ASTM E2694-21
(Test Method)Standard Test Method for Measurement of Adenosine Triphosphate in Water-Miscible Metalworking Fluids
Standard Test Method for Measurement of Adenosine Triphosphate in Water-Miscible Metalworking Fluids
SIGNIFICANCE AND USE
5.1 This 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 an indicator of total microbial contamination in metalworking fluids. ATP is not associated with matter of non-biological origin.
5.2 Test Method D4012 validated ATP as a surrogate for culturable bacterial data (Guide E1326).
5.3 This method differs from Test Method D4012 in that it eliminates interferences that have historically rendered ATP testing unusable with complex organic fluids such as MWFs.
5.4 The ATP test provides rapid test results that reflect the total bioburden in the sample. It thereby reduces the delay between test initiation and data capture, from the 36 h to 48 h (or longer) required for culturable colonies to become visible, to approximately 5 min.
5.5 Although ATP data generally covary with culture data in MWF,4 different factors affect ATP concentration than those that affect culturability.
5.5.1 Culturability is affected primarily by the ability of captured microbes to proliferate on the growth medium provided, under specific growth conditions. It has been estimated that less than 1 % of the species present in an environmental sample will form colonies under any given set of growth conditions.5
5.5.2 ATP concentration is affected by: the microbial species present, the physiological states of those species, and the total bioburden (see Appendix X1).
5.5.2.1 One example of the species effect is that the amount of ATP per cell is substantially greater for fungi than bacteria.
5.5.2.2 Within a species, cells that are more metabolically active will have more ATP per cell than dormant cells.
5.5.2.3 The greater the total bioburden, the greater the ATP concentration in a sample.
5.5.3 The possibility exists that the rinse step (11.15) may not eliminate all chemical substances that can interfere with the bioluminescence reaction (11.39).
5.5.3...
SCOPE
1.1 This test method provides a protocol for capturing, extracting, and quantifying the adenosine triphosphate (ATP) content associated with microorganisms found in water-miscible metalworking fluids (MWFs).
1.2 The ATP is measured using a bioluminescence enzyme assay, whereby light is generated in amounts proportional to the concentration of ATP in the samples. The light is produced and measured quantitatively as relative light units (RLUs) which are converted by comparison with an ATP standard and computation to pg ATP/mL.
1.3 This test method is equally suitable for use in the laboratory or field.
1.4 The test method detects ATP concentrations in the range of 4.0 pg ATP/mL to 400 000 pg ATP/mL.
1.5 Providing interferences can be overcome, bioluminescence is a reliable and proven method for qualifying and quantifying ATP. The method does not differentiate between ATP from different sources, for example, from different types of microorganisms, such as bacteria and fungi.
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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.
1.8 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.
General Information
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Standards Content (Sample)
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.
Designation: E2694 − 21 An American National Standard
Standard Test Method for
Measurement of Adenosine Triphosphate in Water-Miscible
1
Metalworking Fluids
This standard is issued under the fixed designation E2694; 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* 2. Referenced Documents
2
1.1 This test method provides a protocol for capturing, 2.1 ASTM Standards:
extracting, and quantifying the adenosine triphosphate (ATP) D1129Terminology Relating to Water
content associated with microorganisms found in water- D4012TestMethodforAdenosineTriphosphate(ATP)Con-
miscible metalworking fluids (MWFs). tent of Microorganisms in Water
D4840Guide for Sample Chain-of-Custody Procedures
1.2 The ATP is measured using a bioluminescence enzyme
D6161TerminologyUsedforMicrofiltration,Ultrafiltration,
assay, whereby light is generated in amounts proportional to
Nanofiltration,andReverseOsmosisMembraneProcesses
the concentration ofATPin the samples. The light is produced
E177Practice for Use of the Terms Precision and Bias in
and measured quantitatively as relative light units (RLUs)
ASTM Test Methods
which are converted by comparison with anATP standard and
E691Practice for Conducting an Interlaboratory Study to
computation to pg ATP/mL.
Determine the Precision of a Test Method
1.3 This test method is equally suitable for use in the
E1326Guide for Evaluating Non-culture Microbiological
laboratory or field.
Tests
1.4 ThetestmethoddetectsATPconcentrationsintherange E1497 Practice for Selection and Safe Use of Water-
Miscible and Straight Oil Metal Removal Fluids
of 4.0 pg ATP/mL to 400 000 pg ATP/mL.
E2523Terminology for Metalworking Fluids and Opera-
1.5 Providing interferences can be overcome, biolumines-
tions
cence is a reliable and proven method for qualifying and
3
2.2 Government Standards:
quantifying ATP. The method does not differentiate between
29 CFR 1910.1000Air Contaminants
ATP from different sources, for example, from different types
29 CFR 1910.1450Occupational Exposure to Hazardous
of microorganisms, such as bacteria and fungi.
