IEC/PAS 63086-3-1:2023
(Main)Household and similar electrical air cleaning appliances — Methods for measuring the performance — Part 3-1: Method for assessing the reduction rate of key bioaerosols by portable air cleaners using an aerobiology test chamber
Household and similar electrical air cleaning appliances — Methods for measuring the performance — Part 3-1: Method for assessing the reduction rate of key bioaerosols by portable air cleaners using an aerobiology test chamber
This document specifies a method to evaluate the capability of portable household air cleaners to reduce the concentration and viability of key experimentally generated bioaerosols in a specified chamber. Indoor air free of harmful microbes is important to the health of occupants. This is particularly relevant with regard to increased time spent indoors. Air cleaners are used to reduce the concentrations of microorganisms in indoor air. The efficiency of such air cleaners to reduce airborne microorganisms can be assessed in test chambers at controlled air temperature and relative air humidity. The test is applicable to portable air cleaners commonly used in single room spaces such as those based on mechanical filtration, ultraviolet (UV), ionizers, photocatalytic oxidation, and ozone generators in-unit technology. If the air cleaner does not claim to have the function of reducing microorganisms, this document may not be applicable unless it is being used to simply evaluate the performance. This document deals with measurement procedures regarding the reduction of the microbial contamination related to electrical air cleaner appliances for household and similar use. This document does not apply to appliances intended to be used in medical, veterinary, or pharmaceutical applications. This document does not address sanitization, disinfection, or sterilization measures. This document does not support, by itself any health-related claims or conclusions about prevention or treatment of a disease or health improvement. NOTE 1 IEC 63086-3-1 is created for household and similar electrical air cleaners and is not intended to conflict with or replace standards for commercial or industrial consumers. NOTE 2 In this document, we do not suggest performance test methods that measure the by-products of either the interaction between microbes or between the air cleaner and the microbes tested in this document. The formation of by-products is an important subject. The subject of measuring by-products is under study, and AHAM will address this in future documents. NOTE 3 This document does not apply to appliances intended for use in medical treatment locations, such as surgical suites, laboratories, medical treatment rooms, etc.
Appareils électrodomestiques et analogues de purification de l'air — Méthodes de mesure de la performance — Partie 3-1: Titre manque
General Information
Standards Content (Sample)
FINAL
PUBLICLY IEC/DPAS
DRAFT
AVAILABLE 63086-3-1
SPECIFICATION
ISO/TC 142
Household and similar electrical air
Secretariat: UNI
cleaning appliances — Methods for
Voting begins on:
2022-12-21 measuring the performance —
Voting terminates on:
Part 3-1:
2023-02-15
Particular requirements for reduction
of microorganisms
Appareils électrodomestiques et analogues de purification de l'air —
Méthodes de mesure de la performance —
Partie 3-1: Exigences particulières pour la réduction des
microorganismes
This draft is submitted to a parallel vote in ISO and in IEC.
