NATIONAL TOXICOLOGY PROGRAM 

Technical Report Series 

No. 445 

TOXICOLOGY AND CARCINOGENESIS 

STUDIES OF 

SCOPOLAMINE HYDROBROMIDE TRIHYDRATE 

(CAS NO.6533-68-2) 

IN F344/N RATSAND B6C3Fl MICE 

(GAVAGE STUDIES) 

US.DEPARTMENT OF HEALTHANDHUMANSERVICES 

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 aspectsof the chronic studies were subjected to retrospective quality assurance audits before 

being presented for public review. 

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

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

Chemicals selected for NTP toxicology and carcinogenesis studies are chosen primarily on the bases of human 

exposure, level of production, and chemical structure. The interpretive conclusions presented in this Technical 

Report are based only on the results of these NTP studies. Extrapolation of these results to other species, and 

quantitative risk analyses for humans, require wider analyses beyond the purview of these studies. Selection 

per se is not an indicator of a chemical’s carcinogenic potential. 

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

U.S. DepartmentofCommerce,5285PortRoyalRoad,Springfield,VA22161 (703487-4650). Single copies 

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

NIEHS, P.O. Box 12233, MD El-02, Research TrianglePark,NC27709(919-541-3419).Listingsofall 

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

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

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

ON THE 

TOXICOLOGY AND CARCINOGENESIS 

STUDIES OF 

SCOPOLAMINE HYDROBROMIDE TRIHYDRATE 

(CAS NO. 6533-68-2) 

IN F344/N RATS AND B6C3Fl MICE 

(GAVAGE STUDIES) 

NATIONAL TOXICOLOGY PROGRAM 

P.O. Box 12233 

ResearchTrianglePark, NC 27709 

March 1997 

NTP TR 445 

NIH Publication No. 97-3361 

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES 

Public Health Service 

National Institutes of Health 
2 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

CONTRIBUTORS 

National Toxicology Program 

Evaluated and interpreted results and reported findings 

K.M. Abdo, Ph.D., StudyScientist 

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

D.A. Bridge, B.S. 

J.R. Bucher, Ph.D. 

L.T. Burka, Ph.D. 

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

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

J.K. Haseman, Ph.D. 

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

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

J.H. Roycroft, Ph.D. 

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

D.B. Walters, Ph.D. 

K.L. Witt, M.S., Oak RidgeAssociatedUniversities 

Southern Research Institute 

Conducted studies, evaluated pathology findings for 16-day studies 

in rats and mice 

J.D. Prejean, Ph.D., PrincipalInvestigator 

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

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

Conducted studies, evaluated pathology findings for 14-week studies 

in rats and mice 

J.D. Prejean, Ph.D., PrincipalInvestigator 

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

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

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. Kurt~, Ph. D ., Principal Investigator 

S.D. Dannemiller, D.V.M. 

P.L. Jepsen, D.V.M. 

J.D. Johnson, Ph.D. 

M.J. Ryan, D.V.M, Ph.D. 

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

Experimental Pathology Laboratories, Inc. 

Provided pathology quality assurance 

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

S. Botts, D.V.M., Ph.D. 

B.F. Hamilton, D.V.M., Ph.D. 

Dynamac Corporation 

Prepared quality assurance audits 

S. Brecher, Ph.D., PrincipalInvestigator 

NTP Pathology Working Group 

Evaluated slides, prepared pathology report on rats 

(25June 1992) 

P.K. Hildebrandt, D.V.M., Chairperson 

PATHCO,Inc. 

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

National Toxicology Program 

B.F. Hamilton, D.V.M., Ph.D. 

Experimental Pathology Laboratories, Inc. 

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

NationalToxicologyProgram 

D. Mueten, D.V.M., Ph.D. 

North Carolina State University 

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

National Toxicology Program 

Evaluated slides, prepared pathology report on mice 

(8 December 1994) 

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

Pathology Associates, Inc. 

S. Botts, D.V.M., Ph.D. 

ExperimentalPathologyLaboratories,Inc. 

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

Chemical Industry Institute of Toxicology 

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

National Toxicology Program 

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

National Toxicology Program 

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

National Toxicology Program 

Analytical Sciences, Inc. 

Provided statistical analyses 

R.W. Morris, M.S., PrincipalInvestigator 

N.G. Mintz, B.S. 

S. Rosenblum, MS. 

Biotechnical Services, Inc. 

Prepared Technical Report 

S.R. Gunnels, M.A., PrincipalInvestigator 

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 ........... 10 

TECHNICAL REPORTS REVIEW SUBCOMMITTEE ............................. 11 

SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMI'ITEE COMMENTS ......... 12 

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

MATERIALSANDMETHODS ............................................ 21 

RESULTS .......................................................... 33 

DISCUSSION AND CONCLUSIONS ......................................... 57 

REFERENCES ........................................................ 61 

APPENDIX A Summary of Lesionsin Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate ........................... 67 

APPENDIX B Summary of LesionsinFemale Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate ........................... 109 

APPENDIXC Summary of Lesionsin Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate ........................... 139 

APPENDIX D Summary of Lesions in Female Micein the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate ........................... 181 

APpENDnr E GeneticToxicology ........................................... 217 

APPENDIX F Organ Weights and Organ-Weight-to-Body-Weight Ratios ................ 227 

APPENDIX G Hematology Results ......................................... 239 

APPENDIX H Reproductive Tissue Evaluations and Estrous Cycle Characterization ......... 245 

APPENDIX I Neurobehavioral Studies ...................................... 249 

Appendix J Chemical Characterization and Dose Formulation Studies ................. 257 

APPENDIX K Ingredients. Nutrient Composition. and Contaminant Levels 

in NIH-07 Rat and Mouse Ration ................................ 273 

APPENDIX L Sentinel Animal Program ..................................... 277 
4 Scopolamine Hydrobromide Trihydrate, NTP TR 445 
5 

ABSTRACT 

SCOPOLAMINE HYDROBROMIDE TRIHYDRATE 

CASNumber: 6533-68-2 

ChemicalFormula: C,,Hz2BrN0,*3H20 MolecularWeight: 438.33 

Synonyms: Scopolamine hydrobromide, 6,7-epoxytropan-3-yl, euscopol, hydroscine hydrobromide, hyoscine bromide, (-)-hyoscine 

hydrobromide, hysco, isoscopil, scopolammoniurn bromide, @)-tropate hydrobromide trihydrate, I-tropyl-cy-scopine 

Trade names: Beldavrin. Kwells, Sereen, Scopos, Triptone 

Scopolamine hydrobromide trihydrate is used in 

ophthalmic preparations and as a preanesthetic 

sedative. Its major use is in transdermal patches for 

the treatment of motionsickness.Scopolamine 

hydrobromide trihydrate wasselected for study 

becauseof considerable human exposure resulting 

from its use in prescription and over-the-counter 

preparations. Scopolamine was a suspect carcinogen 

because it contains an aliphatic epoxide moiety which 

mayact as a biological alkylatingagent.Maleand 

female F344/N rats andB6C3F,micereceived 

scopolamine hydrobromide trihydrate (89% pure) in 

distilled water by gavage for 16 days, 14 weeks, or 

2 years. Genetictoxicologystudieswereconducted 

in Salmonella typhimun'um,cultured Chinese hamster 

ovary cells, and mouse peripheral blood erythrocytes. 

16-DAYSTUDY IN RATS 

Groups of five male and five female ratswere 

administered 0, 75, 150, 300, 600, or 1,200 mg 

scopolamine hydrobromide trihydrate/kg body weight 

in distilled waterby gavage for 16 days.Allrats 

survived to the end ofthe study. The final mean 

body weights and body weight gains of males receiving 600 and 1,200 mg/kg and the mean body weight 

gain of males receiving 300 mg/kg were significantly 

lowerthanthoseof the control group. Clinical 

findingsincludedbilateral pupillary dilation in all 

dosed animals Ad red eyelids in males and females 

receiving 1,200 mg/kg. There were no significant 

treatment-related gross or microscopic lesions. 

16-DAYSTUDY IN MICE 

Groupsof five male and five female mice were 

administered 0, 150,250, 450, 900,or 1,800 mg 

scopolamine hydrobromide trihydrate/kg body weight 

in distilled water by gavage for 16 days. Onemale 

andtwofemalesreceiving 1,800 mg/kg and one 

femalereceiving 150 mg/kg died during the study. 

The final mean body weights and body weight gains 

of dosed micewere similar to those of the control 

groups. Clinical findings related to scopolamine 

hydrobromide trihydrate administration included 

bilateral pupillary dilation and squinting in all dosed 
6 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

malesandfemales.Therelativeliverweightsoflowing.Therewerenoothersignificantreatmentmalesreceiving 1,800 mg/kg and of femalesinallrelatedgrossormicroscopicfindings. 

dosed groups were significantly greater than those of 

thecontrol groups. Therewerenosignificant 

treatment-related gross or microscopic lesions. 14-WEEK STUDY IN MICE 

Groups of 10 male and 10 female mice were administered-0, 15, 45, 135, 400,or 1,200 mg scopolamine 14-WEEK STUDY IN RATS hydrobromide trihydratelkg body weight in distilled Groups of 10 male and 10 female rats were adminis- waterbygavage for 14 weeks.Onemalereceiving tered 0, 15, 45, 135, 400, or 1,200 mg scopolamine 135 mg/kg and two males and one female receiving hydrobromide trihydrate/kg body weight in distilled 1,200 mg/kg died during the study. The final mean water by gavage for 14 weeks. One female receiving bodyweightsandmeanbodyweightgainsofall 45 mg/kg, onemaleandonefemalereceiving dosed male groups and females receiving 45 mg/kg 135 mg/kg, six malesandonefemalereceiving and above were significantly lower than thoseof the 

400 mg/kg,andeightmalesandsevenfemales control Clinical included groups. observations receiving 1,200 mg/kg diedduringthestudy.The bilateral pupillary dilation, hyperactivity, and hypo- final mean body weights and mean body weight gainsactivity. ofalldosedmalesandfemalesweresignificantly 

lowerthanthoseofthecontrolgroups.Clinical A minimaltomildmatureneutrophilia,similarto findingsincludedbilateralpupillarydilationinall thatwhich occurredinthe14-weekratstudy, dosed males and females and reddening of the eyes inoccurred in male mice receiving45 mg/kg or greater. ' 15mg/kgmalesand135, 400, and1,200mg/kg As intheratstudy,therewasnomicroscopicevi- males and females. dence of inflammation thatcouldaccount for the 

neutrophilia. Hematocrit, concentration, hemoglobin and/or 

erythrocyte count in male and female rats receiving The estrous cycle lengthof 1,200 mg/kg females was 

45 mg/kg or greater were slightly higher than those significantly greater than that in the control group. 

of the controlgroups.Ingeneral,thesechanges 

weremostprominentinratsinthe 400 and There were no significant treatment-related gross or 

1,200 mg/kg groups. Higher hematocrit, hemoglobin microscopic lesions. 

concentration, and erythrocyte count were likely due 

tohemoconcentrationfromdehydration(relative 

erythrocytosis). A minim& to mild matureneutro- %YEAR STUDY IN RATS philia,evidenced by highersegmentedneutrophil Groups of 60male and 60 female rats were adminis- numbersthaninthecontrolgroup,occurredinall tered 0, 1, 5, or 25 mg scopolamine hydrobromide dosed male rats. trihydrate/kgbodyweight in distilled water by 

gavage for 104 weeks.Tenmalesandtenfemales Sperm morphology and vaginal cytology parameters from each dose group, excluding the1 mg/kg female indosedratsweresimilartothoseinthecontrol group, were evaluated at 15 months. groups. 

Ninemaleandfivefemaledosedratsdiedfrom Survival, Body Weights, Clinical Findings, 

esophagealobstructions consisting ofand feed and Ophthalmic Examination Findings 

bedding material in the posterior pharynx. Tracheal Thesurvivalratesoffemaleratsreceiving 1 and 

obstructionoccurredconcurrently withesophageal 25mg/kgwere significantlylower than that ofthe 

obstruction as a result of food build-up in the oro- control group. Mean body weights of 1 and 5 mg/kg 

pharyngealregion.Thiscondition is considered to malesandfemalesweresimilar to those of the 

be secondary to the inhibitory effects of scopolamine controls throughout the study. However, mean body 

hydrobromide trihydrate on salivary gland secretions weights of 25mg/kgmalesandfemalesweregenand on esophageal smooth muscle involved in swal- erallylowerthanthoseofthecontrolgroupsafter 
Scopolamine Hydrobromide Trihydrate, NTP TR 445 7 

about week 25. Clinicalfindingsincludedbilateral 21/50,26/50,24/50; females: 20/50,6/60, 13/50, 

pupillary dilation in all dosedmalesandfemales. 4/50). Theincidenceofmononuclearcellleukemia 

Ophthalmicexaminationrevealednosignificantinfemalesreceiving 25 mg/kg was wellbelow the 

findings. NTP historical range. 

Hematology 

Compared to controls, hematocrit was slightly higher %YEAR STUDY IN MICE 

inthe 25 mg/kg male rats, similar to the effects Groups of70 male and70 female 'mice were adminisobserved in the 14-week study; this is consistent with tered 0, 1, 5, or 25 mg scopolamine hydrobromide dehydration resulting inhemoconcentration.Retic- trihydrate/kg body weightindistilledwaterby ulocyte numbers in the 25 mg/kgfemale rats were gavage for 104 to 105 weeks. Ten control animals slightly lower than those in the controls. This result and ten animals from each dose level were evaluated 

is consistent with the lower body weights, and thus a at 15 months.

decreased nutritional status,exhibitedbythese 

animals. Survival, Body Weights, Clinical Findings, 

and Ophthalmic Examination Findings Plasma Scopolamine Determinations Survival of dosed males and females was similar to The serum scopolamine concentrationswere 6 ng that of the controls. The mean bodyweightsof scopolamine/mL serum for the 5 mg/kgfemale males and females receiving 1 mg/kg were similar to sample and 12 and 28 ng/mL for the 25 mg/kg male those of the control groups throughout the majority andfemale samples, respectively. Theamounts of ofthe study. The mean bodyweightsof 5 mg/kg scopolamine in the other serum samples were below males and females were slightly lower than those of . the minimum detection limit (4 ng/mL) of the analy- the controls. Themeanbodyweightsofmalesand sis method. females receiving25 mg/kg were lower than those of 

the control groups after week 13. Clinical findings Neurobehavioral Findings included bilateral pupillary dilation in all dosed male Horizontal motor activity of 25 mg/kg females was and female groups. Ophthalmic examination revealed significantly greater than that of the control group on no significant findings. days 90,180, and 360. Startle response of 5 and 

25 mg/kg females was significantly lower than that of 

the control group on day 90. On day 180, passive Hematology 

avoidance of25 mg/kg males was significantly lower Hematocrit, hemoglobin concentration, and erythrothan that of the control group. cytecountin 25 mg/kgfemalemice were slightly 

lower than those in the control group. These results 

are consistent with development ofa minimal normo- Pathology Findings cytic, normochromic nonresponsiveanemia. The The incidences of adenoma of the pituitary gland pars anemia may be related to the lower body weights distalis decreased with increasing dose in both male 

exhibitedbytheseanimalsand are presumed to be and female'rats; however, this trend was only signifi- due to a decreased nutritional status. cant in males vehicle (males: control, 19/49;

1 mg/kg, 17/49; 5 mg/kg, 13/50;25 mg/kg, 10/50; 

females: 20/50,13/60,14/50,10/50). Theinci- Pathology Findings 

dences of adenoma of the pituitary gland pars distalis The combined incidences of hepatocellular neoplasms 

in 25 mg/kg males and all groups of dosed females (adenoma or carcinoma) occurred with a significant 

werebelow the NTP historical control range. The negative trend in males and females (males: vehicle 

incidencesofhyperplasiawerenotsignificantly control, 30/50; 1 mg/kg, 33/50; 5 mg/kg, 14/50; 

different from those in the control groups. 25 mg/kg, 15/50; females: 22/51,21/50,16/50, 

9/51). The combined incidencesof hepatocellular 

The incidences ofmononuclearcelleukemiain neoplasms in 5 and 25 mg/kg males were within the 

25 mg/kg males and females were significantly lower NTPhistorical control range. The incidences of 

thanthoseofthe control groups (males: 33/50, clear cell foci and eosinophilic foci in dosed male 
8 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

groups,and eosinophilicfociin 25 mg/kg females, 

weresignificantlylowerthanthose ofthecontrol 

groups. 

Theincidences ofmanyspontaneouslyoccurring 

nonneoplasticlesionsweresignificantlylowerin 

dosedmicethaninthecontrolgroupsandusually 

decreasedwithincreasingdose.Theseincluded 

kidney nephropathy, alveolar epithelial hyperplasia, 

hyperplasiaofthepancreaticislets,bonemarrow 

myelofibrosis, hyperplasia of the pituitary gland pars 

distalis, cystic hyperplasia of the uterus, and hematopoieticcellproliferation of the spleen.Thedecreased incidences of these spontaneous lesions were 

most likely a result of lower body weights in dosed 

animals. 

GENETIC TOXICOLOGY 

Scopolamine hydrobromide trihydrate didnot induce 

mutations in any of five strains of Salmonella typhimurium, with or without S9 metabolicactivation 

enzymes,inducedi sisterchromatid norit 

exchangesinculturedChinesehamsterovarycells, 

with or without S9. A weakly positive response was 

obtained, however, in a chromosomal aberrations test 

conductedinculturedChinesehamsterovarycells 

with very high doses of scopolamine hydrobromide 

trihydrateinthepresence of S9; without S9, no 

increaseinaberrationswasnoted.Despitethe 

evidence for chromosomal damage observedin vitro, 

noincreaseinthefrequenciesofmicronucleated 

normochromatic erythrocytes was observed in peripheral blood samples of male or female mice exposed 

to scopolamine hydrobromide trihydratefor 14 weeks 

by gavage. 

CONCLUSIONS 

Under the conditions of these 2-year gavage studies, 

there was noevidence of carcinogenicactivity* of 

scopolaminehydrobromidetrihydrateinmale or 

female F344/N rats or B6C3F, mice administered 1, 

5, or 25 mg/kg. 

* Explanation of Levels of Evidence of Carcinogenic Activity is on page 10. A summary of the Technical Reports Review Subcornminee 

comments and the public discussionon this Technical.Report appearson page 12. 
Scopolamine Hydrobromide Trihydrate, NTP TR 445 9 

Summary of the 2-Year Carcinogenesis and Genetic Toxicology Studies 

of Scopolamine Hydrobromide Trihydrate 

Male

F344m Rats 

Female 

F344/NRats 

Male 

B6C3Fi Mice 

Female 

B6C3Fi Mice 

Doses 0, 1. 5, or 25 mglkg 

in water by gavage 

0,1, 5, or 25 mglkg 

in water by gavage 

0,1, 5, or 25 mglkg 

in water by gavage 

0, 1. 5, or 25 mglkg 

in water by gavage 

Bodyweights 25 mglkggrouplower 25 mglkggrouplower 5 and 25 mglkg 5 and 25 mglkg 

thancontrolgroupthancontrolgroupgroupslowerthan groups lower than 

control group control group 

2-Year survival 20/50.14/50, 22/50. 34/50, 17/60. 26/50.40150, 39/50, 39/50, 33/51, 36/50. 37/50, 

rates 28/50 22/50 39/50 38151 

effects 

Nonneoplastic None None None None 

Neoplasticeffects None None None None 

Levels of evidence Noevidence No evidence No evidence No evidence 

of carcinogenic 

activity 

Genetic toxicology 

Salmonella typhimuriurn gene mutations: Negative with and without S9 in strains TA97, 

TA1537 

TA98, TAW,TA1535,and 

Sister chromatid exchanges 

CulturedChinesehamsterovarycells in vitro: Negativewithandwithout S9 

Chromosomal aberrations 

CulturedChinesehamsterovarycells in vitro: Negativewithout S9;weaklypositivewith S9 

Micronucleated erythrocytes 

Mouseperipheral,blood in vivo: Negativeinmaleandfemalemice 
10 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

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: b o 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. 

0 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. 0 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. 0 Equivocal evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing a marginal increase 

of neoplasms that may be chemical related. 0 No evidence of carcinogenic activity is demonstrated by studies that are interpreted as showing no chemical-related 

increases in malignant or benign neoplasms. 0 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 

betweentwo adjacent levels. Theseconsiderations shouldinclude: 

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 knownor 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 neoplasiaor 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 positiveor false negative concerns; 

structure-activity correlations; and 

in some cases, genetic toxicology. 
Scopolamine Hydrobromide Trihydrate,NTP TR 445 11 

NATIONAL TOXICOLOGY PROGRAM BOARD OF SCIENTIFIC COUNSELORS 

TECHNICAL REPORTS REVEW SUBCOMMITTEE 

The members of the Technical Reports Review Subcommittee who evaluated the drafi NTP Technical Report on scopolamine hydrobromide 

trihydrate on June 20, 1995, are listed below. Subcommittee members serve as independent scientists, not as representatives of any 

institution, company, or governmental agency. In this capacity, subcommittee members have five major responsibilities in reviewing NTP 

studies: 

l 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, 

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

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

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

Arnold L.Brown, M.D., Chairperson 

university of Wisconsin Medical School 

Madison, WI 

Thomas L. Goldsworthy, Ph.D. 

Department of Experimental Pathology and Toxicology 

Chemical Industry Institute of Toxicology 

Research Triangle Park, NC 

Meryl H.Karol, Ph.D. 

Department of Environmental Occupational Health 

University of Pittsburgh 

Pittsburgh, PA 

Curtis D. Klaassen, Ph.D., PrincipalReviewer 

Department of Pharmacology and Toxicology 

University of Kansas Medical Center 

Kansas City, KS 

Claudia S. Miller, M.D., M.S. 

University of Texas Health Sciences Center 

San Antonio, TX 

Janardan K.Reddy, M.D. 

Department of Pathology 

Northwestern University Medical School 

Chicago, IL 

Irma RUSSO, M .D .,Principal Reviewer 

Fox Chase Cancer Center 

Philadelphia, PA 

Louise Ryan, Ph.D. 

Division of Biostatistics 

Harvard School of Public Health and 

Dana-Farber Cancer Institute 

Boston, MA 

Robert E. Taylor, M.D., Ph.D.. PrincipalReviewer 

Department of Pharmacology 

Howard University College of Medicine 

Washington, DC 

Mary Jo Vodicnik, Ph.D. 

University of Indiana 

Indianapolis, IN 

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

National Cancer Institute 

Frederick, MD 
12 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

SUMMARY OF TECHNICAL REPORTS REVIEW SUBCOMMITTEE COMMENTS 

On June 20, 1995, the Technical Report on the 

toxicology and carcinogenesis studies of scopolamine 

hydrobromide trihydrate received public review by 

theNationalToxicology Program’s Boardof 

Scientific Counselors’ Technical ReportsReview 

Subcommittee. The review meetingwasheld at the 

National Institute of Environmental Health Sciences, 

Research Triangle Park, NC. 

Dr. K.M. Abdo, MEHS, introduced the toxicology 

and carcinogenesis studies of scopolamine hydrobromide trihydrate by discussingtheusesofthe 

chemicaland rationale for study, describing the 

experimental design, and reporting on survival and 

bodyweight effects. The proposed conclusions for 

the two-year gavage studies were no evidence of 

carcinogenicactivity ofscopolamine hydrobromide 

trihydrate in male or female F344/N rats or B6C3F, 

mice administered 1, 5, or 25 mg/kg. 

During the 16week study, rats receiving the higher 

doses experienced considerable mortality attributed to 

esophageal and tracheal obstruction by feedand 

bedding. This condition was considered secondary to 

the inhibitory effects of scopolamine on salivary 

secretion and the motility of smooth muscle involved 

in swallowing. 

Dr. Klaassen, a principal reviewer, agreed withthe 

proposed conclusions. Hewas surprised that more 

scopolamine could be administered by gavage than in 

thefeed and asked for elaboration. Dr. Abdo explained that the problem with esophageal obstruction, 

even at lower doses, diminished theamountthat 

could be administered inthe feed. Dr. Klaassen 

noted that a decrease in body weight is often given as 

a explanation for a decrease in neoplasmincidence 

such as reported here, and suggested creating a graph 

thatwould let the reader evaluate thisconclusion. 

Dr. J.K. Haseman, NIEHS, agreed this was a good 

ideaandsaidhewouldevaluatetwopotentialapproaches to illustrate this association, one using a 

modelbased , on logistic regression to predict 

neoplasm incidence for animals at a particular body 

weight, and the other looking empirically at a moving 

averageofneoplasmincidence for animals inthe 

database of a given weight range (Table 16). 

Drs. Russo and Taylor, the other principal reviewers, 

agreedwith the proposed conclusions. They were 

pleasedthatthe report included neurological data 

allowing evaluation of neurobehavioral toxicity and 

data from pharmacokinetic studies enabling the 

correlation of toxic effects with plasma levels. 

Dr. Taylor questioned listingall therapeutic uses 

associated with scopolamine as in his experience the 

drug had notbeenemployed for a number of the 

listed conditions during the last 20 years. Dr. Abdo 

said he would use the pasttense to describe therapeutic uses that no longer apply. 

Dr. R. Hart, NCTR, observed that body weight 

reductionsmayhave a sparing effect relative to 

neurobehavior and other neurological endpoints. 

Dr. Goldsworthy strongly supported inclusion of 

pharmacokinetic studies, particularly encouraging 

their use in a more prospective fashion to help set 

dose levels for better comparisons across species. 

Dr. G. Lucier, NIEHS, agreedandsaid that an 

interdisciplinary toxicokinetics facultyhas been established within the NIEHS to help determine the kinds 

ofspecificstudiesthatshould be incorporated into 

the study design for a chemical. In some cases the 

dataobtainedwouldaidin the development of a 

biologicallybased dose-response model. Dr. Miller 

said it would be useful to include actual drug doses 

and average duration of treatment commonly used in 

medical practice. 

Dr. Klaassen movedthat the Technical Report on 

scopolamine hydrobromide trihydrate be accepted 

with the revisions discussed and with the conclusions 

as written for male andfemale rats and mice, no 

evidence of Carcinogenic activity. Dr. Taylor seconded the motion, which was accepted unanimously 

with ten votes. 
13 

INTRODUCTION 

A H 

SCOPOLAMINEHYDROBROMIDE TRIHYDRATE 

CAS Number: 6533-68-2 

ChemicalFormula: C,,H,,BrN04*3H,0 Molecular Weight: 438.33 

Synonyms: Scopolamine hydrobromide, 6.7-epoxytropan-3-yl. euscopol, hydroscine hydrobromide, hyoscine bromide, (-)-hyoscine 

hydrobromide, hysco, isoscopil, scopolammonium bromide, (s)-tropate hydrobromide trihydrate, I-tropyl-a-scopine 

Trade names: Beldavrin, Kwells, Sereen, Scopos, Triptone 

CHEMICAL AND PHYSICAL PROPERTIES 

Scopolamine hydrobromide trihydrate occurs as 

colorless or white crystals or as a white granular 

powder with a melting point of 195 O to 199" C. It 

is soluble in water (66.7 g/100 mL) andalcohol 

(5 g/100 mL), slightly soluble in chloroform, and 

almost insoluble in ether (Merck Index, 1989). 

PRODUCTION,USE, 

AND HUMANEXPOSURE 

Scopolamine (hyoscine) is a solanaceous alkaloid that 

is a natural product of plants of the Solanaceae 

family (Brown, 1990). It is the levorotatory form of 

the parent compound, atroscine. Scopolamineisan 

organic ester formed>bythecombination of an 

organic acid, tropic acid, and a complex organic 

base, scopine or 6,7-epoxytropine (Gearien, 1989). 

It is found chiefly in the shrubs Hyoscyamusniger 

(henbane) and Scopolia carniolica. Although scopolamine can be recovered from the mother liquors 

remaining after the crystallization of hyoscyamine, a 

tropic acid ester of tropine, it is routinely extracted 

from the leavesof Datum mete1 in India or from 

Daturameteloides inMexicoandimported to the 

United States(Remington's Pharmaceutical Sciences, 

1975); data on the amount importedare not available. 

The accidental consumptionof roots from Datura 

stramonium, which is commonly referred to by a 

varietyofnamesincluding apple of Peru, Jimsonweed,Jamestownweed, devil's apple, thorn apple, 

stinkweed, stramonium, and loco weed, has produced 

acutetoxicepisodes(Labiancaand Reeves, 1984; 

Brown, 1990). 

Though no figures on the production of scopolamine 

or its derivatives were reported to the U.S. International Trade Commission from 1974 through 1977, 

the United States production during this interval was 

estimated to be less than 4.5 x le grams (USITC, 

1977). The estimated overall oral and dermal human 

exposure to scopolamine and its salts (scopolamine 

hydrobromide, methyl scopolamine bromide, and 

methyl scopolamine nitrate) was 1.6 x los grams per 

year (NCI/SRI, 1979). A national prescription audit 

estimated that during 1980, 14,000 prescriptions were 
14 

writtenhe States United in for ophthalmic

preparationscontainingscopolamine(NDTI, 1980). 

Approximately5.9 million prescriptions were written 

in 1986 for all scopolamine formulations (Dr. Vera 

Glocklin,FoodandDrugAdministration,personal 

communication to K.M. Abdo).Morerecentdata 

were not available. 

Scopolamine is used primarily for its antimuscarinic 

drug properties in the peripheral and central nervous 

system. Its major mechanismofactionisvia a 

competitiveantagonismwiththeneurotransmitter 

acetylcholineateffectorsites(exocrineglandsand 

smooth and cardiac muscle) (Brown, 1990). 

Central nervous system effects resulting from theraScopolamine Hydrobromide Trihydrate, NTP TR 445 

pharmacologically associated with its abilityto inhibit 

gastrointestinal and respiratory tract mucus secretions 

and its antispasmodic ability to relax smooth muscle 

in the respiratory tract, urinary bladder, and gastrointestinaltractincludingthegallbladder(Brown,' 

1990). Scopolaminehasbeenusedincoldand 

allergy medications and asa preanesthesia medication 

for its ability to inhibit mucous membrane secretions 

oftheoraland respiratorypassages(PDR, 1992). 

Scopolamine'santispasmodiceffects are usedin 

combinationwithantibioticsandanalgesics for the 

treatment of urinary tract infections. 

Becausescopolamineinhibitsgastricsecretionsand 

motility, it is particularly useful in treating ulcers and 

functionaldiarrhea.However,thetherapeuticdose 

peutic doses of scopolamine characteristically includefor such treatmentisoftenhighandresultsin 

drowsiness, euphoria, amnesia, fatigue, and a reduc- undesirableperipheral and' central nervoussystem 

tioninrapid-eye-movementsleep(NRC, 1982; side effects (Brown, 1990). Quaternaryammonium 

Brown, 1990). Excitatoryeffectsregularlyoccur derivatives of scopolamine(methyl ' scopolamine 

afterlargenontherapeuticdosesand,in a small bromideandmethyl nitrate) scopolamine are 

number of cases, have occurred unexpectedly follow-indicated for such treatment to eliminate side effects 

ingtherapeuticdoses (NRC,1982). Theseeffects 

includeexcitement,restlessness,hallucinations,and 

delirium. 

Scopolamineuseassociatedwithitssedativeand 

othercentralnervoussystemeffectsincludesthe 

treatment of acute mania sind delirium; the symptomatictreatment of infantile cerebralpalsy;thetreatment of paralysis agitans and spastic states resulting 

from nervous system injuries; and in the diagnosis of 

psychomotor epilepsy (Wade,1977;Reynolds, 1982). 

Scopolamine was previously included in many overthe-counter sleep aids, but was later withdrawn due 

toinadequatefficacyratingscoupledwiththe 

potential for adverse anticholinergic effects. Scopolaminehasbeenused for the treatment of postencephaliticparkinsoniantremorsandbyobstetriciansduringdeliverycombination in with 

morphine to induce a state of. amnesia andpartial 

analgesiareferred to as ."twilightsleep";however, 

scopolamine is no longer the drug of choice in these 

cases.Themajoruse of scopolamine is for the 

prevention of vestibular disorderssuchasmotion 

sickness when given transdermally. 

Peripheral nervous system effects that contribute to 

thetherapeuticpotentialscopolamine of are 

involving the central nervous system; however, such 

derivatives usually require parenteral administration 

to achieve acceptable results. 

Althoughscopolaminehydrobromidehasnotbeen 

officiallyapprovedbythe.FDA for therapeutic 

ophthalmic use, this use for scopolamine was under 

investigation(personalcommunicationfromRobert 

Linkous,FDA, 1989). Scopolamineislisted as an 

ophthalmic drug to be used exclusivelyfor diagnostic 

purposes. blockscholinergic Scopolamine the 

response of the sphincter muscle of the iris and the 

muscleoftheciliarybody,resulting in pupillary 

dilation (mydriasis) and paralysis of accommodation 

(cycloplegia) (Wade, 1977; Reynolds, 1982; Brown, 

1990). Indications for use cycloplegic include

refractionandpupillarydilation for postoperative 

inflammatoryconditionsoftheirisanduvealtract 

and to break synechia.Due to its longduration of 

action for 7 to 12 days (Brown, 1990), ' scopolamine 

hydrobromideisnotrecognizedtobethedrug of 

choice for ophthalmic use. 

Scopolaminehydrobromidehasoccasionallybeen 

usedinveterinarymedicineincombinationwith 

morphine as preanesthetic in a sedativedogs

(Remington's Pharmaceutical Sciences, 1975). 
Introduction 15 

PHARMACOKINETICS,ABSORPTION, 

DISTRIBUTION, METABOLISM, 

AND EXCRETION 

Structure-Activity Relationship 

There are no structural relationships between scopolamine or its salts and anyknowncarcinogens. 

Scopolamine has been suggested as a possible carcinogen because it contains an aliphatic epoxide moiety, but there is no evidence for its alkylating activity 

under physiological conditions (Ehrenberg and 

Hussain, 1981; Connors, 1984). The structureactivity relationships relative to the general class of 

anticholinergic drugs and their binding to muscarinic 

receptors have beenextensively described, and are 

discussed below (Abramson et al., 1969; Cullumbine, 

197 1; Abramsonet al., 1974; Beld and Ariens, 1974; 

Lien et al., 1976; Gearien, 1989; Brown,1990). 

There are several critical structural components of 

the scopolamine molecule thatre involved in absorption, distribution, and pharmacological activity. The 

intact ester of tropic acid withthe organic base, 

scopine, is essential for scopolamine's antimuscarinic 

activity. The presence of a free hydroxyl group in 

theacid portion of the ester is also important for 

enhancedbiologicalactivity.Demethylation of the 

nitrogen moiety reducesactivity,whilemethyl 

quaternizationincreasesbiologicalactivity.Quaternaryammonium derivatives are morepotentin 

general than the parent compound upon parenteral 

administration but are poorly and unreliably absorbed 

orally and lack central nervous system activity due to 

poor penetration into the brain. Quaternizationby 

higher alkyl groups diminishes biologicalactivity. 

The asymmetrical carbon atom immediately adjacent 

to the benzene ring portion of tropic acid is important 

relative to the biological activity of scopolamine; i.e., 

scopolamine is L-hyoscine which is much more active 

thanD-hyoscine. The acyl group or tropic acid 

portion of the molecule is considered responsible for 

the blocking action at the muscarinic receptors. 

Experimental Animals 

Studies have beenperformed that characterize scopolamine metabolism in the mouse, rat, guinea pig, and 

commonmarmoset (Werner and Schmidt,1968). 

When [9-14C]- and [9-3H,]-scopolamine hydrobromide 

were administered by an intraperitoneal injection to 

mice,radioactive scopolamine-9,l-/.?-D-glucuronide, 

aposcopolamine, 6-hydroxyhyoscyamine, scopine and 

unchanged scopolamine, and nonradioactive norscopolamine and norscopolamine-9,1-0-D-glucuronide

were recovered from the urine. After intraperitoneal 

injections of [9-'4C]-scopolaminehydrobromide, 23 % 

of theradioactivityoftheN-methyl group was 

expired as 14C02 by themouse, 5.9% by the rat, 

12.5% bytheguinea pig, and 4.5 % bythemarmoset. There was no racemization of scopolamine or 

anyof its optically active metabolites. The scopine, 

6-hydroxyhyoscyamine,and scopolamine-9,l-p-Dglucuronide fraction of mouse urine contained more 

radioactivity 25 hours after the oral administration of 

scopolamine hydrobromide than after the intraperitonealrouteof administration. Overall, the 

majormetaboliteofscopolaminewasshown to be 

scopolamine-9,1-p-D-glucuronide(Bayne et al., 

1975). An enzymecapable of hydrolyzing scopolamine, referred to as a tropinesterase for its major 

action, has been detected in various tissues including 

theserum of rabbitsandthe liver of guinea pigs 

(Cullumbine, 1971). In rabbits theabilityof this 

enzyme to hydrolyze atropine is inherited through a 

genethat is incompletely dominant, and it is associated with another gene that influences the amount 

of black pigmentin the fur. Scopolamine-N-butyl 

bromide undergoes biliary excretion in rats (Klaassen 

et al., 1981); 21% of an intravenous dose (6 mg/kg) 

was excreted by this route within 4 hours. 

Humans 

In humans, scopolamine hydrobromide is readily 

absorbed from the gastrointestinal tract and enters the 

circulationwhenapplied locally to mucosal 

membranes(Wade, 1977; NRC, 1982; Reynolds, 

1982;Brown, 1990). Limited absorption occurs 

from the intact skin and the eye. Once in the systemic circulation, it readily crosses the blood-brain 

barrier (MA, 1977). It binds to plasma proteins 

and is almost entirely metabolized in the liver (Wade, 

1977;Reynolds,1982).Peak serum concentrations 

are observed within 1 to 2 hours after an oral or 

intramuscular dose of scopolamine hydrobromide 

(Brand,1969;Bayne et al., 1975;Wade, 1977). 

Onlyabout 1% of an oral dose of scopolamine is 

eliminated as the parent compound in the urine 

(NRC, 1982). 

The pharmacokinetics and bioavailability of scopolamine hydrobromide were investigated in six healthy 
16 

male subjects receiving oral or intravenous doses of 

0.4 mg (Putcha et al., 1989). Afterintravenous 

administration, plasma concentrations of scopolamine 

declinedin a biexponential fashion, with a rapid 

distribution phase and a comparatively slow elimination phase. Mean values for volume of distribution, 

systemic clearance, andclearance renal were

1.4 Wkg, 65.5 Whour, and 4.2 L/hour, respectively. 

Meanpeakplasma concentrations were 2.9 pg/mL 

following intravenous doses and 0.5 pg/mL following 

oral doses. The elimination half-life was 4.5 hours. 

Bioavailability of the oral doses was variable among 

subjects, ranging between 10.7% and 48.2%. 

Ophthalmic effects due to local action of the scopolamine may persist for 3 to 7 days whereas systemic 

effects from oral or parenteral doses last 4 to 6 hours 

(Cullumbine, 1971). Eventhoughthe literature 

refers to an almost complete metabolism of scopolamine by humans, there are no studies documenting 

themetabolicpathways or characterizing the major 

and minor metabolites. It is known that scopolamine 

hydrobromide administered during the:first stage of 

labor crosses the placental barrier, and traceshave 

been detected in the milk of exposed nursing mothers 

(Wade, 1977; Reynolds, 1982). 

TOXICITY 

Experimental Animals 

The LD,, values for scopolamine hydrobromide listed 

in Table 1 have been determined with various routes 

of administration in different species (RTECS,1982). 

Scopolamine Hydrobromide Trihydrate,NTP TR 445 

Humans 

The 1984 edition of Clinical Toxicologyof Commercia1 Products (Gosselin et al, 1984) assigned a 

toxicityratingof five (extremely toxic) to scopolamine; previous editions (Gleason et al., 1969; 

Gosselin et al., 1976) assigned the higher toxicity 

rating of six (supertoxic). Both ratings underline the 

acutetoxicnatureof scopolamine. Although fatalities from exposure are rare (Gosselin et al., 1984), 

they have occasionally occurred, usually in children 

(Remington's Pharmaceutical Sciences, 1975; NRC, 

1982). A fatal human dose is estimated to be about 

8 mg/kg, withdeathusually occurring within 

24 hours of the initial exposure (Theines and Haley, 

1972). Compared to LD,, valuesnoted in experimental animals, humans appear more susceptible to 

thelethalactionof scopolamine than experimental 

animals.Symptomsoftoxicity in humans occur 

promptly and may persist for hours or days. Very 

low doses of scopolamine elicit a variety of adverse 

acute including reactions, idiosyncratic effects, 

followingtherapeutic doses. Accidental poisonings 

.from as little as four drops of 0.25% -scopolamine 

solution, equivalent to 0.45 mg ,scopolamine 

(Goldfrank et al., 1982), from a single 0.5 mg 

transdermalpatchof scopolamine (Rodysill and 

Warren, 1983), and from a 0.3 mgKwells motion 

sicknesstablet (Hindson, 1958) havecaused acute 

toxic psychosis. 

Scopolamine exposure can produce symptoms of 

acuteoxicityinvolving the peripheral nervous 

system, including thirst due to drying of the mouth 

TABLE1 

Scopolamine Hydrobromide Toxicity&Dm)in Experimental Animalsa 

Species 

Rat 

Rat 

Rat 

Mouse 

Mouse 

Mouse 

Mouse 

Guinea Pig 

a RTECS, 1982 

Route Dose (mgkg) 

Oral 1,270 

Subcutaneous 3,800 

Intraduodenal 670 

Oral 1,880 

Intraperitoneal 650 

Subcutaneous 1.650 

Intravenous 203 

Subcutaneous 850 
Introduction 17 

and throat, photophobia caused by dilatedpupils, loss 

of accommodation for near vision (blurred vision), 

increased intraocular pressure (glaucoma), fever due 

to flushing and drying of the skin, urinary retention, 

and bradycardia (25-200 mg) or tachycardia followed 

by bradycardia (300-600 mg) (ThienesandHaley, 

1972; Wade, 1977; Reynolds, 1982; Brown, 1990). 

Central nervous systemmanifestations of acute 

scopolamine exposure include both cerebral excitementand depression. After large doses, the excitement component is less commonly noted and usually 

occurs earlier and is of shorter duration than that 

observed after smaller doses. This component may 

be expressed as euphoria, delirium, hyperactivity, 

disorientation, psychotic behavior, and hallucinations. 

The depressive cerebral component following therapeutic doses is expressed by an amnesic and sedative 

hypnoticstatewhile toxic doses may elicit depression, muscular weakness, disequilibrium, and coma. 

Chronic toxic symptoms include recent memoryloss, 

mental confusion, and hallucinationswithrepeated 

episodes of theta wave activity in the electroencephalogram (Thienes and Haley,1972).Psychological 

dependence may occur but discontinuance of drug 

exposure does not induce physical withdrawal symptoms (Goldfrank et al. , 1982). A 1-weekrecovery 

periodisrequired to remove symptoms of chronic 

exposure (Thienes and Haley, 1972). 

Rare cases of scopolamine-induced hypersensitivity 

have been documented following mucous membrane 

exposures to eye drops; this hypersensitivity was 

' characterizedbyedemaofthe uvula, glottis, and lips 

aswell as generalized urticaria (Guill et al., 1979). 

Sensitized patients become more sensitiveto the drug 

after repeated exposures, as manifested by

ever-decreasing exposure times required to elicit the 

allergic response. 

During the years 1969 to 1980, a total of 37 adverse 

reactions to scopolaminewere documented 

(FDA 1988). Twenty-three of these reports indicated 

nervoussystem disorders and 14 outlined cardiovascular disorders. Toxicpsychosisoccurredin a 

67-year old femalewithinsulin-dependentdiabetes 

whoreceived six to eight drops of a cycloplegic 

ophthalmic solution consisting of atropine 

(20 mg/mL), scopolamine hydrobromide (5 mg/mL), 

symptoms abated after intravenous administration of 

physostigmine (Kortabarria et al., 1990). Toxic 

comaoccurredin a 6-year old boy who was given 

two drops per eye of an ophthalmic solution containing 2% atropine, 0.5 % scopolamine hydrobromide, 

and 45% phenylephrine hydrochloride. The patient 

recoveredfollowingtreatmentwithphysostigmine 

salicylate(two 9.5 mg intravenous injections given 

10 minutes apart; Nadal et al., 1987). 

Fetotoxicity can result from scopolamine exposure. 

In one study, scopolamine (0.43 mg) wasadministered alone or in combination with sedatives, analgesics, or both to 56 pregnant women during labor and 

the fetal heart rate was evaluated witha fetal monitor 

(Boehm and Growdon, 1974). There was a decrease 

in fetal heart rate baseline variability (an indication of 

fetal distress) in 42% of the patients treatedexclusively with scopolamine and in 75 % of those patients 

receivingscopolamineincombinationwith other 

drugs. Scopolamine toxicitywas also reported in a 

newbornwhosemotherreceivedmultiple doses of 

scopolamine(twodoseseach of 0.2, 0.3, and 

0.4 mg)with merperidine, lorfan, and "general 

inhalation"(Evensand Leopold, 1980). The toxic 

symptomsincluded an increased body temperature 

(100.4"F) and pulse rate (200 beatdminute),

lethargy, and a barrel-chested appearance without 

respiratory distress. 

REPRODUCTIVE 

AND DEVELOPMENTALTOXICITY 

Experimental Animals 

Thepotentialadverseconsequencesofscopolamine 

exposure on fetaldevelopmentand reproductive 

parametershavebeeninvestigatedinexperimental 

animal studies including those sponsored bythe NTP. 

In a study using CF-1 mice, scopolamine was found 

to be "non-teratogenic in the doses employed" 

(Iuliucci, 1973). However, when administered 

togetherwithmorphine sulfate, scopolamine enhancedmorphine-inducedexencephaly in the offspring. In a study conductedwithNewZealand 

White rabbits, pregnant does were given scopolamine 

hydrobromide in drinkingwater (0.010 mg/mL) or 

untreateddrinkingwater from the 10th through the 

andphenylephrine hydrochloride (40 mg/mL). The14thday of gestation (McBride et al., 1982). The 
18 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

calculated intake of scopolamine ranged from 0.424 thenumberof embryo deaths was also observed 

to 0.582mg/kg per day for the eight treated animals whenthe control group wascomparedwith the 

in the study. No malformed pups were observedin combined treatment groups. 

the control group. On gestation day 22,scopolamine 

significantly increased the incidence of malformationsThe NTP sponsored teratology studies of scopolamine 

in the treated group. In two ofthe eight treated 

litters, all of the live fetuses exhibited malformations. 

No malformed live fetuses were observed in theother 

six treated litters. Malformations observed inthese 

two litters includedexencephaly, hydrocephaly, 

buphthalmia, and microphthalmia. 

In a companion study (McBrideet al.,1982),scopolamine hydrobromide wasadministeredintramuscularly at 12 hour intervals to one or two pregnant 

NewZealand White rabbits at doses ranging from 

0.037 to 0.185mg/kg per injection on gestation 

days 10 through 14. The eight untreated animals in 

the drinking water study also served as controls for 

this study. The doe receiving 0.137mg/kg per 

injection died on the first day of treatment and the 

doe receiving 0.185mg/kg per injection died on the 

second day of treatment. Both does died of tachycardia induced by the injection. The other does exhibited clinical signs of distress, which the authors 

attributed to cardiac effects of the drug. On gestation 

day 22,no malformed fetuses were observed in any 

of the 38 live fetuses examined. The authors also 

noted a significant increase in the incidence of 

resorptions when the data from the combined treatment groups (oral and intramuscular) were compared 

to the control group. 

McBride et al. (1982)also described a study in which 

sterile saline or 0.1 or 0.2mg scopolamine hydrobromide was injected into white leghorn chicken eggs 

(100eggdgroup) after 96 hours of incubation. The 

chick embryos were then examined on day 12 of 

incubation. The incidences ofmalformationwere 

0%, 34%, and 78% for the control, 0.1 mg,and 

0.2mg treatment groups, respectively. The primary 

malformation in both scopolamine-treated groups was 

gastroschisis, with allofthemalformed embryos 

exhibiting this defect. Other malformations observed 

were reduction deformities of the leg, wing deformities, and microphthalmia. A significantincrease in 

hydrobromide using time-pregnant CD-1 mice and 

CD rats (data on file at NIEHS). Scopolamine 

hydrobromide (0, 10, 100, and 450, 900 mg/kg per 

day) dissolved in water was administered by gavage 

on gestation day 6 through gestation day 15. The 

animalswere sacrificed and cesarean sections were 

performed on gestation day 17 (mice) and gestation 

day 20 (rats). The results of these studies showed 

that rats exposed in ufero to 450 or 900 mg/kg 

scopolamine hydrobromide developed short ribs and 

that in utero exposure of rats to 100, 450,and 

900mg/kg scopolamine was associated with marginal, non-dose-related reductions in fetal body weight 

and marginal, non-dose-related increases in the 

incidences of fetal malformations in the presence of 

maternaltoxicity (reduced body weight and weight 

gain). The investigators concluded that there was no 

evidence of teratogenesis in mice, even though a high 

levelofcongenital malformations and anatomical 

variationswasnoted. Maternal and fetal body 

weightsofmicewerereducedat the 450 and 

900mg/kg dose levels. 

Humans 

No information on the reproductive or developmental 

toxicityofscopolamine hydrobromide trihydrate in 

humans was found in the literature (NLM, 1994). 

CARCINOGENICITY 

Experimental Animals 

No information on the carcinogenicity of scopolamine 

hydrobromide trihydrate in experimental animals was 

found in the literature (NLM, 1994). 

Humans 

No epidemiological studies or case reports examining 

therelationshipbetween exposure to scopolamine 

hydrobromide and human cancer were found in the 

literature (NLM , 1 994). 
Introduction 19 

GENETICTOXICITY 

The mutagenicity data for scopolamine hydrobromide 

trihydrate found in the literature were limited to two 

published reports. Fluck et al. (1976) observed no 

growth inhibition due to DNA damage in Escherichia 

coli treated with up to 4 mg/well scopolamine hydrobromide in the absence of S9 metabolic activation. 

In the second report, HeLa cells, treated for 5 hours 

with a 1 % solution of scopolamine HBr without S9, 

showed an increase in chromatid breaks 46 hours 

after the initiation of exposure (Vrba, 1967); HeLa 

cells analyzed 22 hours after the addition of scopolamine (1 %) to the culture medium showed no 

increase in the frequency ofchromatidbreaks. In 

addition, the author reported that no induction of 

chromatid breaks was noted in human leukocytes or 

BSC K cells treated in similar fashion, but no supporting data were included for this observation. 

The non-salt analogue, scopolamine, showedno 

evidence of mutagenic activity in Salmonella typhimurium strains TA98,TA100, or TA1537 with or 

withoutmetabolicactivation(Waskell, 1978; Glatt 

et al., 1983). A second structural analogue, scopolan, did not induce chromosome non-disjunctionor 

crossing-over in Aspergillus nidulans(Bignami et al., 

1974). 

STUDY RATIONALE 

Scopolamine hydrobromide trihydrate was nominated 

bytheNationalCancer Institute for toxicology and 

carcinogenicity testing because of considerable human 

exposure resulting from its use as a prescription or 

an over-the-counter drug and because it is a representativechemical from a class of alkaloids. Scopolamine is a suspect carcinogen because it contains an 

aliphatic epoxide moiety which may act as a biological alkylating agent. Its major use is in transdermal 

patches for the treatmentof motion sickness. Scopolamine hydrobromide trihydrate potentially could be 

used for the treatment of various ailments including 

acutemania, diarrhea, gastric and duodenal ulcers, 

gastrointestinal spasm, excessive salivation and 

sweating, and infantile cerebral palsy. It is also used 

as a mydriatic andcycloplegicagentandhasbeen 

used as an over-the-countersleeping aid. Thus 

human exposure occurs by several routes including 

oral, dermal, subcutaneous, andocular exposure. 

The route ofadministrationselected for the NTP 

studies was gavage because it mimics the oral exposure route in humans and higher doses can be administered to the animalthan can be achieved by the 

dosed feed route of administration. 
20 Scopolamine Hydrobromide Trihydrate,NTP TR 445 
21 

MATERIALS AND METHODS 

PROCUREMENT 

ANDCHARACTERIZATION 

OF SCOPOLAMINE HYDROBROMIDE 

TRIHYDRATE 

Scopolamine hydrobromide trihydrate was obtained 

in two lots, one from Rebeco Chemicals, Inc. 

(New York, NY; lot 14188), and one from Henley 

and Company, Inc. (New York, NY; lot 283). 

Lot 14188 was used during the 16-daystudies,and 

lot 283 was used during the 14-weekand2-year 

studies. Identity, purity, and stabilityanalyseswere 

conductedbytheanalyticalchemistry laboratory, 

MidwestResearch Institute (Kansas City, MO), 

(Appendix J). Reports on analyses performedin 

support of the scopolamine hydrobromide trihydrate 

studies are on file at the National Institute of Environmental Health Sciences (NIEHS). 

The chemical, a white powder,wasidentified as 

scopolamine hydrobromide trihydrate by infrared, 

ultraviolet/visible, and nuclearmagneticresonance 

spectroscopy. The purity ofboth lots was determinedbyelementalanalyses,Karl Fischer water 

analysis, functional group titration, thin-layer 

chromatography, high-performance and liquid

chromatography. Weight loss on drying was determined for lot 14188. 

For lot 14188, elemental analyses for carbon, hydrogen, nitrogen, and bromine were in agreement with 

the theoretical values for scopolamine hydrobromide 

trihydrate. Karl Fischer water analysisindicated 

10.8% f 0.3% water. Weightloss on drying 

indicated 10.8 % f 0.02 % water. Functional group 

titration indicated a purityof101 -9%f 0.1 % . Thin-layer chromatography by one system indicated 

a major spot, two traceimpurities,andoneslight 

trace impurity. Thin-layer by chromatography

another system indicated a major spot and one trace 

impurity. High-performance liquidchromatography 

revealed a major peak and one impurity with an area 

of 0.1% relative to the major peak. Major peak 

comparisons of lot 141 88 with a solution of dried 

UnitedStatesPharmacopeia XX (USP) reference 

standardscopolamine hydrobromide indicated that 

lot 14188 contained 88.8% f 0.2% scopolamine 

hydrobromide relative to the USP reference.

Lot 141 88 was determined to contain 89% scopolamine hydrobromide and 11 % water. The theoretical 

values for scopolamine hydrobromide trihydrate are 

87.7% scopolamine hydrobromide and 12.3 % water. 

For lot 283, elemental analyses for carbon, hydrogen, nitrogen, and bromine were in agreement with 

the theoretical values for scopolamine hydrobromide 

trihydrate. Karl Fischer water analysis indicated 

1 1.2 % f 0.2 % water. Functional group titration 

indicated a purity of 10 1.7 % f 0.6% . Thin-layer 

chromatography indicated a major spot and one trace 

impurity. High-performance liquid chromatography 

revealed a major peak and one impurity with an area 

of 0.2% relative to the major peak. Major peak 

comparisons of lot 283 with a solution of dried USP 

referencestandardscopolamine hydrobromide indicated that lot 283 contained 89.2% f 0.3% scopolamine hydrobromide relative to the USP reference. 

Major peak comparisons of lot 283 with lot 14188 

indicated a purity of 100.0% f 0.4% relative to 

lot 14188. Lot 283 was also determined to contain 

89% scopolamine hydrobromide and 1 1 % water. 

Stability studies of the bulk chemical were performed 

the analyticalby hemistry laboratory using

high-performanceliquid chromatography. These 

studiesindicatedthatscopolamine hydrobromide 

trihydrate was stable as a bulk chemical for 2 weeks 

when stored protected from light at room temperature 

in sealed containers under a nitrogen headspace. To 

ensure stability, the bulkchemicalwas stored in 

amberglass jars at approximately 25°C under a 

nitrogen headspace. 

Stabilitywasmonitoredby the study laboratory 

during the 16-day, 14-week, and 2-year studies using 

high-performance liquid chromatography and potentiometric titration (2-year studies). No degradation of 

the bulk chemical was detected. 
22 

PREPARATIONANDANALYSIS 

OF DOSEFORMULATIONS 

Doseformulationsforthe16-day and14-week 

studieswerepreparedevery two weeks.Forthe 

2-yearratstudy,doseformulationswereprepared 

weekly for thefirstfourmonthsevery 2 weeks 

thereafter.Doseformulationswerepreparedevery 

2 weeks throughout the 2-year mouse study. Formulations were prepared by mixing scopolamine hydrobromidetrihydratewithwater(Table51).Stability 

studies of the 1 1 and 0.2 mg/mL dose formulations 

were performed by theanalyticalchemistrylaboratory using high-performance liquid chromatography. 

The dose formulations, when stored at room tempera-and thymus were weighed. Complete histopathologic ture and protected from light, were stable for at least examinations were performed on all control rats and 3 weeks. 

Periodicanalysesofthedoseformulationsof 

scopolamine hydrobromide trihydrate were 

atthestudylaboratoryandanalyticalchemistry 

laboratory ultraviolethisible using spectrometry 

(16-dayand14-weekstudies) or high-performance 

liquidchromatography(2-yearstudies).Duringthe 

16-day studies, the formulations were analyzed at the 

beginning of the studies (Table 52). For the 14-week 

and2-yearstudies,theformulationswereanalyzed 

every 6 to 8 weeks(Tables53and54).Duringthe 

16-day studies, 70% (7/10) of the dose formulations 

were within 10% of the target concentration with no 

value differing more than 16 % from the target concentration. All ofthedoseformulationsanalyzed 

during the 14-week studies were within 10% of the 

target concentration. During the 2-year studies, 92% 

(72178) of the dose formulations were .within 10% of 

the target concentration with no value differing more12(rats) or 16 (mice)daysandwere 6 or 7 weeks than 20% from thetargetconcentration. For the old on the first day of the studies. Before the studies 14-weekand2-yearstudies,resultsofperiodic began, five male and five female rats and mice were referee analyses performed by the analytical chemistry laboratoryagreedwiththeresults ofthestudy 

laboratory (Table 55). 

16-DAY STUDIES 

Male and female F344/N rats and B6C3F, mice were 

obtained from Frederick Cancer Facility (Frederick, 

MD). Onreceipttheratsandmicewere29days 

old. Animals were quarantined for 12 days and were 

approximately 6 weeksoldonthefirstdayofthe 

study. Groups of five male and five female rats and 

Scopolamine Hydrobromide Trihydrate, NTP TR 445 

mice received scopolamine hydrobromide trihydrate 

in distilled water by gavage at doses of 0, 75, 150, 

300, 600, or 1,200 mg/kg body weight (rats) or 0, 

150,250,450,900, or 1,800 mg/kg(mice).Feed 

andwaterwereavailable ad libitum. Ratswere 

housed five per cage and mice were housed individually. Clinical findings were recorded twice daily for 

ratsandmice.Theanimalswereweighedinitially, 

weekly, and at the end of the studies. Details of the 

study design and animal maintenance are summarized 

in Table 2. 

A necropsy wasperformedonallanimals.The 

brain,heart,rightkidney,liver,lungs,righttestis, 

mice,andonratsreceiving 1,200mg/kgandmice 

receiving1,800mg/kg.Inaddition,thelivers of 

femaleratsreceiving600mg/kg,theliver of one

conductedmale rat receiving 75 mg/kg, and the livers, spleens, 

andthymusesofmaleandfemalemicereceiving 

900 mg/kg were examined. 

14-WEEK STUDIES 

The 14-week studies were conducted to evaluate the 

cumulativetoxiceffectsofrepeatedexposureto 

scopolaminehydrobromidetrihydrateandtodetermine the appropriate doses to be used in the 2-year 

studies. 

Male and female F344/N rats and B6C3F, mice were 

obtained from Simonsen Laboratories, Inc. (Gilroy , 

CA). On receipt, the rats were 4 weeks old and mice 

were 5 weeksold.Animalswerequarantinedfor 

randomlyselected for parasite evaluation and gross 

observationforevidenceofdisease.Attheend of 

thestudies,serologicanalyseswereperformedon 

five male and five female sentinel rats and mice using 

theprotocolsoftheNTPSentinelAnimalProgram 

(Appendix L). 

Groupsof 10 maleand10femaleratsandmice 

receivedscopolaminehydrobromidetrihydratein 

distilled water by gavage at doses of 0, 15, 45, 135, 

400, or 1,200 mg/kgbodyweight.Feedandwater 

were availablead libitum. Rats were housed five per 
Materials and Methods 23 

cage and mice were housed individually.Clinical 

findings andconsumption recorded feed were 

weekly. The animals were weighed initially, weekly, 

and at the end of the studies.Details of the study 

design and animalmaintenance are summarized in 

Table 2. 

At the end of the 1Cweek studies, blood for hematology was collected via the retroorbital sinus from rats 

and mice under carbon dioxide anesthesia. Samples 

wereplacedin containers containing EDTA as an 

anticoagulant. parameters Hematologywere 

measured on an Ortho ELT-8 hematology analyzer 

(Ortho Instruments, Westwood, MA).Differential 

leukocyte counts, morphologic evaluations of blood 

cellsand platelets, and reticulocytecountswere 

determinedby light microscopy of bloodfilms 

stainedwith a combination ofmethyleneblueand 

buffered Wright-Giemsa tain. The hematology 

parameters measured are listed in Table 2. 

Atthe end of the studies, samples for sperm morphologyandvaginalcytologyevaluationswere 

collected from rats receiving 0, 45, 135, or 

400 mg/kg and micereceiving 0, 135, 400, or 

1,200 mg/kg. The parameters evaluated are listed in 

Table 2. Methods used were those described in the 

NTP’s sperm andvaginalcytologyevaluations 

protocol (NTP, 1983). For 7 consecutive days prior 

to the scheduled terminal sacrifice, the vaginal vaults 

of the females were moistened with saline, if necessary, and aspirated samples of vaginal fluid and cells 

were transferred to slidesandstained.Relative 

numbers of leukocytes, nucleated epithelial cells, and 

large squamous epithelialcellsweredetermined to 

ascertain estrous cycle stage (i.e., diestrus, proestrus, 

estrus, and metestrus). All males were evaluated for 

sperm morphology, count, and motility. The right 

testisand right epididymis isolated were and 

weighed. The tail of the epididymis (cauda epididymis)was then removed from the epididymalbody 

(corpus epididymis) and weighed. Test yolk (rats) or 

modified Tyrode’s buffer (mice) was applied to slides 

and a small incision was made at the distal border of 

the cauda epididymis. The sperm effluxing from the 

incision were dispersed in the buffer on the slides, 

and the numbers of motile and nonmotile spermatozoawerecounted for five fields per slide by two 

observers. Followingcompletionof sperm motility 

estimates, each right cauda epididymis was placed in 

bufferedsaline solution and finelyminced. The 

tissuewasincubatedinthe saline solution and then 

heat fixed at 65 O C. Sperm density was then determined microscopically with the aid of a hemacytometer. Four sperm morphology slides were prepared 

for each animalevaluated. An aliquot of killed 

sperm suspension was stained in a test tube, spread 

on a microscope slide with coverslip, and examined. 

A necropsy was performed on all animals. The 

brain, heart, right kidney, liver, lungs, right ovary, 

right testis, thymus, and uterus wereweighed. 

Tissues for microscopic examination were fixed and 

preservedin 10% neutral buffered formalin, 

processedand trimmed, embeddedin paraffin, 

sectioned to a thickness of 5 to 6 pm, and stained 

withhematoxylinand eosin. Completehistopathologic examinations were performed on all control rats 

and mice, male rats receiving 135 mg/kg or higher, 

female rats receiving 400 mg/kg or higher, male and 

femalemicereceiving 1,200 mg/kg and one male 

mouse receiving 135 mg/kg that died early. Table 2 

lists the tissues and organs routinely examined. 

