ASTM E1103-96

Standard Test Method for Determining Subchronic Dermal Toxicity

SCOPE
1.1 This test method describes a procedure for the assessment and evaluation of the toxic characteristics of a test substance that is applied daily to the skin of experimental animals for 90 days.  
1.2 This test method is not capable of determining effects that have a long latency period (for example, carcinogenicity) or are life shortening.  
1.3 This test method is intended primarily to be used with rats, guinea pigs, or rabbits. Other species may be used with appropriate modifications.
1.4 This standard does not purport to address the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

General Information

• Status: Historical
• Publication Date: 31-Dec-1999
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents

Relations

• Referred By: ASTM E1302-23 - Standard Guide for Acute Animal Toxicity Testing of Water-Miscible Metalworking Fluids
• Effective Date: 01-Jan-2000

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
Designation: E 1103 – 96
Standard Test Method for
Determining Subchronic Dermal Toxicity
This standard is issued under the fixed designation E 1103; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
1. Scope stance Control Act, Part 792, Good Laboratory Practice
Standards
1.1 This test method describes a procedure for the assess-
ment and evaluation of the toxic characteristics of a test
3. Terminology
substance that is applied daily to the skin of experimental
3.1 Definitions—For definitions of terms used in this test
animals for 90 days.
method, see Definitions E 609.
1.2 This test method is not capable of determining effects
3.2 Definitions of Terms Specific to This Standard:
that have a long latency period (for example, carcinogenicity)
3.2.1 dose, dosage, n—the quantity of a substance applied
or are life shortening.
per unit treated or applied to or entered into organism. This is
1.3 This test method is intended primarily to be used with
expressed as the weight of the test substance per unit weight of
rats, guinea pigs, or rabbits. Other species may be used with
test animal (mg/kg).
appropriate modifications.
3.2.2 no observed adverse effect dose (NOAED), n—the
1.4 This standard does not purport to address the safety
highest tested dose of a substance at which the measured
concerns, if any, associated with its use. It is the responsibility
biological variables of a specific group under test conditions
of the user of this standard to establish appropriate safety and
show no statistically significant dose-related adverse difference
health practices and determine the applicability of regulatory
from the control treatment group.
limitations prior to use.
3.2.3 nulliparous, adj—having never borne an offspring.
2. Referenced Documents 3.2.4 test substance, n—pesticide or other material (ele-
ment, chemical compound, formulation, known mixture) ad-
2.1 ASTM Standards:
2 ministered orally for the purpose of determining subchronic
E 609 Definitions of Terms Relating to Pesticides
oral toxicity.
E 758 Test Method for Mammalian Acute Percutaneous
Toxicity
4. Summary of Test Method
2.2 Federal Standards:
4.1 The test substance is applied daily to the skin in
Title 21, Code of Federal Regulations (CFR), Food and
geometrically graduated doses to several groups of animals,
Drug Administration, Part 58, Good Laboratory Practice
one dose per group, for a period of 90 days. Generally at least
for Nonclinical Laboratory Studies
three dose levels with a control and, where appropriate, a
Title 40, Code of Federal Regulations (CFR), Environmen-
vehicle control group are employed.
tal Protection Agency, Part 798, Health Effects Testing
4.2 Animals that die during the test are necropsied. At the
Guidelines, Subpart C, Subchronic Exposure, Oral Toxic-
conclusion of the test, the surviving animals are sacrificed and
ity
necropsied and appropriate histopathological examinations
Title 40, Code of Federal Regulations (CFR), Environmen-
performed.
tal Protection Agency, Part 798, Health Effects Testing
4.3 A limit test with one dose of at least 1000 mg/kg body
Guidelines, Subpart B, General Toxicity Testing, Acute
3 weight can be conducted using the procedures described for
Oral Toxicity
this method if no toxicity would be expected based upon data
Title 40, Code of Federal Regulations (CFR), Environmen-
of structurally related compounds. If this test produces no
tal Protection Agency, Subchapter E, Pesticide Programs;
3 observable toxic effects, then a full study using three doses is
Part 160, Good Laboratory Practice Standards
not necessary.
Title 40, Code of Federal Regulations (CFR), Toxic Sub-
5. Significance and Use
This test method is under the jurisdiction of ASTM Committee E-35 on 5.1 This test method provides information on health hazards
Pesticides and is the direct responsibility of Subcommittee E35.26 on Safety to Man.
likely to arise from repeated exposure to a pesticide or other
Current edition approved Oct. 10, 1996. Published December 1996. Originally
chemical by the dermal route over a short time period. It may
published as E 1103 – 86. Last previous edition E 1103 – 91.
provide information needed to establish safety criteria for
Annual Book of ASTM Standards, Vol 11.05.
Available from U.S. Government Printing Office, Superintendent of Docu-
human exposure.
ments, Washington, DC 20402.
Copyright © ASTM, 100 Barr Harbor Drive, West Conshohocken, PA 19428-2959, United States.
