ASTM E2870-2013 Standard Test Method for Evaluating Relative Effectiveness of Antimicrobial Handwashing Formulations using the Palmar Surface and Mechanical Hand Sampling《使用掌面和机械手抽.pdf

1、Designation: E2870 13Standard Test Method forEvaluating Relative Effectiveness of AntimicrobialHandwashing Formulations using the Palmar Surface andMechanical Hand Sampling1This standard is issued under the fixed designation E2870; 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. Scope1.1 This test method covers and is designed to determine ther

3、elative effectiveness of antimicrobial handwashing agents inreducing transient microorganisms using a controlled hand-wash.1.2 Knowledge of microbiological techniques is requiredfor these procedures.1.3 This test method is used to evaluate topical antimicro-bial handwashing formulations.1.4 Performa

4、nce of this procedure requires the knowledgeof regulations pertaining to the protection of human subjects(21 CFR Parts 50 and 56).1.5 The values stated in SI units are to be regarded asstandard. No other units of measurement are included in thisstandard.1.6 This standard does not purport to address

5、all of thesafety concerns, if any, 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 8.1.2. Refere

6、nced Documents2.1 ASTM Standards:2E1054 Test Methods for Evaluation of Inactivators of Anti-microbial AgentsE1174 Test Method for Evaluation of the Effectiveness ofHealth Care Personnel Handwash FormulationsE2755 Test Method for Determining the Bacteria-Eliminating Effectiveness of Hand Sanitizer Fo

7、rmulationsUsing Hands of AdultsE2756 Terminology Relating to Antimicrobial and AntiviralAgentsE2784 Test Method for Evaluation of the Effectiveness ofHandwash Formulations Using the Paper Towel (Palmar)Method of Hand Contamination2.2 Other Standards:AATCC Test Method 147 Antibacterial Assessment of

8、Tex-tile Materials: Parallel Streak Method321 CFR Part 50 Protection of Human Subjects421 CFR Part 56 Institutional Review Boards43. Terminology3.1 DefinitionsFor definitions of terms used in thisdocument, see Terminology E2756.3.2 Definitions of Terms Specific to This Standard:3.2.1 active ingredie

9、nt, na substance added to a formula-tion specifically for the inhibition or inactivation of microor-ganisms.3.2.2 reference formulation, nformulation against whichthe activity of the test formulation is compared, for example, ahandwash without an active ingredient or a handwash with adifferent activ

10、e ingredient than the test formulation. Thisformulation is not considered a standard.3.2.3 test material, na product or formulation whichincorporates antimicrobial ingredients(s).3.2.4 test organism, nan applied inoculum of an organismthat has characteristics which allow it to be readily identified.

11、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.4. Summary of Test Method4.1 This test method uses adult subjects who have provideda written informed consent and whose hands h

12、ave been1This test method is under the jurisdiction of ASTM Committee E35 onPesticides, Antimicrobials, and Alternative Control Agents and is the directresponsibility of Subcommittee E35.15 on Antimicrobial Agents.Current edition approved April 1, 2013. Published May 2013. DOI: 10.1520/E2870-13.2For

13、 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 American Association of Textile Chemists and Colo

14、rists(AATCC), P.O. Box 12215, Research Triangle Park, NC 27709, http:/www.aatcc.org.4Available from U.S. Government Printing Office Superintendent of Documents,732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http:/www.access.gpo.gov.Copyright ASTM International, 100 Barr Harbor Drive,

15、PO Box C700, West Conshohocken, PA 19428-2959. United States1determined to be free from any apparent damage at the time ofparticipation in the study. Subjects are to refrain from use ofany antimicrobials for at least one week prior to the initiationof the test procedure (see 9.3).4.2 The test compar

16、es the activity of the two handwashformulations simultaneously on the same subject under stan-dardized and controlled conditions. Hands of the subjects areartificially contaminated with Escherichia coli. One hand ofthe subject is washed with the reference formulation and theother with the test formu

17、lation. The E. coli cells remaining onthe hands are recovered by the glove juice method of samplingusing a mechanical scrubber. In other methods, comparisonsbetween two products are made by testing two equivalentgroups of subjects. The objective of this test method is todetermine the relative differ

18、ence between two products testedon the same subjects and not to determine absolute reductionsin organism levels. By testing both products at the same timeon the same subjects with the same bacterial inoculum,variability is reduced.4.3 Effectiveness of the test material is determined after asingle wa

19、sh by comparing the numbers of viable test organismsrecovered after treatment with it and with the referenceformulation. As an example, a cleanser with an active ingre-dient (test formulation) can be compared to the cleanserwithout an active ingredient (reference formulation) to deter-mine the effec