%YEAR STUDIES 

Study Design 

Groups of 60 male and 60 female rats and 70 male 

and 70 femalemicereceived scopolamine hydrobromide trihydrate in distilledwaterbygavage at 

doses of 0, 1,5,or 25 mg/kg body weight. Ten 

male and ten female rats and mice from each group 

received ophthalmic examinations during quarantine 

andat15months.Thesemicewere removed from 

the study following the 15 month examination without necropsy. The 10 maleand 10 female rats used 

for ophthalmic examination andan additional 10male 

and 10 female mice from each group were evaluated 

at15months for alterations in hematology, histopathology,and organ weights. The rats bled for 

hematologyanalyseswerebled again 1 hour after 

dosing for determination of scopolamine levels in 

plasma. 

Source and Specification of Animals 

Male and female F344/N rats and B6C3F, mice were 

obtained from Simonsen Laboratories (Gilroy, CA) 

for use in the2-yearstudies.Ratsandmice were 

quarantined for 14 days before the beginning of the 

studies. Five male and five female rats and mice 
24 

were selected for parasite evaluation and gross 

observation of disease. Serologysampleswere 

collected for viral screening. Ratsandmicewere 

approximately 6 weeksoldatthebeginningofthe 

studies. The health oftheanimalswasmonitored 

during the studies according to the protocols of the 

NTP Sentinel Animal Program (Appendix L). 

Animal Maintenance 

Rats were housed five per cage andmicewere 

housed individually. Feed andwaterwereavailable 

ad libitum. Feed consumption was measured for 

7 consecutive days each month by cage. Cages and 

racks were rotated every 2 weeks. Further details of 

animalmaintenance are given in Table 2. Information on feed composition and contaminants is provided in Appendix K. 

Clinical Examinations and Pathology 

Allanimalswere observed twice daily. During the 

first 13 weeks, clinical findings were recorded every 

three to four weeks for rats and every three to six 

weeks for mice. Clinical findings were recorded 

monthly thereafter. Bodyweightswererecorded 

weekly for 13 weeks, monthly thereafter, and at the 

end of the studies. Before the studies began, 10 male 

and 10 female rats and mice per group were randomlyselected for ophthalmic examination.These 

animals were examined again at 15months. No 

mydriatic agent was used to facilitate theexaminations, Funduscopic examination was accomplished 

using an ophthalmoscope; examinations of the anterior chamber were performed using a slit-lamp biomicroscope. Mice that received ophthalmicexams 

were discarded without further evaluation. 

At the 15-month interim evaluation, within the hour 

after dosing, blood for hematology was collected via 

the retroorbital sinus of rats and miceanesthetized 

with a mixture of carbon dioxide and oxygen. 

Because of a flooding incident that killed 16 female 

rats receiving 1 mg/kg, no females in this dose group 

were bled or necropsied at 15 months. The hematology parameters measured are listed in Table 2; 

methods used were similar to those used inthe 

14-week studies. Additionally,the rats bled at 

15 months for hematology analyseswerebled an 

hour after dosing by cardiac puncture under CO,:O, 

anesthesia for plasma scopolaminedetermination 

Scopolamine Hydrobromide Trihydrate, NTP TR 445 

studies.Ratsandmicebled for hematology at the, 

15-month interim evaluation were necropsied. 

Bloodsamplescollected for plasma scopolamine 

determinationsweresent to Midwest Research 

Institute(Kansas City, MO). Analyseswas performed using gas chromatography and mass spectrosCOPY* 

A complete necropsyand microscopic examination 

were performed on all core study animals. At the 

15-monthinterim evaluations the right epididymis, 

right kidney, liver, and right testis of rats and mice 

wereweighed(excluding female rats receiving 

1 mg/kg). At necropsy, all organs and tissues were 

examined for grossly visible lesions. Major tissues 

werefixedand preserved in 10 % neutral buffered 

formalin, processed and trimmed, embeddedin 

paraffin, sectioned to a thickness of 5 to 6 pm, 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 2. 

Additional groups of 10 male and 10 female rats 

receivingscopolamine hydrobromide trihydrate in 

waterbygavage at doses of 0, 1, 5, or 25 mg/kg 

body weight were administered neurobehavioral tests 

prior to the study, on the first day of study, and after 

3, 6, 9, 12, and 24 months of dosing. The same 

animals were used at each time point. These neurobehavioraltestsincluded motor activity, grip 

strength, thermal sensitivity, startle responsiveness, 

andpassiveavoidance. Further details of these 

evaluations are outlined in Appendix I. 

Microscopic evaluations were completed by the study 

laboratory pathologist, and the pathology data were 

entered into the ToxicologyData Management 

System. The microscopic 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 liver, 
Materials and Methods 25 

pituitary gland, and spleen of male and female rats, 

the parathyroid gland, thyroid gland, and stomach of 

male rats, and the ovaries of female rats. In mice, 

the liver, forestomach, thyroid gland, lung, and 

kidney of males and females,'teeth, pancreatic islets, 

and preputial gland of males, and the pituitary gland, 

uterus, andbonemarrowoffemalemicewere 

examined. 

The quality assessment report and the reviewed slides 

weresubmitted to the NTP PathologyWorking 

Group (PWG) chairperson, who reviewed the selectedtissuesandaddressedanyinconsistenciesinthe 

diagnoses made by the laboratory and quality assessment Representative pathologists. histopathology 

slides containing examples of lesionsrelated to 

chemical administration, examples of disagreements 

indiagnosesbetweenthe laboratory and quality 

assessment pathologists,or lesions of general interest 

were presented by the chairperson to the PWG for 

review. The PWG consistedofthequalityassessment pathologist and other pathologists experienced 

in rodent toxicologic pathology. This group examinedthetissueswithoutanyknowledgeof dose 

groups or previously rendered diagnoses. When the 

PWGconsensus differed from the opinion of the 

laboratory pathologist, the diagnosiswaschanged. 

Thus, the final diagnosesrepresent a consensus of 

qualityassessmentpathologists,thePWGchairperson, and the PWG. Detailsofthesereviewprocedures have been described, in part, by Maronpot and 

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

subsequentanalysesofthepathologydata,the 

diagnosed lesions for each tissue type were evaluated 

separately or combined according to the guidelines of 

McConnell et aZ. (1986). 

STATISTICALMETHODS 

Survival Analyses 

The probability of survival was estimatedbythe 

product-limit procedure of Kaplan and Meier (1958) 

and is presented in the form of graphs. Animals 

found dead of other than natural causeswerecensored from the survival analyses; animals dying from 

natural causes were not censored. Statistical analyses 

for possible dose-related effects on survivalused 

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 Al, A5, B1, B5, C1, C5, Dl, 

and D5 are given as the number of animals bearing 

suchlesionsat a specific anatomic' site and the 

numberofanimalswith that site examined microscopically. For calculation of statistical significance, 

theincidencesofmostneoplasms (Tables A3, B3, 

C3, and D3) and all nonneoplastic lesions are given 

as the ratio of the number of affected animals to the 

number of animals withthe site examined microscopically. However, when macroscopic examination 

wasrequired to detect neoplasmsin certain tissues 

(e.g., harderian gland, intestine, mammary glands 

andskin) before microscopic evaluation, or when 

neoplasms had multiple potential sites of occurrence 

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

consist of the numberofanimals on which a necropsywas performed. Tables A3, B3, C3, and D3 

also give the survival-adjusted neoplasmrate for each 

group and eachsite-specific neoplasm, i.e., the 

Kaplan-Meierestimateofneoplasmincidence that 

would have been observed at the end of the study in 

theabsenceofmortality from all other competing 

risks (Kaplan and Meier, 1958). 

Analysis of Neoplasm Incidences 

Themajorityofneoplasmsinthese studies were 

considered to be incidental to the cause of death or 

notrapidlylethal. 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 

andthus did not affectthe risk of death. In this 

approach,neoplasmprevalencewasmodeled as a 

logisticfunction of chemical exposure and time. 

Both linear and quadratic terms in time were incorporated initially, and the quadratic term was eliminatedif the fit ofthemodelwas not significantly 

enhanced. The neoplasm incidencesofdosedand 

control groups were compared on the basis of the 

likelihood score test for the regression coefficient of 

dose. This method of 'adjusting for intercurrent 

mortalityistheprevalenceanalysisofDinseand 

Lagakos (1983), further described and illustrated by 

DinseandHaseman (1986). Whenneoplasms are 

incidental, this comparison of thetime-specific 
26 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

neoplasm prevalences also provides a comparison of to determine whether a trend-sensitive test (Williams’ 

the time-specific neoplasm incidences (McKnight andor Shirley’s test) was more appropriate for pairwise 

Crowley,1984). 

Inaddition to logistic regression, other methods of 

statistical analysis were used, and the resultsof these 

testsaresummarizedintheappendixes.These 

methodsincludethelifetabletest(Cox,1972; 

Tarone,1975),appropriate for rapidly lethalneoplasms,and theFisherexacttestand theCochranArmitagetrendtest(Armitage,1971;Gart et al., 

1979), procedures based on the overall proportion of 

neoplasm-bearing animals. 

Tests of significance included pairwise comparisons 

of each dosed group with controls and a test for an 

overall dose-related trend. Continuity-corrected tests 

were used in the analysisof neoplasm incidence, and 

reportedPvaluesareonesided.Theprocedures 

comparisonsthanatesthatdoesnotassumea 

monotonicdose-relatedtrend(Dunnett’s or Dum’s 

test). Prior to analysis, extreme values identified by 

theoutliertest of Dixon andMassey(1951)were 

examined by NTP personnel, and implausible values 

were eliminated from the analysis. Average severity 

valueswereanalyzed for significance usingthe 

Mann-Whitney U test (Hollander and Wolfe, 1973). 

Becausevaginalcytologydataareproportions(the 

proportions of the observation period that an animal 

was in a given estrous state), an arcsine transformation was usedtobringthedataintocloserconformancenormality Treatment with assumptions. 

effects were investigated by applying a multivariate 

analysis of variance (Morrison, 1976) to the transformeddatatotest for simultaneous equality of 

measurements across dose levels. 

described in the preceding paragraphs were also used 

toevaluate nonneoplastic For selected lesions. Historical Control Data

further discussion of these statistical methods, refer Although the concurrent control group is always the 

to Haseman (1984). firstandmostappropriatecontrolgroupused for 

evaluation, historical controldatacanbehelpful in 

Analysis of Nonneoplastic Lesion Incidences theoverallassessmentofneoplasmincidencein 

Because all nonneoplastic lesions in this study were certain instances. Consequently, neoplasm incidences 

considered to be incidental to the cause of death or fromtheNTPhistoricalcontroldatabase,whichis 

not rapidly lethal, the primary statistical analysis usedupdated yearly, are included in the NTP reports for 

was alogisticregressionanalysisin whichnon- neoplasmsappearingtoshowcompound-related 

neoplastic lesion prevalence was modeled as a logisticeffects. 

function of chemical exposure and time. For lesions 

detectedattheinterimevaluation,theFisherexact 

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 comparisonsbetweendosedand 

controlgroupsintheanalysis of continuous variables.Organandbodyweightdata, whichhave 

approximatelynormaldistributions,wereanalyzed 

using the parametric multiple comparison procedures 

of Dunnett(1955)andWilliams(197 1, 1972). 

Hematologyepididymal data, and spermatozoal 

whichavetypicallyskewedistributions,were 

analyzed using the nonparametric multiple comparisonmethods of Shirley (1977)andDunn(1964). 

Jonckheere’stest(Jonckheere,1954) wasusedto 

assess the significance of the dose-related trends and 

QUALITYASSURANCEMETHODS 

The14-weekand2-yearstudieswereconducted 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 

submittedtotheNTPArchives,thesestudieswere 

auditedretrospectively by anindependentquality 

assurance Separate covering contractor. audits 

completenessandaccuracy of the pathologydata, 

pathologyspecimens,finalpathologytables,anda 

draft of this NTP Technical Report were conducted. 

Auditproceduresandfindings are presented in the 

reports and are on file at NIEHS. The audit findings 

werereviewedandassessedbyNTP 

commentshadbeenresolved 

addressedduringthepreparation 

Report. 

staff, so all 

or wereotherwise 

of this Technical 
Materials and Methods 27 

GENETICTOXICOLOGY 

Thegenetictoxicity of scopolamine hydrobromide 

trihydrate was assessed by testing the ability of the 

chemical to induce mutationsinvariousstrains of 

Salmonella typhimurium, sister chromatid exchanges 

andchromosomalaberrationsinculturedChinese 

hamster ovary cells, and increases in the frequency of 

micronucleatederythrocytesinmouseperipheral 

blood. The protocols for these studies and the results 

are given in Appendix E. 

Thegenetictoxicitystudiesofscopolaminehydrobromide trihydrate are part of a larger effort by the 

NTP to develop adatabasethatwouldpermitthe 

Thereisastrongcorrelationbetweenachemical’s 

potentialelectrophilicity(structuralalert to DNA 

reactivity), mutagenicity in Salmonella, and carcinogenicityinrodents.Thecombinationofelectrophilicity.and Salmonella mutagenicityishighly 

correlatedwiththeinductionofcarcinogenicityin 

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

andTennant, 1991). Other in vitro genetictoxicity 

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 

evaluation of carcinogenicity in experimental animalscarcinogenicity (89% ofthe Salmonella mutagens 

from the structure and responses of the chemical in were rodent carcinogens), and that there is no comshort-term in vitro and in vivo genetic toxicity tests. plementarity among thein vitro genetic toxicity tests. 

These genetic toxicity tests were originally developedThat is, nobattery of teststhatincludedthe 

to study mechanismsofchemicallyinducedDNA 

damageand to predict carcinogenicityinanimals, 

based on the electrophilic theoryof chemical carcinogenesis and the somatic mutation theory (Miller and 

Miller, 1977; Straw, 1981; Crawford, 1985). 

Salmonella testimprovedthepredictivityofthe 

Salmonella test alone.Thepredictivity for carcinogenicityof apositiveresponseinbonemarrow 

chromosome aberration or micronucleus tests is not 

yet defined. 
28 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLE2 

Experimental Design and Materials and Methods in the GavageStudia. 

of Scopolamine Hydrobromide Trihydrate 

16-Day Studies 

Study Laboratory 

Southern Research Institute 

(Birmingham, AL) 

Strain and Species 

Rats: F344lN 

Mice: B6C3F, 

Animal Source 

Frederick Cancer Facility 

(Frederick, MD) 

Time Held Before Studies 

12 days 

Average Age When Studies Began 

6 weeks 

Date of First Dose 

Rats: 1-2 July 1985 

Mice: 8-9 July 1985 

Duration of Dosing 

16 days 

Date of Last Dose 

Rats: 17-18July1985 

Mice: 23-24 July 1985 

Necropsy Dates 

Rats: 18-19 July 1985 

Mice:24-25July1985 

Average Age at Necropsy 

8 weeks 

14-Week Studies 

Southern Research Institute 

(Birmingham, AL) 

Rats:F344lN 

Mice: B6C3F, 

Simonsen Laboratories, Inc. 

(Gilroy, CA) 

Rats: 12 days 

Mice:16days 

Rats: 6 weeks 

Mice: 7 weeks 

Rats: 17March1986 

Mice: 21 March 1986 

14 weeks 

Rats: 17-19 June 1986 

Mice: 24-26 June 1986 

Rats: 18-20 June1986 

Mice: 25-27 June 1986 

Rats: 19-20weeks 

Mice:20-21weeks 

2-Year Studies 

Battelle Columbus Laboratories 

(Columbus. OH) 

Rats: F344lN 

Mice: B6C3F, 

Simonsen Laboratories, Inc. 

(Gilroy. CA) 

14 days 

6 weeks 

Rats: 20 October 1988 

Mice: 22 September 1988 

Rats: 104weeks 

Mice:104-105weeks 

Rats: 17 October 1990 

Mice: 20 September 1990 

Rats:15-monthinterimevaluation - 18 January (males)-19 January 

(females)1990 

terminal sacrifice -15-17 October 

1990 

Mice:15-monthinterimevaluation -

21-22 December 1990 

terminal sacrifice -17-21 September 

1990 

Rats: 110 weeks 

Mice:111weeks 
Materials and Methods 29 

TABLE2 

Experimental Design and Materials and Methods in the Gavage Studies 

of Scopolamine Hydrobromide Trihydrate (continued) 

16-Day Studies 

Size of Study Groups 

5 males and 5 females 

Method of Distribution 

Animals were distributed randomly into 

groups of approximately equal initial 

mean body weight. 

Aniials per Cage 

Rats: 5 

Mice: 1 

Method of Animal Identification 

Toe clip 

Diet 

NIH-07 open formula pelleted diet 

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

available ad libitum 

WaterDistribution 

Tap water (City of Birmingham 

Municipal supply) via automatic 

watering system (Edstrom Industries, 

Waterford, WI), available ad libirum 

Cages 

Polycarbonate (Lab Products Inc., 

Maywood. NJ), changed twice weekly 

Bedding 

BetaChips" (Northeastern Products 

Co., Warrensburg, NY) 

14-WeekStudies 

10 males and 10 females 

Same as 16day studies 

Rats: 5 

Mice: 1 

Toe clip 

Same as 16day studies 

Same as 16day studies 

Same as 16day studies 

Same as 16day studies 

2-Year Studies 

15-Month interimRats: 10 males and 10 females 

received ophthalmic examinations, 

were bled for hematology and plasma 

scopolamine levels, and necropsied 

for histological examination and 

organ weights 

Mice: 10 males and 10 females were 

bled for hematology and necropsied 

for histological examination and 

organ weights 

10 males and 10 females received 

ophthalmic examinations and were 

discarded without necropsy 

Terminal -50 male and 50 female 

rats and mice 

Sameas 16day studies 

Rats: 5 

Mice: 1 

Rats: Tail tattoo 

Mice:Toe clip 

Same as 16day studies 

Tap water (Columbus Municipal 

Supply) via automatic watering system 

(Edstrom Industries, Waterford, WI), 

available ad libitum 

Same as 16day studies 

Sani-Chip" hardwood chips (P.J. 

Murphy Forest Products Corp., 

Montville. NJ) 
30 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLE2 

Experimental Design and Materials and Methods in the GavageStudies 

of Scopolamine Hydrobromide Trihydrate(continued) 

16-Day Studies l4Week Studies 2-Year Studies 

Cage Filters 

Fiber fiters Reemay' spun-bonded polyester DuPont 2024 spun-bonded polyester 

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

Racks 

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

Maywood, NJ),rotated once every Maywood, NJ)rotated once every 

2 weeks 2 weeks 

Animal Room Environment 

Temperature: 20.0' to 24.0" C (rats); Temperature: 20.0" to 25.0" C (rats); Temperature: 20.7" to 23.3" C (rats); 

21.0" to 24.0" C (mice) 20.0" to 24.90 C (mice) 20.0" to 24.4" C (mice) 

Relative humidity: 43 96 to 5746 (rats); Relativehumidity: 45% to 66% (rats); Relativehumidity: 24% to 72% (rats); 

45% to 55% (mice) 45% to 71% (mice) 305% to 73% (mice) 

Fluorescentlight: 12 hourslday Fluorescentlight: 12 hourslday Fluorescentlight: 12 hourslday 

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

10 changesmour 10 changeslhour 10 changesmour 

Doses 

Rats: 0.75, 150, 300. 600, or 0, 15. 45, 135, 400, or 1,200 mg/kg 0. 1, 5. or 25 mglkg 

1,200 mglkg 

Mice: 0. 150, 250, 450, 900, or 

1,800 mglkg 

Type and Frequency of Observation 

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

weighed initially, weekly, and at the weighed initially, weekly, and at the observations were recorded every 

end of the studies. Clinical findings endofthestudies.Clinical 3 4 weeks (rats) or 3-6 weeks (mice) 

were recorded twice daily. observations were recorded weekly. during the first 13 weeks and monthly 

Feed consumption was measured thereafter; body weights were recorded 

weekly by cage. weekly through week 13, monthly 

thereafter, and at the end of the 

studies.Feedconsumptionwas 

measured by cage for 7 consecutive 

days each month. 

Method of Sacrifice 

CO, asphyxiation CO, asphyxiation COz asphyxiation 

Necropsy 

All animalswerenecropsied.Organs All animalswerenecropsied.Organs All core-studyanimalswere 

kidney,liver,lungs,righttestis,andkidney,liver,lungs,rightovary,right15­

weighedincludedthe brain,heart,right 

monthinterimevaluationswerethe 

weighedincludedthe brain,heart, rightnecropsied.Organsweighed at the 

and thymus, testis, thymus. 

and right testis (excluding female rats 

receiving 1 mglkg). 
Materials and Methods 31 

TABLE2 

Experimental Design and Materials and Methodsin the Gavage Studies 

of Scopolamine Hydrobromide Trihydrate(continued) 

16-Day Studies 

Cliiical Pathology 

None 

Histopathology 

Complete histopathologic examinations 

were performed on all control rats and 

mice, as well as rats receiving 

1,200 mglkg and mice receiving 

1,800 mglkg. In addition to gross 

lesions, tissue masses and associated 

lymph nodes, the tissues examined 

included: adrenal gland, bone and 

marrow, brain, clitoral gland (rats) 

esophagus, gallbladder (mice), heart, 

kidney, large intestine (cecum, colon, 

and rectum), small intestine 

(duodenum, jejunum, and ileum), liver, 

lung, lymph nodes (mandibular and 

mesenteric), mammary gland, nose, 

ovary, pancreas, parathyroid gland, 

pituitary gland, preputial gland (rats), 

prostate gland, salivary gland, skin, 

spleen, stomach (forestomach and 

glandular stomach), testis with 

epididymis and seminal vesicle, 

thymus, thyroid gland, trachea, urinary 

bladder, and uterus. The liver of 

female rats receiving 600 mglkg, the 

liver of one male receiving 75 mglkg, 

and the liver, spleen, and thymus of 

male and female mice receiving 

900 mglkg were also examined. 

14-Week Studies 

At the end of the 14-week studies, 

blood was collected from the retroorbital sinus of rats and mice. 

Hematology: hematocrit; hemoglobin; 

erythrocyte, reticulocyte, and nucleated 

erythrocyte counts (rats); mean cell 

volume; mean cell hemoglobin; mean 

cell hemoglobin concentration; platelet 

count; and leukocyte count and 

differentials. 

Complete histopathologic examinations 

were performed on all control rats and 

mice, male rats receiving 135 mglkg or 

higher, female rats receiving 

400mglkg or higher, male and female 

mice receiving 1,200 mglkg and one 

male mouse receiving 135 mglkg that 

died early. In addition to gross 

lesions, tissue masses and associated 

lymph nodes, the tissues examined 

included: adrenal gland, bone and 

marrow, brain, clitoral gland (rats), 

esophagus, gallbladder (mice), heart, 

kidney, large intestine (cecum, colon, 

and rectum), small intestine 

(duodenum, jejunum, and ileum), liver, 

lung, lymph nodes (mandibular and 

mesenteric), mammary gland, nose, 

ovary, pancreas, parathyroid gland, 

pituitary gland, preputial gland (rats), 

prostate gland, salivary gland, skin, 

spleen, stomach (forestomach and 

glandular stomach), testis with 

epididymis and seminal vesicle, 

thymus, thyroid gland, trachea, urinary 

bladder. and uterus. 

2-Year Studies 

At the 15-month interim evaluation, 

blood for hematology was collected 

from the retroorbital sinus of rats and 

mice under CO,:O, anesthesia 

(excluding female rats that received 

1 mglkg). The hematology parameters 

measured were the same as those in the 

14-week studies. 

Complete histopathologic examinations 

were performed on all core-study 

animals. In addition to gross lesions, 

tissue masses, and associated lymph 

nodes, the tissues examined included: 

adrenal gland, bone and marrow, 

brain, clitoral gland, esophagus, 

gallbladder (mice), heart, kidney, large 

intestine (cecum, colon, and rectum), 

small intestine (duodenum, jejunum, 

and ileum), liver, lung, lymph nodes 

(mandibular or mesenteric), mammary 

gland, nose, ovary, pancreas, 

parathyroid gland, pituitary gland, 

preputial gland, prostate gland, salivary 

gland, spleen, stomach (forestomach 

and glandular stomach), testis with 

epididymis and seminal vesicle, 

thymus, thyroid gland, trachea, urinary 

bladder. and uterus. 
32 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE2 

Experimental Design and Materials and Methods in the Gavage Studies 

of Scopolamine Hydrobromide Trihydrate (continued) 

16-Day Studies 14-WeekStudies 

Sperm Morphology and Vaginal Cytology Evaluations 

None SpermvaginalsamplesNone and fluid were 

evaluated in 0, 45, 135, and 400 mglkg 

rats and 0, 135. 400 and 1,200 mglkg 

mice at the endof the studies. The 

parameters evaluated in males were 

sperm count, morphology, and 

motility.Theright cauda, right 

epididymis, and right testis were 

weighed.Vaginal fluid sampleswere 

collected for up to 7 consecutive days 

prior to the end of the studies for 

vaginalcytologyevaluations.The 

parameters evaluated in females were 

relative frequency of estrous stages and 

estrous cycle length. 

Ophthalmic Examination 

None None 

Plasma Scopolamine Determination 

None None 

Neurobehavioral Studies 

None None 

2-Year Studies 

10 male and 10 female rats and mice 

per dose group received ophthalmic 

examinations at the beginning of the 

study and again at the 15-month 

interim evaluation. 

At 15 months, 10 male and 10 female 

rats were bled for plasma scopolamine 

determinations. (Rats had been bled 

one hour earlier for hematology 

evaluations.) 

Ten male and ten female rats per dose 

group were administered 

neurobehavioral tests prior to the 

study, on the first day of study, and 

after 3, 6, 9, 12, and 24 months of 

exposure.Thesetestsincluded 

evaluations of motor activity, grip 

strength, thermal sensitivity, startle 

responsiveness, and passive avoidance. 
33 

RESULTS 

RATS Clinical findings included bilateral pupillary dilation 

16-DAY STUDY in all dosedanimalsandred eyelids in males and 

All rats survived to the end of the study (Table 3). femalesreceiving 1,200 mg/kg. No biologically 

The final mean body weights and body weight gains significant changes in organ weights were observed 

ofmalesreceiving 600 and 1,200 mg/kgandthe (Table Fl). There were no biologically significant 

mean body weight gain of males receiving300 mg/kg treatment-related gross or microscopic lesions, and 

weresignificantly lower than those of the control thehigh dose selected for the 14-weekstudywas 

group. 1,200 mg/kg, the same as that in the 16-day study. 

TABLE3 

Survival and Body Weights of Rats in the 16-Day Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Mean Body Weightb (g) Final Weight 

Dose Initial Survivala Final Change Controls Relative to

(mgW (W 

Male 

600 

300 

150 

75 

0 

1,200 515 

515 

515 

515 

515 

515 

125 f 7 

116 f 3 

118 f 4 

116 f 2 

119 f 4 

116 f 3 

166 f 5** 

166 f 6** 

173 f 4 

174 f 6 

175 f 5 

186 f 2 

41 f 8** 

50 f 5** 

56 f 2* 

58 f 4 

55 f 2 

69 f 1 

* 

90 

89 

93 

94 

94 

Female 

0 515 103 f 4 140 f 5 37 f 2 

75 515 103 f 4 133 f 5 30 f 6 95 

150 515 105 f 4 134 f 4 29 f 1 96 

300 515 102 f 3 134 f 4 31 f 1 96 

600 515 105 f 2 138 f 3 32 f 1 98 

1,200 515 105 f 2 135 f 3 31 f4 97 

* Significantly different (P 50.05)from the control group by Williams’ or Dunnett’s test 

** P50.01 

a Number of animals survivinglnumber initially in group 

Weights and weight changes are given as mean f standard error. 
34 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

14-WEEK STUDY Hyperactivity was visually observed in a few dosed 

Onefemalereceiving45mg/kg,onemaleandone males and females (males:0 mg/kg, 0/10;15 mg/kg, 

femalereceiving 135 mg/kg,sixmalesandone 0/10;45 mg/kg, 1/10;135 mg/kg, 1/10;400 mg/kg, 

femalereceiving400mg/kg,andeightmalesand 0/10;1,2OO-mg/kg,l/iO;females: 0/10,2/10,5/10, 

Hypoactivitywasalsovisually seven females receiving1,200mg/kg died during the 3/10, 1/10, 1/10). 

study(Table4).Thefinal meanbodyweightsand observedinsomeotherdosedmalesandfemales 

meanbodyweightgainsofalldosedmalesand (males: 0/10, 0/10, 0/10, 0/10, 3/10, 3/10; females:

andincreased femalesweresignificantlylowerthanthoseofthe 0/10, 0/10, 0/10, 1/10, 1/10, 5/10) 

controlgroups.Clinicalfindingsincludedbilateral with increasing dose. 

pupillary dilation in all dosed males and females and 

reddeningoftheeyesin 15 mg/kgmalesand 135, Thehematologydata for rats in the14-week study 

400, and 1,200mg/kg males and females. arelistedinTable G1. Hematocrit,hemoglobin 

TABLE 4 

Survival and Body Weights of Rats in the 14-Week Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Mean Body Weightb (g) Final Weight

Dose Initial Survivala Final Change Relative to Controls

Male 

0 10110 116 f 3 341 f 6 225 f 5 

15 10110 116 f 3 312 f 7** 197 f 6** 92 

45 10110 115 f 3 302 f 4** 188 f 5** 89 

135 8/10’ 114 f 3 300 f 7** 186 f 5** 88 

400 4/1Od 115 f 2 304 f 3** 188 f 7** 89 

1,200 2/10e 116 f 2 277 f 37** 163 f 30** 81 

Female 

0 10110 96 f 1 203 f 2 107 f 2 

15 10110 97 f 1 193 f 2** 96 f 2** 95 

45 9/1Of 98 f 2 196 f 2* 97 f 1** 97 

135 9ll@ 95 f 1 190 f 2** 96 f 3** 94 

400 9/10f 96 f 1 189 f 2** 93 f 3** 93 

1,200 311$ 95 f 1 192 f 6** 95 f 5** 94 

* Significantly different(PsO.05) from the control group by Williams’ or Dunnett’s test 

** PSO.01 

Number of animals survivinglnumber initially in group. Subsequent calculations are based on animals surviving to the end of the study 

Weights and weight changes are given as mean f standard error. 

Week of death: 7, 14 

Week of death: 1, 2,7, 7,13, 13 

Week of death: 1, 1, 1, 3, 4, 5, 9, 14 

Week of death: 4 

Week of death: 8 

Week of death: 1, 2,5, 6, 7, 7, 10 
Results 35 

concentration, and/or erythrocyte count in male and thymus weights of 15, 45, 135, and 400 mg/kg males 

female rats receiving 45 mg/kg or greater wereandfemalesand 1,200 mg/kg females were signifislightly higher than those of the control groups. In cantlylowerthanthoseof the control groups 

general, these differences were most prominent in the (Table F2). 

400 and 1,200 mg/kg groups. Higherhematocrit, 

hemoglobin concentration, and erythrocyte count 

would be consistent with thehemoconcentration 

associatedwith dehydration (relative erythrocytosis). 

The mortality and lower body weights of the dosed 

animals suggest that thedosed rats did noteat or 

drink as much as the controls and some dehydration 

wouldhave occurred. A minimal to mild mature 

neutrophilia, evidenced by higher segmented neutrophil numbers than those in the control group, occurred in all dosed male rats. Neutrophiliaisoften 

a result of an increased tissue demand for granulocytes due to inflammation. There was, however, no 

microscopicevidenceofinflammationthatcould 

account for the neutrophilia. Thus, other mechanisms that alter granulopoiesis and/or rate of release 

from the bone marrow, redistribution of neutrophils 

betweenthemarginalandthecirculating pools, or 

increase the intravascular neutrophil life span could 

be considered. Other hematology differences were 

Sperm morphology andvaginal cytology parameters 

in dosed rats were similar to those in the control 

groups (Table HI). 

Some males (0/10, 0/10, 0/10, 2/10, 3/10, 4/10)and 

females (0/10, 0/10, 1/10, 1/10, 0/10, 3/10) died 

from esophageal obstructions consisting of feed and 

bedding material in the posterior pharynx. Tracheal 

obstruction occurred concurrently with esophageal 

obstruction as a result of food build-up in the oropharyngealregion. This condition is considered to be 

secondary to the inhibitory effects of scopolamine 

hydrobromide trihydrate on salivary gland secretions 

and on esophageal smooth muscle involved in swallowing. There were no other significant treatmentrelated gross or microscopic findings. 

Dose SelectionRationale: Based on lower mean 

sporadic and werenottreatmentrelated.bodyweightsinalldosedanimalsand lower survival 

in 400 mg/kg males and 1,200 mg/kg males and 

The absolute andrelative liver weightsofemalescompared to body weightsand survival in the 

1,200 mg/kg females weresignificantly greater than control groups, doses selected for the 2-year studies 

thoseof the control group. The absoluteandrelativewere 1, 5, and 25 mg/kg. 
36 

%YEAR STUDY 

Survival 

Estimates of 2-year survivalprobabilities for male 

andfemalerats 'are shown inTable 5 andinthe 

Kaplan-Meier survival curves (Figure1). Survival of 

femaleratsreceiving 1 and 25mg/kgwas significantlylowerthanthatofthecontrolgroup.The 

lower survival in 1 mg/kg females was due primarily 

to a cage flooding accident resulting froma malfunctioning automatic watering system valve. 

Body Weights, Clinical Findings, 

and Ophthalmic Examination Findings 

Meanbodyweightsof 1 and 5 mg/kgmalesand 

females were similarto those of the controls throughout the study (Figure 2; Tables 6 and 7). The mean 

bodyweightof25mg/kgmaleswasslightlylower 

than that of the control group from about week 25 

through week 97. Themeanbodyweightof 

25 mg/kg females was lower than that of the control 

Scopolamine Hydrobromide Trihydrate, NTP TR 445 

Compared to controls, hematocrit was slightly higher 

in the 25 mg/kg male rats, similar to effects observed 

in the 14-week study; this would be consistent with 

dehydrationresultinginhemoconcentration. The 

reticulocytecountin25mg/kgfemaleratswas 

slightlylowerthanthatinthecontrol group. This 

result is consistent with the lower body weights, and 

thus a decreased nutritional status, exhibited by these 

animals. 

Plasma Scopolamine Determinations 

Serumsamplescollectedfromrats 1 hour after 

dosingwereanalyzed to determine serumscopolamineconcentrations.Serumscopolamineconcentrationanalysesindicated 6 ng scopolamine/mL 

serum for the 5 mg/kgfemalesampleand12and 

28ng/mL for the25mg/kgmaleandfemale 

samples,respectively.Theamounts of scopolamine 

in the other serum samples were below the minimum 

detection limit (4 ng/mL) of the analysis method. 

group from week 25, and the final mean body weight 

was 81% thatofthecontrols.Clinicalfindings Neurobehavioral Findings 

included bilateral pupillary dilation in all dosed males Horizontal motor activity of 25 mg/kg females was 

andfemales.Ophthalmicexaminationrevealedno 

significant findings. 

Hematology 

The hematology data for rats at the 15-month interim 

evaluation in the 2-year study are listed in Table G2. 

significantly greater than that of the control group on 

days 90, 180, and 360. The startle response of 5 and 

25 mg/kg females was significantly lower than thatof 

thecontrolgrouponday90.Onday180,passive 

avoidance by 25 mg/kg males was significantly lower 

than that by the control group (Appendix I). 
Results 37 

TABLE5 

Survival of Rats in the 2-YearGavage Study of Scopolamine Hydrobromide Trihydrate 

Vehicle Control 1 mgkg 5 mgkg 25 mgkg 

Male 

in initially Animals 60 60 60 60 

15-Month interim evaluationa 10 10 10 10 

Accidental deathsa 2 2 1 1 

Moribund 23 30 20 17 

Natural deaths 5 4 7 4 

Animals surviving to study termination 20 14 22 28 

Percent probability of survival at the end of studyb 42 30 45 58 

Mean survival (days)' 625 610 632 620 

lysisd Survival P=O.219 P=0.995N P=0.274N 

Female 

in initially Animals 60 60 60 60 

15-Month interim evaluationa 10 0 10 10 

Accidental deathsa 0 18e 2 5 

Moribund 13 10 6 7 

Natural deaths 3 15 16 16 

Animals surviving to study termination 34 17 26 22 

Percent probability of survival at the end of study 69 41 56 53 

Mean survival (days) 643 476 622 539 

Survival analysis P=O.311 P=O.o02 P=O.286 P=O.o41 

a Censoredfromsurvivalanalyses 

Kaplan-Meier determinations ' Mean of all deaths (uncensored, censored, and terminal sacrifice). 

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

1972) with the controls are in the dosed columns. A negative trend or lower mortality in a dose group is indicated by N. 

e Sixteen deaths were due to a cage floodingaccident 
38 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

1.0 

0.9 

2 

2

m 

0.5 

0 

a 

cc 

0.4 

0.3 

0.2 -~I ~~~ I 

15 ;0 45 60 

WEEKS ON STUDY 

1.0 

0.9 

0

L 

d 

Oe6 

m

2 0.5 

8 

a 0.4 

0.5 

0.2 

;5 90 

15 30 45 60 75 90 1C 

WEEKS ON STUDY 

FIGURE 1 

Kaplan-Meier Survival Curves for Male and Female Rats Administered 

Scopolamine Hydrobromide Trihydrate in Water by Gavage for 2 Years 
Results 39 

15 SO 45 60 75 90 109 

WEEKS ON STUDY 

FIGURE 2 

Growth Curves for Male and Female Rats Administered Scopolamine Hydrobromide Trihydrate 

in Water by Gavage for 2 Years 
40 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE6 

Mean Body Weights and Survival of Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Weeks Vehicle Control 1 me/ke 5 melk 25 me/k 

on Av. Wt. No. of Av. Wt.Wt. (% of No. of Av. Wt.Wt. (96of No. of Av. Wt.Wt. (% of No. of 

Study (8) Survivors (g) controls) Survivors (g) controls) Survivors (g) controls) Survivors 

1 131 60 129 99 60 129 99 60 128 98 60 

2 157 60 157 100 60 157 100 60 153 97 60 

3 192 60 192 100 60 188 98 60 184 96 60 

4 222 60 22 1 100 60 217 98 60 212 95 60 

5 242 60 24 1 100 60 237 98 60 229 95 60 

6 257 60 255 99 60 248 96 60 242 94 60 

7 272 60 270 99 60 263 97 60 254 93 60 

8 286 60 285 100 60 276 97 60 265 93 60 

9 296 60 298 101 60 288 97 60 278 94 60 

10 308 60 307 99 60 298 97 60 287 93 60 

11 317 60 316 100 60 306 97 60 295 93 60 

12 325 60 324 100 60 317 98 60 304 93 60 

13 334 60 338 101 60 328 98 60 313 94 60 

17 356 60 358 101 60 347 98 60 329 93 60 

21 375 60 382 102 60 369 98 60 349 93 59 

25 400 60 403 101 60 392 98 60 367 92 58 

29 415 60 416 100 60 408 98 59 377 91 58 

33 425 60 433 102 60 416 98 59 388 92 58 

37 43 1 60 439 102 60 432 100 59 402 93 58 

41 441 59 445 101 60 437 99 59 404 92 58 

45 457 59 461 101 59 448 98 59 414 91 58 

49 462 59 467 101 59 457 99 59 419 91 58 

53 469 59 473 101 59 463 99 59 425 91 57 

57 475 59 479 101 59 469 99 59 429 90 57 

61 479 59 482 101 58 472 99 59 433 90 56 

65 479 57 488 102 57 477 100 59 437 91 56 

69a 483 46 492 102 47 487 101 48 442 92 46 

73 485 45 495 102 44 485 100 48 439 91 45 

77 482 43 495 103 44 48 1 100 48 438 91 44 

81 483 43 493 102 39 485 101 48 439 91 43 

85 479 41 485 101 37 474 99 45 429 90 40 

89 473 40 489 103 34 468 99 42 434 92 38 

93 468 38 478 102 32 448 96 36 424 91 38 

97 446 30 462 104 25 448 101 30 416 93 36 

101 431 26 449 104 20 43 8 102 25 422 98 29 

Mean for weeks 

1-13 257 256 100 250 97 242 94 

14-52 418 423 101 412 99 383 92 

53-101 472 482 102 469 99 43 1 91 

a Interimevaluation occurred atweek 66. 
Results 41 

TABLE7 

Mean Body Weights and Survival of Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Weeks Vehicle Control 1mg/k 5 mg/b 25 mdh 

on Av. Wt. No. of Av. Wt. Wt. (56 of No. of Av. Wt.Wt. (% of No. of Av. Wt.Wt. (56 of No. of 

Study (9) Survivors (8) controls) Survivors (8) controls) survivors (9) . controls) Survivors 

1 107 60 107 100 60 107 100 60 105 99 60 

2 124 60 125 101 60 122 98 60 122 98 60 

3 136 60 136 100 60 136 100 60 135 99 60 

4 148 60 149 101 60 148 100 60 145 99 60 

5 158 60 159 101 60 157 100 60 155 98 60 

6 162 60 161 100 60 161 99 60 157 97 60 

7 168 60 168 100 60 167 99 60 164 98 59 

8 174 60 173 100 60 170 98 60 1 67 96 59 

9 179 60 180 101 60 178 99 60 172 96 58 

10 184 60 186 101 60 184 100 60 178 97 58 

11 184 60 185 101 60 185 100 60 178 97 58 

12 190 60 190 100 60 188 99 60 181 95 58 

13 193 60 192 99 60 191 99 60 184 95 58 

17 201 60 200 100 60 1 97 98 60 192 96 58 

21 207 60 208 100 60 203 98 60 197 95 58 

25 215 60 216 101 60 214 100 60 203 94 58 

29 223 60 223 100 59 219 98 60 208 93 56 

33 229 60 226 99 43 226 99 60 212 93 56 

37 237 60 239 101 42 234 99 59 22 1 93 55 

41 245 60 243 99 42 239 98 59 226 92 55 

45 255 60 255 100 41 248 97 59 226 89 54 

49 266 60 I 264 99 41 259 97 58 240 90 51 

53 27 1 60 271 100 39 267 99 58 240 88 50 

57 282 59 277 98 36 275 97 57 25 1 89 46 

61 288 59 282 98 34 282 98 56 255 89 46 

65 295 58 288 98 34 291 99 56 264 90 46 

69a 306 46 298 98 31 300 98 44 267 87 35 

73 313 46 300 96 31 306 98 44 266 85 31 

77 318 46 306 96 29 311 98 44 271 85 30 

81 320 46 315 99 26 312 98 41 278 87 30 

85 328 43 308 94 23 316 96 41 280 85 28 

89 332 41 306 92 22 314 95 40 275 83 25 

93 335 40 32 1 96 17 315 94 35 274 82 24 

97 329 36 323 98 17 312 95 33 272 83 22 

101 335 34 325 97 17 314 94 29 270 81 22 

Mean for weeks 

1-13 1 62 162 100 161 99 157 97 

14-52 231 230 100 227 98 214 93 

53-101 312 302 97 301 96 266 85 

a Interimevaluation occurred atweek 66. Duetomortalityinvolvingthefloodingincidentatweek 30, no females receiving 1 mglkgwere 

included in the interim evaluation. 
42 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

Pathology and Statistical Analysis 

Thissectiondescribes thestatisticallysignificantor 

biologically noteworthy changes in the incidences of 

neoplasms and nonneoplastic lesions of the pituitary 

gland and incidences of mononuclear cell leukemia. 

Summaries of theincidencesofneoplasmsand 

nonneoplastic individual tumor lesions, animal 

diagnoses,statisticalanalysis of primary neoplasms 

that occurred with incidence of at least5 % in at least 

oneanimal group, and historicalincidences for the 

neoplasms mentioned inthis section are presented in 

Appendix A for male rats and AppendixB for female 

rats. 

Pituitary Gland: The incidences of adenomaofthe 

control, 19/49; 1 mg/kg, 17/49; 5 mg/kg,

13/50; 25 mg/kg, 10/50; females: 20/50, 13/60, 

14/50, 10150; Tables A3 and B3). The incidences of 

pituitaryglandparsdistalisadenomain25mg/kg 

males andallgroupsofdosedfemaleswere below 

those observed in historical control data from recent 

NTP 2-year gavage studies. In general, the incidences of hyperplasia werenotsignificantlydifferent 

from those in the control groups (TablesA5 and BS). 

MononuclearCellLeukemia: Theincidencesof 

mononuclearcellleukemiain 25 mg/kgmalesand 

femalesweresignificantlylowerthanthoseofthe 

control (Tables groups 8, A3,B3). andThe

incidence of mononuclear cellleukemia in females 

pituitary gland pars distalis decreased with increasingreceiving 25 mg/kgwaswellbelowthehistorical 

doseinbothmaleandfemalerats;however,this controlrangeobservedinrecent2-yeargavage 

trendwasonlysignificantinmales(males:vehicle studies (Tables 8 and B4b). 

Table 8 

Incidences of Mononuclear Cell Leukemia in Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Male 

Mononuclear Cell Leukemiaa 

Overall rateb 

Adjusted rate' 

Terminal rated 

First incidence (days) 

Life table teste 

Female 

Mononuclear Cell Leukemiaf 

Overall rate 

33/50 (66%) 21150 (42 46) 26/50 (52%) 24/50 (48%) 

86.0% 73.3% 66.5% 59.3% 

15/20 (75%) 8/14 (57%) 10/22 (45%) 12/28 (43%) 

443 533 463 504 

P=0.034N P=0.240N P=0.139N P=0.017N 

20150 (40%) 6/60 (10%) 13/50 (26%) 4/50 (8%) 

Adjusted rate 47.7% 26.9%13.9% 38.8% 

7/26 (27 R) 

555 

Terminal rate 

First incidence (days) 

13/34 (38%) 

374 

2/17 (12%) 2/22 (9%) 

392 519 

Life table test P=0.022N P=0.138N P=0.307N P=O.OllN 

Historical incidence for 2-year NTP water gavage studies with vehicle control groups (mean f standard deviation): 1731367 

(47.1%f 9.2%);range 34%-56% 

Number of animals with neoplasm per number of animals necropsied 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence in animals surviving until the end of the study 

In the control column are the P values associated with the trend test. In the dosed group columns are theP values correspondingto the 

painvise comparisons between the control and that dosed group. The life table test considers neoplasms in animals dying prior to the 

terminal kill as (directly or indirectly) the cause of death. A negative trend or lower incidence in a dosed group is indicated by N. 

Historicalincidence: 99/368 (26.9% f 7.6%);range 16964% 
Results 43 

MICE Theabsoluteorganweightsofdosedmaleswere 

similar to those of the control group (TableF4). The 16-DAY STUDY relative liver weights of males receiving1,800 mg/kg Onemaleand two femalesreceiving 1,800 mg/kg and of females in all dosed groups were significantly and one female receiving 150 mg/kg died during the greater than those of the control group. There were study(Table 9). Thefinalmeanbodyweightsand no significant incidences of treatment-related gross or bodyweightgainsofdosedmiceweresimilar to microscopic lesions. those of the control groups. Clinical findings related 

toscopolamineadministrationincludedbilateral Based on mortalityamong 1,800 mg/kgmalesand pupillarydilationandsquintinginalldosedmales females, doses selected for the 14-week studywere and females. 15, 45, 135, 400 and 1,200 mg/kg. 

TABLE9 

Survival and Body Weights of Mice in the 16-Day Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Mean Body Weightb (g) Final Weight 

Initial Survivala Dose Final Changeto Controls Relative

(mgki!) (W 

Male 

0 515 20.2 f 0.5 22.4 f 0.2 2.2 f 0.5 

150 515 20.8 f 0.4 22.6 f 0.5 1.8 f 0.6 101 

250 515 20.4 f 0.8 21.6 f 0.6 1.2 f 0.5 96 

450 515 20.6 f 0.5 22.6 f 0.7 2.0 f 0.6 101 

900 515 20.6 f 0.2 22.0 f 0.5 1.4 f 0.4 98 

1,800 415' 21.2 f 0.6 22.8 f 0.9 1.3 f 0.3 102 

Female 

0 515 18.2 f 0.4 19.8 f 0.6 1.6 f 0.5 

150 415' 18.6 f 0.2 19.0 f 0.4 0.3 f 0.3 96 

250 515 18.0 f 0.3 19.2 f 0.2 1.2 f 0.4 97 

450 515 18.0 f 0.6 18.8 f 0.4 0.8 f 0.4 95 

900 SI5 18.0 f 0.6 18.8 f 0.5 0.8 f 0.5 95 

1,800 3d 18.2 f 0.5 18.3 f 0.7 0.0 f 0.0 93 

a Number of animals survivinglnumber initially in group. Subsequent calculations are based on animals surviving to theendofthestudy. 

Weights and weight changes are given as mean f standard error. Differences from the control are not significant by Williams' or 

Dunnett's test. ' Dayofdeath: 4 

Day of death: 4.4 
44 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

Iq-WEEK STUDY and/or rate of release from the bone marrow, shifts 

Onemalereceiving135mg/kgandtwomalesand in the distributionof neutrophils between the marginonefemalereceiving 1,200 mg/kg diedduringthe al and circulating vascular pools, or increases in the 

study (Table 10). The final meanbodyweightsand intravascular neutrophil life span. 

mean body weight gains of all dosed male groups andThe estrous cycle lengthof 1,200 mg/kg females was females receiving 45 mg/kg and above were significantlylowerthanthose of thecontrolgroups. 

Clinicalobservationsincludedbilateralpupillary 

dilation, hyperactivity, and hypoactivity. 

The hematology data for mice in the 14-week study 

arelistedinTable G3. A minimaltomildmature 

neutrophilia,similar to that whichoccurredinthe 

14-week rat study, occurred in male mice receiving 

45 mg/kg or higher. As with the rat study, there was 

no microscopic evidence of inflammation that could 

account for the neutrophilia. Mechanisms that could 

beconsideredinclude:alterationsingranulopoiesis 

TABLZ10 

significantlygreaterthanthatinthecontrolgroup 

(Table H2). Sperm morphology parameters in dosed 

males were similar to those in the control group. 

Therewerenosignificantdifferencesinorgan 

weights (Table F5) or incidences of treatment-related 

gross or microscopic lesions. 

DoseSelectionRationale: Based on lowersurvival 

in 1,200 mg/kg mice and lower mean body weights 

of mice exposed to 45, 135, 400, and 1,200 mg/kg , 

doses selected for use in the 2-year study were 1, 5, 

and 25 mg/kg. 

Survival and Body Weights of Mice in the 14-Week Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

MeanBodyWeightb (g) 

Initial Survivala Dose Final 

(mgkg) 

Male 

0 10110 23.8 f 0.4 32.7 f. 1.0 

15 10110 24.6 f 0.3 29.1 f 0.4** 

45 10110 24.2 f 0.3 28.8 f 0.6** 

135 91 10' 24.3 f 0.4 29.3 f 0.6** 

400 10110 24.4 f 0.4 28.7 f 0.8** 

1,200 81lOd 24.1 f 0.3 28.9 f 0.6** 

Female 

0 1011019.2 f 0.2 - 28.0 f 0.5 

15 1011018.9 f 0.5 26.9 f 0.4 

45 101109.7 f 0.226.5 f 0.3*6.8 

135 1011019.5 f 0.5 26.1 f 0.4**6.6 

400 1011019.4 f 0.326.2 f OS** 

1,200 9/10'19.2 f 0.225.5 f 0.4**6.4 

ChangeRelative 

(%I 

to Controls 

Final Weight 

8.9 f 0.8 

4.5 f OS** 

4.6 f 0.6** 

4.9 f 0.6** 

4.3 f 0.8** 

4.2 f 0.6** 88 

88 

89 

88 

89 

f 0.4** 

6.8 f 0.3** 

f 0.6** 

f 0.4** 

8.0 f 0.4 

8.8 f 0.5 

91 

93 

93 

95 

96 

* Significantly different (P SO.05) from thecontrolgroupbyWilliams'orDunnett's test 

** PSO.01 

a Number of animalssurvivinglnumberinitiallyingroup.Subsequentcalculationsarebased on animals surviving to theend of the study. 

Weights and weight changes are given as mean f standarderror.

' Week of death: 10 

Week of death: 10, 12 

e Week of death: 1 
Results 45 

%YEAR STUDY control groups throughout the study (Figure 4; 

Tables 12 and 13). The mean body weights of males Survival 

andfemalesreceiving 5 mg/kg were slightly lower Estimatesof2-year survival probabilities for male thanthoseof the control groups. The mean body and female mice are shown inTable 11 and inthe weightsofmalesandfemalesreceiving 25 mg/kg Kaplan-Meier survival curves (Figure 3). Survival of werelowerthan those of the control groups after dosedmaleandfemaleswas similar to thatofthe week 13. The final mean body weights of males and controls. femalesreceiving 25 mg/kg were 19% (males) and 

16% (females) lower than those of the control 

Body Weights, Clinical Findings, groups. Clinical findings included bilateral pupillary 

and Ophthalmic Examination Findings dilation in all dosed maleand female groups. 

The meanbodyweightsofmalesandfemales Ophthalmicexaminationsresultedin no significant 

receiving 1 mg/kg weresimilar to thoseof the findings. 

TABLE11 

Survival of Mice in the 2-Year Gavage Studyof Scopolamine Hydrobromide "hydrate 

Male 

in initially Animals 70 70 70 

15-Month interim evaluationasb 

Accidental deathsa 

Moribund 

Natural deaths 

Animals surviving to study termination 

Percent probability of survival at the end of study' 

Mean survival (days)d 

20 

0 

4 

6 

40 

81 

632 635 

79 

39 

4 

7 

0 

20 

614 

82 

39 

5 

4 

2 

20 

622 

83 

39 

2 

7 

2 

20 

P=0.934N survival analysise P=O.992 P= 1.000N P= 1.000N 

Female 

in initially Animals 70 70 70 

15-Month interim evaluationasb 19 20 20 19 

Accidental deathsa 2 0 1 0 

Moribund 9 11 7 7 

Natural deaths 7 3 5 6 

Animals surviving to study termination 33 36 37 38 

Percent probability of survival at the end of study 67 73 77 76 

Mean survival (days) 617 624 614 624 

ysis Survival P=0.835N P=0.571N P=O.656 

a Censoredfromsurvivalanalyses 

Includes 10 animals per group that received ophthalmic examinations only 

Kaplan-Meier determinations 

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

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

1972) with the controls are in the dosed columns. A negative trend or lower mortality in a dose group is indicated by N. 
46 

~~ 

Scopolamine Hydrobromide Trihydrate,NTP TR 445 

I .o 

-I 0.9 a

L 

s

3 

v) 

L 

......................................... 0.8 r:

d 

0 

m 

a 

m 

s 

a. 0.7 

0 1 MG/KG 

0.6 

15 

I .c 

.-................... -4 0.9 a

2

> 

e

3 

v) 

L 

0 ........................... 

-i= Os8 

-t 

m 

a 

m 

s 

n 0.7 

0 1 YG/UG

A SHG/KC 

0.6 I 

I5 

-

.... 

.... 

I............................................................................... : 4 

............. ................. ....... .... 

- so 45 60 75 90 1c 

WEEKS ON STUDY 

... ... ..... 

......... ..... 

.... .......... ... .... ....... ................................ ..... ~ yi I I I 1 I 

30 45 60 75 90 1 5 

WEEKS ON STUDY 

FIGURE 3 

Kaplan-Meier Survival Curves for Male and Female Mice Administered 

Scopolamine Hydrobromide Trihydrate in Water by Gavage for 2 Years 
47 Results 

.................. ^..... .................................................................................................... ."................... 

................. j......................................... 

.................. .: ................. ....... ................. .................... .................. 

I . :A ! 00,~0~0~0~ ................... :.............o...& .................... ;....a. ..a,. 13. .p.................. :. ................... .$.............o.. . 

i. 0 : -A i 0'0 : - n 

-

pia:u ............. a..4............... i.................... .i..................... j..................... :....................

4; 00 : 

$io 

R.j. ............................................................................................................................. 

............... ..................... ;.................... 1..................... ;.................. MALE MICE ..... 

.......................... .................................. ..................................... 

1 I I I I I 

15 30 45 60 ' 75 90 1 

WEEKS ON STUDY 

FIGURE 4 

Growth Curves for Male and Female Mice Administered Scopolamine Hydrobromide Trihydrate 

in Water by Gavage for 2 Years 
48 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE 12 

Mean Body Weights and Survival of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Vehicle WeeksControl 1mdke 5 melk 25 mdke

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

Study 0 Survivors (9) controls) Survivors (9) controls) Survivors (g) controls) Survivors 

1 24.3 70 24.1 99 70 24.2 100 70 24.1 99 70 

2 26.1 70 25.4 97 70 25.5 98 70 25.4 97 70 

3 26.6 70 26.1 98 70 26.2 99 70 25.7 97 70 

4 28.0 70 27.9 100 70 27.5 98 70 27.0 96 70 

5 29.2 70 28.8 99 70 28.6 98 70 27.8 95 70 

6 30.2 70 29.5 98 70 29.1 96 70 27.8 92 70 

7 30.9 70 30.3 98 70 29.5 96 70 28.2 91 70 

8 31.4 70 30.8 98 70 30.4 97 69 28.5 91 70 

9 31.9 70 31.1 98 70 30.3 95 69 28.7 90 70 

10 32.5 70 31.8 98 70 30.9 95 69 29.2 90 70 

11 33.4 70 32.5 97 70 31.3 94 69 30.1 90 70 

12 33.7 70 32.9 98 70 32.0 95 69 30.5 91 70 

13 34.5 70 33.2 96 70 32.4 94 69 30.5 88 70 

17 38.0 70 36.5 96 70 34.9 92 69 31.9 84 70 

21 40.9 70 39.5 97 70 37.2 91 68 33.4 82 70 

25 42.1 70 40.7 97 70 38.5 91 68 34.0 81 70 

29 43.8 70 43.O 98 70 40.3 92 68 34.9 80 70 

33 45.5 70 44.8 99 70 41.6 91 68 35.6 78 70 

37 47.9 70 46.0 96 70 43.9 92 68 37.6 79 70 

41 47.8 70 47.2 99 70 45.1 94 68 38.3 80 70 

45 49.7 69 47.4 95 70 46.3 93 68 38.9 78 69 

49 49.7 69 48.7 98 70 46.5 94 68 39.7 80 69 

53 49.7 69 48.7 98 70 47.0 95 68 39.8 80 69 

57 49.8 69 49.3 99 70 7.2 95 67 39.1 79 69 

61 50.5 69 49.6 98 69 47.8 95 67 39.8 79 67 

65 50.9 67 49.7 98 69 48.1 95 67 40.8 80 66 

69a 51.7 48 49.6 96 49 48.3 93 47 41.6 81 46 

73 52.0 48 49.2 95 49 47.8 92 47 40.8 79 46 

77 52.4 48 49.1 94 49 48.4 92 46 41.4 79 46 

81 51.9 41 48.8 94 48 47.4 91 45 41.1 79 46 

85 51.4 47 48.2 94 48 46.5 91 44 41.3 80 45 

89 51.8 47 48.4 93 46 47.6 92 42 41.3 80 44 

93 51.2 46 48.4 95 46 46.9 92 42 41.5 81 42 

97 49.8 46 48.6 98 44 46.7 94 42 41.O 82 42 

101 49.7 42 47.6 96 42 45.5 92 40 40.1 81 40 

Mean for weeks 

1-13 30.2 29.6 98 29.1 96 28.0 93 

14-52 45.O 43.8 97 41.6 92 36.0 80 

48.9 96 47.3 93 40.7 80 

a Interim evaluation occurredatweek 66. Includes 10 animalspergroupthatreceivedophthalmicexaminations only. 

53-10151.0 
Results 49 

TABLE13 

Mean Body Weights and Survival of Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Weeks Vehicle Control 1mglkg 5 mglkg 25 rnelke 

on Av.Wt. No. ofAv.Wt.Wt. (aof No. of Av.Wt.Wt. (96of No. of Av.Wt.Wt. (kof No. of 

Study (fd Survivors (8) controls) Survivors (9) controls) Survivors (g) controls) sunivors

1 19.8 70 19.8 100 70 19.6 99 70 19.7 100 70 

2 21.4 69 21.7 101 70 21.3 100 70 20.9 98 70 

3 22.6 69 23.1 102 70 22.9 101 70 22.4 99 70 

4 23.9 69 24.1 101 70 23.8 100 70 23.2 97 70 

5 25.0 69 25.1 100 70 24.7 99 70 24.1 96 70 

6 25.6 69 25.8 101 70 25.1 98 70b 24.4 95 70 

7 26.6 69 26.4 99 70 26.2 99 70 25.3 95 70 

8 28.2 69 28.0 99 70 27.4 97 70 26.2 93 70 

9 28.7 69 28.4 99 70 27.5 96 70 26.3 92 70 

10 29.6 69 28.9 98 70 27.8 94 70 26.5 90 70 

11 30.1 69 29.8 99 70 28.9 96 70 27.1 90 70 

12 30.7 69 30.6 100 70 29.4 96 70 27.6 90 70 

13 31.2 69 30.9 99 70 29.6 95 70 27.6 89 70 

17 34.8 69 33.9 97 70 32.0 92 70 29.3 84 70 

21 37.7 69 36.8 98 70 35.4 94 70 31.8 84 70 

25 38.7 69 37.9 98 70 35.6 92 70 32.1 83 70 

29 41.O 69 40.3 98 70 37.9 92 70 33.6 82 70 

33 43.5 69 41.8 96 70 39.2 90 70 35.1 81 70 

37 45.8 69 43.9 96 70 41.O 90 69 36.2 78 70 

41 47.0 69 45.4 97 70 42.4 90 69 37.9 81 70 

45 49.5 69 47.6 96 70 44.5 90 68 38.5 78 70 

49 51.0 69 49.6 97 70 46.1 90 68 39.1 77 70 

53 51.4 69 50.4 98 70 46.4 90 67 39.7 77 69 

57 52.0 68 51.1 98 70 46.9 90 66 39.5 76 69 

61 53.0 68 51.6 97 69 47.8 90 66 40.0 76 68 

65 54.0 68 52.4 97 68 48.9 91 66 41.1 76 67 

69' 54.7 49 53.7 98 48 49.9 91 46 42.3 77 47 

73 56.0 48 54.5 97 47 50.0 89 45 42.0 75 47 

77 55.3 47 54.6 99 46 49.6 90 45 42.0 76 47 

81 53.9 47 53.9 100 45 48.2 89 45 41.4 77 45 

85 52.9 46 52.9 100 44 48.0 91 44 41.6 79 45 

89 53.3 40 53.7 101 43 48.6 91 42 42.3 79 44 

93 52.4 40 53.2 102 42 47.7 91 41 42.4 81 43 

97 50.2 39 51.9 103 41 47.2 94 40 41.1 82 41 

101 47.9 38 50.7 106 38 45.4 95 38 40.3 84 38 

Mean for weeks 

1-13 26.4 26.4 100 25.7 91 24.7 94 

14-52 43.2 41.9 97 39.3 91 34.8 81 

52.7 100 48.0 91 41.2 78 

a Interimevaluationoccurred at week 66. Includes 10 animals per group thatreceivedophthalmicexaminations only. 

Number of animals weighed for this week was less than thenumber of animals surviving 

53-10152.8 
50 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

Hematology

Thehematologydata for mice atthe15-month 

interimevaluationinthe2-yearstudyarelistedin 

TableG4.Hematocritlevel,hemoglobinconcentration, and erythrocyte count in25 mg/kg female mice 

were slightly lower than those in the control group. 