E 1103
5.2 Signs of toxicity other than lethality can be observed. 9.2 Maintain the animals during pretest and test periods
5.3 This method can provide information on target organs according to accepted laboratory practices for the care and
and possible cumulative effects. It may aid in the selection of handling of animals.
doses for chronic studies. 9.3 Identify each animal with an ear tag or other suitable
means.
6. Hazards
9.4 During acclimation, observe the animals for respiratory
6.1 Minimize contact with all test substances, solutions, and
distress, diarrhea, emaciation, ocular and nasal discharges, skin
mixed diets with appropriate protective clothing, gloves, eye
lesions, and eye defects. Eliminate any animal demonstrating
protection, and so forth. The use of fume hoods and increased
signs of spontaneous disease prior to the start of the study. Use
ventilation in test rooms is necessary when handling volatile
only animals judged to be healthy. Animals on test should be
substances. Know information on acute mammalian toxicity
segregated in different rooms. Chow or the equivalent and
and special handling procedures before this method is used.
water are to be available ad libitum.
6.2 Ensure health and environmental safety prior to disposal
10. Dose Level and Dose Selection
of excess test substances, solutions, mixed diets, excreta, and
treated animals, and in accordance with all federal, state, and 10.1 At least three doses plus a control must be used. Doses
local regulations.
shall be spaced geometrically to produce test groups with a
6.3 Clean and rinse glassware, feeders, and other equipment range of toxic effects and mortality rates. The data should be
with volatile solvents only in well ventilated areas. The use of
sufficient to produce a dose-response effect.
fume hoods may be necessary when handling volatile sub- 10.1.1 If no information is available for establishing doses,
stances.
then a 14-day range-finding test should be conducted.
6.4 Consider periodic medical examinations for all person- 10.2 The highest dose should result in toxic effects but not
nel caring for animals or handling test substances.
produce severe skin irritation or an excessively high incidence
of fatalities that would prevent a meaningful evaluation.
7. Facilities Required
10.3 The lowest dose should not produce any evidence of
7.1 Test animals shall be individually housed in cages
toxicity. Where there is a reasonable estimate of human
meeting the requirements specified in the Guide for The Care
exposure, however, the lowest dose should exceed this value.
and Use of Laboratory Animals.
10.4 If more than one intermediate dose is used, then the
7.2 Animal holding rooms should be maintained at 70 6
doses should be spaced to provide a gradation of toxic effects.
3°F (21.1 6 2°C), with 50 6 5 % relative humidity for guinea
11. Procedure
pigs or rats and 43 6 3 % for rabbits, and a 12-h light-dark
cycle.
11.1 Preparation of Animal Skin:
7.3 Testing areas shall be maintained at the same tempera- 11.1.1 Clip the fur from the dorsal area of the trunk of the
ture and humidity as animal holding rooms.
test animals approximately 24 h before the test. Repeat
clipping or shaving as needed, usually at approximately weekly
8. Test Animals
intervals. When clipping or shaving the fur, take care to avoid
8.1 Rats, rabbits, or guinea pigs should be used. However
abrading the skin which could alter the permeability of the
albino rabbits are preferred because of their size, skin perme-
skin.
ability, and extensive data base.
11.1.2 Clear at least 10 % of the body surface for the
8.1.1 If another mammalian species is used, the investigator
application of the test substance.
should record the justification for the selection.
11.2 Application of the Test Substance:
8.2 Young adult animals should be used. The following
11.2.1 When testing liquids, apply the test substance as is or,
weight ranges at the start of the test are recommended.
if appropriate, diluted in a suitable solvent. If a solvent is
8.2.1 Rats—200 to 300 g.
employed, then include a solvent control group of treated
8.2.2 Rabbits—2 to 3 kg.
animals. Consider the influence of the solvent on penetration of
8.2.3 Guinea Pigs—350 to 450 g.
skin by the test substance.
8.3 Equal numbers of animals of each sex with healthy skin
11.2.2 When testing solids, pulverize the test substance, if
should be used at each dose. A minimum of ten animals per sex
appropriate, and moisten with water or a suitable vehicle to
per group is recommended.
ensure good contact with the skin. When a vehicle is used,
8.3.1 The females should be nulliparious and nonpregnant.
consider the influence of the vehicle on penetration of skin by
8.3.2 If interim or post sacrifices are planned additional
the test substance; include a vehicle control group of treated
animals should be included in the study.
animals.
11.2.3 Apply the test substance uniformly over an area that
9. Pretest Conditioning
is approximately 10 % of the total body surface area.
9.1 Examine the animals on arrival for overt signs of disease
11.2.4 During the exposure period, hold the test substance in
and condition them to the environment for a minimum of 14
contact with the skin with a porous gauze and nonirritating
days. Select animals that have not been used on any other tests.
tape. Place Elizabethan collars around th
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