20、t the active ingredient has on product perfor-mance.4.4 No baseline sampling of the hands is performed in thistest method. The inoculum volume to the palms is very small.Avolume of 100 L is applied to each palm and the palms andfingers are rubbed together. Spillage and loss do not occur, andorganism

21、s are evenly distributed across the palmar surfaceafter rubbing. As the objective of this test method is todetermine the relative difference between products and notabsolute reductions, baseline sampling is not performed.NOTE 1If an investigator wanted to compare the effect of washingwith a product

22、to not washing, this test could be conducted with one handserving as a baseline sample and the second hand treated with the testproduct.4.5 The investigator should be aware that there may behealth risks associated with the use of the test organism andprecautions similar to those referenced in 8.1 sh

23、ould beundertaken.5. Significance and Use5.1 Hand hygiene is important for preventing the spread ofmany types of infections.5.2 During routine activities, it is primarily the palmarsurface, comprising palms, fingers, and finger pads, of thehands that may become contaminated with transient microor-ga

24、nisms. The contamination could then be transferred toarticles touched or handled or to other parts of the body. Palmarcontamination is used in Test Method E2784.5.3 In Test Method E1174, incomplete drying of the experi-mentally contaminated hands dilutes the applied test product,thus compromising it

25、s activity. Application of a smaller vol-ume of the microbial test suspension keeps the soil load to areasonable level while allowing the hands to become visiblydry prior to application of the test material and referenceformulation. These modifications are aimed at producing abetter approximation of

26、 in-use conditions and a more realisticassessment of the test substance, thus providing a more reliableindication of product performance.5.4 UnlikeTest Methods E1174, E2755, and E2784, this testmethod enables a direct comparison between two formulationson the same subject. The test method also uses

27、a mechanicalscrubbing machine in conjunction with the glove juice tech-nique for more efficient recovery of viable test bacteria fromthe palms. The mechanical sampling results in greater recoveryof bacteria from the palms than conventional recoverymethods, such as massaging.6. Apparatus6.1 Colony Co

28、unterAny of several types may be used, forexample, Quebec Colony Counter.6.2 IncubatorAny incubator capable of maintaining thefollowing temperature: 35 6 2C.6.3 Shaker IncubatorAny incubator capable of maintain-ing the following temperature: 35 6 2C and capable ofshaking the culture medium at 120 to

29、 140 r/min.6.4 SterilizerAny suitable steam sterilizer capable of pro-ducing the conditions of sterilization.6.5 Timer (Stop-Clock)Type that can be read for minutesand seconds.6.6 Handwashing SinkA sink of sufficient size to permitsubjects to wash without touching hands to sink surface orother subje

30、cts.6.6.1 Water Faucet(s)To be located above the sink at aheight which permits the hands to be higher than the elbowduring the washing procedure. Faucet should maintain a flowrate of 3 L/min as determined in 10.4.6.6.2 Water Temperature Regulator and TemperatureMonitorTo set and maintain the water t

31、emperature at40 6 2C.6.7 Vortex MixerAny suitable vortex mixer capable ofmixing sample and diluent.6.8 Mechanical Scrubber5To mechanically sample thepalms for test bacteria (see Fig. 1). The machine contains twoartificial metallic paddles covered with an artificial turf for asmooth and nonslip surfa

32、ce. The drive mechanism is poweredby a 48V DC geared motor and has a variable speed from 50to 150 r/min produced by an electronic speed regulator. Thespecialized eccentric mechanism produces 12.7 cm long (totalstroke-length) horizontal reciprocating silent movement. Bothpaddles face upwards, paralle

33、l to the palms, and during the test,the complete system works to simulate the live activity ofmechanically sampling the hands.5The sole source of supply of the apparatus known to the committee at this timeis Trishul Equipment, Shiva Industrial Estate, Unit No. 107, First Floor, Lake Road,Bhandup Wes

34、t, Mumbai-400078, India. If you are aware of alternative suppliers,please provide this information to ASTM International Headquarters. Your com-ments will receive careful consideration at a meeting of the responsible technicalcommittee,1which you may attend.E2870 1326.9 SpectrophotometerAn instrumen

35、t that can measureoptical density at a wavelength of 620 nm.6.10 Adjustable or Fixed Volume Pipets and Sterile Tips1 mL capacity and 0.1 mL capacity.6.11 Sampling ContainersAny sterile or sterilizable con-tainer having a tight closure and sufficient capacity to hold75 mL of sampling solution (7.2).6