This result would be consistent with development of 

a minimal normocytic, normochromic nonresponsive 

anemia.Theanemiawouldbeconsistentwiththe 

lowerbodyweightsexhibitedbytheseanimalsand 

wouldbepresumedtoberelatedto a decreased 

nutritionalstatus.Themousehematologyresults 

differ from those which occurred in the rat study. It 

incidences for the neoplasmsmentionedinthis 

section are presented in Appendix C for male mice 

and Appendix D for female mice. 

Liver: Theincidences of hepatocellular adenomain 

5 mg/kg males and 25 mg/kg males and females were 

significantly lower than those of the control groups 

(Tables14,C3,andD3).Theincidence of hepatocellular carcinoma in 1 mg/kg males was significantly 

greater than that in the control group. The combined 

incidencesofhepatocellularneoplasmsfollowed a 

significantnegativetrendinmalesandfemales. A 

hepatoblastomaoccurred in one 1 mg/kg male. The 

should be noted that all erythron changes, in both thecombined incidences of hepatocellular neoplasms in 

rat and mouse studies, were minimal and that hemoconcentrationrelatedtodehydrationcouldmask a 

minimal anemia. 

Pathology and Statistical Analysis 

This sectiondescribesthestatistically significantor 

biologically noteworthy changes in the incidencesof 

neoplasms and/or nonneoplastic lesions of theliver 

andotherorgans.Summaries oftheincidencesof 

neoplasmsandnonneoplasticlesions,individual 

animaltumordiagnoses, analysis statistical of 

primary neoplasms that occurred with incidenceof at 

least 5% in at least one animal group, and historical 

control males and females, 1 mg/kg male mice, and 

all female dosed groups exceeded the range observed 

incontrolmiceinhistoricalNTP2-yeargavage 

studies(Tables 14, C4a, andD4a).Thecombined 

incidencesofneoplasmsin 5 and25mg/kgmales 

were within the NTP historical control range. 

The incidences of clear cell foci and eosinophilic foci 

in dosed male groups were significantly lower than 

those of the control group, and decreasedwith 

increasingdose(Tables14and C5). Theincidence 

of eosinophilicfocusin 25 mg/kgfemaleswas 

significantlylowerthanthatinthecontrolgroup 

(Tables 14 and DS). 
Results 51 

TABLE14 

Incidences of Neoplasms and Nonneoplastic Lesionsof the Liver of Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate 

Vehicle Control 

Male 

15-Month Interim Evaluation 

Number Examined Microscopically 10 

Clear Cell Focusa 1 

Eosinophilic Focus 1 

Mixed Cell Focus 1 

Hepatocellular Carcinoma 

Hepatocellular Adenoma 

0 

1 

2-Year Study 

Number Examined Microscopically 50 

Basophilic Focus 3 

Clear Cell Focus 12 

Eosinophilic Focus 21 

Mixed Cell Focus 5 

Hepatocellular Adenoma 26 

Hepatocellular Carcinoma 6 

Hepatocellular Adenoma or Carcinomab 

Overall rate' 30/50 (60%) 

Adjusted rated 65.2% 

Terminal ratee 24/40 (60%) 

First incidence (days) 680 

Logistic regression testf P=0.001N 

(continued) 

1 mgh 

10 

1 

2 

0 

2 

0 

50 

4 

4* 

12* 

7 

22 

IS* 

33/50 (66%) 

67.3% 

23/39 (59%) 

405 

P=O.348 

5 mgkg 

10 

1 

1 

2 

1 

0 

50 

3 

2** 

7** 

7 

9** 

6 

14/50 (28%) 

32.5% 

10139 (26%) 

594 

P=0.002N 

25 mgkg 

10 

0 

0 

0 

0 

1 

50 

0 

0** 

2** 

1 

8** 

7 

15/50 (30%) 

35.3% 

12/39 (3146) 

587 

P=0.004N 
52 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE14 

Incidences of Neoplasms and Nonneoplastic Lesionsof the Liver of Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Female 

15-MonthInterim Evaluation 

Number Examined Microscopically 

Basophilic Focus 

Mixed Cell Focus 

Hepatocellular Adenoma 

2-Year Study 

Number Examined Microscopically 

Basophilic Focus 

Clear cell Focus 

Eosinophilic Focus 

MixedcellFocus . 

Hepatocellular Adenoma 

Hepatocellular Carcinoma 

Hepatocellular Adenomaor Carcinomag 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Logistic regression test 

* 

Vehicle Control 1 mgk 25 mgkg 

10 10 10 10 

0 0 1 0 

1 1 0 0 

1 0 0 0 

51 50 50 51 

1 2 3 3 

0 1 0 0 

17 10 13 9* 

6 6 3 3 

15 18 9 6* 

8 6 8 4 

22/51 (43%) 21/50 (42 %) 16/50 (32%) 9/51 (18%) 

57.1% 55.0% 37.0% 23.1% 

17/33 (52%) 19/36 (53 %) 10137 (27%) 8/38 (21 k) 

594 676 578 694 

P=0.002N P=0.466N P=0.165N P=0.003N 

Significantly different(P50.05) from the control group by the Fisher exact test (15-month interim evaluation) or the logistic regression 

test (2-year study) ** P50.01 a Number of animals with lesion 

b Historical incidence for 2-year NTP water gavage studies with vehicle control groups (mean f standard deviation): 74/315 

(23.5%f 7.2%);range 14%-36% 

Number of animals with neoplasm per number of animals with liver examined microscopically 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence in animals surviving until the end of the study 

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

painvise comparisons between the control and that dosed group. The logistic regression test regards neoplasms occurring in animals 

prior to the terminal kill as nonfatal. A negative trend or lower incidence in a dosed group is indicated by N. 

Historicalincidence: 21/315 (6.7% f 4.2%); range 2%-12% 
5 

Results 53 

incidence alveolar hyperplasia 

eously occurring nonneoplastic lesions were signifi- 25 mg/kg males was significantly lower than that of 

cantlylowerindosedmaleandfemalemicethan thecontrolgroup.Theincidence of bonemarrow 

thoseinthecontrolgroupsandusuallydecreased myelofibrosis in 25 mg/kg females was significantly 

withincreasingdose(Tables 15, C5, and D5). At lowerthanthatinthecontrolgroup.Incidencesof 

15 months,theincidence ofkidneynephropathyin hyperplasia of thepituitaryglandparsdistalisin 

25 mg/kg males was significantly lower than that in and 25 mg/kg females and cystic hyperplasia of the 

thecontrolgroup. At 2 years,theincidences of uterusandhematopoieticcellproliferationinthe 

kidney nephropathy in 25 mg/kg males and females spleenin 25 mg/kgfemaleswerealsosignificantly 

weresignificantlylowerthantheincidencesinthe lower than those in the control group. The decreased 

control groups. Compared to the control group, the incidencesofthesespontaneouslesionsweremost 

incidences of hyperplasia of the pancreatic islets in5 likely a result of lower body weights in dosed versus 

and 25 mg/kgmalesweresignificantlylower.The control animals. 

Other Organs: Theincidences ofmany spontan- of epithelial in 

TABLE15 

Incidences of Selected Nonneoplastic Lesions in Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

Vehicle Control 1 mglkg 5 mgk 25 mglkg 

Male 

15-MonthInterimEvaluation 

Kidneya 10 10 10 10 

Nephropathyb 10 (1.2y 9 (1.0) 10 (1.0) 6* (1.0) 

Pancreatic Islets 10 10 10 10 

Hyperplasia 3 (1.3) 7 (1.4) 4 (1.5) 0 

2-Year Study 

Kidney 50 50 50 50 

Nephropathy 48 (1.0) 46 (1.3) 42 (1.1) 37**(1.1) 

Lung 50 50 50 50 

Alveolar Epithelial Hyperplasia 8 (1.5) 3 (2.0) 2 (1.0) 1* (1.0) 

Pancreatic Islets 50 50 50 50 

Hyperplasia 29 (1.6) 23 (1.4) 8**(1.3) 2** (2.5) 

(continued) 
54 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE 15 

Incidences of Selected Nonneoplastic Lesions in Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle Control 1 mg& 5 mg& 25 mgkg 

Female 

15-Month Interim Evaluation 

Bone marrow 10 10 10 10 

Myelofibrosis 1 (1.0) (1.5) 2 0 0 

Kidney 10 10 10 10 

Nephropathy 5 (1.0) (1.0) 3 1 (1.0) (1.0) 2

Pituitary Gland (Pars Distalis) 10 10 10 10 

Hyperplasia 3 (1.0) 0 0 (1.0) 1

UteNS 

Cystic Hyperplasia 

2-Year Study 

Bone Marrow 

Myelofibrosis 

Kidney 

Nephropathy 

Pituitary Gland (Pars Distalis) 

Hyperplasia 

Spleen 

Hematopoietic Cell Proliferation 

10 

5 

51 

(1.2) 2

51 

(1.0) 23

50 

24 

51 

(1.9) 17

(2.2) 

(1.8) 

10 

8 

50 

21 

50 

21 

47 

(1.5) 15

50 

(2.3) 11

(1.4) 

(1.1) 

(1.0) 

10 

4 (2.0) 

50 

(1.3) 15

50 

(1.0) 26

47 

11**(1.3) 

50 

(1.9) 17

10 

(1.5) 4

51 

13*(1.2) 

51 

10**(1.1) 

46 

13*(1.8) 

51 

7* (2.3) 

uterus 50 51 50 51

Cystic Hyperplasia 38 (2.3) 29* (2.1) 30 (1.9) 23**(1.5) 

* Significantly different(PSO.05) from the control group by the Fisher exact test (15-month interim evaluation) or the logistic regression 

test (2-year study)

** PSO.01 

a Numberofmicewith orgadtissue examined microscopically 

Number of mice with lesion 

Average severity grade of lesions in affected animals(1=minimal; 2=mild; 3=moderate; 4=marked) 
Results 55 

GENETICTOXICOLOGY 

Scopolamine hydrobromide trihydrate (100 to 

10,000 pglplate) did not induce mutations in Salmonella typhimurium strains TA97, TA98, TA100, 

TA1535, or TA1537, with or withoutAroclor 

1254-inducedmaleSprague-Dawleyrat or Syrian 

hamster liver S9 (Table El). In cytogenetic tests 

with cultured Chinese hamster ovary cells, no convincing induction of sister chromatid exchanges was 

notedwithscopolamine hydrobromide trihydrate 

doses up to 500 pg/mL without S9 or 5,000 pg/mL 

with S9 (Table E2). Results from the first two trials 

conducted with S9, which appeared to clearly demonstrate a significantincreasein sister chromatid 

exchanges, were called into question by the observation of a pH shift in the culture medium produced by 

the high doses (2,000 pg/mL and higher) of scopolamine hydrobromide trihydrate, which coincided with 

theincreasesin sister chromatid exchanges.Therefore, a third trial was conducted, in which the pH of 

the culture medium was adjusted with N-(2-hydroxymethyl)piperazine-N’-(2-ethanesulfonic acid) 

(HEPES) buffer. Resultsof this third trial were 

negativeandthe overall assay results were also 

considered to be negative. The increases in sister 

chromatidexchangesnotedinthe presence of S9 

were attributed to the alteration in pH produced by 

highconcentrationsofscopolamine hydrobromide 

trihydrate. No induction of chromosomal aberrations 

was observed in cultured Chinese hamster ovary cells 

treatedwithscopolamine hydrobromide trihydrate 

without S9, but with S9, even in the presence of 

HEPES buffer to maintain optimum pH, increases in 

the percentage of cells with aberrations were noted in 

eachof two trials, at the highest dose tested 

(5,000 pg/mL) (Table E3). 

Despite the evidence for induction of chromosomal 

damageinculturedChinesehamster ovary cells in 

vitro, no increase in the frequency of micronucleated 

normochromatic erythrocytes was noted in peripheral 

blood samples obtained from male and female mice 

at the end of the 14-week gavage studies of scopolamine hydrobromide trihydrate (Table E4). 
56 Scopolamine Hydrobromide Trihydrate,NTP TR 445 
57 

DISCUSSION AND CONCLUSIONS 

Scopolamine hydrobromide trihydrate, a white 

crystalline or granular powder, is a solanaceous 

alkaloid derived from the Solanaceae family (Brown, 

1990). Its major use is in transdermal patches for the 

treatmentofmotion sickness. Scopolaminehydrobromide is an anticholinergic drug which is used for 

thetreatmentof various ailments includingacute 

mania, diarrhea, gastric and duodenal ulcers, gastrointestinal spasm, excessive salivationandsweating, 

.and infantile cerebral palsy. It is alsousedas a 

mydriaticand agentas cycloplegicand 

over-the-counter sleeping aid. 

Scopolamine hydrobromide trihydrate was nominated 

to the NTP for toxicology and carcinogenicity testing 

by the National Cancer Institute because of considerablehuman exposure resulting from its use as a 

prescription or over-the-counter drug and as a representative chemical from a class of alkaloids. Scopolamine hydrobromide trihydrate is a suspect carcinogen because it contains .an aliphatic epoxide moiety, 

which may act as a biologicalalkylatingagent. 

Toxicologyandcarcinogenicitystudieswereconductedby administering scopolamine hydrobromide 

trihydrate in distilled waterbygavageonce daily, 

5 days per week, for 16 days, 14 weeks, or 2 years 

to male and female F344/N rats and B6C3F, mice. 

The gavage route of exposure was chosenbecause 

this route mimics oral exposure in humans,and 

because higher doses can be administered t4 the 

animal than can be achieved by the dosed feed route 

of administration. 

The results of the 16-day and 14-week studies in rats 

suggest that male rats are less tolerant to scopolamine 

hydrobromide trihydrate than females. In the 16-day 

rat study, body weight depression was observedin 

males administered 300 mg/kg or greater; no significant body weight depression occurred indosed 

females. In the 14-week study, final meanbody 

weights of dosedmalesranged from 1 1 % to 19% 

less than the final mean body weight of the control 

group. In dosed female rats, final meanbody 

weights ranged from 3 % to 7% lower than the final 

meanbody weightof the control group. In the 

14-week study, survival of female rats was also 

higherthanthatofmales. Early deaths of dosed 

malesandfemaleswerecausedbyesophageal 

obstructionand concurrent tracheal obstruction, as 

indicated by the accumulation of feed at these sites. 

This obstruction was considered to be related to the 

inhibitory effect Qf scopolamine hydrobromide 

trihydrate on salivary gland secretions and on esophageal smooth muscle contractions (i.e. swallowing). 

In mice, no clear effects wereobserved in body 

weights or survival of dosed male or female animals 

duringthe16-day or the 14-week studies. The 

differences from controls of weights body in

1,200 mg/kg male and femalemice in the 14-week 

studyweresimilar (male: 12%; female: 9%). 

Similarnumbers of chemical-related deaths were 

observed in males and females receiving 1,800 mg/kg 

scopolamine hydrobromide trihydrate in the 16-day 

studyandinthosereceiving 1,200 mg/kg in the 

14-week study. 

The low absolute weights of various organs observed 

in rats and mice receiving scopolaminehydrobromide 

trihydrate in the 14-week studies were considered to 

be related to the body weight depressions observed in 

thesedosed groups because therelativeweightsof 

these organs in dosed rats and mice were not significantly different from those ofthe controls. The 

bilateral pupillary dilation observed in the 16-day and 

14-week studies in dosed rats and mice was a manifestation of the pharmacologic effect of scopolamine 

hydrobromide trihydrate. 

The increases inhematocritvaluesand hemoglobin 

concentrations observed in dosed rats in the 14-week 

studyweremildandcouldhavebeen the result of 

dehydration, while the increases in segmentedneutrophil counts observed in dosed rats could have been 

due to stress. Thesehematologic differences were 

probably secondary to the pharmacologic' effects of 

this chemical. 
58 

No significant chemical-related lesions were observed 

in rats or mice in the 16-dayandthe14-week 

studies. 

Based on the mortality and lower body weights 

observed in the 14-week studies, the doses of scopolamine hydrobromide trihydrate selected for the 

2-year studies were 1, 5, and 25 mg/kg for rats and 

mice. 

In the 2-year rat study, survival of 1 and 25 mg/kg 

females was lower than that of controls. Survival of 

male and female mice receiving scopolamine hydrobromide trihydrate was not significantly different 

from that of the control groups. Mean body weights 

of 25 mg/kg male andfemale rats and miceand 

5 mg/kg male and female mice werelower than those 

of the controls throughout the studies. Based on the 

lower body weightsand poor survival at higher 

doses, the doses used in the 2-year studies were 

considered to be sufficiently high for determining the 

potential carcinogenicity of scopolamine hydrobromide trihydrate. 

The therapeutic oral dose of scopolamine for humans 

is 0.6 mg (Goodman and Gilmn3, 1985).

Compared to the peak scopolamine plasma concentration observed in humansreceiving an oral dose of 

0.4 mg, the plasma concentrations of scopolamine 

hydrobromide trihydrate in rats from the 5 mg/kg 

group were 12 times greater and inthe 25 mg/kg 

groups the concentrations were 24 to 56 times 

greater. 

No increased incidences of neoplasms that could be 

attributedscopolamine hydrobromide trihydrate 

administration were observed in male or female rats. 

Dose-relatednegative trends in theincidencesof 

pituitary gland and cell adenomamononuclear 

leukemia were observed in maleandfemale rats. 

The incidences of these neoplasms in the 25 mg/kg 

groups were significantly lower than thoseinthe 

control groups (pituitary gland adenoma:vehicle 

control, 19/49; 25 mg/kg, 10150[males]; 20/50, 

10/50[females]; mononuclear cell leukemia: 33/50, 

24/50 [males]; 20/50, 4/50 [females]). 

The influence of bodyweight on the incidence of 

these hasreported neoplasmsbeen by other

Scopolamine Hydrobromide Trihydrate,NTP TR 445 

researchers. Seilkop (1995) found a positive relationshipbetweenbodyweightand the ' incidence of 

pituitary gland neoplasms in male and female F344 

rats. Bodyweight reduction, as a result of dietary 

restriction, also hasbeen shown to decrease the 

incidence of mononuclear cell leukemia in F344 rats 

(Masoro, 1993). The decreased incidence of pituitary neoplasms may be associated with lower body 

weightgains. However, lower body weight cannot 

totally account for the decreased incidence of leukemia observed in female rats. 

In the 2-year mouse study, no increased incidencesof 

neoplasms could be attributed to scopolamine hydrobromide trihydrate administration in males or females. There was a dose-related negative trend in 

theincidencesofhepatocellularneoplasms in both 

male and female mice. 

Liver tumor incidences in male and female B6C3Fl 

mice havebeen demonstrated to be correlated with 

bodyweight.Using NTP historical control data, 

Seilkop (1995) derived a logistic regression model for 

predicting liver tumor incidences based on survival 

and body weight at 1 year. Application of this model 

to the scopolamine hydrobromide trihydrate data is 

summarized in Table 16. 

With the exception of 5 mg/kg malemice,the 

observed liver neoplasm incidences are very similar 

to what would be expected in control animals of 

equivalentbodyweightand survival. Thus, the 

decreased incidences of liver neoplasms in male and 

female mice are primarily a reflection of the reduced 

bodyweightinthedosed groups. In addition, the 

meanbodyweightsof the control animals in this 

study were 25 % to 30% higher than those of control 

animals in previous water gavage studies in the NTP 

historical control database. This observation may 

.help explain therelatively high liver neoplasm 

incidences in control animals from this study compared with the NTP historical rates reported in this 

study. 

No increased incidences of nonneoplastic lesions in 

male or female mice could be attributed to scopolamine hydrobromide trihydrate administration; 

however, there were dose-related negative trends in 

the incidences of clear cell and eosinophilic foci of 
Discussion and Conclusions 59 

TABLE 16 

Analysis of Liver Neoplasm Incidences Using the Seilkop (1995) Logistic Regression Model 

Male 

na 

Mean l-Year Body Weights (9) 

Liver Neoplasm Incidence 

Observed 

Predicted 

Female 

n 

Mean 1-Year Body Weights (8) 

Liver Neoplasm Incidence 

Observed 

Predicted 

VehicleControl 1 mgkg 5 mgkg 25 mwg 

50 50 48 49 

50.0 48.6 46.7 39.9 

30 (60%) 

65% 

33 (66%)

61 96 

14 (29%) 

54% 

15 (31X)

34% 

50 50 41 50 

51.2 50.5 46.2 39.8 

22 (44%) 

4696 

21 (42%) 

44% 

16 (34%) 

35 % 

9 (18%) 

22% 

a Excludesanimalsdying prior to oneyear 

theliverindosedmalegroups,andadecreased intake) and the associated body weight gain depresincidenceofeosinophilicfociin 25 mg/kgfemale sion. Additionally, dietary restriction and associated 

mice.Therewerealsodecreasedincidencesof body weight depression inhibits the development of 

common spontaneously occurring lesions in male anda variety of neoplasms, including leukemia (Albanes, 

femalemicereceivingscopolaminehydrobromide 1987; Weindruch and Walford, 1988; Boissonneult, 

trihydrate.Theseincludedpancreaticislethyper- 1991). 

plasia (males), alveolar epithelial hyperplasia (males), 

kidneynephropathy,pituitaryglandparsdistalis 

hyperplasia(females),bonemarrowmyelofibrosis 

(females),uterinecystichyperplasia,andhematopoieticcellproliferationinthespleen(females). 

Decreasedincidencesofthesespontaneouslesions 

were most likely a result of lower body weights in 

dosed groups. 

The results of the self-selection studies ofRoss et al. 

(1983a,b) support the view that reduced body weight 

gains in rodents are associated with lower incidences 

of spontaneous disease. The body weights of mature 

rats correlated linearly with the incidence of spontaneousneoplasms.Thoseratsthatgrewrapidly 

developed anterior pituitary neoplasms more readily 

than slower growing rats. Tucker (1979) observed a 

Chronicrepeatedadministrationofscopolamine 

hydrobromide trihydrate marginally affected performancebyratsin three ofthefiveneurobehavioral 

testsemployedduringthisstudy.Theseresultsare 

the only observationsdocumentingtheeffects of 

chronic scopolamine hydrobromide trihydrate exposure on rodent behavior. 

Althoughhyperactivityoccurredin 25 mg/kgrats 

fromday 1 (females)andonday 180 (males), only 

the increased horizontal activity in females on treatment days 90, 180, and 360 was statistically significant.Scopolamine-inducedhyperactivity(ambulation) after acute exposures in rats has been demonstratedbyseveralinvestigatorsusinglowerdoses 

(0.25 to 4.0 mg/kg) and different routes of adminisdecreased incidence of pituitary gland neoplasms withtration (subcutaneous and intraperitoneal) (Reiter and 

dietaryrestriction (20% lessfeedthan ad libitum McPhail, 1982; Sanberg et al., 1987; Crofton et al., 
60 

1991). Other investigators have demonstratedboth 

scopolamine-induced increases (Bauer, 1984) and 

decreases (Horsburgh and Hughes, 1981) in vertical 

activity (rearings) following intraperitoneal exposure. 

There was a nonsignificant tendency for female rats 

receiving 5 and 25 mg/kg scopolamine hydrobromide 

trihydrate on days 90 through 360 to showan 

increased rearing response during this study. Reiter 

and McPhail (1982) describe a variety of operational 

factors which influence motor activity including the 

decreased tendency for scopolamine to induce hyperactivity as the environmental complexity (novel 

stimuli) is increased. This concept was supported by 

Renner et al. (1992), whonoteddecreased preferences for novelty by scopolamine-treated rats in an 

open field arena. MacMahon et al. (1981) concluded 

that scopolamine has adverse affects in rats by 

pairing treatments (unconditionedstimulus)with 

novel stimuli (conditioned stimulus). Their conclusion was based on observation that the conditioned 

stimulus was avoided during a free choice test. 

The startle response to tactile stimulation was attenuated in female rats receiving 5 and 25 mg/kg scopolamine hydrobromide trihydrate. This effect occurred 

as an isolated incidence (90 daytestinterval)and 

could not be attributed to any effects on musculature 

(grip strength test). A depressed startle response 

couldpossibly ascribed be to reduced 

attentionhigilance, an effect sometimes associated 

with scopolamine exposures (Wesnes andRevell, 

1984; Wesnesand Warburton, 1984; Lydonand 

Nakajima, 1992). 

Although rats receiving 25 mg/kg scopolamine 

hydrobromide trihydrate on study days 180 through 

720 tended to score lower passive avoidance latency 

scores, this effect was significantonly on study 

day 180 in 25 mg/kg males. In rats, lower passive 

avoidance latency scores following acute scopolamine 

hydrobromide trihydrate treatment at lower doses 

(0.1 to 1 .O mg/kg) have beenpreviously reported 

(Blozovskiand Hennocq, 1982; Elrod an 

Buccafusco, 1988). These findings may be indicators 

of learning and memory deficits (Cabeand 

Scopolamine Hydrobromide Trihydrate, NTP TR 445 

Eckerman, 1982). Scopolamine-treated humans

(Drachman, 1978; Caine et al., 1981) and other 

mammalsincluding rodents (Lenkgre et al., 1988) 

are models for amnesia in the development of nootropic drugs. Nootropic drugs enhance memory and 

attenuate experimental amnesia (Ledgre et al., 

1988). There are reports that experimentally induced 

scopolamine deficits in learning and memory become 

more pronounced with age in humans (Flicker et al., 

1992; Molchan e? al., 1992). While our results at 

studyday 720 suggest depressed latency scores at 

lower doses, they are not statistically significant. 

Theseneurobehavioral findings do not support a 

robust scopolamine effect on passive avoidance 

responses. A possible explanation may involve an 

accommodation or adjustment of the nervous system 

to chronic repeated exposures. Tilson and Harry 

(1982) note that the Fischer rat strain tends to more 

efficiently acquire or learn avoidance behavior and is 

resistant to scopolamine-induced decrements in this 

behavior. It should also be noted that scopolamine 

affectsboththe peripheral and central nervous 

systemsandthat higher doses of the drug might 

induce conflicting results on the peripheral nervous 

system(Andrews e? al., 1994). In this regard this 

study is unique in that it uses chronic oral gavage 

exposure and comparatively high doses. The lack of 

significant clinical observations and the low scopolamine serum concentrations do not support the view 

that scopolamine-induced peripheral nervous system 

effects couldhave altered avoidance behavior. 

Finally, the absence of a scopolamine effect on 

pawlicklatency suggests that scopolamine-induced 

analgesia could not have disrupted avoidance behavior by decreasing latency scores. 

CONCLUSIONS 

Under the conditions of these 2-year gavage studies, 

there was no evidence of carcinogenicactivity* of 

scopolamine hydrobromide trihydrate in male or 

female F344/N rats or B6C3F1 mice administered 1, 

5, or 25 mg/kg. 

* Explanation of Levels of Evidence of Carcinogenic Activity is on page 10. A summary of the Technical Reports Review Subcommittee 

comments and the public discussion on this Technical Report appears on page 12. 
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67 

APPENDIX A 

SUMMARY OF LESIONS IN MALE RATS 

IN THE 2-YEAR GAVAGE STUDY 

OF SCOPOLAMINE HYDROBROMIDE TRIHYDRATE 

TABLEA1 Summary of the Incidence of Neoplasms in Male Rats 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........... 69 

TABLEA2 Individual Animal Tumor Pathology of Male Rats 

TABLEA3 Statistical Analysis of Primary Neoplasms in Male Rats 

TABLEA4a Historical Incidence of Pituitary Gland (pars Distalis) Adenoma 

TABLEA4b Historical Incidence of Mononuclear Cell Leukemia 

TABLEA5 Summary of the Incidence of Nonneoplastic Lesions in Male Rats 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........... 74 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........... 96 

in Male F344/N Rats Administered Water by Gavage ...................... 101 

in Male F344/N Rats Administered Water by Gavage ...................... 101 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........... 102 
68 Scopolamine Hydrobromide Trihydrate, NTP TR 445 
Lesions in Male Rats 69 

TABLEA1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydratea 

VehicleControl 1 mgkg 5 mgkg 25 mgkg 

Disposition Summary 

Animals initially in study 

IS-Month interim evaluation 

Early deaths 

Accidental deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

Endocrine System 

Adrenal medulla 

Pheochromocytoma benign 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

Genital System 

Preputial gland 

Adenoma 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

IntegumentarySystem 

Skin 

Squamous cell papilloma 

Musculoskeletal System 

Bone 

Osteosarcoma 

Systemic Lesions 

Multiple organsb 

Leukemia mononuclear 

60 

10 

2 

23 

5 

20 

60 

(10) 

(10) 

(10) 

(10) 

5 (50%) 

4 (40%) 

60 

10 

2 

30 

4 

14 

60 

(10) 

(10) 

6 (60%)

2 (20%) 

60 

10 

1 

20 

7 

22 

60 

(9) 

1 (11%) 

(10) 

6 (60%)

3 (30%) 

60 

10 

1 

17 

4 

28 

60 

(10) 

(10) 

6 (60%)

3 (30%) 

(9) 

1 (11%) 
70 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA1 

Summary of the Incidence of Neoplasms in Male Rats in the2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

15-Month Interim Evaluation (continued) 

Systems Examined With No Neoplasms Observed 

Alimentary System 

Cardiovascular System 

General Body System 

HematopoieticSystem 

Nervous System 

Respiratory System 

Special Senses System 

Urinary System 

2-Year Study 

Alimentary System 

Esophagus 

Intestine large, colon 

Leiomyoma 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Osteosarcoma, metastatic, uncertain primary 

site 

Mesentery (13)

Carcinoma, metastatic, islets, pancreatic 1 (8%) 

Osteosarcoma, metastatic, uncertain primary 

site 1 (5%) 

Pancreas (50) 

Osteosarcoma, metastatic, uncertain primary 

site 1 (2%) 

Acinus, adenoma 

Salivary glands (50)

Schwannoma malignant 

Tongue 

Squamous cell papilloma 

Tooth (1)

Gingiva, squamous cell carcinoma 1(100%) 

Cardiovascular System 

Heart 

Schwannoma NOS 

Endocrine System 

Adenoma 

Adrenal cortex (50) 

1 (2%) 

(50) (50) 

Pheochromocytoma benign 

Adrenal medulla (50) 

8 (16%) 

(50) 

11 (22%) 

(50) 

9 (18%) 

Bilateral, pheochromocytoma benign 3(6%) 2 (4%) 3(6%) 

2 (4%) 

(49) 

(1) 

1(100%) 

(50) 

(50) 

8 (16%) 

1 (2%) 
Lesions in Male Rats 71 

TABLEA1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 25 mgkg 

2-Yeur Study (continued) 

Endocrine System (continued) 

Islets, pancreatic 

Adenoma 

Carcinoma 

Parathyroid gland 

Adenoma 

Pituitary gland 

Pars distalis, adenoma 

Pars intemedia, adenoma 

Thyroid gland 

Bilateral, Ctell, adenoma 

C-cell, adenoma 

Ctell, carcinoma 

Follicular cell, adenoma 

Follicular cell. carcinoma 

General Body System 

None 

GenitalSystem 

Epididymis 

Osteosarcoma, metastatic, uncertain primary 

site 

Preputial gland 

Adenoma 

Carcinoma 

Testes 

Osteosarcoma, metastatic, uncertain primary 

site 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

HematopoieticSystem 

Lymph node 

Mediastinal, osteosarcoma, metastatic, 

uncertain primary site 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Fibroma 

Hemangiosarcoma 

Osteosarcoma, metastatic, uncertain primary 

site 

Thymus 

Osteosarcoma, metastatic, uncertain primary 

site 

Thymoma malignant 

(50) 

3(6%) 

1 (2%) 4(8%) 

1 (2%) 

3 (6%) 2 (4%) 

1 (2%) 

(50) 

(50) 

3 (6%) 

(50) 

38 (76%) 

5 (10%) 

(5) 

(50) 

(50) 

(50) 

(46) 

1 (2%) 

1 (2%) 
72 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA1 

Summary of the Incidence of Neoplasms in Male Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

2-Year Study (continued) 

IntegumentarySystem 

Mammary gland (48) (49) (49)

Fibroadenoma 8 (17%) 3 (6%) 2 (4%) 

Fibroadenoma. multiple 1 (2%)

Skin (50) Basal cell adenoma 1 (2%) 

Fibroma 3 (6%) 2 (4%)

Fibroma,multiple 1 (2%) 

Fibrous histiocytoma 

Keratoacanthoma 2 (4%) 

Keratoacanthoma,multiple 

Sarcoma 

Squamous cell carcinoma 

Squamous cell papilloma 

Sebaceous gland, adenoma 

Musculoskeletal System 

Bone (50) (50) (50) (50)

Osteosarcoma 

Osteosarcoma, metastatic, uncertain primary 

Carcinoma, metastatic, islets, pancreatic 

Skeletal muscle 

Sarcoma 

site 

1 (2%) 

(1) 

1 (100%) 

1 (100%) 

1 (2%) 

(1) (1) 

1 (2%) 

1 (100%) 

Spinal cord 

Brain 

Nervous System 

Astrocytoma NOS 

Lung 

Respiratory System 

Squamous cell carcinoma. metastatic, tooth 

Osteosarcoma, metastatic, bone 

Alveolarlbronchiolar carcinoma 

Alveolarlbronchiolar adenoma 

(50) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

Eye 

Special Senses System 

Carcinoma 

Zymbal's gland 

Adenoma 

Harderian gland 
Lesions 73 in Male Rats 

TABLEA1 

Summary of the Incidence of Neoplasms in MaleRats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

&Year Study (continued) 

Urinary System 

Kidney 

Fibroma 

Osteosarcoma, metastatic, uncertain primary 

site 

Renal tubule, adenoma 

Renal tubule, carcinoma 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma NOS 

Neoplasm Summary 

Total animals with primary neoplasms' 

15-Month interim evaluation 

2-Year study 

Total primary neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with benign neoplasms 

15-Month interim evaluation 

2-Year study 

Total benign neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals with malignant neoplasms 

15-Month interim evaluation 

2-Year study 

Total malignant neoplasms 

15-Month interim evaluation 

2-Year study 

Total animals wi* metastatic neoplasms 

2-Year study 

Total metastatic neoplasms 

2-Year study 

Total animals with malignant neoplasms 

of uncertain primary site 

2-Year study 

Total animals with uncertain neoplasmsbenign or malignant 

2-Year study 

Total uncertain neoplasms 

2-Year study 

1 (2%) 

1 (2%) 

(50) 

(50) 

33(66%) 

1(2%) 

9 

49 

10 

145 

9 

48 

9 

101 

1 

38 

1 

43 

1 

1 

1 

3 

(50) 

(50) 

21(42%) 

1 (2%) 

8 

49 

8 

132 

8 

49 

8 

104 

27 

27 

2 

3 

1 

4 

(50) 

26(52%) 

4(8%) 

10 

49 

11 

132 

10 

48 

11 

91 

30 

35 

1 

10 

1 

6 

17 

(50) 

24(48%) 

3 (6%) 

9 

48 

14 

112 

9 

48 

13 

80 

1 

28 

1 

29 

1 

1 

3 

8 

a 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 
74 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl 

Number of Dayson Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Mesentery 

Pancreas 

Acinus,adenoma 

Salivary glands 

Schwannoma malignant 

Stomach, forestomach 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adenoma 

Adrenal medulla 

Pheochromocytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

Ccell, carcinoma 

Follicular cell, adenoma 

Follicular cell, carcinoma 

General Body System 

None 

+: Tissue examined microscopically 

A:Autolysis precludesexamination 

2444455556666666666666667 

7347902680344455566778890-

3031283610925956603136682 

0000000000000000000000000 

3433252201241040430551103 

3981489067870228309478949 

......................... 

......................... 

++++++++++++++++++++++M++ 

......................... 

......................... 

......................... 

......................... 

......................... 

+ + + ......................... 

......................... X 

......................... 

......................... 

......................... 

......................... 

......................... X 

......................... X X 

X 

......................... 

++++++++M++++++M++++M+++M 

......................... x x x x X X xxx ......................... X 

X X 

M:Missingtissue X: Lesionpresent 

I:Insufficienttissue Blank:Not examined 
Lesions in Male Rats 75 

TABLEA2 

Individual AnimalTumor Pathology of Male Rats in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) ' 

7777777777777777777777777 

Number of Days on Study 0012222222222222222222222 

2630066666677777777777777 

0000000000000000000000000 Total 

Carcass ID Number 5612312445500111223344445 Tissues/ 

0057263583517234254701469 Tumors 

AlimentarySystem 

Esophagus ......................... 50 

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 

Hepatocellular carcinoma X 1 

Hepatocellular adenoma X 1 

Mesentery + + + + 7 

Pancreas ......................... 50 

Acinus, adenoma X 1 

Salivary glands ......................... 50 

Schwannoma malignant 1 

Stomach, forestomach ......................... 50 

Stomach, glandular ......................... 50 

Cardiovascular System 

Blood vessel ......................... 50 

Heart ......................... 50 

Endocrine System 

Adrenal cortex ......................... 50 

Adenoma 1 

Adrenal medulla ......................... 50 

Pheochromocytoma benign X x x X xx 8 

Bilateral, pheochromocytoma benign X X 3 

Islets, pancreatic ......................... 50 

Adenoma X X X 3 

Parathyroid gland ......................... 46 

Pituitary gland +++++++++++++++++++M+++++ 49 

Pars distalis, adenoma x xx x xx X x xx 19 

Thyroid gland ......................... 50 

Ctell. adenoma 1 

C-cell, carcinoma X 1 

Follicular cell, adenoma X 3 

Follicular cell, carcinoma X 1 

General Body System 

None 
76 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Y-r Gavage Study 

of Scopolamine HydrobromideTrihydrate:VehicleControl (continued) 

Number of Days on Study 

Carcass ID Number 

Genital System 

Epididymis 

Preputial gland 

Adenoma 

Prostate 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

HematopoieticSystem 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

IntegumentarySystem 

Mammary gland 

Fibroadenoma 

Skin 

Fibroma 

Fibroma, multiple 

Keratoacanthoma 

Keratoacanthoma, multiple 

Sarcoma 

Squamous cell carcinoma 

Sebaceous gland, adenoma 

Musculoskeletal System 

Bone 

Osteosarcoma 

Skeletal muscle 

Sarcoma 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Alveolar/bronchiolar carcinoma 

Osteosarcoma, metastatic, bone 

Nose 

Trachea 

~~~ ~ 

2444455556666666666666667 

7347902680344455566178890 

3031283610925956603136682 

00000000~00000000000000000 

3433252201241040430551103 

3981489067870228309478949 

......................... 

......................... X 

......................... 

......................... 

......................... x x x x x x x x x x x x x x x x x 

X 

......................... + + + + ......................... 

......................... 

......................... 

++++++++++++++++++++M+++M 

+++++++++++M+++++++++++++ 

X 

......................... x x X 

X 

X 

......................... X 

+ 

X 

......................... + + + + ++ 

+ + + + ++ 

......................... X 

X 

......................... 

......................... 
Lesions in Male Rats 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

Number of Days on Study 

Carcass ID Number 

Genital System 

Epididymis 

Preputial gland 

Adenoma 

Prostate 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

HematopoieticSystem 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

IntegumentarySystem 

Mammary gland 

Fibroadenoma 

Skin 

Fibroma 

Fibroma, multiple 

Keratoacanthoma 

Keratoacanthoma, multiple 

Sarcoma 

Squamous cell carcinoma 

Sebaceous gland, adenoma 

Musculoskeletal System 

Bone 

Osteosarcoma 

Skeletal muscle 

Sarcoma 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Alveolarlbronchiolar carcinoma 

Osteosarcoma, metastatic. bone 

Nose 

Trachea 

7777777777777777777777777 

001222222222.2222222222222 

2630066666677777777777777 

0000000000000000000000000 Total 

5612312445500111223344445 Tissues/ 

0057263583517234254701469 Tumors 

......................... 50 

......................... 50 

X X 3 

......................... 50 

......................... 50 

......................... 50 

x xx x xx x x x x x x x x xxxxxx 38 

X X X X 5 

......................... 50 

+ 5 

......................... 50 

......................... 50 

......................... 50 

+++++++++++++++++++M++++M 46 

++++++++++++++M++++++++++ 48 

X X 3 

......................... 50 

X 4 

1 

X X X 3 

X 1 

1 

X 1 

X 1 

......................... 50 

1 

1 

1 

......................... 50 

+ ++ 9 

+ ++ 9 

......................... 50 

1 

1 

......................... 50 

......................... 50 
78 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

2444455556666666666666667 

Number of Days on Study 7347902680344455566778890 

3031283610925956603136682 

0000000000000000000000000 

Carcass ID Number 3433252201241040430551103 

3981489067870228309478949 

Special Senses System 

Ear + 

EYe Harderian gland + 

Adenoma X 

Urinary System 

~ ~~ 

Kidney ......................... Renal tubule, adenoma 

Renal tubule, carcinoma 

Urinary bladder ......................... 

SystemicLesions 

~~ 

Multiple organs ......................... Leukemia mononuclear x x x x x xx xx x xx x x x 

Mesothelioma NOS X 
Lesions in Male Rats 79 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

Number of Days on Study 

Carcass ID Number 

~~ ~~ ~ 

Special Senses System 

Ear 

EYe Harderian gland 

Adenoma 

Urinary system 

~~~ 

Kidney 

Renal tubule, adenoma 

Renal tubule, carcinoma 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma NOS 

7777771771777771777177777 

0012222222222222222222222 

2630066666677771117771777 

0000000000000000000000000 Total 

5612312445500111223344445 Tissues/ 

0057263583517234254101469 Tumors ~ 

+ + 

+ 

......................... 50 

X 1 

X 1 

......................... 50 

......................... 50 

X x x x x x x x x x x x x x x x 33 

1 
80 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathology of Male Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 1 mg/kg 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Mesentery 

Carcinoma, metastatic, islets, 

pancreatic 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tongue 

Squamous cell papilloma 

Cardiovascular System 

Blood vessel 

Heart 

EndocrineSystem 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Carcinoma 

Parathyroid gland 

Adenoma 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Epididymis 

Preputial gland 

Adenoma 

Carcinoma 

Prostate 

2444555555555556666666666 

8049003346688891335566667 

1462443300135922126800691 

0010000100000000101111010 

6916689169797879071000818 

3197568765865074794935558 

......................... 

......................... 

.......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

X 

+ ++ 

......................... 

.......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... X xx X 

......................... 

......................... 

+++++++++++++++++++++M+++ 

X X xx xxxx ......................... X X 

......................... 

......................... 

X 

......................... 
Lesions in Male Rats 81 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: 1mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Mesentery 

Carcinoma, metastatic, islets, 

pancreatic 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tongue 

Squamous cell papilloma 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Carcinoma 

Parathyroid gland 

Adenoma 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Epididymis 

Preputial gland 

Adenoma 

Carcinoma 

Prostate 

6666611111111111111111111 

1178800001122222222222222 

8888822263511111111111111 

0011100011000000000111111 Total 

6900189921166111889000111 Tissues/ 

2062122308019236311014023 Tumors 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 .,

A 1 

A 2 

+ + + ++ + ++ + + 13 

X 1 

......................... 50 

++++++++M++++++++++++++++ 49 

......................... 50 

.......................... 50 

+ 1 

X 1 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

xx x xx x x 11 

X X 2 

......................... 50 

X X 2 

X 1 

+++++++++++++++M+M+M++++M 45 

X 1 

......................... 49 

x xxx x xx xx 11 

......................... 50 

X X 4 

X X 2 

......................... 50 

......................... 50 

X 1 

1 

......................... 50 
82 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Gavage Study 

of Scopolamine HydrobromideTrihydrate: 

Number of Days on Study 

Carcass ID Number 

Genital System (continued) 

Seminal vesicle , 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Fibroma 

Thymus 

Integumentary System 

Mammary gland 

Fibroadenoma 

Skin 

Fibroma 

Keratoacanthoma 

Squamous cell papilloma 

Musculoskeletal System 

Bone 

Osteosarcoma 

Skeletal muscle 

Carcinoma, metastatic, islets, 

pancreatic 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Osteosarcoma, metastatic, bone 

Nose 

Trachea 

1 mg/kg (continued) 

2444555555555556666666666 

8049003346688891335566667 

1462443300135922126800691 

0010000100000000101111010 

6916689169797879071000818 

3197568765865074’794935558 

......................... 

......................... xxx xx xxxxxxx x x

X X X X X 

......................... + + + ......................... 

......................... 

......................... 

+++M++++++++++++++++M++++ 

++M++++M+++++++++++++++++ 

X xx X ......................... X X 

X 

......................... X 

......................... + 

+ 

......................... X 

......................... 

......................... 
Lesions in MaleRats 83 

TABLEA2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 1 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Genital System (continued) 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

HematopoieticSystem 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Fibroma 

Thymus 

IntegumentarySystem 

MammaIy gland 

Fibroadenoma 

Skin 

Fibroma 

Keratoacanthoma 

Squamous cell papilloma 

Musculoskeletal System 

Bone 

Osteosarcoma 

Skeletal muscle 

Carcinoma, metastatic, islets, 

pancreatic 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Osteosarcoma, metastatic, bone 

Nose 

Trachea 

6666677777777777777777777 

7778800001122222222222222 

8888822263577777777777777 

0011100011000000000111111 Total 

6900189921766777889000111 Tissues/ 

2062122308019236377014023 Tumors 

......................... 50 

......................... 50 

X xxxx xxxx xxxx xx X 30 

X X X X X xx 12 

......................... 50 

++ ++ + 8 

......................... 49 

......................... 50 

......................... 50 

X 1 

++M+++++M++++++++++++++++ 46 

......................... 48 

X X X X 8 

......................... 50 

X 3 

X X X 3 

X X 3 

......................... 50 

1 

+ 1 

X 1 

......................... 50 

++ + ++ + 7 

++ + ++ + 7 

......................... 50 

1 

......................... 50 

......................... 50 
84 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromide Trihydrate: 1mg/kg (continued) 

2444555555555556666666666 

Number of Days on Study 8049003346688891335566667 

1462443300135922126800691 

0010000100000000101111010 

Carcass ID Number 6916689169797879071000818 

3197568765865074794935558 

Special Senses System 

Zymbal’s gland 

EYe 

Carcinoma 

+ 

X 

+ 

X ~~~ ~~~ 

Urinary System 

Kidney ......................... Fibroma 

Urinary bladder ......................... 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear xx ......................... x x x x 

Mesothelioma NOS X 
Lesions in Male Rats 85 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: 1mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Special Senses System 

Eye

Zymbal’s gland 

Carcinoma 

Urinary System 

Kidney 

Fibroma 

Urinary bladder 

SystemicLesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma NOS 

6666611111111111111111111 

1118800001122222222222222 

8888822263511111111111111 

0011100011000000000111111 Total 

6900189921166111889000111 Tissues/ 

2062122308019236311014023 Tumors 

+ 1 

2 

2 

......................... 50 

X 1 

......................... 50 

......................... 50 

xxxxx x xx xx x x x 21 

1 
86 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular adenoma 

Osteosarcoma, metastatic, uncertain 

primary site 

Mesentery 

Osteosarcoma, metastatic, uncertain 

primary site 

Pancreas 

Osteosarcoma, metastatic, uncertain 

primary site 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tooth 

Gingiva, squamous cell carcinoma 

Cardiovascular System 

Blood vessel 

Heart 

Schwannoma NOS 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Bilateral, C-cell, adenoma 

C-cell, adenoma 

C-cell, carcinoma 

Follicular cell, adenoma 

General Body System 

None 

1455555666666666666666666 

I667789022333444456678888 

8356696957111366960631666 

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 

4275532344377844322722366 

9604777328517004495384105 

......................... 

......................... 

......................... 

......................... 

......................... 

+++++++++++++++++M+++++++ 

......................... 

......................... X 

+++ +++ ++ + ++ + 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... X xxxx 

X X 

......................... 

......................... 

......................... X x xx x x ......................... X 

X 
Lesions in Male Rats 87 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular adenoma 

Osteosarcoma, metastatic, uncertain 

primary site 

Mesentery 

Osteosarcoma,.metastatic, uncertain 

primary site 

Pancreas 

Osteosarcoma, metastatic, uncertain 

primary site 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tooth 

Gingiva, squamous cell carcinoma 

Cardiovascular System 

Blood vessel 

Heart 

Schwannoma NOS 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Bilateral, C-cell, adenoma 

C-cell, adenoma 

C-cell, carcinoma 

Follicular cell, adenoma 

General Body System 

None 

7777777717771777777177171 

0112222222222222222222222 

6336666666677777777777777 

1111111111111111111111111 Total 

3552456777123344556666671 Tissues/ 

8282169468936936011261825 Tumors 

......................... 50 

......................... 50 

......................... 50 

M++++++++++++++++++++++++ 49 

......................... 50 

......................... 49 

......................... 50 

......................... 50 

X 2 

X 1 

+ ++ + ++++ ++ 22 

X 1 

......................... 50 

X 1 

......................... 50 

......................... 50 

......................... 50 

+ 1 

X 1 

......................... 50 

......................... 50 

X 1 

......................... 50 

......................... 50 

X X X X 9 

X 3 

......................... 50 

X 1 

++++++M++++M+++MM++++++++ 46 

......................... 50 

x xx xx x X 13 

......................... 50 

1 

X 2 

X X X 3 

X 1 
88 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

~ _____ ~~~___~ 

Genital System 

Epididymis 

Osteosarcoma, metastatic, uncertain 

primary site 

Penis 

Preputial gland 

Adenoma 

Prostate 

Seminal vesicle 

Testes 

Osteosarcoma, metastatic, uncertain 

primary site 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

HematopoieticSystem 

Bone marrow 

Lymph node 

Mediastinal, osteosarcoma, 

metastatic, uncertain primary site 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Osteosarcoma, metastatic, uncertain 

primary site 

Thymus 

Osteosarcoma, metastatic, uncertain 

primary site 

IntegumentarySystem 

Mammary gland 

Fibroadenoma 

Skin 

Fibroma 

Fibrous histiocytoma 

Keratoacanthoma 

Musculoskeletal System 

Bone 

Osteosarcoma 

Skeletal muscle 

Osteosarcoma, metastatic, uncertain 

primary site _.

NervousSystem 

Brain 

Astrocytoma NOS 

Peripheral nerve 

Spinal cord 

1455555666666666666666666 

7667789022333444456678888 

8356696957111366960631666 

1111111111111111111111111 

4275532344377844322722366 

9604777328517004495384105 

......................... 

+ ......................... X X X 

......................... 

......................... 

......................... 

xx x xxxx X xxxxx X 

x x x x x X 

......................... + + + 

......................... 

......................... 

......................... 

++++M++++M+++++++++++++++ 

......................... X 

......................... X 

X 

X 

......................... X 

......................... 

++ ++ 

++ ++ 
Lesions in Male Rats 89 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Genital System 

Epididymis 

Osteosarcoma. metastatic, uncertain 

primary site 

Penis 

Preputial gland 

Adenoma 

Prostate 

Seminal vesicle 

Testes 

Osteosarcoma. metastatic, uncertain 

primary site 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

Hematopoietic System 

Bone marrow 

Lymph node 

Mediastinal, osteosarcoma, 

metastatic, uncertain primary site 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Osteosarcoma, metastatic, uncertain 

primary site 

Thymus 

Osteosarcoma. metastatic, uncertain 

primarysite 

Integumentary System 

Mammary gland 

Fibroadenoma 

Skin ' 

Fibroma 

Fibrous histiocytoma 

Keratoacanthoma 

Musculoskeletal System 

Bone 

Osteosarcoma 

Skeletal muscle 

Osteosarcoma, metastatic, uncertain 

primary site 

Nervous System 

Brain 

Astrocytoma NOS 

Peripheral nerve 

Spinal cord 

7777777777777777777777777 

,0112222222222222222222222 

6336666666677777777777777 

1111111111111111111111111 Total 

3552456777723344556666677 Tissues1 

8282169468936936011267825 Tumors 

......................... 50 

X 1 

1 

+++++++++++++++++++M+++++ 49 

3 

......................... 50 

......................... 50 

......................... 50 

X 1 

x x x x x x x x x x xx x xxxxxxxx 35 

x x X X 11 

......................... 50 

+ + 5 

X 1 

......................... 50 

......................... 50 

......................... 50 

X 1 

......................... 48 

X 1 

++++++++++M++++++++++++++ 49 

X X 3 

......................... 50 

X X 3 

1 

x x 3 

......................... 50 

1 

+ 1 

X 1 

......................... 50 

X 1 

+ + , 6

+ + 6 
90 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Respiratory System 

Lung 

Alveolarhronchiolar adenoma 

Nose 

Trachea 

Special Senses System 

Ear 

EY e 

Harderian gland 

Zymbal's gland 

Carcinoma 

Urinary System 

Kidney 

Osteosarcoma, metastatic, uncertain 

primary site 

Renal tubule, carcinoma 

Urinary bladder 

~ ~~~ ~ 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma NOS 

1455555666666666666666666 

7661789022333444456678888 

8356696957111366960631666 

1111111111111111111111111 

42'75532'344377844322722366 

9604777328517004495384105 

......................... 

......................... 

++++++++++++++++++++M++++ 

+ 

+ 

+ 

X 

......................... 

X 

......................... 

