1、Designation: E 1173 01e1Standard Test Method forEvaluation of Preoperative, Precatheterization, orPreinjection Skin Preparations1This standard is issued under the fixed designation E 1173; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revi
2、sion, 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.e1NOTEThe title was editorially updated in March 2002.1. Scope1.1 The test method is designed to measure the re
3、duction ofthe resident microbial flora of 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
4、 standard does not purport to address all 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
5、 procedure requires a knowledge ofregulations pertaining to the protection of human subjects (1).22. Referenced Documents2.1 ASTM Standards:3E 1054 Practices for Evaluating Inactivators of Antimicro-bial Agents Used in Disinfectant, Sanitizer, Antiseptic, orPreserved ProductsE 1874 Test Method for E
6、valuation of Antibacterial Washesby Cup Scrub Technique3. Terminology3.1 active ingredienta substance 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 formu
7、lation withdemonstrated performance characteristics within a specificlaboratory.3.4 sampling fluida 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 a
8、fter treatment that prevents or inhibits the proliferationand/or survival of microorganisms.3.6 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
9、 chemical agents, called neutralizers,to curtail antimicrobial activity.4. Summary of Test Method4.1 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
10、days), exhibit acceptably highnormal flora counts on the skin sites to be used in testing (seeSection 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 applic
11、ation of a test formulation to skin sites, to countsobtained from those same sites prior to application 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)r
12、egion, and/or the median cubital region of the arm forprecatheterization preparations; and 3) 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 o
13、n both moist and dry skin sites. The1This test method is under the jurisdiction of ASTM Committee E35 onPesticides and is the direct responsibility of Subcommittee E35.15 on AntimicrobialAgents.Current edition approved April 10, 2001. Published July 2001. Originallypublished as E 1173 87. Last previ
14、ous edition E 1173 93.2The boldface numbers in parentheses refer to the list of references at the end ofthis standard3For 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
15、 the standards Document Summary page onthe ASTM website.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.sample times are 10 min, 30 min, and 6 h (or other appropriatetimes) post-treatment.4.2.2 Precatheterization PreparationMicrobial
16、 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 min, depending onthe test material being evaluated.4.2.3 Preinjection Pr
17、eparationA microbial sample is col-lected from the test site 30 sec post-treatment.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
18、product-testing, asdescribed in Practices E 1054, 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 paral
19、lel with thetest formulation.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
20、preparations, to maintain 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
21、 be used.6.3 SterilizerAny 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 th
22、e subject to recline.7. 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 c
23、ylinders, height ap-proximately 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 Teflont ScrubbersCan be fash-ioned in the labora
24、tory or purchased from most 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 skinsit
25、es.7.8 Sampling FluidDissolve 0.4 g KH2PO4, 10.1 gNa2HPO4, and 1.0 g isooctylphenoxypolyethoxyethanol5in 1L of 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 v
26、olumes and sterilize.7.9 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 E 1054).7.10 Plating MediumSoybean-casein digest agar (
27、3), withor without antimicrobial 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 Sit
28、es to be Used in Testing8.1 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
29、 atleast 3.0 log10/cm2greater 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 a
30、re the inguina, and the preferred 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
31、 of the template along theinguinal 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 p
32、ossible. In no instanceshould 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
33、from anterior to pos-terior, 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 for4A suitable covering is TELFAt non-adh
34、erent dressing, No. 3279, from theKendall Co.; Hospital Products; Boston, MA 02101.5Triton X-100, is available from Rohm and Haas Co., Philadelphia, PA.E117301e12baseline, and areas 2 through 4 for product exposure times of10 min, 30 min, and 6 $ h, respectively.NOTE 2Bacterial populations in the in
35、guina 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 the inguinalcrease laterally onto the (dry) surface of the upper thigh. Hence
36、, to providea fair test of a formulations antimicrobial efficacy, a “worst-case”sampling scheme is suggested. 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 sk
37、in-to-skin contact provides the moist environment conducive tobacterial growth. If preferred, however, the 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 substan
38、tially more subjects in order to demonstratestatistical significance of post-treatment reductions.8.1.2.2 Because 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 arera
39、ndomized one to each side.8.1.3 Abdominal treatment sites are to be located within5-by-5-in (12.7-by-12.7-cm) 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, pla
40、stic, cloth), the corners ofeach site are numbered 1, 2, 3, and 4 directly on the skin, usinga surgical skin 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 qua
41、drants of each site are used for thedifferent treatment exposure times of 10 min, 30 min, or $ 6h, and the 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 T
42、he skin sites selected for evaluation of the effective-ness of precatheterization skin preparations should 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 th
43、anthe detection limit of the sampling procedure and at least 1.0log10/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 preferred dry-skin areas are
44、 theclavicular region and the median cubital region of the arm.8.2.2 Test sites in the inguina are to be located 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 precatheterizati
45、on preparations are illustrated in Fig. 1Detail A. Sterile templates (for example, paper, plastic, cloth)are 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
46、 the treatment site, and a surgical skinFIG. 1 Illustration of Approximate Sampling Locations on Treatment Sites: Inguinal Crease, Abdomen, Clavicular Region, and MedianCubital Region of ArmE117301e13marker is used to demarcate the sampling areas. These arenumbered 1 through four at outside corners,
47、 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, depending on test product), 12 h, or
48、24 h, and theremaining sampling area is used for a baseline count. The testformulation and reference 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 s
49、kin 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, becausevariability in the bacterial counts may be reduced. A suitabledry-skin area is the median cubital region of the arm.8.3.2 The dry-skin site and sampling configuration used intesting preinjection preparations