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THEY ARE AWARE AND TO PROVIDE SUPPOR TING
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BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO-
IEC/DPAS 63086-3-1:2022(E)
LOGICAL, COMMERCIAL AND USER PURPOSES,
DRAFT INTERNATIONAL STANDARDS MAY ON
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DARDS TO WHICH REFERENCE MAY BE MADE IN
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IEC/DPAS 63086-3-1:2022(E)
© ISO 2022
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ii
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CONTENTS
1. INTRODUCTION AND SCOPE .1
2. NORMATIVE REFERENCES .2
3. TERMS AND DEFINITIONS .2
3.1 Air Cleaner .2
3.2 Background Concentratoion .3
3.3 Bacteria………………………… .3
3.4 Bacteriophage or Phage……………… .3
3.5 Bioaerosol .3
3.6 BSL .3
3.7 CADR (Clean Air Delivery Rate) .3
3.8 Colony Forming Unit (CFU) for bacteria and fungi .4
3.9 Device Under Test (DUT) .4
3.10 Fungi .4
3.11 Impaction .4
3.12 Impinger Method .4
3.13 Initial Concentration .4
3.14 Maximum Performance Mode .4
3.15 Microbes .4
3.16 Microbial Reduction .4
3.17 Natural Decay .5
3.18 Plaque Forming Units (PFU) .5
3.19 Virus .5
4. PRINCIPLE .5
5. APPARATUS AND MATERIALS. .5
5.1 Apparatus .5
5.1.1. Test Chamber .6
5.1.2. Nebulizer……………………………………………………………………….7
5.1.3. SKC Bio sampler for sampling of microbes .7
5.1.4. Flow Calibration……………… .8
5.1.5. Stand. .8
5.1.6. Autoclave .8
5.1.7. Incubator .8
5.1.8. Deep freezer .8
5.1.9. Microbiological safety cabinet class II. .8
5.1.10. Balance. .8
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5.1.11. Inoculating loop .8
5.1.12. Petri dishes. .8
5.1.13. Disinfectant .8
5.1.14. UV-C Lamp……………… .8
5.1.15. pH-meter .8
5.1.16. Timer. .8
5.1.17. Stirrer .8
5.1.18. Recirculation fan .9
5.1.19. Refrigerator .9
5.2. Key Bioaerosols .9
5.2.1. Test Bacteria (Gram Positive) .9
5.2.2. Test Bacteria (Gram Negative) .9
5.2.3. Test Bacteria (Spore Forming).9
5.2.4. Test Bacteriophages (Non-enveloped) .10
5.2.5. Test Mold Spores .10
6. PREPARATION OF THE STOCK CULTURES AND WORKING CULTURES OF THE TEST
MICROBES .11
7. PROCEDURES.11
7.1. General .11
7.1.1 Electrical Supply .11
7.2. Step 1--Measurement of the concentration of airborne microbes, ci, without operating the air
cleaner .12
7.2.1. General .12
7.2.2. Preparation of the air cleaner and the test chamber .12
7.2.3. Measurement of microbial background concentration in the test chamber .13
7.2.4. Nebulizing test microbial suspension .13
7.2.5. Measurement of the initial concentration of airborne microbe inside the test chamber after
nebulizing .13
7.2.6. Measurement of the concentration of airborne microbe inside the test chamber after a
defined time .14
7.2.7. Post-test Actions .14
7.3. Step 2--Measurement of the concentration of airborne test microbe, ct, after operating the air
cleaner .15
8. CALCULATION AND EXPRESSION OF RESULTS .17
8.1. Conditions for a valid test .17
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8.1.1 Criteria for Elimination of Data Points from an AC-5 Run
8.2. Calculation of the concentration of airborne microbes .17
8.3. Reduction of Microbes .18
8.4. Calculating the Decay Constant .18
8.4.1. The Formula for Decay (Equation) .18
8.4.2. The Decay Constant using linear regression (Equation).19
8.5. Computation of the Standard Deviation Estimate for the Slope of One Regression Line
......................................................................................................................................19
8.6. Performance Calculation…………… .20
8.7 Calculation of the Standard Deviation Estimate of the CADR for a Single Test .20
9. TEST REPORT .21
10. QUALITY ASSURANCE .22
ANNEX A--TEST CHAMBER .23
ANNEX B--CALCULATION FOR TOTAL DECAY .25
ANNEX C--ADDITIONAL MICROBES .26
ANNEX D--Slit Sampler Questions .27
ANNEX E - CHAMBER SIZE EVALUATION ….28
ANNEX F - SUGGESTED TEST REPORT USING THIS METHOD.29
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International Electrotechnical Commission
Method for Assessing the Reduction Rate of Key Bioaerosols
by Portable Air Cleaners Using an Aerobiology Test Chamber
Aerobiology Test Chamber
FOREWORD
1) The International Electrotechnical Commission (IEC) is a worldwide organization for standardization
comprising all national electrotechnical committees (IEC National Committees). The object of the IEC is to
promote international co-operation on all questions concerning standardization in the electrical and
electronic fields. To this end and in addition to other activities, IEC publishes International Standards,
Technical Specifications, Technical Reports, Publicly Available Specifications (PAS) and Guides
(hereafter referred to as ‘IEC Publication(s)’. Their preparation is entrusted to technical committees; any
IEC National Committee interested in the subject dealt with may participate in this preparatory work.