......................... x x xx xxxxxxxxxx 

X xx 
Lesions in Male Rats 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

RespiratorySystem 

Lung

Alveolarlbronchiolar adenoma 

Nose 

Trachea 

~~~ ~ 

Special Senses System 

Ear 

EYe 

Harderian gland 

Zymbal’s gland 

Carcinoma 

Urinary System 

Kidney 

Osteosarcoma, metastatic, uncertain 

primary site 

Renal tubule, carcinoma 

Urinary bladder 

SystemicLesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma NOS 

1111111111111111111111171 

0112222222222222222222222 

6336666666611177711771117 

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Total 

3552456111123344556666611 Tissues1 

8282169468936936011261825 Tumors 

......................... 50 

X 1 

......................... 50 

......................... 49 

t 

+ 

X 

......................... 50 

X 1 

1 

......................... 50 

......................... 50 

xx x X xxx x xxxx 26 

X 4 
92 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromide Trihydrate: 25 mg/kg 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Intestine large, colon 

Leiomyoma 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular adenoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tooth 

Gingiva, squamous cell carcinoma 

Cardiovascular System 

Blood vessel 

Heart 

EndocrineSystem 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Pars intermedia, adenoma 

Thyroid gland 

C-cell, adenoma 

General Body System 

None 

GenitalSystem 

Epididymis 

Preputial gland 

Adenoma 

Prostate 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

113455555555666666666777.7 

2450014678885677788891222 

6414460554996633945623666 

2211222222122212212112112 

0099011322833192382892890 

6368430695383692111648450 

......................... 

......................... X 

......................... 

......................... 

......................... 

++++++++++++++++++++M++++ 

+++++++++++++++A+++++++++ 

......................... X 

++ + + + + + ++ ......................... 

M++++++++++++++++++++++++ 

........................... 

......................... 

......................... 

......................... 

......................... 

......................... X X X 

......................... 

......................... 

......................... xx x x X x x 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... xxx x xxxxxxxxxxxxx 

X X 
Lesions in Male Rats 93 

TABLEA2 

Individual AnimalTumor Pathology of Male Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mglkg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Intestine large. colon 

Leiomyoma 

Intestine large, rectum 

Intestine large. cecum 

Intestine small, duodenum 

Intestine small. jejunum 

Intestine small, ileum 

Liver 

Hepatocellular adenoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tooth 

Gingiva, squamous cell carcinoma 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Bilateral, pheochromocytoma benign 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Pars intermedia, adenoma 

Thyroid gland 

C-cell, adenoma 

General Body System 

None 

Genital System 

Epididymis 

Preputial gland 

Adenoma 

Prostate 

Seminal vesicle 

Testes 

Bilateral, interstitial cell, adenoma 

Interstitial cell, adenoma 

1111777111111111111171111 

2222222222222222222222222 

6666666666611111111171117 

2222222222211111122222222 Total 

0111122233388899900011234 Tissues/ 

8518934625958912115914140 Tumors 

......................... 50 

.......................... 50 

1 

......................... 50 

......................... 50 

......................... 50 

......................... 49 

......................... 49 

......................... 50 

X 2 

+ + +++ 14 

......................... 50 

......................... 49 

......................... 50 

......................... 50 

+ 1 

X 1 

++++++++++++++++++++++++M 49 

......................... 50 

......................... 50 

......................... 50 

xx X X X 8 

X 1 

......................... 50 

+++++++++M+++++++++++++++ 49 

......................... 50 

xx X 10 

X 1 

......................... 50 

X 1 

......................... 50 

......................... 50 

X X 2 

......................... 50 

......................... 50 

......................... 50 

x x x x x x x x x x x x x x xx x x x x x x x x 41 

X 3 
94 Scopolamine Hydrobromide Trihydrate, NTP TFt 445 

TABLEA2 

Individual Animal Tumor Pathology of Male Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

HematopoieticSystem 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma 

Thymus 

Thymoma malignant 

IntegumentarySystem 

Mammary gland 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Basal cell adenoma 

Fibroma 

Fibroma, multiple 

Keratoacanthoma 

Squamous cell papilloma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Squamous cell carcinoma, metastatic, 

tooth 

Nose 

Trachea 

Special Senses System 

Ear 

EYe 

Zymbal’s gland 

Carcinoma 

Urinary System 

Kidney 

Urinary bladder 

SystemicLesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma NOS 

1134555555556666666667177 

2450014678885617788891222 

6414460554996633945623666 

2211222222122212212112112 

0099011322833192382892890 

6368430695383692111648450 

......................... + ++ +++ + + + ......................... 

......................... 

......................... 

+++++++M++++M++++++++M+++ A 

++++++++++++++++++++M++++ 

......................... X 

X 

X 

......................... 

......................... + + + + 

+ + + + 

......................... 

......................... 

......................... 

+ + 

X X 

......................... 

......................... 

......................... X xxxx x xxxxxx 
Lesions in Male Rats 95 

TABLEA2 

Individual Animal Tumor Pathologyof Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma 

Thymus 

Thymoma malignant 

Integumentary System 

Mammary gland 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Basal cell adenoma 

Fibroma 

Fibroma, multiple 

Keratoacanthoma 

Squamous cell papilloma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Squamous cell carcinoma, metastatic, 

tooth 

Nose 

Trachea 

Special Senses System 

Ear 

EY e Zymbal’s gland 

Carcinoma 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

Mesothelioma NOS 

1711117171111171171111171 

2222222222222222222222222 

6666666666611111111711111 

2222222222211111122222222 Total 

0111122233388899900011234 Tissues/ 

8578934625958912715914140 Tumors 

......................... 50 

9 

......................... 49 

......................... 50 

......................... 50 

X 1 

......................... 41 

1 

......................... 49 

X X 2 

X 1 

......................... 50 

1 

X X 2 

1 

X 2 

X 1 

......................... 50 

......................... 50 

+ 5 

+ 5 

......................... 50 

X 1 

......................... 50 

......................... 50 

+ + 2 

+ 1 

2 

2 

......................... 50 

......................... 50 

......................... 50 

x xx x xxxxx X 24 

X X X 3 
96 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA3 

Statistical Analysis of Primary Neoplasms in MaleRats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

VehicleControl 1 mgkg 5 mgkg 25 mgkg 

AdrenalMedulla:BenignPheochromocytoma 

Overall ratea 11/50 (22%) 13/50 (26%) ., 12/50 (24%) 9/50 (18%) 

Adjusted rateb 41.4%" 57.0% 37.7% 29.0% 

Terminal rate' 5/20 (25 %) 6/14 (43%) 4/22 (18%) 7/28 (25%) 

First incidence (days) 673 585 589 656 

Life table testd P=0.057N P=O.156 P=O.532 P=0.189N 

Logistic regression testd P=0.162N P10.243 P=O.526 P=0.315N 

Cochran-Armita e testd P=0.248N

Fisher exact test % P-0.408 P=O.5OO P=0.402N 

Liver: HepatocellularAdenomaorCarcinoma 

Overall rate 2/50 (4%) 3/50 (6%) 2/50 (4%) 2/50 (4%) 

Adjusted rate 9.3% 14.3% 6.7% 6.7% 

Terminal rate 1/20 (5 %) 1/14 (7%) 1/22 (5%) 1/28 (4%) 

First incidence (days) 720 660 596 686 

Life table test P=0.388N P=O.371 P=0.669N P=0.603N 

Logistic regression test P=0.506N P=O.418 P=0.687N P=0.661N 

Cochran-Armitage test P=O.S44N 

Fisher exact test P=O.500 P=0.691N P=0.691N 

MammaryGland:Fibroadenoma 

Overall rate 3/50 (6%) 8/50 (16%) 3/50 (6%) 3/50 (6%) 

Adjustedrate . 11.7% 31.7% 11.6% 10.7% 

Terminal rate 1/20 (5%) 3/14 (21 96) 2/22 (9%) 3/28 (1 1 X) 

First incidence (days) 673 404 646 726 (T) 

Life table test P=0.142N P=O.O59 P=0.644N P=0.532N 

Logistic regression test P=0.276N P=O.110 P=0.654N P=0.623N 

Cochran-Armitage test P=0.276N 

Fisher exact test P=O.100 P=0.661N P=0.661N 

PancreaticIslets:Adenoma 

Overall rate 3/50 (6%) 2/50 (4%) 1/50 (2%) 0150 (0%) 

Adjusted rate 15.0% 10.9% 4.5% 0.0% 

Terminal rate 3/20 (15%) 1/14 (7%) 1/22 (5%) 0/28 (0%) 

-e First incidence (days) 726 (T) 678 726 (T) 

Life table test P=0.059N P=0.643N P=0.268N P=0.067N 

Logistic regression test P=0.076N P=0.622N P=0.268N P=0.067N 

Cochran-Armitage test P=0.114N 

Fisher exact test P=O.5OON P=0.309N P=0.121N 

PancreaticIslets:Adenoma or Carcinoma 

Overall rate 3/50 (6%) 3/50 (6%) 1/50 (2%) 0150 (0%) 

Adjusted rate 15.0% 17.7% 4.5% 0.0% 

Terminal rate 3/20 (15%) 2/14 (14%) 1/22 (5 %) 0128 (0%) 

First incidence (days) 726 (T) 678 726 (T) -

Life table test P=0.038N P=O.508 P=0.268N P=0.067N 

Logistic regression test P=O.OSlN P90.530 P=0.268N P=0.067N 

Cochran-Armitage test P=0.084N 

Fisher exact test P=0.661N P=0.309N P=O.l21N 
Lesions 97 in Male Rats 

TABLEA3 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 

PituitaryGland(Pars Distalis): Adenoma 

Overall rate 19/49 (39%) 

Adjusted rate 57.6% 

Terminal rate 7/19 (37%) 

First incidence (days) 471 

Life table test P=0.009N 

Logistic regression test P=0.033N 

Cochran-Armitage test P=0.034N 

Fisher exact test 

PreputialGland:Adenoma 

Overall rate 3/50 (6%) 

Adjusted rate 11.1% 

Terminal rate 1/20 (5 %) 

First incidence (days) 47 1 

Life table test P=0.425N 

Logistic regression test P=0.572N 

Cochran-Armitage test P=0.582N 

Fisher exact test 

PreputialGland:Adenoma or Carcinoma 

Overall rate 3/50 (6%) 

Adjusted rate 11.1% 

Terminal rate 1/20 (5 %) 

First incidence (days) 47 1 

Life table test P=0.351N 

Logistic regression test P=0.492N 

Cochran-Armitage test P=0.492N 

Fisher exact test 

Skin:Fibroma 

Overall rate 5/50 (10%) 

Adjusted rate 15.7% 

Terminal rate 1/20 (5 %) 

First incidence (days) 649 

Life table test P=0.278N 

Logistic regression test P=0.410N 

Cochran-Armitage test P=0.440N 

Fisher exact test 

Skin: SquamousCellPapilloma 

Overall rate 0/50 (0%) 

Adjusted rate 0.0% 

Terminal rate 0/20 (0%) 

First incidence (days) -

Life table test P=0.516N 

Logistic regression test P=0.617N 

Cochran-Armitage test P=0.636N 

Fisher exact test 

1 mgkg 

17/49 (35%) 

52.1% 

2/14 (14%) 

446 

P=O.445 

P=0.450N 

P=0.417N 

1/50 (2%) 

7.1% 

1/14 (7%) 

726 (T) 

P=0.406N 

P=0.311N 

P=0.309N 

2/50 (4%) 

9.1% 

1/14 (7%) 

492 

P=0.594N 

P=0.483N 

P=0.500N 

3/50 (6%) 

13.0% 

1/14 (7%) 

656 

P=0.454N 

P=0.387N 

P=0.357N 

3/50 (6%) 

16.6% 

2/14 (14%) 

589 

P=O.O78 

P=O.106 

P=O.121 

5 mgkg 

13/50 (26%) 

42.7% 

6/22 (27%) 

63 1 

P=0.132N 

P=0.108N 

P=0.126N 

3/49 (6%) 

9.5% 

0121 (0%) 

649 

P=O.658 

P=O.651 

P=O.651 

3/49 (6%) 

9.5% 

0/21 (0%) 

649 

P=O.658 

P=O.651 

P=O.651 

3\50 (6%) 

11.8% 

2/22 (9%) 

656 

P=0.359N 

P=0.346N 

P=0.357N 

0150 (0%) 

0.0% 

0122 (0%) -

-

-

-

25 mgkg 

10150 (20%) 

27.1% 

5/28 (18%) 

35 1 

P=0.017N 

P=0.033N 

P=0.033N 

2/50 (4%) 

7.1% 

2/28 (7%) 

726 (T) 

P=0.391N 

P=0.501N 

P=0.500N 

2/50 (4%) 

7.1 % 

2/28 (7%) 

726 (T) 

P=0.391N 

P=0.501N 

P=0.500N 

3/50 (6%) 

10.0% 

2/28 (7%) 

685 

P=0.267N 

P=0.352N 

P=0.357N 

1/50 (2%) 

3.6% 

1/28 (4%) 

726 (T) 

P=O.567 

P=O.567 

P=O.500 
98 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA3 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Skin:Keratoacanthoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

Skin:SquamousCellPapilloma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

Vehicle Control 

4/50 (8%) 

18.7% 

3/20 (15%) 

713 

P=0.161N 

P=0.224N 

P=0.330N 

or Squamous CellCarcinoma 

1/50 (2%) 

5.0% 

1/20 (5%) 

726 (T) 

P=0.352N 

P=0.452N 

P=0.487N 

1 mgk! 5 mgh 

3/50 (6%) 3/50 (6%) 

18.6% 12.0% 

2/14 (14%) 2/22 (9%) 

702 666 

P=O.627 P=0.459N 

P=O.651 P=0.499N 

P=0.500N P=0.500N 

3/50 (6%) 01.50 (0%) 

16.6% 0.0% 

2/14 (14%) 0/22 (0%) 

589 -

P=O.206 P=0.481N 

P=O.261 P=0.481N 

P=O.309 P=0.500N 

Skin:SquamousCellPapilloma,Keratoacanthoma, Basal Cell Adenoma, or Squamous Cell Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

Testes:Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

ThyroidGland(C-cell):Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

4/50 (8%) 5/50 (10%) 3/50 (6%) 

18.7% 27.4% 12.0% 

3/20 (15%) 3/14 (21%) 2/22 (9%) 

713 589 666 

P=0.304N P=O.306 P=0.459N 

P=0.449N P=O.378 P=0.499N 

P=0.561N 

P=O.500 P=0.500N 

43/50 (86%) 42/50 (84%) 46/50 (92%) 

100.0% 100.0% 100.0% 

20/20 (100%) 14/14 (100%) 22/22 (100%) 

443 404 565 

P=0.022N P=O.128 P=O.499 

P=O.502 P=O.617 P=O.342 

P=O.452 

P=0.500N P=O.262 

1/50 (2%) 4/50 (8%) 3/50 (6%) 

3.1% 16.1% 9.9% 

0120 (0%) 1/14 (7%) 0122 (0%) 

25 mgkg 

2/50 (4%) 

6.8% 

1/28 (4%) 

692 

P=0.220N 

P=0.274N 

P=0.339N 

1/50 (2%) 

3.6% 

1/28 (4%) 

726 (T) 

P=0.686N 

P=0.686N 

P=0.753N 

4/50 (8%) 

13.7% 

3/28 (1 1 %) 

692 

P=0.475N 

P=0.559N 

P=0.643N 

44/50 (88%) 

100.0% 

28/28 (100%) 

504 

P=O.llON 

P=O.408 

P=O.500 

1/50 (2%) 

3.6% 

1/28 (4%) 

First incidence (days) 504 663 646 726 (T) 

Life table test P=0.208N P=O.141 P=O.307 P=0.706N 

Logistic regression test P=0.296N P=O.179 P=O.308 P=0.759N 

Fisher exact test 

Cochran-Armitage test P=0.298N 

P=O.181 P=O.309 P=0.753N 
Lesions in Male Rats 99 

TABLEA3 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 

ThyroidGland(C-cell):Carcinoma 

Overall rate 1/50 (2%) 

Adjusted rate 5.0% 

Terminal rate 1/20 (5%) 

First incidence (days) 726 (T) 

Life table test P=0.253N 

Logistic regression test P=0.253N 

Cochran-Armitage test P=0.382N 

Fisher exact test 

ThyroidGland(C-cell):Adenoma or Carcinoma 

Overall rate 2/50 (4%) 

First incidence (days) 

Terminal rate 

Adjusted rate 

663 

1 /20 (5X) 

8.0% 

Life table test P=0.094N 

Cochran-Armitage test 

Logistic regression test 

P=0.184N 

P=0.169N 

Fisher exact test 

ThyroidGland(FollicularCell):Adenoma 

Overall rate 3/50 (6%) 

First incidence (days) 

Terminal rate 

Adjusted rate 

430 

1/20 (5X) 

9.3% 

Life table test P=0.081N 

Logistic regression test P=0.114N 

Cochran-Armitage test P=0.114N 

Fisher exact test 

ThyroidGland(FollicularCell):Adenoma or Carcinoma 

Overall rate 4/50 (8%) 

Adjusted rate 14.0% 

Terminal rate 2/20 (10%) 

First incidence (days) 430 

Life table test P=0.050N 

Logistic regression test P=0.078N 

Cochran-Armitage test P=0.079N 

Fisher exact test 

All Organs:Mesothelioma NOS 

Overall rate 1/50 (2%) 

Adjusted rate 3.7% 

Terminal rate 0/20(0%) 

First incidence (days) 698 

Life table test P=O.422 

Logistic regression test P=O.298 

Cochran-Armitage test P=O.292 

Fisher exact test 

1 mgm 

Ob0 (0%) 

0.0% 

0114 (0%) -

P=0.571N 

P=0.571N 

P=0.500N 

4/50 (8 46) 

16.1X 

1/14 (7%) 

504 

P=O.258 

P=O.325 

P=O.339 

2/50 (4%) 

10.9% 

1/14 (7%) 

678 

P=0.598N 

P=0.492N 

P=0.500N 

2/50 (4%) 

10.9% 

1/14 (7%) 

678 

P=0.454N 

P=0.340N 

P=0.339N 

1/50 (2%) 

3.6% 

0/14 (0%) 

666 

P=O.715 

P=O.746 

P=0.753N 

5 mgflrg 

3/50 (6%) 

13.6% 

3/22 (14%) 

726 (T) 

P=O.337 

P=O.337 

P=O.309 

6/50 (12%) 

22.2% 

3/22 (14%) 

646 

P=O.155 

P=O.140 

P=O.134 

1/50 (2%) 

4.2% 

0122 (0%) 

713 

P=0.303N 

P=0.314N 

P=0.309N 

1/50 (2%) 

4.2% 

0122 (0%) 

713 

P=O. 175N 

P=0.183N 

P=0.181N 

4/50 (8%) 

10.9% 

1/22 (5 X) 

565 

P=O.201 

P=O.174 

P=O.181 

25 mgikg 

0150 (0%) 

0.0% 

0128 (0%) -

P=0.433N 

P=0.433N 

P=0.500N 

1/50 (2%) 

3.6% 

1/28 (4%) 

726 (T) 

P=0.406N 

P=0.482N 

P=0.500N 

0150 (0%) 

0.0% 

0128 (0%) -

P=O.llON 

P=0.106N 

P=0.121N 

0150 (0%) 

0.0% 

0128 (0%) -

P=0.049N 

P=0.060N 

P=0.059N 

3/50 (6%) 

10.7% 

3/28 (11%) 

726 (T) 

P=O.406 

P=O.354 

P=O.309 
100 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA3 

Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Gavage Study 

of Scopolamine' Hydrobromide Trihydrate(continued) 

~~ 

Vehicle Control 1 mgkg 5 mgkg 25 mgkg

AU Organs:MononuclearCellLeukemia 

Overall rate 33/50 (66%) 21/50 (42%) 26/50 (52 %) 24/50 (48 %) 

Adjusted rate 86.0% 73.3% 66.5% 59.3% 

Terminal rate 15/20 (75%) 8/14 (57%) 10122 (45 A) 12/28 (43 %) 

First incidence (days) 443 533 463 504 

Life table test P=0.034N P=0.240N P=0.139N P=0.017N 

Logistic regression test P=0.243N P=0.022N P=0.093N P=0.052N 

Cochran-Armitage test P=0.265N 

Fisher exact test P=0.013N P=O.lllN P=0.053N 

All Organs: Benign Neoplasms 

Overall rate 49/50 (98%) 49/50 (98 %) 48/50 (96%) 49/50 (98 %) 

Adjusted rate 100.0% 100.0% 100.0% 100.0% 

Terminal rate 20/20 (100 %) 14/14 (100%) 22/22 (100%) 28/28 (100%) 

First incidence (days) 430 404 565 144 

Life table test P=0.022N P=O.111 P=0.389N P=O.lllN 

Logistic regression test P=O.129 -f P=0.174N P=O.365 

Cochran-Armitage test P=O.627 

Fisher exact test P=0.753N P=0.500N P=0.753N 

All Organs: Malignant Neoplasms 

Overall rate 39/50 (78%) 27/50 (54%) 31/50 (62%) 28/50 (56%) 

Adjusted rate 94.9% 84.1% 76.1X 64.5% 

Terminal rate 18/20 (90%) 10114 (71 %) 13/22 (59%) 13/28 (46%) 

First incidence (days) 443 492 463 504 

Life table test P=O.OllN P=0.316N P=0.108N P=0.008N 

Logistic regression test P=0.215N P=0.016N P=0.044N P=0.027N 

Cochran-Armitage test P=0.134N 

Fisher exact test P=O.OlON P=0.063N P=0.016N 

All Organs: Benign or MalignantNeoplasms 

Overall rate 49/50 (98 %) 49/50 (98 X) 49/50 (98 %) 49/50 (98 %) 

Adjusted rate 100.0% 100.0% 100.01 100.0% 

Terminal rate 20/20 (100 %) 14/14 (100%) 22/22 (100%) 28/28 (100%) 

First incidence (days) 430 404 463 144 

Life table test P=0.022N P=O.111 P=0.442N P=O.lllN 

Logistic regression test P=O.101 - - P=O.365 

Cochran-Armitage test P=O.691 

Fisher exact test P=0.753N P=0.753N P=0.753N 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animalslnumber of animals examined. Denominator is number of animals examined microscopically for adrenal gland, liver, 

pancreatic islets, pituitary gland, preputial gland, testes, and thyroid gland; for other tissues, denominator is number of animals necropsied. 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence at terminal kill 

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-Annitage 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 

Value of statistic cannot be computed. 
Lesions in Male Rats 101 

TABLEA4a 

Historical Incidence of Pituitary Gland (Pars Distalis) Adenoma 

in Male F344/N Rats Administered Water by Gavagea 

Incidence in Controls 

Overall Historical Incidence 

Total 1161363 (32.0%) 

Standard deviation 7.7 x 

Range 2458043% 

a Data as of 17 June 1994 

TABLEA4b 

Historical Incidence of Mononuclear Cell Leukemia in Male F344/N Rats Administered Water by Gavagea 

Incidence in Controls 

Overall Historical Incidence 

Total 1731367 (47.1 %) 

Standard deviation 9.2% 

Range 34%-56% 

a Data as of 17 June 1994; includes data for lymphocytic, monocytic, and undifferentiated leukemia 
102 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA5 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydratea 

ControlVehicle 1 mgkg 5 mgkg 

Disposition Summary 

Animals initially in study 60 60 60 

15-Month interim evaluation 10 10 10 

Early deaths 

Accidental deaths 2 2 1 

Moribund 23 30 20 

Natural deaths 5 4 7 

Survivors 

Terminal sacrifice 20 14 22 

Animals examined microscopically 60 60 60 

IS-Month Interim Evaluation 

Alimentary System 

Intestine large, colon 

Parasite metazoan 

Intestine large, rectum 

Parasite metazoan 

Liver 

Basophilic focus 

Hepatodiaphragmatic nodule 

Hyperplasia 

Mesentery 

Fat, necrosis 

Pancreas 

Acinus, atrophy 

Cardiovascular System 

Heart 

Myocardium, degeneration 

Endocrine System 

Adrenal cortex 

Hyperplasia 

Pituitary gland 

cyst 

Pars distalis, hyperplasia 

Thyroid gland 

C-cell, hyperplasia 

Follicle, dilatation 

Genital System 

Preputial gland 

Inflammation, suppurative 

Testes 

Atrophy 

Interstitial cell, hyperplasia 

a Number of animals examined microscopically at the site and the number of animals with lesion 

25 mgkg 

60 

10 

1 

17 

4 

28 

60 
Lesions in Male Rats 103 

of Scopolamine Hydrobromide Trihydrate (continued) 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Gavage Study 

TABLEA5 

Vehicle Control 25 mgkg 

Spleen 

HematopoieticSystem 

15-Month Interim Evaluation (continued) 

Thymus (9) 

(10) 

Atrophy 

Fibrosis 

Lung 

Respiratory System 

Interstitium, inflammation 

Alveolar epithelium, hyperplasia 

(10) 

1(10%) 

(10) 

1 (10%) 

(10) 

1(10%) 

(10) 

1 (10%) 

Inflammation. chronic 

Ear 

Special Senses System 

(3)

3 (100%) 

Nephropathy 

Kidney 

Urinary System 

Nervous System 

MusculoskeletalSystem 

IntegumentarySystem 

General Body System 

Systems Examined With No Lesions Observed 

2-Year Study 

AlimentarySystem 

Periesophageal tissue, inflammation, chronic 

Ulcer 

Inflammation, chronic 

Esophagus 

Ulcer 

Parasite metazoan 

Intestine large, colon 

Intestine large, cecum 

Parasite metazoan 

Intestine large, rectum 

Ulcer 

Parasite metazoan 

Ulcer 

Inflammation, chronic 

Intestine small, jejunum 

Inflammation, chronic 

Intestine small, ileum 

1 (2%) 
104 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEA5 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

2-YeUr study (continued) 

Alimentary System (continued) 

Liver 

Basophilic focus 

Clear cell focus 

cyst 

Degeneration, cystic 

Eosinophilic focus 

Fatty change 

Hepatodiaphragmatic nodule 

Inflammation, chronic 

Mineralization 

Mixed cell focus 

Necrosis 

Centrilobular, degeneration 

Portal, degeneration 

Sinusoid, congestion, focal 

Mesentery 

Inflammation, suppurative 

Artery, thrombosis 

Fat, inflammation 

Fat, necrosis 

Lymphatic, cyst 

Pancreas 

Acinus, atrophy 

Arteriole, inflammation, chronic 

Artery, inflammation, chronic 

Stomach, forestomach 

Hyperkeratosis 

Inflammation, chronic 

Ulcer 

Stomach, glandular 

Erosion 

Mineralization 

Ulcer 

Tongue 

Foreign body 

Cardiovascular System 

Heart 

Atrium, thrombosis 

Myocardium, degeneration 

Valve, inflammation, chronic 

Valve, thrombosis 

Endocrine System 

Adrenal cortex 

Hyperplasia 

Adrenal medulla 

Hyperplasia 

Necrosis 

Hyperplasia 

VehicleControl 

(50) 

4 (8%) 

1(2%) 

1 (2%) 

9 (18%) 

2(4%) 

4 (8%) 

2(4%) 

1(2%) 

1(2%) 

1(2%) 

(7) 

1(14%) 

6 (86%) 

1(14%) 

(50) 

18 (36%) 

1(2%) 

(50) 

3 (6%) 

(50) 

1(2%) 

8 (16%) 

(50) 

3 (6%) 

40 (80%) 

1 (2%) 

1(2%) 

3(6%) 

3(6%) 

2(4%) 

3 (6%) 

1(2%) 

1(2%) 

2(4%) 

1(2%) 

(13) 

1 (8%) 

12 (92%) 

(50) 

15 (30%) 

1 (2%) 

(50) 

2(4%) 

43 (86%) 

5 mg/kg 

(50) 

4 (8%) 

1(2%) 

4 (8%) 

2(4%) 

5 (10%) 

6(12%) 

1(2%) 

17 (77%) 

1(2%) 

(50) 

4 (8%) 

(50) 

1 (2%) 

2(4%) 

(50) 

5 (10%) 

40 (80%) 

25 mg/kg 

4 (8%) 

5 (10%) 

2(4%) 

1(2%) 

1(2%) 

1(2%) 

1 (7%) 

12 (86%) 

(50) 

13 (26%) 

(50) 

3(6%) 

34 (68%) 
Lesions in Male Rats 105 

TABLEA5 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

&Year Study (continued) 

Endocrine System (continued) 

Islets, pancreatic 

Parathyroid gland 

Hyperplasia 

Angiectasis 

Pituitary gland (49) (50) 

1 (2%) 

cyst 4 (8%) 5 (10%) 3 (6%) 

Pars distalis, hyperplasia 14 (29%) 17 (35%) 9 (18%) 

Pars intermedia, hyperplasia, tubular 

Thyroid gland (50) 

C-cell, hyperplasia 2 (4%) 

C-cell, infiltration cellular 

Follicle, dilatation 

General Body System 

None 

Genital System 

Preputial gland 

Ectasia 

(50) 

1 (2%) 

(49) 

Fibrosis 1 (2%) 

Hyperplasia 

Inflammation, chronic 

Inflammation, suppurative 

Prostate 

Inflammation 

Inflammation, chronic 

Inflammation, suppurative 

Seminal vesicle 

Fibrosis 

Testes 

Germinal epithelium, atrophy 

HematopoieticSystem 

Bone marrow 

Fibrosis 

Hyperplasia 

Lymph node 

Lumbar, angiectasis 

Mediastinal, angiectasis 1 (13%) 

Mediastinal, pigmentation, hemosiderin 1 (20%) 

Renal, pigmentation, hemosiderin 

Lymph node, mandibular 

Hemorrhage 

Infiltration cellular, plasma cell 

Lymph node, mesenteric 

Congestion 

Fibrosis 

Inflammation, suppurative 1 (2%) 

5 (10%) 

18 (36%) 

1 (2%) 

(50) 

3 (6%) 

1 (2%) 

(50) 

2 (4%) 

1(2%) 

(50) 

(50) 

3 (6%) 

(50) 

1 (2%) 

(9) 

1 (11%) 

1(11%) 

(49) 
106 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEA5 

Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

VehicleControl 1 mgkg . 25 mgkg 

HematopoieticSystem (continued) 

2-YeUr study (continued) 

Spleen 

Fibrosis 

Hematopoietic cell proliferation 

Hyperplasia, lymphoid 

Necrosis 

Thymus 

Atrophy 

Congestion 

IntegumentarySystem 

Mammary gland 

Hyperplasia, cystic 

Lymphatic, ectasia 

Skin 

Hyperkeratosis 

Hyperplasia, basal cell 

Inflammation, acute 

Ulcer 

Dermis, fibrosis, focal 

Hair follicle, atrophy, focal 

Subcutaneous tissue, edema 

MusculoskeletalSystem 

Bone 

Fibrous osteodystrophy 

NervousSystem 

Brain 

Hypothalamus, hemorrhage 

Medulla, gliosis 

Ventricle, hydrocephalus 

RespiratorySystem 

Lung 

Ectopic tissue 

Foreign body 

Inflammation, chronic 1 (2%) 1 (2%) 

Inflammation, subacute 

Inflammation, granulomatous 

1 (2%) 3 (6%) 

1 (2%) 

Alveolar epithelium, hyperplasia 1(2%) 

Alveolus, edema 1 (2%) 

Pleura, inflammation, chronic 

Nose 

Foreign body 

Inflammation, suppurative 
Lesions in Male Rats 107 

TABLEA5 

Summary of the Incidence of Nonneoplastic Lesions in MaleRats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

%Year Study (continued) 

Special Senses System 

Far 

External ear, hyperkeratosis 

External ear, inflammation, chronic 

Eye

Degeneration 

Lens, cataract 

Harderian gland 

Inflammation, suppurative 

Urinary System 

Kidney 

cyst 

Nephropathy 

Renal tubule, pigmentation, hemosiderin 

Transitional epithelium, hyperplasia 

Urinary bladder 

Inflammation, acute 

Inflammation, suppurative 

Ulcer 

Transitional epithelium, hyperplasia 

Vehicle Control 1 mgQ 5 mag 25 mglkg 

(3) 

2 (67%) 

1 (50%) 

(1) (1) (1) 

1 (100%) 

1 (100%) 1 (100%) 

(1) (1) 

1 (100%) 

(50) 

48 (96%) 

(50) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 
10s Scopolamine Hydrobromide Trihydrate, NTP TR 445 
109 

APPENDIX B 

SUMMARY OF LESIONS IN FEMALE RATS 

IN THE 2-YEAR GAVAGE STUDY 

OF SCOPOLAMINE HYDROBROMIDE TRIHYDRATE 

TABLE B1 Summary of the Incidence of Neoplasms in Female Rats 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........... 110 

TABLEB2 Individual Animal Tumor Pathology of Female Rats 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........... 114 

TABLE B3 Statistical Analysis of Primary Neoplasms in Female Rats 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ............ 130 

TABLE B4a Historical Incidence of Pituitary Gland (Pars Distalis) Adenoma 

in Female F344/N Rats Administered Water by Gavage ..................... 133 

TABLEB4b Historical Incidence of Mononuclear Cell Leukemia 

in Female F344/NRats Administered Water by Gavage ..................... 133 

TABLEB4c Historical Incidence of Thyroid Gland (Follicular Cell) Adenoma 

in Female F344/N Rats Administered Water by Gavage ..................... 133 

TABLEB5 Summary of the Incidence of Nonneoplastic Lesions in Female Rats 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........... 134 
110 Scopolamine Hydrobromide Trihydrate, NTP Tk 445 

Early deaths 

IS-Month interim evaluation 

Animals initially in study 

Disposition Summary 

Natural deaths 

Moribund 

Accidental deaths 

Terminal sacrifice 

Survivors 

10 

60 

3 

13 

34 

60 

15 

10 

18 

17 

10 

60 

16 

6 

2 

26 

10 

60 

16 

7 

5 

22 

Animals examined microscopically 60 60 60 60 

Pituitary gland 

Endocrine System 

15-Month Interim Evaluation 

Pars distalis, adenoma 

(10) 

1 (10%) 

Polyp stromal 

Uterus 

Genital System 

Fibroadenoma 

Mammary gland 

integumentary System 

(9) 

1 (11%) 

Urinary System 

Special Senses System 

Respiratory System 

Nervous System 

Musculoskeletal System 

Hematopoietic System 

General Body System 

Cardiovascular System 

Alimentary System. 

Systems Examined With No Neoplasms Observed 
Lesions in FemaleRats 111 

TABLEB1 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

2-Year Study 

Liver 

AlimentarySystem 

Tongue 

Hepatocellular adenoma 

(50) 

1 (2%) 

(60) 

Squamous cell carcinoma 

Tooth 

Gingiva, squamous cell carcinoma 

Cardiovascular System 

Heart 

Schwannoma NOS 

EndocrineSystem 

Adrenal cortex 

Adenoma 

Pheochromocytoma benign 

Adrenal medulla (50) 

1(2%) 

Bilateral, pheochromocytoma malignant 

Islets, pancreatic 

Adenoma 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Bilateral, C-cell, adenoma 

Ccell, adenoma 1 (2%) 2 (4%) 1 (2%) 

C-cell, carcinoma 1 (2%) 

Follicular cell. adenoma 3 (6%) 

None 

General Body System 

Genital System 

Clitoral gland 

Adenoma 

Bilateral, adenoma 

Uterus 

Adenoma 

Polyp stromal 

Schwannoma NOS 
112 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEB1 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Control Vehicle 1 mgk 5 mgm 25 mgkg

2-Year Study (continued) 

HematopoieticSystem 

Bone marrow (50)

Pheochromocytoma malignant, metastatic, 

adrenal medulla 1 (2%) 

Lymph node (2)

Lymph node, mandibular (50) (50)

Histiocytic sarcoma 

Lymph node, mesenteric (50)

Histiocytic sarcoma 

Spleen (50)

Histiocytic sarcoma 

Pheochromocytoma malignant, metastatic, 

adrenal medulla 

Thymus (48)

Thymoma malignant 

IntegumentarySystem 

Mammary gland 

Adenoma 

Carcinoma 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Keratoacanthoma 

Sarcoma 

Sebaceous gland, adenoma 

Musculoskeletal System 

Skeletal muscle (1)

Rhabdomyosarcoma 1 (100%) 

NervousSystem 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung (50) (50)

Pheochromocytoma malignant, metastatic, 

adrenal medulla 1 (2%) 

Special Senses System 

EY e 
Lesions in Female Rats 113 

TABLEB1 

Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle Control 1 mgm 

&Year Study (continued) 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organsb (50) (60)

Histiocytic sarcoma 1 (2%) 

Leukemia mononuclear 20(40%) 6(10%) 

Neoplasm Summary 

Total animals with primary neoplasms' 

15-Month interim evaluation 2 

2-Year study 48 23 

Total primary neoplasms 

15-Month interim evaluation 2 

2-Year study 92 37 

Total animals with benign neoplasms 

15-Month interim evaluation 2 

2-Year study 37 19 

Total benign neoplasms 

15-Month interim evaluation 2 

2-Year study 66 28 

Total animals with malignant neoplasms 

2-Year study 23 9 

Total malignant neoplasms 

2-Year study 24 9 

Total animals with metastatic neoplasms 

2-Year study 

Total metastatic neoplasms 

2-Year study 

Total animals with uncertain neoplasmsbenign or malignant 

2-Year study 2 

Total uncertain neoplasms 

2-Year study 2 

a 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 

5 mgk 

(50) 

1 (2%) 

13(26%) 

1 

39 

1 

64 

1 

32 

1 

46 

16 

18 

25 mgkg 

(50) 

4 (8%) 

2 

31 

2 

43 

2 

28 

2 

37 

6 

6 

I 

3 
114 Scopolamine Hydrobrimide Trihydrate,NTP TR 445 

TABLEB2 

Individual AnimalTumor Pathology of Female Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl 

Number of DaysonStudy 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular adenoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Schwannoma NOS 

EndocrineSystem 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Bilateral, C-cell, adenoma 

C-cell, adenoma 

C-cell, carcinoma 

Follicular cell, adenoma 

General Body System 

None 

GenitalSystem 

Clitoral gland 

Adenoma 

Ovary 

UteNS 

Polyp stromal 

Schwannoma NOS 

Vagina 

+: Tissue examined microscopically 

A: Autolysis precludes examination 

3444555666666666777777777 

7366668112566789222222222 

4713222117803153666666666 

2222222222222222222222222 

6577684995944858445566677 

7329287174395570232905945 

......................... 

......................... 

......................... 

+++++++A+++++++++++++++++ 

......................... 

+++++++A+++++++++++++++++ 

......................... 

......................... X 

.......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

.......................... 

......................... X ......................... 

++++M++MM++MM++++++++++++ 

......................... xx xx X x x xxx ......................... 

......................... X 

......................... 

......................... xxx 

X 

+ + 

M: Missing tissue X: Lesion present 

I: Insufficient tissue Blank: Not examined 
Lesions in Female Rats 115 

TABLEB2 

Individual Animal Tumor Pathologyof Female Rats in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

Number of DaysonStudy 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular adenoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

CardiovascularSystem 

Blood vessel 

Heart 

Schwannoma NOS 

EndocrineSystem 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma benign 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Bilateral, C-cell, adenoma 

C-cell, adenoma 

C-cell, carcinoma 

Follicular cell, adenoma 

GeneralBodySystem 

None 

GenitalSystem 

Clitoral gland 

Adenoma 

Ovary 

Uterus 

Polyp stromal 

Schwannoma NOS 

Vagina 

~~~ ~ ~~ ~~ ~~ ~~ ~~~ 

1771177177777777777777777 

2222222222222222222222222 

6666666666688888888888888 

2222222222322222222222222 Total 

7888888999044455566611999 Tissues1 

7123619256046801514601048 Tumors 

++++++++++++++++++++++M++ 49 

......................... 50 

......................... 50 

.......................... 49 

......................... 50 

......................... 49 

......................... 50 

......................... 50 

1 

+ 1 

......................... 50 

.......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

X 1 

......................... 50 

......................... 50 

1 

......................... 50 

+++++++++++++M+MM++++M+++ 41 

......................... 50 

X x x x xxx x X 20 

.......................... 50 

X 1 

X X 2 

xx 2 

X 1 

++++++++++++++++++++M++++ 49 

X X 3 

......................... 50 

......................... 50 

X X X 6 

1 

2 
116 Scopolamine Hydrobrimide Trihydrate, NTP TR 445 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: Vehicle Control (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Blood 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

Integumentary System 

Mammary gland 

Carcinoma 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Keratoacanthoma 

Sarcoma 

Sebaceous gland, adenoma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Nose 

