foci in exposed males (except 500 ppm) and 

females were significantly greater than those in th e 

controls (Tables 5, A5, and B5). In 1,700 ppm males 

at 2 years, the incidence of eosinophilic foci wa s 

significantly greater than that of the controls. At 1 2 

months and 2 years, the incidences of basophilic foc i 

in 1,700 and 5,000 ppm females appeared to b e 

significantly less than those of the controls; however, 

basophilic foci may have been obscured by cytologi c 

alterations. Foci of hepatocellular alteration wer e 

discrete areas within the liver with a relatively normal 

lobular architecture but having altered stainin g 

characteristics (Plate 3). 

The incidences of cytologic alterations of hepatocytes 

in all exposed groups of males and females were significantly greater than those in the controls a t 

12 months and 2 years (Tables 5, A5, and B5), and the 

severities generally increased with increasing exposure 

concentration. Cytologic alterations of hepatocyte s 

consisted of increases in basophilia and granularity in 

the cytoplasm of hepatocytes that involved primaril y 

periportal hepatocytes in mildly affected cases whil e 

more severely affected livers had diffuse involvement. 

The increased basophilia and granularity of th e 

cytoplasm of hepatocytes in many exposed rat s 

probably obscured detection of basophilic foci. Th e 

incidences of bile duct pigmentation in all expose d 

groups of males and females a t 12 months and 2 years, 

of hepatocyte pigmentation in exposed males an d 

females at 12 months (except 500 ppm males) an d 

2 years, and of Kupffer cell pigmen tation in 5,000 ppm 

males and females at 12 months and in 1,700 an d 

5,000 ppm males and females at 2 years wer e 

significantly greater than those in the controls. Th e 

severities of bile duct pigmentation and hepatocyt e 

pigmentation generally increased with increasin g 

exposure concentration. Pigmentation was a minimal 

to moderate accumulation of a golden to green-brown 

granular material within the cytoplasm of hepatocytes, 

bile duct epithelium, or, less commonly, Kupffer cells 

(Plate 4). Special stains of pigment in the 14-da y 

study (NTP, 1991a) were negative for hemosiderin , 

bile, and lipofuscin. The incidences of bile duc t 

hyperplasia in 1,700 and 5,000 ppm females a t 

12 months and 2 years were significantly greater than 

in the controls; however, the incidences in expose d 

males were significantly less than in the controls a t 

2 years. 
D&C Yellow No. 11, NTP TR 463 35 

TABLE 5 

Incidences of Neoplasms and Nonneoplastic Lesions of the Liver in Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Male 

12-Month Interim Evaluation 

Number Examined Microscopically 

Basophilic Focusa 10 1

 9

 1

 9

 0

 6 

1 

Clear Cell Focus 0 0 1 1 

Eosinophilic Focus 0 2 0 0 

Mixed Cell Focus

Bile Duct, Hyperplasia

 0

 4 (1.3)b 

0

 0

 0

 0

 1 

1 (1.0) 

Bile Duct, Pigmentation 0 9** (1.0) 9** (1.0) 6** (1.3) 

Hepatocyte, Cytologic Alterations 0 8** (1.0) 9** (1.6) 6** (1.7) 

Hepatocyte, Pigmentation 0 1 (1.0) 8** (1.0) 6** (1.2) 

Kupffer Cell, Pigmentation 0 0 0 5** (1.0) 

2-Year Study 

Number Examined Microscopically 50 51 51 54 

Basophilic Focus 14 8 6 7 

Clear Cell Focus 9 15 15* 18** 

Eosinophilic Focus 7 5 14* 12 

Mixed Cell Focus 1 10** 9** 10** 

Bile Duct, Hyperplasia 49 (2.1) 26** (1.5) 18** (1.4) 32** (1.5) 

Bile Duct, Pigmentation 0 38** (1.2) 51** (1.9) 54** (2.2) 

Hepatocyte, Cytologic Alterations 0 20** (2.1) 44** (2.4) 42** (2.7) 

Hepatocyte, Pigmentation 0 22** (1.0) 45** (1.7) 51** (2.1) 

Kupffer Cell, Pigmentation 7 (2.4) 15 (2.0) 23** (1.9) 26** (1.8) 

Hepatocellular Adenomac 

Overall rated 

Adjusted ratee 

Terminal ratef 

1/50 (2%) 

5.3% 

1/19 (5%) 

2/51 (4%) 

7.9% 

1/20 (5%) 

1/51 (2%) 

2.6% 

0/8 (0%) 

7/54 (13%) 

75.0% 

1/2 (50%) 

First incidence (days) 

Logistic regression testg 

733 (T) 

P=0.001 

656 

P=0.487 

607 

P=0.757 

498 

P=0.008 

Female 

12-Month Interim Evaluation 

Number Examined Microscopically 10 9 10 9 

Basophilic Focus 7 2 2* 1* 

Clear Cell Focus 0 1 4* 4* 

Eosinophilic Focus 0 0 1 0 

Bile Duct, Hyperplasia 1 (1.0) 1 (1.0) 6* (1.0) 9** (1.4) 

Bile Duct, Pigmentation 0 9** (1.2) 7** (1.0) 9** (2.3) 

Hepatocyte, Cytologic Alterations 0 4* (1.0) 10** (1.5) 9** (2.1) 

Hepatocyte, Pigmentation 0 9** (1.0) 10** (1.3) 9** (2.6) 

Kupffer Cell, Pigmentation 0 0 0 9** (1.4) 

(continued) 
36 D&C Yellow No. 11, NTP TR 463 

TABLE 5 

Incidences of Neoplasms and Nonneoplastic Lesions of the Liver in Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 (continued) 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Female (continued) 

2-Year Study 

Number Examined Microscopically 50 51 50 51 

Basophilic Focus 32 26 11** 12** 

Clear Cell Focus 10 18* 29** 30** 

Eosinophilic Focus 10 9 14 16 

Mixed Cell Focus 12 19 20 16 

Bile Duct, Hyperplasia 14 (1.3) 10 (1.7) 27** (1.5) 33** (1.5) 

Bile Duct, Pigmentation 0 46** (1.3) 49** (2.0) 50** (2.4) 

Hepatocyte, Cytologic Alterations 0 11** (2.3) 31** (2.1) 40** (2.5) 

Hepatocyte, Pigmentation 0 34** (1.1) 44** (2.1) 50** (2.4) 

Kupffer Cell, Pigmentation 9 (2.1) 11 (2.4) 16* (1.8) 32** (1.9) 

Hepatocellular Adenoma 

Overall rate 0/50 (0%) 2/51 (4%) 5/50 (10%) 4/51 (8%) 

Adjusted rate 0.0% 6.4% 13.5% 15.7% 

Terminal rate 

First incidence (days) 

0/22 (0%) 

—h 1/26 (4%) 

645 

5/37 (14%) 

740 (T) 

3/23 (13%) 

720 

Logistic regression test P=0.100 P=0.241 P=0.095 P=0.068 

Hepatocellular Carcinoma 

Overall rate 0/50 (0%) 0/51 (0%) 0/50 (0%) 1/51 (2%) 

Hepatocellular Adenoma or Carcinomai 

Overall rate 0/50 (0%) 2/51 (4%) 5/50 (10%) 5/51 (10%) 

Adjusted rate 0.0% 6.4% 13.5% 18.5% 

Terminal rate 0/22 (0%) 1/26 (4%) 5/37 (14%) 3/23 (13%) 

First incidence (days) — 645 740 (T) 720 

Logistic regression test P=0.042 P=0.241 P=0.095 P=0.036 

* Significantly different (P 0.05) from the control group by the Fisher exact test (interim evaluation) or the logistic regression test (2-year study) 

** P 0.01 

(T)Terminal sacrifice 

a Number of animals with lesion 

b Average severity of lesions in affected animals: 1 = minimal; 2 = mild; 3 = moderate; 4 = marked c Historical incidence for 2-year NTP feed studies with untreated controls (mean ± standard deviation): 30/1,301 (2.3% ± 2.9%); range, 0%-10% d Number of animals with neoplasm per number of animals with liver examined microscopically e Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality f Observed incidence in animals surviving until the end of the study 

g	 In the control column are the P values associated with the trend test. In the exposed group columns are the P values corresponding to the pairwise 

comparisons between the controls and that exposed group. The logistic regression test regards lesions in animals dying prior to terminal kill as 

nonfatal. 

h Not applicable; no neoplasms in animal group i Historical incidence: 9/1,301 (0.7% ± 1.5%); range, 0%-6% 
 

D&C Yellow No. 11, NTP TR 463 37 

Kidney: Two renal tubule adenomas in 5,000 pp m 

males and one renal tubule carcinoma in a 1,700 ppm 

male were observed in the standard (single section ) 

evaluation (Tables 6 and A1). Because of thi s 

suggestion of a chemical-related increase in rena l 

tubule neoplasms in males, an extended evaluatio n 

(step sections) of the kidney was conducted. Durin g 

the extended evaluation, two additional renal tubul e 

adenomas were observed in 5,000 ppm males, fou r 

renal tubule adenomas were observed in 1,700 pp m 

males, and two renal tubule adenomas were observed 

in 500 ppm males. No renal tubule neoplasms wer e 

observed in male controls. Renal tubule adenoma s 

(Plate 5) were more than five times the diameter of a 

normal tubule, usually had more complex structure s 

than hyperplasias, and often consisted of clusters o f 

multiple tubule-like structures. The one renal tubul e 

carcinoma in a 1,700 ppm male was approximatel y 

0.5 cm in diameter and was composed of atypica l 

epithelial cells forming solid clusters or abnorma l 

tubule-like structures that invaded the adjacent rena l 

parenchyma. One renal tubule carcinoma als o 

occurred in a 1,700 ppm female (Tables 6 and B1) . 

Renal tubule hyperplasia was observed in expose d 

groups of males but not in controls, and the incidences 

in 1,700 ppm males from both standard and extended 

evaluations were significantly greater than those in the 

controls (Table 6). Renal tubule hyperplasia was a 

discrete lesion ranging from a solid cluster of epithelial 

cells two to three times the di ameter of a normal tubule 

to a cystic lesion consisting of a tubule dilated up t o 

five times the normal diameter and lined with multiple 

layers of epithelial cells. 

At 12 months and 2 years, nephropathy was observed 

in all control and exposed male r ats and in most female 

rats (Tables 6, A5, and B5). The severity of nephropathy in exposed males and females was significantl y 

greater than in the controls, and the severity wa s 

greater in males than in females. Nephropath y 

included necrosis and regeneration of renal tubul e 

epithelium, typically with increased thickness o f 

basement membrane around regenerative tubules ; 

dilated tubules usually containing proteinaceous fluid; 

and interstitial fibrosis and inflammatory cel l 

aggregates. At 2 years, the incidences of hyperplasia 

of transitional epithelium of the kidney, whic h 

commonly accompanies advanced nephropathy, wer e 

greater in exposed males and 1,700 ppm females than 

in the controls, and the severity of this lesion i n 

exposed males and females was greater than in th e 

controls. Increased incidences of hyperplasia of th e 

parathyroid gland (0 ppm, 3/47; 500 ppm, 9/47 ; 

1,700 ppm, 15/48; 5,000 ppm, 17/52) and fibrou s 

osteodystrophy of the bone (2/50, 8/51, 18/51, 14/54) 

in exposed males at 2 years (Table A5) were probably 

secondary to the impaired kidney function associate d 

with nephropathy. 

The incidences of renal tubule pigmentation in al l 

exposed groups of males and females at 12 month s 

and 2 years were significantly greater than those in the 

controls (Tables 6, A5, and B5). Pigmentation of the 

renal tubule epithelium was yellow to brown granular 

material within the cytoplasm of cortical tubul e 

epithelial cells (Plate 6) and was similar in appearance 

to that seen in the liver. 
38 D&C Yellow No. 11, NTP TR 463 

TABLE 6 

Incidences of Neoplasms and Nonneoplastic Lesions of the Kidney in Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Male 

12-Month Interim Evaluation 

Number Examined Microscopically 

Nephropathya 1010 (1.1)b 

9

 9 (1.9)**

 9

 9 (2.6)**

 6 

6 (2.3)** 

Renal Tubule, Pigmentation 0 7** (1.0) 9** (1.8) 6** (2.5) 

2-Year Study 

Single Sections (Standard Evaluation) 

Number Examined Microscopically 50 51 51 54 

Nephropathy 50 (2.3) 51 (2.8)** 51 (3.2)** 54 (3.0)** 

Renal Tubule, Hyperplasia 0 0 4* (2.0) 3 (2.7) 

Renal Tubule, Pigmentation 18 (2.1) 43** (1.8) 47** (2.3) 54** (2.5) 

Transitional Epithelium, Hyperplasia 11 (1.3) 23** (1.8) 29** (1.9) 34** (1.7) 

Renal Tubule Adenomac 

Overall rated 0/50 (0%) 0/51 (0%) 0/51 (0%) 2/54 (4%) 

Renal Tubule Carcinoma 

Overall rate 0/50 (0%) 0/51 (0%) 1/51 (2%) 0/54 (0%) 

Step Sections (Extended Evaluation) 

Number Examined Microscopically 50 51 51 54 

Renal Tubule, Hyperplasia 0 2 (1.5) 9** (1.6) 2 (1.0) 

Renal Tubule Adenoma 

Overall rate 

Adjusted ratee 

Terminal ratef 

First incidence (days) 

Logistic regression testg 

0/50 (0%) 

0.0% 

0/19 (0%) 

—h 

P=0.259 

2/51 (4%) 

7.1% 

1/20 (5%) 

558 

P=0.255 

4/51 (8%) 

22.5% 

1/8 (13%) 

649 

P=0.046 

2/54 (4%) 

18.8% 

0/2 (0%) 

678 

P=0.120 

Single Sections and Step Sections (Combined) 

Number Examined Microscopically 50 51 51 54 

Renal Tubule, Hyperplasia 0 2 13** 4* 

Renal Tubule Adenoma 

Overall rate 0/50 (0%) 2/51 (4%) 4/51 (8%) 4/54 (7%) 

Adjusted rate 0.0% 7.1% 22.5% 38.3% 

Terminal rate 0/19 (0%) 1/20 (5%) 1/8 (13%) 0/2 (0%) 

First incidence (days) — 558 649 658 

Logistic regression test P=0.032 P=0.255 P=0.046 P=0.014 

Renal Tubule Carcinoma 

Overall rate 0/50 (0%) 0/51 (0%) 1/51 (2%) 0/54 (0%) 

Renal Tubule Adenoma or Carcinoma 

Overall rate 0/50 (0%) 2/51 (4%) 5/51 (10%) 4/54 (7%) 

Adjusted rate 0.0% 7.1% 26.1% 38.3% 

Terminal rate 0/19 (0%) 1/20 (5%) 1/8 (13%) 0/2 (0%) 

First incidence (days) — 558 649 658 

Logistic regression test P=0.036 P=0.255 P=0.022 P=0.014 

(continued) 
D&C Yellow No. 11, NTP TR 463 39 

TABLE 6 

Incidences of Neoplasms and Nonneoplastic Lesions of the Kidney in Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 (continued) 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Female 

12-Month Interim Evaluation 

Number Examined Microscopically 10 9 10 9 

Nephropathy 6 (1.0) 6 (1.0) 7 (1.1) 9 (1.2) 

Renal Tubule, Pigmentation 0 9** (1.7) 10** (2.2) 9** (3.0) 

Transitional Epithelium, Hyperplasia 0 0 0 2 (1.5) 

2-Year Study 

Number Examined Microscopically 50 51 50 51 

Nephropathy 45 (1.4) 47 (1.7)* 46 (1.8)** 50* (2.1)** 

Renal Tubule, Pigmentation 10 (1.3) 48** (1.8) 50** (2.8) 51** (3.2) 

Transitional Epithelium, Hyperplasia 2 (1.0) 6 (1.3) 10* (1.5) 3 (2.3) 

Renal Tubule Carcinomai 

Overall rate 0/50 (0%) 0/51 (0%) 1/50 (2%) 0/51 (0%) 

* Significantly different (P 0.05) from the control group by the Fisher exact test (incidences at interim evaluation), the logistic regression test 

(incidences at 2 years), or the Mann-Whitney U test (severity of nephropathy) 

** P 0.01 a Number of animals with lesion 

b Average severity of lesions in affected animals: 1 = minimal; 2 = mild; 3 = moderate; 4 = marked c Historical incidence for 2-year NTP feed studies with untreated controls (mean ± deviation): 9/1,301 (0.7% ± 1.5%); range, 0%-6% d Number of animals with neoplasm per number of animals with kidney examined microscopically e Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality f Observed incidence in animals surviving until the end of the study 

g	 In the control column are the P values associated with the trend test. In the exposed group columns are the P values corresponding to the pairwise 

comparisons between the controls and that exposed group. The logistic regression test regards lesions in animals dying prior to terminal kill as 

nonfatal. 

h Not applicable; no neoplasms in animal group i Historical incidence: 1/1,298 (0.1% ± 0.4%); range, 0%-2% 
40 D&C Yellow No. 11, NTP TR 463 

Oral Cavity (Oral Mucosa and Tongue): Squamous were observed in the oral cavity (oral mucosa o r 

cell carcinomas of the tongue were observed in on e tongue) in one control, one 500 ppm, two 1,700 ppm, 

500 ppm male at 12 months and one 5,000 pp m and four 5,000 ppm males; this lesion was als o 

female at 2 years (Tables 7, A1, and B1). One squa- observed in one control and one 500 ppm femal e 

mous cell carcinoma of the oral mucosa was observed (Tables 7 and B1). The incidence of squamous cel l 

in each group of exposed males and in one female i n papilloma or squamous cell carcinoma (combined) in 

the 5,000 ppm group at 2 years. Observations o f 1,700 and 5,000 ppm males exceeds the NT P 

squamous cell carcinoma of the oral cavity in mal e historical control range (0% to 4%, Table A4c) . 

F344/N rats are extraordinarily unusual because thi s Squamous cell papilloma was a discrete mass of thick, 

lesion has never been observed in 1,304 historica l branching epithelium overlying a central connectiv e 

control males from NTP feed studies (Table A4c) . tissue core with a stalk-like connection to the mucosal 

The incidence for 5,000 ppm fema le rats also exceeded surface (Plate 8). Hyperplasia was identified in th e 

the historical control range (Table B4c). Squamou s oral mucosa of one 5,000 ppm male at the 12-mont h 

cell carcinoma was an irregular mass composed o f interim evaluation and in the tongue of two 5,000 ppm 

thick cords and solid clusters of atypical epithelial cells males at 2 years. Hyperplasia was characterized by an 

that invaded the underlying connective tissue (Plate 7). increased number of cell layers of mucosal epithelium. 

At 2 years, squamous cell papillomas 

TABLE 7 

Incidences of Neoplasms and Nonneoplastic Lesions of the Oral Cavity in Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Male 

12-Month Interim Evaluation 

Oral Mucosaa 

Hyperplasiab 

0 

0 

0 

0 

0 

0 

1 

1 (2.0)c 

Tongue 0 1 0 0 

Squamous Cell Carcinoma 0 1 0 0 

2-Year Study 

Tongue 1 0 1 4 

Hyperplasia 0 0 0 2 (2.5) 

Oral Cavity (Oral Mucosa or Tongue) 

Squamous Cell Papilloma 

Overall rated 

Adjusted ratee 

Terminal ratef 

1/50 (2%) 

5.3% 

1/19 (5%) 

1/51 (2%) 

3.2% 

0/20 (0%) 

2/51 (4%) 

6.5% 

0/8 (0%) 

4/54 (7%) 

28.1% 

0/2 (0%) 

First incidence (days) 

Logistic regression testg 

733 (T) 

P=0.087 

658 

P=0.755 

406 

P=0.606 

481 

P=0.110 

Squamous Cell Carcinomah 

Overall rate 0/50 (0%) 1/51(2%) 1/51 (2%) 1/54 (2%) 

Squamous Cell Papilloma or Squamous Cell Carcinomai 

Overall rate 1/50 (2%) 2/51 (4%) 3/51 (6%) 5/54 (9%) 

Adjusted rate 5.3% 6.9% 10.6% 30.4% 

Terminal rate 1/19 (5%) 0/20 (0%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 733 (T) 658 406 481 

Logistic regression test P=0.066 P=0.487 P=0.369 P=0.069 

(continued) 
D&C Yellow No. 11, NTP TR 463 41 

TABLE 7 

Incidences of Neoplasms and Nonneoplastic Lesions of the Oral Cavity in Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 (continued) 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Female 

2-Year Study 

Oral Cavity (Oral Mucosa or Tongue) 

Squamous Cell Papilloma 

Overall rate 1/50 (2%) 1/51 (2%) 0/50 (0%) 0/51 (0%) 

Squamous Cell Carcinomaj 

Overall rate 0/50 (0%) 0/51 (0%) 0/50 (0%) 2/51 (4%) 

Squamous Cell Papilloma or Squamous Cell Carcinomak 

Overall rate 1/50 (2%) 

Adjusted rate 4.5% 

Terminal rate 1/22 (5%) 

First incidence (days) 740 (T) 

Logistic regression test P=0.332 

1/51 (2%) 

2.3% 

0/26 (0%) 

628 

P=0.757N 

0/50 (0%) 

0.0% 

0/37 (0%) 

)

l 

P=0.396N 

2/51 (4%) 

8.2% 

1/23 (4%) 

733 

P=0.518 

(T)Terminal sacrifice

a Number of animals with tissue examined microscopically b Number of animals with lesion c Average severity of lesions in affected animals: 1 = minimal; 2 = mild; 3 = moderate; 4 = marked d Number of animals with neoplasm per number of animals necropsied e Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality f Observed incidence in animals surviving until the end of the study 

g	 In the control column are the P values associated with the trend test. In the exposed group columns are the P values corresponding to the pairwise 

comparison between the controls and that exposed group. The logistic regression test regards lesions in animals dying prior to terminal kill as 

nonfatal. A lower incidence in an exposure group is indicated by N. h Historical incidence for 2-year NTP feed studies with untreated control groups: 0/1,304 i Historical incidence (mean ± standard deviation): 10/1,304 (0.8% ± 1.3%); range, 0%-4% j Historical incidence: 4/1,301 (0.3% ± 0.7%); range, 0%-2% k Historical incidence: 15/1,301 (1.2% ± 1.6%); range, 0%-6% l Not applicable; no neoplasms in animal group 

Testis: There was an exposure-related increase in the forestomach varied in severity and extent, rangin g 

incidences of testicular adenoma at 2 years, and th e from minimal focal lesions at the limiting ridge of the 

incidence of this lesion in each exposed group wa s mucosa to marked lesions affecting cell layer s 

significantly greater than that in the controls (0 ppm , throughout the forestomach mucosa. In the smal l 

39/49; 500 ppm, 46/51; 1,700, 48/51; 5,000 ppm , intestine of 1,700 and 5,000 ppm males at 2 years, the 

46/54; Table A3). Because the incidences within the incidences of epithelial hyperplasia of the duodenu m 

historical control range (74% to 98%; Table A4d) are (1/50, 4/51, 22/51, 21/54), jejunum (0/50, 3/50 , 

so high, the significance of the incidences in th e 10/51, 12/54), and ileum (0/49, 3/50, 10/50, 8/54 ) 

exposed groups is unclear. were greater than those in the controls. Hyperplasia of 

the small intestine consisted o f a diffuse increase in the 

Forestomach and Small Intestine: The incidences of number of villous epithelial cells, which appeare d 

mucosal hyperplasia of the forestomach in expose d crowded together and taller, and an increase in th e 

males (4/50, 13/51, 19/51, 21/54; Table A5) an d height of the villous projections. This lesion wa s 

females (5/50, 17/51, 30/50, 27/51; Table B5) wer e usually apparent on gross examination because of the 

greater than those in the controls at 2 years . greater diameter and thicker mucosa of affecte d 

Hyperplasia of the squamous epithelium of th e intestines. 
42 D&C Yellow No. 11, NTP TR 463 

Salivary Gland and Pancreas: Incidences of atrophy of 

the salivary glands were greater than those in th e 

controls in 5,000 ppm males (1/50, 1/51, 5/50, 7/54; 

Table A5) and in all exposed groups of females (0/50, 

8/46, 7/50, 13/50; Table B5). Atrophy was a minimal 

to mild focal to multifocal decrease in the size o f 

glandular acini accompanied by increased amounts of 

interstitial connective tissue between acini. In th e 

pancreas, incidences of cytoplasmic alteration of th e 

acinar cell in exposed males were significantly greater 

than that in the controls (0/50, 5/51, 11/51, 8/54 ; 

Table A5). Cytoplasmic alteration of the acinar cells 

of the pancreas was a diffuse loss of zymogen granules 

from the cytoplasm. This change might reflect debilitation because all males in which this change occurred 

died before the end of the study. The incidences (9/50, 

19/51, 17/50, 14/51; Table B5), but not the severity, 

of pancreatic atrophy were greater in exposed females 

than in controls. Pancreatic atrophy was characterized 

by a decrease in the size of pancreatic acini and a 

relative increase in the amount of connective tissu e 

between acini. 

Lymph Nodes: At 2 years, the incidences of lymphoid 

hyperplasia were greater than those in the controls i n 

the mandibular lymph nodes in 1,700 and 5,000 ppm 

males (8/50, 12/51, 22/50, 25/53) and females (5/50, 

10/51, 14/50, 13/49), the mesenteric lymph nodes i n 

1,700 and 5,000 ppm males (3/50, 2/50, 10/51 , 

14/54), and the mediastinal lymph nodes in 1,700 and 

5,000 ppm males (0/20, 0/23, 9/26 , 17/45) (Tables A5 

and B5). Mediastinal and pancreatic lymph node s 

were examined microscopically only when they wer e 

grossly abnormal. Lymphoid hyperplasia was de ­

scribed as an increase in the size (1.5 to 2 time s 

normal) of lymph nodes, which was usually accompanied by an increase in the density of cortical lymphocytes. This lymphoid hyperplasia suggests an immune 

response was associated with the administration o f 

D&C Yellow No. 11 in some individuals. In 1,70 0 

and 5,000 ppm males, the incidences of hemorrhage of 

the mesenteric lymph nodes (0/50, 0/50, 9/51, 7/54 ) 

and mediastinal lymph nodes (0/20, 3/23, 7/26, 16/45) 

were greater than those in the controls. Hemorrhag e 

within lymph nodes consisted of small to moderat e 

numbers of extravascular red blood cells withi n 

medullary sinuses. Incidences of pigmentation of the 

pancreatic lymph nodes in 5,000 ppm males (1/20 , 

3/23, 3/26, 12/45) and females (1/9, 2/11, 6/11, 8/15) 

were greater than in the controls. Pigmentation within 

lymph nodes was described as yellow to brown granular material within the cytoplasm of macrophages. 

Clitoral Gland: At 2 years, the incidences of clitora l 

gland adenoma (11/49, 4/50, 5/49, 4/51) and clitora l 

gland adenoma or carcinoma (combined) (17/49, 6/50, 

11/49, 6/51) in exposed groups of females wer e 

significantly less than those in the controls (Table B3). 

There was a negative trend in the incidences of ade ­

noma or carcinoma (combined); howeve r, the incidence 

in controls exceeded the previous historical contro l 

range (2% to 21%; Table B4d). The significance o f 

this finding is uncertain. 

Pituitary Gland: The incidences of pars distali s 

adenoma in 500 and 5,000 ppm males (20/50, 8/50 , 

14/50, 10/52) were significantly less than that in th e 

controls; the trend was not significant (Table A3). 

All Organs: At 2 years, the incidences of mononuclear 

cell leukemia in 1,700 and 5,000 ppm males wer e 

significantly less than that in the controls by th e 

logistic regression test (37/50, 36/51, 20/51, 22/54 ; 

Table A3). However, the decreased incidences wer e 

not significant by the life table test (the most appro ­

priate test for this generally fatal neoplasm) and were 

considered to be due primarily to reduced survival i n 

these groups. Similar effects were not observed i n 

female rats (Table B3). 

GENETIC TOXICOLOGY 

Results of mutagenicity tests with D &C Yellow No. 11 

in Salmonella typhimurium were equivocal in one 

study, based on the responses observed in strai n 

TA100 with 10% induced rat liver S9, and the results 

were weakly positive in a second study, which use d 

slightly lower doses, based on responses observed i n 

strains TA98 and TA100 with 30% induced rat o r 

hamster liver S9 (Table C1; Zeiger et al., 1988). No 

indication of mutagenic activity was observed in th e 

absence of S9 in any of the strains tested. The dat a 

from the S. typhimurium studies indicate variable 

responses among replicate trials within a particula r 

treatment condition; this may have been the result o f 

precipitate formation at higher concentrations (333 μg/ 
D&C Yellow No. 11, NTP TR 463 43 

plate and above) and consequent variability in th e with S9), the frequency of aberrant cells, and th e 

actual D&C Yellow No. 11 exposure concentrations. difficulty in finding scorable cells in some cases (Trial 

1, without S9). 

In cytogenic tests with cultured Chinese hamster ovary 

cells, D&C Yellow No. 11 induced highly significant Despite the strong response seen in the in vitro chroincreases in both sister chromatid exchange s mosomal aberrations assay, no increase in the fre ­

(Table C2) and chromosomal aberrations (Table C3 ) quency of micronucleated normochromatic erythro ­

with and without S9. Cell cycle delay, requiring a n cytes was observed in peripheral blood samples from 

extended incubation period, was observed in the sister male and female mice given D&C Yellow No. 11 i n 

chromatid exchange test at doses of 1.5 μg/mL an d feed for 13 weeks (Table C4). 

above; in the chromosomal aberrations test, no dela y 

was observed in the absence of S9, but cultures treated In conclusion, D&C Yellow No. 11 was mutagenic in 

in the presence of S9 were harvested late because cell bacteria and clastogenic in mammalian cells in vitro, 

cycle delay was anticipated. Less than 200 cells pe r but no evidence of clastogenicity was observed in the 

dose level were scored in all but one dose level in the single in vivo study performed with male and femal e 

chromosomal aberrations test due to the high number mice. 

of chromosomal aberrations per cell (cultures treate d 
 

 

 

 

 

 

PLATE 1 

Hepatocellular adenoma (arrow) in a female F344/N rat exposed to 500 ppm D&C 

Yellow No. 11 in feed for 2 years. Note the altered architecture and abruptly 

intersected and compressed normal hepatic cords (normal to left). H&E; 100× 

PLATE 3 

Clear cell focus in the liver of a female F344/N rat exposed to 5,000 ppm D&C 

Yellow No. 11 in feed for 2 years. Note the hepatocytes within the roughly circular 

focus (arrows) are slightly larger and have relatively clear cytoplasm and central 

nuclei. H&E; 120× 

PLATE 2 

Edge of a hepatocellular carcinoma (arrow) in a female F344/N rat exposed to 

5,000 ppm D&C Yellow No. 11 in feed for 2 years. Note that neoplastic hepatocytes 

are in islands and clusters rather than cords. H&E; 100× 

PLATE 4 

Pigmentation (dark granular material) in hepatocytes (arrows) and Kupffer’s cells in 

a male F344/N rat exposed to 5,000 ppm D&C Yellow No. 11 in feed for 2 years.

 H&E; 240× 
 

 

 

 

 

 

PLATE 5 

Renal tubule adenoma in a male F344/N rat exposed to 1,700 ppm D&C Yellow 

No. 11 in feed for 2 years. Note the solid clusters of neoplastic epithelial cells in the 

connective tissue of a markedly nephrotic kidney. H&E; 100× 

PLATE 7
 

Squamous cell carcinoma of the oral cavity in a male F344/N rat exposed to 5,000 

ppm D&C Yellow No. 11 in feed for 2 years. Note that the nests of neoplastic 

squamous cells (arrows) invaded the underlying connective tissue. H&E; 30× 

PLATE 6 

Pigmentation in the kidney of a male F344/N rat exposed to 5,000 ppm D&C Yellow 

No. 11 in feed for 2 years. Note the granular dense material that stained brown in the 

cytoplasm of renal tubule epithelial cells (arrows). H&E; 240× 

PLATE 8
 

Squamous cell papilloma of the oral mucosa in a female F344/N rat exposed to 500
 

ppm D&C Yellow No. 11 in feed for 2 years. Note that the super-ficially keratinized
 

squamous epithelium proliferates on a branching fibrous connective tissue stalk.
 

H&E; 17×
 
44 D&C Yellow No. 11, NTP TR 463 
45 

DISCUSSION AND CONCLUSIONS
 

D&C Yellow No. 11 was nominated for dosed-fee d 

toxicity and carcinogenicity studies because it is a 

contaminant in D&C Yellow No. 10, a widely used , 

high-production dye which could potentially be in ­

gested. Ingestion of D&C Yellow No. 11 ha s 

produced effects on growth in rats and mice. In th e 

parental generation (F ) in the current studies, mea n 0 

body weight gains of exposed males and 5,000 pp m 

females were less than those of controls prior t o 

cohabitation, but feed consumption by exposed rat s 

was generally similar to that by the controls. Ther e 

were no apparent D&C Yellow No. 11 effects o n 

survival, reproductive performance (i.e., duration o f 

gestation, average litter size, number of live pups per 

litter on days 4 or 21, or percentage of male pups), or 

mean litter weights at 1 or 4 days of age. However, by 

14 days of age, the mean litter weights of all exposed 

groups were less than those of the controls. Although 

feed consumption by exposed and controls groups was 

similar, mean body weights of 1,700 and 5,000 pp m 

males and females were generally lower than those of 

controls throughout the 2-year study. A simila r 

response was observed in the previous perinata l 

exposure study in F344/N rats (NTP, 1991a). In that 

study, the dams were given 0, 5,000, 17,000 or 50,000 

ppm D&C Yellow No. 11 in feed for 4 weeks befor e 

mating (sires were not exposed) and throughou t 

gestation, lactation, and weaning (day 28 afte r 

birthing). Pups were continued at the same exposure 

concentrations as their dams for 4 weeks afte r 

weaning. Mean pup body weights were similar a t 

birth, but group mean body weights were less tha n 

those of the controls in all exposed groups at 4 weeks 

of age and in the 17,000 and 50,000 ppm groups at 8 

weeks of age. However, in 13-week feed studie s 

without perinatal exposure in F3 44/N rats and B6C3F 1 

mice, up to 50,000 ppm D&C Yellow No. 11 in feed 

did not affect mean body weight gains or fee d 

consumption (NTP, 1991a). The decreased bod y 

weight gains in rats are most likely a chemical-related 

phenomenon that affects optim um feed utilization (i.e., 

absorption and metabolism) rather than the result o f 

decreased feed consumption. The effect of D& C 

Yellow No. 11 on body weight gains seems to be more 

pronounced when rats are exposed perinatally or i n 

feed at a very young age. 

After oral administration of radiolabeled D&C Yellow 

No. 11 to F344/N rats, the dye was rapidly absorbe d 

and distributed to all tissues, and 98% was excrete d 

within 72 hours (El Dareer et al., 1988). D&C Yellow 

No. 11 was excreted in the feces and urine, and th e 

liver and kidney had greater concentrations o f 

radioactivity than did other tissues. More than 50% of 

an intravenous dose of D&C Yellow No. 11 wa s 

excreted via the bile 4 hours after administratio n 

(El Dareer et al., 1988). No parent compound was recovered, and more than 10 metabolites were identified 

in the bile. The target organs in the current 2-yea r 

study are associated with known pathways of ingested 

D&C Yellow No. 11 [i.e., oral cavity (portal of entry), 

liver (metabolism), and bile duct and kidne y 

(excretion)]. These organ sites were also the primary 

targets identified in short-term oral toxicity studies . 

Pigment was shown to accumulate in the liver, bil e 

duct, and kidney of 8-week-old rats perinatally ex ­

posed followed by exposure to D&C Yellow No. 11 in 

feed after weaning and in dosed-feed studies in rat s 

and mice with no perinatal exposure (NTP, 1991a ; 

Eastin et al., 1996). In addition, liver and kidne y 

weights were increased in rats and mice after ora l 

exposure to D&C Yellow No. 11, which suggest s 

elevated metabolic and excretory activities in thes e 

organs. 

