ASTM E1174-2006 Standard Test Method for Evaluation of the Effectiveness of Health Care Personnel Handwash Formulations《卫生保健人员洗手模式效果的评估用标准试验方法》.pdf

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1、Designation: E 1174 06Standard Test Method forEvaluation of the Effectiveness of Health Care PersonnelHandwash Formulations1This standard is issued under the fixed designation E 1174; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision,

2、 the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) indicates an editorial change since the last revision or reapproval.1. Scope1.1 This test method covers the determination of the effec-tiveness of antimicrobial handwashing agents for

3、the reductionof transient microbial flora when used in a handwashingprocedure.1.2 A knowledge of microbiological techniques is requiredfor these procedures.1.3 In this test method, metric units are used for allapplications, except for distance in which case inches are usedand metric units follow in

4、parentheses.1.4 This test method may be used to evaluate topicalantimicrobial handwash formulations.1.5 Performance of this procedure requires the knowledgeof regulations pertaining to the protection of human subjects.21.6 This standard does not purport to address all of thesafety concerns, if any,

5、associated 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:3E

6、1054 Test Methods for Evaluation of Inactivators ofAntimicrobial Agents3. Terminology3.1 Definitions:3.1.1 active ingredienta substance added to a formulationspecifically for the inhibition or inactivation of microorgan-isms.3.1.2 cleansing washa non-antimicrobial wash intendedto remove gross soil o

7、r 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 healthcare personnel handwasha cleanser or water-less agent intended to reduce transient bacteria on the hands.3.1.4 neutralizationa pro

8、cess which results in quenchingthe antimicrobial activity of a test material. This may beachieved through dilution of the test material(s) to reduce theantimicrobial activity, or through the use of chemical agents,called neutralizers, to eliminate antibacterial activity.3.1.5 resident microorganisms

9、microorganisms that liveand multiply on the skin, forming a permanent population.3.1.6 test formulationa formulation which incorporatesantimicrobial ingredient(s).3.1.7 test organisman applied inoculum of an organismthat has characteristics which allow it to be readily identified.The test organism i

10、s used to simulate a transient topicalmicrobial contaminant. It may also be referred to as a markerorganism, bacterial simulant, or bacterial contaminant.3.1.8 transient microorganismsorganisms from the envi-ronment that contaminate but do not normally colonize theskin.4. Summary of Test Method4.1 T

11、his test method is conducted on a group of volunteerpanelists who have refrained from using topical antimicrobialformulations for at least one week prior to the initiation of thetest. Activity of the test material is measured by comparing thenumber of test organisms recovered from artificially conta

12、mi-nated hands after use of a handwashing formulation to thenumber recovered from contaminated hands not exposed to thetest formulation. The method describes specific procedures tobe followed using Serratia marcescens as the test organism.The activity of the test material may be measured following a

13、single wash and multiple washes in a single day using aneutralization recovery method.4.2 An alternative test organism is Escherichia coli. Culturemedia and incubation conditions appropriate for this organismshould be employed. The investigator should also be aware thatthere may be health risks asso

14、ciated with the use of this1This test method is under the jurisdiction of ASTM Committee E35 onPesticides and Alternative Control Agents and is the direct responsibility ofSubcommittee E35.15 Antimicrobial Agents.Current edition approved March 1, 2006. Published April 2006. Originallyapproved in 198

15、7. Last previous edition approved in 2000 as E 1174 00.2Federal 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 standard

16、s Document Summary page onthe ASTM website.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.organism and precautions similar to those referenced in 8.2should be undertaken.5. Significance and Use5.1 The procedure may be used to test t

17、he effectiveness ofantimicrobial handwashing agents. The test formulations maybe designed for frequent use to reduce the transient bacterialflora on hands.6. Apparatus6.1 Colony CounterAny of several types may be used, forexample, Quebec Colony Counter.6.2 IncubatorAny incubator capable of maintaini

18、ng 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 ofproducing the conditions of sterilization is acceptable.6.4 Timer (Stop-clock)One that can be re

19、ad 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 during the washing procedur

20、e.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.47.2 Water Dilution BottlesAny sterilizable glass containerhaving a 150 to 200 mL capacity and tight

21、closures may beused.57.3 Erlenmeyer Flask2-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 solu

22、tion of potassium hydroxidein 15 parts water 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

23、 then brought up to 100 parts byaddition of 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 th

24、is test method.7.6 GlovesLoose-fitting, unlined, powder-free gloveswhich possess no antimicrobial properties, or equivalent.6(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 isooctylphenoxypolyethoxyethanol7andwith a

25、ppropriately validated neutralizers in 1 L 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.87.8 Dilution FluidSterile Butterfields Buffer9or othersuitable diluent, adjusted to pH 7.2 with effective neutralize

26、rfor the test material.Adjust pH with 0.1 N HCl or 0.1 N NaOH.See Test Methods E 1054.7.9 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 liquidmed

27、ia appropriate to support growth of the test 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 sh

28、ould includea suitable indicator (for example, MUG10). (WarningTheapplication of microorganisms to the skin may involve a healthrisk. Prior to applying the test organism to the skin, theantibiotic sensitivity profile of the strain should be determined.If the strain is not susceptible to gentamicin,

