NTP TECHNICAL REPORT
 

ON THE
 

TOXICOLOGY AND CARCINOGENESIS
 

STUDIES OF
 

TECHNICAL GRADE
 

SODIUM XYLENESULFONATE
 

(CAS NO. 1300-72-7) 

IN F344/N RATS AND B6C3F1 MICE

(DERMAL STUDIES) 

NATIONAL TOXICOLOGY PROGRAM
 

P.O. Box 12233
#

Research Triangle Park, NC 27709
#

June 1998 

NTP TR 464
#

NIH Publication No. 98-3380
#

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. 

Listings of all published NTP reports and ongoing studies are available from NTP Central Data

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

The Abstracts and other study information for 2-year studies are 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
 

TECHNICAL GRADE
 

SODIUM XYLENESULFONATE
 

(CAS NO. 1300-72-7) 

IN F344/N RATS AND B6C3F1 MICE

(DERMAL STUDIES) 

NATIONAL TOXICOLOGY PROGRAM
 

P.O. Box 12233
#

Research Triangle Park, NC 27709
#

June 1998 

NTP TR 464
#

NIH Publication No. 98-3380
#

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES
 

Public Health Service
 

National Institutes of Health
 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

2 Sodium Xylenesulfonate, NTP TR 464 

CONTRIBUTORS
 

National Toxicology Program

Evaluated and interpreted results and reported findings

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

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

J.K. Haseman, 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 for 17-day studies in

rats and mice

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

T. Makovec, D.V.M. 

Southern Research Institute

Conducted studies, evaluated pathology findings for 14-week studies

in rats and mice

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

D.R. Farnell, D.V.M., Ph.D. 

R.B. Thompson, D.V.M., Ph.D. 

Battelle Columbus Laboratories

Conducted studies, evaluated pathology findings for 2-year studies in

rats and mice

P.J. Kurtz, Ph.D., Principal Investigator

M.R. Hejtmancik, Ph.D. 

A.W. Singer, D.V.M. 

J.D. Toft, II, D.V.M., M.S. 

Experimental Pathology Laboratories, Inc. Provided pathology quality assurance

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

E.T. Gaillard, D.V.M., M.S. 

W.F. Mackenzie, D.V.M., M.S. 

Dynamac Corporation

Prepared quality assurance audits

S. Brecher, Ph.D., Principal Investigator

NTP Pathology Working Group

Evaluated slides, prepared pathology report for 14-week studies in

rats and mice (13 June 1989) 

S. Grumbein, D.V.M., Ph.D., Chairperson

Pathology Associates, Inc. 

M.R. Elwell, D.V.M., Ph.D. National Toxicology Program

J.R. Leininger, D.V.M., Ph.D. National Toxicology Program

W.F. Mackenzie, D.V.M., M.S. Experimental Pathology Laboratories, Inc. 

A.W. Macklin, D.V.M., Ph.D. Burroughs Wellcome Co. 

B. Palate, D.V.M., Observer

Searle European Development Centre

Evaluated slides, prepared pathology report for 2-year studies in rats

and mice (1 June 1995) 

D.G. Goodman, V.M.D., Chairperson

PATHCO, Inc. 

P. Carthew, Ph.D., Observer

National Toxicology Program

A. Enomoto, D.V.M. National Toxicology Program

J. Everitt, D.V.M. 

Chemical Industry Institute of Toxicology

E.T. Gaillard, D.V.M., M.S. Experimental Pathology Laboratories, Inc. 

J.R. Hailey, D.V.M. National Toxicology Program

R.A. Herbert, D.V.M., Ph.D. National Toxicology Program

A. Radovsky, D.V.M., Ph.D. National Toxicology Program

M. Torii, D.V.M., Ph.D., Observer

National Toxicology Program

Analytical Sciences, Inc. Provided statistical analyses

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

S.R. Lloyd, M.S. 

N.G. Mintz, B.S. 

Biotechnical Services, Inc. Prepared Technical Report

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

L.M. Harper, B.S. 

E.S. Rathman, M.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 ............................................... 15 
 #

RESULTS .............................................................. 25 
 #

DISCUSSION AND CONCLUSIONS ........................................... 47 
 #

REFERENCES .......................................................... 51 
 #

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

of Sodium Xylenesulfonate ....................................... 55 
 #

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

of Sodium Xylenesulfonate ....................................... 97 
 #

APPENDIX C Summary of Lesions in Male Mice in the 2-Year Dermal Study
 

of Sodium Xylenesulfonate ....................................... 127 
 #

APPENDIX D Summary of Lesions in Female Mice in the 2-Year Dermal Study
 

of Sodium Xylenesulfonate ....................................... 161 
 #

APPENDIX E Genetic Toxicology ............................................. 201 
 #

APPENDIX F Organ Weights and Organ-Weight-to-Body-Weight Ratios .................. 215 
 #

APPENDIX G Hematology and Clinical Chemistry Results ............................ 221 
 #

APPENDIX H Chemical Characterization and Dose Formulation Studies .................. 227 
 #

APPENDIX I Doses, Body Weights, and Dose Concentrations ......................... 239 
 #

APPENDIX J Ingredients, Nutrient Composition, and Contaminant Levels
 

in NIH-07 Rat and Mouse Ration .................................. 243 
 #

APPENDIX K Sentinel Animal Program ........................................ 247 
 #

APPENDIX L 1 Impact of Helicobacter hepaticus Infection in B6C3F Mice

from 12 NTP 2-Year Carcinogenesis Studies ........................... 251 
 #
4 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5 

ABSTRACT
 

SODIUM XYLENESULFONATE

CAS No. 1300-72-7
#

Chemical Formula: (CH ) C H SO ! Na 32 6 3 3 + Molecular Weight: 208.2

Synonyms: Benzenesulfonic acid, dimethyl-, sodium salt; sodium dimethylbenzenesulfonate; xylenesulfonic acid, sodium salt
 

Trade names: Conco SXS; Cyclophil; SXS 30; Eletesol SX 30; Naxonate; Naxonate G; Richonate SXS; Stepanate SXS; Stepanate X; SXS 40;
#

Ultrawet 40SX

Sodium xylenesulfonate is used as a hydrotrope, an

organic compound that increases the ability of water

to dissolve other molecules. Sodium xylenesulfonate

is a component in a variety of widely used shampoos

and liquid household detergents where it can

constitute up to 10% of the total solution. Because of

its widespread use, the potential for human exposure

to sodium xylenesulfonate is great. Male and female

F344/N rats and B6C3F1 mice were administered

sodium xylenesulfonate in water or 50% ethanol

dermally for 17 days, 14 weeks, or 2 years. Genetic

toxicology studies were conducted in Salmonella

typhimurium, L5178Y mouse lymphoma cells, and

cultured Chinese hamster ovary cells. 

17-DAY STUDY IN RATS

Groups of five male and five female rats were

administered 300 μL of 0, 5, 15, 44, 133, or

400 mg/mL sodium xylenesulfonate in distilled water

by dermal application 5 days per week for 17 days. 

All rats survived to the end of the study. Final mean

body weights and body weight gains of dosed rats

were similar to those of the control groups. Dermal

applications of 300 μL of 5, 15, 44, 133, and

400 mg/mL delivered average daily doses of

approximately 10, 30, 90, 260, and 800 mg sodium

xylenesulfonate/kg body weight to males and 13, 40, 

120, 330, and 1,030 mg/kg to females. Clinical

findings generally involved the skin of dosed animals

and included tan or brown skin discoloration and

crusty white deposits (presumed to be dried chemical) 

at the site of application. Neither of these

observations were considered significant findings. 

The relative liver weights of 133 and 400 mg/mL

male and female rats were significantly greater than

those of the control groups, but the absolute liver

weights were not increased and the biological

significance of the relative differences in liver weight

was unclear. In males and females, the few lesions

observed grossly and microscopically were generally

attributed to repeated clipping and were not

considered related to chemical administration. 

17-DAY STUDY IN MICE

Groups of five male and five female mice were

administered 100 μL of 0, 5, 15, 44, 133, or
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

6 Sodium Xylenesulfonate, NTP TR 464 

400 mg/mL sodium xylenesulfonate in distilled water

by dermal application 5 days per week for 17 days. 

All mice survived to the end of the study. Final mean

body weights and body weight gains of dosed mice

were similar to those of the controls. Dermal

applications of 5, 15, 44, 133, and 400 mg/mL

delivered average daily doses of approximately, 

20, 60, 190, 540, and 1,600 mg sodium xylenesulfonate/kg body weight to males and 26, 80, 220, 

680, and 2,000 mg/kg to females. Clinical findings

included crusty white deposits (presumed to be dried

chemical) at the site of application in two 133 mg/mL

males and in all 400 mg/mL males and females. The

absolute and relative liver weights of 15 and

44 mg/mL males and 400 mg/mL males and females

were significantly greater than those of the control

groups, but the biological significance of these

differences was unclear. The few skin lesions

observed grossly and microscopically in males and

females were generally attributed to repeated clipping

and were not considered related to chemical

administration. 

14-WEEK STUDY IN RATS

Groups of 10 male and 10 female rats were

administered 300 μL of 0, 5, 15, 44, 133, or

400 mg/mL sodium xylenesulfonate in 50% ethanol

by dermal application for 14 weeks. For special

hematology and clinical pathology studies, additional

groups of 10 male and 10 female rats were

administered 0, 5, 15, 44, 133, or 400 mg/mL sodium

xylenesulfonate in 50% ethanol by dermal application

for 14 weeks. All rats survived to the end of the

study. Final mean body weights and body weight

gains of dosed male and female rats were similar to

those of the control groups. Dermal applications of 5, 

15, 44, 133, and 400 mg/mL delivered average daily

doses of approximately 6, 20, 60, 170, and 500 mg

sodium xylenesulfonate/kg body weight to males and

10, 30, 90, 260, and 800 mg/kg to females. The only

notable clinical finding was brown discoloration of the

skin at the site of application in dosed animals. 

Hematology and clinical chemistry parameters of

dosed groups of males and females were significantly

different from those of the controls in several

instances, but these differences were sporadic and did

not demonstrate a treatment relationship. The

absolute and relative liver weights of males receiving

44, 133, or 400 mg/mL were significantly less than

those of the control group, but the biological

significance of these differences was unclear, and

there were no treatment-related histopathologic effects

in the liver. There were no significant differences in

liver weights in female rats. 

Minimal hyperplasia of the epidermis at the site of

application occurred in both male and female rats in

the control group as well as most dosed groups. The

incidence of epidermal hyperplasia in 400 mg/mL

males was possibly chemical related. 

14-WEEK STUDY IN MICE

Groups of 10 male and 10 female mice were

administered 100 μL of 0, 5, 15, 44, 133, or

400 mg/mL sodium xylenesulfonate in 50% ethanol

by dermal application for 14 weeks. There were no

chemical-related deaths. The mean body weight gain

of the 400 mg/mL males was significantly greater than

that of the control group. Dermal applications of 5, 

15, 44, 133, and 400 mg/mL delivered average daily

doses of approximately 17, 40, 140, 440, and

1,300 mg sodium xylenesulfonate/kg body weight to

males and 20, 60, 170, 530, and 1,630 mg/kg to

females. There were no clinical findings related to

sodium xylenesulfonate administration. 

Epidermal hyperplasia occurred in one 44 mg/mL

female, two 133 mg/mL males, five 400 mg/mL

males, and four 400 mg/kg females. Hyperplasia of

the epidermis in 400 mg/mL males and females was

probably related to chemical administration. 

Chronic inflammation of the skin occurred primarily

in the control groups of males and females. These

lesions consisted of mononuclear inflammatory cells

in the dermis. 

2-YEAR STUDY IN RATS

Groups of 50 male and 50 female rats were dermally

administered 0, 60, 120, or 240 mg sodium xylenesulfonate/kg body weight in 50% ethanol for

104 weeks. 

Survival, Body Weights, and Clinical Findings

Survival of dosed males and females was similar to

that of the control groups. Mean body weights of

dosed males and females were similar to those of the
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

__________

 

 

Sodium Xylenesulfonate, NTP TR 464 7 

controls throughout the study. In male groups, there

were no clinical findings considered treatment related. 

In females, clinical findings were limited to irritation

at the site of application in one control female, four

120 mg/kg females, and two 240 mg/kg females. 

Pathology Findings

There were no neoplasms at any site (including the

skin) that were considered treatment related. 

Low incidences of hyperplasia of the epidermis at the

site of application occurred in males in the 60, 120, 

and 240 mg/kg groups. Low incidences of

hyperplasia of the epidermis at the site of application

also occurred in females in the 120 and 240 mg/kg

groups, and they occurred with a significant positive

trend. Low incidences of hyperplasia of the

sebaceous gland occurred in control and 60 mg/kg

males and in control, 120 mg/kg, and 240 mg/kg

females. 

2-YEAR STUDY IN MICE

Groups of 50 male and 50 female mice were dermally

administered 0, 182, 364, or 727 mg sodium xylenesulfonate/kg body weight in 50% ethanol for 104 to

105 weeks. 

Survival, Body Weights, and Clinical Findings

Survival of dosed males and females was similar to

that of the control groups. Mean body weights of

dosed males and females were generally similar to

those of the controls throughout the study; however, 

the mean body weights of 727 mg/kg females were

greater than those of the control group from week 85 

to week 97. With the exception of irritation at the site

of application in one 364 mg/kg female, there were no

clinical findings related to sodium xylenesulfonate

administration. 

Pathology Findings

There were no neoplasms at any site (including the

skin) that were considered treatment related. 

Hyperplasia of the epidermis occurred in control, 

364 mg/kg, and 727 mg/kg males and in control and

dosed females. In male mice, the incidences occurred

with a significant positive trend. Focal ulceration

occurred in one 727 mg/kg male and in one female in

each dose group. In males and females from control

and dosed groups, the incidences of hepatocellular

adenoma, hepatocellular carcinoma, and hepatocellular adenoma or carcinoma (combined) were

generally higher than those expected by spontaneous

occurrence. The incidences of hepatocellular

neoplasms in some groups of males and females

exceeded the NTP historical control range. Male

mice had a pattern of nonneoplastic liver lesions along

with silver stained positive helical organisms within

the liver which suggests an infection with

Helicobacter hepaticus. The findings in this study of

sodium xylenesulfonate were not considered to have

been significantly impacted by the infection with

H. hepaticus or its associated hepatitis. 

GENETIC TOXICOLOGY

Sodium xylenesulfonate was not mutagenic in

Salmonella typhimurium strain TA98, TA100, 

TA1535, or TA1537 with or without induced liver S9. 

Equivocal results were obtained in a mutation assay

with mouse lymphoma cells in the presence of induced

S9; no evidence of mutagenicity was noted without S9 

in this assay. In cytogenetic tests with sodium xylenesulfonate in cultured Chinese hamster ovary cells, 

significant increases in sister chromatid exchanges

were observed in the absence of S9 only, and no

increases in chromosomal aberrations were observed

with or without S9. 

CONCLUSIONS

Under the conditions of these 2-year dermal studies, 

there was no evidence of carcinogenic activity* of

sodium xylenesulfonate in male or female F344/N rats

administered 60, 120, or 240 mg/kg or in male or

female B6C3F1 mice administered 182, 364, or

727 mg/kg. 

Increased incidences of epidermal hyperplasia in

female rats and male mice may have been related to

exposure to sodium xylenesulfonate. 

* 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 Sodium Xylenesulfonate, NTP TR 464 

Summary of the 2-Year Carcinogenesis and Genetic Toxicology Studies of Sodium Xylenesulfonate

Male Female Male Female
 

F344/N Rats F344/N Rats B6C3F Mice B6C3F Mice
 1 1 

Doses	 0, 60, 120, or 0, 60, 120, or 0, 182, 364, or 0, 182, 364, or

240 mg/kg in 50% 240 mg/kg in 50% 727 mg/kg in 50% 727 mg/kg in 50% 

ethanol applied ethanol applied ethanol applied ethanol applied

dermally dermally dermally dermally

Body weights	 Dosed groups similar Dosed groups similar Dosed groups similar Dosed groups similar

to control group to control group to control group to control group

2-Year survival	 7/50, 17/50, 9/50, 22/50, 16/50, 17/50, 32/50, 37/50, 39/50, 31/50, 32/49, 32/50, 

10/50 16/50 35/50 36/50 rates

Nonneoplastic None None None None

effects

Neoplastic effects	 None None None None

Uncertain findings None	 Skin (site of Skin (site of None

application): application): 

epidermal hyperplasia epidermal hyperplasia

(1/50, 0/50, 4/50, (1/50, 0/50, 4/50, 

5/50) 5/50) 

Level of evidence No evidence No evidence No evidence No evidence

of carcinogenic

activity

Genetic toxicology

Salmonella typhimurium gene mutations: Negative with and without S9 in strains TA98, TA100, TA1535, and TA1537 

Mouse lymphoma mutagenicity Equivocal with S9; negative without S9 

Sister chromatid exchanges

Cultured Chinese hamster ovary cells in vitro: Negative with S9; positive without S9
#

Chromosomal aberrations
 

Cultured Chinese hamster ovary cells in vitro: Negative with and without S9
#
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 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 evidence of 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 study of 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 long-term 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 Sodium Xylenesulfonate, NTP TR 464 

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 technical grade sodium

xylenesulfonate 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 the 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. 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., Principal Reviewer 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., Principal Reviewer

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.* National Cancer Institute

Department of Pathology Frederick, MD

Northwestern University Medical School

Chicago, IL

* Did not attend
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 11 

SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS
 

On 5 December 1995, the draft Technical Report on state any potential dose or absorption effects that

the toxicology and carcinogenesis studies of sodium occurred from changing volumes as well as vehicles

xylenesulfonate received public review by the from the 17-day studies to the 14-week and 2-year

National Toxicology Program’s Board of Scientific studies, and comment on the relevance of these studies

Counselors’ Technical Reports Review Subcommittee. to human exposures. Dr. Radovsky said sodium

The review meeting was held at the National Institute xylenesulfonate was more soluble in water than in

of Environmental Health Sciences, Research Triangle ethanol, but ethanol may have enhanced skin

Park, NC. penetration more than water. She said that relevance

to human exposure would be speculative on her part. 

Dr. A. Radovsky, NIEHS, introduced the toxicology

and carcinogenesis studies of sodium xylenesulfonate Dr. Tyson, the third principal reviewer, agreed with

by discussing the uses of the chemical, describing the the proposed conclusions. He also questioned the use

experimental design, reporting on survival and body of ethanol as the vehicle noting the association of

weight effects, and commenting on possible chemical- dermally applied ethanol with induction of monorelated nonneoplastic lesions in female rats and male nuclear cell leukemia in F344/N rats. 

mice. Dr. Radovsky reported that the greater than

normal incidence of hepatocellular neoplasms in Dr. W.T. Allaben, NCTR, asked for comment on the

control and treated male mice could be attributed to poor survival in male rats. Dr. J.R. Bucher, NIEHS, 

infection with Helicobacter bacteria. Increased said that male rat survival has declined primarily

incidences of hepatocellular neoplasms in female mice because of increases in nephropathy, body weight, 

could not be associated with Helicobacter with and incidence of pituitary adenoma. Dr. G.N. Rao, 

certainty. The proposed conclusions were no NIEHS, commented that survival of rats is lower

evidence of carcinogenic activity of sodium xylene- when they are individually housed as opposed to

sulfonate in male and female F344/N rats and B6C3F group-housed. Dr. J.K. Haseman, NIEHS, said that 1 

mice. survival in this study was similar to that in other

dermal studies using individual housing. Dr. Ryan

Dr. Carlson, a principal reviewer, agreed with the asked whether there could be a correlation between

proposed conclusions. He commented that he would mice with Helicobacter and those with neoplastic

not have used ethanol as vehicle for application of the lesions. Dr. J.R. Hailey, NIEHS, said that although

chemical. not all of the animals had been examined, for those

that had, there was a good correlation. Dr. Haseman

Dr. Goldsworthy, the second principal reviewer, commented that in the previous study with

agreed in principle with the proposed conclusions Helicobacter, animals with liver neoplasms had the

provided there were further clarification and more severe nonneoplastic lesions that were indicative

documentation on the role of Helicobacter in the of the infection. Dr. Goldsworthy thought the

mouse liver neoplasm responses. He said that the infection could be a confounding factor in the

report needed to better address the response in interpretation of the liver neoplasms in mice. Dr. 

females and the effects observed in males and females Haseman said that while total liver neoplasm rates in

in a comprehensive manner to ensure that the this study were above expected rates, they were

responses are properly interpreted as nontreatment generally similar across groups, yielding no evidence

related. Dr. Radovsky responded that several other of a chemical-related increase. 

studies with Helicobacter infection were completed

and would be reviewed at the next review meeting. Dr. F. Mirer, Health and Safety Department, United

Hopefully, firmer conclusions could then be drawn Auto Workers Union, had submitted a statement

about the association of liver neoplasm response and which, at his request, Dr. L.G. Hart, NIEHS, read

infection in B6C3F mice (Appendix L). into the record. Dr. Mirer opined that the studies 1 

Dr. Goldsworthy said the report should more clearly were inadequate to address the carcinogenicity of
 

 

 

 

 

 

 

 

 

 

 

 

12 Sodium Xylenesulfonate, NTP TR 464 

sodium xylenesulfonate in humans. He based his discussed and with the conclusions as written for male

assessment on: (1) not high enough a dose to and female rats and mice, no evidence of carcinogenic

approach a maximum tolerated dose (MTD); (2) likely activity. Dr. Tyson seconded the motion. 

poor absorption of such an ionic material through Dr. Goldsworthy offered an amendment that a

intact skin; and (3) wrong route of exposure to statement be added to the Abstract that mice were

estimate human risk, i.e., inhalation exposure should infected with Helicobacter. Dr. Carlson agreed to the

have been used. amendment, and the amended motion was accepted by

six yes votes to one no vote (Dr. Russo). Dr. Allaben

asked that a short paragraph be added to the

Dr. Carlson moved that the Technical Report on discussion regarding individual animal housing and

sodium xylenesulfonate be accepted with the revisions poor survival. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

13 

INTRODUCTION
 

SODIUM XYLENESULFONATE

CAS No. 1300-72-7 

Chemical Formula: (CH ) C H SO ! Na 32 6 3 3 

+ Molecular Weight: 208.2

Synonyms: Benzenesulfonic acid, dimethyl-, sodium salt; sodium dimethylbenzenesulfonate; xylenesulfonic acid, sodium salt

Trade names: Conco SXS; Cyclophil; SXS 30; Eletesol SX 30; Naxonate; Naxonate G; Richonate SXS; Stepanate SXS; Stepanate X; 

SXS 40; Ultrawet 40SX

CHEMICAL AND PHYSICAL PROPERTIES

Technical grade sodium xylenesulfonate contains

approximately 35% sodium ethylbenzenesulfonate and

11.5%, 38%, and 15.5% of the ortho-, meta-, and

para- isomers, respectively, of sodium xylenesulfonate (Mausner and Sosis, 1962). In water at

20E C, the para- isomer of sodium xylenesulfonate

has a solubility of 170 mg/mL, the ortho- isomer has

a solubility of 190 mg/mL, and the meta- isomer has

a solubility of more than 400 mg/mL. Solubility of

sodium xylenesulfonate in ethanol is less than that in

water. Sodium xylenesulfonate is stable in water

under normal laboratory conditions and is incompatible with strong acids and oxidizers (RTECS, 

1991). 

PRODUCTION, USE, 

AND HUMAN EXPOSURE

In 1992, over 27.3 × 10 6 kg of sodium xylenesulfonate was produced in the United States by

10 manufacturers (USITC, 1994). The major use of

sodium xylenesulfonate is as a hydrotrope, an organic

compound that increases and maintains the solubility

of other organic compounds in aqueous solutions. It

is used in liquid household detergents and shampoos, 

in degreasing compounds and printing pastes used in

the textile industry, in agents used to extract pentosans

and lignin in the paper industry, and as a glue additive

in the leather industry. 

According to the National Occupational Exposure

Survey, approximately 788,092 workers in the United

States were potentially exposed to sodium xylenesulfonate during the years 1981 to 1983 (NIOSH, 

1990). In shampoos, sodium xylenesulfonate can

constitute up to 0.2% of the total solution (Hunting, 

1983). Commonly used household products such as

disinfectants, floor care products, and kitchen cleaners

and degreasers contain sodium xylenesulfonate in

concentrations up to 10% (Phil Tham, Reckitt and

Colman, Inc., personal communication, 1995). 

Because many of these shampoos and household

products are often used daily, there is widespread

potential for human exposure. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

14 Sodium Xylenesulfonate, NTP TR 464 

ABSORPTION, DISTRIBUTION, 

METABOLISM, AND EXCRETION

No information on the absorption, distribution, 

metabolism, and excretion of sodium xylenesulfonate

in experimental animals or in humans was found in

the literature. Although no information is available

about absorption of sodium xylenesulfonate specifically, skin penetration has been shown to be low in

compounds that have a polar group, such as the

sulfonate group in sodium xylenesulfonate, which can

interact with polar groups in the stratum corneum

(Grandjean et al., 1988). In general, when absorbed, 

sulfonates are quickly distributed throughout the body, 

but are also readily excreted (Patty’s, 1981). 

TOXICITY

No information specific to the toxicity of sodium

xylenesulfonate in experimental animals or in humans

was found in the literature. In general, the sodium

salts of arylsulfonic acids are considered to be of

relatively low toxicity (Patty’s, 1981). The reported

minimum lethal gavage dose for sodium toluenesulfonate, a closely related compound, is 12,000 mg/kg

in albino rats and 10,000 mg/kg in albino mice

(Kondratiuk, 1983). 

CARCINOGENICITY

No information specific to the carcinogenicity of

sodium xylenesulfonate in experimental animals or in

humans was found in the literature. 

GENETIC TOXICITY

Sodium xylenesulfonate was not mutagenic in

Salmonella typhimurium strains TA98, TA100, 

TA1535, and TA1537 when tested at concentrations

up to 10,000 μg/plate with and without Aroclor 1254-

induced male Sprague-Dawley rat or Syrian hamster

liver S9 (Zeiger et al., 1987). 

STUDY RATIONALE

Sodium xylenesulfonate was nominated by the

National Cancer Institute from a soap and detergent

class study because of its high annual production and

its presence in widely used consumer products. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

15 

MATERIALS AND METHODS
 

PROCUREMENT

AND CHARACTERIZATION

OF SODIUM XYLENESULFONATE

Sodium xylenesulfonate was obtained from Ruetgers

Nease Chemical Company (State College, PA) in one

lot (R092085), which was used for the 17-day, 

14-week, and 2-year studies. Identity and purity

analyses were conducted by the analytical chemistry

laboratory, Midwest Research Institute (Kansas City, 

MO) (Appendix H). Reports on analyses performed

in support of the sodium xylenesulfonate studies are

on file at the National Institute of Environmental

Health Sciences (NIEHS). 

The chemical, a white powder, was identified as

sodium xylenesulfonate by infrared, ultraviolet/ 

visible, and nuclear magnetic resonance spectroscopy. 

The purity was determined by elemental analysis, Karl

Fischer water analysis, functional group titration, 

thin-layer chromatography, and high-performance

liquid chromatography. 

Elemental analyses for carbon and hydrogen were in

agreement with the theoretical values for sodium

xylenesulfonate. Calculated on the basis of 5.15% 

water and 3.82% sodium sulfate, results for sulfur

were slightly higher than theoretical values and results

for sodium were slightly lower than theoretical values. 

Karl Fischer water analysis indicated 5.15% ± 0.07% 

water. Functional group titration for ionic sulfate

indicated 3.82% ± 0.04% sodium sulfate. Thin-layer

chromatography by one system indicated a major spot

and a slight trace impurity; a second system indicated

a major spot only. High-performance liquid chromatography by one system indicated a major peak and

five impurities with areas totaling 39.2% relative to

the major peak; a second system indicated an additional impurity with a peak area of 9.6% relative to

the major peak. Under these chromatographic conditions, sodium 2,4- and 2,5-xylenesulfonate coeluted

with the major peak. Analysis of nuclear magnetic

spectra suggests that one of the impurities may have

been ethyl benzenesulfonate. 

To ensure stability of the bulk chemical during the

17-day and 14-week studies, sodium xylenesulfonate

was stored in glass bottles with Teflon®-lined caps or

double bagged in metal drums at room temperature in

the dark. During the 2-year studies, the bulk chemical was stored in amber glass bottles at room temperature in the dark. 

PREPARATION AND ANALYSIS

OF DOSE FORMULATIONS

The dose formulations for the 17-day studies were

prepared twice during the study by stirring the appropriate quantities of sodium xylenesulfonate into

deionized water which was then brought to the desired

concentration by the further addition of deionized

water (Table H1). The dose formulations for the

14-week studies were prepared every 2 weeks by

mixing the appropriate amount of sodium xylenesulfonate with 50% ethanol (in deionized water) which

was then brought to the desired concentration by the

further addition of 50% ethanol (Table H1). The 5 

and 15 mg/mL doses were stirred to solution; the 44, 

133, and 400 mg/mL doses were stirred to suspension. Dose formulations in the 2-year studies of

sodium xylenesulfonate were prepared every 2 to

3 weeks by mixing the appropriate weight of sodium

xylenesulfonate in deionized water (Table H1). A

suspension was formed by shaking, and then the

mixture was brought to the desired concentrations of

ethanol and sodium xylenesulfonate by the addition of

95% ethanol. The resulting suspension was the

required dose in 50% ethanol. Dose formulations in

the 17-day, 14-week, and 2-year studies were stored

at room temperature in glass bottles with Teflon®-

lined lids in the dark for up to 3 weeks (17-day and

14-week studies) or for 3 to 4 weeks (2-year

studies). 

Stability studies of 4 mg/mL dose formulations (in

deionized water and in 50% ethanol) were conducted

by the analytical chemistry laboratory using highperformance liquid chromatography. Stability was
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

16 Sodium Xylenesulfonate, NTP TR 464 

confirmed for at least 3 weeks when stored in the dark

at room temperature in sealed glass vials and for

3 hours at room temperature open to air and light. 

Homogeneity studies of the doses used in the 2-year

studies and a limited stability study of the 75 mg/mL

dose formulation used in the 2-year studies were

performed by the study laboratory. Homogeneity was

confirmed and the stability of the 75 mg/mL dose

formulation was confirmed for 29 days when stored at

room temperature, and protected from light. 

Periodic analyses of the dose formulations of sodium

xylenesulfonate were conducted by the study

laboratory with high-performance liquid chromatography. During the 17-day studies, doses were

analyzed at the beginning of the study (Table H2). 

During the 14-week studies, doses were analyzed at

the beginning, midpoint, and end of the studies (Table

H3). During the 2-year studies, dose formulations

were analyzed at the beginning of the studies and

every 7 to 10 weeks thereafter (Table H4). Although

the results of a preliminary mixing trial for the 5 and

400 mg/mL concentrations used in the 17-day studies

were within 10% of the target concentrations, the first

set of dose formulations were 12% to 14% greater

than the target concentrations due to a mixing error. 

These formulations were remixed and were all within

10% of the target concentrations (Table H2). Animalroom samples for the 17-day studies were also within

10% of the target concentrations. During the 14-week

studies, all 15 of the dose formulations and all three of

the animal room samples were within 10% of the

target concentrations. In the 2-year studies, all 42 of

the dose formulations and seven of the nine animal

room samples were within 10% of the target

concentrations. For the 14-week studies, two referee

analyses performed by the analytical chemistry

laboratory indicated good agreement with the results

of the study laboratory (Table H5). 

17-DAY STUDIES

Male and female F344/N rats and B6C3F1 mice were

obtained from Taconic Farms (Germantown, NY). 

On receipt, rats were 22 days old, and mice were

31 days old. Animals were quarantined for 12 (rats) 

or 11 (mice) days and were 5 (rats) or 6 (mice) weeks

old on the first day of the studies. Groups of

five male and five female rats and mice received

dermal applications of sodium xylenesulfonate in

distilled water at dose concentrations of 0, 5, 15, 44, 

133, or 400 mg/mL. Doses were applied 5 days per

week for 17 days to the clipped interscapular skin at

a volume of 300 μL for rats and 100 μL for mice

(Appendix I). Feed and water were available

ad libitum. Rats and mice were housed individually. 

Clinical findings for rats and mice were recorded

twice daily and at the end of the studies. Animals

were weighed on days 1 and 8 and at the end of the

studies. Details of the study design and animal

maintenance are summarized in Table 1. 

A necropsy was performed on all animals. The heart, 

right kidney, liver, lungs, right testis, and thymus

were weighed. Histopathologic examination was

restricted to skin from the site of application, skin

from an untreated site, and gross lesions. Table 1 

lists the tissues and organs examined. 

14-WEEK STUDIES

The 14-week studies were conducted to evaluate the

cumulative toxic effects of repeated exposure to

sodium xylenesulfonate and to determine the

appropriate doses to be used in the 2-year studies. 

Male and female F344/N rats and B6C3F1 mice were

obtained from Taconic Farms (Germantown, NY). 

On receipt rats and mice were 31 days old. Animals

were quarantined for 12 (rats) or 19 (mice) days and

were 6 (rats) or 7 (mice) weeks old on the first day of

the studies. Before the studies began, five male and

five female rats and mice were randomly selected for

parasite evaluation and gross observation for evidence

of disease. At the end of the studies, serologic

analyses were performed on five male and five female

rats and mice using the protocols of the NTP Sentinel

Animal Program (Appendix K). 

Groups of 10 male and 10 female rats and mice

received dermal applications of sodium xylenesulfonate in 50% ethanol at dose concentrations of 0, 

5, 15, 44, 133, or 400 mg/mL (Appendix I). A 50% 

solution of ethanol in water was chosen as the vehicle

in the 14-week studies because solutions of sodium

xylenesulfonate in water alone were observed to bead

up rather than to spread out on rodent skin. Doses

were applied 5 days per week for 14 weeks to the

clipped interscapular skin at a volume of 300 μL for

rats and 100 μL for mice. Feed and water were

available ad libitum. Rats and mice were housed
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 17 

individually. Clinical findings for rats and mice were

recorded weekly and at the end of the studies. 

Animals were weighed initially, weekly, and at the

end of the studies. Details of the study design and

animal maintenance are summarized in Table 1. 

For special hematology and clinical chemistry studies, 

additional groups of 10 male and 10 female rats

received dermal applications of sodium xylenesulfonate in 50% ethanol at doses of 0, 5, 15, 44, 

133, or 400 mg/mL. 

Blood was collected from special study rats under

carbon dioxide anesthesia on days 5 and 21 via the

retroorbital sinus. Using the same method, blood was

also collected from core study rats at the end of the

study. For hematology analyses, samples were placed

in containers containing EDTA as an anticoagulant. 

For clinical chemistry analyses, samples were

collected in plastic centrifuge tubes. Hemoglobin

concentrations, hematocrit values, and erythrocyte, 

platelet, and leukocyte counts were measured on an

Ortho ELT-8 analyzer (Ortho Instruments, Westwood, 

MA). Differential leukocyte counts, reticulocyte

counts, and leukocyte, erythrocyte, and platelet

morphologies were determined by light microscopy of

blood films. Mean cell volume, mean cell hemoglobin, and mean cell hemoglobin concentrations were

calculated from the results of analyses for hemoglobin

concentration, hematocrit, and erythrocyte counts. 

The hematology parameters measured are listed in

Table 1. Clinical chemistry parameters were

measured on a Roche Cobas FARA chemistry

analyzer (Roche Diagnostic Systems, Inc., Montclair, 

NJ). The clinical chemistry parameters measured are

listed in Table 1. 

A necropsy was performed on all core study rats and

on all mice. The heart, right kidney, liver, lungs, 

right testis, and thymus were weighed. Tissues for

microscopic examination were fixed and preserved in

10% neutral buffered formalin, processed and

trimmed, embedded in paraffin, sectioned to a

thickness of 5 to 6 μm, and stained with hematoxylin

and eosin. Complete histopathologic examinations

were performed on all control rats (core) and mice

and rats (core) and mice from the 400 mg/mL groups. 

Table 1 lists the tissues and organs routinely

examined. Skin from the site of application was

examined microscopically in progressively lower dose

groups until no skin lesions were observed in a dose

group. 

2-YEAR STUDIES

Groups of 50 male and 50 female rats were

administered dermal applications of 0, 60, 120, or

240 mg sodium xylenesulfonate/kg body weight in

50% ethanol. Groups of 50 male and 50 female mice

were administered dermal applications of 0, 182, 364, 

or 727 mg sodium xylenesulfonate/kg body weight. 

Doses were applied five days per week for 104 (rats

and male mice) or 105 (female mice) weeks to the

clipped interscapular skin at volumes of 85 to 357 μL

for rats and of 46 to 128 μL for mice. Volumes were

adjusted for the weights of the animals throughout the

study (Appendix I). 

Source and Specification of Animals

Male and female F344/N rats and B6C3F1 mice were

obtained from Simonsen Laboratories (Gilroy, CA) 

for use in the 2-year studies. Rats and mice were

quarantined for 14 days prior to the start of the

studies. Five male and five female rats and mice were

selected for parasite evaluation and gross observation

of disease. Serology samples were collected for viral

screening. Rats and mice were approximately

6 weeks old at the beginning of the studies. The

health of the animals was monitored during the studies

according to the protocols of the NTP Sentinel Animal

Program (Appendix K). 

Animal Maintenance

Rats and mice were housed individually. Feed and

water were available ad libitum. Cages were changed

weekly and racks were rotated once every 2 weeks. 

Further details of animal maintenance are given in

Table 1. 

Clinical Examinations and Pathology

All animals were observed twice daily and clinical

findings were recorded monthly. Body weights were

recorded weekly through week 13, monthly thereafter, 

and at the end of the studies. 

All animals were necropsied. A complete histopathologic examination was performed on all rats and mice

that died prior to study termination, and on control

rats and mice, and on 240 mg/kg rats and 727 mg/kg
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

18 Sodium Xylenesulfonate, NTP TR 464 

mice at the end of the studies. Skin from the site

of application was examined for all dose groups. At

necropsy, all organs and tissues were examined for

grossly visible lesions. Major tissues were fixed and

preserved in 10% neutral buffered formalin, 

processed and trimmed, embedded in paraffin, 

sectioned to a thickness of 5 to 6 μm, and stained with

hematoxylin and eosin for microscopic examination. 

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

ovary), samples from each organ were examined. 

Tissues examined microscopically are listed in

Table 1. 

Microscopic evaluations were completed by the study

laboratory pathologist, and the pathology data were

entered into the Toxicology Data Management

System. The slides, paraffin blocks, and residual wet

tissues were sent to the NTP Archives for inventory, 

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

individual animal data records, and pathology tables

were evaluated by an independent quality assessment

laboratory. The individual animal records and tables

were compared for accuracy, the slide and tissue

counts were verified, and the histotechnique was

evaluated. For the 2-year studies, a quality

assessment pathologist reviewed the skin of rats and

mice and the liver of mice for all lesions; the liver and

spleen of male rats for mononuclear cell leukemia; 

and the clitoral gland of female rats for proliferative

lesions (hyperplasia, adenoma, and carcinoma). 

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 diagnoses

made by the laboratory and quality assessment

pathologists. Representative histopathology slides

containing examples of lesions related to chemical

administration, examples of disagreements in

diagnoses between the laboratory and quality

assessment pathologist, or lesions of general interest

were presented by the chairperson to the PWG for

review. The PWG consisted of the quality assessment

pathologist and other pathologists experienced in

rodent toxicologic pathology. This group examined

the tissues without any knowledge of dose groups or

previously rendered diagnoses. When the PWG

consensus differed from the opinion of the laboratory

pathologist, the diagnosis was changed. Final

diagnoses for reviewed lesions represent a consensus

between the laboratory pathologist, reviewing

pathologist(s), and the PWG. The PWG reviewed the

following: proliferative lesions of the thyroid gland in

all female mice and in control and 727 mg/kg male

mice; all control male rat testes for interstitial cell

neoplasms; clitoral glands in female rats and preputial

glands in control and 240 mg/kg male rats for

proliferative lesions; thyroid, adrenal (medulla), and

pituitary glands in control and 240 mg/kg groups of

male and female rats. Details of these review

procedures have been described, in part, by Maronpot

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

subsequent analyses of the pathology data, the

decision of whether to evaluate the diagnosed lesions

for each tissue type separately or combined was

generally based on the guidelines of McConnell et al. 

(1986). 

STATISTICAL METHODS

Survival Analyses

The probability of survival was estimated by the

product-limit procedure of Kaplan and Meier (1958) 

and is presented in the form of graphs. Animals

found dead of other than natural causes or missing

were censored from the survival analyses; animals

dying from natural causes were not censored. 

Statistical analyses for 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, A4, B1, B4, C1, C5, D1, 

and D5 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 most neoplasms (Tables A3, B3, C3, 

and D3) and all nonneoplastic lesions are given as the

numbers of animals affected at each site examined

microscopically. However, when macroscopic

examination was required to detect neoplasms in

certain tissues (e.g., skin, intestine, harderian gland, 

and mammary gland) before microscopic evaluation, 

or when neoplasms had multiple potential sites of

occurrence (e.g., leukemia or lymphoma), the

denominators consist of the number of animals on
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 19 

which a necropsy was performed. Tables A3, B3, 

C3, and D3 also give the survival-adjusted neoplasm

rate for each group and each site-specific neoplasm, 

i.e., the Kaplan-Meier estimate of the neoplasm

incidence that would have been observed at the end of

the study in the absence of mortality from all

competing risks (Kaplan and Meier, 1958) 

Analysis of Neoplasm Incidences

The majority of neoplasms in these studies were

considered to be incidental to the cause of death or not

rapidly lethal. Thus, the primary statistical method

used was logistic regression analysis, which assumed

that the diagnosed neoplasms were discovered as the

result of death from an unrelated cause and thus did

not affect the risk of death. In this approach, 

neoplasm prevalence was modeled as a logistic

function of chemical exposure and time. Both linear

and quadratic terms in time were incorporated

initially, and the quadratic term was eliminated if the

fit of the model was not significantly enhanced. The

neoplasm incidences of exposed and control groups

were compared on the basis of the likelihood score

test for the regression coefficient of dose. This

method of adjusting for intercurrent mortality is the

prevalence analysis of Dinse and Lagakos (1983), 

further described and illustrated by Dinse and

Haseman (1986). When neoplasms are incidental, this

comparison of the time-specific neoplasm prevalences

also provides a comparison of the time-specific

neoplasm incidences (McKnight and Crowley, 1984). 

In addition to logistic regression, other methods of

statistical analysis were used, and the results of these

tests are summarized in the appendixes. These

methods include the life table test (Cox, 1972; 

Tarone, 1975), appropriate for rapidly lethal

neoplasms, and the Fisher exact test and the CochranArmitage trend test (Armitage, 1971; Gart et al., 

1979), procedures based on the overall proportion of

neoplasm-bearing animals. 

Tests of significance included pairwise comparisons of

each exposed group with controls and a test for an

overall dose-related trend. Continuity-corrected tests

were used in the analysis of neoplasm incidence, and

reported P values are one sided. The procedures

described in the preceding paragraphs were also used

to evaluate selected nonneoplastic lesions. For further

discussion of these statistical methods, refer to

Haseman (1984). 

Analysis of Nonneoplastic Lesion Incidences

Because all nonneoplastic lesions in this study were

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 nonneoplastic

lesion prevalence was modeled as a logistic function

of chemical exposure and time. For lesions detected

at 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

and control groups in the analysis of continuous

variables. Organ and body weight data, which have

approximately normal distributions, were analyzed

using the parametric multiple comparison procedures

of Dunnett (1955) and Williams (1971, 1972). 

Clinical chemistry and hematology data, which have

typically skewed distributions, were analyzed using

the nonparametric multiple comparison methods of

Shirley (1977) and Dunn (1964). Jonckheere’s test

(Jonckheere, 1954) was used to 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 identified by the outlier test of Dixon and

Massey (1951) were examined by NTP personnel, and

implausible values were eliminated from the analysis. 

Average severity values were analyzed for

significance using the Mann-Whitney U test

(Hollander and Wolfe, 1973). 

Historical Control Data

Although the concurrent control group is always the

first and most appropriate control group used for

evaluation, historical control data can be helpful in the

overall assessment of neoplasm incidence in certain

instances. Consequently, neoplasm incidences from

the NTP historical control database, which is updated

yearly, are included in the NTP reports for neoplasms

appearing to show compound-related effects. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

20 Sodium Xylenesulfonate, NTP TR 464 

QUALITY ASSURANCE METHODS

The 14-week and 2-year studies were conducted in

compliance with Food and Drug Administration Good

Laboratory Practice Regulations (21 CFR, Part 58). 

In addition, as records from the 2-year studies were

submitted to the NTP Archives, these studies were

audited retrospectively by an independent 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 the

reports and are on file at NIEHS. The audit findings

were reviewed and assessed by NTP staff, so all

comments had been resolved or were otherwise

addressed during the preparation of this Technical

Report. 

GENETIC TOXICOLOGY

The genetic toxicity of sodium xylenesulfonate was

assessed by testing the ability of the chemical to

induce mutations in various strains of Salmonella

typhimurium, sister chromatid exchanges and

chromosomal aberrations in cultured Chinese hamster

ovary cells, and mutations in L5178Y mouse

lymphoma cells. The protocols for these studies and

the results are given in Appendix E. 

The genetic toxicity studies of sodium xylenesulfonate

are part of a larger effort by the NTP to develop a

database that would permit the evaluation of

carcinogenicity in experimental animals from the

structure and responses of the chemical in short-term

in vitro and in vivo genetic toxicity tests. These

genetic toxicity tests were originally developed to

study mechanisms of chemically induced DNA

damage and to predict carcinogenicity in animals, 

based on the electrophilic theory of chemical

carcinogenesis and the somatic mutation theory

(Miller and Miller, 1977; Straus, 1981; Crawford, 

1985). 

There is a strong correlation between a chemical’s

potential electrophilicity (structural alert to DNA

reactivity), mutagenicity in Salmonella, and carcinogenicity in rodents. The combination of electrophilicity and Salmonella mutagenicity is highly

correlated with the induction of carcinogenicity in rats

and mice and/or at multiple tissue sites (Ashby and

Tennant, 1991). Other in vitro genetic toxicity tests

do not correlate well with rodent carcinogenicity

(Tennant et al., 1987; Zeiger et al., 1990), although

these other tests can provide information on the types

of DNA and chromosome effects that can be induced

by the chemical being investigated. Data from NTP

studies show that a positive response in Salmonella is

currently the most predictive in vitro test for rodent

carcinogenicity (89% of the Salmonella mutagens

were rodent carcinogens), and that there is no

complementarity among the in vitro genetic toxicity

tests. That is, no battery of tests that included the

Salmonella test improved the predictivity of the

Salmonella test alone. 
 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 21 

TABLE 1 

Experimental Design and Materials and Methods in the Dermal Studies of Sodium Xylenesulfonate

17-Day Studies 14-Week Studies 2-Year Studies

Study Laboratory

Southern Research Institute Southern Research Institute Battelle Columbus Laboratories

(Birmingham, AL) (Birmingham, AL) (Columbus, OH) 

Strain and Species

Rats: F344/N Rats: F344/N Rats: F344/N

Mice: B6C3F1 Mice: B6C3F1 Mice: B6C3F1 

Animal Source

Taconic Farms Taconic Farms Simonsen Laboratories, Inc. 

(Germantown, NY) (Germantown, NY) (Gilroy, CA) 

Time Held Before Studies

Rats: 12 days Rats: 12 days 14 days

Mice: 11 days Mice: 19 days

Average Age When Studies Began

Rats: 5 weeks Rats: 6 weeks 7 weeks

Mice: 6 weeks Mice: 7 weeks

Date of First Dose

Rats: 20 July 1987 Rats: 16 February 1988 Rats: 29 November 1990 

Mice: 27 July 1987 Mice: 23 February 1988 Mice: 20 December 1990 

Duration of Dosing

5 days per week for 17 days 5 days per week for 14 weeks 5 days per week for 104 weeks (rats and

male mice) or 105 weeks (female mice) 

Date of Last Dose

Rats: 5-6 August 1987 Rats: 17-19 May 1988 (core) Rats: 20 November 1992 

Mice: 11-12 August 1987 16-17 May 1988 (special study) Mice: 11 December 1992 (males) 

Mice: 24-26 May 1988 18 December 1992 (females) 

Necropsy Dates

Rats: 5-6 August 1987 Rats: 18-20 May 1988 Rats: 2-3 December 1992 

Mice: 12-13 August 1987 Mice: 25-27 May 1988 Mice: 21-23 December 1992 (males) 

28-30 December 1992 (females) 

Average Age at Necropsy

Rats: 7 weeks Rats: 19-20 weeks Rats: 111-112 weeks

Mice: 8 weeks Mice: 20-21 weeks Mice: 111-112 weeks
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

22 Sodium Xylenesulfonate, NTP TR 464 

TABLE 1
#

Experimental Design and Materials and Methods in the Dermal Studies of Sodium Xylenesulfonate (continued)
#

17-Day Studies 14-Week Studies 2-Year Studies

Size of Study Groups

5 males and 5 females 10 males and 10 females (rats and 50 males and 50 females

mice; core study) 

10 male and 10 female rats (special

study) 

Method of Distribution

Animals were distributed randomly into Same as 17-day studies Same as 17-day studies

groups of approximately equal initial

mean body weights. 

Animals per Cage

1 1 1 

Method of Animal Identification

Toe clip Toe clip Tail tattoo

Diet

NIH-07 open formula pelleted diet Same as 17-day studies Same as 17-day studies

(Zeigler Brothers, Inc., Gardners, PA), 

available ad libitum

Water Distribution

Tap water (City of Birmingham Same as 17-day studies Tap water (Columbus municipal supply) 

municipal supply) via automatic via automatic watering system (Edstrom

watering system (Edstrom Industries, Industries, Waterford, WI), available ad

Waterford, WI), available ad libitum libitum

Cages

Polycarbonate (Lab Products Inc., Same as 17-day studies Same as 17-day studies

Maywood, NJ), changed once weekly

Bedding

Heat-treated hardwood chips Same as 17-day studies Sani-Chip® hardwood chips

(P.J. Murphy Forest Products Co., (P.J. Murphy Forest Products Corp., 

Montville, NJ) Montville, NJ) 

Cage Filters

Reemay® spun-bonded polyester Same as 17-day studies DuPont 2024 spun-bonded polyester

(Andico, Birmingham, AL) (Snow Filtration Co., Cincinnati, OH) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 23 

TABLE 1
#

Experimental Design and Materials and Methods in the Dermal Studies of Sodium Xylenesulfonate (continued)
#

17-Day Studies 14-Week Studies 2-Year Studies

Racks

Stainless steel (Lab Products Inc., Stainless steel (Lab Products Inc., Stainless steel (Lab Products Inc., 

Maywood, NJ), not rotated Maywood, NJ), rotated once every Maywood, NJ), rotated once every

2 weeks 2 weeks

Animal Room Environment

Temperature: 22.5E to 25.6E C (rats); Temperature: 20.1E to 24.5E C (rats); Temperature: 20.0E to 24.4E C (rats); 

15.8E to 23.9E C (mice) 21.5E to 23.9E C (mice) 16.7E to 25.6E C (mice) 

Relative humidity: 51% to 68% (rats); Relative humidity: 20% to 67% (rats); Relative humidity: 35% to 70% (rats); 

31% to 50% (mice) 15% to 63% (mice) 34% to 69% (mice) 

Fluorescent light: 12 hours/day Fluorescent light: 12 hours/day Fluorescent light: 12 hours/day

Room air: minimum of Room air: minimum of Room air: minimum of

10 changes/hour 10 changes/hour 10 changes/hour

Doses

0, 5, 15, 44, 133, or 400 mg/mL 0, 5, 15, 44, 133, or 400 mg/mL Rats: 0, 60, 120, or 240 mg/kg

(dose volumes of 300 μL for rats and (dose volumes of 300 μL for rats and (dose volumes of 85 to 357 μL) 

100 μL for mice) 100 μL for mice) Mice: 0, 182, 364, or 727 mg/kg (dose

volumes of 46 to 128 μL) 

Type and Frequency of Observation

Observed twice daily; animals were Observed twice daily; animals were Observed twice daily; clinical findings

weighed on days 1 and 8, and at the end weighed initially, weekly, and at the end were recorded monthly, body weights

of the studies. Clinical findings were of the studies. Clinical findings were were recorded weekly through week 13, 

recorded twice daily and at the end of recorded weekly and at the end of the monthly thereafter, and at the end of the

the studies. studies. Feed consumption was studies. 

measured weekly for each animal. 

Method of Sacrifice

CO2 asphyxiation CO2 asphyxiation CO2 asphyxiation

Necropsy

Necropsy performed on all animals. Necropsy performed on all core study Necropsy performed on all animals. 

Organs weighed included the heart, right rats and on all mice. Organs weighed

kidney, liver, lungs, right testis, and included the heart, right kidney, liver, 

thymus. lungs, right testis, and thymus. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

24 Sodium Xylenesulfonate, NTP TR 464 

TABLE 1
#

Experimental Design and Materials and Methods in the Dermal Studies of Sodium Xylenesulfonate (continued)
#

17-Day Studies 14-Week Studies 2-Year Studies

Clinical Pathology

None Blood was collected from special study None

rats on day 5 and on day 21 and from

core study rats at the end of the study. 

Blood was collected from the

retroorbital sinus. 

Hematology: hematocrit, hemoglobin

concentration, erythrocyte and

reticulocyte counts, mean cell volume, 

mean cell hemoglobin, mean cell

hemoglobin concentration, platelet

count, and leukocyte count and

differentials. 

Clinical Chemistry: urea nitrogen, 

creatinine, total protein, albumin, 

alanine aminotransferase, alkaline

phosphatase, creatine kinase, sorbitol

dehydrogenase, and bile acids. 

Histopathology

Histopathologic examination was limited Complete histopathologic examinations Complete histopathologic examinations

to the skin (site of application and were performed on control rats (core) were performed on all control, on all

untreated skin) and gross lesions. and mice, and rats (core study) and mice 240 mg/kg rats and 727 mg/kg mice, 

from the 400 mg/mL groups. In and on all animals that died early. In

addition to gross lesions, tissue masses, these animals, in addition to gross

and associated lymph nodes, the tissues lesions, tissue masses, and associated

examined included: adrenal gland, bone lymph nodes, the tissues examined

and marrow, brain, clitoral gland, included: adrenal gland, bone and

esophagus, gallbladder (mice), heart, marrow, brain, clitoral gland, 

kidney, large intestine (cecum, colon, esophagus, gallbladder (mice), heart, 

and rectum), small intestine (duodenum, kidney, large intestine (cecum, colon, 

jejunum, and ileum), liver, lung, lymph and rectum), small intestine (duodenum, 

nodes (mandibular and mesenteric), jejunum, and ileum), liver, lung, lymph

mammary gland, nose, ovary, pancreas, nodes (mandibular and mesenteric), 

parathyroid gland, pituitary gland, mammary gland, nose, ovary, pancreas, 

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

gland, skin, spleen, stomach preputial gland, prostate gland, salivary

(forestomach and glandular stomach), gland, spleen, stomach (forestomach and

testis with epididymis and seminal glandular stomach), testis with

vesicle, thymus, thyroid gland, trachea, epididymis and seminal vesicle, thymus, 

urinary bladder, and uterus. Skin thyroid gland, trachea, urinary bladder, 

samples were examined in lower dose and uterus. In 60 and 120 mg/kg rats

groups until a no-effect level was and in 182 and 364 mg/kg mice, tissues

reached. examined microscopically included skin

from the site of application and control

skin. Additional review was made of the

liver and spleen of 60 and 120 mg/kg

male rats for mononuclear cell leukemia; 

the clitoral gland of female rats and the

preputial gland of control and high-dose

male rats for proliferative lesions; the

liver of 182 and 364 mg/kg male and

female mice; and the thyroid gland of

182 and 364 mg/kg female mice. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

25 

RESULTS
 

RATS

17-DAY STUDY

All rats survived to the end of the study (Table 2). 

Final mean body weights and body weight gains of

dosed rats were similar to those of the control groups. 

Dermal applications of 300 μL of 5, 15, 44, 133, and

400 mg/mL delivered average daily doses of

approximately 10, 30, 90, 260, and 800 mg sodium

xylenesulfonate/kg body weight to males and 13, 40, 

120, 330, and 1,030 mg/kg to females. Clinical

findings generally involved the skin of dosed animals

and included tan or brown skin discoloration and

crusty white deposits (presumed to be dried chemical) 

at the site of application. Neither of these observations

were considered significant findings. The relative

liver weights of 133 and 400 mg/mL male and female

rats were significantly greater than those of the control

groups, but the absolute weights were similar. The

biological significance of the differences in relative

liver weights was unclear (Table F1). In males and

females, the few skin lesions observed grossly and

microscopically were generally attributed to repeated

clipping and were not considered related to chemical

administration. 

TABLE 2 

Survival and Body Weights of Rats in the 17-Day Dermal Study of Sodium Xylenesulfonate

Mean Body Weightb (g) Final Weight a Dose Survival Initial Final Change Relative to Controls

(mg/mL) (%) 

Male

0 5/5 110 ± 5 181 ± 7 71 ± 1 

5 5/5 111 ± 6 188 ± 6 78 ± 1 104 

15 5/5 112 ± 5 187 ± 8 75 ± 3 103 

44 5/5 106 ± 4 174 ± 7 68 ± 4 96 

133 5/5 114 ± 6 188 ± 12 74 ± 7 104 

400 5/5 111 ± 5 186 ± 8 75 ± 4 102 

Female

0 5/5 92 ± 6 136 ± 5 44 ± 3 

5 5/5 97 ± 5 137 ± 5 40 ± 1 101 

15 5/5 98 ± 5 137 ± 5 39 ± 2 101 

44 5/5 93 ± 2 133 ± 4 40 ± 3 98 

133 5/5 100 ± 3 139 ± 5 40 ± 4 103 

400 5/5 98 ± 3 134 ± 5 36 ± 3 99 

a Number of animals surviving at 17 days/number initially in group b Weights and weight changes are given as mean ± standard error. Differences from the control group were not significant by Dunnett’s test. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

26 Sodium Xylenesulfonate, NTP TR 464 

14-WEEK STUDY

All rats survived to the end of the study (Table 3). 

Final mean body weights and body weight gains of

dosed male and female rats were similar to those of

the control groups. Dermal applications of 300 μL of

5, 15, 44, 133, and 400 mg/mL delivered average

daily doses of approximately 6, 20, 60, 170, and

500 mg sodium xylenesulfonate/kg body weight to

males and 10, 30, 90, 260, and 800 mg/kg to females. 

The only notable clinical finding was brown

discoloration of the skin at the site of application in

dosed animals. 

Hematology and clinical chemistry parameters of

dosed groups of males and females were significantly

different from those of the controls in several

instances, but these differences were sporadic and did

not demonstrate a treatment relationship

(Appendix G). 

The absolute and relative liver weights of males

receiving 44, 133, or 400 mg/mL were significantly

less than those of the control group (Table F2). There

were no treatment-related histopathologic alterations

in the livers of dosed male and female rats; thus, the

biological significance of the decreased liver weights

was unclear. 

Minimal hyperplasia of the epidermis at the site of

application occurred in male and female rats from the

control groups as well as most dosed groups. The

incidence of epidermal hyperplasia in 400 mg/mL

males was considered to be possibly chemical-related

(Table 4). Lesions in rats from lower dose groups

were attributed to vehicle application and weekly

clipping. Epidermal hyperplasia typically consisted of

a minimal multifocal increase in the thickness of the

epidermis. 

TABLE 3 

Survival and Body Weights of Rats in the 14-Week Dermal Study of Sodium Xylenesulfonate

Mean Body Weightb (g) Final Weight

Dose

(mg/mL) 

Survivala Initial Final Change Relative to Controls

(%) 

Male

0 

5 

15 

44 

133 

400 

10/10 

10/10 

10/10 

10/10 

10/10 

10/10 

147 ± 5 

145 ± 6 

136 ± 5 

133 ± 7 

141 ± 4 

144 ± 7 

350 ± 8 

346 ± 5 

346 ± 6 

328 ± 6 

339 ± 8 

344 ± 5 

203 ± 8 

202 ± 4 

209 ± 5 

195 ± 9 

199 ± 8 

200 ± 6 

99 

99 

94 

97 

98 

Female

0 

5 

15 

44 

133 

400 

10/10 

10/10 

10/10 

10/10 

10/10 

10/10 

110 ± 3 

103 ± 3 

102 ± 3 

99 ± 3* 

107 ± 2 

107 ± 2 

195 ± 4 

189 ± 3 

188 ± 2 

190 ± 2 

197 ± 4 

195 ± 3 

85 ± 3 

86 ± 3 

86 ± 3 

90 ± 3 

90 ± 3 

88 ± 3 

97 

96 

98 

101 

100 

* Significantly different (P#0.05) from the control group by Dunnett’s test a Number of animals surviving at 14 weeks/number initially in group b Weights and weight changes are given as mean ± standard error. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

27 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE 4 

Incidences of Epidermal Hyperplasia in Rats in the 14-Week Dermal Study

of Sodium Xylenesulfonate

Vehicle 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

Control

Male

Skin, Site of Applicationa

Epidermal Hyperplasiab

10 

1 (1.0) c 10 3 (1.0) 

10 

6* (1.0) 

10 

0 

10 

0 

10 

8** (1.0) 

Female

Skin, Site of Application

Epidermal Hyperplasia

10 

5 (1.0) 

10 

3 (1.0) 

10 

1 (1.0) 

10 

6 (1.0) 

10 

6 (1.0) 

10 

3 (1.0) 

* Significantly different (P#0.05) from the vehicle control by the Fisher exact test

** P#0.01 a Number of rats with skin examined microscopically b Number of rats with lesion
 

Average severity grade of lesion in affected animals: 1  ̆ minimal; 2  ̆ mild; 3  ̆ moderate; 4  ̆ marked
 

Dose Selection Rationale: In the 17-day and 14-week deliver the 240 mg/kg dose) in the 2-year study. 

studies, dose concentrations of up to 400 mg/mL were Fixed volumes and concentrations were used in the

well tolerated. The only limitation to the 17-day and 14-week studies which resulted in

concentration used in dermal dosing in the 2-year decreasing dose concentrations (mg/kg body weight) 

study was the difficulty of getting a uniform as the animals grew (Appendix I). Constant dose

suspension of sodium xylenesulfonate in the vehicle. concentrations were achieved in the 2-year study by

To obtain uniform suspensions, the highest dose adjusting the applied dose volumes to the mean body

concentration was decreased to 300 mg/mL (used to weights throughout the study. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

28 Sodium Xylenesulfonate, NTP TR 464 

2-YEAR STUDY

Survival

Estimates of 2-year survival probabilities for male and

female rats are shown in Table 5 and in the KaplanMeier survival curves (Figure 1). Survival of dosed

males and females was similar to that of the control

groups and consistent with survival of control groups

in other NTP studies in which rats were individually

housed (Rao, 1995). 

Body Weights and Clinical Findings

Mean body weights of dosed males and females

were similar to those of the controls throughout the

study. (Tables 6 and 7, Figure 2). In males, there

were no clinical findings considered treatment related. 

In females, clinical findings were limited to irritation

at the site of application in one control female, four

120 mg/kg females, and two 240 mg/kg females. 

