Cosmetics - Microbiology - Detection of specified and non-specified microorganisms (ISO 18415:2017)

This document gives general guidelines for the detection and identification of specified microorganisms
in cosmetic products as well as for the detection and identification of other kinds of aerobic mesophilic
non-specified microorganisms in cosmetic products.
Microorganisms considered as specified in this document might differ from country to country
according to national practices or regulations. Most of them considered as specified microorganisms
include one or more of the following species: Pseudomonas aeruginosa, Escherichia coli, Staphylococcus
aureus and Candida albicans.
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, hydro-alcoholic products, extreme pH values, etc.
The method described in this document is based on the detection of microbial growth in a non-selective
liquid medium (enrichment broth) suitable to detect microbial contamination, followed by isolation of
microorganisms on non-selective agar media. Other methods can be appropriate depending on the level
of detection required.
In this document specific indications are given for identification of Pseudomonas aeruginosa, Escherichia
coli, Staphylococcus aureus and Candida albicans. Other microorganisms that grow under the conditions
described in this document may be identified by using suitable tests according to a general scheme (see
Annex A). Other standards (e.g. ISO 18416, ISO 21150, ISO 22717, ISO 22718) may be appropriate.
Because of the large variety of cosmetic products within this field of application, this method might not
be suited in every detail to some products (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.

Kosmetische Mittel - Mikrobiologie - Nachweis von spezifizierten und nichtspezifizierten Mikroorganismen (ISO 18415:2017)

Cosmétiques - Microbiologie - Détection des micro-organismes spécifiés et non spécifiés (ISO 18415:2017)

ISO 18416:2017 donne des lignes directrices générales pour la détection et l'identification de micro-organismes spécifiés dans les produits cosmétiques, et aussi pour la détection et l'identification d'autres sortes de micro-organismes mésophiles aérobies non spécifiés, dans les produits cosmétiques.
Les micro-organismes considérés comme spécifiés dans le présent document peuvent différer d'un pays à l'autre, suivant les pratiques ou réglementations nationales. La plupart des micro-organismes considérés comme spécifiés comprennent une ou plusieurs espèces, parmi les suivantes: Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus et Candida albicans.
Pour garantir la qualité du produit et la sécurité des consommateurs, il est conseillé d'effectuer une analyse appropriée du risque microbiologique afin de déterminer les types de produits cosmétiques qui relèvent du présent document. Les produits considérés comme présentant un faible risque microbiologique comprennent ceux ayant une faible activité de l'eau, les produits hydro-alcooliques, les produits ayant des valeurs de pH extrêmes, etc.
La méthode décrite dans le présent document est fondée sur la détection d'une croissance microbienne dans un milieu liquide non sélectif (bouillon d'enrichissement) qui permet de détecter une contamination microbienne, suivie d'un isolement des micro-organismes sur un milieu gélosé non sélectif. D'autres méthodes peuvent être appropriées, en fonction du niveau de détection exigé.
Dans le présent document, des indications spécifiques sont données pour l'identification de Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus et Candida albicans. D'autres micro-organismes qui se développent dans les conditions décrites dans le présent document peuvent être identifiés en mettant en ?uvre des essais appropriés conformes à un schéma général (voir Annexe A). D'autres Normes internationales (par exemple, l'ISO 18416, l'ISO 21150, l'ISO 22717 et l'ISO 22718) peuvent convenir.
En raison de la grande variété de produits cosmétiques relevant du présent domaine d'application, la présente méthode pourrait ne pas être, en tous points, applicable à certains produits (par exemple certains produits non miscibles à l'eau). D'autres méthodes (par exemple automatisées) peuvent se substituer aux essais présentés ici, sous réserve que leur équivalence ait été démontrée ou que la méthode ait été validée par ailleurs.

