1、Designation: E2275 03 (Reapproved 2008)E2275 13Standard Practice forEvaluating Water-Miscible Metalworking Fluid Bioresistanceand Antimicrobial Pesticide Performance1This standard is issued under the fixed designation E2275; the number immediately following the designation indicates the year oforigi
2、nal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.1. Scope1.1 This practice addresses the evaluation of the relative inherent
3、bioresistance of water-miscible metalworking fluids, thebioresistance attributable to augmentation with antimicrobial pesticides or both. It replaces Methods D3946 and E686.1.2 In this practice relative bioresistance is determined by challenging metalworking fluids with a biological inoculum that ma
4、yeither be characterized (comprised of one or more known biological cultures) or uncharacterized (comprised of biologicallycontaminated metalworking fluid or one or more unidentified isolates from deteriorated metalworking fluid). Challenged fluidbioresistance is defined in terms of resistance to bi
5、omass 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 orend-use dilution. It is also applicable to metal
6、working fluids that are formulated using non-microbicidal, inherently bioresistantcomponents.1.4 The values stated in SI units are to be regarded as the standard. The values given in parentheses are for information only.1.5 This standard does not purport to address all of the safety concerns, if any
7、 associated with its use. It is the responsibilityof the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatorylimitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D1129 Terminology Relating to WaterD888 Test Methods f
8、or Dissolved Oxygen in WaterD1067 Test Methods for Acidity or Alkalinity of WaterD1193 Specification for Reagent WaterD3342 Test Method for Dispersion Stability of New (Unused) Rolling Oil Dispersions in WaterD3519 Test Method for Foam in Aqueous Media (Blender Test) (Withdrawn 2013)3D3601 Test Meth
9、od for Foam In Aqueous Media (Bottle Test) (Withdrawn 2013)3D4012 Test Method for Adenosine Triphosphate (ATP) Content of Microorganisms in WaterD4627 Test Method for Iron Chip Corrosion for WaterMiscible Metalworking FluidsD5465 Practice for Determining Microbial Colony Counts from Waters Analyzed
10、by Plating MethodsE70 Test Method for pH of Aqueous Solutions With the Glass ElectrodeE1326 Guide for Evaluating Nonconventional Microbiological Tests Used for Enumerating BacteriaE2169 Practice for Selecting Antimicrobial Pesticides for Use in Water-Miscible Metalworking FluidsE2523 Terminology for
11、 Metalworking Fluids and OperationsE2563 Practice for Enumeration of Non-Tuberculosis Mycobacteria in Aqueous Metalworking Fluids by Plate Count MethodE2564 Practice for Enumeration of Mycobacteria in Metalworking Fluids by Direct Microscopic Counting (DMC) MethodE2657 Test Method for Determination
12、of Endotoxin Concentrations in Water-Miscible Metalworking Fluids1 This practice is under the jurisdiction of ASTM Committee E35 on Pesticides, Antimicrobials, and Alternative Control Agents and is the direct responsibility ofSubcommittee E35.15 on Antimicrobial Agents.Current edition approved April
13、 1, 2008Oct. 1, 2013. Published May 2008November 2013. Originally approved in 2003. Last previous edition approved in 20032008 asE2275 031.(2008). DOI: 10.1520/E2275-03R08.10.1520/E2275-13.2 For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at service
14、astm.org. For Annual Book of ASTM Standardsvolume information, refer to the standards Document Summary page on the ASTM website.3 The last approved version of this historical standard is referenced on www.astm.org.This document is not an ASTM standard and is intended only to provide the user of an A
15、STM standard an indication of what changes have been made to the previous version. Becauseit may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current versionof the standard as publishe
16、d by ASTM is to be considered the official document.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States1E2694 Test Method for Measurement of Adenosine Triphosphate in Water-Miscible Metalworking FluidsE2756 Terminology Relating to Antimic
17、robial and Antiviral AgentsE2889 Practice for Control of Respiratory Hazards in the Metal Removal Fluid Environment2.2 Other Standards:4.027 Synthetic Hard Water49215A.6a Heterotrophic Plate Count Media, Plate Count Agar59216 Direct Total Microbial Count5Microbiological Test 62.3 Government Standard
18、40 CFR 156 Labeling Requirements for Pesticides and Devices3. Terminology3.1 For definitions of terms used in this guide refer to Terminologies D1129, E2523, and E2756.3.2 Definitions:3.2.1 active ingredient, nthe chemical component or components of an antimicrobial pesticide that provides its micr
