ASTM E3135-18

Standard Practice for Determining Antimicrobial Efficacy of Ultraviolet Germicidal Irradiation Against Microorganisms on Carriers with Simulated Soil

SIGNIFICANCE AND USE
5.1 This practice determines the effectiveness of UVGI devices for reducing viable microorganisms deposited on carriers.  
5.2 This practice evaluates the effect soiling agents have on UVGI antimicrobial effectiveness.  
5.3 This practice determines the delivered UVGI dose.
SCOPE
1.1 This practice will define test conditions to evaluate ultraviolet germicidal irradiation (UVGI) light devices (mercury vapor bulbs, light-emitting diodes, or xenon arc lamps) that are designed to kill/inactivate microorganisms deposited on inanimate carriers.  
1.2 This practice defines the terminology and methodology associated with the ultraviolet (UV) spectrum and evaluating UVGI dose.  
1.3 This practice defines the testing considerations that can reduce UVGI surface kill effectiveness, that is, presence of a soiling agent.  
1.4 This practice does not address shadowing.  
1.5 This practice should only be used by those trained in microbiology and in accordance with the guidance provided by Biosafety in Microbiological and Biomedical Laboratories (5th edition), 2009, HHS Publication No. (CDC) 21-1112.  
1.6 This practice does not recommend either specific test microbes or growth media. Users of this practice shall select appropriate test microbes and growth media based on the specific objectives of their UV antimicrobial performance evaluation test plan.  
1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.8 Warning—Mercury has been designated by many regulatory agencies as a hazardous substance that can cause serious medical issues. Mercury, or its vapor, has been demonstrated to be hazardous to health and corrosive to materials. Caution should be taken when handling mercury and mercury-containing products. See the applicable product Safety Data Sheet (SDS) for additional information. Users should be aware that selling mercury or mercury-containing products, or both, may be prohibited by local or national law.  
1.9 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.  
1.10 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.

General Information

• Status: Published
• Publication Date: 30-Apr-2018
• ICS: 07.100.01 - Microbiology in general
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents
• Drafting Committee: E35.15 - Antimicrobial Agents

Relations

• Refers: ASTM E1316-24 - Standard Terminology for <?Pub Dtl?>Nondestructive Examinations
• Effective Date: 01-Feb-2024
• Refers: ASTM G130-12(2020) - Standard Test Method for Calibration of Narrow- and Broad-Band Ultraviolet Radiometers Using a Spectroradiometer
• Effective Date: 01-Jun-2020
• Refers: ASTM E1053-20 - Standard Practice to Assess Virucidal Activity of Chemicals Intended for Disinfection of Inanimate, Nonporous Environmental Surfaces
• Effective Date: 01-Jan-2020
• Refers: ASTM E1316-19b - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Dec-2019
• Refers: ASTM E2756-19 - Standard Terminology Relating to Antimicrobial and Antiviral Agents
• Effective Date: 01-Nov-2019
• Refers: ASTM E1316-19 - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Mar-2019
• Refers: ASTM E2756-18 - Standard Terminology Relating to Antimicrobial and Antiviral Agents
• Effective Date: 01-Apr-2018
• Refers: ASTM E1316-18 - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Jan-2018
• Refers: ASTM E1316-17a - Standard Terminology for Nondestructive Examinations
• Effective Date: 15-Jun-2017
• Refers: ASTM E1316-17 - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Feb-2017
• Refers: ASTM E1316-16a - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Aug-2016
• Refers: ASTM E1316-16 - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Feb-2016
• Refers: ASTM E1316-15a - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Dec-2015
• Refers: ASTM E1316-15 - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Sep-2015
• Refers: ASTM E1316-14 - Standard Terminology for Nondestructive Examinations
• Effective Date: 01-Jun-2014

Overview

ASTM E3135-18 is a standard practice established by ASTM International for determining the antimicrobial efficacy of ultraviolet germicidal irradiation (UVGI) against microorganisms deposited on carriers with simulated soil. This practice is designed to assess how UVGI light sources, such as mercury vapor bulbs, light-emitting diodes (LEDs), and xenon arc lamps, perform in reducing viable microorganisms from non-living surfaces. It defines the required test conditions, relevant terminology, testing methodologies, and evaluation of UVGI dose, with a particular focus on the influence of soiling agents-substances that can reduce the effectiveness of UV disinfection.

This standard is essential for laboratories, manufacturers, and regulators involved in designing, using, or certifying UVGI devices for surface decontamination, especially in healthcare, laboratory, and public health contexts.

