ISO 18071:2016

Fine ceramics (advanced ceramics, advanced technical ceramics) — Determination of antiviral activity of semiconducting photocatalytic materials under indoor lighting environment — Test method using bacteriophage Q-beta

ISO 18071:2016 specifies the determination of the antiviral activity of materials that contain indoor-light-active photocatalytic materials or have indoor-light-active photocatalytic films on the surface by a test method that measures the infectivity titre of bacteriophage Q-beta after illumination with indoor light. NOTE In the test method, the surrogate microbe is bacteriophage Q-beta, intended as a model for influenza viruses. It is intended for use with different kinds of indoor-light-active photocatalytic materials used in construction materials, in flat sheet, board or plate shape that are the basic forms of materials for various applications. It does not include powder, granular or porous indoor-light-active photocatalytic materials. It is applicable to indoor-light-active photocatalytic materials produced for an antiviral applications. Other types of performance of indoor-light-active photocatalytic materials, i.e. antibacterial activity, antifungal activity, decomposition of water contaminants, self-cleaning, antifogging and air purification, are not determined by this method.

Céramiques techniques — Détermination de l'activité antivirale des matériaux photocatalytiques semi-conducteurs dans un environnement d'éclairage intérieur — Méthode d'essai utilisant un bactériophage Q-béta

General Information

• Status: Published
• Publication Date: 14-Jul-2016
• ICS: 81.060.30 - Advanced ceramics
• Technical Committee: ISO/TC 206 - Fine ceramics
• Drafting Committee: ISO/TC 206/WG 9 - Photocatalysis
• Current Stage: 9093 - International Standard confirmed
• Start Date: 03-Dec-2021
• Completion Date: 13-Dec-2025

Relations

• Consolidated By: ISO 4884:2019 - Hardmetals — Sampling and testing of powders using sintered test pieces
• Effective Date: 06-Jun-2022

Standards Content (Sample)

INTERNATIONAL ISO
STANDARD 18071
First edition
2016-07-15
Fine ceramics (advanced ceramics,
advanced technical ceramics) —
Determination of antiviral activity
of semiconducting photocatalytic
materials under indoor lighting
environment — Test method using
bacteriophage Q-beta
Céramiques techniques — Détermination de l’activité antivirale
des matériaux photocatalytiques semi-conducteurs dans un
environnement d’éclairage intérieur — Méthode d’essai utilisant un
bactériophage Q-béta
Reference number
©
ISO 2016
© ISO 2016, Published in Switzerland
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
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ii © ISO 2016 – All rights reserved

Contents Page
Foreword .v
Introduction .vi
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Symbols . 2
5 Principle . 3
6 Materials . 3
6.1 Strains and preparation for tests . 3
6.1.1 Strains . 3
6.1.2 Bacteria preparation. 4
6.1.3 Bacteriophage preparation . 4
6.2 Media . 5
6.2.1 General. 5
6.2.2 1/500 Nutrient broth (1/500 NB) . 5
6.2.3 Calcium solution . 5
6.2.4 LB broth with calcium . 5
6.2.5 Agar powder . 5
6.2.6 LB agar . 5
6.2.7 Bottom agar plate (LB agar plate with calcium) . 6
6.2.8 Top agar . 6
6.2.9 Soybean-casein digest broth with lecithin and polysorbate 80 (SCDLP). 6
6.2.10 Peptone saline solution . 6
7 Apparatus and equipment . 6
7.1 Test equipment . 6
7.2 Cover film . 7
7.3 Moisture preservation glass . 7
7.4 Glass tube or glass rod . 7
7.5 Paper filter . 7
7.6 Light source . 7
7.7 UV sharp cut-off filter . 8
7.7.1 Condition A (under 400 nm cut-off condition) . 8
7.7.2 Condition B (under 380 nm cut-off condition) . 8
7.8 Illuminance meter . 8
7.9 Centrifuge . 8
7.10 Sterilized syringe filter unit . 8
8 Test piece . 8
9 Procedure. 8
9.1 General . 8
9.2 Procedure for preparation of bacteria suspension . 9
9.3 Procedure of preparation of test bacteriophage solution . 9
9.4 Procedure of test for indoor-light-active photocatalytic antiviral activity .10
9.5 Indoor lighting condition .10
9.6 Measurement of titre of bacteriophage .11
10 Calculation .12
10.1 General .12
10.2 Test requirement fulfilment validation .12
10.3 Indoor-light-active photocatalyst antiviral activity value calculation .13
10.4 Antiviral activity value calculation without indoor-light-active photocatalyst .13
11 Test report .14
Bibliography .15
iv © ISO 2016 – All rights reserved

Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out
through ISO technical committees. Each member body interested in a subject for which a technical
committee has been established has the right to be represented on that committee. International
organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.
ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of
electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www.iso.org/patents).
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment,
as well as information about ISO’s adherence to the World Trade Organization (WTO) principles in the
Technical Barriers to Trade (TBT) see the following URL: www.iso.org/iso/foreword.html.
The committee responsible for this document is ISO/TC 206, Fine ceramics.
Introduction
This International Standard applies to testing the antiviral activity of indoor-light-active photocatalytic
ceramics and other materials produced by either coating or mixing of a light-active photocatalyst.
The International Standard for testing the antibacterial activity of photocatalytic materials has been
published as ISO 27447 and the International Standard for testing the antibacterial activity of indoor-
light-active photocatalytic materials has been published as ISO 17094. The International Standard for
determination of antiviral activity of semiconducting photocatalytic materials has also been published
as ISO 18061.
The test method for cloths or textiles is not included in this International Standard because of lack of
indoor-light-active photocatalytic cloths or textiles. When the indoor-light-active photocatalytic cloths
or textiles with antiviral activity using indoor-light-active photocatalytic activity have been developed,
a test method for indoor-light-active photocatalytic cloths or textiles will be proposed with the glass
adhesion method in ISO 27447.
vi © ISO 2016 – All rights reserved

INTERNATIONAL STANDARD ISO 18071:2016(E)
Fine ceramics (advanced ceramics, advanced technical
ceramics) — Determination of antiviral activity of
semiconducting photocatalytic materials under
indoor lighting environment — Test method using
bacteriophage Q-beta
WARNING — Only personnel trained in microbiological techniques should carry out tests.
1 Scope
This International Standard specifies the determination of the antiviral activity of materials that
contain indoor-light-active photocatalytic materials or have indoor-light-active photocatalytic films
on the surface by a test method that measures the infectivity titre of bacteriophage Q-beta after
illumination with indoor light.
NOTE In the test method, the surrogate microbe is bacteriophage Q-beta, intended as a model for influenza
viruses.
This International Standard is intended for use with different kinds of indoor-light-active photocatalytic
materials used in construction materials, in flat sheet, board or plate shape that are the basic forms of
materials for various applications. It does not include powder, granular or porous indoor-light-active
photocatalytic materials.
This International Standard is applicable to indoor-light-active photocatalytic materials produced for
an antiviral applications. Other types of performance of indoor-light-active photocatalytic materials,
i.e. antibacterial activity, antifungal activity, decomposition of water contaminants, self-cleaning,
antifogging and air purification, are not determined by this method.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any amendments) applies.
ISO 14605, Fine ceramics (advanced ceramics, advanced technical ceramics) — Light source for testing
semiconducting photocatalytic materials used under indoor lighting environment
ISO 27447, Fine ceramics (advanced ceramics, advanced technical ceramics) — Test method for antibacterial
activity of semiconducting photocatalytic materials
ISO 80000-1, Quantities and units — Part 1: General
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
photocatalyst
substance that carries out many functions based on oxidization and reduction reactions under
photoirradiation, including decomposition and removal of air and water contaminants, deodorization,
and antiviral, antibacterial, antifungal, self-cleaning and antifogging actions
3.2
indoor-light-active photocatalyst
substance that reacts with artificial light source for general lighting service (i.e. indoor lighting
environment)
3.3
indoor lighting environment
environment with artificial light source for general lighting service
Note 1 to entry: Does not include sunlight.
3.4
indoor-light-active photocatalytic materials
materials in which or on which the indoor-light-active photocatalyst is added by coating, impregnation,
mixing, etc.
3.5
antiviral
condition decreasing the infectivity of viruses on the surface of materials
3.6
bacteriophage
type of virus which infects bacteria
Note 1 to entry: The bacteriophage used in this International Standard is Q-beta that is one of F-specific RNA
phages. The bacteriophage Q-beta is not pathogenic to humans and animals, but serves to simulate Influenza
viruses that are pathogenic to humans.
3.7
plaque
visible, clear area which is theoretically the result of infection and lysis of host cells by a single viable
bacteriophage
3.8
indoor-light-active photocatalyst antiviral activity value
difference between the logarithms of the total number of bacteriophage plaques on photocatalytic
treated materials after indoor light illumination and on non-treated materials after indoor light
illumination
Note 1 to entry: This value includes the decrease of number of bacteriophage plaques without indoor light
illumination.
3.9
indoor-light-active photocatalyst antiviral activity value for indoor light illumination
difference between the logarithms of the total number of bacteriophage plaques on photocatalytic treated
materials after indoor light illumination and on photocatalytic treated materials kept in a dark place
4 Symbols
A average of titre of bacteriophage on non-treated specimens, just after inoculation
B average of titre of bacteriophage on non-treated specimens, after being kept in a dark place
D
B average of titre of bacteriophage on non-treated specimens, after indoor light illumination
F-L
of intensity L under condition F
C average of titre of bacteriophage on indoor-light-active photocatalytic treated specimens,
D
after being kept in a dark place
2 © ISO 2016 – All rights reserved

