Allyl Acetate, Allyl Alcohol, and Acrolein

178 Pages·2006·16.13 MB·English

National Toxicology Program
Toxicity Report Series
Number 48
NTP Technical Report

on the Comparative Toxicity Studies of

Allyl Acetate, Allyl Alcohol, and Acrolein

(CAS Nos. 591-87-7, 107-18-6, and 107-02-8)
Administered by Gavage

to F344/N Rats and B6C3F Mice

1
July 2006

National Institutes of Health

Public Health Service

U.S. Department of Health and Human Services
FOREWORD
The National Toxicology Program (NTP) is an interagency program within the Public Health Service (PHS) of the
Department of Health and Human Services (HHS) and is headquartered at the National Institute of Environmental
Health Sciences of the National Institutes of Health (NIEHS/NIH). Three agencies contribute resources to the
program: NIEHS/NIH, the National Institute for Occupational Safety and Health of the Centers for Disease Control
and Prevention (NIOSH/CDC), and the National Center for Toxicological Research of the Food and Drug
Administration (NCTR/FDA). Established in 1978, the NTP is charged with coordinating toxicological testing
activities, strengthening the science base in toxicology, developing and validating improved testing methods, and
providing information about potentially toxic substances to health regulatory and research agencies, scientific and
medical communities, and the public.
The Toxicity Study Report series began in 1991. The studies described in the Toxicity Study Report series are
designed and conducted to characterize and evaluate the toxicologic potential of selected substances in laboratory
animals (usually two species, rats and mice). Substances selected for NTP toxicity studies are chosen primarily on
the basis of human exposure, level of production, and chemical structure. The interpretive conclusions presented in
the Toxicity Study Reports are based only on the results of these NTP studies. Extrapolation of these results to
other species, including characterization of hazards and risks to humans, requires analyses beyond the intent of
these reports. Selection per se is not an indicator of a substance’s toxic potential.
The NTP conducts its studies in compliance with its laboratory health and safety guidelines and FDA Good
Laboratory Practice Regulations and must meet or exceed all applicable federal, state, and local health and safety
regulations. Animal care and use are in accordance with the Public Health Service Policy on Humane Care and Use
of Animals. Studies are subjected to retrospective quality assurance audits before being presented for public
review.
NTP Toxicity Study Reports are indexed in the NIH/NLM PubMed database and are available free of charge
electronically on the NTP website (http://ntp.niehs.nih.gov) or in hardcopy upon request from the NTP Central Data
Management group at [email protected] or [phone].
National Toxicology Program
Toxicity Report Series
Number 48
NTP Technical Report

on the Comparative Toxicity Studies of

Allyl Acetate, Allyl Alcohol, and Acrolein
(CAS Nos. 591-87-7,107-18-6, and 107-02-8)
Administered by Gavage

