Cosmetics - Microbiology - Enumeration of yeast and mould (ISO 16212:2017)

This document gives general guidelines for enumeration of yeast and mould present in cosmetics by
counting the colonies on selective agar medium after aerobic incubation.
In order to ensure product quality and safety for consumers, it is advisable to perform an appropriate
microbiological risk analysis to determine the types of cosmetic products to which this document is
applicable. Products considered to present a low microbiological risk (see ISO 29621) include those
with low water activity or extreme pH values, hydro-alcoholic products, etc.
Because of the large variety of cosmetic products within this field of application, this method might not
be suited to some products in every detail (e.g. certain water-immiscible products). Other methods (e.g.
automated) can be substituted for the tests presented here provided that their equivalence has been
demonstrated or the method has been otherwise shown to be suitable.
Yeast enumerated can be identified using suitable identification tests, for example, tests described
in the standards listed in the Bibliography. Mould enumerated can be identified by other appropriate
methods, if necessary.

Kosmetische Mittel - Mikrobiologie - Zählung von Hefen und Schimmelpilzen (ISO 16212:2017)

Cosmétiques - Microbiologie - Dénombrement des levures et des moisissures (ISO 16212:2017)

L'ISO 16212:2017 donne des lignes directrices générales pour le dénombrement des levures et des moisissures présentes dans les cosmétiques par dénombrement des colonies en milieu gélosé sélectif après une incubation aérobie.
Pour garantir la qualité du produit et la sécurité des consommateurs, il est conseillé d'effectuer une analyse de risque microbiologique appropriée, afin de déterminer les types de produits cosmétiques qui relèvent de l'ISO 16212:2017. Les produits considérés comme présentant un faible risque microbiologique (voir l'ISO 29621) comprennent ceux ayant une faible activité de l'eau ou des valeurs de pH extrêmes, les produits hydro-alcooliques, etc.
En raison de la grande variété de produits cosmétiques entrant dans ce domaine d'application, la présente méthode pourrait ne pas être applicable en tout point à certains produits (par exemple à certains produits non miscibles à l'eau). Il est possible de remplacer les essais présentés ici par d'autres méthodes (par exemple des méthodes automatisées) sous réserve que leur équivalence ait été démontrée ou que la méthode ait été par ailleurs indiquée comme adéquate.
Les levures dénombrées peuvent être identifiées à l'aide d'essais d'identification appropriés, par exemple ceux décrits dans les normes indiquées dans la Bibliographie. Les moisissures dénombrées peuvent être identifiées en utilisant d'autres méthodes appropriées, si nécessaire.

Kozmetika - Mikrobiologija - Ugotavljanje števila kvasovk in plesni (ISO 16212:2017)

V tem dokumentu so podane splošne smernice za ugotavljanje števila kvasovk in plesni v kozmetičnih izdelkih s štetjem kolonij na selektivnem agarskem gojišču po aerobni inkubaciji.
Za zagotovitev kakovosti in varnosti izdelkov za stranke je priporočljivo izvesti ustrezno mikrobiološko analizo tveganja, s katero se določijo vrste kozmetičnih izdelkov, za katere se uporablja ta dokument. Izdelki, ki po ocenah predstavljajo nizko mikrobiološko tveganje (glej ISO 29621), vključujejo izdelke z nizko aktivnostjo vode ali s skrajnimi vrednostmi pH, hidro-alkoholne izdelke itd.
Zaradi velike raznolikosti kozmetičnih izdelkov na tem področju uporabe ta metoda morda ni primerna za nekatere izdelke (npr. tiste, ki se ne mešajo z vodo). Druge metode (npr. avtomatizirane) lahko nadomestijo v tem dokumentu opisani preskusi, če je bila dokazana njihova enakovrednost ali je bila metoda kako drugače opredeljena kot primerna.
Naštete kvasovke je mogoče identificirati z ustreznimi identifikacijskimi preskusi, na primer s preskusi, opisanimi v standardih, navedenih v bibliografiji. Naštete plesni je mogoče identificirati z drugimi ustreznimi metodami, če je to potrebno.

