ASTM E1153-2003(2010)e1 Standard Test Method for Efficacy of Sanitizers Recommended for Inanimate Non-Food Contact Surfaces《无机非食品接触面用推荐的消毒器效力的标准试验方法》.pdf

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1、Designation: E1153 03 (Reapproved 2010)1Standard Test Method forEfficacy of Sanitizers Recommended for Inanimate Non-Food Contact Surfaces1This standard is issued under the fixed designation E1153; the number immediately following the designation indicates the year oforiginal adoption or, in the cas

2、e 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.1NOTEA warning note was moved into the text editorially in May 2010.1. Scope1.1 This test method is use

3、d to evaluate the antimicrobialefficacy of sanitizers on precleaned inanimate, nonporous,non-food contact surfaces.1.2 This test method may also be used to evaluate theantimicrobial efficacy of one-step cleaner/sanitizer formula-tions recommended for use on lightly soiled, inanimate,nonporous, non-f

4、ood contact surfaces.1.3 It is the responsibility of the investigator to determinewhether Good Laboratory Practices (GLP) is required and tofollow them where appropriate (see section 40 CFR, 160) or asrevised.1.4 The values stated in SI units are to be regarded asstandard. No other units of measurem

5、ent are included in thisstandard.1.5 This standard may involve hazardous materials, chemi-cals and microorganisms and should be performed only bypersons who have had formal microbiological training. Thisstandard does not purport to address all of the safety concerns,if any, associated with its use.

6、It is the responsibility of the userof this standard to establish appropriate safety and healthpractices and determine the applicability of regulatory limita-tions prior to use.2. Referenced Documents2.1 ASTM Standards:2D1193 Specification for Reagent WaterE1054 Test Methods for Evaluation of Inacti

7、vators of An-timicrobial Agents2.2 Federal Standard:40 CFR, Part 160, Good Laboratory Practice Standards33. Significance and Use3.1 This test method shall be used to determine if a chemicalhas application as a non-food contact sanitizer or as a one-stepcleaner/sanitizer.4. Apparatus4.1 BalanceA bala

8、nce with a platform to accommodate a100-mL volumetric flask. This balance should be sensitive to0.01 g.4.2 Nonporous Test Surfaces, pre-cleaned.4.2.1 Borosilicate Glass Squares, 25 by 25 by 2 mm slides,nonchipped.4.2.2 Glazed Glass or Stainless Steel, of appropriate type,approximately same size as i

9、n 4.2.1.4.3 Glass Culture Tubes, recommended sizes: 18 to 20 by150 mm and 25 by 150 mm without lip.4.4 Culture Tube Closures, appropriate sized nontoxic clo-sures.4.5 Pipets or Dispensing Syringes, (or both), appropriatelycalibrated and sterile.4.6 Bacteriological Transfer Loop, 4 mm inside diameter

10、loop of platinum or platinum alloy wire or sterile, disposableplastic loops of approximate size.4.7 Flasks or Containers:4.7.1 Appropriate sizes with closures for preparation ofculture medium and sterile distilled water.4.7.2 Volumetric, 100 and 1000 mL, sterile.4.8 Petri dishes, recommended sizes:

11、50 by 9 mm plastic,and 100 by 15 mm, glass and plastic; sterile.4.9 Jars, ointment jars, 2 oz (60 mL) with nontoxic lids,sterile.4.10 Graduated Cylinders, recommended sizes; 100 and500 mL.4.11 Flaming ApparatusA bunsen burner or other appro-priate heat sterilizer.1This test method is under the juris

12、diction of ASTM Committee E35 onPesticides and Alternative Control Agents and is the direct responsibility ofSubcommittee E35.15 on Antimicrobial Agents.Current edition approved April 1, 2010. Published May 2010. Originallyapproved 1987. Last previous edition approved in 2003 as E1153 03. DOI:10.152

13、0/E1153-03R10.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontact ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3Available from the Superintendent of Documents

