1、Designation:E269411An American National StandardStandard Test Method forMeasurement of Adenosine Triphosphate in Water-MiscibleMetalworking Fluids1This standard is issued under the fixed designation E2694;the number immediately following the designation indicates the year oforiginal adoption or,in t
2、he case of revision,the year of last revision.A number in parentheses indicates the year of last reapproval.Asuperscript epsilon()indicates an editorial change since the last revision or reapproval.1.Scope*1.1 The method provides a protocol for capturing,extract-ing and quantifying the adenosine tri
3、phosphate(ATP)contentassociated with microorganisms found in water-miscible met-alworking fluids(MWF).1.2 The ATP is measured using a bioluminescence enzymeassay,whereby light is generated in amounts proportional tothe concentration of ATP in the samples.The light is producedand measured quantitativ
4、ely as relative light units(RLU)which are converted by comparison with an ATP standard andcomputation to pg ATP/mL.1.3 This method is equally suitable for use in the laboratoryor field.1.4 The method detects ATP concentrations in the range of4.0 pg ATP/mL to 400,000 pg ATP/mL.1.5 Providing interfere
5、nces can be overcome,biolumines-cence is a reliable and proven method for qualifying andquantifying ATP.The method does not differentiate betweenATP from different sources,for example,from different typesof microorganisms,such as bacteria and fungi.1.6 The values stated in SI are to be regarded as s
6、tandard.1.7 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.Referenc
7、ed Documents2.1 ASTM Standards:2D1129 Terminology Relating to WaterD4012 Test Method forAdenosine Triphosphate(ATP)Con-tent of Microorganisms in WaterD4840 Guide for Sample Chain-of-Custody ProceduresD6161 Terminology Used for Microfiltration,Ultrafiltration,Nanofiltration and Reverse Osmosis Membra
8、ne ProcessesE177 Practice for Use of the Terms Precision and Bias inASTM Test MethodsE691 Practice for Conducting an Interlaboratory Study toDetermine the Precision of a Test MethodE1326 Guide for Evaluating Non-culture MicrobiologicalTests Used for Enumerating BacteriaE1497 Practice for Selection a
9、nd Safe Use of Water-Miscible and Straight Oil Metal Removal FluidsE2523 Terminology for Metalworking Fluids and Opera-tions2.2 Government Standards:329 CFR 1910.1000 Occupational Safety and Health Stan-dards;Air contaminants29 CFR 1910.1450 Occupational Exposure to HazardousChemicals in Laboratorie
10、s3.Terminology3.1 Definitions:For definition of terms used in this method,refer to Terminology standards D1129,D6161,and E2523.3.2 adenosine triphosphate(ATP),na molecule com-prised of a purine and three phosphate groups that serves as theprimary energy transport molecule in all biological cells.3.3
11、 adenosine monophosphate(AMP),nthe moleculeformed by the removal of two molecules of phosphate(onepyrophosphate molecule)from ATP.3.4 aseptic,adjsterile,free from viable microbial con-tamination.3.5 bioluminescence,nthe production and emission oflight by a living organism as the result of a chemical
12、 reactionduring which chemical energy is converted to light energy.3.6 biomass,nany matter which is or was a livingorganism or excreted from a microorganism(D6161).1This test method is under the jurisdiction of ASTM Committee E34 onOccupational Health and Safety and is the direct responsibility of S
13、ubcommitteeE34.50 on Health and Safety Standards for Metal Working Fluids.Current edition approved Aug.1,2011.Published August 2011.Originallyapproved in 2009.Last previous edition approved in 2009 as E2694-09.DOI:10.1520/E2694-11.2For referenced ASTM standards,visit the ASTM website,www.astm.org,or
14、contact ASTM Customer Service at serviceastm.org.For Annual Book of ASTMStandards volume information,refer to the standards Document Summary page onthe ASTM website.3Available from U.S.Government Printing Office Superintendent of Documents,732 N.Capitol St.,NW,Mail Stop:SDE,Washington,DC 20401,http:
15、/www.access.gpo.gov.*A Summary of Changes section appears at the end of this standardCopyright ASTM International,100 Barr Harbor Drive,PO Box C700,West Conshohocken,PA 19428-2959.United States1 3.7 culturable,adjmicroorganisms that proliferate as in-dicated by the formation of colonies on solid gro
16、wth media orthe development of turbidity in liquid growth media underspecific growth conditions.3.8 Luciferase,na general term for a class of enzymes thatcatalyze bioluminescent reactions.3.9 Luciferin,na general term for a class of light-emittingbiological pigments found in organisms capable of biolumi-nescence.3.10 luminometer,nan instrument capable of measuringlight emitted as a result of non-thermal excitation.3.11 relative light unit(RLU),nan instrument-specificunit of measurement reflectin
