Textiles - Determination of antibacterial activity of textile products (ISO 20743:2013)

This International Standard specifies quantitative test methods to determine the antibacterial activity of antibacterial all textile products including nonwovens. This International Standard is applicable to all textile products, including cloth, wadding, thread and material for clothing, home furnishings and miscellaneous goods regardless of the type of antibacterial agent used (organic, inorganic, natural or man-made) or the method of application (built-in, after-treatment or grafting). Based on the intended application and on the environment in which the textile product is to be used, the user can select the most suitable of the following three methods on determination of antibacterial activity: a) absorption method (an evaluation method in which test bacterial suspension is inoculated directly onto samples); b) transfer method (an evaluation method in which test bacteria are placed on an agar plate and transferred onto samples); c) printing method (an evaluation method in which test bacteria are placed on a filter and printed onto samples). The colony plate count method and the ATP (ATP = Adenosine Tri-phosphate) luminescence method are also specified for measuring the enumeration of bacteria.

Textilien - Bestimmung der antibakteriellen Wirksamkeit von textilen Produkten (ISO 20743:2013)

Die vorliegende Internationale Norm legt quantitative Prüfverfahren zur Bestimmung der antibakteriellen Wirkung antibakteriell behandelter textiler Erzeugnisse, einschließlich Vliesstoffe, fest.
Diese Internationale Norm ist anwendbar auf alle textilen Erzeugnisse, einschließlich Stoffe, Wattierung, Garn und Materialien für Bekleidung, Heimtextilien sowie verschiedene Waren, ungeachtet des Typs des verwendeten antibakteriellen Mittels (organisch, anorganisch, natürlich oder künstlich) oder des Anwendungs-verfahrens (eingebaut, Nachbehandlung oder Pfropfen).
Auf der Grundlage der beabsichtigten Anwendung und der Umgebung, für die das textile Erzeugnis zu verwenden ist, kann der Anwender aus den folgenden drei Verfahren zur Bestimmung der antibakteriellen Wirkung das geeignetste auswählen.
a)   Absorptionsverfahren (ein Bewertungsverfahren, bei dem die bakterielle Prüfsuspension direkt auf Proben geimpft wird);
b)   Übertragungsverfahren (ein Bewertungsverfahren, bei dem Prüfbakterien auf eine Agarplatte aufgebracht und auf Proben übertragen werden);
c)   Stempelverfahren (ein Bewertungsverfahren, bei dem Prüfbakterien auf ein Filter aufgebracht und auf Proben gestempelt werden).
Das Kolonie-Plattenzählverfahren und das ATP-Lumineszenzverfahren (ATP = Adenosintriphosphat) werden auch zum Messen der Bakterienzahl festgelegt.

Textiles - Détermination de l'activité antibactérienne des produits textiles (ISO 20743:2013)

L'ISO 20743:2013 spécifie des méthodes d'essai quantitatives permettant de déterminer l'activité antibactérienne de tous les produits textiles antibactériens, y compris les nontissés.
L'ISO 20743:2013 s'applique à tous les produits textiles, y compris l'étoffe, le rembourrage, le fil et les matériaux utilisés pour les vêtements, la literie, l'ameublement et divers articles, quel que soit le type d'agent antibactérien utilisé (organique, inorganique, naturel ou synthétique) ou quelle que soit la méthode d'application (intégration, post-traitement ou greffage).
Tenant compte de l'application prévue et de l'environnement dans lequel le produit textile est destiné à être utilisé, et également des propriétés de surface du textile, l'utilisateur peut choisir la plus adaptée des trois méthodes d'ensemencement suivantes pour la détermination de l'activité antibactérienne:
a) méthode par absorption (méthode d'évaluation dans laquelle la suspension bactérienne d'essai est ensemencée directement sur des éprouvettes);
b) méthode par transfert (méthode d'évaluation dans laquelle les bactéries d'essai sont placées sur une boîte de milieu gélosé, puis transférées sur des éprouvettes);
c) méthode par impression (méthode d'évaluation dans laquelle les bactéries d'essai sont placées sur un filtre, puis imprimées sur des éprouvettes).
La technique de dénombrement et la méthode de mesure par luminescence de l'ATP (adénosine triphosphate) sont également spécifiées pour le dénombrement des bactéries.

