ASTM D7463-14
(Test Method)Standard Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in Fuel, Fuel/Water Mixtures and Fuel Associated Water
Standard Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in Fuel, Fuel/Water Mixtures and Fuel Associated Water
SIGNIFICANCE AND USE
5.1 This 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.
5.2 This test method differs from Test Method D4012 as follows:
5.2.1 By 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;
5.2.2 By providing for a method to capture, extract and quantify ATP using self-contained test device and luminometer;
5.2.3 By providing a method of quantifying ATP present in fuel or water matrices in generally less than 10 min; and
5.2.4 By 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.
5.3 This test method does not require the use of hazardous materials and does not generate biohazard waste.
5.4 This 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.
SCOPE
1.1 This test method provides a protocol for capturing, extracting and quantifying the adenosine triphosphate (ATP) content associated with:
1.1.1 Microorganisms found in conventional liquid fuels with kinematic viscosities (at 40°C) of ≤ 8 mm2 · s–1 as described in Table X6.1,
1.1.2 Microorganisms found in fuel-associated bottom water, and
1.1.3 Extracellular (non-cellular) ATP present in the sample matrix.
1.2 The 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 pens2 and a dedicated luminometer2 and reported in (instrument specific) Relative Light Units.
1.3 This test method is equally suitable for use in the laboratory or field.
1.4 Although bioluminescence is a reliable and proven method for qualifying and quantifying ATP, this method does not differentiate between ATP from different sources, for example, from different types of microorganism such as bacteria or fungi.
1.5 For water or capture solution samples, the concentration range of ATP detectable by this test method is 1 × 10–11 M to 3 × 10–8 M which is equivalent to 1 × 10–14 moles/mL to 3 × 10–11 moles/mL for water samples or capture solution. Assuming testing on fuel phase is performed on a 500 mL volume of fuel the equivalent concentrations is fuel would be: 6 × 10–11 M to 2 × 10–14 M.
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.7 This 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 and health practices and determine the applicability of regulatory limitations prior to use.
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Designation: D7463 − 14
StandardTest Method for
Adenosine Triphosphate (ATP) Content of Microorganisms
1
in Fuel, Fuel/Water Mixtures and Fuel Associated Water
This standard is issued under the fixed designation D7463; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope* 1.6 The values stated in SI units are to be regarded as
standard. No other units of measurement are included in this
1.1 This test method provides a protocol for capturing,
standard.
extracting and quantifying the adenosine triphosphate (ATP)
1.7 This standard does not purport to address all of the
content associated with:
safety concerns, if any, associated with its use. It is the
1.1.1 Microorganisms found in conventional liquid fuels
2 –1
responsibility of the user of this standard to establish appro-
with kinematic viscosities (at 40°C) of ≤ 8mm ·s as
priate safety and health practices and determine the applica-
described in Table X6.1,
bility of regulatory limitations prior to use.
1.1.2 Microorganisms found in fuel-associated bottom
water, and
2. Referenced Documents
1.1.3 Extracellular (non-cellular)ATPpresent in the sample
3
matrix.
2.1 ASTM Standards:
D396Specification for Fuel Oils
1.2 TheATPis measured using a patented bioluminescence
D975Specification for Diesel Fuel Oils
enzyme assay, whereby light is generated in amounts propor-
D1129Terminology Relating to Water
tional to the concentration of ATP in the sample. The light is
D1655Specification for Aviation Turbine Fuels
producedandmeasuredquantitativelyusingdedicatedATPtest
4
2 2
D2069Specification for Marine Fuels (Withdrawn 2003)
pens and a dedicated luminometer and reported in (instru-
D2880Specification for Gas Turbine Fuel Oils
ment specific) Relative Light Units.
D3699Specification for Kerosine
1.3 This test method is equally suitable for use in the
D4012TestMethodforAdenosineTriphosphate(ATP)Con-
laboratory or field.
tent of Microorganisms in Water
1.4 Although bioluminescence is a reliable and proven
D4057Practice for Manual Sampling of Petroleum and
method for qualifying and quantifying ATP, this method does
Petroleum Products
not differentiate between ATP from different sources, for
D4175 Terminology Relating to Petroleum, Petroleum
example, from different types of microorganism such as
Products, and Lubricants
bacteria or fungi.
D6161TerminologyUsedforMicrofiltration,Ultrafiltration,
Nanofiltration and Reverse Osmosis Membrane Processes
1.5 Forwaterorcapturesolutionsamples,theconcentration
–11
D6469GuideforMicrobialContaminationinFuelsandFuel
range ofATP detectable by this test method is1×10 Mto
–8 –14
Systems
3×10 M which is equivalent to1×10 moles/mL to 3 ×
–11
D6751Specification for Biodiesel Fuel Blend Stock (B100)
10 moles/mLfor water samples or capture solution.Assum-
for Middle Distillate Fuels
ing testing on fuel phase is performed on a 500 mLvolume of
–11
fuel the equivalent concentrations is fuel would be:6×10
–14
3. Terminology
Mto2×10 M.
