SIST EN ISO 18416:2009
(Main)Cosmetics - Microbiology - Detection of Candida albicans (ISO 18416:2007)
Cosmetics - Microbiology - Detection of Candida albicans (ISO 18416:2007)
This International Standard gives general guidelines for the detection and identification of the specified microorganism Candida albicans in cosmetic products. Microorganisms considered as specified in this International Standard might differ from country to country according to national practices or regulations. In order to ensure product quality and safety for consumers, it is advisable to perform an appropriate microbiological risk analysis so as to determine the types of cosmetic product to which this International Standard is applicable. Products considered to present a low microbiological risk include those with low water activity, hydro-alcoholic products, those with extreme pH values, etc. The method described in this International Standard is based on the detection of Candida albicans in a nonselective liquid medium (enrichment broth), followed by isolation on a selective agar medium. Other methods may be appropriate dependent on the level of detection required.
Kosmetik - Mikrobiologie - Nachweis von Candida albicans (ISO 18416:2007)
Diese Internationale Norm gibt allgemeine Anleitungen für den Nachweis und die Identifizierung des beschrie¬benen Mikroorganismus Candida albicans in kosmetischen Mitteln. Die in dieser Internationalen Norm vorge¬gebenen Mikroorganismen können in Übereinstimmung mit nationalen Praktiken oder Regelungen von Land zu Land unterschiedlich sein.
Um die Qualität des Produkts und die Sicherheit für Verbraucher sicherzustellen, ist es ratsam, eine geeignete mikrobiologische Risikoanalyse zur Bestimmung der Arten von kosmetischen Mitteln durchzuführen, für die diese Internationale Norm anwendbar ist. Produkte, die ein geringes mikrobiologisches Risiko darstellen, umfassen Produkte mit geringer Wasseraktivität, alkoholisch wässrige Produkte, Produkte mit extremen pH Werten usw.
Das in dieser Internationalen Norm beschriebene Verfahren basiert auf dem Nachweis von Candida albicans in einem nicht selektiven Flüssigmedium (Anreicherungsbouillon), gefolgt von einer Isolation auf einem selek¬tiven Agarmedium. In Abhängigkeit von der erforderlichen Nachweisgrenze können andere Verfahren ange¬messen sein.
ANMERKUNG Für den Nachweis von Candida albicans kann eine Subkultivierung auf nicht selektiven Nährmedien erfolgen, gefolgt von geeigneten Schritten zur Identifizierung (z. B. unter Anwendung von Identifizierungskits).
Wegen der großen Vielfalt an kosmetischen Mitteln innerhalb dieses Anwendungsbereichs ist dieses Verfahren möglicherweise für einige Produkte nicht in allen Einzelheiten geeignet (z. B. bestimmte mit Wasser nicht mischbare Produkte). Andere Internationale Normen (z. B. ISO 18415) könnten geeignet sein. Ersatzweise können für die hier aufgeführten Untersuchungen andere Verfahren (z. B. automatisierte) zur Anwendung kommen, vorausgesetzt, dass deren Gleichwertigkeit nach¬gewiesen oder das Verfahren anderweitig validiert wurde.
Cosmétiques - Microbiologie - Détection de Candida albicans (ISO 18416:2007)
L'ISO 18416:2007 donne des lignes directrices générales pour la détection et l'identification du microorganisme spécifié Candida albicans dans les produits cosmétiques. Les microorganismes considérés comme spécifiés dans l'ISO 18416:2007 peuvent différer d'un pays à l'autre, suivant les pratiques ou les réglementations nationales.
La méthode décrite dans l'ISO 18416:2007 repose d'abord sur la détection de Candida albicans dans un milieu liquide non sélectif (milieu liquide d'enrichissement), puis sur l'isolement dudit microorganisme sur un milieu gélosé sélectif. D'autres méthodes peuvent être appropriées en fonction du niveau de détection exigé.
En raison de la grande variété de produits cosmétiques relevant de ce domaine d'application, il se peut que cette méthode ne soit pas adaptée en tous points, à certains produits (par exemple aux produits non miscibles à l'eau). Une autre Norme internationale peut être appropriée. Il est possible de remplacer l'essai présenté ici par d'autres méthodes (par exemple des méthodes automatisées) sous réserve que leur équivalence a été démontrée ou que la méthode a été validée par ailleurs.