General Information

Status
Published
Public Enquiry End Date
07-Mar-2017
Publication Date
08-Oct-2017
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
06-Sep-2017
Due Date
11-Nov-2017
Completion Date
09-Oct-2017

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SLOVENSKI STANDARD
SIST EN ISO 16212:2017
01-november-2017
1DGRPHãþD
SIST EN ISO 16212:2011
Kozmetika - Mikrobiologija - Ugotavljanje števila kvasovk in plesni (ISO
16212:2017)
Cosmetics - Microbiology - Enumeration of yeast and mould (ISO 16212:2017)
Kosmetische Mittel - Mikrobiologie - Zählung von Hefen und Schimmelpilzen (ISO
16212:2017)
Cosmétiques - Microbiologie - Dénombrement des levures et des moisissures (ISO
16212:2017)
Ta slovenski standard je istoveten z: EN ISO 16212:2017
ICS:
07.100.40 Kozmetika - mikrobiologija Cosmetics microbiology
SIST EN ISO 16212:2017 en,fr,de

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

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SIST EN ISO 16212:2017
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SIST EN ISO 16212:2017
EN ISO 16212
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2017
EUROPÄISCHE NORM
ICS 07.100.40 Supersedes EN ISO 16212:2011
English Version
Cosmetics - Microbiology - Enumeration of yeast and
mould (ISO 16212:2017)

Cosmétiques - Microbiologie - Dénombrement des Kosmetische Mittel - Mikrobiologie - Zählung von

levures et des moisissures (ISO 16212:2017) Hefen und Schimmelpilzen (ISO 16212:2017)

This European Standard was approved by CEN on 26 April 2017.

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, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

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

© 2017 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16212:2017 E

worldwide for CEN national Members.
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SIST EN ISO 16212:2017
EN ISO 16212:2017 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

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SIST EN ISO 16212:2017
EN ISO 16212:2017 (E)
European foreword

This document (EN ISO 16212:2017) has been prepared by Technical Committee ISO/TC 217

"Cosmetics" in collaboration with Technical Committee CEN/TC 392 “Cosmetics” the secretariat of

which is held by AFNOR.

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 2018 and conflicting national standards shall

be withdrawn at the latest by January 2018.

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

patent rights. CEN shall not be held responsible for identifying any or all such patent rights.

This document supersedes EN ISO 16212:2011.

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, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

Turkey and the United Kingdom.
Endorsement notice

The text of ISO 16212:2017 has been approved by CEN as EN ISO 16212:2017 without any modification.

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SIST EN ISO 16212:2017
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SIST EN ISO 16212:2017
INTERNATIONAL ISO
STANDARD 16212
Second edition
2017-06
Cosmetics — Microbiology —
Enumeration of yeast and mould
Cosmétiques — Microbiologie — Dénombrement des levures et des
moisissures
Reference number
ISO 16212:2017(E)
ISO 2017
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SIST EN ISO 16212:2017
ISO 16212:2017(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2017, Published in Switzerland

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
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2017 – All rights reserved
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SIST EN ISO 16212:2017
ISO 16212:2017(E)
Contents Page

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

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

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

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

4 Principles ..................................................................................................................................................................................................................... 2

4.1 General ........................................................................................................................................................................................................... 2

4.2 Plate count .................................................................................................................................................................................................. 2

4.3 Membrane filtration ........................................................................................................................................................................... 2

5 Diluents, neutralizers and culture media ................................................................................................................................... 3

5.1 General ........................................................................................................................................................................................................... 3

5.2 Neutralizing diluents and diluents ........................................................................................................................................ 3

5.3 Diluent for yeast suspension (tryptone sodium chloride solution) .......................................................... 4

5.4 Culture media ........................................................................................................................................................................................... 4

6 Apparatus and glassware ............................................................................................................................................................................ 5

7 Strain of microorganisms ............................................................................................................................................................................ 5

8 Handling of cosmetic products and laboratory samples ............................................................................................ 6

9 Procedure..................................................................................................................................................................................................................... 6

9.1 General recommendation .............................................................................................................................................................. 6

9.2 Preparation of the initial suspension .................................................................................................................................. 6

9.2.1 General...................................................................................................................................................................................... 6

9.2.2 Water-miscible products........................................................................................................................................... 6

9.2.3 Water-immiscible products .................................................................................................................................... 6

9.3 Counting methods ................................................................................................................................................................................ 6

9.3.1 Dilutions for counting methods .......................................................................................................................... 6

9.3.2 Plate-count methods .................................................................................................................................................... 7

10 Counting of colonies (plate counts and membrane filtration methods) ....................................................7

11 Expression of results ........................................................................................................................................................................................ 8

11.1 Method of calculation for plate count ................................................................................................................................. 8

11.2 Interpretation .......................................................................................................................................................................................... 9

12 Neutralization of the antifungicidal properties of the product ........................................................................10

12.1 General ........................................................................................................................................................................................................10