to F344/N Rats and B6C3F Mice

1
Rick D. Irwin, Ph.D., Study Scientist
National Toxicology Program

P.O. Box 12233

Research Triangle Park, NC 27709

NIH Publication No. 06-4413
National Institutes of Health

Public Health Service

U.S. Department of Health and Human Services
2
CONTRIBUTORS

National Toxicology Program NTP Pathology Working Group
Evaluated and interpreted results and reportedf indings Evaluated slides and prepared pathology report ofallyl acetate
(January 8, 1997)
R.D. Irwin, Ph.D., Study Scientist
J.R. Bucher, Ph.D. J.C. Seely, D.V.M., Chairperson
L.T. Burka, Ph.D. PATHCO, Inc.
S. Botts, M.S., D.V.M., Ph.D.
R.E. Chapin, Ph.D.
Experimental Pathology Laboratories, Inc.
R.S. Chhabra, Ph.D.
R.A. Herbert, D.V.M., Ph.D.
J. Mahler, D.V.M.
National Toxicology Program
C.S. Smith, Ph.D. J.R. Leininger, D.V.M., Ph.D.
G.S. Travlos, D.V.M. National Toxicology Program
M:K. Valiant, B.S., M.T. J. Mahler, D.V.M.
K.L. Witt, M.S., ILS, Inc. National Toxicology Program
A. Nyska, D.V.M.
Battelle Columbus Laboratories National Toxicology Program
A. Radovsky, D.V.M., Ph.D.
Conducted studies, evaluated pathology findings
National Toxicology Program
D. Wolf, D.V.M., Ph.D.
M.R. Hejtmancik, Ph.D., Principal Investigator
Chemical Industry Institute ofToxicology
M.J. Ryan, D.V.M., Ph.D.
A.W. Singer, D.V.M. Evaluated slides and prepared pathology reports ofallyl alcohol
J.D. Toft, II, D.V.M., M.S. and acrolein (March 4, 1997)
Experimental Pathology Laboratories, Inc. J.C. Seely, D.V.M., Chairperson
PATHCO, Inc.
Provided pathology review
S. Botts, M.S., D.V.M., Ph.D.
Experimental Pathology Laboratories, Inc.
J.F. Hardisty, D.V.M., Principal Investigator J.R. Leininger, D.V.M., Ph.D.
S. Botts, M.S., D.V.M., Ph.D. National Toxicology Program
J. Mahler, D.V.M.
Constella Group, Inc. National Toxicology Program
A. Radovsky, D.V.M., Ph.D.
Provided statistical analyses
National Toxicology Program
P.W. Crockett, Ph.D., Principal Investigator D. Wolf, D.V.M., Ph.D.
L.J. Betz, M.S. Chemical Industry Institute ofToxicology
K.P. McGowan, M.B.A.
Biotechnical Services, Inc.
J.T. Scott, M.S.
Prepared Toxicity Study Report
S.R. Gunnels, M.A., Principal Investigator
J.M. Gregory, B.S.
D.C. Serbus, Ph.D.
W.D. Sharp, B.A., B.S.
P.A. Yount, B.S.
3
PEER REVIEW
The draft report on the toxicity studies of allyl acetate, allyl alcohol, and acrolein was evaluated by the reviewers
listed below. These reviewers serve as independent scientists, not as representatives of any institution, company, or
governmental agency. In this capacity, reviewers determine if the design and conditions of these NTP studies are
appropriate and ensure that the toxicity study report presents the experimental results and conclusions fully and
clearly.
James P. Kehrer, Ph.D. Mary Vore, Ph.D.
College ofPharmacy Graduate Center for Toxicology
University ofTexas University of Kentucky
Austin, TX Lexington, KY
4
CONTENTS
ABSTRACT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

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

Procurement and Characterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Preparation and Analysis of Dose Formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Toxicity Study Designs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Statistical Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Quality Assurance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Genetic Toxicology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

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

Rats . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Mice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55

Genetic Toxicology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

DISCUSSION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69

REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73

APPENDIXES
Appendix A Summary ofNonneoplastic Lesions in Rats . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-I

Appendix B Summary ofNonneoplastic Lesions in Mice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-1

Appendix C Clinical Pathology Results.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . C-1

Appendix D Organ Weights and Organ-Weight-to-Body-Weight Ratios . . . . . . . . . . . . . . . . . . . . . . . . . . D-1

Appendix E Reproductive Tissue Evaluations and Estrous Cycle Characterization . . . . . . . . . . . . . . . . . . E-1

Appendix F 3-Hydroxypropyl Mercapturic Acid Concentrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . F-1

Appendix G Genetic Toxicology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . G-1

Appendix H Chemical Characterization and Dose Formulation Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . H-1
5
ABSTRACT