Periportal degeneration of hepatocytes was apparent in 

rats examined at 8 weeks of age in the previou s 

perinatal exposure study, in rats in the 14-day an d 

13-week studies (NTP, 1991a), and at the 12-mont h 

interim evaluation in the current study. Hepatocellular 

cytologic alteration (cytoplasmic basophilia an d 

granularity) and pigmentation in less affected livers in 

the chronic studies also exhibited a periporta l 

distribution. These alterations are clear indications of 

liver toxicity, but widespread hepatocellular necrosi s 

was not seen at any time point studied. It was no t 

possible to determine a zonal distribution of hepato ­

cellular foci or neoplasms in the 2-year study. Th e 
46 D&C Yellow No. 11, NTP TR 463 

incidences of liver adenomas in 5,000 ppm males and male rats. Other characteristic features of hydrocarbon 

females exceeded the historical control ranges. nephropathy, including regeneration/necrosis, granular 

casts, and homogenous protein casts in renal tubules, 

The marginal indication of a neoplastic effect, coinci- were not observed in the current study. It appears that 

dent with exposure-related exacerbated severity o f not all chemicals causing an increase in hyaline droplet 

nephropathy and increased pigmentation of the rena l accumulation may have the same spectrum of rena l 

tubule, in the standard evaluation (single sections ) toxicity described for hydrocarbon nephropathy . 

prompted an extended evaluation (step sections) of the Exposure to p-nitrobenzoic acid in feed for 13 weeks 

kidney in male rats. The combined results of th e (NTP, 1994) also caused hyaline droplet accumulation 

standard and extended evaluations indicated a modest in male rats; however, granular casts, necrosis, an d 

chemical-related increase in renal tubule neoplasms . regeneration were not evident, and there was n o 

The yellow-brown pigment was most likely D& C chemical-related exacerbation of nephropathy durin g 

Yellow No. 11 or a metabolite because special stain s the 2-year study. 

of similar pigmentation in the 13-week studies were all 

negative for bile, hemosiderin, and lipofuscin. Cyto - D&C Yellow No. 11 has been shown to have ski n 

plasmic alteration (an increase in size and number o f sensitization and allergenic potential (Lamson et al., 

cytoplasmic hyaline droplets) was also present in th e 1982; Kita et al., 1984). The finding of an association 

renal tubule epithelium in all exposed male groups . between D&C Yellow No. 11 exposure and oral cavity 

These hyaline droplets often formed large globules or neoplasms in the current study was unexpected . 

irregularly shaped crystalline structures that staine d However, it is possible that prolonged contact with a 

similarly (Mallory-Heidenhain method) to the smaller chemical shown to be a slight skin irritant and whic h 

granules of protein ( 2μ-globulin) typically seen in the has mutagenic and clastogenic activity with and withrenal tubule cell cytoplasm of male F344/N rats. In a out metabolic activation could have produced th e 

separate study (Eastin et al., 1996), male rats given response observed in the oral cavity. The numbers of 

5,000 ppm D&C Yellow No. 11 in feed for 70 day s papillomas of the tongue and oral mucosa in male rats 

had cytoplasmic alteration and pigment in the rena l are small, but the presence of an oral cavity carcinoma 

tubules and hepatocellular degeneration and pigmenta- in one 500 ppm male at 12 months, in each of th e 

tion similar to that seen at the same exposure concen- exposed male groups at 2 years, plus carcinomas i n 

tration in the 13-week study. After a recovery period two 5,000 ppm female rats at 2 years, as well as th e 

during which rats were given undo sed feed for up to 28 low rates of these neoplasms in historical control s 

days, pigment was still present in the liver biliar y suggest that exposure to D&C Yellow No. 11 in feed 

epithelium and renal tubule epithelium, and cytoplas- is associated with neoplastic proliferation of th e 

mic alteration and pigment in the renal tubule epithe- epithelium in the oral cavity. The incidence of squa ­

lium were reduced in severity in all rats. There was no mous cell carcinoma in the oral cavity of 5,000 pp m 

immunohistochemical evidence of an increase in th e females (2/51) exceeds the NTP historical control rate 

amount of 2μ-globulin in the kidney of rats wit h (4/1,301) and suggests that the neoplastic effec t 

cytoplasmic alteration after 28 days on undosed feed; occurred in males and females. 

cytoplasmic hyaline droplets were similar in th e 

controls. The cytoplasmic alteration, characterized by The cause of the edema of the head and neck, whic h 

increased size and number of irregularly shape d was observed grossly and resulted in the deaths of one 

hyaline droplets in the renal tubule epithelium of male male in the 1,700 ppm group and five males in th e 

F344/N rats, was morphologically similar to th e 5,000 ppm group, could not be determined. Possible 

abnormal accumulation of irregularly shaped hyalin e causes of edema could include local obstruction o f 

droplets containing 2μ-globulin that has been de - lymphatics or blood vasculature; heart failure; hypo ­

scribed as a feature of chlorinated hydrocarbo n proteinemia secondary to kidney or liver disease o r 

(“hyaline droplet”) nephropathy in male rats (NTP , intestinal malabsorption; or altered vascular pathways 

1991b; 1991c). Typically, chronic administration o f secondary to abnormal flow through the liver or t o 

chemicals causing this type of renal toxicity results in primary vasculopathy. Tissues from rats that had the 

enhanced severity of nephropathy and an increase i n diagnosis of edema were given complete histopatho ­

proliferative lesions of the renal tubule epithelium i n logic reviews without showing evidence of unusuall y 
__________ 

D&C Yellow No. 11, NTP TR 463 47 

severe heart, liver, or kidney disease, or of vasculop - incidences of hepatocellular neoplasms. Incidences of 

athy or consistent intestinal mucosal hyperplasia . uncommon squamous cell carcinoma of the oral cavity 

Since functional disturbances of fluid dynamics, heart in females may have been related to chemica l 

function, or vascular tone do not necessarily hav e treatment. 

histopathologic correlates, the edema may have been a 

physiologic effect of D&C Yellow No. 11 or it s 

metabolites.	 Exposure of rats to D&C Yellow No. 11 in feed for 2 

years resulted in increased incidences of nonneoplastic 

liver lesions including clear cell foci, increased baso -

CONCLUSIONS philia and granularity in the cytoplasm of hepatocytes, 

Under the conditions of this perinatal exposure fol - and bile duct, hepatocyte, and Kupffer cell pigmentalowed by a 2-year dosed feed study, there was some tion in males and females and mixed cell foci in males. 

evidence of carcinogenic activity * of D&C Yellow In the kidney, there were increased incidences of renal 

No. 11 in male F344/N rats based on increased inci - tubule pigmentation and transitional epithelial hyperdences of hepatocellular adenoma, renal tubule neo - plasia in males and females and renal tubule hyperplaplasms, and squamous cell neoplasms of the ora l sia in males. The severity of nephropathy was in ­

cavity. There was some evidence of carcinogeni c creased in exposed males and females. 

activity in female F344/N rats based on increase d 

*	 Explanation of Levels of Evidence of Carcinogenic Activity is on page 9. A summary of the Technical Reports Review Subcommittee comments and 

the public discussion on this Technical Report appears on page 11. 
48 D&C Yellow No. 11, NTP TR 463 
49 

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and Methods for their Dete ction (A. Hollaender, Ed.), 

Vol. 4, pp. 31-53. Plenum Press, New York. 

Shirley, E. (1977). A non-parametric equivalent o f 

Williams’ test for contrasting increasing dose levels of 

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Straus, D.S. (1981). Somatic mutation, cellular differentiation, and cancer causation. JNCI 67, 233-241. 
52 D&C Yellow No. 11, NTP TR 463 

Sun, J.D., Henderson, R.F., Marshall, T.C. , 

Cheng, Y.-S., Dutcher, J.S., Pickrell, J.A. , 

Mauderly, J.L., Hahn, F.F., Banas, D.A., Seiler, F.A., 

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Tarone, R.E. (1975). Tests for trend in life tabl e 

analysis. Biometrika 62, 679-682. 

Tennant, R.W., Margolin, B.H., Shelby, M.D., Zeiger, 

E., Haseman, J.K., Spalding, J., Caspary, W., Resnick, 

M., Stasiewicz, S., Anderson, B., and Minor, R . 

(1987). Prediction of chemical carcinogenicity i n 

rodents from in vitro genetic toxicity assays. Science 

236, 933-941. 

Weaver, J.E. (1983). Dose response relationships i n 

delayed hypersensitivity to quinoline dyes. Contact 

Dermatitis 9, 309-312. 

Williams, D.A. (1971). A test for differences between 

treatment means when several dose levels ar e 

compared with a zero dose control. Biometrics 27, 

103-117. 

Williams, D.A. (1972). The comparison of severa l 

dose levels with a zero dose control. Biometrics 28, 

519-531. 

Yuan, J., Jameson, C.W., Goehl, T.J., Elwell, M.R. , 

Leininger, J.R., Thompson, M.B., Corniffe, G., an d 

Carlton, T. (1992). Application of molecular encap ­

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of p-chloro­ , , -trifluorotoluene in -cyclodextrin 

vehicle versus corn oil vehicle in male and femal e 

Fischer 344 rats and B6C3F1 mice. Fundam. Appl. 

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Mortelmans, K. (1988). Salmonella mutagenicity 

tests: IV. Results from the testing of 300 chemicals . 

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Zeiger, E., Haseman, J.K., Shelby, M.D. , 

Margolin, B.H., and Tennant, R.W. (1990). Evalua ­

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16 (Suppl. 18), 1-14. 
53 

APPENDIX A 

SUMMARY OF LESIONS IN MALE RATS 

IN THE 2-YEAR FEED STUDY 

OF D&C YELLOW NO. 11 

TABLE A1 

TABLE A2 

TABLE A3 

TABLE A4a 

TABLE A4b 

TABLE A4c 

TABLE A4d 

Summary of the Incidence of Neoplasms in Male Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

Individual Animal Tumor Pathology of Male Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

Statistical Analysis of Primary Neoplasms in Male Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

Historical Incidence of Hepatocellular Neoplasms 

in Untreated Male F344/N Rats .................................................... 

Historical Incidence of Renal Tubule Neoplasms 

in Untreated Male F344/N Rats .................................................... 

Historical Incidence of Oral Cavity Neoplasms 

in Untreated Male F344/N Rats .................................................... 

Historical Incidence of Testicular Adenoma 

55 

60 

80 

88 

88 

89 

TABLE A5 

in Untreated Male F344/N Rats .................................................... 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

89 

90 
54 D&C Yellow No. 11, NTP TR 463 
 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 55 

TABLE A1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 a 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Disposition Summary 

Animals initially in study 

12-Month interim evaluation 

Early deaths 

Moribund 

Natural deaths 

Other 

Survivors 

Terminal sacrifice 

60 

10 

29 

2 

19 

60 

9 

29 

2 

20 

60 

9 

41 

1 

1 

8 

60 

6 

49 

3 

2 

Animals examined microscopically 60 60 60 60 

12-Month Interim Evaluation 

Alimentary System 

Pancreas 

Acinar cell, adenoma

Tongue

Squamous cell carcinoma

(10) (9)

 (1) 

 1 (100%) 

(9) (6) 

 1 (17%) 

Endocrine System 

Pituitary gland 

Pars distalis, adenoma

(10)

 1 (10%) 

(9) (8) (6) 

Genital System 

Testes 

Interstitial cell, adenoma

(10) (9)

 3 (33%) 

(9)

 1 (11%) 

(6) 

Systems Examined With No Neoplasms Observed 

Cardiovascular System 

General Body System 

Hematopoietic System 

Integumentary System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

2-Year Study 

Alimentary System 

Intestine large, colon (50) (50) (50) (54) 

Polyp adenomatous 1 (2%) 

Intestine large, cecum (50) (50) (50) (54) 

Hemangioma 1 (2%) 

Intestine small, jejunum (50) (50) (51) (54) 

Carcinoma 1 (2%) 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

56 D&C Yellow No. 11, NTP TR 463 

TABLE A1
 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Alimentary System (continued) 

Intestine small, ileum (49) (50) (50) (54) 

Carcinoma 1 (2%) 

Liver (50) (51) (51) (54) 

Hepatocellular adenoma 1 (2%) 2 (4%) 1 (2%) 6 (11%) 

Hepatocellular adenoma, multiple 1 (2%) 

Mesentery (11) (12) (9) (13) 

Lipoma 1 (9%) 

Oral mucosa (2) (3) (3) 

Squamous cell carcinoma 1 (50%) 1 (33%) 1 (33%) 

Squamous cell papilloma 1 (50%) 1 (33%) 2 (67%) 

Pancreas (50) (51) (51) (54) 

Acinar cell, adenoma 1 (2%) 4 (8%) 1 (2%) 

Salivary glands (50) (51) (50) (54) 

Carcinoma, metastatic, Zymbal’s gland 1 (2%) 

Stomach, forestomach (50) (51) (51) (54) 

Squamous cell papilloma 2 (4%) 

Stomach, glandular (50) (51) (51) (54) 

Carcinoma 1 (2%) 

Tongue (1) (1) (4) 

Squamous cell papilloma 1 (100%) 1 (100%) 2 (50%) 

Cardiovascular System 

Heart (50) (51) (51) (54) 

Schwannoma malignant 1 (2%) 

Endocrine System 

Adrenal cortex (50) (51) (51) (54) 

Adrenal medulla (50) (50) (51) (54) 

Neuroblastoma malignant 1 (2%) 

Pheochromocytoma malignant 4 (8%) 1 (2%) 2 (4%) 

Pheochromocytoma benign 7 (14%) 8 (16%) 11 (22%) 6 (11%) 

Bilateral, pheochromocytoma benign 1 (2%) 2 (4%) 

Islets, pancreatic (50) (51) (51) (54) 

Adenoma 4 (8%) 3 (6%) 

Carcinoma 1 (2%) 2 (4%) 

Pituitary gland (50) (50) (50) (52) 

Pars distalis, adenoma 20 (40%) 7 (14%) 13 (26%) 8 (15%) 

Pars distalis, adenoma, multiple 1 (2%) 1 (2%) 2 (4%) 

Pars intermedia, adenoma 1 (2%) 1 (2%) 1 (2%) 

Thyroid gland (50) (51) (50) (54) 

C-cell, adenoma 5 (10%) 2 (4%) 3 (6%) 3 (6%) 

C-cell, adenoma, multiple 1 (2%) 1 (2%) 

C-cell, carcinoma 1 (2%) 1 (2%) 

Follicular cell, adenoma 1 (2%) 

Follicular cell, carcinoma 3 (6%) 1 (2%) 1 (2%) 3 (6%) 

General Body System 

Peritoneum (2) (1) (3) (3) 
 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 57 

TABLE A1
 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Genital System 

Epididymis (49) (51) (51) (54) 

Preputial gland (49) (50) (51) (54) 

Adenoma 5 (10%) 2 (4%) 1 (2%) 

Carcinoma 5 (10%) 4 (8%) 1 (2%) 2 (4%) 

Prostate (50) (51) (51) (53) 

Adenocarcinoma 1 (2%) 

Seminal vesicle (50) (51) (51) (54) 

Testes (49) (51) (51) (54) 

Bilateral, interstitial cell, adenoma 30 (61%) 36 (71%) 42 (82%) 39 (72%) 

Interstitial cell, adenoma 9 (18%) 10 (20%) 6 (12%) 7 (13%) 

Hematopoietic System 

Bone marrow (50) (51) (51) (54) 

Lymph node (20) (23) (26) (45) 

Renal, pheochromocytoma malignant,

 metastatic, adrenal medulla 1 (5%) 

Lymph node, mandibular (50) (51) (50) (53) 

Carcinoma, metastatic, Zymbal’s gland 1 (2%) 

Lymph node, mesenteric (50) (50) (51) (54) 

Spleen (50) (51) (51) (54) 

Hemangiosarcoma 1 (2%) 

Thymus (48) (49) (49) (53) 

Integumentary System 

Mammary gland (48) (48) (49) (53) 

Fibroadenoma 3 (6%) 2 (4%) 3 (6%) 1 (2%) 

Skin (50) (50) (51) (54) 

Basal cell adenoma 1 (2%) 1 (2%) 2 (4%) 

Basal cell carcinoma 2 (4%) 

Keratoacanthoma 7 (14%) 4 (8%) 2 (4%) 1 (2%) 

Squamous cell papilloma 3 (6%) 3 (6%) 2 (4%) 3 (6%) 

Trichoepithelioma 1 (2%) 

Sebaceous gland, adenoma 1 (2%) 

Sebaceous gland, carcinoma 1 (2%) 1 (2%) 

Subcutaneous tissue, fibroma 3 (6%) 2 (4%) 3 (6%) 

Subcutaneous tissue, lipoma 1 (2%) 

Subcutaneous tissue, schwannoma malignant 1 (2%) 

Musculoskeletal System 

Bone (50) (51) (51) (54) 

Osteosarcoma 1 (2%) 

Skeletal muscle (1) (2) (5) (9) 

Nervous System 

Brain (50) (51) (51) (54) 

Hemangioma 1 (2%) 

Spinal cord (1) (2) (1) 
 

 

 

 

 

 

 

58 D&C Yellow No. 11, NTP TR 463 

TABLE A1
 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar adenoma, multiple

Alveolar/bronchiolar carcinoma

Carcinoma, metastatic, Zymbal’s gland

Osteosarcoma, metastatic, bone

(50) 

 7 (14%) 

 1 (2%) 

 1 (2%) 

(51) 

 1 (2%) 

 1 (2%) 

 1 (2%) 

(51) 

 2 (4%) 

 1 (2%) 

(54) 

 1 (2%) 

 1 (2%) 

Special Senses System 

Zymbal’s gland

Carcinoma

 (1)

 1 (100%)

 (1) 

 1 (100%) 

Urinary System 

Kidney 

Osteosarcoma, metastatic, bone

Renal tubule, adenoma

Renal tubule, carcinoma

Urinary bladder 

(50) 

 1 (2%) 

(50) 

(51) 

(51) 

(51) 

 1 (2%) 

(51) 

(54) 

 2 (4%) 

(54) 

Systemic Lesions 

Multiple organsb 

Leukemia mononuclear

Lymphoma malignant

Mesothelioma malignant

(50) 

 37 (74%) 

 1 (2%) 

 2 (4%) 

(51) 

 36 (71%) 

 2 (4%) 

(51) 

 20 (39%) 

 3 (6%) 

(54) 

 22 (41%) 

 4 (7%) 
59 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE A1
 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Neoplasm Summary 

Total animals with primary neoplasmsc 

12-Month interim evaluation 1 4 1 1 

2-Year study 50 51 50 49 

Total primary neoplasms 

12-Month interim evaluation 1 4 1 1 

2-Year study 170 140 139 127 

Total animals with benign neoplasms 

12-Month interim evaluation 1 3 1 1 

2-Year study 49 50 49 49 

Total benign neoplasms 

12-Month interim evaluation 1 3 1 1 

2-Year study 111 92 102 89 

Total animals with malignant neoplasms 

12-Month interim evaluation 1 

2-Year study 42 39 30 26 

Total malignant neoplasms 

12-Month interim evaluation 1 

2-Year study 59 48 37 38 

Total animals with metastatic neoplasms 

2-Year study 2 1 

Total metastatic neoplasms 

2-Year study 3 3 

a Number of animals examined microscopically at the site and the number of animals with neoplasm b Number of animals with any tissue examined microscopically
 

Primary neoplasms: all neoplasms except metastatic neoplasms
 
 

 

 

 

 

 

 

 

 

 

 

60 D&C Yellow No. 11, NTP TR 463 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm 

4 5 5 5 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 7 

Number of Days on Study 8 0 3 9 0 1 1 2 2 2 2 3 3 3 3 5 5 5 6 7 8 9 9 9 0 

1 9 0 0 0 5 9 1 2 3 8 2 2 7 8 0 6 6 7 7 8 3 5 5 1 

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

Carcass ID Number 5 1 0 5 4 5 2 3 4 4 2 0 3 1 4 1 3 5 0 1 3 1 0 3 0 

3 1 1 4 4 8 8 9 5 0 9 4 4 5 7 8 2 0 5 6 6 7 7 7 8 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + M + + + + + + + + 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Intestine small, ileum + M + + + + + + + + + + + + + + + + + + + + + + + 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Hepatocellular adenoma 

Mesentery + + + + + 

Lipoma 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Tongue 

Squamous cell papilloma 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + +
 

Heart + + + + + + + + + + + + + + + + + + + + + + + + +
 

Schwannoma malignant X
 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 

Pheochromocytoma malignant X 

Pheochromocytoma benign X X X 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma 

Carcinoma 

Parathyroid gland + M + + + + + + + + + + + + + + + + + + + + + + M 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X X X X X X X X X 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

C-cell, adenoma X X 

C-cell, adenoma, multiple X 

Follicular cell, carcinoma X 

General Body System 

Peritoneum + + 

Genital System 

Epididymis + + + + + + M + + + + + + + + + + + + + + + + + + 

Preputial gland + + + + + + M + + + + + + + + + + + + + + + + + + 

Adenoma X X 

Carcinoma X 

+: Tissue examined microscopically M: Missing tissue X: Lesion present 

A: Autolysis precludes examination I: Insufficient tissue Blank: Not examined 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 61 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 0 0 0 1 1 2 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 

2 6 6 6 9 2 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 4 4 

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Total 

Carcass ID Number 4 0 2 3 2 5 0 0 1 1 1 1 2 2 2 2 3 3 3 4 4 5 5 5 6 Tissues/ 

3 3 4 1 0 2 6 9 0 2 4 9 2 3 5 7 0 3 5 1 8 5 6 9 0 Tumors 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma X 1 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hepatocellular adenoma X 1 

Mesentery + + + + + + 11 

Lipoma X 1 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Tongue + 1 

Squamous cell papilloma X 1 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Schwannoma malignant 1 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pheochromocytoma malignant X X X 4 

Pheochromocytoma benign X X X X 7 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adenoma X X X X 4 

Carcinoma X 1 

Parathyroid gland + + M + + + + + + + + + + + + + + + + + + + + + + 47 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pars distalis, adenoma X X X X X X X X X X 20 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

C-cell, adenoma X X X 5 

C-cell, adenoma, multiple 1 

Follicular cell, carcinoma X X 3 

General Body System 

Peritoneum 2 

Genital System 

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Preputial gland + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Adenoma X X X 5 

Carcinoma X X X X 5 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

62 D&C Yellow No. 11, NTP TR 463 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

Number of Days on Study 

4 

8 

1 

5 

0 

9 

5 

3 

0 

5 

9 

0 

6 

0 

0 

6 

1 

5 

6 

1 

9 

6 

2 

1 

6 

2 

2 

6 

2 

3 

6 

2 

8 

6 

3 

2 

6 

3 

2 

6 

3 

7 

6 

3 

8 

6 

5 

0 

6 

5 

6 

6 

5 

6 

6 

6 

7 

6 

7 

7 

6 

8 

8 

6 

9 

3 

6 

9 

5 

6 

9 

5 

7 

0 

1 

Carcass ID Number 

0 

5 

3 

0 

1 

1 

0 

0 

1 

0 

5 

4 

0 

4 

4 

0 

5 

8 

0 

2 

8 

0 

3 

9 

0 

4 

5 

0 

4 

0 

0 

2 

9 

0 

0 

4 

0 

3 

4 

0 

1 

5 

0 

4 

7 

0 

1 

8 

0 

3 

2 

0 

5 

0 

0 

0 

5 

0 

1 

6 

0 

3 

6 

0 

1 

7 

0 

0 

7 

0 

3 

7 

0 

0 

8 

Genital System (continued)
 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + +
 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + +
 

Testes + + + + + + M + + + + + + + + + + + + + + + + + +
 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X 

Interstitial cell, adenoma X X X X 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + 

Lymph node + + + + + + + + + + + + + 

Renal, pheochromocytoma malignant,

 metastatic, adrenal medulla X 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + 

Thymus + M + + + + + + + + + + + + + + + + + + + + + + + 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + M + + + M + + + + + 

Fibroadenoma 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 

Basal cell adenoma X 

Keratoacanthoma X X X X X 

Squamous cell papilloma 

Trichoepithelioma 

Sebaceous gland, adenoma 

Sebaceous gland, carcinoma X 

Subcutaneous tissue, fibroma X X 

Subcutaneous tissue, schwannoma malignant X 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma X 

Skeletal muscle + 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 

Hemangioma 

Peripheral nerve + 

Spinal cord + 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma X X 

Alveolar/bronchiolar carcinoma 

Osteosarcoma, metastatic, bone X 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 63 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

Number of Days on Study 

7 

0 

2 

7 

0 

6 

7 

0 

6 

7 

1 

6 

7 

1 

9 

7 

2 

2 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

4 

7 

3 

4 

Carcass ID Number 

0 

4 

3 

0 

0 

3 

0 

2 

4 

0 

3 

1 

0 

2 

0 

0 

5 

2 

0 

0 

6 

0 

0 

9 

0 

1 

0 

0 

1 

2 

0 

1 

4 

0 

1 

9 

0 

2 

2 

0 

2 

3 

0 

2 

5 

0 

2 

7 

0 

3 

0 

0 

3 

3 

0 

3 

5 

0 

4 

1 

0 

4 

8 

0 

5 

5 

0 

5 

6 

0 

5 

9 

0 

6 

0 

Total 

Tissues/ 

Tumors 

Genital System (continued) 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Testes + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X X X X X X X 30 

Interstitial cell, adenoma X X X X X 9 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node + + + + + + + 20 

Renal, pheochromocytoma malignant,

 metastatic, adrenal medulla 1 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Thymus + + + + + + + + + + + + + + + + + + M + + + + + + 48 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 48 

Fibroadenoma X X X 3 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Basal cell adenoma 1 

Keratoacanthoma X X 7 

Squamous cell papilloma X X X 3 

Trichoepithelioma X 1 

Sebaceous gland, adenoma X 1 

Sebaceous gland, carcinoma 1 

Subcutaneous tissue, fibroma X 3 

Subcutaneous tissue, schwannoma malignant 1 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Osteosarcoma 1 

Skeletal muscle 1 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hemangioma X 1 

Peripheral nerve 1 

Spinal cord 1 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Alveolar/bronchiolar adenoma X X X X X 7 

Alveolar/bronchiolar carcinoma X 1 

Osteosarcoma, metastatic, bone 1 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 50 
 

 

 

 

64 D&C Yellow No. 11, NTP TR 463 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

Number of Days on Study 

4 

8 

1 

5 

0 

9 

5 

3 

0 

5 

9 

0 

6 

0 

0 

6 

1 

5 

6 

1 

9 

6 

2 

1 

6 

2 

2 

6 

2 

3 

6 

2 

8 

6 

3 

2 

6 

3 

2 

6 

3 

7 

6 

3 

8 

6 

5 

0 

6 

5 

6 

6 

5 

6 

6 

6 

7 

6 

7 

7 

6 

8 

8 

6 

9 

3 

6 

9 

5 

6 

9 

5 

7 

0 

1 

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

Carcass ID Number 5 1 0 5 4 5 2 3 4 4 2 0 3 1 4 1 3 5 0 1 3 1 0 3 0 

3 1 1 4 4 8 8 9 5 0 9 4 4 5 7 8 2 0 5 6 6 7 7 7 8 

Special Senses System 

Ear 

Eye 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma, metastatic, bone X 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Leukemia mononuclear X X X X X X X X X X X X X X X X X X X X X 

Lymphoma malignant X 

Mesothelioma malignant X X 
 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 65 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

Number of Days on Study 

7 

0 

2 

7 

0 

6 

7 

0 

6 

7 

1 

6 

7 

1 

9 

7 

2 

2 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

4 

7 

3 

4 

Carcass ID Number 

0 

4 

3 

0 

0 

3 

0 

2 

4 

0 

3 

1 

0 

2 

0 

0 

5 

2 

0 

0 

6 

0 

0 

9 

0 

1 

0 

0 

1 

2 

0 

1 

4 

0 

1 

9 

0 

2 

2 

0 

2 

3 

0 

2 

5 

0 

2 

7 

0 

3 

0 

0 

3 

3 

0 

3 

5 

0 

4 

1 

0 

4 

8 

0 

5 

5 

0 

5 

6 

0 

5 

9 

0 

6 

0 

Total 

Tissues/ 

Tumors 

Special Senses System 

Ear 

Eye 

+ +

+

 2 

1 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Osteosarcoma, metastatic, bone 1 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Leukemia mononuclear X X X X X X X X X X X X X X X X 37 

Lymphoma malignant 1 

Mesothelioma malignant 2 
 

 

 

 

 

 

 

 

66 D&C Yellow No. 11, NTP TR 463 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm 

2 5 5 5 5 5 5 5 5 5 5 5 5 6 6 6 6 6 6 6 6 6 6 6 6 

Number of Days on Study 2 0 0 1 4 4 5 5 6 7 7 8 9 0 1 2 2 4 5 5 5 7 7 8 9 

2 9 9 0 5 6 8 8 0 0 5 6 3 2 1 8 8 2 6 6 8 2 7 8 5 

1 0 0 1 1 0 0 0 0 0 0 1 0 1 0 0 1 0 1 1 0 0 0 0 0 

Carcass ID Number 0 7 7 1 0 7 6 9 9 9 9 0 7 1 6 9 0 6 0 2 6 8 8 8 9 

1 0 5 7 4 3 4 2 7 9 1 6 4 6 2 4 8 6 3 0 1 6 8 7 0 

Alimentary System 

Esophagus + M + + + + + + + + + + + + + + + + + M + + + + + 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, cecum + + + + + + + + + + + + + M + + + + + + + + + + + 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + M + + + + + + + + + + + + + 

Intestine small, ileum + + + + + + + + + + + M + + + + + + + + + + + + + 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Hepatocellular adenoma X 

Mesentery + + + + 

Oral mucosa + 

Squamous cell carcinoma 

Squamous cell papilloma X 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Acinar cell, adenoma 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Squamous cell papilloma 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Tooth 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 

Pheochromocytoma malignant 

Pheochromocytoma benign X X X X X 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X X 

Parathyroid gland + M + + + + M M + + + + + + + + + + + + + + + + + 

Pituitary gland M + + + + + + + + + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X X X 

Pars distalis, adenoma, multiple 

Pars intermedia, adenoma 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

C-cell, adenoma X 

C-cell, adenoma, multiple 

Follicular cell, carcinoma X 

General Body System 

Peritoneum 

Genital System 

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + + 

Preputial gland + + + + + + + + + + + + + + + + + + + + + M + + + 

Adenoma 

Carcinoma X X X 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 67 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 1 1 1 1 2 2 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 

2 6 9 9 6 7 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 4 4 

1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 Total 

Carcass ID Number 1 8 8 9 9 7 6 6 6 7 7 7 8 8 8 9 9 0 0 0 0 1 1 1 1 1 Tissues/ 

0 4 0 8 6 8 5 7 8 2 7 9 1 2 9 3 5 0 5 7 9 2 3 4 5 9 Tumors 

Alimentary System 

Esophagus + + + + + M + + + + + + + + + + + + + + + + + + + + 48 

Intestine large, colon + + + + + + + + + + + I + + + + + + + + + + + + + + 50 

Intestine large, rectum + + + + + + + + + + + + + I I + + + + + + + + + + + 49 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Hepatocellular adenoma X 2 

Mesentery + + + + + + + + 12 

Oral mucosa + 2 

Squamous cell carcinoma X 1 

Squamous cell papilloma 1 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Acinar cell, adenoma X 1 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Squamous cell papilloma X X 2 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Carcinoma X 1 

Tooth + 1 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adrenal medulla + + + + + + + + + + + + + + + M + + + + + + + + + + 50 

Pheochromocytoma malignant X 1 

Pheochromocytoma benign X X X 8 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adenoma X 3 

Parathyroid gland + + + + + + + + + + + + M + + + + + + + + + + + + + 47 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pars distalis, adenoma X X X 7 

Pars distalis, adenoma, multiple X 1 

Pars intermedia, adenoma X 1 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

C-cell, adenoma X 2 

C-cell, adenoma, multiple X 1 

Follicular cell, carcinoma 1 

General Body System 

Peritoneum + 1 

Genital System 

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Preputial gland + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adenoma X X 2 

Carcinoma X 4 
 

 

 

 

 

 

 

 

 

 

 

68 D&C Yellow No. 11, NTP TR 463 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm (continued)
 

Number of Days on Study 

2 

2 

2 

5 

0 

9 

5 

0 

9 

5 

1 

0 

5 

4 

5 

5 

4 

6 

5 

5 

8 

5 

5 

8 

5 

6 

0 

5 

7 

0 

5 

7 

5 

5 

8 

6 

5 

9 

3 

6 

0 

2 

6 

1 

1 

6 

2 

8 

6 

2 

8 

6 

4 

2 

6 

5 

6 

6 

5 

6 

6 

5 

8 

6 

7 

2 

6 

7 

7 

6 

8 

8 

6 

9 

5 

Carcass ID Number 

1 

0 

1 

0 

7 

0 

0 

7 

5 

1 

1 

7 

1 

0 

4 

0 

7 

3 

0 

6 

4 

0 

9 

2 

0 

9 

7 

0 

9 

9 

0 

9 

1 

1 

0 

6 

0 

7 

4 

1 

1 

6 

0 

6 

2 

0 

9 

4 

1 

0 

8 

0 

6 

6 

1 

0 

3 

1 

2 

0 

0 

6 

1 

0 

8 

6 

0 

8 

8 

0 

8 

7 

0 

9 

0 

Genital System (continued) 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenocarcinoma X 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + + 

Testes + + + + + + + + + + + + + + + + + + + + + + + + + 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X X X 

Interstitial cell, adenoma X X X X X X 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node + + + + + + + + + + + + +
 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + M
 

Lymph node, mediastinal +
 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + +
 

Thymus + M + + + + + + + + + + + + + + + M + + + + + + +
 

Integumentary System 

Mammary gland M + + + + M + + + + + + + + + + M + + + + + + + + 

Fibroadenoma 

Skin + + + + + + + I + + + + + + + + + + + + + + + + + 

Basal cell adenoma X 

Keratoacanthoma X 

Squamous cell papilloma 

Subcutaneous tissue, fibroma X 

Subcutaneous tissue, lipoma 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Skeletal muscle + 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + +
 

Peripheral nerve +
 

Spinal cord +
 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar adenoma, multiple 

Alveolar/bronchiolar carcinoma X 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 

Special Senses System 

Ear 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Leukemia mononuclear X X X X X X X X X X X X X X X X X X X X X 

Mesothelioma malignant 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 69 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm (continued)
 

Number of Days on Study 

7 

1 

2 

7 

1 

6 

7 

1 

9 

7 

1 

9 

7 

2 

6 

7 

2 

7 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

4 

7 

3 

4 

Carcass ID Number 

1 

1 

0 

0 

8 

4 

0 

8 

0 

0 

9 

8 

0 

9 

6 

0 

7 

8 

0 

6 

5 

0 

6 

7 

0 

6 

8 

0 

7 

2 

0 

7 

7 

0 

7 

9 

0 

8 

1 

0 

8 

2 

0 

8 

9 

0 

9 

3 

0 

9 

5 

1 

0 

0 

1 

0 

5 

1 

0 

7 

1 

0 

9 

1 

1 

2 

1 

1 

3 

1 

1 

4 

1 

1 

5 

1 

1 

9 

Total 

Tissues/ 

Tumors 

Genital System (continued) 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adenocarcinoma 1 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Testes + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X X X X X X X X X X X 36 

Interstitial cell, adenoma X X X X 10 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node + + + + + + + + + + 23 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node, mediastinal 1 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Thymus + + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + + 48 

Fibroadenoma X X 2 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Basal cell adenoma 1 

Keratoacanthoma X X X 4 

Squamous cell papilloma X X X 3 

Subcutaneous tissue, fibroma X 2 

Subcutaneous tissue, lipoma X 1 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Skeletal muscle + 2 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Peripheral nerve + 2 

Spinal cord + 2 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Alveolar/bronchiolar adenoma X 1 

Alveolar/bronchiolar adenoma, multiple X 1 

Alveolar/bronchiolar carcinoma 1 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Special Senses System 

Ear + 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Leukemia mononuclear X X X X X X X X X X X X X X X 36 

Mesothelioma malignant X X 2 

1 
 

 

 

 

 

 

 

 

 

 

 

 

70 D&C Yellow No. 11, NTP TR 463 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm 

2 3 4 4 4 4 4 5 5 5 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 

Number of Days on Study 4 8 0 5 6 9 9 0 5 9 0 0 0 1 1 3 4 4 4 4 4 5 5 5 6 

6 4 6 3 8 1 9 2 1 0 0 0 7 0 5 1 6 6 9 9 9 0 0 9 3 

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

Carcass ID Number 7 7 7 3 5 2 8 7 4 7 4 6 6 3 6 6 4 6 2 3 7 6 7 3 3 

5 9 0 9 7 6 0 8 5 3 6 3 6 6 2 8 7 5 8 4 4 1 1 8 1 

Alimentary System 

Esophagus + + + + + + + + + + + + + + I + M + + + + + + + + 

Intestine large, colon + + + + + + M + + + + + + + + + + + + + + + + + + 

Polyp adenomatous 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Hepatocellular adenoma X 

Mesentery + + + + + 

Oral mucosa + 

Squamous cell carcinoma 

Squamous cell papilloma X 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Acinar cell, adenoma X 

Salivary glands + + + + + + + + + + + + + + + + M + + + + + + + + 

Carcinoma, metastatic, Zymbal’s gland X 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Tongue 

Squamous cell papilloma 

Tooth 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 

Neuroblastoma malignant X 

Pheochromocytoma malignant 

Pheochromocytoma benign X X X X X 

Bilateral, pheochromocytoma benign X 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Parathyroid gland + + + + + + + M + + + + + + + + M + + + + M + + + 

Pituitary gland + + + + + + + + M + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X X X X X X 

Pars distalis, adenoma, multiple 

Pars intermedia, adenoma 

Thyroid gland + + + + + + + + + + + + + + + + M + + + + + + + + 

C-cell, adenoma X 

C-cell, carcinoma 

Follicular cell, adenoma X 

Follicular cell, carcinoma 

General Body System 

Peritoneum + + 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 71 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

6 6 6 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 6 6 7 8 8 8 8 8 9 9 9 9 9 0 1 1 1 2 3 3 3 3 3 3 3 3 

3 7 2 0 6 6 6 6 2 5 5 5 9 6 2 4 9 7 3 3 3 3 3 3 3 3 

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Total 

Carcass ID Number 5 5 7 4 3 4 5 5 6 4 5 7 2 6 4 3 5 2 2 2 2 3 4 5 6 6 Tissues/ 

1 5 6 8 3 2 3 4 7 4 9 7 7 9 0 0 2 3 1 5 9 7 9 0 0 4 Tumors 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Polyp adenomatous X 1 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine large, cecum + + M + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, ileum + + + M + + + + + + + + + + + + + + + + + + + + + + 50 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Hepatocellular adenoma 1 

Mesentery + + + + 9 

Oral mucosa + + 3 

Squamous cell carcinoma X 1 

Squamous cell papilloma 1 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Acinar cell, adenoma X X X 4 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma, metastatic, Zymbal’s gland 1 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Tongue + 1 

Squamous cell papilloma X 1 

Tooth + + + 3 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Neuroblastoma malignant 1 

