ASTM E2694-11
(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
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.
Method D4012 validated ATP as a surrogate for culturable bacterial data (Guide E1326).
This method differs from Method D4012 in that it eliminates interferences that have historically rendered ATP testing unusable with complex organic fluids such as MWF.
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 five minutes.
Although ATP data generally covary with culture data in MWF , different factors affect ATP concentration than those that affect culturability.
Culturability is affected primarily by the ability of captured microbes to proliferate on the growth medium provided, under specific growth conditions. It have been estimated that less than 1 % of the species present in an environmental sample will form colonies under any given set of growth conditions.
ATP concentration is affected by: the microbial species present, the physiological states of those species, and the total bioburden (See Appendix X1).
One example of the species effect is that the amount of ATP per cell is substantially greater for fungi than bacteria.
Within a species, cells that are more metabolically active will have more ATP per cell than dormant cells.
The greater the total bioburden, the greater the ATP concentration in a sample.
The possibility exists that the rinse step (11.15) may not eliminate all chemical substances that can interfere with the bioluminescence reaction (11.39).
The presence of any such interferences can be evaluated by performing a stand...
SCOPE
1.1 The method provides a protocol for capturing, extracting and quantifying the adenosine triphosphate (ATP) content associated with microorganisms found in water-miscible metalworking fluids (MWF).
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) which are converted by comparison with an ATP standard and computation to pg ATP/mL.
1.3 This method is equally suitable for use in the laboratory or field.
1.4 The 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 are to be regarded as 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: E2694 − 11 AnAmerican 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* D4012 Test Method forAdenosineTriphosphate (ATP) Con-
tent of Microorganisms in Water
1.1 The method provides a protocol for capturing, extract-
D4840 Guide for Sample Chain-of-Custody Procedures
ing and quantifying the adenosine triphosphate (ATP) content
D6161 Terminology Used for Microfiltration, Ultrafiltration,
associated with microorganisms found in water-miscible met-
Nanofiltration and Reverse Osmosis Membrane Processes
alworking fluids (MWF).
E177 Practice for Use of the Terms Precision and Bias in
1.2 The ATP is measured using a bioluminescence enzyme
ASTM Test Methods
assay, whereby light is generated in amounts proportional to
E691 Practice for Conducting an Interlaboratory Study to
the concentration ofATP in the samples. The light is produced
Determine the Precision of a Test Method
and measured quantitatively as relative light units (RLU)
E1326 Guide for Evaluating Non-culture Microbiological
which are converted by comparison with an ATP standard and
Tests Used for Enumerating Bacteria
computation to pg ATP/mL.
E1497 Practice for Selection and Safe Use of Water-
1.3 This method is equally suitable for use in the laboratory Miscible and Straight Oil Metal Removal Fluids
E2523 Terminology for Metalworking Fluids and Opera-
or field.
tions
1.4 The method detects ATP concentrations in the range of
3
2.2 Government Standards:
4.0 pg ATP/mL to 400,000 pg ATP/mL.
29 CFR 1910.1000 Occupational Safety and Health Stan-
1.5 Providing interferences can be overcome, biolumines-
dards; Air contaminants
cence is a reliable and proven method for qualifying and
29 CFR 1910.1450 Occupational Exposure to Hazardous
quantifying ATP. The method does not differentiate between
Chemicals in Laboratories
ATP from different sources, for example, from different types
of microorganisms, such as bacteria and fungi.
3. Terminology
1.6 The values stated in SI are to be regarded as standard.
3.1 Definitions: For definition of terms used in this method,
1.7 This standard does not purport to address all of the refer to Terminology standards D1129, D6161, and E2523.
safety concerns, if any, associated with its use. It is the
3.2 adenosine triphosphate (ATP), n—a molecule com-
responsibility of the user of this standard to establish appro-
prised of a purine and three phosphate groups that serves as the
priate safety and health practices and determine the applica-
primary energy transport molecule in all biological cells.
bility of regulatory limitations prior to use.
3.3 adenosine monophosphate (AMP), n—the molecule
formed by the removal of two molecules of phosphate (one
2. Referenced Documents
pyrophosphate molecule) from ATP.
2
2.1 ASTM Standards:
3.4 aseptic, adj—sterile, free from viable microbial con-
D1129 Terminology Relating to Water
tamination.
3.5 bioluminescence, n—the production and emission of
light by a living organism as the result of a chemical reaction
1
This test method is under the jurisdiction of ASTM Committee E34 on
during which chemical energy is converted to light energy.
Occupational Health and Safety and is the direct responsibility of Subcommittee
E34.50 on Health and Safety Standards for Metal Working Fluids.
3.6 biomass, n—any matter which is or was a living
Current edition approved Aug. 1, 2011. Published August 2011. Originally
organism or excreted from a microorganism (D6161).
approved in 2009. Last previous edition approved in 2009 as E2694 - 09.
DOI:10.1520/E2694-11.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
3
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM AvailablefromU.S.GovernmentPrintingOfficeSuperintendentofDocuments,
Standards volume information, refer to the standard’s Document Summary page on 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
the ASTM website. 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 − 11
3.7 culturable, adj—microorganisms that proliferate as in- indicator of total microbial contamination in metalworking
dicated by the formation of colonies on solid growth media or fluids. ATP is not associated with m
...
This document is not anASTM standard and is intended only to provide the user of anASTM 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.
An American National Standard
Designation:E2694–09 Designation:E2694–11
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 The method provides a protocol for capturing, extracting and quantifying the adenosine triphosphate (ATP) content
associated with microorganisms found in water-miscible metalworking fluids (MWF).
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) which are
converted by comparison with an ATP standard and computation to pg ATP/mL.
1.3 This method is equally suitable for use in the laboratory or field.
1.4 The 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 betweenATPfrom different sources, for example, from different types of microorganisms,
such as bacteria and fungi.
1.6 The values stated in SI are to be regarded as 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
2
2.1 ASTM Standards:
D1129 Terminology Relating to Water
D4012 Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in Water
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 Nonconventional Microbiological Tests Used for Enumerating Bacteria
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 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.
3.2 adenosine triphosphate (ATP), n—a molecule comprised of a purine and three phosphate groups that serves as the primary
energy transport molecule in all biological cells.
1
This test method is under the jurisdiction ofASTM 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 May 1, 2009. Published June 2009. DOI: 10.1520/E2694-09.
Current edition approvedAug. 1, 2011. PublishedAugust 2011. Originally approved in 2009. Last previous edition approved in 2009 as E2694 - 09. DOI:10.1520/E2694-
11.
2
For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM 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.
3
Available from U.S. Government Printing Office Superintendent of Documents, 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
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 ----------------------
E2694–11
3.3 adenosine monophosphate (AMP), n—the molecule formed by the removal of two molecules of phosphate (one
pyrophosphate molecule) from ATP.
3.4 aseptic, adj—sterile, free from viable microbial contamination.
3.5 bioluminescence, n—the production and emission of light by a living organism as the result of a chemical reaction during
whic
...
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