Kozmetika - Mikrobiologija - Ugotavljanje prisotnosti specifičnih in nespecifičnih mikroorganizmov (ISO 18415:2017)

Ta dokument podaja splošne smernice za ugotavljanje prisotnosti in identifikacijo specifičnih mikroorganizmov v kozmetičnih izdelkih ter ugotavljanje prisotnosti in identifikacijo drugih vrst aerobnih mezofilnih nespecifičnih mikroorganizmov v kozmetičnih izdelkih.
Mikroorganizmi, ki so v tem dokumentu obravnavani kot specifični, se lahko v različnih državah zaradi državnih praks ali predpisov razlikujejo. Večina od teh, ki so obravnavani kot specifični, vključuje eno ali več naslednjih vrst: Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus in Candida albicans.
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.
Metoda, opisana v tem dokumentu, temelji na ugotavljanju prisotnosti rasti mikrobov v neselektivnem tekočem gojišču (obogatitven bujon), primernem za ugotavljanje mikrobnega onesnaženja, ki mu sledi izolacija mikroorganizmov na neselektivnem agarskem gojišču. Ustrezne so lahko tudi druge metode, odvisno od zahtevane ravni ugotavljanja prisotnosti.
V tem dokumentu so podane specifične navedbe za identifikacijo vrst Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus in Candida albicans. Prisotnost drugih mikroorganizmov, ki rastejo pod pogoji, opisanimi v tem dokumentu, je mogoče ugotoviti z ustreznimi preskusi v skladu s splošno shemo (glej dodatek A). Primerni so lahko drugi standardi (npr. ISO 18416, ISO 21150, ISO 22717, ISO 22718).
Zaradi velike raznolikosti kozmetičnih izdelkov na tem področju uporabe ta metoda morda ni v celoti 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

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

RELATIONS

Buy Standard

Standard
SIST EN ISO 18415:2017 - natisnjeno
English language
28 pages
sale 10% off
Preview
sale 10% off
Preview

e-Library read for
1 day

Standards Content (sample)

SLOVENSKI STANDARD
SIST EN ISO 18415:2017
01-november-2017
1DGRPHãþD
SIST EN ISO 18415:2011

.R]PHWLND0LNURELRORJLMD8JRWDYOMDQMHSULVRWQRVWLVSHFLILþQLKLQQHVSHFLILþQLK

PLNURRUJDQL]PRY ,62

Cosmetics - Microbiology - Detection of specified and non-specified microorganisms

(ISO 18415:2017)

Kosmetische Mittel - Mikrobiologie - Nachweis von spezifizierten und nichtspezifizierten

Mikroorganismen (ISO 18415:2017)

Cosmétiques - Microbiologie - Détection des micro-organismes spécifiés et non spécifiés

(ISO 18415:2017)
Ta slovenski standard je istoveten z: EN ISO 18415:2017
ICS:
07.100.40 Kozmetika - mikrobiologija Cosmetics microbiology
SIST EN ISO 18415:2017 en,fr,de

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

---------------------- Page: 1 ----------------------
SIST EN ISO 18415:2017
---------------------- Page: 2 ----------------------
SIST EN ISO 18415:2017
EN ISO 18415
EUROPEAN STANDARD
NORME EUROPÉENNE
June 2017
EUROPÄISCHE NORM
ICS 07.100.40 Supersedes EN ISO 18415:2011
English Version
Cosmetics - Microbiology - Detection of specified and non-
specified microorganisms (ISO 18415:2017)

Cosmétiques - Microbiologie - Détection des micro- Kosmetische Mittel - Mikrobiologie - Nachweis von

organismes spécifiés et non spécifiés (ISO 18415:2017) spezifizierten und nichtspezifizierten

Mikroorganismen (ISO 18415:2017)
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 18415:2017 E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST EN ISO 18415:2017
EN ISO 18415:2017 (E)
Contents Page

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

---------------------- Page: 4 ----------------------
SIST EN ISO 18415:2017
EN ISO 18415:2017 (E)
European foreword

This document (EN ISO 18415:2017) 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 December 2017 and conflicting national standards

shall be withdrawn at the latest by December 2017.

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 18415:2011.

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 18415:2017 has been approved by CEN as EN ISO 18415:2017 without any modification.

