1、Designation: D7225 13Standard Guide forBlood Cleaning Efficiency of Detergents and Washer-Disinfectors1This standard is issued under the fixed designation D7225; the number immediately following the designation indicates the year oforiginal adoption or, in the case of revision, the year of last revi
2、sion. 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 guide is based on a standardized test soil correlat-ing to coagulated blood suitable for screening tests and theevaluation
3、 of the cleaning efficiency of washer-disinfectorsused for reprocessing of surgical instruments. This guidestrictly deals with cleaning and does not describe any methodsthat are related to disinfection. See the Referenced DocumentsD5343, D4008, D4265, D4488, D2960, D3050, in Section 2for additional
4、information.1.2 The values stated in SI units are to be regarded asstandard. No other units of measurement are included in thisstandard.1.3 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 es
5、tablish appro-priate safety and health practices and to determine theapplicability of regulatory limitations prior to use.2. Referenced Documents2.1 ASTM Standards:2D5343 Guide for Evaluating Cleaning Performance of Ce-ramic Tile CleanersD4008 Test Method for Measuring Anti-Soil DepositionProperties
6、 of Laundry Detergents (Not Suitable for Deter-gent Ranking)D4265 Guide for Evaluating Stain Removal Performance inHome LaunderingD4488 Guide for Testing Cleaning Performance of ProductsIntended for Use on Resilient Flooring and WashableWalls (Withdrawn 2009)3D2960 Guide for Controlled Laundering Te
7、st Using Natu-rally Soiled Fabrics and Household AppliancesD3050 Guide for Measuring Soil Removal from ArtificiallySoiled Fabrics (Not Suitable for Detergent Ranking)2.2 AAMI Standards:4ANSI/AAMI ST79 Comprehensive guide to steam steriliza-tion and sterility assurance in health care facilitiesAAMI T
8、IR 12 Designing, testing, and labelling reusablemedical devices for reprocessing in health care facilities:a guide for medical device manufacturersANSI/AAMI ST81 Sterilization of medical devices infor-mation to be provided by the manufacturer for theprocessing of resterilizable medical devicesAAMI T
9、IR 30 A compendium of processes, materials, testmethods, and acceptance criteria for cleaning reusablemedical devices2.3 ISO Standards5ISO 158832 Washer-disinfectors, part 2: requirements andtests for washer-disinfectors employing thermal disinfec-tion for surgical instruments, anaesthetic equipment
10、,bowls, dishes, receivers, utensils, glassware, etc.ISO/TS 158835:2005 Washer-disinfectors, part 5: test soilsand methods for demonstrating cleaning efficacy ofwasher-disinfectors3. Summary of Guide3.1 The standardized test soil is based on a proteinousmatrix containing fibrinogen and thrombin in tw
11、o separatedcomponents. Coagulation and formation of fibrin fibers areinduced after mixing the two components.3.2 The suggested methods are based on the removal ofstandardized test soil as a result of mechanical or chemicalaction, or both, of the tested detergents or washer-disinfectors,or both. The
12、screening test provides qualitative results forcleaning efficacy. After testing the practical situation in awasher-disinfector, the end result is visually checked forimmediate evaluation. Minor residue is detected by using the1This guide is under the jurisdiction of ASTM Committee D12 on Soaps andOt
13、her Detergents and is the direct responsibility of Subcommittee D12.16 on HardSurface Cleaning.Current edition approved June 1, 2013. Published July 2013. Originally approvedin 2006. Last previous edition approved in 2006 as D7225 06. DOI: 10.1520/D7225-13.2For referenced ASTM standards, visit the A
14、STM 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.3The last approved version of this historical standard is referenced onwww.astm.org.4Available from Amer
15、ican National Standards Institute (ANSI), 25 W. 43rd St.,4th Floor, New York, NY 10036.5Available from International Organization for Standardization (ISO), 1, ch. dela Voie-Creuse, CP 56, CH-1211 Geneva 20, Switzerland, http:/www.iso.org.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C
16、700, West Conshohocken, PA 19428-2959. United States1peroxidase reaction. The quantitative test provides quantitativeresults for cleaning efficacy by providing measuring the re-moval of a known amount of protein. After placement of thetest coupon within a washer or in a beaker of water, the endresul
17、t is measured gravimetrically or by spectophotometerdetection of protein residue, or both.4. Significance and Use4.1 SignificanceDried blood represents a significant chal-lenge to cleaning surgical instruments. The water-solublecomponents of blood are easily rendered insoluble whenexposed to heat, c