12.2 Preparation of inoculum ..............................................................................................................................................................11

12.3 Suitability of counting methods ............................................................................................................................................11

12.3.1 Principle ...............................................................................................................................................................................11

12.3.2 Suitability test of the pour-plate method ................................................................................................11

12.3.3 Suitability of the surface spread method .................................................................................................11

12.3.4 Suitability of the membrane filtration method ...................................................................................11

13 Test report ................................................................................................................................................................................................................12

Annex A (informative) Other neutralizing diluents ...........................................................................................................................13

Annex B (informative) Other diluents ..............................................................................................................................................................15

Annex C (informative) Other culture media ...............................................................................................................................................16

Annex D (informative) Neutralizers of antifungicidal activity of preservatives and

rinsing liquids ......................................................................................................................................................................................................18

Bibliography .............................................................................................................................................................................................................................19

© ISO 2017 – All rights reserved iii
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SIST EN ISO 16212:2017
ISO 16212:2017(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 (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 voluntary nature of standards, 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: w w w . i s o .org/ iso/ foreword .html.
This document was prepared by Technical Committee ISO/TC 217, Cosmetics.

This second edition cancels and replaces the first edition (ISO 16212:2008), of which it constitutes a

minor revision. The changes compared to the previous edition are as follows:

— in the Scope, “see ISO 29621” has been added and the reference has been added to the Bibliography;

— in the Scope, “used” has been changed to “substituted” and “validated” has been changed to “shown

to be suitable”;
— in 4.1, “validated” has been changed to “demonstrated”;

— in 4.3, “by a valid method” has been changed to “as described in Clause 12” and “validated procedure”

has been replaced by “described procedure”;
— in 5.1, “specifications” has been changed to “instructions”;
— in 5.2.3.1.2, “peptone” has been changed to “peptic digest of animal tissue”;
— in Clause 7, “validation” has been changed to “suitability”;
— in 9.3.2.1, “validated” has been changed to “demonstrated to be suitable”;

— in 9.3.2.3, “prepared as validated” has been changed to “demonstrated to be suitable”;

— in 11.2.1, “validated according to” has been changed to “demonstrated to be suitable for”;

— in 12.3, “validation” has been changed to “suitability”;

— in 12.3.2, instances of “validation” have been changed to “suitability test” and “validated” has been

changed to “satisfactory”;

— in 12.3.3, the first instance of “validation” has been changed to “suitability” and the second instance

has been changed to “suitability test”; “validated” has been changed to “satisfactory”;

iv © ISO 2017 – All rights reserved
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SIST EN ISO 16212:2017
ISO 16212:2017(E)

— in 12.3.4, the first instance of “validation” has been changed to “suitability” and the second instance

has been changed to “suitability test”; “validated” has been changed to “satisfactory”;

— in Clause 13 f), “validation” has been changed to “suitability”;

— in A.1, B.1 and C.1, “validated” has been changed to “demonstrated to be suitable”.

© ISO 2017 – All rights reserved v
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SIST EN ISO 16212:2017
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SIST EN ISO 16212:2017
INTERNATIONAL STANDARD ISO 16212:2017(E)
Cosmetics — Microbiology — Enumeration of yeast and
mould
1 Scope

This document gives general guidelines for enumeration of yeast and mould present in cosmetics by

counting the colonies on selective agar medium after aerobic incubation.

In order to ensure product quality and safety for consumers, it is advisable to perform an appropriate

microbiological risk analysis to determine the types of cosmetic products to which this document is

applicable. Products considered to present a low microbiological risk (see ISO 29621) include those

with low water activity or extreme pH values, hydro-alcoholic products, etc.

Because of the large variety of cosmetic products within this field of application, this method might not

be suited to some products in every detail (e.g. certain water-immiscible products). Other methods (e.g.

automated) can be substituted for the tests presented here provided that their equivalence has been

demonstrated or the method has been otherwise shown to be suitable.