---------------------- Page: 5 ----------------------
SIST EN ISO 18415:2017
---------------------- Page: 6 ----------------------
SIST EN ISO 18415:2017
INTERNATIONAL ISO
STANDARD 18415
Second edition
2017-06
Cosmetics — Microbiology —
Detection of specified and non-
specified microorganisms
Cosmétiques — Microbiologie — Détection des micro-organismes
spécifiés et non spécifiés
Reference number
ISO 18415:2017(E)
ISO 2017
---------------------- Page: 7 ----------------------
SIST EN ISO 18415:2017
ISO 18415:2017(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
---------------------- Page: 8 ----------------------
SIST EN ISO 18415:2017
ISO 18415:2017(E)
Contents Page

Foreword ..........................................................................................................................................................................................................................................v

Introduction ................................................................................................................................................................................................................................vi

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

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

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

4 Principle ........................................................................................................................................................................................................................ 3

5 Diluents and culture media ....................................................................................................................................................................... 3

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

5.2 Diluent for the microbial suspension (tryptone sodium chloride solution) ..................................... 3

5.2.1 General...................................................................................................................................................................................... 3

5.2.2 Composition ......................................................................................................................................................................... 4

5.2.3 Preparation ........................................................................................................................................................................... 4

5.3 Culture media ........................................................................................................................................................................................... 4

5.3.1 General...................................................................................................................................................................................... 4

5.3.2 Enrichment broth ............................................................................................................................................................ 4

5.3.3 Non-selective agar medium .................................................................................................................................... 5

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

7 Strains of microorganism ............................................................................................................................................................................ 6

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

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

9.1 General recommendations............................................................................................................................................................ 6

9.2 Preparation of the initial suspension in the enrichment broth .................................................................... 6

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

9.2.2 Water-miscible products........................................................................................................................................... 7

9.2.3 Water-immiscible products .................................................................................................................................... 7

9.2.4 Filterable products ......................................................................................................................................................... 7

9.3 Incubation of the initial suspension ..................................................................................................................................... 7

9.4 Isolation of specified and non-specified microorganisms ................................................................................. 7

9.5 Procedure for identification of the specified microorganism: Pseudomonas aeruginosa ....... 7

9.5.1 Gram staining ...................................................................................................................................................................... 7

9.5.2 Oxidase test .......................................................................................................................................................................... 7

9.5.3 Identification test ............................................................................................................................................................ 8

9.6 Procedure for identification of the specified microorganism: Escherichia coli ............................... 8

9.6.1 Gram staining ...................................................................................................................................................................... 8

9.6.2 Oxidase test .......................................................................................................................................................................... 8

9.6.3 Identification test ............................................................................................................................................................ 8

9.7 Procedure for identification of the specified microorganism: Staphylococcus aureus ............. 8

9.7.1 Gram staining ...................................................................................................................................................................... 8

9.7.2 Catalase test ......................................................................................................................................................................... 8

9.7.3 Identification test ............................................................................................................................................................ 8

9.8 Procedure for the identification of the specified microorganism: Candida albicans ................. 9

9.8.1 Gram staining ...................................................................................................................................................................... 9

9.8.2 Identification test ............................................................................................................................................................ 9

9.9 Procedure for the identification of non-specified microorganisms .......................................................... 9

9.9.1 Gram staining ...................................................................................................................................................................... 9

9.9.2 Oxidase test .......................................................................................................................................................................... 9

9.9.3 Catalase test ......................................................................................................................................................................... 9

9.9.4 Identification test ............................................................................................................................................................ 9

10 Expression of the results ...........................................................................................................................................................................10

10.1 Detection of specified microorganisms ..........................................................................................................................10

© ISO 2017 – All rights reserved iii
---------------------- Page: 9 ----------------------
SIST EN ISO 18415:2017
ISO 18415:2017(E)

10.2 Detection of non-specified microorganisms...............................................................................................................10

10.3 Absence of microorganisms ......................................................................................................................................................10

11 Neutralization of the antimicrobial properties of the product .........................................................................10

11.1 General ........................................................................................................................................................................................................10