18、hemical solutions, or time at room tempera-ture. The water insoluble component of blood is fibrin built upduring coagulation. These proteins bind quite readily to thesurfaces of surgical instruments making them difficult toremove even with the aid of chemical cleaning agents. Instru-ments contaminat
19、ed with blood residue after reprocessingrepresent a significant threat for infection to healthcare workersand patients. Healthcare facilities typically employ the use ofautomated instrument washers. These devices combine me-chanical action along with chemical cleaning agents in a stagedcleaning cycl
20、e designed to thoroughly clean surgical instru-ments. To function properly, these machines must be perform-ing at targeted mechanical efficiency and deliver the correctchemical cleaning agents at the correct temperature, at thecorrect dosage for the correct period of time. Manufacturers ofautomated
21、washers and manufacturers of cleaning detergentneed to evaluate the performance of their products utilizing asurrogate for surgical instruments soiled with blood. Theresults of the performance testing will be used to improveproduct design and for validation of the performance of theirproduct for var
22、ious regulatory requirements.4.2 UseThe regular, periodic use of the blood soil test is asystemic challenge to the functioning of an automated washer.To properly challenge the cleaning device, the test must beanalogous to the dried blood soil, to the stainless steelsubstrate, and to the physical bar
23、riers presented by surgicalinstruments. These physical barriers include the box lock, orpivot joint of a hinged instrument, the serrated tips, andcrevices of surgical instruments. On the test coupon, thecomponents of blood are similar to the state of dried blood oninstruments. By utilizing a grooved
24、 stainless steel coupon, thesubstrate is similar to that of stainless steel instruments. Bymounting the soiled coupon in a plastic holder the physicalbarriers represented by cracks and crevices of instruments (forexample, box locks) are represented. Users are provided withan interpretation guide tha
25、t aids them in interpreting results thatare less than optimal. For instance, failure to remove the fibrinlayer of blood soil (which is water insoluble) indicates aproblem with the chemical cleaning agent(s). Failure to evenlyremove a hemoglobin soil indicates a mechanical failure.Failure to remove a
26、ny soil indicates either a catastrophicmechanical failure, or inappropriate settings for the initial rinsestage. As a standardized challenge, the test provides a repro-ducible means for the washer manufacturer and the detergentmanufacturer to compare new designs and formulations tothose existing wit
27、hin their own product line as well as those ofothers in the market. For the purpose of submitting theirinstructions for use, the test provides a means to validate theperformance of their product with a device that is a surrogatefor the devices their products will be used to clean in thepractical set
28、ting. This validation testing can be used as part ofany necessary documentation for regulatory filings and re-cords.5. Reagents and Materials5.1 Standardized Test Soil5.1.1 Composition:5.1.1.1 Component A:400-mg albumine, bovine, protease free;400-mg hemoglobin, bovine, lyophilized; and60-mg fibrino
29、gen, bovine, lyophilized.5.1.1.2 Component B:400-mg albumine, bovine, protease free;400-mg hemoglobin, bovine, lyophilized; and12.5-NIH units thrombin, reagent grade from bovineplasma.5.1.1.3 Solvent A5.0-mL 0.4 % NaCl solution (Reagentgrade NaCl dissolved in sterile water).5.1.1.4 Solvent B5.0-mL 0
30、.4 % NaCl solution + 8.0-mmol/L CaCl2. (Reagent grade NaCl dissolved in sterilewater).5.2 PreparationComponents A and B are dissolved intheir corresponding Solvents A and B by shaking for1hatroom temperature (from 20 to 37C ) in two sealed 10-mLglass vials. Shaking by hand is acceptable and the most
31、easy/practical solution and is tested to give reliable results. Alaboratory quality shaker could be used for a more reproducibleprocess.6. Sampling, Test Specimens, and Test Units6.1 Preparation of Test Coupons6.1.1 Coupons for Screening Test6.1.1.1 Stainless steel plates (70 20 1 mm, material:1.430
32、1) are used for the screening test to evaluate the chemicalefficiency of detergents. The steel plates must be precleanedand free of any residue. They should be marked with awaterproof marker. A blind control of the finished test shouldalways be conducted: Immersion test of a test object indeminerali
33、zed water at room temperature (58 to 77F).6.1.1.2 Fifty microlitres of the dissolved Component A and50 Lof the dissolved Component B are dosed onto the surfaceof the coupons, mixed, and spread to cover approximately 4cm2. Both components are dosed on the same spot of the sameside of the plate, mixed
