ASTM E1174-2013 Standard Test Method for Evaluation of the Effectiveness of Health Care Personnel Handwash Formulations《评价医护人员洗手液配方有效性的标准试验方法》.pdf

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1、Designation: E1174 13Standard Test Method forEvaluation of the Effectiveness of Health Care PersonnelHandwash Formulations1This standard is issued under the fixed designation E1174; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, t

2、he 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 test method is designed to determine the effective-ness of antimicrobial handwashing agents for the re

3、duction oftransient microbial flora when used in a handwashing proce-dure.1.2 A knowledge of microbiological techniques is requiredfor these procedures.1.3 This test method may be used to evaluate topicalantimicrobial handwash formulations.1.4 Performance of this procedure requires the knowledgeof r

4、egulations pertaining to the protection of human subjects.21.5 The values stated in SI units are to be regarded asstandard; except for distance, in which case inches are used andmetric units follow in parentheses.1.6 This standard does not purport to address all of thesafety concerns, if any, associ

5、ated with its use. It is theresponsibility of the user of this standard to establish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use. For more specificprecautionary statements see 8.2.2. Referenced Documents2.1 ASTM Standards:3E1054 Te

6、st Methods for Evaluation of Inactivators of Anti-microbial AgentsE2755 Test Method for Determining the Bacteria-Eliminating Effectiveness of Hand Sanitizer FormulationsUsing Hands of Adults3. Terminology3.1 Definitions:3.1.1 active ingredient, na substance added to a formula-tion specifically for t

7、he inhibition or inactivation of microor-ganisms.3.1.2 cleansing wash, na non-antimicrobial wash intendedto remove gross soil or residues from the hands of the panelistsprior to the conduct of the study and as noted throughout thestudy. This may also be referred to as a cosmetic wash.3.1.3 healthcar

8、e personnel handwash, na cleanser orwaterless agent intended to reduce transient bacteria on thehands.3.1.4 neutralization, na process which results in quench-ing the antimicrobial activity of a test material. This may beachieved through dilution of the test material(s) to reduce theantimicrobial ac

9、tivity, or through the use of chemical agents,called neutralizers, to eliminate antibacterial activity.3.1.5 resident microorganisms, nmicroorganisms that liveand multiply on the skin, forming a permanent population.3.1.6 test formulation, na formulation which incorporatesantimicrobial ingredient(s)

10、.3.1.7 test organisman applied inoculum of an organismthat has characteristics which allow it to be readily identified.The test organism is used to simulate a transient topicalmicrobial contaminant. It may also be referred to as a markerorganism, bacterial simulant, or bacterial contaminant.3.1.8 tr

11、ansient microorganismsorganisms from the envi-ronment that contaminate but do not normally colonize theskin.4. Summary of Test Method4.1 This test method is conducted on a group of volunteerpanelists who have refrained from using topical antimicrobialformulations for at least one week prior to the i

12、nitiation of thetest.Activity of the test material is measured by comparing thenumber of test organisms recovered from artificially contami-nated hands after use of a handwashing formulation to thenumber recovered from contaminated hands not exposed to thetest formulation. The method describes speci

13、fic procedures tobe followed using Serratia marcescens as the test organism.The activity of the test material is measured following a singlewash and may be measured following multiple washes in asingle day using a neutralization recovery method.1This test method is under the jurisdiction of ASTM Com

14、mittee E35 onPesticides, Antimicrobials, and Alternative Control Agents and is the directresponsibility of Subcommittee E35.15Antimicrobial Agents.Current edition approved Feb. 15, 2013. Published February 2013. Originallyapproved in 1987. Last previous edition approved in 2006 as E1174 06. DOI:10.1

15、520/E1174-13.2Federal Register, Vol 46, No. 17, Jan. 27, 1991.3For 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

16、.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States14.2 An alternative test organism is Escherichia coli. Culturemedia and incubation conditions appropriate for this organismshould be employed.The investigator should also be aware thatth

17、ere may be health risks associated with the use of thisorganism and precautions similar to those referenced in 8.2should be undertaken.5. Significance and Use5.1 The procedure may be used to test the effectiveness ofantimicrobial handwashing agents. The test formulations maybe designed for frequent

18、use to reduce the transient bacterialflora on hands. Alcohol-based hand rubs and other leave-onformulations used without the aid of water may be tested usingTest Method E2755.6. Apparatus6.1 Colony CounterAny of several types may be used, forexample, Quebec Colony Counter.6.2 IncubatorAny incubator

19、capable of maintaining thefollowing temperatures: S. marcescens (25 6 2C) or E. coli(35 6 2C). This temperature is required to ensure pigmentproduction for S. marcescens.6.3 SterilizerAny suitable steam sterilizer capable of pro-ducing the conditions of sterilization is acceptable.6.4 Timer (Stop-cl

