ASTM E2275-03

Standard Practice for Evaluating Water-Miscible Metalworking Fluid Bioresistance and Antimicrobial Pesticide Performance

SCOPE
1.1 This practice addresses the evaluation of the relative inherent bioresistance of water-miscible metalworking fluids, the bioresistance attributable to augmentation with antimicrobial pesticides or both. It replaces Methods D 3946 and E 686.
1.2 In this practice relative bioresistance is determined by challenging metalworking fluids with a biological inoculum that may either be characterized (comprised of one or more known biological cultures) or uncharacterized (comprised of biologically contaminated metalworking fluid or one or more unidentified isolates from deteriorated metalworking fluid). Challenged fluid bioresistance is defined in terms of resistance to biomass increase, viable cell recovery increase, chemical property change, physical property change or some combination thereof.
1.3 This practice is applicable to antimicrobial agents that are incorporated into either the metalworking fluid concentrate or end-use dilution. It is also applicable to metalworking fluids that are formulated using non-microbicidal, inherently bioresistant components.
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 and health practices and determine the applicability of regulatory limitations prior to use.

General Information

• Status: Historical
• Publication Date: 09-Apr-2003
• ICS: 75.100 - Lubricants, industrial oils and related products
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents
• Drafting Committee: E35.15 - Antimicrobial Agents

Relations

• Replaced By: ASTM E2275-03e1 - Standard Practice for Evaluating Water-Miscible Metalworking Fluid Bioresistance and Antimicrobial Pesticide Performance
• Effective Date: 10-Apr-2003
• Referred By: ASTM F86-21 - Standard Practice for Surface Preparation and Marking of Metallic Surgical Implants
• Effective Date: 10-Apr-2003
• Referred By: ASTM E2523-13(2018) - Standard Terminology for Metalworking Fluids and Operations
• Effective Date: 10-Apr-2003
• Referred By: ASTM D6666-20 - Standard Guide for Evaluation of Aqueous Polymer Quenchants
• Effective Date: 10-Apr-2003
• Referred By: ASTM E1497-23 - Standard Practice for Selection and Safe Use of Water-Miscible and Straight Oil Metal Removal Fluids
• Effective Date: 10-Apr-2003
• Referred By: ASTM E2889-12(2017) - Standard Practice for Control of Respiratory Hazards in the Metal Removal Fluid Environment
• Effective Date: 10-Apr-2003

