1、Designation: E1153 03 (Reapproved 2010)1E1153 14Standard Test Method forEfficacy of Sanitizers Recommended for InanimateInanimate, Hard, Nonporous Non-Food Contact Surfaces1This standard is issued under the fixed designation E1153; the number immediately following the designation indicates the year
2、oforiginal 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 NOTEA warning note was moved into the text editorially in May 2010.
3、1. Scope1.1 This test method is used to evaluate the antimicrobial efficacy of sanitizers on precleaned, inanimate, hard, nonporous,non-food contact surfaces.surfaces against Staphylococcus aureus, or Klebsiella pneumoniae or Enterobacter aerogenes, or acombination thereof.Appropriate modifications
4、to the method may be required when testing organisms not specified herein. Whenutilizing test surfaces not described herein (see Test Method E2274) or when evaluating spray-based or towelette-basedantimicrobial products, modifications may also be required.1.2 This test method may also be used to eva
5、luate the antimicrobial efficacy of one-step cleaner/sanitizercleaner-sanitizerformulations recommended for use on lightly soiled, inanimate, nonporous, non-food contact surfaces.1.3 It is the responsibility of the investigator to determine whether Good Laboratory Practices (GLP) isare required and
6、tofollow them where appropriate (see section 40 CFR, 160)160 or as revised.revised.)1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.1.5 This standard may involve hazardous materials, chemicals and microorganisms and should
7、 be performed only by personswho have had formal microbiological training.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 anddetermine the
8、applicability of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D1193 Specification for Reagent WaterE1054 Test Methods for Evaluation of Inactivators of Antimicrobial AgentsE2274 Test Method for Evaluation of Laundry Sanitizers and DisinfectantsE2756 Terminology Rela
9、ting to Antimicrobial and Antiviral Agents2.2 Federal Standard:40 CFR, Part 160, Good Laboratory Practice Standards33. Terminology3.1 Terms used in this test method are defined in Terminology E2756.3.2 Definitions of Terms Specific to This Standard:3.2.1 accuracy, na measure of the degree of conform
10、ity of a value generated by a specific procedure to the assumed oraccepted true value, and includes both precision and bias.3.2.2 ambient temperature, ntemperature of the environment in which a test method is performed.3.2.3 antimicrobial, adjdescribes an agent that kills or inactivates microorganis
11、ms or suppresses their growth or reproduction.1 This test method 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 1, 2010April 1,
12、 2014. Published May 2010May 2014. Originally approved 1987. Last previous edition approved in 20032010 asE1153 03.E1153 03(2010)1. DOI: 10.1520/E1153-03R10.10.1520/E1153-14.2 For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at serviceastm.org. For A
13、nnual Book of ASTM Standardsvolume information, refer to the standards Document Summary page on the ASTM website.3 Available from the Superintendent of Documents, U.S. Government Printing Office, Washington, DC 20402.This document is not an ASTM standard and is intended only to provide the user of a
14、n ASTM 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 publi
15、shed 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 States13.2.4 bias, na systematic error that contributes to the difference between the mean of a large number of test results and anaccepte
16、d reference value.3.2.5 cleaner-sanitizer, na physical or chemical agent that removes soil from an object and reduces numbers ofmicroorganisms on non-food contact surfaces.3.2.6 carrier, na surrogate surface or matrix that facilitates the interaction of test microorganisms and treatment(s).3.2.7 eff
17、cacy, nthe proven performance of a product established under defined conditions of testing.3.2.8 inoculum, nthe viable microorganisms used to contaminate a sample, device or surface, often expressed as to numberand type.3.2.9 neutralization, nthe process for inactivating or quenching the activity of
18、 a microbiocide, often achieved through physical(for example, filtration or dilution) or chemical means.3.2.10 precision, nthe closeness of agreement between independent test results obtained under prescribed conditions.3.2.11 reproducibility, nthe precision of test results obtained in different lab
19、oratories performing the same test procedure underspecifically defined conditions.3.2.12 sanitizer, nchemical or physical agent(s) used to reduce the number of microorganisms to a level judged to beappropriate for a defined purpose and/or claim.4. Significance and Use4.1 This test method shall be us