Kozmetika - Mikrobiologija - Ugotavljanje prisotnosti kvasovke Candida albicans (ISO 18416:2007)
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN ISO 18416:2009
01-oktober-2009
Kozmetika - Mikrobiologija - Ugotavljanje prisotnosti kvasovke Candida albicans
(ISO 18416:2007)
Cosmetics - Microbiology - Detection of Candida albicans (ISO 18416:2007)
Kosmetik - Mikrobiologie - Nachweis von Candida albicans (ISO 18416:2007)
Cosmétiques - Microbiologie - Détection de Candida albicans (ISO 18416:2007)
Ta slovenski standard je istoveten z: EN ISO 18416:2009
ICS:
07.100.99 Drugi standardi v zvezi z Other standards related to
mikrobiologijo microbiology
71.100.70 .R]PHWLND7RDOHWQL Cosmetics. Toiletries
SULSRPRþNL
SIST EN ISO 18416:2009 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN ISO 18416:2009
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SIST EN ISO 18416:2009
EUROPEAN STANDARD
EN ISO 18416
NORME EUROPÉENNE
EUROPÄISCHE NORM
June 2009
ICS 07.100.99; 71.100.70
English Version
Cosmetics - Microbiology - Detection of Candida albicans (ISO
18416:2007)
Cosmétiques - Microbiologie - Détection de Candida Kosmetik - Mikrobiologie - Nachweis von Candida albicans
albicans (ISO 18416:2007) (ISO 18416:2007)
This European Standard was approved by CEN on 23 May 2009.
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 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 Management Centre has the same status as the
official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,
France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2009 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 18416:2009: E
worldwide for CEN national Members.
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SIST EN ISO 18416:2009
EN ISO 18416:2009 (E)
Contents Page
Foreword .3
2
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SIST EN ISO 18416:2009
EN ISO 18416:2009 (E)
Foreword
The text of ISO 18416:2007 has been prepared by Technical Committee ISO/TC 217 “Cosmetics” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 18416:2009 by
Technical Committee CEN/SS H99 “Products for household and leisure use - Undetermined” the secretariat of
which is held by CMC.
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 2009, and conflicting national standards shall be withdrawn
at the latest by December 2009.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
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, Cyprus, Czech
Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia,
Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain,
Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 18416:2007 has been approved by CEN as a EN ISO 18416:2009 without any modification.
3
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SIST EN ISO 18416:2009
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SIST EN ISO 18416:2009
INTERNATIONAL ISO
STANDARD 18416
First edition
2007-07-15
Cosmetics — Microbiology — Detection
of Candida albicans
Cosmétiques — Microbiologie — Détection de Candida albicans
Reference number
ISO 18416:2007(E)
©
ISO 2007
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SIST EN ISO 18416:2009
ISO 18416:2007(E)
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ii © ISO 2007 – All rights reserved
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SIST EN ISO 18416:2009
ISO 18416:2007(E)
Contents Page
Foreword. iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions. 1
4 Principle. 2
5 Diluents and culture media. 2
5.1 General. 2
5.2 Diluent for the yeast suspension (tryptone sodium chloride solution) . 3
5.3 Culture media . 3
6 Apparatus and glassware . 5
7 Strains 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 in the enrichment broth . 6
9.3 Incubation of the inoculated enrichment broth . 7
9.4 Detection and identification of Candida albicans . 7
10 Expression of the results (detection of Candida albicans) . 8
11 Neutralization of the antimicrobial properties of the product. 8
11.1 General. 8
11.2 Preparation of inoculum . 8
11.3 Validation of the detection method. 9
12 Test report . 9
Annex A (informative) Other media . 11
Annex B (informative) Neutralizers of antimicrobial activity of preservatives and rinsing liquids . 15
Bibliography . 16
© ISO 2007 – All rights reserved iii
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SIST EN ISO 18416:2009
ISO 18416:2007(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.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
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.
ISO 18416 was prepared by Technical Committee ISO/TC 217, Cosmetics.
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SIST EN ISO 18416:2009
ISO 18416:2007(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 Staphylococcus aureus, Pseudomonas aeruginosa
and Candida albicans may be relevant because they can cause skin or eye infections. The detection of other
kinds of microorganism might be of interest since those microorganisms (including indicators of faecal
contamination, e.g. Escherichia coli) suggest hygienic failure during the manufacturing process.
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SIST EN ISO 18416:2009
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SIST EN ISO 18416:2009
INTERNATIONAL STANDARD ISO 18416:2007(E)
Cosmetics — Microbiology — Detection of Candida albicans
1 Scope
This International Standard gives general guidelines for the detection and identification of the specified
microorganism Candida albicans in cosmetic products. Microorganisms considered as specified in this
International Standard might differ from country to country according to national practices or regulations.
In order to ensure product quality and safety for consumers, it is advisable to perform an appropriate
microbiological risk analysis so as to determine the types of cosmetic product to which this International
Standard is applicable. Products considered to present a low microbiological risk include those with low water
activity, hydro-alcoholic products, those with extreme pH values, etc.
The method described in this International Standard is based on the detection of Candida albicans in a non-
selective liquid medium (enrichment broth), followed by isolation on a selective agar medium. Other methods
may be appropriate dependent on the level of detection required.
NOTE For the detection of Candida albicans, subcultures can be performed on non-selective culture media followed
by suitable identification steps (e.g. using identification kits).