19、obicidalperformance.3.2.2 antimicrobial pesticide, nchemical additive registered under 40 CFR 152, for use to inhibit growth, proliferation or bothof microorganisms.3.2.3 as supplied, adjantimicrobial pesticide finished product including the active ingredient(s), solvent and any additionalinactive i
20、ngredients.3.2.4 biocide, nany chemical intended for use to kill organisms.3.2.5 bioresistant, adjability to withstand biological attack.3.2.5.1 DiscussionBioresistant, or recalcitrant, chemicals are not readily metabolized by microorganisms.3.2.6 biostatic, adjable to prevent existing microbial con
21、taminants from growing or proliferating, but unable to kill them.3.2.6.1 DiscussionBiostatic additives may be registered antimicrobial pesticides or unregistered chemicals with other performance properties. Thedifference between biocidal and biostatic performance may be attributed to dose, chemistry
22、 or both.3.2.7 dose, nconcentration of antimicrobial pesticide added to treated solution.3.2.7.1 DiscussionDose is generally expressed as either ppm active ingredient (a.i.) or ppm as supplied (a.s.).3.2.8 inactive ingredient, ncomponent of antimicrobial pesticide that is not directly responsible fo
23、r the pesticidesantimicrobial performance.3.2.8.1 DiscussionInactive ingredients may include, but are not limited to solvents and chemicals that improve the pesticides non-biocidalperformance properties, such as miscibility and reactivity with non-target molecules in the treated material.3.2.9 minim
24、um inhibitory concentration (MIC), nlowest treatment-dose that will prevent test population from growing,proliferating or otherwise contributing to biodeterioration.3.3 Abbreviations:3.3.1 a.i.active ingredient3.3.2 a.s.as supplied3.3.3 ATCCamerican type culture collection3.3.4 CFUcolony forming uni
25、t4 AOAC International Methods of Analysis, AOAC International, Gaithersburg, MD.5 Available fromAmerican Public HealthAssociation (APHA) Standard Methods for the Examination of Water and Wastewater 800 I Street, NW Washington, DC 20001.6 Available from U.S. Pharmacopeia (USP), 12601 Twinbrook Pkwy.,
26、 Rockville, MD 20852-1790, http:/www.usp.org.E2275 1324. Summary of Practice4.1 End-use dilutions of one or more water-miscible metalworking fluids are dispensed into microcosms. The fluids may befresh or aged, dosed with one or more antimicrobial pesticides or undosed. Microcosms are challenged wit
27、h either uncharacterizedor characterized biological inocula. After inoculation, microcosms are aerated either continuously or periodically to simulaterecirculation conditions in coolant systems. Chips may also be added to microcosms to simulate chip accumulation in coolantsystems.4.2 After inoculati
28、on, fluid samples are drawn from each microcosm periodically and tested for the parameters of interest,including but not limited to microbial viable counts. Depending on the test objectives, the test duration may range from 24 h tothree months.4.2.1 Shorter test periods are used to evaluate microbic
29、ide speed of kill and metalworking formulation initial bioresistance.4.2.2 Longer test periods are used to evaluate metalworking fluid formulation resistance to repeated challenges. For tests lastinglonger than one-week, 10 to 80 % of the fluid is exchanged weekly with fresh fluid before the additio
30、nal challenge. The percentageof fluid exchange should reflect anticipated fluid turnover rates in fluids end-use application.4.3 Bioresistance is determined as the test fluids relative ability to prevent the proliferation of challenge microbes, retain itsoriginal chemical or physical properties of s
31、ome combination of the above.The bioresistance of test formulations is defined relativeto that of a benchmark or control formulation.5. Significance and Use5.1 This practice provides laboratory procedures for rating the relative bioresistance of metalworking fluid formulations, fordetermining the ne
32、ed for microbicide addition prior to or during fluid use in metalworking systems and for evaluating microbicideperformance. General considerations for microbicide selection are provided in Practice E2169.5.2 The factors affecting challenge population numbers, taxonomic diversity, physiological state
33、 inoculation frequency andbiodeterioration effects in recirculating metalworking fluid systems are varied and only partially understood. Consequently, theresults of tests completed in accordance with this practice should be used only to compare the relative performance of productsor microbicide tre
34、atments included in a test series. Results should not be construed as predicting actual field performance.6. Apparatus6.1 Air Supply, air provided at no more than 110 kPa.NOTE 1Any air source that is free of organic vapors, organic matter or other objectionable material may be used. Sterile air need
