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ASTM D8243-19

(Test Method)

Standard Test Method for Determination of APS Reductase to Estimate Sulfate Reducing Bacterial Bioburdens in Water – Enzyme-Linked Immunosorbent Assay Method

Standard Test Method for Determination of APS Reductase to Estimate Sulfate Reducing Bacterial Bioburdens in Water – Enzyme-Linked Immunosorbent Assay Method

SIGNIFICANCE AND USE
5.1 Sulfate reducing archaea and bacteria are known to contribute to microbiologically influenced corrosion.  
5.2 Sulfate-reducing bacteria are widely distributed in marine and fresh water muds which, in consequence, frequently are laden with the hydrogen sulfide produced by these organisms during dissimilatory sulfate reduction.  
5.3 Traditional, culture-dependent methods such as those described in Test Methods D4412, prescribe incubation periods of as long as 21 days before assigning a below detection limit (BDL) score to a specimen. Moreover, it is well known that not all SRP will proliferate in the nutrient media specified in Test Methods D4412.  
5.4 This test method uses ELISA technology to provide semi-quantitative, culture-independent, SRP bioburden test results in less than 30 min.  
5.4.1 Because all the reagents and supplies used are non-hazardous and prepackaged for single test use, this test method does not require any apparatus other than a laboratory timer. Consequently, it can be performed at or near the point of sample collection.  
5.4.2 The opportunity to minimize the delay between sample collection, testing, and results availability translates into timely use of the data to drive preventive and corrective SRB control measures.
SCOPE
1.1 This test method provides a protocol for using enzyme-linked immunosorbent assay (ELISA) technology to test water samples for the enzyme adenosine 5’-phosphosulfate reductase (APSr) concentration.  
1.1.1 APSr is present in all known sulfate reducing protists (SRP – sulfate reducing bacteria – SRB – and sulfate reducing archaea – SRA).  
1.1.2 As reported in U.S. Patent 4,999,286, APS reductase concentration can be used as a surrogate parameter for estimating SRA bioburdens (Appendix X1 compares results from Test Methods D8243, D4412, and quantitative polymerase chain reaction – qPCR – testing).  
1.2 This test method has been validated in tap water, oilfield produced water (salinities ranging from 100 g L-1 to 600 g L-1), and fuel-associated water (commonly referred to as bottoms-water).  
1.3 This test method detects APS reductase semi-quantitatively in the range of 0.001M to 0.1M – correlating to 102 SRP/mL to 106 SRP/mL.  
1.3.1 As described in Appendix X2 test method sensitivity can be increased 10-fold to 100-fold. However, the precision statistics provided in X apply only to 10-mL specimens.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 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. Some specific hazards statements are given in Section 9 on Hazards.  
1.6 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

Status
Published
Publication Date
31-Dec-2018
ICS
07.100.20 - Microbiology of water
Technical Committee
D19 - Water
Drafting Committee
D19.24 - Water Microbiology
Current Stage
Ref Project

Relations

Refers
ASTM D6300-15 - Standard Practice for Determination of Precision and Bias Data for Use in Test Methods for Petroleum Products and Lubricants
Effective Date
01-Jun-2015
Refers
ASTM D6300-16 - Standard Practice for Determination of Precision and Bias Data for Use in Test Methods for Petroleum Products and Lubricants
Effective Date
01-Apr-2016
Refers
ASTM D4412-19 - Standard Test Methods for Sulfate-Reducing Bacteria in Water and Water-Formed Deposits
Effective Date
01-Dec-2019
Refers
ASTM E1601-19 - Standard Practice for Conducting an Interlaboratory Study to Evaluate the Performance of an Analytical Method
Effective Date
01-Nov-2019
Refers
ASTM D6300-19a - Standard Practice for Determination of Precision and Bias Data for Use in Test Methods for Petroleum Products and Lubricants
Effective Date
01-Dec-2019
Refers
ASTM D7464-19 - Standard Practice for Manual Sampling of Liquid Fuels, Associated Materials and Fuel System Components for Microbiological Testing
Effective Date
01-Nov-2019
Refers
ASTM D7464-20 - Standard Practice for Manual Sampling of Liquid Fuels, Associated Materials and Fuel System Components for Microbiological Testing
Effective Date
01-May-2020
Refers
ASTM D7464-14 - Standard Practice for Manual Sampling of Liquid Fuels, Associated Materials and Fuel System Components for Microbiological Testing
Effective Date
01-Oct-2014