14、, U.S. Government PrintingOffice, Washington, DC 20402.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.4.12 MixerA “vortex” mixer is recommended.4.13 TimerA reliable stopwatch or laboratory timer ca-pable of measuring elapsed time in

15、 seconds and minutes.4.14 pH MeterA reliable pH meter to determine pH ofculture media.4.15 Desiccator, recommended size: 200 mm inside diam-eter with approximately 125-mm chamber depth from insideplate to cover flange, glass.4.16 Incubator, capable of maintaining temperature of 37 62C.4.17 Sterilize

16、r, steam sterilizer and hot air oven (180 6 2Cfor 2 h).4.18 Colony CounterAny one of several types may beused, for example Quebec.4.19 Membrane Filters, of 0.22 m pore size.4.20 Filter Assembly, autoclavable.4.21 Forceps.5. Reagents and Materials5.1 Purity of ReagentsReagent grade chemicals shall be

17、used in all tests. Unless otherwise indicated, it is intended thatall reagents shall conform to the specifications of the Commit-tee on Analytical Reagents of the American Chemical Society,where such specifications are available.4Other grades may beused, provided it is first ascertained that the rea

18、gent is ofsufficiently high purity to permit its use without lessening theaccuracy of the determination.5.2 Water for Dilution of Product Under Test:5.2.1 Water, sterile, deionized or distilled, equivalent to orbetter than Type 3, see Specification D1193.5.2.2 Association of Offcial Analytical Chemi

19、sts (AOAC)Synthetic Hard Water:5(c)5.2.2.1 Solution 1Dissolve 31.74 g magnesium chloride(MgCl2) (or equivalent of hydrates) and 73.99 g calciumchloride (CaCl2) in boiled distilled water and dilute to 1 L.Sterilize by autoclaving.5.2.2.2 Solution 2Dissolve 56.03 g sodium bicarbonate(NaHCO3) in boiled

20、 distilled water and dilute to 1 L. Sterilizeby membrane filtration.5.2.2.3 Place the desired amount of Solution 1 in a sterile1-L flask and add approximately 600 mL sterile distilled water;then add 4 mL of Solution 2 and dilute to exactly 1 L withsterile distilled water. Each millilitre of Solution

21、 1 will give awater equivalent to 100 ppm of hardness calculated as calciumcarbonate (CaCO3) by the following equation:Total hardness as ppm CaCO3(1)5 2.495 3 ppm Ca# 1 4.115 3 ppm Mg#5.2.3 The pH of synthetic hard water should be from 7.6 to8.2.5.2.4 The synthetic water to be used for the testing s

22、houldbe analyzed chemically for hardness at the time of test.Analysis may be performed by the method described infootnote 6(c) or by commercial available kit.5.2.5 All water used for preparation of test solutions shall besterile.5.3 Sanitizing SolutionsFreshly prepared solutions ofsanitizers shall b

23、e used in all tests.5.4 Neutralizing SolutionsSolutions appropriate to inacti-vate sanitizing solutions shall be used in accordance withPractices E1054.5.5 Culture Media:55.5.1 Nutrient Broth.(5(a)5.5.2 Nutrient Agar.(5(b)5.6 Soil, Bovine Serum, aseptically derived and maintained.6. Preparation of A

24、pparatus6.1 Constant Humidity Chamber (Desiccator):6.1.1 At least one day prior to use, fill the lower portion ofa large size desiccator with about 500 mL of glycerin solutionhaving a refractive index of 1.4529 at 25C (approximately86.5 % glycerin in distilled water will provide this refractiveindex

25、). This will provide a constant 40 to 41 % relativehumidity at 37 6 2C in which the inoculated nonporoussquare surfaces will be dried prior to treatment with thesanitizer. Replace the porcelain floor plate of the desiccator andstore at 37 6 2C to allow to come to equilibrium.6.2 Test Squares:6.2.1 T

26、est squares shall be dipped in 70 to 95 % ethyl orisopropyl alcohol, rinsed with distilled water, and air driedbefore sterilization.6.2.2 Place test squares into a large, glass petri dish andsterilize in a hot air oven for2hat180C.6.2.3 After sterilization, place each square into separate 50by 9 mm