Chemicals in Laboratories
1.6 The values stated in SI units are to be regarded as
standard. No other units of measurement are included in this 3. Terminology
standard.
3.1 Definitions:
1.7 This standard does not purport to address all of the
3.1.1 For definitions of terms used in this test method, refer
safety concerns, if any, associated with its use. It is the
to Terminologies D1129, D6161, and E2523.
responsibility of the user of this standard to establish appro-
3.1.2 adenosine monophosphate (AMP), n—the molecule
priate safety, health, and environmental practices and deter-
formed by the removal of two molecules of phosphate (one
mine the applicability of regulatory limitations prior to use. pyrophosphate molecule) from ATP.
1.8 This international standard was developed in accor-
3.1.3 adenosine triphosphate (ATP), n—a molecule com-
dance with internationally recognized principles on standard-
prisedofapurineandthreephosphategroupsthatservesasthe
ization established in the Decision on Principles for the
primary energy transport molecule in all biological cells.
Development of International Standards, Guides and Recom-
3.1.4 aseptic, adj—sterile, free from viable microbial con-
mendations issued by the World Trade Organization Technical
tamination.
Barriers to Trade (TBT) Committee.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
1
This test method is under the jurisdiction of ASTM Committee E34 on contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Occupational Health and Safety and is the direct responsibility of Subcommittee Standards volume information, refer to the standard’s Document Summary page on
E34.50 on Health and Safety Standards for Metal Working Fluids. the ASTM website.
3
Current edition approved Nov. 1, 2021. Published November 2021. Originally AvailablefromU.S.GovernmentPrintingOfficeSuperintendentofDocuments,
approved in 2009. Last previous edition approved in 2016 as E2694–16. 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
DOI:10.1520/E2694-21. www.access.gpo.gov.
*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 ----------------------
E2694 − 21
3.1.5 bioluminesce
...
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: E2694 − 16 E2694 − 21 An American National Standard
Standard Test Method for
Measurement of Adenosine Triphosphate in Water-Miscible
1
Metalworking Fluids
This standard is issued under the fixed designation E2694; 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*
1.1 This test method provides a protocol for capturing, extracting, and quantifying the adenosine triphosphate (ATP) content
associated with microorganisms found in water-miscible metalworking fluids (MWF).(MWFs).
1.2 The ATP is measured using a bioluminescence enzyme assay, whereby light is generated in amounts proportional to the
concentration of ATP in the samples. The light is produced and measured quantitatively as relative light units (RLU)(RLUs) which
are converted by comparison with an ATP standard and computation to pg ATP/mL.
1.3 This test method is equally suitable for use in the laboratory or field.
1.4 The test method detects ATP concentrations in the range of 4.0 pg ATP/mL to 400 000 pg ATP/mL.
1.5 Providing interferences can be overcome, bioluminescence is a reliable and proven method for qualifying and quantifying ATP.
The method does not differentiate between ATP from different sources, for example, from different types of microorganisms, such
as bacteria and fungi.
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 safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.8 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:
D1129 Terminology Relating to Water
D4012 Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in Water
1
This test method is under the jurisdiction of ASTM Committee E34 on Occupational Health and Safety and is the direct responsibility of Subcommittee E34.50 on Health
and Safety Standards for Metal Working Fluids.
Current edition approved Oct. 1, 2016Nov. 1, 2021. Published October 2016November 2021. Originally approved in 2009. Last previous edition approved in 20112016
as E2694 - 11.E2694 – 16. DOI:10.1520/E2694-16. DOI:10.1520/E2694-21.
2
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.
*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 ----------------------
E2694 − 21
D4840 Guide for Sample Chain-of-Custody Procedures
D6161 Terminology Used for Microfiltration, Ultrafiltration, Nanofiltration, and Reverse Osmosis Membrane Processes
E177 Practice for Use of the Terms Precision and Bias in ASTM Test Methods
E691 Practice for Conducting an Interlaboratory Study to Determine the Precision of a Test Method
E1326 Guide for Evaluating Non-culture Microbiological Tests
E1497 Practice for Selection and Safe Use of Water-Miscible and Straight Oil Metal Removal Fluids
E2523 Terminology for Metalworking Fluids and Operations
3
2.2 Government Standards:
29 CFR 1910.1000 Occupational Safety and Health Standards; Air contaminantsAir Contaminants
29 CFR 1910.1450 Occupational Exposure to Hazardous Chemicals in Laboratories
3. Terminology
3.1 Definitions: For definition of terms used in this method, refer to Terminology standards D1129, D6161, and E2523.
3.1 Definitions:
3.1.1 For definitions of terms used in this test method, refer to Terminologies D1129, D6161, and E2523.
3.1.2 adenosine monophosphate (AMP), n—the molecule formed by the removal of two molecules of phosphate (one
pyrophosphate molecule) from ATP.
3.
...
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