Tekstilije - Ugotavljanje protibakterijske aktivnosti na tekstilnih izdelkih (ISO 20743:2013)

Ta mednarodni standard določa kvantitativne preskusne metode za ugotavljanje protibakterijske aktivnosti na vseh tekstilnih izdelkih, vključno z vlaknovinami. Ta mednarodni standard se uporablja za vse tekstilne izdelke, vključno z blagom, vato, nitmi in materialom za oblačila, izdelki za opremljanje domov in drugo, ne glede na vrsto uporabljene protibakterijske učinkovine (organskega, neorganskega, naravnega ali kemičnega izvora) ali način uporabe (vgrajeno, naknadna obdelava ali cepljenje). Na podlagi nameravane uporabe in okolja, v katerem se bo tekstilni izdelek uporabljal, lahko uporabnik izbere najbolj primerno izmed naslednjih treh metod za določanje protibakterijske aktivnosti: a) metodo absorpcije (metoda ocenjevanja, pri kateri je testna suspenzija bakterij vcepljena neposredno na vzorce); b) metoda prenosa (metoda ocenjevanja, pri kateri se testne bakterije razlijejo na ploščo gojišča in prenesejo na vzorce); c) metoda tiskanja (metoda ocenjevanja, pri kateri se testne bakterije razlijejo na filter in natisnejo na vzorce). Metoda za štetje kolonij in luminescenčna metoda z ATP-jem (ATP = adenozin trifosfat) sta tudi določeni za merjenje števila bakterij.

General Information

Status
Withdrawn
Publication Date
05-Nov-2013
Withdrawal Date
13-Jul-2021
Current Stage
9900 - Withdrawal (Adopted Project)
Start Date
14-Jul-2021
Due Date
06-Aug-2021
Completion Date
14-Jul-2021

RELATIONS

Buy Standard

Standard
SIST EN ISO 20743:2013
English language
40 pages
sale 10% off
Preview
sale 10% off
Preview

e-Library read for
1 day

Standards Content (sample)

SLOVENSKI STANDARD
SIST EN ISO 20743:2013
01-december-2013
1DGRPHãþD
SIST EN ISO 20743:2007

Tekstilije - Ugotavljanje protibakterijske aktivnosti na tekstilnih izdelkih (ISO

20743:2013)

Textiles - Determination of antibacterial activity of textile products (ISO 20743:2013)

Textilien - Bestimmung der antibakteriellen Wirksamkeit von textilen Produkten (ISO

20743:2013)

Textiles - Détermination de l'activité antibactérienne des produits textiles (ISO

20743:2013)
Ta slovenski standard je istoveten z: EN ISO 20743:2013
ICS:
07.100.99 Drugi standardi v zvezi z Other standards related to
mikrobiologijo microbiology
59.080.01 Tekstilije na splošno Textiles in general
SIST EN ISO 20743:2013 en

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
SIST EN ISO 20743:2013
---------------------- Page: 2 ----------------------
SIST EN ISO 20743:2013
EUROPEAN STANDARD
EN ISO 20743
NORME EUROPÉENNE
EUROPÄISCHE NORM
July 2013
ICS 07.100.99; 59.080.01 Supersedes EN ISO 20743:2007
English Version
Textiles - Determination of antibacterial activity of textile
products (ISO 20743:2013)

Textiles - Détermination de l'activité antibactérienne des Textilien - Bestimmung der antibakteriellen Wirkung

produits textiles (ISO 20743:2013) antibakteriell behandelter Erzeugnisse (ISO 20743:2013)

This European Standard was approved by CEN on 22 June 2013.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European

Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national

standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation

under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same

status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,

Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,

Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United

Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: Avenue Marnix 17, B-1000 Brussels

© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 20743:2013: E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST EN ISO 20743:2013
EN ISO 20743:2013 (E)
Contents Page

Foreword ..............................................................................................................................................................3

---------------------- Page: 4 ----------------------
SIST EN ISO 20743:2013
EN ISO 20743:2013 (E)
Foreword

This document (EN ISO 20743:2013) has been prepared by Technical Committee ISO/TC 38 "Textiles" in

collaboration with Technical Committee CEN/TC 248 “Textiles and textile products” the secretariat of which is

held by BSI.