36、.12 CentrifugeFor the sedimentation of E. coli for con-centration.6.13 Sterile Centrifuge TubesMinimum of 15-mL capac-ity.6.14 Sterile ContainerAny sterile or sterilizable containerhaving the capacity to culture the amount of inoculum requiredfor testing.6.15 GlovesLoose-fitting, unlined, powder-fre

37、e gloveswhich possess no antimicrobial properties, or equivalent.Perform a zone of inhibition test, such asAATCC Test Method147, to evaluate the antibacterial activity. (Plastic bags (6.16)with low bioburden may be used in place of gloves.)6.16 Plastic BagsMay be used in place of gloves. Bagsshould

38、be approximately 30 18 cm, possess no antimicrobialproperties and have a low bioburden. Perform a zone ofinhibition test, such as AATCC Test Method 147, to evaluatethe antibacterial activity.6.17 Wrist Ties or TourniquetsAny item which will allowthe gloves (6.15) or plastic bags (6.16) to be secured

39、 to thesubjects wrists.6.18 Sterile Tissues or Paper TowelsAny sterile tissue orpaper towel that can be used to dry hands.7. Reagents and Materials7.1 Test SubstancesFollow the manufacturers directionsfor use of the test material and reference formulation. Ifdirections are not available, use the dir

40、ections provided in thistest method (10.5).7.2 Sampling SolutionDissolve 0.4 g monopotassiumphosphate (KH2PO4), 10.1 g disodium hydrogen phosphate(Na2HPO4), 1.0 g isooctylphenoxypolyethoxyethanol (forexample, Triton X-100), and appropriately validated neutraliz-ers in distilled water. Adjust pH to 7

41、.8 6 0.1 with 0.1 Nhydrochloric acid (HCl) or 0.1 N sodium hydroxide (NaOH)and bring volume to 1 L with distilled water. Sterilize in anautoclave and aseptically dispense 40-mL and 35-mL portionsinto sterile sampling containers (6.11).6NOTE 2Aneutralizer validation should be conducted according toTe

42、stMethod E1054 prior to the study. Test Method E1054 provides a list ofneutralizers appropriate for commonly used antimicrobial agents. In somecases, neutralization may be achieved by dilution alone.7.3 Dilution FluidSterile Butterfields buffered phosphatediluent7(or other suitable diluent) adjusted

43、 to pH 7.2 6 0.1 andcontaining an effective inactivator for the test material, ifnecessary.NOTE 3Inactivator is only required if neutralization of the testmaterial cannot be achieved upon dilution into the sampling solution (see7.2).7.4 Soybean-Casein Digest Agar with MUG(4methylumbelliferyl-b-D-glu

44、curonide)Sterile tryptic soyagar with MUG (0.5 g/L), used for the indication, recovery andgrowth of Escherichia species or other solid media appropri-ately validated to support the growth of the test organism. Withappropriate neutralizers, if required, according to Test MethodE1054.NOTE 4The MUG sub

45、strate is hydrolyzed by b-D-glucuronidase toyield a fluorescent end product, 4-methylumbelliferone. b-D-glucuronidase is possessed by E. coli (ATCC 10536).7.5 BrothSterile soybean-casein digest broth (tryptic soybroth) or other liquid media appropriate to support growth ofthe test organism.7.6 Soybe

46、an-Casein Digest AgarSterile tryptic soy agarfor growth of Escherichia species or other solid media appro-priately validated to support the growth of the test organism.7.7 Physiological SalineSterile. Used to prepare the finalinoculum.7.8 Ethanol or Isopropyl Alcohol Solution70 % ethanol orisopropyl

47、 alcohol in water (v/v) for hand decontamination.7.9 Chlorhexidine Skin CleanserAntiseptic skin cleansercontaining 4 % chlorhexidine gluconate (w/v) for hand decon-tamination.8. Test Organism8.1 Escherichia coli (ATCC 10536) is the test organism.The plating agar should be soybean casein digest agar

48、withMUG (7.4) or another suitable indicator. (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 an infection occurs, the antibiotic sensitivity profile

49、 shouldbe made available to the attending clinician. Following thesubjects contamination and wash with the formulation, adecontamination procedure should be performed (Section 11).8.2 Preparation of Test Organism Suspension:8.2.1 A homogeneous culture of E. coli (ATCC 10536) isused to inoculate the hands. The stock culture should be at least6Peterson, A. F., “The Microbiology of the Hands: Evaluating the Effects ofSurgical Scrubs,” Developments in Industrial Microbiology, Vol 14, 1973, pp.125130.7Butterfield, C. T., “The Selection