ASTM E2149-2001 Standard Test Method for Determining the Antimicrobial Activity of Immobilized Antimicrobial Agents Under Dynamic Contact Conditions 《在动态接触条件下固定抗菌剂抗菌活性测定的标准试验方法》.pdf
《ASTM E2149-2001 Standard Test Method for Determining the Antimicrobial Activity of Immobilized Antimicrobial Agents Under Dynamic Contact Conditions 《在动态接触条件下固定抗菌剂抗菌活性测定的标准试验方法》.pdf》由会员分享,可在线阅读,更多相关《ASTM E2149-2001 Standard Test Method for Determining the Antimicrobial Activity of Immobilized Antimicrobial Agents Under Dynamic Contact Conditions 《在动态接触条件下固定抗菌剂抗菌活性测定的标准试验方法》.pdf(4页珍藏版)》请在麦多课文档分享上搜索。
1、Designation: E 2149 01Standard Test Method forDetermining the Antimicrobial Activity of ImmobilizedAntimicrobial Agents Under Dynamic Contact Conditions1This standard is issued under the fixed designation E 2149; the number immediately following the designation indicates the year oforiginal adoption
2、 or, in the case of revision, 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 is designed to evaluate the resistance ofnon-leaching anti
3、microbial treated specimens to the growth ofmicrobes under dynamic contact conditions. This dynamicshake flask test was developed for routine quality control andscreening tests in order to overcome difficulties in usingclassical antimicrobial test methods to evaluate substrate-bound antimicrobials.
4、These difficulties include ensuring con-tact of inoculum to treated surface (as in AATCC 100),flexibility of retrieval at different contact times, use of inap-propriately applied static conditions (as in AATCC 147),sensitivity, and reproducibility. This test also allows for theversatility of testing
5、 contamination due to such things as hardwater, proteins, blood, serum, various chemicals, and othercontaminates or physical/chemical stresses or manipulations ofthe specimens of interest.1.2 Surface antimicrobial activity is determined by compar-ing results from the test sample to simultaneously ru
6、n controls.1.3 The presence of a leaching antimicrobial is both pre-and post-determined by the presence of a zone of inhibition.1.4 This test method should be performed only by thosetrained in microbiological techniques.1.5 This standard may involve hazardous materials, opera-tions, and equipment. T
7、his standard does not purport toaddress all of the safety concerns, if any, associated with itsuse. It is the responsibility of the user of this standard toestablish appropriate safety and health practices and deter-mine the applicability of regulatory limitations prior to use.2. Referenced Document
8、s2.1 ASTM Standards:E 1054 Practices for Evaluating Inactivators of Antimicro-bial Agents Used in Disinfectant, Sanitizer, Antiseptic, orPreserved Products2.2 Other Documents:AATCC Test Method 147-1998 Antibacterial Activity As-sessment of Textile Materials: Parallel Streak Method.American Associati
9、on of Textile Chemists and Colorists,RTP, NCAATCC Test Method 100-1999 Antibacterial Finishes onFabrics, Evaluation of American Association of TextileChemists and Colorists, RTP, NC3. Summary of Test Method3.1 Immobilized antimicrobial agents, such as surfacebonded materials, are not free to diffuse
10、 into their environmentunder normal conditions of use. Test methods such as AATCC147 that are directly dependent on the ready leachability of theantimicrobial agent from the treated fabric are inappropriate forevaluating immobilized antimicrobial agents. The followingtest method ensures good contact
11、 between the bacteria and thetreated fiber, fabric, or other substrate by constant agitation ofthe test specimen in a bacterial suspension during the testperiod. The test is suitable for evaluating stressed or modifiedspecimens when accompanied by adequate controls.NOTE 1Stresses may include laundry
12、, wear and abrasion, radiationand steam sterilization, UV exposure, solvent manipulation, temperaturesusceptibility, or similar physical or chemical manipulation.4. Significance and Use4.1 The antimicrobial activity of a substrate-bound antimi-crobial is dependent upon direct contact of microbes wit
13、h theactive chemical agent. This test determines the antimicrobialactivity of treated specimen by shaking samples of surfacebound materials in a concentrated bacterial suspension for aone hour contact time or other contact times as specified by theinvestigator. The suspension is serially diluted bot
14、h . beforeand after contact and cultured. The number of viable organismsin the suspension is determined and the percent reduction iscalculated based on initial counts or on retrievals from appro-priate untreated controls.NOTE 2This method is intended for those surfaces having a percentreduction acti
15、vity of 50 % to 100 % for the specified contact time.5. Apparatus5.1 Sterilizer,5.2 Incubator,1This 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 June 10, 2001. Publishe
16、d August 2001.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.5.3 Spectrophotometer,5.4 Shaker, Wrist ActionA Wrist Action Shaker is recom-mended but other means of agitation such as reciprocal actionshakers may be satisfactory for r
17、outine testing. Shaker mustensure good agitation. Rotary shakers are unacceptable.5.5 Water Bath,5.6 Vortex Mixer,5.7 Glassware,5.7.1 Contact Flask, 250 ml Erlenmeyer flask, capped,autoclavable.5.7.2 Dilution Vessels,5.7.3 Pipettes,5.8 Agar, bore 8-mm diameter.6. Reagents6.1 Buffer SolutionFor test
18、specimen which might alterthe pH of the system, Sorensens Phosphate Buffer (pH 6.8) isrecommended.2Other appropriate buffers must be shown not tocause a reduction or increase in bacterial numbers by priortesting at the intended use concentration. For all other samples,the following solution is recom
19、mended and is prepared fromreagent grade chemicals. For buffer stock solution (0.25MKH2PO4): Prepare a fresh solution at least once every 6 monthsas follows: Weigh 34 6 0.1 g of potassium dihydrogenphosphate into a 100 ml beaker. Add 500 ml of distilled water.Adjust pH to 7.2 6 0.1 with a dilute sol
20、ution of NaOH. Diluteto 1000 ml; transfer to a flask and store at 4C. For workingbuffer solution (0.3mM KH2PO4): Prepare a fresh solution atleast once every 2 months as follows: Transfer 1 ml 6 0.01 mlof stock buffer solution, with a sterile pipette to flask contain-ing 800 ml of distilled water. Ca
21、p and sterilize.6.2 Media:6.2.1 Nutrient Broth (Difco Laboratories, Detroit, MI orequivalent) or media appropriate of organism selected.6.2.2 Tryptone Glucose Extract Agar (Difco Laboratories,Detroit, MI or equivalent) or media appropriate for organismselected.6.3 Wetting Agent SurfactantAgents must
22、 be shown not tocause a reduction or increase in bacterial numbers by priortesting at the intended use concentration.NOTE 3Dow Corning, Midland, MI Q2-5211 at 0.01 % final dilutionor equivalent can be used.7. Test Organism7.1 Klebsiella pneumoniae, American Type Culture Collec-tion No. 4352. Other o
23、rganisms may be used at the discretionof the investigator.7.1.1 Cultures of the test organism should be maintainedaccording to good microbiological practice and checked forpurity, on a routine basis. Consistent and accurate testingrequires maintenance of a pure, uncontaminated test culture.Avoid con
24、tamination by use of good sterile technique in platingand transferring. Avoid mutation or reversion by strict adher-ence to monthly stock transfers. Check culture purity bymaking streak plates periodically and observing for a singlespecies characteristic type of colonies.NOTE 4A glossiness in the br
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