ASTM E1163-1998(2002) Standard Test Method for Estimating Acute Oral Toxicity in Rats《评定剧毒口服灭鼠剂的试验方法》.pdf

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1、Designation: E 1163 98 (Reapproved 2002)Standard Test Method forEstimating Acute Oral Toxicity in Rats1This standard is issued under the fixed designation E 1163; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last rev

2、ision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) indicates an editorial change since the last revision or reapproval.1. Scope1.1 This test method determines the lethality (LD50 value)and signs of acute toxicity from a material using a limitednumber of ra

3、ts. The technique used in this test method isreferred to as the “Up-and-Down Procedure.”2This testmethod is an alternative to the classical LD50 test and isapplicable to both liquids and solids.1.2 This test method is not recommended for test materialswhich typically produce deaths beyond 2 days pos

4、tdosing.1.3 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use.2. Refe

5、renced Documents2.1 ASTM Standards:3E 609 Terminology Relating to PesticidesIEEE/ASTM SI 10 Standard for Use of the InternationalSystem of Units (SI) (the Modernized Metric System)3. Terminology3.1 Definitions:3.1.1 delayed deathan animal which does not die orappear moribund within 24 h but dies lat

6、er during the obser-vation period.3.1.2 gavageforced feeding, as by a tube that is passeddown the throat to the stomach.3.1.3 LD50the statistically derived estimate of the dose ofa test substance that would be expected to cause 50 % mortalityto the test population under the specified test conditions

7、.3.1.4 moribundat the point of death or extinction.3.1.5 suspensiona mixture in which very small particlesremain suspended without dissolving.3.1.6 toxicitypoisonous quality.3.1.7 signs of toxicityobjective, observable evidence oftoxicity.3.1.8 pharmacotoxicgross physiological signs in responseto a

8、toxic material.4. Summary of Test Method4.1 Female rats are dosed one at a time starting the firstanimal at the best estimate of the LD50. If this animal survives,then the next animal receives a higher dose; but if the firstanimal dies, the next animal receives a lower dose. If possible,subsequent d

9、oses are adjusted by a constant multiplicativefactor, for example 1.3. The dose for each animal is adjusted upor down depending upon the outcome for the previous animal.4.2 The dosing is repeated as above until four animals havebeen dosed after reversal of the initial outcome.4.3 The LD50 is calcula

10、ted using the maximum likelihoodmethod.45. Significance and Use5.1 This test method is of principal value in minimizing thenumber of animals required to estimate the acute oral toxicity(LD50).5.2 This test method is inappropriate for materials typicallyproducing death 2 or more days after administra

11、tion of the testcompound unless the observation time between dosages isincreased. This test method can be successfully applied,however, for materials producing only an occasional death 2 ormore days after administration.5.3 The LD50 is valuable as a measure of the relative acutetoxicity of a materia

12、l and can be used to make an estimate ofpotential hazard to humans when pesticides, other chemicals,or mixtures are ingested.5.4 This test method allows for observation of signs oftoxicity in addition to mortality. This information can be usefulin planning additional toxicity testing.6. Apparatus6.1

13、 Syringe,5and an oral dosing needle or rubberizedcatheter to gavage the test compound are required.1This test method is under the jurisdiction of ASTM Committee E35 onPesticides and Alternative Control Agents and is the direct responsibility ofSubcommittee E35.26 on Safety to Man.Current edition app

14、roved Oct. 10, 2002. Published March 2003. Originallyapproved in 1987. Last previous edition approved in 1998 as E 1163 98.2Bruce, R. D., “An Up-and-Down Procedure for Acute Toxicity Testing,”Fundamental and Applied Toxicology, Vol 5, 1985, pp. 151157.3For referenced ASTM standards, visit the ASTM w

15、ebsite, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.4Finney, D. J., Probit Analysis, 3rd ed., Cambridge University Press, Cambridge,England, 1971, pp. 5080.5Hami

