ASTM E1174-00
(Test Method)Standard Test Method for Evaluation of the Effectiveness of Health Care Personnel or Consumer Handwash Formulations
Standard Test Method for Evaluation of the Effectiveness of Health Care Personnel or Consumer Handwash Formulations
SCOPE
1.1 This test method is designed to determine the effectiveness of antimicrobial handwashing agents for the reduction of transient microbial flora when used in a handwashing procedure.
1.2 A knowledge of microbiological techniques is required for these procedures.
1.3 In this test method metric units are used for all applications, except for distance in which case inches are used and metric units follow in parentheses.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. For more specific precautionary statements see Note 1.
1.4 This method may be used to evaluate topical antimicrobial handwash formulations.
1.5 Performance of this procedure requires the knowledge of regulations pertaining to the protection of human subjects.
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Designation:E1174–00
Standard Test Method for
Evaluation of the Effectiveness of Health Care Personnel or
Consumer Handwash Formulations
This standard is issued under the fixed designation E 1174; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
1. Scope microbial contaminant. It may also be referred to as a marker
organism, bacterial simulant, or bacterial contaminant.
1.1 This test method is designed to determine the effective-
3.1.2 resident microorganisms—microorganisms that live
ness of antimicrobial handwashing agents for the reduction of
and multiply on the skin, forming a permanent population.
transient microbial flora when used in a handwashing proce-
3.1.3 transient microorganisms—organisms from the envi-
dure.
ronment that contaminate but do not normally colonize the
1.2 A knowledge of microbiological techniques is required
skin.
for these procedures.
3.1.4 active ingredient—a substance added to a formulation
1.3 In this test method metric units are used for all applica-
specifically for the inhibition or inactivation of microorgan-
tions, except for distance in which case inches are used and
isms.
metric units follow in parentheses.
3.1.5 test formulation—a formulation which incorporates
1.4 This standard does not purport to address all of the
antimicrobial ingredient(s).
safety concerns, if any, associated with its use. It is the
3.1.6 neutralization—a process which results in quenching
responsibility of the user of this standard to establish appro-
the antimicrobial activity of a test material. This may be
priate safety and health practices and determine the applica-
achieved through dilution of the test material(s) to reduce the
bility of regulatory limitations prior to use. For more specific
antimicrobial activity, or through the use of chemical agents,
precautionary statements see Note 1.
called neutralizers, to eliminate antibacterial activity.
1.5 This method may be used to evaluate topical antimicro-
3.1.7 cleansing wash—a non-antimicrobial wash intended
bial handwash formulations.
to remove gross soil or residues from the hands of the panelists
1.6 Performance of this procedure requires the knowledge
prior to the conduct of the study and as noted throughout the
of regulations pertaining to the protection of human subjects.
study. This may also be referred to as a cosmetic wash.
2. Referenced Documents
3.1.8 healthcare personnel handwash—a cleanser or water-
less agent intended to reduce transient bacteria on the hands.
2.1 ASTM Standards:
E 1054 Practices for Evaluating Inactivators of Antimicro-
4. Summary of Test Method
bial Agents Used in Disinfectant, Sanitizer, Antiseptic, or
4.1 This test method is conducted on a group of volunteer
Preserved Products
panelists who have refrained from using topical antimicrobial
3. Terminology formulations for at least one week prior to the initiation of the
test.Activity of the test material is measured by comparing the
3.1 Definitions:
number of test organisms recovered from artificially contami-
3.1.1 test organism—an applied inoculum of an organism
nated hands after use of a handwashing formulation to the
that has characteristics which allow it to be readily identified.
number recovered from contaminated hands not exposed to the
The test organism is used to simulate a transient topical
test formulation. The method describes specific procedures to
be followed using Serratia marcescens as the test organism.
This test method is under the jurisdiction of ASTM Committee E35 on
The activity of the test material may be measured following a
Pesticides and Alternative Control Agents and is the direct responsibility of
single wash and multiple washes in a single clay using a
Subcommittee E35.15 Antimicrobial Agents.
neutralization recovery method.
Current edition approved August 10, 2000. Published November 2000. Origi-
4.2 An alternative test organism is Escherichia coli. Culture
nally published as E 1174 – 87. Last previous edition E 1174 – 94.
Federal Register, Vol 46, No. 17, Jan. 27, 1991.
media and incubation conditions appropriate for this organism
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
shouldbeemployed.Theinvestigatorshouldalsobeawarethat
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
there may be health risks associated with the use of this
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E1174–00
organism and precautions similar to those referenced in Note 1 7.7 Sampling Solution—Dissolve 0.4 g KH PO , 10.1 g
2 4
should be undertaken. Na HPO and 1.0 g isooctylphenoxypolyethoxyethanol and
with appropriately validated neutralizers in 1-L distilled water.
5. Significance and Use
Adjust pH to 7.8 with 0.1 N HCl or 0.1 N NaOH. Dispense so
that final volume after sterilization is 75 ml, sterilized at
5.1 The procedure may be used to test the effectiveness of
121°C.
antimicrobial handwashing agents. The test formulations may
7.8 Dilution Fluid—Sterile Butterfield’s Buffer or other
be designed for frequent use to reduce the transient bacterial
suitable diluent, adjusted to pH 7.2 with effective neutralizer
flora on hands.
forthetestmaterial.AdjustpHwith0.1 NHClor0.1 NNaOH.