International governmental and non-governmental organizations liaising with the IEC also participate in
this preparation. IEC collaborates closely with the International Organization for Standardization (ISO) in
accordance with conditions determined by agreement between the two organizations.
2) The formal decisions or agreements of IEC on technical matters express, as nearly as possible, an
international consensus of opinion on the relevant subjects since each technical committee has
representation from all interested IEC National Committees.
3) IEC Publications have the form of recommendations for international use and area accepted by IEC
National Committees in that sense. While all reasonable efforts are made to ensure that the technical
content of IEC Publications is accurate, IEC cannot be held responsible for the way in which they are
used or for any misinterpretation by any end user.
4) In order to promote international uniformity, IEC National Committees undertake to apply IEC Publications
transparently to the maximum extent possible in their national and regional publications. Any divergence
between any IEC Publication and the corresponding national or regional publication shall be clearly
indicated in the latter.
5) IEC provides no marking procedure to indicate its approval and cannot be rendered responsible for any
equipment declared to be in conformity with an IEC Publication.
6) All users should ensure that they have the latest edition of this publication.
7) No liability shall attach to IEC or its directors, employees, servants, or agents including individual experts
and members of its technical committees and IEC National Committees for any personal injury, property
damage or other damage of any nature whatsoever, whether direct or indirect, or for costs (including legal
fees) and expenses arising out of the publication, use of, or reliance upon, this IEC Publication or any
other IEC Publications.
8) Attention is drawn to the Normative references cited in this publication. Use of the referenced publications
is indispensable for the correct application of this publication.
9) Attention is drawn to the possibility that some of the elements in this IEC Publication may be the subject
of patent rights, IEC shall not be held responsible for identifying any or all such patent rights.
A PAS is a technical specification not fulfilling the requirements for a standard but made available to the
public.
IEC PAS 63086-3-1 has been prepared by the Association of Home Appliance Manufacturers (AHAM) and
processed by IEC technical committee 59, Subcommittee 59N: Performance of household and similar
electrical appliances. It is based on ANSI/AHAM AC-5-2022.
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The text of this PAS is based on the following document
59N/XXX/DPAS
This PAS was approved for publication by the P-members of the committee concerned as indicated in the
following document
Report on voting
59N/XXX/RVDPAS
Following publication of this PAS, which is a pre-standard publication, the technical committee or
subcommittee concerned will transform it into an International Standard.
This PAS shall remain valid for an initial maximum period of 3 years starting from the publication date. The
validity may be extended for a single 3-year period, following which it shall be revised to become another type
of normative document, or shall be withdrawn.
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2 INTRODUCTION
4 This Publicly Available Specification (PAS) contains test procedures for measuring the reduction by
5 the air cleaner of micro-organisms suspended in the air in the specified test chamber. It also
6 prescribes a method for measuring the operating power and stand-by power of the air cleaner. The
7 test procedures may be applied to any brand or model of household and similar electrical air cleaners
8 within the stated confines of the standard’s limits of measurability for measuring performance.
10 The annexes to this PAS are included for informative purposes only unless the annexes are noted as
11 normative.
13 Warning—The tests given in this document shall be performed by expert staff trained to handle
14 microorganism-related techniques and in properly equipped laboratories under the supervision of a
15 skilled microbiologist. Some of the test micro-organisms might be facultative pathogens for humans,
16 animals and plants and requires a laboratory of an appropriate bio-safety level. National and
17 international safety procedures for working with infectious biomaterials shall be followed to prevent
18 any contamination of laboratory staff, apparatus, working place or environment in compliance with
19 national standards or regulations. This document does not purport to address all of the safety
20 aspects, if any, associated with its use. It is the responsibility of the user to establish appropriate
21 safety and health practices and ensure compliance with any national, regional or international
22 regulatory conditions.
24 This PAS may involve hazardous materials, operations and equipment. This PAS does not purport to
25 address all of the safety problems associated with its use. It is the responsibility of whoever uses this
26 PAS to consult and establish appropriate safety and health practices and determine the applicability
27 of any regulatory limitations prior to use.