27、sterile plastic petri dishes using sterile technique.7. Test Organisms7.1 Klebsiella (K.) pneumoniae American Type CultureCollection (ATCC) 4352 and Staphylococcus (S.) aureusATCC 6538.7.2 Maintenance of Test OrganismsMaintain stock cul-tures of K. pneumoniae and S. aureus on nutrient agar. Incubate

28、2 days at 37 6 2C, then refrigerate at 5 to 7C. Transferculture every 3 days. Stock cultures used for inoculation shouldnot be more than five passages removed from the ATCCcultures (USP XXIII).6Information on long term culturemaintenance and storage is found in “Manual of Methods forGeneral Bacterio

29、logy”7and “ATCC Catalogue of Bacteria andBacteriophages”.84Reagent Chemicals, American Chemical Society Specifications, AmericanChemical Society, Washington, DC. For suggestions on the testing of reagents notlisted by the American Chemical Society, see Analar Standards for LaboratoryChemicals, BDH L

30、td., Poole, Dorset, U.K., and the United States Pharmacopeiaand National Formulary, U.S. Pharmaceutical Convention, Inc. (USPC), Rockville,MD.5“Official Methods of Analysis of the Association of Official AnalyticalChemists,”Association of OfficialAnalytical Chemists, Washington, DC, Chapter 6:Disinf

31、ectants, 15th ed., 1990.(a) Page 133, Section 955.11 A. (a).(b) Page 133, Section 955.11 A. (c).(c) Page 139140, Section 960.09A.6Sterility Tests (71), United States Pharmacopeia (USP) XXII.7Manual of Methods for General Bacteriology, 1981, P. Gerhardt (ed. in chief)ASM Microbiology, Washington, DC.

32、8Associated Concentrates, Inc., 32-60 61st St., Woodside, NY 11377.E1153 03 (2010)128. Preparation of Inocula8.1 K. pneumoniae and S. aureusK. pneumoniae and S.aureus are grown in nutrient broth. From stock cultures, (nomore than 30 days old), inoculate tubes containing 10 mL ofappropriate broth, an

33、d incubate for 24 h at 37 6 2C. Using a4 mm inside diameter transfer loop, transfer a loopfull of theculture into fresh broth. Make three consecutive daily transfersprior to use as an inoculum. The final transfer is incubated for48 h, and this culture is used for the test.8.2 Inocula for Testing San

34、itizers for Use on PrecleanedSurfacesThoroughly mix 48 h culture of test organism on“vortex” mixer, then allow the culture to settle for 15 min.Decant upper two thirds of this suspension and use this as theinoculum for testing sanitizers for use on precleaned surfaces.8.3 Inocula for Testing Formula

35、tions as One-Step Cleaner/Sanitizers or Sanitizers for Use on Lightly Soiled SurfacesThoroughly mix 48 h culture of test organism on “vortex”mixer, then allow the culture to settle for 15 min. Decant uppertwo thirds of this suspension and add bovine serum (forexample, 19 mL of a 48 h bacterial cultu

36、re and 1 mL bovineserum). Use this suspension now containing bovine serum at5 % concentration as the inoculum for testing one-step cleaner/sanitizers or sanitizers for use on lightly soiled surfaces.9. Preparation of Test Solutions9.1 Prepare the sanitizer in accordance with the manufac-turers recom

37、mended dilution. Dilutions for the test may bemade in sterile distilled water or in AOAC formula synthetichard water of any hardness desired (see 5.2).9.2 For each organism to be tested prepare 100 mL aliquotsof the test solution.10. Preparation of Neutralizer Solutions10.1 Quarternary and Phenolic

38、Solutions:10.1.1 Phosphate Buffer Stock Solution (0.25 M)Dissolve34.0 g of potassium phosphate, monobasic (KH2PO4)in500mL distilled water; adjust the pH to 7.2 with 1N NaOH anddilute to 1 L.10.1.2 Phosphate Buffer Dilution WaterAdd 1.25 mL of0.25 M phosphate buffer stock solution to 1 L water anddis

