ASTM_D_7463_-_15.pdf

Designation:D746315Standard Test Method forAdenosine Triphosphate(ATP)Content of Microorganismsin Fuel,Fuel/Water Mixtures,and Fuel Associated Water1This standard is issued under the fixed designation D7463;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()indicates an editorial change since the last revision or reapproval.1.Scope*1.1 This test method provides a protocol for capturing,concentrating,and testing the adenosine triphosphate(ATP)present in a fuel system sub-sample(that is,test specimen)associated with:1.1.1 Microorganisms and hydrophilic particles found inliquid fuels as described in Table X6.1,or1.1.2 Microorganisms and hydrophilic particles found inmixture of fuel and associated bottom water or just associatedbottom water.1.1.3 ATP detected by this bioluminescence test can bederived from cellular ATP,extra-cellular ATP,or some combi-nation of both.1.1.4 Cellular and extra-cellular ATP utilized to performATP bioluminescence are captured and concentrated from afuel system sample into an aqueous test specimen(that is,sub-sample)for testing.For example,for a fuel system samplethat does not contain any visible fuel associated bottom water,the aqueous test specimen is the capture solution itself de-scribed in 8.2.1.1.For fuel system samples that are a mixtureof fuel and associated bottom water(that is,free water),the testspecimen is an aliquant of the capture solution and associatedbottom water.1.2 The ATP is measured using a patented bioluminescenceenzyme assay,whereby light is generated in amounts propor-tional to the concentration of ATP in the sample.The light isproduced and measured quantitatively using dedicatedATPtestpens2and a dedicated luminometer2and reported in(instru-ment specific)Relative Light Units.1.3 This test method is equally suitable for use in thelaboratory or field.1.4 Although bioluminescence is a reliable and proventechnology,this method does not differentiate ATP frombacteria or fungi.1.5 For water or capture solution samples,the concentrationrange of ATP detectable by this test method is 1 1011M to3 108M which is equivalent to 1 1014moles/mL to 3 1011moles/mL for water samples or capture solution.Assum-ing testing on fuel phase is performed on a 500 mL volume offuel the equivalent concentrations is fuel would be:6 1011M to 2 1014M.1.6 The values stated in SI units are to be regarded asstandard.No other units of measurement are included in thisstandard.1.6.1 There is one exceptionRelative Light Unit(RLU)asdefined in 3.1.19.1.7 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.Referenced Documents2.1 ASTM Standards:3D396 Specification for Fuel OilsD975 Specification for Diesel Fuel OilsD1655 Specification for Aviation Turbine FuelsD2880 Specification for Gas Turbine Fuel OilsD4012 Test Method forAdenosine Triphosphate(ATP)Con-tent of Microorganisms in WaterD4175 Terminology Relating to Petroleum,PetroleumProducts,and LubricantsD6300 Practice for Determination of Precision and BiasData for Use in Test Methods for Petroleum Products andLubricants1This test method is under the jurisdiction of ASTM Committee D02 onPetroleum Products,Liquid Fuels,and Lubricants and is the direct responsibility ofSubcommittee D02.14 on Stability and Cleanliness of Liquid Fuels.Current edition approved Dec.1,2015.Published January 2016.Originallyapproved in 2008.Last previous edition approved in 2014 as D7463 14a.DOI:10.1520/D7463-15.2The sole source of supply,repair,recertification,and technical support of theapparatus or test pen known to the committee at this time is Merck KGaA,64271Darmstadt,Germany(Worldwide)or Fuel Quality Services,Inc.,4584 Cantrell Rd.,Flowery Branch,GA 30542(USA).If you are aware of alternative suppliers,pleaseprovide this information to ASTM International Headquarters.Your comments willreceive careful consideration at a meeting of the responsible technical committee,1which you may attend.3For referenced ASTM standards,visit the ASTM website,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.*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 D7464 Practice for Manual Sampling of Liquid Fuels,As-sociated Materials and Fuel System Components forMicrobiological TestingD7467 Specification for Diesel Fuel Oil,Biodiesel Blend(B6 to B20)3.Terminology3.1 Definitions:3.1.1 For definition of terms used in this test method,referto Terminol