16、lton Milliliter syringes available from Hamilton Co., Reno, NV have beenfound suitable for this purpose.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.7. Test Animals7.1 Albino female rats weighing 190 to 300 g prefasted areused. A

17、non in-bred rat such as the Sprague-Dawley strain isgenerally preferred. Female rats are preferred because histori-cal data indicate that females in most instances have lowerLD50 values than males.67.2 An additional test may be conducted with male rats, butit is not necessary, unless it is suspected

18、 that the substance ismore toxic to males than females.8. Pretest Conditioning8.1 Examine each test animal on arrival for overt signs ofdisease, and condition to the environment for a minimum of 7days. Select animals that have not been used for any other tests.8.2 Maintain animals during pretest and

19、 test periods inaccordance with accepted laboratory practices for the care andhandling of test animals.8.3 Identify each animal with an ear tag or other suitablemeans.8.4 During acclimation, observe the animals for adversehealth effects. Eliminate any animal(s) demonstrating signs ofspontaneous dise

20、ase prior to the start of the study. Use onlyanimals judged to be healthy.8.5 The animals are housed individually. Rat chow or theequivalent and water are to be available ad libitum after dosing.9. Sample Preparation9.1 Because of the great variety of physical characteristicsand formulations of chem

21、icals and pesticides, it is not possibleto stipulate how the test material should be prepared. The onlycriterion that can be specified is that the material must be inliquid form, that is liquid, solution, suspension, or emulsion,suitable for administration by gavage.9.2 The test material shall be at

22、 the same temperature as thatof the room in which the test is conducted at the time ofadministration to the animals.10. Procedure10.1 Weigh the animals and deprive of food for 18 to 20 hbefore administering the test substance.10.2 Determine fasted body weight of each rat and calculatethe dose accord

23、ing to this body weight to give the specifiedquantity of test substance per unit of body weight.10.3 Record all information necessary to document animalweights and volume of test substance administered to eachanimal.10.4 Gavage one animal using an oral dosing needle orrubberized tubing. Return the a

24、nimal to either ad libitum or 2to 3-h feeding immediately after dosing.10.5 Observe the animal for mortality and pharmacotoxicsigns periodically for the first 4 h after dosing (at least onceduring the first 30 min) and daily thereafter for a total of 7days.10.6 Pharmacotoxic signs most frequently se

25、en are as fol-lows: respiratory rate increase or decrease, hypoactivity, pros-tration, ataxia, unkept appearance, body tremors, blanching,gasping, diarrhea, lethargy, chromodacryorrhea, and red excre-tion around nares.10.7 Dose one animal at a time starting at the estimatedLD50. Observe each animal

26、for a minimum of 24 h. If theanimal dies or appears moribund with symptoms such asshallow, labored or irregular respiration, muscular weakness ortremors, absence of voluntary responses to external stimuli,cyanosis and coma, decrease the dose for the next animal. If theanimal survives and appears hea

27、lthy, increase the dose for thenext animal. If feasible, use a dose progression factor of 1.3.10.8 After reaching the reversal of the initial outcome (thatis the point where an increasing dose pattern is required to bedecreased by a death or a decreasing dose pattern is required tobe increased by a

28、survival), dose an additional four animals inthe up-down procedure and then stop.10.9 If ten animals have been dosed with increasing dosagesand no deaths have occurred within 24 h but delayed deaths areobserved in three or more animals, stop the procedure. In thiscase, report that an LD50 could not

29、be determined using thisprocedure.10.10 Record all pharmacotoxic symptoms and time ofdeath. Perform a gross necropsy on all animals that die.10.11 Weigh all surviving animals on day 7 and perform agross necropsy. The necropsy should entail a macroscopicinspection of all visceral organs. Record all f

