ASTM D3863-87(2003)
(Test Method)Standard Test Method for Retention Characteristics of 0.40 to 0.45-µm Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality
Standard Test Method for Retention Characteristics of 0.40 to 0.45-µm Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality
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.
This procedure as devised will enable the user to test each membrane filter lot number for its ability to retain all bacteria equal to, or larger than, the stated membrane pore size.
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 membrane filters with pore size rated at 0.40 to 0.45 μm.
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 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:D3863–87(Reapproved2003)
Standard Test Method for
Retention Characteristics of 0.40 to 0.45-µm Membrane
Filters Used in Routine Filtration Procedures for the
Evaluation of Microbiological Water Quality
This standard is issued under the fixed designation D3863; 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 counterstain, such as neutral red, safranin, or dilute carbol
fuchsin are termed Gram-negative.
1.1 This test method covers a procedure to test membrane
3.2.2 vacuum—for the procedure used, a source of suction
filtersfortheirabilitytoretainbacteriawhosediameterisequal
that can produce a reading of 500 to 600 mm Hg on a vacuum
to or slightly larger than membrane filters with pore size rated
gage.
at 0.40 to 0.45 µm.
1.2 The procedures described are for the use of user
4. Summary of Test Method
laboratories as differentiated from manufacturers’ laboratories.
4.1 This test method is based on the cultivation of organ-
1.3 This standard does not purport to address all of the
isms whose diameters are equal to or slightly larger than pores
safety concerns, if any, associated with its use. It is the
of the membrane filter to be tested and then filtering a specific
responsibility of the user of this standard to establish appro-
aliquot containing organisms through the membrane followed
priate safety and health practices and determine the applica-
by an examination of the filtrate after incubation for sterility.A
bility of regulatory limitations prior to use.
sterile filtrate indicates complete retention of the organism and
2. Referenced Documents validates the ability of the membrane to retain bacteria equal to
or slightly larger than the stated pore size.
2.1 ASTM Standards:
D1129 Terminology Relating to Water
5. Significance and Use
D1193 Specification for Reagent Water
5.1 Microbiological water testing procedures using mem-
3. Terminology brane filtration are based on the premise that all bacteria within
a specific size range will be retained by the membrane filter
3.1 Definitions:
used.Ifthemembranefilterdoesnotretainthesebacteria,false
3.1.1 For definitions of terms used in this method refer to
negative results or lowered density estimates may occur that
Terminology D1129.
could have serious repercussions due to the presence of
3.2 Definitions of Terms Specific to This Standard:
unrecognizedpotentialhealthhazardsinthewaterbeingtested,
3.2.1 Gram’s stain—a routine bacterial stain that divides
especially in drinking water.
bacteria into two categories, depending on whether they can be
5.2 This procedure as devised will enable the user to test
decolorized with acetone, alcohol, or aniline oil after staining
each membrane filter lot number for its ability to retain all
with one of the rosaniline dyes such as crystal violet, methyl
bacteriaequalto,orlargerthan,thestatedmembraneporesize.
violet, or gentian violet and treating with iodine. Those that
resist decolorization remain blue or violet and are designated
6. Apparatus
Gram-positive; those that are decolorized and take up the red
6.1 Membrane Filtration Units, six.
6.2 Vacuum Source with trap vessel.
6.3 Filtering Flasks, 1-L, with vacuum tubing into which a
This test method is under the jurisdiction of ASTM Committee D19 on Water
glasstubeandaY-tubehavebeenincorporatedasinFig.1.The
and is the direct responsibility of Subcommittee D19.08 on Membranes and Ion
free end of the Y-tube is connected by tubing to a sterile
Exchange Materials.
bacterial air vent.The tubing to air vent is clamped shut during
Current edition approved March 27, 1987. Published July 1987. DOI: 10.1520/
D3863-87R03.
filtration and released after filtration.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
6.4 Forceps,blunt-nosed,andsmallbeakerof95 %ethanol.
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
6.5 Incubator, 20 to 25°C.
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. 6.6 Pinch-Cock Clamps.
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D3863–87 (2003)
FIG. 1 Apparatus Required for Testing Retention Characteristics of Membrane Filters
6.7 Autoclave or Other Sterilizing Equipment. 7.4 Peptone Water (0.1 %)—Prepare a 10 % stock solution
6.8 Appropriate Equipment for producing reagent grade of peptone in water. Dilute a measured volume of the 10 %
water. 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—Serratia marcescens ATCC strain
6.11 Expendables:
14756, also called FDA strain PC1 1107. Red pigmented
6.11.1 Double-Strength Broth, 140-mL aliquots.
Serratia marcescens strains other than the above may also be
6.11.2 Sterile Pipets, 1 and 10-mL.
used.
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 Cultures of Serratia marcescens,18 62h.
lated, prepared, and dispensed in accordance with the manu-
6.11.6 Sterile Membrane Filters—Test membranes.
facturers’ 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 manufacturers’
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 theAmerican
...
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