11.2 Preparation of inoculum ..............................................................................................................................................................10

11.3 Suitability of detection method by enrichment .......................................................................................................10

11.3.1 Principle ...............................................................................................................................................................................10

11.3.2 Procedure ............................................................................................................................................................................11

11.3.3 Interpretation of suitability test results ...................................................................................................11

12 Test report ................................................................................................................................................................................................................11

Annex A (informative) General scheme for identification of microorganisms......................................................13

Annex B (informative) Other media ...................................................................................................................................................................14

Annex C (informative) Neutralizers of antimicrobial activity of preservatives and rinsing liquids 17

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

iv © ISO 2017 – All rights reserved
---------------------- Page: 10 ----------------------
SIST EN ISO 18415:2017
ISO 18415:2017(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 18415:2007), of which it constitutes a

minor revision with the following changes:

— 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 3.8, the term “validated” has been changed to “demonstrated to be suitable”;

— in Clause 4, the term “validated” has been changed to “demonstrated”;
— in 5.1, “specifications” has been changed to “instructions”;

— in 5.1, the phrase “are validated” has been changed to “have been demonstrated to be suitable”;

— in 5.2.1, 5.3.3.1, 11.3.1, 11.3.2, instances of the term “validation” and in the heading title of 11.3.3

have been changed to “suitability test”;

— in 11.3, the term “validation” in the heading title has been changed to “suitability”;

— in 11.3.3, instances of “validated” have been changed to “satisfactory”;

— in Clause 12 f), the term “validation” has been changed to “demonstration of the suitability”.

© ISO 2017 – All rights reserved v
---------------------- Page: 11 ----------------------
SIST EN ISO 18415:2017
ISO 18415:2017(E)
Introduction

Microbiological examinations of cosmetic products are carried out according to an appropriate

microbiological risk analysis in order to ensure their quality and safety for consumers.

Microbiological risk analysis depends on several parameters such as:
— potential alteration of cosmetic products;
— pathogenicity of microorganisms;

— site of application of the cosmetic product (hair, skin, eyes, mucous membranes);

— type of user (adults, children including under 3 years).

For cosmetics and other topical products, the detection of skin pathogens such as Staphylococcus

aureus, Pseudomonas aeruginosa and Candida albicans may be relevant because they can cause skin

or eye infection. The detection of other kinds of microorganisms might be of interest since these

microorganisms (including indicators of faecal contamination e.g. Escherichia coli) suggest hygienic

failure during manufacturing process.
vi © ISO 2017 – All rights reserved
---------------------- Page: 12 ----------------------
SIST EN ISO 18415:2017
INTERNATIONAL STANDARD ISO 18415:2017(E)
Cosmetics — Microbiology — Detection of specified and
non-specified microorganisms
1 Scope

This document gives general guidelines for the detection and identification of specified microorganisms

in cosmetic products as well as for the detection and identification of other kinds of aerobic mesophilic

non-specified microorganisms in cosmetic products.

Microorganisms considered as specified in this document might differ from country to country

according to national practices or regulations. Most of them considered as specified microorganisms

include one or more of the following species: Pseudomonas aeruginosa, Escherichia coli, Staphylococcus

aureus and Candida albicans.

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, hydro-alcoholic products, extreme pH values, etc.

The method described in this document is based on the detection of microbial growth in a non-selective

liquid medium (enrichment broth) suitable to detect microbial contamination, followed by isolation of

microorganisms on non-selective agar media. Other methods can be appropriate depending on the level

of detection required.

In this document specific indications are given for identification of Pseudomonas aeruginosa, Escherichia

coli, Staphylococcus aureus and Candida albicans. Other microorganisms that grow under the conditions

described in this document may be identified by using suitable tests according to a general scheme (see

Annex A). Other standards (e.g. ISO 18416, ISO 21150, ISO 22717, ISO 22718) may be appropriate.

