ASTM D3863-87(1998)
(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
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 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.
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Standards Content (Sample)
NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
Designation: D 3863 – 87 (Reapproved 1998)
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 D 3863; 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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope 4. Summary of Test Method
1.1 This test method covers a procedure to test membrane 4.1 This test method is based on the cultivation of organ-
filters for their ability to retain bacteria whose diameter is equal isms whose diameters are equal to or slightly larger than pores
to or slightly larger than membrane filters with pore size rated of the membrane filter to be tested and then filtering a specific
at 0.40 to 0.45 μm. aliquot containing organisms through the membrane followed
1.2 The procedures described are for the use of user by an examination of the filtrate after incubation for sterility. A
laboratories as differentiated from manufacturers’ laboratories. sterile filtrate indicates complete retention of the organism and
1.3 This standard does not purport to address all of the validates the ability of the membrane to retain bacteria equal to
safety problems, if any, associated with its use. It is the or slightly larger than the stated pore size.
responsibility of the user of this standard to establish appro-
5. Significance and Use
priate safety and health practices and determine the applica-
bility of regulatory limitations prior to use. 5.1 Microbiological water testing procedures using mem-
brane filtration are based on the premise that all bacteria within
2. Referenced Documents
a specific size range will be retained by the membrane filter
2.1 ASTM Standards: used. If the membrane filter does not retain these bacteria, false
D 1129 Terminology Relating to Water negative results or lowered density estimates may occur that
D 1193 Specification for Reagent Water could have serious repercussions due to the presence of
unrecognized potential health hazards in the water being tested,
3. Terminology
especially in drinking water.
3.1 Definitions: 5.2 This procedure as devised will enable the user to test
3.1.1 For definitions of terms used in this method refer to
each membrane filter lot number for its ability to retain all
Terminology D 1129. bacteria equal to, or larger than, the stated membrane pore size.
3.2 Definitions of Terms Specific to This Standard:
6. Apparatus
3.2.1 Gram’s stain—a routine bacterial stain that divides
bacteria into two categories, depending on whether they can be 6.1 Membrane Filtration Units, six.
decolorized with acetone, alcohol, or aniline oil after staining 6.2 Vacuum Source with trap vessel.
with one of the rosaniline dyes such as crystal violet, methyl 6.3 Filtering Flasks, 1-L, with vacuum tubing into which a
violet, or gentian violet and treating with iodine. Those that glass tube and a Y-tube have been incorporated as in Fig. 1. The
resist decolorization remain blue or violet and are designated free end of the Y-tube is connected by tubing to a sterile
Gram-positive; those that are decolorized and take up the red bacterial air vent. The tubing to air vent is clamped shut during
counterstain, such as neutral red, safranin, or dilute carbol filtration and released after filtration.
fuchsin are termed Gram-negative. 6.4 Forceps, blunt-nosed, and small beaker of 95 % ethanol.
3.2.2 vacuum—for the procedure used, a source of suction 6.5 Incubator, 20 to 25°C.
that can produce a reading of 500 to 600 mm Hg on a vacuum 6.6 Pinch-Cock Clamps.
gage. 6.7 Autoclave or Other Sterilizing Equipment.
6.8 Appropriate Equipment for producing reagent grade
water.
This test method is under the jurisdiction of ASTM Committee D-19 on Water
6.9 Appropriate Laboratory Glassware.
and is the direct responsibility of Subcommittee D19.08 on Membranes and Ion
6.10 Sterile Rubber Stoppers, to fit 1-L filtering flask.
Exchange Materials.
6.11 Expendables:
Current edition approved March 27, 1987. Published July 1987.
1983 Annual Book of ASTM Standards, Vol 11.01. 6.11.1 Double-Strength Broth, 140-mL aliquots.
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NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
D 3863
FIG. 1 Apparatus Required for Testing Retention Characteristics of Membrane Filters
6.11.2 Sterile Pipets, 1 and 10-mL. 7.6 Tryptic Soy Agar and Tryptone Soya Agar—are inter-
6.11.3 Sterile 0.1 % Peptone in 99-mL quantities. changeable and henceforth referred to as agar medium, formu-
6.11.4 Sterile 0.1 % Peptone as rinse water. lated, prepared, and dispensed in accordance with the manu-
6.11.5 Broth Cultures of Serratia marcescens,18 6 2h. facturers’ specifications.
6.11.6 Sterile Membrane Filters—Test membranes.
7.7 Tryptone Soya and Tryptic Soy Broth—are interchange-
6.11.7 Petri Dishes, 50-mm, containing 6 to 8-mL of agar.
able and henceforth referred to as broth medium, formulated,
prepared, and dispensed in accordance with the manufacturers’
7. Reagents and Materials
specifications.
7.1 Purity of Reagents—Reagent grade chemicals shall be
used in all tests. Unless otherwise indicated, it is intended that
8. Procedure
all reagents shall conform to the specifications of the Commit-
8.1 Place 140 mL of double-strength broth into six 1-L
tee on Analytical Reagents of the American Chemical Society,
3 vacuum flasks with the attached vacuum tubing, Y-tube, and
where such specifications are available. Other grades may be
bacterial air vent. Wrap in kraft paper and sterilize by auto-
used, provided it is first ascertained that the reagent is of
claving at 121°C for 15 min.
sufficiently high purity to permit its use without lessening the
8.2 Aseptically assemble membrane filtration apparatus
accuracy of the determina
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