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29 International Electrotechnical Commission
31 Method for Assessing the Reduction Rate of Key Bioaerosols by
32 Portable Air Cleaners Using an Aerobiology Test Chamber Air
33 Cleaners Using an Aerobiology Test Chamber
35 INTRODUCTION
36 Indoor air free of harmful microbes is important to the health of occupants. This is
37 particularly relevant with regard to increased time spent indoors.
38 Air cleaners are used to reduce the concentrations of microorganisms in indoor air.
39 The efficiency of such air cleaners to reduce airborne microorganisms can be assessed in test
40 chambers at controlled air temperature and relative air humidity.
41 1. SCOPE
42 This document specifies a method to evaluate the capability of portable household air
43 cleaners to reduce the concentration and viability of key experimentally generated
44 bioaerosols in a specified chamber.
45 The test is applicable to portable air cleaners commonly used in single room spaces such
46 as those based on mechanical filtration, ultraviolet (UV), ionizers, photocatalytic
47 oxidation, and ozone generators in-unit technology.
49 If the air cleaner does not claim to have the function of reducing microorganisms, this
50 standard may not be applicable unless it is being used to simply evaluate the
51 performance.
53 This document deals with measurement procedures regarding the reduction of the
54 microbial contamination related to electrical air cleaner appliances for household and
55 similar use.
57 This document does not apply to appliances intended to be used in medical, veterinary, or
58 pharmaceutical applications.
60 This document does not address sanitization, disinfection, or sterilization measures.
62 This document does not support, by itself any health-related claims or conclusions about
63 prevention or treatment of a disease or health improvement.
65 Note: IEC 63086-3-1 is created for Household and Similar Electrical Air Cleaners and is
66 not intended to conflict with or replace standards for commercial or industrial consumers.
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69 Note: In this standard, we do not suggest performance test methods that measure the by-
70 products of either the interaction between microbes or between the air cleaner and the
71 microbes tested in this standard. The formation of by-products is an important subject.
72 The subject of measuring by-products is under study, and AHAM will address this in
73 future documents.
75 Note: This standard does not apply to appliances intended for use in medical treatment
76 locations, such as surgical suites, laboratories, medical treatment rooms, etc.
77 2. NORMATIVE REFERENCES
78 The following documents are referred to in the text so that some or all of their content
79 constitutes the requirements of this document. Only the edition cited applies for dated
80 references, and the latest edition of the referenced document (including any amendments)
81 applies for undated references.
82 ASTM E741-11(2017): Standard Test Method for Determining Air Change in a Single Zone by
83 Means of a Tracer Gas Dilution
84 ASTM E3273-21: Standard Practice to Assess Microbial Decontamination of Indoor Air using
85 an Aerobiology Chamber
86 GB21551.3-2010: Antibacterial and cleaning function for household and similar electrical
87 appliances – Particular requirements for air cleaner
88 ISO 3696: Water for analytical laboratory use — Specification and test methods
89 ISO 16000-9: Indoor air — Part 9: Determination of the emission of volatile organic compounds
90 from building products and furnishing — Emission test chamber method
91 ISO 16000-36: Standard method for assessing the reduction rate of culturable airborne bacteria
92 by air cleaners using a test chamber
93 JEMA 1467-2015: Appendix D - Evaluation test for removing performance of floating virus
94 JEMA 1467-2015: Appendix E - Evaluation test for restraining performance of indoor adhered
95 virus
96 JEMA 1467-2015: Appendix F - Evaluation test for restraining performance of virus caught by
97 the filter
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98 3. TERMS AND DEFINITIONS
99 3.1 Air Cleaner
100 electrically powered household or similar, appliance that employs one or multiple
101 technologies to reduce, destroy, and/or inactivate one or more types of indoor air
102 pollutants [Source: IEC 63086-1 modified]
103 Note to entry: the term Air Purifier is defined as an Electrically powered
104 device that is basically built of a fan and a set of components possessing
105 the ability to capture and/or (partially or totally) destroy air pollutants
106 [Source: ISO 16000-36] but PAS 63086-3-1 has chosen to not use this
107 term in this document as it may not be possible to totally destroy an air
108 pollutant.]