TABLE 5 

Survival of Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Male

Animals initially in study	 50 50 50 50 

Moribund 28 25 22 30 

Natural deaths 15 8 19 10 

Animals surviving to study termination 7 17 9 10 a Percent probability of survival at end of study 14 34 18 20 

Mean survival (days)

b 638 634 632 635 

c Survival analysis	 P ̆1.000N P ̆0.115N P ̆0.927N P ̆0.580N

Female

Animals initially in study	 50 50 50 50 

d Accidental death 1 0 0 0 

Moribund 14 10 15 12 

Natural deaths 13 24 18 22 e Animals surviving to study termination 22 16 17 16 

Percent probability of survival at end of study 46 32 34 32 

Mean survival (days) 647 610 641 621 

Survival analysis	 P ̆0.309 P ̆0.156 P ̆0.329 P ̆0.193 

a Kaplan-Meier determinations

b	 Mean of all deaths (uncensored, censored, and terminal sacrifice) 

The result of the life table trend test (Tarone, 1975) is in the vehicle control column, and the results of the life table pairwise comparisons

(Cox, 1972) with the vehicle controls are in the dosed columns. A negative trend or lower mortality in a dose group is indicated by N. d	 Censored from survival analyses e Includes one animal that died during the last week of the study

c
 

Sodium Xylenesulfonate, NTP TR 464 29 

FIGURE 1
 

Kaplan-Meier Survival Curves for Male and Female Rats
 

Administered Sodium Xylenesulfonate Dermally for 2 Years
 
 

 

30 Sodium Xylenesulfonate, NTP TR 464 

TABLE 6 

Mean Body Weights and Survival of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Weeks Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

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

1 127 50 127 100 50 128 100 50 127 100 50 

2 155 50 154 100 50 154 99 50 154 100 50 

3 187 50 188 101 50 185 99 50 187 100 50 

4 206 50 208 101 50 206 100 50 206 100 50 

5 228 50 231 101 50 228 100 50 227 99 50 

6 245 50 245 100 50 243 99 50 241 99 50 

7 259 50 261 101 50 259 100 50 256 99 50 

8 271 50 271 100 50 270 100 50 265 98 50 

9 282 50 282 100 50 280 99 50 277 98 50 

10 292 50 293 100 50 290 99 50 287 98 50 

11 298 50 298 100 50 295 99 50 292 98 50 

12 305 50 307 101 50 304 100 50 299 98 50 

13 311 50 312 100 50 309 99 50 305 98 50 

17 336 50 335 100 50 329 98 50 327 97 50 

21 356 50 352 99 50 347 97 50 341 96 50 

25 369 50 364 99 50 360 98 50 354 96 50 

29 382 50 380 99 49 374 98 50 368 96 50 

33 393 50 391 100 49 384 98 50 382 97 50 

37 399 50 397 100 49 389 97 50 387 97 50 

41 407 50 405 100 49 400 98 49 392 96 50 

45 414 50 414 100 49 411 99 49 402 97 50 

49 414 50 415 100 49 412 99 49 407 98 50 

52 421 50 420 100 47 415 99 49 412 98 50 

57 432 50 428 99 47 427 99 49 422 98 49 

61 433 50 436 101 46 433 100 49 426 98 49 

65 429 48 437 102 45 431 101 48 429 100 47 

69 433 48 442 102 45 432 100 47 432 100 47 

73 439 46 437 99 44 434 99 45 429 98 46 

77 434 45 440 102 42 436 101 44 432 100 41 

81 438 39 446 102 39 432 99 42 427 98 37 

85 431 35 444 103 38 426 99 37 424 98 33 

89 427 30 438 103 33 423 99 32 417 98 30 

93 416 26 424 102 27 413 99 27 416 100 26 

97 410 21 414 101 24 419 102 18 409 100 22 

101 376 16 390 104 23 373 99 15 397 106 20 

104 376 9 372 99 18 358 95 9 366 97 13 

Mean for weeks

1-13 244 244 100 242 99 240 98 

14-52 389 387 99 382 98 377 97 

53-104 421 427 101 418 99 417 99 
 

 

Sodium Xylenesulfonate, NTP TR 464 31 

TABLE 7 

Mean Body Weights and Survival of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Weeks Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

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

1 108 50 107 100 50 107 99 50 107 100 50 

2 124 50 124 100 50 124 100 50 124 100 50 

3 136 50 136 100 50 137 101 50 135 100 50 

4 144 50 143 99 50 144 100 50 142 99 50 

5 149 50 149 100 50 150 100 50 147 98 50 

6 158 50 156 99 50 159 101 50 155 98 50 

7 163 50 162 100 50 164 101 50 161 99 50 

8 165 50 164 100 50 167 101 50 162 98 50 

9 171 50 170 99 50 172 100 50 167 98 50 

10 174 50 173 99 50 174 100 50 170 98 50 

11 177 50 176 99 50 177 100 50 174 98 50 

12 179 50 179 100 50 180 100 50 176 98 50 

13 182 50 181 99 50 182 100 50 179 98 50 

17 189 50 188 100 50 190 101 50 186 98 50 

21 196 50 195 100 50 197 100 50 191 98 50 

25 201 50 200 100 50 202 101 50 196 98 50 

29 210 50 208 99 50 210 100 50 205 98 50 

33 216 50 215 100 49 217 100 50 211 98 50 

37 220 50 219 99 49 221 101 50 213 97 50 

41 227 50 226 100 48 226 99 50 218 96 50 

45 234 49 232 99 47 233 100 50 225 96 49 

49 240 49 238 99 47 239 100 50 228 95 48 

52 246 49 246 100 47 244 99 50 235 96 47 

57 255 48 256 100 45 253 99 50 246 96 45 

61 262 45 261 99 45 259 99 50 252 96 43 

65 263 45 263 100 43 260 99 47 253 96 42 

69 264 45 266 101 43 264 100 46 255 97 42 

73 266 40 266 100 41 266 100 44 254 95 42 

77 271 40 268 99 38 272 100 40 262 97 38 

81 273 38 275 101 33 273 100 37 266 98 37 

85 274 37 277 101 29 272 99 35 266 97 35 

89 279 35 276 99 29 273 98 32 268 96 31 

93 282 33 288 102 22 281 100 28 271 96 27 

97 282 31 283 100 21 290 103 23 268 95 24 

101 289 26 278 96 19 285 98 22 270 93 20 

104 277 23 278 101 17 282 102 17 271 98 16 

Mean for weeks

1-13 156 155 99 157 101 154 99 

14-52 218 217 100 218 100 211 97 

53-104 272 272 100 272 100 262 96 
 

32 Sodium Xylenesulfonate, NTP TR 464 

FIGURE 2
 

Growth Curves for Male and Female Rats Administered
 

Sodium Xylenesulfonate Dermally for 2 Years
 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 33 

Pathology and Statistical Analysis

This section describes the statistically significant and

biologically noteworthy changes in the incidences of

mononuclear cell leukemia and neoplasms and

nonneoplastic lesions of the skin and other organs. 

Summaries of the incidences of neoplasms and

nonneoplastic lesions, individual animal tumor

diagnoses, and statistical analyses of primary

neoplasms that occurred with incidences of at least

5% in at least one animal group are presented in

Appendix A for male rats and Appendix B for female

rats. 

Skin: Incidences of neoplasms in the skin and

subcutaneous tissues of male rats are given in

Tables 8, A1, and A3. The incidences and morphologies of these neoplasms were typical of spontaneous

neoplasms in F344/N rats and, therefore, were not

considered related to treatment. There were no skin

neoplasms observed at the site of application in female

rats. 

Hyperplasia of the epidermis at the site of application

occurred at low incidences in males in the 60, 120, 

and 240 mg/kg groups (Tables 8 and A4). 

Hyperplasia of the epidermis at the site of application

occurred at low incidences in females in the 120 and

240 mg/kg groups and occurred with a significant

positive trend (P#0.05; Tables 8 and B4). Incidences

of hyperplasia of the epidermis might have been

related to chemical administration. These multifocal

lesions were minimal to mild in severity and consisted

of extensive increased numbers of cell layers of the

epidermal epithelium. 

Other nonneoplastic lesions of the skin did not appear

related to chemical administration, but more probably

were a result of repeated hair clipping and vehicle

application. Focal ulceration of the epidermis

occurred in control, 120 mg/kg, and 240 mg/kg

females (Tables 8 and B4). Ulceration through the

layers of the epidermal epithelium was typically

accompanied by inflammatory cells in the exposed

dermis and an effluxing of protein and necrotic cells

which were diagnosed as exudate. Hyperplasia of the

sebaceous glands occurred at low incidences in control

and 60 mg/kg males and in control, 120 mg/kg, and

240 mg/kg females (Tables 8, A4, and B4) and

consisted of numerous large sebaceous glands in the

dermis. Inflammation of the dermis or subcutaneous

tissues consisted of accumulations of both mononuclear and neutrophil inflammatory cells. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

34 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE 8 

Incidences of Neoplasms and Nonneoplastic Lesions of the Skin in Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Male

Skin, Site of Applicationa

Epidermis, Hyperplasiab

Sebaceous Gland, Hyperplasia

Inflammation, Chronic Active

50 

0 

1 

0 

(2.0) 

50 

1 

1 

1 

(1.0)c

(1.0) 

(1.0) 

50 

1 

0 

0 

(1.0) 

50 

2 

0 

0 

(1.5) 

Sebaceous Gland Carcinoma

Basal Cell Adenoma

Subcutaneous Tissue, Fibroma, Multiple

Subcutaneous Tissue, Fibrous Histiocytoma

1 

0 

1 

0 

0 

1 

0 

1 

0 

1 

0 

0 

0 

0 

0 

0 

Female

Skin, Site of Application

Epidermis, Exudate

Epidermis, Hyperplasia

Epidermis, Inflammation, Chronic Active

Epidermis, Ulcer

Sebaceous Gland, Hyperplasia

Inflammation, Chronic Active

50 

0 

1 

0 

1 

2 

1 

(2.0) 

(3.0) 

(1.5) 

(2.0) 

50 

0 

0 

0 

0 

0 

0 

50 

0 

4 

0 

2 

2 

2 

(1.8) 

(3.0) 

(2.5) 

(3.0) 

50 

2 

5 

1 

2 

2 

2 

(1.0) 

(1.4) 

(1.0) 

(3.0) 

(2.0) 

(2.5) 

a Number of rats with skin examined microscopically b Number of rats with lesion
 

Average severity grade of lesions in affected animals: 1  ̆ minimal; 2  ̆ mild; 3  ̆ moderate; 4  ̆ marked
 

Other Organs: There was no relation to dose in the All Organs: Mononuclear cell leukemia is a common

incidences of pituitary gland adenoma in male rats neoplasm in F344/N rats in 2-year studies. The

(44/50, 27/33, 34/41, 36/50; Table A3). The control incidences of mononuclear cell leukemia in 60 and

incidence was unusually high compared to historical 240 mg/kg males were significantly greater than in the

control incidences from some dermal and feed studies control group by the logistic regression test (0 mg/kg, 

[dermal acetone: 59/100 (59.0% ± 15.6%); range, 12/50; 60 mg/kg, 24/50; 120 mg/kg, 15/50; 

48%-70%; dermal ethanol: 31/52; feed: 377/1,284 240 mg/kg, 25/50; Table A3) but not by the life table

(29.4% ± 10.6%); range, 14%-60%]; however, it test (a more appropriate statistic for this generally

was similar to controls in a “neat” dermal study fatal neoplasm). The incidence in the male control

(39/44). Likewise, the incidences of testicular group was low compared to the mean incidences in

interstitial cell adenoma were not treatment related NTP historical controls for two dermal studies

(13/50, 6/33, 8/41, 13/50; Table A1), but the control [dermal acetone: 40/100 (40.0% ± 11.3%); range, 

incidences were lower than historical control 32%-48%; dermal ethanol: 23/52] although higher

incidences from dermal and feed studies [dermal than that of the control group in a “neat” dermal study

acetone: 75/100 (75.0% ± 18.4%); range, 62%-88%; (9/50). The marginal increases in mononuclear cell

dermal ethanol: 42/52; feed: 1,169/1,302 leukemia in rats were considered unrelated to

(89.8% ± 5.9%); range, 74%-98%] yet higher than chemical administration because of the lack of

controls in a “neat” dermal study (4/50). The statistical significance by the life table analysis and the

biological significance of increased incidences of lack of a clear dose-response relationship. 

pituitary gland adenoma and decreased incidences of

testicular interstitial cell adenoma in controls was

unclear. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 35 

MICE

17-DAY STUDY

All mice survived to the end of the study (Table 9). 

Final mean body weights and body weight gains of

dosed mice were similar to those of the controls. 

Dermal applications of 100 μL of 5, 15, 44, 133, and

400 mg/mL delivered average daily doses of approximately 20, 60, 190, 540, and 1,600 mg sodium

xylenesulfonate/kg body weight to males and 26, 80, 

220, 680, and 2,000 mg/kg to females. Clinical

findings included crusty white deposits (presumed to

TABLE 9 

be dried chemical) at the site of application in two

133 mg/mL males and in all 400 mg/mL males and

females. The absolute and relative liver weights of 15 

and 44 mg/mL males and 400 mg/mL males and

females were significantly greater than those of the

control groups, but the biological significance of these

differences was unclear (Table F3). The few skin

lesions observed grossly and microscopically in males

and females were generally attributed to repeated

clipping and were not considered related to chemical

administration. 

Survival and Body Weights of Mice in the 17-Day Dermal Study of Sodium Xylenesulfonate

Mean Body Weightb (g) Final Weight a Dose Survival Initial Final Change Relative to Controls

(mg/mL) (%) 

Male

0 5/5 23.7 ± 0.9 25.4 ± 0.8 1.7 ± 0.3 

5 5/5 24.0 ± 0.5 24.8 ± 1.8 0.8 ± 1.6 98 

15 5/5 23.2 ± 0.6 26.1 ± 0.7 2.9 ± 0.2 103 

44 5/5 22.3 ± 0.3 25.0 ± 0.3 2.6 ± 0.2 98 

133 5/5 23.7 ± 0.4 25.6 ± 0.4 1.9 ± 0.3 101 

400 5/5 23.3 ± 0.4 26.4 ± 0.3 3.1 ± 0.2 104 

Female

0 5/5 18.4 ± 0.2 20.9 ± 0.4 2.5 ± 0.3 

5 5/5 17.9 ± 0.5 20.2 ± 0.3 2.4 ± 0.4 97 

15 5/5 18.2 ± 0.3 20.8 ± 0.5 2.6 ± 0.4 100 

44 5/5 18.8 ± 0.5 20.8 ± 0.5 2.0 ± 0.3 99 

133 5/5 18.2 ± 0.3 21.0 ± 0.3 2.8 ± 0.4 100 

400 5/5 18.1 ± 0.4 21.3 ± 0.2 3.2 ± 0.4 102 

a Number of animals surviving at 17 days/number initially in group b Weights and weight changes are given as mean ± standard error. Differences from the control group were not significant by Dunnett’s test. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

36 Sodium Xylenesulfonate, NTP TR 464 

14-WEEK STUDY

There were no chemical-related deaths (Table 10). 

The mean body weight gain of 400 mg/mL males

was significantly greater than that of the control

group. Dermal applications of 100 μL of 5, 15, 44, 

133, and 400 mg/mL delivered average daily doses of

approximately 17, 40, 140, 440, and 1,300 mg

sodium xylenesulfonate/kg body weight to males and

20, 60, 170, 530, and 1,630 mg/kg to females. There

were no clinical findings related to sodium

xylenesulfonate administration. 

Epidermal hyperplasia occurred in 44 mg/mL

females, 133 mg/mL males, and 400 mg/mL males

and females (Table 11). Hyperplasia of the epidermis

in 400 mg/mL males and females was considered

related to chemical administration. Epidermal hyperplasia was most pronounced in male mice where it

typically developed as a mild multifocal nodular to

papillary thickening of the epidermis. The involved

epidermis was somewhat disorganized, and cells

tended to be elongated (oval or columnar) rather than

normal cuboidal epidermal epithelium. 

Chronic inflammation of the skin occurred primarily

in three control and one dosed animal of each gender. 

These lesions consisted of mononuclear inflammatory

cells in the dermis. Chronic inflammation was

frequently accompanied by a minimal epidermal

hyperplasia which was clearly a reaction to the

inflammation and not diagnosed separately. 

TABLE 10 

Survival and Body Weights of Mice in the 14-Week Dermal Study of Sodium Xylenesulfonate

Mean Body Weightb (g)	# Final Weight

Dose

(mg/mL) 

Survivala Initial Final Change Relative to Controls

(%) 

Male

0 

5 

15 

44 

133 

400 

10/10 

10/10 

10/10 

10/10 

10/10 

10/10 

25.9 ± 0.5 

25.5 ± 0.5 

25.9 ± 0.4 

25.9 ± 0.5 

25.2 ± 0.4 

25.5 ± 0.6 

35.7 ± 0.5 

34.5 ± 0.6 

35.5 ± 0.7 

36.6 ± 0.7 

35.7 ± 0.7 

37.4 ± 0.7 

9.8 ± 0.5 

9.0 ± 0.5 

9.6 ± 0.4 

10.7 ± 0.4 

10.5 ± 0.6 

11.9 ± 1.0* 

96 

100 

102 

100 

105 

Female

0 

5 

15 

44 

133 

400 

10/10 

9/10 c

9/10c

10/10 

9/10c

10/10 

19.0 ± 0.3 

19.0 ± 0.3 

19.1 ± 0.3 

19.5 ± 0.2 

19.4 ± 0.4 

19.1 ± 0.3 

30.8 ± 0.7 

29.6 ± 0.6 

31.0 ± 0.8 

32.6 ± 0.5 

30.4 ± 0.6 

30.0 ± 0.5 

11.9 ± 0.6 

10.6 ± 0.6 

12.2 ± 0.7 

13.2 ± 0.6 

11.4 ± 0.6 

10.9 ± 0.4 

96 

101 

106 

99 

97 

*	#Significantly different (P#0.05) from the control group by Williams’ test a Number of animals surviving at 14 weeks/number initially in group b	 Weights and weight changes are given as mean ± standard error. Subsequent calculations are based on animals surviving to the end of the

study. 

One mouse was removed from study on day 8 due to pregnancy. c
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

37 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE 11 

Incidences of Epidermal Hyperplasia in Mice in the 14-Week Dermal Study

of Sodium Xylenesulfonate

Vehicle 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

Control

Male

Skin, Site of Applicationa

Epidermal Hyperplasiab

10 

0 

10 

0 

10 

0 

10 

0 

10 

2 (1.0) c 10 5* (1.8) 

Female

Skin, Site of Application

Epidermal Hyperplasia

10 

0 

9 

0 

9 

0 

10 

1 (1.0) 

9 

0

10 

4* (1.0) 

* Significantly different (P#0.05) from the vehicle control by the Fisher exact test a Number of mice with skin examined microscopically b Number of mice with lesion
 

Average severity grade of lesion in affected animals: 1  ̆ minimal; 2  ̆ mild; 3  ̆ moderate; 4  ̆ marked
 

Dose Selection Rationale: In the 17-day and 14-week deliver the 727 mg/kg dose) in the 2-year study. 

studies, dose concentrations of up to 400 mg/mL were Fixed volumes and concentrations were used in the

well tolerated. The only limitation to the concen- 17-day and 14-week studies which resulted in

tration tration used in dermal dosing in the 2-year decreasing dose concentrations (mg/kg body weight) 

study was the difficulty of obtaining a uniform as the animals grew (Appendix I). Constant dose

suspension of sodium xylenesulfonate in the vehicle. concentrations were achieved in the 2-year study by

To obtain uniform suspensions, the highest dose adjusting the applied dose volumes to the body

concentration was decreased to 300 mg/mL (used to weights throughout the study. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

38 Sodium Xylenesulfonate, NTP TR 464 

2-YEAR STUDY

Survival

Estimates of 2-year survival probabilities for male and

female mice are shown in Table 12 and in the KaplanMeier survival curves (Figure 3). Survival of dosed

males and females was similar to that of the control

groups. 

TABLE 12 

Body Weights and Clinical Findings

Mean body weights of dosed males and females were

generally similar to those of the controls throughout

the study; however, the mean body weight of

727 mg/kg females was greater than that of the

control group from week 85 to week 97 (Tables 13 

and 14, Figure 4). With the exception of irritation at

the site of application in one 364 mg/kg female, there

were no clinical findings related to sodium xylenesulfonate administration. 

Survival of Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Male

Animals initially in study	 50 50 50 50 

Moribund 7 3 8 6 

Natural deaths 11 10 3 9 

Animals surviving to study termination 32 37 39 35 a Percent probability of survival at end of study 64 74 78 70 

Mean survival (days)

b 704 704 710 705 

c Survival analysis	 P ̆0.681N P ̆0.440N P ̆0.218N P ̆0.635N

Female

Animals initially in study	 50 50 50 50 

Missingd 0 1 0 0 

Moribund 12 7 9 6 

Natural deaths 7 10 9 8 

Animals surviving to study termination 31 32 32 36 

Percent probability of survival at end of study 62 66 64 72 

Mean survival (days) 690 679 704 703 

Survival analysis	 P ̆0.305N P ̆0.896N P ̆0.967N P ̆0.335N

a Kaplan-Meier determinations

b	 Mean of all deaths (uncensored, censored, and terminal sacrifice) 

The result of the life table trend test (Tarone, 1975) is in the vehicle control column, and the results of the life table pairwise comparisons

(Cox, 1972) with the vehicle controls are in the dosed columns. A negative trend or lower mortality in a dose group is indicated by N. d	 Censored from survival analyses

c
 

Sodium Xylenesulfonate, NTP TR 464 39 

FIGURE 3
 

Kaplan-Meier Survival Curves for Male and Female Mice
 

Administered Sodium Xylenesulfonate Dermally for 2 Years
 
 

 

40 Sodium Xylenesulfonate, NTP TR 464 

TABLE 13 

Mean Body Weights and Survival of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

Weeks Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

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 23.5 50 23.5 100 50 23.8 101 50 23.6 100 50 

2 23.7 50 24.3 103 50 24.2 102 50 24.5 103 50 

3 25.8 50 26.0 101 50 25.9 100 50 26.4 102 50 

4 26.0 50 26.3 101 50 26.5 102 50 26.6 102 50 

5 27.0 50 27.0 100 50 27.5 102 50 27.4 102 50 

6 28.4 50 28.5 100 50 28.6 101 50 28.7 101 50 

7 28.3 50 28.4 100 50 28.6 101 50 28.9 102 50 

8 29.0 50 29.3 101 50 29.3 101 50 29.4 101 50 

9 30.2 50 30.4 101 50 30.4 101 50 30.6 101 50 

10 30.2 50 30.7 102 50 30.3 100 50 31.1 103 50 

11 31.2 50 31.9 102 50 31.6 101 50 32.1 103 50 

12 32.3 50 32.6 101 50 32.2 100 50 32.5 101 50 

13 33.3 50 33.7 101 50 33.2 100 50 33.8 102 50 

17 35.7 50 36.1 101 50 35.8 100 50 36.2 101 50 

21 37.7 50 38.3 102 50 37.5 100 50 38.6 102 50 

25 38.6 50 39.5 102 50 38.9 101 50 40.1 104 50 

28 41.3 50 41.8 101 50 41.0 99 50 42.5 103 50 

33 41.9 50 42.1 101 50 41.6 99 50 42.3 101 50 

37 43.2 50 43.9 102 50 43.1 100 50 43.6 101 50 

41 43.4 50 44.1 102 50 43.4 100 50 43.9 101 50 

45 44.8 50 44.9 100 50 44.5 99 50 45.3 101 50 

49 45.1 50 45.8 102 49 44.6 99 50 46.0 102 50 

53 46.1 50 46.0 100 49 45.3 98 50 46.5 101 50 

57 45.6 50 46.9 103 49 45.9 101 50 47.1 103 50 

61 46.6 50 46.9 101 49 46.5 100 50 46.9 101 50 

65 46.5 50 47.1 101 49 46.7 100 50 46.8 101 50 

69 47.2 50 47.1 100 49 46.7 99 50 47.6 101 49 

73 46.9 49 47.0 100 49 46.4 99 50 47.5 101 49 

77 46.8 49 46.9 100 49 46.6 100 49 47.8 102 49 

80 45.8 49 46.2 101 48 46.1 101 48 47.1 103 48 

85 46.0 46 45.3 99 47 46.1 100 46 46.9 102 46 

89 44.7 46 44.3 99 46 45.6 102 45 46.1 103 46 

93 42.8 46 42.9 100 44 44.4 104 43 44.3 104 43 

97 42.6 43 42.6 100 42 43.8 103 43 44.3 104 41 

101 42.5 35 42.0 99 39 43.6 103 41 43.2 102 40 

105 41.5 32 40.0 96 37 42.0 101 39 42.2 102 35 

Mean for weeks

1-13 28.4 28.7 101 28.6 101 28.9 102 

14-52 41.3 41.8 101 41.2 100 42.1 102 

53-105 45.1 45.1 100 45.4 101 46.0 102 
 

 

Sodium Xylenesulfonate, NTP TR 464 41 

TABLE 14 

Mean Body Weights and Survival of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

Weeks Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

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 18.7 50 18.9 101 50 18.8 101 50 18.9 101 50 

2 19.6 50 20.4 104 50 20.0 102 50 20.1 103 50 

3 22.0 50 22.2 101 50 22.3 101 50 22.0 100 50 

4 22.4 50 22.6 101 50 22.5 100 50 22.7 101 50 

5 22.9 50 23.5 103 49 23.4 102 50 23.5 103 50 

6 24.1 50 24.9 103 49 24.9 103 50 25.2 105 50 

7 24.7 50 25.0 101 49 25.1 102 50 25.1 102 50 

8 25.3 50 25.6 101 49 25.7 102 50 25.7 102 50 

9 26.2 50 26.6 102 49 26.5 101 50 26.5 101 50 

10 26.9 50 27.3 102 49 27.1 101 50 27.2 101 50 

11 27.0 50 27.5 102 49 27.7 103 50 27.5 102 50 

12 27.9 50 28.2 101 49 27.7 99 50 28.0 100 49 

13 28.8 50 29.6 103 49 29.2 101 50 29.1 101 49 

17 30.6 50 31.3 102 49 30.8 101 50 30.6 100 49 

21 33.0 50 33.7 102 49 32.8 99 50 32.9 100 49 

25 34.0 50 35.4 104 49 34.2 101 50 34.1 100 49 

28 37.3 50 38.2 102 49 37.4 100 50 36.9 99 49 

33 38.0 50 38.8 102 49 37.9 100 50 37.4 98 49 

37 40.2 50 40.9 102 49 40.0 100 50 39.6 99 49 

41 41.0 50 41.6 102 49 40.6 99 50 40.8 100 49 

45 42.4 49 43.2 102 49 41.9 99 50 42.3 100 49 

49 43.0 49 44.4 103 48 42.7 99 50 42.9 100 49 

53 45.0 49 45.6 101 48 44.1 98 50 44.7 99 49 

57 45.0 49 46.9 104 48 45.0 100 50 46.2 103 49 

61 46.2 49 47.4 103 47 46.1 100 50 47.2 102 48 

65 47.6 49 48.4 102 46 46.8 98 50 48.8 103 48 

69 49.0 47 50.1 102 45 48.1 98 50 50.1 102 48 

73 49.1 45 50.8 104 45 49.1 100 50 51.2 104 48 

77 50.1 45 51.6 103 45 49.3 98 50 51.7 103 48 

81 50.6 45 52.3 103 44 49.3 97 50 52.7 104 47 

85 50.1 44 52.7 105 44 50.3 100 46 53.0 106 47 

89 48.8 44 51.0 105 43 49.8 102 44 51.9 106 46 

93 47.9 41 48.8 102 39 49.0 102 41 51.3 107 44 

97 47.3 38 49.4 104 37 49.0 104 37 50.9 108 44 

101 47.2 34 47.5 101 36 48.9 104 34 49.2 104 40 

105 45.8 31 44.9 98 33 46.5 102 32 46.6 102 37 

Mean for weeks

1-13 24.3 24.8 102 24.7 102 24.7 102 

14-52 37.7 38.6 102 37.6 100 37.5 99 

53-105 47.8 49.1 103 48.0 100 49.7 104 
 

42 Sodium Xylenesulfonate, NTP TR 464 

FIGURE 4
 

Growth Curves for Male and Female Mice Administered
 

Sodium Xylenesulfonate Dermally for 2 Years
 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 43 

Pathology and Statistical Analysis

This section describes the statistically significant and

biologically noteworthy changes in the incidences of

neoplasms and nonneoplastic lesions of the skin and

liver. Summaries of the incidences of neoplasms and

nonneoplastic lesions, individual animal tumor

diagnoses and statistical analyses of primary neoplasms that occurred with incidences of at least 5% in

at least one animal group, and historical incidences for

the neoplasms mentioned in this section are presented

in Appendix C for male mice and Appendix D for

female mice. 

Skin: The incidences and morphologies of the

neoplasms that occurred in males and females were

typical of spontaneous neoplasms in B6C3F1 mice

and were not considered related to treatment

(Tables 15, C1, and D1). 

Hyperplasia of the epidermis occurred in control and

364 and 727 mg/kg males and in control and dosed

females (Tables 15, C5, and D5). These lesions

consisted of minimal to mild multifocally extensive

thickening of the layers of epidermal epithelium. In

males, there was a possible treatment-related effect in

the incidences of epidermal hyperplasia as the

incidences occurred with a positive (P#0.05) trend; 

however, other nonneoplastic lesions of the skin at the

site of application were not considered related to

chemical administration. Focal ulceration occurred in

one 727 mg/kg male and in one female in each dose

group (Tables 15 and D5). Typically, sites of

ulceration had superficial accumulations of protein

and necrotic cell debris, and the exposed dermis had

infiltrates of both neutrophils and mononuclear

inflammatory cells. Exudates of cell debris and

protein on the surface of the epidermis without

evidence of ulceration were diagnosed separately, as

was inflammation in the dermis or subcutaneous

tissues. 

TABLE 15 

Incidences of Neoplasms and Nonneoplastic Lesions of the Skin of Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Male

Skin, Site of Applicationa

Epidermis, Exudateb

Epidermis, Hyperkeratosis

Epidermis, Hyperplasia

Epidermis, Ulcer

50 

0 

0 

1 

0 

(1.0) 

50 

1 

0 

0 

0 

(1.0)c 50 

1 

1 

4 

0 

(1.0) 

(2.0) 

(1.3) 

50 

1 

0 

5 

1 

(3.0) 

(1.4) 

(3.0) 

Subcutaneous Tissue, Hemangioma

Subcutaneous Tissue, Hemangiosarcoma

Subcutaneous Tissue, Histiocytic Sarcoma

Subcutaneous Tissue, Malignant Lymphoma

0 

0 

0 

0 

0 

0 

2 

0 

1 

1 

0 

1 

0 

0 

0 

0 

Female

Skin, Site of Application

Epidermis, Exudate

Epidermis, Hyperplasia

Epidermis, Ulcer

Inflammation, Chronic

Parakeratosis

50 

1 

4 

0 

4 

0 

(2.0) 

(1.3) 

(1.0) 

49 

4 

1 

1 

1 

0 

(1.0) 

(2.0) 

(2.0) 

(2.0) 

50 

4 

4 

1 

1 

1 

(1.5)

(1.5) 

(2.0) 

(1.0) 

(1.0) 

50 

1 

4 

1 

1 

0 

(4.0) 

(1.5) 

(3.0) 

(1.0) 

Subcutaneous Tissue, Fibrosarcoma 0 1 1 0 

a

b

c

Number of mice with skin examined microscopically

Number of mice with lesion

Average severity grade of lesions in affected animals: 1  ̆ minimal; 2  ̆ mild; 3  ̆ moderate; 4  ̆ marked
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

44 Sodium Xylenesulfonate, NTP TR 464 

Inflammation in the skin of female mice was consisted of mononuclear cells and sometimes lesser

subclassified according to the inflammatory cells numbers of neutrophils centered around bile ducts but

present: suppurative (composed of neutrophils); also focally in the parenchyma. Primarily in male

chronic active (neutrophils and inflammatory cells); mice, this diagnosis also included various degrees of

and chronic (mononuclear inflammatory cells). hepatocytomegaly, hepatocyte degeneration, fibrosis

Hyperkeratosis occurred in a single 364 mg/kg male and/or regenerative hyperplasia of hepatocytes. Bile

and consisted of focally thickened layers of keratin duct hyperplasia included oval (bile ductule) cell

overlying the epidermis (Tables 15 and C5). proliferation as well as increased numbers of bile

Parakeratosis in a single 364 mg/kg female was a ducts. Chronic active inflammation and bile duct

superficial accumulation of desquamated epidermal hyperplasia, typically graded as mild to moderate, 

cells that retained obvious nuclei (Tables 15 and D5). were both noted in more than 80% of control and

dosed male mice (Table 16). In female mice, 

Liver: In males and females from control and dosed reviewing pathologists identified minimal severity

groups, incidences of hepatocellular adenoma, chronic active inflammation in approximately half of

hepatocellular carcinoma, hepatocellular adenoma or both control and dosed groups, whereas bile duct

carcinoma (combined), chronic active inflammation hyperplasia, typically minimal, was identified in only

(hepatitis), and hyperplasia of bile ductular epithelium two or four animals in each group. Steiner’s

(males only) were generally greater than those modification of the Warthin-Starry stain was applied

expected by spontaneous occurrence (Tables 16, C1, to the livers of 10 male and 10 female mice with

C5, D1, and D5). The hepatocellular neoplasm lesions of chronic-active inflammation and bile duct

incidences exceeded those in historical controls from hyperplasia. Organisms consistent with Helicobacter

2-year NTP dermal and feed studies (Tables 16, C4, hepaticus bacteria were identified in the livers of six

and D4). Chronic active inflammation in the liver males, but in none of the females (Appendix L). 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

45 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE 16 

Incidences of Neoplasms and Nonneoplastic Lesions of the Liver in Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Male

Number Examined Microscopically

Inflammation, Chronic Activea 50 46 (2.5)b

50 

44 (2.8) 

50 

41 (2.5) 

50 

43 (2.4) 

Bile Duct Hyperplasia 43 (2.5) 41 (2.8) 36 (2.6) 37 (2.6) 

Hepatocellular Adenoma (includes multiple) 37 32 21** 29 

Hepatocellular Carcinoma (includes multiple) 35 31 22** 26 

Hepatocellular Adenoma or Carcinomac 46 41 31 45 

Hepatoblastoma 0 4 0 2 

Female

Number Examined Microscopically 50 49 50 50 

Inflammation, Chronic Active 22 (1.0) 28 (1.1) 24 (1.1) 21 (1.1) 

Bile Duct Hyperplasia 4 (1.8) 2 (1.0) 4 (1.0) 2 (1.5) 

Hepatocellular Adenoma (includes multiple) 18 17 18 28 

Hepatocellular Carcinoma (includes multiple) 

Hepatocellular Adenoma or Carcinomad

10 

27 

13 

23 

7 

23 

10 

33 

** Significantly different (P#0.01) from the vehicle control by the logistic regression test a Number of mice with lesion

b Average severity grade of lesion in affected animals: 1  ̆ minimal; 2  ̆ mild; 3  ̆ moderate; 4  ̆ marked

Historical incidence for 2-year NTP dermal (acetone or ethanol vehicle) or feed studies with undosed control groups (mean ± standard

deviation): acetone: 63/150 (42.0% ± 22.3%); range, 18%-62%; ethanol: 29/50 (58.0%); feed: 596/1,465 (40.7% ± 14.5%); range, 

10%-68% 

d Historical incidence: acetone: 40/150 (26.7% ± 17.0%); range, 14%-46%; ethanol: 27/52 (51.9%); feed: 313/1,464 (21.4% ± 13.0%); 

range, 3%-56% 

G Without S9, no significant increase in mutations was ENETIC TOXICOLOGY

Sodium noted. Sodium xylenesulfonate induced dose-related xylenesulfonate (100 to 10,000 μg/plate) was

not mutagenic in Salmonella typhimurium strain increases in sister chromatid exchanges in cultured

TA98, TA100, TA1535, or TA1537, with or without Chinese hamster ovary cells at concentrations that

induced S9 (Table E1). Results obtained with sodium produced cell cycle delay (2,513 to 5,000 μg/mL) in

xylenesulfonate in a mammalian gene mutation assay the absence of S9; with S9, no increases in sister

with cultured L5178Y mouse lymphoma cells in the chromatid exchanges were noted (Table E3). Finally, 

presence of S9 (Table E2) were concluded to be no induction of chromosomal aberrations was

equivocal because the significant increase in mutant observed in cultured Chinese hamster ovary cells

colonies noted in the first trial with S9 was not treated with sodium xylenesulfonate (2,513 to

convincingly demonstrated in the second trial. 5,000 μg/mL) with or without S9 (Table E4). 
46 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

47 

DISCUSSION AND CONCLUSIONS
 

Sodium xylenesulfonate is used as a hydrotrope in sodium xylenesulfonate is less in ethanol than in

liquid household detergents and in shampoos; thus, water, the 400 mg/mL concentration was found to

dermal exposure to human skin is widespread. require constant agitation during dosing in order to

Consumers may be exposed to solutions containing up avoid settling of the chemical. Therefore, to obtain

to 10% (100 mg/mL) sodium xylenesulfonate more uniform suspensions, the highest concentration

(Phil Tham, Reckitt and Colman, Inc., personal used in the 2-year studies was 300 mg/mL. Constant

communication, 1995). In 1992, United States volumes were applied during the 17-day and 14-week

production of sodium xylenesulfonate was approx- studies, and because the animals gained weight during

imately 30,000 tons (USITC, 1994). The present the studies, the dose concentrations (mg/kg body

studies are the first to address the possible dermal weight) decreased as the animals grew (Appendix I). 

toxicity and carcinogenicity of sodium To maintain constant dose concentrations throughout

xylenesulfonate. the 2-year studies, dose volumes were adjusted

according to body weight as the animals aged. These

Technical grade sodium xylenesulfonate contains modifications resulted in less chemical treatment in

approximately 35% sodium ethylbenzenesulfonate and the 2-year studies than in the shorter duration studies. 

11.5% ortho-, 38% meta-, and 15.5% para-sodium The actual amount of chemical treatment in the

xylenesulfonate (Mausner and Sosis, 1962). Although shorter duration studies, however, was probably less

sodium xylenesulfonate is sometimes classified with than the calculated amount because of the problems of

surface-active agents, its alkyl chain is too short to rapid removal in the 17-day studies and of nonconfer surface active properties (Kirk-Othmer, 1994). uniform suspensions in the 14-week studies. 

As a hydrotrope, sodium xylenesulfonate increases the

solvent capacity of water toward other molecules, Use of 50% ethanol as a vehicle rather than water

unlike surfactants that solubilize by enclosing solutes increased the probability of skin penetration

in micelles (Hunting, 1983). Because of the polar (Grandjean, 1988). However, the low dermal toxicity

sulfonate group, dermal absorption of sodium xylene- of sodium xylenesulfonate in 50% ethanol in the

sulfonate is probably poor (Grandjean et al., 1988). present studies does not rule out dermal toxicity of

Because ethanol has a higher permeability constant for more complex mixtures which might further affect

the skin than water (Scheuplein and Blank, 1971), the penetration or reactivity of the skin, nor do the

use of a 50% ethanol vehicle in the 14-week and present studies address the toxicity of complex

2-year studies may have enhanced absorption over mixtures containing sodium xylenesulfonate by other

that in the 17-day studies, in which the vehicle was routes of exposure, such as occupational inhalation of

water. As in all dermal studies, there was potential aerosolized compound. 

for some ingestion as animals groomed themselves. 

Dermal application of 400 mg/mL sodium xyleneModifications were made to the dosing regimens sulfonate did not significantly affect survival or mean

between the different duration studies. Water was body weights of rats or mice dosed for 17 days or

used as a vehicle for sodium xylenesulfonate for the 14 weeks. Similarly, dermal applications of

17-day studies, but this vehicle was changed to 50% 300 mg/mL sodium xylenesulfonate did not signifiethanol for the longer duration studies because the cantly affect the survival or mean body weights of rats

suspensions in water were noted to bead up on the and mice dosed for 2 years. Survival of male rats to

skin and were easily shaken off by the animals. Dose termination of the 2-year study was 18% in the

concentrations of up to 400 mg/mL technical grade control and 120 mg/kg groups and 36% and 26% in

sodium xylenesulfonate in 50% alcohol were applied the 60 and 240 mg/kg groups. Survival was low in

in the 14-week studies, but because the solubility of comparison to studies in the NTP database in which
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

48 Sodium Xylenesulfonate, NTP TR 464 

rats were not housed individually, but survival was

consistent with other studies utilizing individual

housing (Rao, 1995). It has been postulated that the

social interaction afforded by multiple housing

enhances the well-being of rats (Rao, 1995), whereas

individual housing is detrimental to survival, particularly of males. 

In rats in the 17-day and 14-week studies, the only

treatment-related clinical finding was brownish

discoloration at the site of application. The sebaceous

gland hyperplasia noted in both control and dosed rats

in the 2-year study suggests that fur-clipping and

vehicle application may stimulate sebaceous glands

and the production of sebum in this species. The

brownish discoloration of the skin may have resulted

from the combinations of sebum and sodium

xylenesulfonate. 

There was some indication that the incidences, but not

the severities, of epidermal hyperplasia were related

to chemical administration in male and female rats and

mice in the 14-week studies and in female rats and

male mice in the 2-year studies. In the 14-week

studies, epidermal hyperplasia occurred in control and

dosed groups and was less apparent in rats than in

mice. In the 2-year study, rats with epidermal

hyperplasia had slightly more extensive and thicker

lesions than in the 14-week study, but the lesions were

typically minimal in severity. 

Some male mice in the 14-week study had papillary

thickening of the epidermis. Based on results of the

2-year study, there was no evidence that these lesions

would have progressed to skin neoplasms in time. 

Further, there was less focally exuberant epidermal

hyperplasia in male mice in the 2-year study than in

male mice in the 14-week study. Because epidermal

hyperplasia occurred in control groups as well as

dosed groups in the 14-week study, these lesions may

have been due to irritation from weekly fur clipping

and vehicle application. Comparatively lower incidences of hyperplasia in the 2-year study may reflect

physiologic adaptation to treatment, resolution of

lesions during the 10 days between the last dose and

necropsy, or the lower dosing concentration used in

the 2-year study. 

Ten percent or less of the rats and mice in the 2-year

studies developed epidermal hyperplasia. These

lesions were typically minimal to mild in severity. In

male and female rats and in male mice, there was a

suggestion that sodium xylenesulfonate exacerbated

epidermal hyperplasia; however, the incidences in

dosed groups did not differ significantly from each

other or from those in the control groups. In female

mice, the incidences and severities of epidermal

hyperplasia in dosed and control groups of mice were

similar. In addition to epidermal hyperplasia, 

pinpoint ulcers of the epidermis occurred in up to two

animals per group in dosed female rats and dosed

male and female mice and in one control female rat. 

The low incidences and severities of these lesions lead

to the conclusion that sodium xylenesulfonate is not

dermally toxic. Most lesions were also observed in

control animals receiving only the vehicle, which

suggests that the occurrence of these lesions was

related to weekly fur clipping and vehicle application

rather than to the application of sodium xylenesulfonate. In both rats and mice, neoplasms that

occurred in the skin at the site of application in the

2-year studies were typical in morphology and

incidence to spontaneously occurring lesions in

F344/N rats and B6C3F1 mice. None were considered related to chemical administration. 

The only neoplasm showing a possible chemicalrelated effect in the 2-year study was mononuclear cell

leukemia, of which the incidences were marginally

increased in the 60 and 240 mg/kg male rat groups. 

The lack of statistical significance by the life table

analysis (an appropriate test for this generally fatal

neoplasm) and the lack of a clear dose-response

relationship suggest that these increases were

unrelated to chemical administration. 

Incidences of hepatocellular carcinoma and hepatocellular adenoma and carcinoma (combined) in all

groups of male mice exceeded the NTP historical

control ranges. Female mice in the control and

727 mg/kg groups had incidences of hepatocellular

carcinoma that matched the highest control incidence, 

and the combined incidence of hepatocellular adenoma

and carcinoma in 727 mg/kg females exceeded the

historical control range. Chronic active inflammation, 

bile duct hyperplasia, and organisms consistent with

the recently discovered bacteria Helicobacter

hepaticus were observed in the livers of male mice in

these studies. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

__________

 

 

Sodium Xylenesulfonate, NTP TR 464 49 

Based on retrospective analyses, H. hepaticus was

determined to have infected mice in 12 recent NTP

2-year studies (Appendix L). Of the 12 studies, mice

(primarily males) from nine studies (including this

study of sodium xylenesulfonate) had H. hepaticusassociated hepatitis. Qualitatively, the hepatitis and

silver-staining organisms within the liver were similar

among the nine studies. With assays based on

polymerase chain reaction, H. hepaticus was

identified in studies from which adequately preserved

(frozen) liver tissue was available. In general, efforts

to identify H. hepaticus from tissue fixed in formalin

for over a week were not successful (Malarkey et al., 

1997), which was the case for this study of sodium

xylenesulfonate. However, because of the presence of

the typical liver lesions and silver-positive helical

organisms, mice from this study were presumed to be

infected with H. hepaticus. 

Increases in the incidence of hepatocellular neoplasms

in male mice have been shown to be associated with

H. hepaticus infection when hepatitis is also present

(Ward et al., 1994; Fox et al., 1996; Appendix L). 

Additionally, within NTP studies with H. hepaticusassociated hepatitis, increased incidences of

hemangiosarcoma were seen in the livers of male mice

(Appendix L.) 

Because of these associations, interpretation of a

treatment-related increase, but not a lack of response, 

in the incidences of these two neoplasm types in the

liver of male mice is considered confounded. 

Chemical-related responses (or lack thereof) at other

sites in male mice and in female mice are not

considered to have been significantly impacted by the

infection with H. hepaticus or its associated hepatitis

(Appendix L). In the present studies of sodium

xylenesulfonate, the incidences of hepatocellular

neoplasms were not increased with dose; thus it is

concluded that the presence of H. hepaticus did not

obscure any potential chemical-related neoplasia. 

CONCLUSIONS

Under the conditions of these 2-year dermal studies, 

there was no evidence of carcinogenic activity* of

sodium xylenesulfonate in male or female F344/N rats

administered 60, 120, or 240 mg/kg or in male or

female B6C3F1 mice administered 182, 364, or

727 mg/kg. 

Increased incidences of epidermal hyperplasia in

female rats and male mice may have been related to

exposure to sodium xylenesulfonate. 

* 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. 
50 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

51 

REFERENCES
 

Armitage, P. (1971). Statistical Methods in Medical

Research, pp. 362-365. John Wiley and Sons, Inc., 

New York. 

Ashby, J., and Tennant, R.W. (1991). Definitive

relationships among chemical structure, carcinogenicity and mutagenicity for 301 chemicals tested by

the U.S. NTP. Mutat. Res. 257, 229-306. 

Boorman, G.A., Montgomery, C.A., Jr., 

Eustis, S.L., Wolfe, M.J., McConnell, E.E., and

Hardisty, J.F. (1985). Quality assurance in pathology

for rodent carcinogenicity studies. In Handbook of

Carcinogen Testing (H.A. Milman and E.K. 

Weisburger, Eds.), pp. 345-357. Noyes Publications, 

Park Ridge, NJ. 

Caspary, W.J., Lee, Y.J., Poulton, S., Myhr, B.C., 

Mitchell, A.D., and Rudd, C.J. (1988). Evaluation of

the L5178Y mouse lymphoma cell mutagenesis assay: 

Quality-control guidelines and response categories. 

Environ. Mol. Mutagen. 12 (Suppl. 13), 19-36. 

Code of Federal Regulations (CFR) 21, Part 58. 

Cox, D.R. (1972). Regression models and life-tables. 

J. R. Stat. Soc. B34, 187-220. 

Crawford, B.D. (1985). Perspectives on the somatic

mutation model of carcinogenesis. In Advances in

Modern Environmental Toxicology. Mechanisms and

Toxicity of Chemical Carcinogens and Mutagens

(M.A. Mehlman, W.G. Flamm, and R.J. Lorentzen, 

Eds.), pp. 13-59. Princeton Scientific Publishing

Co., Princeton, NJ. 

Dinse, G.E., and Haseman, J.K. (1986). Logistic

regression analysis of incidental-tumor data from

animal carcinogenicity experiments. Fundam. Appl. 

Toxicol. 6, 44-52. 

Dinse, G.E., and Lagakos, S.W. (1983). Regression

analysis of tumour prevalence data. Appl. Statist. 32, 

236-248. 

Dixon, W.J., and Massey, F.J., Jr. (1951). Introduction to Statistical Analysis, 1st ed., pp. 145-147. 

McGraw-Hill Book Company, Inc., New York. 

Dunn, O.J. (1964). Multiple comparisons using rank

sums. Technometrics 6, 241-252. 

Dunnett, C.W. (1955). A multiple comparison

procedure for comparing several treatments with a

control. J. Am. Stat. Assoc. 50, 1096-1121. 

Fox, J.G., MacGregor, J., Shen Z., Li, X., 

Lewis, R., and Dangler, C.A. (1998). Role of

Helicobacter hepaticus in confounding results of a

triethanolamine carcinogenesis study in mice (in

preparation). 

Galloway, S.M., Armstrong, M.J., Reuben, C., 

Colman, S., Brown, B., Cannon, C., Bloom, A.D., 

Nakamura, F., Ahmed, M., Duk, S., Rimpo, J., 

Margolin, B.H., Resnick, M.A., Anderson, B., and

Zeiger, E. (1987). Chromosome aberrations and

sister chromatid exchanges in Chinese hamster ovary

cells: Evaluations of 108 chemicals. Environ. Mol. 

Mutagen. 10 (Suppl. 10), 1-175. 

Gart, J.J., Chu, K.C., and Tarone, R.E. (1979). 

Statistical issues in interpretation of chronic bioassay

tests for carcinogenicity. J. Natl. Cancer Inst. 62, 

957-974. 

Grandjean, P., Berlin, A., Gilbert, M., and

Penning, W. (1988). Preventing percutaneous

absorption of industrial chemicals: The “skin”

denotation. Am. J. Ind. Med. 14, 97-107. 

Haseman, J.K. (1984). Statistical issues in the

design, analysis and interpretation of animal carcinogenicity studies. Environ. Health Perspect. 58, 

385-392. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

52 Sodium Xylenesulfonate, NTP TR 464 

Hollander, M., and Wolfe, D.A. (1973). Nonparametric Statistical Methods, pp. 120-123. John Wiley

and Sons, New York. 

Hunting, A.L.L. (1983). Encyclopedia of Shampoo

Ingredients, Micelle Press, Cranford, NJ. 

Jonckheere, A.R. (1954). A distribution-free

k-sample test against ordered alternatives. Biometrika

41, 133-145. 

Kaplan, E.L., and Meier, P. (1958). Nonparametric

estimation from incomplete observations. J. Am. Stat. 

Assoc. 53, 457-481. 

Kirk-Othmer Encyclopedia of Chemical Technology

(1994) 3rd ed., Vol. 22, p. 351. John Wiley and

Sons, Inc., New York. 

Kondratiuk, V.A., Pastushenko, T.V., Sergeta, V.N., 

Gun’ko, L.M., Pereima, V.I., Golka, N.V., 

Gnatiuk, M.S., and Kalashnik, S.A. (1983). 

Hygienic regulation of sodium toluene sulfonate in

reservoir water. Gig. Sanit. 8, 67-69. 

McConnell, E.E., Solleveld, H.A., Swenberg, J.A., 

and Boorman, G.A. (1986). Guidelines for

combining neoplasms for evaluation of rodent

carcinogenesis studies. JNCI 76, 283-289. 

McKnight, B., and Crowley, J. (1984). Tests for

differences in tumor incidence based on animal

carcinogenesis experiments. J. Am. Stat. Assoc. 79, 

639-648. 

Malarkey, D.E., Ton, T.-V., Hailey, J.R., and

Devereaux, T.R. (1997). A PCR-RFLP method for

the detection of Helicobacter hepaticus in frozen or

fixed liver from B6C3F1 mice. Toxicol. Pathol. 25, 

606-612. 

Maronpot, R.R., and Boorman, G.A. (1982). 

Interpretation of rodent hepatocellular proliferative

alterations and hepatocellular tumors in chemical

safety assessment. Toxicol. Pathol. 10, 71-80. 

Mausner, M., and Sosis, P. (1962). Hydrotropes in

liquid syndets. Soaps Chem. Spec. 38, 47. 

Miller, J.A., and Miller, E.C. (1977). Ultimate

chemical carcinogens as reactive mutagenic electrophiles. In Origins of Human Cancer (H.H. Hiatt, 

J.D. Watson, and J.A. Winsten, Eds.), pp. 605-627. 

Cold Spring Harbor Laboratory, Cold Spring

Harbor, NY. 

Myhr, B., Bowers, L., and Caspary, W.J. (1985). 

Assays for the induction of gene mutations at the

thymidine kinase locus in L5178Y mouse lymphoma

cells in culture. In Progress in Mutation Research: 

Evaluation of Short-term Tests for Carcinogens; 

Report of the International Programme on Chemical

Safety’s Collaborative Study on In vitro Assays (J. 

Ashby, F.J. de Serres, M. Draper, M. Ishidate, Jr., 

B.H. Margolin, B.E. Matter, and M.D. Shelby, 

Eds.), Vol. 5, pp. 555-568. Elsevier Science

Publishers, Amsterdam. 

National Cancer Institute (NCI) (1976). Guidelines

for Carcinogen Bioassay in Small Rodents. Technical

Report Series No. 1. NIH Publication No. 76-801. 

U.S. Department of Health, Education, and Welfare, 

Public Health Service, National Institutes of Health, 

Bethesda, MD. 

National Institute for Occupational Safety and Health

(NIOSH) (1990). National Occupational Exposure

Survey (1981-1983), unpublished provisional data as

of July 1, 1990. NIOSH, Cincinnati, OH. 

National Institutes of Health (NIH) (1978). Open

Formula Rat and Mouse Ration (NIH-07). 

Specification NIH-11-1335. U.S. Department of

Health, Education, and Welfare, Public Health

Service, National Institutes of Health, Bethesda, MD. 

Patty’s Industrial Hygiene and Toxicology (1981). 

3rd ed. (G.D. Clayton and F.E. Clayton, Eds.), 

pp. 2061-2119. John Wiley and Sons, New York. 

Rao, G.N. (1995). Husbandry procedures other than

dietary restriction for lowering body weight and

tumor/disease rates in Fischer 344 rats. In Dietary

Restriction: Implications for the Design and

Interpretation of Toxicity and Carcinogenicity Studies

(R.W. Hart, D.A. Neumann, and R.T. Robertson, 

Eds.), pp. 52-78. ILSI Press, Washington, DC. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 53 

Registry of Toxic Effects of Chemical Substances

(RTECS) (1991). National Institute for Occupational

Safety and Health, RTECS database available through

the National Library of Medicine MEDLARS System; 

1971, updated quarterly. 

Sadtler Standard Spectra. IR No. C1603. Sadtler

Research Laboratories, Philadelphia. 

Scheuplein, R.J. and Blank, I.H. (1971). 

Permeability of the skin. Physiol. Rev. 51, 702-747. 

Shirley, E. (1977). A non-parametric equivalent of

Williams’ test for contrasting increasing dose levels of

a treatment. Biometrics 33, 386-389. 

Straus, D.S. (1981). Somatic mutation, cellular

differentiation, and cancer causation. JNCI 67, 

233-241. 

Tarone, R.E. (1975). Tests for trend in life table

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 in rodents from in vitro

genetic toxicity assays. Science 236, 933-941. 

U.S. International Trade Commission (USITC) 

(1994). Synthetic Organic Chemicals. United States

Production and Sales, 1992. USITC Publication

2720. U.S. International Trade Commission, 

Washington, DC. 

Ward, J.M., Fox, J.G, Anver, M.R., Haines, D.C., 

George, C.V., Collins, M.J., Jr., Gorelick, P.L., 

Nagashima, K., Gonda, M.A., Gilden, R.V., 

Tully, J.G., Russell, R.J., Benveniste, R.E., 

Paster, B.J., Dewhirst, F.E., Donovan, J.C., 

Anderson, L.M., and Rice, J.M. (1994). Chronic

active hepatitis and associated liver tumors in mice

caused by a persistent bacterial infection with a novel

Helicobacter species. J. Natl. Cancer Inst. 86, 

1222-1227. 

Williams, D.A. (1971). A test for differences

between treatment means when several dose levels are

compared with a zero dose control. Biometrics 27, 

103-117. 

Williams, D.A. (1972). The comparison of several

dose levels with a zero dose control. Biometrics 28, 

519-531. 

Zeiger, E., Anderson, B., Haworth S., Lawlor, T., 

Mortelmans, K., and Speck, W. (1987). Salmonella

mutagenicity tests: III. Results from the testing of

255 chemicals. Environ. Mutagen. 9 (Suppl. 9), 

1-110. 

Zeiger, E., Haseman, J.K., Shelby, M.D., 

Margolin, B.H., and Tennant, R.W. (1990). 

Evaluation of four in vitro genetic toxicity tests for

predicting rodent carcinogenicity: Confirmation of

earlier results with 41 additional chemicals. Environ. 

