ASTM D7687-17

Standard Test Method for Measurement of Cellular Adenosine Triphosphate in Fuel and Fuel-associated Water With Sample Concentration by Filtration

SIGNIFICANCE AND USE
5.1 This test method measures the concentration of cellular-ATP present in the sample. ATP is a constituent of all living cells, including bacteria and fungi. Consequently, the presence of cellular-ATP is an indicator of total metabolically active microbial contamination in fuels. ATP is not associated with matter of non-biological origin.  
5.2 This test method is similar to Test Method E2694 except for the volumes sampled.  
5.3 This test method differs from Test Method D4012 in that it utilizes filtration and wash steps designed to eliminate interferences that have historically rendered ATP testing unusable with complex organic fluids such as fuel and fuel-associated water.  
5.4 This test method differs from Test Method D7463 in several regards:  
5.4.1 Test Method D7463 reports relative light units (RLU). Consistent with Test Methods D4012 and E2694, this test method reports ATP concentration.  
5.4.2 This test method detects only cellular-ATP and it can be used to detect cellular-ATP in fuels and fuel stocks from which small quantities of water do not separate readily (for example, ethanol blended gasoline containing ≥5 % v/v ethanol). Test Method D7463 cannot be used to recover ATP from fuels from which small quantities of water do not separate readily (for example, ethanol blended gasoline containing ≥5 % v/v ethanol).  
5.4.3 This test method measures cellular-ATP in a single measurement (as pg ATP/mL). Test Method D7463 detects total ATP (as RLU) and extra-cellular ATP (as RLU) using two separate analyses and permits computation of cellular-ATP (as RLU) as the difference between total and extracellular ATP.  
5.4.4 Test Method D7463 suggests a nominal 500 mL fuel sample volume. This test method suggests a nominal 20 mL fuel sample.  
5.5 This test method can be used with all fuels specified in Specifications D396, D975, D1655, D2069, D2880, D3699, D6751, and D7467 and other fuels with nominal viscosities ≤75 cSt at 20° ± 2°.  
5.6 The ATP test pro...
SCOPE
1.1 This test method covers a protocol for capturing, extracting and quantifying the cellular adenosine triphosphate (cellular-ATP) content associated with microorganisms found in fuels and fuel-associated water.  
1.2 The ATP is measured using a bioluminescence enzyme assay, whereby light is generated in amounts proportional to the concentration of cellular-ATP in the samples. The light is produced and measured quantitatively as relative light units (RLU) which are converted by comparison with an ATP standard, computation to pg ATP/mL and optional further transformation to Log10[pg ATP/mL].  
1.3 This test method is equally suitable for use as a laboratory or portable method.  
1.4 This test method is limited to fuels with a nominal viscosity ≤75 cSt at test temperature.  
1.5 This test method detects ATP concentrations in the range of 5.0 pg ATP/mL (≈0.699 log10[pg ATP/mL]) to 100 000 pg ATP/mL (≈5.000 log10[pg ATP/mL]) for 20 mL samples of fuel and 20 pg ATP/mL (≈1.301 log10[pg ATP/mL]) to 400 000 pg ATP/mL (≈5.602 log10[pg ATP/mL]) for 5 mL samples of fuel-associated water.
Note 1: These ranges were calculated with the formula for calculating sample ATP in pg/mL provided in 12.1 based on the minimum recommended RLU for a 1 ng/mL ATP standard when using the reagents specified in Section 7 and the luminometer specified in 6.4 and corrected with a reagent-method blank as determined in Appendix X5.  
1.6 Providing interferences can be overcome, bioluminescence is a reliable and proven method for qualifying and quantifying ATP. This test method does not differentiate between ATP from different sources, for example: from different types of microorganisms, such as bacteria and fungi.  
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.8 This standard does not purport to address all of the safety concerns, if any, associated with its us...

General Information

• Status: Historical
• Publication Date: 30-Apr-2017
• ICS: 75.160.20 - Liquid fuels
• Technical Committee: D02 - Petroleum Products, Liquid Fuels, and Lubricants
• Drafting Committee: D02.14 - Stability, Cleanliness and Compatibility of Liquid Fuels