This European Standard shall be given the status of a national standard, either by publication of an identical

text or by endorsement, at the latest by January 2014, and conflicting national standards shall be withdrawn at

the latest by January 2014.

Attention is drawn to the possibility that some of the elements of this document may be the subject of patent

rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.

This document supersedes EN ISO 20743:2007.

According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following

countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech

Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,

Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,

Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.

Endorsement notice

The text of ISO 20743:2013 has been approved by CEN as EN ISO 20743:2013 without any modification.

---------------------- Page: 5 ----------------------
SIST EN ISO 20743:2013
---------------------- Page: 6 ----------------------
SIST EN ISO 20743:2013
INTERNATIONAL ISO
STANDARD 20743
Second edition
2013-07-15
Textiles — Determination of
antibacterial activity of textile products
Textiles — Détermination de l’activité antibactérienne des produits
textiles
Reference number
ISO 20743:2013(E)
ISO 2013
---------------------- Page: 7 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2013

All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form

or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior

written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of

the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2013 – All rights reserved
---------------------- Page: 8 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 Safety precaution ................................................................................................................................................................................................. 2

5 Apparatus ..................................................................................................................................................................................................................... 2

6 Reagents and culture media ..................................................................................................................................................................... 4

6.1 Water ............................................................................................................................................................................................................... 4

6.2 Tryptone soya broth (TSB) ........................................................................................................................................................... 4

6.3 Tryptone soya agar (TSA) .............................................................................................................................................................. 4

6.4 Agar for transfer .................................................................................................................................................................................... 5

6.5 Nutrient broth (NB) ............................................................................................................................................................................ 5

6.6 Peptone salt solution ......................................................................................................................................................................... 5

6.7 Physiological saline ............................................................................................................................................................................. 5

6.8 SCDLP medium ....................................................................................................................................................................................... 6

6.9 Dilution buffer for shake-out bacterial suspension................................................................................................. 6

6.10 Neutralizing solution ......................................................................................................................................................................... 6

6.11 Enumeration agar (EA) .................................................................................................................................................................... 7

6.12 Agar for printing .................................................................................................................................................................................... 7

6.13 Cryoprotective solution for bacterial species ............................................................................................................... 7

6.14 Stock solution of ATP standard reagent ............................................................................................................................ 7

6.15 Buffer solution for ATP luminescent reagent ............................................................................................................... 8

6.16 ATP luminescent reagent ............................................................................................................................................................... 8

6.17 ATP extracting reagent ..................................................................................................................................................................... 8

6.18 ATP eliminating reagent ................................................................................................................................................................. 8

6.19 SCDLP or other medium for preparing ATP reference solution ................................................................... 9

6.20 Shake-out physiological saline .................................................................................................................................................. 9

7 Reference strains ................................................................................................................................................................................................. 9

7.1 Strains ............................................................................................................................................................................................................. 9

7.2 Storage of strains ...............................................................................................................................................................................10

8 Test procedures ..................................................................................................................................................................................................11

8.1 Absorption method (see Annex E) ......................................................................................................................................11

8.2 Transfer method (see Annex E) .............................................................................................................................................15

8.3 Printing method (see Annex E) ..............................................................................................................................................18

9 Test report ................................................................................................................................................................................................................22

Annex A (normative) Strain numbers ..............................................................................................................................................................23

Annex B (normative) Shaking method ............................................................................................................................................................24

Annex C (normative) Quantitative measurement by plate count method ..................................................................25

Annex D (normative) Quantitative measurement by luminescence method ..........................................................26

Annex E (informative) Testing examples .......................................................................................................................................................28

Annex F (informative) Efficacy of antibacterial activity ................................................................................................................31

Bibliography .............................................................................................................................................................................................................................32

© ISO 2013 – All rights reserved iii
---------------------- Page: 9 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
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. 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. 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.
The committee responsible for this document is ISO/TC 38, Textiles.

This second edition cancels and replaces the first edition (ISO 20743:2007), which has been

technically revised.
iv © ISO 2013 – All rights reserved
---------------------- Page: 10 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
Introduction

Speciality products of antibacterial-treated textiles have been introduced in the market and are expanding

year by year in various applications. Those textiles certainly meet the consumer’s requirement to seek

prevention and protection from the negative effects caused by bacteria and to secure the quality of life.