Mol. Mutagen. 16 (Suppl. 18), 1-14. 
54 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

55 

APPENDIX A

SUMMARY OF LESIONS IN MALE RATS

IN THE 2-YEAR DERMAL STUDY

OF SODIUM XYLENESULFONATE

TABLE A1 Summary of the Incidence of Neoplasms in Male Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 57 

TABLE A2 Individual Animal Tumor Pathology of Male Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 62 

TABLE A3 Statistical Analysis of Primary Neoplasms in Male Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 88 

TABLE A4 Summary of the Incidence of Nonneoplastic Lesions in Male Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 92 
56 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfate, NTP TR 464 57 

TABLE A1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Moribund 28 25 22 30 

Natural deaths 15 8 19 10 

Survivors

Terminal sacrifice 7 17 9 10 

Animals examined microscopically 50 50 50 50 

Alimentary System

Intestine large, colon (50) (33) (41) (50) 

Intestine large, cecum (50) (33) (41) (50) 

Intestine small, duodenum (50) (33) (41) (50) 

Carcinoma, metastatic, kidney 1 (2%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Sarcoma, metastatic, mesentery 1 (2%) 

Intestine small, jejunum (50) (33) (41) (50) 

Carcinoma 1 (2%) 1 (2%) 

Leiomyosarcoma 1 (3%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Intestine small, ileum (50) (33) (41) (50) 

Liver (50) (45) (47) (50) 

Carcinoma, metastatic, kidney 1 (2%) 

Hepatocellular carcinoma 1 (2%) 

Histiocytic sarcoma 1 (2%) 1 (2%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (2%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Mesentery (8) (7) (9) (8) 

Carcinoma, metastatic, kidney 1 (14%) 1 (11%) 

Histiocytic sarcoma 1 (13%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (14%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (11%) 

Sarcoma 1 (11%) 

Oral mucosa (2) (1) (3) (6) 

Gingival, squamous cell carcinoma 1 (50%) 1 (17%) 

Pharyngeal, squamous cell carcinoma 1 (17%) 

Pancreas (50) (33) (41) (50) 

Carcinoma, metastatic, kidney 1 (3%) 1 (2%) 

Histiocytic sarcoma 1 (2%) 1 (2%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (3%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Mixed tumor benign 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

58 Sodium Xylenesulfonate, NTP TR 464 

TABLE A1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Alimentary System (continued) 

Stomach, forestomach (50) (33) (41) (50) 

Leiomyosarcoma 1 (2%) 

Stomach, glandular (50) (33) (41) (50) 

Leiomyosarcoma 1 (2%) 

Sarcoma, metastatic, mesentery 1 (2%) 

Cardiovascular System

Heart (50) (33) (41) (50) 

Endocrine System

Adrenal cortex (50) (33) (41) (50) 

Adenoma 1 (2%) 

Histiocytic sarcoma 1 (2%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Adrenal medulla (50) (33) (41) (50) 

Pheochromocytoma malignant 1 (2%) 

Pheochromocytoma benign 8 (16%) 5 (15%) 3 (7%) 9 (18%) 

Islets, pancreatic (50) (33) (41) (50) 

Adenoma 5 (10%) 3 (6%) 

Carcinoma 1 (3%) 1 (2%) 

Carcinoma, metastatic, kidney 1 (3%) 1 (2%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (3%) 

Pituitary gland (50) (33) (41) (50) 

Adenoma, multiple 1 (2%) 

Pars distalis, adenoma 34 (68%) 26 (79%) 30 (73%) 31 (62%) 

Pars distalis, adenoma, multiple 9 (18%) 1 (3%) 4 (10%) 5 (10%) 

Pars distalis, carcinoma 1 (2%) 

Pars distalis, craniopharyngioma 1 (2%) 

Thyroid gland (50) (33) (41) (50) 

Bilateral, C-cell, adenoma 1 (2%) 1 (2%) 

Bilateral, C-cell, carcinoma 1 (2%) 

C-cell, adenoma 8 (16%) 3 (9%) 1 (2%) 5 (10%) 

Follicular cell, carcinoma 1 (2%) 1 (2%) 1 (2%) 

General Body System

Peritoneum (3) (1) (2) 

Genital System

Epididymis (50) (33) (41) (50) 

Preputial gland (50) (33) (40) (50) 

Adenoma 1 (2%) 1 (3%) 

Carcinoma 1 (3%) 

Prostate (50) (33) (41) (50) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfate, NTP TR 464 59 

TABLE A1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Genital System (continued) 

Seminal vesicle (50) (33) (41) (50) 

Carcinoma, metastatic, kidney 1 (3%) 1 (2%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (3%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Testes (50) (33) (41) (50) 

Bilateral, interstitial cell, adenoma 11 (22%) 2 (6%) 5 (12%) 17 (34%) 

Interstitial cell, adenoma 13 (26%) 6 (18%) 8 (20%) 13 (26%) 

Hematopoietic System

Bone marrow (50) (32) (41) (50) 

Histiocytic sarcoma 1 (2%) 1 (2%) 

Lymph node (41) (25) (31) (41) 

Mediastinal, carcinoma, metastatic, kidney 1 (4%) 1 (3%) 

Mediastinal, histiocytic sarcoma 1 (3%) 1 (2%) 

Renal, carcinoma, metastatic, kidney 1 (3%) 

Lymph node, mandibular (50) (33) (40) (49) 

Histiocytic sarcoma 1 (3%) 

Lymph node, mesenteric (50) (33) (41) (50) 

Carcinoma, metastatic, kidney 1 (3%) 1 (2%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (3%) 

Spleen (50) (46) (47) (50) 

Fibroma 1 (2%) 

Histiocytic sarcoma 1 (2%) 1 (2%) 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 (2%) 

Thymus (46) (31) (41) (50) 

Thymoma benign 1 (2%) 

Integumentary System

Mammary gland (50) (32) (40) (50) 

Carcinoma 1 (3%) 

Fibroadenoma 1 (2%) 

Skin (50) (50) (50) (50) 

Basal cell adenoma 1 (2%) 

Keratoacanthoma 1 (2%) 1 (2%) 

Squamous cell papilloma 1 (2%) 1 (2%) 

Sebaceous gland, skin, site of application, 

carcinoma 1 (2%) 

Skin, site of application, basal cell adenoma 1 (2%) 1 (2%) 

Subcutaneous tissue, fibroma 1 (2%) 1 (2%) 3 (6%) 

Subcutaneous tissue, histiocytic sarcoma 1 (2%) 

Subcutaneous tissue, schwannoma malignant 1 (2%) 

Subcutaneous tissue, pinna, melanoma

malignant 1 (2%) 

Subcutaneous tissue, skin, site of

application, fibroma, multiple 1 (2%) 

Subcutaneous tissue, skin, site of

application, fibrous histiocytoma 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

60 Sodium Xylenesulfonate, NTP TR 464 

TABLE A1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Musculoskeletal System

Bone (50) (32) (41) (50) 

Osteosarcoma 1 (2%) 

Rib, osteosarcoma 1 (3%) 

Skeletal muscle (3) (2) 

Carcinoma, metastatic, kidney 1 (33%) 1 (50%) 

Histiocytic sarcoma 1 (50%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (33%) 

Osteosarcoma, metastatic, bone 1 (33%) 

Nervous System

Brain (50) (33) (41) (50) 

Astrocytoma malignant 1 (2%) 1 (3%) 

Carcinoma, metastatic, pituitary gland 1 (2%) 

Oligodendroglioma malignant 1 (2%) 

Respiratory System

Lung (50) (33) (41) (50) 

Alveolar/bronchiolar adenoma 1 (2%) 

Alveolar/bronchiolar carcinoma 2 (4%) 

Carcinoma, metastatic, kidney 1 (3%) 1 (2%) 

Carcinoma, metastatic, skin 1 (2%) 

Histiocytic sarcoma 1 (2%) 1 (2%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (3%) 

Osteosarcoma, metastatic, bone 1 (2%) 

Mediastinum, osteosarcoma, metastatic, bone 1 (3%) 

Special Senses System

Harderian gland (1) (1) (1) 

Squamous cell carcinoma, metastatic, oral

mucosa 1 (100%) 

Zymbal’s gland (2) (2) 

Adenoma 1 (50%) 

Carcinoma 1 (50%) 2 (100%) 

Urinary System

Kidney (50) (33) (41) (50) 

Adenoma, tubular 1 (2%) 

Carcinoma, metastatic, kidney 1 (3%) 

Sarcoma 1 (3%) 

Renal tubule, adenoma 1 (2%) 1 (2%) 

Renal tubule, carcinoma 1 (3%) 1 (2%) 

Urinary bladder (50) (33) (41) (50) 

Transitional epithelium, carcinoma 1 (2%) 

Transitional epithelium, papilloma 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

61 

c

Sodium Xylenesulfate, NTP TR 464 

TABLE A1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Systemic Lesions

Multiple organsb (50) (50) (50) (50) 

Histiocytic sarcoma 1 (2%) 1 (2%) 

Leukemia mononuclear 12 (24%) 24 (48%) 15 (30%) 25 (50%) 

Mesothelioma malignant 1 (2%) 

Neoplasm Summary

Total animals with primary neoplasmsc 50 45 46 49 

Total primary neoplasms 120 80 85 127 

Total animals with benign neoplasms 49 29 39 49 

Total benign neoplasms 98 47 58 92 

Total animals with malignant neoplasms 18 28 25 30 

Total malignant neoplasms 22 33 27 35 

Total animals with metastatic neoplasms 7 4 4 3 

Total metastatic neoplasms 42 33 21 22 

Total animals with malignant neoplasmsuncertain primary site 5 1 2 

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
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

62 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 

Vehicle Control

4444555555555555566666666 

Number of Days on Study 4499234455666889911113344 

1729746747057230566786958 

0000000000000000000000000 

Carcass ID Number 0013342104300403213331422 

8504849379126019465765301 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Intestine large, rectum +++++++++++++++++++++++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X

Intestine small, duodenum +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Intestine small, jejunum +++++++++++++++++++++++++ 

Carcinoma X

Mesothelioma malignant, metastatic, 

uncertain primary site X

Intestine small, ileum +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Liver +++++++++++++++++++++++++ 

Mesentery ++ + + 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Oral mucosa + 

Gingival, squamous cell carcinoma X

Pancreas +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Stomach, glandular +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Tooth

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Adenoma X

Adrenal medulla +++++++++++++++++++++++++ 

Pheochromocytoma benign X

Islets, pancreatic +++++++++++++++++++++++++ 

Adenoma X X X

Parathyroid gland +++++ M +++++++ M ++++++++ M ++ 

+: Tissue examined microscopically M: Missing tissue X: Lesion present

A: Autolysis precludes examination I: Insufficient tissue Blank: Not examined
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 63 

TABLE A2
#

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

6666666677777777777777777 

Number of Days on Study 5557888800001112223333333 

0282127712782354885555555 

0000000000000000000000000 Total

Carcass ID Number 2003114411232423240112445 Tissues/ 

7943176884206732523298150 Tumors

Alimentary System

Esophagus +++++++++++++++++++++++++ 50 

Intestine large, colon +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 2 

Intestine large, rectum +++++++++++++++++++++++++ 50 

Intestine large, cecum +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 1 

Intestine small, duodenum +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 2 

Intestine small, jejunum +++++++++++++++++++++++++ 50 

Carcinoma 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Intestine small, ileum +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 2 

Liver +++++++++++++++++++++++++ 50 

Mesentery + + ++ 8 

Mesothelioma malignant, metastatic, 

uncertain primary site X 3 

Oral mucosa + 2 

Gingival, squamous cell carcinoma 1 

Pancreas +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X 3 

Salivary glands +++++++++++++++++++++++++ 50 

Stomach, forestomach +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 2 

Stomach, glandular +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 2 

Tooth + 1 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 50 

Heart +++++++++++++++++++++++++ 50 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 50 

Adenoma 1 

Adrenal medulla +++++++++++++++++++++++++ 50 

Pheochromocytoma benign X X X X X X X 8 

Islets, pancreatic +++++++++++++++++++++++++ 50 

Adenoma X X 5 

Parathyroid gland +++++ M +++++++++++++++++++ 46 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

64 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2
#

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

4444555555555555566666666 

Number of Days on Study 4499234455666889911113344 

1729746747057230566786958 

0000000000000000000000000 

Carcass ID Number 0013342104300403213331422 

8504849379126019465765301 

Endocrine System (continued) 

Pituitary gland +++++++++++++++++++++++++ 

Adenoma, multiple X

Pars distalis, adenoma X XXXX XXXXXXXXX XXXXXX

Pars distalis, adenoma, multiple X

Pars distalis, carcinoma X

Thyroid gland +++++++++++++++++++++++++ 

Bilateral, C-cell, adenoma

C-cell, adenoma X X

Follicular cell, carcinoma

General Body System

Peritoneum + + + 

Mesothelioma malignant, metastatic, 

uncertain primary site X X X

Genital System

Epididymis +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X X

Preputial gland +++++++++++++++++++++++++ 

Adenoma

Prostate +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X

Seminal vesicle +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X

Testes +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X X X

Bilateral, interstitial cell, adenoma X X X

Interstitial cell, adenoma X X X X X

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Lymph node ++++++ + +++++++++++++ ++ 

Lymph node, mandibular +++++++++++++++++++++++++ 

Lymph node, mesenteric +++++++++++++++++++++++++ 

Spleen +++++++++++++++++++++++++ 

Thymus +++++++++++++++++++++++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 65 

TABLE A2
#

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

6666666677777777777777777 

Number of Days on Study 5557888800001112223333333 

0282127712782354885555555 

0000000000000000000000000 Total

Carcass ID Number 2003114411232423240112445 Tissues/ 

7943176884206732523298150 Tumors

Endocrine System (continued) 

Pituitary gland +++++++++++++++++++++++++ 50 

Adenoma, multiple 1 

Pars distalis, adenoma X X X X X X X X XXXX X X 34 

Pars distalis, adenoma, multiple X X X X XXX X 9 

Pars distalis, carcinoma 1 

Thyroid gland +++++++++++++++++++++++++ 50 

Bilateral, C-cell, adenoma X 1 

C-cell, adenoma X X X X X X 8 

Follicular cell, carcinoma X 1 

General Body System

Peritoneum 3 

Mesothelioma malignant, metastatic, 

uncertain primary site 3 

Genital System

Epididymis +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X X 5 

Preputial gland +++++++++++++++++++++++++ 50 

Adenoma X 1 

Prostate +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Seminal vesicle +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 2 

Testes +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X X 5 

Bilateral, interstitial cell, adenoma X X X X X X X X 11 

Interstitial cell, adenoma X X X XXX X X 13 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 50 

Lymph node +++ +++ +++++++ + ++ +++ 41 

Lymph node, mandibular +++++++++++++++++++++++++ 50 

Lymph node, mesenteric +++++++++++++++++++++++++ 50 

Spleen +++++++++++++++++++++++++ 50 

Thymus + M + M + + M ++++++ M +++++++++++ 46 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

66 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2
#

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

4444555555555555566666666 

Number of Days on Study 4499234455666889911113344 

1729746747057230566786958 

0000000000000000000000000 

Carcass ID Number 0013342104300403213331422 

8504849379126019465765301 

Integumentary System

Mammary gland

Fibroadenoma

+++++++++++++++++++++++++ 

X

Skin +++++++++++++++++++++++++ 

Sebaceous gland, skin, site of

application, carcinoma

Subcutaneous tissue, pinna, melanoma

malignant

Subcutaneous tissue, skin, site of

application, fibroma, multiple

X

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Nervous System

Brain +++++++++++++++++++++++++ 

Astrocytoma malignant

Carcinoma, metastatic, pituitary

gland

Peripheral nerve

Spinal cord

X

+ 

+ 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Carcinoma, metastatic, skin

Nose

+++++++++++++++++++++++++ 

X

X

X

+++++++++++++++++++++++++ 

Trachea +++++++++++++++++++++++++ 

Special Senses System

Eye

Harderian gland

+ + 

Urinary System

Kidney

Adenoma, tubular

Mesothelioma malignant, metastatic, 

uncertain primary site

Renal tubule, adenoma

Urinary bladder

Mesothelioma malignant, metastatic, 

uncertain primary site

Transitional epithelium, papilloma

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

X X

X

Systemic Lesions

Multiple organs

Leukemia mononuclear

+++++++++++++++++++++++++ 

X X X X

Mesothelioma malignant X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 67 

TABLE A2
#

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

6666666677777777777777777 

Number of Days on Study 5557888800001112223333333 

0282127712782354885555555 

0000000000000000000000000 Total

Carcass ID Number 2003114411232423240112445 

7943176884206732523298150 

Tissues/ 

Tumors

Integumentary System

Mammary gland

Fibroadenoma

+++++++++++++++++++++++++ 50 

1 

Skin +++++++++++++++++++++++++ 50 

Sebaceous gland, skin, site of

application, carcinoma

Subcutaneous tissue, pinna, melanoma

malignant

Subcutaneous tissue, skin, site of

application, fibroma, multiple X

X

1 

1 

1 

Musculoskeletal System

Bone +++++++++++++++++++++++++ 50 

Nervous System

Brain +++++++++++++++++++++++++ 50 

Astrocytoma malignant

Carcinoma, metastatic, pituitary

gland

Peripheral nerve

Spinal cord

X 1 

1 

1 

1 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Carcinoma, metastatic, skin

Nose

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

50 

1 

2 

1 

50 

Trachea +++++++++++++++++++++++++ 50 

Special Senses System

Eye

Harderian gland

 + 

+ 

3 

1 

Urinary System

Kidney

Adenoma, tubular

Mesothelioma malignant, metastatic, 

uncertain primary site

Renal tubule, adenoma

Urinary bladder

Mesothelioma malignant, metastatic, 

uncertain primary site

Transitional epithelium, papilloma

+++++++++++++++++++++++++ 

X

X

+++++++++++++++++++++++++ 

X

50 

1 

1 

1 

50 

3 

1 

Systemic Lesions

Multiple organs

Leukemia mononuclear

+++++++++++++++++++++++++ 

X X X XXXX X

50 

12 

Mesothelioma malignant 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

68 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

1333445555556666666666666 

Number of Days on Study 9469491244460000012334445 

6923116701751339993113556 

0000000000000000000000000 

Carcass ID Number 6989757895569895868576587 

9966260717463481815585335 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Intestine large, rectum +++++++++++++++++++++++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Intestine small, duodenum +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site

Intestine small, jejunum +++++++++++++++++++++++++ 

Leiomyosarcoma X

Mesothelioma malignant, metastatic, 

uncertain primary site

Intestine small, ileum +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site

Liver +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Hepatocellular carcinoma

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Mesentery + + + + + 

Carcinoma, metastatic, kidney X

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Mesothelioma malignant, metastatic, 

uncertain primary site

Oral mucosa

Pancreas +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Mesothelioma malignant, metastatic, 

uncertain primary site

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site

Stomach, glandular +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site

Tongue + 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 69 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

6677777777777777777777777 

Number of Days on Study 5700012333333333333333333 

8422206255555555555555555 

0000100000000000000000000 Total

Carcass ID Number 8959086755666667777788999 Tissues/ 

1780000129234783467929245 Tumors

Alimentary System

Esophagus ++++++++ 33 

Intestine large, colon ++++++++ 33 

Intestine large, rectum ++++++++ 33 

Intestine large, cecum ++++++++ 33 

Intestine small, duodenum ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Intestine small, jejunum ++++++++ 33 

Leiomyosarcoma 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Intestine small, ileum ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Liver + + + + + + + + + + + + + + + + + + + + 45 

Carcinoma, metastatic, kidney 1 

Hepatocellular carcinoma X 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Mesentery + + 7 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Oral mucosa + 1 

Pancreas ++++++++ 33 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Salivary glands ++++++++ 33 

Stomach, forestomach ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Stomach, glandular ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Tongue 1 

Cardiovascular System

Blood vessel ++++++++ 33 

Heart ++++++++ 33 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

70 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

1333445555556666666666666 

Number of Days on Study 9469491244460000012334445 

6923116701751339993113556 

Carcass ID Number

Endocrine System

Adrenal cortex

Adrenal medulla

Pheochromocytoma benign

Islets, pancreatic

Carcinoma

Carcinoma, metastatic, kidney

Leiomyosarcoma, metastatic, 

intestine small, jejunum

Parathyroid gland

Pituitary gland

Pars distalis, adenoma

Pars distalis, adenoma, multiple

Thyroid gland

C-cell, adenoma

General Body System

Peritoneum

Mesothelioma malignant, metastatic, 

uncertain primary site

0000000000000000000000000 

6989757895569895868576587 

9966260717463481815585335 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

X

X

+ + M ++++++ M ++++++++++++ M ++ 

+++++++++++++++++++++++++ 

XXXX X XXXX XXXXXXX XXX

+++++++++++++++++++++++++ 

X X X

Genital System

Epididymis

Mesothelioma malignant, metastatic, 

uncertain primary site

Preputial gland

Prostate

Mesothelioma malignant, metastatic, 

uncertain primary site

Seminal vesicle

Carcinoma, metastatic, kidney

Leiomyosarcoma, metastatic, 

intestine small, jejunum

Mesothelioma malignant, metastatic, 

uncertain primary site

Testes

Mesothelioma malignant, metastatic, 

uncertain primary site

Bilateral, interstitial cell, adenoma

Interstitial cell, adenoma

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

X

+++++++++++++++++++++++++ 

X

X X X X

Hematopoietic System

Bone marrow

Lymph node

Mediastinal, carcinoma, metastatic, 

kidney

Lymph node, mandibular

+++++++++++++++ M +++++++++ 

+ + + +++++ ++++++++ + + 

X

+++++++++++++++++++++++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 71 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

6677777777777777777777777 

Number of Days on Study 5700012333333333333333333 

8422206255555555555555555 

0000100000000000000000000 Total

Carcass ID Number 8959086755666667777788999 Tissues/ 

1780000129234783467929245 Tumors

Endocrine System

Adrenal cortex ++++++++ 33 

Adrenal medulla ++++++++ 33 

Pheochromocytoma benign X X X X 5 

Islets, pancreatic ++++++++ 33 

Carcinoma X 1 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Parathyroid gland ++++ M + + + 29 

Pituitary gland ++++++++ 33 

Pars distalis, adenoma X X X XXXX 26 

Pars distalis, adenoma, multiple X 1 

Thyroid gland ++++++++ 33 

C-cell, adenoma 3 

General Body System

Peritoneum + 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Genital System

Epididymis ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Preputial gland ++++++++ 33 

Prostate ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Seminal vesicle ++++++++ 33 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Testes ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Bilateral, interstitial cell, adenoma X 2 

Interstitial cell, adenoma X X 6 

Hematopoietic System

Bone marrow ++++++++ 32 

Lymph node +++++++ 25 

Mediastinal, carcinoma, metastatic, 

kidney 1 

Lymph node, mandibular ++++++++ 33 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

72 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

1333445555556666666666666 

Number of Days on Study 9469491244460000012334445 

6923116701751339993113556 

0000000000000000000000000 

Carcass ID Number 6989757895569895868576587 

9966260717463481815585335 

Hematopoietic System (continued) 

Lymph node, mesenteric

Carcinoma, metastatic, kidney

Leiomyosarcoma, metastatic, 

intestine small, jejunum

Spleen

Fibroma

+++++++++++++++++++++++++ 

X

X

+++++++++++++++++++++++++ 

Thymus +++++ M +++++ M +++++++++++++ 

Integumentary System

Mammary gland

Carcinoma

+++++++++ M +++++++++++++++ 

Skin +++++++++++++++++++++++++ 

Keratoacanthoma

Skin, site of application, basal

cell adenoma

Subcutaneous tissue, fibroma

Subcutaneous tissue, skin, site of

application, fibrous histiocytoma

X

Musculoskeletal System

Bone +++++++++++++++ M +++++++++ 

Rib, osteosarcoma

Skeletal muscle + + 

Carcinoma, metastatic, kidney

Leiomyosarcoma, metastatic, 

intestine small, jejunum

Osteosarcoma, metastatic, bone

X

X

Nervous System

Brain +++++++++++++++++++++++++ 

Astrocytoma malignant

Peripheral nerve

Spinal cord

 + 

+ 

Respiratory System

Lung

Carcinoma, metastatic, kidney

Leiomyosarcoma, metastatic, 

intestine small, jejunum

Mediastinum, osteosarcoma, 

metastatic, bone

Nose

+++++++++++++++++++++++++ 

X

X

+++++++++++++++ M +++++++++ 

Trachea +++++++++++++++++++++++++ 

Special Senses System

Eye

Harderian gland + 

+ + 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 73 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

6677777777777777777777777 

Number of Days on Study 5700012333333333333333333 

8422206255555555555555555 

0000100000000000000000000 Total

Carcass ID Number 8959086755666667777788999 Tissues/ 

1780000129234783467929245 Tumors

Hematopoietic System (continued) 

Lymph node, mesenteric ++++++++ 33 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Spleen + + + + + + + + + + + + + + + + + + + + + 46 

Fibroma X 1 

Thymus ++++++++ 31 

Integumentary System

Mammary gland ++++++++ 32 

Carcinoma X 1 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Keratoacanthoma X 1 

Skin, site of application, basal

cell adenoma X 1 

Subcutaneous tissue, fibroma 1 

Subcutaneous tissue, skin, site of

application, fibrous histiocytoma X 1 

Musculoskeletal System

Bone ++++++++ 32 

Rib, osteosarcoma X 1 

Skeletal muscle + 3 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Osteosarcoma, metastatic, bone X 1 

Nervous System

Brain ++++++++ 33 

Astrocytoma malignant X 1 

Peripheral nerve 1 

Spinal cord 1 

Respiratory System

Lung ++++++++ 33 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Mediastinum, osteosarcoma, 

metastatic, bone X 1 

Nose ++++++++ 32 

Trachea ++++++++ 33 

Special Senses System

Eye 2 

Harderian gland 1 
 

 

 

 

 

 

 

 

 

 

74 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

1333445555556666666666666 

Number of Days on Study 9469491244460000012334445 

6923116701751339993113556 

0000000000000000000000000 

Carcass ID Number 6989757895569895868576587 

9966260717463481815585335 

Urinary System

Kidney +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Sarcoma X

Renal tubule, carcinoma X

Urinary bladder +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 

Leukemia mononuclear X X X X X X X X
 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 75 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

6677777777777777777777777 

Number of Days on Study 5700012333333333333333333 

8422206255555555555555555 

0000100000000000000000000 Total

Carcass ID Number 8959086755666667777788999 Tissues/ 

1780000129234783467929245 Tumors

Urinary System

Kidney ++++++++ 33 

Carcinoma, metastatic, kidney 1 

Sarcoma 1 

Renal tubule, carcinoma 1 

Urinary bladder ++++++++ 33 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 50 

Leukemia mononuclear X X XXXXXXX X X XXXXX 24 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

76 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

2444455555555556666666666 

Number of Days on Study 5378823366777990012222344 

7977975828139466921222766 

1111111111111111111111111 

Carcass ID Number 3145433043012140332012423 

7380452216501969404477739 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Intestine large, rectum +++++++++++++++++++++++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Intestine small, duodenum +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Sarcoma, metastatic, mesentery X

Intestine small, jejunum +++++++++++++++++++++++++ 

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Intestine small, ileum +++++++++++++++++++++++++ 

Liver +++++++++++++++++++++++++ 

Histiocytic sarcoma

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Mesentery ++ +++ + + 

Carcinoma, metastatic, kidney X

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Sarcoma X

Oral mucosa + 

Pancreas +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Histiocytic sarcoma

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Mixed tumor benign X

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Leiomyosarcoma X

Stomach, glandular +++++++++++++++++++++++++ 

Leiomyosarcoma X

Sarcoma, metastatic, mesentery X

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Adrenal medulla +++++++++++++++++++++++++ 

Pheochromocytoma benign X X

Islets, pancreatic +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Parathyroid gland ++++++++++++++++++ M M +++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 77 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

6666666667777777777777777 

Number of Days on Study 5555677790000112333333333 

0239622350488586555555555 

1111111111111111111111111 Total

Carcass ID Number 1023124320412204001112344 Tissues/ 

5828883361515962372460109 Tumors

Alimentary System

Esophagus ++++++++++++++++ 41 

Intestine large, colon ++++++++++++++++ 41 

Intestine large, rectum ++++++++++++++++ 41 

Intestine large, cecum ++++++++++++++++ 41 

Intestine small, duodenum ++++++++++++++++ 41 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Sarcoma, metastatic, mesentery 1 

Intestine small, jejunum ++++++++++++++++ 41 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Intestine small, ileum ++++++++++++++++ 41 

Liver + + + + + + + + + + + + + + + + + + + + + + 47 

Histiocytic sarcoma X 1 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Mesentery ++ 9 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Sarcoma 1 

Oral mucosa ++ 3 

Pancreas ++++++++++++++++ 41 

Carcinoma, metastatic, kidney 1 

Histiocytic sarcoma X 1 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Mixed tumor benign 1 

Salivary glands ++++++++++++++++ 41 

Stomach, forestomach ++++++++++++++++ 41 

Leiomyosarcoma 1 

Stomach, glandular ++++++++++++++++ 41 

Leiomyosarcoma 1 

Sarcoma, metastatic, mesentery 1 

Cardiovascular System

Blood vessel ++++++++++++++++ 41 

Heart ++++++++++++++++ 41 

Endocrine System

Adrenal cortex ++++++++++++++++ 41 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Adrenal medulla ++++++++++++++++ 41 

Pheochromocytoma benign X 3 

Islets, pancreatic ++++++++++++++++ 41 

Carcinoma, metastatic, kidney 1 

Parathyroid gland ++++++++++++++++ 39 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

78 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

2444455555555556666666666 

Number of Days on Study 5378823366777990012222344 

7977975828139466921222766 

Carcass ID Number

1111

3145

7380

1

4

4

1 1

3 3

5 2

1 1

0 4

2 1

1 1

3 0

6 5

11111111111111 

12140332012423 

01969404477739 

Endocrine System (continued) 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X X X X XXXXXXXXXXXXX

Pars distalis, adenoma, multiple X X X

Thyroid gland +++++++++++++++++++++++++ 

C-cell, adenoma X

Follicular cell, carcinoma X

General Body System

None

Genital System

Coagulating gland

Epididymis +++++++++++++++++++++++++ 

Preputial gland +++++++++++++++++++++++++ 

Adenoma X

Carcinoma

Prostate +++++++++++++++++++++++++ 

Seminal vesicle +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Testes +++++++++++++++++++++++++ 

Bilateral, interstitial cell, adenoma X X

Interstitial cell, adenoma X X X X X

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Histiocytic sarcoma

Lymph node ++++++++++ + ++++ +++ 

Mediastinal, carcinoma, metastatic, 

kidney X

Mediastinal, histiocytic sarcoma

Renal, carcinoma, metastatic, kidney X

Lymph node, mandibular + + M ++++++++++++++++++++++ 

Histiocytic sarcoma

Lymph node, mesenteric +++++++++++++++++++++++++ 

Carcinoma, metastatic, kidney X

Spleen +++++++++++++++++++++++++ 

Histiocytic sarcoma

Leiomyosarcoma, metastatic, stomach, 

forestomach X

Thymus +++++++++++++++++++++++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 79 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

6666666667777777777777777 

Number of Days on Study 5555677790000112333333333 

0239622350488586555555555 

1111111111111111111111111 Total

Carcass ID Number 1023124320412204001112344 Tissues/ 

5828883361515962372460109 Tumors

Endocrine System (continued) 

Pituitary gland ++++++++++++++++ 41 

Pars distalis, adenoma X X X X XXXXXXX X X 30 

Pars distalis, adenoma, multiple X 4 

Thyroid gland ++++++++++++++++ 41 

C-cell, adenoma 1 

Follicular cell, carcinoma 1 

General Body System

None

Genital System

Coagulating gland + 1 

Epididymis ++++++++++++++++ 41 

Preputial gland +++ M ++++++++++++ 40 

Adenoma 1 

Carcinoma X 1 

Prostate ++++++++++++++++ 41 

Seminal vesicle ++++++++++++++++ 41 

Carcinoma, metastatic, kidney 1 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Testes ++++++++++++++++ 41 

Bilateral, interstitial cell, adenoma X X X 5 

Interstitial cell, adenoma X X X 8 

Hematopoietic System

Bone marrow ++++++++++++++++ 41 

Histiocytic sarcoma X 1 

Lymph node ++++++ ++ +++++ 31 

Mediastinal, carcinoma, metastatic, 

kidney 1 

Mediastinal, histiocytic sarcoma X 1 

Renal, carcinoma, metastatic, kidney 1 

Lymph node, mandibular ++++++++++++++++ 40 

Histiocytic sarcoma X 1 

Lymph node, mesenteric ++++++++++++++++ 41 

Carcinoma, metastatic, kidney 1 

Spleen + + + + + + + + + + + + + + + + + + + + + + 47 

Histiocytic sarcoma X 1 

Leiomyosarcoma, metastatic, stomach, 

forestomach 1 

Thymus ++++++++++++++++ 41 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

80 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

2444455555555556666666666 

Number of Days on Study 5378823366777990012222344 

7977975828139466921222766 

1111111111111111111111111 

Carcass ID Number 3145433043012140332012423 

7380452216501969404477739 

Integumentary System

Mammary gland

Skin

Keratoacanthoma

Squamous cell papilloma

Skin, site of application, basal

cell adenoma

Subcutaneous tissue, fibroma

Subcutaneous tissue, schwannoma

malignant

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

Musculoskeletal System

Bone

Osteosarcoma

Skeletal muscle

Carcinoma, metastatic, kidney

Histiocytic sarcoma

+++++++++++++++++++++++++ 

X

+ 

X

Nervous System

Brain

Oligodendroglioma malignant

Peripheral nerve

Spinal cord

+++++++++++++++++++++++++ 

X

Respiratory System

Lung

Carcinoma, metastatic, kidney

Histiocytic sarcoma

Osteosarcoma, metastatic, bone

Nose

Trachea

+++++++++++++++++++++++++ 

X

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Special Senses System

Eye

Zymbal’s gland

Adenoma

Carcinoma

+ 

X

+ 

Urinary System

Kidney

Renal tubule, carcinoma

Urinary bladder

Transitional epithelium, carcinoma

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Leukemia mononuclear

+++++++++++++++++++++++++ 

X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 81 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

6666666667777777777777777 

Number of Days on Study 5555677790000112333333333 

0239622350488586555555555 

1111111111111111111111111 Total

Carcass ID Number 1023124320412204001112344 Tissues/ 

5828883361515962372460109 Tumors

Integumentary System

Mammary gland + + M +++++++++++++ 40 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Keratoacanthoma X 1 

Squamous cell papilloma X 1 

Skin, site of application, basal

cell adenoma X 1 

Subcutaneous tissue, fibroma X 1 

Subcutaneous tissue, schwannoma

malignant 1 

Musculoskeletal System

Bone ++++++++++++++++ 41 

Osteosarcoma 1 

Skeletal muscle + 2 

Carcinoma, metastatic, kidney 1 

Histiocytic sarcoma X 1 

Nervous System

Brain ++++++++++++++++ 41 

Oligodendroglioma malignant 1 

Peripheral nerve + 1 

Spinal cord + 1 

Respiratory System

Lung ++++++++++++++++ 41 

Carcinoma, metastatic, kidney 1 

Histiocytic sarcoma X 1 

Osteosarcoma, metastatic, bone 1 

Nose ++++++++++++++++ 41 

Trachea ++++++++++++++++ 41 

Special Senses System

Eye + 2 

Zymbal’s gland + 2 

Adenoma X 1 

Carcinoma 1 

Urinary System

Kidney ++++++++++++++++ 41 

Renal tubule, carcinoma 1 

Urinary bladder ++++++++++++++++ 41 

Transitional epithelium, carcinoma X 1 

Systemic Lesions

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Histiocytic sarcoma X 1 

Leukemia mononuclear X XXX X X X XXX X X X 15 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

82 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

3444555555555555555666666 

Number of Days on Study 8238222234456666899123345 

2830045701761278204691822 

1111111111111111111111111 

Carcass ID Number 7577967987687895988969668 

4863160668051055371048358 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Intestine large, rectum +++++ M +++++++++++++++++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Intestine small, duodenum +++++++++++++++++++++++++ 

Intestine small, jejunum +++++++++++++++++++++++++ 

Carcinoma X

Mesothelioma malignant, metastatic, 

uncertain primary site X

Intestine small, ileum +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X

Liver +++++++++++++++++++++++++ 

Histiocytic sarcoma

Mesentery + + + 

Histiocytic sarcoma

Mesothelioma malignant, metastatic, 

uncertain primary site X

Oral mucosa + + 

Gingival, squamous cell carcinoma X

Pharyngeal, squamous cell carcinoma

Pancreas +++++++++++++++++++++++++ 

Histiocytic sarcoma

Mesothelioma malignant, metastatic, 

uncertain primary site X

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site

Stomach, glandular +++++++++++++++++++++++++ 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Histiocytic sarcoma

Adrenal medulla +++++++++++++++++++++++++ 

Pheochromocytoma malignant X

Pheochromocytoma benign X

Islets, pancreatic +++++++++++++++++++++++++ 

Adenoma X

Carcinoma X

Parathyroid gland ++++++++++++++++++ M +++++ M

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X XXX X X X XXXXXXX X

Pars distalis, adenoma, multiple X X

Pars distalis, craniopharyngioma
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 83 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

6666677777777777777777777 

Number of Days on Study 5667800111222333333333333 

8799722499668015555555555 

1111111111121111111111111 Total

Carcass ID Number 5578789969608755555666789 Tissues/ 

9394792927802261247179534 Tumors

Alimentary System

Esophagus +++++++++++++++++++++++++ 50 

Intestine large, colon +++++++++++++++++++++++++ 50 

Intestine large, rectum +++++++++++++++++++++++++ 49 

Intestine large, cecum +++++++++++++++++++++++++ 50 

Intestine small, duodenum +++++++++++++++++++++++++ 50 

Intestine small, jejunum +++++++++++++++++++++++++ 50 

Carcinoma 1 

Mesothelioma malignant, metastatic, 

uncertain primary site 1 

Intestine small, ileum +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site 1 

Liver +++++++++++++++++++++++++ 50 

Histiocytic sarcoma X 1 

Mesentery + ++ + + 8 

Histiocytic sarcoma X 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Oral mucosa ++ ++ 6 

Gingival, squamous cell carcinoma 1 

Pharyngeal, squamous cell carcinoma X 1 

Pancreas +++++++++++++++++++++++++ 50 

Histiocytic sarcoma X 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Salivary glands +++++++++++++++++++++++++ 50 

Stomach, forestomach +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Stomach, glandular +++++++++++++++++++++++++ 50 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 50 

Heart +++++++++++++++++++++++++ 50 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 50 

Histiocytic sarcoma X 1 

Adrenal medulla +++++++++++++++++++++++++ 50 

Pheochromocytoma malignant 1 

Pheochromocytoma benign XXX X X X X X 9 

Islets, pancreatic +++++++++++++++++++++++++ 50 

Adenoma X X 3 

Carcinoma 1 

Parathyroid gland + M ++++++++++++++ M +++++ M ++ 45 

Pituitary gland +++++++++++++++++++++++++ 50 

Pars distalis, adenoma X X X XXXX XXXXXXX X X 31 

Pars distalis, adenoma, multiple X X X 5 

Pars distalis, craniopharyngioma X 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

84 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

3444555555555555555666666 

Number of Days on Study 8238222234456666899123345 

2830045701761278204691822 

1111111111111111111111111 

Carcass ID Number 7577967987687895988969668 

4863160668051055371048358 

Endocrine System (continued) 

Thyroid gland +++++++++++++++++++++++++ 

Bilateral, C-cell, adenoma

Bilateral, C-cell, carcinoma

C-cell, adenoma X

Follicular cell, carcinoma

General Body System

Peritoneum + 

Mesothelioma malignant, metastatic, 

uncertain primary site X

Genital System

Coagulating gland

Epididymis +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X

Preputial gland +++++++++++++++++++++++++ 

Prostate +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X

Seminal vesicle +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site

Testes +++++++++++++++++++++++++ 

Mesothelioma malignant, metastatic, 

uncertain primary site X

Bilateral, interstitial cell, adenoma X X X X X

Interstitial cell, adenoma XXX X X X X

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Histiocytic sarcoma

Lymph node + +++++++++ +++ ++ +++++ 

Mediastinal, histiocytic sarcoma

Lymph node, mandibular +++++++++++++++++++++++++ 

Lymph node, mesenteric +++++++++++++++++++++++++ 

Spleen +++++++++++++++++++++++++ 

Histiocytic sarcoma

Mesothelioma malignant, metastatic, 

uncertain primary site

Thymus +++++++++++++++++++++++++ 

Thymoma benign X

Integumentary System

Mammary gland +++++++++++++++++++++++++ 

Skin +++++++++++++++++++++++++ 

Basal cell adenoma

Squamous cell papilloma

Subcutaneous tissue, fibroma X

Subcutaneous tissue, histiocytic

sarcoma
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 85 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

6666677777777777777777777 

Number of Days on Study 5667800111222333333333333 

8799722499668015555555555 

1111111111121111111111111 Total

Carcass ID Number 5578789969608755555666789 Tissues/ 

9394792927802261247179534 Tumors

Endocrine System (continued) 

Thyroid gland +++++++++++++++++++++++++ 50 

Bilateral, C-cell, adenoma X 1 

Bilateral, C-cell, carcinoma X 1 

C-cell, adenoma X X X X 5 

Follicular cell, carcinoma X 1 

General Body System

Peritoneum + 2 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Genital System

Coagulating gland + + 2 

Epididymis +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Preputial gland +++++++++++++++++++++++++ 50 

Prostate +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Seminal vesicle +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Testes +++++++++++++++++++++++++ 50 

Mesothelioma malignant, metastatic, 

uncertain primary site X 2 

Bilateral, interstitial cell, adenoma X X X X X X X X XXX X 17 

Interstitial cell, adenoma X X X X X X 13 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 50 

Histiocytic sarcoma X 1 

Lymph node ++ + ++ +++ +++++++++++++ 41 

Mediastinal, histiocytic sarcoma X 1 

Lymph node, mandibular ++++++++++++++ M ++++++++++ 49 

Lymph node, mesenteric +++++++++++++++++++++++++ 50 

Spleen +++++++++++++++++++++++++ 50 

Histiocytic sarcoma X 1 

Mesothelioma malignant, metastatic, 

uncertain primary site X 1 

Thymus +++++++++++++++++++++++++ 50 

Thymoma benign 1 

Integumentary System

Mammary gland +++++++++++++++++++++++++ 50 

Skin +++++++++++++++++++++++++ 50 

Basal cell adenoma X 1 

Squamous cell papilloma X 1 

Subcutaneous tissue, fibroma X X 3 

Subcutaneous tissue, histiocytic

sarcoma X 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

86 Sodium Xylenesulfonate, NTP TR 464 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

Number of Days on Study

3444555555555555555666666 

8238222234456666899123345 

2830045701761278204691822 

Carcass ID Number

1111111111111111111111111 

7577967987687895988969668 

4863160668051055371048358 

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Nervous System

Brain +++++++++++++++++++++++++ 

Respiratory System

Lung

Histiocytic sarcoma

Nose

Trachea

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Special Senses System

Eye

Harderian gland

Squamous cell carcinoma, metastatic, 

oral mucosa

Zymbal’s gland

Carcinoma

+ 

X

+ 

+ 

X

Urinary System

Kidney

Mesothelioma malignant, metastatic, 

uncertain primary site

Renal tubule, adenoma

Urinary bladder

Mesothelioma malignant, metastatic, 

uncertain primary site

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

X

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Leukemia mononuclear

+++++++++++++++++++++++++ 

X X XXX X X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 87 

TABLE A2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

Number of Days on Study

6666677777777777777777777 

5667800111222333333333333 

8799722499668015555555555 

Carcass ID Number

1111111111121111111111111 

5578789969608755555666789 

9394792927802261247179534 

Total

Tissues/ 

Tumors

Musculoskeletal System

Bone +++++++++++++++++++++++++ 50 

Nervous System

Brain +++++++++++++++++++++++++ 50 

Respiratory System

Lung

Histiocytic sarcoma

Nose

Trachea

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

1 

50 

50 

Special Senses System

Eye

Harderian gland

Squamous cell carcinoma, metastatic, 

oral mucosa

Zymbal’s gland

Carcinoma

+ 

X

1 

1 

1 

2 

2 

Urinary System

Kidney

Mesothelioma malignant, metastatic, 

uncertain primary site

Renal tubule, adenoma

Urinary bladder

Mesothelioma malignant, metastatic, 

uncertain primary site

+++++++++++++++++++++++++ 

X

X

+++++++++++++++++++++++++ 

X

50 

2 

1 

50 

2 

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Leukemia mononuclear

+++++++++++++++++++++++++ 

X

X X X X X X XXXXXXXXXXX

50 

1 

25 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

88 Sodium Xylenesulfonate, NTP TR 464 

TABLE A3 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Adrenal Medulla: Benign Pheochromocytoma

Overall ratea

Adjusted rateb

Terminal ratec

First incidence (days) 

Life table testd

Logistic regression testd

Fisher exact testd

8/50 (16%) 

44.8% 

1/7 (14%) 

554 

5/33 (15%) e 3/41 (7%) e 9/50 (18%) 

49.7% 

3/10 (30%) 

642 

P ̆0.482N

P ̆0.557 

P ̆0.500 

Adrenal Medulla: Benign or Malignant Pheochromocytoma

Overall rate 8/50 (16%) 

Adjusted rate 44.8% 

Terminal rate 1/7 (14%) 

First incidence (days) 554 

Life table test

Logistic regression test

Fisher exact test

5/33 (15%) e 3/41 (7%) e 10/50 (20%) 

51.0% 

3/10 (30%) 

561 

P ̆0.580N

P ̆0.429 

P ̆0.398 

Lung: Alveolar/bronchiolar Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

3/50 (6%) 

13.8% 

0/7 (0%) 

617 

0/33 (0%) e 0/41 (0%) e 0/50 (0%) 

0.0% 

0/10 (0%) 

—f

P ̆0.109N

P ̆0.120N

P ̆0.121N

Pancreatic Islets: Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

5/50 (10%) 

20.6% 

0/7 (0%) 

492 

0/33 (0%) e 0/41 (0%) e 3/50 (6%) 

16.5% 

1/10 (10%) 

556 

P ̆0.305N

P ̆0.354N

P ̆0.357N

Pancreatic Islets: Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

5/50 (10%) 

20.6% 

0/7 (0%) 

492 

1/33 (3%) e 0/41 (0%) e 4/50 (8%) 

18.8% 

1/10 (10%) 

556 

P ̆0.446N

P ̆0.494N

P ̆0.500N
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 89 

TABLE A3
#

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Pituitary Gland (Pars Distalis or Unspecified Site): Adenoma

Overall rate 44/50 (88%) 

Adjusted rate 97.5% 

Terminal rate 6/7 (86%) 

First incidence (days) 441 

Life table test

Logistic regression test

Fisher exact test

27/33 (82%)e 34/41 (83%)e 36/50 (72%) 

89.0% 

6/10 (60%) 

428 

P ̆0.080N

P ̆0.041N

P ̆0.039N

Pituitary Gland (Pars Distalis or Unspecified Site): Adenoma or Carcinoma

Overall rate 45/50 (90%) 27/33 (82%)e

Adjusted rate 97.5% 

Terminal rate 6/7 (86%) 

First incidence (days) 441 

Life table test

Logistic regression test

Fisher exact test

34/41 (83%)e 36/50 (72%) 

89.0% 

6/10 (60%) 

428 

P ̆0.066N

P ̆0.021N

P ̆0.020N

Preputial Gland: Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

1/50 (2%) 

6.7% 

0/7 (0%) 

707 

0/33 (0%) e 2/40 (5%) e 0/50 (0%) 

0.0% 

0/10 (0%) 

—

P ̆0.464N

P ̆0.478N

P ̆0.500N

Skin: Squamous Cell Papilloma, Keratoacanthoma, or Basal Cell Adenoma

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

Adjusted rate 0.0% 9.3% 

Terminal rate 0/7 (0%) 0/17 (0%) 

First incidence (days) — 658 

Life table test P ̆0.244 P ̆0.350 

Logistic regression test P ̆0.241 P ̆0.262 

Cochran-Armitage test P ̆0.237 d

Fisher exact test P ̆0.247 

3/50 (6%) 

19.6% 

1/9 (11%) 

666 

P ̆0.130 

P ̆0.116 

P ̆0.121 

2/50 (4%) 

16.0% 

1/10 (10%) 

726 

P ̆0.339 

P ̆0.313 

P ̆0.247 

Skin (Subcutaneous Tissue): Fibroma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

1/50 (2%) 

4.5% 

0/7 (0%) 

672 

P ̆0.165 

P ̆0.162 

P ̆0.163 

1/50 (2%) 

3.2% 

0/17 (0%) 

631 

P ̆0.743N

P ̆0.762N

P ̆0.753N

1/50 (2%) 

4.8% 

0/9 (0%) 

666 

P ̆0.745 

P ̆0.761 

P ̆0.753N

3/50 (6%) 

18.9% 

1/10 (10%) 

527 

P ̆0.367 

P ̆0.307 

P ̆0.309 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

90 Sodium Xylenesulfonate, NTP TR 464 

TABLE A3
#

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Skin (Subcutaneous Tissue): Fibroma or Fibrous Histiocytoma

Overall rate 1/50 (2%) 

Adjusted rate 4.5% 

Terminal rate 0/7 (0%) 

First incidence (days) 672 

Life table test P ̆0.242 

Logistic regression test P ̆0.235 

Cochran-Armitage test P ̆0.237 

Fisher exact test

2/50 (4%) 

7.1% 

0/17 (0%) 

631 

P ̆0.527 

P ̆0.502 

P ̆0.500 

1/50 (2%) 

4.8% 

0/9 (0%) 

666 

P ̆0.745 

P ̆0.761 

P ̆0.753N

3/50 (6%) 

18.9% 

1/10 (10%) 

527 

P ̆0.367 

P ̆0.307 

P ̆0.309 

Testes: Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

24/50 (48%) 

94.8% 

6/7 (86%) 

534 

8/33 (24%) e 13/41 (32%)e 30/50 (60%) 

95.7% 

9/10 (90%) 

428 

P ̆0.484 

P ̆0.137 

P ̆0.158 

Thyroid Gland (C-cell): Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

9/50 (18%) 

54.2% 

2/7 (29%) 

547 

3/33 (9%) e 1/41 (2%) e 6/50 (12%) 

37.4% 

2/10 (20%) 

568 

P ̆0.157N

P ̆0.255N

P ̆0.288N

Thyroid Gland (C-cell): Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

9/50 (18%) 

54.2% 

2/7 (29%) 

547 

3/33 (9%) e 1/41 (2%) e 7/50 (14%) 

45.2% 

3/10 (30%) 

568 

P ̆0.220N

P ̆0.351N

P ̆0.393N

All Organs: Mononuclear Cell Leukemia

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

12/50 (24%) 

50.5% 

1/7 (14%) 

547 

P ̆0.035 

P ̆0.016 

P ̆0.024 

24/50 (48%) 

83.8% 

13/17 (76%) 

491 

P ̆0.252 

P ̆0.011 

P ̆0.011 

15/50 (30%) 

78.3% 

6/9 (67%) 

477 

P ̆0.376 

P ̆0.310 

P ̆0.326 

25/50 (50%) 

100.0% 

10/10 (100%) 

541 

P ̆0.060 

P ̆0.005 

P ̆0.006 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

91 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE A3
#

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

All Organs: Benign Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

49/50 (98%) 

100.0% 

7/7 (100%) 

441 

29/50 (58%)e 39/50 (78%)e 49/50 (98%) 

100.0% 

10/10 (100%) 

428 

P ̆0.286N

P ̆0.633 

P ̆0.753N

All Organs: Malignant Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

20/50 (40%) 

63.8% 

1/7 (14%) 

527 

29/50 (58%)e 25/50 (50%)e 32/50 (64%) 

100.0% 

10/10 (100%) 

433 

P ̆0.139 

P ̆0.012 

P ̆0.014 

All Organs: Benign or Malignant Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

50/50 (100%) 

100.0% 

7/7 (100%) 

441 

45/50 (90%)e 46/50 (92%)e 49/50 (98%) 

100.0% 

10/10 (100%) 

428 

P ̆0.252N

P ̆0.571N

P ̆0.500N

a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for adrenal gland, lung, 

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 dosed group incidence are the P values corresponding to pairwise

comparisons between the controls and that dosed 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 lower incidence in a dose group is indicated by N. e Tissues (except skin) were examined microscopically only in those animals dying prior to terminal sacrifice or when it was observed to be abnormal at
 

necropsy; thus statistical comparisons with the controls are not applicable.
#

f	 Not applicable; no neoplasms in animal group
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

92 Sodium Xylenesulfonate, NTP TR 464 

TABLE A4 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Moribund 28 25 22 30 

Natural deaths 15 8 19 10 

Survivors

Terminal sacrifice 7 17 9 10 

Animals examined microscopically 50 50 50 50 

Alimentary System

Intestine large, colon (50) (33) (41) (50) 

Parasite metazoan 4 (8%) 1 (3%) 2 (5%) 3 (6%) 

Intestine large, rectum (50) (33) (41) (49) 

Parasite metazoan 2 (6%) 1 (2%) 2 (4%) 

Intestine large, cecum (50) (33) (41) (50) 

Inflammation, chronic active 1 (3%) 2 (5%) 1 (2%) 

Ulcer 1 (3%) 1 (2%) 1 (2%) 

Intestine small, duodenum (50) (33) (41) (50) 

Inflammation, chronic active 1 (2%) 1 (3%) 

Ulcer 1 (2%) 1 (3%) 

Intestine small, jejunum (50) (33) (41) (50) 

Diverticulum 1 (2%) 

Inflammation, chronic active 1 (2%) 1 (3%) 

Necrosis 1 (2%) 

Ulcer 1 (3%) 

Intestine small, ileum (50) (33) (41) (50) 

Inflammation, chronic active 2 (6%) 1 (2%) 

Parasite metazoan 1 (2%) 

Ulcer 1 (2%) 

Liver (50) (45) (47) (50) 

Angiectasis 1 (2%) 

Basophilic focus 3 (6%) 3 (6%) 6 (12%) 

Clear cell focus 3 (6%) 1 (2%) 

Degeneration, cystic 12 (24%) 4 (9%) 8 (17%) 9 (18%) 

Hematopoietic cell proliferation 1 (2%) 1 (2%) 2 (4%) 

Hepatodiaphragmatic nodule 6 (12%) 2 (4%) 3 (6%) 9 (18%) 

Inflammation, chronic 20 (40%) 9 (20%) 10 (21%) 16 (32%) 

Necrosis 8 (16%) 4 (9%) 4 (9%) 2 (4%) 

Pigmentation, hemosiderin 1 (2%) 

Bile duct, hyperplasia 43 (86%) 29 (64%) 32 (68%) 43 (86%) 

Central vein, thrombosis 1 (2%) 

Hepatocyte, hyperplasia 1 (2%) 1 (2%) 2 (4%) 

Hepatocyte, vacuolization cytoplasmic 21 (42%) 11 (24%) 16 (34%) 20 (40%) 

Hepatocyte, centrilobular, degeneration, 

fatty 2 (4%) 4 (9%) 

Hepatocyte, centrilobular, necrosis 1 (2%) 3 (7%) 

Mesentery (8) (7) (9) (8) 

Mineralization 3 (38%) 1 (14%) 2 (22%) 2 (25%) 

Artery, inflammation, chronic active 1 (14%) 

Fat, inflammation, chronic active 4 (50%) 3 (43%) 4 (44%) 2 (25%) 

Fat, necrosis 1 (11%) 

a Number of animals examined microscopically at the site and the number of animals with lesion
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 93 

TABLE A4 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Alimentary System (continued) 

Oral mucosa (2) (1) (3) (6) 

Gingival, dysplasia 1 (33%) 

Gingival, inflammation, chronic active 1 (50%) 1 (100%) 2 (67%) 4 (67%) 

Pancreas (50) (33) (41) (50) 

Inflammation, chronic active 1 (2%) 

Acinus, atrophy 25 (50%) 13 (39%) 16 (39%) 13 (26%) 

Acinus, hyperplasia 3 (6%) 2 (5%) 1 (2%) 

Artery, inflammation, chronic active 2 (5%) 

Stomach, forestomach (50) (33) (41) (50) 

Cyst 1 (2%) 1 (2%) 

Inflammation, chronic active 12 (24%) 11 (33%) 11 (27%) 13 (26%) 

Mineralization 1 (2%) 2 (5%) 

Perforation 1 (2%) 

Ulcer 8 (16%) 10 (30%) 10 (24%) 11 (22%) 

Epithelium, hyperplasia 17 (34%) 11 (33%) 10 (24%) 17 (34%) 

Stomach, glandular (50) (33) (41) (50) 

Inflammation, chronic active 3 (6%) 4 (12%) 1 (2%) 

Mineralization 9 (18%) 1 (3%) 5 (12%) 4 (8%) 

Necrosis 1 (3%) 1 (2%) 2 (4%) 

Ulcer 2 (4%) 3 (9%) 1 (2%) 

Epithelium, erosion 1 (2%) 4 (12%) 3 (7%) 2 (4%) 

Epithelium, hyperplasia 1 (3%) 

Tongue (1) 

Cyst 1 (100%) 

Tooth (1) 

Inflammation, chronic active 1 (100%) 

Cardiovascular System

Blood vessel (50) (33) (41) (50) 

Mineralization 8 (16%) 5 (12%) 2 (4%) 

Thrombosis 1 (2%) 

Heart (50) (33) (41) (50) 

Cardiomyopathy, chronic 29 (58%) 21 (64%) 27 (66%) 28 (56%) 

Inflammation, chronic active 1 (2%) 1 (3%) 1 (2%) 

Mineralization 7 (14%) 1 (3%) 4 (10%) 1 (2%) 

Atrium, thrombosis 4 (8%) 6 (18%) 6 (15%) 8 (16%) 

Endocrine System

Adrenal cortex (50) (33) (41) (50) 

Accessory adrenal cortical nodule 1 (2%) 1 (2%) 

Atrophy 1 (2%) 

Degeneration, fatty 34 (68%) 23 (70%) 29 (71%) 33 (66%) 

Hyperplasia 25 (50%) 6 (18%) 17 (41%) 17 (34%) 

Hypertrophy 2 (6%) 1 (2%) 

Necrosis 1 (2%) 

Adrenal medulla (50) (33) (41) (50) 

Hyperplasia 30 (60%) 10 (30%) 17 (41%) 25 (50%) 

Islets, pancreatic (50) (33) (41) (50) 

Hyperplasia 2 (6%) 5 (12%) 1 (2%) 

Parathyroid gland (46) (29) (39) (45) 

Hyperplasia 13 (28%) 3 (10%) 8 (21%) 5 (11%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

94 Sodium Xylenesulfonate, NTP TR 464 

TABLE A4 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Endocrine System (continued) 

Pituitary gland (50) (33) (41) (50) 

Angiectasis 38 (76%) 26 (79%) 30 (73%) 35 (70%) 

Hemorrhage 1 (3%) 

Mineralization 5 (10%) 1 (2%) 

Necrosis 1 (3%) 1 (2%) 

Pigmentation, hematoidin 2 (4%) 1 (3%) 1 (2%) 

Pars distalis, atrophy 1 (2%) 

Pars distalis, cyst 8 (16%) 5 (15%) 10 (24%) 14 (28%) 

Pars distalis, hyperplasia 12 (24%) 5 (15%) 7 (17%) 17 (34%) 

Pars distalis, pigmentation, hemosiderin 33 (66%) 20 (61%) 26 (63%) 35 (70%) 

Pars distalis, thrombosis 1 (3%) 1 (2%) 

Pars intermedia, cyst 2 (4%) 2 (6%) 3 (6%) 

Pars nervosa, cyst 1 (2%) 

Thyroid gland (50) (33) (41) (50) 

C-cell, hyperplasia 7 (14%) 8 (24%) 7 (17%) 12 (24%) 

Follicle, cyst 2 (4%) 1 (2%) 4 (8%) 

General Body System

None

Genital System

Coagulating gland (1) (2) 

Inflammation, chronic active 1 (100%) 2 (100%) 

Epididymis (50) (33) (41) (50) 

Atrophy 25 (50%) 10 (30%) 16 (39%) 25 (50%) 

Inflammation, chronic active 1 (3%) 

Mineralization 1 (2%) 2 (5%) 

Preputial gland (50) (33) (40) (50) 

Cyst 2 (4%) 

Hyperplasia 1 (2%) 1 (3%) 1 (3%) 2 (4%) 

Inflammation, chronic active 47 (94%) 27 (82%) 38 (95%) 46 (92%) 

Duct, cyst 1 (2%) 1 (3%) 3 (8%) 

Prostate (50) (33) (41) (50) 

Atrophy 1 (2%) 

Cyst 3 (6%) 

Inflammation, chronic active 49 (98%) 32 (97%) 38 (93%) 47 (94%) 

Mineralization 1 (2%) 

Seminal vesicle (50) (33) (41) (50) 

Atrophy 3 (6%) 7 (21%) 11 (27%) 13 (26%) 

Inflammation, chronic active 2 (4%) 1 (3%) 

Mineralization 1 (2%) 

Testes (50) (33) (41) (50) 

Artery, inflammation, chronic 10 (20%) 2 (6%) 8 (20%) 9 (18%) 

Germinal epithelium, atrophy 20 (40%) 6 (18%) 19 (46%) 13 (26%) 

Interstitial cell, hyperplasia 20 (40%) 13 (39%) 11 (27%) 14 (28%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 95 

TABLE A4 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Hematopoietic System

Bone marrow (50) (32) (41) (50) 

Hyperplasia 25 (50%) 13 (41%) 16 (39%) 38 (76%) 

Lymph node (41) (25) (31) (41) 

Lumbar, hyperplasia, lymphoid 1 (4%) 

Mediastinal, atrophy 1 (2%) 

Mediastinal, ectasia 1 (2%) 

Mediastinal, infiltration cellular, 

histiocyte 1 (4%) 

Mediastinal, pigmentation, hemosiderin 37 (90%) 23 (92%) 27 (87%) 36 (88%) 

Renal, ectasia 2 (6%) 1 (2%) 

Renal, erythrophagocytosis 1 (2%) 

Renal, pigmentation, hemosiderin 1 (3%) 1 (2%) 

Lymph node, mesenteric (50) (33) (41) (50) 

Atrophy 1 (2%) 

Ectasia 1 (2%) 3 (9%) 2 (5%) 6 (12%) 

Inflammation, granulomatous 1 (2%) 

Spleen (50) (46) (47) (50) 

Fibrosis 6 (12%) 4 (9%) 8 (16%) 

Hematopoietic cell proliferation 12 (24%) 6 (13%) 7 (15%) 10 (20%) 

Pigmentation, hemosiderin 4 (8%) 1 (2%) 4 (8%) 

Lymphoid follicle, depletion cellular 1 (2%) 

Red pulp, depletion cellular 2 (4%) 

Integumentary System

Mammary gland (50) (32) (40) (50) 

Hyperplasia, cystic 42 (84%) 26 (81%) 35 (88%) 41 (82%) 

Inflammation, chronic active 1 (2%) 

Duct, cyst 1 (3%) 3 (8%) 2 (4%) 

Skin (50) (50) (50) (50) 

Hyperkeratosis 1 (2%) 

Epidermis, skin, site of application, 

hyperplasia 1 (2%) 1 (2%) 2 (4%) 

Sebaceous gland, skin, site of application, 

hyperplasia 1 (2%) 1 (2%) 

Skin, site of application, inflammation, 

chronic active 1 (2%) 

Subcutaneous tissue, inflammation, chronic

active 1 (2%) 

Musculoskeletal System

Bone (50) (32) (41) (50) 

Fibrous osteodystrophy 10 (20%) 1 (3%) 9 (22%) 8 (16%) 

Femur, osteosclerosis 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

96 Sodium Xylenesulfonate, NTP TR 464 

TABLE A4 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Nervous System

Brain (50) (33) (41) (50) 

Hemorrhage, acute 1 (3%) 4 (8%) 

Hydrocephalus 11 (22%) 12 (36%) 9 (22%) 5 (10%) 

Mineralization 1 (2%) 

Necrosis 1 (2%) 

Spinal cord (1) (1) (1) 

Axon, degeneration 1 (100%) 1 (100%) 

Respiratory System

Lung (50) (33) (41) (50) 

Congestion, acute 10 (20%) 9 (27%) 17 (41%) 6 (12%) 

Inflammation, chronic active 3 (6%) 1 (3%) 2 (5%) 3 (6%) 

Inflammation, granulomatous 2 (4%) 1 (2%) 

Inflammation, suppurative 1 (2%) 1 (2%) 

Mineralization 7 (14%) 5 (12%) 

Necrosis 1 (2%) 

Thrombosis 1 (2%) 

Alveolar epithelium, hyperplasia 3 (6%) 2 (6%) 1 (2%) 3 (6%) 

Alveolus, hemorrhage, acute 3 (6%) 1 (3%) 1 (2%) 

Alveolus, infiltration cellular, histiocyte 24 (48%) 10 (30%) 16 (39%) 16 (32%) 

Interstitium, inflammation, chronic active 7 (14%) 4 (10%) 3 (6%) 

Nose (50) (32) (41) (50) 

Inflammation, chronic active 5 (10%) 8 (25%) 4 (10%) 5 (10%) 

Polyp inflammatory 1 (2%) 

Thrombosis 1 (3%) 1 (2%) 

Nasolacrimal duct, inflammation, suppurative 5 (10%) 3 (9%) 3 (7%) 3 (6%) 

Special Senses System

Eye (3) (2) (2) (1) 

Anterior chamber, hemorrhage, acute 1 (33%) 

Cornea, inflammation, suppurative 1 (33%) 

Lens, cataract 2 (67%) 1 (50%) 1 (50%) 1 (100%) 

Retina, degeneration 2 (67%) 2 (100%) 1 (50%) 1 (100%) 

Harderian gland (1) (1) (1) 

Inflammation, chronic 1 (100%) 1 (100%) 

Urinary System

Kidney (50) (33) (41) (50) 

Accumulation, hyaline droplet 1 (2%) 

Infarct 2 (4%) 

Mineralization 4 (8%) 3 (9%) 2 (5%) 1 (2%) 

Nephropathy, chronic 50 (100%) 32 (97%) 41 (100%) 50 (100%) 

Cortex, cyst 8 (16%) 1 (3%) 1 (2%) 10 (20%) 

Urinary bladder (50) (33) (41) (50) 

Inflammation, chronic active 1 (2%) 

Mineralization 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

97 

APPENDIX B

SUMMARY OF LESIONS IN FEMALE RATS

IN THE 2-YEAR DERMAL STUDY

OF SODIUM XYLENESULFONATE

TABLE B1 Summary of the Incidence of Neoplasms in Female Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 99 

TABLE B2 Individual Animal Tumor Pathology of Female Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 102 

TABLE B3 Statistical Analysis of Primary Neoplasms in Female Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 118 

TABLE B4 Summary of the Incidence of Nonneoplastic Lesions in Female Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 122 
98 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 99 

TABLE B1 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Accidental death 1 

Moribund 14 10 15 12 

Natural deaths 13 24 18 22 

Survivors

Died last week of the study 1 

Terminal sacrifice 22 15 17 16 

Animals examined microscopically 50 50 50 50 

Alimentary System

Esophagus (50) (35) (33) (50) 

Periesophageal tissue, lipoma 1 (3%) 1 (3%) 

Intestine small, duodenum (50) (35) (33) (50) 

Liver (50) (35) (33) (50) 

Mesentery (2) (4) (4) (4) 

Oral mucosa (1) (2) 

Gingival, squamous cell carcinoma 1 (100%) 

Pancreas (50) (35) (33) (50) 

Stomach, glandular (50) (35) (33) (50) 

Cardiovascular System

Heart (50) (35) (33) (50) 

Endocrine System

Adrenal cortex (50) (35) (33) (50) 

Adenoma 1 (2%) 1 (3%) 1 (2%) 

Adrenal medulla (50) (35) (33) (50) 

Pheochromocytoma benign 1 (2%) 1 (3%) 2 (4%) 

Islets, pancreatic (50) (35) (33) (50) 

Adenoma 1 (3%) 

Carcinoma 1 (2%) 

Pituitary gland (50) (35) (33) (50) 

Pars distalis, adenoma 29 (58%) 16 (46%) 19 (58%) 27 (54%) 

Pars distalis, adenoma, multiple 5 (10%) 1 (3%) 2 (4%) 

Pars distalis, carcinoma 1 (2%) 

Pars intermedia, adenoma 1 (2%) 

Thyroid gland (50) (35) (33) (50) 

C-cell, adenoma 10 (20%) 1 (3%) 2 (6%) 7 (14%) 

C-cell, adenoma, multiple 1 (2%) 

Follicular cell, adenoma 1 (2%) 2 (6%) 1 (2%) 

Follicular cell, carcinoma 1 (2%) 

General Body System

None
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

100 Sodium Xylenesulfonate, NTP TR 464 

TABLE B1
#

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Genital System

Clitoral gland (49) (50) (50) (50) 

Adenoma 1 (2%) 3 (6%) 3 (6%) 

Carcinoma 1 (2%) 2 (4%) 

Bilateral, adenoma 1 (2%) 

Ovary (50) (35) (33) (50) 

Uterus (50) (35) (33) (50) 

Fibroma 1 (2%) 

Polyp stromal 2 (4%) 1 (3%) 1 (3%) 3 (6%) 

Sarcoma stromal 1 (3%) 1 (3%) 

Vagina (1) (1) (1) 

Squamous cell carcinoma 1 (100%) 

Hematopoietic System

Bone marrow (50) (35) (33) (48) 

Lymph node (45) (32) (27) (43) 

Lymph node, mandibular (50) (35) (33) (50) 

Lymph node, mesenteric (49) (35) (33) (48) 

Spleen (50) (35) (33) (50) 

Thymus (49) (35) (31) (49) 

Integumentary System

Mammary gland (50) (35) (33) (50) 

Carcinoma 1 (2%) 1 (3%) 1 (2%) 

Fibroadenoma 12 (24%) 4 (11%) 8 (24%) 6 (12%) 

Fibroadenoma, multiple 4 (8%) 3 (6%) 

Skin (50) (50) (50) (50) 

Pinna, squamous cell papilloma 1 (2%) 

Subcutaneous tissue, lipoma 1 (2%) 

Subcutaneous tissue, sarcoma 1 (2%) 

Subcutaneous tissue, schwannoma malignant 1 (2%) 

Musculoskeletal System 

Bone (50) (35) (33) (48) 

Humerus, osteosarcoma 1 (2%) 

Nervous System

Brain (50) (35) (33) (50) 

Astrocytoma malignant 1 (2%) 1 (3%) 

Carcinoma, metastatic, pituitary gland 1 (2%) 

Respiratory System

Lung (50) (35) (33) (50) 

Alveolar/bronchiolar adenoma 1 (2%) 

Osteosarcoma, metastatic, bone 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 101 

TABLE B1
#

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Special Senses System

Zymbal’s gland

Carcinoma

 (1) 

 1 (100%) 

Urinary System

Kidney

Lipoma

Urinary bladder

(50) 

(50) 

(35) 

 1 (3%) 

(35) 

(33) 

(33) 

(50) 

(50) 

Systemic Lesions

Multiple organsb

Leukemia mononuclear

Mesothelioma benign

(50) 

 17 (34%) 

(50) 

 5 (10%) 

(50) 

 9 (18%) 

(50) 

 10 (20%) 

 1 (2%) 

Neoplasm Summary

Total animals with primary neoplasmsc

Total primary neoplasms

Total animals with benign neoplasms

Total benign neoplasms

Total animals with malignant neoplasms

Total malignant neoplasms

Total animals with metastatic neoplasms

Total metastatic neoplasms

45 

93 

40 

69 

23 

24 

2 

2 

25 

40 

23 

32 

8 

8 

27 

50 

26 

37 

11 

13 

41 

70 

33 

55 

15 

15 

a

b

c

Number of animals examined microscopically at the site and the number of animals with neoplasm

Number of animals with any tissue examined microscopically

Primary neoplasms: all neoplasms except metastatic neoplasms
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

102 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 

Vehicle Control

3344444444555566666666667 

Number of Days on Study 0902288899447902455789990 

3380005800585159667422388 

2222222222222222222222222 

Carcass ID Number 3232241214241021211040333 

5781817347620726023268602 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Intestine large, rectum +++++++++++++++++++++++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Intestine small, duodenum +++++++++++++++++++++++++ 

Intestine small, jejunum +++++++++++++++++++++++++ 

Intestine small, ileum +++++++++++++++++++++++++ 

Liver +++++++++++++++++++++++++ 

Mesentery + 

Oral mucosa

Gingival, squamous cell carcinoma

Pancreas +++++++++++++++++++++++++ 

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Stomach, glandular +++++++++++++++++++++++++ 

Tooth

Cardiovascular System

Blood vessel

Heart

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Adenoma

Adrenal medulla +++++++++++++++++++++++++ 

Pheochromocytoma benign

Islets, pancreatic +++++++++++++++++++++++++ 

Carcinoma X

Parathyroid gland ++++++++++++++++++ M ++++++ 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X X X XXXXXXXX XXX X

Pars distalis, adenoma, multiple X

Pars distalis, carcinoma X

Thyroid gland +++++++++++++++++++++++++ 

C-cell, adenoma X X

Follicular cell, adenoma

General Body System

None

Genital System

Clitoral gland +++++++ M +++++++++++++++++ 

Adenoma

Ovary +++++++++++++++++++++++++ 

Uterus +++++++++++++++++++++++++ 

Fibroma

Polyp stromal X

Vagina + 

+: Tissue examined microscopically M: Missing tissue X: Lesion present

A: Autolysis precludes examination I: Insufficient tissue Blank: Not examined
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 103 