Allyl Acetate Allyl Alcohol Acrolein
O
||
CH -C-O-CH -CH=CH CH =CH-CH -OH CH =CHCHO
3 2 2 2 2 2
CAS Number: 591-87-7 107-18-6 107-02-8
Molecular Weight: 100.12 58.08 56.06
Synonyms and Trade Names:
allyl alcohol, acetate; allylic alcohol; acraldehyde;
3-acetoxy-1-propene; 3-hydroxypropene; allyl aldehyde;
3-acetoxy-propene 1-propenol-3; 2-propenol; 2-propenal
2-propenyl alcohol;
Shell unkrautted A;
Weed Drench;
vinyl carbinol
Allyl acetate, allyl alcohol, and acrolein are used in the manufacture of detergents, plastics, pharmaceuticals, and
chemicals and as agricultural agents and food additives. Male and female F344/N rats and B6C3F mice received allyl
1
acetate, allyl alcohol, or acrolein by gavage for 14 weeks. Genetic toxicology studies were conducted in Salmonella
typhimurium, Drosophila melanogaster, cultured Chinese hamster ovary cells, rat bone marrow erythrocytes, and
mouse peripheral blood erythrocytes.
Groups of 10 male and 10 female rats were administered 0, 6, 12, 25, 50, or 100 mg allyl acetate/kg body weight, 0, 1.5,
3, 6, 12, or 25 mg/kg allyl alcohol, or 0, 0.75, 1.25, 2.5, 5, or 10 mg/kg acrolein in 0.5% methylcellulose by gavage,
5 days per week for 14 weeks. Groups of 10 male and 10 female mice were administered 0, 8, 16, 32, 62.5, or
125 mg/kg allyl acetate, 0, 3, 6, 12, 25, or 50 mg/kg allyl alcohol, or 0, 1.25, 2.5, 5, 10, or 20 mg/kg acrolein in
0.5% methylcellulose by gavage, 5 days per week for 14 weeks.
In the allyl acetate rat study, all males and females in the 100 mg/kg groups died or were killed moribund by day 8; there
were no other deaths. In the allyl alcohol study, all rats survived to the end of the study except one 6 mg/kg female.
In the acrolein rat study, eight males and eight females in the 10 mg/kg groups died by week 9 of the study. Two males
in the 2.5 and 5 mg/kg groups and one or two females in the 1.25, 2.5, and 5 mg/kg groups also died early; two of these
deaths were gavage accidents. In the allyl acetate mouse study, all males and females in the 125 mg/kg group died
during the first week of the study. All other early deaths, except five 62.5 mg/kg males and one 32 mg/kg female, were
gavage accidents. In the allyl alcohol mouse study, one 50 mg/kg female died due to a gavage accident; all other
6 Allyl Acetate, Allyl Alcohol, and Acrolein, NTP TOX 48
animals survived to the end ofthe study. In the acrolein mouse study, all males and females administered 20 mg/kg died
during the first week ofthe study. All other early deaths, except one male and one female administered 10 mg/kg, were
umelated to chemical administration.
The concentration of3-hydroxypropyl mercapturic acid (3-HPM) in the urine ofrats and mice was determined after the
first dose of chemical and at the end ofthe 14-week study. At both time points, the concentrations of3-HPM in the
urine ofanimals that received allyl acetate or allyl alcohol increased linearly with dose. In animals dosed with acrolein,
the concentrations of3-HPM exhibited a nonlinear increase with dose at the first time point. At the end ofthe study,
the concentration of 3-HPM in the urine of animals dosed with acrolein was linear with dose except at the highest
concentration administered. Since urine volumes were not recordedduring the urine collection, complete quantitation
ofthese data was not possible.
The final mean body weights and mean body weight gains ofmale rats administered 12 or 50 mg/kg allyl acetate and
ofmale and female rats administered 10 mg/kg acrolein were significantly less than those ofthe vehicle controls. The
mean body weight gain of male mice in the 50 mg/kg group in the allyl alcohol study was also less than that of the
vehicle controls. Final mean bodyweights and mean body weight gains ofdosed female rats and male and female mice
in the allyl acetate studies, male and female rats and female mice in the allyl alcohol studies, and male and female mice
in the acrolein studies were generally similar to those ofthe respective vehicle controls.
Clinical findings related to allyl acetate administration included pallor, eye or nasal discharge, ruffled fur, lethargy,
diarrhea, and thinness among rats in the 100 mg/kg groups and lethargy, abnormal breathing, thinness, and ruffled fur
among mice that died early. In the acrolein study, clinical findings included abnormal breathing, eye ornasal discharge,
ruffled fur, thinness, and lethargy in rats in the 10 mg/kg groups.
The liver weights ofmale rats administered 25 mg/kg allyl alcohol, female rats administered 50 mg/kg allyl acetate or
5 or I 0 mg/kg acrolein, and male mice administered 10 mg/kg acrolein were significantly greater than those of the
vehicle controls. Female rats administered 10 mg/kg acrolein had significantly lower absolute and relative thymus
weights than did the vehicle controls.
