ASTM F1671/F1671M-22

Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Blood-Borne Pathogens Using Phi-X174 Bacteriophage Penetration as a Test System

SIGNIFICANCE AND USE
5.1 This test method is based on Test Method F903 for measuring resistance of chemical protective clothing materials to penetration by liquids. This test method is normally used to evaluate specimens from individual finished items of protective clothing and individual samples of materials that are candidates for items of protective clothing.  
5.1.1 Finished items of protective clothing include gloves, arm shields, aprons, gowns, coveralls, hoods, and boots.  
5.1.2 The phrase “specimens from finished items” encompasses seamed and other discontinuous regions, as well as the usual continuous regions of protective clothing items.  
5.2 It is known that body fluids penetrating protective clothing materials are likely to carry microbiological contaminants; however, visual detection methods are not sensitive enough to detect minute amounts of liquid containing microorganisms (1-3).7 This test method uses media containing Phi-X174 Bacteriophage. The visual detection technique of this test method is supplemented with a biologically based assay capable of detecting virus under the specified test conditions.  
5.3 Test Method F1670/F1670M allows the screening of protective clothing materials for resistance to penetration with synthetic blood as a challenge liquid. Test Method F1670/F1670M uses the same penetration test cell and technique, but exposes material specimens to synthetic blood with visual detection of liquid penetration. Materials passing Test Method F1670/F1670M should then be tested against bacteriophage penetration using this test method to verify performance.  
5.4 This test method has been specifically designed for measuring penetration of a surrogate microbe for Hepatitis (B and C) and the Human Immunodeficiency Viruses. The surrogate, Phi-X174 Bacteriophage, used in this test method is similar to HCV in size and shape but also serves as a surrogate for HBV and HIV. Inferences about protection from other pathogens must be assessed on a case-by-case basi...
SCOPE
1.1 This test method is used to measure the resistance of materials used in protective clothing to penetration by blood-borne pathogens using a surrogate microbe under conditions of continuous liquid contact. Protective clothing material pass/fail determinations are based on the detection of viral penetration.  
1.1.1 This test method is not always effective in testing protective clothing materials having thick, inner liners which readily absorb the liquid assay fluid.  
1.2 This test method does not apply to all forms or conditions of blood-borne pathogen exposure. Users of the test method should review modes for worker/clothing exposure and assess the appropriateness of this test method for their specific applications.  
1.3 This test method has been specifically defined for modeling the viral penetration of Hepatitis (B and C) and Human Immunodeficiency Viruses transmitted in blood and other potentially infectious body fluids. Inferences for protection from other pathogens must be assessed on a case-by-case basis.  
1.4 This test method addresses only the performance of materials or certain material constructions (for example, seams) used in protective clothing and determined to be viral resistant. This test method does not address the design, overall construction and components, or interfaces of garments or other factors which may affect the overall protection offered by the protective clothing.  
1.5 The values stated in SI units or in other units shall be regarded separately as standard. The values stated in each system must be used independently of the other, without combining values in any way.  
1.6 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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.7 This i...

General Information

• Status: Published
• Publication Date: 31-Dec-2021
• ICS: 11.140 - Hospital equipment
• ICS: 13.340.10 - Protective clothing
• Technical Committee: F23 - Personal Protective Clothing and Equipment
• Drafting Committee: F23.40 - Biological

Relations

• Refers: ASTM F1670/F1670M-24 - Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Synthetic Blood
• Effective Date: 01-Feb-2024
• Refers: ASTM E171/E171M-11(2020) - Standard Practice for Conditioning and Testing Flexible Barrier Packaging
• Effective Date: 01-May-2020
• Refers: ASTM D1777-96(2019) - Standard Test Method for Thickness of Textile Materials
• Effective Date: 01-Jul-2019
• Refers: ASTM F903-18 - Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Liquids
• Effective Date: 01-Jul-2018
• Refers: ASTM F1670/F1670M-17a - Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Synthetic Blood
• Effective Date: 25-Oct-2017
• Refers: ASTM D3776/D3776M-09a(2017) - Standard Test Methods for Mass Per Unit Area (Weight) of Fabric
• Effective Date: 15-Jul-2017
• Refers: ASTM F1670/F1670M-17 - Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Synthetic Blood
• Effective Date: 01-Jul-2017
• Refers: ASTM F903-17 - Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Liquids
• Effective Date: 01-Feb-2017
• Refers: ASTM E171/E171M-11(2015) - Standard Practice for Conditioning and Testing Flexible Barrier Packaging
• Effective Date: 01-Oct-2015
• Refers: ASTM D1777-96(2015) - Standard Test Method for Thickness of Textile Materials
• Effective Date: 01-Jul-2015
• Refers: ASTM F1670/F1670M-08(2014)e1 - Standard Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Synthetic Blood
• Effective Date: 15-Jun-2014
• Refers: ASTM D3776/D3776M-09a(2013) - Standard Test Methods for Mass Per Unit Area (Weight) of Fabric
• Effective Date: 01-Jul-2013
• Refers: ASTM D3862-13 - Standard Test Method for Retention Characteristics of 0.2-&#181m Membrane Filters Used in Routine Filtration Procedures for the Evaluation of Microbiological Water Quality
• Effective Date: 15-Feb-2013
• Refers: ASTM E171/E171M-11 - Standard Practice for Conditioning and Testing Flexible Barrier Packaging
• Effective Date: 15-Nov-2011
• Refers: ASTM D1777-96(2011)e1 - Standard Test Method for Thickness of Textile Materials
• Effective Date: 01-May-2011

Overview

ASTM F1671/F1671M-22 is the internationally recognized standard test method for evaluating the resistance of protective clothing materials to penetration by blood-borne pathogens. Developed by ASTM International, this method specifically utilizes the Phi-X174 bacteriophage as a surrogate microbe to simulate viral penetration under continuous liquid contact conditions. This procedure is essential for manufacturers and end-users to ensure that protective clothing offers effective barriers against infectious agents such as Hepatitis B and C viruses (HBV, HCV), and Human Immunodeficiency Virus (HIV). The standard is commonly referenced in healthcare, emergency response, and other occupational safety settings where exposure risk to blood and body fluids exists.

Key Topics

• Purpose and Use: The standard provides a systematic approach for determining the viral penetration resistance of protective clothing materials, including gloves, aprons, gowns, coveralls, and related items. Both finished items and material specimens-covering seams and discontinuous regions-are evaluated.
• Surrogate Microbe: Phi-X174 bacteriophage is used as a model for blood-borne viruses due to its similar size and shape to pathogens like HBV, HCV, and HIV, providing a realistic but safe test system.
• Testing Protocol: The method involves exposing samples to a bacteriophage-containing solution under controlled pressure and assessing for viral penetration using sensitive biological assays. This is significantly more precise than visual detection alone, which may not identify minute penetrations.
• Limitations: The standard cautions that it may not be suitable for materials with thick, absorbent inner liners and does not address all potential exposure scenarios, garment designs, or forms of degradation over time.
• Regulatory Alignment: ASTM F1671/F1671M-22 is cited in regulations, such as OSHA’s standards on occupational exposure to blood-borne pathogens, reinforcing its importance for compliance and worker safety.

Applications

The ASTM F1671/F1671M-22 standard is critical across multiple industries where protection from blood-borne pathogens is paramount:

• Healthcare: Used for certifying the barrier effectiveness of surgical gowns, gloves, drapes, and other personal protective apparel to support infection control procedures.
• First Responders and Laboratory Staff: Ensures that protective clothing worn in high-risk environments effectively blocks virus transmission through body fluids.
• Product Development & Quality Assurance: Manufacturers and suppliers employ this standard in R&D, production, and quality testing phases to validate new textile constructions or assess ongoing performance.
• Compliance and Procurement: Organizations use ASTM F1671/F1671M-22 to select appropriate protective apparel that meets industry and regulatory requirements, ensuring safety and legal conformity.

