1、Designation: E 2406 04Standard Test Method forEvaluation of Laundry Sanitizers and Disinfectants for Usein High Efficiency Washing Operations1This standard is issued under the fixed designation E 2406; the number immediately following the designation indicates the year oforiginal adoption or, in the
2、 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 sanitizing/disinfectant laundry detergents/ad
3、ditives for use in high effi-ciency (HE) automatic clothes washing operations that typi-cally utilize very low wash water volumes. This test method isdesigned to provide testing with representative vegetativebacteria but can also be designed to accommodate the testing offungi and viruses.NOTE 1Stand
4、ard test method Test Method E 2274 is the recom-mended method to evaluate sanitizing/disinfectant laundry detergent/additives for use in traditional high wash water volume automatic clotheswashing operations.1.2 Knowledge of microbiological techniques is requiredfor these procedures.1.3 In this meth
5、od metric units are used for all applications,except for distance in which case inches are used.1.4 This standard does not purport to address 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 p
6、ractices and determine the applica-bility of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D 1193 Specification for Reagent WaterE 1054 Practices for Evaluating Inactivators of Antimicro-bial Agents Used in Disinfectant, Sanitizer, Antiseptic, orPreserved ProductsE 2
7、274 Test Method for Evaluation of Laundry Sanitizersand Disinfectants2.2 AATCC Standard:Test Method 70-1997 Water Repellency; Tumble Jar Dy-namic Absorption Test32.3 AOAC Standard:Official Methods of Analysis of AOAC International Chap-ter 6: Disinfectants, 17th ed., 200042.4 EPA Standard:DIS/TSS 13
8、 Laundry AdditivesDisinfection and Sanitiza-tion, U.S. Environmental Protection Agency, Office ofPesticide Programs, April 198052.5 Federal Standard:40 CFR, Part 160 Good Laboratory Practice Standards62.6 Canadian Standard:T-1-215 Canadian Pest Management Regulatory StandardsTrade Memorandum Oct, 19
9、80.73. Terminology3.1 Definitions:3.1.1 active antimicrobial ingredienta substance added toa formulation intended specifically for the inhibition or inac-tivation of microorganisms.3.1.2 antimicrobial agent(s)an active ingredient designedto suppress the growth or action of microorganisms.3.1.3 carri
10、er count controlprocedure used to determinethe initial number of microorganisms on a fabric carrierfollowing the inoculation and drying procedure.3.1.4 diluentsterile deionized water, sterile distilled wateror sterile synthetic AOAC hard water that may be used toprepare the active test formulation,
11、vehicle control or productcontrol for use in the test procedure.3.1.5 diluted product solutiontest formulation, vehiclecontrol, or product control diluted to use concentration.1This test method is under the jurisdiction of ASTM Committee E35 onPesticides and is the direct responsibility of Subcommit
12、tee E35.15 on AntibacterialAgents.Current edition approved Oct. 1, 2004. Published November 2004.2For 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 Docum
13、ent Summary page onthe ASTM website.3Available from American Association of Textile Chemists and Colorists(AATCC), One Davis Dr., P.O. Box 12215, Research Triangle Park, NC 27709-2215.4Available from AOAC International, Washington, DC.5Available from United States Environmental Protection Associatio
14、n (EPA),Ariel Rios Bldg., 1200 Pennsylvania Ave., NW, Washington, DC 20460.6Available from U.S. Government Printing Office Superintendent of Documents,732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401.7Available from Canadian Standards Association (CSA), 178 Rexdale Blvd.,Toronto, ON Cana
15、da M9W1R3.1Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.3.1.6 neutralizationa process that results in quenching theantimicrobial activity of a test formulation. This may beachieved by dilution of the test formulation(s) to reduce t
16、heconcentration of the antimicrobials, or through the use ofchemical agents, called neutralizers, to suppress antibacterialactivity.3.1.7 numbers controlin assessing sanitizer level perfor-mance, procedure used to determine the number of microor-ganisms remaining on the fabric carriers and in the wa
17、sh waterfollowing the test procedure in the presence of the diluent. Thismay also be performed using diluent or phosphate bufferdilution water with surfactant.3.1.8 product controla formulation with or without anactive ingredient(s) used for comparison to the test formula-tion.3.1.9 test formulation
18、a formulation containing an antimi-crobial agent(s).3.1.10 vehicle controlthe test formulation without theactive ingredient(s) used for comparison to the test formula-tion.3.1.11 wash waterthe liquid contained in the exposurechamber previously exposed to either uninoculated fabric orfabric inoculate