20、ock)One that can be read for minutesand seconds.6.5 Handwashing SinkA sink of sufficient size to permitpanelists to wash without touching hands to sink surface orother panelists.6.5.1 Water Faucet(s)To be located above the sink at aheight which permits the hands to be held higher than theelbow durin

21、g the washing procedure.6.6 Tap Water Temperature Regulator and TemperatureMonitorTo monitor and regulate water temperature of 40 62C.7. Reagents and Materials7.1 Bacteriological Pipettes10.0 and 2.2-mL or 1.1-mLcapacity.NOTE 1Presterilized/disposable bacteriological pipettes are availablefrom most

22、local laboratory supply houses.7.2 Water Dilution BottlesAny sterilizable glass containerhaving a 150 to 200 mL capacity and tight closures may beused.NOTE 2Milk dilution bottles of 160-mL capacity having a screw-capclosure are available from most local laboratory supply houses.7.3 Erlenmeyer Flask2

23、-L capacity for culturing test or-ganism.7.4 Cleansing WashA mild, non-antimicrobial soft soap.Soft Soap, 200 g/LLinseed oil 50 parts by weightPotassium hydroxide 9.5 partsEthanol 7 partsDistilled or high purity water as needed7.4.1 Add linseed oil to a solution of potassium hydroxidein 15 parts wat

24、er and heat up to approximately 70C whileconstantly stirring.Add the ethanol and continue heating whilestirring until the saponification process is completed and asample dissolves clearly in water and almost clearly in alcohol.The weight of the soft soap is then brought up to 100 parts byaddition of

25、 hot water. Take 200 g of the soft soap in 1 L ofwater. Dispense in to appropriate containers and sterilize in anautoclave.7.5 Test MaterialDirections for use of the test materialmay be utilized. If directions are not available, use directionsprovided in this test method.7.6 GlovesLoose-fitting, unl

26、ined, powder-free gloveswhich possess no antimicrobial properties, or equivalent.4(Plastic bags with low bioburden may be used in place ofgloves.)7.7 Sampling SolutionDissolve 0.4 g KH2PO4, 10.1 gNa2HPO4, and 1.0 g isooctylphenoxypolyethoxyethanol andwith appropriately validated neutralizers in 1 L

27、distilled water.Adjust pH to 7.8 with 0.1 N HCl or 0.1 N NaOH. Dispense sothat final volume after sterilization is 75 mL, sterilized at121C.57.8 Dilution FluidSterile Butterfields Buffer6or othersuitable diluent, adjusted to pH 7.2 with effective neutralizerfor the test material.Adjust pH with 0.1 N

28、 HCl or 0.1 N NaOH.See Test Methods E1054.7.9 AgarSoybean-casein digest agar or other solid mediaappropriately validated to support growth of the test organismwith appropriate neutralizers if needed.7.10 BrothSoybean-casein digests broth or other liquidmedia appropriate to support growth of the test

29、 organism.8. Test Organism8.1 Serratia marcescens (ATCC 14756) is to be used as thetest organism. This is a strain having stable pigmentation at25C.8.2 Escherichia coli (ATCC 11229) are an alternative testorganism. When E. coli is used, the plating agar should includea suitable indicator (for exampl

30、e, MUG7). (WarningTheapplication of microorganisms to the skin may involve a healthrisk. Prior to applying the test organism to the skin, theantibiotic susceptibility profile of the strain should be deter-mined. If the strain is not susceptible to gentamicin, do not useit. If an infection occurs, th

31、e antibiotic sensitivity profile shouldbe made available to the attending clinician. Following the4A zone of inhibition test such as AATCC Test Method 90-1965 may be used toevaluate antimicrobial properties of gloves, AATCC Test Methods, AmericanAssociation ofTextile Chemists and Colorist, 1968Techn

32、ical Manual, Section B-75.5Peterson, A. F., “The Microbiology of the Hands: Evaluating the Effects of theSurgical Scrubs,” Developments in Industrial Microbiology, Vol 14, 1973, pp.125130.6Horowitz, W., (Ed.), Offcial Methods of Analysis of the AOAC, 17th Ed., Sec.6.3.03 A.(f), Chapter 6, 2000, p. 1

33、0. Official Methods of Analysis of AOACInternational, Gaitherburg, MD.7United States Pharmacopeia 28: United States Pharmacopeial Convention,Inc., Rockville, MD, Chapter entitled “Microbial Limits Test.” The MUG (4-methylumbelliferyl-D-gluconride) substrate is hydrolyzed by -D-gluconridase toyield a