109 3.2 Background Concentration
110 quantity of microbes in the chamber after the chamber has undergone cleaning
111 and prior to any testing or addition of microbes via nebulization
112 3.3 Bacteria
113 prokaryotic, single-celled, microscopic organism with peptidoglycan cell wall
114 3.4 Bacteriophage or phage
115 group of viruses that infect bacteria or fungi
116 3.5 Bioaerosol
117 airborne particle that is composed of or derived from biological matter (such as a
118 bacterial cell, fungal or bacteria spore, virus, or endotoxin)
119 3.6 Biological Safety Levels (BSL)
120 series of protections relegated to autoclave-related activities that take place in
121 particular biological labs
122 Note to entry: This includes individual safeguards designed to protect laboratory
123 personnel, as well as the surrounding environment and community. For BSL
124 level expectations, a lab should follow the most recent version of the WHO
125 Laboratory Biosafety Manual, the CDC Biosafety in Microbiological and
126 Biomedical Laboratories (BMBL) or the Canadian Biosafety Standards and
127 Guidelines.
128 3.7 CADR (Clean Air Delivery Rate)
129 measure of air cleaner performance by this test procedure
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131 Note to entry: Clean Air Delivery Rate (CADR) is defined as the measure of the
132 delivery of contaminant-free air, within the defined particle size range, by an air
133 cleaner, expressed in cubic feet per minute (cfm) or cubic meters per hour. Clean
134 Air Delivery Rates are the rates of contaminant reduction in the test chamber
135 when the air cleaner is turned on, minus the rate of natural decay when the air
136 cleaner is not running, multiplied by the volume of the test chamber as measured
137 in cubic feet or cubic meters (see Section 8.5). CADRs values are always the
138 measurement of an air cleaner performance as a complete system, and they have
139 no linear relationship to air movement per se or to the characteristics of any
140 particular particle removal methodology.
142 Note to entry: For this standard we use the designation of m-CADR which is the
143 clean air delivery rate for microbes.
144 3.8 Colony Forming Units (CFU) for bacteria and fungi
145 unit of measurement by which the number of culturable microbes (Bacteria and
146 fungi) is expressed
147 3.9 Device Under Test (DUT)
148 test sample of the air cleaner undergoing examination
149 3.10 Fungi
150 multicellular eukaryotic organisms without chlorophyll and with cell walls
151 3.11 Impaction
152 sampling of the airborne microbe by inertial separation on a semisolid agar surface
153 3.12 Impinger method
154 glass or plastic device for the collection of air samples into a liquid medium
155 through a scrubbing action.
157 Note to entry: The liquid volume is subsequently utilized for dilution and
158 inoculation of counting plates.
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159 3.13 Initial Concentration
160 concentration of microbes inside the chamber immediately at the start time of
161 sampling of either the natural decay or the total decay
162 3.14 Maximum Performance Mode
163 through manual operation the DUT is set to the highest flow rate with all air
164 cleaning functions switched on, set to maximum, where applicable, and with all
165 filters in place
166 Note to entry: If the DUT has zero flow rate, the m-CADR is measured with all
167 air cleaning functions switched on.
168 3.15 Microbes (also known as Microorganisms)
169 microscopic living beings that cannot be seen with the naked eye, including
170 bacteria, protozoa, viruses and some fungi/fungal components
172 Note to entry: They are common in the environment as well as in/on our own
173 bodies.
174 3.16 Microbial Reduction
175 reduction rate of viable microbe is measured by comparing the concentration of
176 the microbe after nebulizing a microbial suspension inside the chamber with the
177 concentration determined after a defined running time (testing time) of the air
178 cleaner
179 Note to entry: The microbial reduction rate is expressed as natural log reduction
180 over time.
181 3.17 Natural Decay
182 rate of reduction of the airborne concentration of viable microbiological
183 contaminants as measured without an air-cleaning device operating in an
184 aerobiology chamber
186 Note to entry: The natural decay rate is expressed as natural log reduction
187 over time.
