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Standard Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in Fuel, Fuel/Water Mixtures, and Fuel Associated WaterTranslate name
STANDARD published on 1.4.2021
Designation standards: ASTM D7463-21
Publication date standards: 1.4.2021
The number of pages: 8
Approximate weight : 24 g (0.05 lbs)
Country: American technical standard
Category: Technical standards ASTM
adenosine triphosphate, assay, ATP, ATP Bioluminescence Assay, bacteria, biocontamination, biodeterioration, biomass, capture solution, cellular, extracellular, fuel, fungi, hydrophilic particles, luciferin, luciferase, luminometer, microbe, microbiology, Relative Light Unit (RLU),, ICS Number Code 07.100.99 (Other standards related to microbiology),75.160.20 (Liquid fuels)
|Significance and Use|
5.1This test method measures the concentration of ATP present in the sample. ATP is a constituent of all living cells including bacteria and fungi. Consequently, the presence of ATP is a reliable indicator of microbial contamination in fuel systems. ATP is not associated with matter of non-biological origin.
5.2This test method differs from Test Method as follows:
5.2.1By providing for the rapid determination of ATP present in a fuel (petroleum) sample, a fuel and water mixture sample, fuel-associated bottom water sample, and extracellular ATP freely available in the fuel or aqueous sample matrix;
5.2.2By providing for a method to capture, extract, and quantify ATP using self-contained test device and luminometer;
5.2.3By providing a method of quantifying ATP present in fuel or water matrices in generally less than 10 min; and
5.2.4By providing for the rapid separation of the ATP from chemical interferences that have previously prevented the use of ATP determinations in complex fluids containing hydrocarbons and other organic molecules.
5.3This test method does not require the use of hazardous materials and does not generate biohazard waste.
5.4This test method can be used to estimate viable microbial biomass, to evaluate the efficacy of antimicrobial pesticides, and to monitor microbial contamination in fuel storage and distribution systems.
1.1This 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:
1.1.1Microorganisms and hydrophilic particles found in liquid fuels as described in , or
1.1.2Microorganisms and hydrophilic particles found in mixture of fuel and associated bottom water or just associated bottom water.
1.1.3ATP detected by this bioluminescence test can be derived from cellular ATP, extra-cellular ATP, or some combination of both.
1.1.4Cellular and extra-cellular ATP utilized to perform ATP bioluminescence are captured and concentrated from a fuel system sample into an aqueous test specimen (that is, sub-sample) for testing. For example, for a fuel system sample that does not contain any visible fuel associated bottom water, the aqueous test specimen is the capture solution itself described in . For fuel system samples that are a mixture of fuel and associated bottom water (that is, free water), the test specimen is an aliquant of the capture solution and associated bottom water.
1.2The ATP is measured using a patented bioluminescence enzyme assay, whereby light is generated in amounts proportional to the concentration of ATP in the sample. The light is produced and measured quantitatively using dedicated ATP test pens and a dedicated luminometer and reported in (instrument specific) Relative Light Units.
1.3This test method is equally suitable for use in the laboratory or field.
1.4Although bioluminescence is a reliable and proven technology, this method does not differentiate ATP from bacteria or fungi.
1.5For water or capture solution samples, the concentration range of ATP detectable by this test method is 1 1011 M to 3 108 M which is equivalent to 1 1014 moles/mL to 3 1011 moles/mL for water samples or capture solution. Assuming testing on fuel phase is performed on a 500mL volume of fuel the equivalent concentrations is fuel would be: 6 1011M to 2 1014 M.
1.6The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.6.1There is one exceptionRelative Light Unit (RLU) as defined in .
1.7This 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.
1.8This 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.
|2. Referenced Documents|
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