Key Topics

• UVGI Device Evaluation
  The standard provides a framework to assess the performance of various UVGI devices intended to inactivate or kill microorganisms on inanimate carriers.

• Effect of Soiling Agents
  Evaluates how substances mimicking real-world contaminants (simulated soil) can affect the antimicrobial performance of UVGI. Both hydrophilic and hydrophobic soiling agents are considered.

• Dose Determination
  Details on how to measure and calculate the delivered dose of UVGI, taking into account irradiance, time, and placement to ensure consistent and reliable results.

• Microbial Carrier Testing
  Defines methods for contaminating carriers, applying soiling agents, exposing samples to UVGI, extracting and enumerating surviving microorganisms, and calculating percent reduction.

• Safety and Hazard Considerations
  General guidance on laboratory biosafety, safe handling of mercury-containing products, and managing the risks associated with UV-C radiation and potential ozone production.

Applications

Organizations and professionals use ASTM E3135-18 in the following practical scenarios:

• Validation of UVGI Devices
  Manufacturers and laboratories use this standard to validate and compare the antimicrobial effectiveness of different UVGI systems before market release or deployment.

• Development of Infection Control Protocols
  Hospitals and public health facilities rely on ASTM E3135-18 to determine UVGI device suitability for disinfection protocols, particularly in environments where surfaces may be soiled.

• Quality Assurance and Regulatory Compliance
  Regulatory authorities can reference this standard in certification programs or approval processes for UVGI equipment intended for environmental disinfection.

• Research and Development
  Scientists and engineers apply the standard’s methodology when investigating new materials, technologies, or configurations for ultraviolet surface disinfection.

• Performance Benchmarking under Realistic Conditions
  By including simulated soiling agents, the standard offers a more accurate prediction of UVGI device performance in real-world settings with typical surface contamination.

Related Standards

Several related standards provide complementary testing methodologies and definitions relevant to ASTM E3135-18, including:

• ASTM E1053: Test Method to Assess Virucidal Activity of Chemicals for Disinfection of Inanimate, Nonporous Environmental Surfaces
• ASTM E1153: Test Method for Efficacy of Sanitizers for Inanimate, Hard, Nonporous Non-Food Contact Surfaces
• ASTM E1316: Terminology for Nondestructive Examinations
• ASTM E2756: Terminology Relating to Antimicrobial and Antiviral Agents
• ASTM E2721: Practice for Evaluation of Effectiveness of Decontamination Procedures for Surfaces Challenged with Pathogenic Viruses
• ASTM G130: Test Method for Calibration of Ultraviolet Radiometers
• ISO 9370: Plastics - Instrumental Determination of Radiant Exposure in Weathering Tests - General Guidance

Compliance with ASTM E3135-18 supports best practices and aligns with international principles for standards development and regulatory acceptance of UVGI antimicrobial performance testing.

Keywords: UVGI, ultraviolet germicidal irradiation, antimicrobial efficacy, simulated soil, surface decontamination, ASTM E3135-18, UV-C, disinfection standards, infection control, laboratory testing.