189 3.18 Plaque Forming Units (PFU)
190 Unit of measurement by which the number of viable viruses is expressed
191 3.19 Virus
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192 group of microorganisms with a simple structure composed of RNA or DNA and
193 protein outer coat which are specialized in intracellular infection and replication
194 4. PRINCIPLE
195 The efficiency of air cleaners is tested using one or more nebulized and homogeneously
196 distributed microbial suspensions inside an enclosed test chamber at controlled air
197 temperature and relative air humidity. The efficacy is calculated by the reduction rate of
198 the test microbe in a defined period of time, considering the rate of natural decay of the
199 test microbe.
200 5. APPARATUS AND MATERIALS
201 5.1 Apparatus
202 Informative Note - As a wide variety of specialized pieces of equipment exist and
203 are commercially available, the following list gives only the preferred list of
204 equipment that have the desired primary critical characteristics. Alternates are
205 allowed when they have been shown to be equivalent. Equivalency specifications
206 or data should be included showing the alternate equipment can be considered
207 equivalent.
208 5.1.1 Test chamber
209 The chamber shall be constructed to the following characteristics:
210 • Be accepted by OSHA (U.S. Occupational Safety and Health
211 Administration) or other national bodies;
212 • The chamber size is 30 ± 1,5 m ; Height = (2,5 ± 0,1) m. The width shall
213 be within 85 % and 100 % of the length. See Appendix F for use of a
214 chamber of alternate size;
215 • The walls should be made from a suitable smooth non-porous material that
216 emits minimal levels of volatile organics, is corrosion-resistant, and is
217 repeatedly washable (i.e., constructed of stainless steel, epoxy, glass or
218 other documented nonreactive material with minimum volatile organic
219 hydrocarbon emission potential). The material should not quench
220 ionization, be non-reflective for visible and ultraviolet light (which is
221 measured as between 5 and 20 % reflectance at the operational wavelength
222 of the device under test), and be well-grounded;
223 • It shall maintain sufficient airtight capacity. The test chamber air
224 exchange rate is to be less than 0,05 air changes per hour (ACH) as
225 determined by ASTM E 741 (Standard Test Method for Determining Air
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226 Change in a Single Zone by Means of a Tracer Gas Dilution) or an
227 equivalent method;
228 • The test environment shall be kept clean and free from extraneous
229 microbial contamination. It shall have a suitable environmental control
230 system to maintain a controlled level of air temperature and humidity. To
231 achieve this, the test chamber should include the following:
232 • A system capable of removing contamination and maintaining
233 aseptic condition inside the chamber, such as an UV lamp;
234 • A facility to transfer items into and out of the chamber without
235 cross-contamination (this can include a special system, such as a
236 glove box, etc.);
237 • The chamber may be fitted with an anteroom to allow for staging;
238 • A facility to control the power inside the chamber from outside;
239 • The chamber shall be equipped such that tests can be witnessed
240 externally;
241 • A facility to generate an aerosol of test microbe inside the
242 chamber and to ensure its homogeneity (this can be achieved by
243 using a nebulizing inlet through which microbes are nebulized,
244 connected to an atomizing nozzle in the chamber, with a fan to
245 ensure homogeneous distribution of the microbe inside the
246 chamber);
247 • A sampling port should be 120 cm high from the floor. The port
248 should be a minimum of 30,5 cm from the wall and a minimum
249 of 91,4 cm away from the device and out of the airflow of the air
250 cleaner exhaust or intake. See Section 7.2.2. for unit
251 positioning;
252 • An air conditioning system inside the chamber capable of
253 controlling air temperature and relative humidity in a stable and
254 precise manner; the air conditioning system shall be switched off
255 during the test. No other external temperature or humidity
256 manipulating equipment for the chamber shall be operated
257 during the test
258 the initial test air temperature and acceptable range
259 of variation shall be (20 ± 3) °C;
260 the initial test relative humidity and acceptable
261 range of variation shall be (50 ± 10) %;
262 The test chamber shall be equipped to continuously
263 monitor and record humidity and temperature
264 • A facility to use negative pressure airflow to flush the chamber
265 post-testing;
267 • A filter to prevent contamination from the outside during
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268 ventilation. A HEPA filter is recommended to be used in the
269 incoming and outgoing air to prevent lab contamination from
270 entering the chamber or residual microbes contaminating the
271 surrounding space.
