ASTM D7687-11
(Test Method)Standard Test Method for Measurement of Cellular Adenosine Triphosphate in Fuel, Fuel/Water Mixtures, and Fuel-Associated Water with Sample Concentration by Filtration
Standard Test Method for Measurement of Cellular Adenosine Triphosphate in Fuel, Fuel/Water Mixtures, and Fuel-Associated Water with Sample Concentration by Filtration
SIGNIFICANCE AND USE
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.
This test method is similar to Test Method E2694 except for the volumes sampled.
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, fuel/water mixtures and fuel-associated water.
This test method differs from Test Method D7463 in several regards:
Test Method D7463 reports relative light units (RLU). Consistent with Test Method D4012 and E2694, this test method reports ATP concentration.
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).
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.
Test Method D7463 suggests a nominal 500 mL fuel sample volume. This test method suggests a nominal 20 mL fuel sample.
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°.
The ATP test provides rapid test results that reflect...
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 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 ≤ 75cSt 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 or fuel/water mixtures, 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 ATP/mL 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 concer...
General Information
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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 − 11
Standard Test Method for
Measurement of Cellular Adenosine Triphosphate in Fuel,
Fuel/Water Mixtures, and Fuel-Associated Water with
1
Sample Concentration by 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 1.8 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
1.1 This test method covers a protocol for capturing, ex-
responsibility of the user of this standard to establish appro-
tracting and quantifying the cellular adenosine triphosphate
priate safety and health practices and determine the applica-
(cellular-ATP) content associated with microorganisms found
bility of regulatory limitations prior to use.
in fuels, fuel/water mixtures and fuel-associated water.
1.2 The ATP is measured using a bioluminescence enzyme
2. Referenced Documents
assay, whereby light is generated in amounts proportional to
2
2.1 ASTM Standards:
the concentration of cellular-ATP in the samples. The light is
D396 Specification for Fuel Oils
produced and measured quantitatively as relative light units
D975 Specification for Diesel Fuel Oils
(RLU) which are converted by comparison with an ATP
D1129 Terminology Relating to Water
standard, computation to pg ATP/mL and optional further
D1655 Specification for Aviation Turbine Fuels
transformation to Log [pg ATP/mL].
10
3
D2069 Specification for Marine Fuels (Withdrawn 2003)
1.3 This test method is equally suitable for use as a
D2880 Specification for Gas Turbine Fuel Oils
laboratory or portable method.
D3699 Specification for Kerosine
1.4 This test method is limited to fuels with a nominal
D4012 Test Method forAdenosineTriphosphate (ATP) Con-
viscosity ≤ 75cSt at test temperature.
tent of Microorganisms in Water
1.5 ThistestmethoddetectsATPconcentrationsintherange D4175 Terminology Relating to Petroleum, Petroleum
Products, and Lubricants
of 5.0 pg ATP/mL (0.699 log [pg ATP/mL]) to 100 000 pg
10
ATP/mL (5.000 log [pg ATP/mL]) for 20 mL samples of fuel D6161 Terminology Used for Microfiltration, Ultrafiltration,
10
or fuel/water mixtures, and 20 pg ATP/mL (1.301 log [pg Nanofiltration and Reverse Osmosis Membrane Processes
10
ATP/mL]) to 400 000 pg ATP/mL (5.602 log [pg ATP/mL]) D6300 Practice for Determination of Precision and Bias
10
for 5 mL samples of fuel-associated water. Data for Use in Test Methods for Petroleum Products and
Lubricants
NOTE 1—These ranges were calculated with the formula for calculating
D6751 Specification for Biodiesel Fuel Blend Stock (B100)
sample ATP in pg/mL provided in 12.1 based on the minimum recom-
mended RLU fora1ngATP/mL standard when using the reagents
for Middle Distillate Fuels
specified in Section 7 and the luminometer specified in 6.4 and corrected
D7463 Test Method forAdenosineTriphosphate (ATP) Con-
with a reagent-method blank as determined in Appendix X5.
tent of Microorganisms in Fuel, Fuel/Water Mixtures and
1.6 Providing interferences can be overcome, biolumines-
Fuel Associated Water
cence is a reliable and proven method for qualifying and
D7464 Practice for Manual Sampling of Liquid Fuels, As-
quantifying ATP. This test method does not differentiate
sociated Materials and Fuel System Components for
between ATP from different sources, for example: from differ-
Microbiological Testing
ent types of microorganisms, such as bacteria and fungi.
D7467 Specification for Diesel Fuel Oil, Biodiesel Blend
1.7 The values stated in SI units are to be regarded as (B6 to B20)
standard. No other units of measurement are included in this
standard.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
1
This test method is under the jurisdiction of ASTM Committee D02 on contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Petroleum Products and Lubricants and is the direct responsibility of Subcommittee Standards volume information, refer to the standard’s Document Summary page on
D02.14 on Stability and Cleanliness of Liquid Fuels. the ASTM website.
3
Current edition approved April 1, 2011. Published April 2011. DOI:10.1520/ The last approved version of this historical standard is referenced on
D7687–11. www.astm.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
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D7687 − 11
E2523 Terminology for Metalworking Fluids and Opera- 3.1.13 luminometer, n—instrument capable of measuring
tions light emitted as a result of nonthermal excita
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