In this situation, the test methods to determine the antibacterial activity for antibacterial textile products

were expected to be established in order to address the substantial need for an International Standard.

The test method for antibacterial activity was developed as ISO 20645 which was a qualitative test

method. There are no testing standards for the quantitative method which gives more objective

information for the antibacterial activity of the textile products.

There are several practical test methods to determine the quantitative antibacterial activity specified

in this International Standard. The test methods are composed of 2 major steps, such as inoculation of

bacteria and quantitative measurement of bacteria.

The methods for the inoculation of bacteria specified in this International Standard are the absorption

method, transfer method and printing method.

The methods of the quantitative measurement of bacteria specified in this International Standard are

colony plate count method and ATP luminescence methods.

Although there are 6 ways for the combination of inoculation methods and quantitative measurements

to execute this test, the choice of the ways depends on the user’s availability and consensus between the

concerned parties.
© ISO 2013 – All rights reserved v
---------------------- Page: 11 ----------------------
SIST EN ISO 20743:2013
---------------------- Page: 12 ----------------------
SIST EN ISO 20743:2013
INTERNATIONAL STANDARD ISO 20743:2013(E)
Textiles — Determination of antibacterial activity of
textile products
1 Scope

This International Standard specifies quantitative test methods to determine the antibacterial activity

of all antibacterial textile products including nonwovens.

This International Standard is applicable to all textile products, including cloth, wadding, thread and

material for clothing, bedclothes, home furnishings and miscellaneous goods, regardless of the type of

antibacterial agent used (organic, inorganic, natural or man-made) or the method of application (built-

in, after-treatment or grafting).

Based on the intended application and on the environment in which the textile product is to be used

and also on the surface properties of the textile properties, the user can select the most suitable of the

following three inoculation methods on determination of antibacterial activity:

a) absorption method (an evaluation method in which the test bacterial suspension is inoculated

directly onto specimens);

b) transfer method (an evaluation method in which test bacteria are placed on an agar plate and

transferred onto specimens);

c) printing method (an evaluation method in which test bacteria are placed on a filter and printed

onto specimens).

The colony plate count method and the ATP (ATP = Adenosine Tri-phosphate) luminescence method are

also specified for measuring the enumeration of bacteria.
2 Normative references

The following referenced documents are indispensable for the application of this document. For dated

references, only the edition cited applies. For undated references, the latest edition of the referenced

document (including any amendments) applies.
ISO 6330, Textiles — Domestic washing and drying procedures for textile testing
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
control fabric

fabric used to validate the growth condition of test bacteria and validate the test

Note 1 to entry: The same fabric as the fabric to be tested but without antibacterial treatment or a 100 % cotton

fabric without fluorescent brighteners or other finish can be used.
3.2
antibacterial agent

product designed to prevent or mitigate the growth of bacteria, to reduce the number of bacteria or to

kill bacteria
© ISO 2013 – All rights reserved 1
---------------------- Page: 13 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
3.3
antibacterial finish

treatment designed to prevent or mitigate the growth of bacteria, to reduce the number of bacteria or

to kill bacteria
3.4
antibacterial activity

activity of an antibacterial finish used to prevent or mitigate the growth of bacteria, to reduce the

number of bacteria or to kill bacteria
3.5
plate count method

method in which the number of bacteria present after incubation is calculated by counting the number

of colonies according to a ten-time dilution method
Note 1 to entry: The results are expressed in “CFU (Colony Forming Unit)”.
3.6
luminescence method
method in which the amount of ATP contained in bacterial cells is measured
Note 1 to entry: The results are expressed in “moles of ATP”.
3.7
neutralizer

chemical agents used to inactivate, neutralize or quench the antibacterial properties of antibacterial agents

4 Safety precaution

The test methods specified in this International Standard require the use of bacteria.

These tests should be carried out by persons with training and experience in the use of microbiological

techniques.

Appropriate safety precautions should be observed with due consideration given to country-specific

regulations.
5 Apparatus
Usual laboratory apparatus and, in particular, the following.

5.1 Spectrophotometer, capable of measuring at a 620 nm to 660 nm wavelength, or McFarland’s

nephelometer.
5.2 Incubator, capable of maintaining a constant temperature of 37 °C ± 2 °C.