Pheochromocytoma malignant X X 2 

Pheochromocytoma benign X X X X X X 11 

Bilateral, pheochromocytoma benign 1 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Carcinoma X X 2 

Parathyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + + 48 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pars distalis, adenoma X X X X X X 13 

Pars distalis, adenoma, multiple X 1 

Pars intermedia, adenoma X 1 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

C-cell, adenoma X X 3 

C-cell, carcinoma X 1 

Follicular cell, adenoma 1 

Follicular cell, carcinoma X 1 

General Body System 

Peritoneum + 3 
 

 

 

 

 

 

 

 

 

72 D&C Yellow No. 11, NTP TR 463 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

2 3 4 4 4 4 4 5 5 5 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 

Number of Days on Study 4 8 0 5 6 9 9 0 5 9 0 0 0 1 1 3 4 4 4 4 4 5 5 5 6 

6 4 6 3 8 1 9 2 1 0 0 0 7 0 5 1 6 6 9 9 9 0 0 9 3 

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

Carcass ID Number 7 7 7 3 5 2 8 7 4 7 4 6 6 3 6 6 4 6 2 3 7 6 7 3 3 

5 9 0 9 7 6 0 8 5 3 6 3 6 6 2 8 7 5 8 4 4 1 1 8 1 

Genital System 

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + + 

Preputial gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma 

Carcinoma X 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + + 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + + 

Testes + + + + + + + + + + + + + + + + + + + + + + + + + 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X X X X X X X X 

Interstitial cell, adenoma X X X 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + 

Lymph node + + + + + + + + + + + + + + 

Lymph node, mandibular + + + + + + + + + + + + + + + + M + + + + + + + + 

Carcinoma, metastatic, Zymbal’s gland X 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + 

Hemangiosarcoma 

Thymus + + + + + + + + + + + + + + + + + + + + + + + + + 

Integumentary System 

Mammary gland + + + + + + + + + + + M + + + + + + + + + + + + + 

Fibroadenoma 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 

Basal cell adenoma X 

Basal cell carcinoma X 

Keratoacanthoma X 

Squamous cell papilloma 

Subcutaneous tissue, fibroma 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Skeletal muscle + + + + 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma X 

Carcinoma, metastatic, Zymbal’s gland X 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + M + + + + + + + + 

Special Senses System 

Ear 

Zymbal’s gland + 

Carcinoma X 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 73 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

6 6 6 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 6 6 7 8 8 8 8 8 9 9 9 9 9 0 1 1 1 2 3 3 3 3 3 3 3 3 

3 7 2 0 6 6 6 6 2 5 5 5 9 6 2 4 9 7 3 3 3 3 3 3 3 3 

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Total 

Carcass ID Number 5 5 7 4 3 4 5 5 6 4 5 7 2 6 4 3 5 2 2 2 2 3 4 5 6 6 Tissues/ 

1 5 6 8 3 2 3 4 7 4 9 7 7 9 0 0 2 3 1 5 9 7 9 0 0 4 Tumors 

Genital System 

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Preputial gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adenoma X 1 

Carcinoma 1 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Testes + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X X X X X X X X X X X X 42 

Interstitial cell, adenoma X X X 6 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node + + + + + + + + + + + + 26 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma, metastatic, Zymbal’s gland 1 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Hemangiosarcoma X 1 

Thymus + + + + M + + M + + + + + + + + + + + + + + + + + + 49 

Integumentary System 

Mammary gland + M + + + + + + + + + + + + + + + + + + + + + + + + 49 

Fibroadenoma X X X 3 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Basal cell adenoma X 2 

Basal cell carcinoma X 2 

Keratoacanthoma X 2 

Squamous cell papilloma X X 2 

Subcutaneous tissue, fibroma X X X 3 

Musculoskeletal System 

Bone 

Skeletal muscle 

+ + + + + + + + + + + + + + + + 

+

+ + + + + + + + + + 51 

5 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

+ + + + + + + + + 

+

+

+ + + + + + + + + + + + + + + + + 51 

1 

1 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Alveolar/bronchiolar adenoma X 2 

Carcinoma, metastatic, Zymbal’s gland 1 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Special Senses System 

Ear + M 1 

Zymbal’s gland 1 

Carcinoma 1 
 

 

74 D&C Yellow No. 11, NTP TR 463 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

Number of Days on Study 

2 

4 

6 

3 

8 

4 

4 

0 

6 

4 

5 

3 

4 

6 

8 

4 

9 

1 

4 

9 

9 

5 

0 

2 

5 

5 

1 

5 

9 

0 

6 

0 

0 

6 

0 

0 

6 

0 

7 

6 

1 

0 

6 

1 

5 

6 

3 

1 

6 

4 

6 

6 

4 

6 

6 

4 

9 

6 

4 

9 

6 

4 

9 

6 

5 

0 

6 

5 

0 

6 

5 

9 

6 

6 

3 

Carcass ID Number 

1 

7 

5 

1 

7 

9 

1 

7 

0 

1 

3 

9 

1 

5 

7 

1 

2 

6 

1 

8 

0 

1 

7 

8 

1 

4 

5 

1 

7 

3 

1 

4 

6 

1 

6 

3 

1 

6 

6 

1 

3 

6 

1 

6 

2 

1 

6 

8 

1 

4 

7 

1 

6 

5 

1 

2 

8 

1 

3 

4 

1 

7 

4 

1 

6 

1 

1 

7 

1 

1 

3 

8 

1 

3 

1 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Renal tubule, carcinoma 

Urethra 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Leukemia mononuclear X X X X X X X X X X X X 

Mesothelioma malignant X X 
 

 

 

 

D&C Yellow No. 11, NTP TR 463 75 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

Number of Days on Study 

6 

6 

3 

6 

6 

7 

6 

7 

2 

6 

8 

0 

6 

8 

6 

6 

8 

6 

6 

8 

6 

6 

8 

6 

6 

9 

2 

6 

9 

5 

6 

9 

5 

6 

9 

5 

6 

9 

9 

7 

0 

6 

7 

1 

2 

7 

1 

4 

7 

1 

9 

7 

2 

7 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

7 

3 

3 

Carcass ID Number 

1 

5 

1 

1 

5 

5 

1 

7 

6 

1 

4 

8 

1 

3 

3 

1 

4 

2 

1 

5 

3 

1 

5 

4 

1 

6 

7 

1 

4 

4 

1 

5 

9 

1 

7 

7 

1 

2 

7 

1 

6 

9 

1 

4 

0 

1 

3 

0 

1 

5 

2 

1 

2 

3 

1 

2 

1 

1 

2 

5 

1 

2 

9 

1 

3 

7 

1 

4 

9 

1 

5 

0 

1 

6 

0 

1 

6 

4 

Total 

Tissues/ 

Tumors 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Renal tubule, carcinoma X 1 

Urethra + 1 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Leukemia mononuclear X X X X X X X X 20 

Mesothelioma malignant X 3 
 

 

 

 

 

 

 

 

 

 

 

76 D&C Yellow No. 11, NTP TR 463 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm 

3 3 3 3 3 4 4 4 4 4 4 4 5 5 5 5 5 5 5 5 5 5 5 5 5 

Number of Days on Study 0 2 4 4 7 3 8 9 9 9 9 9 0 0 0 1 3 4 6 6 6 7 8 9 9 

2 0 5 5 0 2 1 2 6 8 8 8 2 5 9 0 4 6 3 6 8 2 9 0 6 

2 1 1 2 2 2 2 1 2 1 1 2 2 2 2 2 1 1 2 1 1 1 2 2 1 

Carcass ID Number 3 9 9 2 4 2 1 8 0 8 9 0 2 2 2 3 8 9 3 9 9 8 2 3 8 

0 7 8 7 0 4 9 7 1 6 5 3 6 1 8 8 2 2 3 3 6 4 9 4 5 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 

Hemangioma 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Hepatocellular adenoma X X 

Hepatocellular adenoma, multiple 

Mesentery + + + + + 

Oral mucosa + + 

Squamous cell carcinoma X 

Squamous cell papilloma X 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Acinar cell, adenoma 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Tongue 

Squamous cell papilloma 

Cardiovascular System 

Blood vessel + + M + + + + + + + + + + + + + + + + + + + + + + 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 

Pheochromocytoma benign X 

Bilateral, pheochromocytoma benign 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 

Parathyroid gland + + + + + + + + + + + + + + + M + M + + + + + + + 

Pituitary gland + + M M + + + + + + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X 

Pars distalis, adenoma, multiple X X 

Pars intermedia, adenoma 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

C-cell, adenoma X 

C-cell, carcinoma 

Follicular cell, carcinoma X 

General Body System 

Peritoneum + 

Genital System 

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + + 

Preputial gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 77 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

5 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 9 0 1 1 2 2 2 5 5 5 7 7 9 9 9 9 9 9 0 0 0 0 1 1 2 2 2 3 3 

6 2 5 7 4 4 9 6 6 8 8 8 0 1 3 5 5 5 0 0 4 6 6 6 2 6 7 3 3 

1 2 2 2 1 1 2 1 2 2 2 2 2 2 2 1 2 2 2 2 2 2 2 2 2 2 2 2 2 Total 

Carcass ID Number 8 1 1 0 8 9 3 9 1 1 3 3 3 1 0 9 2 3 0 0 1 2 0 0 1 1 2 0 2 Tissues/ 

8 7 0 4 9 0 7 9 6 2 2 5 9 5 8 4 0 6 7 9 4 3 5 6 1 8 2 0 5 Tumors 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Hemangioma X 1 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Carcinoma X 1 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Carcinoma X 1 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Hepatocellular adenoma X X X X 6 

Hepatocellular adenoma, multiple X 1 

Mesentery + + + + + + + + 13 

Oral mucosa + 3 

Squamous cell carcinoma 1 

Squamous cell papilloma X 2 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Acinar cell, adenoma X 1 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Tongue + + + + 4 

Squamous cell papilloma X X 2 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 53 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Pheochromocytoma benign X X X X X 6 

Bilateral, pheochromocytoma benign X X 2 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Parathyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 52 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 52 

Pars distalis, adenoma X X X X X X 8 

Pars distalis, adenoma, multiple 2 

Pars intermedia, adenoma X 1 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

C-cell, adenoma X X 3 

C-cell, carcinoma X 1 

Follicular cell, carcinoma X X 3 

General Body System 

Peritoneum + + 3 

Genital System 

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Preputial gland + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Carcinoma X X 2 
 

 

 

 

 

 

 

 

78 D&C Yellow No. 11, NTP TR 463 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

Number of Days on Study 

3 

0 

2 

3 

2 

0 

3 

4 

5 

3 

4 

5 

3 

7 

0 

4 

3 

2 

4 

8 

1 

4 

9 

2 

4 

9 

6 

4 

9 

8 

4 

9 

8 

4 

9 

8 

5 

0 

2 

5 

0 

5 

5 

0 

9 

5 

1 

0 

5 

3 

4 

5 

4 

6 

5 

6 

3 

5 

6 

6 

5 

6 

8 

5 

7 

2 

5 

8 

9 

5 

9 

0 

5 

9 

6 

Carcass ID Number 

2 

3 

0 

1 

9 

7 

1 

9 

8 

2 

2 

7 

2 

4 

0 

2 

2 

4 

2 

1 

9 

1 

8 

7 

2 

0 

1 

1 

8 

6 

1 

9 

5 

2 

0 

3 

2 

2 

6 

2 

2 

1 

2 

2 

8 

2 

3 

8 

1 

8 

2 

1 

9 

2 

2 

3 

3 

1 

9 

3 

1 

9 

6 

1 

8 

4 

2 

2 

9 

2 

3 

4 

1 

8 

5 

Genital System (continued)
 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + +
 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + +
 

Testes + + + + + + + + + + + + + + + + + + + + + + + + +
 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X X X 

Interstitial cell, adenoma X X X X 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mandibular + + + M + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mediastinal + +
 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + +
 

Thymus + + + + + + + + + + + + + + + + + + + + + + + + +
 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Fibroadenoma 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 

Keratoacanthoma X 

Squamous cell papilloma 

Sebaceous gland, carcinoma 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Skeletal muscle + + + + + + + + + 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 

Special Senses System 

Ear 

Eye + 

Zymbal’s gland 

Carcinoma 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Renal tubule, adenoma 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Leukemia mononuclear X X X X X X X 

Mesothelioma malignant X X 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 79 

TABLE A2
 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

Number of Days on Study 

5 

9 

6 

6 

0 

2 

6 

1 

5 

6 

1 

7 

6 

2 

4 

6 

2 

4 

6 

2 

9 

6 

5 

6 

6 

5 

6 

6 

5 

8 

6 

7 

8 

6 

7 

8 

6 

9 

0 

6 

9 

1 

6 

9 

3 

6 

9 

5 

6 

9 

5 

6 

9 

5 

7 

0 

0 

7 

0 

0 

7 

0 

4 

7 

0 

6 

7 

1 

6 

7 

1 

6 

7 

2 

2 

7 

2 

6 

7 

2 

7 

7 

3 

3 

7 

3 

3 

Carcass ID Number 

1 

8 

8 

2 

1 

7 

2 

1 

0 

2 

0 

4 

1 

8 

9 

1 

9 

0 

2 

3 

7 

1 

9 

9 

2 

1 

6 

2 

1 

2 

2 

3 

2 

2 

3 

5 

2 

3 

9 

2 

1 

5 

2 

0 

8 

1 

9 

4 

2 

2 

0 

2 

3 

6 

2 

0 

7 

2 

0 

9 

2 

1 

4 

2 

2 

3 

2 

0 

5 

2 

0 

6 

2 

1 

1 

2 

1 

8 

2 

2 

2 

2 

0 

0 

2 

2 

5 

Total 

Tissues/ 

Tumors 

Genital System (continued) 

Prostate + + + + + + + + + + + + + + + + + + + + + + + + + + + + 53 

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Testes + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Bilateral, interstitial cell, adenoma X X X X X X X X X X X X X X X X X X X X X X X X X 39 

Interstitial cell, adenoma X X X 7 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Lymph node + + + + + + + + + + + + + + + + + + + + + + 45 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 53 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Lymph node, mediastinal 2 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Thymus M + + + + + + + + + + + + + + + + + + + + + + + + + + + + 53 

Integumentary System 

Mammary gland + + + + M + + + + + + + + + + + + + + + + + + + + + + + + 53 

Fibroadenoma X 1 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Keratoacanthoma 1 

Squamous cell papilloma X X X 3 

Sebaceous gland, carcinoma X 1 

Musculoskeletal System 

Bone 

Skeletal muscle 

+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

9 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Alveolar/bronchiolar adenoma X 1 

Alveolar/bronchiolar carcinoma X 1 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Special Senses System 

Ear + 1 

Eye 1 

Zymbal’s gland + 1 

Carcinoma X 1 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Renal tubule, adenoma X X 2 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + + + + + 54 

Leukemia mononuclear X X X X X X X X X X X X X X X 22 

Mesothelioma malignant X X 4 
80 D&C Yellow No. 11, NTP TR 463 

TABLE A3 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Adrenal Medulla: Benign Pheochromocytoma a Overall rate 7/50 (14%) 8/50 (16%) 12/51 (24%) 8/54 (15%) b Adjusted rate 25.9% 25.8% 49.3% 55.7% c Terminal rate 3/19 (16%) 1/19 (5%) 1/8 (13%) 0/2 (0%) 

First incidence (days) 615 560 499 498 

d Life table test P=0.025 P=0.483 P=0.028 P=0.032 

d Logistic regression test P=0.377 P=0.469 P=0.143 P=0.258 

d Cochran-Armitage test P=0.533N 

d Fisher exact test P=0.500 P=0.166 P=0.565 

Adrenal Medulla: Malignant Pheochromocytoma 

Overall rate 4/50 (8%) 1/50 (2%) 2/51 (4%) 0/54 (0%) 

Adjusted rate 18.5% 5.3% 19.8% 0.0% 

Terminal rate 3/19 (16%) 1/19 (5%) 1/8 (13%) 0/2 (0%) e First incidence (days) 677 733 (T) 712 —

Life table test P=0.454N P=0.182N P=0.663 P=0.404N 

Logistic regression test P=0.255N P=0.186N P=0.574N P=0.211N 

Cochran-Armitage test P=0.072N 

Fisher exact test P=0.181N P=0.329N P=0.050N 

Adrenal Medulla: Benign or Malignant Pheochromocytoma 

Overall rate 10/50 (20%) 9/50 (18%) 14/51 (27%) 8/54 (15%) 

Adjusted rate 39.8% 29.9% 60.2% 55.7% 

Terminal rate 6/19 (32%) 2/19 (11%) 2/8 (25%) 0/2 (0%) 

First incidence (days) 615 560 499 498 

Life table test P=0.033 P=0.516N P=0.024 P=0.051 

Logistic regression test P=0.521 P=0.558N P=0.167 P=0.456 

Cochran-Armitage test P=0.286N 

Fisher exact test P=0.500N P=0.260 P=0.330N 

Kidney (Renal Tubule): Adenoma (Step Sections) 

Overall rate 0/50 (0%) 2/51 (4%) 4/51 (8%) 2/54 (4%) 

Adjusted rate 0.0% 7.1% 22.5% 18.8% 

Terminal rate 0/19 (0%) 1/20 (5%) 1/8 (13%) 0/2 (0%) 

First incidence (days) — 558 649 678 

Life table test P=0.080 P=0.241 P=0.027 P=0.090 

Logistic regression test P=0.259 P=0.255 P=0.046 P=0.120 

Cochran-Armitage test P=0.406 

Fisher exact test P=0.252 P=0.061 P=0.267 

Kidney (Renal Tubule): Adenoma (Single and Step Sections) 

Overall rate 0/50 (0%) 2/51 (4%) 4/51 (8%) 4/54 (7%) 

Adjusted rate 0.0% 7.1% 22.5% 38.3% 

Terminal rate 0/19 (0%) 1/20 (5%) 1/8 (13%) 0/2 (0%) 

First incidence (days) — 558 649 658 

Life table test P=0.003 P=0.241 P=0.027 P=0.006 

Logistic regression test P=0.032 P=0.255 P=0.046 P=0.014 

Cochran-Armitage test P=0.106 

Fisher exact test P=0.252 P=0.061 P=0.069 
D&C Yellow No. 11, NTP TR 463 81 

TABLE A3
 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Kidney (Renal Tubule): Adenoma or Carcinoma (Single and Step Sections) 

Overall rate 0/50 (0%) 2/51 (4%) 5/51 (10%) 4/54 (7%) 

Adjusted rate 0.0% 7.1% 26.1% 38.3% 

Terminal rate 0/19 (0%) 1/20 (5%) 1/8 (13%) 0/2 (0%) 

First incidence (days) — 558 649 658 

Life table test P=0.004 P=0.241 P=0.012 P=0.006 

Logistic regression test P=0.036 P=0.255 P=0.022 P=0.014 

Cochran-Armitage test P=0.121 

Fisher exact test P=0.252 P=0.030 P=0.069 

Liver: Hepatocellular Adenoma 

Overall rate 1/50 (2%) 2/51 (4%) 1/51 (2%) 7/54 (13%) 

Adjusted rate 5.3% 7.9% 2.6% 75.0% 

Terminal rate 1/19 (5%) 1/20 (5%) 0/8 (0%) 1/2 (50%) 

First incidence (days) 733 (T) 656 607 498 

Life table test P<0.001 P=0.508 P=0.650 P<0.001 

Logistic regression test P=0.001 P=0.487 P=0.757 P=0.008 

Cochran-Armitage test P=0.007 

Fisher exact test P=0.508 P=0.748N P=0.038 

Lung: Alveolar/bronchiolar Adenoma 

Overall rate 7/50 (14%) 2/51 (4%) 2/51 (4%) 1/54 (2%) 

Adjusted rate 28.9% 10.0% 9.6% 7.1% 

Terminal rate 4/19 (21%) 2/20 (10%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 650 733 (T) 453 695 

Life table test P=0.405N P=0.073N P=0.294N P=0.414N 

Logistic regression test P=0.112N P=0.084N P=0.103N P=0.148N 

Cochran-Armitage test P=0.045N 

Fisher exact test P=0.075N P=0.075N P=0.023N 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 8/50 (16%) 3/51 (6%) 2/51 (4%) 2/54 (4%) 

Adjusted rate 33.7% 12.0% 9.6% 20.4% 

Terminal rate 5/19 (26%) 2/20 (10%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 650 558 453 695 

Life table test P=0.574N P=0.092N P=0.233N P=0.642N 

Logistic regression test P=0.155N P=0.111N P=0.069N P=0.260N 

Cochran-Armitage test P=0.063N 

Fisher exact test P=0.094N P=0.043N P=0.035N 

Mammary Gland: Fibroadenoma 

Overall rate 3/50 (6%) 2/51 (4%) 3/51 (6%) 1/54 (2%) 

Adjusted rate 14.6% 10.0% 29.2% 20.0% 

Terminal rate 2/19 (11%) 2/20 (10%) 2/8 (25%) 0/2 (0%) 

First incidence (days) 716 733 (T) 692 722 

Life table test P=0.285 P=0.469N P=0.285 P=0.573 

Logistic regression test P=0.532 P=0.470N P=0.394 P=0.729 

Cochran-Armitage test P=0.247N 

Fisher exact test P=0.491N P=0.652N P=0.280N 
82 D&C Yellow No. 11, NTP TR 463 

TABLE A3
 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Oral Cavity (Oral Mucosa or Tongue): Squamous Cell Papilloma 

Overall rate 1/50 (2%) 1/51 (2%) 2/51 (4%) 4/54 (7%) 

Adjusted rate 5.3% 3.2% 6.5% 28.1% 

Terminal rate 1/19 (5%) 0/20 (0%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 733 (T) 658 406 481 

Life table test P=0.008 P=0.758N P=0.367 P=0.020 

Logistic regression test P=0.087 P=0.755 P=0.606 P=0.110 

Cochran-Armitage test P=0.088 

Fisher exact test P=0.748N P=0.508 P=0.206 

Oral Cavity (Oral Mucosa or Tongue): Squamous Cell Papilloma or Squamous Cell Carcinoma 

Overall rate 1/50 (2%) 2/51 (4%) 3/51 (6%) 5/54 (9%) 

Adjusted rate 5.3% 6.9% 10.6% 30.4% 

Terminal rate 1/19 (5%) 0/20 (0%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 733 (T) 658 406 481 

Life table test P=0.004 P=0.521 P=0.191 P=0.008 

Logistic regression test P=0.066 P=0.487 P=0.369 P=0.069 

Cochran-Armitage test P=0.078 

Fisher exact test P=0.508 P=0.316 P=0.121 

Pancreas: Adenoma 

Overall rate 0/50 (0%) 1/51 (2%) 4/51 (8%) 1/54 (2%) 

Adjusted rate 0.0% 5.0% 18.0% 5.3% 

Terminal rate 0/19 (0%) 1/20 (5%) 0/8 (0%) 0/2 (0%) 

First incidence (days) — 733 (T) 649 678 

Life table test P=0.236 P=0.510 P=0.031 P=0.409 

Logistic regression test P=0.368 P=0.510 P=0.048 P=0.432 

Cochran-Armitage test P=0.540 

Fisher exact test P=0.505 P=0.061 P=0.519 

Pancreatic Islets: Adenoma 

Overall rate 4/50 (8%) 3/51 (6%) 0/51 (0%) 0/54 (0%)
 

Adjusted rate 19.6% 8.9% 0.0% 0.0%
 

Terminal rate 3/19 (16%) 0/20 (0%) 0/8 (0%) 0/2 (0%)
 

First incidence (days) 716 586 — —
 

Life table test P=0.146N P=0.494N P=0.215N P=0.483N
 

Logistic regression test P=0.050N P=0.514N P=0.167N P=0.320N
 

Cochran-Armitage test P=0.028N
 

Fisher exact test P=0.489N P=0.056N P=0.050N
 

Pancreatic Islets: Adenoma or Carcinoma 

Overall rate 5/50 (10%) 3/51 (6%) 2/51 (4%) 0/54 (0%)
 

Adjusted rate 24.6% 8.9% 16.5% 0.0%
 

Terminal rate 4/19 (21%) 0/20 (0%) 1/8 (13%) 0/2 (0%)
 

First incidence (days) 716 586 686 —
 

Life table test P=0.222N P=0.350N P=0.597N P=0.430N
 

Logistic regression test P=0.066N P=0.374N P=0.479N P=0.265N
 

Cochran-Armitage test P=0.024N
 

Fisher exact test P=0.346N P=0.219N P=0.023N
 
D&C Yellow No. 11, NTP TR 463 83 

TABLE A3
 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 20/50 (40%) 8/50 (16%) 14/50 (28%) 10/52 (19%) 

Adjusted rate 63.9% 27.7% 58.0% 36.1% 

Terminal rate 10/19 (53%) 3/20 (15%) 3/8 (38%) 0/2 (0%) 

First incidence (days) 530 586 502 509 

Life table test P=0.129 P=0.014N P=0.492 P=0.342 

Logistic regression test P=0.160N P=0.008N P=0.214N P=0.046N 

Cochran-Armitage test P=0.098N 

Fisher exact test P=0.007N P=0.146N P=0.018N 

Preputial Gland: Adenoma 

Overall rate 5/49 (10%) 2/50 (4%) 1/51 (2%) 0/54 (0%) 

Adjusted rate 21.3% 9.3% 4.5% 0.0% 

Terminal rate 3/19 (16%) 1/20 (5%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 638 726 686 — 

Life table test P=0.200N P=0.203N P=0.287N P=0.302N 

Logistic regression test P=0.083N P=0.229N P=0.167N P=0.107N 

Cochran-Armitage test P=0.026N 

Fisher exact test P=0.210N P=0.093N P=0.022N 

Preputial Gland: Carcinoma 

Overall rate 5/49 (10%) 4/50 (8%) 1/51 (2%) 2/54 (4%) 

Adjusted rate 23.7% 13.4% 3.3% 35.7% 

Terminal rate 4/19 (21%) 1/20 (5%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 688 510 650 716 

Life table test P=0.548 P=0.496N P=0.318N P=0.355 

Logistic regression test P=0.327N P=0.522N P=0.192N P=0.645 

Cochran-Armitage test P=0.142N 

Fisher exact test P=0.487N P=0.093N P=0.180N 

Preputial Gland: Adenoma or Carcinoma 

Overall rate 10/49 (20%) 6/50 (12%) 2/51 (4%) 2/54 (4%) 

Adjusted rate 42.9% 21.7% 7.7% 35.7% 

Terminal rate 7/19 (37%) 2/20 (10%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 638 510 650 716 

Life table test P=0.365N P=0.199N P=0.138N P=0.638N 

Logistic regression test P=0.068N P=0.233N P=0.044N P=0.184N 

Cochran-Armitage test P=0.010N 

Fisher exact test P=0.194N P=0.011N P=0.009N 

Skin: Squamous Cell Papilloma 

Overall rate 3/50 (6%) 3/51 (6%) 2/51 (4%) 3/54 (6%) 

Adjusted rate 13.7% 15.0% 14.1% 58.0% 

Terminal rate 1/19 (5%) 3/20 (15%) 0/8 (0%) 1/2 (50%) 

First incidence (days) 706 733 (T) 686 624 

Life table test P=0.036 P=0.633N P=0.577 P=0.101 

Logistic regression test P=0.236 P=0.652N P=0.674N P=0.359 

Cochran-Armitage test P=0.571N 

Fisher exact test P=0.652N P=0.491N P=0.623N 
84 D&C Yellow No. 11, NTP TR 463 

TABLE A3
 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Skin: Keratoacanthoma 

Overall rate 7/50 (14%) 4/51 (8%) 2/51 (4%) 1/54 (2%) 

Adjusted rate 23.1% 17.7% 11.4% 3.3% 

Terminal rate 2/19 (11%) 3/20 (15%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 628 658 650 596 

Life table test P=0.244N P=0.276N P=0.232N P=0.236N 

Logistic regression test P=0.064N P=0.279N P=0.105N P=0.054N 

Cochran-Armitage test P=0.025N 

Fisher exact test P=0.251N P=0.075N P=0.023N 

Skin: Squamous Cell Papilloma or Keratoacanthoma 

Overall rate 10/50 (20%) 7/51 (14%) 4/51 (8%) 4/54 (7%) 

Adjusted rate 34.1% 32.3% 23.9% 59.4% 

Terminal rate 3/19 (16%) 6/20 (30%) 0/8 (0%) 1/2 (50%) 

First incidence (days) 628 658 650 596 

Life table test P=0.369 P=0.293N P=0.335N P=0.544 

Logistic regression test P=0.282N P=0.327N P=0.134N P=0.230N 

Cochran-Armitage test P=0.059N 

Fisher exact test P=0.282N P=0.069N P=0.055N 

Skin: Trichoepithelioma, Basal Cell Adenoma, or Basal Cell Carcinoma 

Overall rate 2/50 (4%) 1/51 (2%) 4/51 (8%) 0/54 (0%)
 

Adjusted rate 8.6% 3.3% 21.9% 0.0%
 

Terminal rate 1/19 (5%) 0/20 (0%) 1/8 (13%) 0/2 (0%)
 

First incidence (days) 695 672 590 —
 

Life table test P=0.545N P=0.502N P=0.156 P=0.551N
 

Logistic regression test P=0.322N P=0.513N P=0.280 P=0.416N
 

Cochran-Armitage test P=0.216N
 

Fisher exact test P=0.492N P=0.348 P=0.229N
 

Skin: Squamous Cell Papilloma, Keratoacanthoma, Trichoepithelioma, Basal Cell Adenoma, or Basal Cell Carcinoma 

Overall rate 12/50 (24%) 8/51 (16%) 8/51 (16%) 4/54 (7%) 

Adjusted rate 40.5% 34.5% 40.6% 59.4% 

Terminal rate 4/19 (21%) 6/20 (30%) 1/8 (13%) 1/2 (50%) 

First incidence (days) 628 658 590 596 

Life table test P=0.441 P=0.229N P=0.516 P=0.595N 

Logistic regression test P=0.171N P=0.255N P=0.359N P=0.135N 

Cochran-Armitage test P=0.025N 

Fisher exact test P=0.213N P=0.213N P=0.018N 

Skin (Subcutaneous Tissue): Fibroma 

Overall rate 3/50 (6%) 2/51 (4%) 3/51 (6%) 0/54 (0%)
 

Adjusted rate 10.1% 6.0% 23.4% 0.0%
 

Terminal rate 1/19 (5%) 0/20 (0%) 1/8 (13%) 0/2 (0%)
 

First incidence (days) 632 558 663 —
 

Life table test P=0.359N P=0.509N P=0.442 P=0.333N
 

Logistic regression test P=0.140N P=0.468N P=0.602 P=0.141N
 

Cochran-Armitage test P=0.097N
 

Fisher exact test P=0.491N P=0.652N P=0.108N
 
D&C Yellow No. 11, NTP TR 463 85 

TABLE A3
 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Testes: Adenoma 

Overall rate 39/49 (80%) 46/51 (90%) 48/51 (94%) 46/54 (85%) 

Adjusted rate 95.0% 100.0% 100.0% 100.0% 

Terminal rate 17/19 (89%) 20/20 (100%) 8/8 (100%) 2/2 (100%) 

First incidence (days) 481 509 453 370 

Life table test P<0.001 P=0.212 P<0.001 P<0.001 

Logistic regression test P=0.018 P=0.033 P<0.001 P=0.005 

Cochran-Armitage test P=0.533 

Fisher exact test P=0.114 P=0.030 P=0.313 

Thyroid Gland (C-cell): Adenoma 

Overall rate 6/50 (12%) 3/51 (6%) 3/50 (6%) 3/54 (6%) 

Adjusted rate 20.7% 12.2% 11.5% 23.8% 

Terminal rate 2/19 (11%) 2/20 (10%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 590 575 610 498 

Life table test P=0.373 P=0.265N P=0.484N P=0.525 

Logistic regression test P=0.379N P=0.242N P=0.274N P=0.325N 

Cochran-Armitage test P=0.267N 

Fisher exact test P=0.234N P=0.243N P=0.207N 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate 6/50 (12%) 3/51 (6%) 4/50 (8%) 4/54 (7%) 

Adjusted rate 20.7% 12.2% 22.5% 26.5% 

Terminal rate 2/19 (11%) 2/20 (10%) 1/8 (13%) 0/2 (0%) 

First incidence (days) 590 575 610 498 

Life table test P=0.157 P=0.265N P=0.594 P=0.350 

Logistic regression test P=0.568N P=0.242N P=0.436N P=0.445N 

Cochran-Armitage test P=0.419N 

Fisher exact test P=0.234N P=0.370N P=0.322N 

Thyroid Gland (Follicular Cell): Carcinoma 

Overall rate 3/50 (6%) 1/51 (2%) 1/50 (2%) 3/54 (6%) 

Adjusted rate 13.7% 2.2% 12.5% 37.8% 

Terminal rate 2/19 (11%) 0/20 (0%) 1/8 (13%) 0/2 (0%) 

First incidence (days) 695 558 733 (T) 596 

Life table test P=0.055 P=0.305N P=0.602N P=0.144 

Logistic regression test P=0.321 P=0.307N P=0.514N P=0.499 

Cochran-Armitage test P=0.449 

Fisher exact test P=0.301N P=0.309N P=0.623N 

Thyroid Gland (Follicular Cell): Adenoma or Carcinoma 

Overall rate 3/50 (6%) 1/51 (2%) 2/50 (4%) 3/54 (6%) 

Adjusted rate 13.7% 2.2% 15.6% 37.8% 

Terminal rate 2/19 (11%) 0/20 (0%) 1/8 (13%) 0/2 (0%) 

First incidence (days) 695 558 659 596 

Life table test P=0.063 P=0.305N P=0.590 P=0.144 

Logistic regression test P=0.333 P=0.307N P=0.662N P=0.499 

Cochran-Armitage test P=0.463 

Fisher exact test P=0.301N P=0.500N P=0.623N 
86 D&C Yellow No. 11, NTP TR 463 

TABLE A3
 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

All Organs: Mononuclear Cell Leukemia 

Overall rate 37/50 (74%) 36/51 (71%) 20/51 (39%) 22/54 (41%) 

Adjusted rate 80.0% 77.2% 61.7% 81.6% 

Terminal rate 10/19 (53%) 10/20 (50%) 2/8 (25%) 0/2 (0%) 

First incidence (days) 481 222 453 481 

Life table test P=0.191 P=0.524N P=0.241N P=0.151 

Logistic regression test P<0.001N P=0.358N P<0.001N P=0.011N 

Cochran-Armitage test P<0.001N 

Fisher exact test P=0.436N P<0.001N P<0.001N 

All Organs: Malignant Mesothelioma 

Overall rate 2/50 (4%) 2/51 (4%) 3/51 (6%) 4/54 (7%) 

Adjusted rate 6.7% 9.5% 8.7% 22.7% 

Terminal rate 0/19 (0%) 1/20 (5%) 0/8 (0%) 0/2 (0%) 

First incidence (days) 656 727 491 496 

Life table test P=0.037 P=0.681N P=0.384 P=0.127 

Logistic regression test P=0.258 P=0.682 P=0.592 P=0.363 

Cochran-Armitage test P=0.263 

Fisher exact test P=0.684N P=0.509 P=0.377 

All Organs: Benign Neoplasms 

Overall rate 49/50 (98%) 50/51 (98%) 49/51 (96%) 49/54 (91%) 

Adjusted rate 100.0% 100.0% 100.0% 100.0% 

Terminal rate 19/19 (100%) 20/20 (100%) 8/8 (100%) 2/2 (100%) 

First incidence (days) 481 509 406 370 

Life table test P<0.001 P=0.492 P=0.008 P<0.001 

Logistic regression test P=0.575 P=0.488 P=0.482 P=0.427 

Cochran-Armitage test P=0.034N 

Fisher exact test P=0.748 P=0.508N P=0.121N 

All Organs: Malignant Neoplasms 

Overall rate 42/50 (84%) 39/51 (76%) 30/51 (59%) 26/54 (48%) 

Adjusted rate 87.3% 82.1% 89.6% 88.8% 

Terminal rate 13/19 (68%) 12/20 (60%) 6/8 (75%) 0/2 (0%) 

First incidence (days) 481 222 384 481 

Life table test P=0.065 P=0.424N P=0.417 P=0.080 

Logistic regression test P<0.001N P=0.189N P=0.005N P=0.005N 

Cochran-Armitage test P<0.001N 

Fisher exact test P=0.243N P=0.005N P<0.001N 
87 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE A3
 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

All Organs: Benign or Malignant Neoplasms 

Overall rate 50/50 (100%) 51/51 (100%) 50/51 (98%) 49/54 (91%) 

Adjusted rate 100.0% 100.0% 100.0% 100.0% 

Terminal rate 19/19 (100%) 20/20 (100%) 8/8 (100%) 2/2 (100%) 

First incidence (days) 481 222 384 370 

Life table test 

Logistic regression test 

P<0.001 

P=0.018N 

P=0.487 

—f 

P=0.009 

— 

P<0.001 

P=0.973N 

Cochran-Armitage test P=0.002N 

Fisher exact test P=1.000N P=0.505N P=0.034N 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, kidney, liver, 

lung, pancreas, pancreatic islets, pituitary gland, preputial gland, testes, and thyroid gland; for other tissues, denominator is number of animals necropsied. b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality
 

Observed incidence at terminal kill
 d Beneath the control incidence are the P values associated with the trend test. Beneath the exposed group incidence are the P values corresponding to pairwise 

comparisons between the controls and that exposed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) 

the cause of death. The logistic regression test regards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence 

rates. For all tests, a negative trend or a lower incidence in an exposure group is indicated byN. e Not applicable; no neoplasms in animal group f Value of statistic cannot be computed. 
88 D&C Yellow No. 11, NTP TR 463 

TABLE A4a 

Historical Incidence of Hepatocellular Neoplasms in Untreated Male F344/N Rats a 

Incidence in Controls 

Study Adenoma Carcinoma Adenoma or Carcinoma 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 0/51 0/51 0/51 

Benzyl Acetate 5/50 1/50 5/50 

Butyl Benzyl Phthalate 2/50 0/50 2/50 

C.I. Pigment Red 23 2/50 1/50 3/50 

C.I. Pigment Red 3 0/50 0/50 0/50 

o-Nitroanisole 0/50 0/50 0/50 

p-Nitrobenzoic Acid 2/49 3/49 4/49 

Polysorbate 80 2/50 0/50 2/50 

Overall Historical Incidence 

Total 30/1,301 (2.3%) 9/1,301 (0.7%) 37/1,301 (2.8%) 

Standard deviation 2.9% 1.4% 3.3% 

Range 0%-10% 0%-6% 0%-10% 

a Data as of 12 May 1995 

TABLE A4b 

Historical Incidence of Renal Tubule Neoplasms in Untreated Male F344/N Rats a 

Incidence in Controls 

Study Adenoma Carcinoma Adenoma or Carcinoma 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 0/51 0/51 0/51 

Benzyl Acetate 0/50 0/50 0/50 

Butyl Benzyl Phthalate 1/50 0/50 1/50 

C.I. Pigment Red 23 0/50 0/50 0/50 

C.I. Pigment Red 3 0/50 1/50 1/50 

o-Nitroanisole 0/49 0/49 0/49 

p-Nitrobenzoic Acid 0/50 0/50 0/50 

Polysorbate 80 0/50 1/50 1/50 

Overall Historical Incidence 

Total 9/1,301 (0.7%) 3/1,301 (0.2%) 12/1,301 (0.9%) 

Standard deviation 1.5% 0.7% 1.5% 

Range 0%-6% 0%-2% 0%-6% 

a Data as of 12 May 1995 
D&C Yellow No. 11, NTP TR 463 89 

TABLE A4c 

Historical Incidence of Oral Cavity Neoplasms in Untreated Male F344/N Rats a 

Incidence in Controls 

Study Squamous Cell 

Papillomab 

Squamous Cell 

Carcinoma 

Squamous Cell 

Papilloma or Carcinomab 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 0/51 0/51 0/51 

Benzyl Acetate 0/50 0/50 0/50 

Butyl Benzyl Phthalate 0/50 0/50 0/50 

C.I. Pigment Red 23 0/50 0/50 0/50 

C.I. Pigment Red 3 1/50 0/50 1/50 

o-Nitroanisole 0/50 0/50 0/50 

p-Nitrobenzoic Acid 1/50 0/50 1/50 

Polysorbate 80 1/50 0/50 1/50 

Overall Historical Incidence 

Total 10/1,304 (0.8%) 0/1,304 (0%) 10/1,304 (0.8%) 

Standard deviation 1.3% 1.3% 

Range 0%-4% 0%-4% 

a Data as of 12 May 1995. Includes data for oral mucosa, tongue, pharynx, and tooth. b Includes data for papilloma. 

