ASTM E1163-10(2019)
(Test Method)Standard Test Method for Estimating Acute Oral Toxicity in Rats
Standard Test Method for Estimating Acute Oral Toxicity in Rats
SIGNIFICANCE AND USE
5.1 This test method is of principal value in minimizing the number of animals required to estimate the acute oral toxicity (LD50). It also incorporates measures of variance (95 % CI) and a slope from which to make relative toxicity comparisons.
5.2 This test method is inappropriate for materials typically producing death two or more days after administration of the test compound unless the observation time between dosages is increased. This test method can be successfully applied, however, for materials producing only an occasional death two or more days after administration.
5.3 The LD50 is valuable as a measure of the relative acute toxicity of a material and can be used to make an estimate of potential hazard to humans when pesticides, other chemicals, or mixtures are ingested.
5.4 This test method allows for observation of signs of toxicity in addition to mortality. This information can be useful in planning additional toxicity testing.
SCOPE
1.1 This test method determines the lethality (LD50 value, slope and 95 % confidence interval (CI)) and signs of acute toxicity from a material using a limited number of rats. The technique used in this test method is referred to as the “Stagewise, Adaptive Dose Method.”2 This test method is an alternative to the classical LD50 test and is applicable to both liquids and solids.
1.2 This test method is not recommended for test materials which typically produce deaths beyond two days postdosing.
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
General Information
Relations
Standards Content (Sample)
This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E1163 − 10 (Reapproved 2019)
Standard Test Method for
Estimating Acute Oral Toxicity in Rats
This standard is issued under the fixed designation E1163; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3.1.1 delayed death—an animal which does not die or
appear moribund within 24 h but dies later during the obser-
1.1 This test method determines the lethality (LD50 value,
vation period.
slope and 95% confidence interval (CI)) and signs of acute
3.1.2 gavage—forcedoraldosing,asbyatubethatispassed
toxicity from a material using a limited number of rats. The
down the throat to the stomach.
technique used in this test method is referred to as the
3.1.3 LD50—thestatisticallyderivedestimateofthedoseof
“Stagewise, Adaptive Dose Method.” This test method is an
atestsubstancethatwouldbeexpectedtocause50%mortality
alternative to the classical LD50 test and is applicable to both
to the test population under the specified test conditions.
liquids and solids.
3.1.4 moribund—at the point of death or extinction.
1.2 This test method is not recommended for test materials
3.1.5 pharmacotoxic—grossphysiologicalsignsinresponse
which typically produce deaths beyond two days postdosing.
to a toxic material.
1.3 This standard does not purport to address all of the
3.1.6 signs of toxicity—objective, observable evidence of
safety concerns, if any, associated with its use. It is the
toxicity.
responsibility of the user of this standard to establish appro-
3.1.7 suspension—a mixture in which very small particles
priate safety, health, and environmental practices and deter-
remain suspended without dissolving.
mine the applicability of regulatory limitations prior to use.
3.1.8 toxicity—poisonous quality.
1.4 This international standard was developed in accor-
dance with internationally recognized principles on standard-
4. Summary of Test Method
ization established in the Decision on Principles for the
Development of International Standards, Guides and Recom- 4.1 Three to five different doses of the target compound are
mendations issued by the World Trade Organization Technical
selected such that the doses span the entire dose response
Barriers to Trade (TBT) Committee. curve, with separation between the doses to be equal log
intervals. One to two animals are given each dose as the first
2. Referenced Documents
stage of the study.After 24 to 48 h, the responses to each dose
are observed and used in determining the doses and animal
2.1 ASTM Standards:
numbers in the next stage of dosing.
E609Terminology Relating to Pesticides
IEEE/ASTM SI10Standard for Use of the International
4.2 The second and subsequent stages have one to four
System of Units (SI) (the Modernized Metric System)
doses with one to three animals at each dose. Doses for
subsequent stages are selected based on the estimates of the
3. Terminology
dose response distribution parameters and the uncertainties of
3.1 Definitions:
theseestimates.Thedoseresponsecurveanditsparametersare
updated after each stage and dosing will stop when the 95 %
confidence interval for the LD50 satisfies the following stop-
This test method is under the jurisdiction of ASTM Committee E50 on
ping rule: (upper 95 % CI – lower 95 % CI)/ (2 × LD50) < =
Environmental Assessment, Risk Management and Corrective Action and is the
0.40).
direct responsibility of Subcommittee E50.47 on Biological Effects and Environ-
mental Fate.
4.3 The slope, LD50 and its 95 % confidence interval are
Current edition approved Feb. 1, 2019. Published February 2019. Originally
calculated using the methods Feder. No more than the use of
approved in 1987. Last previous edition approved in 2016 as E1163–10(2016).
DOI: 10.1520/E1163-10R19. 30 animals is recommended and shall constitute an additional
Feder,P.I.,StatisticalDesignConsiderationsforStagewise,stagewise,adaptive
stopping rule.
doseallocationindoseresponsivestudies.In:Peace,KarlE.,ed.Biopharmaceutical
sequential statistical applications. New York: Marcel Dekker 1992.
5. Significance and Use
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
5.1 This test method is of principal value in minimizing the
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. number of animals required to estimate the acute oral toxicity
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E1163 − 10 (2019)
(LD50). It also incorporates measures of variance (95 % CI) liquid form, that is liquid, solution, suspension, or emulsion,
and a slope from which to make relative toxicity comparisons. suitable for administration by gavage.
5.2 This test method is inappropriate for materials typically
9.2 Thetestmaterialshallbeatthesametemperatureasthat
producing death two or more days after administration of the
of the room in which the test is conducted at the time of
test compound unless the observation time between dosages is
administration to the animals.
increased. This test method can be successfully applied,
however,formaterialsproducingonlyanoccasionaldeathtwo
10. Procedure
or more days after administration.
10.1 Deprive the animals of food for 12 to 18 h before
5.3 The LD50 is valuable as a measure of the relative acute
administering the test substance.
toxicity of a material and can be used to make an estimate of
potential hazard to humans when pesticides, other chemicals, 10.2 Weight of each rat and calculate the dose according to
thisbodyweighttogivethespecifiedquantityoftestsubstance
or mixtures are ingested.
per unit of body weight.
5.4 This test method allows for observation of signs of
toxicityinadditiontomortality.Thisinformationcanbeuseful
10.3 Record all information necessary to document animal
in planning additional toxicity testing.
weights and volume of test substance administered to each
animal.
6. Apparatus
10.4 Dose four to six animals in the first stage each at a
6.1 Syringe, and an oral dosing needle or rubberized cath-
differentdosespanningtheestimateddoseresponsecurve(0%
eter to gavage the test compound is required.
lethality to 100 % lethality). Gavage animals using an oral
dosing needle or rubberized tubing. Observe each animal for a
7. Test Animals
minimum of 24 h. Use the methods of Feder et al (1992) to
7.1 Albino female rats weighing 190 to 300 g prefasted are
estimate the dose response curve and its parameters. Check to
used. A non in-bred rat such as the Sprague-Dawley strain is
see if the stopping rule (defined in 4.2) is met. If not, then
generally preferred. Female rats are preferred because histori-
proceed to the second stage of dosing. Return the animals to
cal data indicate that females in most instances have lower
either ad libitum or 2 to 3-h feeding immediately after dosing.
LD50 values than males.
Return the animal to either ad libitum or 2 to 3-h feeding
7.2 An additional test may be conducted with male rats, b
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