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: D7687 − 17
Standard Test Method for
Measurement of Cellular Adenosine Triphosphate in Fuel
and Fuel-associated Water With Sample Concentration by
1
Filtration
This standard is issued under the fixed designation D7687; 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* 1.7 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 covers a protocol for capturing, ex-
standard.
tracting and quantifying the cellular adenosine triphosphate
1.8 This standard does not purport to address all of the
(cellular-ATP) content associated with microorganisms found
safety concerns, if any, associated with its use. It is the
in fuels and fuel-associated water.
responsibility of the user of this standard to establish appro-
1.2 The ATP is measured using a bioluminescence enzyme
priate safety and health practices and determine the applica-
assay, whereby light is generated in amounts proportional to
bility of regulatory limitations prior to use.
the concentration of cellular-ATP in the samples. The light is
1.9 This international standard was developed in accor-
produced and measured quantitatively as relative light units
dance with internationally recognized principles on standard-
(RLU) which are converted by comparison with an ATP
ization established in the Decision on Principles for the
standard, computation to pg ATP/mL and optional further
Development of International Standards, Guides and Recom-
transformation to Log [pg ATP/mL].
10
mendations issued by the World Trade Organization Technical
1.3 This test method is equally suitable for use as a Barriers to Trade (TBT) Committee.
laboratory or portable method.
2. Referenced Documents
1.4 This test method is limited to fuels with a nominal
2
2.1 ASTM Standards:
viscosity ≤75 cSt at test temperature.
D396 Specification for Fuel Oils
1.5 ThistestmethoddetectsATPconcentrationsintherange
D975 Specification for Diesel Fuel Oils
of 5.0 pg ATP/mL (≈0.699 log [pg ATP/mL]) to 100 000 pg
10
D1129 Terminology Relating to Water
ATP/mL(≈5.000log [pgATP/mL])for20 mLsamplesoffuel
10
D1655 Specification for Aviation Turbine Fuels
and 20 pg ATP/mL (≈1.301 log [pg ATP/mL]) to 400 000 pg
3
10
D2069 Specification for Marine Fuels (Withdrawn 2003)
ATP/mL (≈5.602 log [pg ATP/mL]) for 5 mL samples of
10
D2880 Specification for Gas Turbine Fuel Oils
fuel-associated water.
D3699 Specification for Kerosine
NOTE 1—These ranges were calculated with the formula for calculating D4012 Test Method forAdenosineTriphosphate (ATP) Con-
sample ATP in pg/mL provided in 12.1 based on the minimum recom-
tent of Microorganisms in Water
mended RLU for a 1 ng⁄mL ATP standard when using the reagents
D4175 Terminology Relating to Petroleum Products, Liquid
specified in Section 7 and the luminometer specified in 6.4 and corrected
Fuels, and Lubricants
with a reagent-method blank as determined in Appendix X5.
D6161 Terminology Used for Microfiltration, Ultrafiltration,
1.6 Providing interferences can be overcome, biolumines-
Nanofiltration and Reverse Osmosis Membrane Processes
cence is a reliable and proven method for qualifying and
D6300 Practice for Determination of Precision and Bias
quantifying ATP. This test method does not differentiate
Data for Use in Test Methods for Petroleum Products and
between ATP from different sources, for example: from differ-
Lubricants
ent types of microorganisms, such as bacteria and fungi.
D6751 Specification for Biodiesel Fuel Blend Stock (B100)
1 2
This test method is under the jurisdiction of ASTM Committee D02 on For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Petroleum Products, Liquid Fuels, and Lubricants and is the direct responsibility of contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
Subcommittee D02.14 on Stability and Cleanliness of Liquid Fuels. Standards volume information, refer to the standard’s Document Summary page on
Current edition approved May 1, 2017. Published June 2017. Originally the ASTM website.
3
approved in 2011. Last previous edition approved in 2011 as D7687 – 11. The last approved version of this historical standard is referenced on
DOI:10.1520 ⁄D7687-17. 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

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D7687 − 17
for Middle Distillate Fuels 3.1.10.1 Discussion—Extra-cellular ATP is ATP that has
D7463 T
...

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: D7687 − 11 D7687 − 17
Standard Test Method for
Measurement of Cellular Adenosine Triphosphate in Fuel,
Fuel/Water Mixtures, and Fuel-Associated Water withFuel
and Fuel-associated Water With Sample Concentration by
1
Filtration
This standard is issued under the fixed designation D7687; 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 covers a protocol for capturing, extracting and quantifying the cellular adenosine triphosphate
(cellular-ATP) content associated with microorganisms found in fuels, fuel/water mixtures fuels and fuel-associated water.
1.2 The ATP is measured using a bioluminescence enzyme assay, whereby light is generated in amounts proportional to the
concentration of cellular-ATP in the samples. The light is produced and measured quantitatively as relative light units (RLU) which
are converted by comparison with an ATP standard, computation to pg ATP/mL and optional further transformation to Log [pg
10
ATP/mL].
1.3 This test method is equally suitable for use as a laboratory or portable method.
1.4 This test method is limited to fuels with a nominal viscosity ≤ 75cSt ≤75 cSt at test temperature.
1.5 This test method detects ATP concentrations in the range of 5.0 pg ATP/mL (0.699(≈0.699 log [pg ATP/mL]) to
10
100 000100 000 pg ATP/mL (5.000(≈5.000 log [pg ATP/mL]) for 20 mL 20 mL samples of fuel or fuel/water mixtures, and 20
10
pg ATP/mL (1.301and 20 pg ATP/mL (≈1.301 log [pg ATP/mL]) to 400 000400 000 pg ATP/mL (5.602(≈5.602 log [pg
10 10
ATP/mL]) for 5 mL samples of fuel-associated water.
NOTE 1—These ranges were calculated with the formula for calculating sample ATP in pg/mL provided in 12.1 based on the minimum recommended
RLU for a 11 ng ng ATP/mL ⁄mL ATP standard when using the reagents specified in Section 7 and the luminometer specified in 6.4 and corrected with
a reagent-method blank as determined in Appendix X5.
1.6 Providing interferences can be overcome, bioluminescence is a reliable and proven method for qualifying and quantifying
ATP. This test method does not differentiate between ATP from different sources, for example: from different types of
microorganisms, such as bacteria and fungi.
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.8 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.
1.9 This 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
2
2.1 ASTM Standards:
D396 Specification for Fuel Oils
D975 Specification for Diesel Fuel Oils
D1129 Terminology Relating to 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 April 1, 2011May 1, 2017. Published April 2011June 2017. DOI:10.1520/D7687–11.Originally approved in 2011. Last previous edition approved
in 2011 as D7687 – 11. DOI:10.1520 ⁄D7687-17.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
*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 ----------------------
D7687 − 17
D1655 Specification for Aviation Turbine Fuels
3
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
D4175 Termi
...