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ASTM D3731-87(2012)

(Practice)

Standard Practices for Measurement of Chlorophyll Content of Algae in Surface Waters

Standard Practices for Measurement of Chlorophyll Content of Algae in Surface Waters

SIGNIFICANCE AND USE
4.1 Data on the chlorophyll content of the algae have the following applications:  
4.1.1 To provide estimates of algal biomass and productivity.  
4.1.2 To provide general information on the taxonomic composition (major groups) of the algae, based on the relative amounts of chlorophyll a, b, and c, and the physiological condition of algal communities, which is related to the relative abundance of pheopigments.  
4.1.3 To determine long-term trends in water quality.  
4.1.4 To determine the trophic status of surface waters.  
4.1.5 To detect adverse effects of pollutants on plankton and periphyton in receiving waters.  
4.1.6 To determine maximum growth rates and yields in algal growth potential tests.
SCOPE
1.1 These practices include the extraction and the measurement of chlorophyll a, b, and  c, and pheophytin a in freshwater and marine plankton and periphyton. Three practices are provided as follows:  
1.1.1 Spectrophotometric, trichromatic practice for measuring chlorophyll a, b, and c.  
1.1.2 Spectrophotometric, monochromatic practice for measuring chlorophyll a corrected for pheophytin a; and for measuring pheophytin a.  
1.1.3 Fluorometric practice for measuring chlorophyll a corrected for pheophytin a; and for measuring pheophytin a.  
1.2 The values stated in SI units are to be regarded as the standard. The values given in parentheses are provided for information purposes only.  
1.3 This standard does not purport to address all of the safety problems, 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. Specific precautionary statements are given in Section 7.

General Information

Status
Historical
Publication Date
31-Aug-2012
ICS
13.060.10 - Water of natural resources
Technical Committee
D19 - Water
Drafting Committee
D19.24 - Water Microbiology
Current Stage
Ref Project

Relations

Replaced By
ASTM D3731-20 - Standard Practices for Measurement of Chlorophyll Content of Algae in Surface Waters
Effective Date
01-Jun-2020
Referred By
ASTM E645-18 - Standard Practice for Evaluation of Microbicides Used in Cooling Water Systems
Effective Date
01-Sep-2012
Referred By
ASTM E1218-21 - Standard Guide for Conducting Static Toxicity Tests with Microalgae
Effective Date
01-Sep-2012

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ASTM D3731-87(2012) - Standard Practices for Measurement of Chlorophyll Content of Algae in Surface WatersASTM D3731-87(2012) - Standard Practices for Measurement of Chlorophyll Content of Algae in Surface Waters
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ASTM D3731-87(2012) - Standard Practices for Measurement of Chlorophyll Content of Algae in Surface Waters
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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: D3731 − 87 (Reapproved 2012)
Standard Practices for Measurement of
1
Chlorophyll Content of Algae in Surface Waters
This standard is issued under the fixed designation D3731; 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 3. Summary of Practices
1.1 These practices include the extraction and the measure- 3.1 The chlorophyll and related compounds are extracted
mentofchlorophyll a, b,and c,andpheophytin ainfreshwater from the algae with 90% aqueous acetone. The concentration
and marine plankton and periphyton. Three practices are of the pigments is determined by measuring the light absorp-
provided as follows: tion or fluorescence of the extract.
1.1.1 Spectrophotometric, trichromatic practice for measur-
4. Significance and Use
ing chlorophyll a, b, and c.
1.1.2 Spectrophotometric,monochromaticpracticeformea-
4.1 Data on the chlorophyll content of the algae have the
suring chlorophyll a corrected for pheophytin a; and for
following applications:
measuring pheophytin a.
4.1.1 To provide estimates of algal biomass and productiv-
1.1.3 Fluorometric practice for measuring chlorophyll a
ity.
corrected for pheophytin a; and for measuring pheophytin a.
4.1.2 To provide general information on the taxonomic
composition (major groups) of the algae, based on the relative
1.2 The values stated in SI units are to be regarded as the
amounts of chlorophyll a, b, and c, and the physiological
standard. The values given in parentheses are provided for
condition of algal communities, which is related to the relative
information purposes only.
abundance of pheopigments.
1.3 This standard does not purport to address all of the
4.1.3 To determine long-term trends in water quality.
safety concerns, if any, associated with its use. It is the
4.1.4 To determine the trophic status of surface waters.
responsibility of the user of this standard to establish appro-
4.1.5 Todetectadverseeffectsofpollutantsonplanktonand
priate safety, health, and environmental practices and deter-
periphyton in receiving waters.
mine the applicability of regulatory limitations prior to use.
4.1.6 To determine maximum growth rates and yields in
Specific precautionary statements are given in Section 7.
algal growth potential tests.
1.4 This international standard was developed in accor-
dance with internationally recognized principles on standard-
5. Interferences and Special Considerations
ization established in the Decision on Principles for the
5.1 Pigment Extraction—The chlorophylls are only poorly
Development of International Standards, Guides and Recom-
extracted, if at all, from some forms of algae, such as the
mendations issued by the World Trade Organization Technical
coccoid green algae, unless the cells are disrupted, whereas
Barriers to Trade (TBT) Committee.
other algae, such as the diatoms, give up their pigments very
readily when merely steeped in acetone. Since natural commu-
2. Terminology
nities of algae usually consist of a wide variety of taxa that
2.1 Definitions:
differ in their resistance to extraction, it is necessary to disrupt
2.1.1 plankton—nonmotileorweaklyswimmingorganisms,
the cells routinely to ensure maximum recovery of the chloro-
usually microscopic, that drift or are carried along by currents
phylls. Failure to do so may result in a systematic underesti-
in surface waters, commonly consisting of bacteria, algae,
mation of 10% or more in the chlorophyll content of the
protozoa, rotifers, and microcrustacea.
2
samples. (1, 2, 3)
2.1.2 periphyton—microorganisms growing on submerged
5.2 Grinders—The cells of many common coccoid green
objects, commonly consisting of bacteria, algae, protozoa, and
algae resist disruption by most methods, but usually yield their
rotifers.
pigments after maceration with a tissue grinder. The routine
use of grinders, therefore, is recommended. Glass-to-glass
grinders are more rigorous in disrupting cells in plankton
1
These practices are under the jurisdiction ofASTM Committee D19 on Water
and are the direct responsibility of Subcommittee D19.24 on Water Microbiology.
Current edition approved Sept. 1, 2012. Published October 2012. Originally
2
approved in 1979. Last previous edition approved in 2004 as D3731–87 (2004). Theboldfacenumbersinparenthesesrefertothelistofreferencesattheendof
DOI: 10.1520/D3731-87R12. this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
D3731 − 87 (2012)
concentrated by centrifugation, and in periphyton scraping
...

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