1、Designation: E1173 01 (Reapproved 2009)Standard Test Method forEvaluation of Preoperative, Precatheterization, orPreinjection Skin Preparations1This standard is issued under the fixed designation E1173; the number immediately following the designation indicates the year oforiginal adoption or, in th
2、e 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 The test method is designed to measure the reduction ofthe resident microbial flora of
3、 the skin.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 linear measure, in which case inches areused, and metric units follow in parentheses.1.4 This standard does not purport to address all
4、 of thesafety problems, 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.1.5 Performance of this procedure requires a knowledge ofregulat
5、ions pertaining to the protection of human subjects (1).22. Referenced Documents2.1 ASTM Standards:3E1054 Test Methods for Evaluation of Inactivators of An-timicrobial AgentsE1874 Test Method for Recovery of Microorganisms FromSkin using the Cup Scrub Technique3. Terminology3.1 active ingredienta su
6、bstance added to a formulationspecifically for the inhibition or inactivation of microorgan-isms.3.2 test formulationa formulation containing an activeingredient(s).3.3 internal reference formulationa formulation withdemonstrated performance characteristics within a specificlaboratory.3.4 sampling f
7、luida recovery fluid that may or may notcontain a neutralizer to inactivate the active ingredient(s) in testand internal reference formulations.3.5 persistenceprolonged or extended antimicrobial activ-ity after treatment that prevents or inhibits the proliferationand/or survival of microorganisms.3.
8、6 neutralizationa process that results in quenching theantimicrobial activity of a formulation. This may be achievedthrough dilution of the formulation to reduce the antimicrobialactivity, or through use of chemical agents, called neutralizers,to curtail antimicrobial activity.4. Summary of Test Met
9、hod4.1 These test methods are conducted on human subjectsselected randomly from a group of volunteers who, afterrefraining voluntarily from using topical and oral antimicrobi-als for at least two weeks (14 days), exhibit acceptably highnormal flora counts on the skin sites to be used in testing (see
10、Section 8).4.2 The antimicrobial activity of the preoperative, precath-eterization, or preinjection skin preparations is measured bycomparing microbial counts, obtained at various time intervalsafter application of a test formulation to skin sites, to countsobtained from those same sites prior to ap
11、plication of the testformulation. Skin sites recommended for use in testing are: 1)the inguinal region and the abdomen for preoperative skinpreparations; 2) the inguinal region, the subclavian (clavicular)region, and/or the median cubital region of the arm forprecatheterization preparations; and 3)
12、the median cubitalregion of the arm for preinjection skin preparations.4.2.1 Preoperative Skin PreparationMicrobial samplesare collected from the test sites a minimum of 3 times aftertreatment application on both moist and dry skin sites. Thesample times are 10 min, 30 min, and 6 h (or other appropr
13、iatetimes) post-treatment.4.2.2 Precatheterization PreparationMicrobial samplesare collected from the test sites a minimum of 3 times aftertreatment application on both moist and dry skin sites. Thesample times are “immediate,” 12 h, and 24 h post-treatment.The immediate sample may be 30 sec to 10 m
14、in, depending onthe test material being evaluated.4.2.3 Preinjection PreparationA microbial sample is col-lected from the test site 30 sec post-treatment.1This test method is under the jurisdiction of ASTM Committee E35 onPesticides and Alternative Control Agents and is the direct responsibility ofS
15、ubcommittee E35.15 on Antimicrobial Agents.Current edition approved Oct. 1, 2009. Published November 2009. Originallyapproved in 1987. Last previous edition approved in 2001 as E1173 011. DOI:10.1520/E1173-01R09.2The boldface numbers in parentheses refer to the list of references at the end ofthis s
16、tandard3For 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.1Copyright ASTM International, 100 Barr Harbor Drive,
17、PO Box C700, West Conshohocken, PA 19428-2959, United States.4.3 The fluid used for sampling the test sites must effec-tively quench (neutralize) the antimicrobial action of allformulations tested. The effectiveness of the inactivator mustbe demonstrated prior to initiation of product-testing, asdes
18、cribed in Practices E1054, and using in vivo techniquesconsistent with the cup-scrub technique (see Section 10).4.4 To ensure the internal validity of the test, an internalreference formulation having performance characteristicsknown to the laboratory should be tested in parallel with thetest formul
19、ation.5. Significance and Use5.1 These procedures should be used to test topicalantimicrobial-containing preparations that are intended to befast-acting and to reduce significantly the number of organismson intact skin immediately and, for preoperative and precath-eterization preparations, to mainta
20、in reductions for an extendedtime.6. Apparatus6.1 Colony CounterAny of several types may be used; forexample, Quebec colony counters and similar devices, orautomated, computerized plater/counter systems.6.2 IncubatorAny incubator that can maintain a tempera-ture of 30 6 2C may be used.6.3 Sterilizer
21、Any steam sterilizer that can produce theconditions of sterilization is acceptable.6.4 Timer (stopwatch)One that displays hours, minutes,and seconds.6.5 Examining TableAny elevated surface, such as a3-by-6-ft (0.9-by-1.8-meter) table with mattress or similarpadding to allow the subject to recline.7.