273 See graphics of an example test chamber in Annex A.
274 5.1.2 Nebulizer
275 The nebulizer shall be capable of nebulizing microbial suspensions into
276 particles (0,05 µm to 5 µm) to produce, as far as possible, discrete
277 particles. It typically comprises a pump to generate a defined air pressure
278 to nebulize, a clean air supplying unit and a dehumidifier to remove excess
279 water from the generated culture medium. A compressed air cylinder can
280 also be used to operate the nebulizer.
281 • Collison 6-jet nebulizer (BGI Inc. Waltham MA) driven by
282 purified filtered house air supply or equivalent
284 5.1.2.1 Nebulizer Fluid
285 The nebulizer reservoir should be filled with a combination of test
286 microbial suspension, deionized water, antifoaming agent and
287 phosphate-buffered saline (PBS). The concentrations in the
288 mixture the lab uses should be specified in the report (see Section
289 9).
290 5.1.3 SKC Bio sampler or equivalent for the sampling of microbes
291 The recommended impinger tube size is 20 mL.
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293 Fig 2 – SKC Biosampler
294 5.1.4 Flow calibration – SKC Checkmate or equivalent
295 5.1.5 Stand, to position the sample tube per 5.1.1.
296 5.1.6 Autoclave, thermostatically controlled at (121 ± 3) °C and pressure of
297 (103 ± 5) kPa.
298 5.1.7 Incubator, thermostatically controlled at the appropriate temperature
299 depending on the microbe under test.
300 5.1.8 Deep freezer, thermostatically controlled at (–80 ± 10) °C.
301 5.1.9 Microbiological safety cabinet class II.
302 5.1.10 Balance, capable of weighing to ±0,01 g.
303 5.1.11 Inoculating loop, 4 mm in ring diameter, sterile.
305 5.1.12 Petri dishes, vented, sterile, 90 mm to 150 mm diameter.
307 5.1.13 Disinfectant, ethanol (70 % volume fraction) or bleach (3 % by volume).
309 5.1.14 UV-C Light (254 to 265 nm wavelength with a minimum of 2mW/cm )
311 5.1.15 pH-meter, capable of measuring to ±0,2 unit.
313 5.1.16 Timer.
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315 5.1.17 Stirrer – A high volume ceiling fan used to mix the test chamber during
316 contaminant aerosol generation.
318 Example - Ceiling Fan Available from W.W. Grainger:
319 3 blade, 0,91 m, ceiling fan 395 RPM
320 Stock No. 36201 or equivalent
321 Amps 0,65, Volts 110-120
322 Weight 9,07 kg (20 lbs.)
323 Informative note – it is acceptable to use multiple fans replacing the ceiling
324 fan. Evaluation is needed to show the mixing is equivalent. Make sure the multiple
325 fans are not artificially impacting the decay rate.
327 5.1.18 Recirculation fan - A fan capable of producing between 50 and 250 cfm
328 and used for the purpose of maintaining a homogeneous environment
329 within the test chamber
331 5.1.19 Refrigerator (5 ± 3 °C) for storage of culture media, culture plates and
332 reagents.
335 5.2 Key Bioaerosols
336 The preferred microbe for testing is noted below, and their associated American
337 Type Culture Collection (ATCC) catalog numbers are noted below
338 (www.atcc.org). See Annex C for Alternate microbes and Annex D for
339 justification for the particles listed below. Any of the alternates in Annex C can
340 also be used but need to be noted in the report.
341 • For testing of bacteria, interactions are different so we recommend
342 testing more than one. For example: U.S. EPA and this test
343 method recommends both a Gram-Positive and a Gram-Negative
344 (see 5.2.1 & 5.2.2). 5.2.3 is used for bacterial endospores.