3.1 Definitions:
1 3.1.1 For definition of terms used in this test method, refer
This test method is under the jurisdiction of ASTM Committee D02 on
Petroleum Products, Liquid Fuels, and Lubricants and is the direct responsibility of to Terminologies D1129 and D4175, and Guide D6469.
Subcommittee D02.14 on Stability and Cleanliness of Liquid Fuels.
Current edition approved May 1, 2014. Published May 2014. Originally
approved in 2008. Last previous edition approved in 2008 as D7463–08. DOI:
3
10.1520/D7463-14. For referenced ASTM standards, visit the ASTM website, www.astm.org, or
2
The sole source of supply of the apparatus known to the committee at this time contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
is Merck KGaA, 64271 Darmstadt, Germany. If you are aware of alternative Standards volume information, refer to the standard’s Document Summary page on
suppliers, please provide this information to ASTM International Headquarters. the ASTM website.
4
Your comments will receive careful consideration at a meeting of the responsible The last approved version of this historical standard is referenced on
1
technical committee, which you may attend. www.astm.org.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D7463 − 14
3.1.2 adenosine triphosphate, n—molecule comprised of a 3.1.17 viable microbial biomass, n—metabolically active
purine and three phosphate groups, that serves as the primary (living) micro-orga
...
This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: D7463 − 08 D7463 − 14
Standard Test Method for
Adenosine Triphosphate (ATP) Content of Microorganisms
1
in Fuel, Fuel/Water Mixtures and Fuel Associated Water
This standard is issued under the fixed designation D7463; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope Scope*
1.1 This test method provides a protocol for capturing, extracting and quantifying the adenosine triphosphate (ATP) content
associated with:
2 –1
1.1.1 Microorganisms found in conventional liquid fuels with kinematic viscosities (at 40°C) of ≤ 8 mm · s as described in
Table X6.1,
1.1.2 Microorganisms found in fuel-associated bottom water, and
1.1.3 Extracellular (non-cellular) ATP present in the sample matrix.
1.2 The ATP is measured using a patented bioluminescence enzyme assay, whereby light is generated in amounts proportional
2
to the concentration of ATP in the sample. The light is produced and measured quantitatively using dedicated ATP test pens and
2
a dedicated luminometer and reported in (instrument specific) Relative Light Units.
1.3 This test method is equally suitable for use in the laboratory or field.
1.4 Although bioluminescence is a reliable and proven method for qualifying and quantifying ATP, this method does not
differentiate between ATP from different sources, for example, from different types of microorganism such as bacteria or fungi.
–11
1.5 For water or capture solution samples, the concentration range of ATP detectable by this test method is 1 × 10 M to 3
–8 –14 –11
× 10 M which is equivalent to 1 × 10 moles/mL to 3 × 10 moles/mL for water samples or capture solution. Assuming testing
–11 –14
on fuel phase is performed on a 500 mL volume of fuel the equivalent concentrations is fuel would be: 6 × 10 M to 2 × 10
M.
1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.7 This 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 and health practices and determine the applicability of regulatory
limitations prior to use.
2. Referenced Documents
3
2.1 ASTM Standards:
D396 Specification for Fuel Oils
D975 Specification for Diesel Fuel Oils
D1129 Terminology Relating to Water
D1655 Specification for Aviation Turbine Fuels
4
D2069 Specification for Marine Fuels (Withdrawn 2003)
D2880 Specification for Gas Turbine Fuel Oils
D3699 Specification for Kerosine
D4012 Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in Water
1
This test method is under the jurisdiction of ASTM Committee D02 on Petroleum Products Products, Liquid Fuels, and Lubricants and is the direct responsibility of
Subcommittee D02.14 on Stability and Cleanliness of Liquid Fuels.
Current edition approved Dec. 1, 2008May 1, 2014. Published February 2009May 2014. Originally approved in 2008. Last previous edition approved in 2008 as
D7463 – 08. DOI: 10.1520/D7463-08.10.1520/D7463-14.
2
The sole source of supply of the apparatus known to the committee at this time is Merck KGaA, 64271 Darmstadt, Germany. If you are aware of alternative suppliers,
1
please provide this information to ASTM International Headquarters. Your comments will receive careful consideration at a meeting of the responsible technical committee,
which you may attend.
3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
4
The last approved version of this historical standard is referenced on www.astm.org.
*A Summary of Changes section appears at the end of this standard
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
---------------------- Page: 1 ----------------------
D7463 − 14
D4057 Practice for Manual Sampling of Petroleum and Petroleum Products
D4175 Terminology Relating to Petroleum, Petroleum Products, and Lubricants
D6161 Terminology Used for Microfiltration, Ultrafiltration, Nanofiltration and Reverse Osmosis Membrane Processes
D6469 Guide for Microbial Contamination in Fuels and Fuel Systems
D6751 Speci
...
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