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

be suited in every detail to some products (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.
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 21148:2017, 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:

— IEC Electropedia: available at http:// www .electropedia .org/
— ISO Online browsing platform: available at http:// www .iso .org/ obp
© ISO 2017 – All rights reserved 1
---------------------- Page: 13 ----------------------
SIST EN ISO 18415:2017
ISO 18415:2017(E)
3.1
product
portion of an identified cosmetic product received in the laboratory for testing
3.2
sample

portion of the product (3.1) (at least 1 g or 1 ml) that is used in the test to prepare the initial

suspension (3.3)
3.3
initial suspension

suspension (or solution) of the sample (3.2) in a defined volume of an appropriate enrichment broth (3.8)

3.4
sample dilution
dilution of the initial suspension (3.3)
3.5
aerobic mesophilic microorganism

mesophilic bacterium or yeast growing aerobically under the conditions specified in this document

Note 1 to entry: In the described conditions, other types of microorganism (e.g. moulds) are detectable.

3.6
specified microorganism

aerobic mesophilic bacterium or yeast undesirable in a cosmetic product and recognized as a skin

pathogen species that may be harmful for human health or as an indication of hygienic failure in the

manufacturing process
3.6.1
Pseudomonas aeruginosa
Gram-negative rod (bacilli), motile, smooth colonies pigmented brown or greenish

Note 1 to entry: The main characteristics for identification are growth on a selective cetrimide agar medium,

oxidase positive, production of diffusible fluorescent pigments and production of a soluble phenazine pigment

(pyocyanin) in suitable media.

Note 2 to entry: Pseudomonas aeruginosa can be isolated from a wide variety of environmental sources, especially

in water and has a very high potential to spoil many different substrates. It can produce infections of human skin

or eye areas. It is undesirable in cosmetic products for its potential pathogenicity and its capacity to affect the

physico-chemical properties of the cosmetic formula.
3.6.2
Escherichia coli
Gram-negative rod (bacilli), motile, smooth colonies

Note 1 to entry: The main characteristics are catalase positive, oxidase negative, fermentation of lactose,

production of indole, growth on selective medium containing bile salts with characteristic colonies.

Note 2 to entry: Escherichia coli can be isolated from the moist environmental sources (air, water, soil) and is a

faecal contamination indicator.
3.6.3
Staphylococus aureus

Gram-positive cocci, mainly aggregated in grape-like clusters, smooth colonies generally pigmented

in yellow

Note 1 to entry: The main characteristics for identification are growth on a specific selective medium, catalase

positive, coagulase positive.

Note 2 to entry: Staphylococcus aureus is an opportunistic pathogen for humans, which often can be also present

on the skin of healthy individuals without causing them any apparent illness. It is a specified microorganism and

undesirable in cosmetic products.
2 © ISO 2017 – All rights reserved
---------------------- Page: 14 ----------------------
SIST EN ISO 18415:2017
ISO 18415:2017(E)
3.6.4
Candida albicans

yeast that forms white to beige, creamy and convex colonies on the surface of a non-selective agar medium

Note 1 to entry: The main characteristics for identification are production of germ tube and/or pseudomycelium

and chlamydospore when the test is performed following the method specified in this document.

3.7
non-specified microorganism

aerobic mesophilic bacterium or yeast found in cosmetic products, not defined in 3.6

3.8
enrichment broth

non-selective liquid medium containing suitable neutralizers and/or dispersing agents and

demonstrated to be suitable for the product (3.1) under test
4 Principle

The first step of the procedure is to perform an enrichment by using a non-selective broth medium to

increase the number of microorganisms without the risk of inhibition by the selective ingredients that

are present in selective/differential growth media.

The following steps (isolation and identification) are performed according to need by using appropriate

conditions of incubation and suitable identification test, as described in this document.

The possible inhibition of microbial growth by the sample shall be neutralized to allow the detection

[9]

of viable microorganisms . In all cases and whatever the methodology, the neutralization of the

[9][10][11]
antimicrobial properties of the product shall be checked and demonstrated .
5 Diluents 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 enrichment broth is used to disperse the sample and to increase the initial micro

...

Questions, Comments and Discussion

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