29、do not use it. If aninfection occurs, the antibiotic sensitivity profile should bemade available to the attending clinician. Following the sub-jects last contamination and wash with the formulation, the4Presterilized/disposable bacteriological pipettes are available from most locallaboratory supply

30、houses.5Milk dilution bottles of 160-mL capacity having a screw-cap closure areavailable from most local laboratory supply houses.6The sole source of supply of the apparatus (Pharmaseal 8873C, (sterile) FlexamLatex Procedure Glove) known to the committee at this time is AmericanPharmaseal Laboratori

31、es, Glendale, CA 91209. If you are aware of alternativesuppliers, please provide this information to ASTM International Headquarters.Your comments will receive careful consideration at a meeting of the responsibletechnical committee,1which you may attend. A zone of inhibition test such asAATCC Test

32、Method 90-1965 may be used to evaluate antimicrobial properties ofgloves, AATCC Test Methods, American Association of Textile Chemists andColorist, 1968 Technical Manual, Section B-75.7The sole source of supply of the apparatus known to the committee at this timeis Triton X-100, Rohm and Haas Co., P

33、hiladelphia, PA. If you are aware ofalternative suppliers, please provide this information to ASTM InternationalHeadquarters. Your comments will receive careful consideration at a meeting of theresponsible technical committee,1which you may attend.8Peterson, A. F., “The Microbiology of the Hands: Ev

34、aluating the Effects of theSurgical Scrubs,” Developments in Industrial Microbiology, Vol 14, 1973, pp.125130.9Horowitz, W., (Ed.), Offcial Methods of Analysis of the AOAC, 17th Ed., Sec.6.3.03 A.(f), Chapter 6, 2000, p. 10. Official Methods of Analysis of AOACInternational, Gaitherburg, MD.10United

35、 States Pharmacopeia 28: United States Pharmacopeial Convention,Inc., Rockville, MD, Chapter entitled “Microbial Limits Test.” The MUG (4-methylumbelliferyl-b-D-gluconride) substrate is hydrolyzed by b-D-gluconridase toyield a fluorescent end product, 4-methylumbelliferone. b-D-gluconridase is pos-s

36、essed by E. coli (ATCC 11229). MUG is incorporated into the appropriate growthmedium at 0.05 g/L.E1174062subjects hands are to be sanitized by scrubbing with 70%isopropanol solution or equivalent. The purpose of this alcoholscrub is to destroy residual test organisms on the skin.)8.3 Preparation of

37、Test Organism Suspension8.3.1 S. marcescensA homogeneous culture is used toinoculate the hands. The stock culture, frozen or lypholized,should be at least two 24-h soybean-casein digest broth (7.10)transfers from the original ATCC culture, but there should beno more than four transfers removed from

38、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 stock culture of S. marcescens/100mL of broth to yield the volume necessary to complete thestudy. Incubate for 24 6 4hat256 2C. Broth shoul

39、d 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 original ATCC culture, but no morethan five transfers removed from the ATCC culture. From thestock culture of Escherichia coli (ATCC 11229

40、), 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 study. Incubate for 24 6 4hat3562C.8.4 Swirl or shake suspension before the withdrawal of eachaliquot. Assay the suspension for number of

41、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 period.9. Subjects9.1 Recruit a sufficient number of healthy adult humanvolunteers who have no clinical evidence of dermatosis, openwounds,

42、 hangnails, or other skin disorders.9.2 Instruct subjects to avoid contact with antimicrobialproducts (other than the test material as dispensed for each testwash) for the duration of the test and for at least one week priorto the test. This restriction includes antimicrobial-containingantiperspiran

43、ts, deodorants, shampoos, lotions, and soaps, alsosuch materials as acids, bases, and solvents. Bathing in biocidetreated pools, hot tubs, or spas should be avoided. Subjects areto be provided with a kit of nonantimicrobial personal careproducts for exclusive use during the test and rubber gloves to

44、be worn when contact with antimicrobial or harsh chemicalscannot be avoided.10. Procedure10.1 After subjects have refrained from using antimicrobialformulations for at least 7 days, they perform a 30 s cleansingwash (7.4) in the same manner that is described for the test andcontrol formulations. Thi

45、s procedure removes oil and dirt andfamiliarizes the panelists with the washing technique. For thisand all other washes and rinses, the water temperature isadjusted to 40 6 2C and the water flow rate to 4 L per minute.This may be accomplished by placing a 2000 mL glass beakeror flask under each spig

46、ot 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 s.10.2 Hand ContaminationA liquid suspension of the testorganism containing between 5.0 3 108and 1.0 3 109cfu/mLis used. See Table 1.10.

47、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 s (frontand back) not reaching above the wrist. The hands are then heldmotionless away from the body and allowed to air dry forappro

48、ximately 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 over 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 s

49、uspension is dispensed into the hands,distributed over the hands for 20 6 5 s, and air dried for 90 65s(Table 1).NOTE 1The hands may still be wet after the 90 s.10.2.4 The 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 baselinebacterial sample collection and prior to each washing with thetest material. Tab