Trachea 

Special Senses System 

Ear 

EYe 

Urinary System 

Kidney 

Urinary bladder 

~~~~~~ ~ 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

344455566666,6666111111111 

1366668112566189222222222 

4113222111803153666666666 

2222222222222222222222222 

6577684995944858445566677 

7329281114395570232905945 

......................... + + ......................... 

......................... 

......................... 

......................... 

......................... 

x xxxx X X X 

X X x xxx .......................... 

X 

......................... 

.......................... + + + + + 

+ + 

......................... 

......................... 

......................... 

+ 

+ + 

......................... 

......................... 

......................... x xx x x x x xxxxx X 
Lesions in Female Rats 117 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromide Trihydrate: VehicleControl (continued) 

Number of Days on Study 

Carcass ID Number 

~~ ~ 

HematopoieticSystem 

Blood 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

IntegumentarySystem 

Mammary gland 

Carcinoma 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Keratoacanthoma 

Sarcoma 

Sebaceous gland, adenoma 

Musculoskeletal System 

Bone 

~ ~~~ 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Nose 

Trachea 

Special Senses System 

Ear 

EYe ~~ 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

7777777777777777777777777 

2222222222222222222222222 

6666666666688888888888888 

2222222222322222222222222 Total 

7888888999044455566617999 Tissues1 

7123679256046801514601048 Tumors 

+ 1 

......................... 50 

2 

......................... 50 

......................... 50 

......................... 50 

M++++++++++++++++++M+++++ 48 

......................... 50 

X 1 

X x x xx xxxxxx x 20 

X X X 9 

......................... 50 

X 1 

X 1 

1 

......................... 50 

......................... 50 

5 

2 

......................... 50 

......................... 50 

......................... 50 

+ 2 

2 

~ ~~ 

......................... 50 

......................... 50 

......................... 50 

X X x x x x X 20 
118 Scopolamine Hydrobrimide Trihydrate, NTP TR 445 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 1 mg/kg 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

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 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

C-cell, carcinoma 

General Body System 

None 

Genital System 

Clitoral gland 

Adenoma 

Ovary 

Uterus 

Polyp stromal 

HematopoieticSystem 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Histiocytic sarcoma 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Histiocytic sarcoma 

Thymus 

122222222222222222233333444445 

700000000000000003955777026661 

844444444444444449978799823377 

333333333333333333333333333333 

111111111122222220332404340225 

012345678901234567169014919781 

.............................. 

.............................. 

.............................. 

.............................. 

.............................. 

+++M++++++++++++++++++AM++++++ 

+++A+++++++++++++++.+++AM++++++ 

.............................. + + .............................. 

.............................. 

.............................. 

.............................. ~ ~ ~~ ~ ~~~~ ~~~ 

.............................. 

.............................. 

.............................. 

.............................. 

.............................. 

++++M++++++++++++++++++M++++++ 

.............................. X X 

.............................. 

.............................. 

............................... 

.............................. X 

.............................. 

.............................. 

++++++++++M+++++++++++++++++++ 

.............................. 

.............................. 
Lesionsin Female Rats 119 

TABLEB2 

Individual Animal Tumor Pathologyof Female Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: 1mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

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 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Parathyroid gland 

Pituitaly gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

C-cell, carcinoma 

General Body System 

None 

Genital System 

Clitoral gland 

Adenoma 

Ovary 

Uterus 

Polyp stromal 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Histiocytic sarcoma 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Histiocytic sarcoma 

Thymus 

55555556666667777777777777777~ 

144667802223322222222222222222 

914222134885566688888888888888 

333333333333333333333333333333 Total 

554034535343545500000333444556 Tissues1 

769617008455582423458238236390 Tumors 

+++++M+++++++++++++++++++M++++ 58 

+++++++M++++++++++++++++++++++ 59 

+++++++M++++++++++++++++++++++ 59 

+++++++M++++++++++++++++++++++ 59 

.............................. 60 

+++++++M++++++++++++++++++++++ 56 

+++++++M++++++++++++++++++++++ 56 

.............................. 60 

+ 3 

.............................. 60 

.............................. 60 

.............................. 60 

.............................. 60 

.............................. 60 

.............................. 60 

.............................. 60 

.............................. 60 

.............................. 60 

++++++++++++++M++M++++++MM++++ 54 

.............................. 60 

x x xxx xxxxx X 13 

.............................. 60 

X 1 

X 1 

.............................. 60 

X X 2 

.............................. 60 

.............................. 60 

1 

.............................. 60 

+ + 2 

.............................. 60 

X 1 

.............................. 59 

X 1 

.............................. 60 

X 1 

++++++++++++++++++++++++M+++++ 59 
120 Scopolamine Hydrobrimide Trihydrate, NTP TR 445 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromide Trihydrate: 1mg/kg (continued) 

Number of DaysonStudy 

Carcass ID Number 

Integumentary System 

Mammarygland 

Carcinoma 

Fibroadenoma 

Fibroadenoma. multiple 

Skin 

Keratoacanthoma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheralnerve 

Spinal cord 

Respiratory System 

Lung 

Nose 

Trachea 

Special Senses System 

Ear 

Eye 

Urinary System 

Kidney 

Urinarybladder 

Systemic Lesions 

Multiple organs 

Histiocytic sarcoma 

Leukemia mononuclear 

122222222222222222233333444445 

700000000000000003955777026661 

844-444444444444449918799823311 

333333333333333333333333333333 

111111111122222220332404340225 

012345678901234567169014919781 

.............................. 

.............................. 

.............................. 

.............................. 

~~ ~~ ~~~ 

.............................. 

.............................. 

.............................. 

.............................. 

.............................. 

.............................. 
Lesions in Female Rats 121 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromide Trihydrate: 1 mg/kg (continued) 

Number of DaysStudy on 

Carcass ID Number 

IntegumentarySystem 

Mammary gland 

Carcinoma 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Keratoacanthoma 

MusculoskeletalSystem 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Nose 

Trachea 

Special Senses System 

Ear 

EY e 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Histiocytic sarcoma 

Leukemia mononuclear 

555555566666677777777777177777 

144667802223322222222222222222 

914222134885566688888888888888 

333333333333333333333333333333 Total 

554034535343545500000333444556 Tissues/ 

769677008455582423458238236390 Tumors 

.............................. 60 

X 1 

X X X xxx xx 8 

X X 2 

.............................. 60 

X 1 

.............................. 60 

.............................. 60 

+++++ + 6 

+++ + 4 

.............................. 60 

.............................. 60 

.............................. 60 

+ 1 

+ + + 3 

.............................. 60 

.............................. 60 

.............................. 60 

X 1 

X X xx X X 6 
122 Scopolamine Hydrobrimide Trihydrate,NTP .TR 445 

TABLEB2 

Individual Animal Tumor Pathologyof Female Rats in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate: 5 mg/kg 

Number of DaysonStudy 

Carcass ID Number 

AlimentarySystem 

Esophagus 

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 

Tongue 

Squamous cell carcinoma 

Tooth 

Gingiva, squamous cell carcinoma 

Cardiovascular System 

Blood vessel 

Heart 

EndocrineSystem 

Adrenal cortex 

Adenoma 

Adrenal medulla 

Pheochromocytoma benign 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

General Body System 

None 

Genital System 

Clitoral gland 

Adenoma 

Bilateral, adenoma 

Ovary 

uterus 

Polyp stromal 

Vagina 

HematopoieticSystem 

Blood 

Bone marrow 

2334445556666666666777777 

3270674450223345589000002 

7534717750221252307007896 

3344333343344443443334333 

790098671670010812'7880686 

8121893921586192401487502 

......................... 

AA++++++++A++++++++++++++ 

AA++++++++A++++++++++++++ 

AA++++++++A++++++++++++++ 

.......................... 

AA++++++++A++++++++++++++ 

AA+++++++++++++++++++++++ 

......................... + + ......................... 

......................... 

......................... 

......................... 

+ 

X 

......................... 

......................... 

......................... X ......................... 

......................... 

++++++++++M++++MM+++M+++M 

......................... x xxx x x xxx ......................... 

......................... X 

......................... 

......................... X 

+ 

......................... 
Lesions in Female Rats 123 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

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 

Tongue 

Squamous cell carcinoma 

Tooth 

Gingiva, squamous cell carcinoma 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adenoma 

Adrenal medulla 

Pheochromocytoma benign 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

C-cell, adenoma 

General Body System 

None 

Genital System 

Clitoral gland 

Adenoma 

Bilateral,adenoma 

Ovary 

Uterus 

Polyp stromal 

Vagina 

HematopoieticSystem 

Blood 

Bone marrow 

7777777777777777777777777 

2222222222222222222222222 

6666666666688888888888888 

3333344444433333333333444 Total 

6789900111166677889999011 Tissues/ 

43627050367789075735,69358 Tumors 

......................... 50 

......................... 47 

......................... 47 

......................... 47 

......................... 50 

......................... 47 

......................... 48 

......................... 50 

+ 3 

......................... 50 

......................... 50 

........................... 50 

......................... 50 

+ 1 

X 1 

1 

1 

......................... 50 

......................... 50 

......................... 50 

X 2 

......................... 50 

X 1 

......................... 49 

++M+MM+++++++++++++M+++++ 41 

......................... 50 

X X xx X 14 

......................... 50 

X X 2 

++++++M++++++++++++++M+++ 48 

X 2 

X 1 

......................... 50 

......................... 50 

x x X 4 

1 

+ 1 

......................... 50 
124 Scopolamine Hydrobrimide Trihydrate, NTP TR 445 

TABLE B2 

Individual Animal Tumor Pathologyof Female Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: 5 mg/kg (continued) 

2334445556666666666777777 

Number of Days on Study 3270674450223345589000002 

7534717750221252307007896 

3344333343344443443334333 

Carcass ID Number 79.00986716700108127880686 

8121893921586192401487502 

Hematopoietic System (continued) 

Lymph node + 

Lymph node, mandibular ......................... Lymph node, mesenteric ......................... Spleen ......................... Histiocytic sarcoma 

Thymus ......................... Thymoma malignant X ~~ ~ ~______ ~~ ~~ ~ ~~ ~ ~~~~_______ ~ ~ ~~~_____ 

Integumentary System 

Mammary gland ......................... Adenoma X 

Carcinoma 

Fibroadenoma X X X X 

Fibroadenoma, multiple X 

Skin ......................... ~~ ~ ~ ~_____ 

Musculoskeletal System 

Bone ......................... 

Nervous System 

Brain ......................... Peripheral nerve + ++ 

Spinal cord + ++ 

Respiratory System 

Lung ......................... Nose ......................... Trachea ......................... 

+ + ~ ~~ ~______ ~~ ~ ~ ~ ~~~ ~ ~~~ ~ 

Urinary System 

Kidney ......................... Urinary bladder ++++++++++M++++++++++++++ 

Systemic Lesions 

Multiple organs ......................... Histiocytic sarcoma 

Leukemia mononuclear X xx x x X 
Lesions in Female Rats 125 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System (continued) 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Histiocytic sarcoma 

Thymus 

Thymoma malignant 

IntegumentarySystem 

Mammary gland 

Adenoma 

Carcinoma 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Nose 

Trachea 

Special Senses System 

EYe 

Urinary System 

Kidney 

Urinaly bladder 

Systemic Lesions 

Multiple organs 

Histiocytic sarcoma 

Leukemia mononuclear 

1111111111111111111111111 

2222222222222222222222222 

6666666666688888888888888 

3333344444433333333333444 Total 

6189900111166611889999011 Tissues/ 

4362105036118901513569358 Tumors 

+ 2 

......................... 50 

......................... 50 

......................... 50 

X 1 

......................... 50 

1 

......................... 50 

1 

X 1 

xxx X X x x x x X 14 

X X xx 5 

......................... 50 

......................... 50 

......................... 50 

+ + 5 

+ + 5 

......................... 50 

......................... 50 

......................... 50 

2 

......................... 50 

......................... 49 

......................... 50 

X 1 

xx xx X xx 13 
126 Scopolamine Hydrobrimide Trihydrate, NTP TR 445 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/lig .. 

0011223333333344445555566 

Number of Days on Study 4579593334799978990277900 

1880412266212435892006037 

4444444444444444444444444 

Carcass ID Number 7654254725674456582544233 

5145428390082353903647153 

Alimentary System 

Esophagus ......................... Intestine large, colon +++++A++A++++++++++++++++ 

Intestine large, rectum +M+++A++A++++++++++++++++ 

Intestine large, cecum +++++A++A++++++++++++++++ 

Intestine small, duodenum +++++A+++++++++++++++++++ 

Intestine small, jejunum +++++A++A+A++++++++++++++ 

Intestine small, ileum +M+++A++A+A++A++++++M++++ 

Liver ......................... Mesentery + 

Pancreas ......................... Salivary glands ......................... Stomach, forestomach +M+++A+++++++++++++++++++ 

Stomach, glandular +++++A+++++++++++++++++++ 

Cardiovascular System 

Blood vessel ......................... Heart ......................... ~~~~ ~~ 

Endocrine System 

Adrenal cortex .......................... Adenoma 

Adrenal medulla ......................... Pheochromocytoma benign X 

Bilateral, pheochromocytoma malignant 

Islets, pancreatic ........................... Adenoma 

Parathyroid gland +M++++++++++++++++++++++M 

Pituitary gland 

Pars distalis, adenoma 

......................... xx X 

Thyroid gland ......................... C-cell, adenoma 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Clitoral gland ......................... Adenoma 

Ovary .......................... Uterus ......................... Adenoma 

Polyp stromal X X 

Hematopoietic System 

Bone marrow ......................... Pheochromocytoma malignant, 

metastatic, adrenal medulla . ,. 
Lesions in FemaleRats 127 

TABLEB2 

Individual Animal Tumor Pathologyof Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

6667777777777777777777777 

Number of Days on Study 2462222222222222222222222 

9736666666688888888888888 

4444444444444444444444444 

Carcass ID Number 6332335667722333445566677 

8085198452478467093726907 

AlimentarySystem 

Esophagus ......................... Intestine large, colon +A+++++++++++++++++++++++ 

Intestine large, rectum ......................... Intestine large, cecum ......................... Intestine small, duodenum ......................... Intestine small, jejunum +A+++++++++++++++++++++++ 

Intestine small, ileum +A+++++++++++++++++++++++ 

Liver ......................... Mesentery + 

Pancreas ......................... Salivary glands ......................... Stomach, forestomach ......................... Stomach, glandular ......................... 

Cardiovascular System 

Blood vessel ++++++++++++++++M++++++++ 

Heart ......................... 

Endocrine System 

Adrenal cortex ......................... Adenoma X 

Adrenal medulla ......................... Pheochromocytoma benign 

Bilateral, pheochromocytoma malignant X 

Islets, pancreatic ......................... Adenoma X 

Parathyroid gland ++++M++++++++M+++++++++++ 

Pituitary gland ......................... Pars distalis, adenoma x x xx X xx 

Thyroid gland ......................... C-cell, adenoma X 

Follicular cell, adenoma X xx 

General Body System 

None 

Genital System 

Clitoral gland ......................... Adenoma X 

Ovary ......................... Uterus ......................... Adenoma X 

Polyp stromal X 

HematopoieticSystem 

~~ 

Bone marrow ......................... Pheochromocytoma malignant, 

metastatic, adrenal medulla X 

Total 

Tissues1 

Tumors 

50 

47 

47 

48 

49 

46 

43 

50 

2 

50 

50 

48 

49 

49 

50 

50 

1 

50 

1 

1 

50 

1 

46 

50 

10 

50 

1 

3 

50 

1 

50 

50 

1 

3 

50 

1 
128 Scopolamine Hydrobrimide Trihydrate, Nl" TR 445 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System (continued) 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Pheochromocytoma malignant, 

metastatic, adrenal medulla 

Thymus 

Integumentary System 

Mammary gland 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

MusculoskeletalSystem 

Bone 

Skeletal muscle 

Rhabdomyosarcoma 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Pheochromocytoma malignant, 

metastatic, adrenal medulla 

Nose 

Trachea 

Special Senses System 

None 

Urinary System 

Kidney 

Urinary bladder 

SystemicLesions 

Multiple organs 

Leukemia mononuclear 

0011223333333344445555566 

4579593334799978990271900 

1880412266212435892006037 

4444444444444444444444444 

I654254725674456582544233 

5145428390082353903647153 

......................... 

......................... 

.......................... 

......................... 

++++++++++++++++M++++++++ 

xx x 

......................... 

......................... + 

X 

......................... + 

+ 

.......................... 

......................... 

......................... 

......................... 

......................... 

......................... x x 
Lesions in Female Rats 129 

TABLEB2 

Individual Animal Tumor Pathology of Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System (continued) 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Pheochromocytoma malignant, 

metastatic, adrenal medulla 

Thymus 

Integumentary System 

Mammary gland 

Fibroadenoma 

Fibroadenoma, multiple 

Skin 

Musculoskeletal System 

Bone 

Skeletal muscle 

Rhabdomyosarcoma ~~ 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Pheochromocytoma malignant, 

metastatic, adrenal medulla 

Nose 

Trachea 

Special Senses System 

None 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Leukemia mononuclear 

6667777777777777777777777 

2462222222222222222222222 

9736666666688888888888888 

4444444444444444444444444 Total 

6332335667722333445566677 Tissues/ 

8085198452478467093726907 Tumors 

......................... 50 

......................... 50 

......................... 50 

X 1 

......................... 50 

......................... 49 

X x x x xxxx X 12 

X xx 3 

......................... 50 

......................... 50 

1 

1 

......................... 50 

+ 2 

+ 2 

......................... 50 

X 1 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

X X 4 
130 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEB3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate 

ClitoralGland:Adenoma 

Overall ratea 

Adjusted rateb 

Terminal rate' 

First incidence (days) 

Life table testd 

Logistic regression testd 

Cochran-Armita e testd 

Fisher exact test %

MammaryGland:Fibroadenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

' Logistic test regression 

Cochran-Armitage test 

Fisher exact test 

MammaryGland:Fibroadenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

VehicleControl 1 mgkg 

3/49 (6%) 2/60 (3%) 3/48 (6%) 1/50 (2%) 

9.1X 11.8% 10.4% 4.5% 

3/33 (9%) 2/17 (12%) 2/24 (8%) 1/22 (5%) 

P=0.337N 

726 (T) 

P=O.578 

726 (T) 

P=O.532 

547 

P=0.458N 

726 ("7 

P=0.350N P=O.578 P=O.620 P=0.458N 

P=0.301N 

P=0.404N P~O.651 P=0.301N 

29/50 (58 46) 10160 (17%) 19/50 (38%) 15/50 (30%) 

70.4% 46.1 % 59.9% 57.2% 

22/34 (65 %) 6/17 (35%) 14/26 (54%) 11/22 (50%) 

562 562 404 576 

P=0.387N P=0.134N P=0.242N P=0.249N 

P=0.396N P=0.029N P=0.061N P=0.149N 

P=0.224N 

P<0.001N P=0.036N P=0.004N 

or Adenoma 

29/50 (58%) 10160 (17%) 20/50 (40%) 15/50 (30%) 

70.4% 46.1%. 63.2% 57.2% 

22/34 (65 %) 6/17 (35%) 15/26 (58%) 11/22 (50%) 

562 562 404 576 

P=0.375N P=0.134N P=0.316N P=0.249N 

P=0.385N P=0.029N P=0.092N P=0.149N 

P=0.215N 

P<0.001N P=0.055N P=0.004N 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

ThyroidGland(C-cell):Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

PituitaryGland(Pars Distaliis): Adenoma 

20/50 (40%) 

50.6% 

15/34 (44%) 

46 1 

P=0.207N 

P=0.158N 

P=0.106N 

3/50 (6%) 

8.8% 

3/34 (9%) 

726 (T) 

P=0.448N 

P=0.448N 

P=0.409N 

13/60 (22%) 

51.0% 

6/17 (35%) 

408 

P-0.320 

P=0.493N 

P=0.030N 

1/60 (2%) 

5.9% 

1/17 (6%) 

726 (T) 

P=0.572N 

P=0.572N 

P=0.244N, 

14/50 (28%) 

38.1 % 

5/26 (19%) 

547 

P=0.382N 

P=0.174N 

P=0.146N 

2/50 (4%) 

7.7% 

2/26 (8%) 

726 (T) 

P=0.622N 

P=0.622N 

P=0.500N 

10150 (20%) 

38.6% 

7/22 (32%) 

520 

P=0.280N 

P=0.164N 

P=0.024N 

1/50 (2%) 

4.5 % 

1/22 (5%) 

726 (T) 

P=0.470N 

P=0.470N 

P=0.309N 

1 
- - 

Lesions in Female Rats 131 

TABLEB3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle Control 1 mgkg 25 mgkg 

ThyroidGland (Ccell): Adenoma or Carcinoma 

Overall rate 5/50 (10%) 2/60 (3 %) 2/50 (4%) 1/50 (2%) 

Adjusted rate 14.7% 11.8% 7.7% 4.5% 

Terminal rate 5/34 (15%) 2/17 (12%) 2/26 (8 %) 1/22 (5%) 

First incidence (days) 726 (T) 726 (T) 726 (T) 726 (T) 

Life table test P=0.214N P=0.557N P=0.334N P=0.226N 

Logistic regression test P=0.214N P=0.557N P=0.334N P=0.226N 

Cochran-Armitage test P=0.188N 

Fisher exact test P=0.151N P=0.218N P=0.102N 

ThyroidGland(FollicularCell):Adenoma 

Overall rate 1/50 (2%) 0160 (0%) 0150 (0%) 3/50 (6%) 

Adjusted rate 2.9% 0.0% 0.0% 13.6% 

Terminal rate 1/34 (3%) 0/17 (0%) 0126 (0%) 3/22 (14%) e First incidence (days) 726 (T) 726 (T) 

Life table test P=O.O28 P=0.638N P=0.554N P=O.164 

Logistic regression test P=O.O28 P=0.638N P=0.554N P=O.164 

Cochran-Armitage test P-0.037 

Fisher exact test P=0.455N P=0.500N P=O.309 

Uterus: Stromal Polyp 

Overall rate 6/50 (12 %) 1/60 (2%) 4/50 (8 X) 3/50 (6%) 

Adjusted rate 17.6% 2.6% 13.5% 10.8% 

Terminal rate 6/34 (18%) 0117 (0%) 3/26 (12%) 1/22 (5%) 

First incidence (days) 726 (T) 379 47 1 502 

Life table test P=0.582N P=0.218N P=0.529N P=0.484N 

Logistic regression test P=0.571N P=0.138N P=0.417N P=0.419N 

Cochran-Armitage test P=0.522N 

Fisher exact test P=0.033N P=0.370N P=0.243N 

All Organs: MononuclearCellLeukemia 

Overall rate 20/50 (40%) 6/60 (10%) 13/50 (26%) 4/50 (8 %) 

Adjusted rate 47.7% 26.9% 38.8% 13.9% 

Terminal rate 13/34 (38%) 2/17 (12%) 7/26 (27 %) 2/22 (9%) 

First incidence (days) 374 519 555 392 

Life table test P=0.022N P=0.138N P=0.307N P=O.OllN 

Logistic regression test P=0.013N P=0.013N P=0.114N P<0.001N 

Cochran-Armitage test P=0.009N 

Fisher exact test P<0.001N P=O.lOlN P<O.OOlN 

All Organs: BenignNeoplasms 

Overall rate 38/50 (76%) 19/60 (32%) 32/50 (64%) 29/50 (58%) 

Adjusted rate 84.3% 65.7% 83.5% 87.7% 

Terminal rate 27/34 (79%) 8/17 (47%) 20/26 (77 %) 18/22 (82%) 

First incidence (days) 461 379 404 291 

Life table test P=O.176 P=0.486N P=O.474 P=O.213 

Logistic regression test P=O.168 P=0.025N P=0.214N P=O.541 

Cochran-Amitage test P=O.404 

Fisher exact test P<0.001N P=0.138N P=0.044N 
132 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEB3 

Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control i mgkg 

All Organs:MalignantNeoplasms 

Overall rate 23/50 (46%) 9/60 (15%) 16/50 (32%) 6/50 (12%) 

Adjusted rate 55.1% 41.5% 45.7% 20.3% 

Terminal rate 16/34 (47%) 5/17 (29%) 8/26 (31 %) 3/22 (14%) 

First incidence (days) 374 519 555 391 

Life table test P=0.023N P=0.270N P=0.362N P-0.015N 

Logistic regression test P=0.012N P-0.052N P=0.133N P<O.OOiN 

Cochran-Annitage test P=0.008N 

Fisher exact test P<0.001N P=0.109N P<0.001N 

All Organs:Benign or MalignantNeoplasms 

Overall rate 48/50 (96%) 23/60 (38%) 39/50 (7846) 32/50 (64%) 

Adjusted rate 96.0% 78.4% 92.7% 88.6% 

Terminal rate 32/34 (94%) 11/17 (65%) 23/26 (88%) 18/22 (82%) 

First incidence (days) 374 379 404 291 

Life table test P=O.412 P=0.367N P=0.557N P=O.520 

Logistic regression test P=0.566N P<0.001N P=0.017N P=0.013N 

Cochran-Armitage test P=0.346N 

Fisher exact test P<0.001N P=0.007N P<0.001N 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animaldnumber of animals examined. Denominator is number of animals examined microscopically for clitoral gland, 

pituitary gland, thyroid gland, and uterus; for other tissues, denominator is number of animals necropsied. 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence at terminal kill 

Beneath the control incidence are the P values associated with the trend test. Beneath the dosed group incidence are the P values corresponding to 

painvise 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 neoplasmsinanimal group 
Lesions in Female Rats 133 

TABLE B4a 

Historical Incidence of Pituitary Gland (Pars Distalis) Adenoma 

in Female F344/N Rats Administered Water by Gavagea 

Incidence in Controls 

Overall Historical Incidence 

Total 170/365 (46.6%) 

Standard deviation 6.7% 

Range 42 96-5896 

a Data as of 17 June 1994 

TABLE B4b 

Historical Incidence of Mononuclear Cell Leukemia in Female M44/N Rats Administered Water by Gavagea 

Incidence in Controls 

Overall Historical Incidence 

Total 99/368 (26.9%) 

Standard deviation 7.6% 

Range 16%-40% 

a Data as of 17 June 1994; includes data for lymphocytic, monocytic, and undifferentiated leukemia 

TABLE B4c 

Historical Incidence of Thyroid Gland (Follicular Cell) Adenoma in Female M44/N Rats Administered Water by Gavagea ~ ~ ~ ~ 

Incidence in Controls 

Overall Historical Incidence 

Total 5/367 (1.4%) 

Standard deviation 1.9% 

Range 0r-4x 

a Data as of 17 June 1994 
134 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEB5 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydratea 

ControlVehicle 1 mgkg 5 mgkg 25 mgkg 

DispositionSummary 

Animals initially in study 

15-Month interim evaluation 

Early deaths 

Accidental deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

AlimentarySystem 

Intestine large, rectum 

Parasite metazoan 

Liver 

Basophilic focus 

Fatty change 

Hematopoietic cell proliferation 

Hepatodiaphragmatic nodule 

Inflammation, acute, focal 

Inflammation, granulomatous 

Pancreas 

Acinus, atrophy 

Cardiovascular System 

Heart 

Myocardium, degeneration 

Endocrine System 

Pituitary gland 

cyst 

Pars distalis, angiectasis 

Pars distalis, hyperplasia 

Thyroid gland 

C-cell, hyperplasia 

Follicular cell, hyperplasia 

GenitalSystem 

Ovary 

cyst 

RespiratorySystem 

Lung 

Alveolar epithelium, hyperplasia 

60 60 

10 

18 

13 10 

3 15 

34 17 

60 60 

1 (10%) 

3 (30%) 

1 (10%) 

1 (10%) 

(10) 

1 (10%) 

(10) 

60 

10 

2 

6 

16 

26 

60 

2 (20%) 

1 (10%) 

60 

10 

5 

7 

16 

22 

60 

4 (40%) 

a Number of animals examined microscopically at the site and the number of animals with lesion 
Lesions in Female Rats 135 

TABLEB5 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle Control 1 mgkg 5 mgkg 

15-Month Interim Evaluation(continued) 

Urinary System 

Kidney (10) 

Nephropathy 2 (20%) 

Urinary bladder (10) 

Calculus, gross observation 1 (10%) 

Transitional epithelium, hyperplasia 1 (10%) 

Systems Examined With No Lesions Observed 

General Body System 

HematopoieticSystem 

IntegumentarySystem 

MusculoskeletalSystem 

Nervous System 

Special Senses System 

2-Year Strcdy 

AlimentarySystem 

Intestine large, colon 

Parasite metazoan 

Intestine large, rectum 

Parasite metazoan 

Intestine small, duodenum 

Inflammation, chronic 

Inflammation, chronic 

Intestine small, jejunum (49) (47) 

Basophilic focus 

Liver (50) 

20 (40%) 

Clear cell focus 1 (2%) 

Eosinophilic focus 3 (6%) 

Fatty change 2(4%) 

Hepatodiaphragmatic nodule 11 (22%) 

Hyperplasia, adenomatous 1 (2%) 

Mixed cell focus 1(2%) 

Necrosis 1 (2%) 

Bile duct, inflammation, chronic 1 (2%) 

Serosa, fibrosis 

Fat, necrosis 

Mesentery (3) 

2 (67%) 

Lymphatic, cyst 1 (33%) 

Pancreas (50) 

Acinus, atrophy 8 (16%) 

Artery, inflammation, chronic 

Duct, concretion 

Salivary glands 

Inflammation, chronic 

Inflammation,subacute 

Hyperkeratosis 

Stomach,forestomach (60) 

Ulcer 1 (2%) 

25 mgkg 

(50) 

21 (42%) 

5 (10%) 

7 (14%) 

1(2%) 

(2)

2 (100%) 

1(2%) 

(50) 
136 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEB5 

Summary of the Incidence of Nonneoplastic Lesions in FemaleRats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Alimentary System (continued) 

2-Year Study (continued) 

Stomach, glandular 

Artery,mineralization 

Ulcer 

(50) 

1 (2%) 

3 (6%) 

(60) 

Cardiovascular System 

Blood vessel 

Inflammation, granulomatous 

Mineralization 

Heart 

Atrium, thrombosis 

Myocardium, degeneration 

Myocardium, mineralization 

Valve, thrombosis 

EndocrineSystem 

Adrenal cortex 

Degeneration 

Hyperplasia 

Adrenal medulla 

Hyperplasia 

Bilateral, hyperplasia 

Pituitary gland 

cyst 

(50) 

33 (66%) 

(60) 

28 (47%) 

Pars distalis, hyperplasia 

Pigmentation, hemosiderin 

16 (32%) 

1 (2%) 

11 (18%) 12 (24%) 

C-cell, hyperplasia 

Thyroid gland (50) 

5 (10%) 

(60) 

1 (2%) 

(50) 

2 (4%) 

General Body System 

None 

Genital System 

Clitoral gland 

Ectasia 

Hyperplasia 

Inflammation, suppurative 

Ovary 

cyst 

uterus 

Decidual reaction 

Dilatation 

Hyperplasia, cystic 1(2%) 

Inflammation, chronic 1 (2%) 

Inflammation,suppurative 

Vagina 

Infiltration cellular, polymorphonuclear 

Epithelium, vacuolization cytoplasmic 1 (100%) 
Lesions in Female Rats 137 

TABLEB5 

Summary of the Incidence of Nonneoplastic Lesionsin Female Rats in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate (continued) . 

&Year Study (continued) 

Bone mamw 

HematopoieticSystem 

Inflammation, granulomatous 

Myelofibrosis 

Myelostromal proliferation 

Lymph node 

Renal. pigmentation, hemosiderin 

Lymph node, mandibular 

Inflammation, chronic 

Lymph node, mesenteric 

Inflammation,subacute 

Spleen 

Fibrosis 

Hematopoietic cell proliferation 2 (4%) 

Necrosis 

Lymphoid follicle, atrophy 1 (2%) 2 (4%) 

Red pulp, atrophy 1 (2%) 

IntegumentarySystem 

Mammary gland 

Edema 

Inflammation 

Hyperplasia, cystic 

Skin 

Acanthosis 

Ulcer 

Hyperkeratosis 

MusculoskeletalSystem 

Bone 

Osteopetrosis 

None 

Nervous System 

Respiratory System 

Erythrophagocytosis 

Lung 

1 (2%) 

(50) 

Foreign body 1 (2%) 1 (2%) 1 (2%) 

Hemorrhage 1 (2%) 1 (2%) 

Infiltrationcellular,mononuclearcell 1 (2%) 1 (2%) 

chronic Inflammation, 1 (2%) 1 (2%) 2 (4%) 

Inflammation,subacute 

Inflammation, granulomatous 

1 (2%) 

1 (2%) 

1 (2%) 

Alveolar epithelium, hyperplasia 2 (4%) 2 (3%) 3 (6%) 1 (2%) 

Alveolus, edema 1 (2%) 

Bronchus, inflammation, acute 1 (2%) 
138 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEB5 

Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle Control 1 mgflrg 5 mgflrg 25 mgkg 

2-Year Study (continued) 

Respiratory System (continued) 

Nose (50) 

Exudate 

Inflammation,suppurative 

Special Senses System 

Ear (2) (1)

External ear, ulcer 1 (100%) 

EYe (2) (3) (2) 

Inflammation, chronic 1 (33%) 

Anterior chamber, inflammation 1 (50%) 

Cornea.necrosis 1 (50%) 

Urinary System 

Kidney (50) (60) 

cyst 1 (2%) 

Hemorrhage 1 (2%) 1 (2%) 

Hydronephrosis 1 (2%) 

Infarct 1 (2%) 

Nephropathy 33 (66%) 22 (37%) 27 (54%) 13 (26%) 

Pelvis, inflammation, chronic 1 (2%) 

Urinary bladder (50) (60) (49) (50) 

Transitional epithelium, hyperplasia 1 (2%) 
139 

APPENDIX C 

SUMMARY OF LESIONS IN MALE MICE 

IN THE 2-YEAR GAVAGE STUDY 

OF SCOPOLAMINE HYDROBROMIDE TRIHYDRATE 

TABLEC1 Summary of the Incidence of Neoplasms in Male Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate . . ... . . . .. . 141 

TABLEC2 Individual Animal Tumor Pathology of Male Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate . . . . . ... .. . 146 

TABLEC3 Statistical Analysis of Primary Neoplasms in Male Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate . . .. . . . . . .. 168 

TABLEC4 Historical Incidence of Hepatocellular Neoplasms 

in Male B6C3Fl Mice Administered Water by Gavage . . . .. . . . . . . . . . . . .. . . .. 173 

TABLEC5 Summary of the Incidence of Nonneoplastic Lesions in Male Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate . . . . . .... .. 174 
140 .Scopolamine Hydrobromide Trihydrate, NTP TR 445 
Lesions 141 in Male Mice 

of Scopolamine Hydrobromide Trihydratea 

Summary of the Incidence of Neoplasms in Male Mice in the 2-Year Gavage Study 

TABLEC1 

Early deaths 

15-Month interim evaluationb 

Animals initially in study 

Disposition Summary 

Natural deaths 

Moribund 

Accidental deaths 

Terminal sacrifice 

Survivors 

20 

70 

6 

4 

40 

20 

70 

4 

7 

39 

20 

70 

5 

4 

2 

39 

20 

70 

2 

7 

2 

39 

Animals examined microscopically 60 60 60 60 

15-Month Interim Evaluation 

Liver 

Alimentary System 

Hepatocyte, hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Hepatocellular adenoma 

Hepatocellular carcinoma 

(10) 

1 (10%) 

1 (10%) 

(10) 

1 (10%) 

1 (10%) 

2 (20%) 

(10) 

1 (10%) 

(10) 

1 (10%) 

Lung 

Respiratory System 

AlveoIar/bronchiolar adenoma 

(10) 

2 (20%) 

(10) 

Urinary System 

Special Senses System 

Nervous System 

Musculoskeletal System 

Integumentary System 

Hematopoietic System 

Genital System 

General Body System 

Endocrine System 

Cardiovascular System 

Systems Examined WithNo Neoplasm Observed 
142 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC1 

Summary of the Incidence of Neoplasms in Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 

2-Year Study 

AlimentarySystem 

Intestine small, duodenum 

Adenoma 

Intestine small, jejunum 

Carcinoma 

Intestine small, ileum 

Liver 

Carcinoma, multiple 

Hemangioma 1 (2%) 

Hemangiosarcoma 

Hemangiosarcoma, multiple 

Hemangiosarcoma, metastatic, skin 1 (2%) 

Hemangiosarcoma, metastatic, spleen 

Hepatoblastoma 

Hepatocellular carcinoma 6 (12%) 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 10 (20%) 

Hepatocellular adenoma, multiple 16 (32%) 

Histiocytic sarcoma 3(6%) 

It0 cell tumor NOS, multiple 1(2%) 

Mesentery (4)

Hemangiosarcoma 

Histiocytic sarcoma 

Pancreas 

Stomach, forestomach 

Mast cell tumor malignant 

Squamous cell papilloma 

Tooth (14)

Odontoma 1 (7%) 

Cardiovascular System 

Heart (50)

Hemangiosarcoma, metastatic, spleen 

EndocrineSystem .. 

1 mgk 5 mgkg 

1 (2%) 

1(2%) 

13 (26%) 5 (lo%), 

2(4%) 

18 (36%) 6 (12%) 

4 (8%) 3 (6%) 

(3) 

1(33%) 

1(2%) 

(4) (4) (2) 

1 (25%) 2 (50%) 1 (50%) 

(50) 

25 mgkg 

7 (14%) 

8 (16%) 

1 (2%) 

Islets, pancreatic (50) (50) (50) (50)

Adenoma 3(6%) 1(2%) 1(2%) 

Pituitary gland 

Follicular cell, adenoma 

Pars intermedia, adenoma 

Pars distalis, adenoma 

Thyroid gland 

(48) 

1 (2%) 

(50) 

(47) 

(50) 

1 (2%) 

(46) 

1 (2%) 

(50) 

(44) 

1 (2%) 

(50) ’ 

1(2%) 

None 

General Body System 
Lesions in Male Mice 143 

TABLEC1 

Summary of the Incidence of Neoplasms in Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

&Year Study (continued) 

Genital System 

Preputial gland 

Sarcoma 

Seminal vesicle 

Adenoma 

Testes 

Interstitial cell, adenoma 

Hematopoietic System 

Bone marrow 

Hemangiosarcoma, metastatic, liver 

Hemangiosarcoma,metastatic,skin 

Lymph node 

Bronchial, carcinoma, metastatic, liver 

Mediastinal, carcinoma, metastatic, liver 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Carcinoma, metastatic, liver 

Hemangiosarcoma, metastatic, spleen 

Histiocytic sarcoma 

Spleen 

Hemangiosarcoma 

Hemangiosarcoma, multiple 

Hemangiosarcoma,metastatic,liver 

Hemangiosarcoma,metastatic,skin 

Histiocytic sarcoma 

Thymus (40) 

Carcinoma, metastatic, liver 1 (3%) 

Integumentary System 

Skin (50) (50) (50) (50) 

Mast cell tumor malignant 1 (2%) 

Subcutaneous tissue, hemangiosarcoma 1 (2%) 

Musculoskeletal System 

None 

Nervous System 

None 
144 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC1 

Summary of the Incidence of Neoplasms in Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle Control 1 mgkg 5 mgkg 25 mgkg 

2-Year Study (continued) 

RespiratorySystem 

Lung

Alveolarlbronchiolar adenoma 

Alveolar/bronchiolar adenoma, multiple 

Alveolar/bronchiolar carcinoma 

Alveolarhronchiolar carcinoma, multiple 

Carcinoma, metastatic, harderian gland 2 (4%) 

Carcinoma. metastatic, liver 1 (2%) 

Hepatocellular carcinoma, metastatic, liver 3 (6%) 1 (2%) 3 (6%) 

Histiocytic sarcoma 1 (2%) 

Nose (50)

Mast cell tumor malignant 1 (2%) 

Special Senses System 

Harderian gland 

Adenoma 

Carcinoma 

Urinary System 

Kidney (50) (50)

Histiocytic sarcoma 1 (2%) 

Artery, hepatocellular carcinoma, metastatic. 

liver 

Renal tubule, adenoma 

Renal tubule, carcinoma 2 (4%) 

Urinary bladder (50) (50) 

Systemic Lesions 

Multiple organs' (50) (50) (50) (50)

Histiocytic sarcoma 3 (6%) 1 (2%) 

Leukemia lymphocytic 1 (2%) 1 (2%) 1 (2%) 

Lymphornil malignant lymphocytic 1 (2%) 1 (2%) 3 (6%) 

Lymphoma malignant mixed 3 (6%) 2 (4%) 1 (2%) 
Lesions in Male Mice 

TABLEC1 

Summary of the Incidence of Neoplasms in Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 1 mg/kg 5 25 mgkg 

Neoplasm Summary 

~0ta1animals with primary neoplasmsd 

l5-Month interim evaluation 4 4 1 2 

2-Year Study 43 41 31 29 

Total primary neoplasms 

15-Month interim evaluation 4 4 1 2 

2-Year study 69 66 43 37 

Total animals with benign neoplasms 

15-Month interim evaluation 4 4 1 1 

2-Year study 36 28 21 18 

Total benign neoplasms 

15-Month interim evaluation 4 4 1 1 

2-Year study 48 34 26 20 

Total animals with malignant neoplasms 

15-Month interim evaluation 1 

2-Year study 17 25 16 15 

Total malignant neoplasms 

l5;Month interim evaluation 1 

2-Year study 20 32 17 17 

Total animals with metastatic neoplasms 

2-Year study 4 5 4 3 

Total metastatic neoplasms 

2-Year study 6 6 10 4 

Total animals with uncertain neoplasmsbenign or malignant 

2-Year study 1 

Total uncertain neoplasms 

2-Year study 1 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

Ten animals per group received ophthalmic examinations and were discirded without further evaluation 

Number of animals with any tissue examined microscopically 

Primary neoplasms: all neoplasms except metastatic neoplasms 
146 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC2 

Individual Animal Tumor Pathologyof Male Mice in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: Vehicle Control 

Number of Dayson Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Adenoma 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hemangioma 

Hemangiosarcoma, metastatic, skin 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Histiocytic sarcoma 

Ito cell tumor NOS, multiple 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tooth 

Odontoma 

Cardiovascular System 

Blood vessel 

Heart 

EndocrineSystem 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars intemedia, adenoma 

Thyroid gland 

General Body System 

None 

+: Tissue examined microscopically 

A: Autolysis precludes examination 

2456666177177717771177711 

9444889002222222222222222 

4101034075666666666777711 

0000000000000000000000000 

1610351532002455666011222 

6942969452560208145901356 

......................... 

......................... 

+M+++++++++++++++++++++++ 

......................... 

......................... 

......................... 

.......................... 

......................... 

......................... X 

X 

xx x X X 

X X X X 

x x xx X xxx 

X 

+ ......................... 

......................... 

......................... 

......................... + + + + 

X 

......................... 

......................... 

......................... 

......................... 

......................... X 

++++++++++++M++++++++++++ 

++++M++++++++++M+++++++++ 

X 

......................... 

M:Missingtissue X: Lesion present 

I: Insufficient tissue Blank: Not examined 
Lesions in Male Mice I47 

TABLEC2 

Individual Animal Tumor Pathologyof Male Mice in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: Vehicle Control (continued) 

Number of Dayson Study 

0000000000000000000000000 Total 

Carcass ID Number 3334444455560012233344561 Tissues/ 

2310518951963151906816230 Tumors 

AlimentarySystem 

Esophagus ......................... 50 

Gallbladder +++++++++++++M+++++++++++ 49 

Intestine large, colon ......................... 49 

Intestine large, rectum ........................... 50 

Intestine large, cecum +++++++++++++++++++M+++++ 49 

Intestine small, duodenum ......................... 50 

Adenoma X 1 

Intestine small, jejunum ......................... 50 

Intestine small, ileum +++++++++++++++++++++++M+ 49 

Liver ......................... 50 

Hemangioma 1 

Hemangiosarcoma, metastatic, skin 1 

Hepatocellular carcinoma X 6 

Hepatocellular adenoma X xxxx X 10 

Hepatocellular adenoma, multiple x x X x x X 16 

Histiocytic sarcoma X X 3 

It0 cell tumor NOS, multiple X 1 

Mesentery + + + 4 

Pancreas ......................... 50 

Salivary glands ......................... 50 

Stomach, forestomach ......................... 50 

Stomach, glandular ......................... 50 

Tooth + + ++++ + ++ + 14 

Odontoma 1 

Cardiovascular System 

Blood vessel ......................... 50 

Heart ......................... 50 

EndocrineSystem 

Adrenal cortex ......................... 50 

Adrenal medulla ++++++++++++++++++++++M++ 49 

Islets, pancreatic ......................... 50 

Adenoma x x 3 

Parathyroid gland ++++M+++++++++++++++M++M+ 46 

Pituitary gland ......................... 48 

Pars intermedia, adenoma 1 

Thyroid gland ......................... 50 

General Body System 

None 
Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study 

of ScopolamineHydrobromide Trihydrate: Vehicle Control (continued) 

2456666117711711111717711 

Number of Days on Study 944488900222222222222.2222 

4101034015666666666711711 

0000000000000000000000000 

Carcass ID Number 1610351532002455666011222 

6942969452560208145901356 

Genital System 

Epididymis ......................... Penis + + 

Preputial gland ......................... Prostate ......................... Seminal vesicle ......................... Testes ......................... 

HematopoieticSystem 

Bone marrow ......................... Hemangiosarcoma, metastatic, skin X 

Lymph node + 

Lymph node, mandibular ......................... Lymph node, mesenteric +++++++++++++M+++++++++++ 

Histiocytic sarcoma X 

Spleen ......................... Hemangiosarcoma, metastatic, skin X 

Histiocytic sarcoma A 

Thymus +++M+++++M+++M+++++++++M+ 

IntegumentarySystem 

Mammary gland MMMMMMMMMMMMM+MMMMMM+MMMM 

Skin ......................... Subcutaneous tissue, hemangiosarcoma X 

Musculoskeletal System 

Bone ......................... 

Nervous System 

Brain ......................... 

Respiratory. System 

Lung ......................... Alveolarlbronchiolar adenoma xx x x x X 

Alveolar/bronchiolar adenoma, 

Alveolar/bronchiolar carcinoma 

multiple 

X X 

Alveolar/bronchiolar carcinoma, 

multiple X 

Hepatocellular carcinoma, metastatic, 

liver xx 

Histiocytic sarcoma X 

Nose ......................... Trachea . ......................... 

Special Senses System 

Harderian gland MM++M++M+MMMMMM++++MMMMMM 

Adenoma X 
Lesions in Male Mice 149 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: Vehicle Control (continued) 

Number of Days on Study 

Carcass ID Number 

Genital System 

Epididymis 

Penis 

Preputial gland 

Prostate 

Seminal vesicle 

Testes 

Hematopoietic System 

Bone marrow 

Hemangiosarcoma, metastatic, skin 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Hemangiosarcoma, metastatic, skin 

Histiocytic sarcoma 

Thymus 

Integumentary System 

Mammary gland 

Skin 

Subcutaneous tissue, hemangiosarcoma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar adenoma, 

multiple 

Alveolar/bronchiolar carcinoma 

Alveolar/bronchiolar carcinoma, 

multiple 

Hepatocellular carcinoma, metastatic, 

liver 

Histiocytic sarcoma 

Nose 

Trachea 

Special Senses System 

Harderian gland 

Adenoma 

7777777777777777777771777 

2222222222222222222222222 

I777777777779999999999999 

0000000000000000000000000 Total 

3334444455560012233344567 Tissues/ 

2370578957963757906816230 Tumors 

......................... 50 

2 

......................... 50 

++++++++++++++++++M++++++ 49 

......................... 50 

......................... 50 

......................... 50 

1 

+ 2 

......................... 49 

......................... 48 

X X 3 

......................... 50 

1 

X 2 

+M+++M+M++++++++++++M++++ 42 

MMMMMMMMMMMMMMMMMMMMMMMMM 2 

......................... 50 

1 

......................... 50 

......................... 50 

......................... 50 

x xx X X 11 

X 1 

2 

1 

X 3 

1 

......................... 50 

......................... 50 

+M+MMMMM+MM+MMM+MMM+MMMM+ 16 

X X 3 
150 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE C2 

Individual Animal Tumor Pathologyof Male Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

2456666777777177777777777 

Number of Days on Study 9444889002222222222222222 

4707034075666666666177777 

000000000000000~0000000000 

Carcass ID Number 1610351532002455666011222 

6942969452560208145901356 

Urinary System 

Kidney .......................... Histiocytic sarcoma 

Renal tubule, carcinoma X X 

Urinary bladder ......................... 

Systemic Lesions 

Multiple organs ......................... Histiocytic sarcoma X 

Leukemia lymphocytic X 

Lymphoma malignant lymphocytic X 

Lymphoma malignant mixed X 
Lesions in Male Mice 151 

TABLEC2 

Individual Animal Tumor Pathologyof Male Mice in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

7777777777777777777777777 

Number of Days on Study 2222222222222222222222222 

7777777777779999999999999 

0000000000000000000000000 

Carcass ID Number 3334444455560012233344567 

2370578957963757906816230 

Urinary System 

Kidney ......................... Histiocytic sarcoma X 

Renal tubule, carcinoma 

Urinary bladder ......................... 

Systemic Lesions 

Multiple organs ......................... Histiocytic sarcoma X X 

Leukemia lymphocytic 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed X X 

Total 

Tissues1 

Tumors 

50 

1 

2 

50 

50 

3 

1 

1 

3 
152 Scopolamine Hydrobromiae Trihydrate, NTP TR 445 

TABLE C2 

Individual Animal Tumor Pathologyof Male Mice in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate: 1 mg/kg 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hemangiosarcoma, metastatic, spleen 

Hepatoblastoma 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Mesentery 

Hemangiosarcoma 

Pancreas 

Salivary glands 

Stomach, forestomach 

Mast cell tumor malignant 

Stomach, glandular 

Tooth 

Odontoma 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Epididymis 

Preputial gland 

Prostate 

Seminal vesicle 

Adenoma 

Testes 

Interstitial cell, adenoma 

4566666111111111111111711 

0401669000022222222222222 

5148392511966666661711111 

1111111111111111111111111 

3364155411636189993445555 

1665052047241312562491418 

~~ ~ ~~ ~ 

......................... 

......................... 

......................... 

......................... 

++M++++++++++++++++++++++ 

......................... 

......................... 

......................... 

......................... X 

xxx X xxx x x X 

x x 

xx xx x xx X 

+ 

X 

......................... 

......................... 

......................... 

......................... + + 

......................... 

......................... 

......................... 

+++++++++++M+++++M+++++++ 

......................... 

+++++++M+++++MM++M+++++++ 

++++M++++++M+++++++M+++++ 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 
Lesions in Male Mice 153 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 1mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hemangiosarcoma, metastatic, spleen 

Hepatoblastoma 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Mesentery 

Hemangiosarcoma 

Pancreas 

SalivaIy glands 

Stomach, forestomach 

Mast cell tumor malignant 

Stomach, glandular 

Tooth 

Odontoma 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Epididymis 

Preputial gland 

Prostate 

Seminal vesicle 

Adenoma 

Testes 

Interstitial cell, adenoma 

7777777777777777777777~77 

2222222222222222222222222 

7777777777799999999999999 

1111111111111111111111112 Total 

6617788889933346677788890 Tissues/ 

8928934793913583715602840 Tumors 

......................... 50 

+++++++++++++M+++++++++++ 49 

......................... 50 

......................... 50 

......................... 49 

......................... 50 

......................... 50 

++++++M++++++++++++++++++ 49 

......................... 50 

1 

X 1 

X X X 13 

2 

x xx xxxxx xx 18 

X xx x 4 

+ + 3 

1 

......................... 50 

......................... 50 

......................... 50 

1 

......................... 50 

+ + 4 

X 1 

......................... 50 

......................... 50 

......................... 50 

+M+M++++++M++++++++++++++ 45 

......................... 50 

++MM++++++++M++++++M+++M+ 41 

......................... 47 

......................... 50 

X 1 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

X 1 

......................... 50 

X 1 
154 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 1mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Bone marrow 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma, multiple 

Thymus 

Integumentary System 

Mammary gland. 

Skin 

Mast cell tumor malignant 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolarlbronchiolar carcinoma 

Carcinoma, metastatic, harderian 

gland 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Mast cell tumor malignant 

Trachea 

Special Senses System 

EY e Harderian gland 

Adenoma 

Carcinoma 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Lymphoma malignant mixed 

4566666111111111111111111 

0401669000022222222222222 

5148392511966666661711117 

1111111111111111111111111 

3364155411636189993445555 

1665052047241312562491418 

......................... 

+++++++++++++++++++++++M+ 

++++++++++++++++M++++++++ 

......................... X 

M++MMM+++M+++++++++++++++ 

MMMMMMMMMMMMMMMMMMMMMMMMM 

......................... X 

......................... 

......................... 

......................... x xx x 

X 

X X 

xx X 

......................... X 

......................... 

+ + 

MMM+MM++++++MMM+MM+M+++M+ 

X X X X 

......................... 

......................... 

......................... 
Lesions in Male Mice 155 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 1mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

HematopoieticSystem 

Bone marrow 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma, multiple 

Thymus 

IntegumentarySystem 

Mammary gland 

Skin 

Mast cell tumor malignant 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, harderian 

gland 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Mast cell tumor malignant 

Trachea 

Special Senses System 

EY e Harderian gland 

Adenoma 

Carcinoma 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Lymphoma malignant mixed 

7777777777777777777777777 

2222222222222222222222222 

7777777777799999999999999 

1111111111111111111111112 Total 

6671788889933346677788890 Tissues/ 

8928934793913583715602840 Tumors 

......................... 50 

+++++++++++++++++++++M+++ 48 

+++M+++++++M+++++++M+++++ 46 

......................... 50 

1 

+++++++++t+++++++M+++++++ 44 

MMMMMMMMMMMMMMMMMMMMMMMMM 

......................... 50 

1 

......................... 50 

......................... 50 

......................... 50 

x x X 7 

X x x 4 

2 

3 

......................... 50 

X 2 

......................... 50 

+ 3 

MMMMM+M+M+MM+MMMMMMMMM+MM 18 

X 1 

4 

......................... 50 

......................... 50 

......................... 50 

X X 2 
156 Scopolamine Hydrobromide Trihydrate, NTP TR 445

TABLE C2

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg

o 1 3 5 5 5 5 6 6 6 7 7 7 7 7 7 7 7 7 7 7 7 777

Number of Days on Study 5 372 6 9 909 902 2 2 2 2 2 2 2 2 2 2 222

o 1 600 0 4 6 1 3 7 6 6 6 6 6 6 6 6 7 7 7 7 7 7

2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2

Carcass ID Number 105 1 3 742 5 5 3 002 4 4 4 5 600 1 122

9 4 4 7 2 0 2 8 2 1 967 1 6 7 8 323 846 5 9

Alimentary System

Esophagus + + + + + + + + + + + + + + + + + + + + + + + + +

Gallbladder M + + + + + + + + + + + + + + + + + + + + + + + +

Intestine large, colon + + + + + + + + + + + + + + + + + + + + + + + + +

Intestine large, rectum + + + + + + + + + + + + + + + + + + + + + + + + +

Intestine large, cecum + + + + + + + + + + + + + + + + + + + + + + + + +

Intestine small, duodenum + + + + + + + + + + + + + + + + + + + + + + + + +

Intestine small, jejunum + + + + M + + + + + + + + + + + + + + + + + + + +

Carcinoma X

Intestine small, ileum + + + + + + + + + + + + + + + + + + + + + + + + .+

Liver + + + + + + + + + + + + + + + + + + + + + + + + +

Carcinoma, multiple X

Hemangiosarcoma X X X

Hemangiosarcoma, multiple X

Hepatocellular carcinoma X X

Hepatocellular adenoma X X

Hepatocellular adenoma, multiple X X

Mesentery + +

Pancreas + + + + + + + + + + + + + + + + + + + + + + + + +

Salivary glands + + + + + + + + + + + + + + + + + + + + + + + + +

Stomach, forestomach + + + + + + + + + + + + + + + + + + + + + + + + +

Squamous cell papilloma X

Stomach, glandular + + + + + + + + + + + + + + + + + + + + + + + + +

Tooth + +

Odontoma X

Cardiovascular-System

Blood vessel + + + + + + + + + + + + + + + + + + + + + + + + +

Heart + + + + + + + + + + + + + + + + + + + + + + + + +

:Hemangiosarcoma, metastatic, spleen X

Endocrine System

Adrenal cortex + + + + + + + + + + + + + + + + + + + + + + + + +

Adrenal medulla + + + + + + + + + + + + + + + + + + + + + + + + +

Islets, panc'reatic + + + + + + + + + + + + + + + + + + + + + + + + +

Adenoma X

Parathyroid gland + + + + + + + + + + + + + + + M M + + + + + + + +

Pituitary gland + M + + + + + + + + + + + + + + + + + + + M + + +

Pars intermedia, adenoma X

Thyroid gland + + + + + + + + + + + + + + + + + + + + + + + + +

General Body System

None

Genital System

Epididymis + + + + + + + + + + + + + + + + + + + + + + + + +

Preputial gland + + + + + + + + + + + + + + + + + + + + + + + + +

Prostate + + + + + + + + + + + + + + + + + + + + + + + + +

Seminal vesicle + + + + + + + + + + + + + + + + + + + + + + + + +

Testes + + + + + + + + + + + + + + + + + + + + + + + + +

Interstitial cell, adenoma X
Lesions in Male Mice 157 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small. jejunum 

Carcinoma 

Intestine small, ileum 

Liver 

Carcinoma, multiple 

Hemangiosarcoma 

Hemangiosarcoma, multiple 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Squamous cell papilloma 

Stomach, glandular 

Tooth 

Odontoma 

Cardiovascular System 

Blood vessel 

Heart 

Hemangiosarcoma, metastatic, spleen 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars intermedia, adenoma 

Thyroid gland 

General Body System 

None 

Genital System 

Epididymis 

Preputial gland 

Prostate 

Seminal vesicle 

Testes 

Interstitial cell, adenoma 

7777777777777777777777777 

2222222222222222222222222 

7777777799999999999999999 

2222222222222222222222222 Total 

3344446600011223345556666 Tissues1 

5803496912508273615893457 Tumors 

......................... 50 

......................... 49 

......................... 50 

......................... 50 

+++++++++++M+++++++++++++ 49 

......................... 50 

......................... 49 

1 

......................... 50 

......................... 50 

1 

3 

1 

X X X 5 

X X X X 6 

X 3 

+ 3 

......................... 50 

......................... 50 

......................... 50 

1 

......................... 50 

+ + 4 

X 2 

......................... 50 

......................... 50 

1 

......................... 50 

......................... 50 

......................... 50 

1 

++++++++M++++++++++++++++ 47 

++++M+++++M++++++++++++++ 46 

1 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

1 
158 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

0135555666111111111111111 

Number of Days on Study 5372699099022222222222222 

0160004613766666666711111 

~~ ~~ ~~ ~ ~ ~~ 

2222222222222222222222222 

Carcass ID Number 1051314255300244456001122 

9441202821961161832384659 

HematopoieticSystem 

Bone marrow ......................... Hemangiosarcoma, metastatic, liver X 

Lymph node + + 

Bronchial, carcinoma, metastatic, 

liver X 

Mediastinal, carcinoma, metastatic, 

liver X 

Lymph node, mandibular +++M++M++++++++++++++++++ 

Lymph node, mesenteric +++++++M+++++M+++++++++++ 

Carcinoma, metastatic, liver X 

Hemangiosarcoma, metastatic, spleen X 

Spleen ......................... Hemangiosarcoma X 

Hemangiosarcoma, metastatic, liver X 

Thymus +++MMM+MM+M++++++++++++++ 

Carcinoma, metastatic, liver X 

IntegumentarySystem 

Mammary gland MMMMMMMMMMMMMMMMMMMMMMMMM 

Skin .......................... 

Musculoskeletal System 

Bone ......................... 

Nervous System 

Brain ......................... Peripheral nerve + 

Spinal cord + 

RespiratorySystem 

~~ ~ 

Lung ......................... Alveolarlbronchiolar adenoma x x X X 

Alveolar/bronchiolar adenoma, 

multiple X 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, liver X 

Hepatocellular carcinoma, metastatic, 

liver X 

Nose ......................... Trachea ......................... 

Special Senses System 

EYe 

Harderian gland +++MMM+MM+MM+MMMM+MM+MMM+ 

Adenoma X 

Carcinoma 
Lesions 159 in Male Mice 

TABLEC2 

Individual Animal Tumor Pathologyof Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Bone marrow 

Hemangiosarcoma, metastatic, liver 

Lymph node 

Bronchial, carcinoma, metastatic, 

liver 

Mediastinal, carcinoma, metastatic, 

liver 

Lymph node, mandibular 

Lymph node, mesenteric 

Carcinoma, metastatic, liver 

Hemangiosarcoma, metastatic, spleen 

Spleen 

Hemangiosarcoma 

Hemangiosarcoma, metastatic, liver 

Thymus 

Carcinoma, metastatic, liver 

Integumentary System 

Mammary gland 

Skin 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveoladbronchiolar adenoma, 

multiple 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, liver 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Trachea 

Special Senses System 

EY e Harderian gland 

Adenoma 

Carcinoma 

7777777777777777777777777 

2222222222222222222222222 

7777777799999999999999999 

2222222222222222222222222 Total 

3344446600011223345556666 Tissues/ 

5803496912508273615893457 Tumors 

......................... 50 

1 

2 

1 

1 

M++++++++++++++++++++++++ 47 

+++++++M+++++++++++++++++ 47 

1 

1 

......................... 50 

1 

1 

++++M+++++M+++++MM+++++++ 40 

1 

MMMMMMMMMMMMMMMMMMMMMMMMM 

......................... 50 

......................... 50 

......................... 50 

1 

1 

......................... 50 

xx X 7 

X 2 

X 1 

1 

1 

......................... 50 

......................... 50 

+ 1 

MMM++MM+M+M+M+MM+MM+MMM++ 19 

1 

X 1 
160 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC2 

Individual AnimalTumor Pathology of Male Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate: 5 mg/kg (continued) 

i 0135555666777777777717777 

Number of Days on Study 5372699099022222222222222 

0160004613766666666777777 

2222222222222222222222222 

Carcass ID Number 1051374255300244456001122 

9447202821967167832384659 

Urinary System 

Kidney 

Renaltubule,adenoma 

Urinarybladder 

Systemic Lesions 

Multiple organs 

Lymphoma malignant lymphocytic 

Leukemia lymphocytic 

......................... X 

X 

Lymphomamalignantmixed . 
Lesions in Male Mice 161 

TABLE C2 

Individual Animal Tumor Pathologyof Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

7777777777777777777777777 

Number of Days on Study 2222222222222222222222222 

7777777799999999999999999 

2222222222222222222222222 Total 

Carcass ID Number 3344446600011223345556666 Tissues1 

5803496912508273615893457 Tumors 

Urinary System 

Kidney ......................... 50 

Renal tubule, adenoma X 1 

Urinary bladder ......................... 50 

Systemic Lesions 

Multiple organs ......................... 50 

Leukemia lymphocytic 1 

Lymphoma malignant lymphocytic 1 

Lymphoma malignant mixed X 1 
162 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC2 

Individual Animal TumorPathology of Male Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate: 25 mg/kg 

Number of Dayson Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Histiocytic sarcoma 

Mesentery 

Histiocytic sarcoma 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tooth 

Odontoma 

Cardiovascular System 

Blood vessel 

Heart 

EndocrineSystem 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Follicular cell, adenoma 

~~ ~~ 

General Body System 

None 

GenitalSystem 

Epididymis 

Preputial gland 

Sarcoma 

Prostate 

Seminal vesicle 

Testes 

34445566611111111117171~1 

0003892290022222222222222 

0552742115766666666661111 

3232333323322223333332222 

2118201091211181223331899 

2413930611126166013519902 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

+++++++++++++++++M++++M++ 

......................... X X X 

xx X X 

X 

+ + 

X 

......................... 

......................... 

......................... 

......................... + + 

X 

......................... 

.......................... 

......................... 

......................... 

......................... 

+M+M+++M+++++++++++++++++ 

++++++++++++++++++M+++MM+ 

X 

......................... 

......................... 

......................... X 

......................... 

.......................... 

.......................... 
Lesions in Male Mice 163 

TABLEC2 

Individual Animal Tumor Pathologyof Male Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Histiocytic sarcoma 

Mesentery 

Histiocytic sarcoma 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Tooth 

Odontoma 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Epididymis 

Preputial gland 

Sarcoma 

Prostate 

Seminal vesicle 

Testes 

7777777777777777777777777 

2222222222222222222222222 

7777777777799999999999999 

2233333333322222233333333 Total 

9900012333377899901112234 Tissues/ 

3905884012815156792393540 Tumors 

......................... 50 

++++++++M+++++,+++++++++++ 49 

++++++++++++++++++++++M++ 48 

......................... 50 

......................... 49 

......................... 50 

......................... 50 

......................... 48 

......................... 50 

x x X X 7 

X x xx 8 

1 

2 

1 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

2 

1 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

X 1 

++++M+++++MM++++++++++M++ 43 

+++M++++++++M+++M++++++++ 44 

1 

......................... 50 

X 1 

......................... 50 

......................... 50 

1 

........................... 50 

......................... 50 

......................... 50 
164 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC2 

Individual AnimalTumor Pathology of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Carcass ID Number 

HematopoieticSystem 

Bone marrow 

Lymph node 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Histiocytic sarcoma 

Thymus 

IntegumentarySystem 

Mammary gland 

Skin 

Musculoskeletal System 

Bone 

NervousSystem 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Alveolarlbronchiolar adenoma 

Alveolar/bronchiolar adenoma, 

multiple 

Alveolarkxonchiolar carcinoma 

Hepatocellular carcinoma, metastatic, 

liver 

Histiocytic sarcoma 

Nose 

Mast cell tumor malignant 

Trachea 

Special Senses System 

Ear 

EY e Harderian gland 

Adenoma 

Carcinoma 

3444556667777717777771771 

000389229’0022222222222222 

0552742775766666666667777 

3232333323322223333332222 

2718201091277781223337899 

2413930617126766073579902 

......................... + + + 

X 

++++++++M++++++++++M+++++ 

........................ X ......................... X 

+++MM+++MM++++M+++M++++++ 

+MMMMMMMMMMMMMMMMMMMMMMMM 

......................... 

......................... 

......................... + 

+ 

......................... X 

X 

X 

......................... X 

......................... 

+ 

+ 

MM+MM+MMMMM+MM+M+MMMMMM+M 

X 

X 
Lesions in Male Mice 165 

TABLEC2 

Individual Animal Tumor Pathology of Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Bone marrow 

Lymph node 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Histiocytic sarcoma 

Spleen 

Histiocytic sarcoma 

Thymus 

Integumentary System 

Mammary gland 

Skin 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Alveoladbronchiolar adenoma 

Alveolar/bronchiolar adenoma, 

multiple 

Alveolar/bronchiolar carcinoma 

Hepatocellular carcinoma, metastatic, 

liver 

Histiocytic sarcoma 

Nose 

Mast cell tumor malignant 

Trachea 

Special Senses System 

Ear 

EY e Harderian gland 

Adenoma 

Carcinoma 

1117171177711111111117111 

2222222222222222222222222 

1717111117199999999999999 

2233333333322222233333333 Total 

9900012333371899901112234 Tissues/ 

3905884012815156192393540 Tumors 

......................... 50 

3 

1 

++++++++M++++++++++++M+++ 46 

......................... 41 

1 

......................... 50 

1 

++++M+++MM++++++++M++++++ 40 

MMMMMMMMMMMMMMMMMMMMMMMMM. 1 

......................... 50 

......................... 50 

......................... 50 

1 

1 

......................... 50 

X x x X 5 

X 1 

x x 2 

X X 3 

1 

......................... 50 

1 

......................... 50 

1 

+ 2 

MM++MMMM+MMMM+M+M+MM+MMMM 13 

X 2 

1 
166 Scopolamine Hydrobromide Trihydrate, N" TR 445 

TABLEC2 

Individual Animal Tumor Pathologyof Male Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate: 25 mg/kg (continued) 

3444556667777777777777777 

Number of Days on Study 0003892290022222222222222 

0552742775766666666667777 

3232333323322223333332222 

Carcass ID Number 2718201091277781223337899 

2413930617126766073579902 

~ ~~ ~~~ ~ 

Urinary System 

Kidney 

Artery, hepatocellular carcinoma, 

metastatic, liver 

Urinary bladder M++++++++++++++++++++++++ 

~ ~ ~~ ~ 

Multiple organs 

Systemic Lesions 

Lymphoma malignant 

Leukemia lymphocytic 

Histiocytic sarcoma 

lymphocytic 

.......................... X 

X 

X 
Lesions in Male Mice 167 

TABLEC2 

Individual Animal Tumor Pathologyof Male Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate: 25 mg/kg (continued) 

7777777777777777777777777 

Number of Days on Study 2222222222222222222222222 

7777777777799999999999999 

2233333333322222233333333 

Carcass ID Number 9900012333377899901112234 

3905884012815156792393540 

Urinary System 

Kidney ......................... Artery, hepatocellular carcinoma, 

metastatic, liver X 

Urinary bladder ......................... 

SystemicLesions 

Multiple organs ......................... Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Leukemia lymphocytic 

X X 

Total 

Tissues/ 

Tumors 

50 

1 

49 

50 

1 

1 

3 
168 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC3 

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate 

HarderianGland:Adenoma 

Overall ratea 

Adjusted rateb 

Terminal rate' 

First incidence (days) 

Life table testd 

Logistic regression testd 

Cochran-Armita e testd 

Fisher exact test %

HarderianGland:Carcinoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Amitage test 

Fisher exact test 

HarderianGland:Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

Liver:HepatocellularAdenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Lifetabletest ' 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

Liver:Hemangiosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test3 

VehicleControl 1 mgkz 25 mgkg 

3/50 (6%) 1/50 (2%) 1/50 (2%) 2/50 (4%) 

7.0% 2.6% 2.6% 4.6% 

540 

2/40 (5 96) 

726 (T) 

1/39 (3%) 

726 (T) 

1/39 (3%) 

405 

1/39 (3X) 

P=O.616 P=0.312N P=0.317N P=0.511N 

P=O.640 P=0.317N P=0.295N P=0.441N 

P=O.623 

P=0.309N P=0.309N P=0.500N 

0150 (0%) 4/50 (8%) 1/50 (2%) 1/50 (2%) 

0.0% 9.7% 2.6% 2.6% 

0140(096) 

-e 705 

2/39 (5%) 

726 (T) 

1/39 (3X) 

726 (T) 

1/39 (3%) 

P=0.467N P=O.O65 P=O.495 P=O.495 

P=0.477N P=O.O63 P=O.495 P=O.495 

P=0.459N 

P=O.O59 P=O.500 P=O.500 

or Carcinoma 

3/50 (6%) 5/50 (10%) 2/50 (4%) 3/50 (6%) 

7.0% 12.1% 5.1% 7.1% 

2/40 ( 5R) 3/39 (8%) 2/39 (5%) 2/39 (5 %) 

540 705 726 (T) 405 

P=0.502N P=O.352 P=0.513N P=O.649 

P=0.484N P=O.352 P=0.499N P=0.619N 

P=0.492N 

P=O.357 P=0.500N P=0.661N 

26/50 (52I) 22/50 (44%) 9/50 (18R) 8/50 (16%) 

59.0% 53.5% 21.3% 19.1% 

22/40 (55%) 20/39 (51R) 6/39 (15%) 6/39 (15%) 

680 705 594 587 

PC0.001N P=0.318N P=0.001N PC0.001N 

PC0.001N P=0.289N PCO.OO1N P<O.OOlN 

PC0.001N 

P=0.274N P<O.OOlN P<0.001N 

0.0% 

0150 (0%) 

0.0% 

0150 (0%) 

9.0% 

4/50 (8%) 

0.0% 

Ol50 (0%) 

0140 (0%) - -

0139 (0%) 

560 

1/39 (3 %) -

0139 (0%) 

P=0.492N - P=O.O61 -

P=0.461N - P=O.O78 -

P=0.480N - P=O.O59 -
Lesions in Male Mice 169 

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-Year Gavage Study 

TABLEC3 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 1 mgkg 5 mag 25 mgkg 

Terminal rate 

Adjusted rate 

Overall rate 

Liver:HepatocellularCarcinoma 

4/40 (10%) 

14.2% 

6/50 (12%) 

6/39 (15%) 

31.0% 

15/50 (30%) 

5/39 (13%) 

14.9% 

6/50 (12 %) 

6/39 (15%) 

17.3% 

7/50 (14%) 

First incidence (days) 405 700 693 622 

Life table test P=0.297N PtO.034 P=O.594 P=O.476 

Logistic regression test P-0.270N P=O.O20 P=O.J77 P=O.468 

Cochran-Armitage test P=0.267N 

Fisher exact test PtO.024 P=0.620N P=O.500 

Liver: .Hepatocellular Adenoma or Hepatocellular Carcinoma 

Overall rate 30150 (60 %) 33\50 (66%) 14/50 (28%) 15/50 (30%) 

Adjusted rate 67.3% 65.2% 32.5 % 35.3% 

Terminal rate 24/40 (60%) 23/39 (59%) 10139 (26%) 12/39 (31 %) 

First incidence (days) 405 680 594 587 

Life table test P=0.003N P=O.328 P=0.004N P=0.007N 

Fisher exact test 

Cochran-Armitage test 

Logistic regression test 

P<O.OOlN 

P-0.001N P~O.348 

P=O.339 

P=O.OOZN 

P=0.001N 

P=0.004N 

P=O.OOZN 

Liver:HepatocellularCarcinoma or Hepatoblastoma 

Overall rate 6/50 (12%) 16\50 (32%) 6/50 (12%) 7/50 (14%) 

Terminal rate 

Adjusted rate 

4/40 (10%) 

14.2% 

7/39 (18%) 

33.1% 

5/39 (13%) 

14.9% 

6/39 (15%) 

17.3% 

First incidence (days) 405 700 693 622 

Fisher exact test 

Cochran-Armitage test 

Logistic regression test 

Life table test 

P=0.230N 

P=0.233N 

P=0.260N 

P=O.O12 

P=O.O22 

P=O.O14 

P=O.577 

P=O.594 

P=0.620N 

P=O.468 

P=O.476 

P=O.500 

e 

Life P=0.003N P=O.328 

First 680 405 

Terminal 24/40 (60%) 23/39 (59%) 

Adjusted 65.2% 67.3% 

Overall 3060 (60%) 33/50 (66%) 

Liver:HepatocellularAdenoma,HepatocellularCarcinoma,orHepatoblastoma 

P=0.004N 

594 

10/39 (26%) 

32.5% 

14/50 (28 46) 

P=0.007N 

587 

12/39 (31 %) 

35.3% 

15/50 (30%) 

ssion 

Fisher 

Cochran-Armitage 

Logistic 

P<0.001N 

P=0.001N 

P10.339 

P=O.348 P=0.002N 

P=O.OOlN 

P=0.004N 

P=0.002N 

Terminal rate 

Adjusted rate 

Overall rate 

Lung:Alveolar/bronchiolarAdenoma 

10/40 (25%) 

28.3% 

12/50 (24%) 

7/39 (18%) 

17.9% 

7/50 (14%) 

7/39 (18%) 

21.7% 

9/50 (1 8 R) 

6/39 (15%) 

15.4% 

6/50 (12%) 

First incidence (days) 683 726 (T) 594 

Life table test P=0.180N 170NP=O. P=0.347N 

726 

P=O.llON 

Logistic regression test P=0.158NP=0.197N P=0.358N P=0.120N 

Cochran-Armitage test P=0.169N 

Fisher exact test P=0.154N P=0.312N P=0.096N 
170 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC3 

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Lung:AlveolarhronchiolarCarcinoma 

Overall rate 3/50 (6%) 4/50 (8 %) 1/50 (2%) 2/50 (4%) 

Adjusted rate 6.8% 9.7% 2.6% 5.1% 

Terminal rate 1140 (3 %) 3/39 (8 %) 1/39 (3%) 2/39 (5%) 

First incidence (days) 680 618 726 (T) 726 (T) 

Life table test P=0.406N P=O.489 P=0.327N P=0.519N 

Logistic regression test P=0.404N P=O.498 P=0.318N P=0.516N 

Cochran-Armitage test P=0.393N 

Fisher exact test P=O.500 P=0.309N P-0.500N 

Lung:AlveolarhronchiolarAdenomaorCarcinoma 

Overall rate 15/50 (30%) 11/50 (22%) 10150 (20%) 8/50 (16%) 

Adjusted rate 33.8% 27.2% 24.2% 20.5% 

Terminal rate 1 1/40 (28 %) 10139 (26%) 8/39 (21 X) 8/39 (21 %) 

First incidence (days) 680 618 594 726 (T) 

Life table test P=0.133N P=0.277N P=0.218N P-0.096N 

Logistic regression test P=0.142N P=0.244N P=0.214N P=0.098N 

Cochran-Armitage test P=0.119N 

Fisher exact test P=0.247N P=0.178N P-0.077N 

PancreaticIslets:Adenoma 

Overall rate 3/50 (6%) 0150 (0%) 1/50 (2%) 1/50 (2%) 

Adjusted rate 7.0% 0.0% 2.3% 2.6% 

Terminal rate 2/40 (5 %) 0139 (0%) 0/39 (0%) 1/39 (3%) 

First incidence (days) 647 - 606 726 (T) 

Life table test P=0.535N P=0.127N P=0.330N P=0.324N 

Logistic regression test P=0.519N P=0.121N P-0.292N P-0.313N 

Cochran-Armitage test P=0.522N 

Fisher exact test P=0.121N P=0.309N P=0.309N 

AllOrgans:Hemangiosarcoma 

Overall rate 1/50 (2%) 2/50 (4%) 5/50 (10%) 0150 (0%) 

Adjusted rate 2.4% 4.3% 11.2% 0.0% 

Terminal rate 0/40 (0%) 0139 (0%) 1/39 (3%) 0139 (0%) 

First incidence (days) 725 405 560 - Life table test P=0.200N P=O.499 P=O.O99 P=0.510N 

Logistic regression test P=0.132N P=O.433 P=O.120 P=0.511N 

Cochran-Armitage test P=0.187N 

,Fisher exact test P=O.500 P=O.102 P=0.500N 

All Organs:Hemangioma or Hemangiosarcoma 

Overall rate 2/50 (4%) 2/50 (4%) 5/50 (10%) 0150 (0%) 

Adjusted rate 4.6% 4.3% 11.2% 0.0% 

Terminal rate 0140 (0%) 0139 (0%) 1/39 (3%) 0/39 (0%) 

First incidence (days) 680 405 560 -

Life table test P=0.149N P=O.687 P=O.203 P=0.255N 

Logistic regression test P=0.092N P10.650 P=O.331 P=0.240N 

Cochran-Armitage test P=0.135N 

Fisher exact test P=0.691N P-0.218 P=0.247N 
 

Lesions in Male Mice 171 

TABLEC3 

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 1 mgflrg 

Life table test 

First incidence (days) 

Terminal rate 

Adjusted rate 

Overall rate 

All Organs:HistiocyticSarcoma 

P=0.589N 

447 

2\40 (5%) 

6.9% 

3/50 (6%) 

0139 (0%) 

0.0% 

0150 (0%) 

P=0.126N 

- 0139 (0%) 

0.0% 

0/50 (0%) 

P=0.129N 

- P=0.322N 

707 

0139 (0%) 

2.5% 

1/50 (2%) 

Fisher exact test 

Cochran-Armitage test 

Logistic regression test 

P=0.579N 

P=0.566N 

P=0.121N 

P=0.138N 

P=0.121N 

P=0.092N P=0.275N 

P=0.309N 

ion 

Fisher exact test 

Cochran-Annitage 

Logistic 

Life 

First 

Terminal 

Adjusted 

Overall 

All Organs:MalignantLymphoma(Lymphocytic 

P=O.600 

P=O.577 

P=O.588 

680 

3/40 (8 X) 

9.5% 

4/50 (8 %) 

or Mixed) 

P=0.337N 

P=0.351N 

726 (T) 

2/39 (5%) 

5.1% 

2/50 (4%) 

P=0.339N 

P=0.361N 

P=0.358N 

707 

1/39 (3 %) 

5.0% 

2/50 (4%) 

P=0.339N 

P=0.526N 

P=0.519N 

705 

2/39 (5%) 

7.4% 

3/50 (6%) 

P=OSOON 

Fisher exact test 

Cochran-Armitage test 

Logistic regression test 

Life table test 

First incidence (days) 

Terminal rate 

Adjusted rate 

Overall rate 

All Organs:BenignNeoplasms 

P=0.002N 

P=0.003N 

P=O.WN 

540 

30140 (75 %) 

78.2% 

36/50 (72%) 

P=0.063N 

P=0.104N 

705 

26/39 (67 %) 

68.2% 

28/50 (56%) 

P=0.072N 

P=0.004N 

P=O.OlON 

520 

15/39 (38 X) 

46.6% 

21/50 (42%) 

P=0.002N 

P<0.001N 

P=0.001N 

405 

15/39 (38%) 

42.3% 

18/50 (36%) 

P<0.001N 

Fisher exact test 

Cochran-Armitage test 

Logistic regression test 

Life table test 

First incidence (days) 

Terminal rate 

Adjusted rate 

Overall rate 

AllOrgans:MalignantNeoplasms 

P=0.136N 

P=0.130N 

P=0.189N 

447 

1 1/40 (28 %) 

36.8% 

17/50 (34%) 

P10.108 

P=O.105 

405 

14/39 (36%) 

50.0% 

25/50 (50%) 

P=O.O78 

P=0.524N 

P=0.546N 

560 

10139 (26%) 

35.5% 

16\50 (32%) 

P=0.500N 

P=0.423N 

P=0.464N 

405 

11/39 (28 %) 

34.6% 

15/50 (30%) 

P=0.415N 
172 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEC3 

Statistical Analysis of Primary Neoplasms in Male Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) . Vehicle Control 1 mglkg 5 mgkg 25 mgkg 

All Organs: Benign or Malignant Neoplasms 

Overall rate 43/50 (86%) 41/50 (82%) 31/50 (62%) 29/50 (58%) 

Adjusted rate 87.8% 82.0% 66.0% 62.8% 

Terminal rate 34/40 (85%) 30/39 (77 X) 23/39 (59%) 22/39 (56%) 

First incidence (days) 447 405 520 405 

Life table test P=0.019N P=0.490N P=0.045N P=0.018N 

Logistic regression test P=0.002N P=0.388N P=O.OlON P=0.002N 

Cochran-Annitage test P=0.002N 

Fisher exact test P=0.393N P=0.006N P=0.002N 

(T)Terminal sacrifice 

a Number of neoplasm-bearing animalslnumber of animals examined. Denominator is number of animals examined microscopically for liver, lung, and 

pancreatic islets; for other tissues, denominator is number of animals necropsied. 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence at terminal kill 

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 priorto 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 vend or a lower incidence in a dose group is indicated by N. 

e Not applicable; no neoplasmsinanimalgroup 
Lesions in Male Mice 173 

TABLEC4 

Historical Incidence of Hepatocellular Neoplasms in Male B6C3Fl Mice Administered Water by Gavagea 

IncidenceinControls 

Adenoma Carcinomaor Adenoma Carcinoma 

Overall Historical Incidence 

Total 401315 (12.7%) 39/315 (12.4%) 741315 (23.5%) 

Standard deviation 5.2% 6.1% 7.2% 

Range 4%-18% 6%-24% 1446-3646 

a Data as of 17 June 1994 
174 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC5 

Summary of the Incidence of Nonneoplastic Lesions inMale Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydratea 

Disposition Summary 

Animals initially in study 

15-Month interim evaluationb 

Early deaths 

Accidental deaths 

Moribund 

Natural deaths 

Survivors 

Terminal sacrifice 

Animals examined microscopically 

15-Month Interim Evaluation 

AlimentarySystem 

Liver 

Clear cell focus 

Eosinophilic focus 

Fatty change 

Mixed cell focus 

Bile duct, hyperplasia 

Hepatocyte, hypertrophy 

Mesentery 

Fat, inflammation, chronic active 

Pancreas 

Atrophy 

Stomach, forestomach 

Hyperplasia, focal 

Tooth 

Dysplasia 

EndocrineSystem 

Adrenal cortex 

Hyperplasia 

Capsule, hyperplasia, adenomatous 

Islets, pancreatic 

Hyperplasia 

Pituitary gland 

cyst 

GenitalSystem 

Epididymis 

Inflammation, chronic active 

Preputial gland 

Duct, ectasia 

Vehicle Control 

70 

20 

4 

6 

40 

60 

(10) 

1 (10%) 

1 (10%) 

2 (20%) 

1 (10%) 

1 (10%) 

(10) 

4 (40%) 

1 (10%) 

(10) 

3 (30%) 

(10) 

1 (10%) 

1 mgk 

70 

20 

7 

4 

39 

60 

(10) 

1 (10%) 

2 (20%) 

4 (40%) 

1 (10%) 

1 (10%) 

(10) 

1 (10%) 

(10) 

7 (70%) 

(9) 

5 mgk 

70 

20 

2 

4 

5 

39 

60 

2 (20%) 

(10) 

2 (20%) 

1 (10%) 

(10) 

4 (40%) 

(9) 

1 (11%) 

25 mglkg 

70 

20 

2 

7 

2 

39 

60 

(10) 

2 (20%) 

3(30%) 

(10) 

(9) 

a Number of animals examined microscopically at the site and the number of animals with lesion 

Ten animals per group received ophthalmic examinations and were discarded without further evaluation 
Lesions in Male Mice 175 

TABLEC5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

15-Month Interim Evaluation (continued) 

Musculoskeletal System 

Skeletal muscle (1) 

Degeneration 1 (100%) 

Respiratory System 

Lung (10) 

Alveolar epithelium, hyperplasia 

Special Senses System 

EY e 

Degeneration 

Harderian gland (4) (2)

Inflammation, chronic active 

Urinary System 

Kidney 

Nephropathy 

Systems Examined With No Lesions Observed 

Cardiovascular System 

General Body System 

HematopoieticSystem 

Integumentary System 

Nervous System 

2-Year Study 

Alimentary System 

Esophagus 

cyst 

Periesophageal tissue, inflammation, 

suppurative 1 (2%) 

Gallbladder (49) 

Inflammation, chronic active 

Intestine small, jejunum 

Hyperplasia, lymphoid 

Ulcer 

Intestine small, ileum (49) (49)

Amyloid deposition 
176 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEC5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 

&Year Study (continued) 

Alimentary System (continued) 

Liver (50) 

Amyloid deposition 

Basophilic focus 3(6%) 

Clear cell focus 12 (24%) 

Eosinophilic focus 21 (42%) 

Fatty change 1 (2%) 

Hematopoietic cell proliferation 2 (4%) 

Inflammation, chronic active 1(2%) 

Mixed cell focus 5 (10%) 

Necrosis 1(2%) 

Mesentery (4) 

Infiltration cellular, lymphocyte 

Fat, inflammation, chronic active 

Fat, necrosis 

Pancreas 

Atrophy 

Hyperplasia 

Necrosis 

Duct, cyst 

Stomach, forestomach 

Hyperplasia, focal 

Stomach, glandular 

Inflammation, chronic active, focal 

Tooth (14) 

Dysplasia 13 (93%) 

Cardiovascular System 

Blood vessel (50) 

Aorta, inflammation, chronic active 

Heart (50) 

Inflammation, chronic active 

Mineralization 2(4%) 

Artery, inflammation, chronic active 

Artery, mineralization 

Endocrine System 

Adrenal cortex (50) 

Accessory adrenal cortical nodule 1(2%) 

Hyperplasia 23 (46%) 

Capsule, hyperplasia, adenomatous 7 (14%) 

Adrenal medulla (49) 

Hyperplasia 

Inflammation, chronic active 

Islets, pancreatic 

Hyperplasia 

Parathyroid gland 

Hyperplasia 

4 (8%) 

4 (8%) 

12 (24%) 

1 (2%) 

7 (14%) 

2 (4%) 

(3) 

2 (67%) 

(50) 

1 (2%) 

1 (2%) 

1 (2%) 

(50) 

2(4%) 

(50) 

1 (2%) 

(4) 

3 (75%) 

(50) 

(50) 

2(4%) 

(50) 

17 (34%) 

2(4%) 

(45) 

3 (7%) 

3(6%) 

2 (4%) 

7 (14%) 

1 (2%) 

7 (14%) 

1 (2%) 

(3) 

1 (33%) 

1 (33%) 

1 (33%) 

(50) 

1 (2%) 

(4) 

2 (50%) 

(50) 

(50) 

1 (2%) 

1 (2%) 

25 mgkg 

(50) 

1 (2%) 

1 (50%) 

(2) 

1 (50%) 

(50) 

1 (2%) 

(50) 

1 (2%) 
Lesions in Male Mice 177 

TABLEC5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

&Year Study (continued) 

Endocrine System (continued) 

Pituitary gland 

Pars distalis, hyperplasia 

Pars intermedia, hyperplasia 

Thyroid gland 

Follicular cell, hyperplasia 

General Body System 

None 

Genital System 

Epididymis 

Granuloma sperm 

Inflammation, chronic active 

Artery, inflammation, chronic active 

Preputial gland 

Inflammation, chronic active 

Duct, ectasia 

Prostate 

Inflammation, chronic active 

Artery, inflammation, chronic active 

Seminal vesicle 

Inflammation, chronic active 

Testes 

Atrophy 

' Interstitial cell, hyperplasia 

Hematopoietic System 

Bone marrow 

Thrombosis 

Erythroid cell, hyperplasia 

Myeloid cell, hyperplasia 

Lymph node 

Lumbar, hyperplasia, lymphoid 

Mediastinal, hyperplasia, lymphoid 

Lymph node, mandibular 

Hyperplasia, lymphoid 

Lymph node, mesenteric 

Hematopoietic cell proliferation 

Hyperplasia, lymphoid 

Hyperplasia, plasma cell 

Inflammation, chronic 'active 

Spleen 

Amyloid deposition 

Angiectasis 

Depletion lymphoid 

Hematopoietic cell proliferation 

Hyperplasia, lymphoid 

Thymus 

Atrophy 

(48) 

1 (2%) 

(50) 

13 (26%) 

(50) 

1 (2%) 

(50) 

1 (2%) 

47 (94%) 

(49) 

(48) 

2 (4%) 

1 (2%) 

1 (2%) 

8 (16%) 

1 (2%) 

(42) 

6 (14%) 

(47) 

(50) 

15 (30%) 

(50) 

2(4%) 

4 (8%) 

4 (8%) 

1(2%) 

(50) 

1 (2%) 

12 (24%) 

(44)

12 (27%) 

(46) 

1 (2%) 

(50) 

13 (26%) 

5(10%) 

(2) 

1 (2%) 

14 (28%) 

(40) 

4 (10%) 

(44) 

1 (2%) 

(50) 

5 (10%) 

1 (2%) 

(50) 

3 (6%) 

45 (90%) 

(50) 

2(4%) 

1 (2%) 

(46) 

1 (2%) 

(47) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

3 (6%) 

12 (24%) 

(40)

7 (18%) 
178 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

of Scopolamine Hydrobromide Trihydrate (continued) 

Summary of the Incidence of Nonneoplastic Lesionsin Male Mice in the 2-Year Gavage Study 

TABLEC5 

Skin 

IntegumentarySystem 

%Year Study (continued) 

Subcutaneous tissue, inflammation, chronic 

Ulcer 

Inflammation, chronic active 

active 

Subcutaneous tissue, lymphangiectasis 1 (2%) 

None 

MusculoskeletalSystem 

Brain 

NervousSystem 

Neuron, necrosis 

Artery, inflammation, chronic active 

Hemorrhage, acute 

1 (2%) 

Lung 

RespiratorySystem 

Alveolar epithelium, hyperplasia 

Pigmentation, hemosiderin 

Inflammation, suppurative 

Inflammation, chronic active 

Hemorrhage, acute 

Embolus 

Peritracheal tissue, hemorrhage, acute 

Inflammation, suppurative 

Hemorrhage, acute 

Nose 

Trachea 

(50) 

1 (2%) 

8 (16%) 

1 (2%) 

(50) 

(50) 

(50) 

3(6%) 

1 (2%) 

(50) 

1 (2%) 

(50) 

(50) 

1 (2%) 

2 (4%) 

1 (2%) 

2 (4%) 

1(2%) 

(50) 

(50) 

1 (2%) 

(50) 

1 (2%) 

1 (2%) 

1 (2%) 

(50) 

(50) 

2 (4%) 

EY e 

Special SensesSystem 

Hyperplasia 

Harderian gland 

Cornea, necrosis 

Cornea, inflammation, chronic active 

(16) 

1 (6%) 

(3) 

1(33%) 

1(33%) 

(18) 

(1) 

1(100%) 

(19) 

(2) 

1 (50%) 

(13) 
Lesions in Male Mice 179 

TABLEC5 

Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Urinary System 

2-YeUr study (continued) 

Kidney 

cyst 

Hydronephrosis 

Infarct 

Inflammation, chronic active 2 (4%) 

Necrosis 1 (2%) 

Nephropathy 48 (96%) 46 (92%) 

Artery, inflammation, chronic active 

Glomerulus, amyloid deposition 

Renal tubule, hyperplasia 

Urinary bladder 

Inflammation, chronic active 

(50) 

2(4%) 

1 (2%) 

2 (4%) 

42 (84%) 

2(4%) 

(50) 

2(4%) 

(50) 

3 (6%) 

1 (2%) 

2(4%) 

1 (2%) 

37 (74%) 

1 (2%) 

(49) 

3 (6%) 
180 Scopolamine Hydrobromide Trihydrate,NTP TR 445 
181 

APPENDIX D 

SUMMARY OF LESIONS IN FEMALE MICE 

IN THE 2-YEAR GAVAGE STUDY 

OF SCOPOLAMINE HYDROBROMIDE TRIHYDRATE 

TABLEDl Summary of the Incidence of Neoplasms in Female Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate . . . ... . . . . . 183 

TABLED2 Individual Animal Tumor Pathology of Female Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate .. . . . .... . . 188 

TABLED3 Statistical Analysis of Primary Neoplasms in Female Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate .. . . . . . . .. . 206 

TABLED4 Historical Incidence of Hepatocellular Neoplasms 

in Female B6C3Fl MiceAdministered Water by Gavage . . . . . . .. . . . . . . . . . . . . 210 

TABLED5 Summary of the Incidence of Nonneoplastic Lesions in Female Mice 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate . . . . . . . . . . . 211 
182 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

c 
Lesions in Female Mice 183 

of Scopolamine Hydrobromide Trihydratea 

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Gavage Study 

TABLEDl 

Early deaths 

15-Month interim evaluadbnb 

Animals initially in study 

Disposition Summary 

Natural deaths 

Moribund 

Accidental deaths 

Terminal sacrifice 

Survivors 

19 

70 

7 

9 

2 

33 

20 

70 

3 

11 

36 

20 

70 

5 

7 

1 

37 

19 

70 

6 

7 

38 

Animals examined microscopically 61 60 60 61 

Liver 

Alimentary System 

15-Month Interim Evaluation 

Hepatocellular adenoma 

(10) 

1 (10%) 

(10) 

Thyroid gland 

Endocrine System 

Follicular cell, adenoma 

Cystadenoma 

Ovaly 

Genital System 

Urinary System 

Special Senses System 

Respiratory System 

Nervous System 

Musculoskeletal System 

Integumentary System 

Hematopoietic System 

General Body System 

Cardiovascular System 

Systems Examined With No Neoplasms Observed 
184 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEDl 

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

2-Year Study 

AlimentarySystem 

Gallbladder 

Intestine large, rectum 

Anus, sarcoma 

Intestine small, duodenum 

Hemangiosarcoma 

Intestine small, jejunum 

Carcinoma 

Liver 

Hemangiosarcoma 

Hemangiosarcoma, multiple 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Hepatocholangiocarcinoma 

Histiocytic sarcoma 

Bile duct, carcinoma 

Mesentery 

Carcinoma, metastatic, liver 

Histiocytic sarcoma 

Pancreas 

Carcinoma, metastatic, liver 

Histiocytic sarcoma 

Salivary glands 

Stomach, forestomach 

Mast cell tumor benign 

Squamous cell papilloma 

Stomach, glandular 

Cardiovascular System 

Heart 

Carcinoma, metastatic, liver 

Hepatocholangiocarcinoma,metastatic, liver 

EndocrineSystem 

Adrenal cortex 

Capsule, adenoma 

Adrenal medulla 

Pheochromocytoma malignant 

Pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Pituitaly gland 

Pars distalis, adenoma 

Pars intennedia, adenoma 

Thyroid gland 

Follicular cell, adenoma 

1 (2%) 

7 (14%) 

1 (2%) 

10 (20%) 

5 (10%) 

1 (2%) 

1 (2%) 

1 (2%) 

(7) 

1 (14%) 

(51) 

1 (2%) 

(51) 

1 (2%) 

1 (2%) 

(51) 

1 (2%) 

(50) 

4 (8%) 

(51) 

3 (6%) 

(49) 

(49) 

(50) 

1 (2%) 

5 (10%) 5 (10%) 4 (8%) 

1 (2%) 3 (6%) 

13 (26%) 9 (18%) 4 (8%) 

5 (10%) 2(4%) 

2 (4%) 1 (2%) 

(50) 
Lesions in Female Mice 185 

TABLEDl 

Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

%Year Study (continued) 

General Body System 

None 

Genital System 

Ovary 

Cystadenoma 

Fibroma 

Hemangiosarcoma 

Luteoma 

uterus 

Adenoma 

Hemangioma 

Hemangiosarcoma 

Hemangiosarcoma, metastatic, ovary 

Polyp stromal 

Sarcoma stromal 

HematopoieticSystem 

Bone marrow 

Hemangiosarcoma 

Histiocytic sarcoma 

Lymph node 

Bronchial, carcinoma, metastatic, harderian 

gland 

Bronchial, carcinoma, metastatic. liver 

Bronchial, hepatocholangiocarcinoma, 

metastatic, liver 

Mediastinal, carcinoma, metastatic, liver 

Mediastinal, hepatocholangiocarcinoma, 

metastatic, liver 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma 

Histiocytic sarcoma 

Thymus 

Hepatocholangiocarcinoma, metastatic, liver 

Histiocytic sarcoma 

Mast cell tumor malignant 

Mediastinum,rhabdomyosarcoma,metastatic, 

skeletal muscle 1 (2%) 

Integumentary System 

Skin (51) (50) (50) (51)

Squamous cell carcinoma 1 (2%) 

Subcutaneous tissue, fibrosarcoma 1 (2%) 1 (2%) 1 (2%) 

Subcutaneous tissue, hemangiosarcoma 1 (2%) 
186 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEDl 

Summary of the Incidence of Neoplasms in Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

%Year Study (continued) 

MusculoskeletalSystem 

Bone 

Hepatocholangiocarcinoma, metastatic, liver 

Skeletal muscle 

Hepatocholangiocarcinoma, metastatic, liver 

Rhabdomyosarcoma 

NervousSystem 

None 

RespiratorySystem 

Lung 

Alveolarlbronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, harderian gland 

Carcinoma, metastatic, liver 

Hemangiosarcoma, metastatic, ovary 

Hepatocellular carcinoma, metastatic, liver 

Hepatocholangiocarcinoma, metastatic, liver 

Histiocytic sarcoma 

Special Senses System 

Harderian gland 

Adenoma 

Carcinoma 

UrinarySystem 

Kidney 

Carcinoma, metastatic, liver 

Hepatocholangiocarcinoma, metastatic, liver 

Histiocytic sarcoma 

Urinary bladder 

SystemicLesions 

Multiple organs' 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

Lymphoma malignant undifferentiated cell 

1 (2%) 

(1) 

1 (100%) 

(51) 

3 (6%) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

1(2%) 

1 (2%) 

(17) 

1 (6%) 

2 (12%) 

(51) 

1 (2%) 

1 (2%) 

(50) 

(51) 

1(2%) 

2 (4%) 

7 (14%) 

(50) 

4 (8%) 

3 (6%) 

2 (4%) 

(14) 

3 (21%) 

1(7%) 

(50) 

1 (2%) 

(48) 

(50) 

2 (4%) 

3 (6%) 

5 (10%) 

(50) 

1(2%) 

1 (2%) 

1 (2%) 

5 (10%) 

(15) 

2 (13%) 

1 (7%) 

(50) 

2 (4%) 

6 (12%) 

1 (2%) 

(5 1) 

2 (4%) 

1(2%) 

1 (2%) 

(13) 

1 (8%) 

(51) 

1 (2%) 

4 (8%) 

3 (6%) 
Lesions in Female Mice 187 

TABLEDl 

Summary of the Incidence of Neoplasms in Female Mice in the2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Neoplasm Summary 

Total animals with primary neoplasmsd 

15-Month interim evaluation 1 1 

2-Year study 40 36 35 29 

Total primary neoplasms 

15-Month interim evaluation 2 1 

2-Year study 62 58 45 35 

Total animals with benign neoplasms 

15-Month interim evaluation 1 1 

2-Year study 24 25 21 18 

Total benign neoplasms 

15-Month interim evaluation 2 1 

2-Year study 31 34 22 19 

Total animals with malignant neoplasms 

2-Year study 26 22 21 14 

Total malignant neoplasms 

2-Year study 31 24 23 16 

Total animals with metastatic neoplasms 

2-Year study 5 2 7 1 

Total metastatic neoplasms 

2-Year study 19 2 8 1 

a Number of animals examined microscopically at the site and the number of animals with neoplasm 

Nine or ten animals per group received ophthalmic examinations and were discarded without further evaluation 

Number of animals with any tissue examined microscopically 

Primary neoplasms: all neoplasms except metastatic neoplasms 
188 Scopolamine Hydrobromide Trihydrate,TR 445 

TABLED2 

Individual AnimalTumor Pathology of Female Mice in the 2-Year Gavage.Study 

of ScopolamineHydrobromideTrihydrate:Vehicle Control 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Carcinoma 

Intestine small, ileum 

Liver 

Hemangiosarcoma, multiple 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Hepatocholangiocarcinoma 

Histiocytic sarcoma 

Bile duct, carcinoma 

Mesentery 

Carcinoma, metastatic, liver 

Pancreas 

Carcinoma, metastatic, liver 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Carcinoma, metastatic, liver 

Hepatocholangiocarcinoma, metastatic, 

liver 

EndocrineSystem 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma malignant 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

+: Tissue examined microscopically 

A: Autolysis precludes examination 

0345556666666777777777777 

0791890111167001122222222 

7321144011819792706668888 

4343333333333333333333333 

0806977569994974587894445 

0847353155602467508786894 

......................... 

......................... 

......................... 

......................... 

++++++++++++M+M.M+++++++++ 

......................... 

......................... 

++++++++++++M++++++++++++ 

.......................... X 

X X 

X 

X x x 

X X 

X 

X 

X 

+ ++ + + 

X ......................... X ......................... 

......................... 

......................... 

......................... 

......................... X 

X 

......................... 

......................... X ......................... 

M++++++++++++++++++++++++ 

+++++++M+++++++++++++++++ 

X ......................... X 

M: Missing tissue X: Lesion present 

I: Insufficient tissue Blank: Not examined 
Lesions in Female Mice 189 

TABLED2 

Individual Animal Tumor Pathologyof Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: Vehicle Control (continued) 

Number of DaysonStudy 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Carcinoma 

Intestine small, ileum 

Liver 

Hemangiosarcoma,multiple 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Hepatocholangiocarcinoma 

Histiocytic sarcoma 

Bile duct, carcinoma 

Mesentery 

Carcinoma, metastatic. liver 

Pancreas 

Carcinoma.metastatic,liver 

Salivary glands 

Stomach,forestomach 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Carcinoma, metastatic, liver 

Hepatocholangiocarcinoma,metastatic, 

liver 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Pheochromocytoma malignant 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

77777777777777777777777777 

22222222222233333333333333 

88888888888800000000000000 

33333333344433333333334444 Total 

55667789900044456688880001 Tissues/ 

79282762935934534612392670 Tumors 

.......................... 51 

.......................... 51 

.......................... 51 

.......................... 51 

.......................... 48 

.......................... 51 

.......................... 51 

X 1 

.......................... 50 

............................ 51 

1 

X xx X X 7 

1 

X X X xx x X 10 

X x x 5 

1 

1 

1 

+ + 7 

1 

.......................... 51 

1 

.......................... 51 

.......................... 51 

.......................... 51 

.......................... 51 

.......................... 51 

1 

1 

.......................... 51 

.......................... 51 

1 

.......................... 51 

++++++++++M++++++++++++++M 48 

.......................... 50 

X x x 4 

.......................... 51 

X X 3 
Scopolamine Hydrobromide Trihydrate,TR 445 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

Number of Days on Study 

Carcass ID Number 

Genital System 

Clitoral gland 

Ovary 

Cystadenoma 

Hemangiosarcoma 

uterus 

Adenoma 

Hemangiosarcoma 

Hemangiosarcoma, metastatic, ovary 

Polyp stromal 

Sarcoma stromal 

HematopoieticSystem 

Bone marrow 

Histiocytic sarcoma 

Lymph node 

Bronchial, carcinoma, metastatic, 

liver 

Bronchial, hepatocholangiocarcinoma, 

metastatic, liver 

Mediastinal, carcinoma, metastatic, 

liver 

Mediastinal, 

hepatocholangiocarcinoma, 

metastatic, liver 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma 

Histiocytic sarcoma 

Thymus 

Hepatocholangiocarcinoma, metastatic, 

liver 

Histiocytic sarcoma 

Integumentary System 

Mammary gland 

Skin 

Subcutaneous tissue, fibrosarcoma 

MusculoskeletalSystem 

Bone 

Hepatocholangiocarcinoma, metastatic, 

liver 

Skeletal muscle 

Hepatocholangiocarcinoma, metastatic, 

liver 

~ ~ ~~ ~ ~ ~~ ~~~ ~ ~ ~ 

0345556666666777777777777 

0791890111167001122222222 

7321144011819792706668888 

4343333333333333333333333 

0806977569994974587894445 

0847353155602467508786894 

........................ 

......................... X X X 

X 

......................... 

X 

......................... X 

+ + ++ + 

X 

X 

X 

X 

X 

+++++++++++M+++++++++++++ 

++M++++++++++++++++++++++ 

......................... 

X 

+++++++++++++++M++M++++++ 

X 

X 

......................... 

......................... X 

......................... 

X 

+ 

X 
Lesions in Female Mice 191 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: Vehicle Control (continued) 

Number of Days on Study 

Carcass ID Number 

Genital System 

Clitoral gland 

Ovary 

Cystadenoma 

Hemangiosarcoma 

Uterus 

Adenoma 

Hemangiosarcoma 

Hemangiosarcoma, metastatic, ovary 

Polyp stromal 

Sarcoma stromal 

Hematopoietic System 

Bone marrow 

Histiocytic sarcoma 

Lymph node 

Bronchial, carcinoma, metastatic, 

liver 

Bronchial, hepatocholangiocarcinoma, 

metastatic, liver 

Mediastinal, carcinoma, metastatic, 

liver 

Mediastinal, 

hepatocholangiocarcinoma, 

metastatic, liver 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Hemangiosarcoma 

Histiocytic sarcoma 

Thymus 

Hepatocholangiocarcinoma, metastatic, 

liver 

Histiocytic sarcoma 

Integumentary System 

Mammary gland 

Skin 

Subcutaneous tissue, fibrosarcoma 

Musculoskeletal System 

Bone 

Hepatocholangiocarcinoma, metastatic, 

liver 

Skeletal muscle 

Hepatocholangiocarcinoma, metastatic, 

liver 

77777777777777777777777777 

22222222222233333333333333 

88888888888800000000000000 

33333333344433333333334444 Total 

55667789900044456688880001 Tissues/ 

79282762935934534612392670 Tumors 

+++++++++++++++++++++M++++ 48 

.......................... ' 51 

3 

1 

.......................... 51 

X 1 

X 1 

1 

X 1 

X 1 

.......................... 51 

1 

+ 6 

1 

1 

1 

1 

1 

.......................... 50 

.......................... 50 

.......................... 51 

X 1 

1 

M+++++++++++++++++++++++++ 48 

1 

1 

.......................... 51 

.......................... 51 

1 

.......................... 51 

1 

1 

1 
192 Scopolamine Hydrobromide Trihydrate, TR 445 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Gavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

Number of Dayson Study 

Carcass ID Number 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Alveolar/bronchiolar adenoma 

Alveolarlbronchiolar carcinoma 

Carcinoma, metastatic, harderian 

gland 

Carcinoma, metastatic. liver 

Hemangiosarcoma, metastatic, ovary 

Hepatocellular carcinoma, metastatic, 

liver 

Hepatocholangiocarcinoma, metastatic, 

liver 

Histiocytic sarcoma 

Nose 

Trachea 

Special Senses System 

Eye

Harderian gland 

Adenoma 

Carcinoma 

UrinarySystem 

Kidney 

Carcinoma, metastatic, liver 

Hepatocholangiocarcinoma, metastatic, 

liver 

Urinary bladder 

SystemicLesions 

Multiple organs 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

03455566666.66777777777777777 

0791890111167001122222222 

7321144011819792706668888 

4343333333333333333333333 

0806971569994974587894445 

0847353155602467508186894 

......................... + + 

+ + 

......................... X X 

X 

X 

X 

X 

X 

X ......................... 

......................... 

+ 

+M++M+M+MMMMM++MMM+M+M+MM 

X X 

......................... X 

X 

++++++++++++++++++++M++++ 

......................... X 

X 

x x X 
Lesions in Female Mice 193 

TABLED2 

Individual Animal Tumor Pathologyof Female Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate:VehicleControl (continued) 

Number of Days on Study 

Carcass ID Number 

NervousSystem 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, harderian 

gland 

Carcinoma, metastatic, liver 

Hemangiosarcoma, metastatic, ovary 

Hepatocellular carcinoma, metastatic, 

liver 

Hepatocholangiocarcinoma, metastatic, 

liver 

Histiocytic sarcoma 

Nose 

Trachea 

Special Senses System 

EY e Harderian gland 

Adenoma 

Carcinoma 

UrinarySystem 

Kidney 

Carcinoma, metastatic, liver 

Hepatocholangiocarcinoma, metastatic, 

liver 

Urinary bladder 

SystemicLesions 

Multiple organs 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

77777777777777777777777777 

22222222222233333333333333 

88888888888800000000000000 

33333333344433333333334444 Total 

55667789900044456688880001 Tissues/ 

79282762935934534612392670 Tumors 

.......................... 51 

2 

1 

.......................... 51 

X 3 

X 1 

1 

1 

1 

.......................... 51 

.......................... 51 

+ 2 

M+MMMMM+M+MM+M++MMMM+MMMMM 17 

X 1 

2 

.......................... 51 

1 

1 

.......................... 50 

.......................... 51 

1 

X 2 

xx X X 7 
194 Scopolamine Hydrobromide Trihydrate, TR 445 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 1 mg/kg 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hemangiosarcoma 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Histiocytic sarcoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Capsule, adenoma 

Adrenal medulla 

Pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Pars intermedia, adenoma 

Thyroid gland 

