ASTM D3862-13(2019)
(Test Method)Standard Test Method for Retention Characteristics of 0.2-µm Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality
Standard Test Method for Retention Characteristics of 0.2-µm Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality
SIGNIFICANCE AND USE
5.1 Microbiological water testing procedures using membrane filtration are based on the premise that all bacteria within a specific size range will be retained by the membrane filter used. If the membrane filter does not retain these bacteria, false negative results or lowered density estimates may occur that could have serious repercussions due to the presence of unrecognized potential health hazards in the water being tested, especially in drinking water.
5.1.1 This procedure as devised will enable the user to test each membrane filter lot number for its ability to retain all bacterial equal to, or larger than, the stated membrane pore size.
5.2 Since this membrane is often used to sterilize nonautoclavable liquids, it is essential that the retention characteristics of this membrane are stable.
SCOPE
1.1 This test method covers a procedure to test membrane filters for their ability to retain bacteria whose diameter is equal to or slightly larger than the 0.2-µm pore size of the membrane filter.
1.2 The procedures described are for the use of user laboratories as differentiated from manufacturers’ laboratories.
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.4 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.
1.5 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
Relations
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: D3862 − 13 (Reapproved 2019)
Standard Test Method for
Retention Characteristics of 0.2-μm Membrane Filters Used
in Routine Filtration Procedures for the Evaluation of
Microbiological Water Quality
This standard is issued under the fixed designation D3862; 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 3.2 Definitions of Terms Specific to This Standard:
3.2.1 Gram’s stain, n—a routine bacterial stain that divides
1.1 This test method covers a procedure to test membrane
bacteria into two categories, depending on whether they can be
filters for their ability to retain bacteria whose diameter is equal
decolorized with acetone, alcohol, or aniline oil after staining
to or slightly larger than the 0.2-μm pore size of the membrane
with one of the rosaniline dyes such as crystal violet, methyl
filter.
violet, or gentian violet and treating with iodine. Those that
1.2 The procedures described are for the use of user
resist decolorization remain blue or violet and are designated
laboratories as differentiated from manufacturers’ laboratories.
Gram-positive; those that are decolorized and take up the red
1.3 The values stated in SI units are to be regarded as counterstain, such as neutral red, safranin, or dilute carbol
standard. No other units of measurement are included in this fuchsin are termed Gram-negative.
standard.
3.2.2 vacuum, n—for the procedure used, source of suction
1.4 This standard does not purport to address all of the that can produce a reading of 500 to 600 mm Hg on a vacuum
safety concerns, if any, associated with its use. It is the gage.
responsibility of the user of this standard to establish appro-
priate safety, health, and environmental practices and deter-
4. Summary of Test Method
mine the applicability of regulatory limitations prior to use.
4.1 This test method is based on the cultivation of organ-
1.5 This international standard was developed in accor-
isms whose diameters are equal to or slightly larger than pores
dance with internationally recognized principles on standard-
of the membrane filter to be tested and then filtering a specific
ization established in the Decision on Principles for the
aliquot containing organisms through the membrane followed
Development of International Standards, Guides and Recom-
by an examination of the filtrate after incubation for sterility. A
mendations issued by the World Trade Organization Technical
sterile filtrate indicates complete retention of the organism and
Barriers to Trade (TBT) Committee.
validates the ability of the membrane to retain bacteria equal to
or slightly larger than the stated pore size.
2. Referenced Documents
2.1 ASTM Standards:
5. Significance and Use
D1129 Terminology Relating to Water
5.1 Microbiological water testing procedures using mem-
D1193 Specification for Reagent Water
brane filtration are based on the premise that all bacteria within
a specific size range will be retained by the membrane filter
3. Terminology
used. If the membrane filter does not retain these bacteria, false
3.1 Definitions:
negative results or lowered density estimates may occur that
3.1.1 For definitions of terms used in this standard, refer to
could have serious repercussions due to the presence of
Terminology D1129.
unrecognized potential health hazards in the water being tested,
especially in drinking water.
5.1.1 This procedure as devised will enable the user to test
This test method is under the jurisdiction of ASTM Committee D19 on Water
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
each membrane filter lot number for its ability to retain all
Current edition approved Nov. 1, 2019. Published December 2019. Originally
bacterial equal to, or larger than, the stated membrane pore
approved in 1990. Last previous edition approved in 2013 as D3862 – 13. DOI:
size.
10.1520/D3862-13R19.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
5.2 Since this membrane is often used to sterilize nonauto-
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
clavable liquids, it is essential that the retention characteristics
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. of this membrane are stable.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D3862 − 13 (2019)
6. Apparatus where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of
6.1 Membrane Filtration Units, six.
sufficiently high purity to permit its use without lessening the
6.2 Vacuum Source, with trap vessel. accuracy of the determination.
7.2 Purity of Water—Unless otherwise indicated, reference
6.3 Filtering flasks, 1-L, with vacuum tubing into which a
to water shall be understood to mean reagent water conforming
glass tube and a Y-tube have been incorporated as in Fig. 1. The
to Specification D1193, Type II, for 0.2-μm membrane filtra-
free end of the Y-tube is connected by tubing to a sterile
tion. In addition, suitability tests for determining the bacteri-
bacterial air vent. The tubing to air vent is clamped shut during
cidal properties of the reagent grade water should be per-
filtration and released after filtration.
formed.
6.4 Forceps, blunt-nosed, and small beaker of 95 % ethanol.
7.3 Bacterial Suspension —Prepare 100 mL of a suspension
6.5 Incubator, 37°C.
of a Pseudomonas diminuta (ATCC 19146). Add 1.0 mL of a
24 6 2 h saline lactose broth culture to 99 mL of 0.1 % peptone
6.6 Pinch-Cock Clamps.
water. This suspension will contain approximately 106 to 107
6.7 Autoclave or Other Sterilizing Equipment.
organisms per millilitre.
6.8 Appropriate Equipment for producing reagent grade
7.4 Peptone Water (0.1 %)—Prepare a 10 % stock solution
waters. of peptone in water. Dilute a measured volume of the 10 %
stock solution to obtain final solution of 0.1 % peptone in
6.9 Appropriate Laboratory Glassware.
required amount. Sterilize at 121°C for 15 min.
6.10 Sterile Rubber Stoppers, to fit 1-L filtering flask.
7.5 Test Organism—Pseudomonas diminuta ATCC strain
6.11 Expendables: 19146, also called FDA strain PC1—818.
6.11.1 Double-Strength Broth, 140-mL aliquots.
7.6 Tryptic Soy Agar and Tryptone Soya Agar are inter-
6.11.2 Sterile Pipets, 1 and 10-mL.
changeable and henceforth referred to as
...
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