TABLE B2
#

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

7777777777777777777777777 

Number of Days on Study 1133333333333333333333333 

8926666666666666666666666 

2222222222222222222222222 Total

Carcass ID Number 1440000001112223333344445 Tissues/ 

1901345695894591347934580 Tumors

Alimentary System

Esophagus +++++++++++++++++++++++++ 50 

Intestine large, colon +++++++++++++++++++++++++ 50 

Intestine large, rectum +++++++++++++++++++++++++ 50 

Intestine large, cecum +++++++++++++++++++++++++ 50 

Intestine small, duodenum +++++++++++++++++++++++++ 50 

Intestine small, jejunum +++++++++++++++++++++++++ 50 

Intestine small, ileum +++++++++++++++++++++++++ 50 

Liver +++++++++++++++++++++++++ 50 

Mesentery + 2 

Oral mucosa + 1 

Gingival, squamous cell carcinoma X 1 

Pancreas +++++++++++++++++++++++++ 50 

Salivary glands +++++++++++++++++++++++++ 50 

Stomach, forestomach +++++++++++++++++++++++++ 50 

Stomach, glandular +++++++++++++++++++++++++ 50 

Tooth + 1 

Cardiovascular System

Blood vessel

Heart

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

50 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 50 

Adenoma X 1 

Adrenal medulla +++++++++++++++++++++++++ 50 

Pheochromocytoma benign X 1 

Islets, pancreatic +++++++++++++++++++++++++ 50 

Carcinoma 1 

Parathyroid gland ++++++++++++++++ M ++++++++ 48 

Pituitary gland +++++++++++++++++++++++++ 50 

Pars distalis, adenoma XXXXXXXXXX X X X X 29 

Pars distalis, adenoma, multiple X X X X 5 

Pars distalis, carcinoma 1 

Thyroid gland +++++++++++++++++++++++++ 50 

C-cell, adenoma X X X X X X X X 10 

Follicular cell, adenoma X 1 

General Body System

None

Genital System

Clitoral gland +++++++++++++++++++++++++ 49 

Adenoma X 1 

Ovary +++++++++++++++++++++++++ 50 

Uterus +++++++++++++++++++++++++ 50 

Fibroma X 1 

Polyp stromal X 2 

Vagina 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

104 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2
#

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

Number of Days on Study

3344444444555566666666667 

0902288899447902455789990 

3380005800585159667422388 

Carcass ID Number

2222222222222222222222222 

3232241214241021211040333 

5781817347620726023268602 

Hematopoietic System

Bone marrow

Lymph node

Lymph node, mandibular

Lymph node, mesenteric

Spleen

Thymus

+++++++++++++++++++++++++ 

+ + +++++++ +++++++++++++ 

+++++++++++++++++++++++++ 

++++++++++++++++++++++ M ++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Integumentary System

Mammary gland

Carcinoma

Fibroadenoma

Fibroadenoma, multiple

Skin

Pinna, squamous cell papilloma

Subcutaneous tissue, schwannoma

malignant

+++++++++++++++++++++++++ 

X X X X

+++++++++++++++++++++++++ 

X

Musculoskeletal System

Bone

Humerus, osteosarcoma

+++++++++++++++++++++++++ 

X

Nervous System

Brain

Astrocytoma malignant

Carcinoma, metastatic, pituitary

gland

+++++++++++++++++++++++++ 

X

X

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Osteosarcoma, metastatic, bone

Nose

Trachea

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Special Senses System

Eye + + + 

Urinary System

Kidney

Urinary bladder

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs

Leukemia mononuclear

+++++++++++++++++++++++++ 

X X X X X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 105 

TABLE B2
#

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

Number of Days on Study

7777777777777777777777777 

1133333333333333333333333 

8926666666666666666666666 

Carcass ID Number

2222222222222222222222222 

1440000001112223333344445 

1901345695894591347934580 

Total

Tissues/ 

Tumors

Hematopoietic System

Bone marrow

Lymph node

Lymph node, mandibular

Lymph node, mesenteric

Spleen

Thymus

+++++++++++++++++++++++++ 

++++++++++++++++++ +++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

++++++++++++++++++++ M ++++ 

50 

45 

50 

49 

50 

49 

Integumentary System

Mammary gland

Carcinoma

Fibroadenoma

Fibroadenoma, multiple

Skin

Pinna, squamous cell papilloma

Subcutaneous tissue, schwannoma

malignant

+++++++++++++++++++++++++ 

X

X X X X X X X X

X X X X

+++++++++++++++++++++++++ 

X

50 

1 

12 

4 

50 

1 

1 

Musculoskeletal System

Bone

Humerus, osteosarcoma

+++++++++++++++++++++++++ 50 

1 

Nervous System

Brain

Astrocytoma malignant

Carcinoma, metastatic, pituitary

gland

+++++++++++++++++++++++++ 50 

1 

1 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Osteosarcoma, metastatic, bone

Nose

Trachea

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

1 

1 

50 

50 

Special Senses System

Eye 3 

Urinary System

Kidney

Urinary bladder

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

50 

Systemic Lesions

Multiple organs

Leukemia mononuclear

+++++++++++++++++++++++++ 

X X X X X X X X X X X

50 

17 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

106 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

Number of Days on Study

2233344445555555555556666 

0706844892223445568892234 

1717667675887170932502603 

Carcass ID Number

2222222222222222222222222 

8587876967598879998567596 

5236830777521496516110782 

Alimentary System

Esophagus

Periesophageal tissue, lipoma

Intestine large, colon

Intestine large, rectum

Intestine large, cecum

Intestine small, duodenum

Intestine small, jejunum

Intestine small, ileum

Liver

Mesentery

Pancreas

Salivary glands

Stomach, forestomach

Stomach, glandular

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

 ++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Adenoma X

Adrenal medulla +++++++++++++++++++++++++ 

Islets, pancreatic +++++++++++++++++++++++++ 

Parathyroid gland ++++++++++++++++++++ M ++++ 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X X X XXXXX X X X

Pars distalis, adenoma, multiple

Thyroid gland +++++++++++++++++++++++++ 

C-cell, adenoma

Follicular cell, adenoma

General Body System

None

Genital System

Clitoral gland +++++++++++++++++++++++++ 

Adenoma X X

Ovary +++++++++++++++++++++++++ 

Uterus +++++++++++++++++++++++++ 

Polyp stromal X

Sarcoma stromal X

Vagina
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 107 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

6666677777777777777777777 

Number of Days on Study 4446900233333333333333333 

4556418256666666666666666 

2222322222222222222222222 Total

Carcass ID Number 5775086855566666777889999 Tissues/ 

6458023793445689128090349 Tumors

Alimentary System

Esophagus ++++++++++ 35 

Periesophageal tissue, lipoma 1 

Intestine large, colon ++++++++++ 35 

Intestine large, rectum ++++++++++ 35 

Intestine large, cecum ++++++++++ 35 

Intestine small, duodenum ++++++++++ 35 

Intestine small, jejunum ++++++++++ 35 

Intestine small, ileum ++++++++++ 35 

Liver ++++++++++ 35 

Mesentery + + 4 

Pancreas ++++++++++ 35 

Salivary glands ++++++++++ 35 

Stomach, forestomach ++++++++++ 35 

Stomach, glandular ++++++++++ 35 

Cardiovascular System

Blood vessel ++++++++++ 35 

Heart ++++++++++ 35 

Endocrine System

Adrenal cortex ++++++++++ 35 

Adenoma 1 

Adrenal medulla ++++++++++ 35 

Islets, pancreatic ++++++++++ 35 

Parathyroid gland ++++++ M + M + 32 

Pituitary gland ++++++++++ 35 

Pars distalis, adenoma X X X X X 16 

Pars distalis, adenoma, multiple X 1 

Thyroid gland ++++++++++ 35 

C-cell, adenoma X 1 

Follicular cell, adenoma X X 2 

General Body System

None

Genital System

Clitoral gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adenoma X 3 

Ovary ++++++++++ 35 

Uterus ++++++++++ 35 

Polyp stromal 1 

Sarcoma stromal 1 

Vagina + 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

108 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

2233344445555555555556666 

Number of Days on Study 0706844892223445568892234 

1717667675887170932502603 

2222222222222222222222222 

Carcass ID Number 8587876967598879998567596 

5236830777521496516110782 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Lymph node +++++++++++ ++++++ +++++ 

Lymph node, mandibular +++++++++++++++++++++++++ 

Lymph node, mesenteric +++++++++++++++++++++++++ 

Spleen +++++++++++++++++++++++++ 

Thymus +++++++++++++++++++++++++ 

Integumentary System

Mammary gland +++++++++++++++++++++++++ 

Carcinoma X

Fibroadenoma X

Skin +++++++++++++++++++++++++ 

Subcutaneous tissue, lipoma

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Skeletal muscle + 

Nervous System

Brain +++++++++++++++++++++++++ 

Astrocytoma malignant X

Respiratory System

Lung +++++++++++++++++++++++++ 

Nose +++++++++++++++++++++++++ 

Trachea +++++++++++++++++++++++++ 

Special Senses System

Eye + + 

Urinary System

Kidney +++++++++++++++++++++++++ 

Lipoma X

Urinary bladder +++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 

Leukemia mononuclear X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 109 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 60 mg/kg

(continued) 

6666677777777777777777777 

Number of Days on Study 4446900233333333333333333 

4556418256666666666666666 

2222322222222222222222222 Total

Carcass ID Number 5775086855566666777889999 Tissues/ 

6458023793445689128090349 Tumors

Hematopoietic System

Bone marrow ++++++++++ 35 

Lymph node ++++++++++ 32 

Lymph node, mandibular ++++++++++ 35 

Lymph node, mesenteric ++++++++++ 35 

Spleen ++++++++++ 35 

Thymus ++++++++++ 35 

Integumentary System

Mammary gland ++++++++++ 35 

Carcinoma 1 

Fibroadenoma X X X 4 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Subcutaneous tissue, lipoma X 1 

Musculoskeletal System

Bone ++++++++++ 35 

Skeletal muscle 1 

Nervous System

Brain ++++++++++ 35 

Astrocytoma malignant 1 

Respiratory System

Lung ++++++++++ 35 

Nose ++++++++++ 35 

Trachea ++++++++++ 35 

Special Senses System

Eye 2 

Urinary System

Kidney ++++++++++ 35 

Lipoma 1 

Urinary Bladder ++++++++++ 35 

Systemic Lesions

Multiple Organs + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Leukemia Mononuclear X X X X 5 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

110 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

Number of Days on Study

4444455555555555666666666 

4446901123445669002344446 

1452210772279484235835680 

Carcass ID Number

3333333333333333333333333 

3204543102404121330311430 

4614059160279237219360102 

Alimentary System

Esophagus

Periesophageal tissue, lipoma

Intestine large, colon

Intestine large, rectum

Intestine large, cecum

Intestine small, duodenum

Intestine small, jejunum

Intestine small, ileum

Liver

Mesentery

Pancreas

Salivary glands

Stomach, forestomach

Stomach, glandular

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+ ++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Adrenal medulla +++++++++++++++++++++++++ 

Pheochromocytoma benign

Islets, pancreatic +++++++++++++++++++++++++ 

Adenoma

Parathyroid gland ++++++ M ++++++++++++++++ M + 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X XXX X XXX XXXX X

Thyroid gland +++++++++++++++++++++++++ 

C-cell, adenoma X X

General Body System

None

Genital System

Clitoral gland +++++++++++++++++++++++++ 

Adenoma X

Carcinoma X

Bilateral, adenoma

Ovary +++++++++++++++++++++++++ 

Uterus +++++++++++++++++++++++++ 

Polyp stromal X

Sarcoma stromal

Vagina + 

Squamous cell carcinoma X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 111 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

6667777777777777777777777 

Number of Days on Study 6790011133333333333333333 

6376822766666666666666666 

3333333333333333333333333 Total

Carcass ID Number 2230104300111122222334444 Tissues/ 

9163846558345924578780378 Tumors

Alimentary System

Esophagus ++++++++ 33 

Periesophageal tissue, lipoma 1 

Intestine large, colon ++++++++ 33 

Intestine large, rectum ++++++++ 33 

Intestine large, cecum ++++++++ 33 

Intestine small, duodenum ++++++++ 33 

Intestine small, jejunum ++++++++ 33 

Intestine small, ileum M +++++++ 32 

Liver ++++++++ 33 

Mesentery + 4 

Pancreas ++++++++ 33 

Salivary glands ++++++++ 33 

Stomach, forestomach ++++++++ 33 

Stomach, glandular ++++++++ 33 

Cardiovascular System

Blood vessel ++++++++ 33 

Heart ++++++++ 33 

Endocrine System

Adrenal cortex ++++++++ 33 

Adrenal medulla ++++++++ 33 

Pheochromocytoma benign X 1 

Islets, pancreatic ++++++++ 33 

Adenoma X 1 

Parathyroid gland ++++++++ 31 

Pituitary gland ++++++++ 33 

Pars distalis, adenoma X X X X X X 19 

Thyroid gland ++++++++ 33 

C-cell, adenoma 2 

General Body System

None

Genital System

Clitoral gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adenoma X X 3 

Carcinoma 1 

Bilateral, adenoma X 1 

Ovary ++++++++ 33 

Uterus ++++++++ 33 

Polyp stromal 1 

Sarcoma stromal X 1 

Vagina 1 

Squamous cell carcinoma 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

112 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

4444455555555555666666666 

Number of Days on Study 4446901123445669002344446 

1452210772279484235835680 

3333333333333333333333333 

Carcass ID Number 3204543102404121330311430 

4614059160279237219360102 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Lymph node ++++ +++++++++++ ++ ++ 

Lymph node, mandibular +++++++++++++++++++++++++ 

Lymph node, mesenteric +++++++++++++++++++++++++ 

Spleen +++++++++++++++++++++++++ 

Thymus +++++++++++++++++ M +++++++ 

Integumentary System

Mammary gland +++++++++++++++++++++++++ 

Fibroadenoma X X X X

Skin +++++++++++++++++++++++++ 

Subcutaneous tissue, sarcoma X

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Nervous System

Brain +++++++++++++++++++++++++ 

Respiratory System

Lung +++++++++++++++++++++++++ 

Nose +++++++++++++++++++++++++ 

Trachea +++++++++++++++++++++++++ 

Special Senses System

Eye + + 

Urinary System

Kidney +++++++++++++++++++++++++ 

Urinary bladder +++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 

Leukemia mononuclear X X X X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 113 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 120 mg/kg

(continued) 

6667777777777777777777777 

Number of Days on Study 6790011133333333333333333 

6376822766666666666666666 

3333333333333333333333333 Total

Carcass ID Number 2230104300111122222334444 Tissues/ 

9163846558345924578780378 Tumors

Hematopoietic System

Bone marrow ++++++++ 33 

Lymph node ++++++++ 27 

Lymph node, mandibular ++++++++ 33 

Lymph node, mesenteric ++++++++ 33 

Spleen ++++++++ 33 

Thymus +++++ M + + 31 

Integumentary System

Mammary gland ++++++++ 33 

Fibroadenoma X X X X 8 

Skin +++++++++++++++++++++++++ 50 

Subcutaneous tissue, sarcoma 1 

Musculoskeletal System

Bone ++++++++ 33 

Nervous System

Brain ++++++++ 33 

Respiratory System

Lung ++++++++ 33 

Nose ++++++++ 33 

Trachea ++++++++ 33 

Special Senses System

Eye 2 

Urinary System

Kidney ++++++++ 33 

Urinary bladder ++++++++ 33 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 50 

Leukemia mononuclear X X X X 9 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

114 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

2333334455555555566666666 

Number of Days on Study 8237991401225789911333445 

9489162175066291615034061 

3333333333333333333333333 

Carcass ID Number 6656987798557788589999768 

2976329268524663154729149 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Intestine large, rectum +++++++++++++++++++++++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Intestine small, duodenum +++++++++++++++++++++++++ 

Intestine small, jejunum +++++++++++++++++++++++++ 

Intestine small, ileum +++++++++++++++++++++++++ 

Liver +++++++++++++++++++++++++ 

Mesentery + + 

Oral mucosa

Pancreas +++++++++++++++++++++++++ 

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Stomach, glandular +++++++++++++++++++++++++ 

Tooth

Cardiovascular System

Blood vessel

Heart

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Adenoma

Adrenal medulla +++++++++++++++++++++++++ 

Pheochromocytoma benign X

Islets, pancreatic +++++++++++++++++++++++++ 

Parathyroid gland + M ++++ M +++ M + M ++++++++++ M + 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X XXX XXXXXX X X

Pars distalis, adenoma, multiple

Pars intermedia, adenoma

Thyroid gland +++++++++++++++++++++++++ 

C-cell, adenoma

C-cell, adenoma, multiple

Follicular cell, adenoma X

Follicular cell, carcinoma

General Body System

None

Genital System

Clitoral gland +++++++++++++++++++++++++ 

Carcinoma X

Ovary +++++++++++++++++++++++++ 

Uterus +++++++++++++++++++++++++ 

Polyp stromal X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 115 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

6666677777777777777777777 

Number of Days on Study 5779901123333333333333333 

3475661486666666666666666 

4333333333333333333333333 Total

Carcass ID Number 0956756695556667777888899 Tissues/ 

0093538154680570378014718 Tumors

Alimentary System

Esophagus +++++++++++++++++++++++++ 50 

Intestine large, colon +++++++++++++++++++++++++ 50 

Intestine large, rectum +++++++++++++++++++++++++ 50 

Intestine large, cecum +++++++++++++++++++++++++ 50 

Intestine small, duodenum +++++++++++++++++++++++++ 50 

Intestine small, jejunum +++++++++++++++++++++++++ 50 

Intestine small, ileum +++++++++++++++++++++++++ 50 

Liver +++++++++++++++++++++++++ 50 

Mesentery + + 4 

Oral mucosa + + 2 

Pancreas +++++++++++++++++++++++++ 50 

Salivary glands +++++++++++++++++++++++++ 50 

Stomach, forestomach +++++++++++++++++++++++++ 50 

Stomach, glandular +++++++++++++++++++++++++ 50 

Tooth + 1 

Cardiovascular System

Blood vessel

Heart

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

50 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 50 

Adenoma X 1 

Adrenal medulla +++++++++++++++++++++++++ 50 

Pheochromocytoma benign X 2 

Islets, pancreatic +++++++++++++++++++++++++ 50 

Parathyroid gland +++++++++++++++++++++++++ 45 

Pituitary gland +++++++++++++++++++++++++ 50 

Pars distalis, adenoma X X X X XXX X X XXX X X X 27 

Pars distalis, adenoma, multiple X X 2 

Pars intermedia, adenoma X 1 

Thyroid gland +++++++++++++++++++++++++ 50 

C-cell, adenoma X X X X X X X 7 

C-cell, adenoma, multiple X 1 

Follicular cell, adenoma 1 

Follicular cell, carcinoma X 1 

General Body System

None

Genital System

Clitoral gland +++++++++++++++++++++++++ 50 

Carcinoma X 2 

Ovary +++++++++++++++++++++++++ 50 

Uterus +++++++++++++++++++++++++ 50 

Polyp stromal X X 3 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

116 Sodium Xylenesulfonate, NTP TR 464 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

2333334455555555566666666 

Number of Days on Study 8237991401225789911333445 

9489162175066291615034061 

3333333333333333333333333 

Carcass ID Number 6656987798557788589999768 

2976329268524663154729149 

Hematopoietic System

Bone marrow +++++++++++++++ M +++++++++ 

Lymph node ++++ +++++ ++++ ++++++ + 

Lymph node, mandibular +++++++++++++++++++++++++ 

Lymph node, mesenteric ++++++++++++++++++++ M ++++ 

Spleen +++++++++++++++++++++++++ 

Thymus +++++++++++++++++++++++++ 

Integumentary System

Mammary gland +++++++++++++++++++++++++ 

Carcinoma X

Fibroadenoma X X

Fibroadenoma, multiple

Skin +++++++++++++++++++++++++ 

Musculoskeletal System

Bone +++++++++++++++ M +++++++++ 

Nervous System

Brain +++++++++++++++++++++++++ 

Respiratory System

Lung +++++++++++++++++++++++++ 

Nose +++++++++++++++++++++++++ 

Trachea +++++++++++++++++++++++++ 

Special Senses System

Eye

Zymbal’s gland + 

Carcinoma X

Urinary System

Kidney +++++++++++++++++++++++++ 

Urinary bladder +++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 

Leukemia mononuclear X X X X

Mesothelioma benign
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 117 

TABLE B2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate: 240 mg/kg

(continued) 

6666677777777777777777777 

Number of Days on Study 5779901123333333333333333 

3475661486666666666666666 

4333333333333333333333333 Total

Carcass ID Number 0956756695556667777888899 Tissues/ 

0093538154680570378014718 Tumors

Hematopoietic System

Bone marrow + M +++++++++++++++++++++++ 48 

Lymph node ++ ++++++++++ +++++++++++ 43 

Lymph node, mandibular +++++++++++++++++++++++++ 50 

Lymph node, mesenteric ++++++++++++++++++ M ++++++ 48 

Spleen +++++++++++++++++++++++++ 50 

Thymus ++++ M ++++++++++++++++++++ 49 

Integumentary System

Mammary gland +++++++++++++++++++++++++ 50 

Carcinoma 1 

Fibroadenoma X X X X 6 

Fibroadenoma, multiple X X X 3 

Skin +++++++++++++++++++++++++ 50 

Musculoskeletal System

Bone + M +++++++++++++++++++++++ 48 

Nervous System

Brain +++++++++++++++++++++++++ 50 

Respiratory System

Lung +++++++++++++++++++++++++ 50 

Nose +++++++++++++++++++++++++ 50 

Trachea +++++++++++++++++++++++++ 50 

Special Senses System

Eye + + + + + 5 

Zymbal’s gland 1 

Carcinoma 1 

Urinary System

Kidney +++++++++++++++++++++++++ 50 

Urinary bladder +++++++++++++++++++++++++ 50 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 50 

Leukemia mononuclear X X X X X X 10 

Mesothelioma benign X 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

118 Sodium Xylenesulfonate, NTP TR 464 

TABLE B3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Clitoral Gland: Adenoma

Overall ratea

Adjusted rateb

Terminal ratec

First incidence (days) 

Life table testd

Logistic regression testd

Cochran-Armitage testd

Fisher exact testd

1/49 (2%) 

4.5% 

1/22 (5%) 

736 (T) 

P ̆0.388N

P ̆0.340N

P ̆0.307N

3/50 (6%) 

12.7% 

1/16 (6%) 

622 

P ̆0.220 

P ̆0.263 

P ̆0.316 

4/50 (8%) 

18.6% 

2/17 (12%) 

602 

P ̆0.124 

P ̆0.146 

P ̆0.187 

0/50 (0%) 

0.0% 

0/16 (0%) 

—e

P ̆0.564N

P ̆0.564N

P ̆0.495N

Clitoral Gland: Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

1/49 (2%) 

4.5% 

1/22 (5%) 

736 (T) 

P ̆0.364 

P ̆0.420 

P ̆0.445 

3/50 (6%) 

12.7% 

1/16 (6%) 

622 

P ̆0.220 

P ̆0.263 

P ̆0.316 

5/50 (10%) 

20.4% 

2/17 (12%) 

492 

P ̆0.075 

P ̆0.100 

P ̆0.107 

2/50 (4%) 

9.1% 

1/16 (6%) 

611 

P ̆0.420 

P ̆0.460 

P ̆0.508 

Mammary Gland: Fibroadenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

16/50 (32%) 

56.3% 

10/22 (45%) 

490 

4/35 (11%) f 8/33 (24%) f 9/50 (18%) 

41.8% 

5/16 (31%) 

515 

P ̆0.285N

P ̆0.151N

P ̆0.083N

Mammary Gland: Fibroadenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

16/50 (32%) 

56.3% 

10/22 (45%) 

490 

5/35 (14%) f 8/33 (24%) f 10/50 (20%) 

43.1% 

5/16 (31%) 

441 

P ̆0.373N

P ̆0.202N

P ̆0.127N

Pituitary Gland (Pars Distalis): Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

34/50 (68%) 

84.5% 

16/22 (73%) 

420 

17/35 (49%)f 19/33 (58%)f 29/50 (58%) 

81.4% 

10/16 (63%) 

396 

P ̆0.429 

P ̆0.243N

P ̆0.204N
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 119 

TABLE B3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

(continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Pituitary Gland (Pars Distalis): Adenoma or Carcinoma

Overall rate 35/50 (70%) 

Adjusted rate 84.8% 

Terminal rate 16/22 (73%) 

First incidence (days) 420 

Life table test

Logistic regression test

Fisher exact test

17/35 (49%)f 19/33 (58%)f 29/50 (58%) 

81.4% 

10/16 (63%) 

396 

P ̆0.486 

P ̆0.182N

P ̆0.149N

Thyroid Gland (C-cell): Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

10/50 (20%) 

39.3% 

7/22 (32%) 

693 

1/35 (3%) f 2/33 (6%) f 8/50 (16%) 

40.6% 

5/16 (31%) 

674 

P ̆0.529 

P ̆0.601 

P ̆0.398N

Thyroid Gland (Follicular Cell): Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

1/50 (2%) 

4.5% 

1/22 (5%) 

736 (T) 

2/35 (6%) f 0/33 (0%) f 1/50 (2%) 

2.7% 

0/16 (0%) 

572 

P ̆0.722 

P ̆0.756 

P ̆0.753N

Thyroid Gland (Follicular Cell): Adenoma or Carcinoma

Overall rate 1/50 (2%) 

Adjusted rate 4.5% 

Terminal rate 1/22 (5%) 

First incidence (days) 736 (T) 

Life table test

Logistic regression test

Fisher exact test

2/35 (6%) f 0/33 (0%) f 2/50 (4%) 

6.9% 

0/16 (0%) 

572 

P ̆0.430 

P ̆0.483 

P ̆0.500 

Uterus: Stromal Polyp

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

2/50 (4%) 

6.5% 

1/22 (5%) 

408 

1/35 (3%) f 1/33 (3%) f 3/50 (6%) 

14.4% 

2/16 (13%) 

391 

P ̆0.401 

P ̆0.524 

P ̆0.500 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

120 Sodium Xylenesulfonate, NTP TR 464 

TABLE B3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

(continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Uterus: Stromal Polyp or Stromal Sarcoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

2/50 (4%) 

6.5% 

1/22 (5%) 

408 

2/35 (6%) f 2/33 (6%) f 3/50 (6%) 

14.4% 

2/16 (13%) 

391 

P ̆0.401 

P ̆0.524 

P ̆0.500 

All Organs: Mononuclear Cell Leukemia

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

17/50 (34%) 

53.3% 

8/22 (36%) 

485 

5/35 (14%) f 9/33 (27%) f 10/50 (20%) 

42.3% 

5/16 (31%) 

520 

P ̆0.301N

P ̆0.136N

P ̆0.088N

All Organs: Benign Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

40/50 (80%) 

95.1% 

20/22 (91%) 

408 

23/50 (46%)f 26/50 (52%)f 33/50 (66%) 

86.1% 

11/16 (69%) 

391 

P ̆0.484 

P ̆0.141N

P ̆0.088N

All Organs: Malignant Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

23/50 (46%) 

61.2% 

9/22 (41%) 

303 

8/50 (16%) f 11/50 (22%)f 15/50 (30%) 

53.5% 

6/16 (38%) 

338 

P ̆0.312N

P ̆0.077N

P ̆0.074N
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

121 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE B3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Dermal Study of Sodium Xylenesulfonate

(continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

All Organs: Benign or Malignant Neoplasms

Overall rate 45/50 (90%) 25/50 (50%)f 27/50 (54%)f 41/50 (82%) 

Adjusted rate 95.7% 95.3% 

Terminal rate 20/22 (91%) 14/16 (88%) 

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

303 338 

P ̆0.272 

P ̆0.273N

P ̆0.194N

(T)Terminal sacrifice a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for clitoral gland, mammary

gland, 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 dosed group incidence are the P values corresponding to pairwise

comparisons between the controls and that dosed 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 lower incidence in a dose group is indicated by N. e Not applicable; no neoplasms in animal group f	 Tissues (except skin) were examined microscopically only in those animals dying prior to terminal sacrifice; thus statistical comparisons with the controls are

not applicable. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

122 Sodium Xylenesulfonate, NTP TR 464 

TABLE B4 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Accidental death 1 

Moribund 14 10 15 12 

Natural deaths 13 24 18 22 

Survivors

Died last week of the study 1 

Terminal sacrifice 22 15 17 16 

Animals examined microscopically 50 50 50 50 

Alimentary System

Intestine large, colon (50) (35) (33) (50) 

Parasite metazoan 2 (4%) 1 (3%) 3 (6%) 

Intestine large, rectum (50) (35) (33) (50) 

Parasite metazoan 4 (8%) 3 (9%) 1 (3%) 3 (6%) 

Intestine large, cecum (50) (35) (33) (50) 

Inflammation, chronic active 1 (3%) 1 (3%) 1 (2%) 

Ulcer 1 (3%) 

Intestine small, duodenum (50) (35) (33) (50) 

Inflammation, chronic active 1 (3%) 

Ulcer 1 (2%) 1 (3%) 

Liver (50) (35) (33) (50) 

Angiectasis 1 (2%) 

Basophilic focus 30 (60%) 12 (34%) 11 (33%) 26 (52%) 

Clear cell focus 1 (2%) 1 (2%) 

Hematopoietic cell proliferation 1 (2%) 

Hepatodiaphragmatic nodule 10 (20%) 6 (17%) 11 (33%) 3 (6%) 

Inflammation, chronic 21 (42%) 17 (49%) 19 (58%) 24 (48%) 

Mixed cell focus 1 (2%) 

Necrosis 3 (6%) 1 (3%) 3 (9%) 3 (6%) 

Pigmentation, hemosiderin 3 (6%) 1 (3%) 

Pigmentation, melanin 1 (2%) 

Bile duct, hyperplasia 28 (56%) 13 (37%) 17 (52%) 21 (42%) 

Central vein, thrombosis 1 (2%) 

Hepatocyte, hyperplasia 2 (4%) 1 (3%) 1 (2%) 

Hepatocyte, vacuolization cytoplasmic 15 (30%) 4 (12%) 4 (8%) 

Hepatocyte, centrilobular, degeneration, 

fatty 4 (8%) 1 (3%) 3 (9%) 2 (4%) 

Portal vein, thrombosis 1 (2%) 

Mesentery (2) (4) (4) (4) 

Mineralization 1 (50%) 2 (50%) 1 (25%) 

Artery, inflammation, chronic active 1 (25%) 

Fat, inflammation, chronic active 1 (50%) 2 (50%) 1 (25%) 2 (50%) 

Fat, necrosis 1 (50%) 2 (50%) 

Oral mucosa (1) (2) 

Gingival, inflammation, chronic active 2 (100%) 

a Number of animals examined microscopically at the site and the number of animals with lesion
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 123 

TABLE B4 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Alimentary System (continued) 

Pancreas (50) (35) (33) (50) 

Cyst 1 (3%) 

Hypertrophy, focal 1 (2%) 

Inflammation, chronic active 1 (3%) 1 (3%) 

Acinus, atrophy 12 (24%) 3 (9%) 5 (15%) 8 (16%) 

Artery, inflammation, chronic active 1 (2%) 1 (2%) 

Stomach, forestomach (50) (35) (33) (50) 

Inflammation, chronic active 2 (4%) 4 (11%) 3 (9%) 4 (8%) 

Ulcer 2 (4%) 2 (6%) 3 (9%) 3 (6%) 

Epithelium, hyperplasia 2 (4%) 4 (11%) 3 (9%) 5 (10%) 

Stomach, glandular (50) (35) (33) (50) 

Inflammation, chronic active 1 (3%) 1 (3%) 

Mineralization 1 (2%) 1 (3%) 

Necrosis 1 (2%) 1 (3%) 

Ulcer 1 (3%) 

Epithelium, erosion 1 (2%) 1 (3%) 

Epithelium, hemorrhage 1 (2%) 

Epithelium, necrosis 1 (2%) 

Tooth (1) (1) 

Inflammation 1 (100%) 

Peridontal tissue, inflammation, chronic

active 1 (100%) 

Cardiovascular System

Blood vessel (50) (35) (33) (50) 

Mineralization 1 (2%) 1 (3%) 

Heart (50) (35) (33) (50) 

Cardiomyopathy, chronic 16 (32%) 10 (29%) 15 (45%) 23 (46%) 

Inflammation, chronic active 1 (3%) 

Mineralization 1 (2%) 1 (3%) 

Atrium, thrombosis 4 (8%) 1 (3%) 2 (6%) 2 (4%) 

Endocrine System

Adrenal cortex (50) (35) (33) (50) 

Accessory adrenal cortical nodule 2 (4%) 1 (3%) 1 (2%) 

Atrophy 1 (2%) 

Degeneration, fatty 24 (48%) 12 (34%) 7 (21%) 14 (28%) 

Hyperplasia 17 (34%) 10 (29%) 6 (18%) 17 (34%) 

Hypertrophy 1 (2%) 1 (3%) 1 (2%) 

Karyomegaly 1 (3%) 1 (2%) 

Necrosis 1 (2%) 

Pigmentation, hemosiderin 1 (2%) 

Pigmentation, lipofuscin 3 (6%) 2 (6%) 4 (12%) 7 (14%) 

Adrenal medulla (50) (35) (33) (50) 

Hyperplasia 9 (18%) 3 (9%) 1 (3%) 5 (10%) 

Islets, pancreatic (50) (35) (33) (50) 

Hyperplasia 1 (2%) 1 (2%) 

Parathyroid gland (48) (32) (31) (45) 

Hyperplasia 1 (2%) 1 (3%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

124 Sodium Xylenesulfonate, NTP TR 464 

TABLE B4 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Endocrine System (continued) 

Pituitary gland (50) (35) (33) (50) 

Angiectasis 36 (72%) 21 (60%) 19 (58%) 31 (62%) 

Mineralization 1 (2%) 1 (3%) 

Pigmentation, hematoidin 1 (2%) 1 (3%) 1 (2%) 

Pars distalis, atypia cellular 1 (2%) 

Pars distalis, cyst 21 (42%) 11 (31%) 15 (45%) 29 (58%) 

Pars distalis, hyperplasia 9 (18%) 11 (31%) 7 (21%) 17 (34%) 

Pars distalis, pigmentation, hemosiderin 32 (64%) 20 (57%) 20 (61%) 31 (62%) 

Pars distalis, thrombosis 1 (3%) 

Pars intermedia, cyst 2 (6%) 2 (4%) 

Thyroid gland (50) (35) (33) (50) 

Ultimobranchial cyst 1 (2%) 

C-cell, hyperplasia 7 (14%) 5 (14%) 5 (15%) 8 (16%) 

Follicle, cyst 1 (3%) 1 (2%) 

General Body System

None

Genital System

Clitoral gland (49) (50) (50) (50) 

Hyperplasia 2 (4%) 2 (4%) 1 (2%) 10 (20%) 

Inflammation, chronic active 6 (12%) 4 (8%) 4 (8%) 7 (14%) 

Duct, cyst 1 (2%) 2 (4%) 2 (4%) 

Duct, hyperplasia 2 (4%) 

Ovary (50) (35) (33) (50) 

Cyst 3 (6%) 1 (3%) 2 (6%) 4 (8%) 

Inflammation, chronic active 1 (3%) 

Interstitial cell, hyperplasia 2 (6%) 

Uterus (50) (35) (33) (50) 

Cyst 2 (4%) 1 (3%) 

Vagina (1) (1) (1) 

Inflammation, suppurative 1 (100%) 

Hematopoietic System

Bone marrow (50) (35) (33) (48) 

Hyperplasia 19 (38%) 4 (11%) 12 (36%) 10 (21%) 

Inflammation, granulomatous 1 (2%) 

Lymph node (45) (32) (27) (43) 

Lumbar, hyperplasia, lymphoid 1 (2%) 

Mediastinal, hyperplasia, plasma cell 1 (2%) 

Mediastinal, pigmentation, hemosiderin 44 (98%) 31 (97%) 27 (100%) 40 (93%) 

Pancreatic, pigmentation, hemosiderin 1 (2%) 

Renal, pigmentation, hemosiderin 1 (2%) 

Lymph node, mandibular (50) (35) (33) (50) 

Pigmentation, hemosiderin 2 (4%) 5 (14%) 6 (18%) 4 (8%) 

Lymph node, mesenteric (49) (35) (33) (48) 

Ectasia 1 (2%) 

Inflammation, suppurative 1 (2%) 

Pigmentation, hemosiderin 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 125 

TABLE B4 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Hematopoietic System (continued) 

Spleen (50) (35) (33) (50) 

Accessory spleen 2 (4%) 

Fibrosis 3 (6%) 

Hematopoietic cell proliferation 2 (4%) 4 (11%) 3 (9%) 1 (2%) 

Necrosis 1 (3%) 

Pigmentation, hemosiderin 4 (8%) 5 (14%) 9 (27%) 11 (22%) 

Capsule, fibrosis 1 (3%) 

Capsule, inflammation, chronic active 1 (3%) 

Lymphoid follicle, depletion cellular 1 (3%) 

Thymus (49) (35) (31) (49) 

Cyst 1 (2%) 1 (3%) 

Integumentary System

Mammary gland (50) (35) (33) (50) 

Hyperplasia, cystic 42 (84%) 23 (66%) 27 (82%) 42 (84%) 

Duct, cyst 1 (2%) 1 (2%) 

Skin (50) (50) (50) (50) 

Hyperkeratosis 1 (2%) 

Hyperplasia, basal cell 1 (2%) 

Inflammation, chronic active 1 (2%) 

Epidermis, hyperplasia 1 (2%) 

Epidermis, ulcer 1 (2%) 

Epidermis, skin, site of application, exudate 2 (4%) 

Epidermis, skin, site of application, 

hyperplasia 1 (2%) 4 (8%) 5 (10%) 

Epidermis, skin, site of application, 

inflammation, chronic active 1 (2%) 

Epidermis, skin, site of application, ulcer 1 (2%) 2 (4%) 2 (4%) 

Sebaceous gland, skin, site of application, 

hyperplasia 2 (4%) 2 (4%) 2 (4%) 

Skin, site of application, inflammation, 

chronic active 1 (2%) 2 (4%) 2 (4%) 

Musculoskeletal System

Bone (50) (35) (33) (48) 

Fibrous osteodystrophy 1 (2%) 1 (3%) 

Osteopetrosis 1 (2%) 1 (3%) 

Femur, osteosclerosis 1 (3%) 1 (2%) 

Femur, tibia, osteosclerosis 1 (2%) 

Joint, arthrosis 1 (3%) 

Joint, tarsal, arthrosis 1 (3%) 

Tibia, fracture 1 (3%) 

Skeletal muscle (1) 

Inflammation, chronic active 1 (100%) 

Nervous System

Brain (50) (35) (33) (50) 

Hemorrhage, acute 1 (2%) 1 (2%) 

Hydrocephalus 8 (16%) 5 (14%) 3 (9%) 7 (14%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

126 Sodium Xylenesulfonate, NTP TR 464 

TABLE B4 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 60 mg/kg 120 mg/kg 240 mg/kg

Respiratory System

Lung (50) (35) (33) (50) 

Congestion, acute 13 (26%) 14 (40%) 8 (24%) 16 (32%) 

Foreign body 1 (3%) 

Inflammation, chronic active 3 (6%) 2 (6%) 2 (6%) 4 (8%) 

Inflammation, granulomatous 2 (4%) 

Inflammation, suppurative 1 (2%) 

Mineralization 1 (2%) 1 (3%) 

Pigmentation, hemosiderin 1 (3%) 

Alveolar epithelium, hyperplasia 1 (3%) 4 (8%) 

Alveolus, hemorrhage, acute 1 (3%) 

Alveolus, infiltration cellular, histiocyte 32 (64%) 23 (66%) 20 (61%) 39 (78%) 

Interstitium, inflammation, chronic active 9 (18%) 4 (11%) 2 (6%) 5 (10%) 

Nose (50) (35) (33) (50) 

Inflammation, chronic active 4 (8%) 2 (6%) 6 (12%) 

Nasolacrimal duct, inflammation, chronic

active 1 (2%) 

Nasolacrimal duct, inflammation, suppurative 10 (20%) 3 (9%) 1 (3%) 6 (12%) 

Special Senses System

Eye (3) (2) (2) (5) 

Cornea, inflammation, suppurative 1 (33%) 

Lens, cataract 1 (33%) 2 (100%) 2 (100%) 3 (60%) 

Retina, degeneration 1 (33%) 1 (50%) 2 (100%) 4 (80%) 

Retina, hemorrhage 1 (20%) 

Urinary System

Kidney (50) (35) (33) (50) 

Infarct 1 (3%) 

Mineralization 10 (20%) 22 (63%) 7 (21%) 11 (22%) 

Nephropathy, chronic 44 (88%) 25 (71%) 27 (82%) 37 (74%) 

Pelvis, transitional epithelium, hyperplasia 1 (3%) 

Renal tubule, pigmentation, hemosiderin 1 (3%) 
 

 

 

 

 

 

 

 

 

 

 

 

127 

APPENDIX C
 

SUMMARY OF LESIONS IN MALE MICE
 

IN THE 2-YEAR DERMAL STUDY
 

OF SODIUM XYLENESULFONATE
 

TABLE C1 Summary of the Incidence of Neoplasms in Male Mice
 

in the 2-Year Dermal Study of Sodium Xylenesulfonate
 ...................... 129 
 #

TABLE C2 Individual Animal Tumor Pathology of Male Mice
 

...................... 132 
 #

TABLE C3 Statistical Analysis of Primary Neoplasms in Male Mice
 

...................... 152 
 #

in the 2-Year Dermal Study of Sodium Xylenesulfonate
 

in the 2-Year Dermal Study of Sodium Xylenesulfonate
 

TABLE C4	# Historical Incidence of Hepatocellular Neoplasms in Control Male B6C3F1 Mice . . . . . 156 
 #

TABLE C5	# Summary of the Incidence of Nonneoplastic Lesions in Male Mice
 

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 157 
 #
128 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 129 

TABLE C1 

Summary of the Incidence of Neoplasms in Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Moribund 7 3 8 6 

Natural deaths 11 10 3 9 

Survivors

Terminal sacrifice 32 37 39 35 

Animals examined microscopically 50 50 50 50 

Alimentary System

Intestine small, jejunum (50) (12) (11) (50) 

Liver (50) (50) (50) (50) 

Hemangioma 2 (4%) 1 (2%) 

Hemangiosarcoma 2 (4%) 2 (4%) 1 (2%) 

Hemangiosarcoma, multiple 3 (6%) 3 (6%) 2 (4%) 1 (2%) 

Hemangiosarcoma, metastatic, spleen 1 (2%) 1 (2%) 1 (2%) 3 (6%) 

Hepatoblastoma 4 (8%) 2 (4%) 

Hepatocellular carcinoma 21 (42%) 17 (34%) 12 (24%) 15 (30%) 

Hepatocellular carcinoma, multiple 14 (28%) 14 (28%) 10 (20%) 11 (22%) 

Hepatocellular adenoma 15 (30%) 21 (42%) 14 (28%) 13 (26%) 

Hepatocellular adenoma, multiple 22 (44%) 11 (22%) 7 (14%) 16 (32%) 

Hepatocholangiocarcinoma 1 (2%) 

Histiocytic sarcoma 1 (2%) 

Osteosarcoma, metastatic, uncertain primary

site 1 (2%) 

Mesentery (5) (4) 

Hemangiosarcoma, metastatic, spleen 1 (20%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (20%) 

Liposarcoma, multiple 1 (25%) 

Pancreas (50) (13) (11) (50) 

Liposarcoma, metastatic, mesentery 1 (2%) 

Salivary glands (50) (13) (11) (50) 

Stomach, forestomach (50) (13) (11) (50) 

Squamous cell papilloma 1 (2%) 1 (2%) 

Stomach, glandular (50) (13) (11) (50) 

Cardiovascular System

Heart (50) (13) (11) (50) 

Alveolar/bronchiolar carcinoma, metastatic, 

lung 1 (8%) 

Hemangiosarcoma, metastatic, spleen 1 (2%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (2%) 

Endocrine System

Adrenal cortex (50) (12) (11) (50) 

Adrenal medulla (50) (12) (11) (50) 

Pheochromocytoma benign 2 (4%) 

Pituitary gland (49) (12) (10) (50) 

Pars distalis, adenoma 1 (2%) 

Thyroid gland (50) (13) (11) (50) 

Follicular cell, adenoma 2 (4%) 1 (9%) 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

130 Sodium Xylenesulfonate, NTP TR 464 

TABLE C1
#

Summary of the Incidence of Neoplasms in Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

General Body System

None

Genital System

Epididymis (50) (13) (11) (50) 

Liposarcoma, metastatic, mesentery 1 (2%) 

Prostate (50) (13) (11) (50) 

Liposarcoma, metastatic, mesentery 1 (2%) 

Seminal vesicle (50) (13) (11) (50) 

Hematopoietic System

Bone marrow (50) (13) (11) (50) 

Hemangiosarcoma 1 (9%) 

Hemangiosarcoma, metastatic, spleen 1 (2%) 1 (9%) 

Lymph node (3) (2) (2) (3) 

Mediastinal, alveolar/bronchiolar carcinoma, 

metastatic, lung 1 (50%) 

Mediastinal, hemangiosarcoma, metastatic, 

liver 1 (33%) 

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver 1 (33%) 

Mediastinal, histiocytic sarcoma 1 (50%) 

Lymph node, mandibular (43) (7) (8) (42) 

Lymph node, mesenteric (50) (9) (10) (47) 

Hemangioma 1 (2%) 1 (2%) 

Hemangiosarcoma, metastatic, spleen 1 (2%) 

Spleen (50) (13) (12) (49) 

Hemangiosarcoma 2 (4%) 1 (8%) 1 (8%) 3 (6%) 

Hemangiosarcoma, metastatic, bone marrow 1 (8%) 

Liposarcoma, metastatic, mesentery 1 (2%) 

Thymus (37) (11) (6) (34) 

Alveolar/bronchiolar carcinoma, metastatic, 

lung 1 (9%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (3%) 

Histiocytic sarcoma 1 (9%) 

Integumentary System

Skin (50) (50) (50) (50) 

Subcutaneous tissue, fibrosarcoma 1 (2%) 

Subcutaneous tissue, skin, site of

application, hemangioma 1 (2%) 

Subcutaneous tissue, skin, site of

application, hemangiosarcoma 1 (2%) 

Subcutaneous tissue, skin, site of

application, histiocytic sarcoma 2 (4%) 

Subcutaneous tissue, skin, site of

application, lymphoma malignant 1 (2%) 

Musculoskeletal System

Skeletal muscle (3) 

Hepatocellular carcinoma, metastatic, liver 1 (33%) 

Liposarcoma, metastatic, mesentery 1 (33%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 131 

TABLE C1
#

Summary of the Incidence of Neoplasms in Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Nervous System

Brain (50) (13) (11) (50) 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar adenoma, multiple

Alveolar/bronchiolar carcinoma

Hemangiosarcoma, metastatic, spleen

Hepatoblastoma, metastatic, liver

Hepatocellular carcinoma, metastatic, liver

Hepatocholangiocarcinoma, metastatic, liver

Histiocytic sarcoma

Mediastinum, alveolar/bronchiolar carcinoma, 

metastatic, lung

(50) 

 12 (24%) 

 1 (2%) 

 2 (4%) 

 12 (24%) 

 1 (2%) 

(16) 

 4 (25%) 

 1 (6%) 

 5 (31%) 

 1 (6%) 

 1 (6%) 

(15) 

 2 (13%) 

 1 (7%) 

 7 (47%) 

(50) 

 7 (14%) 

 2 (4%) 

 1 (2%) 

 1 (2%) 

 2 (4%) 

 11 (22%) 

Special Senses System

Harderian gland

Adenoma

Carcinoma

 (2)

 1 (50%) 

(1)

 1 (100%)

 (3) 

 1 (33%) 

 1 (33%) 

Urinary System

Kidney

Hepatocellular carcinoma, metastatic, liver

Renal tubule, adenoma

Renal tubule, carcinoma

(50) (13) 

 1 (8%) 

(11) (50) 

 1 (2%) 

 1 (2%) 

 1 (2%) 

Systemic Lesions

Multiple organsb

Histiocytic sarcoma

Lymphoma malignant

(50) 

 5 (10%) 

(50) 

 2 (4%) 

(50) 

 4 (8%) 

(50) 

 6 (12%) 

Neoplasm Summary

Total animals with primary neoplasmsc

Total primary neoplasms

Total animals with benign neoplasms

Total benign neoplasms

Total animals with malignant neoplasms

Total malignant neoplasms

Total animals with metastatic neoplasms

Total metastatic neoplasms

Total animals with malignant neoplasms

of uncertain primary site

49 

108 

43 

58 

39 

50 

15 

21 

41 

78 

33 

36 

33 

42 

6 

11 

36 

60 

24 

26 

28 

34 

10 

11 

1 

47 

90 

37 

46 

33 

44 

16 

26 

a

b

c

Number of animals examined microscopically at the site and the number of animals with neoplasm

Number of animals with any tissue examined microscopically

Primary neoplasms: all neoplasms except metastatic neoplasms
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

132 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 

Vehicle Control

4555666666666667777777777 

Number of Days on Study 9677445788899990003333333 

4455671406701172563333333 

0000000000000000000000000 

Carcass ID Number 4214100513100423201222333 

1700654036192217574028034 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Gallbladder ++++++++++++ I ++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Intestine large, rectum +++++++++++++++++++ M +++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Intestine small, duodenum +++++++++++++++++++++++++ 

Intestine small, jejunum +++++++++++++++++++++++++ 

Intestine small, ileum +++++++++++++++++++++++++ 

Liver +++++++++++++++++++++++++ 

Hemangioma X

Hemangiosarcoma X

Hemangiosarcoma, multiple X X

Hemangiosarcoma, metastatic, spleen X

Hepatocellular carcinoma X X XXXX X X X X X X

Hepatocellular carcinoma, multiple X X X X X

Hepatocellular adenoma XXXXX X X X X X

Hepatocellular adenoma, multiple X X X X X X X

Hepatocholangiocarcinoma X

Mesentery + + + + 

Hemangiosarcoma, metastatic, spleen X

Hepatocholangiocarcinoma, metastatic, 

liver X

Pancreas +++++++++++++++++++++++++ 

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Squamous cell papilloma X

Stomach, glandular +++++++++++++++++++++++++ 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Hepatocholangiocarcinoma, metastatic, liver X

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Adrenal medulla +++++++++++++++++++++++++ 

Islets, pancreatic +++++++++++++++++++++++++ 

Parathyroid gland +++ M ++++++++ M ++++++++++++ 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X

Thyroid gland +++++++++++++++++++++++++ 

Follicular cell, adenoma X

General Body System

None

+: Tissue examined microscopically M: Missing tissue X: Lesion present

A: Autolysis precludes examination I: Insufficient tissue Blank: Not examined
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 133 

TABLE C2
#

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

3334444444455555555555555 

0000000000000000000000000 Total

Carcass ID Number 3340012233400111122344444 Tissues/ 

5931853912536278946846789 Tumors

Alimentary System

Esophagus +++++++++++++++++++++++++ 50 

Gallbladder +++++++++++++++++++++++++ 49 

Intestine large, colon +++++++++++++++++++++++++ 50 

Intestine large, rectum +++++++++++++++++++++++++ 49 

Intestine large, cecum +++++++++++++++++++++++++ 50 

Intestine small, duodenum +++++++++++++++++++++++++ 50 

Intestine small, jejunum +++++++++++++++++++++++++ 50 

Intestine small, ileum +++++++++++++++++++++++++ 50 

Liver +++++++++++++++++++++++++ 50 

Hemangioma X 2 

Hemangiosarcoma X 2 

Hemangiosarcoma, multiple X 3 

Hemangiosarcoma, metastatic, spleen 1 

Hepatocellular carcinoma X X X X X X X X X 21 

Hepatocellular carcinoma, multiple XXXX X X X X X 14 

Hepatocellular adenoma X X X X X 15 

Hepatocellular adenoma, multiple X X X XXXX X X X X X X X X 22 

Hepatocholangiocarcinoma 1 

Mesentery + 5 

Hemangiosarcoma, metastatic, spleen 1 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Pancreas +++++++++++++++++++++++++ 50 

Salivary glands +++++++++++++++++++++++++ 50 

Stomach, forestomach +++++++++++++++++++++++++ 50 

Squamous cell papilloma 1 

Stomach, glandular +++++++++++++++++++++++++ 50 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 50 

Heart +++++++++++++++++++++++++ 50 

Hepatocholangiocarcinoma, metastatic, liver 1 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 50 

Adrenal medulla +++++++++++++++++++++++++ 50 

Islets, pancreatic +++++++++++++++++++++++++ 50 

Parathyroid gland ++++++++ M +++++++++++++++ M 46 

Pituitary gland +++++++++++++++++++++++ M + 49 

Pars distalis, adenoma 1 

Thyroid gland +++++++++++++++++++++++++ 50 

Follicular cell, adenoma X 2 

General Body System

None
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

134 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2
#

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

4555666666666667777777777 

Number of Days on Study 9677445788899990003333333 

4455671406701172563333333 

0000000000000000000000000 

Carcass ID Number 4214100513100423201222333 

1700654036192217574028034 

Genital System

Epididymis

Preputial gland

Prostate

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Seminal vesicle +++++++++++++++++++++++++ 

Testes +++++++++++++++++++++++++ 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Hemangiosarcoma, metastatic, spleen

Lymph node

Mediastinal, hemangiosarcoma, 

metastatic, liver

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver

Lymph node, mandibular

Lymph node, mesenteric

Hemangioma

Spleen

Hemangiosarcoma

Thymus

Hepatocholangiocarcinoma, metastatic, liver

X

+ + 

X

+++ M M + + M + + M ++++++++++++ M + 

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

X

+++ M +++++ M ++++ MMMM ++++ M ++ 

X

Integumentary System

Mammary gland

Skin

+ MMMM + M M + M M + MMM + MMM + M M + M M

+++++++++++++++++++++++++ 

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Nervous System

Brain +++++++++++++++++++++++++ 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar adenoma, multiple

Alveolar/bronchiolar carcinoma

Hepatocellular carcinoma, metastatic, liver

Hepatocholangiocarcinoma, metastatic, liver

Nose

+++++++++++++++++++++++++ 

X X X

X X

X X X X X

X

+++++++++++++++++++++++++ 

Trachea +++++++++++++++++++++++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 135 

TABLE C2
#

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

3334444444455555555555555 

0000000000000000000000000 Total

Carcass ID Number 3340012233400111122344444 

5931853912536278946846789 

Tissues/ 

Tumors

Genital System

Epididymis

Preputial gland

Prostate

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

50 

50 

Seminal vesicle +++++++++++++++++++++++++ 50 

Testes +++++++++++++++++++++++++ 50 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 50 

Hemangiosarcoma, metastatic, spleen

Lymph node

Mediastinal, hemangiosarcoma, 

metastatic, liver

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver

Lymph node, mandibular

Lymph node, mesenteric

Hemangioma

Spleen

Hemangiosarcoma

Thymus

Hepatocholangiocarcinoma, metastatic, liver

+ 

X

+++++++++ M M ++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

+++ M M ++++ M M +++ M ++++++++ M + 

1 

3 

1 

1 

43 

50 

1 

50 

2 

37 

1 

Integumentary System

Mammary gland

Skin

+ + M + M + + MMM + + MMM + + M + M M + MMM

+++++++++++++++++++++++++ 

18 

50 

Musculoskeletal System

Bone +++++++++++++++++++++++++ 50 

Nervous System

Brain +++++++++++++++++++++++++ 50 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar adenoma, multiple

Alveolar/bronchiolar carcinoma

Hepatocellular carcinoma, metastatic, liver

Hepatocholangiocarcinoma, metastatic, liver

Nose

+++++++++++++++++++++++++ 

X X X X X X X X X

X

X X X X X X X

+++++++++++++++++++++++++ 

50 

12 

1 

2 

12 

1 

50 

Trachea +++++++++++++++++++++++++ 50 
 

 

 

 

 

 

 

 

 

 

136 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2
#

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

4555666666666667777777777 

Number of Days on Study 9677445788899990003333333 

4455671406701172563333333 

0000000000000000000000000 

Carcass ID Number 4214100513100423201222333 

1700654036192217574028034 

Special Senses System

Harderian gland + 

Adenoma X

Urinary System

Kidney +++++++++++++++++++++++++ 

Urinary bladder +++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 

Lymphoma malignant X X X X X
 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 137 

TABLE C2
#

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

3334444444455555555555555 

0000000000000000000000000 Total

Carcass ID Number 3340012233400111122344444 Tissues/ 

5931853912536278946846789 Tumors

Special Senses System

Harderian gland + 2 

Adenoma 1 

Urinary System

Kidney +++++++++++++++++++++++++ 50 

Urinary bladder +++++++++++++++++++++++++ 50 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 50 

Lymphoma malignant 5 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

138 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

3556666666677777777777777 

Number of Days on Study 2481235578800333333333333 

2080098940726333333334444 

0010000000000000000000000 

Carcass ID Number 6607887957659588889995566 

0708801813691612570353413 

Alimentary System

Esophagus +++++++++++++ 

Gallbladder ++++++++ M ++++ 

Intestine large, colon +++++++++++++ 

Intestine large, rectum +++++++++++++ 

Intestine large, cecum +++++++++++++ 

Intestine small, duodenum ++++++++++++ M

Intestine small, jejunum ++++++++++++ M

Intestine small, ileum +++++++++++++ 

Liver +++++++++++++++++++++++++ 

Hemangiosarcoma, multiple X X X

Hemangiosarcoma, metastatic, spleen X

Hepatoblastoma X X

Hepatocellular carcinoma X X X X X X

Hepatocellular carcinoma, multiple X X X X XXX X

Hepatocellular adenoma X X X X X X X X X

Hepatocellular adenoma, multiple X X X X X X

Histiocytic sarcoma X

Pancreas +++++++++++++ 

Salivary glands +++++++++++++ 

Stomach, forestomach +++++++++++++ 

Stomach, glandular +++++++++++++ 

Cardiovascular System

Blood vessel +++++++++++++ 

Heart +++++++++++++ 

Alveolar/bronchiolar carcinoma, 

metastatic, lung X

Endocrine System

Adrenal cortex ++++++++++++ M

Adrenal medulla ++++++++++++ M

Islets, pancreatic + + A ++++++++++ 

Parathyroid gland +++ M M + M ++++ M + 

Pituitary gland ++++++ M ++++++ 

Thyroid gland +++++++++++++ 

General Body System

None

Genital System

Epididymis +++++++++++++ 

Preputial gland +++++++++++++ 

Prostate +++++++++++++ 

Seminal vesicle +++++++++++++ 

Testes +++++++++++++ 

Hematopoietic System

Bone marrow +++++++++++++ 

Lymph node + + 

Mediastinal, alveolar/bronchiolar

carcinoma, metastatic, lung X

Mediastinal, histiocytic sarcoma X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 139 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

4444444444445555555555555 

0000000000000000000000000 Total

Carcass ID Number 6667778889995555667777899 Tissues/ 

4582593692462578290467479 Tumors

Alimentary System

Esophagus 13 

Gallbladder 12 

Intestine large, colon 13 

Intestine large, rectum 13 

Intestine large, cecum 13 

Intestine small, duodenum 12 

Intestine small, jejunum 12 

Intestine small, ileum 13 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hemangiosarcoma, multiple 3 

Hemangiosarcoma, metastatic, spleen 1 

Hepatoblastoma X X 4 

Hepatocellular carcinoma X X XXX X X X X X X 17 

Hepatocellular carcinoma, multiple X X X X X X 14 

Hepatocellular adenoma X X X X X X XXX X X X 21 

Hepatocellular adenoma, multiple X X X X X 11 

Histiocytic sarcoma 1 

Pancreas 13 

Salivary glands 13 

Stomach, forestomach 13 

Stomach, glandular 13 

Cardiovascular System

Blood vessel

Heart

Alveolar/bronchiolar carcinoma, 

metastatic, lung

13 

13 

1 

Endocrine System

Adrenal cortex

Adrenal medulla

Islets, pancreatic

Parathyroid gland

Pituitary gland

Thyroid gland

12 

12 

12 

9 

12 

13 

General Body System

None

Genital System

Epididymis

Preputial gland

Prostate

Seminal vesicle

Testes

13 

13 

13 

13 

13 

Hematopoietic System

Bone marrow

Lymph node

Mediastinal, alveolar/bronchiolar

carcinoma, metastatic, lung

Mediastinal, histiocytic sarcoma

13 

2 

1 

1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

140 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

(continued) 

3556666666677777777777777 

Number of Days on Study 2481235578800333333333333 

2080098940726333333334444 

0010000000000000000000000 

Carcass ID Number 6607887957659588889995566 

0708801813691612570353413 

Hematopoietic System (continued) 

Lymph node, mandibular + + M M ++++ MMMM + 

Lymph node, mesenteric + + M + M +++ M + + M + 

Spleen +++++++++++++ 

Hemangiosarcoma X

Thymus ++++++ M M +++++ 

Alveolar/bronchiolar carcinoma, 

metastatic, lung X

Histiocytic sarcoma X

Integumentary System

Mammary gland M M + M + MMMMMMMM

Skin +++++++++++++++++++++++++ 

Subcutaneous tissue, skin, site of

application, histiocytic sarcoma X X

Musculoskeletal System

Bone +++++++++++++ 

Nervous System

Brain +++++++++++++ 

Respiratory System

Lung +++++++++++++ 

Alveolar/bronchiolar adenoma X X X X

Alveolar/bronchiolar carcinoma X

Hepatocellular carcinoma, metastatic, 

liver X X

Histiocytic sarcoma X

Mediastinum, alveolar/bronchiolar

carcinoma, metastatic, lung X

Nose +++++++++++++ 
 #

Trachea +++++++++++++ 
 #

Special Senses System

None

Urinary System

Kidney

Hepatocellular carcinoma, metastatic, 

liver

Urinary bladder

+++++++++++++ 

X

+++++++++++++ 

Systemic Lesions

Multiple organs

Histiocytic sarcoma

+++++++++++++++++++++++++ 

X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 141 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

4444444444445555555555555 

0000000000000000000000000 Total

Carcass ID Number 6667778889995555667777899 Tissues/ 

4582593692462578290467479 Tumors

Hematopoietic System (continued) 

Lymph node, mandibular

Lymph node, mesenteric

Spleen

Hemangiosarcoma

Thymus

Alveolar/bronchiolar carcinoma, 

metastatic, lung

Histiocytic sarcoma

7 

9 

13 

1 

11 

1 

1 

Integumentary System

Mammary gland 2 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Subcutaneous tissue, skin, site of

application, histiocytic sarcoma 2 

Musculoskeletal System

Bone 13 

Nervous System

Brain 13 

Respiratory System

Lung + + + 16 

Alveolar/bronchiolar adenoma 4 

Alveolar/bronchiolar carcinoma 1 

Hepatocellular carcinoma, metastatic, 

liver X X X 5 

Histiocytic sarcoma 1 

Mediastinum, alveolar/bronchiolar

carcinoma, metastatic, lung 1 

Nose 13 

Trachea 13 

Special Senses System

None

Urinary System

Kidney 13 

Hepatocellular carcinoma, metastatic, 

liver 1 

Urinary bladder 13 

Systemic Lesions

Multiple organs + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Histiocytic sarcoma 2 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

142 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

5555666667777777777777777 

Number of Days on Study 0467134881233333333333333 

6812264678233333333333333 

1111111111111111111111111 

Carcass ID Number 4000321011301122223333444 

0369020284955604692678246 

Alimentary System

Esophagus +++++++++++ 

Gallbladder +++++++++++ 

Intestine large, colon +++++++++++ 

Intestine large, rectum +++++++++++ 

Intestine large, cecum +++++++++++ 

Intestine small, duodenum ++++++ M ++++ 

Intestine small, jejunum +++++++++++ 

Intestine small, ileum +++++++++++ 

Liver +++++++++++++++++++++++++ 

Hemangiosarcoma X

Hemangiosarcoma, multiple

Hemangiosarcoma, metastatic, spleen X

Hepatocellular carcinoma X X X X X

Hepatocellular carcinoma, multiple X X X X X X

Hepatocellular adenoma X X X X X

Hepatocellular adenoma, multiple X X X

Osteosarcoma, metastatic, uncertain

primary site

Pancreas +++++++++++ 

Salivary glands +++++++++++ 

Stomach, forestomach +++++++++++ 

Stomach, glandular +++++++++++ 

Cardiovascular System

Blood vessel

Heart

+++++++++++ 

+++++++++++ 

Endocrine System

Adrenal cortex +++++++++++ 

Adrenal medulla +++++++++++ 

Islets, pancreatic +++++++++++ 

Parathyroid gland + + M M + + M + + M + 

Pituitary gland ++++++++ M ++ 

Thyroid gland +++++++++++ 

Follicular cell, adenoma X

General Body System

None

Genital System

Epididymis +++++++++++ 

Preputial gland ++++++ M ++++ 

Prostate +++++++++++ 

Seminal vesicle +++++++++++ 

Testes +++++++++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 143 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