~~~~~~ 

General Body System 

None 

Genital System 

Clitoral gland 

Ovary 

Fibroma 

Uterus 

Sarcoma stromal 

4445555666677777777777777 

1483699378900022222222222 

5822204060458866666888888 

5555544454554445555444444 

2131397847207991223788889 

6197451208742186082635893 

......................... 

......................... 

......................... 

......................... 

+++++++++++++++++++++++M+ 

......................... 

++++++++++++++M++++++++++ 

++++++++++++++++++M++++++ 

......................... X 

X 

X 

xx X X 

X x x 

X X 

+ + + ......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... xx ......................... X 

M+++++++M+++++++M+M+++++M 

++++++++M++++++++++++++++ 

xx X 

X 

......................... 

++M++++++++++++++++++++++ 

......................... 

......................... X 
Lesions in Female Mice 195 

TABLED2 

Individual Animal Tumor Pathologyof Female Mice in the 2-Yar Gavage Study 

of ScopolamineHydrobromideTrihydrate: 1 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hemangiosarcoma 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Histiocytic sarcoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

EndocrineSystem 

Adrenal cortex 

Capsule, adenoma 

Adrenal medulla 

Pheochromocytoma benign 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Pars intermedia, adenoma 

Thyroid gland 

General Body System 

None 

GenitalSystem 

Clitoral gland 

Ovary 

Fibroma 

Uterus 

Sarcoma stromal 

7777777777777777777777'777 

2222222222233333333333333 

8888888888800000000000000 

4455555555544445555555555 

9900111112377780000112233 

6778034894534571569251236 

......................... 

......................... 

......................... 

......................... 

......................... 

+++++++++++++M+++++++++++ 

......................... 

......................... 

......................... 

X X X X 

xx x x x xxx X 

X X 

+++ + + + ......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... X 

+++++++++M+++++++++++++++ 

......................... 

+++M++++MM+++++++++++M+++ 

M+++++++M++++++++++++++++ 

......................... 

++M+++++M+++M++++++++++++ 

++++++++++++++++++++++M++ 

X ......................... 

Total 

Tissues/ 

Tumors 

50 

50 

50 

50 

' 49 

49 

49 

49 

50 

1 

5 

1 

13 

5 

2 

9 

50 

50 

50 

50 

50 

50 

50 

1 

49 

2 

50 

1 

41 

47 

3 

1% 

50 

46 

49 

1 

50 

1 
196 Scopolamine Hydrobromide Trihydrate, TR 445 

TABLED2 

Individual Animal Tumor Pathologyof Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 1mglkg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Histiocytic sarcoma 

Thymus 

IntegumentarySystem 

Mammary gland 

Skin 

Subcutaneous tissue, fibrosarcoma 

Subcutaneous tissue, hemangiosarcoma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

RespiratorySystem 

Lung 

Alveoladbronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Trachea 

Special Senses System 

EY e Harderian gland 

Adenoma 

Carcinoma 

Urinary System 

Kidney 

Histiocytic sarcoma 

Urinary bladder 

SystemicLesions 

Multiple organs 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

4445555666611111111111111 

1483699318900022222222222 

5822204060,458866666888888 

5555544454554445555444444 

2131391841201991223188889 

6191451208142186082635893 

......................... + + + 

+++++++++++++++++M+++++++ 

+M++++++++++++M++++++++++ 

......................... A 

MM++++MM+M+++++++M+++++++ 

......................... 

......................... X 

X 

......................... 

......................... + + 

+ + 

......................... X 

X X 

......................... 

......................... 

+ 

MM+MMMMMMMMMMMM+MMMMM+M+M 

X 

......................... X 

+++++++++++++++++M+++++++ 

......................... X X 

X 

X xx 
Lesions in Female Mice 197 

TABLED2 

Individual Animal Tumor Pathologyof Female Mice in the 2-YearGavage Study 

of ScopolamineHydrobromideTrihydrate: 1 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Bone marrow 

Lymph node 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Histiocytic sarcoma 

Thymus 

Integumentary System 

Mammary gland 

Skin 

Subcutaneous tissue, fibrosarcoma 

Subcutaneous tissue, hemangiosarcoma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Trachea 

Special Senses System 

Eye

Harderian gland 

Adenoma 

Carcinoma 

Urinary System 

Kidney 

Histiocytic sarcoma 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

7777777777777777777777777 

2222222222233333333333333 

8888888888800000000000000 

4455555555544445555555555 Total 

9900111112377780000112233 Tissues/ 

6778034894534571569251236 Tumors 

......................... 50 

+ + 5 

++M+++++++M++++++M+++++++ 46 

++++M++++++++++++++++++++ 47 

......................... 50 

1 

++++++++++M++++++++++++++ 43 

+++++++M+++++++++++++++++ 49 

......................... 50 

1 

1 

......................... 50 

......................... 50 

2 

2 

......................... 50 

xx X 4 

X 3 

X X 2 

......................... 50 

......................... 50 

+ 2 

+++M+MMMM+MMMM+MM+MMM+M++ 14 

X X 3 

X 1 

......................... 50 

1 

+++++++++++M+++++++++++++ 48 

......................... 50 

L 

x x 3 

X X 5 
198 Scopolamine Hydrobromide Trihydrate, TR 445 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg 

Number of Days on Study 

Carcass ID Number 

AlimentarySystem 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Anus, sarcoma 

Intestine large, cecum 

Intestine small, duodenum 

Hemangiosarcoma 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Mast cell tumor benign 

Squamous cell papilloma 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

General Body System 

None 

Genital System 

Clitoral gland 

Ovary 

Cystadenoma 

Uterus 

Hemangioma 

2333556666661111111111111 

405111014118~0222222222222 

3263084116856666888888888 

5556566555555555555555555 

4980510884118188444555666 

4574802145188169268369126 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... X 

......................... 

......................... 

......................... xx x X 

X 

X X xxx 

+ ......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... X 

M++++MM+++M+M++++++++++++ 

++++++++++M++++++++++++++ 

X X 

......................... 

++++++++M++++++++++++++++ 

......................... 

......................... A 
Lesions 199 in Female Mice 

TABLED2 

Individual Animal Tumor Pathologyof Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Anus, sarcoma 

Intestine large, cecum 

Intestine small, duodenum 

Hemangiosarcoma 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular carcinoma, multiple 

Hepatocellular adenoma 

Mesentery 

Pancreas 

Salivary glands 

Stomach, forestomach 

Mast cell tumor benign 

Squamous cell papilloma 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Adenoma 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Thyroid gland 

General Body System 

None 

Genital System 

Clitoral gland 

Ovary 

Cystadenoma 

Uterus 

Hemangioma 

7777777777777777777777777 

2222222222223333333333333 

8888888888880000000000000 

5555555556665555555555666 Total 

6678899990004446677799000 Tissues1 

7820513780581394545902169 Tumors 

......................... 50 

++++++++++++M++++++++++++ 49 

......................... 50 

......................... 50 

X 1 

......................... 50 

......................... 50 

1 

......................... 50 

++++++++++++++++++M++++++ 49 

......................... 50 

X 5 

X X 3 

X X X X 9 

+ + + 4 

......................... 50 

......................... 50 

......................... 50 

X 1 

A 1 

......................... 50 

......................... 50 

......................... 50 

......................... 50 

++++++++++++++++++++++++M 49 

......................... 50 

1 

++M++M+M++M+M++++M+M++++M 37 

++++M+++++++++++++++++++M 47 

X 3 

......................... 50 

++++++++++++++++++++++M++ 48 

++++++++++++++++++++M++++ 49 

X X X 3 

......................... 50 

1 
200 Scopolamine Hydrobromide Trihydrate, TR 445 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

HematopoieticSystem 

Bone marrow 

Lymph node 

Bronchial, carcinoma, metastatic, 

harderian gland 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

Mediastinum, rhabdomyosarcoma, 

metastatic, skeletal muscle 

IntegumentarySystem 

Mammary gland 

Skin 

Subcutaneous tissue, fibrosarcoma 

Musculoskeletal System 

Bone 

Skeletal muscle 

Rhabdomyosarcoma 

Nervous System 

Brain 

RespiratorySystem 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Carcinoma, metastatic, harderian 

gland 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Trachea 

Special Senses System 

Eye

Harderian gland 

Adenoma 

Carcinoma 

Urinary System 

Kidney 

Urinary bladder 

SystemicLesions 

Multiple organs 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

Lymphoma malignant undifferentiated 

cell type 

2333556666667777777777777 

4057170147780222222222222 

3263084776856666888888888 

5556566555555555555555555 

4980510884778788444555666 

4574802145788169268369126 

......................... + + + 

X 

++++++++++++++++++++M++++ 

++++++++++M++++++++++++++ 

......................... 

+++M+++MM++M+++++++++++++ 

X 

......................... 

.......................... 

......................... + 

X 

......................... 

......................... 

X 

X 

xxxx ......................... 

......................... 

+ + 

MMMMMMM+++M++M++M+M+M+MM+ 

X X 

X 

......................... 

......................... 

......................... X 

X xx 

X 
Lesions 201 in Female Mice 

TABLE D2 

Individual Animal Tumor Pathology of Female Mice in the 2-Y-r Gavage Study 

of ScopolamineHydrobromideTrihydrate: 5 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Hematopoietic System 

Bone marrow 

Lymph node 

Bronchial, carcinoma, metastatic. 

harderian gland 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Thymus 

Mediastinum, rhabdomyosarcoma, 

metastatic, skeletal muscle 

Integumentary System 

Mammary gland 

Skin 

Subcutaneous tissue, fibrosarcoma 

Musculoskeletal System 

Bone 

Skeletal muscle 

Rhabdomyosarcoma 

Nervous System 

Brain 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolarbronchiolar carcinoma 

Carcinoma, metastatic, harderian 

gland 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Trachea 

Special Senses System 

EY e Harderian gland 

Adenoma 

Carcinoma 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

Lymphoma malignant undifferentiated 

cell type 

7777777777777777777777777 

2222222222223333333333333 

8888888888880000000000000 

5555555556665555555555666 Total 

6678899990004446677799000 Tissues/ 

7820513780581394545902169 Tumors 

......................... 50 

+ 4 

1 

++++++++++++++++++++++++M 48 

......................... 49 

......................... 50 

......................... 46 

1 

......................... 50 

......................... 50 

X 1 

......................... 50 

1 

1 

......................... 50 

......................... 50 

X 1 

1 

1 

X 5 

......................... 50 

......................... 50 

2 

M+MMMMM+MMMMM+MMMMMM+MMMM 15 

2 

1 

......................... 50 

+++++M+++++++++++++++++++ 49 

......................... 50 

X 2 

X X X 6 

1 
202 Scopolamine Hydrobromide Trihydrate, TR 445 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Histiocytic sarcoma 

Mesentery 

Histiocytic sarcoma 

Pancreas 

Histiocytic sarcoma 

Salivary glands 

Stomach, forestomach 

Squamous cell papilloma 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Pars intermedia, adenoma 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Clitoral gland 

Ovary 

Cystadenoma 

Luteoma 

Uterus 

3344556666666777777777777 

5936450317899222222222222 

0928414607014666668888888 

6666666666666666666666666 

7466454222541135572223444 

8098651691040658952344379 

......................... 

......................... 

......................... 

++++++++++++++++++++M++++ 

......................... 

......................... 

++++++++++++M++++++++++++ 

......................... 

......................... X X 

X 

X 

X 

+ + + 

X ......................... X ......................... 

......................... X 

......................... 

......................... 

......................... 

......................... 

......................... 

......................... 

+++++++M+++++++++++++++++ 

++++++++++++++++++++M++++ 

X X X 

......................... X 

++++M++++++++++++++++++++ 

......................... 

A 

......................... 
Lesions in Female Mice 203 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

Alimentary System 

Esophagus 

Gallbladder 

Intestine large, colon 

Intestine large, rectum 

Intestine large, cecum 

Intestine small, duodenum 

Intestine small, jejunum 

Intestine small, ileum 

Liver 

Hepatocellular carcinoma 

Hepatocellular adenoma 

Hepatocellular adenoma, multiple 

Histiocytic sarcoma 

Mesentery 

Histiocytic sarcoma 

Pancreas 

Histiocytic sarcoma 

Salivary glands 

Stomach, forestomach 

Squamous cell papilloma 

Stomach, glandular 

Cardiovascular System 

Blood vessel 

Heart 

Endocrine System 

Adrenal cortex 

Adrenal medulla 

Islets, pancreatic 

Parathyroid gland 

Pituitary gland 

Pars distalis, adenoma 

Pars intermedia, adenoma 

Thyroid gland 

Follicular cell, adenoma 

General Body System 

None 

Genital System 

Clitoral gland 

Ovary 

Cystadenoma 

Luteoma 

uterus 

77777777777777777777771777 

22222223333333333333333333 

88888880000000000000000000 

66666666666666666666666666 Total 

55566781111113334566667777 Tissues/ 

26712202345781365434573469 Tumors 

++++++++++++++++++M+++++++ 50 

.......................... 51 

.......................... 51 

.......................... 50 

........................... 50 

.......................... 51 

.......................... 50 

.......................... 51 

.......................... 51 

X X 4 

X X X 4 

A 2 

1 

+ + 5 

1 

.......................... 51 

1 

.......................... 51 

.......................... 51 

X 2 

.......................... 51 

.......................... 51 

.......................... 51 

.......................... 51 

++++++++++++++++++++++++M+ 50 

.......................... 51 

+M++M+M++M+++++++++M++++++ 45 

M+++++M+++M+++++++++++M+++ 46 

3 

X 1 

.......................... 51 

X 2 

+++++++++++++++++++M++M+++ 48 

.......................... 51 

X 1 

1 

.......................... 51 
204 Scopolamine Hydrobromide Trihydrate, TR 445 

TABLED2 

Individual Animal Tumor Pathology of Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

HematopoieticSystem 

Bone marrow 

Hemangiosarcoma 

Lymph node 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Histiocytic sarcoma 

Thymus 

Mast cell tumor malignant 

IntegumentarySystem 

Mammary gland 

Skin 

Squamous cell carcinoma 

Musculoskeletal System 

Bone 

Nervous System 

Brain 

Peripheral nerve 

Spinal cord 

Respiratory System 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Trachea 

Special Senses System 

Harderian gland 

Adenoma 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

3344556666666111111111111 

593645031189922.2222222222 

0928414607014666668888888 

6666666666.666666666666666 

7466454222541135512223444 

8098651691040658952344379 

......................... 

+ ++ 

X 

......................... 

++++++++++M++++M+++++++++ 

......................... A 

+++++++MM+M++++++++++++M+ 

X 

......................... 

......................... X 

......................... 

......................... 

M 

+ 

......................... 

X 

X 

......................... 

......................... 

M+MMM+MMMM++MMM+M+M+’MMMMM 

......................... 

......................... 

.......................... X 

x x x X 
--- 

Lesions in Female Mice 205 

TABLE D2 

Individual Animal Tumor Pathologyof Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate: 25 mg/kg (continued) 

Number of Days on Study 

Carcass ID Number 

HematopoieticSystem 

Bone marrow 

Hemangiosarcoma 

Lymph node 

Mediastinal, histiocytic sarcoma 

Lymph node, mandibular 

Lymph node, mesenteric 

Spleen 

Histiocytic sarcoma 

Thymus 

Mast cell tumor malignant 

Integumentary System 

Mammary gland 

Skin 

Squamous cell carcinoma 

Musculoskeletal System 

Bone 

Nervous System

Brain 

Peripheral nerve 

Spinal cord ~ ~ 

Respiratory System ____ 

Lung 

Alveolar/bronchiolar adenoma 

Alveolar/bronchiolar carcinoma 

Hepatocellular carcinoma, metastatic, 

liver 

Nose 

Trachea 

Special Senses System 

Harderian gland 

Adenoma 

Urinary System 

Kidney 

Urinary bladder 

Systemic Lesions 

Multiple organs 

Histiocytic sarcoma 

Lymphoma malignant lymphocytic 

Lymphoma malignant mixed 

77777777777777777777777777 

22222223333333333333333333 

88888880000000000000000000 

66666666666666666666666666 Total 

55566781111113334566667777 Tissues/ 

26712202345781365434573469 Tumors 

.......................... 51 

X 1 

3 

1 

.......................... 51 

+++++++++++++++++++M++++++ 48 

.......................... 51 

1 

++++++++++M++++++++M++++++ 45 

1 

.......................... 51 

.......................... 51 

1 

.......................... 51 

.......................... 51 

1 

- ~- ~~ 

.......................... 51 

X X 2 

1 

1 

.......................... 51 

.......................... 51 

M++M+MM+M+MMMM+MMMMMMMMMMM 13 

X 1 

.......................... 51 

.......................... 51 

.......................... 51 

1 

4 

xx X 3 
206 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLED3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate 

Vehicle Control 1 mgk 

HarderianGland:Adenoma 

OvedI ratea 1/51 (2%) 3/50 (6%) 2/50 (4%) 1/51 (2%) 

Adjusted rateb 3.0% 7.5% 5.1% 2.6% 

Terminal rate' 1/33 (3%) 2/36 (6%) 1/37 (3%) 1/38 (3%) 

First incidence (days) 726 (T) 482 676 726 (T) 

Life table testd P=0.380N P=O.328 P=O.532 P=0.730N 

Logistic regression testd P=0.402N P=O.285 P=O.497 P=0.730N 

Cochran-Armita e testd P=0.402N 9 Fisher exact test P=O.301 P=O.492 P=0.752N 

HarderianGland:Adenoma or Carcinoma 

Overall rate 3/51 (6%) 4/50 (8%) 3/50 (6%) 1/51 (2%) 

Adjusted rate 7.2% 10.2% 7.5% 2.6% 

Terminal rate 1/33 (3%) 3/36 (8%) 1/37 (3%) 1/38 (3%) 

First incidence (days) 511 482 676 726 (T) 

Life table test P=0.157N P=O.S22 P=0.642N P=0.285N 

Logistic regression test P=0.169N P=O.450 P=O.653 P=0.320N 

Cochran-Armitage test P=0.166N 

Fisher exact test P=O.489 P=O.652 P=0.309N 

Liver:HepatocellularAdenoma 

Overall rate 15/51 (29%) 18/50 (36%) 9/50 (18%) 6/51 (12%) 

Adjusted rate 42.3% 47.1% 22.8% 15.4% 

Terminal rate 13/33 (39%) 16/36 (44%) 7/37 (19%) 5/38 (13%) 

First incidence (days) 604 676 578 694 

Life table test P=0.004N P=O.439 P=0.077N P=O.OllN 

Logistic regression test P=0.005N P=O.368 P=0.119N P=0.017N 

Cochran-Armitage test P=0.007N 

Fisher exact test P=O.311 P=0.133N P=0.024N 

Liver:HepatocellularCarcinoma 

Overall rate 8/51 (16%) 6/50 (12%) 8/50 (16%) 4/51 (8%) 

Adjusted rate 21.1% 16.0% 19.1% 10.3% 

Terminal rate 5/33 (15%) 5/36 (14%) 4/37 (1 1 %) 3/38 (8%) 

First incidence (days) 594 680 617 694 

Life table test P=0.145N P=0.337N P=0.542N P=0.139N 

Logistic regression test P=0.151N P=0.381N P=O.571 P=0.170N 

Cochran-Armitage test P=0.167N 

Fisher exact test P=0.403N P=O.590 P=0.179N 

Liver:HepatocellularAdenoma or Carcinoma 

Overall rate 2215 1 (43 %) 21/50 (42%) 16/50 (32%) 9/51 (18%) 

Adjusted rate 57.1% 55.0% 37.0% 23.1% 

Terminal rate 17/33 (52%) 19/36 (53 %) 10137 (27%) 8/38 (21 %) 

First incidence (days) 594 676 578 694 

Life table test P=O.OOlN P=0.371N P=0.106N P=0.002N 

Logistic regression test P=0.002N P=0.466N P=0.165N P=0.003N 

Cochran-Armitage test P=0.002N 

Fisher exact test P=0.534N P=0.171N P=0.005N 
Lesions in Female Mice 207 

TABLED3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle Control 

Lung:Alveolar/bronchiolarAdenoma 

Overall rate 3/51 (6%) 

Adjusted rate 8.0% 

Terminal rate 1/33 (3%) 

First incidence (days) 618 

Life table test P=0.354N 

Logistic regression test P=0.381N 

Cochran-Armitage test P=0,387N 

Fisher exact test 

Lung:AlveolarlbronchiolarCarcinoma 

Overall. rate 1/51 (2%) 

Adjusted rate 3.0% 

Terminal rate 1/33 (3%) 

First incidence (days) 726 (T) 

Life table test P=0.431N 

Logistic regression test P=0.440N 

Cochran-Armitage test P=0.439N 

Fisher exact test 

Lung:AlveolarlbronchiolarAdenoma or Carcinoma 

Overall rate 4/51 (8%) 

Adjusted rate 10.8% 

Terminal rate 2/33 (6%) 

First incidence (days) 618 

Life table test P-0.256N 

Logistic regression test P=0.277N 

Cochran-Armitage test P=0.280N 

Fisher exact test 

Ovary:Cystadenoma 

Overall rate 3/51 (6%) 

Adjusted rate 7.1 % 

Terminal rate 0133 (0%) 

First incidence (days) 511 

Life table test P=0.406N 

Logistic regression test P=0.426N 

Cochran-Armitage test P=0.427N 

Fisher exact test 

PituitaryGland(ParsDistalis):Adenoma 

Overall rate 4/50 (8%) 

Adjusted rate 11.8% 

Terminal rate 3/33 (9%) 

First incidence (days) 720 

Life table test P=0.525N 

Logistic regression test P=0.562N 

Cochran-Armitage test P=0.557N 

Fisher exact test 

1 mg/kg 

4/50 (8%) 

11.1% 

4/36 ( 1 1%) 

726 (T) 

P=O.535 

P=O.506 

P=O.489 

3150 (6%) 

7.3% 

1136 (3 %) 

590 

P=O.326 

P=O.298 

P=O.301 

7/50 (14%) 

17.9% 

5/36 (14%) 

590 

P=O.302 

P=O.262 

P=O.251 

0/49 (0%) 

0.0% 

0135 (0%) -e 

P=0.124N 

P=0.142N 

P=0.129N 

3/47 (6%) 

8.8% 

3/34 (9%) 

726 (T) 

P=0.482N 

P=0.505N 

P=0.535N 

5 mg/kg 

1/50 (2 %) 

2.7% 

1/37 (3%) 

726 (T) 

P=0.286N 

P=0.312N 

P=0.316N 

1/50 (2%) 

2.6% 

0137 (0%) 

706 

P=0.748N 

P=0.762N 

P=O.748 

2/50 (4%) 

5.3% 

1/37 (3 %) 

706 

P=0.313N 

P=0.344N 

P=0.348N 

3/49 (6%) 

8.3% 

3/36 (8%) 

726 (T) 

P=0.643N 

P=O.645 

P=O.642 

3/47 (6%) 

7.5% 

1/35 (3%) 

604 

P=0.475N 

P=0.537N 

P-0.535N 

25 mg/kg 

2/51 (4%) 

5.3% 

2/38 (5 X) 

726 (T) 

P=0.453N 

P=0.493N 

P=0.500N 

1/51 (2%) 

2.2% 

0138 (0%) 

604 

P=0.745N 

P=O.760 

P=0.752N 

3/51 (6%) 

7.4% 

2/38 (5%) 

604 

P=0.450N 

P=0.496N 

P=0.500N 

1/51 (2%) 

2.6% 

1/38 (3%) 

726 (T) 

P=0.289N 

P=0.316N 

P=0.309N 

3/46 (7%) 

8.2% 

2/33 (6%) 

670 

P=0.484N 

P=0.543N 

P=0.547N 
208 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLED3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

ThyroidGland(FollicularCell):Adenoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

AllOrgans:Hemangiosarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

All Organs:Hemangioma or Hemangibsarcoma 

Overall rate 

Adjusted rate 

Terminal rate 

First incidence (days) 

Life table test 

Logistic regression test 

Cochran-Armitage test 

Fisher exact test 

VehicleControl 1 mgkg 5 mgkg 25 mgkg 

3/51 (6%) 0150 (0%) 0150 (0%) 2/51 (4%) 

9.1% 0.0% 0.0% 4.9% 

P=O.509 

726 (9 

3/33 (9%) 0136 (0%) 

P=0.106N 

- 0137 (0%) 

P=O.lOlN 

- P=0.442N 

677 

1/38 (3%) 

P=O.488 P=0.106N P=O.lOlN P=0.483N 

P=O.482 

P=0.125N P=O.lZSN P=0.500N 

4/51 (8%) 2/50 (4%) 1/50 (2%) 1/51 (2%) 

11.3% 4.5 % 2.7% 2.6% 

P=0.222N 

709 

2/33 (6%) 

P=0.322N 

562 

0136 (0%) 

P=0.159N 

726 (9 

1/37 (3%) 

P=0.151N 

726 (T) 

1/38 (3%) 

P=0.234N P=0.344N P=O.l7ON P=0.162N 

P=0.235N 

P=0.348N P=0.187N P=0.181N 

4/51 (8%) 2/50 (4%) 2/50 (4%) 1/51 (2%) 

11.3% 4.5% 5.1% 2.6% 

2/33 (6%) 0136 (0%) 1/37 (3%) 1/38 (3%) 

P=0.207N 

709 

P-0.322N 

562 

P=0.307N 

678 

P-0.151N 

726 (T) 

P=0.217N P=0.344N P=0.336N P=0.162N 

P=0.219N 

P=0.348N P=0.348N P=0.181N 

AllOrgans:MalignantLymphoma(Lymphocytic,Mixed, or UndifferentiatedCellType) 

Overall rate 9/51 (18%) 8/50 (16%) 9/50 (18%) 7/51 (14%) 

Adjusted rate 23.0% 21.6% 22.6% 16.2% 

Terminal rate 5/33 (15%) 7/36 (19%) 7/37 (19%) 3/38 (8%) 

First incidence (days) 708 581 510 55 1 

Life table test P=0.312N P=0.447N P=0.531N P=0.332N 

Logistic regression test P=0.355N P=0.497N P=O.585 P=0.393N 

Cochran-Armitage test P=0.359N 

Fisher exact test P=0.518N PtO.584 P=0.393N 

AllOrgans:Benign Neoplasm 

Overall rate 24/5 1 (47 46) 25/50 (50%) 21/50 (42%) 18/51 (35%) 

Terminal rate 

Adjusted rate 

18/33 (55%) 

60.9% 

21/36 (58%) 

62.0% 

15/37 (41 4%) 

48.7% 

15/38 (39%) 

43.8% 

First incidence (days) 482 511 578 670 

Life table test P-0.057N P=0.518N P=0.235N P=0.072N 

Cochran-Armitage test 

Logistic regression test 

P=0.090N 

P=O.512 P=0.079N P=0.373N P=0.129N 

Fisher exact test P=O.462 P=0.378N P=0.157N 
Lesions in Female Mice 209 

TABLED3 

Statistical Analysis of Primary Neoplasms in Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

All Organs:MalignantNeoplasms 

Overall rate 2615 1 (51%) 22/50 (44%) 2 1/50 (42I) 14/51 (27%) 

Adjusted rate 54.8% 49.5% 47.7% 31.6% 

Terminal rate 12/33 (36%) 14/36 (39%) 14/37 (38%) 8/38 (21X) 

First incidence (days) 492 532 510 55 1 

Life table test P=0.017N P=0.247N P-0.185N P=0.016N 

Logistic regression test P=0.017N P=0.389N P=0.425N P-0.025N 

Cochran-Armitage test P=0.012N 

Fisher exact test P=0.308N P=0.240N P=0.013N 

All Organs:Benign or MalignantNeoplasms 

Overall rate 40/51 (78X) 36/50 (72%) 35/50 (70%) 29/5 1 (57 X) 

Adjusted rate 83.2% 78.0% 76.1% 64.3 % 

Terminal rate 25/33 (76%) 26/36 (72%) 26/37 (70%) 22/38 (58%) 

First incidence (days) 492 482 510 55 1 

Life table test P=0.019N P=0.196N P=0.131N P=0.013N 

Logistic regression test P=0.021N P=0.322N P=0.229N P=0.029N 

Cochran-Armitage test P=0.014N 

Fisher exact test P=0.302N P=O.229N P=0.017N 

(T)Tenninal sacrifice 

a Number of neoplasm-bearing animalslnumber of animals examined. Denominator is number of animals examined microscopically for liver, lung, pituitary 

gland, and thyroid gland; for other tissues, denominator is number of animals necropsied. 

Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality 

Observed incidence at terminal kill * Beneath the control incidence are the P values associated with the trend test. Beneath the dosed group incidence are the P values corresponding to 

painvise 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 ail tests, a negative trend or a lower incidence in a dose group is indicated by N. e Not applicable; no neoplasmsinanimalgroup 
210 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLED4 

Historical Incidence of Hepatocellular Neoplasms in Female B6C3Fl Mice Administered Waterby Gavagea 

Incidence in Controls 

Adenoma CarcinomaAdenomaor Carcinoma 

Overall Historical Incidence 

Total 131315 (4.1%) 8/315 (2.5%) 211315 (6.7%) 

Standard deviation 3.2% 2.1% 4.2% 

Range 2%-10% 0%-6% 2%-12% 

a Data as of 17 June 1994 
Lesions in Female Mice 211 

TABLED5 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydratea 

VehicleControl 

DispositionSummary 

Animals initially in study 70 70 70 

IS-Month interim evaluationb 19 20 20 

Early deaths 

Accidental deaths 2 1 

Moribund 9 11 7 

Natural deaths 7 3 5 

Survivors 

Terminal sacrifice 33 36 37 

Animals examined microscopically 61 60 60 

15-Month Interim Evaluation 

Alimentary System 

Esophagus (10) 

Periesophageal tissue, inflammation, 

suppurative 

Liver (10) 

Basophilic focus 

Inflammation 

Mixed cell focus 1 (10%) 

Necrosis 

Pancreas (10) 

Atrophy 1(10%) 

Stomach,forestomach (10) 

Hyperplasia, focal 2 (20%) 

Cardiovascular System 

Blood vessd (10) 

Aorta, inflammation, chronic active 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

Parathyroid gland 

cyst 

Inflammation, chronic active 

Pituitary gland 

Pars distalis, hyperplasia 

Thyroid gland 

Follicular cell, hyperplasia 

a Number of animals examined microscopically at the site and the number of animals with lesion 

Nine or ten animals per group received ophthalmic examinations and were discarded without further evaluation 

25 mgkg 

70 

19 

7 

6 

38 

61 
212 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLED5 

Summary of the Incidence of Nonneoplastic Lesionsin Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

15-Month Interim Evaluation (continued) 

GenitalSystem 

Clitoral gland 

Duct, ectasia 

(10) (10) 

1 (10%) 

(10) 

Ovary 

cyst 

(10) 

1 (10%) 

(10) 

3 (30%) 

(10) 

1 (10%) 

uterus 

Hyperplasia, cystic 

(10) 

5 (50%) 

(10) 

8 (80%) 

(10) 

4 (40%) 

HematopoieticSystem 

Bone mahw 

Myelofibrosis 

Thymus 

Inflammation, chronic active 

Mineralization 1 (11%) 

IntegumentarySystem 

Skin 

Inflammation, chronic active 

Respiratory System 

Lung (10) (10) 

Inflammation, chronic active 1 (10%) 

Alveolar epithelium, hyperplasia 1 (10%) 

Special Senses System 

Harderian gland 

Inflammation, chronic active 

Urinary System 

Kidney (10) (10) (10)

Nephropathy 5 (50%) 3 (30%) 1 (10%) 

Artery, inflammation, chronic active 1 (10%) 

Systems Examined With No Lesions Observed 

General Body System 

MusculoskeletalSystem 

NervousSystem 

(10) 

(10) 

2 (20%) 

(10) 

4 (40%) 

(10) 

2 (20%) 
Lesions in Female Mice 213 

TABLEDS 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide "hydrate (continued) 

2-Year Study 

Alimentary System 

Esophagus 

Perforation 

Periesophageal tissue, inflammation, 

suppurative 

Intestine large, rectum 

Inflammation, chronic active 

Intestine small, duodenum 

Ulcer 

Intestine small, jejunum 

Hyperplasia, lymphoid 

Inflammation, chronic active 

Ulcer 

Liver 

Angiectasis 

Basophilic focus 

Clear cell focus 

Developmental malformation 

Eosinophilic focus 

Fatty change 

Hematopoietic cell proliferation 

Inflammation, chronic active 

Mixed cell focus 

Necrosis 

Kupffer cell, pigmentation, hemosiderin 

Mesentery 

Artery, inflammation, chronic active 

Fat, necrosis 

Pancreas 

Atrophy 

HYPe~PhY 

Inflammation, chronic active 

Duct, cyst 

Salivary glands 

Artery, inflammation, chronic active 

Duct, hyperplasia 

Stomach,forestomach 

Diverticulum 

Hyperplasia, focal 

Infiltration cellular, plasma cell 

Stomach, glandular 

Erosion 

Cardiovascular System 

Blood vessel (51) (50)

Aorta, inflammation, chronic active 1 (2%)

Heart (51) (50)

Inflammation, chronic active 1 (2%) 

Mineralization 2(4%)

Artery, inflammation, chronic active 2 (4%) 
214 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLED5 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

. VehicleControl 1 mgkg 5 mgkg 25 mgkg 

2-YeUr study (continued) 

Endocrine System 

Adrenal cortex 

Accessory adrenal cortical nodule 

Hematopoietic cell proliferation 

Hyperplasia 

Capsule, hyperplasia, adenomatous 

Adrenal medulla 

Hyperplasia 

Islets, pancreatic 

Hyperplasia 

Pituitary gland 

Angiectasis 

Pars distalis, hyperplasia 

Pars intermedia, hyperplasia 

Thyroid gland 

Inflammation, chronic active 

Follicular cell, hyperplasia 

General Body System 

None 

Genital System 

Clitoral gland 

Duct, ectasia 

Ovary 

cyst 

Inflammation, suppurative 

Mineralization 

Uterus 

Angiectasis 

Hyperplasia, cystic 

Inflammation, chronic active 

HematopoieticSystem 

Bone marrow 

Myelofibrosis 

Erythroid cell, hyperplasia 

Myeloid cell, hyperplasia 

Lymph node 

(50) (51) 

Mediastinal, ,hyperplasia, lymphoid 

Renal, angiectasis 

Lymph node, mandibular 

Hematopoietic cell proliferation 

Hyperplasia, lymphoid 

Lymph node, mesenteric 

Hyperplasia, lymphoid 

. . 

(5 1) 

1 (2%) 

1 (2%) 1 (2%) 1 (2%) 

1 (2%) 

(49) (49) 

2 (4%) 

(50) 

(47) 

1 (2%) 

24 (48%) 11 (23%) 13 (28%) 

1 (2%) 

(50) (50) (5 1) 

5 (10%) 1 (2%) 

14 (28%) 10 (20%) 8 (16%) 

(5 1) (49) 

16 (31%) 14 (29%) 

1 (2%) 

1 (2%) 

1 (2%) 

38 (75%) 29 (58%) 

1 (2%) 1 (2%) 

(51) (5 1) 

22 (43%) 13 (25%) 

3 (6%) 

9 (18%) 3 (6%) 

(6) (3) 

1 (20%) 

(46) 

1 (2%) 

1 (2%) 1 (2%) 1 (2%) 

(50) (47) (49) (48) 

1 (2%) 1 (2%) 2 (4%) 
Lesions in Female Mice 215 

TABLED5 

Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

&Year Study (continued) 

Hematopoietic System (continued) 

Spleen 

Depletion lymphoid 

Hematopoietic cell proliferation 

Hyperplasia, lymphoid 

Hyperplasia, plasma cell 

Thymus 

Atrophy 

IntegumentarySystem 

Mammary gland 

Hyperplasia 

Skin 

Cyst epithelial inclusion 

Inflammation, chronic active 

Ulcer 

Subcutaneous tissue, fibrosis, chronic active 

MusculoskeletalSystem 

None 

NervousSystem 

Brain 

Infarct 

Artery, inflammation, chronic active 

Neuron, necrosis 

Peripheral nerve 

Degeneration 

Spinal cord 

White matter, degeneration 

RespiratorySystem 

Lung 

Inflammation, chronic active 

Alveolar epithelium, hyperplasia 

(51) 

2 (4%) 

17 (33%) 

3 (6%) 

1 (2%) 

(48) 

3 (6%) 

(51) 

1 (2%) 

(51) 

1 (2%) 

1 (2%) 

(51) 

1 (2%) 

(50) 

11 (22%) 

(43) 

7 (16%) 

(49) 

3 (6%) 

(50) 

1 (2%) 

(50) 

2 (4%) 

1 (2%) 

(50) 

17 (34%) 

2 (4%) 

(46) 

4 (9%) 

(50) 

2 (4%) 

(50) 

(50) 

(51) 

7 (14%) 

2 (4%) 

(45) 

3 (7%) 

(51) 

4 (8%) 

(51) 

1 (2%) 

(51) 

Trachea (50) (51) (50) (51) 

Artery, inflammation, chronic active 1 (2%) 

E Ye 

Special Senses System 

Retina. atrophy 

Lens, cataract 

Cornea, inflammation, chronic active 

Degeneration 

(2) 

2 (100%) 

(2) 

1 (50%) 

(2) 

1 (50%) 

1 (50%) 

1 (50%) 
216 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLED5 

Summary of the Incidence of Nonneoplastic Lesionsin Female Mice in the 2-YearGavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle Control 1 mI3h 

Kidney 

Urinary System 

&Year study (continued) 

Cyst 

Infarct 

(5 1) 

1(2%) 

1 (2%) 

(50) (50) (51) 

Inflammation, chronic active 1(2%) 

Nephropathy 23 (45%) 21 (42%) 26 (52%) 10 (20%) 

Renal tubule, necrosis, acute 

Artery, inflammation, chronic active 

Inflammation, chronic active 

Urinary bladder 

1 (2%) 

(50) (48) 

1(2%) 

(49) 

1(2%) 

(51) 

1 (2%) 

Artery, inflammation, chronic active 2(4%) 1 (2%) 
217 

APPENDIX E 

GENETIC TOXICOLOGY 

S~OEELAMUTAGENICITYTEST PROTOCOL . .. . .. . . .. . . . . . . . . . . . . . . .. . .... . . . . 218 

CHINESEHAM!~TER OVARYCELL CYTOGENETICS PROTOCOLS.. . .. . .. . . . . ... . . . . . . . . . . 218 

MOUSEPERIPHERAL BLOODMICRONUCLEUS TEST PROTOCOL .. . .. . .. . ... . .. . . . .. . .. . . 219

RESULTS ......................................................... 220 

TABLEEl Mutagenicity of Scopolamine Hydrobromide Trihydrate in Salmonella typhimurium . . . 221 

TABLEE2 Induction of Sister Chromatid Exchanges in Chinese Hamster Ovary Cells 

by Scopolamine Hydrobromide Trihydrate . . . . . . . . . .. . . . . . . . . . . . . .... .. 223 

TABLEE3 Induction of Chromosomal Aberrations in Chinese Hamster Ovary Cells 

by Scopolamine Hydrobromide Trihydrate ... . . . .. . .. . . . . . . . .. . . . . . . . . . 225 

TABLEE4 Frequency of Micronuclei in Mouse Peripheral Blood Erythrocytes 

Following Treatment with Scopolamine Hydrobromide Trihydrate by Gavage 

for14Weeks ................................................ 226 
218 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

GENETIC TOXICOLOGY 

SALMONELLAMUTAGENICITYTESTPROTOCOL 

Testing was performed as reported by Zeiger et al. (1988). Scopolamine hydrobromide trihydrate was 

sent to the laboratory as a coded aliquot from Radian Corporation (Austin, TX). It was incubated with the 

Salmonella typhimrium tester strains TA97, TA98, TA100, TA1535, and 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 37" C. Top agar supplemented with l-histidine and d-biotin was 

added, and the contents of the tubes were mixed and poured onto the surfaces of minimal glucose agar 

plates. Histidine-independent mutant colonies arising on these plates were counted following incubation 

for 2 days at 37" C. 

Each trial consisted of triplicate plates of concurrent positive and negative controls and at least five doses 

of scopolamine hydrobromide trihydrate. The selected high dose was 10,000 pg/plate. All negative trials 

were repeated. 

In this assay, a positive response is defined as a reproducible, dose-related increase in histidineindependent (revertant) colonies in any one straixdactivation combination. An equivocal response is 

defined as an increase in revertants that is not dose related, is not reproducible, or is of insufficient 

magnitude to support a determination of mutagenicity. A negative response is obtained when no increase 

in revertant colonies is observed following chemical treatment. There was no minimum percentage or fold 

increase required for a chemical to be judged positive or weakly positive. 

CHINESEHAMSTER OVARYCELL CYTOGENETICS PROTOCOLS 

Testing was performed as reported by Galloway et al. (1987). Scopolamine hydrobromide trihydrate 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 

scopolamine hydrobromide trihydrate; the high dose was limited by toxicity in the trials conducted without 

S9; with S9, no toxicity was noted and 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 Chroni& Exchange Test: In the SCE test without S9, CHO cells were incubated for 26 hours 

with scopolamine hydrobromide trihydrate in supplemented McCoy's 5A medium. Bromodeoxyuridine 

(BrdU) was added 2 hours after culture initiation. After 26 hours, the medium containing scopolamine 

hydrobromide trihydrate 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 scopolamine 

hydrobromide trihydrate, serum-free medium, and S9 for 2 hours. The medium was then removed and 

replaced with medium containing serum and BrdU and no scopolamine hydrobromide trihydrate and 

incubation proceeded for an additional 26 hours, with Colcemid present for the final 2 hours. Harvesting 

and staining were the same as for cells treated without S9. All slides were scored blind and those from a 

single test were read by the same person. Fifty second-division metaphase cells were scored for frequency 

of SCEdcell from each dose level. Because a shift in the pH was noted at doses of 2,000 pg/mL and 

above in the trials conducted with S9, N-(2-hydroxyethyl)- piperazine-N'-(2-ethanesulfonic acid) (HEPES) 

buffer was added to the culture medium to maintain optimum pH in the third trial with S9. 
Genetic Toxicology 219 

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 <0.05) 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 scopolamine hydrobromide trihydrate for 10.5 hours; Colcemid was added and incubation continued 

for 2 hours. The cells were then harvested by mitotic shake-off, fixed, and stained with Giemsa. For the 

Abs test with S9, cells were treated with scopolamine hydrobromide trihydrate and S9 for 2 hours, after 

which the treatment medium was removed and the cells were incubated for 1 1 to 1 1.5 hours in fresh 

medium, with Colcemid present for the final 2 hours. Cells were harvested in the same manner as for the 

treatment without S9. As with the SCE test, an alteration in pH was observed in the first trial conducted 

with S9 and the second trial with S9 was conducted with HEPES buffer present in the medium to stabilize 

pH* 

Cells were selected for scoring on the basis of good morphology and completeness of karyotype 

(21 f 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). 

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 (PSO.05) difference for one dose point and a 

significant trend (P IO.015) were considered weak evidence for a positive response; significant differences 

for two or more doses indicated the trial was positive. A positive tre.nd 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. 

MOUSEPERIPHERAL BLOOD MICRONUCLEUS TETPROTOCOL 

A detailed discussion of this assay can be found in MacGregor et al. (1990). Peripheral blood samples 

were obtained from male and female B6C3F, mice at the end of the 14-week toxicity study. Smears were 

immediately prepared, and fixed in absolute methanol, stained with a chromatin-specific fluorescent dye 

mixture of Hoechst 3325Npyronin Y (MacGregor et al., 1983), and coded. Slides were scanned at 630 or 

lOOOX magnification with a semi-automated image analysis system to determine the frequency of 

micronuclei in 10,000 normochromatic erythrocytes (NCEs) in.each of 10 animals per dose group. The 

criteria of Schmid (1976) were used to define micronuclei, with the additional requirement that the 

micronuclei exhibit the characteristic fluorescent emissions of DNA (blue with 360 nm and orange with 

540 nm UV illumination); the minimum size limit was approximately one-twentieth the diameter of the 

NCE cell. 

The results were tabulated as the mean of the pooled results from all animals within a treatment group, 

plus or minus the standard error of the mean. The frequency of micronucleated cells among NCEs was 

analyzed by a statistical software package that tested for increasing trend over exposure groups with a 
220 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

one-tailed Cochran-Armitage trend test, followed by pairwise comparisons between each exposure group 

and the control group (Margolin et al., 1990). In the presence of excess binomial variation, as detected 

by a binomial dispersion test, the binomial variance of the Cochran-Armitage test was adjusted upward in 

proportion to the excess variation. In the micronucleus test an individual trial was considered positive if 

the trend test P value was less than or equal to 0.025 or the P value for any single exposure group was 

less than or equal to 0.025 divided by the number of exposure groups. A final call of positive for 

micronucleus induction is preferably based on reproducible positive trials (as noted above). Ultimately, 

the final call was determined by the scientific staff after considering the results of statistical analyses, 

reproducibility of any effects observed, and the magnitudes of those effects. 

RESULTS 

Scopolamine hydrobromide trihydrate (100 to 10,000 pg/plate) did not induce mutations in Salmonella 

typhimurium strains TA97, TA98, TA100, TA1535, or TA1537, with or without Aroclor 1254-induced 

male Sprague-Dawley rat or Syrian hamster liver S9 (Table El). In cytogenetic tests with cultured CHO 

cells, no convincing induction of SCEs was noted with scopolamine hydrobromide trihydrate doses up to 

500 pglmL without S9 or 5,000 pg/mL with S9 (Table E2). Results from the first two trials conducted 

with S9, which appeared to clearly demonstrate a significant increase in SCEs, were called into question 

by the observation of a pH shift in the culture medium produced by the high doses (2,000 pg/mL and 

higher) of scopolamine hydrobromide trihydrate which coincided with the increases in SCE. Therefore, a 

third trial was conducted, in which the pH of the culture medium was adjusted with HEPES buffer. 

Results of this third trial were negative and the overall assay .results were also considered to be negative. 

The increases in SCEs noted in the presence of S9 were attributed to the alteration in pH produced by 

high concentrations of scopolamine hydrobromide trihydrate. No induction of Abswas observed in 

cultured CHO cells treated with scopolamine hydrobromide trihydrate without S9, but with S9, even in the 

presence of HEPES buffer to maintain optimum pH, increases in the percentage of cells with Abs were 

noted in each of two trials at the highest dose tested(5,000 pg/mL) (Table E3). 

Despite the evidence for induction chromosomal damage in cultured CHO cellsin vitro, no increase in the 

frequency of micronucleated NCEs was noted in peripheral blood samples obtained from male and female 

mice at the termination of the 14-week toxicity studies with scopolamine hydrobromide trihydrate 

(Table E4). 
Gendle Toxicology 

TABLE El 

Mutagenicity or Scopolamine Hydrobromide Trthydrate in Salmonella typhimuriurr 

b Revertanu/plate

Strain Do>' -SO +hamster S9 +rat 59 

(,ta:!plate) Trial I Trial 2 10'10 30'10 '0'10 30'10 

TAlOO 0 102 t 6.4 102 ± 7.8 17 ± 8.1 101 ± U 101 ± '.0 108 t 8.4 

100 117 ± 11.$ 9S ± 8.8 90 t 2.6 106 ± S.8 90± 3.2 106 ± 4.7 

333 100 ± 4.6 94 ± 2.3 101 ± 7.S lOS ± 4.6 91± 3.• lOS ± 3.3 

1.000 87 t 6.1 108 ± 2.4 90 ± 7.4 107 ± 7.2 88' 86 t 2.7 

3,333 90 ± 10.0 96 ± 4.3 98 ± 7.3 122±·6.7 94 ± 8.S 113 ± 4.4 

10,000 98 ± 34.0 94 ± 8.0 8S ± S.I 97 ± 2.3 83 ± 2.6 107 ± 4.7 

Trial summary Neaative Nelative Negative Nelative Negative Negative d Positive control 281 ± 11.8 S69 ± 21.2 283 ± 11.6 426 ± 4.7 309 ± 1.5 294 ± 9.1 

TAlS)S 0 21 ± 1.3 2,. 0.3 1I± 0.7 17± 2.1 '± 3.S IS ± 2.3 

100 19 ± 2.6 3S ± 4.2 13± 3.S l2± 2.0 '± 1.2 13 ± l.S

333 21 ± U 33 ± 2.3 l2± 2.3 II ± 0.9 1I± 0.' l2± I., 

1.000 21 ± S.2 3O± 1.5 IO± 1.7 8 ± 0.7 IO± 0.' l2± 1.5

3,333 28 ± 1.5

10,000 I.' 29 ± 6.0 13± U 13 ± 0.7 2.2 12± 

23± 36 ± 6.0 0.7 9± 1.5 I" l2± 1.3 12 ± 0.3 I.'

Trial summary Ncgative Negative Ncgativc 

.. 

Negative Ncgativc Ncgative 

Positive conlrol 307 ± 8.2 369 ± 6.7 68 ± 4.S 96 ± 12.4 202 ± 9.S 69 ± O.S 

TAl!)? 

0 

.. 