TABLE A4d 

Historical Incidence of Testicular Adenoma in Untreated Male F344/N Rats a 

Study Incidence in Controls 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 49/51 

Benzyl Acetate 47/50 

Butyl Benzyl Phthalate 44/50 

C.I. Pigment Red 23 48/50 

C.I. Pigment Red 3 47/50 

o-Nitroanisole 48/50 

p-Nitrobenzoic Acid 44/50 

Polysorbate 80 39/49 

Overall Historical Incidence 

Total 1,169/1,302 (89.8%) 

Standard deviation 5.9% 

Range 74%-98% 

a Data as of 12 May 1995 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

90 D&C Yellow No. 11, NTP TR 463 

TABLE A5 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 a 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Disposition Summary 

Animals initially in study 60 60 60 60 

12-Month interim evaluation 10 9 9 6 

Early deaths 

Moribund 29 29 41 49 

Natural deaths 2 2 1 3 

Other 1 

Survivors 

Terminal sacrifice 19 20 8 2 

Animals examined microscopically 60 60 60 60 

12-Month Interim Evaluation 

Alimentary System 

Intestine large, colon (9) (9) (8) (5) 

Parasite metazoan 1 (11%) 1 (13%) 

Intestine large, rectum (10) (9) (9) (6) 

Parasite metazoan 1 (10%) 2 (22%) 

Intestine large, cecum (10) (9) (9) (6) 

Parasite metazoan 1 (11%) 1 (11%) 

Liver (10) (9) (9) (6) 

Basophilic focus 1 (10%) 1 (11%) 1 (17%) 

Clear cell focus 1 (11%) 1 (17%) 

Eosinophilic focus 2 (22%) 

Granuloma 1 (10%) 

Hepatodiaphragmatic nodule 1 (10%) 1 (11%) 

Inflammation, subacute 1 (10%) 2 (22%) 2 (22%) 2 (33%) 

Mixed cell focus 1 (17%) 

Bile duct, hyperplasia 4 (40%) 1 (17%) 

Bile duct, pigmentation 9 (100%) 9 (100%) 6 (100%) 

Hepatocyte, cytologic alterations 8 (89%) 9 (100%) 6 (100%) 

Hepatocyte, pigmentation 1 (11%) 8 (89%) 6 (100%) 

Kupffer cell, pigmentation 5 (83%) 

Mesentery (1) (1) 

Fat, necrosis 1 (100%) 1 (100%) 

Oral mucosa (1) 

Hyperplasia 1 (100%) 

Pancreas (10) (9) (9) (6) 

Atrophy 2 (22%) 4 (44%) 2 (33%) 

Inflammation, chronic 1 (11%) 

Cardiovascular System 

Heart (10) (9) (9) (6) 

Cardiomyopathy 5 (50%) 3 (33%) 2 (22%) 3 (50%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 91 

TABLE A5
 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

12-Month Interim Evaluation (continued) 

Endocrine System 

Adrenal cortex (10) (9) (9) (6) 

Accessory adrenal cortical nodule 1 (11%) 1 (11%) 1 (17%) 

Hypertrophy, focal 1 (11%) 

Pituitary gland (10) (9) (8) (6) 

Pars distalis, angiectasis 1 (10%) 

Pars distalis, cyst 1 (10%) 1 (11%) 

Pars distalis, hyperplasia, focal 1 (10%) 1 (17%) 

Pars intermedia, angiectasis 1 (10%) 

Thyroid gland (10) (9) (9) (6) 

Ultimobranchial cyst 1 (10%) 2 (22%) 

Genital System 

Epididymis (10) (9) (9) (6) 

Atypia cellular 1 (11%) 

Hypospermia 1 (10%) 

Preputial gland (10) (9) (9) (6) 

Inflammation, chronic 5 (50%) 3 (33%) 2 (22%) 2 (33%) 

Prostate (10) (9) (9) (6) 

Corpora amylacea 1 (11%) 1 (11%) 1 (17%) 

Inflammation, suppurative 3 (30%) 7 (78%) 5 (56%) 4 (67%) 

Testes (10) (9) (9) (6) 

Interstitial cell, hyperplasia 7 (70%) 4 (44%) 3 (33%) 3 (50%) 

Seminiferous tubule, atrophy 2 (20%) 1 (17%) 

Hematopoietic System 

Lymph node (2) (2) (4) (2) 

Mediastinal, hemorrhage 2 (100%) 2 (100%) 2 (50%) 1 (50%) 

Mediastinal, hyperplasia, lymphoid 2 (50%) 

Mediastinal, pigmentation 2 (100%) 1 (50%) 1 (25%) 2 (100%) 

Lymph node, mandibular (10) (8) (9) (6) 

Hemorrhage 5 (50%) 2 (22%) 1 (17%) 

Hyperplasia, lymphoid 1 (13%) 

Pigmentation 1 (10%) 1 (13%) 3 (33%) 1 (17%) 

Lymph node, mesenteric (10) (9) (9) (6) 

Ectasia 2 (20%) 1 (17%) 

Hemorrhage 1 (11%) 

Respiratory System 

Lung (10) (9) (9) (6) 

Hemorrhage 1 (17%) 

Infiltration cellular, histiocyte 2 (20%) 2 (22%) 2 (22%) 1 (17%) 

Inflammation, subacute 3 (33%) 3 (50%) 

Alveolar epithelium, hyperplasia 1 (17%) 

Nose (10) (9) (9) (6) 

Mucosa, metaplasia, squamous 1 (11%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

92 D&C Yellow No. 11, NTP TR 463 

TABLE A5
 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

12-Month Interim Evaluation (continued) 

Special Senses System 

Eye

Cataract

Hemorrhage

Retina, degeneration

 (3) 

 1 (33%) 

 2 (67%) 

 1 (33%) 

Urinary System 

Kidney (10) (9) (9) (6) 

Infarct 1 (11%) 

Mineralization 2 (20%) 

Nephropathy 10 (100%) 9 (100%) 9 (100%) 6 (100%) 

Renal tubule, pigmentation 7 (78%) 9 (100%) 6 (100%) 

Systems Examined With No Lesions Observed 

General Body System 

Integumentary System 

Musculoskeletal System 

Nervous System 

2-Year Study 

Alimentary System 

Intestine large, colon (50) (50) (50) (54) 

Edema 1 (2%) 1 (2%) 

Parasite metazoan 4 (8%) 4 (8%) 8 (16%) 3 (6%) 

Intestine large, rectum (50) (49) (51) (54) 

Parasite metazoan 9 (18%) 1 (2%) 3 (6%) 8 (15%) 

Intestine large, cecum (50) (50) (50) (54) 

Edema 1 (2%) 4 (8%) 8 (16%) 12 (22%) 

Parasite metazoan 2 (4%) 1 (2%) 1 (2%) 3 (6%) 

Intestine small, duodenum (50) (51) (51) (54) 

Erosion 1 (2%) 

Epithelium, hyperplasia 1 (2%) 4 (8%) 22 (43%) 21 (39%) 

Intestine small, jejunum (50) (50) (51) (54) 

Cyst 1 (2%) 

Inflammation, chronic 2 (4%) 

Ulcer 1 (2%) 

Epithelium, hyperplasia 3 (6%) 10 (20%) 12 (22%) 

Intestine small, ileum (49) (50) (50) (54) 

Epithelium, hyperplasia 3 (6%) 10 (20%) 8 (15%) 

Liver (50) (51) (51) (54) 

Angiectasis 6 (12%) 6 (12%) 11 (22%) 12 (22%) 

Basophilic focus 14 (28%) 8 (16%) 6 (12%) 7 (13%) 

Clear cell focus 9 (18%) 15 (29%) 15 (29%) 18 (33%) 

Congestion 3 (6%) 

Cyst 2 (4%) 

Degeneration, cystic 15 (30%) 19 (37%) 10 (20%) 13 (24%) 

Developmental malformation 2 (4%) 

Eosinophilic focus 7 (14%) 5 (10%) 14 (27%) 12 (22%) 

Granuloma 2 (4%) 2 (4%) 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 93 

TABLE A5
 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Alimentary System (continued) 

Liver (continued) (50) (51) (51) (54) 

Hematopoietic cell proliferation 5 (10%) 1 (2%) 

Hepatodiaphragmatic nodule 1 (2%) 1 (2%) 3 (6%) 

Inflammation, subacute 9 (18%) 3 (6%) 

Mixed cell focus 1 (2%) 10 (20%) 9 (18%) 10 (19%) 

Necrosis, focal 2 (4%) 11 (22%) 3 (6%) 3 (6%) 

Thrombosis 2 (4%) 4 (8%) 1 (2%) 6 (11%) 

Bile duct, hyperplasia 49 (98%) 26 (51%) 18 (35%) 32 (59%) 

Bile duct, pigmentation 38 (75%) 51 (100%) 54 (100%) 

Centrilobular, atrophy 14 (28%) 17 (33%) 8 (16%) 15 (28%) 

Centrilobular, necrosis 1 (2%) 

Hepatocyte, cytologic alterations 20 (39%) 44 (86%) 42 (78%) 

Hepatocyte, pigmentation 22 (43%) 45 (88%) 51 (94%) 

Hepatocyte, vacuolization cytoplasmic 5 (10%) 1 (2%) 2 (4%) 

Kupffer cell, pigmentation 7 (14%) 15 (29%) 23 (45%) 26 (48%) 

Mesentery (11) (12) (9) (13) 

Accessory spleen 1 (8%) 

Angiectasis 1 (8%) 1 (8%) 

Cyst 1 (8%) 

Fat, necrosis 11 (100%) 12 (100%) 9 (100%) 12 (92%) 

Pancreas (50) (51) (51) (54) 

Atrophy 26 (52%) 30 (59%) 23 (45%) 22 (41%) 

Basophilic focus 1 (2%) 3 (6%) 2 (4%) 1 (2%) 

Edema 3 (6%) 10 (20%) 16 (31%) 22 (41%) 

Metaplasia 1 (2%) 1 (2%) 

Thrombosis 2 (4%) 

Acinar cell, cytoplasmic alteration 5 (10%) 11 (22%) 8 (15%) 

Acinar cell, hyperplasia, focal 6 (12%) 3 (6%) 6 (12%) 9 (17%) 

Salivary glands (50) (51) (50) (54) 

Atrophy 1 (2%) 1 (2%) 5 (10%) 7 (13%) 

Basophilic focus 1 (2% 

Edema 1 (2%) 6 (12%) 21 (39%) 

Inflammation, chronic 1 (2%) 

Stomach, forestomach (50) (51) (51) (54) 

Edema 5 (10%) 10 (20%) 15 (29%) 13 (24%) 

Perforation 1 (2%) 

Ulcer 5 (10%) 3 (6%) 5 (10%) 5 (9%) 

Mucosa, hyperplasia 4 (8%) 13 (25%) 19 (37%) 21 (39%) 

Stomach, glandular (50) (51) (51) (54) 

Edema 7 (14%) 2 (4%) 7 (14%) 8 (15%) 

Erosion 2 (4%) 1 (2%) 1 (2%) 

Ulcer 1 (2%) 

Tongue (1) (1) (4) 

Hyperplasia 1 (25%) 

Epithelium, hyperplasia 1 (25%) 

Tooth (1) (3) 

Necrosis 1 (100%) 2 (67%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

94 D&C Yellow No. 11, NTP TR 463 

TABLE A5
 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Cardiovascular System 

Blood vessel (50) (51) (51) (53) 

Hypertrophy 1 (2%) 3 (6%) 3 (6%) 

Inflammation, subacute 1 (2%) 2 (4%) 3 (6%) 

Mineralization 1 (2%) 

Necrosis 1 (2%) 

Thrombosis 3 (6%) 2 (4%) 3 (6%) 

Heart (50) (51) (51) (54) 

Cardiomyopathy 39 (78%) 32 (63%) 33 (65%) 27 (50%) 

Mineralization 2 (4%) 

Thrombosis 1 (2%) 

Endocrine System 

Adrenal cortex (50) (51) (51) (54) 

Accessory adrenal cortical nodule 10 (20%) 20 (39%) 12 (24%) 13 (24%) 

Angiectasis 1 (2%) 1 (2%) 

Degeneration, fatty 11 (22%) 5 (10%) 7 (14%) 6 (11%) 

Hyperplasia, diffuse 2 (4%) 

Hyperplasia, focal 1 (2%) 2 (4%) 2 (4%) 

Hypertrophy, focal 5 (10%) 1 (2%) 2 (4%) 1 (2%) 

Necrosis 1 (2%) 1 (2%) 

Adrenal medulla (50) (50) (51) (54) 

Hyperplasia 13 (26%) 20 (40%) 18 (35%) 19 (35%) 

Islets, pancreatic (50) (51) (51) (54) 

Hyperplasia 1 (2%) 

Parathyroid gland (47) (47) (48) (52) 

Cyst 1 (2%) 

Hyperplasia 3 (6%) 9 (19%) 15 (31%) 17 (33%) 

Pituitary gland (50) (50) (50) (52) 

Pars distalis, angiectasis 6 (12%) 2 (4%) 6 (12%) 3 (6%) 

Pars distalis, cyst 3 (6%) 5 (10%) 5 (10%) 7 (13%) 

Pars distalis, hyperplasia, focal 11 (22%) 7 (14%) 6 (12%) 9 (17%) 

Pars intermedia, angiectasis 1 (2%) 1 (2%) 

Pars intermedia, cyst 2 (4%) 3 (6%) 3 (6%) 

Pars nervosa, developmental malformation 1 (2%) 

Thyroid gland (50) (51) (50) (54) 

Ultimobranchial cyst 1 (2%) 4 (8%) 4 (7%) 

C-cell, hyperplasia 2 (4%) 6 (12%) 1 (2%) 6 (11%) 

Follicle, cyst 1 (2%) 2 (4%) 6 (12%) 7 (13%) 

General Body System 

None 

Genital System 

Epididymis (49) (51) (51) (54) 

Atypia cellular 19 (39%) 27 (53%) 27 (53%) 23 (43%) 

Edema 1 (2%) 3 (6%) 5 (10%) 14 (26%) 

Granuloma sperm 1 (2%) 1 (2%) 

Hypospermia 25 (51%) 37 (73%) 38 (75%) 29 (54%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 95 

TABLE A5
 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Genital System (continued) 

Preputial gland (49) (50) (51) (54) 

Cyst 3 (6%) 4 (8%) 1 (2%) 

Hyperplasia 2 (4%) 1 (2%) 2 (4%) 

Inflammation, chronic 10 (20%) 6 (12%) 10 (20%) 4 (7%) 

Inflammation, suppurative 8 (16%) 5 (10%) 7 (14%) 1 (2%) 

Prostate (50) (51) (51) (53) 

Corpora amylacea 18 (36%) 28 (55%) 24 (47%) 17 (32%) 

Edema 1 (2%) 2 (4%) 2 (4%) 12 (23%) 

Inflammation, suppurative 34 (68%) 22 (43%) 31 (61%) 34 (64%) 

Epithelium, hyperplasia 6 (12%) 5 (10%) 3 (6%) 1 (2%) 

Seminal vesicle (50) (51) (51) (54) 

Edema 1 (2%) 1 (2%) 

Testes (49) (51) (51) (54) 

Interstitial cell, hyperplasia 6 (12%) 4 (8%) 4 (8%) 3 (6%) 

Seminiferous tubule, atrophy 2 (4%) 4 (8%) 2 (4%) 5 (9%) 

Hematopoietic System 

Bone marrow (50) (51) (51) (54) 

Hemorrhage 5 (10%) 3 (6%) 

Hyperplasia 3 (6%) 1 (2%) 4 (8%) 4 (7%) 

Myelofibrosis 3 (6%) 3 (6%) 1 (2%) 4 (7%) 

Lymph node (20) (23) (26) (45) 

Hemorrhage 2 (8%) 4 (9%) 

Hyperplasia, lymphoid 2 (8%) 4 (9%) 

Pigmentation 1 (4%) 2 (4%) 

Axillary, ectasia 1 (4%) 

Axillary, hemorrhage 1 (4%) 

Axillary, hyperplasia, lymphoid 1 (4%) 

Axillary, pigmentation 1 (4%) 

Deep cervical, hemorrhage 2 (8%) 

Deep cervical, hyperplasia, lymphoid 1 (4%) 

Deep cervical, pigmentation 2 (10%) 4 (15%) 1 (2%) 

Iliac, ectasia 1 (5%) 

Iliac, hemorrhage 1 (2%) 

Iliac, hyperplasia, lymphoid 1 (2%) 

Iliac, pigmentation 1 (4%) 1 (2%) 

Inguinal, hemorrhage 2 (8%) 2 (4%) 

Inguinal, hyperplasia, lymphoid 2 (8%) 2 (4%) 

Mediastinal, ectasia 3 (12%) 1 (2%) 

Mediastinal, hemorrhage 3 (13%) 7 (27%) 16 (36%) 

Mediastinal, hyperplasia, lymphoid 9 (35%) 17 (38%) 

Mediastinal, pigmentation 8 (40%) 10 (43%) 14 (54%) 21 (47%) 

Pancreatic, ectasia 2 (10%) 

Pancreatic, hemorrhage 1 (4%) 1 (4%) 

Pancreatic, hyperplasia, lymphoid 1 (4%) 6 (13%) 

Pancreatic, pigmentation 1 (5%) 3 (13%) 3 (12%) 12 (27%) 

Renal, ectasia 1 (2%) 

Renal, hemorrhage 1 (4%) 2 (4%) 

Renal, hyperplasia, lymphoid 1 (4%) 4 (9%) 

Renal, pigmentation 3 (15%) 1 (4%) 11 (24%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

96 D&C Yellow No. 11, NTP TR 463 

TABLE A5
 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Hematopoietic System (continued) 

Lymph node, mandibular (50) (51) (50) (53) 

Ectasia 7 (14%) 8 (16%) 9 (18%) 3 (6%) 

Hemorrhage 6 (12%) 4 (8%) 8 (16%) 11 (21%) 

Hyperplasia, lymphoid 8 (16%) 12 (24%) 22 (44%) 25 (47%) 

Pigmentation 5 (10%) 5 (10%) 4 (8%) 7 (13%) 

Lymph node, mesenteric (50) (50) (51) (54) 

Ectasia 6 (12%) 1 (2%) 2 (4%) 2 (4%) 

Hemorrhage 9 (18%) 7 (13%) 

Hyperplasia, lymphoid 3 (6%) 2 (4%) 10 (20%) 14 (26%) 

Pigmentation 1 (2%) 

Spleen (50) (51) (51) (54) 

Congestion 1 (2%) 3 (6%) 1 (2%) 

Fibrosis 15 (30%) 16 (31%) 19 (37%) 17 (31%) 

Hematopoietic cell proliferation 15 (30%) 12 (24%) 16 (31%) 16 (30%) 

Metaplasia, lipocyte 2 (4%) 

Necrosis 1 (2%) 

Pigmentation 9 (18%) 12 (24%) 14 (27%) 11 (20%) 

Lymphoid follicle, hyperplasia 1 (2%) 

Thymus (48) (49) (49) (53) 

Hyperplasia 1 (2%) 

Integumentary System 

Mammary gland (48) (48) (49) (53) 

Hyperplasia 23 (48%) 11 (23%) 12 (24%) 7 (13%) 

Skin (50) (50) (51) (54) 

Cyst epithelial inclusion 3 (6%) 1 (2%) 1 (2%) 

Hyperkeratosis 4 (8%) 3 (6%) 3 (6%) 8 (15%) 

Ulcer 2 (4%) 1 (2%) 

Epidermis, hyperplasia 5 (10%) 5 (10%) 5 (10%) 9 (17%) 

Subcutaneous tissue, edema 1 (2%) 4 (8%) 16 (30%) 

Musculoskeletal System 

Bone (50) (51) (51) (54) 

Fibrous osteodystrophy 2 (4%) 8 (16%) 18 (35%) 14 (26%) 

Hyperostosis 1 (2%) 1 (2%) 

Skeletal muscle (1) (2) (5) (9) 

Edema 1 (50%) 4 (80%) 9 (100%) 

Nervous System 

Brain (50) (51) (51) (54) 

Developmental malformation 8 (16%) 4 (8%) 4 (8%) 3 (6%) 

Hemorrhage 2 (4%) 1 (2%) 

Hydrocephalus 2 (4%) 1 (2%) 1 (2%) 

Thrombosis 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 97 

TABLE A5
 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Respiratory System 

Lung (50) (51) (51) (54) 

Congestion 2 (4%) 

Cyst 1 (2%) 

Edema 1 (2%) 6 (12%) 10 (20%) 19 (35%) 

Hemorrhage 2 (4%) 2 (4%) 2 (4%) 

Infiltration cellular, histiocyte 17 (34%) 13 (25%) 18 (35%) 14 (26%) 

Inflammation, subacute 2 (4%) 4 (8%) 2 (4%) 1 (2%) 

Metaplasia, osseous 2 (4%) 

Alveolar epithelium, hyperplasia 4 (8%) 6 (12%) 2 (4%) 2 (4%) 

Nose (50) (51) (51) (54) 

Exudate 8 (16%) 7 (14%) 10 (20%) 10 (19%) 

Foreign body 2 (4%) 2 (4%) 6 (11%) 

Mucosa, hyperplasia 5 (10%) 8 (16%) 9 (18%) 8 (15%) 

Mucosa, metaplasia, squamous 7 (14%) 3 (6%) 7 (14%) 8 (15%) 

Special Senses System 

Eye (1) (1) 

Cataract 1 (100%) 

Congestion 1 (100%) 

Retina, degeneration 1 (100%) 

Urinary System 

Kidney (50) (51) (51) (54) 

Cyst 1 (2%) 6 (12%) 7 (14%) 8 (15%) 

Hydronephrosis 2 (4%) 1 (2%) 

Inflammation, suppurative 1 (2%) 5 (10%) 4 (7%) 

Mineralization 4 (8%) 2 (4%) 

Nephropathy 50 (100%) 51 (100%) 51 (100%) 54 (100%) 

Pelvis, hemorrhage 1 (2%) 

Renal tubule, hyperplasia 4 (8%) 3 (6%) 

Renal tubule, pigmentation 18 (36%) 43 (84%) 47 (92%) 54 (100%) 

Transitional epithelium, hyperplasia 11 (22%) 23 (45%) 29 (57%) 34 (63%) 

Urinary bladder (50) (51) (51) (54) 

Hemorrhage 1 (2%) 

Transitional epithelium, hyperplasia 1 (2%) 1 (2%) 1 (2%) 1 (2%) 
98 D&C Yellow No. 11, NTP TR 463 
99 

APPENDIX B 

SUMMARY OF LESIONS IN FEMALE RATS 

IN THE 2-YEAR FEED STUDY 

OF D&C YELLOW NO. 11 

TABLE B1 

TABLE B2 

TABLE B3 

TABLE B4a 

TABLE B4b 

TABLE B4c 

TABLE B4d 

TABLE B5 

Summary of the Incidence of Neoplasms in Female Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

Individual Animal Tumor Pathology of Female Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

Statistical Analysis of Primary Neoplasms in Female Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

Historical Incidence of Hepatocellular Neoplasms 

in Untreated Female F344/N Rats ................................................... 

Historical Incidence of Renal Tubule Neoplasms 

in Untreated Female F344/N Rats ................................................... 

Historical Incidence of Oral Cavity Neoplasms 

in Untreated Female F344/N Rats ................................................... 

Historical Incidence of Clitoral Gland Neoplasms 

in Untreated Female F344/N Rats ................................................... 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 

101 

106 

124 

129 

129 

130 

130 

131 
100 D&C Yellow No. 11, NTP TR 463 
 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 101 

TABLE B1 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 a 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Disposition Summary 

Animals initially in study 

12-Month interim evaluation 

Early deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

60 

10 

25 

3 

22 

60 

9 

23 

2 

26 

60 

10 

12 

1 

37 

60 

9 

25 

3 

23 

Animals examined microscopically 60 60 60 60 

12-Month Interim Evaluation 

Endocrine System 

Pituitary gland 

Pars distalis, adenoma

(10) (9) (10) (9) 

 2 (22%) 

Genital System 

Clitoral gland 

Carcinoma

Uterus 

Polyp stromal

(10)

(10)

 2 (20%) 

(9) 

 1 (11%) 

(9) 

 2 (22%) 

(10)

(10)

 3 (30%) 

(9) 

 1 (11%) 

(9) 

Hematopoietic System 

Spleen 

Fibrous histiocytoma

(10) (9) (10) (9) 

 1 (11%) 

Integumentary System 

Mammary gland 

Carcinoma

Skin 

Subcutaneous tissue, schwannoma malignant

(10)

 1 (10%) 

(10)

 1 (10%) 

(9) 

(9) 

(10)

(10)

 (9) 

 1 (11%) 

(9) 

 1 (11%) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

General Body System 

Musculoskeletal System 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

102 D&C Yellow No. 11, NTP TR 463 

TABLE B1
 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study 

Alimentary System 

Intestine large, colon (50) (50) (50) (51) 

Carcinoma 1 (2%) 1 (2%) 

Intestine large, rectum (49) (50) (49) (50) 

Histiocytic sarcoma 1 (2%) 

Intestine large, cecum (50) (51) (49) (51) 

Intestine small, jejunum (49) (51) (50) (51) 

Leiomyosarcoma 1 (2%) 

Intestine small, ileum (48) (51) (49) (49) 

Liver (50) (51) (50) (51) 

Cholangiocarcinoma 1 (2%) 1 (2%) 

Hepatocellular carcinoma 1 (2%) 

Hepatocellular adenoma 1 (2%) 5 (10%) 4 (8%) 

Hepatocellular adenoma, multiple 1 (2%) 

Osteosarcoma, metastatic, bone 1 (2%) 

Mesentery (11) (18) (8) (12) 

Cholangiocarcinoma, metastatic, liver 1 (8%) 

Osteosarcoma, metastatic, bone 1 (6%) 

Oral mucosa (1) (1) (1) 

Squamous cell carcinoma 1 (100%) 

Squamous cell papilloma 1 (100%) 1 (100%) 

Pancreas (50) (51) (50) (51) 

Osteosarcoma, metastatic, bone 1 (2%) 

Salivary glands (50) (46) (50) (50) 

Stomach, forestomach (50) (51) (50) (51) 

Squamous cell carcinoma 1 (2%) 

Squamous cell papilloma 2 (4%) 2 (4%) 

Stomach, glandular (50) (51) (50) (51) 

Tongue (1) 

Squamous cell carcinoma 1 (100%) 

Cardiovascular System 

Heart (50) (51) (50) (51) 

Osteosarcoma, metastatic, bone 1 (2%) 

Schwannoma malignant 1 (2%) 

Endocrine System 

Adrenal cortex (50) (51) (50) (51) 

Adenoma 1 (2%) 

Adrenal medulla (48) (51) (50) (51) 

Pheochromocytoma malignant 1 (2%) 1 (2%) 

Pheochromocytoma complex 1 (2%) 

Pheochromocytoma benign 2 (4%) 1 (2%) 2 (4%) 1 (2%) 

Islets, pancreatic (50) (51) (49) (51) 

Adenoma 1 (2%) 1 (2%) 1 (2%) 

Parathyroid gland (48) (48) (48) (50) 

Adenoma 1 (2%) 1 (2%) 

Pituitary gland (50) (51) (50) (51) 

Pars distalis, adenoma 23 (46%) 23 (45%) 18 (36%) 20 (39%) 

Pars distalis, adenoma, multiple 1 (2%) 

Pars distalis, carcinoma 2 (4%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 103 

TABLE B1
 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Endocrine System (continued) 

Thyroid gland (50) (51) (50) (51) 

Bilateral, follicular cell, carcinoma 1 (2%) 

C-cell, adenoma 2 (4%) 2 (4%) 4 (8%) 5 (10%) 

C-cell, adenoma, multiple 1 (2%) 

C-cell, carcinoma 1 (2%) 1 (2%) 

Follicular cell, adenoma 1 (2%) 2 (4%) 

Follicular cell, carcinoma 1 (2%) 

General Body System 

None 

Genital System 

Clitoral gland (49) (50) (49) (51) 

Adenoma 11 (22%) 4 (8%) 5 (10%) 4 (8%) 

Carcinoma 5 (10%) 2 (4%) 6 (12%) 2 (4%) 

Bilateral, carcinoma 1 (2%) 

Ovary (50) (51) (50) (51) 

Granulosa cell tumor malignant 1 (2%) 

Granulosa cell tumor benign 1 (2%) 

Osteosarcoma, metastatic, bone 1 (2%) 

Uterus (50) (51) (50) (51) 

Adenoma 1 (2%) 

Carcinoma 1 (2%) 

Cholangiocarcinoma, metastatic, liver 1 (2%) 

Polyp stromal 11 (22%) 11 (22%) 7 (14%) 6 (12%) 

Polyp stromal, multiple 1 (2%) 

Schwannoma malignant 1 (2%) 1 (2%) 

Hematopoietic System 

Bone marrow (50) (51) (50) (51) 

Lymph node (9) (11) (11) (15) 

Lymph node, mandibular (50) (51) (50) (49) 

Lymph node, mesenteric (50) (51) (50) (51) 

Lymph node, mediastinal (1) (1) 

Cholangiocarcinoma, metastatic, liver 1 (100%) 

Spleen (50) (50) (50) (51) 

Thymus (50) (49) (49) (49) 

Integumentary System 

Mammary gland (50) (51) (50) (51) 

Adenoma 2 (4%) 

Carcinoma 4 (8%) 5 (10%) 2 (4%) 3 (6%) 

Carcinoma, multiple 1 (2%) 

Fibroadenoma 14 (28%) 13 (25%) 18 (36%) 17 (33%) 

Fibroadenoma, multiple 7 (14%) 9 (18%) 5 (10%) 9 (18%) 

Histiocytic sarcoma 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

104 D&C Yellow No. 11, NTP TR 463 

TABLE B1
 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Integumentary System (continued) 

Skin 

Basal cell carcinoma

Histiocytic sarcoma

Squamous cell carcinoma

Squamous cell papilloma

Subcutaneous tissue, fibroma

(49) 

 1 (2%) 

(51) 

 1 (2%) 

(50) 

 2 (4%) 

(51) 

 1 (2%) 

 1 (2%) 

 1 (2%) 

Musculoskeletal System 

Bone 

Osteosarcoma

Skeletal muscle

Osteosarcoma, metastatic, bone

Rhabdomyosarcoma

(50) 

 1 (2%) 

(51) 

 1 (2%) 

(2)

 1 (50%) 

(50) (51) 

 1 (2%) 

(1) 

 1 (100%) 

Nervous System 

Brain 

Astrocytoma malignant

Carcinoma, metastatic, pituitary gland

(50) (51) 

 1 (2%) 

(50) (51) 

 2 (4%) 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar adenoma, multiple

Alveolar/bronchiolar carcinoma

Carcinoma, metastatic, uterus

Cholangiocarcinoma, metastatic, liver

Osteosarcoma, metastatic, bone

(50) 

 1 (2%) 

 1 (2%) 

(51) 

 1 (2%) 

(50) 

 2 (4%) 

(51) 

 1 (2%) 

 2 (4%) 

 1 (2%) 

 1 (2%) 

Special Senses System 

Zymbal’s gland

Carcinoma

 (1) 

 1 (100%) 

Urinary System 

Kidney 

Sarcoma

Renal tubule, carcinoma

Transitional epithelium, carcinoma

Transitional epithelium, hemangioma

Urinary bladder 

Papilloma

(50) 

 1 (2%) 

(50) 

(51) 

(51) 

(50) 

 1 (2%) 

 1 (2%) 

 1 (2%) 

(50) 

(51) 

(51) 

 1 (2%) 

Systemic Lesions 

Multiple organsb 

Histiocytic sarcoma

Leukemia mononuclear

Lymphoma malignant

(50) 

 16 (32%) 

(51) 

 21 (41%) 

 1 (2%) 

(50) 

 19 (38%) 

(51) 

 1 (2%) 

 16 (31%) 
105 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE B1
 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Neoplasm Summary 

Total animals with primary neoplasmsc 

12-Month interim evaluation 4 3 3 4 

2-Year study 48 50 50 49 

Total primary neoplasms 

12-Month interim evaluation 4 3 3 6 

2-Year study 111 108 107 113 

Total animals with benign neoplasms 

12-Month interim evaluation 2 2 3 2 

2-Year study 42 44 41 41 

Total benign neoplasms 

12-Month interim evaluation 2 2 3 2 

2-Year study 78 72 71 76 

Total animals with malignant neoplasms 

12-Month interim evaluation 2 1 2 

2-Year study 30 29 30 28 

Total malignant neoplasms 

12-Month interim evaluation 2 1 4 

2-Year study 33 36 36 37 

Total animals with metastatic neoplasms 

2-Year study 1 1 4 

Total metastatic neoplasms 

2-Year study 1 5 8 

a Number of animals examined microscopically at the site and the number of animals with neoplasm b Number of animals with any tissue examined microscopically
 

Primary neoplasms: all neoplasms except metastatic neoplasms
 
 

 

 

 

 

 

 

 

 

 

 

106 D&C Yellow No. 11, NTP TR 463 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm 

4 5 5 5 5 5 5 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 

Number of Days on Study 8 3 7 7 7 8 8 0 0 2 5 5 5 6 7 8 9 0 0 1 1 2 2 3 3 

1 7 0 2 2 9 9 2 7 9 0 1 6 0 1 4 9 0 1 2 9 2 9 0 0 

2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 

Carcass ID Number 9 9 8 6 7 4 8 6 5 9 4 8 5 5 4 5 6 9 4 5 4 4 5 5 7 

1 6 6 3 1 8 8 5 7 8 9 1 2 4 5 8 2 4 4 5 7 6 0 1 3 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + M + + + + + 

Leiomyosarcoma X 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + M + + + + + 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Cholangiocarcinoma X 

Mesentery + + + + + + 

Oral mucosa 

Squamous cell papilloma 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Tooth + 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adrenal medulla + + + + + + + + M + + + + + + + + + + + + + + + + 

Pheochromocytoma complex X 

Pheochromocytoma benign X 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X 

Parathyroid gland + + + + + + + + + + + + + + + + + + + + + + M + + 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X X X X X X X X X X X X 

Pars distalis, adenoma, multiple X 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

C-cell, adenoma 

General Body System 

None 

Genital System 

Clitoral gland + + + + + + + + + + + + + + + + + + M + + + + + + 

Adenoma X X X 

Carcinoma X X 

Bilateral, carcinoma 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + 

+: Tissue examined microscopically M: Missing tissue X: Lesion present 

A: Autolysis precludes examination I: Insufficient tissue Blank: Not examined 
 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 107 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 3 3 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