3444444444444445555555555 

1111111111111111111111111 Total

Carcass ID Number 5001222233344440011112344 Tissues/ 

0781357814515791423791338 Tumors

Alimentary System

Esophagus 11 

Gallbladder 11 

Intestine large, colon 11 

Intestine large, rectum 11 

Intestine large, cecum 11 

Intestine small, duodenum 10 

Intestine small, jejunum 11 

Intestine small, ileum 11 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hemangiosarcoma X 2 

Hemangiosarcoma, multiple X X 2 

Hemangiosarcoma, metastatic, spleen 1 

Hepatocellular carcinoma X XXX XXX 12 

Hepatocellular carcinoma, multiple X X X X 10 

Hepatocellular adenoma X X X X X XXX X 14 

Hepatocellular adenoma, multiple X X X X 7 

Osteosarcoma, metastatic, uncertain

primary site X 1 

Pancreas 11 

Salivary glands 11 

Stomach, forestomach 11 

Stomach, glandular 11 

Cardiovascular System

Blood vessel 11 

Heart 11 

Endocrine System

Adrenal cortex

Adrenal medulla

Islets, pancreatic

Parathyroid gland

Pituitary gland

Thyroid gland

Follicular cell, adenoma

11 

11 

11 

7 

10 

11 

1 

General Body System

None

Genital System

Epididymis

Preputial gland

Prostate

Seminal vesicle

Testes

11 

10 

11 

11 

11 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

144 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

Number of Days on Study

5555666667777777777777777 

0467134881233333333333333 

6812264678233333333333333 

Carcass ID Number

1111111111111111111111111 

4000321011301122223333444 

0369020284955604692678246 

Hematopoietic System

Bone marrow

Hemangiosarcoma

Hemangiosarcoma, metastatic, spleen

Lymph node

Lymph node, mandibular

Lymph node, mesenteric

Spleen

Hemangiosarcoma

Hemangiosarcoma, metastatic, 

bone marrow

Thymus

+++++++++++ 

X

X

+ + 

+ M +++++ M + M + 

+++ M +++++++ 

+++++++++++ 

X

X

+ M M +++ M + M + M

Integumentary System

Mammary gland

Skin

Subcutaneous tissue, skin, site of

application, hemangioma

Subcutaneous tissue, skin, site of

application, hemangiosarcoma

M + + MMMMMMMM

+++++++++++++++++++++++++ 

Musculoskeletal System

Bone +++++++++++ 

Nervous System

Brain +++++++++++ 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Hepatocellular carcinoma, metastatic, 

liver

Nose

Trachea

++++++++++++

X X

X

X X X X

+++++++++++ 

+++++++++++ 

+ 

X

Special Senses System

Harderian gland

Adenoma

+ 

X

Urinary System

Kidney

Urinary bladder

+++++++++++ 

+++++++++++ 

Systemic Lesions

Multiple organs

Lymphoma malignant

+++++++++++++++++++++++++ 

X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 145 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

Number of Days on Study

7777777777777777777777777 

3333333333333333333333333 

3444444444444445555555555 

Carcass ID Number

1111111111111111111111111 

5001222233344440011112344 

0781357814515791423791338 

Total

Tissues/ 

Tumors

Hematopoietic System

Bone marrow

Hemangiosarcoma

Hemangiosarcoma, metastatic, spleen

Lymph node

Lymph node, mandibular

Lymph node, mesenteric

Spleen

Hemangiosarcoma

Hemangiosarcoma, metastatic, 

bone marrow

Thymus

+ 

11 

1 

1 

2 

8 

10 

12 

1 

1 

6 

Integumentary System

Mammary gland

Skin

Subcutaneous tissue, skin, site of

application, hemangioma

Subcutaneous tissue, skin, site of

application, hemangiosarcoma

+++++++++++++++++++++++++ 

X

X

2 

50 

1 

1 

Musculoskeletal System

Bone 11 

Nervous System

Brain 11 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Hepatocellular carcinoma, metastatic, 

liver

Nose

Trachea

+

X

+ 

X

15 

2 

1 

7 

11 

11 

Special Senses System

Harderian gland

Adenoma

1 

1 

Urinary System

Kidney

Urinary bladder

11 

11 

Systemic Lesions

Multiple organs

Lymphoma malignant

+++++++++++++++++++++++++ 

X X

50 

4 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

146 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

Number of Days on Study

4555666666777777777777777 

5468233569001123333333333 

7395114841772213333333333 

Carcass ID Number

1112111111111111111111111 

9650985987587965566667777 

3070062551908843436890234 

Alimentary System

Esophagus

Gallbladder

Intestine large, colon

Intestine large, rectum

Intestine large, cecum

Intestine small, duodenum

Intestine small, jejunum

Intestine small, ileum

Liver

Hemangioma

Hemangiosarcoma

Hemangiosarcoma, multiple

Hemangiosarcoma, metastatic, spleen

Hepatoblastoma

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Mesentery

Liposarcoma, multiple

Pancreas

Liposarcoma, metastatic, mesentery

Salivary glands

Stomach, forestomach

Squamous cell papilloma

Stomach, glandular

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

X X

X X

X X X X X X X X

X X X X X X X X

X X X X X X X

X X X X X

+ ++ + 

X

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Cardiovascular System

Blood vessel

Heart

Hemangiosarcoma, metastatic, spleen

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Adrenal medulla +++++++++++++++++++++++++ 

Pheochromocytoma benign X X

Islets, pancreatic +++++++++++++++++++++++++ 

Parathyroid gland M +++++++ M + + M +++++++++ M + + M

Pituitary gland +++++++++++++++++++++++++ 

Thyroid gland +++++++++++++++++++++++++ 

Follicular cell, adenoma

General Body System

None
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 147 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

Number of Days on Study

7777777777777777777777777 

3333333333333333333333333 

3333344444444445555555555 

Carcass ID Number

1111111111111111111111111 

8889955667788995566778899 

2374915175749166825691827 

Total

Tissues/ 

Tumors

Alimentary System

Esophagus

Gallbladder

Intestine large, colon

Intestine large, rectum

Intestine large, cecum

Intestine small, duodenum

Intestine small, jejunum

Intestine small, ileum

Liver

Hemangioma

Hemangiosarcoma

Hemangiosarcoma, multiple

Hemangiosarcoma, metastatic, spleen

Hepatoblastoma

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Mesentery

Liposarcoma, multiple

Pancreas

Liposarcoma, metastatic, mesentery

Salivary glands

Stomach, forestomach

Squamous cell papilloma

Stomach, glandular

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++ M +++++++++++ M +++ 

+++++++++++++++++++++++++ 

X

X

X

X X X X X X X

X X X

X X X X X X

X X X XXXX X X X X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

50 

50 

50 

50 

50 

50 

50 

48 

50 

1 

1 

1 

3 

2 

15 

11 

13 

16 

4 

1 

50 

1 

50 

50 

1 

50 

Cardiovascular System

Blood vessel

Heart

Hemangiosarcoma, metastatic, spleen

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

50 

50 

1 

Endocrine System

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Pheochromocytoma benign 2 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Parathyroid gland ++++ M +++++++++ M ++++++ M + + + 42 

Pituitary gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Follicular cell, adenoma X 1 

General Body System

None
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

148 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

4555666666777777777777777 

Number of Days on Study 5468233569001123333333333 

7395114841772213333333333 

1112111111111111111111111 

Carcass ID Number 9650985987587965566667777 

3070062551908843436890234 

Genital System

Epididymis

Liposarcoma, metastatic, mesentery

Preputial gland

Prostate

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Liposarcoma, metastatic, mesentery

Seminal vesicle

X

+++++++++++++++++++++++++ 

Testes +++++++++++++++++++++++++ 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Lymph node

Lymph node, mandibular

Lymph node, mesenteric

Hemangioma

Hemangiosarcoma, metastatic, spleen

Spleen

Hemangiosarcoma

Liposarcoma, metastatic, mesentery

Thymus

+ 

+++ M M ++++++++ M +++++++++++ 

++++++++ M ++++++++++++++++ 

X

X

+ + M ++++++++++++++++++++++ 

X X

X

+ M + + M + + M + + M + M +++ M + M + + MMM + 

Integumentary System

Mammary gland

Skin

+ M + + MMM + MMMMMMM + M M + MMM + M M

+++++++++++++++++++++++++ 

Subcutaneous tissue, fibrosarcoma X

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Skeletal muscle ++ + 

Hepatocellular carcinoma, metastatic, 

liver X

Liposarcoma, metastatic, mesentery X

Nervous System

Brain +++++++++++++++++++++++++ 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar adenoma, 

multiple

Alveolar/bronchiolar carcinoma

Hemangiosarcoma, metastatic, spleen

Hepatoblastoma, metastatic, liver

Hepatocellular carcinoma, metastatic, 

liver

+++++++++++++++++++++++++ 

X X X X

X

X X

X X XXX X X X X

Nose +++++++++++++++++++++++++ 

Trachea +++++++++++++++++++++++++ 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 149 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

3333344444444445555555555 

1111111111111111111111111 Total

Carcass ID Number 8889955667788995566778899 

2374915175749166825691827 

Tissues/ 

Tumors

Genital System

Epididymis

Liposarcoma, metastatic, mesentery

Preputial gland

Prostate

+++++++++++++++++++++++++ 

+++++++++ M +++++++++++++++ 

+++++++++++++++++++++++++ 

50 

1 

49 

50 

Liposarcoma, metastatic, mesentery

Seminal vesicle +++++++++++++++++++++++++ 

1 

50 

Testes +++++++++++++++++++++++++ 50 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 50 

Lymph node

Lymph node, mandibular

Lymph node, mesenteric

Hemangioma

Hemangiosarcoma, metastatic, spleen

Spleen

Hemangiosarcoma

Liposarcoma, metastatic, mesentery

Thymus

 ++ 

+++ M + + M + + M ++++ M +++++++ M ++ 

++++++++++++ M +++ M ++++++++ 

+++++++++++++++++++++++++ 

X

+ M M + M + + M M ++++++++++ M +++++ 

3 

42 

47 

1 

1 

49 

3 

1 

34 

Integumentary System

Mammary gland

Skin

MMMMMMMM + M + M + MMMM ++++ M + M M

+++++++++++++++++++++++++ 

15 

50 

Subcutaneous tissue, fibrosarcoma 1 

Musculoskeletal System

Bone +++++++++++++++++++++++++ 50 

Skeletal muscle 3 

Hepatocellular carcinoma, metastatic, 

liver 1 

Liposarcoma, metastatic, mesentery 1 

Nervous System

Brain +++++++++++++++++++++++++ 50 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar adenoma, 

multiple

Alveolar/bronchiolar carcinoma

Hemangiosarcoma, metastatic, spleen

Hepatoblastoma, metastatic, liver

Hepatocellular carcinoma, metastatic, 

liver

+++++++++++++++++++++++++ 

X X X

X X

X

X X

50 

7 

2 

1 

1 

2 

11 

Nose +++++++++++++++++++++++++ 50 

Trachea +++++++++++++++++++++++++ 50 
 

 

 

 

 

 

 

 

 

 

 

 

 

150 Sodium Xylenesulfonate, NTP TR 464 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

4555666666777777777777777 

Number of Days on Study 5468233569001123333333333 

7395114841772213333333333 

1112111111111111111111111 

Carcass ID Number 9650985987587965566667777 

3070062551908843436890234 

Special Senses System

Ear + + 

Harderian gland + 

Adenoma X

Carcinoma

Urinary System

Kidney +++++++++++++++++++++++++ 

Hepatocellular carcinoma, metastatic, 

liver X

Renal tubule, adenoma

Renal tubule, carcinoma

Urinary bladder +++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 

Lymphoma malignant X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 151 

TABLE C2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 3333333333333333333333333 

3333344444444445555555555 

1111111111111111111111111 Total

Carcass ID Number 8889955667788995566778899 Tissues/ 

2374915175749166825691827 Tumors

Special Senses System

Ear 2 

Harderian gland + + 3 

Adenoma 1 

Carcinoma X 1 

Urinary System

Kidney +++++++++++++++++++++++++ 50 

Hepatocellular carcinoma, metastatic, 

liver 1 

Renal tubule, adenoma X 1 

Renal tubule, carcinoma X 1 

Urinary bladder +++++++++++++++++++++++++ 50 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 50 

Lymphoma malignant X X X X 6 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

152 Sodium Xylenesulfonate, NTP TR 464 

TABLE C3 

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Liver: Hemangiosarcoma

Overall ratea

Adjusted rateb 5/50 (10%) 

13.6% 

3/50 (6%) 

7.4% 

4/50 (8%) 

10.3% 

2/50 (4%) 

5.7% 

Terminal ratec 3/32 (9%) 1/37 (3%) 4/39 (10%) 2/35 (6%) 

First incidence (days) 

Life table testd

Logistic regression testd

Cochran-Armitage testd

Fisher exact testd

647 

P ̆0.178N

P ̆0.196N

P ̆0.200N

658 

P ̆0.313N

P ̆0.356N

P ̆0.357N

733 (T) 

P ̆0.405N

P ̆0.481N

P ̆0.500N

733 (T) 

P ̆0.196N

P ̆0.213N

P ̆0.218N

Liver: Hepatocellular Adenoma

Overall rate 37/50 (74%) 32/50 (64%) 21/50 (42%) 29/50 (58%) 

Adjusted rate 85.6% 72.5% 51.0% 74.0% 

Terminal rate 26/32 (81%) 25/37 (68%) 19/39 (49%) 25/35 (71%) 

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

564 

P ̆0.029N

P ̆0.043N

P ̆0.046N

322 

P ̆0.080N

P ̆0.194N

P ̆0.194N

612 

P<0.001N

P<0.001N

P ̆0.001N

585 

P ̆0.036N

P ̆0.063N

P ̆0.069N

Liver: Hepatocellular Carcinoma

Overall rate 35/50 (70%) 31/50 (62%) 22/50 (44%) 26/50 (52%) 

Adjusted rate 77.3% 68.8% 47.3% 56.2% 

Terminal rate 22/32 (69%) 23/37 (62%) 15/39 (38%) 15/35 (43%) 

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

494 

P ̆0.037N

P ̆0.030N

P ̆0.030N

610 

P ̆0.144N

P ̆0.264N

P ̆0.263N

506 

P ̆0.005N

P ̆0.009N

P ̆0.007N

457 

P ̆0.056N

P ̆0.051N

P ̆0.050N

Liver: Hepatocellular Adenoma or Carcinoma

Overall rate 46/50 (92%) 41/50 (82%) 31/50 (62%) 45/50 (90%) 

Adjusted rate 93.8% 85.4% 64.3% 93.7% 

Terminal rate 29/32 (91%) 30/37 (81%) 22/39 (56%) 32/35 (91%) 

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

494 

P ̆0.326N

P ̆0.448N

P ̆0.439N

322 

P ̆0.079N

P ̆0.119N

P ̆0.117N

506 

P ̆0.001N

P<0.001N

P<0.001N

457 

P ̆0.290N

P ̆0.501N

P ̆0.500N

Liver: Hepatoblastoma

Overall rate 0/50 (0%) 4/50 (8%) 0/50 (0%) 2/50 (4%) 

Adjusted rate 0.0% 10.8% 0.0% 5.7% 

Terminal rate 0/32 (0%) 4/37 (11%) 0/39 (0%) 2/35 (6%) 

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

—e

P ̆0.466 

P ̆0.466 

P ̆0.444 

733 (T) 

P ̆0.082 

P ̆0.082 

P ̆0.059 

—

—

—

—

733 (T) 

P ̆0.258 

P ̆0.258 

P ̆0.247 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 153 

TABLE C3
#

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Liver: Hepatocellular Carcinoma or Hepatoblastoma

Overall rate 35/50 (70%) 31/50 (62%) 22/50 (44%) 26/50 (52%) 

Adjusted rate 77.3% 68.8% 47.3% 56.2% 

Terminal rate 22/32 (69%) 23/37 (62%) 15/39 (38%) 15/35 (43%) 

First incidence (days) 494 610 506 457 

Life table test P ̆0.037N P ̆0.144N P ̆0.005N P ̆0.056N

Logistic regression test P ̆0.030N P ̆0.264N P ̆0.009N P ̆0.051N

Cochran-Armitage test P ̆0.030N

Fisher exact test P ̆0.263N P ̆0.007N P ̆0.050N

Liver: Hepatocellular Adenoma, Hepatocellular Carcinoma, or Hepatoblastoma

Overall rate 46/50 (92%) 41/50 (82%) 31/50 (62%) 45/50 (90%) 

Adjusted rate 93.8% 85.4% 64.3% 93.7% 

Terminal rate 29/32 (91%) 30/37 (81%) 22/39 (56%) 32/35 (91%) 

First incidence (days) 494 322 506 457 

Life table test P ̆0.326N P ̆0.079N P ̆0.001N P ̆0.290N

Logistic regression test P ̆0.448N P ̆0.119N P<0.001N P ̆0.501N

Cochran-Armitage test P ̆0.439N

Fisher exact test P ̆0.117N P<0.001N P ̆0.500N

Lung: Alveolar/bronchiolar Adenoma

Overall rate 13/50 (26%) 4/16 (25%) f 2/15 (13%) f 9/50 (18%) 

Adjusted rate 40.6% 22.9% 

Terminal rate 13/32 (41%) 6/35 (17%) 

First incidence (days) 733 (T) 543 

Life table test P ̆0.160N

Logistic regression test P ̆0.212N

Fisher exact test P ̆0.235N

Lung: Alveolar/bronchiolar Carcinoma

Overall rate 2/50 (4%) 1/16 (6%) f 1/15 (7%) f 1/50 (2%) 

Adjusted rate 4.4% 2.2% 

Terminal rate 0/32 (0%) 0/35 (0%) 

First incidence (days) 575 631 

Life table test P ̆0.508N

Logistic regression test P ̆0.470N

Fisher exact test P ̆0.500N

Lung: Alveolar/bronchiolar Adenoma or Carcinoma

Overall rate 15/50 (30%) 5/16 (31%) f 3/15 (20%) f 10/50 (20%) 

Adjusted rate 43.2% 24.6% 

Terminal rate 13/32 (41%) 6/35 (17%) 

First incidence (days) 575 543 

Life table test P ̆0.128N

Logistic regression test P ̆0.175N

Fisher exact test P ̆0.178N
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

154 Sodium Xylenesulfonate, NTP TR 464 

TABLE C3
#

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Spleen: Hemangiosarcoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

2/50 (4%) 

5.5% 

1/32 (3%) 

686 

1/13 (8%) f 1/12 (8%) f 3/49 (6%) 

8.0% 

2/35 (6%) 

691 

P ̆0.531 

P ̆0.498 

P ̆0.490 

Thyroid Gland (Follicular Cell): Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

2/50 (4%) 

5.9% 

1/32 (3%) 

702 

0/13 (0%) f 1/11 (9%) f 1/50 (2%) 

2.9% 

1/35 (3%) 

733 (T) 

P ̆0.464N

P ̆0.490N

P ̆0.500N

All Organs: Hemangioma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

3/50 (6%) 

8.0% 

1/32 (3%) 

575 

0/50 (0%) f 1/50 (2%) f 2/50 (4%) 

4.4% 

0/35 (0%) 

631 

P ̆0.483N

P ̆0.484N

P ̆0.500N

All Organs: Hemangiosarcoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

6/50 (12%) 

15.7% 

3/32 (9%) 

647 

4/50 (8%) f 6/50 (12%) f 5/50 (10%) 

13.6% 

4/35 (11%) 

691 

P ̆0.458N

P ̆0.496N

P ̆0.500N

All Organs: Hemangioma or Hemangiosarcoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

9/50 (18%) 

22.7% 

4/32 (13%) 

575 

4/50 (8%) f 7/50 (14%) f 7/50 (14%) 

17.4% 

4/35 (11%) 

631 

P ̆0.356N

P ̆0.393N

P ̆0.393N
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

155 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE C3
#

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

All Organs: Malignant Lymphoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

5/50 (10%) 

12.3% 

0/32 (0%) 

651 

0/50 (0%) f 4/50 (27%) f 6/50 (12%) 

15.6% 

4/35 (11%) 

631 

P ̆0.550 

P ̆0.500 

P ̆0.500 

All Organs: Benign Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

43/50 (86%) 

95.4% 

30/32 (94%) 

564 

33/50 (66%)f 24/50 (48%)f 37/50 (74%) 

83.9% 

28/35 (80%) 

543 

P ̆0.071N

P ̆0.100N

P ̆0.105N

All Organs: Malignant Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

39/50 (78%) 

81.1% 

23/32 (72%) 

494 

33/50 (66%)f 29/50 (58%)f 33/50 (66%) 

68.7% 

20/35 (57%) 

457 

P ̆0.129N

P ̆0.136N

P ̆0.133N

All Organs: Benign or Malignant Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

49/50 (98%) 

98.0% 

31/32 (97%) 

494 

41/50 (82%)f 37/50 (74%)f 47/50 (94%) 

95.9% 

33/35 (94%) 

457 

P ̆0.218N

P ̆0.305N

P ̆0.309N

(T)Terminal sacrifice a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for liver, lung, spleen, 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 dosed group incidence are the P values corresponding to pairwise

comparisons between the controls and that dosed 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 a dose group is indicated by N. e Not applicable; no neoplasms in animal group f	 Tissues (except skin) were examined microscopically only in those animals dying prior to terminal sacrifice or when it was observed to be abnormal at

necropsy; thus statistical comparisons with the controls are not applicable. 
 

 

 

 

 

 

 

 

 

156 Sodium Xylenesulfonate, NTP TR 464 

TABLE C4 

Historical a Incidence of Hepatocellular Neoplasms in Control Male B6C3F1 Mice

Incidence in Controls

Study Adenoma Carcinoma Adenoma

or Carcinoma

Historical Incidence at Battelle Columbus: Dermal (Acetone) Studies

4-Vinyl-1-cyclohexene Diepoxide 18/50 6/50 23/50 

Triethanolamine 27/50 15/50 31/50 

Overall Historical Incidence: Dermal (Acetone) Studies

Total 51/150 (34.0%) 25/150 (16.7%) 63/150 (42.0%) 

Standard deviation 21.1% 11.7% 22.3% 

Range 12%-54% 8%-30% 18%-62% 

Historical Incidence at Battelle Columbus: Dermal (Ethanol) Study

Benzethonium Chloride 24/50 (48.0%) 10/50 (20.0%) 29/50 (58.0%) 

Historical Incidence at Battelle Columbus: Feed Studies

4,4-Thiobis(6-t-butyl-m-cresol) 17/50 11/50 25/50 

5,5-Diphenylhydantoin 19/50 13/50 29/50 

Pentachlorophenol (Dowicide EC-7) 5/35 1/35 6/35 

Ethylene Thiourea 11/49 13/49 20/49 

Polybrominated Biphenyls (Firemaster FF-1®) 9/50 8/50 16/50 

Manganese (II) Sulfate Monohydrate 30/50 9/50 34/50 

Oxazepam 17/49 9/49 23/49 

Technical Grade Pentachlorophenol 5/32 2/32 7/32 

Triamterene 17/50 5/50 20/50 

Triamterene 21/50 9/50 25/50 

Tricresyl Phosphate 18/52 15/52 28/52 

Overall Historical Incidence: Feed Studies

Total 413/1,465 (29.2%) 252/1,465 (17.2%) 596/1,465 (40.7%) 

Standard deviation 14.2% 7.1% 14.5% 

Range 4%-60% 3%-29% 10%-68% 

a Data as of 12 May 1995 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 157 

TABLE C5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Moribund 7 3 8 6 

Natural deaths 11 10 3 9 

Survivors

Terminal sacrifice 32 37 39 35 

Animals examined microscopically 50 50 50 50 

Alimentary System

Intestine small, duodenum (50) (12) (10) (50) 

Necrosis 1 (2%) 

Ulcer 1 (2%) 

Liver (50) (50) (50) (50) 

Angiectasis 2 (4%) 2 (4%) 1 (2%) 

Basophilic focus 2 (4%) 6 (12%) 3 (6%) 4 (8%) 

Clear cell focus 4 (8%) 3 (6%) 3 (6%) 8 (16%) 

Eosinophilic focus 13 (26%) 11 (22%) 12 (24%) 9 (18%) 

Hematopoietic cell proliferation 3 (6%) 1 (2%) 

Hemorrhage 1 (2%) 

Infarct 1 (2%) 

Inflammation, chronic active 46 (92%) 44 (88%) 41 (82%) 43 (86%) 

Mixed cell focus 3 (6%) 4 (8%) 5 (10%) 7 (14%) 

Necrosis 8 (16%) 5 (10%) 2 (4%) 8 (16%) 

Thrombosis 1 (2%) 1 (2%) 

Vacuolization cytoplasmic 2 (4%) 1 (2%) 1 (2%) 

Bile duct, hyperplasia 43 (86%) 41 (82%) 36 (72%) 37 (74%) 

Kupffer cell, pigmentation 1 (2%) 

Mesentery (5) (4) 

Hemorrhage 1 (25%) 

Artery, inflammation, chronic active 1 (20%) 1 (25%) 

Fat, inflammation 1 (20%) 

Vein, thrombosis 1 (20%) 1 (25%) 

Pancreas (50) (13) (11) (50) 

Atrophy 3 (6%) 1 (9%) 4 (8%) 

Cytoplasmic alteration 4 (8%) 

Necrosis 1 (2%) 1 (9%) 

Artery, inflammation, chronic active 2 (4%) 

Salivary glands (50) (13) (11) (50) 

Atrophy 1 (2%) 

Necrosis 1 (9%) 

Stomach, forestomach (50) (13) (11) (50) 

Cyst 1 (2%) 

Hyperplasia, focal 1 (2%) 1 (9%) 2 (4%) 

Ulcer 2 (4%) 

Stomach, glandular (50) (13) (11) (50) 

Erosion 1 (2%) 

Mineralization 1 (2%) 

Artery, inflammation, chronic active 1 (2%) 

a Number of animals examined microscopically at the site and the number of animals with lesion
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

158 Sodium Xylenesulfonate, NTP TR 464 

TABLE C5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Cardiovascular System

Blood vessel (50) (13) (11) (50) 

Aorta, inflammation, chronic active 1 (2%) 1 (9%) 1 (2%) 

Heart (50) (13) (11) (50) 

Degeneration 2 (4%) 1 (8%) 1 (9%) 1 (2%) 

Mineralization 1 (2%) 2 (4%) 

Artery, inflammation, chronic active 1 (2%) 1 (2%) 

Atrium, thrombosis 1 (2%) 1 (8%) 1 (9%) 1 (2%) 

Ventricle, thrombosis 1 (8%) 

Endocrine System

Adrenal cortex (50) (12) (11) (50) 

Accessory adrenal cortical nodule 2 (4%) 1 (9%) 1 (2%) 

Degeneration, cystic 1 (2%) 

Hemorrhage 1 (2%) 

Hyperplasia 11 (22%) 1 (8%) 1 (9%) 19 (38%) 

Inflammation, chronic active 1 (2%) 

Capsule, hyperplasia, adenomatous 4 (8%) 

Adrenal medulla (50) (12) (11) (50) 

Hyperplasia 2 (4%) 2 (17%) 1 (9%) 1 (2%) 

Islets, pancreatic (50) (12) (11) (50) 

Hyperplasia 9 (18%) 2 (18%) 9 (18%) 

Parathyroid gland (46) (9) (7) (42) 

Cyst 1 (2%) 

Pituitary gland (49) (12) (10) (50) 

Cyst 2 (4%) 2 (4%) 

Pars distalis, hyperplasia 2 (4%) 2 (4%) 

Pars intermedia, hyperplasia 2 (4%) 

Thyroid gland (50) (13) (11) (50) 

Follicular cell, hyperplasia 6 (12%) 2 (15%) 10 (20%) 

General Body System

None

Genital System

Epididymis (50) (13) (11) (50) 

Granuloma sperm 2 (4%) 2 (4%) 

Artery, inflammation, chronic active 1 (9%) 1 (2%) 

Preputial gland (50) (13) (10) (49) 

Inflammation, chronic active 1 (2%) 

Duct, cyst 7 (14%) 1 (8%) 4 (40%) 6 (12%) 

Prostate (50) (13) (11) (50) 

Degeneration 1 (9%) 

Artery, inflammation, chronic active 1 (9%) 1 (2%) 

Seminal vesicle (50) (13) (11) (50) 

Degeneration 1 (9%) 

Testes (50) (13) (11) (50) 

Atrophy 2 (4%) 

Artery, inflammation, chronic active 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 159 

TABLE C5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Hematopoietic System

Bone marrow (50) (13) (11) (50) 

Angiectasis 1 (2%) 

Hyperplasia 17 (34%) 3 (23%) 4 (36%) 23 (46%) 

Infiltration cellular, mast cell 1 (2%) 

Infiltration cellular, plasma cell 1 (2%) 

Myelofibrosis 1 (2%) 

Thrombosis 1 (2%) 

Lymph node (3) (2) (2) (3) 

Mediastinal, angiectasis 1 (33%) 

Mediastinal, hematopoietic cell proliferation 1 (33%) 

Lymph node, mesenteric (50) (9) (10) (47) 

Angiectasis 1 (2%) 

Hematopoietic cell proliferation 1 (2%) 1 (10%) 

Hyperplasia, histiocytic 1 (2%) 

Hyperplasia, lymphoid 1 (2%) 1 (10%) 

Hyperplasia, plasma cell 1 (2%) 

Spleen (50) (13) (12) (49) 

Hematopoietic cell proliferation 27 (54%) 8 (62%) 8 (67%) 25 (51%) 

Lymphoid follicle, depletion cellular 1 (2%) 

Lymphoid follicle, hyperplasia 1 (2%) 

Thymus (37) (11) (6) (34) 

Atrophy 9 (24%) 5 (45%) 2 (33%) 6 (18%) 

Integumentary System

Skin (50) (50) (50) (50) 

Epidermis, skin, site of application, exudate 1 (2%) 1 (2%) 1 (2%) 

Epidermis, skin, site of application, 

hyperkeratosis 1 (2%) 

Epidermis, skin, site of application, 

hyperplasia 1 (2%) 4 (8%) 5 (10%) 

Epidermis, skin, site of application, ulcer 1 (2%) 

Subcutaneous tissue, edema 1 (2%) 1 (2%) 1 (2%) 

Subcutaneous tissue, inflammation, 

suppurative 1 (2%) 

Musculoskeletal System

None

Nervous System

Brain (50) (13) (11) (50) 

Inflammation 1 (9%) 

Artery, inflammation, chronic active 1 (9%) 

Corpus callosum, degeneration 1 (2%) 

Neuron, necrosis 1 (8%) 1 (9%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

160 Sodium Xylenesulfonate, NTP TR 464 

TABLE C5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Respiratory System

Lung (50) (16) (15) (50) 

Inflammation 1 (7%) 

Pigmentation, hemosiderin 1 (7%) 

Thrombosis 1 (7%) 

Alveolar epithelium, hyperplasia 2 (13%) 1 (7%) 2 (4%) 

Mediastinum, angiectasis 1 (2%) 

Nose (50) (13) (11) (50) 

Inflammation, suppurative 1 (2%) 

Special Senses System

Ear (2) 

Inflammation 1 (50%) 

Harderian gland (2) (1) (3) 

Hyperplasia 1 (50%) 1 (33%) 

Urinary System

Kidney (50) (13) (11) (50) 

Cyst 5 (10%) 8 (16%) 

Glomerulosclerosis 2 (4%) 1 (8%) 4 (8%) 

Hydronephrosis 1 (2%) 

Infarct 1 (2%) 1 (2%) 

Mineralization 4 (8%) 

Nephropathy 43 (86%) 10 (77%) 8 (73%) 43 (86%) 

Pigmentation, hemosiderin 1 (2%) 1 (9%) 1 (2%) 

Artery, inflammation, chronic active 1 (9%) 1 (2%) 

Glomerulus, infarct 1 (2%) 

Glomerulus, thrombosis 1 (9%) 

Renal tubule, hyperplasia 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

161 

APPENDIX D
 

SUMMARY OF LESIONS IN FEMALE MICE
 

IN THE 2-YEAR DERMAL STUDY
 

OF SODIUM XYLENESULFONATE
 

TABLE D1	# Summary of the Incidence of Neoplasms in Female Mice

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 163 
 #

TABLE D2 Individual Animal Tumor Pathology of Female Mice
 

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 168 
 #

TABLE D3 Statistical Analysis of Primary Neoplasms in Female Mice
 

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 190 
 #

TABLE D4 Historical Incidence of Hepatocellular Neoplasms in Control Female B6C3F1 Mice . . . . 195 
 #

TABLE D5	# Summary of the Incidence of Nonneoplastic Lesions in Female Mice
 

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 196 
 #
162 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 163 

TABLE D1 

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Moribund 12 7 9 6 

Natural deaths 7 10 9 8 

Survivors

Terminal sacrifice 31 32 32 36 

Missing 1 

Animals examined microscopically 50 49 50 50 

Alimentary System

Gallbladder (49) (17) (18) (50) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (6%) 

Intestine large, cecum (50) (16) (18) (50) 

Leiomyosarcoma 1 (6%) 

Intestine small, jejunum (49) (16) (18) (50) 

Fibrosarcoma, metastatic, skin 1 (2%) 

Leiomyosarcoma 1 (6%) 

Liver (50) (49) (50) (50) 

Hemangioma 2 (4%) 

Hemangiosarcoma 1 (2%) 1 (2%) 

Hepatocellular carcinoma 9 (18%) 10 (20%) 7 (14%) 8 (16%) 

Hepatocellular carcinoma, multiple 1 (2%) 3 (6%) 2 (4%) 

Hepatocellular adenoma 10 (20%) 8 (16%) 14 (28%) 19 (38%) 

Hepatocellular adenoma, multiple 8 (16%) 9 (18%) 4 (8%) 9 (18%) 

Hepatocholangiocarcinoma 1 (2%) 1 (2%) 2 (4%) 

Histiocytic sarcoma 3 (6%) 2 (4%) 2 (4%) 2 (4%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (2%) 

Mesentery (7) (6) (5) (12) 

Fibrosarcoma, metastatic, skin 1 (8%) 

Hemangioma 1 (8%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (14%) 1 (20%) 2 (17%) 

Histiocytic sarcoma 1 (14%) 1 (17%) 1 (20%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (20%) 

Pancreas (50) (17) (18) (50) 

Fibrosarcoma, metastatic, skin 1 (2%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (2%) 1 (6%) 

Histiocytic sarcoma 1 (2%) 1 (6%) 1 (6%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (6%) 

Salivary glands (50) (17) (18) (50) 

Stomach, forestomach (50) (17) (18) (50) 

Squamous cell papilloma 2 (12%) 

Stomach, glandular (50) (17) (18) (50) 

Carcinoma 1 (6%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (6%) 

Histiocytic sarcoma 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

164 Sodium Xylenesulfonate, NTP TR 464 

TABLE D1
#

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Cardiovascular System

Blood vessel (50) (17) (18) (50) 

Aorta, hepatocholangiocarcinoma, metastatic, 

liver 1 (6%) 

Heart (50) (17) (18) (50) 

Alveolar/bronchiolar carcinoma, metastatic, 

lung 1 (6%) 

Histiocytic sarcoma 2 (4%) 1 (6%) 

Endocrine System

Adrenal cortex (50) (17) (18) (50) 

Hepatocholangiocarcinoma, metastatic, liver 1 (6%) 

Capsule, adenoma 1 (2%) 1 (2%) 

Adrenal medulla (50) (17) (18) (48) 

Pheochromocytoma benign 1 (2%) 

Islets, pancreatic (50) (17) (18) (50) 

Adenoma 1 (2%) 1 (6%) 

Carcinoma 1 (2%) 

Pituitary gland (50) (17) (18) (49) 

Meningioma malignant, metastatic, brain 1 (6%) 

Pars distalis, adenoma 7 (14%) 3 (18%) 1 (6%) 5 (10%) 

Pars intermedia, adenoma 1 (2%) 2 (4%) 

Pars intermedia, carcinoma 1 (2%) 

Thyroid gland (50) (48) (50) (50) 

Follicular cell, adenoma 1 (2%) 5 (10%) 1 (2%) 6 (12%) 

Follicular cell, adenoma, multiple 1 (2%) 

Follicular cell, carcinoma 1 (2%) 

General Body System

None

Genital System

Clitoral gland (49) (15) (15) (46) 

Hepatocholangiocarcinoma, metastatic, liver 1 (2%) 

Ovary (50) (17) (18) (49) 

Cystadenoma 1 (6%) 1 (2%) 

Fibrosarcoma, metastatic, skin 1 (2%) 

Hemangioma 1 (6%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (6%) 

Histiocytic sarcoma 1 (2%) 1 (6%) 1 (6%) 1 (2%) 

Teratoma benign 1 (2%) 

Uterus (50) (17) (18) (50) 

Hemangiosarcoma 1 (2%) 

Histiocytic sarcoma 2 (12%) 1 (2%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (6%) 

Polyp stromal 3 (6%) 1 (6%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 165 

TABLE D1
#

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Hematopoietic System

Bone marrow (50) (17) (18) (50) 

Histiocytic sarcoma 1 (2%) 2 (12%) 

Lymph node (7) (3) (6) (6) 

Bronchial, alveolar/bronchiolar carcinoma, 

metastatic, lung 1 (17%) 

Inguinal, histiocytic sarcoma 1 (17%) 

Lumbar, histiocytic sarcoma 2 (67%) 2 (33%) 

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver 1 (14%) 1 (17%) 1 (17%) 

Mediastinal, histiocytic sarcoma 1 (14%) 1 (33%) 1 (17%) 

Pancreatic, histiocytic sarcoma 1 (17%) 

Renal, hepatocholangiocarcinoma, metastatic, 

liver 1 (17%) 

Renal, histiocytic sarcoma 1 (17%) 1 (17%) 

Thoracic, fibrosarcoma, metastatic, skin 1 (14%) 

Lymph node, mandibular (49) (15) (15) (48) 

Carcinoma, metastatic, harderian gland 1 (7%) 

Histiocytic sarcoma 2 (4%) 1 (2%) 

Lymph node, mesenteric (48) (17) (17) (50) 

Fibrosarcoma, metastatic, skin 1 (2%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (6%) 2 (4%) 

Histiocytic sarcoma 2 (4%) 2 (12%) 1 (6%) 2 (4%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (6%) 

Spleen (50) (17) (18) (50) 

Hemangiosarcoma 1 (6%) 2 (4%) 

Histiocytic sarcoma 3 (6%) 1 (6%) 1 (2%) 

Thymus (38) (16) (16) (44) 

Alveolar/bronchiolar carcinoma, metastatic, 

lung 1 (6%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (6%) 2 (5%) 

Histiocytic sarcoma 1 (3%) 1 (2%) 

Integumentary System

Mammary gland (50) (17) (16) (50) 

Carcinoma 1 (2%) 

Skin (50) (49) (50) (50) 

Subcutaneous tissue, fibrosarcoma 1 (2%) 1 (2%) 

Subcutaneous tissue, hemangiosarcoma 1 (2%) 

Subcutaneous tissue, skin, site of

application, fibrosarcoma 1 (2%) 1 (2%) 

Musculoskeletal System

Bone (50) (17) (18) (50) 

Osteosarcoma 1 (2%) 

Skeletal muscle (1) (3) 

Fibrosarcoma, metastatic, skin 1 (33%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (100%) 2 (67%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

166 Sodium Xylenesulfonate, NTP TR 464 

TABLE D1
#

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Nervous System

Brain (50) (17) (18) (50) 

Carcinoma, metastatic, harderian gland 1 (6%) 

Histiocytic sarcoma 1 (2%) 

Meningioma malignant 1 (6%) 

Respiratory System

Lung (50) (18) (18) (50) 

Alveolar/bronchiolar adenoma 4 (8%) 1 (6%) 2 (11%) 6 (12%) 

Alveolar/bronchiolar carcinoma 2 (4%) 1 (6%) 

Carcinoma, metastatic, harderian gland 1 (2%) 2 (11%) 1 (2%) 

Fibrosarcoma, metastatic, skin 1 (2%) 

Hepatocellular carcinoma, metastatic, liver 2 (4%) 2 (11%) 1 (6%) 3 (6%) 

Hepatocholangiocarcinoma, metastatic, liver 1 (2%) 1 (6%) 2 (4%) 

Histiocytic sarcoma 3 (6%) 1 (6%) 2 (4%) 

Leiomyosarcoma, metastatic, intestine small, 

jejunum 1 (6%) 

Leiomyosarcoma, metastatic, intestine large, 

cecum 1 (6%) 

Squamous cell carcinoma, metastatic, urinary

bladder 1 (2%) 

Mediastinum, alveolar/bronchiolar carcinoma, 

metastatic, lung 1 (6%) 

Nose (50) (17) (18) (50) 

Histiocytic sarcoma 1 (2%) 

Special Senses System

Eye (1) (2) 

Carcinoma, metastatic, harderian gland 1 (50%) 

Harderian gland (5) (2) (1) (3) 

Adenoma 2 (40%) 1 (33%) 

Carcinoma 1 (20%) 2 (100%) 1 (100%) 2 (67%) 

Bilateral, adenoma 1 (20%) 

Urinary System

Kidney (50) (17) (18) (50) 

Hepatocholangiocarcinoma, metastatic, liver 1 (2%) 

Histiocytic sarcoma 3 (6%) 2 (12%) 

Urinary bladder (50) (17) (18) (50) 

Squamous cell carcinoma 1 (2%) 

Systemic Lesions

Multiple organsb (50) (49) (50) (50) 

Histiocytic sarcoma 3 (6%) 2 (4%) 2 (4%) 2 (4%) 

Lymphoma malignant 8 (16%) 3 (6%) 3 (6%) 5 (10%) 
 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 167 

TABLE D1
#

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate (continued)
#

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Neoplasm Summary

Total animals with primary neoplasmsc 45 34 31 44 

Total primary neoplasms 71 54 44 82 

Total animals with benign neoplasms 29 22 20 35 

Total benign neoplasms 43 30 24 52 

Total animals with malignant neoplasms 25 22 18 26 

Total malignant neoplasms 28 24 20 30 

Total animals with metastatic neoplasms 5 5 5 8 

Total metastatic neoplasms 9 8 24 25 

a

b

Number of animals examined microscopically at the site and the number of animals with neoplasm

Number of animals with any tissue examined microscopically c Primary neoplasms: all neoplasms except metastatic neoplasms
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

168 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 

Vehicle Control

2444456666666667777777777 

Number of Days on Study 9569982225577890223444444 

2315990119909021264000000 

2222222222222222222222222 

Carcass ID Number 0340442231311144403001222 

3285066101756017995684235 

Alimentary System

Esophagus +++++++++++++++++++++++++ 

Gallbladder + + M ++++++++++++++++++++++ 

Intestine large, colon +++++++++++++++++++++++++ 

Intestine large, rectum +++++++++++++++++++++++++ 

Intestine large, cecum +++++++++++++++++++++++++ 

Intestine small, duodenum +++++++++++++++++++++++++ 

Intestine small, jejunum ++++++++++++++++++ M ++++++ 

Intestine small, ileum +++++++++++++++++++++++++ 

Liver +++++++++++++++++++++++++ 

Hemangioma X

Hepatocellular carcinoma X X X X X X

Hepatocellular carcinoma, multiple X

Hepatocellular adenoma X X X

Hepatocellular adenoma, multiple X

Hepatocholangiocarcinoma X

Histiocytic sarcoma X X X

Mesentery + + + + + 

Hepatocholangiocarcinoma, metastatic, 

liver X

Histiocytic sarcoma X

Pancreas +++++++++++++++++++++++++ 

Hepatocholangiocarcinoma, metastatic, 

liver X

Histiocytic sarcoma X

Salivary glands +++++++++++++++++++++++++ 

Stomach, forestomach +++++++++++++++++++++++++ 

Stomach, glandular +++++++++++++++++++++++++ 

Histiocytic sarcoma X

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Histiocytic sarcoma X X

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Capsule, adenoma

Adrenal medulla +++++++++++++++++++++++++ 

Islets, pancreatic +++++++++++++++++++++++++ 

Adenoma X

Carcinoma X

Parathyroid gland +++++++ M + M M + M M ++++ M ++++ M + 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X X

Pars intermedia, adenoma

Thyroid gland +++++++++++++++++++++++++ 

Follicular cell, adenoma X

Follicular cell, adenoma, multiple X

+: Tissue examined microscopically M: Missing tissue X: Lesion present

A: Autolysis precludes examination I: Insufficient tissue Blank: Not examined
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 169 

TABLE D2
#

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0000111111112222222222222 

2222222222222222222222222 Total

Carcass ID Number 3344002233450011111222334 Tissues/ 

1835144846202723789079394 Tumors

Alimentary System

Esophagus +++++++++++++++++++++++++ 50 

Gallbladder +++++++++++++++++++++++++ 49 

Intestine large, colon +++++++++++++++++++++++++ 50 

Intestine large, rectum +++++++++++++++++++++++++ 50 

Intestine large, cecum +++++++++++++++++++++++++ 50 

Intestine small, duodenum +++++++++++++++++++++++++ 50 

Intestine small, jejunum +++++++++++++++++++++++++ 49 

Intestine small, ileum +++++++++++++++++++++++++ 50 

Liver +++++++++++++++++++++++++ 50 

Hemangioma X 2 

Hepatocellular carcinoma X X X 9 

Hepatocellular carcinoma, multiple 1 

Hepatocellular adenoma X X X X X X X 10 

Hepatocellular adenoma, multiple X X X X X X X 8 

Hepatocholangiocarcinoma 1 

Histiocytic sarcoma 3 

Mesentery + + 7 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Histiocytic sarcoma 1 

Pancreas +++++++++++++++++++++++++ 50 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Histiocytic sarcoma 1 

Salivary glands +++++++++++++++++++++++++ 50 

Stomach, forestomach +++++++++++++++++++++++++ 50 

Stomach, glandular +++++++++++++++++++++++++ 50 

Histiocytic sarcoma 1 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 50 

Heart +++++++++++++++++++++++++ 50 

Histiocytic sarcoma 2 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 50 

Capsule, adenoma X 1 

Adrenal medulla +++++++++++++++++++++++++ 50 

Islets, pancreatic +++++++++++++++++++++++++ 50 

Adenoma 1 

Carcinoma 1 

Parathyroid gland M ++++++ M ++++++ M +++ M + M ++++ 38 

Pituitary gland +++++++++++++++++++++++++ 50 

Pars distalis, adenoma X X X X X 7 

Pars intermedia, adenoma X 1 

Thyroid gland +++++++++++++++++++++++++ 50 

Follicular cell, adenoma 1 

Follicular cell, adenoma, multiple 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

170 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2
#

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

2444456666666667777777777 

Number of Days on Study 9569982225577890223444444 

2315990119909021264000000 

2222222222222222222222222 

Carcass ID Number 0340442231311144403001222 

3285066101756017995684235 

General Body System

None

Genital System

Clitoral gland

Ovary

Histiocytic sarcoma

Teratoma benign

Uterus

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

Polyp stromal X

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Histiocytic sarcoma

Lymph node

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver

Mediastinal, histiocytic sarcoma

Thoracic, fibrosarcoma, metastatic, 

skin

+

X

X

+

X

+ ++

X

+ 

Lymph node, mandibular

Histiocytic sarcoma

Lymph node, mesenteric

Histiocytic sarcoma

Spleen

Histiocytic sarcoma

Thymus

Histiocytic sarcoma

+++++++++++++++ M +++++++++ 

X X

+++++++++++++++++++++++++ 

X X

+++++++++++++++++++++++++ 

X X X

+++ M + + M + + M M + + M + + M + M + + M M ++ 

X

Integumentary System

Mammary gland

Skin

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Subcutaneous tissue, fibrosarcoma X

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Osteosarcoma

Nervous System

Brain +++++++++++++++++++++++++ 

Histiocytic sarcoma

Peripheral nerve

Spinal cord

 + 

+ 

X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 171 

TABLE D2
#

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0000111111112222222222222 

2222222222222222222222222 Total

Carcass ID Number 3344002233450011111222334 

1835144846202723789079394 

Tissues/ 

Tumors

General Body System

None

Genital System

Clitoral gland

Ovary

Histiocytic sarcoma

Teratoma benign

Uterus

++++++++ M ++++++++++++++++ 

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

49 

50 

1 

1 

50 

Polyp stromal X X 3 

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 50 

Histiocytic sarcoma

Lymph node

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver

Mediastinal, histiocytic sarcoma

Thoracic, fibrosarcoma, metastatic, 

skin

+ 

1 

7 

1 

1 

1 

Lymph node, mandibular

Histiocytic sarcoma

Lymph node, mesenteric

Histiocytic sarcoma

Spleen

Histiocytic sarcoma

Thymus

Histiocytic sarcoma

+++++++++++++++++++++++++ 

+++++++++++++++++ M +++ M +++ 

+++++++++++++++++++++++++ 

+ M ++++++ M ++++++ M +++++++++ 

49 

2 

48 

2 

50 

3 

38 

1 

Integumentary System

Mammary gland

Skin

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

50 

Subcutaneous tissue, fibrosarcoma 1 

Musculoskeletal System

Bone +++++++++++++++++++++++++ 50 

Osteosarcoma X 1 

Nervous System

Brain +++++++++++++++++++++++++ 50 

Histiocytic sarcoma

Peripheral nerve

Spinal cord

1 

1 

1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

172 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2
#

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

2444456666666667777777777 

Number of Days on Study 9569982225577890223444444 

2315990119909021264000000 

2222222222222222222222222 

Carcass ID Number 0340442231311144403001222 

3285066101756017995684235 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Carcinoma, metastatic, harderian

gland

Fibrosarcoma, metastatic, skin

Hepatocellular carcinoma, metastatic, 

liver

+++++++++++++++++++++++++ 

X X X

X

X

X X

Hepatocholangiocarcinoma, metastatic, 

liver X

Histiocytic sarcoma

Nose

X X X

+++++++++++++++++++++++++ 

Trachea +++++++++++++++++++++++++ 

Special Senses System

Eye

Harderian gland

Adenoma

+

 + 

+

X

 ++ 

Carcinoma X

Bilateral, adenoma X

Urinary System

Kidney

Histiocytic sarcoma

Urinary bladder

+++++++++++++++++++++++++ 

X X X

+++++++++++++++++++++++++ 

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Lymphoma malignant

+++++++++++++++++++++++++ 

X X X

X X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 173 

TABLE D2
#

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate:
#

Vehicle Control (continued)
#

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0000111111112222222222222 

2222222222222222222222222 Total

Carcass ID Number 3344002233450011111222334 

1835144846202723789079394 

Tissues/ 

Tumors

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Carcinoma, metastatic, harderian

gland

Fibrosarcoma, metastatic, skin

Hepatocellular carcinoma, metastatic, 

liver

+++++++++++++++++++++++++ 

X

X X

50 

4 

2 

1 

1 

2 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Histiocytic sarcoma

Nose +++++++++++++++++++++++++ 

3 

50 

Trachea +++++++++++++++++++++++++ 50 

Special Senses System

Eye

Harderian gland

Adenoma

+ 

X

1 

5 

2 

Carcinoma 1 

Bilateral, adenoma 1 

Urinary System

Kidney

Histiocytic sarcoma

Urinary bladder

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

3 

50 

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Lymphoma malignant

+++++++++++++++++++++++++ 

X X X X X

50 

3 

8 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

174 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

034445666666677777777777 

Number of Days on Study 231463023456711334444444 

471678456296428150000000 

222222222222222222222222 

Carcass ID Number 788597785987598965556666 

169550777038242651391234 

Alimentary System

Esophagus ++++++++ M ++++++++ 

Gallbladder +++++++++++++++++ 

Intestine large, colon +++++++++++++++++ 

Intestine large, rectum +++++++++++++++++ 

Intestine large, cecum +++++++++++ M +++++ 

Intestine small, duodenum +++++++++++++++++ 

Intestine small, jejunum +++++++++++ M +++++ 

Intestine small, ileum +++++++++++++++++ 

Liver ++++++++++++++++++++++++ 

Hemangiosarcoma

Hepatocellular carcinoma X X XXX X X

Hepatocellular carcinoma, multiple

Hepatocellular adenoma X

Hepatocellular adenoma, multiple X X X X X

Histiocytic sarcoma X X

Mesentery + + ++ ++ 

Histiocytic sarcoma X

Pancreas +++++++++++++++++ 

Histiocytic sarcoma X

Salivary glands +++++++++++++++++ 

Stomach, forestomach +++++++++++++++++ 

Squamous cell papilloma X X

Stomach, glandular +++++++++++++++++ 

Cardiovascular System

Blood vessel +++++++++++++++++ 

Heart +++++++++++++++++ 

Histiocytic sarcoma X

Endocrine System

Adrenal cortex +++++++++++++++++ 

Adrenal medulla +++++++++++++++++ 

Islets, pancreatic +++++++++++++++++ 

Parathyroid gland + M ++++++ M + M ++++ M M

Pituitary gland +++++++++++++++++ 

Meningioma malignant, metastatic, brain X

Pars distalis, adenoma XXX

Thyroid gland +++++++++++++++++ M ++++++ 

Follicular cell, adenoma

General Body System

None

Genital System

Clitoral gland ++++++++ M +++++ M ++ 

Ovary +++++++++++++++++ 

Cystadenoma X

Histiocytic sarcoma X

Uterus +++++++++++++++++ 

Histiocytic sarcoma X X

Polyp stromal X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 175 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0000000011111111112222222 

Carcass ID Number

222

677

946

222

889

051

2222222222

9955566678

2346806824

232222222 

807778999 T

803591789 T

Total

issues/ 

umors

Alimentary System

Esophagus 16 

Gallbladder 17 

Intestine large, colon 17 

Intestine large, rectum 17 

Intestine large, cecum 16 

Intestine small, duodenum 17 

Intestine small, jejunum 16 

Intestine small, ileum 17 

Liver +++++++++++++++++++++++++ 49 

Hemangiosarcoma X 1 

Hepatocellular carcinoma X X X 10 

Hepatocellular carcinoma, multiple X X X 3 

Hepatocellular adenoma X X X X X X X 8 

Hepatocellular adenoma, multiple X X X X 9 

Histiocytic sarcoma 2 

Mesentery 6 

Histiocytic sarcoma 1 

Pancreas 17 

Histiocytic sarcoma 1 

Salivary glands 17 

Stomach, forestomach 17 

Squamous cell papilloma 2 

Stomach, glandular 17 

Cardiovascular System

Blood vessel 17 

Heart 17 

Histiocytic sarcoma 1 

Endocrine System

Adrenal cortex 17 

Adrenal medulla 17 

Islets, pancreatic 17 

Parathyroid gland 12 

Pituitary gland 17 

Meningioma malignant, metastatic, brain 1 

Pars distalis, adenoma 3 

Thyroid gland +++++++++++++++++++++++++ 48 

Follicular cell, adenoma X X X X X 5 

General Body System

None

Genital System

Clitoral gland 15 

Ovary 17 

Cystadenoma 1 

Histiocytic sarcoma 1 

Uterus 17 

Histiocytic sarcoma 2 

Polyp stromal 1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

176 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

(continued) 

034445666666677777777777 

Number of Days on Study 231463023456711334444444 

471678456296428150000000 

222222222222222222222222 

Carcass ID Number 788597785987598965556666 

169550777038242651391234 

Hematopoietic System

Bone marrow +++++++++++++++++ 

Histiocytic sarcoma

Lymph node

Lumbar, histiocytic sarcoma

Mediastinal, histiocytic sarcoma

Lymph node, mandibular

Carcinoma, metastatic, harderian gland

Lymph node, mesenteric

Histiocytic sarcoma

Spleen

Hemangiosarcoma

Thymus

X X

+ ++ 

X X

X

++++++++++++ M + + M + 

X

+++++++++++++++++ 

X X

+++++++++++++++++ 

X

+++++++++ M +++++++ 

Integumentary System

Mammary gland

Skin

+++++++++++++++++ 

++++++++++++++++++++++++ 

Subcutaneous tissue, skin, site of

application, fibrosarcoma X

Musculoskeletal System

Bone +++++++++++++++++ 

Nervous System

Brain +++++++++++++++++ 

Carcinoma, metastatic, harderian gland

Meningioma malignant

Peripheral nerve

Spinal cord

X

+ 

+ 

X

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Carcinoma, metastatic, harderian gland

Hepatocellular carcinoma, metastatic, liver

Histiocytic sarcoma

Nose

+++++++++++++++++ 

X

X X

X

X

+++++++++++++++++ 

Trachea +++++++++++++++++ 

Special Senses System

Eye

Carcinoma, metastatic, harderian gland

Harderian gland

Carcinoma

+ + 

X

+

X

+ 

X

Urinary System

Kidney

Histiocytic sarcoma

Urinary bladder

+++++++++++++++++ 

X X

+++++++++++++++++ 

Systemic Lesions

Multiple organs ++++++++++++++++++++++++ 

Histiocytic sarcoma X X

Lymphoma malignant X X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 177 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 182 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0000000011111111112222222 

Carcass ID Number

2222222222222222232222222 

6778899955566678807778999 T

9460512346806824803591789 T

Total

issues/ 

umors

Hematopoietic System

Bone marrow 17 

Histiocytic sarcoma 2 

Lymph node 3 

Lumbar, histiocytic sarcoma 2 

Mediastinal, histiocytic sarcoma 1 

Lymph node, mandibular 15 

Carcinoma, metastatic, harderian gland 1 

Lymph node, mesenteric 17 

Histiocytic sarcoma 2 

Spleen 17 

Hemangiosarcoma 1 

Thymus 16 

Integumentary System

Mammary gland 17 

Skin +++++++++++++++++++++++++ 49 

Subcutaneous tissue, skin, site of

application, fibrosarcoma 1 

Musculoskeletal System

Bone 17 

Nervous System

Brain

Carcinoma, metastatic, harderian gland

Meningioma malignant

Peripheral nerve

Spinal cord

17 

1 

1 

1 

1 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Carcinoma, metastatic, harderian gland

Hepatocellular carcinoma, metastatic, liver

Histiocytic sarcoma

Nose

Trachea

+ 

X

18 

1 

2 

2 

1 

17 

17 

Special Senses System

Eye

Carcinoma, metastatic, harderian gland

Harderian gland

Carcinoma

2 

1 

2 

2 

Urinary System

Kidney 17 

Histiocytic sarcoma 2 

Urinary bladder 17 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 49 

Histiocytic sarcoma 2 

Lymphoma malignant 3 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

178 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

5555666666666666777777777 

Number of Days on Study 6678002345567778234444444 

1258122624912495820000000 

3333333333333333333333333 

Carcass ID Number 4121323044030011110111234 

1652746363216714309578392 

Alimentary System

Esophagus ++++++++++++++++++ 

Gallbladder ++++++++++++++++++ 

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Intestine large, colon ++++++++++++++++++ 

Intestine large, rectum ++++++++++++++++++ 

Intestine large, cecum ++++++++++++++++++ 

Leiomyosarcoma X

Intestine small, duodenum ++++++++++++++++++ 

Intestine small, jejunum ++++++++++++++++++ 

Leiomyosarcoma X

Intestine small, ileum ++++++++++++++++++ 

Liver +++++++++++++++++++++++++ 

Hemangiosarcoma

Hepatocellular carcinoma X X X X X X

Hepatocellular adenoma X X X X X

Hepatocellular adenoma, multiple X X

Hepatocholangiocarcinoma X

Histiocytic sarcoma X

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Mesentery + + + ++ 

Hepatocholangiocarcinoma, metastatic, 

liver X

Histiocytic sarcoma X

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Pancreas ++++++++++++++++++ 

Hepatocholangiocarcinoma, metastatic, 

liver X

Histiocytic sarcoma X

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Salivary glands ++++++++++++++++++ 

Stomach, forestomach ++++++++++++++++++ 

Stomach, glandular ++++++++++++++++++ 

Carcinoma X

Hepatocholangiocarcinoma, metastatic, 

liver X

Cardiovascular System

Blood vessel ++++++++++++++++++ 

Aorta, hepatocholangiocarcinoma, 

metastatic, liver X

Heart ++++++++++++++++++ 

Alveolar/bronchiolar carcinoma, 

metastatic, lung X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 179 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0011111111111112222222222 

3333333333333333333333333 Total

Carcass ID Number 4400122233334450022223344 Tissues/ 

4845908923457901812670805 Tumors

Alimentary System

Esophagus 18 

Gallbladder 18 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Intestine large, colon 18 

Intestine large, rectum 18 

Intestine large, cecum 18 

Leiomyosarcoma 1 

Intestine small, duodenum 18 

Intestine small, jejunum 18 

Leiomyosarcoma 1 

Intestine small, ileum 18 

Liver + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hemangiosarcoma X 1 

Hepatocellular carcinoma X 7 

Hepatocellular adenoma X X X X X XXX X 14 

Hepatocellular adenoma, multiple X X 4 

Hepatocholangiocarcinoma 1 

Histiocytic sarcoma X 2 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Mesentery 5 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Histiocytic sarcoma 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Pancreas 18 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Histiocytic sarcoma 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Salivary glands 18 

Stomach, forestomach 18 

Stomach, glandular 18 

Carcinoma 1 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Cardiovascular System

Blood vessel

Aorta, hepatocholangiocarcinoma, 

metastatic, liver

Heart

Alveolar/bronchiolar carcinoma, 

metastatic, lung

18 

1 

18 

1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

180 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

5555666666666666777777777 

Number of Days on Study 6678002345567778234444444 

1258122624912495820000000 

Carcass ID Number

3333333333333333333333333 

4121323044030011110111234 

1652746363216714309578392 

Endocrine System

Adrenal cortex ++++++++++++++++++ 

Hepatocholangiocarcinoma, metastatic, 

liver X

Adrenal medulla ++++++++++++++++++ 

Islets, pancreatic ++++++++++++++++++ 

Adenoma X

Parathyroid gland M M +++ M M + + M M ++++ M ++ 

Pituitary gland ++++++++++++++++++ 

Pars distalis, adenoma X

Thyroid gland +++++++++++++++++++++++++ 

Follicular cell, adenoma X

General Body System

None

Genital System

Clitoral gland + + M ++++ M +++ M ++++++ 

Ovary ++++++++++++++++++ 

Hemangioma X

Hepatocholangiocarcinoma, metastatic, 

liver X

Histiocytic sarcoma X

Uterus ++++++++++++++++++ 

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Hematopoietic System

Bone marrow ++++++++++++++++++ 

Lymph node + ++ + ++ 

Bronchial, alveolar/bronchiolar

carcinoma, metastatic, lung X

Inguinal, histiocytic sarcoma X

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver X

Pancreatic, histiocytic sarcoma X

Renal, histiocytic sarcoma X

Lymph node, mandibular +++++++++++ M M ++++ M

Lymph node, mesenteric + M ++++++++++++++++ 

Hepatocholangiocarcinoma, metastatic, 

liver X

Histiocytic sarcoma X

Leiomyosarcoma, metastatic, 

intestine small, jejunum X

Spleen ++++++++++++++++++ 

Histiocytic sarcoma X

Thymus ++++++++++++++++ M M

Alveolar/bronchiolar carcinoma, 

metastatic, lung X

Hepatocholangiocarcinoma, metastatic, 

liver X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 181 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0011111111111112222222222 

3333333333333333333333333 Total

Carcass ID Number 4400122233334450022223344 Tissues/ 

4845908923457901812670805 Tumors

Endocrine System

Adrenal cortex 18 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Adrenal medulla 18 

Islets, pancreatic 18 

Adenoma 1 

Parathyroid gland 11 

Pituitary gland 18 

Pars distalis, adenoma 1 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Follicular cell, adenoma 1 