22、 Reagents and Materials7.1 Bacteriological Pipettes10.0 and 2.2-mL or 1.1-mLcapacity, available from most laboratory supply houses.7.2 Petri Dishes100 mm by 15 mm, for performingstandard plate counts, available from most laboratory supplyhouses.7.3 Scrubbing CupsAutoclavable cylinders, height ap-pro
23、ximately 1 in (2.5 cm), inside diameter of convenient size toplace on anatomical area to be sampled. Useful diametersrange from approximately 0.5 to 1.5 in (1.3 to 3.8 cm),depending on sites to be sampled.7.4 Rubber Policeman or Teflon ScrubbersCan be fash-ioned in the laboratory or purchased from m
24、ost laboratorysupply houses (whichever type is selected, it should be usedthroughout the course of testing).7.5 Testing Formulation, including directions for use.7.6 Sterile Gauge PadsUsed to cover treated skin sites.7.7 Sterile Drape or Dressing4Used to cover treated skinsites.7.8 Sampling FluidDis
25、solve 0.4 g KH2PO4, 10.1 gNa2HPO4, and 1.0 g isooctylphenoxypolyethoxyethanol in 1 Lof distilled water. Inactivator(s) specific for the antimicrobialactive(s) in the test and reference formulations may be in-cluded, if appropriate. Adjust to pH 7.8. Dispense in appropri-ate volumes and sterilize.7.9
26、 Dilution FluidButterfields (2) phosphate-bufferedwater adjusted to pH 7.2, or other suitable diluent, which mayor may not contain antimicrobial inactivators specific for thetest and reference formulations (see Practices E1054).7.10 Plating MediumSoybean-casein digest agar (3), withor without antimi
27、crobial inactivators.7.11 Sterile Template MaterialUsed to demarcate the skinsites; made from paper, plastic, or cloth, for example.7.12 Surgical Skin MarkerUsed to mark the skin sites.NOTE 1Some markers contain crystal violet, which is inhibitory tomany bacteria.8. Skin Sites to be Used in Testing8
28、.1 Preoperative Skin Preparations8.1.1 The skin sites selected for evaluation of the effective-ness of preoperative skin preparations should include bodyareas that may be surgical sites and should include both moistand dry skin areas. Bacterial baseline populations should be atleast 3.0 log10/cm2gre
29、ater on moist skin sites than thedetection limit of the sampling procedure and at least 2.0log10/cm2greater than the detection limit on dry skin sites.High baseline counts are desired, because variability in thebacterial counts may be reduced. The preferred moist-skinareas are the inguina, and the p
30、referred dry-skin area is thelower abdomen below the umbilicus.8.1.2 Using a 1.5-by-5-in (3.8-by-12.7-cm) sterile template(for example, paper, plastic, cloth), treatment sites are delin-eated on the uppermost inner aspects of both thighs (theinguina), centering the long axis of the template along th
31、einguinal crease immediately below the groin and marking thecorners, using a surgical skin marker. If, due to a subjectsanatomy, the treatment site cannot be centered along theinguinal crease, the site should be positioned on the upper,inner thigh as close to the crease as possible. In no instancesh
32、ould sampling be performed on areas not having skin-to-skincontact. The site is then divided on the long axis into1-by-1.5-in (2.5-by-3.8-cm) sampling areas, allowing forspaces of about 0.25 in (about 0.6 cm) between each of the fourareas.8.1.2.1 Sampling areas are numbered from anterior to pos-teri
33、or, beginning with 1 and proceeding perineally to 4. To testa “worst-case scenario” for efficacy of preoperative skinpreparations (see Note 2, below), the areas are not randomizedfor baseline and exposure times. Area 1 is always used forbaseline, and areas 2 through 4 for product exposure times of10
34、 min, 30 min, and 6 $ h, respectively.NOTE 2Bacterial populations in the inguina are known to be hetero-geneous, with counts tending to increase proceeding from the upperreaches of the inguinal crease perineally toward convergence of theinguina at the gluteal fold, and to decrease proceeding from th
35、e inguinalcrease laterally onto the (dry) surface of the upper thigh. Hence, to providea fair test of a formulations antimicrobial efficacy, a “worst-case”4The sole source of supply of the apparatus (TELFA non-adherent dressing, No.3279) known to the committee at this time is Kendall Co.; Hospital P