6 ± 1.2 IO± 2.7 2l± 2.3 1I± 1.5 2l± 2.8 8± 1.0 

100 8 ± 2.0 20 ± 4.2 18± 1.7 '± 1.7 IO± U S± 2.2

333 S ± 0.3 18 ± 0.9 23± 2.0 I.' 1I± 2.1 2S ± 1.5 

1.000 1.8 21 ± 2.7 2O± 

... 0.7 

10 ± 0.7 20 ± 2.0 8± 1.5 

3,333 7± 1.0 20 ± 2.9 2O± 2.' 10 ± 1.2 21 ± 1.2 

.. 

1± 1.7 c 10,000 6 ± 2.7 12 ± 1.5 2l± 8 ± 2.2 2S ± 2.2 S± I.S 

Trial summary NClative Negative Nelative Ncgative NClative NC8ative 

Positive control 82S ± 65.6 38 ± S.2 46S ± IS.7 72 ± 2.9 60 ± 11.1 28± I.S 

TA9S 0 I" 3.0 21 ± 2.8 37± 1.5 31± 2.2 "± 3.8 23± 1.7 

100 11± 1.0 17 ± 3.7 33± 1.2 33± 3.S 33± 1.3 23± 2.' 

333 18± 1.7 14 ± 0.9 32± 3.1 28 ± S.2 31 ± 4.4 ,,± '.S 

1.000 16 ± 6.0 12 ± U 27 ± 0.9 30 ± S.6 30 ± 3.8 26± 3.S 

3,333 1$ ± 10.0 II ± 1.9 30 ± 2.$ 30 ± 2.3 33 ± 2.S 29± 1.7 

10,000 14 ± 3.S 20 ± 3.6 27± 1.0 28 ± 3.6 2S ± S.2 2S ± 2.6 

Trial summary 

Positive CODtroI 

Nc,ltive 

114 ± S.2 

Nelative 

224 ± 13.6 

Neptivc 

8S ± 3.2 

NClative 

7. ± 9.7 

Nc,uive 

70 ± 4.7 

Nc,ative 

109 ± 9.0 
222 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLE El
 

Mutagenicity or Scopolamine Hydrobromide Trihydrate in Salnwntlla typhimurium (continued)
 

Revertants/plate 

Strain IloR 

(pglpbtr) 

-8' 

Trial. Trial 2 

+hamster 59 I." 3... ,... +rat 89 

3... 3... 

TA97 79. 2.6 72, '.8 106 ± 5.6 200 ± 16.6 122 ± 11.7 182 ± 1.7 133 ± 13.0 • 14. 7.1 118 ± 8.) 

m 14. 3.4 7" 8.1 117 ± 8.1 172 ± 10.7 123 ± 5.2 201 ± 6.0 178 ± 6.1 

1,000 79. ,.• 71. U liS ± 6.6 136 ± 12.0 114 ± 7.8 185 ± 8.0 164±It.3 

100 79. 3.• 211 ± 22.3 120 ± 7.4 197 ± 20.1 168 ± 7.6 

3.333 83. 3.3 59. 1.2 106 ± 4.0 286 ± 21.1 118 ± 2.5 186 ± 5.8 133 ± 15.0 

10,000 84. 7.8 ,,. 2,7 106 ± 4.2 266 ± 11.3 101 ± 2.0 184 ± S.3 117±11.3 

Trial summary Negative Ncgalive Ncgltive Equivocal Negative Negalive Negative 

Positive control 1.302 ± 75.0 198 ± 16.3 600 ± 43.4 952 ± 93.1 761 ± 42.0 311 ± 19.4 J09 ± 9.9 

I SlUdy perfonned II Microbiological Associates, IDe. 1lIe dcuilcd prorocol is presented in Zeiger tt af. (1988). 

b Revertants aTt presented IS mean ± ~tandl.rd cnor from three plates. 

t No standard error calcuJaled due to loss of replicate cultures 

d The positive controls iIllhe absence of metabolic activation were sodium uide (fAIS3S and TAIOOl, 9-aminoacridine (fA97 and 

TA1537), and 4-nifro-o.-pbenylenediamine (fA98). The positive control for ~tabolk activation with all strains was 2-amiDoanlhracene. 

c S1igbt IOdcit)' 
Genetic Toxit::ology 223 

TABLE E2 

Induction or Sister Chromatid Excbanges in Chinese Hamster Ovary Cells 

by Scopoiamine Hydrobromide Tribydrate" 

No, of SCE>! Relative 

Compound 

Do>' 

(}.Ig/mL) 

Total 

C,lIs 

Chromosomu 

No, of 

SCE> 

Chromosome SCEst 

Cell 

H" 

in UrdU 

Change or SCEs! 

Chromosomtb 

(%) 

-So 

Sumnwy: NcBanve 

Oimtmylsulfo:dde SO 1,049 381 0.36 7.' 26.0 

Mitomycin-C 0.1)005 SO 1,049 '" 0." 10.2 26.0 34.12 

0.0050 10 210 287 1.36 28.7 26.0 276.28 

Scopolamine hydrobromide trihydralt 

SO 1.048 401 0.38 8.0 26.0 5.35 

SO " SO 1,048 420 0.40 8.4 26.0 10.34 

'60 SO 1.051 401 0.38 8.0 26.0 5.05 

"" " 1,050 40' 0.38 8.1 26.0 6.20 

P_0.234c 

+S' 

Trial 1 

Summary: Weakly positive 

OimtthylsulfoJ.ide SO 1,049 368 0.35 7.4 26.0 

Cyclophosphamide 

0.1 SO 1.049 466 0.44 '.3 26.0 26.63 

0.' 10 210 228 1.08 22.8 26.0 209.49 

Sc.opolamint hydrobromide lrihydnue 

160 " 1,050 394 0.37 7.' 26.0 .... "0 

1,600 " " 

1,049 

1,050 

407 

401 

0.38 

0.38 

8.1 

8.0 

26.0 

26.0 

10.60 

8.86 

',000 " 1,047 471 0.44 '.4 26.0 28.23· 

P<O.ool 

Trial Tl 

Sumnwy: Positive 

Dimethylsulfolide SO 1,047 410 0.39 8.2 26.0 

Cyclopbosphamide 

0.1 SO 1,049 '76 0.54 11.5 26.0 40.22 

0.' 10 210 247 1.17 24.7 26.0 200.36 

Scopolamint hydrobromide trihydrate 

2,000 " 1.048 '" 0.51 10.8 26.0 31.34· 

3,000 

4,000 " " 

1,050 

1,050 

'24 

'50 

0.49 

0.52 

10.5 

11.1 

26.0 

26.0 

27.«· 

35.22· 

',000 " 1,049 '82 0.5$ 11.6 26.0 41.68· 

P<OJXlI 
II 

2M	 Scopolamine Hydrobromide Trihydrate, NTP TR 44S 

TABLE £2 

Induction of Sister Chromatid Exchanges in Chinese Hamster Ovary Cells 

by Scopolamine Hydrobromlde Trihydrate (ronlinued) 

Compound ""'. V<,/mL) 

Total 

C.1Is 

No. of 

Chromo­

$omeJ 

No. of 

SCEo 

SCEsJ 

Chromosome SC&I 

C•• 

U" 

in BrdU 

Relative 

Change of seEs! 

Chromosome 

('1.) 

+59 (continued) 

Trial3c 

Sumtnuy: Nelltive 

Dinx:lhyl$Ulfo:dck ", 1,050 '10 0.39 8.2 26.0 

Cyclophosphamide 

0.1 ", 1,049 S30 0.'" 10.6 26.0 29.39 

0.• 10 211 237 1.12 23.7 26.0 181.66 

Scopolamine hydrobromide trihydralC 

2.000 

3.000 

", 

", 

1.048 

1,050 .,. 

'" 0.45 

0.43 

•••

'.1 

26.0 

26.0 

17.05 

10.73 

'.000 ", 1,049 '" 0.40 8.S 26.0 '.25 

S.OOO ", 1,051 m 0.<45 '.S 26.0 16.23 

P"'O.135 

•	 Positive resporue (:i!:205 increase over solvent control) 

•	 Srudy performed. It Environmental Health Research &: Testing, Inc. A detailed d~ription of the prolOCOl is pruerutd in Galloway 

n 01. (1987). SCE-siSler chromatid exchall&e; BrdU-bromodcoxyul1dine. 

SCEs/chromosome in treated cells venus SeE/chromosome in solvent control cells 

e Signi(1CUICe of SCEsichromo$OIDe tested by the linear regression lrend leA vs. 101 of !be dose 

d AIlered pH of lIledium .1 all doses in this trial 

e pH of lIledium adjuSied in this Ilia! with HEPES buffer 
Genetic ToxicolOJY 115 

TABLE E3 

Induction of Chromosomal Aberrations in Chinese Hamster Ovary Cells 

by Scopolamine Hydrobromide Trihydratea 

Do" 

lPg/mLl 

Total 

Cells 

-S9 

No. or 

AM 

Absl 

C•• 

Cells with 

Abs ('1» 

Do" 

lPg/mL) 

Total 

C.1Is 

+89 

No. or 

Ab, 

Absl 

C•• 

Cells with 

Abs ('1» 

Harvest time: 12.S bouTS 

Sununaly: Negative 

Trial 1 - Harvest time: I3.S bouTS 

Summary: Weak Positive 

Dimethylsulfoxide 

200 2 0.01 1.0 

Dimethylsulfoxide 

200 0.01 0.' 

Mitomycin.(: 

0.062S 

0.2SOO 

200 

50 

'"

17 

0.20 

0.34 

16.0 

30.0 

Cyclophosphamide 

2.' 

7.' 

200 

50 

"

23 

0.18 

0.46 

IS.O 

36.0 

Scopolamine hydrobromide trihydrate 

1.000 200 1 

1.600 200 2 

3.000 200 I 

0.01 

0.01 

0.01 

0.' 

1.0 

0.' 

Scopolamine bydrobromide trilIydrate 

1.600 200 3 

3.000 200 2 

'.000 200 12 

0.., 

0.01 

0.06 

U 

1.0 

6.0· 

P.0.6SSb P<O.OOI 

Trial ZC • HarveR time: 13.0 boul'S 

Summary: weak positive 

Dimethylsulfoxide 

200 0.01 0.' 

Cyclopbosph.lmide 

2.' 

7.' 

200 

50 

28 

17 

0.14 

0.34 

12.0 

34.0 

ScopolamillC hydrobromide trihydrate 

1.600 200 3 

3.000 200 2 

'.000 200 28 

0.., 

0.01 

0.14 

U 

1.0 

11.0 

P<O.OOI 

• Positive (P<0.05) 

a Study performed al Environmcnu.1 Health Research &. Testinl. tnc. The deu.iJed protocol is p~$(nted in Galloway tt al. (1987). 

Abs"'abcrntions. . 

b Signific2nce of perceru cells with aberrations !Wed by !be linear regression uend test vs. klg of the dose 

, pH of culture medium adju.sred with HEPES buffer 
226 Scopolamine Hydrobl'"omide Trihydrate, NTP TR 445 

TABlE E4 

Frequency or Micronuclei in Mouse Peripheral Blood Erytbrocytes Following Treatment 

wilh Scopolamine Hydrobromide Trihydrate by GaYage for 14 Weeks" 

Dos< l\{icronuckated Number 

(mglkg) NCEslI,OOO CtUsb or Mice 

....k 

0 

15 

" '" 

1,200 "'" 

Trend lest 

Female 

0 

IS 

13S 

"

400 

1.200 

Trend lest 

1.95 ± 0.14 

2.12 ± 0.06 

2.18 ± 0.20 

2.22 ± 0.12 

1.91 ± 0.21 

2.24 ± 0.21 

1.59 ± 0.09 

1.43 ± 0.11 

1.64 to.14 

1.43 ± 0.09 

1.40 ± 0.12 

1.38 ±O.12 

P-O.1219 

10 

JO 

10 

10 

10 

10 

10 

10 

10 

JO 

JO 

10 

• Slides scorut al United SlateS Ocpanmenl of AlricuID.lTt. The detailed prococol is presented by MacGregor n ai. (1990); 10,000 NeEs 

scored per animal. 

b Dau pruenlC:d &5 mean ± SlI.ndard elTOr of the mean. NeE _ nonnochronadc erythrocyte. 

e SignifICance of micronuclealC:d NeEs determined by • one-lailed Coeh~-Annita&c trend test. 
227 

APPENDIX F 

ORGAN WEIGHTS 

AND ORGAN-WEIGHT-TO-BODY-WEIGHT RATIOS 

TABLEF1 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats 

in the 16-Day Gavage Study of Scopolamine Hydrobromide Trihydrate . . . . . . . . . . 228 

TABLEF2 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats 

in the 14-Week Gavage Study of Scopolamine Hydrobromide Trihydrate . . . ... . .. 230 

TABLEF3 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats 

at the 15-Month Interim Evaluation in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate .. . . ... . . . . . . . . . . . . . . . . . . . . . . 232 

TABLE F4 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice 

in the 16-Day Gavage Study of Scopolamine Hydrobromide Trihydrate . . . . . . . . . . 233 

TABLEF5 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice 

in the 14-Week Gavage Study of Scopolamine Hydrobromide Trihydrate . . . . . . . . . 235 

TABLE F6 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice 

at the 15-Month Interim Evaluation in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydrate . . . . . .. . . . . . . . .. . . . .... . . . . . 237 
228 Scopolamine Hydrobromide Triiydrate, NTP TR 445 

TABLEF1 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 16-Day Gavage Study 

of Scopolamine Hydrobromide Trihydratea 

Vehicle 

Control 75 mgkg 150 mgkg 300 mgkg 600mgkg, 1,200 mgkg 

Male 

n 5 5 5 5 5 5 

Necropsy body wt 201 f 2 189 f 5 189 f 7 185 f 4 177 f 7** 179 f 5** 

Brain 

Absolute 1.804 f 0.015 1.760 f 0.016 1.748 f 0.053 1.738 f 0.021 1.722 f 0.036 1.708 f 0.022 

Relative 8.98 f 0.10 9.35 f 0.20 9.27 f 0.20 9.38 f 0.13 9.75 f 0.31 9.58 f 0.22 

Heart 

Absolute 0.724 f 0.027 0.648 f 0.029 0.664 f 0.028 0.640f 0.016 0.628 f 0.025 0.616 f 0.024* 

Relative 3.60 f 0.14 3.43 f 0.08 3.52 f 0.08 3.45 f 0.05 3.54 f 0.08 3.45 f 0.13 

R.Kidney 

Absolute 0.910 f 0.036 0.838 f 0.012 0.794 f 0.030* 0.848 f 0.014 0.816 f 0.031 0.820 f 0.038 

Relative 4.53 f 0.17 4.45 f 0.08 4.21 f 0.10 4.58 f 0.11 4.60 f 0.07 4.59 f 0.14 

Liver 

Absolute 9.976 f 0.354 9.252 f 0.296 10.128 f 0.565 8.774 f 0.231 9.510 f 0.763 8.912 f 0.382 

Relative 49.62 f 1.69 49.02 f 0.56 53.62 f 2.36 41.39 f 1.48 53.34 f 2.51 49.83 f 1.10 

Lung 

Absolute 0.996 f 0.039 1.024 f 0.122 1.122 f 0.198 0.922 f 0.027 0.932 f 0.061 0.862 f 0.051 

Relative 4.95 f 0.17 5.39 f 0.52 5.88 f 0.87 4.98 f 0.17 5.28 f 0.36 4.81 f 0.16 

R. Testis 

Absolute 1.112 f 0.038 1.063 f 0.011 4.058 f 0.035 1.065 f 0.066 1.054 f 0.046 1.05 1 f 0.052 

Relative 5.54 f 0.20 5.65 f 0.15 5.61 f 0.14 5.73 f 0.29 5.95 f 0.21 5.87 f 0.16 

Thymus 

Absolute 0.445 f 0.024 0.380 f 0.021 0.424 f 0.051 0.401 f 0,026 0.343 f 0.024* 0.330 f 0.013* 

Relative 2.21 f 0.12 2.01 f 0.07 2.22 f 0.20 2.16 f 0.11 1.93 f 0.07 1.85 f 0.09 
Organ Weight Analyses 229 

TABLEF1 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 16-Day Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle 

Control 75 mgkg 150 mg/kg 300 mgkg 600 mgkg 1,200 mgkg 

Female 

n 5 5 5 5 5 5 

Necropsy body wt 145 f 5 135 f 3 139 f 4 141 f 5 145 f 4 135 f 2 

Brain 

Absolute 1.712 f 0.010 1.662 f 0.026 1.632 f 0.029* 1.656 f 0.024 1.638 f 0.012 1.648 f 0.004 

Relative 11.84 f 0.33 12.28 f 0.12 11.75 f 0.13 11.82 f 0.38 11.35 f 0.32 12.24 f 0.17 

Heart 

Absolute 0.540 f 0,014 0.488 f 0.015 0.504 f 0.012 0.538 f 0.010 0.530 f 0.021 0.508 f 0.025 

Relative 3.73 f 0.08 3.60 f 0.08 3.64 f 0.13 3.84 f 0.12 3.66 f 0.05 3.77 f 0.19 

R. Kidney 

Absolute 0.662 f 0.016 0.606 f 0.023 0.632 f 0.018 0.652 f 0.021 0.688 f 0.024 0.650 f 0.013 

Relative 4.57 f 0.06 4.47 f 0.11 4.55 f 0.09 4.64 f 0.09 4.75 f 0.11 4.83 f 0.14 

Liver 

Absolute 6.270 f 0.224 5.790 f 0.199 6.168 f 0.209 6.334 f 0.298 6.840 f 0.444 6.194 f 0.265 

Relative 43.21 f 0.41 42.75 f 1.24 44.37 f 0.91 44.95 f 0.67 47.11 f 2.04 45.98 f 1.94 

Lung 

Absolute 0.776 f 0.037 0.688 f 0.024 0.846 f 0.050 0.788 f 0.056b 0.864 f 0.067 0.794 f 0.056 

Relative 5.34 f 0.15 5.08 f 0.12 6.11 f 0.43 5.61 f 0.21b 5.94 f 0.31 5.89 f 0.42

Thymus 

Absolute 0.369 f 0.018 0.336 f 0.017 0.362 f 0.021 0.345 f 0.021 0.346 f 0.014 0.314 f 0.014 

Relative 2.54 f 0.05 2.48 f 0.12 2.60 f 0.10 2.45 f 0.10 2.39 f 0.11 2.33 f 0.08 

* Significantly different (PSO.05) from the control group by Williams’ or Dunnett’s test 

** PSO.01 

a Organ weights and body weights are given in grams; organ-weight-to-body-weightratios are given as mg organ weightlg body weight 

(mean f standard error). 

b n-4 
230 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEF2 

of Scopolamine Hydrobromide Trihydratea 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 14-Week Gavage Study 

Control 

Vehicle 

15 mglkg 45 mgkg 135 mglkg 400 mgkg 1,200 mgkg 

Male 

n 10 9 10 8 4 lb 

Necropsybody wt 353 f 7 327 f 6** 314 f 4**309 f 6** 312 f 3** 32 1 

Brain 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Heart 

R.Kidney 

Liver 

Lung 

R. Testis 

Thymus 

0.89 f 0.05 

0.314 f 0.021 

4.04 f 0.08 

1.422 f 0.018 

4.28 f 0.15 

1.513 f 0.069 

40.90 f 0.82 

14.438 f 0.422 

3.86 f 0.05 

1.362 f 0.025 

2.96 f 0.03 

1.042 f 0.017 

5.68 f 0.12 

1.997 f 0.010 1.978 f 0.0151.922 f 0.013**1.900 f 0.020**1.883 f 0.024** 

6.05 f 0.09* ' 6.14 f 0.07**6.17 f 0.11** 6.03 f 0.10 

0.973 f 0.014**0.935 f 0.014** 0.904 f 0.016**0.898 f 0.005** 

2.98 f 0.06 2.98 f 0.022.93 f 0.03 2.87 f 0.03 

1.271 f 0.018** 1.211 f 0.026**1.170 f 0.015**1.280 f 0.038** 

3.89 f 0.08 3.86 f 0.05 3.80 f 0.064.10 f 0.13 

13.121 f 0.487* 12.188 f 0.192** 12.911 f 0.260* 13.420 f 0.347 

40.00 f 0.99 38.88 f 0.42 41.84 f 0.53 42.99 f 1.46 

1.451 f 0.0451.401 f 0.0511.381 f 0.0511.380 f 0.104 

4.45 f 0.184.48 f 0.19 4.48 f 0.15 4.42 f 0.36 

1.270 f 0.1451.441 f 0.0231.310 f 0.1071.425 f 0.043 

3.85 f 0.44 4.60 f 0.07 4.23 f 0.33 4.56 f 0.14 

0.247 f 0.011** 0.241 f 0.014** 0.216 f 0.012** 0.195 f 0.011** 

0.75 f 0.03* 0.77 f 0.04* 0.70 f 0.03** 0.62 f 0.03** 0.69 

0.221 

4.37 

1.405 

4.05 

1.300 

46.08 

14.800 

3.89 

1.250 

2.71 

0.870 

5.73 

1.840 

L 
Organ Weight Analyses 231 

TABLEF2 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 14-Week Gavage Study 

of Scopolamine Hydrobromide Trihydrate(continued) 

Vehicle 

Control 15 mgkg 45 mgkg 135 mgkg 400 mgkg 1,200 mgkg 

Female 

n 10 10 9 9 9 3 

Necropsy body wt 204 f 2 197 f 2* 198 f 1922* f 3** 192 f 2** 194 f 4* 

Brain 

Absolute 1.830 f 0.01 1 1.822 f 0.013 1.834 f 0.0161.782 f 0.011* 1.782 f 0.017* 1.723 f 0.027** 

Relative 8.98 f 0.07 9.26 f 0.09 9.26 f 0.10 9.30 f 0.10 9.28 f 0.09 8.87 f 0.06

Heart 

Absolute 0.676 f 0.012 0.685 f 0.009 0.693 f 0.022 0.679 f 0.023 0.630 f 0.012 0.653 f 0.013 

Relative 3.32 f 0.05 3.48 f 0.05 3.50 f 0.11 3.54 f 0.09 3.28 f 0.06 3.36 f 0.09 

R.Kidney 

Absolute 0.807 f 0.013 0.791 f 0.013 0.798 f 0.0120.788 f 0.023 0.757 f 0.022 0.833 f 0.026 

Relative 3.96 f 0.07 4.02 f 0.05 4.03 f 0.07 4.10 f 0.10 3.93 f 0.09 4.29 f 0.05

Liver 

Absolute 7.062 f 0.151 7.230 f 0.124 7.111 f 0.171 6.827 f 0.223 7.278 f 0.176 8.260 f 0.100** 

Relative 34.67 f 0.73 36.74 f 0.55 35.90 f 0.84 35.56 f 0.94 37.87 f 0.80** 42.52 f 0.44** 

Lung 

Absolute 1.115 f 0.038 1.026 rt 0.032 1.032 f 0.0311.011 f 0.038 1.014 f 0.024 1.010 f 0.040 

Relative 5.47 f 0.17 5.21 f 0.14 5.21 f 0.15 5.26 f 0.15 5.28 f 0.13 5.19 f 0.12

R. Ovary 

Absolute 0.057 f 0.007 0.058 f 0.002 0.053 f 0.0030.054 f 0.003 0.059 f 0.005 0.063 f 0.010 

Relative 0.28 f 0.03 0.29 f 0.01 0.27 f 0.01 0.28 f 0.02 0.31 f 0.02 0.32 f 0.05 

Thymus 

Absolute 0.266 f 0.011 0.229 f 0.005** 0.223 f 0.006** 0.213 f 0.009** 0.204 f 0.006** 0.171 f 0.003** 

Relative 1.31 f 0.05 1.17 f 0.02** 1.13 f 0.03** 1.11 f 0.04** 1.07 f 0.04** 0.88 f 0.01** 

Uterus 

Absolute 0.740 f 0.099 0.538 f 0.036 0.631 f 0.0760.743 f 0.086 0.687 f 0.084 0.453 f 0.020 

Relative 3.63 f 0.48 2.73 f 0.18 3.19 f 0.38 3.84 f 0.41 3.61 f 0.48 2.34 f 0.13 

* Significantly different (PS0.05) from the control group by Williams’ or Dunnett’s test 

** PSO.01 

a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios are given as mg organ weightlg body weight 

(mean f standard error). 

n= 1; no standard error calculated 
232 Scopolamine Hydrobromide "hydrate, NTP TR 445 

TABLEM 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats at the 15-Month Interim Evaluation 

in the 2-YearGavage Study of Scopolamine Hydrobromide Trihydratea 

Male 

n 10 10 10 10 

Necropsy body wt 493 f 9 492 f 11 481 f 10 448 f 9** 

R. Epididymis 

Absolute 0.423 f 0.022 0.464f 0.012 0.428f 0.014 0.441 f 0.017 

Relative 0.86f 0.04 0.94 f 0.02 0.89 f 0.03 0.99f 0.04* 

R. Kidney 

Absolute 1.614f 0.029 1.565 f 0.050 1.543 f 0.047 1.366 f 0.036** 

Relative 3.28 f 0.07 3.18f 0.06 3.20 f 0.06 3.06 f 0.08 

Liver 

Absolute 18.032f 0.477 17.953 f 0.579 17.382 f 0.580 15.319 f 0.481** 

Relative 36.65 f 0.83 36.46 f 0.71 36.11 f 0.81 34.21 f 0.84 

R. Testis 

Absolute 1.884f 0.229 1.680f 0.075 1.777 f 0.169 1.669 f 0.142 

Relative 3.83 f 0.47 3.43 f 0.18 3.67 f 0.30 3.76 f 0.37 

Female 

n 10 10 10 

Necropsy body wt 293 f 12 -b 285 f 5 274 f 7 

R. Kidney 

Absolute 0.938 f 0.017 - 0.869 f 0.014** 0.845 f 0.021** 

Relative 3.23 f 0.10 - 3.06 f 0.073.09 f 0.04 

Liver 

Absolute 9.198 f 0.322 - 8.869 f 0.147 8.392f 0.235* 

Relative 31.54f 0.92 - 31.19 f 0.43 30.70 f 0.59 

* Significantly f (PsO.05) from the control group by Williams' or Dunnett's test 

** PSO.01 

a Organ weights and body weights are given in grams; organ-weight-to-body-weightratios an given as mg organ weight/g body weight 

(mean f standard error). 

Female rats in the 1 mg1kg group wen not necropsied at the interim evaluation dueto high mortality. 
Organ Weight Analyses 233 

of Scopolamine Hydrobromide Trihydratea 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice in the 16-Day Gavage Study 

TABLEF4 

~ ~ ~ ~ ~~ ~ ~ ~ 

Control 

Vehicle 

150 mgkg 250 mgkg 450 mgkg 900 mgkg 1,800 mgkg 

Male 

n 5 5 5 5 5 4 

Necropsybody wt 21.5 f 0.7 22.1 f 0.5 21.1 f 0.6 21.6 f 1.1 22.0 f 0.4 22.0f 1.0 

Brain 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Relative 

Absolute 

Heart 

R. Kidney 

Liver 

Lung 

R. Testis 

Thymus 

20.92 f 0.68 

0.448 f 0.002 

5.14 f 0.27 

0.110f 0.004 

9.70 f 0.41 

0.208 f 0.007 

56.41 f 1.99 

1.212f 0.051 

4.62 f 0.22 

0.099f 0.004 

6.92f 0.36 

0.148 f 0.005 

2.81 f 0.17 

0.060f 0.003 

19.85f 0.39 

0.438 f 0.006 

5.35 f 0.14 

0.118 f 0.002 

9.59 f 0.25 

0.212f 0.007 

58.18 f 1.21 

1.286 f 0.041 

4.40 f 0.21 

0.097 f 0.004 

6.88 f 0.31 

0.152 f 0.007 

2.83 f 0.10 

0.063f 0.003 

21.24 f 0.64 

0.446 f 0.005 

5.37 f 0.52 

0.112 f 0.008 

10.05 f 0.97 

0.210f 0.015 

6.78 f OSb 

0.145 f O.OIOb 

62.30 f 3.61 

1.306f 0.053 

4.45 f 0.22 

0.093 f 0.004 

2.42 f 0.13 

0.051 f 0.003 

20.63 f 1.03 

0.442f 0.007 

9.36 f 0.45 

0.202 f 0.012 

5.23 f 0.34 

0.112 f 0.005 

60.26 f 1.98 

1.306 f 0.084 

6.72f 0.39 

0.144f 0.005 

2.65 f 0.13b 

0.059 f O.OOZb 

4.26 f 0.25 

0.091 f 0.002 

20.10f 0.65 

0.442 f 0.015 

8.84f 0.39 

0.194f 0.007 

4.64 f 0.46 

0.102 f 0.010 

62.20 f 1.27 

1.368 f 0.030 

6.73f 0.45 

0,148f 0.010 

4.38f 0.14 

0.096f 0.004 

2.89 f 0.11 

0.063 f 0.002 

19.55 f 1.01 

0.428f 0.014 

5.16f 0.50 

0.113 f 0.009 

9.38 f 0.63 

0.205 f 0.010 

6.97 f 0.76 

0.153f 0.015 

64.44 f 1.42* 

1.420f 0.095 

2.44 f 0.08 

0.054f 0.002 

4.37 f 0.30 

0.095 f 0.003 
234 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEF4 

Organ Weights and Organ-Weight-to-Body-Weight Ratiosfor Mice in the 16-Day Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle 

Control 150 mg/kg 250 mgkg 450 mgikg 900 mgikg 1,800 mglkg 

Female 

n 5 4 5 5 5 3 

Necropsy body wt 19.7 f 0.3 19.7 f 0.3 19.6 f 0.3 19.4 f 0.419.0 f 0.419.3 f 0.3 

Brain 

Absolute 0.456f 0.004 0.453 f 0.009 0.442 f 0.005 0,452f 0.004 0.432 f 0.005** 0.433 f 0.009* 

Relative 23.14 f 0.33 23.00f 0.43 22.53 f 0.52 23.36f 0.44 22.76 f 0.31 22.41 f 0.09 

Heart 

Absolute 0.104f 0.004 0.095 f 0.003 0.098 f 0.006 0.102 f 0.002 0.090 f 0.003* 0.090f O.OO0 

Relative 5.27 f 0.16 4.83 f 0.07 5.00 f 0.34 5.27 f 0.09 4.74 f 0.15 4.66 f 0.08 

R.Kidney 

Absolute 0.164 f 0.005 0.170 f 0.012 0.156 f 0.007 0.162 f 0.006 0.154f 0.007 0.153 f 0.003 

Relative 8.32 f 0.21 8.63 f 0.53 7.96 f 0.41 8.36 f 0.23- 8.09 f 0.21 7.93 f 0.04 

Liver 

Absolute 1.036f 0.026 1.160f 0.032 1.078 f 0.032 1.122 f 0.046 1.118f 0.039 1.157 f 0.041 

Relative 52.53 f 0.94 58.93 f 0.90* 54.86 f 1.16* 57.83 f 1.34** 58.81 f 1.23** 59.88f 2.60** 

Lung 

Absolute 0.142 f 0.006 0.130f 0.006 0.154f 0.015 0,146f 0.004 0.144 f 0.004 0.133 f 0.009 

Relative 7.20f 0.24 6.60 f 0.19 7.82 f 0.68 7.55 f 0.28 7.60f 0.28 6.89 f 0.35 

Thymus 

Absolute 0.075 f 0.003 0.071 f 0.004 0.079f 0.007 0.078 f 0.003 0.075 f 0.004 0.067 f. 0.006 

Relative 3.80 f 0.16 3.62f 0.22 4.05f 0.40 4.01 f 0.16 3.93 f 0.17 3.46 f 0.27 

* Significantly different(PgO.05)from the control group by Dunnett’s test 

** PSO.01 

a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios are given as mg organ weightlg body weight 

(mean f standard error). 

n=4 
Organ Weight Analyses 235 

TABLEF5 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice in the 14-Week Gavage Study 

of Scopolamine Hydrobromide Trihydratea 

Vehicle 

Control15 mgkg 45 mgkg 135 mgkg 400 mgkg mgkg 1,200

Male 

n 10 10 10 9 10 8

Necropsy body wt 33.8 f 1.2 30.8 f 0.4** 30.6 f 0.4** 30.1 f 0.6** 28.7 f 0.6** 29.2 f 0.7** 

Brain 

Absolute 0.458 f 0.004 0.459 f 0.003 0.449 f 0.013 0.452 f 0.005 0.459 f 0.005 0.450 f 0.003 

Relative 13.68 f 0.38 14.93 f 0.20* 14.70 f 0.46* 15.04 f 0.21** 16.05 f 0.39** 15.49 f 0.34** 

Heart 

Absolute 0.156 f 0.007 0.141 f 0.003*0.146 f 0.0030.136 f 0.003**0.134 f 0.003**0.134 f 0.006** 

Relative 4.63 f 0.15 4.58 f 0.08 4.77 f 0.07 4.51 f 0.14 4.68 f 0.14 4.58 f 0.17 

R.Kidney 

Absolute 0.314 f 0.007 0.292 f 0.006* 0.282 f 0.007** 0.263 f 0.007**0.262 f 0.007**0.258 f 0.005** 

Relative 9.35 f 0.22 9.49 f 0.139.21 f 0.198.76 f 0.29 9.14 f 0.238.85 f 0.18 

Liver 

Absolute 1.621 f 0.076 1.541 f 0,055 1.490 f 0.040 1.473 f 0.049 1.426 f 0.051 1.474 f 0.075 

Relative 47.94 f 1.12 50.01 f 1.41 48.68 f 1.03 48.87 f 1.18 49.56 f 1.05 50.43 f 1.86 

Lung 

Absolute 0.187 f 0.010 0.160 f 0.005 0.177 f 0.006 0.186 f 0.015 0.170 f 0.009 0.171 f 0.010 

Relative 5.56 f 0.25 5.20 f 0.16 5.78 f 0.17 6.14 f 0.45 5.95 f 0.36 5.86 f 0.29 

R. Testis 

Absolute 0.11 1 f 0.002 0.116 f 0.002 0.118 f 0.002 0.112 f 0.003 0.111 f 0.002 0.109 f 0.002 

Relative 3.31 f 0.10 3.77 f 0.05** 3.85 f 0.05** 3.71 f 0.10** 3.86 f 0.08** 3.75 f 0.10**

Thymus 

Absolute 0.038 f 0.002 0.036 f 0.002 0.035 f 0.002 0.035 f 0.001 0.036 f 0.002 0.036 f 0.002 

Relative 1.14 f 0.04 1.16 f 0.06 1.14 f 0.08 1.15 f 0.03 1.25 f 0.05 1.22 f 0.07 
236 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEFJ 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice in the 14-Week Gavage Study 

of Scopolamine Hydrobromide Trihydrate (continued) 

Vehicle 

Control IS mgkg 45 mgkg 135 mgkg 400 mgkg 1,200 mgkg 

Female 

n 10 10 10 10 10 9 

Necropsy body wt 29.2 f 0.5 27.6 f 0.4' 27.5 f 0.5* 26.6 f 0.5** 26.3 f OS** 25.5 f 0.3** 

Brain 

Absolute 0.471 f 0.003 0.475 f 0.005 0.470 f 0.003 0.473 f 0.006 0.462 f 0.006 0.452f 0.003** 

Relative 16.15 f 0.23 17.23 f 0.18** 17.13 f 0.34** 17.84f 0.32** 17.62f 0.29** 17.72 f 0.13** 

Heart 

Absolute 0.131 f 0.004 0.129f 0.005 0.127f 0.003 0.128f 0.004 0.125 f 0.002 0.114f 0.004** 

Relative 4.49f 0.15 4.67 f 0.15 4.62 f 0.10 4.82 f 0.14 4.78 f 0.12 4.49 f 0.15 

R. Kidney 

Absolute 0.216 f 0.003 0.211 f 0.004 0.200 f 0.010 0.216 f 0.005 0.200f 0.006 0.190f 0.004** 

Relative 7.41 f 0.14 7.65,f 0.13 7.26 f 0.33 8.14f 0.19 7.62 f 0.23 7.44 f 0.13 

Liver 

Absolute 1.473 f 0.042 1.374 f 0.028 1.402 f 0.058 1.389 f 0.051 1.383 f 0.050 1.393 f 0.042 

Relative 50.38 f 0.97 49.83f 0.96 50.78 f 1.28 52.44f 2.26 52.58 f 1.34 54.50f 1.20 

Lung , 

Absolute 0.173 f 0.007 0.168 f 0.007 0.175f 0.007 0.180f 0.006 0.172 f 0.005 0.170 f 0.005 

Relative 5.95 f 0.31 6.08 f 0.18 6.37f 0.25 6.81 f 0.30* 6.56 f 0.19* 6.66 f 0.19 

R. Ovary 

Absolute 0.012 f 0.001 0.013f 0.001 0.011 f 0.001 0,012f 0.001 0.012f 0.001 0.010f 0.001 

Relative 0.42f 0.02 0.46 f 0.02 0.41 f 0.05 0.45 f 0.02 0.44 f 0.03 0.38 f 0.03 

Thymus 

Absolute 0.053 f 0.003 0.052f 0.002 0.046 f 0.002 0.052 f 0.001 0.049 f 0.003 0.044 f 0.002** 

Relative 1.82 f 0.09 1.87 f 0.04 1.68f 0.06 1.97 f 0.05 1.86 f 0.09 1.73 f 0.08 

UteNS 

Absolute 0.167f 0.017 0.119 f 0.010 0.153 f 0.016 0.156 f 0.016 0.127 f 0.013 0.121 f 0.015 

Relative 5.72 f 0.56 4.32f 0.35 5.55 f 0.54 5.85 f 0.56 4.85 f 0.50 4.72 f 0.56 

* Significantly different (PSO.05)fromthecontrol group by Williams' or Dunnett'stest 

** PSO.01 

a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios are given as mg organ weight/g body weight 

(mean f standarderror). 
Organ Weight Analyses 237 

TABLEF6 

Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice at the 15-Month Interim Evaluation 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydratea 

Male 

n 9 10 10 10 

Necropsy body wt 50.2 f 0.6 51.9 f 1.1 49.6 f 1.8 39.0 f 0.6** 

R. Epididymis 

Absolute 0.062 f 0.003 0.062 f 0.003 0.059 f 0.002 0.057 f 0.003 

Relative 1.23 f 0.05 1.19 f 0.05 1.20 f 0.06 1.45 f 0.06* 

R.Kidney 

Absolute 0.376 f 0.015 0.383 f 0.009 0.360 f 0.012 0.325 f 0.006** 

Relative 7.47 f 0.22 7.38 f 0.10 7.31 f 0.22 8.32 f 0.09** 

Liver 

Absolute 2.336 f 0.122 2.627 f 0.196 2.374 f 0.258 2.152 f 0.353 

Relative 46.54 f 2.34 50.28 f 3.05 47.39 f 3.96 55.43 f 9.41 

R. Testis 

Absolute 0.119 f 0.003 0.123 f 0.004 0,125f 0.002 0.117 f 0.003 

Relative 2.37 f 0.05 2.38 f 0.05 2.55 f 0.08* 2.99 f 0.04** 

Female 

n 10 10 10 10 

Necropsy body wt 53.2 f 1.9 50.6 f 2.0 48.2 f 1.5 40.7f 1.7** 

R.Kidney 

Absolute 0.256 f 0.005 0.250 f 0.006 0.241 f 0.004 0.237 f 0.008 

Relative 4.83 f 0.10 4.99f 0.14 5.04 f 0.13 5.87 f 0.16** 

Liver 

Absolute 1.915 f 0.058 1.858 f 0.073 1.825 f 0.064 1.820 f 0.051 

Relative 36.09 f 0.58 36.80 f 0.56 37.90 f 0.62 45.15 f 1.50** 

* Significantly different (PSO.05)from the control group by Williams’ or Dunnett’s test 

** PSO.01 

a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios am given as mg organ weightlg body weight 

(mean f standard error). 
238 Scopolamine Hydrobromide Trihydrate, NTP TR 445 
239 

APPENDIX G 

HEMATOLOGY RESULTS 

TABLE G1 Hematology Data for Rats in the 14-Week Gavage Study 

of Scopolamine Hydrobromide Trihydrate .......................... 240 

TABLE 62 Hematology Data for Rats at the 15-Month Interim Evaluation 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........ 241 

TABLE63 Hematology Data for Mice in the 14-Week Gavage Study 

of Scopolamine Hydrobromide Trihydrate .......................... 242 

TABLE 64 Hematology Data for Mice at the 15-Month Interim Evaluation 

in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate ........ 243 
240 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEG1 

Hematology Data for Rats in the 14-WeekGavage Study of Scopolamine Hydrobromide Trihydratea 

Vehicle Control 15 mgkg 45 mgikg 135 mgkg 400 rngikg 1,200 mgkg 

Male 

n 10 9 10 8 4 lb 

Hematocrit (%) 46.8 f 0.5 46.2 f 0.4 48.0 f 0.3* 48.1 f 0.3* 49.0 f 0.4** 48.9 

Hemoglobin (gldL) 15.2 f 0.2 15.3 f 0.2 15.7 f 0.1 15.9 f 0.1* 16.2 f 0.1** 16.4 

Reticulocytes (106/pL) 

Erythroctes (106/pL) 

0.50 f 0.03 

9.39 f 0.10 

0.47 f 0.02 

9.38 f 0.10 

0.52 f 0.03 

9.63 f 0.08 

0.42 f 0.03 

9.61 f 0.05 

0.40 f 0.04 

9.72 f 0.11 

0.46 

9.58 

Nucleated erythrocytes 

(10’1~~) 0.03 f 0.01 0.04 f 0.01 0.01 f 0.01 0.02 f 0.01 0.02 f 0.02 0.08 

Mean cell volume (&) 49.9 f 0.4 49.1 f 0.1 50.0 f 0.2 50.0 f 0.3 50.5 f 0.3 51.O 

Mean cell hemoglobin 

(PPI

Mean cell hemoglobin 

16.2 f 0.3 16.3 f 0.1 16.3 f 0.1 16.6 f 0.1 16.7 f 0.1 17.1 

concentration (g/dL) 32.6 f 0.5 33.2 f 0.1 32.8 f 0.2 33.1 f 0.2 33.1 f 0.1 33.5 

Platelets (10~1~~) 633.4 f 20.8 638.8 f 10.6 655.8 f 28.2’ 620.6 f 20.6 621.5 f 21.3 601.o 

Leukocytes (103/pL) 7.35 f 0.44 6.96 f 0.31 8.01 f 0.36 7.59 f 0.56 6.73 f 0.57 8.40 

Segmented neutrophils 

(1 031p~) 1.14 f 0.13 1.89 f 0.28* 2.07 f 0.17** 1.82 f 0.20** 1.90 f 0.12* 2.69 

Lymphocytes (103/pL) 5.65 f 0.35 4.68 f 0.34 5.67 f 0.41 5.37 f 0.37 4.65 f 0.45 5.38 

Atypical lymphocytes 

(1o~/~L) 0.12 f 0.03 0.08 f 0.03 0.04 f 0.02 0.08 f 0.05 0.00 f 0.00 0.00 

Monocytes (lO’lpL) 0.35 f 0.09 0.22 f 0.06 0.17 f 0.05 0.20 f 0.04 0.07 f 0.04* 0.34 

Eosinophils (IdlpL) 0.08 f 0.03 0.09 f 0.03 0.06 f 0.01 0.11 f 0.04 0.08 f 0.02 0.00 

Female 

n 9 10 9 9 8 3 

Hematocrit (%) 46.9 f 0.4 47.4 f 0.6 48.1 f 0.4 47.7 f 0.4 49.0 f 0.4** 49.1 f 0.9* 

Hemoglobin (gldL) 15.6 f 0.1 15.5 f 0.2 15.9 f 0.1 15.7 f 0.2 16.2 f 0.1** 16.4 f 0.3* 

Erythrocytes (106/pL) 8.82 f 0.07 8.88 f 0.11 9.09 

*0.07“ 8.98 f 0.10 9.17 f 0.08* 9.26 f 0.17* 

Reticulocytes (106/pL) 0.41 f 0.02 0.38 f 0.02 0.42 f 0.03 0.43 f 0.03 0.43 f 0.03 0.42 f 0.05 

Nucleated erythrocytes 

(10~1~~) 0.02 f 0.01 0.05 f 0.01 0.03 f 0.02 0.05 f 0.02 0.05 f 0.02 0.07 f 0.07 

Mean cell volume (&) 53.1 f 0.3 53.2 f 0.2 52.7 f 0.2 53.1 f 0.2 53.5 f 0.3 53.0 f 0.0 

Mean cell hemoglobin 

(Pg) 17.7 f 0.1 17.4 f 0.1 17.5 f 0.1 17.5 f 0.1 17.7 f 0.1 17.7 f 0.1 

Mean cell hemoglobin 

concentration (gldL) 33.3 f 0.2 32.6 f 0.1* 33.1 f 0.1 33.0 f 0.2 33.1 f 0.1 33.5 f 0.2 

Platelets (10~1~~) 759.6 f 39.6 764.3 f 31.7 758.4 f 27.5 807.4 f 29.8 810.1 f 44.3 698.7 f 14.6 

Leukocytes (IO’lpL) 6.37 f 0.33 6.23 f 0.40 6.21 f 0.43 8.57 f 0.38* 7.39 f 0.59 5.97 f 0.91 

Segmented neutrophils 

(103/p~) 1.45 f 0.11 1.30 f 0.16 1.45 f 0.12 2.16 f 0.29 1.73 f 0.17 1.68 f 0.57 

Lymphocytes (103/pL) 4.63 f 0.36 4.68 f 0.25 4.48 f 0.42 6.15 f 0.25* 5.35 f 0.48 4.10 f 0.38 

Atypical lymphocytes 

0.09 f 0.04 0.03 f 0.03 0.02 f 0.01 0.04 f 0.04 0.04 f 0.03 0.04 f 0.02 

Monocytes (lO’lpL) 0.14 f 0.05 0.15 f 0.04 0.18 f 0.06 0.15 f 0.07 0.20 f 0.08 0.08 f 0.03 

Eosinophils (103/pL) 0.06 f 0.01 0.07 f 0.02 0.07 f 0.02 0.07 f 0.03 0.04 f 0.01 0.07 f 0.04 

* Significantly different (PSO.05) from the control group by Dunn’s or Shirley’s test 

** PSO.01 

a Mean f standarderror.Statisticaltests wereperformedonunroundeddata. 

n=l; no standard error calculated 

n=9 
Hematology 

TABLE62 

Hematology Data for Rats at the 15-Month Interim Evaluation in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydratea 

Male 

n 

Hematocrit (%) 

Hemoglobin (g/dL) 

Erythrocytes(106/pL) 

Reticulocytes (106/pL) 

Nucleated erythrocytes (103/pL) 

Mean cell volume (fL) 

Mean cell hemoglobin (pg) 

Mean cell hemoglobin 

concentration (g/dL) 

Platelets (10~1~~) 

Leukocytes (103/pL) 

Segmented neutrophils (103/pL) 

Lymphocytes (103/pL) 

Monocytes (IdlpL) 

Eosinophils (103/pL) 

Female 

n 

Hematocrit (%) 

Hemoglobin (g/dL) 

Erythrocytes(106/pL) 

Reticulocytes (106/pL) 

Nucleated erythrocytes (103/pL) 

Mean cell volume (fL) 

Mean cell hemoglobin (pg) 

Mean cell hemoglobin 

concentration (g/dL) 

Platelets (1031p~) 

Leukocytes (103/pL) 

Segmented neutrophils(103/pL) 

Lymphocytes (103/pL) 

Monocytes (103/pL) 

Eosinophils (103/pL) 

10 

45.5 f 0.5 

15.2 f 0.2 

9.61 f 0.12 

0.18 f O.Olb 

0.06 f 0.03 

47.5 f 0.4 

15.8 f 0.1 

33.6 f 0.2 

533.3 f 21.9 

4.75 f 0.18 

1.54 f 0.10 

3.09 f 0.14 

0.04 f 0.01 

0.08 f 0.02 

9 

44.2 f 0.7 

15.2 f 0.3 

8.67 f 0.13 

0.15 f 0.01 

0.03 f 0.01 

51.0 f 0.6 

17.6 f 0.1 

34.5 f 0.4 

499.9 f 30.4 

3.21 f 0.37 

1.07 f 0.24 

2.07 f 0.13 

0.01 f 0.01 

0.07 f 0.01 

10 

46.3 f 0.3 

15.3 f 0.0 

9.60 f 0.12 

0.16 f 0.01 

0.03 f 0.02 

48.2 f 0.6 

16.0 f 0.2 

33.1 f 0.2 

554.7 f 19.0 

5.35 f 0.30 

1.64 f 0.10 

3.65 f 0.25 

0.04 f 0.01 

0.03 f 0.01 

-C 

-

-

-

-

-

-

-

-

-

-

-

-

-

* Significantly different (P50.05) from the control group by Dunn’s or Shirley’s test 

** P50.01 

a Mean f standard error. Statisticaltestswereperformedonunroundeddata. 

n=9 

Female rats in the 1 mg/kg group were not necropsied at the interim evaluation dueto high 

10 

47.1 f 0.8 

15.4 f 0.2 

9.67 f 0.18 

0.18 f 0.01 

0.01 f 0.01 

48.6 f 0.8 

16.0 f 0.2 

32.8 f 0.3 

555.8 f 23.4 

5.13 f 0.29 

2.03 f 0.27 

2.94 f 0.12 

0.03 f 0.01 

0.12 f 0.03 

10 

44.7 f 0.4 

15.4 f 0.1 

8.84 f 0.05 

0.13 f 0.01 

0.04 f 0.02 

50.7 f 0.4 

17.5 f 0.1 

34.5 f 0.3 

527.9 f 23.4 

3.23 f 0.31 

1.11 f 0.14 

2.05 f 0.21 

0.01 f 0.01 

0.05 f 0.01 

mortality. 

10 

48.5 f 0.8+* 

15.8 f 0.2 

9.91 f 0.14 

0.19 f 0.01 

0.03 f 0.02 

48.9 f 0.5 

16.0 f 0.1 

32.7 f 0.2* 

537.5 f 43.0 

4.32 f 0.24 

1.18 f 0.12 

2.98 f 0.16 

0.03 f 0.01 

0.13 f 0.03 

10 

45.1 f 0.5 

15.6 f 0.2 

8.93 f 0.09 

0.11 f 0.01** 

0.02 f 0.01 

50.5 f 0.4 

17.4 f 0.1 

34.5 f 0.2 

583.2 f 20.5 

3.12 f 0.27 

1.01 f 0.11 

2.02 f 0.18 

0.02 f 0.01 

0.07 f 0.02 
242 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE63 

Hematology Data for Mice in the 14-Week Gavage Study of Scopolamine Hydrobromide Trihydratea 

~ ~ ~~~~~ 

Vehicle Control 15 mgkg 45 mgkg 135 mgkg 400 mgkg 1,200 mgkg 

Male 

n 10 9 10 9 10 8 

Hematocrit (%) 41.8 f 0.8 41.7 f 0.8 41.7 f 0.5 41.7 f 0.8 42.3 f 0.9 42.1 f 0.9 

Hemoglobin (gldL) 15.6 f 0.2 15.5 f 0.2 15.5 f 0.1 15.7 f 0.2 15.7 f 0.2 15.8 f 0.3 

Erythrocytes (106/pL) 10.25 f 0.17 10.05 f 0.13 10.19 f 0.10 10.21 f 0.14 10.06 f 0.16 9.95 f 0.21 

Mean cell hemoglobin 

Mean cell volume (a)

Reticulocytes (106/pL) 0.29 f 0.05 

40.8 f 0.4 

0.22 f 0.03 

41.4 f 0.4 

0.18 f 0.04 

40.9 f 0.3 40.8 f 0.4 

0.28 f 0.05 

42.2 f 0.3* 

0.23 f 0.06 

42.4 f 0.4* 

0.32 f 0.02 

Mean cell hemoglobin

(PP) 15.3 f 0.115.4 f 0.115.3 f 0.0 15.3 f 0.1 15.6 f 0.1* 15.9 f 0.2** 

concentration (gldL) 37.4 f 0.3 37.1 f 0.5 37.2 f 0.2 37.6 f 0.4 37.2 f 0.4 37.6 f 0.2 

Leukocytes (103/pL) 

Platelets (10~1~~) 1,105.1 f 28.3 

2.08 f 0.40 

1,141.6 f 55.2b 

2.46 f 0.33 

1,104.8 f 16.1 

3.10 f 0.38 3.30 f 0.66 

984.8 f 52.2 

3.61 f 0.73 

1,016.4 f 31.0 

2.94 f 0.50 

1,060.5 f 53.4 

Segmented neutrophils 

(10~1~~) 0.39 f 0.12 0.37 f 0.08 0.68 f 0.13* 0.78 f 0.12* 0.84 f 0.14* 0.77 f 0.12* 

Lymphocytes (103/pL) 1.67 f 0.29 2.05 f 0.27 2.33 f 0.25 2.45 f 0.54 2.61 f 0.55 2.11 f 0.39 

Atypical lymphocytes 

(10~1~~) 0.00 f 0.00 0.00 f 0.00 0.01 f 0.01 0.01 f 0.01 0.02 f 0.01 0.01 f 0.01 

Monocytes (1031pL) 0.01 f 0.01 0.01 f 0.01 0.03 f 0.03 0.02 f 0.02 0.08 f 0.04 0.04 f 0.02 

Eosinophils (103/pL) 0.01 f 0.01 0.02 f 0.01 0.04 f 0.01 0.03 f 0.02 0.07 f 0.02 0.01 f 0.01 

Female 

n 10 10 9 9 10 8 

Hematocrit (%) 41.3 f 0.8 41.2 f 0.7 42.1 f 0.4 42.2 f 0.7 42.2 f 0.7 40.8 f 0.5 

Hemoglobin (gldL) 15.3 f 0.2 15.4 f 0.2 15.5 f 0.1 15.8 f 0.2 15.7 f 0.2 15.5 f 0.1 

Erythrocytes (106/pL) 9.86 f 0.16 9.97 f 0.13 10.00 f 0.14 10.13 f 0.1010.20 f 0.15 9.80 f 0.09 

Reticulocytes (106/pL) 0.22 f 0.03 0.23 f 0.04 0.24 f 0.06 0.26 f 0.05 0.30 f O.OSb 0.20 f 0.06 

Mean cell volume (fL) 41.9 f 0.3 41.3 f 0.5 42.1 f 0.5 41.7 f 0.3 41.4 f 0.3 41.8 f 0.4 

Mean cell hemoglobin 

Mean cell hemoglobin

(Pg) 15.5 f 0.1 15.4 f 0.115.5 f 0.1 15.6 f 0.1 15.4 f 0.1 15.8 f 0.1 

concentration (gldL) 37.0 f 0.5 37.3 f 0.5 36.9 f 0.2 37.3 f 0.4 37.3 f 0.3 38.0 f 0.4 

Leukocytes (103/pL) 

Platelets (10~1~~) 

3.39 f 0.48 

955.8 f 45.8 979.1 f 37.8 

3.20 f 0.43 

983.8 f 32.5 

3.50 f 0.47 

1,080.0 f 35.3 

3.07 f 0.40 

975.0 f 39.4 

3.23 f 0.63 3.01 f 0.44 

977.3 f 80.5 

Segmented neutrophils 

(~o~/~L) 0.43 f 0.08 0.68 f 0.11 0.63 f 0.12 0.60 f 0.17 0.72 f 0.17 0.53 f 0.09 

Lymphocytes (lO’lpL) 2.89 f 0.41 2.41 f 0.30 2.78 f 0.38 2.39 f 0.32 2.42 f 0.48 2.41 f 0.35 

Atypical lymphocytes 

(10~1~~) 0.01 f 0.01 0.03 f 0.02 0.01 f 0.01 0.01 f 0.01 0.02 f 0.01 0.01 f 0.01 

Monocytes (103/pL) 0.03 f 0.01 0.02 f 0.01 0.03 f 0.02 0.06 f 0.03 0.05 f 0.03 0.02 f 0.01 

Eosinophils (103/pL) 0.03 f 0.01 0.06 f 0.02 0.03 f 0.01 0.01 f 0.01 0.02 f 0.01 0.04 f 0.01 

** P40.01 

* Significantly different (P40.05) from the control group by Dun” or Shirley’s test 

a Mean f standard error. Statisticaltestswereperformedonunroundeddata. 

n=9 
Hematology 243 

TABLE64 

Hematology Data for Mice at the 15-Month Interim Evaluation in the 2-Year Gavage Study 

of Scopolamine Hydrobromide Trihydratea 

Male 

n 

Hematocrit (%) 

Hemoglobin (g/dL) 

Erythrocytes (106/pL) 

Reticulocytes (106/pL) 

Nucleated erythrocytes (103/pL) 

Mean cell volume (a) 

Mean cell hemoglobin (pg) 

Mean cell hemoglobin 

concentration (gldL) 

Platelets (10~1~~) 

Leukocytes (103/pL) 

Segmented neutrophils (103/pL) 

Lymphocytes (103/pL) 

Monocytes (103/pL) 

Eosinophils (103/pL) 

Female 

n 

Hematocrit (%) 

Hemoglobin (gldL) 

Erythrocytes (106/pL) 

Reticulocytes (106/pL) 

Nucleated erythrocytes (103/pL) 

Mean cell volume (fL) 

Mean cell hemoglobin (pg) 

, Mean cell hemoglobin 

concentration (g/dL) 

Platelets (10~1~~) 

Leukocytes (103/pL) 

Segmented neutrophils (103/pL) 

Lymphocytes (103/pL) 

Monocytes (103/pL) 

Eosinophils (103/pL) 

10 

46.8 f 0.9 

14.7 f 0.2 

9.82 f 0.17 

0.15 f 0.01 

0.00 f 0.00 

47.6 f 0.5 

14.9 f 0.1 

31.4 f 0.3 

1,315 f 41 

7.69 f 0.70 

1.88 f0.22 

5.66 f0.48 

0.01 f0.01 

0.14 f0.04 

10 

48.4 f 0.4 

15.0 f 0.1 

9.96 f 0.08 

0.19 f 0.01 

0.00 f0.00 

48.6 f 0.3 

15.0 f 0.1 

31.0 f 0.2 

928 f 20 

4.84 f 0.50 

1.20 f 0.17 

3.57 f 0.37 

0.00 f 0.00 

0.07 f 0.03 

10 

47.7 f 1.0 

15.1 f 0.2 

10.23 f 0.24 

0.18 f 0.01 

0.01 f 0.01 

46.6 f 0.8 

14.8 f 0.2 

31.7 f 0.3 

1,216 f 59 

7.61 f0.77 

1.49 f 0.23 

5.98 f 0.53 

0.02 f0.02 

0.11 f 0.04 

10 

48.2 f 0.5 

15.0 f 0.2 

9.83 f 0.12 

0.18 f 0.01 

0.00 f 0.00 

49.0 f 0.3 

15.2 f 0.1 

31.0 f 0.2 

971 f 22b 

4.35 f 0.43 

1.01 f 0.08 

3.25 f 0.35 

0.00 f 0.00 

0.09 f 0.03 

10 

47.0 f 0.9 

14.8 f 0.2 

9.86 f 0.24 

0.16 f 0.01 

0.00 f 0.00 

47.8 f 0.5 

15.0 f 0.2 

31.4 f 0.3 

1,272 f 26 

7.14 f 0.47 

1.43 f 0.21 

5.45 f 0.41 

0.00 f 0.00 

0.26 f 0.06 

10 

48.4 f 0.3 

14.9 f 0.1 

9.88 f 0.10 

0.19 f 0.01 

0.00 f 0.00 

49.0 f 0.5 

15.1 f 0.1 

30.8 f 0.2 

1,005 f 57 

4.46 f 0.44 

1.09 f 0.13 

3.30 f 0.36 

0.00 f 0.00 

0.07 f 0.02 

10 

45.7 f 0.8 

14.4 f 0.2 

9.66 f 0.16 

0.18 f 0.03 

0.00 f 0.00 

47.3 f 0.6 

14.9 f 0.2 

31.5 f 0.3 

1,357 f 61 

5.88 f 0.52 

1.63 f 0.22 

4.15 f 0.33 

0.01 f 0.01 

0.09 f 0.02 

10 

46.5 f 0.4* 

14.5 f 0.1** 

9.57 f 0.07* 

0.17 f 0.01 

0.00 f 0.00 

48.5 f 0.4 

15.2 f 0.1 

31.2 f 0.2 

1,019 f 29 

5.10 f 0.46 

1.51 f 0.20 

3.48 f 0.34 

0.00 f 0.00 

0.11 f 0.01 

* Significantly different (P10.05) from the control group by DUM’S or Shirley’s test 

** PSO.01 

a Mean f standard error. Statisticaltestswereperformed on unroundeddata. 

n=9 
244 Scopolamine Hydrobromide Trihydrate,NTP TR 445 
AND ESTROUS CYCLE CHARACTERIZATION 

REPRODUCTIVE TISSUE EVALUATIONS 

APPENDIX H 

TABLEH2 

TABLEH1 

Summary of ReproductiveTissueEvaluationsandEstrousCycleCharacterization 

Summary of ReproductiveTissueEvaluationsandEstrousCycleCharacterization 

€orRats in the 14-WeekGavageStudy of Scopolamine Hydrobromide Trihydrate . . . 

for Mice in the 14-WeekGavageStudy of Scopolamine Hydrobromide Trihydrate ... 