Yeast enumerated can be identified using suitable identification tests, for example, tests described

in the standards listed in the Bibliography. Mould enumerated can be identified by other appropriate

methods, if necessary.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements 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 21148, Cosmetics — Microbiology — General instructions for microbiological examination

EN 12353, Chemical disinfectants and antiseptics — Preservation of test organisms used for the

determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal and virucidal

(including bacteriophages) activity
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at http:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
3.1
yeast

single-cell fungus, which multiplies mainly vegetatively by budding, able to grow under the test

conditions specified in this document
3.2
mould

mycelium forming microfungus, including spores and conidia, able to grow under the test conditions

specified in this document
© ISO 2017 – All rights reserved 1
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SIST EN ISO 16212:2017
ISO 16212:2017(E)
3.3
product
portion of an identified cosmetic product received in the laboratory for testing
3.4
sample

portion of the product (3.3) (at least 1 g or 1 ml) that is used in the test to prepare the initial suspension

3.5
initial suspension

suspension (or solution) of the sample (3.4) in a defined volume of an appropriate enrichment broth

3.6
sample dilution
dilution of the initial suspension (3.5)
4 Principles
4.1 General

This method involves enumeration of colonies on a selective agar medium. The possible inhibition of

[5]

fungal growth by the sample shall be neutralized to allow the detection of viable microorganisms .

In all cases and whatever the methodology, the neutralization of the antifungicidal properties of the

[6][8][9]
product shall be checked and demonstrated .
4.2 Plate count
Plate count consists of the following steps.

— Preparation of poured plates, or spread plates, using a specified culture medium, and inoculation of

the plates using a defined quantity of the initial suspension or dilution of the product.

— Aerobic incubation of the plates at 25 °C ± 2,5 °C for 3 d to 5 d.

— Counting of the number of colony-forming units (CFU) and calculation of the amount of yeast and

mould per millilitre or per gram of product.

NOTE An alternative condition for incubation is 22,5 °C ± 2,5 °C for 5 d to 7 d using the culture medium

without antibiotic.
4.3 Membrane filtration
Membrane filtration consists of the following steps.

— Transfer a suitable amount of the sample, prepared as described in Clause 12, in the filtration

apparatus, wetted with a small volume of an appropriate sterile diluent. Filter immediately and

wash according to the described procedure (see 12.3.4). Transfer the membrane filter onto the

surface of the specified agar medium as specified in ISO 21148.
— Aerobic incubation of the membranes at 25 °C ± 2,5 °C for 3 d to 5 d.

— Counting of the number of colony-forming units (CFU) and calculation of the amount of yeast and

mould per millilitre or per gram of product.

NOTE An alternative condition for incubation is 22,5 °C ± 2,5 °C for 5 d to 7 d using the culture medium

without antibiotic.
2 © ISO 2017 – All rights reserved
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SIST EN ISO 16212:2017
ISO 16212:2017(E)
5 Diluents, neutralizers and culture media
5.1 General

General instructions are given in ISO 21148. When water is mentioned in this document, use distilled

water or purified water as specified in ISO 21148.

The following diluents, neutralizers and culture media are suitable for enumeration of yeasts and

moulds. Other diluents, neutralizers and culture media may be used if they have been demonstrated to

be suitable for use.
5.2 Neutralizing diluents and diluents
5.2.1 General

The diluent is used to disperse the sample. It may contain neutralizers if the sample to be tested

has antifungicidal properties. The efficacy of the neutralization shall be demonstrated before the

determination of the count (see Clause 12). Information relative to suitable neutralizers is given in

Annex D.
5.2.2 Neutralizing diluent
5.2.2.1 Fluid casein digest–soy lecithin–polysorbate 20 medium (SCDLP 20 broth)
5.2.2.1.1 Composition
Pancreatic digest of casein 20,0 g
Soy lecithin 5,0 g
Polysorbate 20 40 ml
Water 960 ml
5.2.2.1.2 Preparation

Dissolve the polysorbate 20 in 960 ml of water by mixing while heating in a water bath at 49 °C ± 2 °C.

Add pancreatic digest of casein and soy lecithin. Heat for about 30 min to effect solution. Mix and

dispense the medium into suitable containers. Sterilize in the autoclave at 121 °C for 15 min. After

sterilization, the pH shall be equivalent to 7,3 ± 0,2 when measured at room temperature.

5.2.2.2 Other neutralizing diluents

Other neutralizing diluents may be used as appropriate (see Annex A and Annex D).

5.2.3 Diluent
5.2.3.1 Fluid A
5.2.3.1.1 Composition
Peptic digest of animal tissue 1,0 g
Water 1 000 ml
© ISO 2017 – All rights reserved 3
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SIST EN ISO 16212:2017
ISO 16212:2017(E)
5.2.3.1.2 Preparation

Dissolve 1 g of peptic digest of animal tissue in water to make 1 l. Heat with frequent agitation. Dispense

into suitable containers. Sterilize in the autoclave at 121 °C for 15 min. After sterilization, the pH shall

be equivalent to 7,1 ± 0,2 when measured at room temperature.
5.2.3.2 Other diluents
Other diluents may be used as appropriate (see Annex B).
5.3 Diluent for yeast suspension (tryptone sodium chloride solution)
5.3.1 Composition
Tryptone, pancreatic digest of casein 1,00 g
Sodium chloride 8,50 g
Water 1 000 ml
5.3.2 Preparation

Dissolve the components in the water by mixing while heating. Dispense into suitable containers.

