EN ISO 16212:2017

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

ISO 16212:2017 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 ISO 16212:2017 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
• Publication Date: 04-Jul-2017
• Withdrawal Date: 30-Jan-2018
• ICS: 07.100.40 - Cosmetics microbiology
• ICS: 07.100.99 - Other standards related to microbiology
• ICS: 71.100.70 - Cosmetics. Toiletries
• Technical Committee: CEN/TC 392 - Cosmetics
• Drafting Committee: CEN/TC 392 - Cosmetics
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 05-Jul-2017
• Completion Date: 05-Jul-2017
• Ref Project: SIST EN ISO 16212:2017 - Cosmetics - Microbiology - Enumeration of yeast and mould (ISO 16212:2017)

Relations

• Replaces: EN ISO 16212:2011 - Cosmetics - Microbiology - Enumeration of yeast and mould (ISO 16212:2008)
• Effective Date: 31-Aug-2016
• Amended By: EN ISO 16212:2017/A1:2022 - Cosmetics - Microbiology - Enumeration of yeast and mould - Amendment 1 (ISO 16212:2017/Amd 1:2022)
• Effective Date: 22-Sep-2021

Overview

EN ISO 16212:2017 - Cosmetics - Microbiology - Enumeration of yeast and mould provides general guidelines for the enumeration of yeasts and moulds in cosmetic products. The method is based on counting colony-forming units on selective agar media after aerobic incubation. The document is published by CEN as the European adoption of ISO 16212:2017 and supersedes the 2011 edition.

Key topics and requirements

• Scope and applicability: Guidance for a broad range of cosmetics; users should perform a microbiological risk analysis (see ISO 29621) to determine applicability - some low-risk products (e.g., low water activity, extreme pH, hydro‑alcoholic formulations) may require different approaches.
• Enumeration methods: Describes principal methods such as plate-count (pour or surface spread) and membrane filtration for enumerating yeast and mould colonies.
• Sample handling and preparation: Procedures for preparing initial suspensions of water‑miscible and water‑immiscible products, and general laboratory handling.
• Diluents, neutralizers and culture media: Requirements and options for neutralizing antifungicidal activity, choice of diluents and culture media, with informative annexes listing alternative reagents.
• Suitability tests: Tests to demonstrate that a chosen counting method is suitable for a particular product matrix (e.g., neutralization of preservatives, method equivalence).
• Counting and result expression: Procedures for colony counting, calculation of results, interpretation and reporting requirements.
• Identification: Yeast colonies can be identified using suitable tests referenced in the bibliography; mould identification by appropriate methods when necessary.
• Documentation: Required elements for the test report and informative annexes (diluents, media, neutralizers).

Practical applications and users

• Quality control laboratories in cosmetic manufacturing for routine microbial monitoring and batch release.
• Contract testing labs performing product testing and stability studies.
• Regulatory compliance teams and auditors using standardized methods to evaluate product safety and documentation.
• R&D and formulation scientists validating manufacturing processes or alternative analytical methods (automated methods can be substituted if demonstrated equivalent or shown suitable).
• Microbiology professionals conducting contamination investigations, shelf‑life assessments or preservative effect evaluations (note: neutralization procedures are addressed).

Related standards

• ISO 29621 - Guidance on microbiological risk analysis for cosmetics (referenced in the scope).
• ISO 21148 - (Referenced normative documents for related methods and requirements).
• EN ISO 16212:2017 aligns with ISO/TC 217 (Cosmetics) recommendations and should be used together with product‑specific risk assessments and validated laboratory practices.

Keywords: EN ISO 16212:2017, cosmetics microbiology, enumeration of yeast and mould, selective agar, plate count, membrane filtration, microbiological risk analysis.

Standards Content (Sample)

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

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.

Contents Page
European foreword . 3

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.
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
ISO 16212:2017(E)
© 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. [phone]
Fax [phone]
[e-mail]
www.iso.org
ii © ISO 2017 – All rights reserved

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 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

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”.
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 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

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 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

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 glass
...