36、roducts;Boston, MA 02101. If you are aware of alternative suppliers, please provide thisinformation to ASTM International Headquarters. Your comments will receivecareful consideration at a meeting of the responsible technical committee,1whichyou may attend.E1173 01 (2009)2sampling scheme is suggeste
37、d. Baseline sampling is performed in sam-pling area 1 (lowest count area), with post-treatment sampling progres-sively deeper, and sampling areas must be confined to the inguinal creasewhere skin-to-skin contact provides the moist environment conducive tobacterial growth. If preferred, however, the
38、sampling areas may berandomized to baseline and post-treatment sampling times. Because of theincreased variance of the count data, it is likely that such a design willrequire testing of substantially more subjects in order to demonstratestatistical significance of post-treatment reductions.8.1.2.2 B
39、ecause of constraints imposed by the anatomicalarea, sampling cylinders used for the inguinal sites must be #1in(# 2.54 cm) in diameter.8.1.2.3 The test formulation and reference material arerandomized one to each side.8.1.3 Abdominal treatment sites are to be located within5-by-5-in (12.7-by-12.7-c
40、m) sites below and to the right or leftof the umbilicus, approximately midway between the umbili-cus and the pubis. Using a 5-by-5-in (12.7-by-12.7-cm) steriletemplate (for example., paper, plastic, cloth), the corners ofeach site are numbered 1, 2, 3, and 4 directly on the skin, usinga surgical ski
41、n marker. Numbering is to be the same for allabdominal sites: number 1 is placed at the top corner to thesubjects right, and numbers 2, 3, and 4 are assigned in orderclockwise from 1. Three quadrants of each site are used for thedifferent treatment exposure times of 10 min, 30 min, or $ 6h, and the
42、remaining quadrant is used for a baseline count. Thetest formulation or reference material should be randomized tothe four quadrants of each site.8.2 Precatheterization Skin Preparations8.2.1 The skin sites selected for evaluation of the effective-ness of precatheterization skin preparations should
43、includebody areas that may be catheterization sites and should includeboth moist and dry skin areas. Bacterial baseline populationsshould be at least 3.0 log10/cm2greater on moist skin sites thanthe detection limit of the sampling procedure and at least 1.0log10/cm2greater than the detection limit o
44、n dry skin sites.High baseline counts are desired, because variability in thebacterial counts may be reduced. The preferred moist-skinareas are the inguina, and the preferred dry-skin areas are theclavicular region and the median cubital region of the arm.8.2.2 Test sites in the inguina are to be lo
45、cated and evaluatedas specified for testing of preoperative skin preparations (see8.1.2, Note 1, and Fig. 1).8.2.3 The dry skin sites and sampling configurations used intesting precatheterization preparations are illustrated in Fig. 1Detail A. Sterile templates (for example, paper, plastic, cloth)ar
46、e fashioned for the sampling configuration such that theyaccommodate the diameter of the sampling cylinder, plus atleast 0.5 in (1.25 cm) between the 4 sampling areas. Thetemplate is applied to the treatment site, and a surgical skinmarker is used to demarcate the sampling areas. These arenumbered 1
47、 through four at outside corners, beginning at thesubjects upper right and proceeding clockwise in the clavicu-lar region, and beginning proximally and proceeding distallyon the arm. Three sampling areas of the site are used fordifferent treatment exposure times of “immediate” (30 sec to10 min, depe
48、nding on test product), 12 h, or 24 h, and theFIG. 1 Illustration of Approximate Sampling Locations on Treatment Sites: Inguinal Crease, Abdomen, Clavicular Region, and MedianCubital Region of ArmE1173 01 (2009)3remaining sampling area is used for a baseline count. The testformulation and reference
49、material should be randomized tothe treatment sites, right or left, and exposure times andbaseline should be randomized to the four quadrants of eachsite.8.3 Preinjection Skin Preparations8.3.1 The skin site selected for use in evaluating theeffectiveness of preinjection skin preparations should representa body area that is commonly used for transepidermal injectionor phlebotomy. Bacterial baseline populations should be atleast 1.0 log10/cm2greater than the detection limit of thesampling procedure. High baseline counts are desired, becausevariabi
