ASTM_E_1073_-_01.pdf

Designation:E 1073 01Standard Test Method forObtaining a Pharmacological Profile with Mice1This standard is issued under the fixed designation E 1073;the number immediately following the designation indicates the year oforiginal adoption or,in the case of revision,the year of last revision.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 is designed as a simple and inexpen-sive initial screening procedure for new compounds withunknown pharmacological properties,or for the comparativebioassay of new members of a chemical series with classreference standards.The test method,which is applicable tomost pharmacologically active compounds including pesti-cides,will properly rank order both acute lethality and potencywith a minimum expenditure of test material.It is intended asthe first step in a multi-tiered development program.1.2 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.Summary of Test Method2.1 Mice are injected intravenously with a dose of the testmaterial and then observed for reaction signs,first those thatcan be detected by nonmanipulative tests,and then thosesensed during a series of manipulative tests.2.2 Two technicians are required to perform the experiment.One injects the mice and records the data,while the otherconducts the experiment at regularly scheduled time intervalsand dictates the findings.2.3 The results obtained using this procedure indicate theapproximate median lethal dose(LD50),the approximatemedian level of nonlethal reactions(MED50),the reactionsigns elicited at each dose level tested,and the degree ofseverity of those signs.In contrast to the more commonly usedmedian effective dose(ED50),the MED50 value may havebeen generated by one or more responses at the same dose,andthus does not imply the 50%level for a specific reaction sign.2.4 By judicious selection of acceptance criteria,the elimi-nation rate for compounds tested using this procedure can betailored to any desired level.3.Significance and Use3.1 This test method is designed as an initial screeningprocedure for the selection of compounds worthy of moredetailed study.3.2 This test method is applicable to the study of most drugsand chemicals,and will properly estimate both lethal andminimally effective dose levels.Although it is designed for thestudy of single components,it can be used to study thecomparative toxicity of mixtures or formulations.The methodmay not be applicable to oily substances which cause embo-lism upon injection.3.3 This test method requires only small quantities of testmaterials(approximately 1 g),a fact that enhances its utility asthe initial biological study for newly synthesized substances.3.4 It is equally economical in its requirements for equip-ment,space,personnel,and animals.Only a small laboratory,simple test equipment,and two technicians are needed toconduct the experiments.Furthermore,an average of onlythirty mice are required to conduct the entire test method.3.5 The procedure is applicable to a wide variety of mate-rials.When results of this test were compared with those frommore detailed and specific animal tests,a high degree ofcorrelation was obtained.Further evidence of the utility of thetest was demonstrated by the fact that a high correlation of rankorder potencies was found for a series of anticholinergicsstudied in both mice and men.4.Apparatus4.1 All of the apparatus used for this test method is simpleand inexpensive.It can be made in any well-equipped labora-tory workshop or,in some cases,obtained commercially.4.2 The major components of the apparatus are illustrated inFigs.1-5.4.3 Syringes and Needles:4.3.1 One-quarter millilitre glass Tuberculin syringes,fittedwith 0.75 in.(19 mm),27-gage needles are recommended forinjection of all solutions except undiluted polyethylene glycol(PEG)solutions.In the latter case,24-gage needles must beused to compensate for the viscosity of the solvent,and 50-Lsyringes2,3are required to measure accurately the small vol-umes that are administered.1This test method is under the jurisdiction of ASTM Committee E-35 onPesticides and is the direct responsibility of Subcommittee E35.26 on Safety to Man.Current edition approved Oct.10,2001.Published November 2001.Originallypublished as E 1073 85.Last previous edition E 1073 91(1996)1Copyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959,United States.4.3.2 Although disposable syringes could be used for mostof this work,calibrations on the glass syringes are moreaccurate.Furthermore,the possibility of a reaction between theplastic or rubber parts of disposable syringes with the testsolutions,especially those c