246 

247 
246 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEH1 

Summary of Reproductive Tissue Evaluations and Estrous Cycle Characterization for Rats 

in the 14-Week Gavage Study of Scopolamine Hydrobromide Trihydratea 

Vehicle Control 45 mgkg . 135 mgkg 400 mgkg 

Male 

n 10 10 8 4 

Weights (g) 

Necropsy body wt 341 f 6 302 f 4** 300 f 7**b 304 f 3** 

R. cauda 0.196 f 0.004 0.196 f 0.007 .190 f 0.013 0.202 f 0.005

R. epididymis 0.417 f 0.004 0.427 f 0.005 0.394 f 0.026 0.424 f 0.006 

R. testis 1.422 f 0.018 1.441 f 0.023 1.310 f 0.107 1.425 f 0.043

Epididymal spermatozoal parameters 

Concentration 

(1OYg cauda epididymal tissue) 428.7 f 36.4 458.1 f 26.6 395.0 f 57.7 403.2 f 33.0

Motility (X) 77.94 f 1.10 76.10 f 1.65 67.09 f 9.85 76.93 f 0.99 

Abnormal sperm (%) 1.200 f 0.231 1.040 f 0.206 0.714 f 0.194' 1.100 f 0.265

Female 

n 10 9 9 9 

Necropsy body wt (g) 203 f 2 196 f 2* 190 f 2** 189 f 2** 

Estrous cycle length (days) 4.67 f 0.17d 5.11 f 0.26 4.43 f 0.20e 4.56 f 0.24 

Estrous stage (% of cycle) 

Diestrus 24.3 30.2 14.3 25.4 

Proestrus 20.0 15.9 19.0 15.9 

Estrus 34.3 34.9 42.9 34.9 

Metestrus 21.4 19.0 22.2 23.8 

Uncertain diagnoses 0.0 0.0 1.6 0.0 

* Significantly different (P10.05)from the control group by Williams' test ** P10.01 a Necropsy weights, organ weights, epididymal spermatozoal parameters, and estrous cycle length are presented as mean f standard 

error. Differences from the control group for organ weights, epididymal spermatozoal parameters, and estrous cycle stages are not 

significant by Dunn's test. By multivariate analysis of variance, exposed frmales did not differ significantly from the control females in 

relative length of time spent in the estrous stages. 

n=9

' n=7 

Estrous cycle longer t!an 12 days or unclear in 1 of 10 animals 

e Estrous cycle longer than 12 days or unclear in 2 of 9 animals 
Sperm Morphology and Vaginal Cytology Evaluations 247 

TABLEH2 

Summary of Reproductive Tissue Evaluations and Estrous Cycle Characterization for Mice 

in the 14-Week Gavage Study of Scopolamine Hydrobromide Trihydratea 

Male 

n 

Weights (9) 

Necropsy body wt 

R. cauda 

R. epididymis 

R. testis 

Epididymal spermatozoal parameters 

Concentration 

(106/g cauda epididymal tissue) 

Motility (%) 

Abnormal sperm (%) 

Female 

n 

Necropsy body wt (9) 

Estrous cycle length (days) 

Estrous stage (% of cycle)e 

Diestrus 

Proestrus 

Estrus 

Metestrus 

* 

Vehicle Control 

9 

32.7 f l.Ob 

0.017 f 0.001 

0.040 f 0.002 

0.111 f 0.002b 

10 

28.0 f 0.5 

4.00 f 0.00 

28.6 

20.0 

22.9 

28.6 

135 mgkg 400 mgkg 

9 10 

29.3 f 0.6** 28.7 + o.8** 

0.016 f O.OO0 0.015 f O.OO0* 

0.038 f 0.001 0.037 f 0.001 

0.112 f 0.003 0.111 f 0.002 

1,008.7 f 74.4 1,077.2 f 54.6 

74.31 f 0.82 75.30 f 1.14 

1.31 f 0.14 1.64 f 0.12 

10 10 

26.1 f 0.4** 26.2 f 0.5** 

4.20 f 0.13 4.00 f 0.00c 4.63 

22.9 20.0 

18.6 11.4 

34.3 45.7 

24.3 22.9 

Significantly different (PSO.05) from the control group by Williams’ test ** PSO.01 

1,200 mgkg 

28.9 f 0.6** 

0.015 f O.OOO** 

0.036 f 0.001 

0.109 f 0.002 

1,131.6 f 91.7 

76.66 f 1.14 

1.53 f 0.17 

9 

25.5 0.4** 

f o.18*d 

19.0 

15.9 

44.4 

20.6 

a Necropsy weights, organ weights, epididymal spermatozoal parameters, and estrous cycle length are presented as 

mean f standard error. Differences from the control group for epididymal spermatozoal parameters are not significant by DUM’Stest. 

n=10 

Estrous cycle longer than 12 days or unclear in 1 of 10 animals 

Estrous cycle longer than 12 days or unclear in 1 of 9 animals 

e Evidence shows that female mice exposed to 135 mglkg or 1,200 mglkg differed significantly (Wilk’s Crition, PSO.05) from the 

control females in the relative length of time spent in the estrous stages. 
248 Scopolamine Hydrobromide Trihydrate,NTP TR 445 
249 

APPENDIX I 

NEUROBEHAVIORAL STUDIES 

~~ODS........................................................... 250 

RESULTS ........................................................... 251 

TABLE11 NeurobehavioralDatafor Rats inthe2-YearGavageStudy 

ofScopolamineHydrobromideTrihydrate .... . . . . . .. . .. . . . . . . .. . . . . . .. 252 
250 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

NEUROBEHAVIORAL STUDIES 

METHODS 

Motor Activity: Motor activity was measured using a Figure 8 Photobeam Activity System (San Diego 

Instruments; San Diego, CA) with rearing detection attached. Measurements were made under relatively 

stress free conditions. Speakers placed next to the measurement chambers delivered white noise to mask 

noise which could have potentially affected behavior. Movement was measured as interruption of 

photobeams. Motor activity was determined over five 3-minute periods. Measurements of total activity 

were obtained and compensations were made for normal decreases in activity over the course of the 

session. 

Grip Strength: Forelimb and hindlimb grip strength were measured using a device similar to that 

described by Meyer et al. (1979), and data were entered directly into a Xybion@ electronic data collection 

system. Rats were allowed to grip a triangular ring with their forepaws and were pulled back along a 

platform until the grip was broken. As the backward motion continued, the rat's hindpaws reached a 

t-shaped hindlimb grip bar, which it was allowed to grasp and then forced to release by continued pulling. 

Chatillon push-pull strain gauges (Kew Gardens, NY) were used to record the maximum strain required to 

break the forelimb and hindlimb grip. Five trials were conducted on each rat with less than 1 minute 

between trials so that measure of degree of habituation or fatigue could be observed. The body weight of 

each rat was recorded at the end of the grip strength evaluation. 

Thermal Sensitivity: A model 550 Analgesia Meter (Omnitech Electronics, Inc.; Columbus, OH) was 

used for the test and data were entered directly into a Xybion@ system. The device consisted of a square 

acrylic arena with a clear acrylic cover mounted on a heat source. The rat was placed on the heat source 

and the arena was covered. The independent variable was rodent reaction time to the heat stimulus 

(55" C). The response most often observed was a vigorous licking of the hindpaws. Latency was 

measured manually by a built-in timer. Measurement began when the rat touched the plate and ended at 

the onset of the pain-sensing response. Rats failing to respond in 30 seconds were removed and assigned 

a maximum score of 30 seconds. 

Startle Responsiveness: Startle responsiveness was measured using an SR-LAB Startle Response System 

(San Diego Instruments; San Diego, CA), composed of four isolation chambers, a computer control unit 

and connection box, four startle chambers, and four test station control boxes. The computer controlled 

the presentation of all stimuli for four chambers simultaneously. The startle enclosure in which the test 

animal was placed was constructed of transparent acrylic and permitted testing of the animal with minimal 

restraint. The sensitivity of each enclosure could be individually and reproducibly adjusted. Response 

measurement took place within a sound-isolated cubicle equipped with a light, a ventilation fan, a small 

viewing port, and a small mini-max thermometer. 

Startle reflex was determined over 80 repeated trials. An acclimation period of 3 minutes was followed by 

20 startle trials with a tactile air puff (15 to 20 psi, 20 msec duration per trial). This was followed by 

40 prepulse trials in which an 80 to 90 dB(A) white noise prepulse preceded the tactile stimulus by 

100 msec. The final 20 trials were identical to the first 20 trials. Startle response for each rat for each 

trial was recorded following termination of the initial startle stimulus in order to avoid stimulus 

interference. With this method, the 20 msec wait time was added to all latencies. All trials were 

separated by an 8 second intertrial interval. A background noise level of 70 dB(A)was used. Each 

session took approximately 15 minutes to complete. The average response amplitude of each trial 

presentation was selected as the dependant variable for this study. 
Neurobehavioral Studies 251 

Passive Avoidance: Rats were tested using a passive avoidance system (San Diego Instruments; San 

Diego, CA) consisting of a computer control unit, control cabinets, and test enclosures. A computer unit 

controlled the presentation of stimuli for four chambers simultaneously and data were directly written to 

disk. The test enclosure was a two-compartment chamber, one brightly lit and the other dark. The 

lighted chamber was thesmallerofthetwo (5 in.wide X 7.75 in. high X 8 in. long). The darkened or 

shock chamber was 8 in. X 7.75 in. X 10 in. A guillotine-type door (Lafayette Instrument Co., 

Lafayette, IN) blocked a 3.5-inch square opening between the two chambers. A photocell was placed 

5 inches into the darkened area. The floor of the unit consisted of a series of 1/8-inch rods. The floor of 

the shock area consisted of 21 of these rods spaced 1/2 inch apart. Shock was delivered to the rat's feet 

through the rods by a solid state shockeddistributer (Coulbourn Instruments). 

To begin the training sessions, rats were placed in the smaller of the two enclosures, with the gate closed 

and the cue light off. A start/stop switch was pressed which began the timing of a 10-second adaptation 

period. At the end of the adaptation period, the cue light was illuminated, the gate was automatically 

opened and the timing of the session began. Rats were started on a staggered basis. When the subjects 

interrupted the photobeam in the shock compartment, the autogate closed and the shock (1 milliamp) was 

turned on for 3 seconds. The breaking of the photobeam indicated the end of the session. The maximum 

trial duration for the training sessions was 300 seconds. After shock had been administered, rats were 

immediately removed from the apparatus and returned to their respective cages. Retention test sessions 

were similar to the training sessions except that no shock was delivered when an animal broke the 

photobeam in the shock compartment. Maximum trial duration was 600 seconds, and an animal that 

stayed in the lighted compartment for the entire time received a maximum score of 600 seconds. 

' Passive avoidance trials were begun on day 90, followed 24 hours later by a retention testing session. 

Subsequent passive avoidance sessions (days 180, 270, 360, and 720) were limited to retention sessions. 

Due to the schedule of testing, animals administered scopolamine hydrobromide trihydrate prior to passive 

avoidance testing as well as prior to retention testing. This was different from protocols in which 

treatment was given immediately following training so that the animal was not performing the task for the 

first time while under the influence of an acute drug injection. A modification of the dosing and testing 

schedule was required to accommodate the gavage administration and retention test session during day 91 

for each group of animals since the retention test session (day 91) for a certain set of animals fell on the 

same day as the initial session (day 90) for another set of animals. This modification was needed because 

dosing was staggered from the beginning of the study. Animals tested on day 91 were gavaged at 

approximately the same time as they were on day 90. In addition, attempts were made to keep the time 

that the animals were placed into the apparatus on day 91 similar to the time that it occurred on the 

previous day. 

RESULTS 

Horizontal activity in males was not significantly altered by scopolamine administration; however, 

horizontal activity of 25 mg/kg females was significantly greater than that of the control group on days 90, 

180, and 360. Forelimb grip strength of 1 mg/kg males was significantly greater than that of the controls 

on day 360. In the 1 mg/kg female group, forelimb grip strength was significantly lower than that of the 

control group on day 90. Hindlimb grip strength of males and females was not significantly affected by 

scopolamine hydrobromide trihydrate administration. Pawlick latencies and startle responses of dosed 

males were not significantly different from those of the control group. In the 25 mg/kg female group at 

day 90, both pawlick latency and startle response were significantly lower than those of the control group. 

The startle response of 5 mg/kg females was also significantly lower than that of the control group. 

Passive avoidance in 25 mg/kg males was significantly lower than that of the control group at day 180; 

passive avoidance in females was not significantly affected by scopolamine hydrobromide trihydrate 

administration. 
252 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEI1 

Neurobehavioral Data €or Rats in the 2-YearGavage Study of Scopolamine Hydrobromide Trihydratea 

10 

114 f 4 

122 f 4 

350 f 8 

405 f 7 

452 f 7 

479 f 9 

575d 

151.2 f 6.2b 

126.5 f 7.9 

144.7 f 15.7 

50.0 f 11.3 

61.6 f 11.9 

45.4 f 9.1 

1.od 

31.2 f 4.7b 

30.4 f 4.3 

30.6 f 4.6 

11.5 f 4.0 

11.3 f 3.3 

5.0 f 1.3 

0.od 

0.433 f 0.011 

0.436 f 0.020 

1.440 f 0.015 

1.152 f 0.060 

1.310 f 0.066 

1.165 f 0.052* 

0.350d 

0.211 f 0.008 

0.205 f 0.007 

0.818 f 0.017 

0.745 f 0.030 

0.892 f 0.029 

0.779 f 0.029 

0.4SOd 

10 

113 f 4 

121 f 5 

326 f 7 

379 f 9 

429 f 14 

448 f 12 

443 f 24e 

131.3 f 11.4b 

107.2 f 10.7 

144.3 f 12.0 

77.9 f 13.1 

59.6 f 6.7 

58.8 f 10.2 

118.0 f ll.Oe 

23.8 f 4Sb 

15.7 f 2.7 

26.1 f 3.5 

10.6 f 4.0 

5.7 f 1.8 

4.5 f 1.2 

2.2 f 0.9e 

0.430 f 0.013 

0.410 f 0.019 

1.376 f 0.055 

1.049 f 0.050 

1.253 f 0.073 

0.966 f 0.037 

0.688 f 0.073e 

0.198 f 0.012 

0.201 f 0.012 

0.769 f 0.024 

0.724 f 0.031 

0.817 f 0.026 

0.742 f 0.026 

0.572 f 0.0251~ 

25 mgkg 

10 

117 f 4 

125 f 5 

318 f 6 

366 f 6 

403 f 8 

421 f lob 

420 f 15e 

127.0 f 9.8b 

131.1 f 14.0 

170.9 f 11.6 

98.8 f 6.8 

80.3 f 10.1 

91.8 f 18.3 

134.5 f 29.3e 

29.2 f 6.7b 

19.1 f 3.9 

29.5 f 4.7 

17.3 f 4.3 

5.8 f 2.5 

6.8 f 2,0b 

2.7 f l.le 

0.437 f 0.011 

0.403 f 0.015 

1.397 f 0.022 

1.162 f 0.044 

1.364 f 0.082 

0.998 f 0.040b 

0.625 f 0.064e 

0.209 f 0.012 

0.198 f 0.009 

0.784 f 0.023 

0.701 f 0.017 

03343 f 0.032 

0.759 f 0.044b 

0.637 f 0.037e 

Male 

n 

Body weight (g) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Vehicle Control 

10 

114 f 4 

122 f 4 

338 f 7 

385 f 9 

430 f 10 

455 f 10 

448 f 8' 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Total vertical activity count (15 minutes) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Forelimb grip strength (kg) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day.270 

Day 360 

Day 720 

Hindlimb grip strength (kg) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Total horizontal activity count (15 minutes) 

146.0 f 7.8b 

128.6 f 12.5 

171.0 f 13.3 

77.8 f 13.9 

71.5 f 7.1 

60.7 f 8.8 

92.6 f 19.4' 

27.7 f 5.3b 

26.3 f 4.8 

25.8 f 5.8 

13.9 f 3.2 

6.8 f 1.4 

5.3 f 1.8 

3.8 f 1.7' 

0.460 f 0.023 

0.448 f 0.017 

1.451 f 0.033 

1.124 f 0.044 

1.297 f 0.073 

0.995 f 0.059 

0.628 f 0.022' 

0.214 f 0.009 

0.200 f 0.008 

0.812 f 0.026 

0.775 f 0.029 

0.830 f 0.046 

0.771 f 0.030 

0.632 f 0.067' 
Neurobehavioral Studies 253 

TABLEI1 

Neurobehavioral Data for Rats in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate 

(continued) 

Male (continued) 

n 

Pawlick latency (seconds) 

Prestudy 

Day 1 

Day90 

Day180 

Day 270 

Day 360 

Day 720 

Startle response (milliseconds) 

Prestudy 

Day 1 

Day 90 

Day180 

Day 270 

Day360 

Day 720 

Passive avoidance (seconds) 

Day 90 

Day 91 

Day180 

Day 270 

Day360 

Day720 

Vehicle Control 

10 

5.48 f 0.24 

4.60 f 0.19 

6.07 f 0.45 

5.16 f 0.28 

3.86 f 0.34 

4.23 0.15 

4.26 f 0.27' 

43.80 f 5.66 

37.17 f 6.38 

33.97 f 5.55 

20.19 f 3.57 

17.99 f 1.90 

27.78 f 1.99 

28.75 f 4.32' 

41.91 f 29.13 

529.8 f 57.1 

557.7 f 52.7 

460.3 f 75.4 

485.4 f 80.6 

507.9 f 133.2' 

10 

6.20 f 0.67 

5.19 f 0.26 

6.98 f 0.67 

5.83 f 0.39 

4.10 f 0.20 

4.51 f 0.29 

5.90d 

35.73 f 5.28 

30.01 f 2.81 

32.76 f 3.45 

20.02 f 2.16 

13.99 f 1.24 

27.78 f 3.39 

22.04d 

21.91 f 5.41 

535.3 f 61.3 

558.6 f 49.8 

583.5 f 62.0 

563.9 f 60.1 

260.2d 

10 

5.83 f 0.29 

5.92 f 0.70 

5.71 f 0.36 

5.37 f 0.26 

3.96 f 0.20 

4.38 f 0.18 

5.05 f 0.46e 

44.03 f 8.66 

35.03 f 4.37 

31.29 f 3.86 

19.24 f 1.89 

14.28 f 1.72 

27.85 f 3.38 

35.74 f 11.93e 

59.36 f 33.54 

481.4 f 69.8 

564.8 f 30.7 

460.4 f 88.2 

561.7 f 50.6 

178.2 f 135.ge 

25 mgkg 

10 

5.27 f 0.25 

5.10 f 0.32 

6.47 f 0.60 

5.40 f 0.35 

4.09 f 0.29 

4.79 f 0.20b 

5.15 f 0.4ge 

46.97 f 5.80 

36.79 f 5.40 

32.18 f 3.08 

22.22 f 2.33 

17.81 f 1.66 

30.20 f 4.17b 

31.79 f 3.3ge 

23.12 f 6.17 

472.7 f 78.7 

355.6 f 87.0* 

488.9 f 77.3 

391.5 f llO.Ob 

373.1 f 151.6e 
254 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEI1 

Neurobehavioral Data for Rats in the 2-YearGavage Study of Scopolamine Hydrobromide Trihydrate 

(continued) 

Female 

n 

Body weights (g) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Vehicle Control 

10 

97 f 3 

105 f 3 

190 f 5 

207 f 3 

229 f 5 

262 f 6 

313 f log 

1 mgh 

10 

97 f 3 

105 f 3 

194 f 4 

211 f 4 

234 f 5 

261 f gb 

311 f 25g 

136.7f 16.1 

116.3 f 16.6 

155.8 f 10.6 

122.2f 17.9 

85.5 f 10.1 

101.2 f 14Ab 

72.3 f 15.9g 

23.3 f 4.3 

20.3 f 4.6 

31.9 f 4.9 

18.3 f 4.2 

14.7 f 3.8 

9.2f 1.4b 

3.5 f 2.1g 

0.456 f 0.013 

0.479f 0.017 

1.213 f 0.026* 

1.008 f 0.023 

0.993 f 0.054 

0.869 f 0.059b 

0.640 f 0.069g 

0.191 f 0.009 

0.193f 0.009 

0.690f 0.013 

0.635 f 0.022 

0.626f 0.019 

0.599 f 0.018b 

0.616 f 0.046g 

5 mgkg 

10 

97 f 3 

105 f 3 

188 f 3 

209 f 4 

227 f 7b 

255 f llb 

320 f 14g 

136.3 f 14.9 

117.0f 14.6 

148.5 f 10.3 

139.9 f 18.7 

103.0 f 16.6b 

89.2f 13.7b 

65.0f 10.2g 

21.1 f 4.8 

23.6f 3.5 

38.2f 8.3 

30.6 f 8.3 

22.7 f 6.6b 

12.4 f 3Sb 

1.5 f 1.2g 

0.404f 0.020 

0.453f 0.009 

1.183 f 0.020 

0.966f 0.034 

0.949f 0.028b 

0.872f 0.056b 

0.554f 0.046g 

0.172f 0.010 

0.187 f 0.012 

0.649 f 0.021 

0.651 f 0.030 

0.603 f0.018b 

0.612.f 0.021b 

0.563f 0.033g 

25 mgkg 

10 

98 f 3 

107 f 4 

188 f 2 

204 f 2b 

223 f 3b 

238 f gf 

295 f 19' 

134.9f 19.0 

151.7f 10.0 

204.7 f 14.8* 

178.0f 13.4b* 

117.9 f 14Sb 

134.6 f 14.3f* 

92.2 f 7.1' 

19.4f 3.9 

25.3 f 8.4 

27.1 f 5.2 

25.6 f 5.4b 

11.3 f 3.0b 

16.8 f 4.9f 

4.6 f 2.3' 

0.432 f 0.015 

0.430 f 0.023 

1.162f 0.021 

0.972 f. 0.032b 

0.922 f 0.049b 

0.878 f 0.041f 

0.625 f 0.084' 

0.183 f 0.007 

0.191 f 0.014 

0.647 f 0.020 

0.604 f 0.015b 

0.576 f 0.009b 

0.611 f 0.021f 

0.523 f 0.030' 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Total vertical activity count (15minutes) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Forelimb grip strength (kg) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Hindlimb grip strength (kg) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Total horizontal activity count (15 minutes) 

149.6f 14.8 

128.9f 15.2 

158.1 f 13.1 

115.2 f 17.9 

91.1 f 9.1 

80.5 f 8.1 

61.8 f 6.@ 

23.7 f 3.5 

21.2 f 3.7 

25.3 f 4.5 

16.0 f 4.1 

13.4 f 3.0 

1.7 f 1.8 

2.8 f l.lg 

0.455f 0.018 

0.475f 0.018 

1.218f 0.025 

0.994f 0.040 

0.919 f 0.033 

0.817f 0.047 

0.634 f 0.116g 

0.182 f 0.008 

0.203 f 0.013 

0.697f 0.011 

0.666f 0.015 

0.626 f 0.016 

0.643 f 0.024 

0.625 f 0.049g 
Neurobehavioral Studies 255 

TABLEI1 

Neurobehavioral Data for Rats in the 2-Year Gavage Study of Scopolamine Hydrobromide Trihydrate 

(continued) 

Female (continued) 

n 

Pawlick latency (seconds) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Startle response (milliseconds) 

Prestudy 

Day 1 

Day 90 

Day 180 

Day 270 

Day 360 

Day 720 

Passive avoidance (seconds) 

Day 90 

Day 91 

Day 180 

Day 270 

Day 360 

Day 720 

Vehicle Control 

10 

5.91 f 0.45 

5.34 f 0.42 

5.82 f 0.23 

6.06 f 0.16 

5.62 f 0.36 

4.97 f 0.40 

4.47 f 0.41h 

34.47 f 2.65 

36.72 f 4.02 

38.51 & 3.79 

27.09 f 3.64 

21.71 f 2.54 

31.61 f 2.45 

34.52 f 5.95g 

28.96 f 9.61 

496.5 f 82.0 

444.8 f 66.5 

579.0 f 63.4 

386.3 f 88.7 

279.8 f 119.4B 

10 

6.04 f 0.39 

5.30 f 0.33 

5.69 f 0.33 

5.72 f 0.49 

5.65 f 0.43 

4.96 f 0.30b 

5.30 f ].loB 

29.01 f 2.31 

33.65 f 6.52 

32.87 f 3.19 

24.42 f 2.51 

21.39 f 2.97 

28.82 f 2.71b 

29.78 f 3.02B 

18.13 f 3.88 

536.6 f 51.4 

499.6 f 57.8 

520.2 f 82.7 

352.3 f 105.2b 

38.2 f 11.4B 

10 

5.85 f 0.36 

4.64 f 0.28 

6.32 f 0.66 

5.69 f 0.28 

4.71 f 0.30b 

5.01 f 0.31b 

5.08 f 0.92B 

38.11 f 3.86 

42.27 f 5.15 

26.69 f 2.54* 

24.52 f 2.79 

18.75 f 2.12b 

33-01 f 4.20b 

29.26 f 49 

45.53 f 28.92 

396.5 f 80.6 

369.6 f 69.9 

419.4 f 93.8b 

525.9 f 74.9b 

255.7 f 116.9B 

25 mgntg 

10 

5.44 f 0.26 

6.82 f 1.15 

8.20 f 0.70** 

6.56 f 0.37b 

6.31 f 0.42b 

5.80 f 0.51' 

5.73 f 0.93h 

42.05 f 7.57 

36.66 f 4.36 

22.70 f 1.98** 

23.38 f 2.91b 

18.03 f 1.96b 

28.24 f 3.08f 

35.20 f 3.76' 

31.59 f 13.93 

465.7 f 48.3 

361.1 f 78.6b 

343.9 f 71.0b 

589.1 f 77.3' 

200.3 f 88.7' 

* Significantly different (P10.05)from the control group by Dunnett's test. 

** PSO.01 

a Mean f standard error. Statistical tests were performed on unrounded data. 

n=9 

n=5 * n= 1; no standard error calculated 

e n=6 

n=8 

g n=4 

n=3 
256 Scopolamine Hydrobromide Trihydrate, NTP TR 445 
257 

APPENDIX J 

CHEMICAL CHARACTERIZATION AND 

DOSE FORMULATION STUDIES 

PROCUREMENT AND CHARACTERIZATION OF sCOP0LA"E HYDROBROMIDETRENDRATE ....... 258 

PREPARATION AND ANALYSIS OF DOSEFORMULATIONS .............................. 259 

FIGURE J1 Infrared Absorption Spectrum of Scopolamine Hydrobromide Trihydrate ........ 261 

FIGURE 52 Nuclear Magnetic Resonance Spectrum of Scopolamine Hydrobromide Trihydrate ... 262 

TABLEJ1 Preparation and Storage of Dose Formulations in the Gavage Studies 

of Scopolamine Hydrobromide Trihydrate ............................. 263 

TABLE52 Results of Analysis of Dose Formulations Administered to Rats and Mice 

in the 16-Day Gavage Studies of Scopolamine Hydrobromide Trihydrate ......... 264 

TABLE53 Results of Analysis of Dose Formulations Administered to Rats and Mice 

in the 14-Week Gavage Studies of Scopolamine Hydrobromide Trihydrate ........ 265 

TABLE 54 Results of Analysis of Dose Formulations Administered to Rats and Mice 

in the 2-Year Gavage Studies of Scopolamine Hydrobromide Trihydrate ......... 267 

TABLE55 Results of Referee Analysis of Dose Formulations Administered 

to Rats and Mice in the 14-Week and 2-Year Gavage Studies 

of Scopolamine Hydrobromide Trihydrate .............................. 271 
258 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

CHEMICAL CHARACTERIZATION AND 

DOSE FORMULATION STUDIES 

PROCUREMENTANDCHARACTERIZATION 

OF SCOPOLAMINE HYDROBROMIDETRIHYDRATE 

Scopolamine hydrobromide trihydrate was obtained in two lots from Rebeco Chemicals, Inc. 

(New York, NY) (lot 14188) and from Henley and Company, Inc. (New York, NY) (lot 283). Lot 14188 

was used during the 16-day studies, and lot 283 was used during the 14-week and 2-year studies. Identity, 

purity, and stability were conducted by the analytical chemistry laboratory, Midwest Research Institute 

(Kansas City, MO). Reports on analyses performed in support of the scopolamine hydrobromide 

trihydrate studies are on file at the National Institute of Environmental Health Sciences. 

Both lots of the chemical, a white powder, were identified as scopolamine hydrobromide trihydrate by 

infrared, ultraviolet/visible, and nuclear magnetic resonance spectroscopy. All spectra were consistent 

with that expected for the structure, and the infrared and nuclear magnetic resonance spectra were 

consistent with the literature spectra (Sudtler Standard Spectra; Feeney, 1978) of scopolamine 

hydrobromide trihydrate (Figures J1 and 52). The observed melting point of 192"to 194" C and the 

specific optical activity ([a]?= -24.5 " f 0.1") for lot 14188 were consistent with the literature 

reference (Merck Index, 1983). 

The purity of both lots was determined by elemental analysis, Karl Fisher water analysis, functional group 

titration, thin-layer chromatography (TLC), and high-performance liquid chromatography (HPLC). Weight 

loss on drying was determined for lot 14188. Functional group titration was performed by dissolving 

samples of the test chemical in glacial acetic acid containing excess mercury (11) acetate and then titrating 

with 0.1 N perchloric acid in glacial acetic acid. The titrations were monitored potentiometrically using a 

combination pH/mV electrode filled with aqueous 4 M potassium chloride. TLC was performed on Silica 

Gel 60 F-254 plates with two solvent systems: 1) ch1oroform:acetone:diethylamine(50:40:10)and 

2)to1uene:ethyl acetate:diethylamine (70:20: 10)with caffeine as a reference standard. Plates were 

examined under ultraviolet light (254 nm) and with a spray of equal parts 0.3% hexachloroplatinic acid 

and 6% potassium iodide solution. HPLC was performed with a Waters pBondapak C,,column, a solvent 

system of 0.005M sodium heptanesulfonic acid in water:0.005 M sodium heptanesulfonic acid in 

methanol (62:38) with 10%phosphoric acid added to adjust the pH, a flow rate of 1.0 mL/minute, and 

ultraviolet detection at 229 nm. Lot 14188 and a solution of dried United States Pharmacopeia XX (USP) 

reference standard scopolamine hydrobromide were concomitantly analyzed with the same HPLC methods 

used for the purity analysis of lot 14188 but with acetanilide as a reference standard. Lot 283 was 

concomitantly analyzed with a solution of dried USP reference standard scopolamine hydrobromide and 

with lot 14188 with the same HPLC methods as the concomitant analysis of lot 141 88. 

For lot 14188, elemental analyses for carbon, hydrogen, nitrogen, and bromine were in agreement with 

the theoretical values for scopolamine hydrobromide trihydrate. Karl Fisher water analysis indicated 

10.8%f 0.3%water. Weight loss on drying indicated 10.8%f 0.02%water. Functional group 

titration indicated a purity of 101.9%f 0.1% scopolamine hydrobromide trihydrate, equivalent to 

89.3% f 0.1% anhydrous scopolamine hydrobromide. TLC withsystem 1 indicated a major spot, two 

trace impurities, and one slight trace impurity. TLC with system 2 indicated a major spot and one trace 

impurity. HPLC indicated a major peak and one impurity with a peak area of 0.1% relative to the major 

peak. Major peak comparisons of lot 14188 with a solution of dried USP reference standard scopolamine 

hydrobromide indicated that lot 14188 contained 88.8% f 0.2%scopolamine hydrobromide relative to the 

USP reference. Lot 14188 was determined to contain 89%scopolamine hydrobromide and 11 % water. 
Chemical Characterization and Dose Formulation 259 

The theoretical values for scopolamine hydrobromide trihydrate are 87.7 % scopolamine hydrobromide and 

12.3% water. 

For lot 283, elemental analyses for carbon, hydrogen, nitrogen, and bromine were in agreement with the 

theoretical values for scopolamine hydrobromide trihydrate. Karl Fisher water analysis indicated 

1 1.2% 0.2 % water.Functional group titration indicated a purity of 101.7% f 0.6% scopolamine 

hydrobromide trihydrate, equivalent to 89.2% f 0.5% anhydrousscopolamine hydrobromide. TLC 

indicated a major spot and one trace impurity by each system. HPLC indicated a major peak and one 

impurity with a peak area of 0.2%relative to the major peak. Major peak comparisons of lot 283 with a 

solution of dried USP reference standard scopolamine hydrobromide indicated that lot 283 contained 

89.2% If: 0.3% scopolamine hydrobromide relative to the USP reference. Major peak comparisons of 

lot 283 with lot 14188 indicated a purity of 100.0% A- 0.4% relative to lot 14188. Lot 283 was also 

determined to contain 89% scopolamine hydrobromide and 11 % water. 

Reanalyses to determine the purity of lot 283 were conducted by the analytical chemistry laboratory using 

functional group titration and HPLC. Functional group titration was performed as previously described 

except that the combination pH/mV electrode was filled with aqueous 3 M potassium chloride. Frozen 

reference samples of lots 14188 and 283 were concomitantly analyzed using the same HPLC methods as in 

the original purity analysis of lot 283 and using those same HPLC methods but with acetanilide as an 

internal standard and a solvent ratio of 65: 35. Functional group titration indicated a purity of 

102.0% f 0.6%. Major peak comparisons lot 283 with lot 14188 by the first HPLC system indicated no 

additional impurity peaks. Major peak comparisons of lot 283 with lot 14188 by the second HPLC system 

indicated a purity of 100.3 % f 0.5% relative to lot 14188. 

Stability studies of lot 14188 of the bulk chemical were performed by the analytical chemistry laboratory. 

Analyses were performed using the same HPLC protocol described for the purity analysis but with 

acetanilide as an internal standard. These studies indicated that bulk scopolamine hydrobromide trihydrate 

was stable when stored for up to 2 weeks in sealed containers, protected from light, under a nitrogen 

headspace, and at temperatures up to 25" C. To ensure stability, the bulk chemical was stored in amber 

glass jars at approximately 25°C under a nitrogen headspace. Stability of the bulk chemical was 

monitored at the beginning of the 16-day studies and at the beginning and end of the 14-week studies using 

HPLC. For the 2-year studies, stability of the bulk chemical was monitored at the beginning and end of 

the studies and every 4 months during the studies using HPLC and potentiometric titration. No 

degradation of the bulk chemical was detected. 

mPARATION AND ANALYSIS OF DOSEFORMULATIONS 

Dose formulations for the 16-day and 14-week studies were prepared every two weeks. For the 2-year rat 

study, dose formulations were prepared weekly during the first four months of the study and every two 

weeks for the remainder of the study. Dose formulations for the 2-year mouse study were prepared every 

two weeks throughout the study. Dose formulations were prepared by mixing the appropriate quantities of 

scopolamine hydrobromide trihydrate with deionized water to give the required concentrations (Table J1) 

and were stored at room temperature in amber glass bottles capped with Teflon@-lined lids in the dark for 

up to 3 weeks. 

Stability studies of the 11 and 0.2 mg/mL dose formulations were conducted by the analytical chemistry 

laboratory. For the 11 mg/mL formulation, samples were prepared by dissolving scopolamine 

hydrobromide trihydrate in deionized water and diluting to volume. Aliquots (5 mL) were mixed with 

internal standard solution (0.4 mg/mL acetophenone in acetonitrile) and were further diluted with 

0.007 M aqueous sodium heptanesulfonic acid. The samples were then analyzed by HPLC using a 
260 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

Waters C,, Nova Pak column, with a mobile phase of water:acetonitrile (70:30), at a flow rate of 

1.0 mWminute, and with ultraviolet detection at 229 nm. For the 0.2 mg/mL formulation, samples were 

prepared by dissolving scopolamine hydrobromide trihydrate in deionized water and diluting to volume. 

Aliquots (7mL) were mixed with internal standard solution (0.04 mg/mL propiophenone in acetonitrile) 

and were analyzed by HPLC with the same system used to analyze the 11 mg/mL formulation but with a 

mobile phase ratio of 75:25. The stability of the dose formulations was confirmed for at least 3 weeks at 

room temperature when stored in the dark or for at least 3 hours at room temperature and open to air and 

light. 

Periodic analyses of the dose formulations of scopolamine hydrobromide trihydrate were conducted at the 

study laboratory using ultraviolet/visible spectrometry for the 16-day and 14-week studies and by HPLC 

for the 2-year studies. During the 16-day studies, dose formulations were analyzed at the beginning of the 

studies (Table 52). During the 14-week and 2-year studies, dose formulations were analyzed every 6 to 

8 weeks (Tables 53 and 54). During the 16-day studies, 70% (7110) of the dose formulations were within 

10% of the target concentration with no value exceeding 16% of the target concentration. All of the dose 

formulations analyzed during the 14-week studies were within 10% of the target concentration. During 

the 2-year studies, 92% (72/78) of the dose formulations were within 10% of the target concentration with 

no value differing more than 20% from the target concentration. For the 14-week and 2-year studies, 

results of periodic referee analyses performed by the analytical chemistry laboratory indicated good 

agreement with the results of the study laboratory (Table J5). 
Chemical Characterization and Dose Formulation 261 

0 

v) 

v) 

3 N 

0 

N 

z z 

N 

(D

N 

w 

0

z

- W 

0 U 

W 

U 

W

U 

-

z 

I 

z

0

F

<

a 

t3 5 B 

s 

x 0

0 

FIGURE J1 

Infrared Absorption Spectrum of Scopolamine Hydrobromide Trihydrate 
i; e

h 

P 

Li 

m 

c1 

ai

L ll'L{ 

I I6'0 

t oo'C 
Chemical Characterization and Dose Formulation 263 

of Scopolamine Hydrobromide Trihydrate 

Preparation and Storage of Dose Formulations in the Gavage Studies 

TABLEJ1 

16-Day Studies 14-Week Studies 2-Year Studies 

desired concentrations 

deionized water and diluted to the 

third of the required volume of 

was dissolved in approximately one 

Scopolamine hydrobromide trihydrate 

Preparation 

Same as 16-day studies Same as 16-day studies 

14188 

Chemical Lot Number 

283 283 

3 weeks 

Maximum Storage Time 

3 weeks 3 weeks 

temperature and protected from light 

Stored in sealed containers at room 

Storage Conditions 

and protected from light 

Teflon"-lined lids at room temperature 

Stored in amber glass bottles with Same as 14-week studies 

(Birmingham, AL) 

Southern Research Institute 

Study Laboratory 

(Birmingham, AL) 

Southern Research Institute 

(Columbus, OH) 

Battelle Columbus Laboratories 

(Kansas City, MO) 

Midwest Research Institute 

Referee Laboratory 

(Kansas City, MO) 

Midwest Research Institute 

(Kansas City, MO) 

Midwest Research Institute 
264 Scopolamine Hydrobromide Trihydrate, NTP TFt 445 

TABLE52 

Results of Analysis of Dose Formulations Administered to Rats and Mice in the 16-Day Gavage Studies 

of Scopolamine Hydrobromide Trihydrate 

Determined Target Difference 

Date Prepared Analyzed Date ConcentrationConcentration from Target 

(mg/mL) (mg/mL)a (“/.I 

Rats 

27 June 1985 27 June 1985 15 15.2 +1 

30 30.1 0 

60 59.7 -1 

120 114.4 -5 

240 209.2 -13 

9 July 1985b 15 15.0 0 

30 29.7 -1 

60 60.0 0 

120 114.4 -5 

1 July 1985‘ 1 July 1985 240 210.6 -12 

9 July 1985b 240 214.0 -11 

Mice 

2 July 1985 3 July 1985 15 13.9 -7 

25 22.9 -8 

45 41.4 -8 

90 78.2 -13 

180 150.8 -16 

12 July 19Ub 15 14.0 -7 

25 23.O -8 

45 41.3 -8 

4 July 1985‘ 5 July 1985 90 79Xd -11 

180 153.0d -15 

12 July 1985 90 88.0 -2 

180 166.7 -7 

12 July 1985b 90 88.6 -2 

180 166.6 -7 

a Results of duplicate analyses. Dosing volume = 0.5 mLl100 g (rats); 15 mglmL = 75 mglkg, 30 mglmL = 150 pglkg, 

60 mglmL = 300 mglkg, 120 mglmL = 600mglkg, 240 mglmL = 1,200 mg/kg. Dosing volume = 0.1 mLl10 g (mice); 

15 mglmL = 150 mglkg, 25 mglmL = 250 mglkg, 45 mglmL = q50 mglkg, 90 mglmL = 900 mglkg, 180 mglmL = 1,800 mglkg. 

Animal room sample ‘ Results of remix 

Trial for new dose formulation procedure; formulation not used for “sing 
Chemical Characterization and Dose Formulation 265 

TABLE53 

Results of Analysis of Dose Formulations Administered to Rats and Mice in the 14-Week Gavage Studies 

of Scopolamine Hydrobromide Trihydrate 

Prepared DateDate Analyzed 

Rats 

7 March 1986 10 March 1986 

28 March 1986b 

18 April 1986 18 April 1986 

9 May 1986b 

30 May 1986 2 June 1986 

19 June 1986b 

Mice 

7 March 1986 10 March 1986 

28 March 1986b 

Target Concentration 

(mg/mL) 

3.0 

9.0 

27.0 

80.0 

240.0 

3.0 

9.0 

27.0 

80.0 

240.0 

3.0 

9.0 

27.0 

80.0 

240.0 

3.0 

9.0 

27.0 

80.0 

240.0 

3.0 

9.0 

27.0 

80.0 

240.0 

3.0 

9.0 

27.0 

80.0 

240.0 

1.5 

4.5 

13.5 

40.0 

120.0 

1.5 

4.5 

13.5 

40.0 

120.0 

Determined 

Concentration 

(mg/mL)a 

2.92 

8.88 

26.8 

80.2 

241.7 

3.00 

9.06 

26.9 

80.2 

238 

2.96 

8.70 

26.9 

79.0 

239 

2.91 

8.78 

27.1 

79.3 

240 

3.04 

8.98 

26.2 

79.0 

237 

2.95 

8.80 

26.0 

77.6 

239 

1.46 

4.45 

13.4 

39.6 

119.3 

1.54 

4.70 

13.5 

39.8 

119 

Difference 

from Target 

("/.I 

-3 

-1 

-1 

0 

+1 

0 

+1 

0 

0 

-1 

-1 

-3 

0 

-1 

0 

-3 

-2 

0 

-1 

0 

+I 

0 

-3 

-1 

-1 

-2 

-2 

-4 

-3 

0 

-3 

0 

-1 

-1 

-1 

+3 

+4 

0 

-1 

-1 
266 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLE53 

Results of Analysis of Dose Formulations Administered to Rats and Mice in the 14-Week Gavage Studies 

of Scopolamine Hydrobromide Trihydrate (continued) 

Determined Target Difference 

Prepared DateAnalyzedDate Concentration Concentration from Target

(mg/mL) (mg/mL) (%I 

Mice (continued) 

18 April 1986 18 April 1986 1.5 1.44 -4 

4.5 4.32 -4 

13.5 12.8 -5 

40.0 39.0 -3 

120.0 118 -2 

9 May 1986b 1.5 1.51 +1 

4.5 4.38 -3 

13.5 12.9 -4 

40.0 38.7 -3 

120.0 118 -2 

13 June 1986 16June 1986 1.5 1.54 +3 

4.5 4.52 +1 

13.5 13.5 0 

40.0 40.O 0 

120.0 122 +2 

27June1986b 1.5 1.54 +3 

4.5 4.48 0 

13.5 13.2 -2 

40.0 39.6 -1 

120.0 122 +2 

a Results of duplicateanalyses.Dosingvolume = 0.5 mLll00 g (rats); 3.0 mglmL = 15mglkg, 9.0 mglmL = 45 mglkg, 

27.0 mglmL = 135 mglkg, 80 rnglmL = 400mglkg,240mglmL = 1,200 mglkg. Dosing volume = 0.1 mLll0 g (mice); 

1.5 mglmL = 15 mglkg. 4.5 mglmL = 45 mglkg, 13.5 mglmL = 135 mglkg, 40 mglmL = 400mglkg, 120 rnglmL = 1.200 mglkg. 

Animal room sample 
Chemical Characterization and Dose Formulation 267 

TABLE54 

Results of Analysis of Dose Formulations Administered to Rats and Mice in the 2-YearGavage Studies 

of Scopolamine Hydrobromide Trihydrate 

Prepared DateDate Analyzed 

Rats 

11 October 1988 

21 November 1988 

17 January 1989 

14 March 1989 

9 May 1989 

6 July 1989 

29 August 1989 

24 October 1989 

19 December 1989 

14 October 1988 

28 October 1988b 

21 November 1988 

18-19 January 1989 

15 March 1989 

28-29 March 1989b 

1 1May 1989 

7-8 July 1989 

31 August-1 September 1989 

13-14 September 1989b 

27 October 1989 

20 December 1989 

Target Concentration 

(mgld) 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

0.2 

1.o 

5.0 

Determined 

Concentration 

(mgh”, 

0.203 

0.990 

5.053 

0.195 

0.995 

4.853 

0,203 

0.956 

4.928 

0.202 

1.025 

4.958 

0.201 

0.962 

4.839 

0.205 

1.081 

5.047 

0.201 

0.936 

4.895 

0.239 

1.201 

5.617 

0.210 

1.024 

5.328 

0.209 

1.101 

5.035 

0.210 

1.039 

5.115 

0.207 

1.050 

4.864 

Difference 

from Target 

6) 

+2 

-1 

+1 

-3 

-1 

-3 

+2 

I-4 

-1 

+1 

+3 

-1 

+1 

-4 

-3 

+3 

+8 

+1 

+1 

-6 

-2 

+20 

+20 

+ 12 

+5 

+2 

+7 

+5 

+10 

+1 

+5 

+4 

+2 

+4 

+5 

-3 
268 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLE54 

Results of Analysis of Dose Formulations Administered to Rats and Mice in the 2-YearGavage Studies 

of Scopolamine Hydrobromide Trihydrate(continued) 

Target 

Date Prepared Date Analyzed Concentration 

(mglmL) 

Rats (continued) 

13 February 199016 FebruaIy 1990 0.2 

1 .o 

5.O 

6 March 1990b 0.2 

1 .o 

5.0 

16 April 1990 18 April 1990 0.2 

1 .o 

5.0 

2 May 1990b 0.2 

11 June 1990 12June 1990 0.2 

1 .o 

5.0 

6 August 1990 8 August 1990 0.2 

1 .o 

5.0 

31 August 1990b 0.2 

1 .o 

5.O 

Mice 

13 September 1988 15 September 1988 0.1 

0.5 

2.5 

30 September 198tlb 0.1 

0.5 

2.5 

21 November 198821 November 1988 0.1 

0.5 

2.5 

17 January 198918-19 January 1989 0.1 

0.5 

2.5 

Determined 

Concentration 

(mglmL) 

0.204 

0.998 

4.994 

0.159 

0.999 

5.006 

0.205 

0.999 

5.061 

0.179 

0.205 

1.062 

5.250 

0.204 

0.995 

5.047 

0.116 

1.039 

4.982 

0.097 

0.498 

2.49 

0.094 

0.494 

2.46 

0.100 

0.507 

2.534 

0.100 

0.489 

2.435 

Difference 

from Target 

(%I 

+2 

0 

0 

-21 

0 

0 

+3 

0 

+1 

-11 

+3 

+6 

+5 

+2 

-1 

+1 

-42 

+4 

0 

-3 

0 

0 

-6 

-1 

-2 

0 

+1 

+1 

0 

-2 

-3 
Chemical Characterization and Dose Formulation 269 

TABLE54 

Results of Analysis of Dose Formulations Administered to Rats and Mice in the 2-YearGavage Studies 

of Scopolamine Hydrobromide Trihydrate (continued) 

Determined Target 

Date Prepared Analyzed Date Concentration Concentration 

(w/mL) 

Mice (continued) 

14 March 1989 15 March 1989 0.1 

0.5 

2.5 

28-29 March 1989b 0.1 

0.5 

2.5 

9 May 1989 11May 1989 0.1 

0.5 

2.5 

6 July 1989 7-8 July 1989 0.1 

0.5 

2.5 

29 August 1989 31August-1September1989 0.1 

0.5 

2.5 

13-14 September 198gb 0.1 

0.5 

2.5 

24 October 1989 27 October 1989 0.1 

OS 

2.5 

19 December 1989 20-21 December 1989 0.1 

0.5 

2.5 

13February 1990 16 February 1990 0.1 

0.5 

2.5 

6 March 1990b 0.1 

0.5 

2.5 

16 April 1990 18 April 1990 0.1 

0.5 

2.5 

2 May 1990b 0.1 

0.5 

0.095 

0.499 

2.522 

0.106 

0.494 

2.511 

0.093 

0.508 

2.518 

0.1 14 

0.575 

2.907 

0.102 

0.539 

2.563 

0.112 

0.511 

2.582 

0.104 

0.523 

2.554 

0.103 

0.498 

2.480 

0.101 

0.500 

2.530 

0.085 

0.442 

2.41 1 

0.105 

0.516 

2.480 

0.058 

0.470 

Difference 

from Target 

("/.I 

-5 

0 

+1 

+6 

-1 

0 

-7 

+2 

+1 

+14 

+15 

+16 

+2 

+8 

+3 

+12 

+2 

+3 

+4 

+5 

+2 

+3 

0 

-1 

+1 

0 

+1 

-15 

-12 

-4 

+5 

+3 

-1 

-42 

-6 
270 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLE54 

Results of Analysis of Dose Formulations Administeredto Rats and Mice in the 2-Year Gavage Studies 

of Scopolamine Hydrobromide Trihydrate(continued) 

Difference Determined Target 

Date hepared Date Analyzed Target ConcentrationConcentration from 

(mg/mL) (mg/mL) 

Mice (continued) 

11 June 1990 12 June 1990 0.1 0.106 +6 

0.5 0.511 +2 

2.5 2.574 +3 

6 August 1990 , 8 August 1990 0.1 0.101 +1 

0.5 0.507 +1 

2.5 2.492 0 

31 August 1990b 0.1 0.040 -60 

0.5 0.450 -10 

2.5 2.525 +1 

a Results of duplicate analyses. Dosing volume = 0.5 mLl100 g (rats); 0.2 mglmL = 1.0 mglkg, 1.0 mglmL = 5 mglkg, 

5.0 mglmL = 25 mglkg. Dosing volume = 1 mLl100 g (mice); 0.1 mglmL = 1.0 mglkg, 0.5 mglmL = 5 mglkg, 

2.5 mglmL = 25 mglkg. 

Animal room sample 
Chemical Characterization and Dose Formulation 271 

TABLEJ5 

Results of Referee Analysis of Dose Formulations Administered to Rats and Mice 

in the 14-Week and 2-Year Gavage Studies of Scopolamine Hydrobromide Trihydrate 

DeterminedConcentration(rndmL1 

Target Concentration Study Referee 

DatePrepared (mg/d) Laboratorya Laboratoryb 

14-Week Studies(Southern ResearchInstitute) 

Rats 

7 March 1986 3.0 

Mice 

13 June 1986 120.0 122 119.00 f 1.00 

2-YearStudies (J3attelleColumbus Laboratories) 

Rats 

11 October 1988 0.2 0.203 0.201 f 0.001 

14 March 1989 1.o 0.962 1.010 f 0.01 

13 February 1990 5.0 4.994 4.92 f 0.04 

Mice 

13 September 1988 2.5 2.49 2.48 f 0.02 

29 August 1989 0.5 0.539 0.508 f 0.01 

a Results of duplicate analyses 

Results of triplicate analyses (mean f standard error) 
272 Scopolamine Hydrobromide Trihydrate, NTP TR 445 
273 

APPENDIX K 

INGREDIENTS, NUTRIENT COMPOSITION, 

AND CONTAMINANT LEVELS 

IN NIH-07 RAT AND MOUSE RATION 

TABLEK1 Ingredients of NJH-07 Rat and Mouse Ration ........................... 274 

TABLEK2 Vitamins and Minerals in NIH-07 Rat and Mouse Ration................... 274 

TABLEK3 Nutrient Compositionof NM-07 Rat and Mouse Ration .................... 275 

TABLEK4 Contaminant Levels inNIH-07 Rat and Mouse Ration ..................... 276 
274 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

TABLEK1 

Ingredients of NIH-07Rat 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 

Alfalfameal (dehydrated, 17% protein) 4.00 

Corn gluten meal (60% protein) 3.00 

Soy oil 2.50 

Dried brewer's yeast 2.00 

Dry molasses 1SO 

Dicalcium phosphate 1.25 

Ground limestone 0.50 

Salt 0.50 

Premixes (vitamin and mineral) 0.25 

a NCI, 1976;NM, 1978 

Ingredients were ground to pass through a U.S. Standard Screen No. 16 before being mixed. 

TABLEK2 

Vitamins and Minerals inNM-07 Rat and Mouse Rationa 

Amount Source 

Vitamins 

A 5,500.Ooo IU Stabilized vitaminA palmitate or acetate 

D3 4,600,OooIU D-activated animal sterol

2.8 g Menadione K 3

d-a-Tocopheryl acetate 20,Ooo IU 

Choline 560.0 g Choline chloride 

Folic acid 2.2 g 

Niacin 30.0g 

d-Pantothenic acid 18.0g &Calcium pantothenate 

Riboflavin 3.4 g 

Thiamine 10.0 g Thiamine mononitrate 

*12 4.OOO Pg

Qridoxine 1.7g Pyridoxine hydrochloride 

Biotin 140.0 mg d-Biotin 

Minerals 

Iron 120.0 g Iron sulfate 

Manganese 60.0g Manganous oxide 

Zinc 16.0g Zinc oxide 

Copper 4.0 g Ccpper sulfate 

Iodine 1.4g Calciumiodate ' 

Cobalt 0.4g Cobalt carbonate 

a Per ton (2,OOOIb)offinished product 
Feed Analyses 275 

TABLEK3 

Nutrient Compositionof NIH-07 Rat and Mouse Ration 

Nutrient 

Protein (% by weight) 

Crude fat (Xby weight) 

Crude fiber (% by weight) 

Ash (% by weight) 

AminoAcids (% of total diet) 

Arginine 

Cystine 

Glycine 

Histidine 

Isoleucine 

Leucine 

Lysine 

Methionine 

Phenylalanine 

Threonine 

Tryptophan 

Tyrosine 

Valine 

Essential Fatty Acids (% of total diet) 

Linoleic 

Linolenic 

Vitamins 

Vitamin A (IUlkg) 

Vitamin D (IU/kg) 

a-Tocopherol (ppm) 

Thiamine (ppm) 

Riboflavin (ppm) 

Niacin (ppm) 

Pantothenic acid (ppm) 

Pyridoxine (ppm) 

Folic acid (ppm) 

Biotin (ppm) 

Vitamin B12 (ppb) 

Choline (ppm) 

Minerals 

Calcium (%) 

Phosphorus (%) 

Potassium (%) 

Chloride (%) 

Sodium (%) 

Magnesium (%) 

Sulfur (X) 

Iron (ppm) 

Manganese (ppm) 

Zinc (ppm) 

Copper (ppm) 

Iodine (ppm) 

Chromium (ppm) 

Cobalt (ppm) 

Mean f Standard 

Deviation 

23.17 f 0.71 

5.27 f 0.21 

3.55 f 0.41 

6.47f 0.24 

1.287f 0.084 

0.306 f 0.075 

1.160 f 0.050 

0.580 f 0.024 

0,917f 0.034 

1.972 f 0.052 

1.273 f 0.051 

0.437 f 0.115 

0.994 f 0.125 

0.896 f 0.055 

0.233f 0.160 

0.677 f 0.105 

1.089 f 0.057 

2.389f 0.233 

0.277 f 0.036 

6,798f 1,866 

4,450 f 1,382 

36.92f 2.20 

18.60 f 2.19 

7.92f 0.93 

100.95 f 25.92 

30.30f 3.60 

9.25 f 2.62 

2.51 f 0.64 

0.267 f 0.049 

40.14 f 20.04 

3,068 f 314 

1.21 f 0.11 

0.94f 0.03 

0.887 f 0.067 

0.526 f 0.092 

0.315 f 0.034 

0.168 f 0.008 

0.274 f 0.063 

356.2f 90.0 

92.24 f 5.35 

58.14 f 9.91 

11.50 f 2.40 

3.70f 1.14 

1.71 f 0.45 

0.797 f 0.23 

Range 

21.80 -24.30 

4.60-5.70 

2.60-4.30 

6.11 -7.30 

1.100- 1.390 

1.181-0.400 

1.060 - 1.220 

0.531 -0.608 

0.867-0.965 

1.850 -2.040 

1.200- 1.370 

0,306-0.699 

0.665 - 1.110 

0.824-0.985 

0,107-0.671 

0.564 -0.794 

0.962 - 1.170 

1.830-2.570 

0.210-0.320 

4,180-12,140 

3,OOO -6,300 

22.5 -48.9 

15.0-28.0 

6.10-9.00 

65.0- 150.0 

23.0-34.6 

5.60-14.0 

1.80-3.70 

0.19 -0.35 

10.6-65.0 

2,400- 3,430 

1.00-1.54 

0.85 - 1.00 

0.772-0.971 

0.380-0.635 

0.258 -0.370 

0.151 -0.180 

0.208 -0.420 

255.0 -523.0 

81.70-99.40 

46.10 -81.60 

8.090- 15.39 

1.52-5.83 

0.85 -2.09 

0.490-1.150 

Number of Samples 

37 

37 

37 

37 

10 

10 

10 

10 

10 

10 

10 

10 

10 

10 

10 

10 

10 

9 

9 

37 

4 

9 

37 

10 

9 

10 

10 

10 

10 

10 

9 

37 

37 

8 

8 

10 

10 

10 

10 

10 

10 

10 

10 

9 

6 
276 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

TABLEK4 

Contaminant Levels in NJH-07Rat and Mouse Rationa 

Mean f Standard 

Deviationb Range Number of Samples 

Contaminants 

Arsenic (ppm) 0.29f 0.19 0.06-0.70 37 

Cadmium (ppm) 0.09 f 0.05 0.05-0.2 37 

Lead (ppm) 0.27f 0.20 0.10- 1.00 37 

Mercury (ppm) 0.03 f 0.02 0.02 -0.11 37 

Selenium (ppm) 0.38f 0.22 0.05 - 1.27 37 

Aflatoxins (ppb)' <5.0 37 

Nitrate nitrogen (p~rn)~ 14.82f 4.80 5.70-24.0 37 

Nitrite nitrogen (ppm)d 0.21 f 0.17 0.10-0.07 37 

BHA (PP~)~ 1.78 f 1.62 1.00- 10.00 37 

BHT (pp& 1.54f 1.35 1 .OO -8.00 37 

Aerobic plate count (CFU/g) 43,275 f 27,682 4,100- 12O.OOO 37 

Coliform (MPNlg) 4.05 f 4.36 3.00-23.00 37 

Escherichia coli (MPN/g) 3.oo <3.00 38 

Salmonella (MPN/g) Negative 25 

Total nitrosoamines (ppb)f 7.57f 2.60 3.60- 16.50 37 

N-Nitrosodimethylamine (p bp 5.67 f 2.26 2.60- 13.00 37 P N-Nitrosopyrrolidine (ppb) 1.92f 1.06 1.00 -4.30 37 

Pesticides (ppm) 

(Y-BHC <0.01 31 

0-BHC <0.02 31 

?-BHC <0.01 31 

&BHC <0.01 31 

Heptachlor <0.01 31 

Aldrin <0.01 31 

Heptachlor epoxide <0.01 31 

DDE <0.01 31 

DDD <0.01 31 

DDT <0.01 31 

HCB <0.01 31 

Mirex <0.01 31 

Methoxychlor <0.05 31 

Dieldrin <0.01 31 

Endrin <0.01 31 

Telodrin <0.01 31 

Chlordane <0.05 31 

Toxaphene <0.1 31 

Estimated PCBs <0.2 31 

Ronnel <0.01 31 

Ethion <0.02 31 

Trithion <0.05 31 

Diazinon <0.1 31 

Methyl parathion <0.02 31 

Ethyl parathion <0.02 31 

Malathion 0.24f 0.24 0.05 -1.00 37 

Endosulfan I <0.01 31 

Endosulfan I1 <0.01 31 

Endosulfan sulfate <0.03 31 

~ ~~ ~ ~~ ~ ~~~ ~ ~ 

a CFU = colony-forming units, MPN = most probable number, BHC = hexachlorocyclohexane or benzene hexachloride 

For values less than the limit of detection, the detection limit is given as the mean. 

' No aflatoxin measurement was recorded for the lot milled on 2 October 1989. 

Sources of contamination: alfalfa, grains, and fish meal. 

e Sources of contamination: soy oil and fish meal. 

All values were corrected for percent recovery. 
277 

APPENDIX L 

SENTINEL ANIMAL PROGRAM 

METHODS . ........................................................ 278 

TABLEL1Murine Virus AntibodyDeterminations for RatsandMice 

in the 14-Week and 2-Year Gavage Studies 

of ScopolamineHydrobromideTrihydrate . . . .. . ....... . ....... . ..... . . 280 
278 Scopolamine Hydrobromide Trihydrate, NTP TR 445 

SENTINEL ANIMAL PROGRAM 

METHODS 

Rodents used in the Carcinogenesis Program of the National Toxicology Program are produced in optimally 

clean facilities to eliminate potential pathogens that may affect study results. The Sentinel Animal Program is 

part of the periodic monitoring of animal health that occurs during the toxicologic evaluation of chemical 

compounds. Under this program, the disease state of the rodents is monitored via serology on sera from extra 

(sentinel) animals in the study rooms. These animals and the study animals are 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, allowed to clot, and the serum separated. The samples were processed 

appropriately and sent to Microbiological Associates, Inc. (Bethesda, MD), for determination of antibody titers. 

The laboratory serology methods and viral agents for which testing was performed are tabulated below; the 

times at which blood was collected during the studies are also listed. 

Method and Test Time of Analvsis 

RATS 

14-WeekStudy 

ELISA 

Mycoplasma arthntidis Study termination 

Mycoplasma pulmonis Study termination 

PVM (pneumonia virus of mice) Study termination 

RCVISDA Study termination 

(rat coronavirus/sialodacryoadenitisvirus) 

Sendai Study termination 

Hemagglutination Inhibition 

H-1 (Toolan’s H-1 virus) Study termination 

KRV (Kilham rat virus) Study termination 

2-Year Study 

ELISA 

M. arthritidis 24 months 

M. pulmonis 24 months 

PVM Quarantine, 6, 12, 18, and 24months 

RCVISDA Quarantine, 6, 12, 18, and 24months 

Sendai Quarantine, 6, 12, 18, and 24months 

Immunofluorescence Assay 

RCV/SDA ., 24 months 

Hemagglutination Inhibition 

Quarantine, 6, 12, 18, and 24months 

KRV Quarantine, 6, 12, 18, and 24months 

H-1 
Sentinel Animal Program 279 

MICE 

14-Week Study 

Complement Fixation 

LCM (lymphocytic choriomeningitis virus) 

ELISA 

Ectromelia virus 

GDVII (mouse encephalomyelitis virus) 

Mouse adenoma virus 

MHV (mouse hepatitis virus) 

M.arthritidis 

M.pulmonis 

PVM 

Reovirus 3 

Sendai 

Immunofluorescence Assay 

EDIM (epizootic diarrhea of infant mice) 

Hemagglutination Inhibition 

K (papovavirus) 

MVM (minute virus of mice) 

Polyoma virus 

2-Year Study 

ELISA 

Ectromelia virus 

EDIM 

GDVII 

LCM 

MVM 

Mouse adenoma virus 

MHV 

M.arthritidis 

M.pulmonis 

PVM 

Reovirus 3 

Sendai 

Immunofluorescence Assay 

EDIM 

GDVII 

LCM 

MVM 

Mouse adenoma virus 

MHV 

Reovirus 3 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Study termination 

Quarantine, 6, 12,18, and 24 months 

24 months 

Quarantine, 6, 12,18, and 24 months 

12, 18, and 24 months 

Quarantine, 6, and 12 months 

Quarantine, 6, 12, 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 

Quarantine, 6, 12,18, and 24 months 

24 months 

Quarantine and 6 months 

18 months 

18 and 24 months 

6 and 12 months 

24 months 
280 Scopolamine Hydrobromide Trihydrate,NTP TR 445 

MICE(continued) 

2-Year Study (continued) 

Hemagglutination Inhibition 

K Quarantine, 6, 12, 18, and 24 months

MVM 24 months 

Polyoma virus Quarantine, 6, 12, 18, and 24 months

Results of serology tests are presented in Table L1. 

TABLEL1 

Murine Virus Antibody Determinations for Rats and Mice in the 14-Week and 2-YearGavage Studies 

of Scopolamine Hydrobromide Trihydrate 

Interval Incidence of Antibody Positive Serologic 

in Sentinel Animals Reaction for 

14-Week Studies 

Rats 

Study termination 0110 None positive 

Mice 

Study termination 0110 None positive 

2-Year Studies 

Rats 

Quarantine 0110 None positive 

6 Months 0110 None positive 

12 Months 0110 None positive 

18 Months 0110 None positive 

24 Months 3110 M. arthritidi? 

Mice 

Quarantine 0110 None positive 

6 Months 9/10 MHVb 

12 Months 6/10 MHV 

18 Months 6110 MHV 

24 Months 10110 MHV 

1110 Reovirus 3 

1/10 M. arthritidis 

a Further evaluation of samples positive for M. arthritidis by immunoblot and Western blot procedures indicated that the positive titersmay 

be due to cross reaction with antibiotics of nonpathogenic Mycoplasma or other agents. Only sporadic samples were positive and there 

were no clinical findings or histopathic changes of M.athritidis infection in mice with positive titers. Accordingly, sporadic 

M.arthriridis-positive titers were considered to be false positive. 

MHV was positive by ELISA in 8 of 10 animals and positive by immunofluorescence assay in 9 of 10 animals 
DEPARTMENT OF 

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National Toxicology Program 

Central Data Management 

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March 1997