Female rats administered 25 mg/kg allyl alcohol spent more time in diestrus and less time in metestrus than the vehicle
controls. The estrous cycles offemale mice dosed with 16 or 32 mg/kg allyl acetate were significantly longer than that
of the vehicle controls.
Allyl Acetate, Allyl Alcohol, and Acrolein, NTP TOX 48 7
Gross lesions related to allyl acetate treatment were observed in the liver, forestomach, and thorax/abdomen of male
and female rats in the I 00 mg/kg groups. Microscopically, the incidences of forestomach squamous epithelial
hyperplasia were significantly increased in male rats administered I2 mg/kg or greater, female rats administered 25 or
50 mg/kg, male mice administered 32 or 62.5 mg/kg, and female mice administered 16, 32, or 62.5 mg/kg. Forestomach
necrosis, hemorrhage, and inflammation were present in most rats in the I 00 mg/kg groups, and the incidence of
hemorrhage in I25 mg/kg male mice was increased; male mice in the 62.5 and I25 mg/kg groups and I25 mg/kg female
mice had significantly increased incidences of glandular stomach hemorrhage. Increased incidences of several liver
lesions occurred in male or female rats administered 50 or I 00 mg/kg, and to a lesser extent in 25 mg/kg rats,
62.5 mglkg male mice, and I25 mg/kg male and female mice.
Bone marrow hyperplasia, hemorrhage or depletion in the mediastinal, mandibular, and mesenteric lymph nodes,
hemorrhage and necrosis of the thymus, and hematopoietic cell proliferation of the red pulp were also observed in
100 mglkg rats. Increased incidences ofnecrosis in the mandibular and mesenteric lymph nodes, spleen, and thymus
were observed in 62.5 and I25 mg/kg mice.
Male and female rats administered 6 mg/kg allyl alcohol or greater and male and female mice administered 12 mg/kg
allyl alcohol or greater had significantly increased incidences ofsquamous hyperplasia ofthe forestomach epithelium.
Female rats in the 25 mg/kg group had significantly increased incidences of bile duct hyperplasia and periportal
hepatocyte hypertrophy in the liver. Incidences of portal cytoplasmic vacuolization were significantly increased in
50 mg/kg male mice and female mice in the 25 and 50 mg/kg groups.
Gross lesions related to acrolein treatment were observed in the forestomach and glandular stomach ofmale and female
rats in the I 0 mg/kg groups and 20 mg/kg female mice. Microscopically, the incidences of squamous hyperplasia of
the forestomach epithelium were significantly increased in male rats in the 5 and 10 mg/kg groups, female rats
administered 2.5 mg/kg or greater, and male and female mice administered 2.5, 5, or 10 mg/kg. Male and female rats
in the 10 mg/kg groups and 20 mg/kg male and female mice had significantly increased incidences ofglandular stomach
hemorrhage. Female mice in the 20 mglkg group also had significantly increased incidences of glandular stomach
inflammation and epithelial necrosis.
Allyl acetate was mutagenic inS. typhimurium strains TAIOO and TAI535, in the absence ofS9 activation. With S9,
no mutagenicity was detected in these two strains; negative results were obtained in strains TA97 and TA98, with and
without S9. Allyl alcohol was not mutagenic in four strains ofS. typhimurium, with or without S9 metabolic activation.
Acrolein, tested in a preincubation protocol, was weakly mutagenic inS. typhimurium strain TAl 00 in the presence of
10% induced rat liver S9. Equivocal results were obtained in strains TAIOO and TAI535 with IO% induced hamster
liver S9. Negative results were obtained with TA97, TA98, and TA1538 under all test conditions, and acrolein gave
negative results in all fourS. typhimurium strains tested for mutation induction under a vapor protocol. No induction
8 Allyl Acetate, Allyl Alcohol, and Acrolein, NTP TOX 48
ofmicronuclei was noted in bone marrow erythrocytes ofmale rats administered allyl acetate by gavage three times at
24-hour intervals. No significant increases in micronucleated erythrocytes were noted in bone marrow samples from
male rats administered allyl alcohol by intraperitoneal injection for 3 days. A small, but significant increase in the
frequency of micronucleated normochromatic erythrocytes was observed in the peripheral blood of female mice
administered allyl acetate by gavage for 14 weeks; no increase was observed in male mice. No increases in the
frequencies of micronucleated normochromatic erythrocytes were observed in the peripheral blood ofmale or female
mice administered allyl alcohol or acrolein by gavage for 14 weeks. Acrolein induced sister chromatid exchanges in
cultured Chinese hamster ovary cells in the absence, but not the presence, of S9; it did not induce chromosomal
aberrations, with orwithout S9. Results ofthree independent Drosophila melanogaster sex-linked recessive lethal tests
in which acrolein was administered to adult flies via feeding or injection and to larvae via feeding were negative.