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: E 2275 – 03
Standard Practice for
Evaluating Water-Miscible Metalworking Fluid Bioresistance
and Antimicrobial Pesticide Performance
This standard is issued under the fixed designation E 2275; 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 D 4012 Test Method for Adenosine Triphosphate (ATP)
Content of Microorganisms in Water
1.1 This practice addresses the evaluation of the relative
D 4627 Method for Iron Chip Corrosion for Water-
inherent bioresistance of water-miscible metalworking fluids,
Dilutable Metalworking Fluids
the bioresistance attributable to augmentation with antimicro-
D 5465 Practice for Determining Microbial Colony Counts
bial pesticides or both. It replaces Methods D 3946 and E 686.
from Waters Analyzed by Plating Methods
1.2 In this practice relative bioresistance is determined by
E 70 Test Method for pH of Aqueous Solutions with the
challenging metalworking fluids with a biological inoculum
Glass Electrode
that may either be characterized (comprised of one or more
E 1326 Guide for Evaluating Nonconventional Microbio-
known biological cultures) or uncharacterized (comprised of
logical tests Used for Enumerating Bacteria
biologically contaminated metalworking fluid or one or more
E 2169 Practice for Selecting Antimicrobial Pesticides for
unidentified isolates from deteriorated metalworking fluid).
use in Water-miscible Metalworking Fluids
Challenged fluid bioresistance is defined in terms of resistance
2.2 Other Standards:
to biomass increase, viable cell recovery increase, chemical
4.027 Synthetic Hard Water
property change, physical property change or some combina-
9215A.6a Heterotrophic Plate Count Media, Plate Count
tion thereof.
Agar
1.3 This practice is applicable to antimicrobial agents that
9216 Direct Total Microbial Count
are incorporated into either the metalworking fluid concentrate
Microbiological Test <71>
or end-use dilution. It is also applicable to metalworking fluids
2.3 Government Standard:
that are formulated using non-microbicidal, inherently biore-
40 CFR 156 Labeling Requirements for Pesticides and De-
sistant components.
vices
1.4 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
3. Terminology
responsibility of the user of this standard to establish appro-
3.1 Definitions:
priate safety and health practices and determine the applica-
3.1.1 active ingredient, n—the chemical component or com-
bility of regulatory limitations prior to use.
ponents of an antimicrobial pesticide that provides its micro-
2. Referenced Documents bicidal performance.
3.1.2 antimicrobial pesticide, n—chemical additive regis-
2.1 ASTM Standards:
2 tered under 40 CFR 152, for use to inhibit growth, proliferation
D 888 Test Methods for Dissolved Oxygen in Water
or both of microorganisms.
D 1067 Test Methods for Acidity or Alkalinity of Water
2 3.1.3 as supplied, adj—antimicrobial pesticide finished
D 1193 Specification for Reagent Grade Water
product including the active ingredient(s), solvent and any
D 3342 Method for Dispersion Stability of New (Unused)
3 additional inactive ingredients.
Rolling Oil Dispersions in Water
D 3519 Test Method for Foam in Aqueous Media (Blender
Test)
Annual Book of ASTM Standards, Vol 11.02.
D 3601 Test Method for Foam in Aqueous Media (Bottle
Annual Book of ASTM Standards, Vol 15.05.
Test)
Annual Book of ASTM Standards, Vol 11.05.
Annual Book of ASTM Standards, Vol 11.03.
AOAC International Methods of Analysis, AOAC International, Gaithersburg,
This practice is under the jurisdiction of ASTM Committee E35 on Geosyn- MD.
thetics and is the direct responsibility of Subcommittee E35.15 on Antimicrobial Available from American Public Health Association (APHA) Standard Meth-
Agents. ods for the Examination of Water and Wastewater 800 I Street, NW Washington, DC
Current edition approved April 10, 2003. Published May 2003. 20001.
2 10
Annual Book of ASTM Standards, Vol 11.01. Available from U.S. Pharmacopeia/National Formulary (USP/NF), 12601
Annual Book of ASTM Standards, Vol 05.02. Twinbrook Parkway Rockville, MD 20852.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
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.
E2275–03
3.1.4 biocide, n—any chemical intended for use to kill 4.3 Bioresistance is determined as the test fluid’s relative
organisms. ability to prevent the proliferation of challenge microbes, retain
3.1.5 bioresistant, adj—ability to withstand biological at- its original chemical or physical properties of some combina-
tack. tion of the above. The bioresistance of test formulations is
3.1.5.1 Discussion—Bioresistant, or recalcitrant, chemicals defined relative to that of a benchmark or control formulation.