4 4 4 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 

2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 3 Total 

Carcass ID Number 7 8 9 4 4 5 5 6 6 6 6 6 7 7 7 7 7 8 8 8 8 9 9 9 0 Tissues/ 

9 4 3 1 3 6 9 1 4 6 8 9 0 4 5 6 7 0 2 5 7 0 5 7 0 Tumors 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + M + + + + + 49 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Leiomyosarcoma 1 

Intestine small, ileum + + + + + + + + I + + + + + + + + + + + + + + + + 48 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Cholangiocarcinoma 1 

Mesentery + + + + + 11 

Oral mucosa + 1 

Squamous cell papilloma X 1 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Tooth 1 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adrenal medulla M + + + + + + + + + + + + + + + + + + + + + + + + 48 

Pheochromocytoma complex 1 

Pheochromocytoma benign X 2 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adenoma 1 

Parathyroid gland + + + + + + + + + + + + + + + + + + + + + + M + + 48 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pars distalis, adenoma X X X X X X X X X X 23 

Pars distalis, adenoma, multiple 1 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

C-cell, adenoma X X 2 

General Body System 

None 

Genital System 

Clitoral gland + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Adenoma X X X X X X X X 11 

Carcinoma X X X 5 

Bilateral, carcinoma X 1 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + 50 
 

 

 

 

 

 

 

 

 

 

 

 

 

108 D&C Yellow No. 11, NTP TR 463 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

4 5 5 5 5 5 5 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 

Number of Days on Study 8 3 7 7 7 8 8 0 0 2 5 5 5 6 7 8 9 0 0 1 1 2 2 3 3 

1 7 0 2 2 9 9 2 7 9 0 1 6 0 1 4 9 0 1 2 9 2 9 0 0 

2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 

Carcass ID Number 9 9 8 6 7 4 8 6 5 9 4 8 5 5 4 5 6 9 4 5 4 4 5 5 7 

1 6 6 3 1 8 8 5 7 8 9 1 2 4 5 8 2 4 4 5 7 6 0 1 3 

Genital System (continued) 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Polyp stromal X X X X X X X X 

Polyp stromal, multiple 

Schwannoma malignant X 

Vagina M M M M M 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node + + + + + +
 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + +
 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + +
 

Thymus + + + + + + + + + + + + + + + + + + + + + + + + +
 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X X 

Carcinoma X X X X 

Fibroadenoma X X X X 

Fibroadenoma, multiple X X X X X 

Skin + I + + + + + + + + + + + + + + + + + + + + + + + 

Subcutaneous tissue, fibroma X 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma X 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + +
 

Peripheral nerve + +
 

Spinal cord + +
 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma 

Carcinoma, metastatic, uterus 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 

Special Senses System 

Eye 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Transitional epithelium, carcinoma X 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Leukemia mononuclear X X X X X X X X 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 109 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 0 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 3 3 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

4 4 4 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 

2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 3 Total 

Carcass ID Number 7 8 9 4 4 5 5 6 6 6 6 6 7 7 7 7 7 8 8 8 8 9 9 9 0 Tissues/ 

9 4 3 1 3 6 9 1 4 6 8 9 0 4 5 6 7 0 2 5 7 0 5 7 0 Tumors 

Genital System (continued) 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma X 1 

Polyp stromal X X X 11 

Polyp stromal, multiple X 1 

Schwannoma malignant 1 

Vagina M + 1 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node + + + 9 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Thymus + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adenoma 2 

Carcinoma 4 

Fibroadenoma X X X X X X X X X X 14 

Fibroadenoma, multiple X X 7 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Subcutaneous tissue, fibroma 1 

Musculoskeletal System 

Bone 

Osteosarcoma

+ + + + + + + + + + + + + + + + + + + + + + + + + 50 

1 

Nervous System 

Brain 

Peripheral nerve

Spinal cord 

+ + + + + + + + + + + + + + 

+

+ + + + + + + + + + + 50 

2 

3 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Alveolar/bronchiolar adenoma X 1 

Carcinoma, metastatic, uterus X 1 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Special Senses System 

Eye +

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Transitional epithelium, carcinoma 1 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Leukemia mononuclear X X X X X X X X 16 

 1 
 

 

 

 

 

 

110 D&C Yellow No. 11, NTP TR 463 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm 

3 4 5 5 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 

Number of Days on Study 0 6 4 4 0 0 2 2 2 2 3 3 4 4 5 7 8 8 9 9 0 1 1 2 3 

8 2 6 8 0 1 2 2 8 9 1 1 2 5 6 2 8 8 3 5 5 2 6 6 4 

3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 

Carcass ID Number 2 5 5 1 5 3 0 1 2 2 1 4 3 2 4 1 1 1 4 4 2 4 4 2 0 

2 9 8 5 4 6 3 3 6 0 4 0 5 4 9 0 6 8 6 1 9 4 3 1 1 

Alimentary System 

Esophagus + + + + + + + + + + + + + M + + + + + + + + + + + 

Intestine large, colon + + + M + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + I 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple X 

Osteosarcoma, metastatic, bone X 

Mesentery + + + + + + + + + + + + + 

Osteosarcoma, metastatic, bone X 

Oral mucosa + 

Squamous cell papilloma X 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma, metastatic, bone X 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Squamous cell carcinoma 

Squamous cell papilloma 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 

Pheochromocytoma benign X 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma 

Parathyroid gland + + + + M + + + + + + + + + + + + + + + + + + + + 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X X X X X X X X 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

C-cell, adenoma 

C-cell, carcinoma 

Follicular cell, carcinoma 

General Body System 

None 

Genital System 

Clitoral gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma 

Carcinoma X X 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma, metastatic, bone X 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 111 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm (continued)
 

Number of Days on Study 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

0 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

7 

4 

1 

Carcass ID Number 

3 

0 

2 

3 

0 

4 

3 

0 

6 

3 

0 

8 

3 

0 

9 

3 

1 

1 

3 

1 

7 

3 

2 

3 

3 

2 

5 

3 

2 

8 

3 

3 

0 

3 

3 

1 

3 

3 

3 

3 

3 

4 

3 

3 

8 

3 

3 

9 

3 

4 

2 

3 

4 

5 

3 

4 

7 

3 

5 

0 

3 

5 

1 

3 

5 

2 

3 

5 

3 

3 

5 

5 

3 

5 

6 

3 

6 

0 

Total 

Tissues/ 

Tumors 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + M I + + + + 48 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma X 1 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Hepatocellular adenoma X 1 

Hepatocellular adenoma, multiple 1 

Osteosarcoma, metastatic, bone 1 

Mesentery + + + + + 18 

Osteosarcoma, metastatic, bone 1 

Oral mucosa 1 

Squamous cell papilloma 1 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Osteosarcoma, metastatic, bone 1 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + 46 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Squamous cell carcinoma X 1 

Squamous cell papilloma X X 2 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adenoma X 1 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Pheochromocytoma benign 1 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adenoma X 1 

Parathyroid gland + + + M + + + + + + M + + + + + + + + + + + + + + + 48 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Pars distalis, adenoma X X X X X X X X X X X X X X 23 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

C-cell, adenoma X X 2 

C-cell, carcinoma X 1 

Follicular cell, carcinoma X 1 

General Body System 

None 

Genital System 

Clitoral gland + + + + + + + + + + + + + + + + + + + + M + + + + + 50 

Adenoma X X X X 4 

Carcinoma 2 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Osteosarcoma, metastatic, bone 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

112 D&C Yellow No. 11, NTP TR 463 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm (continued)
 

3 4 5 5 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 7 7 7 7 7 

Number of Days on Study 0 6 4 4 0 0 2 2 2 2 3 3 4 4 5 7 8 8 9 9 0 1 1 2 3 

8 2 6 8 0 1 2 2 8 9 1 1 2 5 6 2 8 8 3 5 5 2 6 6 4 

3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 

Carcass ID Number 2 5 5 1 5 3 0 1 2 2 1 4 3 2 4 1 1 1 4 4 2 4 4 2 0 

2 9 8 5 4 6 3 3 6 0 4 0 5 4 9 0 6 8 6 1 9 4 3 1 1 

Genital System (continued) 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + 

Polyp stromal X X X X X X X 

Schwannoma malignant X 

Vagina M M M M M 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node + + + + + + + + +
 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mediastinal +
 

Spleen + + + + + + + + + + + + + + + + + + + + M + + + +
 

Thymus + + + M + + + + + + + + + + + + + + + + M + + + +
 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma X X 

Fibroadenoma X X X X X X X 

Fibroadenoma, multiple X X X X X 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 

Squamous cell papilloma 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma X 

Skeletal muscle + 

Osteosarcoma, metastatic, bone X 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 

Astrocytoma malignant X 

Peripheral nerve + + 

Spinal cord + + 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma, multiple 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 

Special Senses System 

Ear + +
 

Eye +
 

Harderian gland
 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Leukemia mononuclear X X X X X X X X X X X X X X 

Lymphoma malignant X 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 113 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 500 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 

3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 3 Total 

Carcass ID Number 0 0 0 0 0 1 1 2 2 2 3 3 3 3 3 3 4 4 4 5 5 5 5 5 5 6 Tissues/ 

2 4 6 8 9 1 7 3 5 8 0 1 3 4 8 9 2 5 7 0 1 2 3 5 6 0 Tumors 

Genital System (continued) 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Polyp stromal X X X X 11 

Schwannoma malignant 1 

Vagina 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node + + 11 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node, mediastinal 1 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Thymus + + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Carcinoma X X X 5 

Fibroadenoma X X X X X X 13 

Fibroadenoma, multiple X X X X 9 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Squamous cell papilloma X 1 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Osteosarcoma 1 

Skeletal muscle + 2 

Osteosarcoma, metastatic, bone 1 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Astrocytoma malignant 1 

Peripheral nerve 2 

Spinal cord 2 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Alveolar/bronchiolar adenoma, multiple X 1 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Special Senses System 

Ear 2 

Eye + 2 

Harderian gland + 1 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Leukemia mononuclear X X X X X X X 21 

Lymphoma malignant 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

114 D&C Yellow No. 11, NTP TR 463 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm 

4 5 5 5 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 5 3 8 9 1 3 5 6 7 0 1 2 2 4 4 4 4 4 4 4 4 4 4 4 4 

6 2 8 7 0 8 8 0 1 1 9 2 2 0 0 0 0 0 0 0 0 0 0 0 0 

4 3 4 3 4 4 3 3 4 3 3 3 4 3 3 3 3 3 3 3 3 3 3 3 3 

Carcass ID Number 1 7 0 7 1 0 7 7 0 6 9 9 1 6 6 6 6 6 6 7 7 7 7 8 8 

4 8 9 0 5 3 2 6 1 3 6 7 7 1 2 5 7 8 9 4 5 7 9 0 2 

Alimentary System 

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Intestine large, rectum + + + + + + + + M + + + + + + + + + + + + + + + + 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + M + + + + 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, ileum + + + + + + + + + + + M + + + + + + + + + + + + + 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Hepatocellular adenoma X X 

Mesentery + + + + + 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 

Pheochromocytoma malignant X 

Pheochromocytoma benign X X 

Islets, pancreatic + I + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X 

Parathyroid gland + + M + M + + + + + + + + + + + + + + + + + + + + 

Adenoma 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X X X X X X X 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

C-cell, adenoma X X 

C-cell, carcinoma X 

Follicular cell, adenoma X 

General Body System 

None 

Genital System 

Clitoral gland + + M + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X 

Carcinoma X X X 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + 

Granulosa cell tumor malignant X 

Granulosa cell tumor benign 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + 

Polyp stromal X X X 

Vagina M 
 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 115 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

3 3 3 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 4 4 Total 

Carcass ID Number 8 8 8 8 8 8 9 9 9 9 9 9 9 0 0 0 0 0 0 1 1 1 1 1 2 Tissues/ 

3 4 5 6 7 9 0 2 3 4 5 8 9 0 2 4 5 6 7 0 1 3 6 9 0 Tumors 

Alimentary System 

Esophagus + + + + + + + + + + + + M + + + + + + + + + + + + 49 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma X 1 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hepatocellular adenoma X X X 5 

Mesentery + + + 8 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pheochromocytoma malignant 1 

Pheochromocytoma benign 2 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Adenoma 1 

Parathyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 48 

Adenoma X 1 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pars distalis, adenoma X X X X X X X X X X 18 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

C-cell, adenoma X X 4 

C-cell, carcinoma 1 

Follicular cell, adenoma 1 

General Body System 

None 

Genital System 

Clitoral gland + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Adenoma X X X X 5 

Carcinoma X X X 6 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Granulosa cell tumor malignant 1 

Granulosa cell tumor benign X 1 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Polyp stromal X X X X 7 

Vagina 
 

 

 

 

 

 

 

 

116 D&C Yellow No. 11, NTP TR 463 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

4 5 5 5 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study 5 3 8 9 1 3 5 6 7 0 1 2 2 4 4 4 4 4 4 4 4 4 4 4 4 

6 2 8 7 0 8 8 0 1 1 9 2 2 0 0 0 0 0 0 0 0 0 0 0 0 

4 3 4 3 4 4 3 3 4 3 3 3 4 3 3 3 3 3 3 3 3 3 3 3 3 

Carcass ID Number 1 7 0 7 1 0 7 7 0 6 9 9 1 6 6 6 6 6 6 7 7 7 7 8 8 

4 8 9 0 5 3 2 6 1 3 6 7 7 1 2 5 7 8 9 4 5 7 9 0 2 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node + + + + + + + + +
 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + +
 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + +
 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + +
 

Thymus + + + + M + + + + + + + + + + + + + + + + + + + +
 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma 

Carcinoma, multiple X 

Fibroadenoma X X X X X X X X X 

Fibroadenoma, multiple X X X X 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 

Squamous cell carcinoma X X 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma X 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 

Special Senses System 

None 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Sarcoma 

Renal tubule, carcinoma 

Transitional epithelium, hemangioma X 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Leukemia mononuclear X X X X X X X X X X X X 
 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 117 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 1,700 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study	 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

3 3 3 3 3 3 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 4 4 Total 

Carcass ID Number 8 8 8 8 8 8 9 9 9 9 9 9 9 0 0 0 0 0 0 1 1 1 1 1 2 Tissues/ 

3 4 5 6 7 9 0 2 3 4 5 8 9 0 2 4 5 6 7 0 1 3 6 9 0 Tumors 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node + + 11 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Thymus + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma X X 2 

Carcinoma, multiple 1 

Fibroadenoma X X X X X X X X X 18 

Fibroadenoma, multiple X 5 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Squamous cell carcinoma 2 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Respiratory System 

Lung	 + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Alveolar/bronchiolar adenoma X 2 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Special Senses System 

None 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Sarcoma X 1 

Renal tubule, carcinoma X 1 

Transitional epithelium, hemangioma 1 

Urinary bladder	 + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Leukemia mononuclear X X X X X X X 19 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

118 D&C Yellow No. 11, NTP TR 463 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm 

1 4 4 4 5 5 5 5 5 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 

Number of Days on Study 4 4 4 6 0 3 6 9 9 4 5 5 5 5 7 9 0 1 2 2 2 2 2 2 2 

7 1 8 9 6 0 8 0 6 8 6 6 8 8 2 5 0 2 0 3 3 8 8 8 9 

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

Carcass ID Number 7 4 8 3 5 5 6 7 4 2 2 7 4 5 5 2 6 6 4 2 2 3 4 4 7 

3 2 0 7 6 9 2 1 6 2 1 7 4 3 0 4 1 5 3 5 8 6 0 9 0 

Alimentary System 

Esophagus + + + + + + M + + + + + + + + + + + + + + + + + M 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + I + + + 

Histiocytic sarcoma X 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + 

Intestine small, ileum + + + + + + + + + + + + + M + + M + + + + + + + + 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 

Cholangiocarcinoma X 

Hepatocellular carcinoma X 

Hepatocellular adenoma X 

Mesentery + + + + + + 

Cholangiocarcinoma, metastatic, liver X 

Oral mucosa 

Squamous cell carcinoma 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + 

Squamous cell papilloma X 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + 

Tongue 

Squamous cell carcinoma 

Cardiovascular System 

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma, metastatic, bone X 

Schwannoma malignant 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 

Pheochromocytoma malignant X 

Pheochromocytoma benign 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 

Parathyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Pars distalis, adenoma X X X X X X X X X X 

Pars distalis, carcinoma X 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Bilateral, follicular cell, carcinoma 

C-cell, adenoma X X 

C-cell, adenoma, multiple X 

Follicular cell, adenoma X X 

General Body System 

None 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 119 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study	 2 3 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

9 3 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 Total 

Carcass ID Number 7 3 3 2 2 3 3 3 3 3 4 4 5 5 5 5 5 6 6 6 6 6 7 7 7 7 Tissues/ 

6 2 3 6 9 0 1 5 8 9 5 8 1 4 5 7 8 3 4 6 7 8 4 5 8 9 Tumors 

Alimentary System 

Esophagus + + + + + M + + + + + M + + + + + + + + + + + + + + 47 

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Histiocytic sarcoma 1 

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, duodenum + + + + + + + + + + + + + + + M + + + + + + + + + + 50 

Intestine small, jejunum + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Cholangiocarcinoma 1 

Hepatocellular carcinoma 1 

Hepatocellular adenoma X X X 4 

Mesentery + + + + + + 12 

Cholangiocarcinoma, metastatic, liver 1 

Oral mucosa + 1 

Squamous cell carcinoma X 1 

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Squamous cell papilloma X 2 

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Tongue + 1 

Squamous cell carcinoma X 1 

Cardiovascular System 

Blood vessel	 + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Osteosarcoma, metastatic, bone 1 

Schwannoma malignant X 1 

Endocrine System 

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Pheochromocytoma malignant 1 

Pheochromocytoma benign X 1 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Parathyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + M 50 

Adenoma 1 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Pars distalis, adenoma X X X X X X X X X X 20 

Pars distalis, carcinoma X 2 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Bilateral, follicular cell, carcinoma X 1 

C-cell, adenoma X X X 5 

C-cell, adenoma, multiple 1 

Follicular cell, adenoma 2 

General Body System 

None 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

120 D&C Yellow No. 11, NTP TR 463 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

1 4 4 4 5 5 5 5 5 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 

Number of Days on Study 4 4 4 6 0 3 6 9 9 4 5 5 5 5 7 9 0 1 2 2 2 2 2 2 2 

7 1 8 9 6 0 8 0 6 8 6 6 8 8 2 5 0 2 0 3 3 8 8 8 9 

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

Carcass ID Number 7 4 8 3 5 5 6 7 4 2 2 7 4 5 5 2 6 6 4 2 2 3 4 4 7 

3 2 0 7 6 9 2 1 6 2 1 7 4 3 0 4 1 5 3 5 8 6 0 9 0 

Genital System 

Clitoral gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X X 

Carcinoma X 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + 

Adenoma X 

Cholangiocarcinoma, metastatic, liver X 

Polyp stromal X X X 

Vagina M M M M 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + 

Lymph node + + + + + + + + + + + + 

Lymph node, mandibular + + + + + + + + + + + + + + + + + + + + + + + + + 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + 

Lymph node, mediastinal + 

Cholangiocarcinoma, metastatic, liver X 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + 

Thymus + + + + M + + + + + + + + + + + + + + + + + + + + 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma X X 

Fibroadenoma X X X X X X 

Fibroadenoma, multiple X X X X X 

Histiocytic sarcoma X 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 

Basal cell carcinoma 

Histiocytic sarcoma X 

Squamous cell papilloma 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 

Osteosarcoma X 

Skeletal muscle + 

Rhabdomyosarcoma X 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + 

Carcinoma, metastatic, pituitary gland X 

Peripheral nerve + + 

Spinal cord + + 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 121 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study	 2 3 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

9 3 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 Total 

Carcass ID Number 7 3 3 2 2 3 3 3 3 3 4 4 5 5 5 5 5 6 6 6 6 6 7 7 7 7 Tissues/ 

6 2 3 6 9 0 1 5 8 9 5 8 1 4 5 7 8 3 4 6 7 8 4 5 8 9 Tumors 

Genital System 

Clitoral gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adenoma X X 4 

Carcinoma X 2 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Adenoma 1 

Cholangiocarcinoma, metastatic, liver 1 

Polyp stromal X X X 6 

Vagina M 

Hematopoietic System 

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node + + + 15 

Lymph node, mandibular + M + + + + + + + + + + + + + + + + + + + I + + + + 49 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Lymph node, mediastinal 1 

Cholangiocarcinoma, metastatic, liver 1 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Thymus + + + + + + + + + + + + + + + M + + + + + + + + + + 49 

Integumentary System 

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Carcinoma X 3 

Fibroadenoma X X X X X X X X X X X 17 

Fibroadenoma, multiple X X X X 9 

Histiocytic sarcoma 1 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Basal cell carcinoma X 1 

Histiocytic sarcoma 1 

Squamous cell papilloma X 1 

Musculoskeletal System 

Bone + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Osteosarcoma 1 

Skeletal muscle 1 

Rhabdomyosarcoma 1 

Nervous System 

Brain + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Carcinoma, metastatic, pituitary gland X 2 

Peripheral nerve 2 

Spinal cord 2 
 

 

 

 

 

 

 

122 D&C Yellow No. 11, NTP TR 463 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

1 4 4 4 5 5 5 5 5 6 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 

Number of Days on Study 4 4 4 6 0 3 6 9 9 4 5 5 5 5 7 9 0 1 2 2 2 2 2 2 2 

7 1 8 9 6 0 8 0 6 8 6 6 8 8 2 5 0 2 0 3 3 8 8 8 9 

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

Carcass ID Number 7 4 8 3 5 5 6 7 4 2 2 7 4 5 5 2 6 6 4 2 2 3 4 4 7 

3 2 0 7 6 9 2 1 6 2 1 7 4 3 0 4 1 5 3 5 8 6 0 9 0 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma X X 

Cholangiocarcinoma, metastatic, liver X 

Osteosarcoma, metastatic, bone X 

Nose + + + + + + + + + + + + + + + + + + + + + + + + + 

Trachea + + + + + + + + + + + + + + + + + + + + + + + + + 

Special Senses System 

Ear +
 

Lacrimal gland +
 

Zymbal’s gland
 

Carcinoma 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + 

Papilloma 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 

Histiocytic sarcoma X 

Leukemia mononuclear X X X X X X X X X X X X 
 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 123 

TABLE B2
 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of D&C Yellow No. 11: 5,000 ppm (continued)
 

7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 

Number of Days on Study	 2 3 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 

9 3 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 

4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 Total 

Carcass ID Number 7 3 3 2 2 3 3 3 3 3 4 4 5 5 5 5 5 6 6 6 6 6 7 7 7 7 Tissues/ 

6 2 3 6 9 0 1 5 8 9 5 8 1 4 5 7 8 3 4 6 7 8 4 5 8 9 Tumors 

Respiratory System 

Lung + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Alveolar/bronchiolar adenoma X 1 

Alveolar/bronchiolar carcinoma 2 

Cholangiocarcinoma, metastatic, liver 1 

Osteosarcoma, metastatic, bone 1 

Nose	 + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Trachea	 + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Special Senses System 

Ear 1 

Lacrimal gland 1 

Zymbal’s gland + 1 

Carcinoma X 1 

Urinary System 

Kidney + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Urinary bladder + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Papilloma X 1 

Systemic Lesions 

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + + 51 

Histiocytic sarcoma 1 

Leukemia mononuclear X X X X 16 
124 D&C Yellow No. 11, NTP TR 463 

TABLE B3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Adrenal Medulla: Benign, Complex, or Malignant Pheochromocytoma a Overall rate 3/48 (6%) 1/51 (2%) 3/50 (6%) 2/51 (4%) b Adjusted rate 10.7% 2.4% 7.2% 7.3% c Terminal rate 1/22 (5%) 0/26 (0%) 1/37 (3%) 1/23 (4%) 

First incidence (days) 712 631 658 723 

d Life table test P=0.557N P=0.300N P=0.510N P=0.463N 

d Logistic regression test P=0.563N P=0.287N P=0.639N P=0.471N 

d Cochran-Armitage test P=0.556N 

d Fisher exact test P=0.286N P=0.641N P=0.471N 

Clitoral Gland: Adenoma 

Overall rate 11/49 (22%) 4/50 (8%) 5/49 (10%) 4/51 (8%) 

Adjusted rate 40.2% 16.0% 13.5% 13.7% 

Terminal rate 7/22 (32%) 4/25 (16%) 5/37 (14%) 2/23 (9%) 

First incidence (days) 629 740 (T) 740 (T) 656 

Life table test P=0.100N P=0.029N P=0.009N P=0.043N 

Logistic regression test P=0.086N P=0.043N P=0.035N P=0.037N 

Cochran-Armitage test P=0.098N 

Fisher exact test P=0.041N P=0.085N P=0.038N 

Clitoral Gland: Carcinoma 

Overall rate 6/49 (12%) 2/50 (4%) 6/49 (12%) 2/51 (4%) 

Adjusted rate 23.3% 5.3% 15.2% 7.5% 

Terminal rate 4/22 (18%) 0/25 (0%) 5/37 (14%) 1/23 (4%) 

First incidence (days) 660 548 456 728 

Life table test P=0.213N P=0.123N P=0.317N P=0.129N 

Logistic regression test P=0.203N P=0.131N P=0.603N P=0.116N 

Cochran-Armitage test P=0.199N 

Fisher exact test P=0.128N P=0.620N P=0.122N 

Clitoral Gland: Adenoma or Carcinoma 

Overall rate 17/49 (35%) 6/50 (12%) 11/49 (22%) 6/51 (12%) 

Adjusted rate 58.9% 20.5% 28.5% 20.6% 

Terminal rate 11/22 (50%) 4/25 (16%) 10/37 (27%) 3/23 (13%) 

First incidence (days) 629 548 456 656 

Life table test P=0.043N P=0.005N P=0.007N P=0.008N 

Logistic regression test P=0.037N P=0.007N P=0.031N P=0.005N 

Cochran-Armitage test P=0.038N 

Fisher exact test P=0.007N P=0.132N P=0.006N 

Liver: Hepatocellular Adenoma 

Overall rate 0/50 (0%) 2/51 (4%) 5/50 (10%) 4/51 (8%) 

Adjusted rate 0.0% 6.4% 13.5% 15.7% 

Terminal rate 0/22 (0%) 1/26 (4%) 5/37 (14%) 3/23 (13%) e First incidence (days) — 645 740 (T) 720 

Life table test P=0.082 P=0.252 P=0.095 P=0.072 

Logistic regression test P=0.100 P=0.241 P=0.095 P=0.068 

Cochran-Armitage test P=0.104 

Fisher exact test P=0.252 P=0.028 P=0.061 
D&C Yellow No. 11, NTP TR 463 125 

TABLE B3
 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Liver: Hepatocellular Adenoma or Carcinoma 

Overall rate 0/50 (0%) 2/51 (4%) 5/50 (10%) 5/51 (10%) 

Adjusted rate 0.0% 6.4% 13.5% 18.5% 

Terminal rate 0/22 (0%) 1/26 (4%) 5/37 (14%) 3/23 (13%) 

First incidence (days) — 645 740 (T) 720 

Life table test P=0.033 P=0.252 P=0.095 P=0.041 

Logistic regression test P=0.042 P=0.241 P=0.095 P=0.036 

Cochran-Armitage test P=0.045 

Fisher exact test P=0.252 P=0.028 P=0.030 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma 

Overall rate 1/50 (2%) 1/51 (2%) 2/50 (4%) 3/51 (6%) 

Adjusted rate 4.5% 3.8% 5.4% 10.0% 

Terminal rate 1/22 (5%) 1/26 (4%) 2/37 (5%) 1/23 (4%) 

First incidence (days) 740 (T) 740 (T) 740 (T) 590 

Life table test P=0.155 P=0.725N P=0.678 P=0.314 

Logistic regression test P=0.176 P=0.725N P=0.678 P=0.307 

Cochran-Armitage test P=0.183 

Fisher exact test P=0.748N P=0.500 P=0.316 

Mammary Gland: Fibroadenoma 

Overall rate 21/50 (42%) 22/51 (43%) 23/50 (46%) 26/51 (51%) 

Adjusted rate 62.0% 55.8% 51.9% 69.5% 

Terminal rate 10/22 (45%) 10/26 (38%) 16/37 (43%) 12/23 (52%) 

First incidence (days) 589 548 456 568 

Life table test P=0.189 P=0.510N P=0.162N P=0.292 

Logistic regression test P=0.161 P=0.516 P=0.526 P=0.207 

Cochran-Armitage test P=0.193 

Fisher exact test P=0.534 P=0.420 P=0.240 

Mammary Gland: Fibroadenoma or Adenoma 

Overall rate 22/50 (44%) 22/51 (43%) 23/50 (46%) 26/51 (51%) 

Adjusted rate 63.3% 55.8% 51.9% 69.5% 

Terminal rate 10/22 (45%) 10/26 (38%) 16/37 (43%) 12/23 (52%) 

First incidence (days) 589 548 456 568 

Life table test P=0.224 P=0.444N P=0.121N P=0.356 

Logistic regression test P=0.196 P=0.565N P=0.554N P=0.272 

Cochran-Armitage test P=0.231 

Fisher exact test P=0.545N P=0.500 P=0.308 

Mammary Gland: Carcinoma 

Overall rate 4/50 (8%) 5/51 (10%) 3/50 (6%) 3/51 (6%) 

Adjusted rate 10.6% 15.7% 7.8% 8.1% 

Terminal rate 0/22 (0%) 3/26 (12%) 2/37 (5%) 0/23 (0%) 

First incidence (days) 481 628 722 506 

Life table test P=0.356N P=0.530 P=0.359N P=0.505N 

Logistic regression test P=0.316N P=0.536 P=0.568N P=0.407N 

Cochran-Armitage test P=0.346N 

Fisher exact test P=0.513 P=0.500N P=0.489N 
126 D&C Yellow No. 11, NTP TR 463 

TABLE B3
 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Mammary Gland: Adenoma or Carcinoma 

Overall rate 6/50 (12%) 5/51 (10%) 3/50 (6%) 3/51 (6%) 

Adjusted rate 16.1% 15.7% 7.8% 8.1% 

Terminal rate 0/22 (0%) 3/26 (12%) 2/37 (5%) 0/23 (0%) 

First incidence (days) 481 628 722 506 

Life table test P=0.204N P=0.479N P=0.142N P=0.248N 

Logistic regression test P=0.181N P=0.469N P=0.290N P=0.184N 

Cochran-Armitage test P=0.199N 

Fisher exact test P=0.486N P=0.243N P=0.234N 

Mammary Gland: Fibroadenoma, Adenoma, or Carcinoma 

Overall rate 25/50 (50%) 27/51 (53%) 25/50 (50%) 28/51 (55%) 

Adjusted rate 65.9% 65.7% 55.4% 70.9% 

Terminal rate 10/22 (45%) 13/26 (50%) 17/37 (46%) 12/23 (52%) 

First incidence (days) 481 548 456 506 

Life table test P=0.354 P=0.557N P=0.087N P=0.424 

Logistic regression test P=0.338 P=0.447 P=0.534N P=0.337 

Cochran-Armitage test P=0.377 

Fisher exact test P=0.462 P=0.579N P=0.384 

Pituitary Gland (Pars Distalis): Adenoma 

Overall rate 24/50 (48%) 23/51 (45%) 18/50 (36%) 20/51 (39%) 

Adjusted rate 62.3% 64.9% 42.3% 55.0% 

Terminal rate 9/22 (41%) 14/26 (54%) 13/37 (35%) 8/23 (35%) 

First incidence (days) 537 601 610 448 

Life table test P=0.287N P=0.374N P=0.015N P=0.275N 

Logistic regression test P=0.255N P=0.484N P=0.146N P=0.269N 

Cochran-Armitage test P=0.232N 

Fisher exact test P=0.463N P=0.156N P=0.245N 

Pituitary Gland (Pars Distalis): Adenoma or Carcinoma 

Overall rate 24/50 (48%) 23/51 (45%) 18/50 (36%) 22/51 (43%) 

Adjusted rate 62.3% 64.9% 42.3% 59.4% 

Terminal rate 9/22 (41%) 14/26 (54%) 13/37 (35%) 9/23 (39%) 

First incidence (days) 537 601 610 448 

Life table test P=0.441N P=0.374N P=0.015N P=0.389N 

Logistic regression test P=0.429N P=0.484N P=0.146N P=0.419N 

Cochran-Armitage test P=0.395N 

Fisher exact test P=0.463N P=0.156N P=0.386N 

Stomach (Forestomach): Squamous Cell Papilloma or Squamous Cell Carcinoma 

Overall rate 0/50 (0%) 3/51 (6%) 0/50 (0%) 2/51 (4%) 

Adjusted rate 0.0% 11.5% 0.0% 6.5% 

Terminal rate 0/22 (0%) 3/26 (12%) 0/37 (0%) 1/23 (4%) 

First incidence (days) — 740 (T) — 590 

Life table test P=0.403 P=0.150 — P=0.247 

Logistic regression test P=0.420 P=0.150 — P=0.249 

Cochran-Armitage test P=0.426 

Fisher exact test P=0.125 — P=0.252 
D&C Yellow No. 11, NTP TR 463 127 

TABLE B3
 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Thyroid Gland (C-cell): Adenoma 

Overall rate 2/50 (4%) 2/51 (4%) 4/50 (8%) 6/51 (12%) 

Adjusted rate 9.1% 7.7% 10.5% 20.2% 

Terminal rate 2/22 (9%) 2/26 (8%) 3/37 (8%) 3/23 (13%) 

First incidence (days) 740 (T) 740 (T) 722 656 

Life table test P=0.050 P=0.635N P=0.572 P=0.159 

Logistic regression test P=0.059 P=0.635N P=0.475 P=0.140 

Cochran-Armitage test P=0.062 

Fisher exact test P=0.684N P=0.339 P=0.141 

Thyroid Gland (C-cell): Adenoma or Carcinoma 

Overall rate 2/50 (4%) 3/51 (6%) 5/50 (10%) 6/51 (12%) 

Adjusted rate 9.1% 11.5% 12.5% 20.2% 

Terminal rate 2/22 (9%) 3/26 (12%) 3/37 (8%) 3/23 (13%) 

First incidence (days) 740 (T) 740 (T) 638 656 

Life table test P=0.091 P=0.577 P=0.418 P=0.159 

Logistic regression test P=0.100 P=0.577 P=0.264 P=0.140 

Cochran-Armitage test P=0.105 

Fisher exact test P=0.509 P=0.218 P=0.141 

Thyroid Gland (Follicular Cell): Adenoma or Carcinoma 

Overall rate 0/50 (0%) 1/51 (2%) 1/50 (2%) 3/51 (6%) 

Adjusted rate 0.0% 3.8% 2.7% 10.1% 

Terminal rate 0/22 (0%) 1/26 (4%) 1/37 (3%) 1/23 (4%) 

First incidence (days) — 740 (T) 740 (T) 720 

Life table test P=0.059 P=0.533 P=0.604 P=0.143 

Logistic regression test P=0.063 P=0.533 P=0.604 P=0.126 

Cochran-Armitage test P=0.066 

Fisher exact test P=0.505 P=0.500 P=0.125 

Uterus: Stromal Polyp 

Overall rate 12/50 (24%) 11/51 (22%) 7/50 (14%) 6/51 (12%) 

Adjusted rate 33.3% 31.3% 17.4% 18.9% 

Terminal rate 2/22 (9%) 4/26 (15%) 5/37 (14%) 1/23 (4%) 

First incidence (days) 570 548 638 648 

Life table test P=0.078N P=0.454N P=0.050N P=0.110N 

Logistic regression test P=0.067N P=0.477N P=0.170N P=0.093N 

Cochran-Armitage test P=0.065N 

Fisher exact test P=0.478N P=0.154N P=0.089N 

All Organs: Mononuclear Cell Leukemia 

Overall rate 16/50 (32%) 21/51 (41%) 19/50 (38%) 16/51 (31%) 

Adjusted rate 47.3% 49.8% 41.9% 40.8% 

Terminal rate 6/22 (27%) 7/26 (27%) 11/37 (30%) 4/23 (17%) 

First incidence (days) 589 308 532 441 

Life table test P=0.365N P=0.308 P=0.345N P=0.533N 

Logistic regression test P=0.311N P=0.234 P=0.353 P=0.564N 

Cochran-Armitage test P=0.331N 

Fisher exact test P=0.227 P=0.338 P=0.558N 
128 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE B3
 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

All Organs: Benign Neoplasms 

Overall rate 42/50 (84%) 44/51 (86%) 41/50 (82%) 41/51 (80%) 

Adjusted rate 93.2% 93.5% 87.2% 91.1% 

Terminal rate 19/22 (86%) 23/26 (88%) 31/37 (84%) 19/23 (83%) 

First incidence (days) 537 548 456 448 

Life table test P=0.447N P=0.462N P=0.006N P=0.428N 

Logistic regression test P=0.424N P=0.420 P=0.372N P=0.565N 

Cochran-Armitage test P=0.294N 

Fisher exact test P=0.483 P=0.500N P=0.416N 

All Organs: Malignant Neoplasms 

Overall rate 30/50 (60%) 29/51 (57%) 30/50 (60%) 28/51 (55%) 

Adjusted rate 71.5% 64.1% 63.6% 64.4% 

Terminal rate 11/22 (50%) 11/26 (42%) 20/37 (54%) 9/23 (39%) 

First incidence (days) 481 308 456 441 

Life table test P=0.439N P=0.399N P=0.076N P=0.398N 

Logistic regression test P=0.429N P=0.432N P=0.510 P=0.364N 

Cochran-Armitage test P=0.370N 

Fisher exact test P=0.453N P=0.581N P=0.376N 

All Organs: Benign or Malignant Neoplasms 

Overall rate 48/50 (96%) 50/51 (98%) 50/50 (100%) 49/51 (96%) 

Adjusted rate 96.0% 98.0% 100.0% 98.0% 

Terminal rate 20/22 (91%) 25/26 (96%) 37/37 (100%) 22/23 (96%) 

First incidence (days) 481 308 456 441 

Life table test P=0.477 P=0.452N P=0.011N P=0.536N 

Logistic regression test P=0.607N P=0.441 P=0.133 P=0.593 

Cochran-Armitage test P=0.518N 

Fisher exact test P=0.492 P=0.247 P=0.684 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, clitoral gland, 

liver, lung, pituitary gland, thyroid gland, and uterus; for other tissues, denominator is number of animals necropsied. b Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality
 

Observed incidence at terminal kill
 d Beneath the control incidence are the P values associated with the trend test. Beneath the exposed group incidence are the P values corresponding to pairwise 

comparisons between the controls and that exposed group. The life table test regards neoplasms in animals dying prior to terminal kill as being (directly or indirectly) 

the cause of death. The logistic regression test regards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence 

rates. For all tests, a negative trend or a lower incidence in an exposure group is indicated byN. e Not applicable; no neoplasms in animal group 
D&C Yellow No. 11, NTP TR 463 129 

TABLE B4a 

Historical Incidence of Hepatocellular Neoplasms in Untreated Female F344/N Rats a 

Incidence in Controls 

Study Adenoma Carcinoma Adenoma or Carcinoma 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 0/50 0/50 0/50 

Benzyl Acetate 1/50 0/50 1/50 

Butyl Benzyl Phthalate 0/50 0/50 0/50 

C.I. Pigment Red 23 1/50 0/50 1/50 

C.I. Pigment Red 3 0/50 0/50 0/50 

o-Nitroanisole 0/50 0/50 0/50 

p-Nitrobenzoic Acid 2/50 0/50 2/50 

Polysorbate 80 0/50 0/50 0/50 

Overall Historical Incidence 

Total 8/1,301 (0.6%) 1/1,301 (0.1%) 9/1,301 (0.7%) 

Standard deviation 1.5% 0.4% 1.5% 

Range 0%-6% 0%-2% 0%-6% 

a Data as of 12 May 1995 

TABLE B4b 

Historical Incidence of Renal Tubule Neoplasms in Untreated Female F344/N Rats a 

Incidence in Controls 

Study Adenoma Carcinoma Adenoma or Carcinoma 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 0/50 0/50 0/50 

Benzyl Acetate 0/50 0/50 0/50 

Butyl Benzyl Phthalate 0/50 1/50 1/50 

C.I. Pigment Red 23 0/50 0/50 0/50 

C.I. Pigment Red 3 0/50 0/50 0/50 

o-Nitroanisole 0/50 0/50 0/50 

p-Nitrobenzoic Acid 0/50 0/50 0/50 

Polysorbate 80 0/50 0/50 0/50 

Overall Historical Incidence 

Total 0/1,298 (0%) 1/1,298 (0.1%) 1/1,298 (0.1%) 

Standard deviation 0.4% 0.4% 

Range 0%-2% 0%-2% 

a Data as of 12 May 1995 
130 D&C Yellow No. 11, NTP TR 463 

TABLE B4c 

Historical Incidence of Oral Cavity Neoplasms in Untreated Female F344/N Rats a 

Incidence in Controls 

Study Squamous Cell 

Papillomab 

Squamous Cell 

Carcinoma 

Squamous Cell 

Papilloma or Carcinomab 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 2/50 0/50 2/50 

Benzyl Acetate 1/50 0/50 1/50 

Butyl Benzyl Phthalate 2/50 0/50 2/50 

C.I. Pigment Red 23 0/50 0/50 0/50 

C.I. Pigment Red 3 0/50 0/50 0/50 

o-Nitroanisole 1/50 0/50 1/50 

p-Nitrobenzoic Acid 0/50 0/50 0/50 

Polysorbate 80 0/50 0/50 0/50 

Overall Historical Incidence 

Total 11/1,301 (0.9%) 4/1,301 (0.3%) 15/1,301 (1.2%) 

Standard Deviation 1.4% 0.7% 1.6% 

Range 0%-4% 0%-2% 0%-6% 

a Data as of 12 May 1995. Includes data for oral mucosa, tongue, pharynx, and tooth. b Includes data for papilloma. 