General Body System

None

Genital System

Clitoral gland

Ovary

Hemangioma

Hepatocholangiocarcinoma, metastatic, 

liver

Histiocytic sarcoma

Uterus

Leiomyosarcoma, metastatic, 

intestine small, jejunum

15 

18 

1 

1 

1 

18 

1 

Hematopoietic System

Bone marrow

Lymph node

Bronchial, alveolar/bronchiolar

carcinoma, metastatic, lung

Inguinal, histiocytic sarcoma

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver

Pancreatic, histiocytic sarcoma

Renal, histiocytic sarcoma

Lymph node, mandibular

Lymph node, mesenteric

Hepatocholangiocarcinoma, metastatic, 

liver

Histiocytic sarcoma

Leiomyosarcoma, metastatic, 

intestine small, jejunum

Spleen

Histiocytic sarcoma

Thymus

Alveolar/bronchiolar carcinoma, 

metastatic, lung

Hepatocholangiocarcinoma, metastatic, 

liver

18 

6 

1 

1 

1 

1 

1 

15 

17 

1 

1 

1 

18 

1 

16 

1 

1 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

182 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

Number of Days on Study

5555666666666666777777777 

6678002345567778234444444 

1258122624912495820000000 

3333333333333333333333333 

Carcass ID Number 4121323044030011110111234 

1652746363216714309578392 

Integumentary System

Mammary gland

Skin

Subcutaneous tissue, skin, site of

application, fibrosarcoma

M +++++++++++ M +++++ 

+++++++++++++++++++++++++ 

Musculoskeletal System

Bone

Skeletal muscle

Hepatocholangiocarcinoma, metastatic, 

liver

++++++++++++++++++ 

+ 

X

Nervous System

Brain ++++++++++++++++++ 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Hepatocellular carcinoma, metastatic, 

liver

Hepatocholangiocarcinoma, metastatic, 

liver

Leiomyosarcoma, metastatic, 

intestine small, jejunum

Leiomyosarcoma, metastatic, 

intestine large, cecum

Mediastinum, alveolar/bronchiolar

carcinoma, metastatic, lung

Nose

Trachea

++++++++++++++++++ 

X X

X

X

X

X

X

X

++++++++++++++++++ 

++++++++++++++++++ 

Special Senses System

Harderian gland

Carcinoma

+ 

X

Urinary System

Kidney

Urinary bladder

++++++++++++++++++ 

++++++++++++++++++ 

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Lymphoma malignant

+++++++++++++++++++++++++ 

X

X X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 183 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 364 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

0011111111111112222222222 

3333333333333333333333333 Total

Carcass ID Number 4400122233334450022223344 Tissues/ 

4845908923457901812670805 Tumors

Integumentary System

Mammary gland 16 

Skin +++++++++++++++++++++++++ 50 

Subcutaneous tissue, skin, site of

application, fibrosarcoma X 1 

Musculoskeletal System

Bone 18 

Skeletal muscle 1 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Nervous System

Brain 18 

Respiratory System

Lung 18 

Alveolar/bronchiolar adenoma 2 

Alveolar/bronchiolar carcinoma 1 

Hepatocellular carcinoma, metastatic, 

liver 1 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Leiomyosarcoma, metastatic, 

intestine small, jejunum 1 

Leiomyosarcoma, metastatic, 

intestine large, cecum 1 

Mediastinum, alveolar/bronchiolar

carcinoma, metastatic, lung 1 

Nose 18 

Trachea 18 

Special Senses System

Harderian gland

Carcinoma

1 

1 

Urinary System

Kidney

Urinary bladder

18 

18 

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Lymphoma malignant

+++++++++++++++++++++++++ 

X

50 

2 

3 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

184 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

Number of Days on Study

0456666666777777777777777 

7031227899013344444444444 

2342795614770800000000001 

Carcass ID Number

3333333333333333333333333 

9566868697957955567788895 

5460041502661112786826833 

Alimentary System

Esophagus

Gallbladder

Intestine large, colon

Intestine large, rectum

Intestine large, cecum

Intestine small, duodenum

Intestine small, jejunum

Fibrosarcoma, metastatic, skin

Intestine small, ileum

Liver

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Hepatocholangiocarcinoma

Histiocytic sarcoma

Mesentery

Fibrosarcoma, metastatic, skin

Hemangioma

Hepatocholangiocarcinoma, metastatic, 

liver

Pancreas

Fibrosarcoma, metastatic, skin

Salivary glands

Stomach, forestomach

Stomach, glandular

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X X X X

X X

X X X X X X X X

X X

X X

X X

 +++ ++ + + 

X

X

X X

+++++++++++++++++++++++++ 

X

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

Cardiovascular System

Blood vessel +++++++++++++++++++++++++ 

Heart +++++++++++++++++++++++++ 

Endocrine System

Adrenal cortex +++++++++++++++++++++++++ 

Capsule, adenoma X

Adrenal medulla + M +++++++++++++++++++ M +++ 

Pheochromocytoma benign X

Islets, pancreatic +++++++++++++++++++++++++ 

Parathyroid gland M + M + + M +++++ M M ++++++++++ M + 

Pituitary gland +++++++++++++++++++++++++ 

Pars distalis, adenoma X X

Pars intermedia, adenoma X

Pars intermedia, carcinoma X

Thyroid gland +++++++++++++++++++++++++ 

Follicular cell, adenoma X X

Follicular cell, carcinoma

General Body System

None
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 185 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

Number of Days on Study

7777777777777777777777777 

4444444444444444444444444 

1111111111112222222222222 

Carcass ID Number

3333333333343333333333333 

5666678889905567777788999 

9137954572705820347939489 

Total

Tissues/ 

Tumors

Alimentary System

Esophagus

Gallbladder

Intestine large, colon

Intestine large, rectum

Intestine large, cecum

Intestine small, duodenum

Intestine small, jejunum

Fibrosarcoma, metastatic, skin

Intestine small, ileum

Liver

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Hepatocholangiocarcinoma

Histiocytic sarcoma

Mesentery

Fibrosarcoma, metastatic, skin

Hemangioma

Hepatocholangiocarcinoma, metastatic, 

liver

Pancreas

Fibrosarcoma, metastatic, skin

Salivary glands

Stomach, forestomach

Stomach, glandular

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

X X X X

X X X X X X XXX X X

X X X X X X X

+++ + + 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

+++++++++++++++++++++++++ 

50 

50 

50 

50 

50 

50 

50 

1 

50 

50 

8 

2 

19 

9 

2 

2 

12 

1 

1 

2 

50 

1 

50 

50 

50 

Cardiovascular System

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Heart + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Endocrine System

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Capsule, adenoma 1 

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + + 48 

Pheochromocytoma benign 1 

Islets, pancreatic + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Parathyroid gland M + + M + M + M ++++ M + M M ++++++ M + + 36 

Pituitary gland +++++++ M + + + + + + + + + + + + + + + + + 49 

Pars distalis, adenoma X X X 5 

Pars intermedia, adenoma X 2 

Pars intermedia, carcinoma 1 

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Follicular cell, adenoma X X X X 6 

Follicular cell, carcinoma X 1 

General Body System

None
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

186 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

0456666666777777777777777 

Number of Days on Study 7031227899013344444444444 

2342795614770800000000001 

3333333333333333333333333 

Carcass ID Number 9566868697957955567788895 

5460041502661112786826833 

Genital System

Clitoral gland +++++++++++++ M + M +++++++++ 

Hepatocholangiocarcinoma, metastatic, 

liver X

Ovary + + M ++++++++++++++++++++++ 

Cystadenoma

Fibrosarcoma, metastatic, skin X

Histiocytic sarcoma X

Oviduct

Uterus +++++++++++++++++++++++++ 

Hemangiosarcoma

Histiocytic sarcoma X

Hematopoietic System

Bone marrow +++++++++++++++++++++++++ 

Lymph node + + + + + + 

Lumbar, histiocytic sarcoma X X

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver X

Mediastinal, histiocytic sarcoma X

Renal, hepatocholangiocarcinoma, 

metastatic, liver X

Renal, histiocytic sarcoma X

Lymph node, mandibular ++++ M ++++++++++++++++++++ 

Histiocytic sarcoma X

Lymph node, mesenteric +++++++++++++++++++++++++ 

Fibrosarcoma, metastatic, skin X

Hepatocholangiocarcinoma, metastatic, 

liver X X

Histiocytic sarcoma X X

Spleen +++++++++++++++++++++++++ 

Hemangiosarcoma X

Histiocytic sarcoma X

Thymus +++++++ M + M + M +++++++++++++ 

Hepatocholangiocarcinoma, metastatic, 

liver X X

Histiocytic sarcoma X

Integumentary System

Mammary gland +++++++++++++++++++++++++ 

Carcinoma

Skin +++++++++++++++++++++++++ 

Subcutaneous tissue, fibrosarcoma X

Subcutaneous tissue, hemangiosarcoma

Musculoskeletal System

Bone +++++++++++++++++++++++++ 

Skeletal muscle + + + 

Fibrosarcoma, metastatic, skin X

Hepatocholangiocarcinoma, metastatic, 

liver X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 187 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

1111111111112222222222222 

3333333333343333333333333 Total

Carcass ID Number 5666678889905567777788999 Tissues/ 

9137954572705820347939489 Tumors

Genital System

Clitoral gland ++++++++++++++++++++ M + M + + 46 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Ovary + + + + + + + + + + + + + + + + + + + + + + + + + 49 

Cystadenoma X 1 

Fibrosarcoma, metastatic, skin 1 

Histiocytic sarcoma 1 

Oviduct + 1 

Uterus + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hemangiosarcoma X 1 

Histiocytic sarcoma 1 

Hematopoietic System

Bone marrow + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Lymph node 6 

Lumbar, histiocytic sarcoma 2 

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver 1 

Mediastinal, histiocytic sarcoma 1 

Renal, hepatocholangiocarcinoma, 

metastatic, liver 1 

Renal, histiocytic sarcoma 1 

Lymph node, mandibular +++++++++ M + + + + + + + + + + + + + + + 48 

Histiocytic sarcoma 1 

Lymph node, mesenteric + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Fibrosarcoma, metastatic, skin 1 

Hepatocholangiocarcinoma, metastatic, 

liver 2 

Histiocytic sarcoma 2 

Spleen + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Hemangiosarcoma X 2 

Histiocytic sarcoma 1 

Thymus + + M +++++++++++++++++ M + M + + 44 

Hepatocholangiocarcinoma, metastatic, 

liver 2 

Histiocytic sarcoma 1 

Integumentary System

Mammary gland + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Carcinoma X 1 

Skin + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Subcutaneous tissue, fibrosarcoma 1 

Subcutaneous tissue, hemangiosarcoma X 1 

Musculoskeletal System

Bone + + + + + + + + + + + + + + + + + + + + + + + + + 50 

Skeletal muscle 3 

Fibrosarcoma, metastatic, skin 1 

Hepatocholangiocarcinoma, metastatic, 

liver 2 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

188 Sodium Xylenesulfonate, NTP TR 464 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

0456666666777777777777777 

Number of Days on Study 7031227899013344444444444 

2342795614770800000000001 

3333333333333333333333333 

Carcass ID Number 9566868697957955567788895 

5460041502661112786826833 

Nervous System

Brain +++++++++++++++++++++++++ 

Peripheral nerve

Spinal cord

+ 

+ 

Respiratory System

Lung

Alveolar/bronchiolar adenoma

Carcinoma, metastatic, harderian

gland

Hepatocellular carcinoma, metastatic, 

liver

+++++++++++++++++++++++++ 

X X

X

X

Hepatocholangiocarcinoma, metastatic, 

liver X X

Histiocytic sarcoma

Squamous cell carcinoma, metastatic, 

urinary bladder

Nose

X X

X

+++++++++++++++++++++++++ 

Histiocytic sarcoma

Trachea

X

+++++++++++++++++++++++++ 

Special Senses System

Harderian gland

Adenoma

+

X

+ + 

Carcinoma X X

Urinary System

Kidney

Hepatocholangiocarcinoma, metastatic, 

liver

+++++++++++++++++++++++++ 

X

Urinary bladder

Squamous cell carcinoma

+++++++++++++++++++++++++ 

X

Systemic Lesions

Multiple organs

Histiocytic sarcoma

Lymphoma malignant

+++++++++++++++++++++++++ 

X X

X X X
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 189 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate: 727 mg/kg

(continued) 

7777777777777777777777777 

Number of Days on Study 4444444444444444444444444 

1111111111112222222222222 

3333333333343333333333333 Total

Carcass ID Number 5666678889905567777788999 Tissues/ 

9137954572705820347939489 Tumors

Nervous System

Brain +++++++++++++++++++++++++ 50 

Peripheral nerve 1 

Spinal cord 1 

Respiratory System

Lung +++++++++++++++++++++++++ 50 

Alveolar/bronchiolar adenoma X X X X 6 

Carcinoma, metastatic, harderian

gland 1 

Hepatocellular carcinoma, metastatic, 

liver X X 3 

Hepatocholangiocarcinoma, metastatic, 

liver 2 

Histiocytic sarcoma 2 

Squamous cell carcinoma, metastatic, 

urinary bladder 1 

Nose +++++++++++++++++++++++++ 50 

Histiocytic sarcoma 1 

Trachea +++++++++++++++++++++++++ 50 

Special Senses System

Harderian gland 3 

Adenoma 1 

Carcinoma 2 

Urinary System

Kidney +++++++++++++++++++++++++ 50 

Hepatocholangiocarcinoma, metastatic, 

liver 1 

Urinary bladder +++++++++++++++++++++++++ 50 

Squamous cell carcinoma 1 

Systemic Lesions

Multiple organs +++++++++++++++++++++++++ 50 

Histiocytic sarcoma 2 

Lymphoma malignant X X 5 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

190 Sodium Xylenesulfonate, NTP TR 464 

TABLE D3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Harderian Gland: Adenoma

Overall ratea

Adjusted rateb

Terminal ratec

First incidence (days) 

Life table testd

Logistic regression testd

Fisher exact testd

3/50 (6%) 

8.9% 

2/31 (6%) 

670 

0/49 (0%) e 0/50 (0%) e 1/50 (2%) 

2.5% 

0/36 (0%) 

707 

P ̆0.258N

P ̆0.292N

P ̆0.309N

Harderian Gland: Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

4/50 (8%) 

10.7% 

2/31 (6%) 

453 

2/49 (4%) e 1/50 (2%) e 3/50 (6%) 

7.9% 

2/36 (6%) 

707 

P ̆0.435N

P ̆0.506N

P ̆0.500N

Liver: Hepatocellular Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage testd

Fisher exact test

18/50 (36%) 

54.1% 

16/31 (52%) 

621 

P ̆0.060 

P ̆0.034 

P ̆0.016 

17/49 (35%) 

47.8% 

14/32 (44%) 

446 

P ̆0.459N

P ̆0.533N

P ̆0.530N

18/50 (36%) 

49.0% 

14/32 (44%) 

601 

P ̆0.540N

P ̆0.553N

P ̆0.582N

28/50 (56%) 

69.8% 

24/36 (67%) 

686 

P ̆0.118 

P ̆0.080 

P ̆0.035 

Liver: Hepatocellular Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

10/50 (20%) 

25.5% 

4/31 (13%) 

495 

P ̆0.292N

P ̆0.397N

P ̆0.399N

13/49 (27%) 

36.3% 

10/32 (31%) 

411 

P ̆0.342 

P ̆0.293 

P ̆0.298 

7/50 (14%) 

17.0% 

2/32 (6%) 

575 

P ̆0.297N

P ̆0.360N

P ̆0.298N

10/50 (20%) 

24.7% 

6/36 (17%) 

675 

P ̆0.469N

P ̆0.595N

P ̆0.598N

Liver: Hepatocellular Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Cochran-Armitage test

Fisher exact test

27/50 (54%) 

68.6% 

19/31 (61%) 

495 

P ̆0.295 

P ̆0.131 

P ̆0.088 

23/49 (47%) 

61.4% 

18/32 (56%) 

411 

P ̆0.254N

P ̆0.326N

P ̆0.308N

23/50 (46%) 

56.4% 

15/32 (47%) 

575 

P ̆0.259N

P ̆0.237N

P ̆0.274N

33/50 (66%) 

76.7% 

26/36 (72%) 

675 

P ̆0.448 

P ̆0.237 

P ̆0.154 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 191 

TABLE D3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

(continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Lung: Alveolar/bronchiolar Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

4/50 (8%) 

11.5% 

2/31 (6%) 

659 

1/18 (6%) e 2/18 (11%) e 6/50 (12%) 

16.7% 

6/36 (17%) 

740 (T) 

P ̆0.461 

P ̆0.433 

P ̆0.370 

Lung: Alveolar/bronchiolar Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

2/50 (4%) 

6.5% 

2/31 (6%) 

740 (T) 

0/18 (0%) e 1/18 (6%) e 0/50 (0%) 

0.0% 

0/36 (0%) 

—f

P ̆0.206N

P ̆0.206N

P ̆0.247N

Lung: Alveolar/bronchiolar Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

5/50 (10%) 

14.6% 

3/31 (10%) 

659 

1/18 (6%) e 3/18 (17%) e 6/50 (12%) 

16.7% 

6/36 (17%) 

740 (T) 

P ̆0.598 

P ̆0.574 

P ̆0.500 

Pancreatic Islets: Adenoma or Carcinoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

2/50 (4%) 

4.3% 

0/31 (0%) 

495 

0/17 (0%) e 1/18 (6%) e 0/50 (0%) 

0.0% 

0/36 (0%) 

—

P ̆0.233N

P ̆0.204N

P ̆0.247N

Pituitary Gland (Pars Distalis): Adenoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

7/50 (14%) 

20.4% 

5/31 (16%) 

621 

3/17 (18%) e 1/18 (6%) e 5/49 (10%) 

12.9% 

3/35 (9%) 

686 

P ̆0.299N

P ̆0.357N

P ̆0.394N
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

192 Sodium Xylenesulfonate, NTP TR 464 

TABLE D3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

(continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Pituitary Gland (Pars Intermedia): Adenoma or Carcinoma e e Overall rate 1/50 (2%) 0/17 (0%) 0/18 (0%) 3/49 (6%) 

Adjusted rate 3.2% 7.8% 

Terminal rate 1/31 (3%) 2/35 (6%) 

First incidence (days) 740 (T) 629 

Life table test P ̆0.348 

Logistic regression test P ̆0.307 

Fisher exact test P ̆0.301 

Spleen: Hemangiosarcoma e e Overall rate 0/50 (0%) 1/17 (6%) 0/18 (0%) 2/50 (4%) 

Adjusted rate 0.0% 5.6% 

Terminal rate 0/31 (0%) 2/36 (6%) 

First incidence (days) — 740 (T) 

Life table test P ̆0.272 

Logistic regression test P ̆0.272 

Fisher exact test P ̆0.247 

Thyroid Gland (Follicular Cell): Adenoma

Overall rate 2/50 (4%) 5/48 (10%) 1/50 (2%) 6/50 (12%) 

Adjusted rate 5.5% 16.1% 3.1% 16.0% 

Terminal rate 1/31 (3%) 5/31 (16%) 1/32 (3%) 5/36 (14%) 

First incidence (days) 621 740 (T) 740 (T) 707 

Life table test P ̆0.214 P ̆0.215 P ̆0.490N P ̆0.187 

Logistic regression test P ̆0.197 P ̆0.197 P ̆0.498N P ̆0.151 

Cochran-Armitage test P ̆0.157 

Fisher exact test P ̆0.201 P ̆0.500N P ̆0.134 

Thyroid Gland (Follicular Cell): Adenoma or Carcinoma

Overall rate 2/50 (4%) 5/48 (10%) 1/50 (2%) 7/50 (14%) 

Adjusted rate 5.5% 16.1% 3.1% 18.8% 

Terminal rate 1/31 (3%) 5/31 (16%) 1/32 (3%) 6/36 (17%) 

First incidence (days) 621 740 (T) 740 (T) 707 

Life table test P ̆0.124 P ̆0.215 P ̆0.490N P ̆0.122 

Logistic regression test P ̆0.112 P ̆0.197 P ̆0.498N P ̆0.096 

Cochran-Armitage test P ̆0.084 

Fisher exact test P ̆0.201 P ̆0.500N P ̆0.080 

Uterus: Stromal Polyp e e Overall rate 3/50 (6%) 1/17 (6%) 0/18 (0%) 0/50 (0%)
#

Adjusted rate 9.4% 0.0%
#

Terminal rate 2/31 (6%) 0/36 (0%)
#

First incidence (days) 734 —
 

Life table test P ̆0.098N
 

Logistic regression test P ̆0.096N
 

Fisher exact test P ̆0.121N
 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 193 

TABLE D3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

(continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

All Organs: Hemangiosarcoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

0/50 (0%) 

0.0% 

0/31 (0%) 

—

2/49 (4%) e 1/50 (2%) e 4/50 (8%) 

11.1% 

4/36 (11%) 

740 (T) 

P ̆0.083 

P ̆0.083 

P ̆0.059 

All Organs: Hemangioma or Hemangiosarcoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

2/50 (4%) 

5.7% 

1/31 (3%) 

670 

2/49 (4%) e 2/50 (4%) e 5/50 (10%) 

13.4% 

4/36 (11%) 

717 

P ̆0.281 

P ̆0.249 

P ̆0.218 

All Organs: Histiocytic Sarcoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

3/50 (6%) 

7.0% 

0/31 (0%) 

499 

2/49 (4%) e 2/50 (4%) e 2/50 (4%) 

5.4% 

1/36 (3%) 

738 

P ̆0.450N

P ̆0.507N

P ̆0.500N

All Organs: Malignant Lymphoma

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

8/50 (16%) 

24.7% 

7/31 (23%) 

692 

3/17 (18%) e 3/18 (17%) e 5/50 (10%) 

11.7% 

2/36 (6%) 

403 

P ̆0.204N

P ̆0.275N

P ̆0.277N

All Organs: Benign Neoplasms

Overall rate

Adjusted rate

Terminal rate

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

29/50 (58%) 

74.1% 

21/31 (68%) 

589 

22/49 (45%)e 20/50 (40%)e 35/50 (70%) 

83.3% 

29/36 (81%) 

686 

P ̆0.471 

P ̆0.303 

P ̆0.149 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

194 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE D3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-Year Dermal Study of Sodium Xylenesulfonate

(continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

All Organs: Malignant Neoplasms

Overall rate 25/50 (50%) 22/49 (45%)e 18/50 (36%)e 26/50 (52%) 

Adjusted rate 57.3% 54.0% 

Terminal rate 13/31 (42%) 14/36 (39%) 

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

453 403 

P ̆0.421N

P ̆0.331 

P ̆0.500 

All Organs: Benign or Malignant Neoplasms

Overall rate 45/50 (90%) 34/49 (69%)e 31/50 (62%)e 44/50 (88%) 

Adjusted rate 93.7% 89.8% 

Terminal rate 28/31 (90%) 31/36 (86%) 

First incidence (days) 

Life table test

Logistic regression test

Fisher exact test

453 403 

P ̆0.164N

P ̆0.465N

P ̆0.500N

(T)Terminal sacrifice a Number of neoplasm-bearing animals/number of animals examined. Denominator is number of animals examined microscopically for liver, lung, pancreatic

islets, pituitary gland, spleen, 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 dosed group incidence are the P values corresponding to pairwise

comparisons between the controls and that dosed 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 a dose group is indicated by N. e Tissues (except skin) were examined microscopically only in those animals dying prior to terminal sacrifice or when it was observed to be abnormal at
 

necropsy; thus statistical comparison with the controls are not applicable.
#

f	 Not applicable; no neoplasms in animal group
 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 195 

TABLE D4 

Historical a Incidence of Hepatocellular Neoplasms in Control Female B6C3F1 Mice

Incidence in Controls

Study Adenoma Carcinoma Adenoma

or Carcinoma

Historical Incidence at Battelle Columbus: Dermal (Acetone) Studies

4-Vinyl-1-cyclohexene Diepoxide 8/50 2/50 10/50 

Triethanolamine 22/50 1/50 23/50 

Overall Historical Incidence: Dermal (Acetone) Studies

Total 34/150 (22.7%) 7/150 (4.7%) 40/150 (26.7%) 

Standard deviation 18.9% 3.1% 17.0% 

Range 8%-44% 2%-8% 14%-46% 

Historical Incidence at Battelle Columbus: Dermal (Ethanol) Study

Benzethonium Chloride 20/52 (38.5%) 12/52 (23.1%) 27/52 (51.9%) 

Historical Incidence at Battelle Columbus: Feed Studies

4,4-Thiobis(6-t-butyl-m-cresol) 17/51 4/51 20/51 

5,5-Diphenylhydantoin 5/48 0/48 5/48 

Pentachlorophenol (Dowicide EC-7) 1/34 0/34 1/34 

Ethylene Thiourea 2/50 2/50 4/50 

Polybrominated Biphenyls (Firemaster FF-1®) 4/50 1/50 5/50 

Manganese (II) Sulfate Monohydrate 12/51 3/51 13/51 

Oxazepam 25/50 9/50 28/50 

Technical Grade Pentachlorophenol 3/33 0/33 3/33 

Triamterene 10/50 4/50 13/50 

Triamterene 7/50 5/50 10/50 

Tricresyl Phosphate 12/50 10/50 21/50 

Overall Historical Incidence: Feed Studies

Total 231/1,464 (15.8%) 108/1,464 (7.4%) 313/1,464 (21.4%) 

Standard deviation 10.6% 6.1% 13.0% 

Range 2%-50% 0%-20% 3%-56% 

a Data as of 12 May 1995 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

196 Sodium Xylenesulfonate, NTP TR 464 

TABLE D5 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Disposition Summary

Animals initially in study 50 50 50 50 

Early deaths

Moribund 12 7 9 6 

Natural deaths 7 10 9 8 

Survivors

Terminal sacrifice 31 32 32 36 

Missing 1 

Animals examined microscopically 50 49 50 50 

Alimentary System

Esophagus (50) (16) (18) (50) 

Inflammation 1 (6%) 

Intestine small, duodenum (50) (17) (18) (50) 

Inflammation 1 (2%) 

Intestine small, ileum (50) (17) (18) (50) 

Hyperplasia, lymphoid 1 (2%) 

Liver (50) (49) (50) (50) 

Angiectasis 1 (2%) 

Basophilic focus 3 (6%) 2 (4%) 1 (2%) 

Clear cell focus 1 (2%) 

Eosinophilic focus 14 (28%) 11 (22%) 11 (22%) 10 (20%) 

Hematopoietic cell proliferation 2 (4%) 2 (4%) 1 (2%) 1 (2%) 

Infarct 1 (2%) 

Inflammation, chronic active 22 (44%) 28 (57%) 24 (48%) 21 (42%) 

Mixed cell focus 10 (20%) 3 (6%) 1 (2%) 7 (14%) 

Necrosis 3 (6%) 4 (8%) 9 (18%) 3 (6%) 

Pigmentation, hemosiderin 1 (2%) 1 (2%) 

Bile duct, cyst 2 (4%) 

Bile duct, hyperplasia 4 (8%) 2 (4%) 4 (8%) 2 (4%) 

Serosa, necrosis 1 (2%) 

Mesentery (7) (6) (5) (12) 

Inflammation 1 (14%) 1 (20%) 

Fat, necrosis 3 (43%) 2 (33%) 7 (58%) 

Pancreas (50) (17) (18) (50) 

Atrophy 3 (6%) 1 (6%) 6 (12%) 

Cytoplasmic alteration 1 (2%) 1 (2%) 

Inflammation 1 (2%) 

Necrosis 1 (2%) 

Duct, cyst 2 (4%) 2 (12%) 1 (6%) 5 (10%) 

Salivary glands (50) (17) (18) (50) 

Atrophy 1 (2%) 1 (2%) 

Fibrosis 1 (2%) 

Stomach, forestomach (50) (17) (18) (50) 

Erosion 1 (6%) 

Hyperplasia, focal 1 (2%) 1 (6%) 3 (6%) 

a Number of animals examined microscopically at the site and the number of animals with lesion
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 197 

TABLE D5 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Cardiovascular System

Blood vessel (50) (17) (18) (50) 

Aorta, inflammation, chronic active 1 (2%) 

Heart (50) (17) (18) (50) 

Degeneration 2 (12%) 

Mineralization 5 (10%) 1 (2%) 

Atrium, thrombosis 1 (2%) 

Epicardium, inflammation, suppurative 1 (6%) 

Endocrine System

Adrenal cortex (50) (17) (18) (50) 

Accessory adrenal cortical nodule 1 (2%) 

Degeneration 1 (2%) 

Hyperplasia 1 (6%) 2 (4%) 

Capsule, hyperplasia, adenomatous 1 (2%) 

Adrenal medulla (50) (17) (18) (48) 

Hyperplasia 3 (6%) 1 (6%) 3 (6%) 

Islets, pancreatic (50) (17) (18) (50) 

Hyperplasia 7 (14%) 1 (6%) 2 (11%) 4 (8%) 

Pituitary gland (50) (17) (18) (49) 

Cyst 1 (2%) 

Pars distalis, hyperplasia 25 (50%) 6 (35%) 11 (61%) 26 (53%) 

Thyroid gland (50) (48) (50) (50) 

Inflammation 1 (2%) 

Follicle, cyst 1 (2%) 

Follicular cell, hyperplasia 26 (52%) 19 (40%) 19 (38%) 25 (50%) 

General Body System

None

Genital System

Ovary (50) (17) (18) (49) 

Cyst 10 (20%) 2 (11%) 13 (27%) 

Hemorrhage 1 (2%) 

Inflammation, chronic active 1 (2%) 

Metaplasia 1 (2%) 

Thrombosis 1 (6%) 

Oviduct (1) 

Inflammation, chronic 1 (100%) 

Uterus (50) (17) (18) (50) 

Hemorrhage 1 (6%) 

Hyperplasia, cystic 21 (42%) 2 (12%) 4 (22%) 22 (44%) 

Pigmentation, hemosiderin 1 (2%) 

Lymphatic, angiectasis 1 (2%) 

Lymphatic, cyst 1 (2%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

198 Sodium Xylenesulfonate, NTP TR 464 

TABLE D5 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Hematopoietic System

Bone marrow (50) (17) (18) (50) 

Hyperplasia 12 (24%) 1 (6%) 4 (22%) 11 (22%) 

Myelofibrosis 1 (2%) 4 (8%) 

Lymph node (7) (3) (6) (6) 

Mediastinal, hyperplasia, lymphoid 1 (14%) 

Mediastinal, infiltration cellular, 

histiocyte 1 (17%) 

Lymph node, mesenteric (48) (17) (17) (50) 

Angiectasis 1 (2%) 2 (4%) 

Hematopoietic cell proliferation 2 (4%) 

Hemorrhage 1 (6%) 

Hyperplasia, lymphoid 1 (2%) 

Inflammation, suppurative 1 (2%) 

Spleen (50) (17) (18) (50) 

Angiectasis 1 (2%) 

Hematopoietic cell proliferation 19 (38%) 8 (47%) 12 (67%) 11 (22%) 

Thymus (38) (16) (16) (44) 

Atrophy 5 (13%) 7 (44%) 4 (25%) 4 (9%) 

Cyst 1 (6%) 

Hyperplasia, lymphoid 2 (5%) 1 (6%) 

Integumentary System

Mammary gland (50) (17) (16) (50) 

Hyperplasia 5 (10%) 1 (6%) 3 (19%) 6 (12%) 

Skin (50) (49) (50) (50) 

Epidermis, skin, site of application, exudate 1 (2%) 4 (8%) 4 (8%) 1 (2%) 

Epidermis, skin, site of application, 

hyperplasia 4 (8%) 1 (2%) 4 (8%) 4 (8%) 

Epidermis, skin, site of application, ulcer 1 (2%) 1 (2%) 1 (2%) 

Skin, site of application, inflammation, 

chronic 4 (8%) 1 (2%) 1 (2%) 1 (2%) 

Skin, site of application, parakeratosis 1 (2%) 

Subcutaneous tissue, edema 1 (2%) 

Subcutaneous tissue, skin, site of

application, inflammation, chronic active 1 (2%) 2 (4%) 

Subcutaneous tissue, skin, site of

application, inflammation, suppurative 1 (2%) 

Musculoskeletal System

None

Nervous System

Brain (50) (17) (18) (50) 

Inflammation 1 (6%) 

Neuron, necrosis 2 (4%) 1 (6%) 4 (22%) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 199 

TABLE D5 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate (continued) 

Vehicle Control 182 mg/kg 364 mg/kg 727 mg/kg

Respiratory System

Lung (50) (18) (18) (50) 

Hemorrhage 1 (2%) 1 (2%) 

Hyperplasia, lymphoid 1 (2%) 

Inflammation 1 (2%) 

Pigmentation, hemosiderin 1 (2%) 

Thrombosis 1 (6%) 

Alveolar epithelium, hyperplasia 2 (4%) 2 (4%) 

Bronchiole, hyperplasia 1 (2%) 1 (2%) 

Nose (50) (17) (18) (50) 

Inflammation, suppurative 1 (2%) 

Special Senses System

Eye (1) (2) 

Degeneration 1 (50%) 

Cornea, inflammation 1 (100%) 

Harderian gland (5) (2) (1) (3) 

Hyperplasia 1 (20%) 

Urinary System

Kidney (50) (17) (18) (50) 

Hydronephrosis 1 (2%) 

Infarct 1 (6%) 

Inflammation, chronic active 1 (2%) 

Mineralization 1 (2%) 

Nephropathy 22 (44%) 1 (6%) 6 (33%) 17 (34%) 

Pigmentation 2 (4%) 

Artery, inflammation, chronic active 1 (6%) 
200 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

201 

APPENDIX E
 

GENETIC TOXICOLOGY
 

SALMONELLA MUTAGENICITY TEST PROTOCOL ...................................... 202 
 #

MOUSE LYMPHOMA MUTAGENICITY TEST PROTOCOL ................................. 202 
 #

CHINESE HAMSTER OVARY CELL CYTOGENETICS PROTOCOLS ............................ 203 
 #

RESULTS .............................................................. 204 
 #

TABLE E1 Mutagenicity of Sodium Xylenesulfonate in Salmonella typhimurium .............. 205 
 #

TABLE E2 Induction of Trifluorothymidine Resistance in L5178Y Mouse Lymphoma Cells
 

by Sodium Xylenesulfonate ......................................... 206 
 #

TABLE E3 Induction of Sister Chromatid Exchanges in Chinese Hamster Ovary Cells
 

by Sodium Xylenesulfonate ......................................... 211 
 #

TABLE E4 Induction of Chromosomal Aberrations in Chinese Hamster Ovary Cells
 

by Sodium Xylenesulfonate ......................................... 213 
 #
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

202 Sodium Xylenesulfonate, NTP TR 464 

GENETIC TOXICOLOGY

SALMONELLA MUTAGENICITY TEST PROTOCOL

Testing was performed as reported by Zeiger et al. (1987). Sodium xylenesulfonate was sent to the

laboratory as a coded aliquot from Radian Corporation (Austin, TX). It was incubated with the Salmonella

typhimurium tester strains (TA98, TA100, TA1535, or TA1537) 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 37E 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 37E C. 

Each trial consisted of triplicate plates of concurrent positive and negative controls and five doses of sodium

xylenesulfonate. The high dose was limited by experimental design to 10,000 μL/plate. All trials were

repeated. 

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 which 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. 

MOUSE LYMPHOMA MUTAGENICITY TEST PROTOCOL

The experimental protocol is presented in detail by Myhr et al. (1985). Sodium xylenesulfonate was

supplied as a coded aliquot by Radian Corporation. The high dose of sodium xylenesulfonate was

determined by toxicity. L5178Y mouse lymphoma cells were maintained at 37E C as suspension cultures in

Fischer’s medium supplemented with l-glutamine, sodium pyruvate, pluronic F68, antibiotics, and heatinactivated horse serum; normal cycling time was approximately 10 hours. To reduce the number of

spontaneously occurring trifluorothymidine-resistant cells, subcultures were exposed to medium containing

THMG (thymidine, hypoxanthine, methotrexate, and glycine) for 1 day, to medium containing THG

(thymidine, hypoxanthine, and glycine) for 1 day, and to normal medium for 3 to 5 days. For cloning, the

horse serum content was increased and Noble agar was added. 

All treatment levels within an experiment, including concurrent positive and solvent controls, were

replicated. Treated cultures contained 6 × 106 cells in 10 mL medium. This volume included the S9 

fraction in those experiments performed with metabolic activation. Incubation with sodium xylenesulfonate

continued for 4 hours, at which time the medium plus sodium xylenesulfonate was removed and the cells

were resuspended in fresh medium and incubated for an additional 2 days to express the mutant phenotype. 

Cell density was monitored so that log phase growth was maintained. After the 48-hour expression period, 

cells were plated in medium and soft agar supplemented with trifluorothymidine (TFT) for selection of TFTresistant (TK!/!

) cells; cells were plated in nonselective medium and soft agar to determine cloning

efficiency. Plates were incubated at 37E C in 5% CO for 2 10 to 12 days. The test was initially performed

without S9. If a clearly positive response was not obtained, the test was repeated using freshly prepared S9 

from the livers of Aroclor 1254-induced male Fischer 344/N rats. 

Minimum criteria for accepting an experiment as valid and a detailed description of the statistical analysis

and data evaluation are presented in Caspary et al. (1988). All data were evaluated statistically for trend

and peak responses. Both responses had to be significant (P#0.05) for sodium xylenesulfonate to be
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 203 

considered positive, i.e., capable of inducing TFT resistance. A single significant response led to a

“questionable” conclusion, and the absence of both a trend and peak response resulted in a “negative” call. 

CHINESE HAMSTER OVARY CELL CYTOGENETICS PROTOCOLS

Testing was performed as reported by Galloway et al. (1987). Sodium xylenesulfonate 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 sodium

xylenesulfonate; in the absence of toxicity, 5 mg/mL was selected as the high dose. A single flask per dose

was used, and tests yielding equivocal or positive results were repeated. 

Sister Chromatid Exchange Test: In the SCE test without S9, CHO cells were incubated for 25.5 hours

with sodium xylenesulfonate in McCoy’s supplemented 5A medium. Bromodeoxyuridine (BrdU) was added

2 hours after culture initiation. After 25.5 hours, the medium containing sodium xylenesulfonate 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 sodium xylenesulfonate, serum-free medium, 

and S9 for 2 hours. The medium was then removed and replaced with medium containing serum and BrdU

and no sodium xylenesulfonate and incubation proceeded for an additional 25.5 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, incubation time was lengthened to ensure a sufficient number of

scorable (second-division metaphase) cells. 

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 sodium xylenesulfonate for 18 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 sodium xylenesulfonate and S9 for 2 hours, after which the treatment medium was

removed and the cells were incubated for 10 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; cell cycle delay was anticipated, 

and 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 at each dose level. 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). 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

204 Sodium Xylenesulfonate, NTP TR 464 

Chromosomal aberration data are presented as percentage 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. 

RESULTS

Sodium xylenesulfonate (100 to 10,000 μg/plate) was not mutagenic in S. typhimiurium strain TA98, 

TA100, TA1535, or TA1537, with and without induced S9 (Zeiger et al., 1987; Table E1). Results

obtained with sodium xylenesulfonate in a mammalian gene mutation assay with cultured L5178Y mouse

lymphoma cells in the presence of S9 (Table E2) were concluded to be equivocal because the significant

increase in mutant colonies noted in the first trial with S9 was not convincingly repeated in the second trial. 

Without S9, no significant increase in mutations was noted. Sodium xylenesulfonate induced dose-related

increases in SCEs in cultured CHO cells at concentrations that produced cell cycle delay (2,513 to

5,000 μg/mL) in the absence of S9; with S9, no increases in SCEs were noted (Table E3). Finally, no

induction of Abs was observed in cultured CHO cells treated with sodium xylenesulfonate (2,513 to

5,000 μg/mL) with or without S9 (Table E4). 
 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 205 

TABLE E1 

Mutagenicity of Sodium Xylenesulfonate in Salmonella typhimuriuma

Revertants/plateb

Strain Dose -S9 +10% hamster S9 +10% rat S9 

(μg/plate) Trial 1 Trial 2 Trial 1 Trial 2 Trial 1 Trial 2 

TA100 0 95 ± 3.9 78 ± 11.2 144 ± 6.2 88 ± 4.5 129 ± 7.5 135 ± 5.8 

100 92 ± 3.6 72 ± 2.7 101 ± 2.5 76 ± 6.7 147 ± 13.7 111 ± 8.1 

333 103 ± 1.5 73 ± 6.1 120 ± 6.0 85 ± 7.5 154 ± 3.5 114 ± 19.7 

1,000 93 ± 1.7 69 ± 3.7 117 ± 1.7 112 ± 14.7 152 ± 2.8 122 ± 5.2 

3,333 90 ± 5.0 69 ± 2.3 119 ± 3.2 100 ± 9.2 112 ± 2.0 103 ± 9.9 

10,000 98 ± 5.0 63 ± 8.5 125 ± 5.6 95 ± 3.2 110 ± 4.7 85 ± 4.5 

Trial summary Negative Negative Negative Negative Negative Negative c Positive control 620 ± 83.1 330 ± 31.2 2,509 ± 107.6 1,708 ± 271.3 1,091 ± 111.2 717 ± 86.1 

TA1535 0 5 ± 1.5 5 ± 0.6 4 ± 1.2 5 ± 0.3 4 ± 0.3 4 ± 2.0 

100 6 ± 0.3 4 ± 0.9 11 ± 0.3 2 ± 1.3 4 ± 2.0 2 ± 1.0 

333 2 ± 0.6 1 ± 0.7 3 ± 1.2 2 ± 1.0 3 ± 0.7 3 ± 0.6 

1,000 2 ± 0.3 3 ± 1.2 7 ± 1.5 5 ± 1.9 6 ± 0.9 4 ± 1.7 

3,333 3 ± 0.3 2 ± 0.9 4 ± 1.5 3 ± 1.5 4 ± 0.6 2 ± 1.0 

10,000 1 ± 0.3 1 ± 0.9 6 ± 0.9 4 ± 2.0 6 ± 0.9 3 ± 1.9 

Trial summary Negative Negative Equivocal Negative Negative Negative

Positive control 108 ± 7.1 134 ± 7.5 93 ± 26.7 48 ± 4.9 115 ± 2.7 56 ± 2.9 

TA1537 0 6 ± 2.4 4 ± 0.7 5 ± 0.6 10 ± 1.5 3 ± 0.5 12 ± 3.0 

100 3 ± 0.9 2 ± 0.3 10 ± 0.0 12 ± 3.2 5 ± 0.7 15 ± 2.7 

333 5 ± 0.6 3 ± 1.2 11 ± 1.8 11 ± 4.6 14 ± 3.0 17 ± 5.9 

1,000 4 ± 0.3 3 ± 0.9 5 ± 0.3 8 ± 0.7 10 ± 1.3 18 ± 1.9 

3,333 2 ± 0.3 2 ± 1.0 9 ± 0.3 13 ± 2.0 9 ± 1.5 18 ± 2.6 

10,000 4 ± 1.9 2 ± 0.9 7 ± 0.3 9 ± 2.0 7 ± 1.5 11 ± 1.3 

Trial summary Negative Negative Negative Negative Equivocal Negative

Positive control 918 ± 78.8 757± 113.8 63 ± 1.3 675 ± 15.3 79 ± 7.9 121 ± 4.1 

TA98 0 15 ± 0.9 12 ± 2.0 23 ± 1.7 19 ± 2.1 20 ± 3.1 26 ± 2.3 

100 15 ± 2.3 17 ± 6.6 24 ± 0.9 20 ± 1.7 24 ± 2.5 22 ± 5.2 

333 19 ± 0.3 16 ± 1.0 18 ± 4.1 19 ± 2.0 20 ± 2.8 23 ± 2.2 

1,000 14 ± 0.9 14 ± 1.7 16 ± 0.7 17 ± 3.8 23 ± 1.5 30 ± 9.5 

3,333 21 ± 3.2 14 ± 1.0 23 ± 0.7 18 ± 1.2 22 ± 3.2 21 ± 3.9 

10,000 23 ± 4.2 13 ± 2.4 26 ± 2.3 17 ± 2.9 26 ± 2.1 24 ± 5.2 

Trial summary Negative Negative Negative Negative Negative Negative

Positive control 301 ± 5.3 269 ± 21.5 2,369 ± 46.4 1,591 ± 75.9 617 ± 103.7 362 ± 25.0 

a Study performed at Case Western Reserve University. The detailed protocol and these data are presented in Zeiger et al. (1987). b	 Revertants are presented as mean ± standard error from three plates. 

The positive controls in the absence of metabolic activation were sodium azide (TA100 and TA1535), 9-aminoacridine (TA1537), and

4-nitro-o-phenylenediamine (TA98). The positive control for metabolic activation with all strains was 2-aminoanthracene. 

c
 

 

206 Sodium Xylenesulfonate, NTP TR 464 

TABLE E2 

Induction of Trifluorothymidine Resistance in L5178Y Mouse Lymphoma Cells

by Sodium Xylenesulfonatea

Compound Concentration

(μg/mL) 

Cloning

Efficiency

(%) 

Relative

Total Growth

(%) 

Mutant

Count

Mutant

Fractionb

Average

Mutant

Fractionc

-S9 

Trial 1 

Dimethylsulfoxide 103 

96 

102 

98 

79 

51 

26 

18 22 

Methyl methanesulfonate 5 79 

98 

60 

47 

60 

36 

381 

403 

446 

160 

137 

246 181* 

Sodium xylenesulfonate 125 93 

86 

66 

91 

67 

63 

64 

65 

60 

23 

25 

30 26 

250 85 

79 

94 

69 

57 

77 

51 

43 

67 

20 

18 

24 21 

500 87 

105 

99 

74 

76 

71 

71 

51 

62 

27 

16 

21 21 

1,000 100 

110 

76 

101 

48 

64 

16 

19 18 

2,000 83 

116 

109 

75 

94 

88 

63 

75 

56 

25 

22 

17 21 

2,500 77 

120 

43 

78 

50 

74 

22 

21 21 
 

 

Sodium Xylenesulfonate, NTP TR 464 207 

TABLE E2 

Induction of Trifluorothymidine Resistance in L5178Y Mouse Lymphoma Cells

by Sodium Xylenesulfonate (continued) 

Compound Concentration

(μg/mL) 

Cloning

Efficiency

(%) 

Relative

Total Growth

(%) 

Mutant

Count

Mutant

Fraction

Average

Mutant

Fraction

-S9 

Trial 2 

Dimethylsulfoxide 70 

79 

67 

80 

83 

105 

91 

121 

47 

62 

47 

58 

23 

26 

23 

24 24 

Methyl methanesulfonate 5 46 

57 

53 

55 

35 

32 

354 

397 

444 

257 

234 

280 257* 

Sodium xylenesulfonate 250 89 

76 

62 

90 

82 

78 

50 

61 

45 

19 

27 

24 23 

500 75 

 77 

72 

74 

80 

75 

69 

47 

56 

31

20 

26 26 

750 80 

91 

77 

73 

75 

73 

66 

46 

65 

28 

17 

28 24 

1,000 80 

93 

75 

80 

90 

78 

42 

51 

62 

18 

18 

28 21 

2,000 86 

 80 

74 

81 

82 

68 

59 

62 

71 

23

26 

32 27 

2,500 79 

 64 

68 

59 

54 

61 

45 

59 

77 

19

31 

38 29 
 

 

 

 

208 Sodium Xylenesulfonate, NTP TR 464 

TABLE E2 

Induction of Trifluorothymidine Resistance in L5178Y Mouse Lymphoma Cells

by Sodium Xylenesulfonate (continued) 

Compound Concentration

(μg/mL) 

Cloning

Efficiency

(%) 

Relative

Total Growth

(%) 

Mutant

Count

Mutant

Fraction

Average

Mutant

Fraction

-S9 

Trial 3 

Supplemented Fischer’s Medium 79 

87 

 80 

89 

114 

100 

93 

93 

72 

72 

88 

67 

31 

27

37 

25 30 

Methyl methanesulfonate 5 39 

30 

34 

25 

5 

13 

435 

352 

267 

377 

391 

260 343* 

Sodium xylenesulfonate 500 72 

57 

64 

22 

69 

81 

32 

48 40 

1,000 77 

108 

 81 

63 

62 

16 

84 

105 

91 

36 

32

38 35 

2,000 57 

79 

Lethal

66 

45 

106 

79 

62 

33 47 

3,000 103 

 84 

92 

41 

27 

74 

80 

64 

79 

26

25 

29 27 

4,000 82 

78 

65 

61 

89 

106 

36 

45 41 

5,000 76 

65 

78 

54 

9 

13 

81 

93 

96 

36 

48 

41 41 
 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 209 

TABLE E2 

Induction of Trifluorothymidine Resistance in L5178Y Mouse Lymphoma Cells

by Sodium Xylenesulfonate (continued) 

Compound Concentration

(μg/mL) 

Cloning

Efficiency

(%) 

Relative

Total Growth

(%) 

Mutant

Count

Mutant

Fraction

Average

Mutant

Fraction

+S9 

Trial 1 

Supplemented Fischer’s Medium 85 

80 

98 

79 

104 

86 

101 

109 

76 

80 

94 

94 

30 

33 

32 

40 34 

Methylcholanthrene 2.5 46 

46 

41 

16 

8 

15 

595 

747 

614 

436 

545 

499 493* 

Sodium xylenesulfonate 250 99 

78 

110 

97 

92 

97 

93 

52 

115 

31 

22 

35 29 

500 85 

103 

 78 

94 

99 

96 

68 

82 

87 

27 

27

37 30 

1,000 69 

 61 

60 

110 

87 

81 

72 

55 

73 

35

30 

41 35 

2,000 76 

109 

108 

52 

81 

63 

85 

107 

87 

37 

33 

27 32 

3,000 83 

90 

 78 

18 

24 

26 

107 

121 

91 

43 

45

39 42 

4,000 102 

75 

Lethal

8 

10 

186 

168 

61 

75 68* 

5,000 Lethal

Lethal

Lethal
 

 

 

 

 

 

 

 

 

210 Sodium Xylenesulfonate, NTP TR 464 

TABLE E2 

Induction of Trifluorothymidine Resistance in L5178Y Mouse Lymphoma Cells

by Sodium Xylenesulfonate (continued) 

Compound Concentration

(μg/mL) 

Cloning

Efficiency

(%) 

Relative

Total Growth

(%) 

Mutant

Count

Mutant

Fraction

Average

Mutant

Fraction

+S9 

Trial 2 

Supplemented Fischer’s Medium 102 

102 

98 

101 

104 

69 

90 

138 

77 

89 

61 

70 

25 

29 

21 

23 25 

Methylcholanthrene 2.5 79 

92 

103 

15 

37 

58 

458 

387 

435 

194 

141 

141 158* 

Sodium xylenesulfonate 250 95 

102 

 82 

130 

133 

60 

40 

88 

53 

14 

29

22 21 

500 107 

98 

114 

153 

100 

140 

55 

106 

70 

17 

36 

20 25 

1,000 114 

104 

106 

95 

120 

111 

96 

82 

78 

28 

26 

25 26 

2,000 95 

109 

104 

51 

69 

72 

107 

106 

86 

38 

32 

28 33 

3,000 101 

95 

Lethal

40 

53 

134 

104 

44 

37 40* 

4,000 Lethal

Lethal

* Significant positive response (P#0.05) a Study performed at Litton Bionetics, Inc. The detailed protocol is presented in Myhr et al. (1985). b Mutant 6 fraction (MF) (frequency) is a ratio of the mutant count to the cloning efficiency, divided by 3 (to arrive at MF/10 cells treated); c Mean from three replicated plates of approximately 106 cells each
 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 211 

TABLE E3 

Induction of Sister Chromatid Exchanges in Chinese Hamster Ovary Cells by Sodium Xylenesulfonatea

Compound

Dose

(μg/mL) 

Total

Cells

Scored

No. of

ChromosomesNo. of

SCEs

SCEs/ 

ChromosomeSCEs/ 

Cell

Relative

Hrs Change of SCEs/ 

in BrdU Chromosome b

(%) 

-S9 

Trial 1 

Summary: Weakly Positive

Negative

50 1,029 392 0.38 7.8 25.5 

Mitomycin-C

0.001 

0.010 

50 

5 

1,036 

105 

556 

192 

0.53 

1.82 

11.1 

38.4 

25.5 

25.5 

40.88 

380.00 

Sodium xylenesulfonate

500 

1,667 

5,000 

50 

50 

50 

1,041 

1,012 

1,023 

436 

432 

528 

0.41 

0.42 

0.51 

8.7 

8.6 

10.6 

25.5 

25.5 

30.7c

9.94 

12.05 

35.48* 

Trial 2 

Summary: Positive

P<0.001d

Negative

25 512 172 0.33 6.9 25.5 

Mitomycin-C

0.001 

0.010 

25 

5 

506 

103 

295 

195 

0.58 

1.89 

11.8 

39.0 

25.5 

25.5 

73.55 

463.57 

Sodium xylenesulfonate

2,513 

3,750 

5,000 

25 

25 

25 

517 

513 

521 

225 

241 

265 

0.43 

0.46 

0.50 

9.0 

9.6 

10.6 

32.5c

32.5c

32.5c

29.55* 

39.84* 

51.41* 

P<0.001 
 

 

 

 

 

 

 

 

 

 

 

212 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE E3 

Induction of Sister Chromatid Exchanges in Chinese Hamster Ovary Cells by Sodium Xylenesulfonate

(continued) 

Compound

Dose

(μg/mL) 

Total

Cells

Scored

No. of

ChromosomesNo. of

SCEs

SCEs/ 

ChromosomeSCEs/ 

Cell

Hrs

in BrdU

Relative

Change of SCEs/ 

Chromosome

(%) 

+S9 

Summary: Negative

Negative

50 1,034 392 0.37 7.8 25.5 

Cyclophosphamide

0.4 

2.0 

50 

5 

1,039 

103 

631 

169 

0.60 

1.64 

12.6 

33.8 

25.5 

25.5 

60.19 

332.80 

Sodium xylenesulfonate

 500 

1,667 

5,000 

50 

50 

50 

1,039 

1,036 

1,032 

413 

391 

396 

0.39 

0.37 

0.38 

P ̆0.532 

8.3 

7.8 

7.9 

25.5 

25.5 

25.5 

4.85 

!0.45 

1.22 

*	#Positive response ($20% increase over solvent control) a Study performed at Litton Bionetics, Inc. A detailed description of the protocol is presented in Galloway et al. (1987). 

SCE ̆sister chromatid exchange; BrdU ̆bromodeoxyuridine b	 SCEs/chromosome in treated cells versus SCEs/chromosome in solvent control cells

Because sodium xylenesulfonate induced a delay in the cell division cycle, harvest time was extended to maximize the proportion of second

division cells available for analysis. d	 Significance of SCEs/chromosome tested by the linear regression trend test versus log of the dose
 

 

 

 

 

 

213 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE E4 

Induction of Chromosomal Aberrations in Chinese Hamster Ovary Cells by Sodium Xylenesulfonatea

-S9 +S9 

Dose

(μg/mL) 

Total

Cells

Scored

No. of

Abs

Abs/ 

Cell

Cells with

Abs (%) 

Dose

(μg/mL) 

Total

Cells

Scored

No. of

Abs

Abs/ 

Cell

Cells with

Abs (%) 

Harvest time: 20.0 hoursb

Summary: Negative

Harvest time: 12.0 hours

Summary: Negative

Negative

200 3 0.02 1.0 

Negative

200 7 0.04 3.5 

Mitomycin-C

0.05 

0.08 

200 

25 

56 

23 

0.28 

0.92 

20.5 

48.0 

Cyclophosphamide

7.5 

37.5 

200 

25 

40 

21 

0.20 

0.84 

16.0 

48.0 

Sodium xylenesulfonate

2,513 200 

3,750 200 

5,000 200 

7 

8 

6 

0.04 

0.04 

0.03 

3.0 

3.5 

3.0 

P ̆0.094c

Sodium xylenesulfonate

2,513 200 

3,750 200 

5,000 200 

9 

8 

8 

0.05 

0.04 

0.04 

4.5 

3.0 

3.5 

P ̆0.612 

a Study performed at Litton Bionetics, Inc. The detailed protocol and these data are presented in Galloway et al. (1987). Abs ̆aberrations. b	 Because of significant chemical-induced cell cycle delay, incubation time prior to the addition of colcemid was lengthened to provide

sufficient metaphase cells at harvest. 

Significance of percent cells with aberrations tested by the linear regression trend test versus log of the dose
214 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

215 

APPENDIX F
 

ORGAN WEIGHTS AND
 

ORGAN-WEIGHT-TO-BODY-WEIGHT RATIOS
 

TABLE F1 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats

in the 17-Day Dermal Study of Sodium Xylenesulfonate ...................... 216 

TABLE F2 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 217 

TABLE F3 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice

in the 17-Day Dermal Study of Sodium Xylenesulfonate ...................... 218 

TABLE F4 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 219 
 

 

 

 

 

 

 

216 Sodium Xylenesulfonate, NTP TR 464 

TABLE F1 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 17-Day Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

n 5 5 5 5 5 5 

Male

Necropsy body wt 181 ± 7 188 ± 6 188 ± 8 174 ± 7 188 ± 12 186 ± 8 

Heart

Absolute 0.690 ± 0.033 0.714 ± 0.036 0.696 ± 0.023 0.662 ± 0.027 0.716 ± 0.037 0.694 ± 0.018 

Relative 3.81 ± 0.13 3.79 ± 0.12 3.74 ± 0.11 3.82 ± 0.10 3.82 ± 0.09 3.75 ± 0.10 

R. Kidney

Absolute 0.972 ± 0.043 1.008 ± 0.031 1.014 ± 0.028 0.962 ± 0.037 1.048 ± 0.062 1.040 ± 0.029 

Relative 5.36 ± 0.08 5.36 ± 0.13 5.45 ± 0.12 5.56 ± 0.18 5.58 ± 0.07 5.63 ± 0.21 

Liver

Absolute 9.488 ± 0.455 9.616 ± 0.274 9.670 ± 0.504 9.274 ± 0.521 10.758 ± 0.700 11.102 ± 0.400 

Relative 52.25 ± 0.87 51.08 ± 0.95 51.72 ± 0.97 53.33 ± 0.87 57.21 ± 0.66** 59.87 ± 1.36** 

Lung

Absolute 0.954 ± 0.015 0.988 ± 0.032 0.954 ± 0.045 1.034 ± 0.039 0.936 ± 0.059 1.120 ± 0.132 

Relative 5.28 ± 0.18 5.25 ± 0.11 5.11 ± 0.16 6.00 ± 0.33 4.99 ± 0.13 6.07 ± 0.75 

R. Testis

Absolute 1.072 ± 0.046 1.004 ± 0.058 1.074 ± 0.066 1.077 ± 0.059 1.032 ± 0.081 1.009 ± 0.041 

Relative 5.91 ± 0.13 5.33 ± 0.24 5.77 ± 0.34 6.20 ± 0.19 5.48 ± 0.19 5.47 ± 0.32 

Thymus

Absolute 0.465 ± 0.013 0.472 ± 0.007 0.439 ± 0.020 0.484 ± 0.018 0.473 ± 0.032 0.462 ± 0.016 

Relative 2.59 ± 0.15 2.51 ± 0.09 2.35 ± 0.07 2.80 ± 0.13 2.52 ± 0.08 2.50 ± 0.08 

Female

Necropsy body wt 136 ± 5 137 ± 5 137 ± 5 133 ± 4 139 ± 5 134 ± 5 

Heart

Absolute 0.550 ± 0.028 0.540 ± 0.016 0.590 ± 0.041 0.550 ± 0.023 0.576 ± 0.011 0.580 ± 0.036 

Relative 4.06 ± 0.17 3.94 ± 0.15 4.31 ± 0.23 4.15 ± 0.13 4.17 ± 0.21 4.34 ± 0.23 

R. Kidney

Absolute 0.748 ± 0.038 0.796 ± 0.035 0.764 ± 0.026 0.744 ± 0.021 0.800 ± 0.014 0.838 ± 0.045 

Relative 5.51 ± 0.15 5.79 ± 0.15 5.58 ± 0.08 5.62 ± 0.09 5.77 ± 0.19 6.26 ± 0.22** 

Liver

Absolute 6.088 ± 0.380 6.290 ± 0.143 6.266 ± 0.154 6.162 ± 0.125 7.160 ± 0.430 6.954 ± 0.309 

Relative 44.74 ± 1.22 45.86 ± 0.89 45.87 ± 0.96 46.58 ± 0.79 51.37 ± 1.90** 52.00 ± 1.40** 

Lung

Absolute 0.856 ± 0.045 0.868 ± 0.037 0.860 ± 0.027 0.806 ± 0.047 0.926 ± 0.045 0.852 ± 0.074 

Relative 6.32 ± 0.28 6.34 ± 0.31 6.29 ± 0.10 6.07 ± 0.22 6.73 ± 0.57 6.42 ± 0.67 

Thymus

Absolute 0.408 ± 0.018 0.393 ± 0.018 0.395 ± 0.008 0.396 ± 0.025 0.353 ± 0.027 0.340 ± 0.025 

Relative 3.02 ± 0.17 2.86 ± 0.07 2.89 ± 0.06 2.98 ± 0.11 2.54 ± 0.21 2.54 ± 0.14 

** Significantly different (P#0.01) from the control group by Williams’ or Dunnett’s test 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). 
 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 217 

TABLE F2 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 14-Week Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

n 10 10 10 10 10 10 

Male

Necropsy body wt 360 ± 8 355 ± 4 354 ± 5 334 ± 6* 342 ± 8 350 ± 6 

Heart

Absolute 1.111 ± 0.025 1.095 ± 0.016 1.042 ± 0.015 1.013 ± 0.024** 1.041 ± 0.025 1.070 ± 0.018 

Relative 3.09 ± 0.05 3.09 ± 0.04 2.95 ± 0.04 3.04 ± 0.06 3.05 ± 0.06 3.06 ± 0.07 

R. Kidney

Absolute 1.429 ± 0.057 1.470 ± 0.020 1.435 ± 0.041 1.362 ± 0.042 1.386 ± 0.035 1.459 ± 0.031 

Relative 3.96 ± 0.09 4.15 ± 0.05 4.06 ± 0.09 4.07 ± 0.07 4.06 ± 0.06 4.17 ± 0.08 

Liver

Absolute 18.010 ± 1.033 16.720 ± 0.303 16.530 ± 0.480 15.426 ± 0.596** 14.856 ± 0.546** 15.904 ± 0.353** 

Relative 49.75 ± 1.94 47.22 ± 1.03 46.70 ± 0.84 46.06 ± 1.02* 43.39 ± 0.75** 45.47 ± 0.74** 

Lung

Absolute 1.466 ± 0.063 1.354 ± 0.017 1.390 ± 0.040 1.297 ± 0.021* 1.310 ± 0.031* 1.365 ± 0.041 

Relative 4.06 ± 0.13 3.82 ± 0.07 3.94 ± 0.12 3.89 ± 0.08 3.84 ± 0.07 3.90 ± 0.10 

R. Testis

Absolute 1.483 ± 0.015 1.488 ± 0.021 1.512 ± 0.015 1.440 ± 0.020 1.486 ± 0.031 1.469 ± 0.021 

Relative 4.13 ± 0.06 4.20 ± 0.07 4.28 ± 0.05 4.32 ± 0.07 4.36 ± 0.09 4.20 ± 0.06 

Thymus

Absolute 0.321 ± 0.027 0.287 ± 0.016 0.304 ± 0.012 0.322 ± 0.021 0.312 ± 0.019 0.317 ± 0.022 

Relative 0.89 ± 0.07 0.81 ± 0.05 0.86 ± 0.04 0.96 ± 0.05 0.91 ± 0.04 0.90 ± 0.05 

Female

Necropsy body wt 196 ± 4 192 ± 3 190 ± 2 192 ± 2 197 ± 4 196 ± 4 

Heart

Absolute 0.724 ± 0.020 0.716 ± 0.016 0.698 ± 0.010 0.728 ± 0.017 0.732 ± 0.016 0.705 ± 0.017 

Relative 3.69 ± 0.05 3.74 ± 0.11 3.68 ± 0.06 3.80 ± 0.07 3.73 ± 0.08 3.60 ± 0.10 

R. Kidney

Absolute 0.874 ± 0.032 0.852 ± 0.021 0.851 ± 0.016 0.840 ± 0.015 0.882 ± 0.028 0.890 ± 0.015 

Relative 4.45 ± 0.11 4.44 ± 0.08 4.49 ± 0.08 4.39 ± 0.07 4.48 ± 0.08 4.55 ± 0.10 

Liver

Absolute 8.007 ± 0.292 7.769 ± 0.222 7.757 ± 0.235 7.877 ± 0.230 7.624 ± 0.160 8.096 ± 0.253 

Relative 40.88 ± 1.30 40.51 ± 1.03 40.90 ± 1.20 41.08 ± 0.92 38.79 ± 0.40 41.24 ± 0.98 

Lung

Absolute 0.997 ± 0.036 1.061 ± 0.029 0.958 ± 0.015 1.037 ± 0.044 0.987 ± 0.018 0.994 ± 0.022 

Relative 5.09 ± 0.14 5.54 ± 0.17 5.06 ± 0.10 5.41 ± 0.22 5.04 ± 0.13 5.07 ± 0.09 

Thymus

Absolute 0.244 ± 0.016 0.250 ± 0.017 0.231 ± 0.014 0.229 ± 0.011 0.257 ± 0.015 0.237 ± 0.012 

Relative 1.24 ± 0.07 1.30 ± 0.09 1.22 ± 0.07 1.20 ± 0.06 1.31 ± 0.07 1.21 ± 0.07 

* 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). 
 