Related Standards

Several industry and ASTM standards complement ASTM F1671/F1671M-22 for a comprehensive approach to protective clothing assessment:

• ASTM F903: Resistance of Materials Used in Protective Clothing to Penetration by Liquids.
• ASTM F1670/F1670M: Resistance of Materials Used in Protective Clothing to Penetration by Synthetic Blood.
• ISO 2859-1: Sampling procedures for inspection by attributes.
• ANSI/ASQ Z1.4: Sampling Procedures and Tables for Inspection by Attributes.
• OSHA 29 CFR 1910.1030: Occupational Exposure to Bloodborne Pathogens.
• ASTM D1331, D1777, D3776, D3862: Supporting test methods for measuring surface tension, material thickness, weight, and retention characteristics relevant to the performance testing of protective materials.

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By applying ASTM F1671/F1671M-22, organizations demonstrate a commitment to high standards of protective clothing performance, supporting safer work environments and compliance with infection prevention regulations. This standard is pivotal for anyone involved in evaluating, manufacturing, or purchasing protective apparel intended to guard against blood-borne pathogen exposure.

Standards Content (Sample)

This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: F1671/F1671M − 22
Standard Test Method for
Resistance of Materials Used in Protective Clothing to
Penetration by Blood-Borne Pathogens Using Phi-X174
Bacteriophage Penetration as a Test System
ThisstandardisissuedunderthefixeddesignationF1671/F1671M;thenumberimmediatelyfollowingthedesignationindicatestheyear
of original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.
A superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
Workers, primarily those in the health care profession, involved in treating and caring for
individualsinjuredorsick,canbeexposedtobiologicalliquidscapableoftransmittingdisease.These
diseases, which may be caused by a variety of microorganisms, can pose significant risks to life and
health.This is especially true of blood-borne viruses which cause Hepatitis (Hepatitis BVirus (HBV)
and Hepatitis C Virus (HCV)) and Acquired Immune Deficiency Syndrome (AIDS) (Human
Immunodeficiency Virus (HIV)). Since engineering controls can not eliminate all possible exposures,
attention is placed on reducing the potential of direct skin contact through the use of protective
clothing that resists penetration (29 CFR Part 1910.1030). This test method was developed to assess
theeffectivenessofmaterialsusedinprotectiveclothingforprotectingtheweareragainstcontactwith
blood-bornepathogensusingasurrogatemicrobesuspendedinabodyfluidsimulantunderconditions
of continuous contact.
1. Scope other potentially infectious body fluids. Inferences for protec-
tion from other pathogens must be assessed on a case-by-case
1.1 This test method is used to measure the resistance of
basis.
materials used in protective clothing to penetration by blood-
bornepathogensusingasurrogatemicrobeunderconditionsof
1.4 This test method addresses only the performance of
continuousliquidcontact.Protectiveclothingmaterialpass/fail
materials or certain material constructions (for example,
determinations are based on the detection of viral penetration.
seams) used in protective clothing and determined to be viral
1.1.1 This test method is not always effective in testing
resistant. This test method does not address the design, overall
protective clothing materials having thick, inner liners which
construction and components, or interfaces of garments or
readily absorb the liquid assay fluid.
otherfactorswhichmayaffecttheoverallprotectionofferedby
1.2 This test method does not apply to all forms or condi-
the protective clothing.
tions of blood-borne pathogen exposure. Users of the test
1.5 The values stated in SI units or in other units shall be
methodshouldreviewmodesforworker/clothingexposureand
regarded separately as standard. The values stated in each
assess the appropriateness of this test method for their specific
system must be used independently of the other, without
applications.
combining values in any way.
1.3 This test method has been specifically defined for
1.6 This standard does not purport to address all of the
modeling the viral penetration of Hepatitis (B and C) and
Human Immunodeficiency Viruses transmitted in blood and safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro-
priate safety, health, and environmental practices and deter-
ThistestmethodisunderthejurisdictionofASTMCommitteeF23onPersonal
mine the applicability of regulatory limitations prior to use.
ProtectiveClothingandEquipmentandisthedirectresponsibilityofSubcommittee
F23.40 on Biological.
1.7 This international standard was developed in accor-
Current edition approved Jan. 1, 2022. Published January 2022. Originally
dance with internationally recognized principles on standard-
approved in 1995. Last previous edition approved in 2013 as F1671/F1671M–13.
DOI: 10.1520/F1671_F1671M-22. ization established in the Decision on Principles for the
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F1671/F1671M − 22
Development of International Standards, Guides and Recom- 3.1.4 assay fluid, n—a sterile liquid used to wash the test
mendations issued by the World Trade Organization Technical specimen surface to determine microbiological penetration.
Barriers to Trade (TBT) Committee.
3.1.4.1 Discussion—In this test method, the assay fluid is
bacteriophage nutrient broth and the microorganism is the
2. Referenced Documents
Phi-X174 Bacteriophage. The assay fluid is used to wash the
Phi-X174 Bacteriophage from the normal inside surface of the
2.1 ASTM Standards:
test specimen.
D1331Test Methods for Surface and Interfacial Tension of
Solutions of Paints, Solvents, Solutions of Surface-Active
3.1.5 bacteriophage, n—a type of virus which infects bac-
Agents, and Related Materials
teria.
D1777Test Method for Thickness of Textile Materials
3.1.5.1 Discussion—In this test method, the bacteriophage
D3776/D3776MTest Methods for Mass Per Unit Area
is Phi-X174.The Phi-X174 Bacteriophage is not pathogenic to
(Weight) of Fabric
humans, but serves to simulate viruses that are pathogenic to
D3862Test Method for Retention Characteristics of 0.2-µm
humans.
Membrane Filters Used in Routine Filtration Procedures
3.1.6 blood-borne pathogen, n—an infectious bacterium or
for the Evaluation of Microbiological Water Quality
virus, or other disease-inducing microbe carried in blood or
E105Guide for Probability Sampling of Materials
other potentially infectious body fluids.
E171/E171MPractice for Conditioning andTesting Flexible
Barrier Packaging
3.1.6.1 Discussion—Forthepurposeofthistestmethod,the
F903Test Method for Resistance of Materials Used in primary blood-borne pathogens include Hepatitis B Virus
Protective Clothing to Penetration by Liquids
(HBV), Hepatitis C Virus (HCV), and Human Immunodefi-
F1670/F1670MTest Method for Resistance of Materials ciencyVirus (HIV). Other microorganisms must be considered
Used in Protective Clothing to Penetration by Synthetic
on a case-by-case basis.
Blood
3.1.7 body fluid, n—any liquid produced, secreted, or ex-
2.2 Military Standard:
creted by the human body.
MIL-STD-105Sampling Procedures and Tables for Inspec-
3.1.7.1 Discussion—In this test method, body fluids include
tion by Attributes
those liquids potentially infected with blood-borne pathogens,
2.3 ANSI/ASQ Standard:
including, but not limited to: blood, semen, vaginal secretions,
ANSI/ASQ Z1.4Sampling Procedures and Tables for In-
cerebrospinal fluid, synovial fluid, peritoneal fluid, amniotic
spection by Attributes
fluid, saliva in dental procedures, any body fluid that is visibly
2.4 ISO Standard:
contaminated with blood, and all body fluids in situations
ISO 2859-1Sampling Plans for Inspection by Attributes
where it is difficult or impossible to differentiate between body
fluids (see section 29 CFR Part 1910.1030).
2.5 OSHA Standard:
29 CFR Part 1910.1030Bloodborne Pathogens
3.1.8 body fluid simulant, n—aliquidwhichisusedtoactas
a model for human body liquids.
3. Terminology
3.1.8.1 Discussion—In this test method, the body fluid
3.1 Definitions:
simulant is bacteriophage nutrient broth, which is intended as
3.1.1 agar, n—a semisolid culture medium used to support
a model for human body liquids as it approximates the lower
the growth of bacteria and other microorganisms.
end of the surface tension range for blood and body fluids
3.1.2 aseptic, adj—sterile, free from viable microbiological (excluding saliva), 0.042 6 0.002 N/m.
contamination.
3.1.9 challenge suspension, n—a liquid containing an agent
3.1.3 assay, n—analysis of a mixture to determine the
that is used to test the penetration resistance of materials.
presence or concentration of a particular component.
3.1.9.1 Discussion—In this test method, the challenge sus-
3.1.3.1 Discussion—In this test method, the component
pension is the bacteriophage challenge suspension, a nutrient