are not readily metabolized by microorganisms.
5. Significance and Use
3.1.6 biostatic, adj—able to prevent existing microbial con-
5.1 This practice provides laboratory procedures for rating
taminants from growing or proliferating, but unable to kill
the relative bioresistance of metalworking fluid formulations,
them.
for determining the need for microbicide addition prior to or
3.1.6.1 Discussion—Biostatic additives may be registered
during fluid use in metalworking systems and for evaluating
antimicrobial pesticides or unregistered chemicals with other
microbicide performance. General considerations for microbi-
performance properties. The difference between biocidal and
cide selection are provided in Practice E 2169.
biostatic performance may be attributed to dose, chemistry or
5.2 The factors affecting challenge population numbers,
both.
taxonomic diversity, physiological state, inoculation frequency
3.1.7 dose, n—concentration of antimicrobial pesticide
and biodeterioration effects in recirculating metalworking fluid
added to treated solution.
systems are varied and only partially understood. Conse-
3.1.7.1 Discussion—Dose is generally expressed as either
quently, the results of tests completed in accordance with this
ppm active ingredient (a.i.) or ppm as supplied (a.s.).
practice should be used only to compare the relative perfor-
3.1.8 inactive ingredient, n—component of antimicrobial
mance of products or microbicide treatments included in a test
pesticide that is not directly responsible for the pesticide’s
antimicrobial performance. series. Results should not be construed as predicting actual
field performance.
3.1.8.1 Discussion—Inactive ingredients may include, but
are not limited to solvents and chemicals that improve the
6. Apparatus
pesticide’s non-biocidal performance properties, such as mis-
6.1 Air Supply, air provided at no more than 110 kPa.
cibility and reactivity with non-target molecules in the treated
material.
NOTE 1—Any air source that is free of organic vapors, organic matter
3.1.9 minimum inhibitory concentration (MIC), n—lowest
or other objectionable material may be used. Sterile air need not be used
treatment-dose that will prevent test population from growing, for the uncharacterized inoculum, but shall be used for the characterized
inoculum. If necessary, air may be sterilized either by inserting, in series,
proliferating or otherwise contributing to biodeterioration.
two commercially available in-line sterile filters designed for this purpose.
3.2 Abbreviations:
Alternatively an in-line filter may be prepared as follows: Pack two 150
3.2.1 a.i.—active ingredient
mm long drying tubes (bulb-type) loosely with borosilicate glass wool in
3.2.2 a.s.—as supplied
series with neoprene stoppers, glass tubing and neoprene tubing. Wrap
3.2.3 ATCC—american type culture collection
loosely in aluminum foil and steam sterilize at 103 to 138 kPa (15 to 20
3.2.4 CFU—colony forming unit
psi) for 30 min or dry heat sterilize at 160°C for 2 h. Cool to room
temperature while wrapped. Insert into air line with bulbs on upstream
4. Summary of Practice
side. Whether using a commercial or fabricated filter, average lifetime in
continuous use is two weeks. Discard sooner if upstream filter becomes
4.1 End-use dilutions of one or more water-miscible metal-
wet or contaminated with oil.
working fluids are dispensed into microcosms. The fluids may
be fresh or aged, dosed with one or more antimicrobial
6.2 Aquarium Tubing, 6.35 mm (0.25 in.) diameter, silicone
pesticides or undosed. Microcosms are challenged with either or vinyl.
uncharacterized or characterized biological inocula. After in-
6.3 Autoclave, with both steam cycle (80 to 100°C) and
oculation, microcosms are aerated either continuously or peri- sterilization cycle (15 min at $ 121°C) capability.
odically to simulate recirculation conditions in coolant sys-
6.4 Adjustable Volume Pipetters, with sterile disposable
tems. Chips may also be added to microcosms to simulate chip tips. Pipetters will be used to deliver 1.0 μL to 2 mL volumes.
accumulation in coolant systems.
6.5 Glassware:
4.2 After inoculation, fluid samples are drawn from each
NOTE 2—Sterile laboratory ware or sterile disposable laboratory ware
microcosm periodically and tested for the parameters of
should be used according to standard microbiological practice.
interest, including but not limited to microbial viable counts.
6.5.1 Glass Tubing, 6.35 mm (0.25 in.) i.d., cut into 15 cm
Depending on the test objectives, the test duration may range
lengths with ends fire-polished.
from 24 h to three months.
6.5.2 French Square Bottles, 960 mL, with metal cap.
4.2.1 Shorter test periods are used to evaluate microbicide