34、 fluorescent end product, 4-methylumbelliferone. -D-gluconridase is pos-sessed by E. coli (ATCC 11229). MUG is incorporated into the appropriate growthmedium at 0.05 g/L.E1174 132subjects last contamination and wash with the formulation, thesubjects hands are to be sanitized by scrubbing with 70%iso

35、propanol solution or equivalent. The purpose of this alcoholscrub is to destroy residual test organisms on the skin.)8.3 Preparation of Test Organism Suspension8.3.1 S. marcescensA homogeneous culture is used toinoculate the hands. The stock culture, frozen or lyopholized,should be at least two 24-h

36、 soybean-casein digest broth (7.10)transfers from the original ATCC culture, but there should beno more than four transfers removed from the ATCC culture.From the stock culture of Serratia marcescens (ATCC 14756),inoculate the appropriate volume of soybean-casein digestbroth (7.10) with 0.1 mL of st

37、ock culture of S. marcescens/100mL of broth to yield the volume necessary to complete thestudy. Incubate for 24 6 4hat256 2C. Broth should developa red pigment.8.3.2 E. coliA homogeneous culture is used to inoculatethe hands. The stock culture should be at least two 24-hourbroth transfers from the o

38、riginal ATCC culture, but no morethan five transfers removed from the ATCC culture. From thestock culture of Escherichia coli (ATCC 11229), inoculate theappropriate volume of soybean-casein digest broth (7.10) with0.1 mL of stock culture/100 mL of broth to yield the volumenecessary to complete the s

39、tudy. Incubate for 24 6 4hat3562C.8.4 Swirl or shake suspension before the withdrawal of eachaliquot. Assay the suspension for number of organisms at thebeginning and end of the use period. Do not use a suspensionfor more than 8 h. The suspension may not vary more than60.5 log10cfu/mL over an 8 h pe

40、riod.9. Subjects9.1 Recruit a sufficient number of healthy adult humanvolunteers who have no clinical evidence of dermatoses, openwounds, hangnails, or other skin disorders.9.2 Instruct subjects to avoid contact with antimicrobialproducts (other than the test material as dispensed for each testwash)

41、 for the duration of the test and for at least one week priorto the test. This restriction includes antimicrobial-containingantiperspirants, deodorants, shampoos, lotions, and soaps, alsosuch materials as acids, bases, and solvents. Bathing in biocidetreated pools, hot tubs, or spas should be avoide

42、d. Subjects areto be provided with a kit of nonantimicrobial personal careproducts for exclusive use during the test and rubber gloves tobe worn when contact with antimicrobial or harsh chemicalscannot be avoided.10. Procedure10.1 After subjects have refrained from using antimicrobialformulations fo

43、r at least 7 days, they perform a 30 s cleansingwash (7.4) in the same manner that is described for the test andcontrol formulations. This procedure removes oil and dirt andfamiliarizes the panelists with the washing technique. For thisand all other washes and rinses, the water temperature isadjuste

44、d to 40 6 2C and the water flow rate to 4 Lper minute.This may be accomplished by placing a 2000 mL glass beakeror flask under each spigot to be used for subjects handwashing. Allow the water to flow into the beaker. Adjust thewater flow at each spigot accordingly, so that the beaker fillswithin 30

45、s.10.2 Hand ContaminationA liquid suspension of the testorganism containing between 5.0 108and 1.0 109cfu/mLis used. See Table 1.10.2.1 A 1.5 mL aliquot of the test organism suspension isdispensed into the subjects cupped hands. This aliquot isrubbed over the entire surfaces of the hands for 20 6 5

46、s (frontand back) not reaching above the wrist.The hands are then heldmotionless away from the body and allowed to air dry forapproximately 30 6 5s.10.2.2 To continue the contamination of the hands, anadditional 1.5 mL aliquot of the test organism suspension isdispensed into the hands, distributed o

47、ver the hands for 20 65 s, and air dried for 30 6 5s.10.2.3 To complete the contamination, a final 1.5 mLaliquot of test organism suspension is dispensed into the hands,distributed over the hands for 20 6 5 s, and air dried for 90 65s(Table 1).NOTE 3The hands may still be wet after the 90 s.10.2.4 T

48、he total test organism suspension applied to thehands is 4.5 mL. Contamination may take approximately 5min. This method of contamination minimizes the loss of testorganism while spreading.10.3 Contamination ScheduleThe subjects hands arecontaminated with the test organism prior to the baselinebacter

49、ial sample collection and prior to each washing with thetest material. Table 2 illustrates a typical test. The number ofrepeated test washes may be reduced or eliminated at thediscretion of the investigator.10.4 Baseline RecoveryA baseline sample is taken aftercontamination to determine the number of marker organismssurviving on the hands. Bacterial sampling will follow theprocedures outlined in Section 12.11. Wash and Rinse Procedure11.1 Conduct the test in accordance with the use directionsfor the test material. If test ma