being analyzed is a microorganism, Phi-X174 Bacteriophage.
broth containing the Phi-X174 Bacteriophage.
3.1.10 lawn, n—as in microbiology, a cloudy, uniform
2 growth of bacteria in a thin layer of top agar in a petri dish.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
3.1.10.1 Discussion—Inthistestmethod,E.coliC.hasbeen
Standards volume information, refer to the standard’s Document Summary page on
selected as the bacterium used to produce the lawn.
the ASTM website.
Available from Standardization Documents Order Desk, DODSSP, Bldg. 4,
3.1.11 lysis, n—the disintegration or destruction of whole
Section D, 700 Robbins Ave., Philadelphia, PA 19111-5098, http://
bacterial cells.
dodssp.daps.dla.mil.
Available from American Society for Quality (ASQ), 600 N. Plankinton Ave.,
3.1.11.1 Discussion—In this test method, the lysis of the
Milwaukee, WI 53203, http://www.asq.org.
hostbacteria, E. coli C.,iscausedbyPhi-X174Bacteriophage.
Available fromAmerican National Standards Institute (ANSI), 25 W. 43rd St.,
4th Floor, New York, NY 10036, http://www.ansi.org.
3.1.12 medium (plural, media), n—a nutrient system for the
AvailablefromU.S.GovernmentPrintingOfficeSuperintendentofDocuments,
cultivation of cells or organisms, and especially bacteria.
732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
www.access.gpo.gov. 3.1.12.1 Discussion—In this test method, the term media is
F1671/F1671M − 22
used to describe mixtures compounded to support the growth 3.1.23.1 Discussion—In this test method, viral penetration
of specific microorganisms; for example, bacteriophage nutri- is used to describe the physical translocation of the Phi-X174
ent broth, top agar. Bacteriophage through closures, seams, pores, and pinholes or
other imperfections in materials used in protective clothing.
3.1.13 morphology, n—theformandstructureofaparticular
3.1.24 viral resistant, adj—referring to materials which
organism.
impede viral penetration under specified laboratory test condi-
3.1.14 nutrient broth, n—a liquid medium.
tions and detection methods.
3.1.14.1 Discussion—In this test method, the nutrient broth
3.1.24.1 Discussion—In this test method, protective cloth-
is the bacteriophage nutrient broth which is used to culture the
ing materials which demonstrate “pass” results are considered
host bacteria, E. coli C., and to aid in manipulating the
to be resistant to viral penetration.
Phi-X174 Bacteriophage through the various stages of the
3.1.25 virus, n—a minute infectious agent which lacks
procedure,suchassuspendingthePhi-X174Bacteriophagefor
independent metabolism and is only able to replicate within a
challenging the test material in the penetration cell, assaying
living host cell.
thenormalinsidetestmaterialsurfaceand,ifrequired,making
dilutions of the assay fluid for plating.
4. Summary of Test Method
3.1.15 penetration, n—the movement of matter through
4.1 Aspecimen is subjected to a nutrient broth containing a
closures, porous materials, seams, and pinholes or other
virus for a specified time and pressure sequence.
imperfections in protective clothing on a nonmolecular level.
4.2 Visual detection of penetration is supplemented with an
3.1.15.1 Discussion—For this test method, the specific mat-
assay procedure that will detect viable viruses which penetrate
ter is a bacteriophage challenge suspension containing nutrient
the material even when liquid penetration is not visible. Any
broth with Phi-X174 Bacteriophage.
evidence of viral penetration for a test specimen constitutes
3.1.16 plaque, n—as in virology,avisible,cleararea,which
failure.
is theoretically the result of the infection and lysis of host cells
by a single viable virus.
5. Significance and Use
3.1.16.1 Discussion—In this test method, the term plaque is
5.1 This test method is based on Test Method F903 for
used to describe a visible, clear area in the lawn of E. coli C.
measuring resistance of chemical protective clothing materials
in top agar, which is theoretically the result of a single viable
to penetration by liquids. This test method is normally used to
Phi-X174, where the bacteria have been destroyed by bacte-
evaluatespecimensfromindividualfinisheditemsofprotective
riophage infection and lysis.
clothingandindividualsamplesofmaterialsthatarecandidates
3.1.17 plaque forming unit (PFU), n—a virus particle ca- for items of protective clothing.
pable of producing plaques by infecting and lysing bacteria in
5.1.1 Finished items of protective clothing include gloves,
a lawn in top agar. arm shields, aprons, gowns, coveralls, hoods, and boots.
5.1.2 The phrase “specimens from finished items” encom-
3.1.18 plate, n—as in microbiology, a petri dish containing
passes seamed and other discontinuous regions, as well as the
culture medium.
usual continuous regions of protective clothing items.
3.1.19 protective clothing, n—an item of clothing that is
5.2 It is known that body fluids penetrating protective
specifically designed and constructed for the intended purpose
clothing materials are likely to carry microbiological contami-
of isolating all or part of the body from a potential hazard; or,
nants; however, visual detection methods are not sensitive
isolating the external environment from contamination by the
enough to detect minute amounts of liquid containing micro-
wearer of the clothing.
organisms (1-3). This test method uses media containing
3.1.19.1 Discussion—In this test method, materials used in
Phi-X174 Bacteriophage. The visual detection technique of
protectiveclothingarebeingevaluated.Thepotentialhazardof
this test method is supplemented with a biologically based
contact with blood-borne pathogens is being simulated.
assay capable of detecting virus under the specified test
3.1.20 sterile, adj—free from viable microorganisms.
conditions.
3.1.21 surrogate microbe, n—a microorganism which is
5.3 Test Method F1670/F1670M allows the screening of
used to act as a simulant for other microorganisms which are
protective clothing materials for resistance to penetration with
pathogenic to humans.
synthetic blood as a challenge liquid. Test Method F1670/
3.1.21.1 Discussion—In this test method, the surrogate mi-
F1670M uses the same penetration test cell and technique, but
crobe is the Phi-X174 Bacteriophage, intended as a model for
exposes material specimens to synthetic blood with visual
HCV and to simulate both HBV and HIV.
detection of liquid penetration. Materials passing Test Method
3.1.22 titer, n—the quantity of a substance required to react F1670/F1670M should then be tested against bacteriophage
with,ortocorrespondto,agivenamountofanothersubstance.
penetration using this test method to verify performance.
3.1.22.1 Discussion—In this test method, titer is used to
5.4 This test method has been specifically designed for
describetheconcentrationofviablebacteriophageasmeasured
measuring penetration of a surrogate microbe for Hepatitis (B
in plaque forming units per millilitre (PFU/mL).
3.1.23 viral penetration, n—the penetration of a material by
Theboldfacenumbersinparenthesesrefertothelistofreferencesattheendof
a virus. this standard.
F1671/F1671M − 22
and C) and the Human Immunodeficiency Viruses. The following an appropriate preconditioning technique represen-
surrogate, Phi-X174 Bacteriophage, used in this test method is tative of the expected conditions of use.
similar to HCVin size and shape but also serves as a surrogate
5.8 This test method involves a sensitive assay procedure
for HBV and HIV. Inferences about protection from other
for determining protective clothing material resistance to
pathogens must be assessed on a case-by-case basis.
penetration by a surrogate microbe. Because of the length of
5.5 PartoftheprotocolinProceduresAandBinTable1for
time required to complete this method, it may not be suitable
exposing the protective clothing material specimens to the
for use as a material or protective clothing quality control or
Phi-X174 Bacteriophage challenge suspension involves pres-
quality assurance procedure.
surization of the penetration cell to 13.8 kPa [2 psig]. This
5.9 Ifthisprocedureisusedforqualitycontrolortosupport
hydrostatic pressure has been documented to discriminate
broad product claims concerning the viral-resistant properties
betweenprotectiveclothingmaterialperformanceandcorrelate
of materials used in protective clothing, proper statistical
with visual penetration results that are obtained with a human
design and analysis of larger data sets than those specified in
factors validation (4). Some studies, however, suggest that
thistestmethodshouldbeperformed. Examplesofacceptable
mechanical pressures exceeding 345 kPa [50 psig] can occur
sampling plans can be found in MIL-STD-105, ANSI/ASQ
during actual clinical use (5, 6). Therefore, it is important to
Z1.4, and ISO 2859-1.
understand that this test method does not simulate all the
physical stresses and pressures that might be exerted on