5.3 Water baths, one capable of maintaining a constant temperature of 46 °C ± 2 °C and another

capable of maintaining a temperature of 70 °C to 90 °C.
5.4 Mixer, producing a vortex shaking action.

5.5 Stomacher, capable of speeds of 6 blows per second to 8 blows per second, with the corresponding

disposable containers.
5.6 Clean bench, for microbial test.
5.7 Washing machine, in accordance with the specifications of ISO 6330.
2 © ISO 2013 – All rights reserved
---------------------- Page: 14 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)

5.8 Humidity chamber, tropical chamber or other container capable of maintaining a high-humidity

more than 70 %RH atmospheric condition.
−12 −7

5.9 Luminescence photometer, capable of measuring ATP of 10 mol/l to 10 mol/l at 300 nm to

650 nm with a luminescence-measuring reagent.

5.10 Printing apparatus, capable of applying a 4 N load to a test specimen and rotating the specimen

180° in one direction for a period of 3,0 s.

5.11 Refrigerator, capable of maintaining a temperature of between 2 °C and 8 °C.

5.12 Freezers, one adjustable to a temperature below −70 °C and another to a temperature below −20 °C.

5.13 Balance, which can be read to the nearest 0,01 g.

5.14 Filtering apparatus, consisting of an upper container equipped with a membrane filter and a

lower container equipped with a suction opening.

5.15 Pipette, having the most suitable volume for each use, with a tip made of glass or plastic, and with

a tolerance of 0,5 % or less.

5.16 Vials, 30 ml glass bottles, with screw openings, polytetrafluoroethylene or silicone packing and

caps made of polypropylene, polycarbonate or another suitable material.

5.17 Petri dishes, that have been sterilized, made of glass or plastic, in diameter sizes of 90 mm to

100 mm or 55 mm to 60 mm.
5.18 Glass rod, with a diameter of approximately 18 mm.
5.19 Anti-bumping granules (glass beads), with a diameter of 3 mm to 4 mm.
5.20 Erlenmeyer flask, of capacity 100 ml.

5.21 Cutting template, made of a sterilizable material (stainless steel or glass) with a diameter of

3,8 cm ± 0,1 cm.

5.22 Disposable plastic bags, sterile bags suitable for containing food products, to be used for one of

the shaking methods of the specimens.
5.23 Tweezers, made of a material which can be sterilized.

5.24 Stainless-steel cylinder, with a mass of 200 g ± 10 g and a diameter of 3,5 cm ± 0,1 cm.

5.25 Metal wire basket, for autoclaving.
5.26 Aluminium foil.
5.27 Reciprocal incubation shaker.
5.28 Autoclave, capable of sterilizing at 121 °C ± 2 °C and 103 kPa ± 5 kPa.
© ISO 2013 – All rights reserved 3
---------------------- Page: 15 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
6 Reagents and culture media

Reagents used in tests shall be of analytical quality and/or suited for microbiological purposes.

Dehydrated products available on the commercial market are recommended for use in preparing the culture

media. The manufacturer’s instructions for the preparation of these products should be strictly followed.

6.1 Water

Water used in tests shall be analytical-grade water for microbiological media preparation, which is

freshly distilled and/or ion-exchanged and/or ultra-filtered and/or filtered with RO (reverse osmosis).