39、pense in 99 mL portions. Autoclave for 20 min at 121C.10.1.3 Neutralizer StockMix 40.0 g Azolectin,8280 mLpolysorbate 80, and 1.25 mL phosphate stock solution buffer(see 10.1.1). Adjust to pH 7.2 with 1N NaOH. Dilute to 1 Lwith distilled water. Dispense in suitable portions and sterilizefor 20 min a

40、t 121C.10.1.4 Neutralizer SolutionMix 62.5 mL of neutralizerstock (see 10.1.3), 6.25 mL of phosphate buffer stock solution(see 10.1.1), and 381.25 mL of distilled water. Dispense 20 mLportions into 25 by 150 mm culture tubes and sterilize for 20min at 121C.10.2 Halogen SanitizersNeuralizer Solutions

41、, Dissolve0.31 g of sodium thiosulfate and 0.30 mL of Triton X-100 in500 mL of distilled water. Dispense 20 mL portions into 25 by150 mm culture tubes and sterilize for 20 min at 121C.10.3 Other Sanitizing AgentsUse appropriate neutralizers(see Practices E1054).11. Procedures11.1 Inoculation of Test

42、 Squares:11.1.1 Inoculate each sterile glass or other nonporous sur-face (see 6.2.3) squares with exactly 0.02 mL of 48 h culture.Spread the inoculum to within 3 mm of the edges of the square.Prepare appropriate number of test squares, depending uponthe test and its parameters.11.1.2 Number each pla

43、te used in the order in which thesquares are inoculated. Place all plates containing the inocu-lated squares in the 37C constant humidity desiccator. Set thelids of the plates slightly ajar and close the desiccator lid.Allow the squares to remain at this temperature and humidityfor exactly 35 min. (

44、WarningBe very careful to remove thedesiccator lid only long enough to place the 50 by 9 mm plateson the porcelain floor plate and set their lids ajar.)11.2 Inoculum Count:11.2.1 Plate the appropriate dilutions of K. pneumoniae andS. aureus using nutrient agar. Incubate the organisms for 48 hat 37 6

45、 2C. Count the colonies to determine the number oforganisms per mL of culture and present at the start of the test.Cultures used for further testing must be kept at 5 to 7C for nomore than 8 h.11.2.2 Report inoculum count for the test organisms.11.3 Sanitizer or Cleaner/Sanitizer Treatment of Inocul

46、atedTest Squares:11.3.1 Transfer five squares to five sterile 2 oz (60 mL)ointment jars using sterile forceps. Be sure to resterilize theforceps between each transfer. (Dip in 70 to 95 % ethyl orisopropyl alcohol and burn off). Mark each jar with a numbercorresponding to that on the plate from which

47、 the square wastaken.11.3.2 At zero time on the timer, cover inoculated squareNo. 1 (the first one inoculated) with exactly 5 mL of the testsolution using a sterile 5 mL pipette. At exactly 1 min, coversquare No. 2 with 5 mL of the test solution. Treat square No.3 in a like manner at 2 min, square N

48、o. 4 at 3 min, and squareNo. 5 at 4 min.11.3.3 At exactly 5 min on the timer, add 20 mL ofappropriate neutralizer solution into jar No. 1 and rotate the jarvigorously on an even plane for approximately 50 rotations tosuspend the surviving organisms. At 6 min, add 20 mL ofneutralizer into jar No. 2 a

49、nd rotate as in No. 1. Continueaddition of neutralizer to jars No. 3, No. 4 and No. 5 at 1 minintervals, and rotate each in turn.11.3.4 Within 30 min after the addition of the neutralizer tothe test sanitizer or cleaner/sanitizer, plate in duplicate 1.0 and0.1 mL of the neutralizer solution from each of the five jarsusing standard pour plate techniques. Use nutrient agar for bothorganism types. Incubate for 48 h, K. pneumoniae and S.aureus at 37 6 2C. Count the number of colonies on theplates.11.4 Inoculation of Control SquaresAllo