5.10 This test method requires a working knowledge of
protective clothing materials during actual use.
basic microbiological techniques (9).
5.6 Medical protective clothing materials are intended to be
6. Apparatus and Materials
a barrier to blood, body fluids, and other potentially infectious
materials. Many factors can affect the wetting and penetration
6.1 Apparatus:
characteristics of body fluids, such as surface tension,
6.1.1 Thickness Gauge, suitable for measuring thickness to
viscosity,andpolarityofthefluids,aswellasthestructureand
the nearest 0.02 mm [or nearest 0.001 in.], as specified in Test
relative hydrophilicity or hydrophobicity of the materials. The
Method D1777, to determine the thickness of each protective
surface tension range for blood and body fluids (excluding
clothing material specimen tested. (Optional.)
saliva) is approximately 0.042 to 0.060 N/m (7). To help
10,11
6.1.2 Penetration Test Cell, torestrainthetestspecimen
simulate the wetting characteristics of blood and body fluids,
during contact with the pressurized Phi-X174 Bacteriophage
the surface tension of the Phi-X174 Bacteriophage challenge
challenge suspension. In the test cell, the specimen acts as a
suspension is adjusted to approximate the lower end of this
partition separating the bacteriophage challenge suspension
surface tension range. This is accomplished by adding surfac-
from the viewing side of the penetration cell. The capacity of
tant to the Phi-X174 Bacteriophage nutrient broth. The result-
thecellbodyisapproximately60mL[2.0oz]forthechallenge
ing surface tension of the Phi-X174 Bacteriophage challenge
suspension. A flange cover, with an open area to allow visual
suspension is approximately 0.042 6 0.002 N/m.
observationandassayofthespecimen,andatransparentcover
5.7 Testing prior to degradation by physical, chemical, and
areincluded.Thecellbodyhasatopportforfillingandadrain
thermal stresses which could negatively impact the perfor-
valvefordrainingthepenetrationtestcell.Otheritems,suchas
mance of the protective material could lead to a false sense of
afittingtoallowattachmentoftheairlinetothetopportinthe
security. Additional tests should be considered that assess the
cell body, gaskets, and the retaining screen will also be
impact of storage conditions and shelf life on disposable
required. Specifications for the penetration cell are provided in
products and the impact of laundering and sterilization on
Test Method F903. Diagrams of the test cell and apparatus are
reusable products. The integrity of the protective barrier may
provided in Figs. 1 and 2, respectively.
also be compromised during use by such effects as flexing and
6.1.3 Retaining Screen, a smooth finish plastic or metal
abrasion (8). Prewetting agents, such as alcohol, and contami-
square mesh screen meeting the following specifications is
nating agents, such as perspiration, may also compromise the
used for Procedure B from Table 1.
integrity of the protective barrier. If these conditions are of
concern, the performance of protective clothing materials
should be evaluated for Phi-X174 Bacteriophage penetration
TheU.S.FoodandDrugAdministration(FDA)hasspecificAcceptableQuality
Level (AQL) requirements for the use of this test method. Manufacturers,
distributors, and repackagers of surgical apparel should contact the FDA’s Division
of Small Manufacturers Assistance (DSMA).
TABLE 1 Specimen Exposure Procedures Thickness may be determined prior to performing the test procedure but is not
required. The thickness data for the material may be available from the manufac-
Procedure Pressure/Time Sequence and Retaining Screen Options
turer. Manipulation or handling of the test specimens prior to testing increases the
A 0 kPa [0 psig] for 5 min, followed by 13.8 kPa [2 psig]
chance for contamination.
for 1 min, followed by 0 kPa [0 psig] for 54 min.
The sole source of supply of the penetration test apparatus known to the
A retaining screen is not used to support the specimen.
committeeatthistimeisWilsonRoadMachineShop,1170WilsonRd.,RisingSun,
MD 21911.
B 0 kPa [0 psig] for 5 min, followed by 13.8 kPa [2 psig]
This is the sole source of supply of the apparatus known to the committee at
for 1 min, followed by 0 kPa [0 psig] for 54 min.
this time. If you are aware of alternative suppliers, please provide this information
A retaining screen is used to support the specimen. The
type must be specified. to ASTM Headquarters. Your comments will receive careful consideration at a
meeting of the responsible technical committee, which you may attend.
F1671/F1671M − 22
6.2.1 Petri Dishes, sterile, 15 by 100 mm.
6.2.2 Pipettes, sterile, 1, 5, and 10 mL.
6.2.3 Test Tubes, 13 by 100 mm.
6.2.4 Test Tube Rack, stainless.
11,13
6.2.5 Membrane Filters, sterile, 0.22 µm.
6.2.6 Bottles, sterile, glass, 100 to 500mL capacity.
6.2.7 Micropipets, capable of delivery of 2 µL, accurately
and consistently.
7. Reagents
14,15,11
7.1 Bacteriophage, Phi-X174, ATCC 13706-B1.
NOTE 1—The surrogate microbe, Phi-X174 Bacteriophage, was se-
lected as the most appropriate model for blood-borne pathogens because
of its small size, spherical (icosahedral) morphology, environmental
stability, nonhuman infectivity, high-assay sensitivity, rapid assay, and
high titer.The Phi-X174 Bacteriophage has no envelope and is one of the
smallest known viruses (0.027 µm in diameter). The Phi-X174 Bacterio-
phage is used at a challenge titer of at least 1.0×10 PFU/mL (plaque
forming units per millilitre).
14,11
7.2 Bacteria, E. coli C., ATCC 13706.
7.3 Purified Water, Q.S.
16,11
7.4 Nutrient Broth.
7.5 Calcium Chloride (CaCl ).
7.6 Potassium Chloride (KCl).
7.7 Sodium Hydroxide (NaOH), N solution.
17,11
FIG. 1 Exploded View of the Penetration Test Cell with Retaining
7.8 Surfactant, polysorbate 80.
Screen
16,11
7.9 Bacto-Agar.
8. Hazards
% open area >50
Limits deflection of specimen to 8.1 Sterilize all apparatus and supplies which come into
Can be sterilized by either ethylene oxide or autoclaving.
Material of construction meets criteria in Section 13. contact with the Phi-X Bacteriophage, before and after con-
ducting this test method, by autoclaving from 121 to 123°C
6.1.4 Air Pressure Source, capable of providing air at 13.8
and from 207 to 221 kPa absolute [30 to 32 psia] for 15 min.
6 1.38 kPa [2.0 6 0.2 psig].
Othermethodsofsterilizationmaybeusedprovidedthereisno
6.1.5 Incubator, capable of maintaining 35 to 37°C.
inhibitionofthetestorganismsduringthesubsequentcourseof
6.1.6 Water Bath, capable of maintaining 45 6 2°C.
the test. Exercise extreme care to avoid contamination of the
6.1.7 Analytical Balance, capable of weighing 0.001 g and
laboratory spaces by complete sterilization or high level
suitable for measuring weight to the nearest 10 g/m [0.1
disinfection of all apparatus and supplies. This will reduce the
oz/yd ] in accordance with Test Method D3776/D3776M.
possibility of laboratory contamination.
(Optional.)
8.1.1 Iftestspecimensaresterilized,ensurethatthemethod
6.1.8 Vortex Mixer.
of sterilization chosen does not unintentionally affect the
6.1.9 Refrigerator, capable of maintaining 2 to 8°C.
performance of the test specimen. Sterilize test specimens in
6.1.10 Autoclave,capableofmaintaining121to123°Cand
accordance with manufacturer’s recommendations.
207 to 221 kPa absolute [30 to 32 psia].
6.1.11 Stopwatch or Electronic Timer.
8.2 Although there is no known hazard for human infection
6.1.12 Orbital Shaker.
from Phi-X174 Bacteriophage, test operators should avoid
6.1.13 pH Meter, sensitive to 0.1 pH units.
direct contact with liquids containing the bacteriophage.
6.1.14 Inoculating Loop.
6.1.15 Torque Wrench, capable of 13.6 N·m [120 in.-lb].
6.1.16 Spectrophotometer, capable of 640 nm. 13
Membrane filters shall be used which meet requirements of Test Method
D3862.
6.1.17 Centrifuge, capable of 10000×g.
Cultures may be obtained from ATCC American Type Culture Collection,
6.2 Materials:
12301 Parklawn Drive, Rockville, MD 20852.
PurifiedviruspreparationsmaybeobtainedfromPromegaCorp.,2800Woods
Hollow Rd., Madison, WI 53711-5399.
12 16
Theweightmaybedeterminedpriortoperformingthetestprocedurebutisnot Nutrient Broth and Bacto-Agar may be obtained from Difco, Detroit, MI
required. The basis weight of the material may be available from the manufacturer. 48232.
Manipulation or handling of the test specimens prior to testing increases the chance Surfactant type Polysorbate 80 is suitable for this use and may be obtained
for contamination. fromAldrichChemicalCo.,Inc.,1001WestSaintParkAve.,Milwaukee,WI53233.
F1671/F1671M − 22
FIG. 2 Three-Dimensional Side View of the Test Apparatus
8.3 Either keep a transparent safety shield between the 9.1.4 Sterilize the bacteriophage nutrient broth in the auto-
penetration cell and the observer or, alternatively, perform the clave.
tests behind the window in a safety hood.
9.1.5 Measure the resulting surface tension of the sterile
solution using Test Method D1331. Do not use the bacterio-
9. Prepa
...