19、d with the challenge microorganism.4. Summary of Test Method4.1 Under simulated laundry conditions, sets of inoculatedfabric swatches are placed into low volumes of diluted productsolution and agitated. After a specified contact time, the washwater and the test fabric are individually cultured eithe
20、rquantitatively (sanitizer efficacy) or qualitatively (disinfectantefficacy).NOTE 2See appropriate regulatory guidance document for the mini-mum number of replicates required to make a specific claim.5. Significance and Use5.1 The procedure in this test method is used to evaluate theeffectiveness of
21、 a test reagent (antimicrobial agent/activeingredient) or formulation to reduce or completely kill bacterialpopulations on contaminated fabrics and in wash water follow-ing a single wash under simulated low wash volume condi-tions. (See Table 1.)6. Apparatus6.1 Colony CounterAny of several types may
22、 be used, forexample, Quebec.6.2 IncubatorAny incubator that can maintain the opti-mum temperature 6 2C for growth of the challenge microor-ganism(s).6.3 SterilizerAny suitable steam sterilizer producing theconditions of sterility.6.4 Timer (Stop-clock)Any device that can be read forminutes and seco
23、nds.6.5 Exposure ChamberContainer with closure that canwithstand sterilization. Dimension and volume capacity shouldbe consistent for use in Test Method E 2274.NOTE 3Standard lids may form a vacuum seal when steam sterilized.To avoid, prior to sterilization place a piece of paper between lid and jar
24、.6.6 Stainless Steel SpindlesSpindles are fabricated from asingle continuous piece of stainless steel wire (116 in. diameterand bent to contain 3 horizontal extensions, 2 in. longconnected by 2 vertical sections approximately 2 in. long).They are shaped so that vertical sections form 150 anglewhere
25、the free ends of the 2 outer horizontal extensions aresharpened to a point. This will be used as scaffolding for initialwrapping of fabric ballast.6.7 AgitatorTumbling device intended to rotate ExposureChamber through 360 vertical orbit of 4 to 8 in. diameter at 45to 60 rpm or a comparable tumbling
26、devices such as Launder-ometer or Tumble Jar described in AATCC 70-1997.6.8 Micropipettor (and Pipet Tips), suitable to deliver 0.01to 0.03 mL volume.6.9 Forceps, large and small, sterile.6.10 Safety Pins, sterile.6.11 Stapler and Staples.6.12 Balance, with a platform to accommodate 15 6 0.1 gof tes
27、t fabric.6.13 Sterile Glass Beads,3to4mm.6.14 Filter Sterilization System for Media and ReagentsAmembrane or cartridge filtration system (0.22 m pore diam-eter). Required for sterilizing heat-sensitive solutions.6.15 Membrane Filtration System for Capture of the TestOrganism(s)Sterile 47 mm diameter
28、 membrane filters (0.45m pore diameter) and holders for such filters.7. Reagents and Materials7.1 Petri Dishes, sterile 100 by 15 mm. Required forperforming standard plate counts and used in preparation ofcontaminated fabric carriers.7.2 Bacteriological Pipets, sterile, various sizes.TABLE 1 Typical
29、 Use PatternsUsage PatternFabric:Wash WaterRatioWash WaterVolumeSpindle WireRequirementDeep Fill Top Load Washers $ 1:5-15 75-225 mL RetainHigh Efficiency/Horizontal FrontLoad Washers 1:5 75 mL OmitFIG. 1 Stainless Steel Spindle Schematic(Top View and Side View)E24060427.3 Test Fabric, approximately
30、 80 by 80 threads/in.bleached, desized, plain-weave cotton print cloth and withoutbluing or optical brighteners.8NOTE 4Other test fabrics/blends may be used at the discretion of theinvestigator.7.4 Dilution Fluid, AOAC Phosphate buffer dilution wateror other suitable diluent containing appropriate n
31、eutralizers forserial dilution of test samples.7.5 Water for Dilution of Formulations Under Test:7.5.1 Water, sterile, deionized or distilled, equivalent to orbetter than Type 3, see Specification D 1193.7.5.2 AOAC Synthetic Hard Water.7.5.3 All water used for preparation of test solutions shall bes
32、terile.7.6 Purity of ReagentsReagent grade chemicals shall beused in all tests.7.6.1 Sodium carbonate.7.6.2 Alkaline nonionic wetting agent with HLB(hydrophilic-lipophilic balance) value of approximately 13.Prepare solution containing 0.5 % nonoxynol-10 class ofethoxylated alkyl phenols, for example
33、 Tergitol NP-10 orTriton X-100 and 0.5 % Na2CO3.7.7 Neutralizing BrothsGrowth media appropriate for thechallenge microorganism containing chemical agents to sup-press antibacterial activity. Alternatively, the neutralizingbroths may be of sufficient volume to reduce the concentrationof the antimicro
34、bials to below active levels. See step 11.8.7.8 Challenge Microorganisms:97.8.1 Klebsiella pneumoniae, ATCC 4352.7.8.2 Staphylococcus aureus, ATCC 6538.7.8.3 Pseudomonas aeruginosa, ATCC 15442.7.8.4 Other microorganisms, as applicable.7.9 Culture Media:7.9.1 Nutrient Agar A.7.9.2 Nutrient Agar B.7.9