C average of titre of bacteriophage on indoor-light-active photocatalytic treated specimens,
F-L
after indoor light illumination of intensity L under condition F
D dilution factor
F
F type of UV cut-off condition (condition A or condition B)
L illuminance of indoor light
Logmax maximum logarithmic value of titre of bacteriophage
Logmean average logarithmic value of titre of bacteriophage for three non-treated specimens
Logmin minimum logarithmic value of titre of bacteriophage
N titre of bacteriophage (plaque forming unit)
V antiviral activity value without indoor-light-active photocatalyst, after being kept in a dark
D
place on a testing material
V indoor-light-active photocatalyst antiviral activity value, after indoor light illumination at
F-L
a constant intensity (F-L) on an indoor-light-active photocatalytic material
ΔV indoor-light-active photocatalyst antiviral activity value with indoor light illumination
Z average number of plaques in two Petri dishes
5 Principle
The test method is suitable for use in development, comparison, quality assurance, characterization,
reliability and design data generation of indoor-light-active photocatalytic materials. The method is
used to obtain the antiviral activity of indoor-light-active photocatalytic materials by the contact of a
specimen with bacteriophage under indoor lighting condition. The method is suitable for use with flat
sheet, board or plate-shaped materials.
The specimen of indoor-light-active photocatalytic treated material is inoculated with bacteriophage
suspension and exposed to light for a specified period. Following exposure, the test suspension is
removed and measured by the plaque forming method with Escherichia coli which is sensitive to
bacteriophage Q-beta. The results obtained are compared with those obtained from inoculated
specimens of non-photocatalytic treated material exposed to light under identical conditions to the
treated material and to those obtained from inoculated specimens of both photocatalytic treated and
non-treated material kept in the dark for the same period of time.
NOTE This International Standard is adapted from the common methodological concept for ISO 18061.
Namely, the same apparatus without light source (see 7.6), UV sharp cut-off filter (see 7.7), and test piece size,
similar procedure and calculation are adapted between this International Standard and ISO 18061. Therefore,
ISO 18061 is recommended to be used as reference during actual test of this International Standard.
6 Materials
6.1 Strains and preparation for tests
6.1.1 Strains
The strains to be used in the test shall be the same as or equivalent to those described in Table 1 and
supplied by an entity that is registered under the World Federation for Culture Collections or the Japan
Society for Culture Collections. Aseptic manipulations using microorganisms can be performed in an
appropriate safety cabinet.
Table 1 — Bacteriophage and bacteria strains to be used in test
Species Strain number Organization for the collection
Bacteriophage Q-beta ATCC 23631-B1 American Type Culture Collection
German Collection of Microorganisms and
DSM 13768
Cell Cultures (DSMZ)
NBRC 20012 NITE Biological Resource Center
Escherichia coli ATCC 23631 American Type Culture Collection
German Collection of Microorganisms and
DSM 5210
Cell Cultures (DSMZ)
NBRC 106373 NITE Biological Resource Center
NOTE ATCC23631-B1 and NBRC20012 are not strictly same, but they are from the same origin.
6.1.2 Bacteria preparation
a) Inoculate E. coli strain into a slant culture medium (6 ml to 10 ml of LB agar; see 6.2.6), incubate for
16 h to 24 h at (37 ± 1) °C and then store in the refrigerator at 5 °C to 10 °C.
b) Repeat subcultures within 1 month by replicating this process.
c) The slant culture shall not be used for further storing after 1 month.
d) The maximum number of subcultures from the original strain transferred by culture collection is 10.
NOTE In the case of bacteria stored in a deep freezer, the maximum number of subcultures from original
strain transferred by culture collection is 10.
6.1.3 Bacteriophage preparation
a) Introduce 25 ml of LB broth with calcium (see 6.2.4) into a conical flask of 300 ml and inoculate
with E. coli strain.
−1
b) Incubate for (18 ± 2) h at (37 ± 1) °C while shaking at (110 ± 10) min .
c) Pre-warm 25 ml of LB broth with calcium (see 6.2.4) in a 300 ml conical flask to 35 °C to 37 °C and
inoculate with 0,025 ml of the culture prepared under b).
d) Incubate as above condition until a bacterial concentration will be reached at (2,0 ± 1,0) × 10 cfu/ml.
This procedure should be carried out several times to establish the relationship between turbidity
measurements and colony counts. If sufficient data have been obtained, further work can be based on
turbidity measurements only.
e) Inoculate the bacterial culture with Q-beta from a stock solution to a final concentration
of approximately 2 × 10 pfu (plaque forming unit)/ml [multiplicity of infection (m.o.i.) is
approximately 0,1].
f) Incubate the inoculated bacterial culture for 4 h as under b).
g) Store the culture overnight at (4 ± 2) °C.
h) Pour the culture into centrifuge tubes and centrifuge for 20 min at (4 ± 2) °C at 10 000g.
i) Pipette the supernatant carefully to a sterilized tube.
j) Filter bacteriophage containing supernatant suspension through a sterilized syringe filter unit to
purify the bacteriophage solution.
k) Determine the titre of the bacteriophage stock solution (see 9.6) and store at (4 ± 2) °C.
4 © ISO 2016 – All rights reserved
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