TABLE B4d 

Historical Incidence of Clitoral Gland Neoplasms in Untreated Female F344/N Rats a 

Incidence in Controls 

Study Adenoma Carcinoma Adenoma or Carcinoma 

Historical Incidence at Southern Research Institute 

2,2-Bis(bromomethyl)-1,3-propanediol (FR-1138®) 4/48 1/48 5/48 

Benzyl Acetate 0/50 1/50 1/50 

Butyl Benzyl Phthalate 3/50 4/50 7/50 

C.I. Pigment Red 23 5/47 3/47 7/47 

C.I. Pigment Red 3 9/47 0/47 9/47 

o-Nitroanisole 3/45 4/45 7/45 

p-Nitrobenzoic Acid 4/50 1/50 4/50 

Polysorbate 80 3/48 7/48 10/48 

Overall Historical Incidence 

Total 99/1,218 (8.1%) 33/1,218 (2.7%) 130/1,218 (10.7%) 

Standard deviation 4.1% 3.8% 5.3% 

Range 0%-19% 0%-15% 2%-21% 

a Data as of 12 May 1995 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 131 

TABLE B5 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 a 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Disposition Summary 

Animals initially in study 60 60 60 60 

12-Month interim evaluation 10 9 10 9 

Early deaths 

Moribund 25 23 12 25 

Natural deaths 3 2 1 3 

Survivors 

Terminal sacrifice 22 26 37 23 

Animals examined microscopically 60 60 60 60 

12-Month Interim Evaluation 

Alimentary System 

Intestine large, colon (10) (8) (9) (9) 

Parasite metazoan 1 (10%) 1 (11%) 

Intestine large, rectum (10) (9) (10) (9) 

Parasite metazoan 2 (20%) 1 (11%) 

Liver (10) (9) (10) (9) 

Basophilic focus 7 (70%) 2 (22%) 2 (20%) 1 (11%) 

Clear cell focus 1 (11%) 4 (40%) 4 (44%) 

Eosinophilic focus 1 (10%) 

Granuloma 1 (10%) 

Hepatodiaphragmatic nodule 1 (11%) 2 (20%) 

Inflammation, subacute 3 (30%) 4 (44%) 7 (70%) 9 (100%) 

Bile duct, hyperplasia 1 (10%) 1 (11%) 6 (60%) 9 (100%) 

Bile duct, pigmentation 9 (100%) 7 (70%) 9 (100%) 

Hepatocyte, cytologic alterations 4 (44%) 10 (100%) 9 (100%) 

Hepatocyte, pigmentation 9 (100%) 10 (100%) 9 (100%) 

Kupffer cell, pigmentation 9 (100%) 

Mesentery (1) (2) (2) 

Accessory spleen 2 (100%) 

Fat, necrosis 1 (100%) 2 (100%) 

Pancreas (10) (9) (10) (9) 

Atrophy 2 (20%) 3 (30%) 1 (11%) 

Salivary glands (10) (9) (10) (9) 

Atrophy 1 (10%) 2 (22%) 

Stomach, forestomach (10) (9) (10) (9) 

Mucosa, hyperplasia 1 (10%) 

Stomach, glandular (10) (9) (10) (9) 

Mineralization 2 (22%) 

Cardiovascular System 

Heart (10) (9) (10) (9) 

Cardiomyopathy 1 (10%) 

Endocrine System 

Adrenal cortex (10) (9) (10) (9) 

Accessory adrenal cortical nodule 4 (44%) 2 (20%) 1 (11%) 

Hypertrophy, focal 1 (10%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

132 D&C Yellow No. 11, NTP TR 463 

TABLE B5
 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

12-Month Interim Evaluation (continued) 

Endocrine System (continued) 

Pituitary gland (10) (9) (10) (9) 

Pars distalis, angiectasis 1 (10%) 1 (10%) 1 (11%) 

Pars distalis, cyst 4 (40%) 7 (78%) 9 (90%) 3 (33%) 

Pars distalis, hyperplasia, focal 1 (10%) 2 (22%) 2 (20%) 1 (11%) 

Pars intermedia, cyst 1 (10%) 

Pars intermedia, hyperplasia 1 (10%) 

Thyroid gland (10) (9) (10) (9) 

Ultimobranchial cyst 2 (22%) 1 (10%) 1 (11%) 

Genital System 

Clitoral gland (10) (9) (10) (9) 

Hyperplasia 1 (10%) 

Inflammation, chronic 1 (11%) 

Inflammation, chronic active 1 (11%) 

Ovary (10) (9) (10) (9) 

Cyst 1 (10%) 3 (33%) 6 (60%) 1 (11%) 

Uterus (10) (9) (10) (9) 

Hydrometra 1 (11%) 1 (10%) 3 (33%) 

Hematopoietic System 

Lymph node (1) (2) (2) (2) 

Mediastinal, hemorrhage 1 (100%) 2 (100%) 2 (100%) 2 (100%) 

Mediastinal, pigmentation 1 (100%) 2 (100%) 2 (100%) 2 (100%) 

Lymph node, mandibular (10) (9) (10) (9) 

Hemorrhage 3 (30%) 1 (10%) 

Pigmentation 1 (10%) 4 (44%) 1 (10%) 2 (22%) 

Spleen (10) (9) (10) (9) 

Hematopoietic cell proliferation 1 (10%) 1 (11%) 

Pigmentation 7 (70%) 7 (78%) 6 (60%) 6 (67%) 

Integumentary System 

Mammary gland (10) (9) (10) (9) 

Hyperplasia 1 (10%) 1 (10%) 

Respiratory System 

Lung (10) (9) (10) (9) 

Hemorrhage 1 (10%) 

Infiltration cellular, histiocyte 4 (40%) 1 (11%) 1 (10%) 2 (22%) 

Inflammation, subacute 3 (33%) 1 (10%) 2 (22%) 

Alveolar epithelium, hyperplasia 1 (10%) 

Special Senses System 

Eye (1) 

Cataract 1 (100%) 

Hemorrhage 1 (100%) 

Retina, degeneration 1 (100%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 133 

TABLE B5
 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

12-Month Interim Evaluation (continued) 

Urinary System 

Kidney (10) (9) (10) (9) 

Mineralization 10 (100%) 9 (100%) 7 (70%) 9 (100%) 

Nephropathy 6 (60%) 6 (67%) 7 (70%) 9 (100%) 

Renal tubule, pigmentation 9 (100%) 10 (100%) 9 (100%) 

Transitional epithelium, hyperplasia 2 (22%) 

Systems Examined With No Lesions Observed 

General Body System 

Musculoskeletal System 

Nervous System 

2-Year Study 

Alimentary System 

Intestine large, colon (50) (50) (50) (51) 

Parasite metazoan 4 (8%) 4 (8%) 2 (4%) 

Intestine large, rectum (49) (50) (49) (50) 

Edema 1 (2%) 

Parasite metazoan 3 (6%) 4 (8%) 6 (12%) 2 (4%) 

Intestine large, cecum (50) (51) (49) (51) 

Edema 1 (2%) 1 (2%) 2 (4%) 

Parasite metazoan 1 (2%) 

Ulcer 1 (2%) 

Intestine small, duodenum (50) (51) (50) (50) 

Ulcer 1 (2%) 

Epithelium, hyperplasia 1 (2%) 

Intestine small, jejunum (49) (51) (50) (51) 

Epithelium, hyperplasia 1 (2%) 

Intestine small, ileum (48) (51) (49) (49) 

Epithelium, hyperplasia 1 (2%) 

Liver (50) (51) (50) (51) 

Angiectasis 1 (2%) 2 (4%) 3 (6%) 1 (2%) 

Basophilic focus 32 (64%) 26 (51%) 11 (22%) 12 (24%) 

Clear cell focus 10 (20%) 18 (35%) 29 (58%) 30 (59%) 

Cyst 3 (6%) 4 (8%) 

Cytoplasmic alteration 2 (4%) 

Degeneration, cystic 1 (2%) 

Eosinophilic focus 10 (20%) 9 (18%) 14 (28%) 16 (31%) 

Granuloma 10 (20%) 1 (2%) 1 (2%) 1 (2%) 

Hematopoietic cell proliferation 2 (4%) 1 (2%) 2 (4%) 7 (14%) 

Hepatodiaphragmatic nodule 7 (14%) 4 (8%) 5 (10%) 5 (10%) 

Inflammation, subacute 2 (4%) 1 (2%) 3 (6%) 3 (6%) 

Mixed cell focus 12 (24%) 19 (37%) 20 (40%) 16 (31%) 

Necrosis, focal 2 (4%) 2 (4%) 5 (10%) 4 (8%) 

Thrombosis 1 (2%) 2 (4%) 2 (4%) 

Bile duct, hyperplasia 14 (28%) 10 (20%) 27 (54%) 33 (65%) 

Bile duct, pigmentation 46 (90%) 49 (98%) 50 (98%) 

Centrilobular, atrophy 4 (8%) 9 (18%) 5 (10%) 5 (10%) 

Centrilobular, necrosis 1 (2%) 1 (2%) 

Hepatocyte, cytologic alterations 11 (22%) 31 (62%) 40 (78%) 

Hepatocyte, pigmentation 34 (67%) 44 (88%) 50 (98%) 

Hepatocyte, vacuolization cytoplasmic 6 (12%) 6 (12%) 2 (4%) 4 (8%) 

Kupffer cell, pigmentation 9 (18%) 11 (22%) 16 (32%) 32 (63%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

134 D&C Yellow No. 11, NTP TR 463 

TABLE B5
 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Alimentary System (continued) 

Mesentery (11) (18) (8) (12) 

Accessory spleen 1 (9%) 

Angiectasis 1 (9%) 

Fat, necrosis 8 (73%) 16 (89%) 8 (100%) 11 (92%) 

Pancreas (50) (51) (50) (51) 

Atrophy 9 (18%) 19 (37%) 17 (34%) 14 (27%) 

Necrosis 1 (2%) 

Acinar cell, cytoplasmic alteration 3 (6%) 2 (4%) 2 (4%) 

Acinar cell, hyperplasia, focal 1 (2%) 1 (2%) 1 (2%) 

Salivary glands (50) (46) (50) (50) 

Atrophy 8 (17%) 7 (14%) 13 (26%) 

Stomach, forestomach (50) (51) (50) (51) 

Edema 5 (10%) 2 (4%) 4 (8%) 

Fibrosis 1 (2%) 

Ulcer 4 (8%) 1 (2%) 3 (6%) 

Mucosa, hyperplasia 5 (10%) 17 (33%) 30 (60%) 27 (53%) 

Stomach, glandular (50) (51) (50) (51) 

Edema 2 (4%) 2 (4%) 2 (4%) 

Erosion 2 (4%) 1 (2%) 2 (4%) 

Ulcer 1 (2%) 2 (4%) 

Tooth (1) 

Developmental malformation 1 (100%) 

Cardiovascular System 

Blood vessel (50) (51) (50) (51) 

Hypertrophy 1 (2%) 

Heart (50) (51) (50) (51) 

Cardiomyopathy 16 (32%) 18 (35%) 18 (36%) 12 (24%) 

Mineralization 1 (2%) 

Thrombosis 1 (2%) 

Endocrine System 

Adrenal cortex (50) (51) (50) (51) 

Accessory adrenal cortical nodule 5 (10%) 13 (25%) 11 (22%) 13 (25%) 

Angiectasis 2 (4%) 2 (4%) 

Degeneration, fatty 9 (18%) 10 (20%) 9 (18%) 5 (10%) 

Hematopoietic cell proliferation 1 (2%) 1 (2%) 1 (2%) 

Hyperplasia, diffuse 1 (2%) 1 (2%) 

Hyperplasia, focal 3 (6%) 2 (4%) 2 (4%) 1 (2%) 

Hypertrophy 2 (4%) 

Hypertrophy, focal 3 (6%) 2 (4%) 2 (4%) 

Necrosis 1 (2%) 1 (2%) 

Adrenal medulla (48) (51) (50) (51) 

Hyperplasia 3 (6%) 1 (2%) 4 (8%) 

Islets, pancreatic (50) (51) (49) (51) 

Hyperplasia 1 (2%) 1 (2%) 

Parathyroid gland (48) (48) (48) (50) 

Hyperplasia 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 135 

TABLE B5
 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Endocrine System (continued) 

Pituitary gland (50) (51) (50) (51) 

Pars distalis, angiectasis 10 (20%) 6 (12%) 16 (32%) 12 (24%) 

Pars distalis, cyst 25 (50%) 21 (41%) 25 (50%) 23 (45%) 

Pars distalis, hyperplasia, focal 9 (18%) 4 (8%) 8 (16%) 8 (16%) 

Pars intermedia, angiectasis 3 (6%) 1 (2%) 1 (2%) 

Pars intermedia, cyst 2 (4%) 2 (4%) 

Pars intermedia, hyperplasia 1 (2%) 

Thyroid gland (50) (51) (50) (51) 

Ultimobranchial cyst 3 (6%) 5 (10%) 4 (8%) 4 (8%) 

C-cell, hyperplasia 13 (26%) 7 (14%) 10 (20%) 9 (18%) 

Follicle, cyst 4 (8%) 4 (8%) 

Follicular cell, hyperplasia 1 (2%) 

General Body System 

None 

Genital System 

Clitoral gland (49) (50) (49) (51) 

Cyst 2 (4%) 4 (8%) 3 (6%) 1 (2%) 

Hyperplasia 3 (6%) 1 (2%) 1 (2%) 1 (2%) 

Inflammation, chronic 1 (2%) 

Inflammation, suppurative 2 (4%) 1 (2%) 

Ovary (50) (51) (50) (51) 

Cyst 13 (26%) 14 (27%) 11 (22%) 18 (35%) 

Uterus (50) (51) (50) (51) 

Hydrometra 2 (4%) 8 (16%) 3 (6%) 7 (14%) 

Hyperplasia, cystic 5 (10%) 1 (2%) 4 (8%) 

Hematopoietic System 

Bone marrow (50) (51) (50) (51) 

Hyperplasia 4 (8%) 1 (2%) 3 (6%) 5 (10%) 

Infiltration cellular, histiocyte 1 (2%) 

Myelofibrosis 3 (6%) 2 (4%) 3 (6%) 3 (6%) 

Necrosis 1 (2%) 

Lymph node (9) (11) (11) (15) 

Deep cervical, pigmentation 1 (9%) 

Iliac, pigmentation 1 (7%) 

Inguinal, hyperplasia, lymphoid 1 (11%) 

Mediastinal, hemorrhage 1 (11%) 2 (18%) 3 (27%) 2 (13%) 

Mediastinal, hyperplasia, lymphoid 2 (18%) 1 (9%) 1 (7%) 

Mediastinal, pigmentation 5 (56%) 7 (64%) 6 (55%) 8 (53%) 

Pancreatic, granuloma 1 (7%) 

Pancreatic, hemorrhage 1 (11%) 2 (13%) 

Pancreatic, hyperplasia, lymphoid 1 (11%) 1 (9%) 2 (13%) 

Pancreatic, pigmentation 1 (11%) 2 (18%) 6 (55%) 8 (53%) 

Renal, hemorrhage 1 (9%) 1 (7%) 

Renal, hyperplasia, lymphoid 1 (7%) 

Renal, pigmentation 1 (9%) 1 (7%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

136 D&C Yellow No. 11, NTP TR 463 

TABLE B5
 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Hematopoietic System (continued) 

Lymph node, mandibular (50) (51) (50) (49) 

Ectasia 1 (2%) 3 (6%) 7 (14%) 5 (10%) 

Hemorrhage 10 (20%) 11 (22%) 16 (32%) 7 (14%) 

Hyperplasia, lymphoid 5 (10%) 10 (20%) 14 (28%) 13 (27%) 

Pigmentation 23 (46%) 24 (47%) 20 (40%) 19 (39%) 

Lymph node, mesenteric (50) (51) (50) (51) 

Hemorrhage 5 (10%) 1 (2%) 1 (2%) 2 (4%) 

Hyperplasia, lymphoid 2 (4%) 5 (10%) 5 (10%) 2 (4%) 

Pigmentation 1 (2%) 1 (2%) 

Spleen (50) (50) (50) (51) 

Congestion 1 (2%) 

Developmental malformation 1 (2%) 

Fibrosis 3 (6%) 5 (10%) 4 (8%) 

Hematopoietic cell proliferation 27 (54%) 23 (46%) 30 (60%) 31 (61%) 

Infiltration cellular, histiocyte 1 (2%) 

Metaplasia, osseous 1 (2%) 

Necrosis 1 (2%) 1 (2%) 1 (2%) 

Pigmentation 30 (60%) 30 (60%) 26 (52%) 32 (63%) 

Lymphoid follicle, atrophy 2 (4%) 

Thymus (50) (49) (49) (49) 

Hemorrhage 1 (2%) 

Hyperplasia 1 (2%) 

Integumentary System 

Mammary gland (50) (51) (50) (51) 

Hyperplasia 46 (92%) 42 (82%) 36 (72%) 42 (82%) 

Skin (49) (51) (50) (51) 

Subcutaneous tissue, edema 1 (2%) 

Musculoskeletal System 

Bone (50) (51) (50) (51) 

Arthrosis 1 (2%) 

Fibrous osteodystrophy 1 (2%) 

Hyperostosis 1 (2%) 

Cranium, osteopetrosis 6 (12%) 7 (14%) 3 (6%) 3 (6%) 

Femur, osteopetrosis 5 (10%) 7 (14%) 1 (2%) 2 (4%) 

Nervous System 

Brain (50) (51) (50) (51) 

Developmental malformation 9 (18%) 11 (22%) 8 (16%) 14 (27%) 

Gliosis 1 (2%) 

Hemorrhage 1 (2%) 

Hydrocephalus 2 (4%) 4 (8%) 

Peripheral nerve (2) (2) (2) 

Degeneration 1 (50%) 

Inflammation, chronic 1 (50%) 

Spinal cord (3) (2) (2) 

Necrosis 1 (33%) 
 

 

 

 

 

 

 

 

 

D&C Yellow No. 11, NTP TR 463 137 

TABLE B5
 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 (continued)
 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

2-Year Study (continued) 

Respiratory System 

Lung (50) (51) (50) (51) 

Edema 1 (2%) 

Hemorrhage 1 (2%) 6 (12%) 

Infiltration cellular, histiocyte 34 (68%) 33 (65%) 36 (72%) 41 (80%) 

Inflammation, subacute 3 (6%) 2 (4%) 

Alveolar epithelium, hyperplasia 2 (4%) 3 (6%) 6 (12%) 

Nose (50) (51) (50) (51) 

Exudate 5 (10%) 5 (10%) 3 (6%) 2 (4%) 

Foreign body 2 (4%) 1 (2%) 2 (4%) 

Mucosa, hyperplasia 4 (8%) 2 (4%) 2 (4%) 

Mucosa, metaplasia, squamous 3 (6%) 2 (4%) 2 (4%) 1 (2%) 

Special Senses System 

Eye (1) (2) 

Atrophy 1 (100%) 

Cataract 2 (100%) 

Retina, degeneration 2 (100%) 

Urinary System 

Kidney (50) (51) (50) (51) 

Inflammation, suppurative 1 (2%) 

Mineralization 48 (96%) 49 (96%) 47 (94%) 33 (65%) 

Nephropathy 45 (90%) 47 (92%) 46 (92%) 50 (98%) 

Renal tubule, cytoplasmic alteration 1 (2%) 2 (4%) 1 (2%) 1 (2%) 

Renal tubule, necrosis 1 (2%) 1 (2%) 

Renal tubule, pigmentation 10 (20%) 48 (94%) 50 (100%) 51 (100%) 

Transitional epithelium, hyperplasia 2 (4%) 6 (12%) 10 (20%) 3 (6%) 

Urinary bladder (50) (51) (50) (51) 

Hyperplasia 1 (2%) 
138 D&C Yellow No. 11, NTP TR 463 
139 

APPENDIX C
 

GENETIC TOXICOLOGY
 

SALMONELLA MUTAGENICITY TEST PROTOCOL ..................................................... 140
 

CHINESE HAMSTER OVARY CELL CYTOGENETICS PROTOCOLS ........................................ 140
 

MOUSE PERIPHERAL BLOOD MICRONUCLEUS TEST PROTOCOL ....................................... 141
 

RESULTS .................................................................................... 142
 

TABLE C1 Mutagenicity of D&C Yellow No. 11 in Salmonella typhimurium ........................ 143
 

TABLE C2 Induction of Sister Chromatid Exchanges in Chinese Hamster Ovary Cells 

by D&C Yellow No. 11 ............................................................ 150
 

TABLE C3 Induction of Chromosomal Aberrations in Chinese Hamster Ovary Cells 

by D&C Yellow No. 11 ............................................................ 151
 

TABLE C4 Frequency of Micronuclei in Mouse Peripheral Blood Erythrocytes
 

Following Treatment with D&C Yellow No. 11 in Feed for 13 Weeks ..................... 152
 
 

140 D&C Yellow No. 11, NTP TR 463 

GENETIC TOXICOLOGY 

SALMONELLA MUTAGENICITY TEST PROTOCOL 

Testing was performed as reported by Zeiger et al. (1988). D&C Yellow No. 11 was sent to the laboratories 

as a coded aliquot from Radian Corporation (Austin, TX). It was incubated with the Salmonella 

typhimurium tester strains TA97, TA98, TA100, TA1535, and TA1538 either in buffer or S9 mix 

(metabolic activation enzymes and cofactors from Aroclor 1254-induced male Sprague-Dawley rat or Syrian 

hamster liver) for 20 minutes at 37 C. Top agar supplemented with l-histidine and d-biotin was added, and 

the contents of the tubes were mixed and poured onto the surfaces of minimal glucose agar plates. Histidineindependent mutant colonies arising on these plates were counted following incubation for 2 days at 37 C. 

Each trial consisted of triplicate plates of concurrent positive and negative controls and at least five doses of 

D&C Yellow No. 11. By study design, 10,000 μg/plate was selected as the high dose in the study conducted 

at SRI International, and 4,000 μg/plate was selected in the study conducted at Microbiological Associates, 

Inc. All positive trials were repeated under the conditions that elicited the positive response. 

In this assay, a positive response is defined as a reproducible, dose-related increase in histidine-independent 

(revertant) colonies in any one strain/activation combination. An equivocal response is defined as an increase 

in revertants that is not dose related, is not reproducible, or is of insufficient magnitude to support a 

determination of mutagenicity. A negative response is obtained when no increase in revertant colonies is 

observed following chemical treatment. There is no minimum percentage or fold increase required for a 

chemical to be judged positive or weakly positive. 

CHINESE HAMSTER OVARY CELL CYTOGENETICS PROTOCOLS 

Testing was performed as reported by Galloway et al. (1987). D&C Yellow No. 11 was sent to the 

laboratory as a coded aliquot by Radian Corporation. It was tested in cultured Chinese hamster ovary (CHO) 

cells for induction of sister chromatid exchanges (SCEs) and chromosomal aberrations (Abs), both in the 

presence and absence of Aroclor 1254-induced male Sprague-Dawley rat liver S9 and cofactor mix. Cultures 

were handled under gold lights to prevent photolysis of bromodeoxyuridine-substituted DNA. Each test 

consisted of concurrent solvent and positive controls and of at least three doses of D&C Yellow No. 11; the 

high dose was limited by toxicity. A single flask per dose was used. 

Sister Chromatid Exchange Test: In the SCE test without S9, CHO cells were incubated for 26 hours with 

D&C Yellow No. 11 in McCoy’s 5A medium supplemented with fetal bovine serum, l-glutamine, and 

antibiotics. Bromodeoxyuridine (BrdU) was added 2 hours after culture initiation. After 26 hours, the 

medium containing D&C Yellow No. 11 was removed and replaced with fresh medium plus BrdU and 

Colcemid, and incubation was continued for 2 hours. Cells were then harvested by mitotic shake-off, fixed, 

and stained with Hoechst 33258 and Giemsa. In the SCE test with S9, cells were incubated with D&C 

Yellow No. 11, serum-free medium, and S9 for 2 hours. The medium was then removed and replaced with 

medium containing serum and BrdU and no D&C Yellow No. 11, and incubation proceeded for an additional 

26 hours with Colcemid present for the final 2 hours. Harvesting and staining were the same as for cells 

treated without S9. All slides were scored blind, and those from a single test were read by the same person. 

Fifty second-division metaphase cells were scored for frequency of SCEs/cell from each dose level. Because 

significant chemical-induced cell cycle delay was seen at some dose levels, incubation time was lengthened 

for these cultures to ensure a sufficient number of scorable (second-division metaphase) cells. 
 

D&C Yellow No. 11, NTP TR 463 141 

Statistical analyses were conducted on the slopes of the dose-response curves and the individual dose points 

(Galloway et al., 1987). An SCE frequency 20% above the concurrent solvent control value was chosen as a 

statistically conservative positive response. The probability of this level of difference occurring by chance at 

one dose point is less than 0.01; the probability for such a chance occurrence at two dose points is less than 

0.001. An increase of 20% or greater at any single dose was considered weak evidence of activity; increases 

at two or more doses resulted in a determination that the trial was positive. A statistically significant trend 

(P<0.005) in the absence of any responses reaching 20% above background led to a call of equivocal. 

Chromosomal Aberrations Test: In the Abs test without S9, cells were incubated in McCoy’s 5A medium 

with D&C Yellow No. 11 for 8.5 hours; Colcemid was added, and incubation continued for 2 hours. The 

cells were then harvested by mitotic shake-off, fixed, and stained with Giemsa. For the Abs test with S9, 

cells were treated with D&C Yellow No. 11 and S9 for 2 hours, after which the treatment medium was 

removed and the cells were incubated for 19.5 hours in fresh medium with Colcemid present for the final 

2 hours. Cells were harvested in the same manner as for the treatment without S9. The harvest time for the 

Abs test was based on the cell cycle information obtained in the SCE test; because cell cycle delay was 

anticipated, the incubation period was extended. 

Cells were selected for scoring on the basis of good morphology and completeness of karyotype 

(21 ± 2 chromosomes). All slides were scored blind, and those from a single test were read by the same 

person. Two hundred first-division metaphase cells were scored in the low-dose control cultures and the 

lowest dose in the initial trial without S9. Because high numbers of aberrations were observed, making a 

smaller sample size necessary for statistical precision and making the scoring process difficult, fewer cells 

(25 to 100) were scored in the other cultures. Classes of aberrations included simple (breaks and terminal 

deletions), complex (rearrangements and translocations), and other (pulverized cells, despiralized 

chromosomes, and cells containing 10 or more aberrations). 

Chromosomal aberration data are presented as percentages of cells with aberrations. To arrive at a statistical 

call for a trial, analyses were conducted on both the dose-response curve and individual dose points. For a 

single trial, a statistically significant (P 0.05) difference for one dose point and a significant trend (P 0.015) 

were considered weak evidence for a positive response; significant differences for two or more doses 

indicated the trial was positive. A positive trend test in the absence of a statistically significant increase at 

any one dose resulted in an equivocal call (Galloway et al., 1987). Ultimately, the trial calls were based on a 

consideration of the statistical analyses as well as the biological information available to the reviewers. 

MOUSE PERIPHERAL BLOOD MICRONUCLEUS TEST PROTOCOL 

A detailed discussion of this assay is presented in MacGregor et al. (1990). At the end of a 13-week toxicity 

study (NTP, 1991a), peripheral blood samples were obtained from male and female B6C3F 1 mice, and 

smears were immediately prepared and fixed in absolute methanol. The methanol-fixed slides were stained 

with a chromatin-specific fluorescent dye mixture of Hoechst 33258/pyronin Y (MacGregor et al., 1983) and 

coded. Slides were scanned at 630 or 1,000× magnification using a semi-automated image analysis system to 

determine the frequency of micronuclei in 10,000 normochromatic erythrocytes (NCEs) in each of 9 or 10 

animals per exposure group. The criteria of Schmid (1976) were used to define micronuclei, with the 

additional requirement that the micronuclei exhibit the characteristic fluorescent emissions of DNA (blue with 

360 nm and orange with 540 nm ultraviolet illumination); the minimum size was approximately 

one-twentieth the diameter of the NCE. 
142 D&C Yellow No. 11, NTP TR 463 

Log transformation of the NCE data, testing of normality by the Shapiro-Wilk test, and testing for 

heterogeneity of variance by Cochran’s test were performed before statistical analyses. The frequency of 

micronucleated cells among NCEs was analyzed by analysis of variance using the SAS GLM procedure. The 

NCE data for each exposure group were compared with the concurrent control using Student’s t-test. 

RESULTS 

Results of mutagenicity tests with D&C Yellow No. 11 in S. typhimurium were equivocal in the SRI 

International study, based on the responses observed in strain TA100 with 10% induced rat liver S9, and the 

results were weakly positive in the Microbiological Associates, Inc., study, which used slightly lower doses, 

based on responses observed in strains TA98 and TA100 with 30% induced rat or hamster liver S9 

(Table C1; Zeiger et al., 1988). No indication of mutagenic activity was observed in the absence of S9 in any 

of the strains tested. The data from the S. typhimurium studies indicate variable responses among replicate 

trials within a particular treatment condition; this may have been the result of precipitate formation at higher 

concentrations (333 μg/plate and above) and consequent variability in the actual D&C Yellow No. 11 

exposure concentrations. 

In cytogenic tests with cultured CHO cells, D&C Yellow No. 11 induced highly significant increases in both 

SCEs (Table C2) and Abs (Table C3) with and without S9. Cell cycle delay, requiring an extended 

incubation period, was observed in the SCE test at doses of 1.5 μg/mL and above; in the Abs test, no delay 

was observed in the absence of S9, but cultures treated in the presence of S9 were harvested late because cell 

cycle delay was anticipated. Less than 200 cells per dose level were scored at all but one dose level in the 

Abs test due to the high number of Abs per cell (cultures treated with S9), the frequency of aberrant cells, and 

the difficulty in finding scorable cells in some cases (Trial 1, without S9). 

Despite the strong response seen in the in vitro Abs assay, no increase in the frequency of micronucleated 

NCEs was observed in peripheral blood samples from male and female mice given D&C Yellow No. 11 in 

feed for 13 weeks (Table C4). 