This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: F1671/F1671M − 13 F1671/F1671M − 22
Standard Test Method for
Resistance of Materials Used in Protective Clothing to
Penetration by Blood-Borne Pathogens Using Phi-X174
Bacteriophage Penetration as a Test System
This standard is issued under the fixed designation F1671/F1671M; 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 (´) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
Workers, primarily those in the health care profession, involved in treating and caring for
individuals injured or sick, can be exposed to biological liquids capable of transmitting disease. These
diseases, which may be caused by a variety of microorganisms, can pose significant risks to life and
health. This is especially true of blood-borne viruses which cause Hepatitis (Hepatitis B Virus (HBV)
and Hepatitis C Virus (HCV)) and Acquired Immune Deficiency Syndrome (AIDS) (Human
Immunodeficiency Virus (HIV)). Since engineering controls can not eliminate all possible exposures,
attention is placed on reducing the potential of direct skin contact through the use of protective
clothing that resists penetration (29 CFR Part 1910.1030). This test method was developed to assess
the effectiveness of materials used in protective clothing for protecting the wearer against contact with
blood-borne pathogens using a surrogate microbe suspended in a body fluid simulant under conditions
of continuous contact.
1. Scope
1.1 This test method is used to measure the resistance of materials used in protective clothing to penetration by blood-borne
pathogens using a surrogate microbe under conditions of continuous liquid contact. Protective clothing material pass/fail
determinations are based on the detection of viral penetration.
1.1.1 This test method is not always effective in testing protective clothing materials having thick, inner liners which readily
absorb the liquid assay fluid.
1.2 This test method does not apply to all forms or conditions of blood-borne pathogen exposure. Users of the test method should
review modes for worker/clothing exposure and assess the appropriateness of this test method for their specific applications.
1.3 This test method has been specifically defined for modeling the viral penetration of Hepatitis (B and C) and Human
Immunodeficiency Viruses transmitted in blood and other potentially infectious body fluids. Inferences for protection from other
pathogens must be assessed on a case-by-case basis.
1.4 This test method addresses only the performance of materials or certain material constructions (for example, seams) used in
This test method is under the jurisdiction of ASTM Committee F23 on Personal Protective Clothing and Equipment and is the direct responsibility of Subcommittee
F23.40 on Biological.
Current edition approved May 1, 2013Jan. 1, 2022. Published June 2013January 2022. Originally approved in 1995. Last previous edition approved in 20072013 as
F1671 – 07.F1671/F1671M – 13. DOI: 10.1520/F1671_F1671M-13.10.1520/F1671_F1671M-22.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F1671/F1671M − 22
protective clothing and determined to be viral resistant. This test method does not address the design, overall construction and
components, or interfaces of garments or other factors which may affect the overall protection offered by the protective clothing.
1.5 The values stated in SI units or in other units shall be regarded separately as standard. The values stated in each system must
be used independently of the other, without combining values in any way.
1.6 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 safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.7 This international standard was developed in accordance with internationally recognized principles on standardization
established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued
by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
2. Referenced Documents
2.1 ASTM Standards:
D1331 Test Methods for Surface and Interfacial Tension of Solutions of Paints, Solvents, Solutions of Surface-Active Agents,
and Related Materials
D1777 Test Method for Thickness of Textile Materials
D3776D3776/D3776M Test Methods for Mass Per Unit Area (Weight) of Fabric
D3862 Test Method for Retention Characteristics of 0.2-μm Membrane Filters Used in Routine Filtration Procedures for the
Evaluation of Microbiological Water Quality
E105 Guide for Probability Sampling of Materials
E171E171/E171M Practice for Conditioning and Testing Flexible Barrier Packaging
F903 Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Liquids
F1670F1670/F1670M Test Method for Resistance of Materials Used in Protective Clothing to Penetration by Synthetic Blood
2.2 Military Standard:
MIL-STD-105 Sampling Procedures and Tables for Inspection by Attributes
2.3 ANSI/ASQCANSI/ASQ Standard:
ANSI/ASQCANSI/ASQ Z1.4 Sampling Procedures and Tables for Inspection by Attributes
2.4 ISO Standard:
ISO 2859-1 Sampling Plans for Inspection by Attributes
2.5 OSHA Standard:
29 CFR Part 1910.1030 “Occupational Exposure to Bloodborne Pathogens: Final Rule,” Bloodborne PathogensFederal
Register, Vol 56, No. 235, Dec. 6, 1991, pp. 64175-64182.
3. Terminology
3.1 Definitions:
3.1.1 agar, n—a semisolid culture medium used to support the growth of bacteria and other micro-organisms.microorganisms.
3.1.2 aseptic, adj—sterile, free from viable microbiological contamination.
3.1.3 assay, n—analysis of a mixture to determine the presence or concentration of a particular component.
3.1.3.1 Discussion—
In this test method, the component being analyzed is a microorganism, Phi-X174 Bacteriophage.
3.1.4 assay fluid, n—a sterile liquid used to wash the test specimen surface to determine microbiological penetration.
3.1.4.1 Discussion—
In this test method, the assay fluid is bacteriophage nutrient broth and the microorganism is the Phi-X174 Bacteriophage. The assay
fluid is used to wash the Phi-X174 Bacteriophage from the normal inside surface of the test specimen.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
Available from Standardization Documents Order Desk, DODSSP, Bldg. 4, Section D, 700 Robbins Ave., Philadelphia, PA 19111-5098, http://dodssp.daps.dla.mil.
Available from American Society for Quality (ASQ), 600 N. Plankinton Ave., Milwaukee, WI 53203, http://www.asq.org.
Available from American National Standards Institute (ANSI), 25 W. 43rd St., 4th Floor, New York, NY 10036, http://www.ansi.org.
Available from U.S. Government Printing Office Superintendent of Documents, 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
www.access.gpo.gov.
F1671/F1671M − 22
3.1.5 bacteriophage, n—a type of virus which infects bacteria.
3.1.5.1 Discussion—
In this test method, the bacteriophage is Phi-X174. The Phi-X174 Bacteriophage is not pathogenic to humans, but serves to
simulate viruses that are pathogenic to humans.
3.1.6 blood-borne pathogen, n—an infectious bacterium or virus, or other disease inducing disease-inducing microbe carried in
blood or other potentially infectious body fluids.