34、 (with pipette) and spread onto thesurface.6.1.1.3 The test coupons are then dried at room temperature(68 to 77F) and 40 to 60 % humidity for 24 h (convection maybe used to speed the drying process). Coupons needs to bestored out of reach of any chemicals or fumes (for example,disinfectants, deterge
35、nts, or solvents).7. Procedure7.1 Peroxidase Reaction for the Detection of Residue7.1.1 Test Solution:D7225 1327.1.1.1 Solution 10.1 % tetramethylbenzidine (TMB) in5 % acetic acid.7.1.1.2 Solution 23 % hydrogen peroxide solution.7.1.2 Detection of Residues:7.1.2.1 A swab is used to sample nontranspa
36、rent lumens orsurfaces. Only use a clean cotton swab that does not react withthe test solution. If surfaces are dry, a swab is moistened witha drop of water. If the surface is wet, the swab need not bepre-moistened.7.1.2.2 Activate 1 mL of Solution 1 with 100 L of Solution2 and drop the swab in the
37、liquid. Blood residue will beindicated by a blue color reaction. Visible color change willoccur at 0.1 g or greater.7.2 Screening Test for Enzymatic Detergents:7.2.1 MethodPrepare enzymatic detergent solution ac-cording to manufacturers recommendations. Enzymatic deter-gents are normally used at 0.5
38、 % (v/v) solutions. Dissolve 500L of enzymatic detergent concentrates in 100-mL flasks andfill up to volume with water of known quality (cleaningefficiency should be compared in demineralized water andwater of standardized hardness). Transfer the solution into100-mL beakers and warm up in a waterbat
39、h until the recom-mended temperature has been reached. Usually enzymaticdetergents show best blood-cleaning efficiency at 45C. Insertvertically three test coupons into the detergent solution so thatthe test soil and coupon are completely submerged. No stirring,shaking or any other means of mechanica
40、l action should beapplied. Remove the coupons after 15, 30, and 60 min, let dry,and visually inspect for remaining residues (fibrin fibers).7.3 Test for Cleaning Effciency of Washer-Disinfectors7.3.1 Method:7.3.1.1 Contaminate surgical instruments by pipette withtest soil prepared in 5.1 and let coa
41、gulate and dry. Take carethat critical parts of the instruments are contaminated, forexample, joints or lumen. Specially designed test couponsmight be used instead of real instruments.7.3.1.2 Place contaminated instruments or test coupons oninstrument trays and reprocess in the washer-disinfector wi
42、ththe recommended program for decontamination of contami-nated surgical instruments. Ensure that cannulated instrumentsare connected to the channel irrigation system of the washer.7.3.2 Detection of Residue:7.3.2.1 Peroxidase MethodInspect instruments or testcoupons for any visible residues and use
43、the peroxidasereaction described in 7.1 for detecting blood residues insideinvisible places (for example, lumens or joints)7.3.2.2 Gravimetric MethodMeasure the weight of the testcoupons with a balance capable of measuring 0.1 mg. For thispurpose, the test coupons need to be marked (see 6.1) and als
44、omeasured before the application of the test soil:cleaning efficiency 5 100 2 (m2 2 m0)(m1 2 m0) 3100 %(1)wherem0 = weight of the empty test coupon, mg,m1 = weight of the test coupon with dried test soil, mg, andm2 = weight of the test coupon after the dipping test, mg.7.3.2.3 Chemical Detection Met
45、hodElute residual organicmaterial from the surface of the test:(1) Immerse coupon in a sterile beaker containing 3 mL ofBradfords reagent.(2) Soak for 30 min at room temperature.(3) Mix solution.(4) Transfer 20 L aliquots to wells of 96 well microtitretray.(5) Test for absorbance at 595 nm for prote
46、in.8. Interpretation of Results8.1 A visual clean surface without detectable traces ofresidues is the optimum result for checking the cleaningefficacy of processes. Additional information regarding theactivity of a detergent or cleaning process can be achieved bythe amount of time needed to complete
47、ly remove all protein.Visible fibrin (colorless protein) indicates a lack of chemicalcleaning efficacy while visible hemoglobin (red protein resi-due) indicates a lack of water/detergent coverage in a washer.NOTE 1Pure water will give a cleaning efficiency of approximately95 %, leaving only the fibr
48、in fibers behind. An active detergent needs toclean the test coupons completely, indicating a pass/fail result.9. Keywords9.1 blood cleaning efficiency; detergents; washer-disinfectorsASTM International takes no position respecting the validity of any patent rights asserted in connection with any it
49、em mentionedin this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the riskof infringement of such rights, are entirely their own responsibility.This standard is subject to revision at any time by the responsible technical committee and must be reviewed every five years andif not revised, either reapproved or withdrawn. Your comments are invited either for revision of this standard or for additional standardsand should be addressed to ASTM International Headquarters. Your co
