ASTM D3862-80(2001)
(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of 0.2-µm Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality (Withdrawn 2010)
Standard Test Method for Retention Characteristics of 0.2-µm Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality (Withdrawn 2010)
SIGNIFICANCE AND USE
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.
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 This standard does not purport to address the safety problems 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.
WITHDRAWN RATIONALE
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.
Formerly under the jurisdiction of Committee D19 on Water, this test method was withdrawn in July 2010 in accordance with section 10.5.3.1 of the Regulations Governing ASTM Technical Committees, which requires that standards shall be updated by the end of the eighth year since the last approval date.
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Designation:D3862–80 (Reapproved 2001)
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 3.2.2 vacuum—for the procedure used—source of suction
that can produce a reading of 500 to 600 mm Hg on a vacuum
1.1 This test method covers a procedure to test membrane
gage.
filtersfortheirabilitytoretainbacteriawhosediameterisequal
to or slightly larger than the 0.2-µm pore size of the membrane
4. Summary of Test Method
filter.
4.1 This test method is based on the cultivation of organ-
1.2 The procedures described are for the use of user
isms whose diameters are equal to or slightly larger than pores
laboratories as differentiated from manufacturers’ laboratories.
of the membrane filter to be tested and then filtering a specific
1.3 This standard does not purport to address all of the
aliquot containing organisms through the membrane followed
safety concerns, if any, associated with its use. It is the
by an examination of the filtrate after incubation for sterility.A
responsibility of the user of this standard to establish appro-
sterile filtrate indicates complete retention of the organism and
priate safety and health practices and determine the applica-
validates the ability of the membrane to retain bacteria equal to
bility of regulatory limitations prior to use.
or slightly larger than the stated pore size.
2. Referenced Documents
5. Significance and Use
2.1 ASTM Standards:
5.1 Microbiological water testing procedures using mem-
D1129 Terminology Relating to Water
brane filtration are based on the premise that all bacteria within
D1193 Specification for Reagent Water
a specific size range will be retained by the membrane filter
3. Terminology used.Ifthemembranefilterdoesnotretainthesebacteria,false
negative results or lowered density estimates may occur that
3.1 Definitions:
could have serious repercussions due to the presence of
3.1.1 For definitions of terms used in this test method, refer
unrecognizedpotentialhealthhazardsinthewaterbeingtested,
to Terminology D1129.
especially in drinking water.
3.2 Definitions of Terms Specific to This Standard:
5.1.1 This procedure as devised will enable the user to test
3.2.1 Gram’s stain—a routine bacterial stain that divides
each membrane filter lot number for its ability to retain all
bacteria into two categories, depending on whether they can be
bacterial equal to, or larger than, the stated membrane pore
decolorized with acetone, alcohol, or aniline oil after staining
size.
with one of the rosaniline dyes such as crystal violet, methyl
5.2 Since this membrane is often used to sterilize nonauto-
violet, or gentian violet and treating with iodine. Those that
clavable liquids, it is essential that the retention characteristics
resist decolorization remain blue or violet and are designated
of this membrane are stable.
Gram-positive; those that are decolorized and take up the red
counterstain, such as neutral red, safranin, or dilute carbol
6. Apparatus
fuchsin are termed Gram-negative.
6.1 Membrane Filtration Units, six.
6.2 Vacuum Source, with trap vessel.
This test method is under the jurisdiction of ASTM Committee D19 on Water
6.3 Filtering flasks, 1-L, with vacuum tubing into which a
and is the direct responsibility of Subcommittee D19.08 on Membranes and Ion
glasstubeandaY-tubehavebeenincorporatedasinFig.1.The
Exchange Materials.
free end of the Y-tube is connected by tubing to a sterile
Current edition approved Feb. 12, 1980. Published April 1980. DOI: 10.1520/
D3862-80R01.
bacterial air vent.The tubing to air vent is clamped shut during
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
filtration and released after filtration.
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
6.4 Forceps,blunt-nosed,andsmallbeakerof95 %ethanol.
Standards volume information, refer to the standard’s Document Summary page on
6.5 Incubator, 37°C.
the ASTM website.
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D3862–80 (2001)
FIG. 1 Apparatus Required for Testing Retention Characteristics of Membrane Filters
6.6 Pinch-Cock Clamps. 24 62hsalinelactosebrothcultureto99mLof0.1 %peptone
6 7
6.7 Autoclave or Other Sterilizing Equipment. water. This suspension will contain approximately 10 to 10
6.8 Appropriate Equipment for producing reagent grade organisms per millilitre.
waters.
7.4 Peptone Water (0.1 %)—Prepare a 10 % stock solution
6.9 Appropriate Laboratory Glassware. of peptone in water. Dilute a measured volume of the 10 %
6.10 Sterile Rubber Stoppers, to fit 1-L filtering flask.
stock solution to obtain final solution of 0.1 % peptone in
6.11 Expendables: required amount. Sterilize at 121°C for 15 min.
6.11.1 Double-Strength Broth, 140-mL aliquots.
7.5 Test Organism—Pseudomonas diminuta ATCC strain
6.11.2 Sterile Pipets, 1 and 10-mL.
19146, also called FDA strain PC1—818.
6.11.3 Sterile 0.1 % Peptone, in 99-mL quantities.
7.6 Tryptic Soy Agar and Tryptone Soya Agar are inter-
6.11.4 Sterile 0.1 % Peptone, as rinse water.
changeable and henceforth referred to as agar medium, formu-
6.11.5 Broth Culture of Pseudomonas diminuta,24 62h.
lated, prepared, and dispensed in accordance with the manu-
6.11.6 Sterile Membrane Filters—Test membranes.
facturer’s specifications.
6.11.7 Petri Dishes, 50-mm, containing 6 to 8 mL of agar.
7.7 Tryptone Soya and Tryptic Soy Broth are interchange-
able and henceforth referred to as broth medium, formulated,
7. Reagents and Materials
prepared, and dispensed in accordance with the manufacturer’s
7.1 Purity of Reagents—Reagent grade chemicals shall be
specifications.
used in all tests. Unless otherwise indicated, it is intended that
all reagents shall conform to the specifications of the Commit-
8. Procedure
tee onAnalytical Reagents of the
...
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