ASTM D4200-82(1998)

NOTICE: This standard has either been superceded and replaced by a new version or discontinued.
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Designation: D 4200 – 82 (Reapproved 1998)
Standard Test Method for
Evaluating Inhibitory Effects of Ink Grids on Membrane
Filters
This standard is issued under the fixed designation D 4200; 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 5.2 A particularly sensitive organism and growth conditions
have been selected for this test method in order to maximize
1.1 This test method describes a procedure whereby the user
sensitivity to toxic materials possibly present in the inks used
of ink-gridded membrane filters in water quality studies can
for grid-marking membrane filters.
ascertain whether or not the grid lines are toxic and inhibitory
to bacterial growth when the membrane and its entrapped
6. Apparatus
bacteria are incubated on a suitable media.
6.1 Incubator, capable of maintaining temperatures of
1.2 This standard does not purport to address all of the
44.56 0.2°C.
safety concerns, if any, associated with its use. It is the
6.2 Membrane Filtration Units.
responsibility of the user of this standard to establish appro-
6.3 Vacuum Source with trap vessel.
priate safety and health practices and determine the applica-
6.4 Forceps, blunt-nosed.
bility of regulatory limitations prior to use.
6.5 Autoclave or other sterilizing equipment.
2. Referenced Documents 6.6 Expendables:
6.6.1 Gridded membrane filters.
2.1 ASTM Standards:
6.6.2 1-mL and 10-mL pipets.
D 1129 Terminology Relating to Water
6.6.3 Petri dishes (50-mm) containing 6 to 8 mL of agar
D 1193 Specification for Reagent Water
medium or a 100-mm dish with 20 6 2 mL of agar medium, or
3. Terminology
both.
6.6.4 Erlenmeyer flasks.
3.1 Definitions—For definitions of terms used in this test
method, refer to Terminology D 1129.
7. Reagents and Materials
4. Summary of Test Method
7.1 Purity of Water—Unless otherwise indicated, reference
to water shall be understood to mean reagent water conforming
4.1 A heavy bacterial suspension is filtered through a
to reagent water Type II of Specification D 1193.
gridded membrane filter. The bacterial concentration employed
7.2 M-FC Agar with Rosolic Acid or equivalent (henceforth
is sufficient to cover much of the membrane with bacterial
referred to as agar medium), formulated, prepared, and dis-
colonies.
pensed in accordance with the manufacturer’s specifications.
4.2 After filtration the membrane is incubated on a suitable
7.3 Tryptone Soya Broth or equivalent (henceforth referred
medium and the distribution of the colonies and shape of the
to as broth medium), formulated, prepared, and dispensed in
colonies noted in the area around each grid line.
accordance with the manufacturer’s specifications.
5. Significance and Use
7.4 Peptone Water, 0.1 %, sterile.
7.5 Broth Culture of E. coli ATCC 11229, 18-h, prepared as
5.1 This test method may be applied to determine the
follows: Add 1 mL of an 18 6 2-h broth culture of E. coli
suitability of grid-marked membrane filters for use in bacte-
ATCC 11229 to 99 mL of 0.1 % peptone water, mix thor-
riological culture techniques for the detection and enumeration
oughly, then add 0.1 mL of this suspension to another flask
of bacter
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