In conclusion, D&C Yellow No. 11 was mutagenic in bacteria and clastogenic in mammalian cells in vitro, 

but no evidence of clastogenicity was observed in the single in vivo study performed with male and female 

mice. 
D&C Yellow No. 11, NTP TR 463 143 

TABLE C1 

Mutagenicity of D&C Yellow No. 11 in Salmonella typhimuriuma 

Revertants/plateb 

Strain Dose S9 +hamster S9 

(μg/plate) Trial 1 Trial 2 Trial 3 10% 10% 30% 

Study performed at SRI International 

TA100 0 

1 

3 

6 

10 

33 

100 

333 

1,000 

3,333 

10,000 

120 ± 7.4 

118 ± 7.1 

95 ± 5.2c 

91 ± 4.8c 

93 ± 5.8c 

111 ± 3.2c 

109 ± 5.8 

92 ± 10.7 

96 ± 13.1 

87 ± 11.6 

104 ± 10.4 

121 ± 8.2 

109 ± 11.0 

94 ± 5.8 

110 ± 5.9 

138 ± 7.8 

95 ± 6.7 

80 ± 5.9c 

132 ± 14.1 

169 ± 2.9 

151 ± 10.9c 

136 ± 8.1c 

158 ± 7.5c 

169 ± 6.4c 

115 ± 5.8 

114 ± 13.3 

116 ± 4.2 

112 ± 10.2 

128 ± 6.7 

148 ± 5.0 

119 ± 7.4 

133 ± 16.6 

133 ± 9.9 

157 ± 6.9 

169 ± 10.3 

131 ± 6.8c 

Trial summary 

Positive controld Negative 629 ± 23.5 

Negative 

438 ± 19.3 

Negative 

554 ± 21.5 

Negative 

683 ± 31.0 

Negative 

989 ± 51.7 

Equivocal 

622 ± 79.8 

+rat S9 

5% 5% 10% 10% 10% 10% 

TA100 0 

(continued) 0.3 

1 

3 

10 

16 

20 

33 

66 

100 

166 

200 

333 

1,000 

3,333 

10,000 

121 ± 6.5 

167 ± 5.9 

141 ± 18.9c 

153 ± 6.1c 

150 ± 10.9c 

161 ± 6.2c 

91 ± 5.9 

108 ± 11.2 

87 ± 8.1 

101 ± 8.1 

121 ± 25.2 

142 ± 16.6 

95 ± 7.8 

196 ± 10.8 

164 ± 7.6c 

154 ± 10.6c 

186 ± 8.8c 

202 ± 8.7c 

147 ± 4.8 

127 ± 13.0 

146 ± 4.5 

208 ± 21.2 

164 ± 16.9 

142 ± 7.5c 

143 ± 10.4 

178 ± 14.3 

182 ± 9.8c 

185 ± 6.8c 

184 ± 7.5c 

173 ± 7.1c 

144 ± 7.0 

181 ± 14.5 

208 ± 11.0 

211 ± 16.7 

184 ± 7.2 

182 ± 17.5 

186 ± 8.5 

182 ± 9.0c 

173 ± 7.0c 

165 ± 9.7c 

Trial summary 

Positive control 

Negative 

1,276 ± 45.0 

Equivocal 

889 ± 53.5 

Equivocal 

445 ± 9.2 

Equivocal 

470 ± 14.8 

Equivocal 

502 ± 24.0 

Equivocal 

776 ± 21.9 
144 D&C Yellow No. 11, NTP TR 463 

TABLE C1
 

Mutagenicity of D&C Yellow No. 11 in Salmonella typhimurium (continued)
 

Revertants/plate 

Strain Dose +rat S9 (continued) 

(μg/plate) 10% 10% 10% 30% 30% 30% 

Study performed at SRI International (continued) 

TA100 0 

(continued) 0.3 

1 

3 

6 

10 

33 

100 

333 

1,000 

3,333 

10,000 

135 ± 2.7 

116 ± 1.3 

115 ± 7.7 

151 ± 5.5 

200 ± 16.3 

213 ± 11.1 

113 ± 10.7 

115 ± 11.2 

121 ± 4.8 

115 ± 5.9 

151 ± 11.3 

181 ± 5.0 

86 ± 5.0 

88 ± 6.8 

79 ± 3.8 

93 ± 4.3 

112 ± 9.0 

142 ± 13.3 

141 ± 5.5 

159 ± 5.0 

164 ± 2.5c 

174 ± 12.0c 

149 ± 8.0c 

169 ± 3.4c 

112 ± 6.1 

97 ± 9.1 

90 ± 7.6 

84 ± 1.5 

85 ± 4.3 

120 ± 19.7 

112 ± 12.2 

94 ± 4.4 

107 ± 5.2 

123 ± 6.2 

178 ± 10.6 

154 ± 14.0c 

Trial summary 

Positive control 

Equivocal 

114 ± 4.5 

Equivocal 

567 ± 5.6 

Equivocal 

421 ± 27.9 

Negative 

307 ± 20.8 

Negative 

296 ± 11.0 

Equivocal 

245 ± 12.2 
D&C Yellow No. 11, NTP TR 463 145 

TABLE C1
 

Mutagenicity of D&C Yellow No. 11 in Salmonella typhimurium (continued)
 

Revertants/plate 

Strain Dose S9 +hamster S9 

(μg/plate) Trial 1 Trial 2 Trial 3 10% 10% 30% 

Study performed at SRI International (continued) 

TA1535 0 

1 

3 

6 

10 

33 

100 

333 

1,000 

3,333 

10,000 

16 ± 1.5 

6 ± 0.9 

8 ± 1.5c 

7 ± 0.6c 

6 ± 0.3c 

8 ± 3.2c 

16 ± 1.5 

17 ± 3.3 

17 ± 3.8 

15 ± 0.0 

15 ± 1.5 

20 ± 2.3 

21 ± 0.3 

14 ± 2.7 

15 ± 3.2 

13 ± 2.3 

11 ± 1.0 

9 ± 1.8c 

7 ± 1.2 

8 ± 1.2 

6 ± 0.7c 

7 ± 2.0c 

9 ± 0.3c 

8 ± 2.0c 

8 ± 2.0 

9 ± 2.1 

13 ± 2.6 

9 ± 2.3 

13 ± 0.3 

10 ± 0.9 

10 ± 1.8 

7 ± 0.6 

6 ± 1.2 

8 ± 1.8 

7 ± 0.3 

5 ± 1.2c 

Trial summary 

Positive control 

Negative 

434 ± 11.3 

Negative 

417 ± 13.9 

Negative 

442 ± 7.5 

Negative 

333 ± 31.3 

Negative 

483 ± 10.7 

Negative 

432 ± 55.5 

+rat S9 

10% 10% 30% 

TA1535 0 

(continued) 1 

3 

6 

10 

33 

100 

333 

1,000 

3,333 

10,000 

9 ± 1.5 

11 ± 2.9 

5 ± 0.6c 

5 ± 1.2c 

9 ± 1.3c 

10 ± 1.5c 

10 ± 2.3 

11 ± 1.8 

11 ± 1.5 

11 ± 1.9 

9 ± 1.3 

8 ± 0.6 

13 ± 2.0 

10 ± 2.0 

8 ± 1.8 

10 ± 2.7 

12 ± 1.5 

7 ± 0.7c 

Trial summary 

Positive control 

Negative 

158 ± 7.4 

Negative 

290 ± 5.5 

Negative 

97 ± 10.5 
146 D&C Yellow No. 11, NTP TR 463 

TABLE C1
 

Mutagenicity of D&C Yellow No. 11 in Salmonella typhimurium (continued)
 

Revertants/plate 

Strain Dose S9 +hamster S9 

(μg/plate) Trial 1 Trial 2 Trial 3 10% 10% 30% 

Study performed at SRI International (continued) 

TA97 0 

1 

3 

6 

10 

33 

100 

333 

1,000 

3,333 

10,000 

148 ± 4.7 

154 ± 4.3 

129 ± 3.4c 

113 ± 2.6c 

111 ± 14.2c 

141 ± 13.6c 

129 ± 7.9 

115 ± 9.8 

118 ± 10.7 

112 ± 7.2 

109 ± 16.7 

112 ± 15.6 

137 ± 10.9 

150 ± 8.6 

131 ± 10.5 

147 ± 10.8 

98 ± 7.0 

66 ± 25.5c 

189 ± 7.0 

202 ± 9.1 

169 ± 8.7c 

168 ± 17.9c 

209 ± 3.8c 

197 ± 5.7c 

158 ± 6.1 

167 ± 4.7 

168 ± 5.2 

166 ± 3.4 

176 ± 6.1 

174 ± 9.7 

159 ± 12.5 

153 ± 3.2 

182 ± 21.5 

211 ± 4.0 

181 ± 35.9 

109 ± 17.8c 

Trial summary 

Positive control 

Negative 

352 ± 22.5 

Negative 

579 ± 5.7 

Negative 

795 ± 35.9 

Negative 

1,060 ± 75.7 

Negative 

528 ± 26.9 

Negative 

536 ± 4.9 

+rat S9 

10% 10% 30% 

TA97 0 

(continued) 1 

3 

6 

10 

33 

100 

333 

1,000 

3,333 

10,000 

200 ± 5.8 

192 ± 3.4 

171 ± 29.9c 

147 ± 14.7c 

186 ± 9.8c 

187 ± 25.0c 

160 ± 2.9 

159 ± 10.7 

173 ± 5.5 

169 ± 2.2 

185 ± 12.8 

217 ± 22.6 

210 ± 8.1 

184 ± 14.1 

175 ± 7.4 

209 ± 5.8 

224 ± 5.1 

114 ± 4.0c 

Trial summary 

Positive control 

Negative 

365 ± 17.8 

Equivocal 

375 ± 13.5 

Negative 

469 ± 11.3 
D&C Yellow No. 11, NTP TR 463 147 

TABLE C1
 

Mutagenicity of D&C Yellow No. 11 in Salmonella typhimurium (continued)
 

Revertants/plate 

Strain Dose S9 +hamster S9 

(μg/plate) Trial 1 Trial 2 Trial 3 10% 10% 30% 

Study performed at SRI International (continued) 

TA98 0 

1 

3 

6 

10 

33 

100 

333 

1,000 

3,333 

10,000 

17 ± 3.5 

17 ± 3.3 

10 ± 1.2c 

11 ± 2.9c 

12 ± 0.7c 

19 ± 0.7c 

18 ± 0.7 

19 ± 1.2 

17 ± 1.3 

20 ± 1.2 

15 ± 1.0 

17 ± 1.8 

24 ± 2.3 

19 ± 1.5 

20 ± 2.4 

15 ± 0.7 

16 ± 1.9 

12 ± 1.5c 

37 ± 4.1 

45 ± 4.3 

26 ± 2.6c 

29 ± 6.0c 

54 ± 4.0c 

52 ± 2.2c 

31 ± 2.6 

27 ± 3.4 

33 ± 2.1 

29 ± 2.4 

31 ± 2.5 

46 ± 4.7 

34 ± 0.0 

30 ± 3.5 

37 ± 4.4 

42 ± 2.8 

49 ± 7.5 

36 ± 2.3c 

Trial summary 

Positive control 

Negative 

751 ± 2.0 

Negative 

569 ± 20.3 

Negative 

595 ± 29.0 

Negative 

594 ± 51.4 

Negative 

782 ± 20.3 

Negative 

468 ± 30.7 

+rat S9 

5% 10% 10% 10% 10% 30% 30% 

TA98 0 

(continued) 1 

3 

6 

10 

33 

100 

166 

333 

1,000 

3,333 

10,000 

28 ± 5.3 

42 ± 1.7 

41 ± 4.0c 

32 ± 3.7c 

30 ± 1.8c 

34 ± 3.8c 

28 ± 4.7 

56 ± 4.7 

26 ± 3.3c 

26 ± 5.8c 

42 ± 4.6c 

53 ± 2.4c 

22 ± 3.1 

27 ± 1.5 

29 ± 2.7 

36 ± 7.1 

37 ± 2.1 

28 ± 7.0c 

30 ± 2.0 

37 ± 4.2 

34 ± 2.1c 

37 ± 3.9c 

39 ± 1.7c 

44 ± 4.4c 

26 ± 1.9 

28 ± 3.8 

28 ± 5.1 

29 ± 1.9 

30 ± 3.3 

43 ± 2.9 

29 ± 4.7 

38 ± 4.1 

32 ± 5.8c 

29 ± 6.7c 

36 ± 4.1c 

38 ± 4.9c 

23 ± 5.0 

27 ± 4.8 

27 ± 6.1 

28 ± 4.6 

38 ± 2.3 

26 ± 1.5c 

Trial summary 

Positive control 

Negative 

681 ± 33.5 

Equivocal 

335 ± 6.6 

Equivocal 

293 ± 6.1 

Negative 

394 ± 5.6 

Negative 

514 ± 3.0 

Negative 

62 ± 4.5 

Negative 

135 ± 12.1 
148 D&C Yellow No. 11, NTP TR 463 

TABLE C1
 

Mutagenicity of D&C Yellow No. 11 in Salmonella typhimurium (continued)
 

Revertants/plate 

Strain Dose S9 +hamster S9 

(μg/plate) 30% 30% 

Study performed at Microbiological Associates, Inc. 

TA100 0 87 ± 4.2 108 ± 4.6 97 ± 2.3 

3.3 97 ± 4.7 

10 97 ± 3.0 106 ± 0.9 111 ± 3.5 

33 92 ± 7.5 141 ± 2.3 125 ± 6.4 

100 90 ± 5.3 151 ± 7.1 181 ± 7.1 

333 74 ± 3.5 154 ± 6.1 186 ± 11.0c 

1,000 200 ± 40.5c 

c c c 3,333 89 ± 10.2 154 ± 3.5 195 ± 11.0 

4,000 190 ± 4.7c 

Trial summary Negative Equivocal Positive 

Positive control 560 ± 10.1 531 ± 14.2 461 ± 45.5 

+rat S9 

5% 10% 30% 30% 30% 

TA100 

(continued) 

0 

3.3 

10 

33 

100 

333 

109 ± 6.0 

122 ± 6.4 

112 ± 4.0 

110 ± 6.9 

129 ± 8.9 

119 ± 2.8 

112 ± 8.1 

109 ± 10.0 

123 ± 4.4 

171 ± 5.7 

162 ± 10.1 

116 ± 4.5 

184 ± 11.1 

171 ± 7.5 

97 ± 3.2 

83 ± 1.7 

92 ± 4.6 

115 ± 13.6 

199 ± 1.8 

211 ± 13.4c 

1,000 

3,333 

4,000 

117 ± 2.8 

113 ± 4.2 

120 ± 6.0 

128 ± 9.9 

126 ± 4.2 

142 ± 12.6 

179 ± 7.7c 159 ± 4.1 

168 ± 1.2 

155 ± 9.8 

217 ± 20.0c 

217 ± 11.6c 

211 ± 2.9c 

Trial summary 

Positive control 

Negative 

530 ± 57.3 

Negative 

280 ± 16.8 

Weakly 

positive 

331 ± 0.0 

Equivocal 

411 ± 45.8 

Positive 

438 ± 15.8 

+ S9 

30% hamster 30% rat 

TA1538 0 

3.3 

10 

33 

100 

333 

1,000 

3,333 

4,000 

16 ± 2.3 

15 ± 2.0 

18 ± 2.1 

24 ± 2.1 

28 ± 0.9 

30 ± 2.2c 

30 ± 4.2c 

37 ± 2.9c 

29 ± 3.5c 

16 ± 2.2 

16 ± 3.8 

16 ± 0.3 

16 ± 5.2 

28 ± 1.5 

32 ± 2.7c 

23 ± 3.1c 

24 ± 1.5c 

28 ± 2.2c 

Trial summary 

Positive control 

Weakly 

positive 

108 ± 8.0 

Equivocal 

97 ± 3.1 
149 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE C1
 

Mutagenicity of D&C Yellow No. 11 in Salmonella typhimurium (continued)
 

Revertants/plate 

Strain Dose S9 +hamster S9 

(μg/plate) 5% 10% 30% 30% 30% 30% 

Study performed at Microbiological Associates, Inc.(continued) 

TA98 0 15 ± 3.5 18 ± 0.5 24 ± 3.1 30 ± 1.9 23 ± 1.5 22 ± 2.0 29 ± 1.2 

3.3 31 ± 3.2 

10 15 ± 3.7 32 ± 4.0 30 ± 2.1 

33 14 ± 2.0 35 ± 1.2 34 ± 3.8 

100 13 ± 3.2 26 ± 3.6 44 ± 2.0 51 ± 5.9 65 ± 3.8 39 ± 3.5 59 ± 2.6 

333 14 ± 1.5 25 ± 2.3 39 ± 3.5 46 ± 3.2 52 ± 7.5 52 ± 4.9 49 ± 3.3c 

c c 1,000 27 ± 4.8 40 ± 3.5 62 ± 1.7 53 ± 4.1 64 ± 5.0 c c c c 3,333 17 ± 2.7 28 ± 1.8 44 ± 3.1 47 ± 2.3 63 ± 2.0 52 ± 5.0 66 ± 3.5 c c 4,000 31 ± 2.0 43 ± 0.7 54 ± 3.6 52 ± 8.5 65 ± 1.2 

Trial summary Weakly 

Negative Negative Equivocal Negative positive Positive Positive 

Positive control 254 ± 3.8 93 ± 24.2 101 ± 20.9 75 ± 1.2 56 ± 2.0 53 ± 5.2 98 ± 4.1 

+rat S9 

5% 10% 30% 30% 30% 30% 

TA98 0 25 ± 5.7 30 ± 4.8 27 ± 3.4 21 ± 1.7 31 ± 3.0 27 ± 2.3 

(continued) 3.3 23 ± 3.8 

10 22 ± 4.9 26 ± 2.7 

33 34 ± 2.5 28 ± 4.0 

100 36 ± 4.1 44 ± 3.6 39 ± 0.3 36 ± 2.1 39 ± 1.5 48 ± 6.4 

333 23 ± 0.6 46 ± 1.2 40 ± 2.2 34 ± 4.4 37 ± 5.5 45 ± 2.7c 

c c 1,000 29 ± 3.0 39 ± 0.9 39 ± 5.4 50 ± 5.0 47 ± 2.5 c c c 3,333 38 ± 5.9 43 ± 3.3 45 ± 3.6 42 ± 4.1 55 ± 2.2 45 ± 2.3 c c 4,000 31 ± 4.6 39 ± 3.9 45 ± 1.5 61 ± 4.1 46 ± 0.9 

Trial summary Weakly Weakly 

Negative Negative Negative positive positive Negative 

Positive control 90 ± 4.4 78 ± 1.3 104 ± 3.6 92 ± 4.1 127 ± 11.5 158 ± 5.5 

a The detailed protocol and these data are presented in Zeigeret al. (1988); 0 μg/plate dose is the solvent control. b Revertants are presented as mean ± standard error from three plates.
 

Precipitate on plate d The positive controls in the absence of metabolic activation were sodium azide (TA100 and TA1535), 9-aminoacridine (TA97), and 

4-nitro-o-phenylenediamine (TA98 and TA1538). The positive control for metabolic activation with all strains was 2-aminoanthracene. 
150 D&C Yellow No. 11, NTP TR 463 

TABLE C2 

Induction of Sister Chromatid Exchanges in Chinese Hamster Ovary Cells by D&C Yellow No. 11 a 

Compound 

Dose 

(μg/mL) 

Total 

Cells 

No. of 

Chromosomes No. of 

SCEs 

SCEs/ 

Chromosome SCEs/ 

Cell 

Hrs 

in BrdU 

Relative 

Change of SCEs/ 

Chromosomeb 

(%) 

S9 

Trial 1 

Summary: Positive 

Dimethylsulfoxide 50 1,017 482 0.47 9.6 26.0 

Mitomycin-C 0.001 

0.010 

50 

5 

1,024 

102 

623 

216 

0.60 

2.11 

12.5 

43.2 

26.0 

26.0 

28.37 

346.82 

D&C Yellow No. 11 0.27 

0.80 

2.70 

8.00 

50 

50 

50 

0 

1,029 

1,015 

1,000 

579 

651 

871 

0.56 

0.64 

0.87 

11.6 

13.0 

17.4 

26.0 

26.0 

34.5c 

26.0 

18.72 

35.33* 

83.78* 

P<0.001d 

Trial 2 

Summary: Positive 

Dimethylsulfoxide 50 1,030 507 0.49 10.1 26.0 

Mitomycin-C 0.001 

0.010 

50 

5 

1,014 

104 

629 

191 

0.62 

1.83 

12.6 

38.2 

26.0 

26.0 

26.02 

273.10 

D&C Yellow No. 11 1.0 

1.5 

2.7 

5.0 

50 

50 

25 

0 

1,016 

1,019 

504 

900 

844 

522 

0.88 

0.82 

1.03 

18.0 

16.9 

20.9 

26.0 

33.0c 

33.0c 

26.0 

79.96* 

68.27* 

110.41* 

P<0.001 

+S9 

Summary: Positive 

Dimethylsulfoxide 50 1,022 457 0.44 9.1 26.0 

Cyclophosphamide 0.4 

2.0 

50 

5 

1,010 

103 

738 

245 

0.73 

2.37 

14.8 

49.0 

26.0 

26.0 

63.41 

431.94 

D&C Yellow No. 11 2.7 

8.0 

27.0e 

80.0e 

50 

50 

50 

0 

1,012 

1,025 

1,028 

532 

631 

1,055 

0.52 

0.61 

1.02 

10.6 

12.6 

21.1 

26.0 

26.0 

34.5c 

26.0 

17.56 

37.67* 

129.51* 

P<0.001 

* Positive response (P<0.01) a Study performed at Litton Bionetics, Inc. A detailed description of the protocol is presented in Gallowayet al. (1987). SCE=sister chromatid 

exchange; BrdU=bromodeoxyuridine b SCEs/chromosome in treated cells versus SCEs/chromosome in solvent control cells 

Because D&C Yellow No. 11 induced a delay in the cell division cycle, harvest time was extended to maximize the number of second- division 

metaphase cells available for analysis. d Significance of relative SCEs/chromosome tested by the linear regression trend test versus log of the dose e Color change (yellow); pH=7 

c 
151 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE C3 

Induction of Chromosomal Aberrations in Chinese Hamster Ovary Cells by D&C Yellow No. 11 a 

S9 +S9 

Dose Total No. of Abs/ Cells with Dose Total No. of Abs/ Cells with 

(μg/mL) Cells Abs Cell Abs (%) (μg/mL) Cells Abs Cell Abs (%) 

Trial 1 - Harvest time: 10.5 hours Harvest time: 21.5 hoursb 

Summary: Positive Summary: Positive 

Dimethylsulfoxide Dimethylsulfoxide 

200 3 0.02 1.0 200 1 0.01 0.5 

Mitomycin-C Cyclophosphamide 

0.15 200 32 0.16 9.5 6.25 200 32 0.16 12.0 

0.50 25 10 0.40 28.0 12.50 25 44 1.76 68.0 

D&C Yellow No. 11 D&C Yellow No. 11 

5.0 200 37 0.19 16.0* 60.0 25 92 3.68 88.0* 

7.5 50c 12 0.24 20.0* 69.7 25 109 4.36 80.0* 

10.0 100c 18 0.18 13.0* 80.0 25 86 3.44 76.0* 

15.0 0 

d P 0.001 P 0.001 

Trial 2 - Harvest time: 10.5 hours 

Summary: Positive 

Dimethylsulfoxide 

200 4 0.02 1.5 

Mitomycin-C 

0.15 200 24 0.12 9.5 

0.50 25 9 0.36 24.0 

D&C Yellow No. 11 

10.0 25 16 0.64 40.0* 

12.5 25 18 0.72 40.0* 

15.0 50 28 0.56 40.0* 

P 0.001 

* Positive (P 0.05) a Study performed at Litton Bionetics, Inc. The detailed description of the protocol is presented in Gallowayet al. (1987). Abs=aberrations b Because of significant chemical-induced cell cycle delay, incubation time prior to addition of Colcemid was lengthened to provide sufficient firstdivision metaphase cells at harvest. 

Less than 200 cells scored due to lack of readable cells 

d Significance of percent cells with aberrations tested by the linear regression trend test versus log of the dose 
152 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE C4 

Frequency of Micronuclei in Mouse Peripheral Blood Erythrocytes Following Treatment 

with D&C Yellow No. 11 in Feed for 13 Weeks a 

Dose 

(ppm) 

Micronucleated Normochromatic 

Erythrocytes/1,000 Cellsb 

Number of Mice 

per Dose Group 

Male 

0 

5,000 

17,000 

50,000 

1.73 ± 0.12 

1.97 ± 0.17 

1.58 ± 0.13 

1.71 ± 0.15 

10 

10 

10 

10 

P=0.504c 

Female 

0 

5,000 

17,000 

50,000 

1.23 ± 0.18 

1.28 ± 0.11 

0.94 ± 0.12 

1.21 ± 0.12 

10 

9 

9 

10 

P=0.848 

a Study performed at USDA Western Regional Center, CA. Smears were prepared from peripheral blood samples obtained at the termination of a 

13-week toxicity study on D&C Yellow No. 11 (NTP, 1991a). b At least 10,000 normochromatic erythrocytes (NCEs) were scored per animal. Data are presented as mean ± standard error.
 

Significance of micronucleated NCEs determined by analysis of variance
 
153 

APPENDIX D
 

ORGAN WEIGHTS AND
 

ORGAN-WEIGHT-TO-BODY-WEIGHT RATIOS
 

TABLE D1 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats 

at the 12-Month Interim Evaluation in the 2-Year Feed Study 

of D&C Yellow No. 11 ............................................................. 154
 
154 D&C Yellow No. 11, NTP TR 463 

TABLE D1 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats at the 12-Month Interim Evaluation 

in the 2-Year Feed Study of D&C Yellow No. 11a 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Male 

n 10 9 9 6 

Necropsy body wt 471 ± 10 461 ± 10 446 ± 6 440 ± 13 

R. Kidney 

Absolute 1.655 ± 0.042 1.680 ± 0.046 1.589 ± 0.045 1.620 ± 0.065 

Relative 3.52 ± 0.05 3.65 ± 0.06 3.56 ± 0.08 3.68 ± 0.07 

Liver 

Absolute 16.916 ± 0.599 19.417 ± 0.748** 19.599 ± 0.457** 20.480 ± 0.750** 

Relative 35.89 ± 0.75 42.03 ± 0.94** 43.95 ± 1.12** 46.49 ± 0.75** 

Female 

n 10 9 10 9 

Necropsy body wt 267 ± 8 262 ± 5 257 ± 2 252 ± 6 

R. Kidney 

Absolute 0.963 ± 0.021 0.912 ± 0.020 0.920 ± 0.027 0.930 ± 0.022 

Relative 3.62 ± 0.08 3.48 ± 0.05 3.58 ± 0.09 3.69 ± 0.04 

Liver 

Absolute 8.853 ± 0.204 9.656 ± 0.202* 10.566 ± 0.204** 10.780 ± 0.245** 

Relative 33.21 ± 0.58 36.87 ± 0.44** 41.06 ± 0.56** 42.82 ± 0.50** 

* Significantly different (P 0.05) from the control group by Williams’ or Dunnett’s test 

** P 0.01 a Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as mg 

organ weight/g body weight (mean ± standard error). 
155 

APPENDIX E
 

HEMATOLOGY RESULTS
 

TABLE E1	 Hematology Data for Rats at the 12-Month Interim Evaluation 

in the 2-Year Feed Study of D&C Yellow No. 11 ...................................... 156 
156 D&C Yellow No. 11, NTP TR 463 

TABLE E1 

Hematology Data for Rats at the 12-Month Interim Evaluation in the 2-Year Feed Study 

of D&C Yellow No. 11a 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Male 

n 10 9 9 6 

Hematocrit (%) 45.4 ± 0.4 43.6 ± 0.6* 43.4 ± 0.5** 43.0 ± 0.6** 

Hemoglobin (g/dL) 

Erythrocytes (10 /μL) 6 

Reticulocytes (10 /μL) 6 

Nucleated erythrocytes (10 /μL) 3 

15.3 ± 0.2 

8.76 ± 0.09 

0.22 ± 0.02 

0.06 ± 0.04 

14.7 ± 0.2* 

8.35 ± 0.09** 

0.24 ± 0.03 

0.08 ± 0.02 

14.3 ± 0.2** 

8.22 ± 0.09** 

0.28 ± 0.02 

0.01 ± 0.01 

14.5 ± 0.3** 

8.30 ± 0.16** 

0.23 ± 0.03 

0.05 ± 0.03 

Mean cell volume (fL) 51.9 ± 0.3 52.2 ± 0.3 52.8 ± 0.2 51.9 ± 0.4 

Mean cell hemoglobin (pg) 17.4 ± 0.1 17.6 ± 0.1 17.4 ± 0.1 17.4 ± 0.1 

Mean cell hemoglobin 

concentration (g/dL) 

Platelets (10 /μL) 3 

Leukocytes (10 /μL) 3 

Segmented neutrophils (10 /μL) 3 

Lymphocytes (10 /μL) 3 

Monocytes (10 /μL) 3 

Eosinophils (10 /μL) 3 

33.6 ± 0.3 

803.5 ± 20.7 

10.53 ± 1.09 

3.20 ± 0.61 

6.78 ± 0.52 

0.38 ± 0.08 

0.16 ± 0.04 

33.7 ± 0.2 

889.6 ± 16.0* 

8.63 ± 0.82 

2.66 ± 0.53 

5.66 ± 0.36 

0.19 ± 0.06 

0.12 ± 0.04 

33.0 ± 0.1 

897.9 ± 14.8** 

10.25 ± 0.55 

2.88 ± 0.22 

6.76 ± 0.53 

0.47 ± 0.08 

0.13 ± 0.03 

33.6 ± 0.4 

885.7 ± 54.1* 

8.89 ± 1.01 

2.55 ± 0.52 

6.01 ± 0.51 

0.24 ± 0.08 

0.09 ± 0.03 

Female 

n 10 9 10 9 

Hematocrit (%) 45.5 ± 0.4 45.6 ± 0.5 44.9 ± 0.4 44.7 ± 0.5 

Hemoglobin (g/dL) 

Erythrocytes (10 /μL) 6 

Reticulocytes (10 /μL) 6 

Nucleated erythrocytes (10 /μL) 3 

15.0 ± 0.1 

7.97 ± 0.07 

0.19 ± 0.02 

0.10 ± 0.04 

15.0 ± 0.1 

8.06 ± 0.06 

0.18 ± 0.02 

0.09 ± 0.04 

14.8 ± 0.1 

7.90 ± 0.07 

0.19 ± 0.02 

0.08 ± 0.03b 

14.7 ± 0.2 

7.86 ± 0.15 

0.21 ± 0.03 

0.05 ± 0.02 

Mean cell volume (fL) 57.1 ± 0.4 56.5 ± 0.4 56.9 ± 0.3 57.0 ± 0.9 

Mean cell hemoglobin (pg) 18.8 ± 0.1 18.7 ± 0.1 18.7 ± 0.1 18.7 ± 0.1 

Mean cell hemoglobin 

concentration (g/dL) 

Platelets (10 /μL) 3 

Leukocytes (10 /μL) 3 

Segmented neutrophils (10 /μL) 3 

Lymphocytes (10 /μL) 3 

Monocytes (10 /μL) 3 

Eosinophils (10 /μL) 3 

32.9 ± 0.2 

795.5 ± 67.3 

6.41 ± 0.69 

1.31 ± 0.24 

4.79 ± 0.50 

0.23 ± 0.07 

0.09 ± 0.02 

33.0 ± 0.2 

767.7 ± 11.0 

5.76 ± 0.60 

1.13 ± 0.16 

4.43 ± 0.49 

0.16 ± 0.04 

0.04 ± 0.01 

32.8 ± 0.1 

835.3 ± 65.1 

4.79 ± 0.19b 

0.79 ± 0.08b 

3.81 ± 0.19b 

0.17 ± 0.04b 

0.03 ± 0.02b 

32.8 ± 0.3 

794.7 ± 30.1 

5.41 ± 0.55 

1.12 ± 0.20 

4.04 ± 0.38 

0.21 ± 0.07 

0.04 ± 0.02 

* Significantly different (P 0.05) from the control group by Dunn’s or Shirley’s test 

** P 0.01 a Mean ± standard error. Statistical tests were performed on unrounded data. b n=9 
157 

APPENDIX F
 

REPRODUCTIVE TOXICITY STUDY RESULTS
 

TABLE F1 Body Weight Gains in F Rats in the Reproductive Toxicity Study 0 

of D&C Yellow No. 11 ............................................................. 158
 

TABLE F2 Precohabitation Feed Consumption by F Rats in the Reproductive Toxicity Study 0 

of D&C Yellow No. 11 ............................................................. 159
 

TABLE F3 Maternal Toxicity in F Rats in the Reproductive Toxicity Study 0 

of D&C Yellow No. 11 ............................................................. 160
 

TABLE F4 Developmental Toxicity in F1
 Rats in the Reproductive Toxicity Study

of D&C Yellow No. 11 ............................................................. 161
 
158 D&C Yellow No. 11, NTP TR 463 

TABLE F1 a Body Weight Gains in F0 Rats in the Reproductive Toxicity Study of D&C Yellow No. 11

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

n 60 60 60 60 

Male 

Days 

1 to 10 55.4 ± 0.5 52.7 ± 0.6** 50.6 ± 0.7** 48.8 ± 0.7** 

10 to 16 23.6 ± 0.7 24.6 ± 0.7 20.1 ± 0.9** 21.2 ± 0.7** 

16 to 22 31.1 ± 0.6 29.3 ± 0.5 29.2 ± 0.6 29.8 ± 0.5 

22 to 29 21.5 ± 0.7 16.2 ± 2.3** 24.0 ± 0.5 21.7 ± 0.5 

29 to 36 12.1 ± 0.5 16.7 ± 1.7** 13.9 ± 0.6 11.7 ± 0.5 

36 to 43 19.3 ± 0.5 15.7 ± 0.6** 13.7 ± 0.6** 15.2 ± 0.4** 

43 to 50 18.9 ± 0.4 18.2 ± 0.4 17.9 ± 0.4 15.9 ± 0.4** 

50 to 57 9.7 ± 0.6 12.6 ± 0.7** 12.0 ± 0.4* 10.6 ± 0.4 

57 to 64 8.0 ± 0.6 6.3 ± 0.7 6.8 ± 0.5 3.4 ± 1.7** 

64 to 71 9.7 ± 0.4 6.7 ± 0.5 7.2 ± 0.4 12.3 ± 2.4 

78 to 85 7.2 ± 0.6 10.7 ± 2.3 6.4 ± 0.5 6.7 ± 1.2 

1 to 71 209.3 ± 1.7 199.0 ± 1.6** 195.3 ± 1.6** 190.5 ± 2.0** 

1 to 85 228.6 ± 2.0 218.1 ± 2.0** 213.0 ± 1.8** 206.0 ± 1.7** 

Female 

Days 

1 to 3 10.1 ± 0.3b 9.7 ± 0.5 8.8 ± 0.2** 8.1 ± 0.2** 

3 to 11 20.4 ± 0.5 20.1 ± 0.5 20.3 ± 0.4 21.0 ± 0.4 

11 to 17 10.8 ± 0.4 10.6 ± 0.4 10.8 ± 0.4 9.6 ± 0.4 

17 to 24 7.9 ± 0.3 7.1 ± 0.4 7.5 ± 0.3 7.2 ± 0.3 

24 to 31 9.9 ± 0.4 9.7 ± 0.3 9.1 ± 0.3 10.1 ± 0.4 

31 to 38 7.7 ± 0.4 7.5 ± 0.3 7.5 ± 0.4 6.2 ± 0.3* 

38 to 45 5.1 ± 0.4 4.6 ± 0.3 4.7 ± 0.4 4.9 ± 0.5 

45 to 52 6.5 ± 0.4 6.3 ± 0.3 5.5 ± 0.3* 5.2 ± 0.4** 

52 to 59 2.0 ± 0.3 3.0 ± 0.4 3.8 ± 0.4** 3.1 ± 0.3** 

59 to 66 5.2 ± 0.4 5.9 ± 0.4 5.6 ± 0.4 5.3 ± 0.3 

1 to 66 85.6 ± 1.1b 84.3 ± 0.9 83.4 ± 1.1 80.5 ± 1.0** 

* Significantly different (P 0.05) from the control group by Williams’ or Dunnett’s test 

** P 0.01 a Body weight gains are given in grams (mean ± standard error). b n=59 
159 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE F2 a Precohabitation Feed Consumption by F0 Rats in the Reproductive Toxicity Study of D&C Yellow No. 11

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Number of cagesb 12 12 12 12 

Male 

Days 

2 to 9 

16 to 23 

23 to 30 

30 to 37 

37 to 44 

44 to 51 

51 to 58 

58 to 65 

17.0 ± 0.2 

17.0 ± 0.1 

18.0 ± 0.2 

18.4 ± 0.5 

18.7 ± 0.8 

18.8 ± 0.8 

15.8 ± 1.6 

16.1 ± 0.3 

16.3 ± 0.5 

17.0 ± 0.1 

17.0 ± 0.6 

18.0 ± 0.3 

18.4 ± 0.2 

23.8 ± 0.6** 

10.4 ± 1.9 

16.4 ± 0.5 

15.8 ± 0.3** 

16.4 ± 0.1** 

17.2 ± 0.7* 

17.3 ± 0.9 

18.3 ± 0.1 

23.4 ± 0.5* 

11.2 ± 2.0 

16.8 ± 0.5 

16.2 ± 0.4 

16.8 ± 0.1 

17.3 ± 0.4 

16.8 ± 0.4* 

17.9 ± 0.1 

21.6 ± 1.1 

11.8 ± 2.0c 

17.0 ± 0.6 

Female 

Days 

2 to 9 

9 to 16 

16 to 23 

23 to 30 

30 to 37 

37 to 44 

44 to 51 

51 to 58 

58 to 65 

12.6 ± 0.1 

11.4 ± 0.1 

11.1 ± 0.1 

11.4 ± 0.7 

11.6 ± 0.2 

11.7 ± 1.0 

10.7 ± 1.0 

12.0 ± 0.6 

10.5 ± 0.3 

12.1 ± 0.1* 

11.1 ± 0.1 

11.1 ± 0.1 

12.2 ± 0.9 

11.2 ± 0.1 

11.9 ± 0.2 

10.3 ± 0.4 

11.8 ± 0.8 

10.0 ± 0.4 

12.1 ± 0.2* 

11.1 ± 0.1 

10.9 ± 0.1 

10.7 ± 0.3* 

11.2 ± 0.1 

12.0 ± 0.1 

10.3 ± 0.5 

11.4 ± 0.2 

10.4 ± 0.1 

12.3 ± 0.1 

11.2 ± 0.1 

11.0 ± 0.2 

11.3 ± 0.1 

11.4 ± 0.2 

11.7 ± 0.1 

10.9 ± 0.1 

11.4 ± 0.3 

10.8 ± 0.3 

* Significantly different (P 0.05) from the control group by Dunn’s or Shirley’s test 

** P 0.01 a Feed consumption data are given in grams per animal per day (mean ± standard error). b Five rats per cage
 

Feed consumption was not measured for one cage.
 