30、indings.11. Test Options11.1 If deemed necessary, perform the test concurrently andindependently in both sexes using the same strain and bodyweight.11.2 To ascertain the lethality in males, dose six male rats atthe determined LD50 value of the females. Report the LD50 asfollows:11.2.1 If the number

31、of decreased males is none, report thatwith 95 % confidence the LD50 value for males exceeds theadministered dose.11.2.2 If one or two animals die, report that the estimatedLD50 value for males exceeds the administered dose.11.2.3 If three animals die, report that the estimated LD50value for males e

32、quals the administered dose.11.2.4 If four or five animals die, report that the estimatedLD50 value for males is less than the administered dose.11.2.5 If six of six animals die, execute the up-and-downprocedure described in this test method using male rats.12. Calculation of Results12.1 If all the

33、dead animals have higher doses than all thelive animals, then the LD50 is between the doses for the liveand dead animals. If needed, the binomial test can be used tocalculate the probability that the LD50 is between the lowestdose that killed all of the animals and the highest dose thatkilled none o

34、f the animals. If this probability is more than95 %, then use the geometric mean of the two doses as anapproximate LD50.12.1.1 Additional toxicity tests should not be conducted ifthe existing test provides answers to all of the practicalquestions that need to be answered concerning the LD50.6Dixon,

35、W. J., “The Up-and-Down Method for Small Samples,” Journal ofAmerican Statistics Association, Vol 60, 1965, pp. 967978.E 1163 98 (2002)212.2 If the live and dead animals have only one dose incommon and all other dead animals have higher doses and allother live animals have lower doses, then LD50 may

36、 approxi-mate their common dose. Additional testing should only beperformed when it is determined that there is a practical need.12.3 No estimate of the LD50 should be made when theslope of the maximum likelihood probit curve is zero. Thisevent will happen when (SNA)(SARA)=(SRA)(SANA),where A denote

37、s the dose, NAdenotes the number of animalsreceiving dose A, RAdenotes the number of animals receivingdose A that die and the summations are over all administereddoses.12.4 If none of the above situations occur, then calculate theLD50 using the maximum likelihood method.4This calculationmay also be

38、performed using the SAS7or BMDP8computerprogram packages or other suitable computer programs. Othermethods, including nonparametric methods, can be used,especially if probit and logit methods do not fit the data.13. Report13.1 The report shall contain such information as testspecies and source and s

39、ex of animals.13.2 The report should include study initiation and termi-nation dates, individual body weights, dose levels, dose ad-ministered, return to feeding time, mortality, pharmacotoxicsigns, gross necropsy results, and LD50, if determined.14. Precision and Bias14.1 A precision and bias state

40、ment cannot be made at thistime.15. Keywords15.1 chemicals; LD50; oral toxicity; pesticides; ratsASTM International takes no position respecting the validity of any patent rights asserted in connection with any item mentionedin this standard. Users of this standard are expressly advised that determi

41、nation of the validity of any such patent rights, and the riskof infringement of such rights, are entirely their own responsibility.This standard is subject to revision at any time by the responsible technical committee and must be reviewed every five years andif not revised, either reapproved or wi

42、thdrawn. Your comments are invited either for revision of this standard or for additional standardsand should be addressed to ASTM International Headquarters. Your comments will receive careful consideration at a meeting of theresponsible technical committee, which you may attend. If you feel that y

43、our comments have not received a fair hearing you shouldmake your views known to the ASTM Committee on Standards, at the address shown below.This standard is copyrighted by ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959,United States. Individual reprints (si

44、ngle or multiple copies) of this standard may be obtained by contacting ASTM at the aboveaddress or at 610-832-9585 (phone), 610-832-9555 (fax), or serviceastm.org (e-mail); or through the ASTM website(www.astm.org).7SAS(r) Proprietary Software is available from SAS Institute Inc., Cary, NC.8Dixon, W. J., ed., BMDP Statistics Software, University of California Press,Berkely, CA.E 1163 98 (2002)3