3.1.6.1 Discussion—
For the purpose of this test method, the primary blood-borne pathogens include Hepatitis B Virus (HBV), Hepatitis C Virus (HCV),
and Human Immunodeficiency Virus (HIV). Other microorganisms must be considered on a case-by-case basis.
3.1.7 body fluid, n—any liquid produced, secreted, or excreted by the human body.
3.1.7.1 Discussion—
In this test method, body fluids include those liquids potentially infected with blood-borne pathogens, including, but not limited
to,to: blood, semen, vaginal secretions, cerebrospinal fluid, synovial fluid, peritoneal fluid, amniotic fluid, saliva in dental
procedures, any body fluid that is visibly contaminated with blood, and all body fluids in situations where it is difficult or
impossible to differentiate between body fluids (see section 29 CFR Part 1910.1030).
3.1.8 body fluid simulant, n—a liquid which is used to act as a model for human body liquids.
3.1.8.1 Discussion—
In this test method, the body fluid simulant is bacteriophage nutrient broth, which is intended as a model for human body liquids
as it approximates the lower end of the surface tension range for blood and body fluids (excluding saliva), 0.042 6 0.002 N/m.
3.1.9 challenge suspension, n—a liquid containing an agent that is used to test the penetration resistance of materials.
3.1.9.1 Discussion—
In this test method, the challenge suspension is the bacteriophage challenge suspension;suspension, a nutrient broth containing the
Phi-X174 Bacteriophage.
3.1.10 lawn, n—as in microbiology, a cloudy, uniform growth of bacteria in a thin layer of top agar in a petri dish.
3.1.10.1 Discussion—
In this test method, E. coli C. has been selected as the bacterium used to produce the lawn.
3.1.11 lysis, n—the disintegration or destruction of whole bacterial cells.
3.1.11.1 Discussion—
In this test method, the lysis of the host bacteria, E. coli C., is caused by Phi-X174 Bacteriophage.
3.1.12 medium (plural, media), n—a nutrient system for the cultivation of cells or organisms, and especially bacteria.
3.1.12.1 Discussion—
In this test method, the term media is used to describe mixtures compounded to support the growth of specific microorganisms;
for example, bacteriophage nutrient broth, top agar.
3.1.13 morphology, n—the form and structure of a particular organism.
3.1.14 nutrient broth, n—a liquid medium.
3.1.14.1 Discussion—
In this test method, the nutrient broth is the bacteriophage nutrient broth which is used to culture the host bacteria, E. coli C., and
to aid in manipulating the Phi-X174 Bacteriophage through the various stages of the procedure, such as;as suspending the
Phi-X174 Bacteriophage for challenging the test material in the penetration cell, assaying the normal inside test material surface
and, if required, making dilutions of the assay fluid for plating.
3.1.15 penetration, n—the movement of matter through closures, porous materials, seams, and pinholes or other imperfections in
protective clothing on a nonmolecular level.
F1671/F1671M − 22
3.1.15.1 Discussion—
For this test method, the specific matter is a bacteriophage challenge suspension containing nutrient broth with Phi-X174
Bacteriophage.
3.1.16 plaque, n—as in virology, a visible, clear area, which is theoretically the result of the infection and lysis of host cells by
a single viable virus.
3.1.16.1 Discussion—
In this test method, the term plaque is used to describe a visible, clear area,area in the lawn of E. coli C. in top agar, which is
theoretically the result of a single viable Phi-X174, where the bacteria have been destroyed by bacteriophage infection and lysis.
3.1.17 plaque forming unit (PFU), n—a virus particle capable of producing plaques by infecting and lysing bacteria in a lawn in
top agar.
3.1.18 plate, n—as in microbiology, a Petripetri dish containing culture medium.
3.1.19 protective clothing, n—an item of clothing that is specifically designed and constructed for the intended purpose of isolating
all or part of the body from a potential hazard; or, isolating the external environment from contamination by the wearer of the
clothing.
3.1.19.1 Discussion—
In this test method, materials used in protective clothing are being evaluated. The potential hazard of contact with blood-borne
pathogens is being simulated.
3.1.20 sterile, adj—free from viable microorganisms.
3.1.21 surrogate microbe, n—a microorganism which is used to act as a simulant for other microorganisms which are pathogenic
to humans.
3.1.21.1 Discussion—
In this test method, the surrogate microbe is the Phi-X174 Bacteriophage, intended as a model for HCV and to simulate both HBV
and HIV.
3.1.22 titer, n—the quantity of a substance required to react with, or to correspond to, a given amount of another substance.
3.1.22.1 Discussion—
In this test method, titer is used to describe the concentration of viable bacteriophage as measured in plaque forming units per
millilitre (PFU/mL).
3.1.23 viral penetration, n—the penetration of a material by a virus.
3.1.23.1 Discussion—
In this test method, viral penetration is used to describe the physical translocation of the Phi-X174 Bacteriophage through closures,
seams, pores, and pinholes or other imperfections in materials used in protective clothing.
3.1.24 viral resistant, adj—referring to materials which impede viral penetration under specified laboratory test conditions and
detection methods.
3.1.24.1 Discussion—
In this test method, protective clothing materials which demonstrate pass“pass” results are considered to be resistant to viral
penetration.
3.1.25 virus, n—a minute infectious agent,agent which lacks independent metabolism and is only able to replicate within a living
host cell.
4. Summary of Test Method
4.1 A specimen is subjected to a nutrient broth containing a virus for a specified time and pressure sequence.
4.2 Visual detection of penetration is supplemented with an assay procedure that will detect viable viruses which penetrate the
material even when liquid penetration is not visible. Any evidence of viral penetration for a test specimen constitutes failure.
F1671/F1671M − 22
5. Significance and Use
5.1 This test method is based on Test Method F903 for measuring resistance of chemical protective clothing materials to
penetration by liquids. This test method is normally used to evaluate specimens from individual finished items of protective
clothing and individual samples of materials that are candidates for items of protective clothing.
5.1.1 Finished items of protective clothing include gloves, arm shields, aprons, gowns, coveralls, hoods, and boots.
5.1.2 The phrase “specimens from finished items” encompasses seamed and other discontinuous regions, as well as the usual
continuous regions of protective clothing items.
5.2 It is known that body fluids penetrating protective clothing materials are likely to carry microbiological contaminants;
however, visual detection methods are not sensitive enough to detect minute amounts of liquid containing microorganisms (11-3,