160 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE F3 

Maternal Toxicity in F Rats in the Reproductive Toxicity Study of D&C Yellow No. 11 a 0 

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Number examined 60 60 60 60 

Number pregnant 43 39 49 46 

Maternal body weight gains during gestation (g) 

Days 

0 to 6 9.1 ± 2.7 10.9 ± 4.0 10.9 ± 3.0 10.7 ± 3.7 

6 to 15 

15 to 21 

32.9 ± 4.1 

47.0 ± 9.3 

31.2 ± 5.1 

50.0 ± 7.5 

28.8 ± 4.9** 

47.6 ± 13.0 

30.4 ± 4.5* 

49.1 ± 7.3b 

0 to 21 89.0 ± 10.8 92.2 ± 12.2 87.4 ± 17.0 90.4 ± 10.9b 

Maternal body weight gains during lactation (g) 

Days 

1 to 4 

4 to 14 

14 to 21 

–3.5 ± 6.8 

18.6 ± 12.9 

–0.2 ± 16.2 

–5.8 ± 8.0c 

13.9 ± 14.0c 

1.9 ± 18.9 

–7.7 ± 11.8d 

18.1 ± 11.1d 

–1.6 ± 16.0f 

–8.0 ± 9.4e 

15.5 ± 9.4e 

2.4 ± 16.8e 

1 to 21 14.9 ± 9.4 10.3 ± 13.6 8.9 ± 15.6g 9.9 ± 15.0e 

Duration of gestation (days)h 23.0 ± 0.0 23.0 ± 0.2 23.1 ± 0.3 23.0 ± 0.0 

* Significantly different (P 0.05) from the control group by Dunnett’s test 

** P 0.01 a Mean ± standard deviation 

b n=44
 

n=38
 d n=46 e n=45 

f n=48 

g n=47 

h Data for rats with confirmed mating dates 
161 

c 

D&C Yellow No. 11, NTP TR 463 

TABLE F4 

Developmental Toxicity in F a 1 Rats in the Reproductive Toxicity Study of D&C Yellow No. 11

0 ppm 500 ppm 1,700 ppm 5,000 ppm 

Number of litters examined 43 39 49 46 

Pups delivered (total) 418 396 471 459 

Pups delivered per litter 9.7 ± 2.5 10.2 ± 2.3 9.6 ± 3.3 10.0 ± 2.0 

Percent male pups 50.0 ± 16.9 51.8 ± 18.5 48.2 ± 18.1 48.7 ± 20.2 

Pups surviving 4 days (precull)

 per number of pups delivered 410/416 (99%) 388/396 (98%) 463/470 (99%) 447/458 (98%) 

Pups surviving 21 days per number of pups

 selected on day 4 (postcull) 324/327 (99%) 297/299 (99%) 349/349 (100%) 347/350 (99%) 

Pup weight per litter (g) 

Day 

1 

4 (precull) 

4 (postcull) 

14 

21 

5.23 ± 0.04 

7.26 ± 0.08c 

7.29 ± 0.08c 

20.7 ± 0.2 

30.8 ± 0.3 

5.33 ± 0.05 

7.22 ± 0.12d 

7.28 ± 0.12d 

19.6 ± 0.2** 

28.8 ± 0.3** 

5.37 ± 0.04*b 

7.13 ± 0.13e 

7.18 ± 0.12e 

19.5 ± 0.2**b 

28.1 ± 0.2**b 

5.34 ± 0.03 

6.96 ± 0.07f 

7.01 ± 0.06f 

19.1 ± 0.2**g 

27.1 ± 0.2**g 

* Significantly different (P 0.05) from the control group by Williams’ or Dunnett’s test 

** P 0.01 a Data are presented as mean ± standard deviation (pups delivered/litter) or mean ± standard error (pup weights/litter). b n=48
 

n=21
 d n=23 e n=25 

f n=28 

g n=45 
162 D&C Yellow No. 11, NTP TR 463 
163 

APPENDIX G
 

CHEMICAL CHARACTERIZATION AND
 

DOSE FORMULATION STUDIES
 

PROCUREMENT AND CHARACTERIZATION OFD&C YELLOW NO. 11 .................................... 164
 

PREPARATION AND ANALYSIS OF DOSE FORMULATIONS .............................................. 165
 

FIGURE G1 Infrared Absorption Spectrum of D&C Yellow No. 11 ................................. 166
 

FIGURE G2 Nuclear Magnetic Resonance Spectrum of D&C Yellow No. 11 .......................... 167
 

TABLE G1 Preparation and Storage of Dose Formulations in the Feed Studies
 

of D&C Yellow No. 11 ............................................................. 168
 

TABLE G2 Results of Analyses of Dose Formulations Administered to Rats
 

in the Reproductive Toxicity and 2-Year Feed Studies of D&C Yellow No. 11 ............. 169
 

TABLE G3 Results of Referee Analysis of Dose Formulations Administered to Rats
 

in the Reproductive Toxicity and 2-Year Feed Studies of D&C Yellow No. 11 ............. 174
 
164 D&C Yellow No. 11, NTP TR 463 

CHEMICAL CHARACTERIZATION AND 

DOSE FORMULATION STUDIES 

PROCUREMENT AND CHARACTERIZATION OF D&C YELLOW NO. 11 

D&C Yellow No. 11 was obtained from H. Kohnstamm and Company, Inc. (New York), in one lot (ZB2016) 

and certified by the Food and Drug Administration, Division of Color Technology. Lot ZB2016 was used during 

the reproductive toxicity and 2-year studies. Identity, purity, and stability analyses were conducted by the 

analytical chemistry laboratory, Midwest Research Institute (Kansas City, MO). Reports on analyses performed 

in support of the D&C Yellow No. 11 studies are on file at the National Institute of Environmental Health 

Sciences. 

Lot ZB2016, a yellow powder, was identified as D&C Yellow No. 11 by infrared, ultraviolet/visible, and nuclear 

magnetic resonance spectrometry. All spectra were consistent with those expected for the structure of D&C 

Yellow No. 11. However, the nuclear magnetic resonance spectrum indicated impurities. Direct inlet mass 

spectrometry confirmed the identity of the compound as D&C Yellow No. 11 and indicated the presence of a 

monochlorinated isomer. The infrared and nuclear magnetic resonance spectra are presented in Figures G1 and 

G2. The observed melting point range, 240.9 to 242.1 C, was consistent with the melting point range, 235 to 

240 C, specified by the manufacturer for purified D&C Yellow No. 11. 

The purity was determined by elemental analyses, Karl Fischer water analysis, thin-layer chromatography (TLC), 

and high-performance liquid chromatography (HPLC). TLC was performed on Silica Gel 60 F-254 plates with 

two solvent systems: 1) methylene chloride:acetone:glacial acetic acid (70:26:4) and 2) toluene:methanol (95:5). 

Quinoline was used as a reference standard. Plates were examined under visible and ultraviolet light (254 and 

366 nm) and with iodine vapors. HPLC was performed with a Waters μBondapak C 18 column using ultraviolet 

(280 nm) and visible (436 nm) detection and a solvent system of water:methanol (37:63) at a flow rate of 1.0 

mL/min. 

Elemental analyses for carbon, hydrogen, and nitrogen were in agreement with the theoretical values for D&C 

Yellow No. 11. Karl Fischer water analysis indicated less than 0.02% water. TLC indicated one major spot by 

system 1 and one major spot and one trace impurity by system 2. HPLC indicated a major peak and two 

impurities with areas greater than 0.1% relative to the major peak at both 280 and 436 nm. A high-speed 

scanning detector (Hewlett-Packard 1040A) was used in conjunction with HPLC with a solvent system of 

water:methanol (32:68) to obtain an ultraviolet/visible absorption spectrum for the largest of the two impurity 

peaks. The results indicated that this impurity was similar in structure to that of the major peak. The overall 

purity was determined to be approximately 99%. 

Stability studies of the bulk chemical were performed using the HPLC system described for the purity analysis 

except with a solvent system ratio of 32:68, ultraviolet detection at 254 nm, and valerophenone as an internal 

standard. These studies indicated that D&C Yellow No. 11 was stable as a bulk chemical for 2 weeks when 

stored protected from light at temperatures up to 60 C. To ensure stability, the bulk chemical was stored in its 

original packaging (metal cans or cardboard drums) at room temperature protected from light. The stability of 

the bulk chemical was monitored by the study laboratory approximately every 4 months during the studies and 

within 30 days of the end of the 2-year study by HPLC. No degradation of the bulk chemical was observed. 

PREPARATION AND ANALYSIS OF DOSE FORMULATIONS 

The dose formulations were prepared every 2 weeks by mixing D&C Yellow No. 11 with feed (Table G1). A 

D&C Yellow No. 11/feed premix was made by hand and then blended with feed in a Patterson-Kelly twin-shell 
D&C Yellow No. 11, NTP TR 463 165 

blender for 15 minutes with the intensifier bar on for the first 5 minutes. During the studies, dose formulations 

were stored in double-thickness plastic bags in rigid plastic containers at room temperature protected from light 

for up to 3 weeks. 

Homogeneity and stability studies of the 500 ppm dose formulation were performed by the analytical chemistry 

laboratory. Extracts were prepared by shaking 10 g samples with 100 mL of acetone in a wrist-action shaker for 

15 minutes. After centrifugation, 5 mL aliquots of the extracts were diluted to 50 mL with a water:methanol 

solution (20:80) and filtered. HPLC was performed with a Brownlee RP-18 column using visible light detection 

and a mobile phase of water:methanol (20:80) at a flow rate of 1.0 mL/minute. Homogeneity was confirmed, 

and the dose formulations were determined to be stable for up to 3 weeks when stored protected from light at 

room temperature and for 7 days when stored open to air and light. 

Periodic analyses of the dose formulations of D&C Yellow No. 11 were conducted at the study laboratory using 

visible spectrometry. Dose formulations were analyzed approximately every 8 to 10 weeks (Table G2). All dose 

formulations used in the studies were within 10% of the target concentrations. One formulation was remixed due 

to an unacceptable ratio of duplicate analyses. The remix was within acceptable limits and the original mix was 

not used for dosing. Results of a referee analysis performed by the analytical chemistry laboratory indicated 

good agreement with the results obtained by the study laboratory (Table G3). 
166 D&C Yellow No. 11, NTP TR 463 

FIGURE G1
 

Infrared Absorption Spectrum of D&C Yellow No. 11
 
D&C Yellow No. 11, NTP TR 463 167 

FIGURE G2
 

Nuclear Magnetic Resonance Spectrum of D&C Yellow No. 11
 
168 D&C Yellow No. 11, NTP TR 463 

TABLE G1 

Preparation and Storage of Dose Formulations in the Feed Studies of D&C Yellow No. 11 

Reproductive Toxicity Study 2-Year Study 

Preparation 

A premix of feed and D&C Yellow No. 11 was prepared, then layered 

into the remaining feed and blended in a Patterson-Kelly twin-shell 

blender with the intensifier bar on for 5 minutes and off for 10 minutes. 

Doses were prepared every 2 weeks. 

Same as reproductive toxicity study 

Chemical Lot Number 

ZB2016 ZB2016 

Maximum Storage Time 

3 weeks 3 weeks 

Storage Conditions 

Stored in double-thickness plastic bags in rigid plastic containers at 

room temperature in the dark. 

Same as reproductive toxicity study 

Study Laboratory 

Southern Research Institute 

(Birmingham, AL) 

Southern Research Institute 

(Birmingham, AL) 

Referee Laboratory 

Midwest Research Institute 

(Kansas City, MO) 

Midwest Research Institute 

(Kansas City, MO) 
D&C Yellow No. 11, NTP TR 463 169 

TABLE G2 

Results of Analyses of Dose Formulations Administered to Rats in the Reproductive Toxicity 

and 2-Year Feed Studies of D&C Yellow No. 11 

Target Determined Difference 

Date Prepared Date Analyzed Concentration Concentrationa from Target 

(ppm) (ppm) (%) 

27 November 1989b 27–28 November 1989 500 

500 

483c 

486d 

–3 

–3 

500 483e –3 

5,000 

5,000 

5,050c 

4,910d 

+1 

–2 

5,000 4,980e 0 

12 December 1989 13 December 1989 500 480 –4 

500 489 –2 

500 486 –3 

1,700 1,700 0 

1,700 1,680 –1 

1,700 1,700 0 

5,000 5,040 +1 

5,000 4,980 0 

5,000 5,030 +1 

20 February 1990 20–21 February 1990 500 475 –5 

500 472 –6 

500 484 –3 

500 481 –4 

1,700 1,680 –1 

1,700 1,680 –1 

1,700 1,710 +1 

1,700 1,680 –1 

5,000 4,990 0 

5,000 5,100 +2 

5,000 5,070 +1 

5,000 4,980 0 

17 April 1990 18 April 1990 500 473 –5 

500 454 –9 

500 470 –6 

500 478 –4 

1,700 1,630 –4 

1,700 1,600 –6 

1,700 1,580 –7 

1,700 1,640 –4 

5,000 4,780 –4 

5,000 4,880 –2 

5,000 4,780 –4 

5,000 4,880 –2 
170 D&C Yellow No. 11, NTP TR 463 

TABLE G2 

Results of Analyses of Dose Formulations Administered to Rats in the Reproductive Toxicity 

and 2-Year Feed Studies of D&C Yellow No. 11 (continued) 

Target Determined Difference 

Date Prepared Date Analyzed Concentration Concentration from Target 

(ppm) (ppm) (%) 

12 June 1990 13–14 June 1990 500 471 –6 

500 468 –6 

500 487 –3 

500 461 –8 

1,700 1,650 –3 

1,700 1,630 –4 

1,700 1,540 –9 

1,700 1,550 –9 

5,000 4,750 –5 

5,000 4,520 –10 

5,000 4,610 –8 

5,000 4,970 –1 

f 6 July 1990 500 431 –14 

1,700 1,620 –5 

5,000 4,760 –5 

21 August 1990 21 August 1990 500 503 +1 

500 494 –1 

500 519 +4 

500 500 0 

1,700 1,680 –1 

1,700 1,700 0 

1,700 1,680 –1 

1,700 1,670 –2 

5,000 4,910 –2 

5,000 4,930 –1 

5,000 4,970 –1 

5,000 5,060 +1 

16 October 1990 16–17 October 1990 500 501 0 

500 495 –1 

500 488 –2 

500 488 –2 

1,700 1,670 –2 

1,700 1,670 –2 

1,700 1,660 –2 

1,700 1,710 +1 

5,000 4,970 –1 

5,000 4,890 –2 

5,000 4,960 –1 

5,000 4,950 –1 
D&C Yellow No. 11, NTP TR 463 171 

TABLE G2 

Results of Analyses of Dose Formulations Administered to Rats in the Reproductive Toxicity 

and 2-Year Feed Studies of D&C Yellow No. 11 (continued) 

Target Determined Difference 

Date Prepared Date Analyzed Concentration Concentration from Target 

(ppm) (ppm) (%) 

11 December 1990 12 December 1990 500 491 –2 

500 494 –1 

500 494 –1 

500 500 0 

1,700 1,720 +1 

1,700 1,690 –1 

1,700 1,700 0 

1,700 1,690 –1 

5,000 5,000 0 

5,000 5,060 +1 

5,000 5,020 0 

5,000 5,010 0 

f 7 January 1991 500 477 –5 

1,700 1,650 –3 

5,000 4,910 –2 

5 February 1991 5–7 February 1991 500 495 –1 

500 501 0 

500 495 –1 

500 526 +5 

1,700 1,670 –2 

1,700 1,810 +6 

1,700 1,680 –1 

1,700 1,670 –2 

5,000 4,940 –1 

5,000 4,840 –3 

5,000 4,900 –2 

5,000 4,840 –3 

2 April 1991 3–4 April 1991 500 469 –6 

500 466 –7 

500 473 –5 

500 469 –6 

1,700 1,700 0 

1,700 1,700 0 

1,700 1,720 +1 

1,700 1,710 +1 

5,000 5,130 +3 

5,000 5,310 +6 

5,000 5,100 +2 

5,000 5,110 +2 
172 D&C Yellow No. 11, NTP TR 463 

TABLE G2 

Results of Analyses of Dose Formulations Administered to Rats in the Reproductive Toxicity 

and 2-Year Feed Studies of D&C Yellow No. 11 (continued) 

Target Determined Difference 

Date Prepared Date Analyzed Concentration Concentration from Target 

(ppm) (ppm) (%) 

11 June 1991 11–12 June 1991 500 484 –3 

500 496 –1 

500 493 –1 

500 487 –3 

1,700 1,680 –1 

1,700 1,680 –1 

1,700 1,670 –2 

1,700 1,680 –1 

5,000 4,940 –1 

5,000 4,960 –1 

5,000 4,970 –1 

5,000 4,970 –1 

f 25 and 27 June 1991 500 461 –8 

500 480 –4 

500 471 –6 

500 464 –7 

1,700 1,640 –4 

1,700 1,640 –4 

1,700 1,630 –4 

1,700 1,660 –2 

5,000 4,860 –3 

5,000 4,860 –3 

5,000 4,940 –1 

5,000 4,930 –1 

20 August 1991 20–21 August 1991 500 505 +1 

500 512 +2 

500 509 +2 

500 509 +2 

1,700 1,710 +1 

1,700 1,700 0 

1,700 1,700 0 

1,700 1,700 0 

5,000 4,940 –1 

5,000 4,960 –1 

5,000 5,080 +2 

5,000 4,970 –1 

15 October 1991 16–17 October 1991 500 479 –4 

500 482 –4 

500 488 –2 

500 498 0 

1,700 1,680 –1 

1,700 1,680 –1 

1,700 1,650 –3 

1,700 1,680 –1 

5,000 4,860 –3 

5,000 4,850 –3 

5,000 4,940 –1 

5,000 4,910 –2 
D&C Yellow No. 11, NTP TR 463 173 

TABLE G2 

Results of Analyses of Dose Formulations Administered to Rats in the Reproductive Toxicity 

and 2-Year Feed Studies of D&C Yellow No. 11 (continued) 

Target Determined Difference 

Date Prepared Date Analyzed Concentration Concentration from Target 

(ppm) (ppm) (%) 

10 December 1991 10–11 December 1991 500 493 –1 

500 502 0 

500 505 +1 

500 489 –2 

1,700 1,670 –2 

1,700 1,650 –3 

1,700 1,670 –2 

1,700 1,680 –1 

5,000 4,920 –2 

5,000 4,870 –3 

5,000 5,080 +2 

5,000 4,930 –1 

31 December 1991 – 

2–3 January 1992f 

500 

500 

480 

480 

–4

–4 

500 470 –6 

1,700 1,650 –3 

1,700 1,680 –1 

1,700 1,650 –3 

5,000 4,890 –2 

5,000 4,840 –3 

5,000 4,940 –1 

4 February 1992 4–5 February 1992 500 501 0 

500 517 +3 

500 501 0 

500 494 –1 

1,700 1,700 0 

1,700 1,780g +5 

1,700 1,690 –1 

1,700 1,690 –1 

5,000 4,940 –1 

5,000 4,930 –1 

5,000 5,020 0 

5,000 4,930 –1 

6 February 1992 7 February 1992 1,700 1,690h –1 
174 D&C Yellow No. 11, NTP TR 463 

TABLE G2 

Results of Analyses of Dose Formulations Administered to Rats in the Reproductive Toxicity 

and 2-Year Feed Studies of D&C Yellow No. 11 (continued) 

Target Determined Difference 

Date Prepared Date Analyzed Concentration Concentration from Target 

(ppm) (ppm) (%) 

31 March 1992 31 March 1992 – 500 493 –1

 1 April 1992 500 499 0 

500 502 0 

1,700 1,690 –1 

1,700 1,670 –2 

1,700 1,680 –1 

5,000 4,920 –2 

5,000 5,050 +1 

5,000 4,870 –3 

a Results of duplicate analyses b Homogeneity analyses, formulations not used for dosing c Sample from top right of twin-shell blender d Sample from top left of twin-shell blender e Sample from bottom of twin-shell blender f Animal room samples 

g Not used for dosing due to unacceptable ratio of duplicate analyses (0.89) h Result of remix 

TABLE G3 

Results of Referee Analysis of Dose Formulations Administered to Rats in the Reproductive Toxicity 

and 2-Year Feed Studies of D&C Yellow No. 11 

Determined Concentration (ppm) 

Date Prepared 

(ppm) 

Target Concentration 

Laboratorya Study 

Laboratoryb 

Referee 

12 December 1989 500 480 500 ± 2 

a Results of duplicate analyses b Results of triplicate analyses (mean ± standard error) 
175 

APPENDIX H 

FEED AND COMPOUND CONSUMPTION 

IN THE 2-YEAR FEED STUDY 

OF D&C YELLOW NO. 11 

TABLE H1 

TABLE H2 

Feed and Compound Consumption by Male Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 ............................................................ 

Feed and Compound Consumption by Female Rats in the 2-Year Feed Study 

of D&C Yellow No. 11 ............................................................ 

176 

177 
176 D&C Yellow No. 11, NTP TR 463 

TABLE H1 

Feed and Compound Consumption by Male Rats in the 2-Year Feed Study of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 

Feed Body Feed Body Dose/ Feed Body Dose/ 

(g/day) Weight (g/day) Weight Day (g/day) Weight Day a b 

Week (g) (g) (mg/kg) (g) (mg/kg) 

5,000 ppm 

Feed Body Dose/ 

(g/day) Weight Day 

(g) (mg/kg) 

2 15.1 159 14.3 145 50 13.9 142 167 

6 18.1 277 18.6 260 36 18.5 253 125 

10 18.1 327 17.9 315 28 18.0 310 99 

13 16.7 353 17.2 343 25 16.8 337 85 

17 17.2 383 16.7 371 23 17.2 365 80 

21 17.0 401 16.3 390 21 17.1 385 76 

25 16.1 416 16.5 405 20 15.7 400 67 

29 16.7 431 17.1 421 20 17.1 413 70 

33 14.9 425 15.0 427 18 15.9 423 64 

37 16.5 449 17.8 436 20 17.5 432 69 

41 16.9 455 17.0 443 19 17.2 439 66 

45 17.5 456 16.8 449 19 17.2 443 66 

49 16.4 464 16.9 454 19 17.0 450 64 

53 16.5 472 16.6 460 18 15.9 457 59 

57 17.1 472 16.6 461 18 16.8 460 62 

61 16.7 475 16.1 462 17 16.3 461 60 

65 15.9 477 15.7 463 17 15.9 460 59 

69 15.7 475 15.8 463 17 16.3 462 60 

73 16.1 472 15.2 460 17 16.4 456 61 

77 15.7 473 15.4 451 17 16.7 451 63 

81 16.1 470 15.4 446 17 15.8 442 61 

85 14.2 463 14.8 445 17 14.9 435 58 

89 14.3 452 14.0 439 16 15.6 433 61 

93 14.3 451 14.1 425 17 15.8 422 64 

97 12.7 446 14.9 425 17 13.3 420 54 

101 13.6 435 14.2 414 17 16.5 403 70 

13.5 133 506 

17.5 240 363 

18.2 297 308 

17.1 323 265 

16.9 352 240 

16.7 370 226 

16.7 384 217 

17.6 400 220 

16.7 404 206 

16.8 416 202 

17.4 421 207 

16.7 423 197 

16.9 430 197 

17.3 438 198 

17.3 438 197 

16.5 441 187 

16.3 439 185 

16.6 438 189 

16.5 436 189 

17.1 433 197 

16.1 425 190 

15.3 413 185 

15.2 404 188 

15.1 401 188 

13.6 387 175 

15.3 368 208 

Means for weeks 

1-13 17.0 279 17.0 265 35 16.8 260 119 

14-52 16.6 431 16.7 422 20 16.9 417 69 

53-101 15.3 464 15.3 447 17 15.9 443 61 

16.6 248 360 

16.9 400 212 

16.0 420 191 

Grams of feed consumed per animal per day a 

Milligrams of D&C Yellow No. 11 consumed per kilogram body weight per day b 
D&C Yellow No. 11, NTP TR 463 177 

TABLE H2 

Feed and Compound Consumption by Female Rats in the 2-Year Feed Study of D&C Yellow No. 11 

0 ppm 500 ppm 1,700 ppm 

Feed Body Feed Body Dose/ Feed Body Dose/ 

(g/day) Weight (g/day) Weight Day (g/day) Weight Day a b 

Week (g) (g) (mg/kg) (g) (mg/kg) 

5,000 ppm 

Feed Body Dose/ 

(g/day) Weight Day 

(g) (mg/kg) 

2 12.1 126 11.0 117 47 10.9 115 161 

6 12.1 168 12.1 161 38 11.3 156 122 

10 11.5 187 11.4 179 32 11.0 173 108 

16 10.7 206 10.7 200 27 10.2 196 88 

21 10.4 212 10.7 205 26 10.2 201 86 

24 10.4 217 10.7 212 25 10.2 207 84 

28 11.0 227 11.0 218 25 10.3 215 81 

32 10.2 233 10.9 227 24 10.5 221 80 

36 9.9 238 10.4 231 23 10.0 222 77 

40 11.2 246 11.4 240 24 11.7 231 86 

44 11.6 255 11.1 248 22 11.0 238 79 

48 12.3 265 12.2 258 24 11.9 251 80 

52 11.9 277 12.3 276 22 11.7 259 76 

56 12.5 290 12.2 285 21 12.1 273 75 

60 12.1 296 12.9 293 22 12.1 280 73 

64 12.8 307 12.1 303 20 12.1 289 71 

68 12.6 314 12.2 312 20 12.4 300 70 

72 12.4 320 12.7 317 20 12.6 304 70 

76 12.2 323 12.6 320 20 12.2 305 68 

80 12.9 331 12.6 323 20 12.6 310 69 

84 12.8 339 12.8 330 19 12.2 315 66 

88 12.0 342 12.1 330 18 12.5 320 66 

92 12.4 348 11.5 333 17 12.6 327 65 

96 13.2 357 13.0 342 19 12.6 331 65 

100 12.2 355 12.7 344 18 12.6 336 64 

104 11.6 354 12.4 349 18 12.2 337 61 

10.9 113 484 

11.6 155 376 

11.0 174 315 

10.5 194 272 

10.8 197 274 

10.8 205 263 

10.7 211 253 

11.0 219 253 

10.1 223 226 

12.3 233 264 

11.4 240 238 

12.1 250 241 

11.7 263 223 

12.2 275 221 

12.5 282 221 

11.7 291 201 

12.3 299 205 

13.4 307 219 

12.8 308 208 

12.6 313 201 

11.6 317 183 

12.5 319 196 

12.6 320 197 

12.2 332 184 

12.5 334 188 

12.2 333 183 

Means for weeks 

1-13 11.9 160 11.5 152 39 11.0 148 131 

14-52 11.0 238 11.1 231 24 10.8 224 82 

53-104 12.4 329 12.5 322 19 12.4 310 68 

11.2 147 392 

11.1 224 251 

12.4 310 201 

Grams of feed consumed per animal per day a 

Milligrams of D&C Yellow No. 11 consumed per kilogram body weight per day b 
178 D&C Yellow No. 11, NTP TR 463 
179 

APPENDIX I 

INGREDIENTS, NUTRIENT COMPOSITION, 

AND CONTAMINANT LEVELS 

IN NIH-07 RAT AND MOUSE RATION 

TABLE I1 

TABLE I2 

TABLE I3 

TABLE I4 

Ingredients of NIH-07 Rat and Mouse Ration .......................................... 

Vitamins and Minerals in NIH-07 Rat and Mouse Ration................................. 

Nutrient Composition of NIH-07 Rat and Mouse Ration .................................. 

Contaminant Levels in NIH-07 Rat and Mouse Ration ................................... 

180 

180 

181 

182 
180 D&C Yellow No. 11, NTP TR 463 

TABLE I1 a Ingredients of NIH-07 Rat and Mouse Ration

Ingredientsb Percent by Weight 

Ground #2 yellow shelled corn 24.50 

Ground hard winter wheat 23.00 

Soybean meal (49% protein) 12.00 

Fish meal (60% protein) 10.00 

Wheat middlings 10.00 

Dried skim milk 5.00 

Alfalfa meal (dehydrated, 17% protein) 4.00 

Corn gluten meal (60% protein) 3.00 

Soy oil 2.50 

Dried brewer’s yeast 2.00 

Dry molasses 1.50 

Dicalcium phosphate 1.25 

Ground limestone 0.50 

Salt 0.50 

Premixes (vitamin and mineral) 0.25 

a NCI, 1976; NIH, 1978 b Ingredients were ground to pass through a U.S. Standard Screen No. 16 before being mixed. 

TABLE I2 

Vitamins and Minerals in NIH-07 Rat and Mouse Ration a 

Amount Source 

Vitamins 

A 5,500,000 IU Stabilized vitamin A palmitate or acetate 

D3 4,600,000 IU D-activated animal sterol 

K3 2.8 g Menadione 

d­ -Tocopheryl acetate 20,000 IU 

Choline 560.0 g Choline chloride 

Folic acid 2.2 g 

Niacin 30.0 g 

d-Pantothenic acid 18.0 g d-Calcium pantothenate 

Riboflavin 3.4 g 

Thiamine 10.0 g Thiamine mononitrate 

B12 4,000 μg 

Pyridoxine 1.7 g Pyridoxine hydrochloride 

Biotin 140.0 mg d-Biotin 

Minerals 

Iron 120.0 g Iron sulfate 

Manganese 60.0 g Manganous oxide 

Zinc 16.0 g Zinc oxide 

Copper 4.0 g Copper sulfate 

Iodine 1.4 g Calcium iodate 

Cobalt 0.4 g Cobalt carbonate 

a Per ton (2,000 lb) of finished product 
D&C Yellow No. 11, NTP TR 463 181 

TABLE I3 

Nutrient Composition of NIH-07 Rat and Mouse Ration 

Mean ± Standard 

Nutrient Deviation Range Number of Samples 

Protein (% by weight) 23.68 ± 0.54 22.5 – 25.2 27 

Crude fat (% by weight) 5.25 ± 0.23 4.80 – 5.80 27 

Crude fiber (% by weight) 3.56 ± 0.43 2.60 – 4.30 27 

Ash (% by weight) 6.48 ± 0.19 6.12 – 6.97 27 

Amino Acids (% of total diet) 

Arginine 1.280 ± 0.083 1.110 – 1.390 11 

Cystine 0.308 ± 0.071 0.181 – 0.400 11 

Glycine 1.158 ± 0.048 1.060 – 1.220 11 

Histidine 0.584 ± 0.027 0.531 – 0.630 11 

Isoleucine 0.917 ± 0.033 0.867 – 0.965 11 

Leucine 1.975 ± 0.051 1.850 – 2.040 11 

Lysine 1.274 ± 0.049 1.200 – 1.370 11 

Methionine 0.437 ± 0.109 0.306 – 0.699 11 

Phenylalanine 0.999 ± 0.120 0.665 – 1.110 11 

Threonine 0.904 ± 0.058 0.824 – 0.985 11 

Tryptophan 0.218 ± 0.153 0.107 – 0.671 11 

Tyrosine 0.685 ± 0.094 0.564 – 0.794 11 

Valine 1.086 ± 0.055 0.962 – 1.170 11 

Essential Fatty Acids (% of total diet) 

Linoleic 2.407 ± 0.227 1.830 – 2.570 10 

Linolenic 0.259 ± 0.065 0.100 – 0.320 10 

Vitamins 

Vitamin A (IU/kg) 6,821 ± 1,531 4,290 – 12,540 27 

Vitamin D (IU/kg) 4,450 ± 1,382 3,000 – 6,300 4 

-Tocopherol (ppm) 36.12 ± 9.15 22.50 – 48.9 10 

Thiamine (ppm) 18.81 ± 2.11 15.0 – 25.0 27 

Riboflavin (ppm) 7.83 ± 0.923 6.10 – 9.00 11 

Niacin (ppm) 98.64 ± 25.5 65.0 – 150.0 10 

Pantothenic acid (ppm) 30.55 ± 3.52 23.0 – 34.6 11 

Pyridoxine (ppm) 9.11 ± 2.53 5.60 – 14.0 11 

Folic acid (ppm) 2.46 ± 0.63 1.80 – 3.70 11 

Biotin (ppm) 0.268 ± 0.047 0.190 – 0.354 11 

Vitamin B12 (ppb) 40.5 ± 19.1 10.6 – 65.0 11 

Choline (ppm) 2,991 ± 382 2,300 – 3,430 10 

Minerals 

Calcium (%) 1.18 ± 0.09 1.02 – 1.37 27 

Phosphorus (%) 0.94 ± 0.046 0.800 – 1.03 27 

Potassium (%) 0.886 ± 0.063 0.772 – 0.971 9 

Chloride (%) 0.529 ± 0.087 0.380 – 0.635 9 

Sodium (%) 0.316 ± 0.033 0.258 – 0.371 11 

Magnesium (%) 0.166 ± 0.010 0.148 – 0.181 11 

Sulfur (%) 0.272 ± 0.059 0.208 – 0.420 10 

Iron (ppm) 350.5 ± 87.3 255.0 – 523.0 11 

Manganese (ppm) 92.48 ± 5.14 81.7 – 99.4 11 

Zinc (ppm) 59.33 ± 10.2 46.1 – 81.6 11 

Copper (ppm) 11.81 ± 2.50 8.09 – 15.4 11 

Iodine (ppm) 3.54 ± 1.19 1.52 – 5.83 10 

Chromium (ppm) 1.66 ± 0.46 0.85 – 2.09 11 

Cobalt (ppm) 0.76 ± 0.23 0.49 – 1.15 7 
182 D&C Yellow No. 11, NTP TR 463
 

TABLE I4 

Contaminant Levels in NIH-07 Rat and Mouse Ration a 

Mean ± Standard 

Deviationb Range Number of Samples 

Contaminants
 

Arsenic (ppm) 0.40 ± 0.18 0.10 – 0.80 27
 

Cadmium (ppm) 0.10 ± 0.07 0.05 – 0.20 27
 

Lead (ppm) 0.27 ± 0.21 0.10 – 1.10 27
 

Mercury (ppm) 0.02 ± 0.01 0.02 – 0.50 27
 c Selenium (ppm) 0.33 ± 0.10 0.10 – 0.44 26
 d Aflatoxins (ppb) <5.0 26
 e Nitrate nitrogen (ppm) 10.77 ± 4.92 1.80 – 20.0 27
 e Nitrite nitrogen (ppm) 0.22 ± 0.16 0.10 – 0.60 27
 f BHA (ppm) 1.42 ± 0.90 1.00 – 4.00 26
 f BHT (ppm) 1.31 ± 1.19 1.00 – 7.00 26
 

Aerobic plate count (CFU/g) 109,767 ± 105,017 4,700 – 380,000 27
 

Coliform (MPN/g) 17.7 ± 20.5 3.00 – 93.00 27
 

Escherichia coli (MPN/g) 3.3 ± 1.2 3.0 – 9.0 27
 

Salmonella (MPN/g) Negative 27
 

Total nitrosoamines (ppb)g 7.00 ± 2.10 3.90 – 13.70 27
 

N-Nitrosodimethylamine (ppb)g 5.28 ± 1.45 2.90 – 9.40 27
 

N-Nitrosopyrrolidine (ppb)g 1.72 ± 1.01 1.00 – 4.70 27
 

Pesticides (ppm) 

-BHC <0.01 27
 

-BHC <0.02 27
 

-BHC <0.01 27
 

-BHC <0.01 27
 

Heptachlor <0.01 27
 

Aldrin <0.01 27
 

Heptachlor epoxide <0.01 27
 

DDE <0.01 27
 

DDD <0.01 27
 

DDT <0.01 27
 

HCB <0.01 27
 

Mirex <0.01 27
 

Methoxychlor <0.05 27
 

Dieldrin <0.01 27
 

Endrin <0.01 27
 

Telodrin <0.01 27
 

Chlordane <0.05 27
 

Toxaphene <0.10 27
 

Estimated PCBs <0.20 27
 

Ronnel <0.01 27
 

Ethion <0.02 27
 

Trithion <0.05 27
 

Diazinon <0.10 27
 

Methyl parathion <0.02 27
 

Ethyl parathion <0.02 27
 

Malathion 0.27 ± 0.21 0.05 – 0.84 27
 

Endosulfan I <0.01 27
 

Endosulfan II <0.01 27
 

Endosulfan sulfate <0.03 27
 

a CFU=colony forming units. MPN=most probable number. BHC=hexachlorocyclohexane or benzene hexachloride. b For values less than the limit of detection, the detection limit is given as the mean.
 

No selenium measurement was recorded for the lot milled 5 May 1990. d No aflatoxin measurement was recorded for the lot milled 2 October 1989. e Sources of contamination: alfalfa, grains, and fish meal. f Sources of contamination: soy oil and fish meal. No BHA or BHT measurements were recorded for the lot milled 1 November 1989. 

g All values were corrected for percent recovery. 

c 
183 

APPENDIX J
 

SENTINEL ANIMAL PROGRAM
 

METHODS .................................................................................... 184
 

RESULTS ..................................................................................... 185
 
184 D&C Yellow No. 11, NTP TR 463 

SENTINEL ANIMAL PROGRAM 

METHODS 

Rodents used in the Carcinogenesis Program of the National Toxicology Program are produced in optimally 

clean facilities to eliminate potential pathogens that may affect study results. The Sentinel Animal Program is 

part of the periodic monitoring of animal health that occurs during the toxicologic evaluation of chemical 

compounds. Under this program, the disease state of the rodents is monitored via serology on sera from extra 

(sentinel) animals in the study rooms. These animals and the study animals are subject to identical 

environmental conditions. The sentinel animals come from the same production source and weanling groups as 

the animals used for the studies of chemical compounds. 

Serum samples were collected from randomly selected rats during the reproductive toxicity and 2-year studies. 

Blood from each animal was collected and allowed to clot, and the serum was separated. The samples were 

processed appropriately and sent to Microbiological Associates, Inc. (Bethesda, MD), for determination of 

antibody titers. The laboratory serology methods and viral agents for which testing was performed are tabulated 

below; the times at which blood was collected during the studies are also listed. 

Method and Test Time of Analysis 

Reproductive Toxicity Study 

ELISA 

PVM (pneumonia virus of mice) Study termination 

RCV/SDA (rat coronavirus/sialodacryoadenitis virus) Study termination 

Sendai Study termination 

Immunofluorescence Assay 

RCV/SDA Study termination 

Hemagglutination Inhibition 

H-1 (Toolan’s H-1 virus) Study termination 

KRV (Kilham rat virus) Study termination 

2-Year Study 

ELISA 

Mycoplasma arthritidis Study termination 

Mycoplasma pulmonis Study termination 

PVM 6, 12, and 18 months, study termination 

RCV/SDA 6, 12, and 18 months, study termination 

Sendai 6, 12, and 18 months, study termination 

Immunofluorescence Assay 

RCV/SDA 12 Months 

Hemagglutination Inhibition 

H-1 6, 12, and 18 months, study termination 

KRV 6, 12, and 18 months, study termination 
D&C Yellow No. 11, NTP TR 463 185 

RESULTS 

For the reproductive toxicity study in rats, all serology test results were negative. Two female rats had positive titers 

to M. arthritidis at the end of the 2-year study. 

Further evaluation of samples positive for M. arthritidis by immunoblot and Western blot procedures indicated that 

the positive titers may have been due to a cross reaction with antibodies of nonpathogenic Mycoplasma or other 

agents. Only sporadic samples were positive, and there were no clinical findings or histopathologic changes of M. 

arthritidis infection in rats with positive titers. Accordingly, sporadic M. arthritidis-positive titers were considered to 

be false positives. 
186 D&C Yellow No. 11, NTP TR 463