35、.3 Media suitable for identification of microorganism(s)used in the study.7.9.4 Soybean casein digest medium or other suitable me-dia, with or without specific neutralizers, for recovery of thechallenge microorganism(s).7.10 Organic Soil LoadWhen an organic soil load is to beincorporated in the susp
36、ension of the challenge microorgan-ism(s), defibrinated heat-inactivated animal serum may be usedor a mixture of the following stock solutions in phosphatebuffer dilution water (pH 7.2) may be used (see 7.4).7.10.1 Add 0.5 g of tryptone to 10 mL phosphate buffer.7.10.2 Add 0.5 g of bovine serum albu
37、min (BSA) to 10 mLof phosphate buffer.7.10.3 Add 0.04 g of bovine mucin to 10 mL of phosphatebuffer.7.10.4 Prepare the solutions separately and sterilize bypassage through a 0.22 m pore diameter membrane filter,aliquote and store at either 4 6 2C or 20 6 2C for no longerthan 3 months.7.10.5 To obtai
38、n a 500 L inoculum of the challengemicroorganism, add to 340 L of the microbial suspension 25L, 100 L and 35 L of BSA, mucin and tryptone stocksolutions, respectively.NOTE 5The quality of the above materials may vary among manu-facturers or product lots. Therefore, preliminary screening of such item
39、s isrecommended to ensure compatibility with the test microorganism(s).NOTE 6The investigator should confirm the appropriate organic soilusage with the appropriate regulatory agency prior to initiating testing.8. Fabric and Spindle Preparation8.1 Scour test fabric by boiling approximately 300 g ofma
40、terial for1hin3Lofdistilled or deionized water containing1.5-g sodium carbonate and 1.5-g nonionic wetting agent.Rinse fabric, first in boiling water and then in cold water, untilall visual traces of wetting agent are removed (that is, foam-ing). Remove as much water as possible from fabric.8.2 Air
41、dry for at least 24 h at ambient room temperature.8.3 Cut scoured dry fabric into strips 2 in. (5 cm) wide andweighing 15 6 0.1 g each. For cotton fabrics, pierce one endof the 15-g test fabric strip and secure onto the outer horizontalextension of a stainless steel spindle. Wind the strip around th
42、ethree horizontal extensions with sufficient tension to obtain 12but not 13 laps while using the entire 15 6 0.1 g of fabric.Staples or a pin may be used to secure the fabric strip end.Apply additional staples to the 6th and 7th folds along onehorizontal side of the fabric bundle to create “pockets”
43、 that willsecure individual fabric swatches during tumbling. Fabricwrapped spindles may be sterilized in individual ExposureChambers. Alternatively, fabric wrapped spindles may besterilized separately from Exposure Chambers. Ensure drynessof fabric on spindles and Exposure Chambers prior to testing.
44、NOTE 7Fabric may be purchased in pre-cut strips and then scoured.8.4 Fabric carriers of approximately 1 by 1.5 in. will be cutfrom the remaining scoured fabric. A nontoxic permanentmarker may be used to place a mark on the edge of each carrier.Alternatively, attach a pin to the short side of each ca
45、rrier.Place fabric carriers in glass Petri dishes and sterilize. Ensuredryness of fabric prior to testing.8.5 For each exposure chamber, prepare at least 3 fabriccarriers and 1 fabric wrapped spindle for each active testformulation/product and control/numbers control.9. Preparation of Challenge Micr
46、oorganisms9.1 Subculture microorganism(s) on Nutrient Agar Athrough at least three daily transfers, incubating at 35 6 2C.If only one daily transfer is missed, it is not necessary to repeatthe three consecutive transfers prior to use in testing.9.2 On the day prior to testing, transfer the cells int
47、o Frenchsquare bottles containing 20 mL Nutrient Agar B. Incubate 18to24hat356 2C, agar side down.9.3 Remove growth from the French square bottles usingthree-mL dilution fluid and five sterile glass beads to suspend8Fabric #400 8-1978) obtained from Test Fabric, Inc., P.O. Box 26, West Pittson,PA 18
48、643 or supplier of fabric with comparable specifications.9DIS/TSS 13. Laundry AdditivesDisinfection and Sanitization. U.S. Environ-mental Protection Agency, Office of Pesticide Programs, April 1980.E2406043growth. The cultures will be standardized to yield approxi-mately 108colony forming units (CFU
49、) per mL of S. aureusand 109CFU/mL of K. pneumoniae and P. aeruginosa.NOTE 8The initial inoculum concentration for different challengemicroorganisms may vary and should be determined from carrier andwash water numbers control recovery (see Section 12).9.4 A soil load may be added to each inoculum (see 7.10).10. Preparation of Test Sample10.1 Prepare a sufficient volume of active test formulationand product control (at least 1 L) according to manufacturerinstructions, diluted in synthetic water with appropriate hard-ness and pre-equilibr