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ASTM D8243-19 - Standard Test Method for Determination of APS Reductase to Estimate Sulfate Reducing Bacterial Bioburdens in Water – Enzyme-Linked Immunosorbent Assay MethodASTM D8243-19 - Standard Test Method for Determination of APS Reductase to Estimate Sulfate Reducing Bacterial Bioburdens in Water – Enzyme-Linked Immunosorbent Assay Method
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ASTM D8243-19 - Standard Test Method for Determination of APS Reductase to Estimate Sulfate Reducing Bacterial Bioburdens in Water – Enzyme-Linked Immunosorbent Assay Method
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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: D8243 − 19
Standard Test Method for
Determination of APS Reductase to Estimate Sulfate
Reducing Bacterial Bioburdens in Water – Enzyme-Linked
1
Immunosorbent Assay Method
This standard is issued under the fixed designation D8243; 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 ization established in the Decision on Principles for the
Development of International Standards, Guides and Recom-
1.1 This test method provides a protocol for using enzyme-
mendations issued by the World Trade Organization Technical
linked immunosorbent assay (ELISA) technology to test water
Barriers to Trade (TBT) Committee.
samplesfortheenzymeadenosine5’-phosphosulfatereductase
(APSr) concentration.
2. Referenced Documents
1.1.1 APSr is present in all known sulfate reducing protists
2
2.1 ASTM Standards:
(SRP– sulfate reducing bacteria – SRB – and sulfate reducing
D1129Terminology Relating to Water
archaea – SRA).
D4412TestMethodsforSulfate-ReducingBacteriainWater
1.1.2 As reported in U.S. Patent 4,999,286, APS reductase
and Water-Formed Deposits
concentration can be used as a surrogate parameter for esti-
D5847Practice for Writing Quality Control Specifications
mating SRAbioburdens (Appendix X1 compares results from
for Standard Test Methods for Water Analysis
Test Methods D8243, D4412, and quantitative polymerase
D6300Practice for Determination of Precision and Bias
chain reaction – qPCR – testing).
Data for Use in Test Methods for Petroleum Products and
1.2 Thistestmethodhasbeenvalidatedintapwater,oilfield
Lubricants
-1 -1
producedwater(salinitiesrangingfrom100gL to600gL ),
D6499Test Method for Immunological Measurement of
and fuel-associated water (commonly referred to as bottoms-
AntigenicProteininHeveaNaturalRubber(HNR)andits
water).
Products
1.3 This test method detects APS reductase semi-
D7464Practice for Manual Sampling of Liquid Fuels, As-
quantitatively in the range of 0.001M to 0.1M – correlating to
sociated Materials and Fuel System Components for
2 6
10 SRP/mL to 10 SRP/mL.
Microbiological Testing
1.3.1 As described in Appendix X2 test method sensitivity
E691Practice for Conducting an Interlaboratory Study to
can be increased 10-fold to 100-fold. However, the precision
Determine the Precision of a Test Method
statistics provided in X apply only to 10-mL specimens.
E1601Practice for Conducting an Interlaboratory Study to
Evaluate the Performance of an Analytical Method
1.4 The values stated in SI units are to be regarded as
E1847Practice for Statistical Analysis of Toxicity Tests
standard. No other units of measurement are included in this
Conducted Under ASTM Guidelines
standard.
E1914Practice for Use of Terms Relating to the Develop-
1.5 This standard does not purport to address all of the
ment and Evaluation of Methods for Chemical Analysis
safety concerns, if any, associated with its use. It is the
3
(Withdrawn 2016)
responsibility of the user of this standard to establish appro-
4
2.2 Patents:
priate safety, health, and environmental practices and deter-
U.S. Patent 4,999,286Sulfate Reducing Bacteria Determi-
mine the applicability of regulatory limitations prior to use.
nation and Control, March 12, 1991
Some specific hazards statements are given in Section 9 on
Hazards.
1.6 This international standard was developed in accor-
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
dance with internationally recognized principles on standard-
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.
1 3
This test method is under the jurisdiction ofASTM Committee D19 on Water The last approved version of this historical standard is referenced on
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology. www.astm.org.
4
Current edition approved Jan. 1, 2019. Published February 2019. DOI: 10.1520/ Available from United States Patent and Trademark Office (USPTO), Madison
D8243-19. Building, 600 Dulany Street, Alexandria, VA 22314, https://www.uspto.gov.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
D8243 − 19
3. Terminology 4.7 At the end of the color-development period (that is, 15
min at 18 6 2°C), compare and match the color of the funnel
3.1 Definitions:
5
d
...

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1.4 The values stated in SI units are to be regarded as the standard. No other units of measurement are included in this standard.  
1.5 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.  
1.6 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.

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