6. Apparatus See Test Methods E 1054.
7.9 Agar—Soybean-casein digest agar, or other solid media
6.1 Colony Counter—Any of several types may be used, for
appropriately validated to support growth of the test organism
example, Quebec Colony Counter.
with appropriate neutralizers if needed.
6.2 Incubator—Any incubator capable of maintaining the
7.10 Broth—Soybean-casein digest broth or other liquid
following temperatures: S. marcescens (25 6 2°C) or E. coli
media appropriate to support growth of the test organism.
(35 6 2°C). This temperature is required to ensure pigment
production for S. marcescens.
8. Test Organism
6.3 Sterilizer—Any suitable steam sterilizer capable of
8.1 Serratia marcescens (ATCC 14756) is to be used as the
producing the conditions of sterilization is acceptable.
test organism. This is a strain having stable pigmentation at
6.4 Timer (Stop-clock)—One that can be read for minutes
25°C.
and seconds.
8.2 Escherichia coli (ATCC 11229) is an alternative test
6.5 Handwashing Sink—A sink of sufficient size to permit
organism.When E. coli is used, the plating agar should include
panelists to wash without touching hands to sink surface or
a suitable indicator (e.g. MUG ).
other panelists.
6.5.1 Water faucet(s)—To be located above the sink at a
NOTE 1—Warning: Theapplicationofmicroorganismstotheskinmay
height which permits the hands to be held higher than the involve a health risk. Prior to applying the test organism to the skin, the
antibiotic sensitivity profile of the strain should be determined. If the
elbow during the washing procedure.
strain is not susceptible to gentamicin, do not use it. If an infection occurs,
6.6 Tap Water Temperature Regulator and Temperature
the antibiotic sensitivity profile should be made available to the attending
Monitor—To monitor and regulate water temperature of 40 6
clinician.
2°C.
Following the subject’s last contamination and wash with the formula-
tion, the subject’s hands are to be sanitized by scrubbing with 70%
7. Reagents and Materials
isopropanol solution or equivalent. The purpose of this alcohol scrub is to
destroy residual test organisms on the skin.
7.1 Bacteriological Pipettes—10.0 and 2.2-mL or 1.1-mL
capacity.
8.3 Preparation of Test Organism Suspension
7.2 Water Dilution Bottles—Any sterilizable glass container 8.3.1 S. marcescens—A homogeneous culture is used to
having a 150–200 mL capacity and tight closures may be inoculate the hands.The stock culture should be at least two 24
used.
hour broth transfers from the original ATCC culture, but there
7.3 Erlenmeyer Flask—2-L capacity for culturing test or- should be no more than 5 transfers removed from the ATCC
ganism.
culture. From the stock culture of Serratia marcescens (ATCC
7.4 Cleansing Wash—A mild, non-antimicrobial solid or 14756) inoculate the appropriate volume of soybean-casein
liquid soap. (The investigator may choose to use the product
digest broth (7.10) with 0.1 milliliter of stock culture of S.
vehicle.) marcescens/100mLs of broth to yield the volume necessary to
7.5 Test Material—Directions for use of the test material
complete the study. Incubate for 24 6 4 h at 25°C 6 2°C.
may be utilized. If directions are not available, use directions Broth should develop a red pigment.
provided in this test method.
8.3.2 E. coli—A homogeneous culture is used to inoculate
7.6 Gloves—Loose-fitting, unlined, powder-free gloves thehands,thestockcultureshouldbeatleasttwo24hourbroth
which possess no antimicrobial properties, or equivalent.
transfers from the original ATCC culture, but no more than 5
(Plastic bags with low bioburden may be used in place of transfers removed from the ATCC culture. From the stock
gloves.)
Triton X-100, Rohm and Haas Co., Philadelphia, PA.
Peterson, A.F., “The Microbiology of the Hands: Evaluating the Effects of the
Presterilized/disposable bacteriological pipettes are available from most local Surgical Scrubs,” Developments in Industrial Microbiology, Vol 14, pp. 125–130,
laboratory supply houses. 1973.
5 9
Milk dilution bottles of 160-mL capacity having a screw-cap closure are Horowitz, W. (Ed.) 1980. Offıcial Methods of Analysis of the AOAC, 13th Ed.,
availablefromCorningGlassCo.,KimbleGlassCo.ormostlocallaboratorysupply Sec. 46.013 (m), p. 825.Assoc. of Off.Anal. Chemists, Washington, D.C. 1018 pp.
houses. United States Pharmacopeia XXII: United States Pharmacopeial Convention,
AsuitableglovewouldbePharmaseal8873C,(sterile)FlexamLatexProcedure Inc., Rockville, MD, Chapter entitled “Microbial Limits Test.” The MUG (4-
Glove from American Pharmaseal Laboratories, Glendale, CA 91209. A zone of methylumbelliferyl-b-D-gluconride)substrateishydrolyzedby b-D-gluconridaseto
inhibition test such as AATCC Test Method 90-1965 may be used to evaluate yield a fluorescent end product, 4-methylumbelliferone. b-D-gluconridase is pos-
antimicrobial properties of gloves, AATCC Test Methods, American Association of sessed by E. coli (ATCC 11229). MUG is incorporated into the appropriate growth
Textile Chemists and Colorist, 1968 Technical Manual, Section B-75. medium at 0.05 grams/L.
E1174–00
NOTE 2—The hands may still be wet after the 90 seconds.
culture of Escherichia coli
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