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: E3135 − 18
Standard Practice for
Determining Antimicrobial Efficacy of Ultraviolet Germicidal
Irradiation Against Microorganisms on Carriers with
Simulated Soil
This standard is issued under the fixed designation E3135; the number immediately following the designation indicates the year 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.
1. Scope that selling mercury or mercury-containing products, or both,
may be prohibited by local or national law.
1.1 This practice will define test conditions to evaluate
1.9 This standard does not purport to address all of the
ultraviolet germicidal irradiation (UVGI) light devices (mer-
safety concerns, if any, associated with its use. It is the
cury vapor bulbs, light-emitting diodes, or xenon arc lamps)
responsibility of the user of this standard to establish appro-
that are designed to kill/inactivate microorganisms deposited
priate safety, health, and environmental practices and deter-
on inanimate carriers.
mine the applicability of regulatory limitations prior to use.
1.2 This practice defines the terminology and methodology
1.10 This international standard was developed in accor-
associated with the ultraviolet (UV) spectrum and evaluating
dance with internationally recognized principles on standard-
UVGI dose.
ization established in the Decision on Principles for the
1.3 This practice defines the testing considerations that can
Development of International Standards, Guides and Recom-
reduce UVGI surface kill effectiveness, that is, presence of a
mendations issued by the World Trade Organization Technical
soiling agent.
Barriers to Trade (TBT) Committee.
1.4 This practice does not address shadowing.
2. Referenced Documents
1.5 This practice should only be used by those trained in
2.1 ASTM Standards:
microbiologyandinaccordancewiththeguidanceprovidedby
E1053Test Method to Assess Virucidal Activity of Chemi-
BiosafetyinMicrobiologicalandBiomedicalLaboratories(5th
cals Intended for Disinfection of Inanimate, Nonporous
edition), 2009, HHS Publication No. (CDC) 21-1112.
Environmental Surfaces
1.6 This practice does not recommend either specific test
E1153TestMethodforEfficacyofSanitizersRecommended
microbes or growth media. Users of this practice shall select
for Inanimate, Hard, Nonporous Non-Food Contact Sur-
appropriate test microbes and growth media based on the
faces
specific objectives of their UV antimicrobial performance
E1316Terminology for Nondestructive Examinations
evaluation test plan.
E2756Terminology Relating toAntimicrobial andAntiviral
Agents
1.7 The values stated in SI units are to be regarded as
E2721Practice for Evaluation of Effectiveness of Decon-
standard. No other units of measurement are included in this
tamination Procedures for Surfaces When Challenged
standard.
with Droplets Containing Human Pathogenic Viruses
1.8 Warning—Mercuryhasbeendesignatedbymanyregu-
G130Test Method for Calibration of Narrow- and Broad-
latoryagenciesasahazardoussubstancethatcancauseserious
Band Ultraviolet Radiometers Using a Spectroradiometer
medicalissues.Mercury,oritsvapor,hasbeendemonstratedto
2.2 ISO Standards:
be hazardous to health and corrosive to materials. Caution
9370Plastics -- Instrumental Determination of Radiant Ex-
should be taken when handling mercury and mercury-
posureinWeatheringTests--GeneralGuidanceandBasic
containing products. See the applicable product Safety Data
Test Method
Sheet(SDS)foradditionalinformation.Usersshouldbeaware
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
This practice is under the jurisdiction ofASTM Committee E35 on Pesticides, Standards volume information, refer to the standard’s Document Summary page on
Antimicrobials, and Alternative Control Agents and is the direct responsibility of the ASTM website.
Subcommittee E35.15 on Antimicrobial Agents. Available from International Organization for Standardization (ISO), ISO
Current edition approved May 1, 2018. Published July 2018. DOI: 10.1520/ Central Secretariat, BIBC II, Chemin de Blandonnet 8, CP 401, 1214 Vernier,
E3135–18 Geneva, Switzerland, http://www.iso.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E3135 − 18
21348Space Environment (Natural andArtificial) – Process 4. Summary of Practice
for Determining Solar Irradiances
4.1 This practice describes the steps required to deposit
2.3 AOAC Methods: microorganisms onto carriers.
961.02Germicidal Spray Products as Disinfectants
4.2 This practice defines the process for adding soiling
agents on top of microorganisms, which can reduce the
3. Terminology
effectiveness of UV antimicrobial activity.
3.1 For Definitions of Terms used in this practice, refer to
4.3 This practice defines a protocol for quantifying the dose
Terminologies in E1316, E2756, and ISO 21348:
a UVGI device delivers to a surface.
3.2 Definitions:
4.4 This practice defines the process for exposure of micro-
3.2.1 carrier, n—a surrogate surface or matrix that facili-
organisms to UVGI.
tates the interaction of test microorganisms and treatment(s).
4.5 This practice defines protocols for extraction of viable
3.2.2 irradiance (E), n—a radiometric term for the radiant
microbes from carriers followed by enumeration.
flux that is incident upon a surface (W•m-2).
5. Significance and Use
3.2.3 joule (J), n—a unit of work or energy in the SI system
5.1 This practice determines the effectiveness of UVGI
of units.
devices for reducing viable microorganisms deposited on