Sterilize in the autoclave at 121 °C for 15 min. After sterilization, the pH shall be equivalent to 7,0 ± 0,2

when measured at room temperature.
5.4 Culture media
5.4.1 General

Culture media may be prepared as follows or from dehydrated culture media according to the

manufacturer’s instructions. Ready-to-use media may be used when their composition and/or growth

yields are comparable to those of the formulae given herein.
5.4.2 Sabouraud dextrose chloramphenicol agar medium (SDCA)
5.4.2.1 Composition
Dextrose 40,0 g
Peptic digest of animal tissue 5,0 g
Pancreatic digest of casein 5,0 g
Chloramphenicol 0,050 g
Agar 15,0 g
Water 1 000 ml
5.4.2.2 Preparation

Dissolve the components (including the chloramphenicol) or the dehydrated complete medium in the

water by mixing while heating. Dispense the medium into suitable containers. Sterilize in an autoclave

4 © ISO 2017 – All rights reserved
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SIST EN ISO 16212:2017
ISO 16212:2017(E)

at 121 °C for 15 min. After sterilization, the pH shall be equivalent to 5,6 ± 0,2 when measured at room

temperature.

NOTE For known and non-contaminated products (with bacteria), the media are used without

chloramphenicol.
5.4.3 Other media
Other media may be used as appropriate (see Annex C).

5.4.4 Agar medium for cultivation of reference strain: Sabouraud dextrose agar medium (SDA)

5.4.4.1 Composition
Dextrose 40,0 g
Peptic digest of animal tissue 5,0 g
Pancreatic digest of casein 5,0 g
Agar 15,0 g
Water 1 000 ml
5.4.4.2 Preparation

Dissolve the components or the dehydrated complete medium in the water by mixing while heating.

Dispense the medium into suitable containers. Sterilize in an autoclave at 121 °C for 15 min. After

sterilization, the pH shall be equivalent to 5,6 ± 0,2 when measured at room temperature.

6 Apparatus and glassware
The laboratory equipment, apparatus and glassware are described in ISO 21148.
7 Strain of microorganisms
For testing the efficacy of neutralizers, one yeast reference strain is used:
1) 2) 3) 4)

— Candida albicans ATCC 10231 or equivalent strain (IP 48.72 or NCPF 3179 or NBRC 1594 or

5) 6)
KCTC 17205 or TISTR 5779) or other equivalent national collection strain.

The selected yeast strain being considered more susceptible to antifungicidal activity than moulds

may be accepted as representative of fungi (yeast and mould) for the suitability of the methodology.

However, in case of specific needs, the test for the efficacy of neutralizers may be performed with an

additional mould reference strain, using a suitable protocol for the preparation of a calibrated inoculum

[3]
(e.g. see EN 13624:2013, 5.4.1.4 ).

The culture should be reconstituted according to the procedures provided by the supplier of the

reference strain.
1) ATCC = American Type Culture Collection.
2) IP = Institut Pasteur.
3) NCPF = National Collection of Pathogenic Fungi.
4) NBRC = Biological Resource Center, NITE.
5) KCTC = Korean Collection for Type Cultures.
6) TISTR = Thailand Institute of Scientific and Technological Research.
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SIST EN ISO 16212:2017
ISO 16212:2017(E)
The strain may be kept in the laboratory in accordance with EN 12353.
8 Handling of cosmetic products and laboratory samples
If necessary, store products to be tested at room temperature.

Do not incubate, refrigerate or freeze products (3.3) and samples (3.4) before or after analysis.

Sampling of cosmetic products to be analysed should be carried out as described in ISO 21148. Analyse

samples as described in ISO 21148 and according to the procedure described in Clause 9.

9 Procedure
9.1 General recommendation

Use sterile material, equipment and aseptic techniques to prepare the sample, initial suspension and

dilutions. In the case of the preparation of the initial suspension, the time that elapses between the end

of the preparation and the moment the inoculum comes into contact with the culture medium shall not

exceed 45 min, unless specifically mentioned in the established protocols or documents.

9.2 Preparation of
...

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