345 • For testing of virus, susceptibility based on their relative resistance
346 to inactivation is different so we recommend testing more than one
347 type of bacteriophage as surrogates for human pathogenic viruses
348 (see U.S. EPA classification as noted in Annex D).
349 o See 5.2.4.1 and the alternates in the Annex C.
350 • For testing of mold, we are recommending mold spore (see 5.2.5)
352 5.2.1 Test Bacteria (Gram-Positive)
353 5.2.1.1 Staphylococcus epidermidis (BSL-1) ATCC 12228
354 5.2.2 Test Bacteria (Gram-Negative)
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355 5.2.2.1 Acinetobacter baumannii (BSL-2) ATCC 19606
356 5.2.3 Test Bacteria (Spore Forming)
357 5.2.3.1 Geobacillus stearothermophilus (BSL-1) ATCC 12980
358 5.2.4 Test Bacteriophages (Non-enveloped)
359 5.2.4.1 MS2 (preferred) (BSL-1) ATCC 15597-B1
360 with host Escherichia coli ATCC 15597
361 5.2.5 Test Mold Spores
362 5.2.5.1 Aspergillus brasiliensis (BSL-1) ATCC 16404
364 For specific questions, other microbes may be used. All strains used shall be listed in
365 the test report.
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367 6. Preparation of the stock cultures and working cultures of the test microbes
368 Culture Method is per ATCC recommendations or equivalent for a defined culture
369 method for each microbe. The same agar from the same source should be used
370 throughout the same evaluation.
371 7. Procedures
372 WARNING — The test method given in this document shall be performed by expert staff
373 trained and certified to handle microorganism-related techniques and in properly equipped
374 laboratories under the control of a skilled microbiologist. Some of the suggested test
375 micro-organisms can be pathogens for humans, other animals and plants. Great care should
376 be taken in the disposal of all incubated test materials and surfaces exposed to
377 aerosolization of the micro-organisms. National and international safety procedures for
378 working with infectious biomaterials shall be followed to prevent any contamination of
379 laboratory staff, apparatus, working place or environment. The examination and
380 preparation of the cultures should be carried out in a microbiological safety cabinet or
381 chamber at the appropriate bio-safety level (BSL) in compliance with national standards
382 or regulations. This document does not purport to address all of the safety aspects, if any,
383 associated with its use. It is the responsibility of the user to establish appropriate safety
384 and health practices and ensure compliance with any national, regional or international
385 regulatory conditions.
386 7.1 General
387 Prevent any extraneous microbial contamination by preparing and
388 handling the test microbes using a certified biological safety cabinet
389 (BSC). The lab should also have a laboratory safety plan that is adequate
390 to handle the specified BSL for the chosen bioaerosols.
391 The test is performed in two steps. In Step 1 (see Section 7.2) the concentration of
392 the nebulized test microbe in the chamber's air is measured without operating the
393 air cleaner, then in Step 2 (see Section 7.3) the concentration of the nebulized
394 test microbe in the chamber's air is measured with the air cleaner operating.
395 The test is only valid if the conditions in 8.2 are met. If these conditions
396 are not met, the test (Step 1 and Step 2) shall be repeated.
397 7.1.1 Electrical Supply
398 Standard frequency and voltage for the m-CADR testing and operating
399 power test are listed in Sections 7.1.1.1 and 7.1.1.2. Other frequencies
400 and voltages may be used to produce m-CADR values. The specific
401 electrical supply conditions shall be concurrently reported with the
402 applicable m-CADR values.
404 7.1.1.1 Frequency
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405 Operate air cleaner at Nameplate frequency ± 1% Hertz
406 7.1.1.2 Voltage
407 Operate air cleaner at Nameplate voltage ± 1% Volts.
408 7.2 Step 1 — Measurement of the concentration of airborne microbes, c , without
i
409 operating the air cleaner
410 7.2.1 General
411 In Step 1, the concentration of the airborne test microbe is measured
412 without operating the air cleaner.
413 7.2.2 Preparation of the air cleaner and the test chamber
414 Prior to the initial test, the air cleaner motor must be properly broken in
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