3.2.3.1 Discussion—One Joule is one watt-second.
carriers.
3.2.4 light-emitting diode (LED), n—a solid-state electronic
5.2 This practice evaluates the effect soiling agents have on
device or transistor which emits light.
UVGI antimicrobial effectiveness.
3.2.4.1 Discussion—An LED is a p-n junction diode, which
emits light when activated. When a suitable voltage is applied
5.3 This practice determines the delivered UVGI dose.
totheleads,electronsareabletorecombinewithelectronholes
6. Hazards
within the device, releasing energy in the form of photons.
6.1 UV light becomes increasingly hazardous as the wave-
3.2.5 mercury vapor lamp, n—a gas discharge lamp that
length decreases, shifting from longer wavelengths (UV-A,
uses electric arc through vaporized mercury to produce light.
UV-B) to shorter wavelengths (UV-C). UV-A and UV-B are
3.2.6 radiometer, n—a device for measuring the radiant
part of the normal solar spectrum found in our atmosphere and
power that has an output proportional to the intensity of the
are responsible for UV-related aging, sunburns, and mutagenic
input power.
effects.UV-CradiationisfilteredoutbytheEarth’satmosphere
3.2.7 shadowing, v—creating a dark area or shape by
and is not part of the received solar spectrum. UV-C is highly
blocking light rays.
mutagenic and is harmful to all life forms.
3.2.8 soiling agent, n—substance applied either along with
NOTE 1—This practice is not designed to evaluate safety concerns
or on top of the test microorganism that can reduce the
surrounding UV exposure.
effectiveness of the antimicrobial technology.
6.2 Some UVGI bulbs produce ozone, which is harmful to
3.2.9 ultraviolet germicidal irradiation (UVGI), n—a
all life forms. Consult the manufacturer of the UVGI bulbs or
method that uses short-wavelength ultraviolet (UV-C) light to the device, or both, to determine if ozone is produced. If so,
killorinactivatemicroorganismsprimarilybyformingpyrimi-
you must follow Occupational Safety and Health Administra-
dine dimers, leaving them unable to perform vital cellular tion (OSHA) regulations to ensure worker safety.
functions.
NOTE 2—This practice is not designed to evaluate safety concerns
3.2.10 ultraviolet (UV) light, n—radiation having wave- surrounding ozone production by UVGI devices.
lengthsshorterthanwavelengthsofvisiblelight(~400nm)and
6.3 Safety measures are required to ensure workers are not
longer than those of X-rays (~100 nm).
exposed to UV light during testing, especially UV-B and
3.2.11 UV-A, n—radiation within the ultraviolet spectrum UV-C. Safety glasses with appropriate UV protection and
appropriate lab attire shall be used at all times when working
thatextendsfromapproximately315to400nminwavelength.
with UV devices.
3.2.12 UV-B, n—radiation within the ultraviolet spectrum
thatextendsfromapproximately280to315nminwavelength. 6.4 Signage shall be posted on the laboratory when UV
lights are in use to prevent accidental exposure to coworkers.
3.2.13 UV-C, n—radiation within the ultraviolet spectrum
which extends from approximately 100 to 280 nm in wave-
7. Reagents and Materials
length.
7.1 Microorganisms—selected by user based on the overall
3.2.14 xenon arc lamp, n—a specialized type of gas dis-
application.
charge lamp, an electric light that produces light by passing
electricity through ionized xenon gas at high pressure.
Reagent Chemicals, American Chemical Society Specifications, American
Chemical Society, Washington, DC. For Suggestions on the testing of reagents not
listed by the American Chemical Society, see Annual Standards for Laboratory
Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
Available from AOAC International, 2275 Research Blvd., Suite 300, and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
Rockville, MD 20850-3250, http://www.aoac.org. MD.
E3135 − 18
7.2 Soiling Agents—selected by user based on the overall 9.3.1 Radiometers are normally set to measure a band of
application. wavelengthssurroundingthepeakintensitywavelength,which
will vary among manufacturers.
7.3 50-mL Centrifuge Tubes—sterile, with caps.
9.3.2 The peak intensity for UV-C is 254 nm, and this
7.4 Growth Media and Supplemental Reagents—selectedby
wavelength should be used as the reference point for UV
user based on the overall application.
output from the UVGI light source and for measurement with
the radiometer.
7.5 Cell Spreaders—disposable, plastic triangular 60mm,
9.3.3 The irradiance is measured at the distance the carrier
sterile.
will be positioned away from the UVGI light source.
7.6 Petri Dishes—100 × 15 mm, sterile; glass or plastic.
9.3.3.1 The irradiance measured by most radiometers is
7.7 Carriers—2.5cmindiametercomposedofanymaterial. generally provided in power per square centimeter.
9.3.3.2 Irradiance is represented as a unit of watts (for
8. Equipment example, microwatt, milliwatt, etc.) resulting in a unit descrip-
tion of xW/cm .
8.1 UV Exposure Device—UVGI light source that is a
9.3.3.3 Delivered dose is a function of irradiance and time,
standalone bulb or contained within an exposure system.
2 2
which is expressed as J/cm (W/cm × Time (seconds of
8.2 Radiometer—calibrated to measure 254-nm irradiation
exposure)).
in accordance with Test Method G130 or ISO 9370.
9.3.3.4 Final dose should be expressed in J/cm .
8.3 Spectrophotometer—calibrated to measure wavelengths
NOTE 5—The intensity of UVGI will vary significantly over the length
ranging from at least 200 – 315 nm. Typically, a cosine ofthebulb,withthehighestirradianceb
...