35、 not be used forthe uncharacterized inoculum, but shall be used for the characterized inoculum. If necessary, air may be sterilized either by inserting, in series, twocommercially available in-line sterile filters designed for this purpose. Alternatively an in-line filter may be prepared as follows:
36、 Pack two 150 mm longdrying tubes (bulb-type) loosely with borosilicate glass wool in series with neoprene stoppers, glass tubing and neoprene tubing. Wrap loosely inaluminum foil and steam sterilize at 103 to 138 kPa (15 to 20 psi) for 30 min or dry heat sterilize at 160C for 2 h. Cool to room temp
37、erature whilewrapped. Insert into air line with bulbs on upstream side. Whether using a commercial or fabricated filter, average lifetime in continuous use is two weeks.Discard sooner if upstream filter becomes wet or contaminated with oil.6.2 Aquarium Tubing, 6.35 mm (0.25 in.) diameter, silicone o
38、r vinyl.6.3 Autoclave, with both steam cycle (80 to 100C) and sterilization cycle (15 min at 121C) capability.6.4 Adjustable Volume Pipetters, with sterile disposable tips. Pipetters will be used to deliver 1.0 L to 2 mL volumes.6.5 Glassware:NOTE 2Sterile laboratory ware or sterile disposable labor
39、atory ware should be used according to standard microbiological practice.6.5.1 Glass Tubing, 6.35 mm (0.25 in.) i.d., cut into 15 cm lengths with ends fire-polished.6.5.2 French Square Bottles, 960 mL, with metal cap.NOTE 3Alternatively, 1 L capacity canning jars may be used.6.5.3 Pipetes, Bacteriol
40、ogical, 10 and 2.2 mL.6.6 Incubator, capable of maintaining a temperature of 25 6 2C.NOTE 4Although an incubator is preferred, incubation may be performed at ambient room temperature.6.7 Manifold, aquarium style, multi-valve.NOTE 5The number of manifolds and valves per manifold will depend on the nu
41、mber of microcosms in the test array. Air for each microcosm shallbe supplied through a single air valve. Where used, air sterilization filters shall be placed between the air valve and microcosm aeration tube.6.8 Metal Punch, 1 cm diameter.7. Reagents and Materials7.1 Reagents:E2275 1337.1.1 Purity
42、 of ReagentsReagent grade chemicals shall be used in all tests. Unless otherwise indicated, it is intended that allreagents conform to the specifications of the Committee on Analytical Reagents of the American Chemical Society where suchspecifications are available.77.1.2 Water PurityUnless otherwis
43、e indicated, references to water shall be understood to mean reagent water as defined byType III of Specification D1193.7.1.3 Antimicrobial Pesticide(s):NOTE 6The measurement of antimicrobial pesticide (microbicide) efficacy in a medium as complex as metalworking fluid is relative, not absolute.Cons
44、equently, when this method is used to evaluate microbicide performance (8.3 or 8.4), it is prudent to always evaluate at least two antimicrobialtreatments. Preferably one treatment should serve as a positive control; its efficacy in the test system having been established previously.7.1.4 Metalworki
45、ng Fluid(s):NOTE 7The number of metalworking fluids available is almost limitless. Recommendations for the use of any particular fluid cannot be made. If theprimary intent is to evaluate the general efficacy of the microbicide(s) being tested, then it/they should be tested in various types of formul
46、ations. If theprimary intent is to protect a particular formulation, then a microbicide-free version of that formulation should be used as the control and base-fluid towhich the treatments are added.7.1.4.1 End-use Dilution Metalworking FluidDilute metalworking fluid concentrate in synthetic hard wa
47、ter (AOAC 4.027) toachieve the concentration at which it is used typically in recirculating metalworking fluid systems.NOTE 8Depending on the metalworking process, metal alloy being worked and formulation chemistry, metalworking fluid end-use dilution may rangefrom 2 % (vv) to 15 % (vv). If the form
48、ulation(s) being tested is (are) likely to be used at a variety of end-use strengths, they should be tested minimallyat the high and low ends of the anticipated end-use concentration range. If the test objective is to evaluate microbicide performance in multiplemetalworking fluid formulations, a 5 %
49、 (vv) end-use dilution is appropriate.7.2 Materials:7.2.1 InoculumThe microbial inoculum may vary according to the users requirements. It may be either characterized oruncharacterized. The challenge population should be acclimated to the metalworking fluid before being used in this method.Acclimatization shall be achieved by growing the challenge in the end-use dilution, negative-control metalworking fluidformulation.7.2.1.1 Prepare an uncharacterized inoculum by adding 50 mL of spoiled metalworking fluid to 850 mL of freshly prepar
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