 

 

 

 

 

 

218 Sodium Xylenesulfonate, NTP TR 464 

TABLE F3 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice in the 17-Day Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

n 5 5 5 5 5 5 

Male

Necropsy body wt 25.4 ± 0.8 24.8 ± 1.8 26.1 ± 0.7 25.0 ± 0.3 25.6 ± 0.4 26.4 ± 0.3 

Heart

Absolute 0.140 ± 0.014 0.126 ± 0.010 0.132 ± 0.002 0.132 ± 0.013 0.128 ± 0.004 0.144 ± 0.007 

Relative 5.49 ± 0.48 5.10 ± 0.27 5.06 ± 0.09 5.28 ± 0.50 5.00 ± 0.13 5.45 ± 0.22 

R. Kidney

Absolute 0.268 ± 0.019 0.258 ± 0.025 0.282 ± 0.006 0.264 ± 0.015 0.294 ± 0.011 0.292 ± 0.012 

Relative 10.50 ± 0.48 10.32 ± 0.35 10.83 ± 0.32 10.56 ± 0.52 11.48 ± 0.39 11.05 ± 0.40 

Liver

Absolute 1.200 ± 0.058 1.216 ± 0.136 1.462 ± 0.068* 1.430 ± 0.045* 1.406 ± 0.038 1.746 ± 0.071** 

Relative 47.12 ± 1.11 48.33 ± 2.78 55.93 ± 1.49** 57.29 ± 1.64** 54.93 ± 1.41** 66.10 ± 2.36** 

Lung

Absolute 0.178 ± 0.012 0.166 ± 0.010 0.172 ± 0.002 0.164 ± 0.020 0.166 ± 0.009 0.172 ± 0.007 

Relative 6.99 ± 0.39 6.78 ± 0.48 6.61 ± 0.20 6.56 ± 0.79 6.49 ± 0.33 6.51 ± 0.26 

R. Testis

Absolute 0.105 ± 0.003 0.103 ± 0.003 0.100 ± 0.002 0.098 ± 0.004 0.101 ± 0.003 0.104 ± 0.003 

Relative 4.14 ± 0.09 4.24 ± 0.33 3.82 ± 0.07 3.91 ± 0.13 3.94 ± 0.09 3.92 ± 0.09 

Thymus

Absolute 0.042 ± 0.003 0.047 ± 0.010 0.044 ± 0.006 0.046 ± 0.005 0.049 ± 0.003 0.053 ± 0.005 

Relative 1.66 ± 0.06 1.81 ± 0.32 1.69 ± 0.23 1.83 ± 0.20 1.92 ± 0.12 2.01 ± 0.18 

Female

Necropsy body wt 20.9 ± 0.4 20.2 ± 0.3 20.8 ± 0.5 20.8 ± 0.5 21.0 ± 0.3 21.3 ± 0.2 

Heart

Absolute 0.108 ± 0.004 0.114 ± 0.007 0.116 ± 0.007 0.122 ± 0.009 0.112 ± 0.004 0.138 ± 0.012* 

Relative 5.17 ± 0.22 5.63 ± 0.33 5.58 ± 0.37 5.85 ± 0.29 5.34 ± 0.19 6.46 ± 0.53 

R. Kidney

Absolute 0.182 ± 0.006 0.172 ± 0.007 0.204 ± 0.004 0.196 ± 0.011 0.196 ± 0.010 0.210 ± 0.012 

Relative 8.70 ± 0.21 8.51 ± 0.31 9.81 ± 0.32 9.42 ± 0.33 9.37 ± 0.57 9.84 ± 0.55 

Liver

Absolute 1.132 ± 0.046 1.098 ± 0.041 1.212 ± 0.025 1.234 ± 0.060 1.250 ± 0.051 1.418 ± 0.054** 

Relative 54.07 ± 1.70 54.24 ± 1.33 58.24 ± 1.33 59.32 ± 1.78 59.52 ± 1.87* 66.43 ± 2.33** 

Lung

Absolute 0.152 ± 0.004 0.158 ± 0.012 0.174 ± 0.015 0.164 ± 0.014 0.158 ± 0.011 0.204 ± 0.016* 

Relative 7.27 ± 0.19 7.81 ± 0.56 8.39 ± 0.79 7.86 ± 0.52 7.54 ± 0.55 9.58 ± 0.82 

Thymus

Absolute 0.085 ± 0.004 0.072 ± 0.008 0.068 ± 0.005 0.080 ± 0.002 0.085 ± 0.003 0.080 ± 0.004 

Relative 4.08 ± 0.21 3.56 ± 0.37 3.26 ± 0.27 3.86 ± 0.08 4.05 ± 0.16 3.76 ± 0.21 

* 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). 
 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP 464 219 

TABLE F4 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice in the 14-Week Dermal Study

of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

Male

n 10 10 10 10 10 10 

Necropsy body wt 36.0 ± 0.6 32.9 ± 0.8* 35.6 ± 0.7 36.7 ± 0.6 35.5 ± 0.7 37.4 ± 0.7 

Heart

Absolute 0.173 ± 0.006 0.161 ± 0.004 0.172 ± 0.005 0.171 ± 0.005 0.166 ± 0.007 0.170 ± 0.004 

Relative 4.81 ± 0.14 4.92 ± 0.15 4.86 ± 0.20 4.66 ± 0.11 4.70 ± 0.23 4.56 ± 0.10 

R. Kidney

Absolute 0.329 ± 0.010 0.354 ± 0.008 0.363 ± 0.008* 0.375 ± 0.011** 0.352 ± 0.008 0.370 ± 0.009** 

Relative 9.16 ± 0.26 10.82 ± 0.33** 10.22 ± 0.20* 10.21 ± 0.23* 9.96 ± 0.36 9.93 ± 0.26 

Liver

Absolute 1.698 ± 0.046 1.575 ± 0.045 1.682 ± 0.034 1.741 ± 0.054 1.644 ± 0.021 1.748 ± 0.051 

Relative 47.24 ± 1.12 48.10 ± 1.63 47.33 ± 0.53 47.37 ± 1.18 46.38 ± 0.72 46.76 ± 0.75 

Lung

Absolute 0.191 ± 0.011 0.178 ± 0.006 0.192 ± 0.009 0.194 ± 0.008 0.190 ± 0.008 0.184 ± 0.006 

Relative 5.32 ± 0.30 5.45 ± 0.24 5.41 ± 0.25 5.29 ± 0.23 5.37 ± 0.24 4.93 ± 0.14 

R. Testis

Absolute 0.126 ± 0.003 0.124 ± 0.002 0.122 ± 0.002 0.124 ± 0.002 0.120 ± 0.002 0.124 ± 0.002 

Relative 3.49 ± 0.08 3.79 ± 0.06 3.43 ± 0.08 3.39 ± 0.07 3.38 ± 0.09 3.34 ± 0.06 

Thymus

Absolute 0.046 ± 0.002 0.034 ± 0.002** 0.044 ± 0.002 0.044 ± 0.002 0.042 ± 0.002 0.046 ± 0.002 

Relative 1.27 ± 0.06 1.03 ± 0.07* 1.23 ± 0.07 1.19 ± 0.06 1.18 ± 0.06 1.22 ± 0.03 

Female

n 10 9 9 10 9 10 

Necropsy body wt 30.6 ± 0.8 30.2 ± 0.6 30.8 ± 0.7 32.5 ± 0.7 30.7 ± 0.6 30.1 ± 0.8 

Heart

Absolute 0.137 ± 0.002 0.137 ± 0.004 0.132 ± 0.002 0.139 ± 0.003 0.143 ± 0.006 0.138 ± 0.004 

Relative 4.49 ± 0.10 4.54 ± 0.14 4.30 ± 0.08 4.29 ± 0.10 4.67 ± 0.15 4.60 ± 0.12 

R. Kidney

Absolute 0.237 ± 0.004 0.239 ± 0.004 0.236 ± 0.006 0.238 ± 0.005 0.228 ± 0.004 0.234 ± 0.004 

Relative 7.77 ± 0.16 7.94 ± 0.16 7.65 ± 0.16 7.36 ± 0.23 7.44 ± 0.17 7.81 ± 0.17 

Liver

Absolute 1.474 ± 0.038 1.484 ± 0.028 1.418 ± 0.036 1.384 ± 0.022 1.462 ± 0.056 1.442 ± 0.035 

Relative 48.19 ± 0.74 49.31 ± 0.92 46.02 ± 0.81 42.70 ± 0.80** 47.59 ± 1.28 48.11 ± 1.14 

Lung

Absolute 0.184 ± 0.006 0.184 ± 0.005 0.170 ± 0.007 0.176 ± 0.006 0.183 ± 0.003 0.173 ± 0.004 

Relative 6.03 ± 0.20 6.13 ± 0.17 5.52 ± 0.23 5.43 ± 0.20 5.99 ± 0.13 5.79 ± 0.19 

Thymus

Absolute 0.057 ± 0.002 0.049 ± 0.002 0.053 ± 0.002 0.052 ± 0.003 0.053 ± 0.004 0.051 ± 0.001 

Relative 1.86 ± 0.04 1.63 ± 0.05 1.73 ± 0.07 1.61 ± 0.09 1.74 ± 0.11 1.71 ± 0.05 

* 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). 
220 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

221 

APPENDIX G

HEMATOLOGY

AND CLINICAL CHEMISTRY RESULTS

TABLE G1 Hematology Data for Male Rats

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 222 

TABLE G2 Hematology Data for Female Rats

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 223 

TABLE G3 Clinical Chemistry Data for Male Rats

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 224 

TABLE G4 Clinical Chemistry Data for Female Rats

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 225 
 

 

 

 

 

 

222 Sodium Xylenesulfonate, NTP TR 464 

TABLE G1 

Hematology Data for Male Rats in the 14-Week Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

n

Day 5 7 10 9 10 8 6 

Day 21 10 10 9 10 10 9 

Week 14 10 10 10 10 10 10 

Hematocrit (%) 

Day 5 44.7 ± 0.6 42.7 ± 0.5 43.1 ± 0.6 43.4 ± 0.3 43.8 ± 0.7 42.6 ± 0.7 

Day 21 45.6 ± 1.1 46.4 ± 0.3 44.2 ± 2.3 48.0 ± 0.5 45.9 ± 0.4 45.8 ± 0.5 

Week 14 47.7 ± 0.5 47.5 ± 0.5 47.0 ± 0.5 46.8 ± 0.3 47.4 ± 0.3 46.7 ± 0.6 

Hemoglobin (g/dL) 

Day 5 14.9 ± 0.2 14.3 ± 0.2 14.4 ± 0.3 14.6 ± 0.1 14.7 ± 0.2 14.2 ± 0.1 

Day 21 15.0 ± 0.3 15.2 ± 0.1 14.6 ± 0.8 15.9 ± 0.2 15.2 ± 0.1 15.1 ± 0.2 

Week 14 15.7 ± 0.2 15.6 ± 0.2 15.5 ± 0.1 15.6 ± 0.1 15.5 ± 0.1 15.5 ± 0.1 

Erythrocytes (10 /μL) 6 

Day 5 7.51 ± 0.08 7.14 ± 0.06 7.38 ± 0.14 7.37 ± 0.06 7.34 ± 0.19 6.98 ± 0.16* 

Day 21 8.09 ± 0.16 8.24 ± 0.07 7.96 ± 0.39 8.55 ± 0.09 8.19 ± 0.14 8.05 ± 0.09 

Week 14 9.63 ± 0.08 9.67 ± 0.09 9.41 ± 0.09 9.45 ± 0.08 9.49 ± 0.07 9.42 ± 0.13 

Reticulocytes (10 /μL) 6 

Day 5 0.03 ± 0.01b 0.03 ± 0.01 0.04 ± 0.01 0.03 ± 0.01 0.04 ± 0.01 0.02 ± 0.00 

Day 21 0.07 ± 0.02 0.06 ± 0.01 0.08 ± 0.02 0.09 ± 0.01 0.06 ± 0.02 0.08 ± 0.01 

Week 14 0.02 ± 0.00 0.01 ± 0.00 0.02 ± 0.00 0.01 ± 0.00 0.01 ± 0.00 0.01 ± 0.00 

Mean cell volume (fL) 

Day 5 59.6 ± 0.7 59.9 ± 0.6 58.3 ± 0.4 58.9 ± 0.5 59.9 ± 0.7 61.0 ± 0.6 

Day 21 56.5 ± 0.5 56.5 ± 0.4 55.3 ± 0.6 56.1 ± 0.3 56.1 ± 0.6 57.1 ± 0.4 

Week 14 49.5 ± 0.2 49.1 ± 0.2 49.9 ± 0.2 49.4 ± 0.2 50.1 ± 0.2 49.6 ± 0.2 

Mean cell hemoglobin (pg) 

Day 5 19.9 ± 0.3 20.0 ± 0.3 19.5 ± 0.1 19.8 ± 0.2 20.0 ± 0.4 20.4 ± 0.4 

Day 21 18.5 ± 0.2 18.5 ± 0.1 18.2 ± 0.2 18.5 ± 0.2 18.6 ± 0.2 18.8 ± 0.2 

Week 14 16.3 ± 0.1 16.1 ± 0.1 16.5 ± 0.1 16.5 ± 0.1 16.3 ± 0.1 16.5 ± 0.1 

Mean cell hemoglobin concentration (g/dL) 

Day 5 33.4 ± 0.2 33.4 ± 0.2 33.3 ± 0.3 33.6 ± 0.2 33.5 ± 0.3 33.4 ± 0.3 

Day 21 32.8 ± 0.2 32.7 ± 0.2 32.9 ± 0.3 33.0 ± 0.2 33.1 ± 0.2 33.0 ± 0.3 

Week 14 32.9 ± 0.2 32.7 ± 0.1 33.0 ± 0.2 33.4 ± 0.2 32.7 ± 0.2 33.2 ± 0.3 

Platelets (10 /μL) 3 

Day 5 987.7 ± 33.4 980.4 ± 33.3 959.8 ± 37.7 984.8 ± 63.7 914.3 ± 48.4 878.3 ± 15.9* 

Day 21 746.2 ± 55.3 743.9 ± 55.0 843.8 ± 121 817.7 ± 21.8 708.1 ± 37.5 835.0 ± 30.7 

Week 14 702.9 ± 17.1 729.9 ± 21.2 699.1 ± 18.0 699.9 ± 18.1 715.2 ± 8.6 678.3 ± 12.2 

Leukocytes (10 /μL) 3 

Day 5 4.93 ± 0.81 3.91 ± 0.31 3.67 ± 0.44 4.08 ± 0.20 3.70 ± 0.51 3.58 ± 0.45 

Day 21 5.72 ± 0.34 4.78 ± 0.47 6.47 ± 1.26 6.92 ± 0.30 5.61 ± 0.56 6.57 ± 0.55 

Week 14 8.49 ± 0.38 8.78 ± 0.29 8.59 ± 0.30 9.86 ± 0.47 8.96 ± 0.17 9.02 ± 0.34 

Segmented neutrophils (10 /μL) 3 

Day 5 0.64 ± 0.18 0.54 ± 0.05 0.48 ± 0.08 0.42 ± 0.06 0.55 ± 0.10 0.47 ± 0.10 

Day 21 0.59 ± 0.10 0.61 ± 0.15 0.74 ± 0.23 0.47 ± 0.04 0.69 ± 0.09 0.63 ± 0.11 

Week 14 1.33 ± 0.09 1.12 ± 0.11 1.00 ± 0.05 1.08 ± 0.11 1.11 ± 0.10 1.28 ± 0.13 

Lymphocytes (10 /μL) 3 

Day 5 4.21 ± 0.67 3.36 ± 0.27 3.13 ± 0.40 3.65 ± 0.19 3.14 ± 0.43 3.12 ± 0.40 

Day 21 5.06 ± 0.24 4.16 ± 0.44 5.67 ± 1.02 6.40 ± 0.31 4.88 ± 0.48 5.87 ± 0.51 

Week 14 6.64 ± 0.30 6.99 ± 0.26 7.08 ± 0.30 8.04 ± 0.49 7.17 ± 0.10 7.09 ± 0.30 

Monocytes (10 /μL) 3 

Week 14 0.46 ± 0.08 0.56 ± 0.08 0.42 ± 0.07 0.58 ± 0.06 0.55 ± 0.06 0.56 ± 0.08 

Eosinophils (10 /μL) 3 

Day 5 0.04 ± 0.03 0.03 ± 0.02 0.02 ± 0.02 0.02 ± 0.01 0.00 ± 0.00 0.02 ± 0.02 

Day 21 0.06 ± 0.04 0.03 ± 0.02 0.06 ± 0.03 0.06 ± 0.02 0.05 ± 0.02 0.07 ± 0.02 

Week 14 0.09 ± 0.03 0.12 ± 0.04 0.11 ± 0.04 0.15 ± 0.03 0.15 ± 0.05 0.09 ± 0.04 

* Significantly different (P#0.05) from the control group by Dunn’s or Shirley’s test a Mean ± standard error. Statistical tests were performed on unrounded data. b n ̆6 
 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 223 

TABLE G2 

Hematology Data for Female Rats in the 14-Week Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

n

Day 5 6 10 7 10 8 8 

Day 21 10 10 10 10 10 10 

Week 14 10 10 10 10 10 10 

Hematocrit (%) 

Day 5 45.5 ± 0.8 43.6 ± 0.7 43.9 ± 0.5 44.8 ± 0.4 44.3 ± 0.9 43.0 ± 0.7 

Day 21 46.3 ± 0.7 46.3 ± 0.4 46.6 ± 0.7 46.6 ± 0.5 44.7 ± 0.8 46.2 ± 0.5 

Week 14 46.3 ± 0.4 46.7 ± 0.4 47.6 ± 0.4 46.8 ± 0.4 46.9 ± 0.4 46.4 ± 0.3 

Hemoglobin (g/dL) 

Day 5 14.8 ± 0.2 14.5 ± 0.3 14.6 ± 0.2 14.8 ± 0.2 14.5 ± 0.3 14.4 ± 0.3 

Day 21 15.5 ± 0.2 15.5 ± 0.2 15.4 ± 0.2 15.5 ± 0.2 15.0 ± 0.2 15.4 ± 0.2 

Week 14 15.3 ± 0.1 15.4 ± 0.1 15.6 ± 0.1 15.4 ± 0.1 15.4 ± 0.1 15.4 ± 0.2 

Erythrocytes (10 /μL) 6 

Day 5 7.83 ± 0.17 7.38 ± 0.13 7.51 ± 0.11 7.49 ± 0.07 7.50 ± 0.16 7.32 ± 0.11 

Day 21 7.97 ± 0.13 7.96 ± 0.09 8.02 ± 0.13 7.96 ± 0.11 7.78 ± 0.11 8.02 ± 0.10 

Week 14 8.65 ± 0.07 8.71 ± 0.07 8.91 ± 0.06* 8.78 ± 0.07 8.77 ± 0.07 8.67 ± 0.06 

Reticulocytes (10 /μL) 6 

Day 5 0.12 ± 0.01 0.15 ± 0.01 0.12 ± 0.02 0.08 ± 0.01 0.08 ± 0.01 0.12 ± 0.02 

Day 21 0.07 ± 0.01 0.07 ± 0.01 0.10 ± 0.02 0.11 ± 0.02* 0.10 ± 0.01* 0.10 ± 0.01* 

Week 14 0.05 ± 0.02 0.03 ± 0.01 0.02 ± 0.00 0.03 ± 0.01 0.04 ± 0.01 0.04 ± 0.01 

Mean cell volume (fL) 

Day 5 58.2 ± 0.5 59.1 ± 0.6 58.6 ± 0.5 59.9 ± 0.6 59.3 ± 0.5 58.8 ± 0.6 

Day 21 58.1 ± 0.4 58.3 ± 0.4 58.2 ± 0.3 58.5 ± 0.4 57.5 ± 0.5 57.7 ± 0.4 

Week 14 53.6 ± 0.2 53.6 ± 0.2 53.5 ± 0.2 53.3 ± 0.2 53.5 ± 0.2 53.3 ± 0.2 

Mean cell hemoglobin (pg) 

Day 5 19.0 ± 0.2 19.6 ± 0.3 19.5 ± 0.1 19.7 ± 0.2 19.4 ± 0.2 19.7 ± 0.3 

Day 21 19.5 ± 0.1 19.5 ± 0.1 19.2 ± 0.2 19.5 ± 0.1 19.3 ± 0.2 19.2 ± 0.1 

Week 14 17.7 ± 0.1 17.7 ± 0.1 17.5 ± 0.1 17.6 ± 0.1 17.6 ± 0.1 17.8 ± 0.1 

Mean cell hemoglobin concentration (g/dL) 

Day 5 32.6 ± 0.2 33.2 ± 0.3 33.3 ± 0.2 33.0 ± 0.2 32.7 ± 0.2 33.5 ± 0.2* 

Day 21 33.5 ± 0.2 33.5 ± 0.3 33.1 ± 0.2 33.3 ± 0.3 33.6 ± 0.2 33.4 ± 0.3 

Week 14 32.9 ± 0.2 33.0 ± 0.2 32.8 ± 0.2 33.0 ± 0.2 32.9 ± 0.3 33.3 ± 0.1 

Platelets (10 /μL) 3 

Day 5 802.8 ± 28.4 925.2 ± 42.0 846.1 ± 13.8 863.9 ± 20.0 893.9 ± 26.5 877.1 ± 15.7 

Day 21 700.1 ± 30.5 742.3 ± 36.1 748.3 ± 32.8 837.2 ± 12.2** 792.9 ± 29.5* 831.7 ± 17.0** 

Week 14 752.9 ± 31.4 699.2 ± 8.4 713.5 ± 10.5 705.7 ± 16.4 715.5 ± 16.1 708.2 ± 16.6 

Leukocytes (10 /μL) 3 

Day 5 3.77 ± 0.77 4.38 ± 0.49 4.14 ± 0.61 4.16 ± 0.66 4.16 ± 0.57 4.39 ± 0.54 

Day 21 3.94 ± 0.46 4.46 ± 0.48 3.29 ± 0.28 4.37 ± 0.51 4.81 ± 0.52 4.42 ± 0.44 

Week 14 6.16 ± 0.47 5.42 ± 0.45 5.68 ± 0.45 5.41 ± 0.48 6.02 ± 0.43 6.16 ± 0.37 

Segmented neutrophils (10 /μL) 3 

Day 5 0.47 ± 0.09 0.28 ± 0.04 0.49 ± 0.14 0.45 ± 0.12 0.61 ± 0.10 0.54 ± 0.13 

Day 21 0.39 ± 0.07 0.56 ± 0.12 0.32 ± 0.03 0.35 ± 0.06 0.53 ± 0.08 0.58 ± 0.10 

Week 14 0.86 ± 0.21 0.63 ± 0.09 0.74 ± 0.14 0.71 ± 0.10 0.74 ± 0.12 0.82 ± 0.09 

Lymphocytes (10 /μL) 3 

Day 5 3.27 ± 0.69 4.04 ± 0.49 3.64 ± 0.48 3.69 ± 0.57 3.53 ± 0.48 3.85 ± 0.44 

Day 21 3.49 ± 0.40 3.90 ± 0.41 2.96 ± 0.26 4.01 ± 0.46 4.25 ± 0.48 3.80 ± 0.38 

Week 14 5.23 ± 0.34 4.73 ± 0.38 4.90 ± 0.39 4.68 ± 0.41 5.17 ± 0.42 5.25 ± 0.32 

Monocytes (10 /μL) 3 

Week 14 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.00 ± 0.00 0.02 ± 0.02 

Eosinophils (10 /μL) 3 

Day 5 0.07 ± 0.03 0.03 ± 0.02 0.03 ± 0.02 0.02 ± 0.01 0.04 ± 0.02 0.04 ± 0.02 

Day 21 0.04 ± 0.02 0.02 ± 0.01 0.00 ± 0.00 0.00 ± 0.00 0.02 ± 0.01 0.03 ± 0.02 

Week 14 0.07 ± 0.03 0.06 ± 0.03 0.05 ± 0.02 0.04 ± 0.02 0.09 ± 0.03 0.07 ± 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. 
 

 

 

224 Sodium Xylenesulfonate, NTP TR 464 

TABLE G3 

Clinical Chemistry Data for Male Rats in the 14-Week Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

n

Day 5 10 10 10 10 10 9 

Day 21 10 10 10 10 10 9 

Week 14 10 10 10 10 10 10 

Urea nitrogen (mg/dL) 

Day 5 28.0 ± 1.2 23.6 ± 0.6** 26.0 ± 0.8 25.7 ± 0.8 24.6 ± 0.7 24.3 ± 0.9 

Day 21 25.9 ± 1.0 25.9 ± 1.0 25.8 ± 1.4 25.5 ± 0.4 25.6 ± 1.2 24.4 ± 1.0 

Week 14 23.4 ± 0.6 23.9 ± 0.7 23.3 ± 0.6 23.2 ± 0.4 23.0 ± 0.7 23.5 ± 0.7 

Creatinine (mg/dL) 

Day 5 0.41 ± 0.02 0.46 ± 0.02 0.43 ± 0.02 0.44 ± 0.04 0.46 ± 0.02 0.41 ± 0.03 

Day 21 0.62 ± 0.03 0.60 ± 0.02 0.61 ± 0.02 0.61 ± 0.02 0.60 ± 0.02 0.56 ± 0.05 

Week 14 0.50 ± 0.02 0.50 ± 0.02 0.49 ± 0.02 0.50 ± 0.01 0.50 ± 0.01 0.47 ± 0.03 

Total protein (g/dL) 

Day 5 6.0 ± 0.1 6.0 ± 0.1 6.1 ± 0.2 6.1 ± 0.1 6.0 ± 0.1 5.8 ± 0.1 

Day 21 6.3 ± 0.1 6.3 ± 0.2 5.9 ± 0.2 6.6 ± 0.1 6.1 ± 0.1 6.2 ± 0.1 

Week 14 6.9 ± 0.1 7.0 ± 0.1 6.8 ± 0.1 6.8 ± 0.1 7.0 ± 0.1 6.8 ± 0.1 

Albumin (g/dL) 

Day 5 3.6 ± 0.1 3.5 ± 0.1 3.7 ± 0.1 3.6 ± 0.1 3.6 ± 0.1 3.6 ± 0.0 

Day 21 3.6 ± 0.1 3.6 ± 0.1 3.5 ± 0.1 3.7 ± 0.1 3.6 ± 0.1 3.5 ± 0.1 

Week 14 3.9 ± 0.1 4.0 ± 0.1 3.9 ± 0.1 3.9 ± 0.1 3.9 ± 0.1 3.8 ± 0.1 

Alanine aminotransferase (IU/L) 

Day 5 41 ± 1 45 ± 1 42 ± 1 48 ± 2** 47 ± 2** 45 ± 1* 

Day 21 59 ± 7 55 ± 3 52 ± 2 52 ± 3 52 ± 2 47 ± 2 

Week 14 52 ± 2 59 ± 5 52 ± 1 54 ± 2 51 ± 1 55 ± 3 

Alkaline phosphatase (IU/L) 

Day 5 579 ± 26 617 ± 25 569 ± 22 593 ± 13 615 ± 25 595 ± 12 

Day 21 468 ± 19 471 ± 18 459 ± 17 464 ± 8 460 ± 16 457 ± 13 

Week 14 258 ± 9 274 ± 12 270 ± 7 272 ± 9 276 ± 8 267 ± 9 

Creatine kinase (IU/L) 

Day 5 531 ± 38 407 ± 56 428 ± 73 608 ± 177 612 ± 149 373 ± 46 

Day 21 334 ± 61 296 ± 39 296 ± 38 445 ± 117 291 ± 31 272 ± 25 

Week 14 287 ± 30 228 ± 31 276 ± 37 264 ± 58 202 ± 26 231 ± 62 

Sorbitol dehydrogenase (IU/L) 

Day 5 6 ± 0 6 ± 0 5 ± 0 7 ± 1 6 ± 1 6 ± 1 

Day 21 7 ± 1 6 ± 0 6 ± 0 7 ± 1 6 ± 1 6 ± 0 

Week 14 8 ± 0 11 ± 2 9 ± 1 8 ± 0 7 ± 0 9 ± 1 

Bile acids (μmol/L) 

Day 5 19.3 ± 3.0 18.4 ± 2.9 18.3 ± 3.0 28.6 ± 3.1 19.2 ± 5.0 23.1 ± 3.4 

Day 21 18.9 ± 2.7 28.8 ± 6.8 28.4 ± 5.1 25.7 ± 3.6 26.2 ± 3.7 28.6 ± 3.5 

Week 14 30.0 ± 5.3 18.2 ± 2.6 21.9 ± 4.9 21.3 ± 3.1 23.2 ± 4.3 21.4 ± 3.2 

* 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. 
 

 

Sodium Xylenesulfonate, NTP TR 464 225 

TABLE G4 

Clinical Chemistry Data for Female Rats in the 14-Week Dermal Study of Sodium Xylenesulfonatea

Vehicle Control 5 mg/mL 15 mg/mL 44 mg/mL 133 mg/mL 400 mg/mL

n 10 10 10 10 10 10 

Urea nitrogen (mg/dL) 

Day 5 25.3 ± 0.7 25.8 ± 0.8 26.7 ± 0.5 27.0 ± 0.5 26.4 ± 0.5 27.1 ± 1.0 

Day 21 27.3 ± 1.1 26.7 ± 0.8 28.1 ± 0.9 26.1 ± 0.8 27.2 ± 0.6 26.1 ± 0.6 

Week 14 23.7 ± 1.0 23.3 ± 0.8 23.9 ± 1.1 25.0 ± 0.6 25.0 ± 0.8 24.9 ± 0.8 

Creatinine (mg/dL) 

Day 5 0.42 ± 0.03 0.42 ± 0.04 0.47 ± 0.02 0.49 ± 0.03b 0.45 ± 0.02 0.37 ± 0.05 

Day 21 0.59 ± 0.02 0.58 ± 0.02 0.63 ± 0.02 0.58 ± 0.02 0.56 ± 0.03 0.58 ± 0.02 

Week 14 0.44 ± 0.02 0.44 ± 0.03 0.47 ± 0.02 0.48 ± 0.01 0.45 ± 0.01 0.48 ± 0.02 

Total protein (g/dL) 

Day 5 5.7 ± 0.1 5.6 ± 0.1 5.8 ± 0.1 5.7 ± 0.1 5.8 ± 0.1 5.7 ± 0.1 

Day 21 6.0 ± 0.2 6.1 ± 0.1 6.2 ± 0.1 6.1 ± 0.2 5.9 ± 0.1 6.2 ± 0.1 

Week 14 6.4 ± 0.1 6.4 ± 0.1 6.5 ± 0.0 6.5 ± 0.1 6.5 ± 0.1 6.4 ± 0.1 

Albumin (g/dL) 

Day 5 3.6 ± 0.0 3.5 ± 0.1 3.7 ± 0.1 3.6 ± 0.1 3.7 ± 0.1 3.5 ± 0.0 

Day 21 3.6 ± 0.0 3.5 ± 0.1 3.6 ± 0.1 3.5 ± 0.1 3.4 ± 0.1 3.6 ± 0.1 

Week 14 3.7 ± 0.1 3.7 ± 0.1 3.8 ± 0.1 3.8 ± 0.1 3.8 ± 0.1 3.8 ± 0.1 

Alanine aminotransferase (IU/L) 

Day 5 43 ± 2 42 ± 1 42 ± 1 41 ± 2 43 ± 2 41 ± 1 

Day 21 49 ± 2 46 ± 2 45 ± 2 43 ± 2 46 ± 2 45 ± 2 

Week 14 45 ± 2 44 ± 1 47 ± 1 46 ± 1 47 ± 2 50 ± 3 

Alkaline phosphatase (IU/L) 

Day 5 508 ± 12 509 ± 12 536 ± 17 514 ± 12 469 ± 15 500 ± 12 

Day 21 437 ± 13 408 ± 13 423 ± 6 439 ± 8 387 ± 15 422 ± 12 

Week 14 274 ± 8 285 ± 8 285 ± 12 285 ± 10 290 ± 14 294 ± 10 

Creatine kinase (IU/L) 

Day 5 449 ± 57 423 ± 40 441 ± 72 419 ± 38 345 ± 25b 563 ± 83 

Day 21 463 ± 20 407 ± 39 463 ± 44 436 ± 31 548 ± 58 475 ± 47 

Week 14 230 ± 42 223 ± 38 291 ± 74 219 ± 19 281 ± 72 335 ± 110 

Sorbitol dehydrogenase (IU/L) 

Day 5 3 ± 0 4 ± 0 4 ± 0 4 ± 1 4 ± 1 4 ± 0 

Day 21 4 ± 0 4 ± 0 4 ± 0 4 ± 0 3 ± 0 4 ± 0 

Week 14 6 ± 0 6 ± 0 6 ± 0 5 ± 0 6 ± 0 6 ± 0 

Bile acids (μmol/L) 

Day 5 32.0 ± 4.9 28.6 ± 4.8 32.0 ± 8.2 22.0 ± 3.5 31.5 ± 5.5 22.9 ± 3.3 

Day 21 20.0 ± 3.8 20.3 ± 4.7 25.1 ± 4.7 26.8 ± 4.7 17.3 ± 1.4 22.4 ± 3.0 

Week 14 22.3 ± 3.8 20.6 ± 4.5 23.2 ± 4.8 21.6 ± 4.0 15.9 ± 2.9 27.1 ± 3.4 

a Mean ± standard error. Statistical tests were performed on unrounded data. b n ̆9 
226 Sodium Xylenesulfonate, NTP TR 464 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

227 

APPENDIX H
 

CHEMICAL CHARACTERIZATION AND
 

DOSE FORMULATION STUDIES
 

PROCUREMENT AND CHARACTERIZATION OF SODIUM XYLENESULFONATE .................... 228 
 #

PREPARATION AND ANALYSIS OF DOSE FORMULATIONS ................................ 229 
 #

FIGURE H1 Infrared Absorption Spectrum of Sodium Xylenesulfonate .................... 230 
 #

FIGURE H2 Nuclear Magnetic Resonance Spectrum of Sodium Xylenesulfonate .............. 231 
 #

TABLE H1 Preparation and Storage of Dose Formulations in the Dermal Studies
 

of Sodium Xylenesulfonate .......................................... 232 
 #

TABLE H2 Results of Analyses of Dose Formulations Administered to Rats and Mice
 

in the 17-Day Dermal Studies of Sodium Xylenesulfonate ..................... 233 
 #

TABLE H3 Results of Analyses of Dose Formulations Administered to Rats and Mice
 

in the 14-Week Dermal Studies of Sodium Xylenesulfonate .................... 234 
 #

TABLE H4 Results of Analyses of Dose Formulations Administered to Rats and Mice
 

in the 2-Year Dermal Studies of Sodium Xylenesulfonate ..................... 235 
 #

TABLE H5 Results of Referee Analyses of Dose Formulations Administered
 

to Rats and Mice in the 14-Week Dermal Studies of Sodium Xylenesulfonate ........ 238 
 #
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

228 Sodium Xylenesulfonate, NTP TR 464 

CHEMICAL CHARACTERIZATION AND

DOSE FORMULATION

PROCUREMENT AND CHARACTERIZATION

OF SODIUM XYLENESULFONATE

Sodium xylenesulfonate was obtained from Ruetgers Nease Chemical Company (State College, PA) in one

lot (R092085), which was used for the 17-day, 14-week, and 2-year studies. Identity, purity, and stability

studies were conducted by the analytical chemistry laboratory, Midwest Research Institute (Kansas City, 

MO). Reports on analyses performed in support of the sodium xylenesulfonate studies are on file at the

National Institutes of Environmental Health Sciences. 

The chemical, a white powder, was identified as sodium xylenesulfonate by infrared, ultraviolet/visible, and

nuclear magnetic resonance spectroscopy. All spectra were consistent with those expected for the structure; 

infrared and nuclear magnetic resonance spectra were consistent with the literature spectra (Sadtler Standard

Spectra) for sodium xylenesulfonate (Figures H1 and H2). 

The purity was determined by elemental analysis, Karl Fischer water analysis, functional group titration, 

thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC). Functional group

titration for ionic sulfate was performed by dissolving the sodium xylenesulfonate sample in a water:acetone

solution and titrating with 0.01 N barium perchlorate solution. The titration was monitored visually with

Sulfonazo III indicator. TLC was performed on Silica Gel 60 F-254 plates with two solvent systems: 

1) chloroform:methanol (50:50) and 2) acetone:water (90:10). 2-Naphthalenesulfonic acid was used as a

reference standard. Plates were examined under visible and under ultraviolet light (254 nm) and iodine

vapor. Plates were also inspected under visible and ultraviolet light (254 nm and 366 nm) after a spray of

silver nitrate/sodium fluorescein. HPLC was performed with a Waters μBondapak phenyl column at a flow

of 1 mL/min with UV detection at 254 nm. Two isocratic solvent systems were used consisting of

(A) 0.005 M tetrabutylammonium hydroxide in water with the pH adjusted to 7 with phosphoric acid and

(B) 0.005 M tetrabutylammonium hydroxide in methanol with an identical volume of phosphoric acid

added. The solvent ratio used in system 1 was 70:30 (A:B) and, in system 2, 40:60. 

Elemental analyses for carbon and hydrogen were in agreement with the theoretical values for sodium

xylene sulfonate. Calculated on the basis of 5.15% water and 3.82% sodium sulfate, results for sulfur were

slightly higher than theoretical values and results for sodium were slightly lower than theoretical values. 

Karl Fischer water analysis indicated 5.15% ± 0.07% water. Functional group titration indicated

3.82% ± 0.04% sodium sulfate. TLC by system 1 indicated a major spot and a slight trace impurity. TLC

by system 2 indicated a major spot only. HPLC by system A indicated a major peak and five impurities

with areas totalling 39.2% relative to the major peak; system B indicated an additional impurity with a peak

area of 9.6% relative to the major peak. 

Under these chromatographic conditions, sodium 2,4- and 2,5-xylenesulfonate coelute with the major peak. 

Evidence for sodium ethyl benzenesulfonate being present as a component in the material can be seen in the

nuclear magnetic resonance spectrum, (i.e., triplet at 1.14 ppm [CH2

CH3

] and quartet at approximately

2.5 ppm [CH2CH ]) due to the ethyl group of ethyl benzenesulfonate. 3 

Concomitant HPLC analyses of lot R092085 with lot 3835 (not used in these studies) indicated a purity of

approximately 115% for lot R092085 relative to lot 3835. 

Stability studies of lot 3835 were performed by the analytical chemistry laboratory. To ensure stability of

the bulk chemical during the 17-day and 14-week studies, sodium xylenesulfonate was stored in glass bottles
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 229 

with Teflon®-lined caps or double bagged in metal drums at room temperature in the dark. During the

2-year studies the bulk chemical was stored in amber glass bottles at room temperature in the dark. 

PREPARATION AND ANALYSIS OF DOSE FORMULATIONS

The dose formulations for the 17-day studies were prepared twice during the study by stirring the

appropriate quantities of sodium xylenesulfonate to solution in deionized water and were then brought to the

desired concentration by the further addition of deionized water. The dose formulations for the 14-week

studies were prepared every 2 weeks by mixing the appropriate amount of sodium xylenesulfonate with 50% 

ethanol (in deionized water) which was then brought to the desired concentration by the further addition of

50% ethanol. The 5 and 15 mg/mL doses were stirred to solution and the 44, 133, and 400 mg/mL doses

were stirred to suspension. Dose formulations in the 2-year studies of sodium xylenesulfonate were

prepared every 2 to 3 weeks by mixing the appropriate weight of sodium xylenesulfonate in deionized

water. A suspension was formed by shaking, and then the mixture was brought to the desired

concentrations of ethanol and sodium xylenesulfonate by the addition of 95% ethanol. The resulting

suspension was the required dose in 50% ethanol. Dose formulations in the 17-day, 14-week, and 2-year

studies were stored at room temperature in glass bottles with Teflon®-lined lids in the dark for up to 3 weeks

(17-day and 14-week studies) or for 3 to 4 weeks (2-year studies). 

Stability studies of 4 mg/mL dose formulations (in deionized water and in 50% ethanol) were conducted by

the analytical chemistry laboratory. Aliquots (20 mL) of the solutions were analyzed by HPLC with a

Zorbax C column, a mobile phase of cetyltrimethylammonium bromide (10 mg/mL water): methanol 8 

(50:50 for dosed water; 25:75 for ethanol formulations), a flow rate of 1 mL/min, and benzoic acid as an

internal standard; detection was at 254 nm. Stability was confirmed for at least 3 weeks when stored in the

dark at room temperature in sealed glass vials and for 3 hours at room temperature open to air and light. 

Homogeneity studies of the dose formulations used in the 2-year studies and a limited stability study of the

75 mg/mL dose formulation used in the 2-year studies were performed by the study laboratory using the

same HPLC system as in the previous dose stability study. Homogeneity was confirmed; the 75 mg/mL

concentration was stable for 29 days when stored at room temperature, protected from light. 

Periodic analyses of the dose formulations of sodium xylenesulfonate were conducted by the study

laboratory with HPLC. During the 17-day studies, doses were analyzed at the beginning of the studies

(Table H2). During the 14-week studies, doses were analyzed at the beginning, midpoint, and end of the

studies (Table H3). During the 2-year studies, dose formulations were analyzed at the beginning of the

studies and every 7 to 10 weeks thereafter (Table H4). Although the results of a preliminary mixing trial

for the 5 and 400 mg/mL concentrations used in the 17-day studies were within 10% of the target

concentrations, the first set of dose formulations were 12% to 14% greater than the target concentrations

due to a mixing error. These formulations were remixed and were all within 10% of the target

concentrations (Table H2). Animal-room samples for the 17-day studies were also within 10% of the target

concentrations. During the 14-week studies, all of the dose formulations and all of the animal room samples

were within 10% of the target concentrations. In the 2-year studies, all of the dose formulations and all but

two of the animal room samples were within 10% of the target concentrations. For the 14-week studies, 

two referee analyses performed by the analytical chemistry laboratory indicated good agreement with the

results of the study laboratory (Table H5). 
 

230 Sodium Xylenesulfonate, NTP TR 464
 

FIGURE H1 

Infrared Absorption Spectrum of Sodium Xylenesulfonate 
 

Sodium Xylenesulfonate, NTP TR 464
 231 

FIGURE H2 

Nuclear Magnetic Resonance Spectrum of Sodium Xylenesulfonate 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

232 Sodium Xylenesulfonate, NTP TR 464 

TABLE H1 

Preparation and Storage of Dose Formulations in the Dermal Studies

of Sodium Xylenesulfonate

17-Day Studies 14-Week Studies 2-Year Studies

Preparation

The required weight of sodium The required weight of sodium The required weight of sodium

xylenesulfonate was stirred to solution in xylenesulfonate was mixed with 50% xylenesulfonate was mixed with

deionized water. The solution was ethanol (in deionized water). The deionized water by shaking. The

brought to the desired concentrations by formulation was brought to the desired resulting suspension was then brought to

the further addition of deionized water. concentration by the further addition of the desired concentration of ethanol

50% ethanol. The 5 and 15 mg/mL (50%) and sodium xylenesulfonate by

doses were stirred to solution while the the addition of 95% ethanol. 

44, 133, and 400 mg/mL doses were

stirred to suspension. 

Chemical Lot Number

R092085 R092085 R092085 

Maximum Storage Time

20 days 20 days 3 to 4 weeks

Storage Conditions

Stored at room temperature in the dark Same as 17-day studies Same as 17-day studies

Study Laboratory

Southern Research Institute Southern Research Institute Battelle Columbus Laboratories

(Birmingham, AL) (Birmingham, AL) (Columbus, OH) 

Referee Laboratory

None Midwest Research Institute None

(Kansas City, MO) 
 

 

 

 

 

 

 

233 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE H2 

Results of Analyses of Dose Formulations Administered to Rats and Mice in the 17-Day Dermal Studies

of Sodium Xylenesulfonate

Date Prepared Date Analyzed

Target

Concentration

(mg/mL) 

Determined

Concentrationa 

(mg/mL) 

% Difference

from Target

5 June 1987b 8-9 June 1987 5 

400 

5.14 

412 

+3 

+3 

15 July 1987 16 July 1988 5 

15 

44 

133 

400 

5.66 

16.9 

49.1 

152 

452 

+13 

+13 

+12 

+14 

+13 

17 July 1987c 20 July 1987 5 

15 

44 

133 

400 

4.94 

15.0 

44.4 

134 

408 

!1 

0 

+1 

+1 

+2 

17 July 1987 28-29 July 1987d 5 

15 

44 

133 

400 

5.05 

15.3 

45.2 

138 

414 

+1 

+2 

+3 

+4 

+4 

17 July 1987 28-29 July 1987e 5 

15 

44 

133 

400 

5.17 

15.2 

45.0 

136 

412 

+3 

+1 

+2 

+2 

+3 

a Results of duplicate analyses. Dosing volume ̆300 μL for rats and 100 μL for mice. b Preliminary mixing trial; not used for dosing

Results of remix

d Animal room samples for rats e Animal room samples for mice
 

 

 

 

234 Sodium Xylenesulfonate, NTP TR 464 

TABLE H3 

Results of Analyses of Dose Formulations Administered to Rats and Mice in the 14-Week Dermal Studies

of Sodium Xylenesulfonate

Target Determined % Difference

Date Prepared Date Analyzed Concentration Concentrationa from Target

(mg/mL) (mg/mL) 

10 February 1988 11-12 February 1988 5 5.12 +2 

15 15.3 +2 

44 44.6 +1 

133 134 +1 

400 425 +6 

29 February - 2 March 1988b 5 5.09 +2 

15 15.2 +1 

44 44.0 0 

133 132 !1 

400 406 +2 

23 March 1988 23-25 March 1988 5 5.02 0 

15 14.8 !1 

44 43.9 0 

133 135 +2 

400 410 +3 

12-13 April 1988b 5 5.08 +2 

15 14.8 !1 

44 43.9 0 

133 133 0 

400 410 +3 

4 May 1988 6-9 May 1988 5 4.96 !1 

15 15.2 +1 

44 43.9 0 

133 132 !1 

400 406 +2 

26-27 May 1988b 5 5.01 0 

15 15.1 +1 

44 43.8 0 

133 132 !1 

400 398 0 

a

b

Results of duplicate analyses. Dosing volume ̆300 μL for rats and 100 μL for mice. 

Animal room sample
 

 

 

Sodium Xylenesulfonate, NTP TR 464 235 

TABLE H4 

Results of Analyses of Dose Formulations Administered to Rats and Mice in the 2-Year Dermal Studies

of Sodium Xylenesulfonate

Target Determined % Difference a Date Prepared Date Analyzed Concentration Concentration from Target

(mg/mL) (mg/mL) 

Rats

26 November 1990 28 November 1990 75 74.2 !1 

150 146 !3 

300 296 !1 

b 26 November 1990 13 December 1990 75 76.1 +1 

150 150 0 

300 303 +1 

Rats and Mice

6 December 1990 14 December 1990 75 74.8 0 

150 150 0 

300 293 !2 

c 6 December 1990 4 January 1991 75 75.0 0 

150 152 +1 

300 296 !1 

14 February 1991 18 February 1991 75 76.3 +2 

150 155 +3 

300 299 0 

11 April 1991 30 April 1991 75 79.5 +6 

150 154 +3 

300 317 +6 

11 April 1991b 30 April 1991 75 79.1 +5 

150 157 +5 

300 312 +4 

c 11 April 1991 11 April 1991 75 78.5 +5 

150 159 +6 

300 308 +3 

6 June 1991 11 June 1991 75 77.6 +3 

150 155 +3 

300 301 0 

1 August 1991 2 August 1991 75 76.9 +3 

150 155 +3 

300 309 +3 

26 September 1991 27 September 1991 75 74.4 !1 

150 146 !3 

300 298 !1 

26 September 1991b 28 October 1991 75 78.1 +4 

150 155 +3 

300 308 +3 
 

 

 

236 Sodium Xylenesulfonate, NTP TR 464 

TABLE H4 

Results of Analyses of Dose Formulations Administered to Rats and Mice in the 2-Year Dermal Studies

of Sodium Xylenesulfonate (continued) 

Date Prepared Date Analyzed

Target

Concentration

(mg/mL) 

Determined

Concentration

(mg/mL) 

% Difference

from Target

Rats and Mice (continued) 

26 September 1991c 28 October 1991 75 

150 

300 

78.4 

154 

309 

+5 

+3 

+3 

21 November 1991 22 November 1991 75 

150 

300 

77.1 

154 

302 

+3 

+3 

+1 

16 January 1992 17 January 1992 75 

150 

300 

76.5 

152 

298 

+2 

+1 

!1 

12 March 1992 12 March 1992 75 

150 

300 

76.3 

152 

302 

+2 

+1 

+1 

12 March 1992b 9-10 April 1992 75 

150 

300 

78.8 

162 

306 

+5 

+8 

+2 

12 March 1992c 9-10 April 1992 75 

150 

300 

79.6 

156 

306 

+6 

+4 

+2 

7 May 1992 8 May 1992 75 

150 

300 

76.5 

153 

305 

+2 

+2 

+2 

2 July 1992 6 July 1992 75 

150 

300 

77.4 

152 

301 

+3 

+1 

0 

20 August 1992 21 August 1992 75 

150 

300 

75.6 

156 

308 

+1 

+4 

+3 

20 August 1992b 21-22 September 1992 75 

150 

300 

77.3 

157 

312 

+3 

+5 

+4 

20 August 1992c 21-22 September 1992 75 

150 

300 

79.9 

171 

333 

+7 

+14 

+11 
 

 

 

 

 

 

 

237 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE H4 

Results of Analyses of Dose Formulations Administered to Rats and Mice in the 2-Year Dermal Studies

of Sodium Xylenesulfonate (continued) 

Target Determined % Difference

Date Prepared Date Analyzed Concentration Concentration from Target

(mg/mL) (mg/mL) 

Rats and Mice (continued) 

22 October 1992 26-27 October 1992 75 79.7 +6 

150 153 +2 

300 326 +9 

a Results of duplicate analyses. Dosing volume for rats was 85 to 357 μL; 60 mg/kg  ̆ 75 mg/mL; 120 mg/kg  ̆ 150 mg/mL; 

240 mg/kg  ̆ 300 mg/mL; dosing volume for mice was 46 to 128 μL; 182 mg/kg  ̆ 75 mg/mL; 364 mg/kg  ̆ 150 mg/mL; 

727 mg/kg  ̆ 300 mg/mL. b	 Animal room samples for rats

Animal room samples for mice
 

 

 

 

238 Sodium Xylenesulfonate, NTP TR 464 

TABLE H5 

Results of Referee Analyses of Dose Formulations Administered to Rats and Mice

in the 14-Week Dermal Studies of Sodium Xylenesulfonate

Date Prepared

Target Concentration

(mg/mL) 

Determined Concentration (mg/mL) 

Study Referee

Laboratory Laboratory a b

10 February 1988 

23 March 1988 

133 

15.0 

134 

14.8 

132 ± 1 

14.8 ± 0.0 

a Results of duplicate analyses b Results of triplicate analyses (mean ± standard error) 
 

 

 

 

 

 

 

 

 

 

 

 

 

239 

APPENDIX I

DOSES, BODY WEIGHTS, 

AND DOSE CONCENTRATIONS

TABLE I1 

TABLE I2 

TABLE I3 

TABLE I4 

TABLE I5 

TABLE I6 

Doses, Body Weights, and Dose Concentrations for Rats

in the 17-Day Dermal Study of Sodium Xylenesulfonate ...................... 

Doses, Body Weights, and Dose Concentrations for Mice

in the 17-Day Dermal Study of Sodium Xylenesulfonate ...................... 

Doses, Body Weights, and Dose Concentrations for Rats

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 

Doses, Body Weights, and Dose Concentrations for Mice

in the 14-Week Dermal Study of Sodium Xylenesulfonate ..................... 

Doses, Body Weights, and Dose Concentrations for Rats

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 

Doses, Body Weights, and Dose Concentrations for Mice

in the 2-Year Dermal Study of Sodium Xylenesulfonate ...................... 

240 

240 

241 

241 

242 

242 
 

 

 

 

 

 

240 Sodium Xylenesulfonate, NTP TR 464
#

TABLE I1 

Doses, Body Weights, and Dose Concentrations for Rats in the 17-Day Dermal Study

of Sodium Xylenesulfonate

Dose Dose Volume Average Body Weight Dose Concentration

(mg/mL) (μL) (kg) (mg/kg) 

Male

0 300 0.145 0
#

5 300 0.150 10
#

15 300 0.150 30
#

44 300 0.140 94
#

133 300 0.151 264
#

400 300 0.149 808
#

Female

0 300 0.114 0
#

5 300 0.117 13
#

400 300 0.116 1,035
#

15 300 0.118 38
#

44 300 0.113 117
#

133 300 0.120 334
#

TABLE I2 

Doses, Body Weights, and Dose Concentrations for Mice in the 17-Day Dermal Study

of Sodium Xylenesulfonate

Dose Dose Volume Average Body Weight Dose Concentration

(mg/mL) (μL) (kg) (mg/kg) 

Male

0 100 0.025 0
#

5 100 0.025 20
#

400 100 0.025 1,633
#

15 100 0.025 61
#

44 100 0.024 187
#

133 100 0.025 532
#

Female

0 100 0.020 0
#

5 100 0.019 26
#

400 100 0.020 2,051
#

15 100 0.020 77
#

44 100 0.020 220
#

133 100 0.020 682
#
 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 241 

TABLE I3 

Doses, Body Weights, and Dose Concentrations for Rats in the 14-Week Dermal Study

of Sodium Xylenesulfonate

Weeks 1 to 4 Weeks 4 to 8 Weeks 8 to 14 

Dose Dose Volume Avg Wt Dose Concentration Avg Wt Dose Concentration Avg Wt Dose Concentration

(mg/mL) (mL) (kg) (mg/kg) (kg) (mg/kg) (kg) (mg/kg) 

Male

0 300 0.192 0 0.265 0 0.322 0 

5 300 0.189 8 0.266 6 0.323 5 

15 300 0.179 25 0.259 17 0.322 14 

44 300 0.178 74 0.255 52 0.308 43 

133 300 0.180 222 0.255 157 0.316 126 

400 300 0.179 670 0.254 472 0.319 376 

Female

0 300 0.130 0 0.164 0 0.187 0 

5 300 0.123 12 0.158 10 0.181 8 

15 300 0.121 37 0.156 29 0.180 25 

44 300 0.122 108 0.160 83 0.183 72 

133 300 0.128 312 0.163 245 0.188 212 

400 300 0.128 938 0.163 736 0.186 645 

TABLE I4 

Doses, Body Weights, and Dose Concentrations for Mice in the 14-Week Dermal Study

of Sodium Xylenesulfonate

Weeks 1 to 4 Weeks 4 to 8 Weeks 8 to 14 

Dose Dose Volume Avg Wt Dose Concentration Avg Wt Dose Concentration Avg Wt Dose Concentration

(mg/mL) (mL) (kg) (mg/kg) (kg) (mg/kg) (kg) (mg/kg) 

Male

0 100 0.027 0 0.030 0 0.034 0 

5 100 0.027 19 0.029 17 0.033 15 

15 100 0.027 56 0.030 50 0.034 44 

44 100 0.028 157 0.031 142 0.035 126 

133 100 0.027 493 0.030 443 0.034 391 

400 100 0.028 1,429 0.031 1,290 0.035 1,143 

Female

0 100 0.021 0 0.025 0 0.029 0 

5 100 0.021 24 0.025 20 0.028 18 

15 100 0.021 71 0.025 60 0.029 52 

44 100 0.022 200 0.026 169 0.030 147 

133 100 0.022 605 0.025 532 0.028 475 

400 100 0.022 1,818 0.025 1,600 0.028 1,429 
 

 

 

 

 

 

242 Sodium Xylenesulfonate, NTP TR 464 

TABLE I5 

Doses, Body Weights, and Dose Concentrations for Rats in the 2-Year Dermal Study

of Sodium Xylenesulfonate

Dose

(mg/mL) 

Dose Volume

(μL) 

Average Body Weight Range

(kg) 

Dose Concentration

(mg/kg) 

Male

0 

75 

150 

300 

102 ! 357 

101 ! 357 

102 ! 349 

102 ! 345 

0.127 ! 0.439 

0.127 ! 0.438 

0.128 ! 0.437 

0.127 ! 0.432 

0 

60 

120 

240 

Female

0 

75 

150 

300 

86 ! 232 

86 ! 230 

85 ! 232 

86 ! 217 

0.108 ! 0.289 

0.107 ! 0.288 

0.107 ! 0.290 

0.107 ! 0.271 

0 

60 

120 

240 

TABLE I6 

Doses, Body Weights, and Dose Concentrations for Mice in the 2-Year Dermal Study

of Sodium Xylenesulfonate

Dose

(mg/mL) 

Dose Volume

(μL) 

Average Body Weight Range

(kg) 

Dose Concentration

(mg/kg) 

Male

0 

75 

150 

300 

58 ! 116 

57 ! 114 

58 ! 113 

57 ! 116 

0.024 ! 0.047 

0.024 ! 0.047 

0.024 ! 0.047 

0.024 ! 0.048 

0 

182 

364 

727 

Female

0 

75 

150 

300 

46 ! 128 

46 ! 128 

46 ! 122 

46 ! 128 

0.019 ! 0.051 

0.019 ! 0.052 

0.019 ! 0.050 

0.019 ! 0.053 

0 

182 

364 

727 
 

 

 

 

 

243 

APPENDIX J

INGREDIENTS, NUTRIENT COMPOSITION, 

AND CONTAMINANT LEVELS

IN NIH-07 RAT AND MOUSE RATION

TABLE J1 

TABLE J2 

TABLE J3 

TABLE J4 

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 ....................... 

244 

244 

245 

246 
 

 

 

 

 

 

 

 

 

 

 

 

 

244 Sodium Xylenesulfonate, NTP TR 464 

TABLE J1 

Ingredients of NIH-07 Rat and Mouse Rationa

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 J2 

Vitamins and Minerals in NIH-07 Rat and Mouse Rationa

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
 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 245 

TABLE J3 

Nutrient Composition of NIH-07 Rat and Mouse Ration

Mean ± Standard

Nutrient Deviation Range Number of Samples

Protein (% by weight) 23.45 ± 0.52 22.2 – 24.3 23 

Crude fat (% by weight) 5.34 ± 0.19 5.00 – 5.90 23 

Crude fiber (% by weight) 3.37 ± 0.34 2.60 – 4.30 23 

Ash (% by weight) 6.46 ± 0.18 6.12 – 6.81 23 

Amino Acids (% of total diet) 

Arginine 1.280 ± 0.083 1.110 – 1.390 11 

Cystine 0.308 ± 0.071 1.181 – 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) 6,780 ± 1,363 5,730 – 11,450 23 

Vitamin D (IU/kg) 4,450 ± 1,382 3,000 – 6,300 4 

"-Tocopherol (ppm) 36.12 ± 9.15 22.5 – 48.9 10 

Thiamine (ppm) 17.48 ± 2.06 14.0 – 22.0 23 

Riboflavin (ppm) 7.83 ± 0.923 6.10 – 9.00 11 

Niacin (ppm) 98.64 ± 25.51 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 B (ppb) 40.5 ± 19.1 10.6 – 65.0 11 12 

Choline (ppm) 2,991 ± 382 2,300 – 3,430 10 

Minerals

Calcium (%) 1.17 ± 0.09 1.00 – 1.49 23 

Phosphorus (%) 0.92 ± 0.05 0.760 – 1.00 23 

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 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

246 Sodium Xylenesulfonate, NTP TR 464 

TABLE J4 

Contaminant Levels in NIH-07 Rat and Mouse Rationa

Mean ± Standard

Deviationb Range Number of Samples

Contaminants

Arsenic (ppm) 0.48 ± 0.17 0.10 – 0.70 23 

Cadmium (ppm) 0.14 ± 0.07 0.04 – 0.20 23 

Lead (ppm) 0.36 ± 0.25 0.10 – 1.00 23 

Mercury (ppm) 0.02 ± 0.00 0.02 – 0.03 23 

Selenium (ppm) 0.32 ± 0.11 0.05 – 0.04 23 

Aflatoxins (ppb) <5.0 23 c Nitrate nitrogen (ppm) 8.07 ± 4.02 2.90 – 17.0 23 c Nitrite nitrogen (ppm) 0.15 ± 0.07 0.10 – 0.30 23 

BHA (ppm)

d 1.48 ± 0.95 1.00 – 5.00 23 

BHT (ppm)

d 1.35 ± 0.88 1.00 – 5.00 23 

Aerobic plate count (CFU/g) 102,443 ± 168,151 4,100 – 710,000 23 

Coliform (MPN/g) 3.1 ± 0.3 3.0 – 4.0 23 

Escherichia coli (MPN/g) <3.0 23 

Salmonella (MPN/g) Negative 23 e Total nitrosoamines (ppb) 7.53 ± 1.84 4.70 – 11.40 23 e N-Nitrosodimethylamine (ppb) 5.58 ± 1.22 2.90 – 8.20 23 e N-Nitrosopyrrolidine (ppb) 1.96 ± 1.04 1.00 – 4.30 23 

Pesticides (ppm) 

"-BHC <0.01 23 

$-BHC <0.02 23 

(-BHC <0.01 23 

*-BHC <0.01 23 

Heptachlor <0.01 23 

Aldrin <0.01 23 

Heptachlor epoxide <0.01 23 

DDE <0.01 23 

DDD <0.01 23 

DDT <0.01 23 

HCB <0.01 23 

Mirex <0.01 23 

Methoxychlor <0.05 23 

Dieldrin <0.01 23 

Endrin <0.01 23 

Telodrin <0.01 23 

Chlordane <0.05 23 

Toxaphene <0.10 23 

Estimated PCBs <0.20 23 

Ronnel <0.01 23 

Ethion <0.02 23 

Trithion <0.05 23 

Diazinon <0.10 23 

Methyl parathion <0.02 23 

Ethyl parathion <0.02 23 

Malathion 0.23 ± 0.24 0.05 – 0.97 23 

Endosulfan I <0.01 23 

Endosulfan II <0.01 23 

Endosulfan sulfate <0.03 23 

a CFU  ̆ colony-forming units, MPN  ̆ most probable number, BHC  ̆ hexachlorocyclohexane or benezene hexachloride

b For values less than the limit of detection, the detection limit is given as the mean. 

Sources of contamination: alfalfa, grains, and fish meal

d Sources of contamination: soy oil and fish meal e All values were corrected for percent recovery. 

c
 

 

 

247 

APPENDIX K

SENTINEL ANIMAL PROGRAM

METHODS

TABLE K1 

.............................................................. 

Murine Virus Antibody Determinations for Rats and Mice

in the 14-Week and 2-Year Studies of Sodium Xylenesulfonate ................. 

248 

250 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

248 Sodium Xylenesulfonate, NTP TR 464 

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 rodents is monitored via serology on sera from extra

(sentinel) animals in the study rooms. These animals and the study animals are all 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 and mice during the 14-week 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 the blood was collected during the studies are also listed. 

Method and Test Time of Analysis

RATS

14-Week Study

ELISA

PVM (pneumonia virus of mice) 

RCV/SDA

(rat coronavirus/sialodacryoadenitis virus) 

Sendai

14 weeks

14 weeks

14 weeks

Hemagglutination Inhibition

H-1 (Toolan’s H-1 virus) 

KRV (Kilham rat virus) 

14 weeks

14 weeks

2-Year Study

ELISA

Mycoplasma arthritidis 24 months

Mycoplasma pulmonis 24 months

PVM Quarantine, 6, 12, 18, and 24 months

RCV/SDA Quarantine, 6, 12, 18, and 24 months

Sendai Quarantine, 6, 12, 18, and 24 months

Hemagglutination Inhibition

Quarantine, 6, 12, 18, and 24 months

KRV Quarantine, 6, 12, 18, and 24 months

H-1 
 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 249 

MICE

14-Week Study

ELISA

Ectromelia virus

GDVII (mouse encephalomyelitis virus) 

LCM (lymphocytic choriomeningitis virus) 

MVM (minute virus of mice) 

Mouse adenoma virus

MHV (mouse hepatitis virus) 

PVM

Sendai

Immunofluorescence Assay

EDIM (epizootic diarrhea of infant mice) 

Reovirus 3 

Hemagglutination Inhibition

K (papovavirus) 

Polyoma virus

2-Year Study

ELISA

Ectromelia virus

EDIM

GDVII

LCM

Mouse adenoma virus-FL

MHV

M. arthritidis

M. pulmonis

PVM
 

Reovirus 3
#

Sendai
 

Immunofluorescence Assay

EDIM

LCM

Mouse adenoma virus-FL

MHV

Reovirus 3 

Hemagglutination Inhibition

K

MVM

Polyoma virus

Results of serology tests are presented in Table K1. 