20、ed to determine if a chemical has application intended for use as a non-food contact sanitizeror as a one-step cleaner/sanitizer.cleaner-sanitizer provides percent reductions of the selected test organisms on treated carriers ascompared to control.5. Apparatus5.1 BalanceAcalibrated balance with a pl
21、atform to accommodate a 100-mLvolumetric flask.This balance should be sensitiveto 0.01 g.5.2 Nonporous Test Surfaces, pre-cleaned.5.2.1 Borosilicate Glass Squares, 25 by 25 by 2 mm slides, nonchipped.or 18 mm by 36 mm slides, nonchipped. 3 in. by 1 in.(76 mm by 25 mm) nonchipped slides may be used f
22、or towelette applications5.2.2 Glazed Glass or Stainless Steel, of appropriate type, approximately same size as in 4.2.15.2.1.5.3 Glass Culture Tubes, recommended sizes: 18 to 20 by 150 mm and 25 by 150 mm without lip.5.4 Culture Tube Closures, appropriate sized nontoxic closures.5.5 Pipets or Dispe
23、nsing Syringes, (or both), appropriately calibrated and sterile.5.6 Bacteriological Transfer Loop, 4 mm inside diameter loop of platinum or platinum alloy wire or sterile, disposable plasticloops of approximatesame size.5.7 Flasks or Containers:5.7.1 Appropriate sizes with closures for preparation o
24、f culture medium and sterile distilleddeionized water.5.7.2 Volumetric, 100 and 1000 mL, sterile.5.8 Petri dishes, recommended sizes: 50 by 9 mm plastic, and 100 by 15 mm, glass and plastic; sterile.5.9 Jars, ointment jars, (for example polypropylene) 2 oz (60 mL) mL), recommended, with nontoxic lid
25、s, sterile.5.10 Graduated Cylinders, recommended sizes; 100 and 500 mL.5.11 Flaming ApparatusA bunsen burner or other appropriate heat sterilizer.5.12 MixerA “vortex” mixer is recommended.5.13 TimerA reliable stopwatch or laboratory timer capable of measuring elapsed time in seconds and minutes.5.14
26、 pH MeterA reliable reliable, standardized pH meter to determine pH of culture media.5.15 Desiccator, recommended size: 200 mm inside diameter with approximately 125-mm chamber depth from inside plate tocover flange, glass.5.16 Incubator, capable of maintaining temperature of 37 6 2C.25 to 32C or 35
27、 to 39C, or both.5.17 Sterilizer, steam sterilizer and hot air oven (180(180 6 2C for 22 h).5.18 Colony CounterAny one of several types may be used, for example Quebec.5.19 Membrane Filters, of 0.22 m pore size.Compatible with the test organism (for example, 0.45 m pore size).5.20 Filter Assembly, a
28、utoclavable.autoclavable or pre-sterilized.E1153 1425.21 Forceps. Forceps (may be autoclave sterilized prior to use).5.22 Refrigerator, capable of maintaining 2 to 8C.6. Reagents and Materials6.1 Purity of ReagentsReagent grade chemicals shall be used in all tests. Unless otherwise indicated, it is
29、intended that allreagents shall conform to the specifications of the Committee on Analytical Reagents of the American Chemical Society, wheresuch specifications are available.4 Other grades may be used, provided it is first ascertained that the reagent is of sufficiently highpurity to permit its use
30、 without lessening the accuracy of the determination.6.2 Water for Dilution of Product Under Test:6.2.1 Water, sterile, deionized or distilled, equivalent to or better than Type 3, see Specification D1193.6.2.2 Association of Offcial Analytical Chemists (AOAC) Synthetic Hard Water:5(c)6.2.2.1 Soluti
31、on 1Dissolve 31.74 g magnesium chloride (MgCl2) (or equivalent of hydrates) and 73.99 g calcium chloride(CaCl2) in boiled distilled or deionized water and dilute to 1 L. Sterilize by autoclaving.6.2.2.2 Solution 2Dissolve 56.03 g sodium bicarbonate (NaHCO3) in boiled distilled or deionized water and
32、 dilute to 1 L.Sterilize by membrane filtration.6.2.2.3 Place the desired amount of Solution 1 in a sterile 1-L volumetric flask, or other appropriate volumetric vessel. Each1 mL of Solution 1 will give a water equivalent to ca. 100 ppm of hardness calculated as calcium carbonate (CaCO3flask and add
33、) by the equation below. (For example, 4 mL of solution 1 would be added to the flask to target 400 ppm hardness in 1L of water.)Add approximately 600 mL or 34sterile distilled water; of the total water volume of sterile distilled or deionized (reagent grade)water free of substances that interfere w
34、ith analytical methods; then add 4 mL of Solution 2 and dilute to exactly 1 L with steriledistilled water. Each millilitre of Solution 1 will give a water equivalent to 100 ppm of hardness calculated as calcium carbonate(CaCOor deionized water. 3) by the following equation:Total hardness as ppm CaCO