2, 3). This test method uses media containing Phi-X174 Bacteriophage. The visual detection technique of this test method is
supplemented with a biologically based assay capable of detecting virus under the specified test conditions.
5.3 Test Method F1670F1670/F1670M, allows the screening of protective clothing materials for resistance to penetration with
synthetic blood as a challenge liquid. Test Method F1670F1670/F1670M uses the same penetration test cell and technique, but
exposes material specimens to synthetic blood with visual detection of liquid penetration. Materials passing Test Method
F1670F1670/F1670M should then be tested against bacteriophage penetration using this test method to verify performance.
5.4 This test method has been specifically designed for measuring penetration of a surrogate microbe for Hepatitis (B and C) and
the Human Immunodeficiency Viruses. The surrogate, Phi-X174 Bacteriophage, used in this test method is similar to HCV in size
and shape but also serves as a surrogate for HBV and HIV. Inferences about protection from other pathogens must be assessed on
a case-by-case basis.
5.5 Part of the protocol in ProcedureProcedures A and B in Table 1, for exposing the protective clothing material specimens to
the Phi-X174 Bacteriophage challenge suspension,suspension involves pressurization of the penetration cell to 13.8 kPa [2 psig].
This hydrostatic pressure has been documented to discriminate between protective clothing material performance and correlate
with visual penetration results that are obtained with a human factors validation (4). Some studies, however, suggest that
mechanical pressures exceeding 345 kPa [50 psig] can occur during actual clinical use (5, 6). Therefore, it is important to
understand that this test method does not simulate all the physical stresses and pressures that might be exerted on protective
clothing materials during actual use.
5.6 Medical protective clothing materials are intended to be a barrier to blood, body fluids, and other potentially infectious
materials. Many factors can effectaffect the wetting and penetration characteristics of body fluids, such as:as surface tension;
TABLE 1 Specimen Exposure Procedures
Procedure Pressure/Time Sequence and Retaining Screen Options
A 0 kPa [0 psig] for 5 min, followed by 13.8 kPa [2 psig]
for 1 min, followed by 0 kPa [0 psig] for 54 min.
A retaining screen is not used to support the specimen.
B 0 kPa [0 psig] for 5 min, followed by 13.8 kPa [2 psig]
for 1 min, followed by 0 kPa [0 psig] for 54 min.
A retaining screen is used to support the specimen. The
type must be specified.
TABLE 1 Specimen Exposure Procedures
Procedure Pressure/Time Sequence and Retaining Screen Options
A 0 kPa [0 psig] for 5 min, followed by 13.8 kPa [2 psig]
for 1 min, followed by 0 kPa [0 psig] for 54 min.
A retaining screen is not used to support the specimen.
B 0 kPa [0 psig] for 5 min, followed by 13.8 kPa [2 psig]
for 1 min, followed by 0 kPa [0 psig] for 54 min.
A retaining screen is used to support the specimen. The
type must be specified.
The boldface numbers in parentheses refer to the list of references at the end of this standard.
F1671/F1671M − 22
viscosity;tension, viscosity, and polarity of the fluids, as well as the structure and relative hydrophilicity or hydrophobicity of the
materials. The surface tension range for blood and body fluids (excluding saliva) is approximately 0.042 to 0.060 N/m (7). To help
simulate the wetting characteristics of blood and body fluids, the surface tension of the Phi-X174 Bacteriophage challenge
suspension is adjusted to approximate the lower end of this surface tension range. This is accomplished by adding surfactant to
the Phi-X174 Bacteriophage nutrient broth. The resulting surface tension of the Phi-X174 Bacteriophage challenge suspension is
approximately 0.042 6 0.002 N/m.
5.7 Testing prior to degradation by physical, chemical, and thermal stresses which could negatively impact the performance of the
protective material,material could lead to a false sense of security. Additional tests should be considered that assess the impact of
storage conditions and shelf life on disposable products and the impact of laundering and sterilization on reusable products. The
integrity of the protective barrier may also be compromised during use by such effects as flexing and abrasion (8). Prewetting
agents, such as alcohol, and contaminating agents, such as perspiration, may also compromise the integrity of the protective barrier.
If these conditions are of concern, the performance of protective clothing materials should be evaluated for Phi-X174
Bacteriophage penetration following an appropriate preconditioning technique representative of the expected conditions of use.
5.8 This test method involves a sensitive assay procedure for determining protective clothing material resistance to penetration by
a surrogate microbe. Because of the length of time required to complete this method, it may not be suitable for use as a material
or protective clothing quality control or quality assurance procedure.
5.9 If this procedure is used for quality control or to support broad product claims concerning the viral resistant viral-resistant
properties of materials used in protective clothing, proper statistical design and analysis of larger data sets than those specified in
this test method should be performed. Examples of acceptable sampling plans can be found in MIL-STD-105, ANSI/ASQCANSI/
ASQ Z1.4, and ISO 2859-1.
5.10 This test method requires a working knowledge of basic microbiological techniques (9).
6. Apparatus and Materials
6.1 Apparatus:
6.1.1 Thickness Gauge, suitable for measuring thickness to the nearest 0.02 mm [or nearest 0.001 in.], as specified in Test Method
D1777, to determine the thickness of each protective clothing material specimen tested. (Optional.)
10,11
6.1.2 Penetration Test Cell, to restrain the test specimen during contact with the pressurized Phi-X174 Bacteriophage
challenge suspension. In the test cell, the specimen acts as a partition separating the bacteriophage challenge suspension from the
viewing side of the penetration cell. The capacity of the cell body is approximately 60 mL [2.0 oz] for the challenge suspension.
A flange cover, with an open area to allow visual observation and assay of the specimen, and a transparent cover are included. The
cell body has a top port for filling and a drain valve for draining the penetration test cell. Other items, such as a fitting to allow
attachment of the air line to the top port in the cell body, gaskets, and the retaining screen will also be required. Specifications for
the penetration cell are provided in Test Method F903. Diagrams of the test cell and apparatus are provided in Figs. 1 and 2,
respectively.
6.1.3 Retaining Screen, a smooth finish plastic or metal square mesh screen meeting the following specifications is used for
Procedure B from Table 1.
% open area >50
Limits deflection of specimen to Can be sterilized by either ethylene oxide or autoclaving
Can be sterilized by either ethylene oxide or autoclaving.