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

14 weeks

Quarantine, 6, 12, 18, and 24 months

Quarantine, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months

24 months

24 months

Quarantine, 6, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months

6, 12, and 18 months

18 months

18 months

12 and 18 months

12 and 18 months

Quarantine, 6, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months

Quarantine, 6, 12, 18, and 24 months
 

 

 

 

 

 

 

 

250 Sodium Xylenesulfonate, NTP TR 464 

TABLE K1 

Murine Virus Antibody Determinations for Rats and Mice in the 14-Week and 2-Year Dermal Studies

of Sodium Xylenesulfonate

Interval Incidence of Antibody

in Sentinel Animals

Positive Serologic

Reaction for

14-Week Studies

Rats

Study termination 0/10 None positive

Mice

Study termination 0/10 None positive

2-Year Studies

Rats

Quarantine

6 Months

12 Months

18 Months

24 Months

0/10 

0/10 

0/9 

0/7 

2/10 

None positive

None positive

None positive

None positive

M. arthritidisa

Mice

Quarantine

6 Months

12 Months

18 Months

24 Months

0/9 

0/10 

1/10 

0/9 

2/10 

None positive

None positive

Reovirus 3 

None positive

M. arthritidisa

a Further evaluation of samples positive for M. arthritidis by immunoblot and Western blot procedures indicated that the positive titers may

have been due to cross reaction with antibiotics of nonpathogenic Mycoplasma or other agents. Only sporadic samples were positive, and

there were no clinical findings or histopathologic changes of M. athritidis infection in rats or mice with positive titers. Accordingly, sporadic

M. arthritidis-positive titers were considered to be false positives. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

251 

APPENDIX L
 

IMPACT OF HELICOBACTER HEPATICUS
 

INFECTION IN B6C3F1 MICE

FROM 12 NTP 2-YEAR CARCINOGENESIS STUDIES
 

1 James R. Hailey , Joseph K. Haseman1 1 1
# , John R. Bucher , Ann E. Radovsky , 2 2 1 David E. Malarkey , Richard T. Miller , Abraham Nyska , and Robert R. Maronpot1
#

1

National Institute of Environmental Health Sciences
 

Research Triangle Park, North Carolina
 

2

Department of Microbiology, Pathology, and Parasitology
 

College of Veterinary Medicine
 

North Carolina State University
 

Raleigh, North Carolina
 

ABSTRACT .............................................................. 252 
 #

INTRODUCTION ........................................................... 252 
 #

MATERIALS AND METHODS ................................................... 253 
 #

RESULTS AND DISCUSSION .................................................... 255 
 #

REFERENCES ............................................................ 262 
 #

TABLE L1 Incidence of Helicobacter hepaticus-Associated Hepatitis in Control B6C3F1 Mice

from Nine NTP 2-Year Studies ...................................... 266 
 #

TABLE L2 Identification of Helicobacter hepaticus with PCR-RFLP-Based Assays
 

in Control B6C3F Mice from 32 NTP 2-Year Studies 1 

and Three NTP 13-Week Studies ..................................... 266 
 #

TABLE L3 Comparison of Neoplasm Incidences in Control B6C3F1
#Mice

from Helicobacter hepaticus-Affected and Unaffected NTP 2-Year Studies ......... 267 
 #

TABLE L4 Liver Neoplasm Incidences and Body Weights of Control B6C3F1
#Mice

in Relation to Study Start Dates of Helicobacter hepaticus-Affected

and Unaffected NTP 2-Year Studies ................................... 268 
 #

TABLE L5 Association of Liver Neoplasm Incidence
 

and Severity of Helicobacter hepaticus-Associated Hepatitis

in Control B6C3F Mice from Nine Affected NTP 2-Year Studies 1 . . . . . . . . . . . . . . . 269 
 #

TABLE L6	# H-ras Codon 61 AAA Mutations in Spontaneous Liver Neoplasms

in Control B6C3F1 Mice from Helicobacter hepaticus-Affected

and Unaffected NTP 2-Year Studies ................................... 269 
 #

TABLE L7 Proliferating Cell Nuclear Antigen Labeling Indices in the Liver
 

of Control B6C3F1 Mice ........................................... 
 # 270 

TABLE L8 Summary of Target Sites of Carcinogenicity in B6C3F1
#Mice

from NTP 2-Year Studies with Helicobacter hepaticus-Associated Hepatitis ........ 271 
 #
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

252 Sodium Xylenesulfonate, NTP TR 464 

IMPACT OF HELICOBACTER HEPATICUS

INFECTION IN B6C3F1 MICE

FROM 12 NTP 2-YEAR CARCINOGENESIS STUDIES

ABSTRACT

Male and female B6C3F1 mice from 12 NTP 2-year carcinogenesis studies were found to be infected with

Helicobacter hepaticus. Many of the male mice from nine of these studies (“affected” studies) had an

associated hepatitis. The current evaluations were performed in an attempt to determine if the data from the

H. hepaticus-affected NTP B6C3F1 mouse studies were compromised and unsuitable for cancer hazard

identification. The incidences of neoplasms of the liver (both hepatocellular neoplasms and

hemangiosarcoma), but not of other organs in control male B6C3F1 mice, were found to be increased in

affected studies compared to control males from unaffected studies. The increased incidence of

hepatocellular neoplasms was observed in those males exhibiting H. hepaticus-associated hepatitis. Other

observations further differentiated control male mice from affected and unaffected studies. H-ras codon 61 

CAA-to-AAA mutations were less common in liver neoplasms in males from affected studies compared to

historical and unaffected study controls. In addition, increases in cell proliferation rates and apoptosis were

observed in the livers of male mice with H. hepaticus-associated hepatitis. These data support the

hypothesis that the increased incidence of liver neoplasms is associated with H. hepaticus and that hepatitis

may be important in the pathogenesis. Therefore, interpretation of carcinogenic effects in the liver of

B6C3F mice may be confounded if there is H. hepaticus-associated hepatitis. 1 

INTRODUCTION

Helicobacter-Induced Diseases

Since the bacterium H. pylori was isolated from humans in 1983, numerous Helicobacter species have been

identified in several laboratory and domestic animal species. Their pathogenicity varies, with some species

inducing significant disease while others appear merely to colonize the gastrointestinal tract. H. pylori is

known to cause chronic gastritis and peptic ulcers in humans (Marshall and Warren, 1984; Graham, 1989; 

Lee et al., 1993) and, more recently, has been linked to adenocarcinoma and mucosa-associated lymphoma

of the stomach (Fox et al., 1989; Nomura et al., 1991; Parsonnet et al., 1991; Wotherspoon et al., 1993). 

Based on epidemiological and pathology findings, the International Agency for Research on Cancer (1994) 

has classified H. pylori as a group 1 carcinogen in humans. H. hepaticus is associated with an increase in

liver neoplasm incidences in A/JCr mice (Ward et al., 1994a; Fox et al., 1996). 

H. hepaticus commonly colonizes the gastrointestinal tract of many strains of mice from many sources (Fox

et al., 1994; Ward et al., 1994b; Shames et al., 1995). It has been shown to be pathogenic, with hepatitis

highly prevalent in some strains of mice (A/JCr, BALB/cAnNCr, C3H/HeNCr, SJL/NCr, and SCID/NCr) 

(Ward et al., 1994b). Intestinal colonization does not necessarily result in subsequent hepatitis, and the

conditions that lead to migration of the organism from the intestine to the liver have not been determined. 

H. hepaticus appears to reside primarily within the bile canaliculi. Male mice were reported to have a

greater incidence and severity of hepatitis than female mice, and this finding occurred in NTP studies as

well. The recently identified H. bilis, like H. hepaticus, colonizes the biliary tract, liver, and intestine of

mice. While H. bilis has been identified in animals with chronic hepatitis, whether it caused the hepatitis is

not known (Fox et al., 1995). 

The pathogenesis of H. hepaticus-induced disease has not been fully characterized. In susceptible strains of

mice, H. hepaticus can cause acute, focal, nonsuppurative, necrotizing hepatitis, which progresses to

chronic, active hepatitis characterized by minimal necrosis, hepatocytomegaly, oval cell hyperplasia, and
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 253 

cholangitis. H. hepaticus has been found to possess high levels of urease (Fox et al., 1994). H. hepaticus

is often isolated from the cecum and colon but is not necessarily isolated from the liver of A/JCr mice, even

though these animals develop severe hepatitis. Culture supernatants from several strains of H. hepaticus

and several other Helicobacter species were shown to cause cytopathic effects in a rodent hepatocyte cell

line (Taylor et al., 1995). Ward et al. (1996) suggested that autoimmunity may play a role in the

progressive hepatitis and carcinogenesis in livers infected with H. hepaticus. 

NTP Infectious Disease Surveillance

In 1993, during the histological evaluation of an NTP 2-year study, pathologists identified a constellation of

liver lesions (hepatitis) in control and treated male mice that was consistent with what would later be

described in mice infected with H. hepaticus (Ward et al., 1993, 1994a; Fox et al., 1994). Subsequently, 

pathology results from all mouse studies begun since 1984 (67 two-year studies) were reviewed for

diagnoses of the characteristic hepatitis; the lesions were identified in nine studies (NTP, 1998a,b,c,d,e,f). 

Silver stains revealed helical bacteria consistent with Helicobacter present in the liver of male mice in the

nine studies. 

Every reasonable measure is taken to prevent the occurrence of infectious diseases during NTP 2-year

carcinogenicity studies. When infections occasionally occur, care is taken to identify the causal agent and

its source, measures are taken to ensure that animals in later studies will not be infected, and the potential

impact on biological parameters (primarily neoplastic endpoints) important in interpretation of the study is

determined. To date, animals (control and treated) from a few studies have had a mild pulmonary

inflammatory response presumed to be caused by an infectious agent. In other studies, there have been

utero-ovarian infections with Klebsiella sp. (Rao et al., 1987) and fungal infections of the nasal cavity. For

scientifically valid reasons, interpretation of chemical-related effects was not considered significantly

compromised in any of these studies. Unlike the previous infections, H. hepaticus involves the liver, the

major metabolic organ, and has been associated with an increase in incidences of liver neoplasms in the

A/JCr mouse (Ward et al., 1994a). Therefore, when the contemporary epizootic of H. hepaticus infection

in the United States affected several NTP studies, use of the data for hazard identification was questioned. 

The first step was to determine the extent of the infection within NTP studies and then evaluate the impact

the infection had on biological parameters important in interpretation of the carcinogenic potential of test

chemicals. 

MATERIALS AND METHODS

Histologic Examination

Studies in which mice were potentially infected with H. hepaticus were identified by reviewing the summary

pathology tables for characteristic diagnoses: oval and/or biliary epithelial hyperplasia, hepatocyte

enlargement (often diagnosed as karyomegaly), chronic inflammation, and regenerative hyperplasia. All

13-week and 2-year studies begun by the NTP since 1984 and for which complete pathology data were

available (67 two-year studies) were examined. Eight contemporary studies in which the characteristic

lesions were not identified from pathology tables were randomly selected for histologic reevaluation. Slides

containing sections of hematoxylin- and eosin-stained livers from 20 to 25 control and 20 to 25 high-dose

male mice from each of seven 2-year studies and one 13-week study (10 animals from each group) were

reexamined microscopically for the presence of hepatitis potentially related to H. hepaticus infection. 

Hepatitis consistent with that observed with H. hepaticus infection was not observed in any of these studies. 

Liver sections from five or more animals from each of nine 2-year studies in which hepatitis was observed

were prepared using the Warthin-Starry silver stain or Steiner’s modification to identify silver-positive

helical bacteria. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

254 Sodium Xylenesulfonate, NTP TR 464 

PCR-RFLP Detection of Helicobacter DNA

Assays based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were

conducted at the NIEHS (Malarkey et al., 1997) and the University of Missouri Research Animal

Diagnostic and Investigative Laboratory (MU-RADIL) (Riley et al., 1996) on liver tissue from

approximately 20 animals from each of 32 NTP 2-year studies (including the nine affected studies) and three

NTP 13-week studies. The majority of these studies were selected because they were begun at

approximately the same time (1988-1990) as the nine affected studies. Also, two earlier studies (1984-

1985; mouse life-span and p-nitroaniline studies) and one later study (1993; methyleugenol) were selected. 

The mouse life-span study was designed to evaluate the incidences of spontaneous changes associated with

age; therefore, there is no NTP Technical Report. Pathology peer review is not complete for the

methyleugenol study, and the NTP Technical Report (NTP, 1998g) has not been completed. Frozen tissue

was available from 22 of these studies, while only formalin-fixed tissue was available for the remaining ten

2-year studies and the three 13-week studies. Most of the assays were conducted by MU-RADIL, which

used Helicobacter genus-specific primers; MU-RADIL used restriction endonucleases on a subset of

positives to determine if the species was H. hepaticus. DNA was isolated from frozen liver samples with a

QIAamp Tissue Kit (Qiagen Inc., Chatsworth, CA) according to the manufacturer’s recommendations or

routine phenol/chloroform extraction (Malarkey et al., 1997). DNA content and purity were determined

spectrophotometrically by measuring the A260/A280 optical density ratio. To isolate DNA from paraffinembedded samples, five 10-μm sections were washed twice with 1 mL xylene and twice with 500 μL

ethanol. Tissues were then dried within a vacuum centrifuge prior to DNA isolation as described above. 

Routine measures were taken to avoid contamination at every step from tissue collection to PCR

amplification, and concurrently run controls without DNA were consistently negative. 

Statistical Analyses

Multiple regression procedures were used to compare control neoplasm rates in the nine affected studies

with the 26 unaffected contemporary studies which had no histologic evidence of H. hepaticus-associated

liver disease. While frozen liver tissue was unavailable from 13 of these 26 studies, none showed the

hepatitis indicative of H. hepaticus and thus were assumed to be unaffected. Potential confounding factors

such as body weight, date study was begun, route of administration, and animal supplier were included as

covariables in the statistical analysis. 

Analysis for H-ras Codon 61 CAA-to-AAA Mutations

For analyses of formalin-fixed tissue, three to five unstained serial sections (10 μm thick) were cut from

paraffin blocks containing hepatocellular adenomas or carcinomas. Paraffin-embedded tissues were

deparaffinized and rehydrated prior to being digested with proteinase k overnight at 55E C to isolate DNA. 

Frozen tissues were digested with 10 mg/mL pronase in 1% sodium dodecyl sulfate in TNE buffer (10 mM

TRIS, 150 mM NaCl, and 2 mM EDTA; pH 7.5) overnight at 37E C; DNA was isolated by phenol

chloroform extraction and precipitated with ethanol (Marmur, 1961; Sills et al., 1995). 

Nested primers were used for amplification of exon 2 of H-ras by PCR. The outer primers were 5'-CCA

CTA AGC CTG TTG TGT TTT GCA G-3' (forward primer) and 5'-CTG TAC TGA TGG ATG TCC

TCG AAG GA-3' (reverse primer). The inner primers (second round of amplification) were 5'-GAC ATC

TTA GAC ACA GCA GTT-3' (forward primer) and 5'-GGT GTT GTT GAT GGC AAA TAC-3' (reverse

primer). Although the normal sequence of codon 60 is GCT, the forward PCR primer is made with a T at

the penultimate 3' base to create the restriction site for Mse1. 

A nonradioactive RFLP method was employed to identify CAA-to-AAA mutations in the H-ras gene at

codon 61 in liver neoplasms (Lee and Drinkwater, 1995). This was based on Mse1 enzyme restriction

cutting only the sequence 5'-TTAA-3'. Thus, Mse1 will detect CŁA conversion mutation at the first

position of codon 61. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 255 

Analysis of PCNA and Apoptosis

Detailed methods are included in a report by Nyska et al. (1997). Cell proliferation was assessed in

nonneoplastic areas of the liver, kidney, and lung by determining a PCNA S-phase labeling index (the

percentage of cells in S phase). The identification of apoptotic cells was based on morphologic criteria

(Garewal et al., 1996; Goldsworthy et al., 1996) and confirmed immunohistochemically by the terminal

deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) procedure (Gavrieli et al., 

1992). 

RESULTS AND DISCUSSION

Identification of H. hepaticus Infection in NTP Studies

Determining the extent of H. hepaticus infection involved a three-pronged approach of histologic evaluation, 

silver stains, and PCR-RFLP based assays; all were necessary because of the limitations identified for each. 

In NTP studies, and as reported in other studies (Ward et al., 1994b), there were no obvious clinical signs

of infection, and the only significant histologic lesion (hepatitis) was observed in the liver, primarily in

males. Therefore, summary pathology tables were reviewed to identify studies that may have been affected

by H. hepaticus-associated hepatitis. Male mice from nine studies were identified (Table L1) as having the

hepatitis. Eight of the nine studies were begun during a time span of about 6 months (July 1990 to

January 1991), while the other study was begun much earlier (October 1988). The hepatitis was not

observed in any 13-week studies. Use of histologic evaluation for identification of infected animals has

limitations, however. It is somewhat insensitive, as H. hepaticus has been cultured and identified by PCRRFLP methods within livers of animals with no histological evidence of infection (Fox et al., 1998). This

may be explained in part by the limited sampling (two liver sections) and the sometimes focal nature of

H. hepaticus-associated hepatitis. Also, while in the more severely affected animals the hepatitis appears

somewhat characteristic, component lesions of the hepatitis are not pathognomonic, and, when the hepatitis

is subtle in 2-year old animals, it is more difficult to recognize or attribute to H. hepaticus. 

Within affected studies, the incidences of the hepatitis in male mice varied from 16% to 78% (Table L1). 

While generally mild to moderate, the hepatitis varied in severity from barely detectable in some animals to

extensive liver involvement and regeneration in others. Only a few females were identified as having the

characteristic hepatitis (Table L1). In general, the incidences and severities of H. hepaticus-associated

hepatitis were similar between control and treated groups. This constellation of nonneoplastic liver lesions, 

while not pathognomonic, was certainly suggestive of an H. hepaticus infection, particularly when observed

in control animals. Characteristic lesions included proliferation of oval and/or biliary epithelial cells, 

hepatocyte enlargement (diagnosed as karyomegaly), and chronic inflammation. In many instances, areas of

regenerative hyperplasia were identified within diseased liver. 

Helicobacter spp. are not usually observed on routine histologic examination of hematoxylin and eosinstained sections of liver. The methods for confirmation of infection with Helicobacter include WarthinStarry silver stain or Steiner’s modification (Garvey et al., 1985) of this stain for direct microscopic

observation of the organisms in tissue; however, this can be a relatively insensitive technique when few

organisms are present. In most instances, histologic differentiation between Helicobacter species is not

possible. Speciation can usually be accomplished with electron microscopy, but this technique is both time

consuming and labor intensive. Microbiologic culture of feces, cecal smears, and fresh or frozen liver is

also possible. Currently, assays involving amplification of the DNA of the organism using PCR are the

most rapid and perhaps the most sensitive methods of detection, and the use of restriction endonucleases has

allowed a determination of the species present. PCR-based methods also can be used on feces, cecal

contents, or liver homogenates and are most sensitive when using fresh or frozen tissue (Riley et al., 1996; 

Malarkey et al., 1997). 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

256 Sodium Xylenesulfonate, NTP TR 464 

Using Warthin-Starry silver stains or Steiner’s modification on the livers of five or more animals per study, 

helical bacteria (Helicobacter) were identified in animals from the nine affected studies. In some animals, 

helical bacteria were numerous, suggesting a heavy bacterial burden in these infected animals. However, 

even in these animals with abundant organisms, few to none were observed in proliferative hepatic lesions

such as foci and neoplasms. Helical bacteria were not identified in approximately 25% of males with

moderate hepatitis and were rarely identified in males without hepatitis or in females. The absence of

identification of helical organisms by silver stains does not preclude infection, nor does the presence of

organisms confirm H. hepaticus. Based upon current knowledge, however, the characteristic liver lesions

in B6C3F mice, coupled with the presence of silver-positive helical organisms, are highly suggestive of 1 

H. hepaticus infection. 

As the NTP evaluation evolved, PCR-based assays were developed that appeared more sensitive than

histologic evaluation and silver stains for identification and speciation of Helicobacter. Therefore, 

PCR-RFLP-based assays were used to confirm the presence of pathogenic Helicobacter (primarily

H. hepaticus) within the nine affected studies and to determine whether there was H. hepaticus infection in

other NTP studies. Unfortunately, none of the PCR-based assays had been specifically developed for, or

proven reliable for use with, formalin-fixed tissue. Frozen tissue was available from a limited number of

animals from a limited number of NTP studies, including only three of the nine affected studies. 

Furthermore, available frozen liver was almost always limited to tissue from a neoplasm, and, based upon

results obtained with silver stains, organisms are generally not readily observed within proliferative hepatic

lesions, even when organisms are abundant in adjacent liver tissue. Because the availability of frozen tissue

was limited, a PCR-RFLP-based assay was developed and evaluated (Malarkey et al., 1997) for use with

frozen or formalin-fixed tissue. 

The NIEHS and MU-RADIL laboratories conducted PCR-RFLP-based assays on 32 NTP 2-year studies and

three NTP 13-week studies (data not shown); frozen tissues from 22 of the 2-year studies were available. 

All three bioassays in which hepatitis was identified and for which frozen tissue was available were positive

for H. hepaticus by the PCR-RFLP-based assays (Table L2). At a third laboratory, H. hepaticus was also

cultured from the liver tissue of animals in one of these studies (Fox et al., 1998). Formalin-fixed tissues

from two of the three studies were evaluated and were also positive; these tissues had been fixed in formalin

for less than 48 hours. In the other six affected studies, for which only formalin-fixed tissue was available, 

H. hepaticus was identified in only 1 of 120 animals (Table L2). This decreased sensitivity was considered

to be related to the prolonged formalin fixation (Malarkey et al., 1997) rather than proof of an absence of

H. hepaticus. The presence or absence of H. hepaticus apparently cannot be confirmed with current

PCR-RFLP-based assays in liver that has been fixed in formalin for long periods (weeks or months). In the

three 13-week studies with formalin-fixed tissue, only 1 of 30 animals was positive for H. hepaticus. 

Within the three affected, PCR-RFLP-positive 2-year studies, H. hepaticus was often identified by PCR in

frozen livers of mice that had no overt hepatitis. In fact, based upon the combined data from two studies

(including PCR results from three laboratories), of 57 animals without characteristic liver lesions, 13 of

24 male mice (54%) and 17 of 33 female mice (52%) were positive for H. hepaticus. Furthermore, 

H. hepaticus was identified by PCR in frozen liver of several animals from three “unaffected” studies in

which hepatitis typical of that associated with H. hepaticus was not observed (Table L2). Apparent

variability occurs between various strains of mice and between individual mice from affected studies in

developing hepatitis in response to H. hepaticus infection. One would assume that, within affected studies, 

most or all animals have been exposed to the organism, and even animals resistant to developing hepatitis

may have organisms within the liver. This assumption is supported by the fact that animals without hepatitis

are often positive with PCR-RFLP-based assays. Therefore, although alternative explanations are possible, 

the three PCR-RFLP-positive studies in which liver lesions are absent are assumed to be true positives. In

fact, helical organisms were identified with a silver stain in one animal from one of these studies (Malarkey

et al., 1997). Therefore, in addition to assessing the affect of H. hepaticus in the nine affected 2-year
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 257 

studies, the significance of a positive PCR-RFLP assay for H. hepaticus in the absence of liver lesions is

also an important question. 

Inconsistent Results with PCR-Based Methods

As with any technique, the PCR-RFLP-based assays have limitations even when used to assay fresh and

frozen tissue. One assessment of the variability in results of PCR and serologic analyses for Helicobacter

among three commercial laboratories revealed significant inconsistencies (Dew et al., 1997). Others

(J.M. Ward and J. Thigpen, personal communications) have obtained similarly inconsistent results when

sending replicate samples to different laboratories. Though the number of samples evaluated by both the

NIEHS and MU-RADIL laboratories was limited, there was good, but not complete, correlation of

PCR-RFLP results. Also, within the affected studies, the PCR assays were not positive in some animals

with liver disease. This result may be explained, in part, by the fact that the only frozen tissues available

were neoplasms; as described above, neoplasms are expected to have fewer organisms. 

Analysis of H. hepaticus-Affected and Unaffected Studies

for Incidence of Common Neoplasms

To determine whether the incidences of various neoplasms were different between control groups from

affected and unaffected studies, the nine affected studies were compared to 26 unaffected studies begun at

relatively similar times (Table L3). There were no statistically significant differences in body weight or

survival among the affected and unaffected studies. The neoplasms evaluated represent those that occurred

at high enough incidences in various organs for statistically significant differences to be detected. Using

multiple regression procedures, male mice in the nine affected studies were demonstrated to have a

significantly (P<0.05) increased incidence of only two neoplasm types, both of which were in the liver

(hepatocellular neoplasms and hemangiosarcoma), when compared to the unaffected studies. Because of

these differences, there was also a corresponding significant difference in the overall incidence of malignant

neoplasms (all sites) as well as in the overall proportion of neoplasm-bearing animals. No other tissue site

showed a significant difference in the incidence of neoplasms. For female mice, the slightly increased

incidence of hepatocellular neoplasms observed in the affected studies was not statistically significant. 

This seemingly simple analysis is complicated by several potential confounding variables. There have been

coordinate, time-related increases in body weight and in the incidence of liver neoplasms in mice in NTP

studies (Haseman, 1992). Table L4 presents the liver neoplasm incidences in relation to the dates the

studies began and clearly shows the increases in liver neoplasm incidences and body weights (Seilkop, 

1995). In assessing differences in neoplasm incidences between H. hepaticus-affected and unaffected

studies, the most relevant comparison would be between studies begun at approximately the same time. The

starts of 20 of the 26 unaffected studies were clustered near the early part of the time frame (April 1988 to

June 1990), while the starts of the affected studies were clustered toward the later end, with eight of the

nine studies begun between July 1990 and January 1991; incidences of liver neoplasms in these later studies

are expected to be higher based on trends in body weight alone. While the slightly increased incidences of

liver neoplasms observed in female control mice in the nine affected studies is likely due to clustering in

time, clearly, this alone cannot account for the increased liver neoplasm incidences observed in control male

mice in the affected studies (Table L3). 

Ideally, unaffected studies used in the above comparison should not only be free of histologic evidence of

infection with H. hepaticus but should be confirmed as negative by PCR assays. Thirteen of these

26 studies could not be confirmed as negative by PCR because frozen tissue was not available; however, 

H. hepaticus-associated hepatitis was not present in any of the 26 studies. Because these and other data

reported to date suggest that hepatitis is associated with neoplasm development in the liver, it seems

reasonable to include those 13 studies, unconfirmed by PCR, in this analysis. The majority of the

13 studies confirmed as negative by PCR were begun much earlier than the clearly affected studies, and, 

therefore, comparing them alone to the nine affected studies is not reasonable. Although not presented
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

258 Sodium Xylenesulfonate, NTP TR 464 

here, a number of comparisons were made with various groupings of studies based on the degree of

confidence in their infection status. Although the outcomes of the various comparisons varied somewhat, 

incidences of hepatocellular neoplasms and hemangiosarcomas of the liver were consistently increased in

control male mice from affected studies compared to control males from unaffected studies. Significantly

increased liver neoplasm incidences generally were not observed in females. Importantly, the following

data corroborate the findings and association with H. hepaticus identified in these analyses. 

Analysis of Hepatitis-Positive and Hepatitis-Negative Mice

for Liver Neoplasm Incidence

Several infectious agents known to be associated with increased incidences of neoplasms cause chronic

inflammation in the target tissue or organ. It is commonly hypothesized that this inflammatory process may

cause or contribute to the development of neoplasms. One approach to address this was to stratify the mice

from the affected studies according to the severity of hepatitis and examine liver neoplasm incidences in

relation to these groupings. Thus, animals within the nine affected studies were placed into three groups: 

1) animals with mild to moderate hepatitis considered related to H. hepaticus infection (+), 2) animals with

minimal to mild hepatitis that may have been associated with H. hepaticus (±), and 3) animals with no

hepatitis that was considered to be associated with H. hepaticus (S). Within these groupings, the incidence

of liver neoplasms was significantly increased (P<0.05) in males with mild to moderate H. hepaticusassociated hepatitis (+) when compared to animals without such hepatitis (Table L5). The neoplasm

incidence in animals with minimal lesions (±) was also increased. The liver neoplasm incidence in males

without hepatitis (58%) was similar to the incidence (54.8%) in males from the 26 unaffected studies

(Table L3). This analysis clearly suggests an association of H. hepaticus-associated hepatitis with increased

liver neoplasm incidences. Females showed a similar trend, albeit not significant; however, these

comparisons are weak because of the low numbers of females with hepatitis. 

Analysis of H-ras Oncogene Mutations in Liver Neoplasms in Mice

from Affected and Unaffected Studies

Liver neoplasms commonly occur in control B6C3F mice in 2-year studies. In the historical database of 1 

333 male and female mice with liver neoplasms, 106 (32%) had H-ras codon 61 CAA-to-AAA mutations

(Maronpot et al., 1995). This historical control database is composed primarily of male data; however, 

adequate numbers of females have been assayed, and there was no significant difference in the incidences of

CAA-to-AAA mutations between males and females. 

In an attempt to examine further whether H. hepaticus infection had an effect on the development of

hepatocellular neoplasms, neoplasms from control male mice from selected affected (NTP, 1998a,b,c) and

unaffected (NTP, 1993, 1998h) studies were evaluated for H-ras codon 61 CAA-to-AAA mutations

(Table L6). Only 6% (2/33) of the hepatocellular neoplasms from control males with hepatitis from three

affected studies had this mutation. This percentage is significantly (P<0.01) less than the 32% (11/34) 

observed in males from the two unaffected studies and less than the 32% (106/333) that occurred in

historical control animals. In addition, neoplasms from males without hepatitis from the affected, PCRpositive triethanolamine study (NTP, 1998a) and the unaffected, PCR-positive methyleugenol study (NTP, 

1998g) were evaluated; the incidences of mutations in those groups were 3/14 (21%) and 2/17 (12%), 

respectively. 

Neoplasms from control female mice (none had hepatitis) from affected and unaffected studies were

evaluated for the CAA-to-AAA mutation (Table L6). The mutation rate was low in both the affected studies

(1/25; 4%) and the unaffected study (1/11; 9%) when compared to the 32% observed in the historical

control groups. 

The finding of a different H-ras mutation profile in neoplasms of male mice from affected studies tends to

support the association of increased neoplasm incidences with H. hepaticus, although there is no mechanistic
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 259 

understanding behind this observation. In a study of H. hepaticus-infected A/JCr mice, ras mutations were

not detected in the 25 hepatocellular neoplasms analyzed using a PCR/single-strand conformation

polymorphism assay (Sipowicz et al., 1997). Because of the low spontaneous rate of liver neoplasms in the

A/JCr mouse, there are few or no conclusive data on ras mutations in uninfected animals, however. Point

mutations at codons 12, 13, and 61 of the Ki-, Ha- and N-ras genes were not identified in 45 early gastric

carcinomas in humans, whether or not H. pylori was present (Craanen et al., 1995). If the increased

incidence of hepatocellular neoplasms is associated with hepatitis, as many suspect, then one would expect

the neoplasms from animals without hepatitis to have a similar mutational profile as that of the historical

controls. The data do not provide a clear answer, because the hepatitis-free males from the affected

triethanolamine study (NTP, 1998a) and the males from the methyleugenol study (NTP, 1998g), which were

positive by PCR but lacked hepatitis, had mutation frequencies between those of the unaffected controls and

the hepatitis-positive mice. Furthermore, mutations in neoplasms from females, none of which had

hepatitis, from two affected and one unaffected study were very low compared to the historical controls. 

These findings were unexpected, and their significance is not understood. 

H. hepaticus-Associated Alterations in Cell Kinetics

Studies evaluating cell kinetics were completed to explore further the link between hepatitis and the

increased incidence of liver neoplasms (Table L7; Nyska et al., 1997). One of the major objectives was to

determine whether there were differences between PCNA labeling indices in the livers of animals with

hepatitis from three affected studies, cobalt sulfate heptahydrate, chloroprene, and triethanolamine (NTP, 

1998a,b,c), compared to animals without hepatitis, whether from the same three affected studies or from an

unaffected study, 1-trans-delta9

-tetrahydrocannabinol (NTP, 1996). Male mice with hepatitis from the three

affected studies had a significantly increased (P<0.001) labeling index, with a 24-fold increase over males

from the unaffected study and a sixfold increase over males without hepatitis from the same three affected

studies (Table L7). The labeling index increase in these mice was substantial and was considered

biologically significant. Male mice without hepatitis from the three affected studies had a significantly

greater labeling index (increased fourfold) than male mice from the unaffected study (Table L7). The

significance of this finding is uncertain, as differences of a similar magnitude were observed in other

comparisons. For example, the labeling index of females from the unaffected 1-trans-delta 9

-

tetrahydrocannabinol study (Table L7; NTP, 1996) was increased fivefold over females from the PCRpositive, hepatitis-negative scopolamine hydrobromide trihydrate study (NTP, 1997). Such differences may

be within the limits of normal variability for 2-year-old animals. 

A second objective of the cell proliferation studies of the liver was to determine if labeling indices were

increased in animals from the PCR-positive, hepatitis-negative methyleugenol (NTP, 1998g), scopolamine

hydrobromide trihydrate (NTP, 1997), and mouse life-span studies compared to an unaffected PCR-negative

and hepatitis-negative 1-trans-delta9

-tetrahydrocannabinol study (NTP, 1996). The scopolamine

hydrobromide trihydrate study was evaluated and included in the study by Nyska et al. (1997), while the

methyleugenol and mouse life-span studies were completed later and are included in Table L7. The labeling

indices of males from two of these three studies were almost identical to those of males from the unaffected

study. However, the labeling index of males from the mouse life-span study is increased approximately

fivefold over that of males from the unaffected study as well as fivefold over the labeling indices of males

from the two like studies of scopolamine hydrobromide trihydrate and methyleugenol. This finding suggests

that the increase observed in the mouse life-span study is not attributable to the presence of H. hepaticus, as

two other studies also positive for H. hepaticus did not show a similar increase. 

The cell proliferation data for the liver from NTP studies are consistent with data from a study by Fox et al. 

(1996) in which cell proliferation indices were evaluated at 8, 10, and 13 months in the A/JCr mouse, 

which is generally believed to be more susceptible to H. hepaticus-associated hepatitis than the B6C3F 1 

mouse. In the study by Fox et al. (1996), cell proliferation rates were significantly increased at all time

points in males. Some increases were observed in females in that study but did not reach statistical
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

260 Sodium Xylenesulfonate, NTP TR 464 

significance. An increased incidence of hepatocellular neoplasms was observed only in the males. Though

liver lesions were observed in females in that study, they were less severe than those in males. 

In addition to the liver, cell proliferation indices (PCNA) were evaluated in the kidneys and lungs of male

and female mice in affected studies versus those in unaffected studies (Nyska et al., 1997). No apparent

effect of H. hepaticus infection or the presence of hepatitis on PCNA indices was observed for the kidneys

or lungs. 

Apoptosis (programmed cell death) is another important parameter in evaluations of cell kinetics. The

apoptotic index in the liver of male mice with hepatitis from an affected study, cobalt sulfate heptahydrate

(NTP, 1998b), was significantly (P<0.01) greater than that observed in males from the unaffected 1-transdelta9

-tetrahydrocannabinol study and the PCR-positive, hepatitis-negative scopolamine hydrobromide

trihydrate study (Nyska et al., 1997). For females, there were no significant differences among the three

studies. 

Two 13-week studies which were begun during the same time as the nine affected studies were randomly

selected for evaluation of PCNA indices. H. hepaticus was not identified in either of the studies by

PCR-RFLP; however, as with all NTP 13-week studies, only tissue fixed in formalin for an unspecified

period was available. Because of this, no true negative control group was available; therefore, the labeling

index of these 19- to 20-week-old animals was compared to values cited in the literature (Eldridge and

Goldsworthy, 1996) for 20-week-old B6C3F mice. The labeling index in the NTP studies clearly was not 1 

increased (data not shown). 

The Impact of H. hepaticus on the Interpretation of 2-Year Carcinogenesis Studies

Increases in the incidences of neoplasms are associated with a number of infectious agents. The chronic

inflammation caused by these agents has been hypothesized to be important in the pathogenesis of the

increased neoplasm incidences (e.g., gastric cancer associated with H. pylori). The increased incidences of

liver neoplasms in male mice from the nine affected NTP studies were observed in the animals with

H. hepaticus-associated hepatitis. Neoplasms from males with hepatitis tended to have an H-ras mutation

profile different from that of animals from unaffected studies. Further, cell replication rates at 2 years were

significantly higher in males with hepatitis compared to those in males without hepatitis. The data suggest

that H. hepaticus-associated hepatitis is associated with the increased incidences of liver neoplasms in the

male B6C3F1 mouse. Therefore, the most important consideration in evaluating the impact of H. hepaticus

infection on the interpretation of study results appears to be the presence or absence of significant hepatitis. 

For any carcinogenicity study, data within and specific to the individual study provide the greatest basis for

an accurate interpretation. However, it is prudent to consider and evaluate all data or information which

may affect the interpretation. Based upon the data presented in this and other reports, general guidelines

emerge that may be useful in interpreting potential chemical-associated carcinogenic effects in H. hepaticusinfected B6C3F mice. In a study with sufficient evidence of H. hepaticus-associated hepatitis (>10% of 1 

the animals having the characteristic hepatitis may be a reasonable guideline), interpretation of increased

incidences of liver neoplasms (hepatocellular neoplasms and hemangiosarcoma) of male mice is considered

to be potentially confounded. 

Altered chemical uptake and metabolism, due to the intestinal load of H. hepaticus and to H. hepaticusassociated liver disease, respectively, are possible reasons for considering that the male mouse response to

chemical administration at sites other than the liver should also be considered confounded. Data do not

currently exist that definitively answer this question. In this group of nine studies, however, there is no

evidence to suggest that affected mice responded to chemical treatment in organs other than the liver in a

manner different from mice in nonaffected studies. Within each study, there was excellent concordance in

chemical-associated neoplasms between the male mice and the females, which had little or no hepatitis
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 261 

(Table L8). Furthermore, analyses indicate that H. hepaticus is not associated with neoplastic responses

outside the liver; incidences of neoplasms at sites other than the liver were not different between control

groups from affected and unaffected studies (Table L3). Cell replication rates in two major organs (lung

and kidney) also were not increased in control groups from affected studies compared to those from

unaffected studies. 

One of the more difficult issues to address is whether interpretation of a treatment-related increase in liver

neoplasm incidences in the female mouse is confounded when H. hepaticus-associated hepatitis is present

within the male mice in the study. Most evidence to date links hepatitis with the increased liver neoplasm

incidences observed in males, and female B6C3F mice in affected studies do not have significant hepatitis 1 

at 2 years. The lack of hepatitis in females, however, is based on an analysis in which only late time points

were evaluated histologically. Therefore, it is conceivable that hepatitis along with increased cell

proliferation could have occurred earlier and resolved by 18 months to 2 years. Data collected to date, 

however, suggest that H. hepaticus-associated hepatitis is a late-developing and persistent disease in the

B6C3F1 mouse. H. hepaticus-associated hepatitis has never been observed in any NTP 13-week studies, 

including five begun during the same 6-month time span as eight of the nine affected 2-year studies. Also, 

within affected 2-year studies, more males (51%) that were 18 to 24 months of age had hepatitis than those

(34%) that were 12 to 18 months of age. This is consistent with a report by Ward et al. (1994b) that

H. hepaticus-associated liver lesions are not observed at early time points in the B6C3F 1 mouse. 

Nonetheless, within affected studies, female control mice did have a slightly elevated incidence of liver

neoplasms when compared to control mice from unaffected studies, and the data derived from the H-ras

mutation frequency analysis were inconclusive. The possibility that H. hepaticus-infected female mice from

affected studies may respond differently to a liver carcinogen than mice from unaffected studies cannot be

eliminated at this time. However, because within an affected study hepatitis is observed only rarely in

females, until definitive data suggest otherwise, it is concluded that the interpretation of an apparent

chemical-induced neoplastic effect in the liver of female mice is not confounded. To censor the few females

with H. hepaticus-associated hepatitis from any statistical analyses of hepatocellular neoplasms would be

prudent. Studies in the ostensibly more sensitive A/JCr mouse (Fox et al., 1996) also showed significant

increases in neoplasm incidences and cell proliferation rates in the liver of H. hepaticus-infected males, but

not females. 

Another concern is how to interpret possible chemical-related effects in a study in which the status of

H. hepaticus infection cannot be determined by PCR-RFLP because only tissues fixed in formalin for more

than 48 hours are available. While histologic evaluation is inadequate to identify infection, it appears

adequate for identifying hepatitis severe enough to alter the outcome of the study. Therefore, in the absence

of significant histologic evidence of H. hepaticus-associated hepatitis, the outcome of a 2-year study should

not be considered potentially compromised. 

The causality between H. hepaticus infection and neoplasia has not been proven in the B6C3F 1 mouse in

these studies, nor has the mechanism of this association been determined; further studies are needed. 

However, sufficient information exists to make reasonable scientific judgments relative to the interpretation

of data from the nine 2-year carcinogenicity studies in the B6C3F1 mouse. Refinements to the above

interpretive positions may occur if warranted by future information. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

262 Sodium Xylenesulfonate, NTP TR 464 

REFERENCES

Craanen, M.E., Blok, P., Top, B., Boerrigter, L., Dekker, W., Offerhaus, G.J.A., Tytgat, G.N.J., and
 

Rodenhuis, S. (1995). Absence of ras gene mutations in early gastric carcinomas. Gut 37, 758-762.
#

Dew, J.A., Clifton, L.G., Sanders, B.L., and Reynolds, R.P. (1997). Comparison of results of
 

Helicobacter tests performed by commercial laboratories. Contemp. Top. (AALAS) 36, 60. (Abstr.)
#

Eldridge, S.R., and Goldsworthy, S.M. (1996). Cell proliferation rates in common cancer target tissues of
 

B6C3F1 mice and F344 rats: Effects of age, gender, and choice of marker. Fundam. Appl. Toxicol. 32,
#

159-167.
#

Fox, J.G., Correa, P., Taylor, N.S., Zavala, D., Fontham, E., Janney, F., Rodriquez, E., Hunter, F., and
 

Diavolitsis, S. (1989). Campylobacter pylori-associated gastritis and immune response in a population at
 

increased risk of gastric carcinoma. Am. J. Gastroenterol. 84, 775-781.
#

Fox, J.G., Dewhirst, F.E., Tully, J.G., Paster, B.J., Yan, L., Taylor, N.S., Collins, M.J., Jr.,
#

Gorelick, P.L., and Ward, J.M. (1994). Helicobacter hepaticus sp. nov., a microaerophilic bacterium
 

isolated from livers and intestinal mucosal scrapings from mice. J. Clin. Microbiol. 32, 1238-1245.
#

Fox, J.G., Yan, L.L., Dewhirst, F.E., Paster, B.J., Shames, B., Murphy, J.C., Hayward, A.,
#

Belcher, J.C., and Mendes, E.N. (1995). Helicobacter bilis sp. nov., a novel Helicobacter species isolated
 

from bile, livers, and intestines of aged, inbred mice. J. Clin. Microbiol. 33, 445-454.
#

Fox, J.G., Li, X., Yan, L., Cahill, R.J., Hurley, R., Lewis, R., and Murphy, J.C. (1996). Chronic
 

proliferative hepatitis in A/JCr mice associated with persistent Helicobacter hepaticus infection: A model of
 

Helicobacter-induced carcinogenesis. Infect. Immun. 64, 1548-1558.
#

Fox, J.G., MacGregor, J., Shen, Z., Li, X., Lewis, R., and Dangler, C.A. (1998). Role of Helicobacter
 

hepaticus in confounding results of a triethanolamine carcinogenesis study in mice. J. Clin. Microbiol.
#

(in press)
#

Garewal, H., Bernstein, H., Bernstein, C., Sampliner, R., and Payne, C. (1996). Reduced bile acidinduced apoptosis in “normal” colorectal mucosa: A potential biological marker for cancer risk. Cancer
 

Res. 56, 1480-1483.
#

Garvey, W., Fathi, A., and Bigelow, F. (1985). Modified Steiner for the demonstration of spirochetes. 

J. Histotechnol. 8, 15-17. 

Gavrieli, Y., Sherman, Y., and Ben-Sasson, S.A. (1992). Identification of programmed cell death in situ

via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119, 493-501. 

Goldsworthy, T.L., Fransson-Steen, R., and Maronpot, R.R. (1996). Importance of and approaches to

quantification of hepatocyte apoptosis. Toxicol. Pathol. 24, 24-35. 

Graham, D.Y. (1989). Campylobacter pylori and peptic ulcer disease. Gastroenterology 96, 615-625. 

Haseman, J.K. (1992). Value of historical controls in the interpretation of rodent tumor data. Drug Inf. J. 

26, 191-200. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 263 

International Agency for Research on Cancer (IARC) (1994). IARC Monographs on the Evaluation of

Carcinogenic Risks to Humans: Schistosomes, Liver Flukes and Helicobacter pylori, Vol. 61. IARC, 

Lyon, France. 

Lee, A., Fox, J., and Hazell, S. (1993). Pathogenicity of Helicobacter pylori: A perspective. Infect. 

Immun. 61, 1601-1610. 

Lee, G.-H., and Drinkwater, N.R. (1995). Hepatocarcinogenesis in BXH recombinant inbred strains of

mice: Analysis of diverse phenotypic effects of the hepatocarcinogen sensitivity loci. Mol. Carcinog. 14, 

190-197. 

Malarkey, D.E., Ton, T.-V., Hailey, J.R., and Devereaux, T.R. (1997). A PCR-RFLP method for the

detection of Helicobacter hepaticus in frozen or fixed liver from B6C3F mice. Toxicol. Pathol. 25, 1 

606-612. 

Marmur, J. (1961). A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J. Mol. 

Biol. 3, 208-218. 

Maronpot, R.R., Fox, T., Malarkey, D.E., and Goldsworthy, T.L. (1995). Mutations in the ras protooncogene: Clues to etiology and molecular pathogenesis of mouse liver tumors. Toxicology 101, 125-156. 

Marshall, B.J., and Warren, J.R. (1984). Unidentified curved bacilli in the stomach of patients with

gastritis and peptic ulceration. Lancet 1, 1311-1314. 

National Toxicology Program (NTP) (1993). Toxicology and Carcinogenesis Studies of Oxazepam (CAS

No. 604-75-1) in Swiss Webster and B6C3F1 Mice (Feed Studies). Technical Report Series No. 443. NIH

Publication No. 93-3359. U.S. Department of Health and Human Services, Public Health Service, National

Institutes of Health, Research Triangle Park, NC. 

National Toxicology Program (NTP) (1996). Toxicology and Carcinogenesis Studies of 1-Trans-delta 9

-

tetrahydrocannabinol (CAS No. 1972-08-3) in F344/N Rats and B6C3F Mice (Gavage Studies). Technical 1 

Report Series No. 446. NIH Publication No. 97-3362. U.S. Department of Health and Human Services, 

Public Health Service, National Institutes of Health, Research Triangle Park, NC. 

National Toxicology Program (NTP) (1997). Toxicology and Carcinogenesis Studies of Scopolamine

Hydrobromide Trihydrate (CAS No. 6533-68-2) in F344/N Rats and B6C3F Mice (Gavage Studies). 1 

Technical Report Series No. 445. NIH Publication No. 97-3361. U.S. Department of Health and Human

Services, Public Health Service, National Institutes of Health, Research Triangle Park, NC. 

National Toxicology Program (NTP) (1998a). Toxicology and Carcinogenesis Studies of Triethanolamine

(CAS No. 102-71-6) in F344/N Rats and B6C3F Mice (Dermal Studies). Technical Report Series 1 

No. 449. NIH Publication No. 98-3365. U.S. Department of Health and Human Services, Public Health

Service, National Institutes of Health, Research Triangle Park, NC. (in preparation) 

National Toxicology Program (NTP) (1998b). Toxicology and Carcinogenesis Studies of Cobalt Sulfate

Heptahydrate (CAS No. 10026-24-1) in F344/N Rats and B6C3F Mice (Inhalation Studies). Technical 1 

Report Series No. 471. NIH Publication No. 98-3961. U.S. Department of Health and Human Services, 

Public Health Service, National Institutes of Health, Research Triangle Park, NC. (in press) 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

264 Sodium Xylenesulfonate, NTP TR 464 

National Toxicology Program (NTP) (1998c). Toxicology and Carcinogenesis Studies of Chloroprene

(CAS No. 126-99-8) in F344/N Rats and B6C3F Mice (Inhalation Studies). Technical Report Series 1 

No. 467. NIH Publication No. 98-3957. U.S. Department of Health and Human Services, Public Health

Service, National Institutes of Health, Research Triangle Park, NC. (in press) 

National Toxicology Program (NTP) (1998d). Toxicology and Carcinogenesis Studies of Technical Grade

Sodium Xylenesulfonate (CAS No. 1300-72-7) in F344/N Rats and B6C3F Mice (Dermal Studies). 1 

Technical Report Series No. 464. NIH Publication No. 98-3380. U.S. Department of Health and Human

Services, Public Health Service, National Institutes of Health, Research Triangle Park, NC. (in press) 

National Toxicology Program (NTP) (1998e). Toxicology and Carcinogenesis Studies of AZT (CAS

No. 30516-87-1) and AZT/"-Interferon A/D in B6C3F Mice (Gavage Studies). Technical Report Series 1 

No. 469. NIH Publication No. 98-3959. U.S. Department of Health and Human Services, Public Health

Service, National Institutes of Health, Research Triangle Park, NC. (in press) 

National Toxicology Program (NTP) (1998f). Toxicology and Carcinogenesis Studies of Theophylline

(CAS No. 58-55-9) in F344/N Rats and B6C3F Mice (Feed and Gavage Studies). Technical Report Series 1 

No. 473. NIH Publication No. 98-3963. U.S. Department of Health and Human Services, Public Health

Service, National Institutes of Health, Research Triangle Park, NC. (in press) 

National Toxicology Program (NTP) (1998g). Toxicology and Carcinogenesis Studies of Methyleugenol

(CAS No. 93-15-2) in F344/N Rats and B6C3F Mice (Gavage Studies). Technical Report Series No. 491. 1 

NIH Publication No. 98-3950. U.S. Department of Health and Human Services, Public Health Service, 

National Institutes of Health, Research Triangle Park, NC. (in preparation) 

National Toxicology Program (NTP) (1998h). Toxicology and Carcinogenesis Studies of Diethanolamine

(CAS No. 111-42-2) in F344/N Rats and B6C3F Mice (Dermal Studies). Technical Report Series 1 

No. 478. NIH Publication No. 98-3968. U.S. Department of Health and Human Services, Public Health

Service, National Institutes of Health, Research Triangle Park, NC. (in press) 

Nomura, A., Stemmermann, G.N., Chyou, P., Kato, I., Perez-Perez, G.I., and Blaser, M.J. (1991). 

Helicobacter pylori infection and gastric carcinoma among Japanese Americans in Hawaii. N. Engl. J. 

Med. 325, 1132-1136. 

Nyska, A., Maronpot, R.R., Eldridge, S.R., Haseman, J.K., and Hailey, J.R. (1997). Alteration in cell

kinetics in control B6C3F mice infected with Helicobacter hepaticus. Toxicol. Pathol. 25, 591-596. 1 

Parsonnet, J., Friedman, G.D., Vandersteen, D.P., Chang, Y., Vogelman, J.H., Orentreich, N., and

Sibley, R.K. (1991). Helicobacter pylori infection and the risk of gastric carcinoma. N. Engl. J. Med. 

325, 1127-1131. 

Rao, G.N., Hickman, R.L., Seilkop, S.K., and Boorman, G.A. (1987). Utero-ovarian infection in aged

B6C3F1 mice. Lab. Animal Sci. 37, 153-158. 

Riley, L.K., Franklin, C.L., Hook, R.R., Jr., and Besch-Williford, C. (1996). Identification of murine

Helicobacters by PCR and restriction enzyme analyses. J. Clin. Microbiol. 34, 942-946. 

Seilkop, S.K. (1995). The effect of body weight on tumor incidence and carcinogenicity testing in B6C3F 1 

mice and F344 rats. Fundam. Appl. Toxicol. 24, 247-259. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 265 

Shames, B., Fox, J.G., Dewhirst, F., Yan, L., Shen, Z., and Taylor, N.S. (1995). Identification of
 

widespread Helicobacter hepaticus infection in feces in commercial mouse colonies by culture and PCR
 

assay. J. Clin. Microbiol. 33, 2968-2972.
#

Sills, R.C., Hong, H.L., Greenwell, A., Herbert, R.A., Boorman, G.A., and Devereux, T.R. (1995). 

Increased frequency of K-ras mutations in lung neoplasms from female B6C3F1 mice exposed to ozone for
 

24 or 30 months. Carcinogenesis 16, 1623-1628.
#

Sipowicz, M.A., Weghorst, C.M., Shio, Y.-H., Buzard, G.S., Calvert, R.J., Anver, M.R.,
#

Anderson, L.M., and Rice, J.M. (1997). Lack of p53 and ras mutations in Helicobacter hepaticus-induced
 

liver tumors in A/JCr mice. Carcinogenesis 18, 233-236.
#

Taylor, N.S., Fox, J.G., and Yan, L. (1995). In-vitro hepatotoxic factor in Helicobacter hepaticus,
#

H. pylori and other Helicobacter species. J. Med. Microbiol. 42, 48-52. 

Ward, J.M., Anver, M.R., Haines, D.C., Tully, J.G., Jr., Collins, M.J., Jr., Gorelick, P.L., 

Anderson, L., Rice, J.M., and Russell, R.J. (1993). A unique hepatitis in mice associated with a helical

bacterium. Toxicol. Pathol. 21, 591. (Abstr.) 

Ward, J.M., Fox, J.G., Anver, M.R., Haines, D.C., George, C.V., Collins, M.J., Jr., Gorelick, P.L., 

Nagashima, K., Gonda, M.A., Gilden, R.V., Tully, J.G., Russell, R.J., Benveniste, R.E., Paster, B.J., 

Dewhirst, F.E., Donovan, J.C., Anderson, L.M., and Rice, J.M. (1994a). Chronic active hepatitis and

associated liver tumors in mice caused by a persistent bacterial infection with a novel Helicobacter species. 

J. Natl. Cancer Inst. 86, 1222-1227. 

Ward, J.M., Anver, M.R., Haines, D.C., and Benveniste, R.E. (1994b). Chronic active hepatitis in mice

caused by Helicobacter hepaticus. Am. J. Pathol. 145, 959-968. 

Ward, J.M., Benveniste, R.E., Fox, C.H., Battles, J.K., Gonda, M.A., and Tully, J.G. (1996). 

Autoimmunity in chronic active Helicobacter hepatitis of mice. Am. J. Pathol. 148, 509-517. 

Wotherspoon, A.C., Doglioni, C., Diss, T.C., Pan, L., Moschini, A., de Boni, M., and Isaacson, P.G. 

(1993). Regression of primary low-grade B-cell gastric lymphoma of mucosa-associated lymphoid tissue

type after eradication of Helicobacter pylori. Lancet 342, 575-577. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

266 Sodium Xylenesulfonate, NTP TR 464 

TABLE L1 

Incidence of Helicobacter hepaticus-Associated Hepatitis in Control B6C3F1 Mice

from Nine NTP 2-Year Studiesa

Incidence of Hepatitis (%) 

Study Males Females

Sodium xylenesulfonate 78 4 

AZT/5,000 U "-interferon A/D 76 4 

Cobalt sulfate heptahydrate 72 8 

AZT/500 U "-interferon A/D 66 0 

Chloroprene 54 0 

Theophylline 32 0 

"-Interferon A/D 22 4 

Triethanolamine 20 0 

AZT 16 2 

Average 48 2 

a Includes regeneration and mild to marked (excludes minimal) chronic inflammation, karyomegaly, oval cell hyperplasia, and bile duct

hyperplasia. AZT ̆3N-azido-3N-deoxythymidine

TABLE L2 

Identification of Helicobacter hepaticus with PCR-RFLP-Based Assays in Control B6C3F1 Mice

from 32 NTP 2-Year Studies and Three NTP 13-Week Studiesa

H. hepaticus-Positive Studiesb

Type of Sample Total Affected Unaffected

Studies Studies Studies

13-Week Studies

Formalin-fixed liver 3 — 1/3c

2-Year Studies

Frozen liver 22 3/3 3/19 

Formalin-fixed liver 10 1/6c 0/4 

a PCR-RFLP ̆polymerase chain reaction-restriction fragment length polymorphism b	 Number of H. hepaticus-positive studies/number of affected or unaffected studies. Affected studies are those in which hepatitis typical of

that associated with H. hepaticus infection occurred in many male mice. 

Only one animal in the positive study was positive for H. hepaticus. c
 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 267 

TABLE L3 

Comparison of Neoplasm Incidences in Control B6C3F1 Mice

from Helicobacter hepaticus-Affected and Unaffected NTP 2-Year Studies

Males Females

Affected Unaffected Affected Unaffected

Studiesa Studies Studies Studies

Number of studies 9 26 9 26 

Survival (%) 64 71 68 68 

12-Month body wt (g) 48.0 48.3 48.1 47.0 

Neoplasm incidence (%) 

Liver 71.3* 54.8 50.3 40.5 

Lung 26.6 23.2 7.6 10.3 

Pituitary gland 0.4 0.8 14.7 14.3 

Harderian gland 5.6 6.1 6.0 4.9 

Lymphoma 6.9 6.3 16.2 15.5 

Circulatory system 9.8 6.0 5.3 4.7 

liver only 7.1* 2.5 — —

All benign 61.8 57.2 59.1 54.6 

All malignant 61.3* 40.9 50.0 44.2 

All neoplasms 88.0* 77.4 82.7 75.4 

* Significantly different (P#0.05) from the unaffected studies a Affected studies are those in which hepatitis typical of that associated with H. hepaticus infection occurred in many male mice. 
 

 

 

 

 

 

 

 

268 Sodium Xylenesulfonate, NTP TR 464 

TABLE L4 

Liver Neoplasm Incidences and Body Weights of Control B6C3F1 Mice

in Relation to Study Start Dates of Helicobacter hepaticus-Affected and Unaffected NTP 2-Year Studiesa

Liver Neoplasm Incidence (%) Mean Body Weight (g) 

Study Start Date Affected Unaffected Affected Unaffected a Studies Studies Studies Studies

Male

April to September 1988 — 43.8 (8)b — 46.2 (8) 

October 1988 62.0 (1) — 48.3 (1) —

November 1988 to September 1989 — 52.6 (7) — 48.7 (7) 

October 1989 to June 1990 — 61.2 (5) — 48.9 (5) 

July 1990 to January 1991 72.5 (8) 66.2 (4) 48.0 (8) 49.0 (4) 

February 1991 to April 1992 — 68.0 (2) — 52.8 (2) 

Average 71.3 54.8 48.0 48.3 

Female

April to September 1988 — 31.1 (8) — 44.8 (8) 

October 1988 46.0 (1) — 46.4 (1) —

November 1988 to September 1989 — 39.9 (7) — 47.2 (7) 

October 1989 to June 1990 — 38.6 (5) — 45.9 (5) 

July 1990 to January 1991 50.9 (8) 54.2 (4) 48.3 (8) 48.0 (4) 

February 1991 to April 1992 — 58.0 (2) — 55.6 (2) 

Average 50.3 40.5 48.1 47.0 

a Includes nine affected studies (those in which hepatitis typical of that associated with H. hepaticus infection occurred in many male mice) and

26 unaffected studies

b Number of studies is given in parentheses. 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

Sodium Xylenesulfonate, NTP TR 464 269 

TABLE L5 

Association of Liver Neoplasm Incidence and Severity of Helicobacter hepaticus-Associated Hepatitis a in Control B6C3F Mice from Nine Affected NTP 2-Year Studies 1 

Liver Neoplasm Incidence

Severity of Hepatitis Males Females

Absent

Minimal

Mild/moderate

Significance of association

101/175 (58%) 

44/57 (77%) 

176/218 (81%) 

P<0.05 

196/396 (49%) 

23/42 (55%) 

7/11 (64%) 

NSb

a Affected studies are those in which hepatitis typical of that associated with H. hepaticus infection occurred in many male mice. b NS ̆not significant

TABLE L6 

H-ras Codon 61 AAA Mutations in Spontaneous Liver Neoplasms in Control B6C3F1 Mice

from Helicobacter hepaticus-Affected and Unaffected NTP 2-Year Studies

a Study Affected H-ras AAA Mutations

Male

Cobalt sulfate heptahydrate + 0/10 (0%) 

Chloroprene + 1/13 (8%) 

Triethanolamine + 1/10 (10%) 

Oxazepam — 7/18 (39%) 

Diethanolamine — 4/16 (25%) 

Historical control database 106/333 (32%) 

Female

Chloroprene + 0/10 (0%) 

Triethanolamine + 1/15 (7%) 

Diethanolamine — 1/11 (9%) 

Historical control database 106/333 (32%) 

a + ̆affected; —  ̆not affected. Affected studies are those in which hepatitis typical of that associated with H. hepaticus infection occurred in

many male mice. 
 

 

 

 

 

 

 

 

 

 

 

 

270 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE L7 a Proliferating Cell Nuclear Antigen Labeling Indices in the Liver of Control B6C3F1 Mice

Hepatitis No. of PCNA Labeling Average PCNA

Animals Indexb Labeling Indexc

Male

Cobalt sulfate heptahydrated

Chloroprened

Triethanolamined

+ 

+ 

+ 

15 

12 

9 

0.535 ± 0.129 

1.452 ± 0.386 

1.215 ± 0.374 1.011 

Cobalt sulfate heptahydrate

Chloroprene

Triethanolamine

—

—

—

7 

10 

12 

0.175 ± 0.117 

0.296 ± 0.124 

0.100 ± 0.042 0.186 

1-Trans-delta -tetrahydrocannabinol 9 e

Scopolamine hydrobromide trihydratef

Methyleugenolf

Mouse life-span studyf

—

—

—

—

15 

14 

14 

15 

0.042 ± 0.011 

0.043 ± 0.012 

0.077 ± 0.020 

0.217 ± 0.880 

Female

Cobalt sulfate heptahydrate + 5 0.161 ± 0.062 

Cobalt sulfate heptahydrate

Chloroprene

Triethanolamine

—

—

—

17 

12 

12 

0.055 ± 0.015 

0.154 ± 0.050 

0.138 ± 0.053 0.108 

1-Trans-delta -tetrahydrocannabinol 9 — 13 0.156 ± 0.047 

Scopolamine hydrobromide trihydrate — 15 0.032 ± 0.009 

a A portion of these data are presented in Nyska et al. (1997). + ̆hepatitis present; —  ̆no hepatitis present b Mean ± standard error; PCNA ̆proliferating cell nuclear antigen
 

Average of the mean labeling indices for animals from all three studies
 d	 Affected study (one in which hepatitis typical of that associated with H. hepaticus occurred in many male mice) e Unaffected study (one in which the typical hepatitis did not occur in mice) f	 Unaffected study with no typical hepatitis, but positive for H. hepaticus by polymerase chain reaction-restriction fragment length

polymorphism-based assay
 

 

 

 

 

 

271 

c

Sodium Xylenesulfonate, NTP TR 464 

TABLE L8 

Summary of Target Sites of Carcinogenicity in B6C3F Mice from NTP 2-Year Studies 1 

with Helicobacter hepaticus-Associated Hepatitis

Males	 Females

Chloroprene Lung Lung

Circulatory systema Circulatory system

Harderian gland Harderian gland

Forestomach Forestomach

Kidney Liver

Skin

Mesentery

Zymbal’s gland

Mammary gland

Cobalt sulfate heptahydrateb Lung Lung

Triethanolamine Liver Liver

AZTc None Vagina

Sodium xylenesulfonate None None

Theophylline None None

a Hemangioma and hemangiosarcoma of the liver were excluded from the analysis in males. b	 An apparent treatment-related increase in the incidence of hemangiosarcoma of the liver was discounted in male mice because of the presence

of H. hepaticus. 

AZT ̆3N-azido-3N-deoxythymidine. Includes four studies: AZT; "-interferon A/D; AZT/500 U "-interferon A/D; and

AZT/5,000 U "-interferon A/D
272 Sodium Xylenesulfonate, NTP TR 464