35、3 (1)52.4953ppm Ca#14.1153ppm Mg#6.2.3 The final pH of synthetic hard water should be from 7.6 to 8.2.8.0.6.2.4 The synthetic water to be used for the testing should be analyzed chemically for hardness at the time of test.Analysis maybe performed by the method described in footnote 6(5(c) or by comm
36、ercial available mercially available kit. The watermust be used within 24 h of preparation but may be refrigerated at 2 to 8C prior to use.The solution must be analyzed for hardnesson the day of use.6.2.5 All water used for preparation of test solutions shall be sterile.6.3 Sanitizing SolutionsFresh
37、ly prepared solutions of sanitizers (for example, used within 8 h of dilution) shall be used in alltests.6.4 Neutralizing SolutionsSolutions appropriate to inactivate sanitizing solutions shall be used in accordance with PracticesE1054.6.5 Culture Media:56.5.1 Nutrient Broth.(5(a)6.5.2 Nutrient Agar
38、.(5(b)6.5.3 Tryptic Soy Broth, per manufacturers instructions6.5.4 Other appropriate growth medium or subculture agar may be used where appropriate for the test organism (prepared permanufacturers instructions or purchased commercially).6.6 Soil, Fetal Bovine Serum, aseptically derived and maintaine
39、d.7. Preparation of Apparatus7.1 Constant Humidity Chamber (Desiccator):7.1.1 At least one day prior to use, fill the lower portion of a large size desiccator with about 500 mLof glycerin solution havinga refractive index of 1.4529 at 25C (approximately 86.5 % glycerin in distilled water will provid
40、e this refractive index). This willprovide a constant 40 to 41 % relative humidity at 37 6 2C35 to 39C in which the inoculated nonporous square surfaces will4 Reagent Chemicals, American Chemical Society Specifications, American Chemical Society, Washington, DC. For suggestions on the testing of rea
41、gents not listed bythe American Chemical Society, see Analar Standards for Laboratory Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia and NationalFormulary, U.S. Pharmaceutical Convention, Inc. (USPC), Rockville, MD.5 “Official Methods ofAnalysis of theAssociation of Off
42、icialAnalytical Chemists,”Association of OfficialAnalytical Chemists, Washington, DC, Chapter 6: Disinfectants,15th ed., 1990.6.(a) Page 133,Method 955.11 Section 955.11 A. (a).(b) Page 133,Method 955.11 Section 955.11 A. (c).(c) Page 139140, Section 960.09A.Method 960.09 Section Sections D and E.E1
43、153 143be dried prior to treatment with the sanitizer. Replace the porcelain floor plate of the desiccator and store at 37 6 2C35 to 39Cto allow to come to equilibrium. Alternatively, a humidity controlled incubator set to 35 to 39C may be used to achieve dryingconditions appropriate for maximum sur
44、vival of the test organism.7.2 Test Squares:7.2.1 Test squares shall be dipped in acetone or 70 to 95 % ethyl or isopropyl alcohol, rinsed with distilled or deionized water,and air dried before sterilization.7.2.2 Place test squares into a large, glass petri dish and sterilize in a hot air oven for
45、22 h at 180C.180C.7.2.3 After sterilization, place each square into separate 50 by 9 mm or 100 by 15 mm sterile plastic petriPetri dishes usingsterile technique.8. Test Organisms8.1 Klebsiella (K.) pneumoniae American Type Culture Collection (ATCC) 4352 or Enterobacter aerogenes American TypeCulture
46、 Collection (ATCC) 13048 and Staphylococcus (S.) aureus ATCC 6538.8.2 Maintenance of Test OrganismsMaintain stock cultures of on nutrient agar. Incubate 2 days at 35 to 39C for K.pneumoniae and S. aureus on nutrient agar. Incubate 2or 25 to 32C for days atE. aerogenes, 37 6 2C, then refrigerate at 5
47、 to7C. Transfer culture every 3 days. approximately 2 to 8C for up to one month (for example, up to 31 days). To prepare the testinocula, transfer each culture for at least 3 days (transfers) as described in 9.1. Stock slant cultures used for inoculation should notbe more than five passages removed
48、from the ATCC cultures (USP XXIII).6 Information on long term culture maintenance andstorage is found in “Manual of Methods for General Bacteriology”7 and “ATCC Catalogue of Bacteria and Bacteriophages”.89. Preparation of Inocula9.1 K. pneumoniae and S. aureusK. pneumoniae K. pneumoniae and S. aureu
49、s are grown in nutrient broth. E. aerogenes isgrown in tryptic soy broth. From stock cultures, (no more than 30 days1 month old), inoculate tubes containing 10 mL ofappropriate broth, and incubate for 24 h at 37 6 2C. 6 2h at 35 to 39C for K. pneumoniae and S. aureus or 25 to 32C for E.aerogenes. Using a 4 mm inside diameter transfer loop, transfer a loopfullloopful of the culture into fresh broth. Make at least threeconsecutive daily transfers prior to use as an inoculum. The final transfer is incubated