It shall be free from all toxic or bacteria inhibitory substances.
6.2 Tryptone soya broth (TSB)
Tryptone, pancreatic digest of casein 17 g
Soya peptone, papain digest of soya 3 g
Sodium chloride (NaCl) 5 g
Glucose 2,5 g
Dipotassium hydrogen phosphate 2,5 g
Water 1 000 ml
Mix well and adjust pH, 7,2 ± 0,2
then sterilize by autoclave (5.28).
6.3 Tryptone soya agar (TSA)
Tryptone, pancreatic digest of casein 15 g
Soya peptone, papain digest of soya 5 g
Sodium chloride (NaCl) 5 g
Agar 15 g
Water 1 000 ml
Mix well and adjust pH, 7,2 ± 0,2
then sterilize by autoclave (5.28).
4 © ISO 2013 – All rights reserved
---------------------- Page: 16 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
6.4 Agar for transfer
Tryptone, pancreatic digest of casein 0,75 g
Soya peptone, papain digest of soya 0,25 g
Sodium chloride (NaCl) 5 g
Agar 15 g
Water 1 000 ml
Mix well and adjust pH, 7,2 ± 0,2
then sterilize by autoclave (5.28).
6.5 Nutrient broth (NB)
Beef extract 3 g
Peptone 5 g
Water 1 000 ml
Mix well and adjust pH, then sterilize by
autoclave (5.28).
pH 6,9 ± 0,2
6.6 Peptone salt solution
Peptone, pancreatic digest of 1 g
casein
Sodium chloride (NaCl) 8,5 g
Water 1 000 ml
Mix well and adjust pH, 6,9 ± 0,2
then sterilize by autoclave
(5.28).
6.7 Physiological saline
Sodium chloride (NaCl) 8,5 g
Water 1 000 ml
Mix well, then sterilize by autoclave (5.28).
© ISO 2013 – All rights reserved 5
---------------------- Page: 17 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
6.8 SCDLP medium
Peptone, digest of casein 17 g
Peptone, digest of soybean 3 g
Sodium chloride (NaCl) 5 g
Dipotassium hydrogenphosphate 2,5 g
Glucose 2,5 g
Lecithin 1 g
Polysorbate 80 7 g
Water 1 000 ml
Mix well and adjust pH, 7,2 ± 0,2
then sterilize by autoclave (5.28).

If the neutralizing power is insufficient, the content of polysorbate 80 or lecithin may be adjusted or

another neutralizing agent may be added. The use of any unspecified neutralizer shall be recorded along

with the name and concentration.
6.9 Dilution buffer for shake-out bacterial suspension

This buffer solution consists of 0,005 mol/l sodium dihydrogenphosphate containing 0,037 % sucrose.

pH 7,2 ± 0,2
6.10 Neutralizing solution
The composition of the standard neutralizing solution shall be as follows.
Polysorbate 80 30 g
Egg-yolk lecithin 3 g
Histidine hydrochloride 1 g
Meat or casein peptone 1 g
Sodium chloride (NaCl) 4,3 g
Monopotassium phosphate 3,6 g
Disodium phosphate dihydrate 7,2 g
Water 1 000 ml
Mix well and sterilize by autoclave (5.28).

If the neutralizing power is insufficient, the content of polysorbate 80 or lecithin may be adjusted or

another neutralizing agent may be added. The use of any unspecified neutralizer shall be recorded along

with the name and concentration.
6 © ISO 2013 – All rights reserved
---------------------- Page: 18 ----------------------
SIST EN ISO 20743:2013
ISO 20743:2013(E)
6.11 Enumeration agar (EA)
Dehydrated yeast extract 2,5 g
Casein tryptone 5,0 g
Glucose 1,0 g
Agar 12 g to 18 g (depending on the gel strength of the product)
Water 1 000 ml
Mix well and adjust pH, 7,2 ± 0,2
then sterilize by autoclave (5.28).
6.12 Agar for printing
Agar 20 g
Water 1 000 ml
Mix well and sterilize by autoclave (5.28).
6.13 Cryoprotective solution for bacterial species

For freezing, a cryoprotective solution containing 150 g/l of glycerol or 100 g/l of dimethylsulfoxide

shall be used and prepared as follows,
TSB (6.2) or NB (6.5): 1 000 ml
Add,
Glycerol: 150 g
dimethylsulfoxide: 100 g
Mix well and sterilize by autoclave (5.28).

For solutions containing glycerol, sterilize the mixed solution by autoclave (5.28). For solutions

containing dimethylsulfoxide, sterilize the mixed solution by using 0,22 µm membrane filter.

NOTE Any commercially available product may be used as long as it is a cryoprotective solution or preserving

system that contains glycerol or dimethylsulfoxide and allows preservation of the strains in the same manner as

the specified solutions.
6.14 Stock solution of ATP standard reagent

The concentration of ATP standard reagent is 1 X 10 mol/l which is obtained by the following mixing.

Adenosine-di
...

Questions, Comments and Discussion

Ask us and Technical Secretary will try to provide an answer. You can facilitate discussion about the standard in here.