Material of construction meets criteria in 15.4.
Material of construction meets criteria in Section 13.
The U.S. Food and Drug Administration (FDA) has specific Acceptable Quality Level (AQL) requirements for the use of this test method. Manufacturers, distributors,
and repackagers of surgical apparel should contact the FDA’s Division of Small Manufacturers Assistance (DSMA).
Thickness may be determined prior to performing the test procedure but is not required. The thickness data for the material may be available from the manufacturer.
Manipulation or handling of the test specimens prior to testing increases the chance for contamination.
The sole source of supply of the penetration test apparatus known to the committee at this time is Wilson Road Machine Shop, 1170 Wilson Rd., Rising Sun, MD 21911.
This is the sole source of supply of the apparatus known to the committee at this time. If you are aware of alternative suppliers, please provide this information to ASTM
Headquarters. Your comments will receive careful consideration at a meeting of the responsible technical committee, which you may attend.
F1671/F1671M − 22
FIG. 1 ExplodedExploded View of the Penetration Test Cell with Retaining Screen
6.1.4 Air Pressure Source, capable of providing air at 13.8 6 1.38 kPa [2.0 6 0.2 psig].
6.1.5 Incubator, capable of maintaining 35 to 37°C.37 °C.
6.1.6 Water Bath, capable of maintaining 45 6 2°C.2 °C.
2 2
6.1.7 Analytical Balance, capable of weighing 0.001 g and suitable for measuring weight to the nearest 10 g/m [0.1 oz/yd ] in
accordance with Test Method D3776D3776/D3776M. (Optional.)
6.1.8 Vortex Mixer.
6.1.9 Refrigerator, capable of maintaining 2 to 8°C.8 °C.
6.1.10 Autoclave, capable of maintaining 121 to 123°C123 °C and 207 to 221 kPa absolute [30 to 32 psia].
6.1.11 Stopwatch or Electronic Timer.
6.1.12 Orbital Shaker.
6.1.13 pH Meter, sensitive to 0.1 pH units.
6.1.14 Inoculating Loop.
6.1.15 Torque Wrench, capable of 13.6 N·m [120 in.-lb].
The weight may be determined prior to performing the test procedure but is not required. The basis weight of the material may be available from the manufacturer.
Manipulation or handling of the test specimens prior to testing increaseincreases the chance for contamination.
F1671/F1671M − 22
FIG. 2 Three Dimensional Three-Dimensional Side View of the Test Apparatus
6.1.16 Spectrophotometer, capable of 640 nm.
6.1.17 Centrifuge, capable of 10 000 × g.
6.2 Materials:
6.2.1 Petri Dishes, sterile sterile, 15 by 100 mm.
6.2.2 Pipettes, sterile, 1, 5, and 10 mL.
6.2.3 Test Tubes, 13 by 100 mm.
6.2.4 Test Tube Rack, stainless.
11,13
6.2.5 Membrane Filters, sterile, 0.22 μm.
6.2.6 Bottles, sterile, glass, 100 to 500-mL500 mL capacity.
6.2.7 Micropipets, capable of delivery of 2 μL, accurately and consistently.
Membrane filters shall be used which meet requirements of Test Method D3862.
F1671/F1671M − 22
7. Reagents
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7.1 Bacteriophage, Phi-X174, ATCC 13706-B1.
NOTE 1—The surrogate microbe, Phi-X174 Bacteriophage, was selected as the most appropriate model for blood-borne pathogens because of its small
size, spherical (icosahedral) morphology, environmental stability, nonhuman infectivity, high-assay sensitivity, rapid assay, and high titer. The Phi-X174
Bacteriophage has no envelope and is one of the smallest known viruses (0.027 μm in diameter). The Phi-X174 Bacteriophage is used at a challenge titer
of at least 1.0 × 10 PFU/mL (plaque forming units per millilitre).
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7.2 Bacteria, E. coli C., ATCC 13706.
7.3 Purified Water, Q.S.
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7.4 Nutrient Broth.
7.5 Calcium Chloride (CaCl ).
7.6 Potassium Chloride (KCl).
7.7 Sodium Hydroxide (NaOH), N solution.
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7.8 Surfactant, polysorbate 80.
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7.9 Bacto-Agar.
8. Hazards
8.1 Sterilize all apparatus and supplies which come into contact with the Phi-X Bacteriophage, before and after conducting this
test method, by autoclaving from 121 to 123°C123 °C and from 207 to 221 kPa absolute [30 to 32 psia] for 15 min. Other methods
of sterilization may be used provided there is no inhibition of the test organisms during the subsequent course of the test. Exercise
extreme care to avoid contamination of the laboratory spaces by complete sterilization or high level disinfection of all apparatus
and supplies. This will reduce the possibility of laboratory contamination.
8.1.1 If test specimens are sterilized, ensure that the method of sterilization chosen does not unintentionally affect the performance
of the test specimen. Sterilize test specimens in accordance with manufacturer’s recommendations.
8.2 Although there is no known hazard for human infection from Phi-X174 Bacteriophage, test operators should avoid direct
contact with liquids containing the bacteriophage.
8.3 Either keep a transparent safety shield between the penetration cell and the observer or, alternatively, perform the tests behind
the window in a safety hood.
9. Preparation of Media
9.1 Bacteriophage Nutrient Broth (Phi-X):
9.1.1 Prepare bacteriophage nutrient broth using the following:
Cultures may be obtained from ATCC American Type Culture Collection, 12301 Parklawn Drive, Rockville, MD 20852.
Purified virus preparations may be obtained from Promega Corp., 2800 Woods Hollow Rd., Madison, WI 53711-5399.
Nutrient Broth and Bacto-Agar may be obtained from Difco, Detroit, MI 48232.
Surfactant type Polysorbate 80 is suitable for this use and may be obtained from Aldrich Chemical Co., Inc., 1001 West Saint Park Ave., Milwaukee, WI 53233.
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Nutrient broth 8.0 ± 0.1 g
Potassium chloride 5.0 ± 0.06 g
Calcium chloride 0.2 ± 0.003 g
Purified water, Q.S. to 1000 ± 12.5 mL
Surfactant 0.1 ± 0.00125 mL (9.1.3)
9.1.2 Adjust the pH of the bacteriophage nutrient broth to 7.2 – 7.47.2 to 7.4 using 2.5 N sodium hydroxide.
9.1.3 Dilute 1one volume of 0.1 % surfactant with nine volumes of bacteriophage nutrient broth. To ensure adequate mixing, prior
to sterilization, heat the bacteriophage nutrient broth while stirring in the surfactant. A final concentration of 0.01 % surfactant is
recommended to adjust the final surface tension to 0.042 6 0.002 N/m.
9.1.4 Sterilize the bacteriophage nutrient broth in the autoclave.
9.1.5 Measure the resulting surface tension of the sterile solution using Test Method D1331. Do not use the bacteriophage nutrient
broth unless the corrected surface tension is within the 0.042 6 0.002-N/m 0.002 N/m range.
9.2 Bottom Agar (Phi-X):
9.2.1 Prepare bottom agar using the following:
Bacto-agar 15.0 ± 0.19 g
Nutrient broth 8.0 ± 0.1 g
Potassium chloride 5.0 ± 0.06 g
Purified water, Q.S. to 1000 ± 12.5 mL
Calcium chloride 1.0 ± 0.0125 mL
(Add sterile calcium chloride after autoclaving the bottom agar, 9.2.2.)
9.2.2 Prepare sterile calcium chloride by autoclaving a 1-
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