speed of kill and metalworking formulation initial bioresis-
NOTE 3—Alternatively, 1 L capacity canning jars may be used.
tance.
6.5.3 Pipetes, Bacteriological, 10 and 2.2 mL.
4.2.2 Longer test periods are used to evaluate metalworking
6.6 Incubator, capable of maintaining a temperature of 25 6
fluid formulation resistance to repeated challenges. For tests
2°C.
lasting longer than one-week, 10 to 80 % of the fluid is
NOTE 4—Although an incubator is preferred, incubation may be per-
exchanged weekly with fresh fluid before the additional
formed at ambient room temperature.
challenge. The percentage of fluid exchange should reflect
anticipated fluid turnover rates in fluid’s end-use application. 6.7 Manifold, aquarium style, multi-valve.
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.
E2275–03
NOTE 5—The number of manifolds and valves per manifold will
prepared end-use dilution, negative-control metalworking
depend on the number of microcosms in the test array. Air for each
fluid. Aerate at 25 6 2°C or at ambient room temperature for
microcosm shall be supplied through a single air valve. Where used, air 9
24 h or until the microbial viable count reaches 10 CFU ·
sterilization filters shall be placed between the air valve and microcosm
-1
mL . Replace 800 mL of this fluid with freshly prepared
aeration tube.
portion of the negative-control fluid. Repeat the aeration and
6.8 Metal Punch, 1 cm diameter.
metalworking fluid replacement procedure for a minimum of
three cycles before using the preparation as an inoculum.
7. Reagents and Materials
7.2.1.2 Prepare a characterized inoculum by using standard
7.1 Reagents:
microbiological techniques to isolate, maintain and identify
7.1.1 Purity of Reagents—Reagent grade chemicals shall be
specific microbes from spoiled metalworking fluid. Alterna-
used in all tests. Unless otherwise indicated, it is intended that
tively, cultures of specific interest may be obtained from a
all reagents conform to the specifications of the Committee on
commercial type culture collection. Examples of commercial
Analytical Reagents of the American Chemical Society where
cultures that may be used are: Aeromonas hydrophila
such specifications are available.
(ATCC 13444), Candida albicans (ATCC 752), Desulfovibrio
7.1.2 Water Purity—Unless otherwise indicated, references
desulfuricans (ATCC 7757), Escherichia coli (ATCC 8739),
to water shall be understood to mean reagent water as defined
Flavobacterium ferrugineum (ATCC 13524), Fusarium ox-
by Type III of Specification D 1193.
ysporum (ATCC 7601), Klebsiella pneumonia (ATCC 13883),
7.1.3 Antimicrobial Pesticide(s):
Mycobacterium immunogenum (Rossmoore strain), Proteus
mirabilis (ATCC 4675), Pseudomonas aeruginosa
NOTE 6—The measurement of antimicrobial pesticide (microbicide)
(ATCC 8689), Pseudomonas oleovorans (ATCC 8062) and
efficacy in a medium as complex as metalworking fluid is relative, not
Saccharomyces cerevisiae (ATCC 2338). Before using a char-
absolute. Consequently, when this method is used to evaluate microbicide
performance (8.3 or 8.4), it is prudent to always evaluate at least two
acterized inoculum for metalworking fluid bioresistance test-
antimicrobial treatments. Preferably one treatment should serve as a
ing, acclimate the inoculum following the procedure described
positive control; its efficacy in the test system having been established
for an uncharacterized inoculum (7.2.1.1). Warning— mi-
previously.
crobes recovered from metalworking fluids may be pathogenic.
7.1.4 Metalworking Fluid(s):
Do not pipet by mouth.
NOTE 7—The number of metalworking fluids available is almost
NOTE 9—As more bioresistant metalworking fluid formulations are
limitless. Recommendations for the use of any particular fluid cannot be
developed, microbicide-free control fluid may not support microbial
made. If the primary intent is to evaluate the general efficacy of the
growth at normal end-use dilutions. If microbial viable counts do not
4 -1
microbicide(s) being tested, then it/they should be tested in various types
increase by at least three logs within 48 h (for example, 10 CFU·mL
7 -1
of formulations. If the primary intent is to protect a particular formulation,
at time 0; 10 CFU·mL at time 48), then the coolant should be
then a microbicide-free version of that formulation should be used as the
augmented with 1 part in 10 of soybean-casein digest (7.1.3).
control and base-fluid to which the treatments are added.
7.2.2 Metal Chips:
7.1.4.1 End-use Dilution Metalworking Fluid—Dilute met-
NOTE 10—Although ferrous chips are suitable for most tests, alternative
alworking fluid concentrate in synthetic hard water
materials may be substituted if t
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