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 International Standards
(e.g. ISO 18415) might be appropriate. Other methods (e.g. automated) can be substituted for the test
presented here provided that their equivalence has been demonstrated or the method has been otherwise
validated.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
ISO 21148:2005, Cosmetics — Microbiology — General instructions for microbiological examination
EN 12353, Chemical disinfectants and antiseptics — Preservation of test organisms used for the
determination of bactericidal, mycobactericidal, sporicidal and fungicidal activity
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
product
portion of an identified cosmetic product received in the laboratory for testing
3.2
sample
portion of the product (at least 1 g or 1 ml) that is used in the test to prepare the initial suspension
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SIST EN ISO 18416:2009
ISO 18416:2007(E)
3.3
initial suspension
suspension (or solution) of the sample in a defined volume of an appropriate enrichment broth
3.4
sample dilution
dilution of the initial suspension
3.5
specified microorganisms
aerobic mesophilic bacteria or yeast that is undesirable in a cosmetic product because it can cause skin or
eye infections or is an indication of hygienic failure
3.6
Candida albicans
yeast that form white to beige, creamy and convex colonies on the surface of a selective medium
NOTE The main characteristic for identification is the production of germ tube and/or pseudomycelium and
chlamydospore when the test is performed following the method specified in this International Standard.
3.7
enrichment broth
non-selective liquid medium containing suitable neutralizers and/or dispersing agents and validated for the
product 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 second step (isolation) of the test is performed on a selective medium followed by identification tests.
To prevent the possible inhibition of microbial growth by the sample it shall be neutralized to allow the
[1]
detection of viable microorganisms . In all cases and whatever the methodology, the neutralization of the
[2], [3], [4]
antimicrobial properties of the product shall be checked and validated .
5 Diluents and culture media
5.1 General
General instructions are given in ISO 21148. When water is mentioned in this International Standard, 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 microbial population. It may
contain neutralizers if the specimen to be tested has antimicrobial properties. The efficacy of the neutralization
shall be demonstrated (see Clause 11). Information relative to suitable neutralizers is given in Annex B.
The enrichment broth (5.3.3.1), or any of the ones listed in Annex A, is suitable for checking the presence of
Candida albicans in accordance with this International Standard provided that it is validated in accordance
with Clause 11.
Other diluents and culture media may be used if it has been demonstrated that they are suitable for use.
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SIST EN ISO 18416:2009
ISO 18416:2007(E)
5.2 Diluent for the yeast suspension (tryptone sodium chloride solution)
5.2.1 General
The diluent is used for the preparation of yeast suspension used for the validation procedure (see Clause 11).
5.2.2 Composition
⎯ tryptone, pancreatic digest of casein 1,0 g
⎯ sodium chloride 8,5 g
⎯ water 1 000 ml
5.2.3 Preparation
Dissolve the components in water by mixing whilst heating. Dispense into suitable containers. Sterilize in the
autoclave at 121 °C for 15 min.
After sterilization and cooling of the solution, the pH shall be equivalent to 7,0 ± 0,2 when measured at room
temperature.
5.3 Culture media
5.3.1 General
Culture media may be prepared using the descriptions provided below or from dehydrated culture media in
accordance with the manufacturer's instructions. The instructions provided by the supplier of the media should
be followed.
NOTE Ready-to-use media may be used when their composition and/or growth yields are comparable to those of the
formulae given herein.
5.3.2 Agar medium for validation
5.3.2.1 Sabouraud dextrose agar (SDA)
5.3.2.1.1 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
5.3.2.1.2 Preparation
Dissolve the components or the dehydrated complete medium in the water by 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.
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SIST EN ISO 18416:2009
ISO 18416:2007(E)
5.3.2.2 Other agar media for validation
Other agar media for validation may be used as appropriate (see Annex A).
5.3.3 Enrichment broth
5.3.3.1 Eugon LT 100 broth
5.3.3.1.1 General
This medium contains ingredients which neutralize inhibitory substances present in the sample: lecithin and
polysorbate 80, and dispersing agent octoxynol 9.
5.3.3.1.2 Composition
⎯ pancreatic digest of casein 15,0 g
⎯ papaic digest of soybean meal 5,0 g
⎯ L-cystine 0,7 g
⎯ sodium chloride 4,0 g
⎯ sodium sulfite 0,2 g
⎯ glucose 5,5 g
⎯ egg lecithin 1,0 g
⎯ polysorbate 80 5,0 g
⎯ octoxynol 9 1,0 g
⎯ water 1 000 ml
5.3.3.1.3 Preparation
Dissolve the components polysorbate 80, octoxynol 9 and egg lecithin one after another in boiling water to
complete dissolution. Dissolve the other components by mixing whilst heating. Dispense the medium into
suitable containers. Sterilize in the autoclave at 121 °C for 15 min.
After sterilization and cooling of the solution, the pH shall be equivalent to 7,0 ± 0,2 when measured at room
temperature.
5.3.3.2 Other enrichment broths
Other enrichment broths may be used as appropriate (see Annex A).
5.3.4 Selective agar medium for isolation of Candida albicans
5.3.4.1 Sabouraud dextrose chloramphenicol agar
5.3.4.1.1 Composition
⎯ dextrose 40,0 g
⎯ peptic digest of animal tissue 5,0 g
⎯ pancreatic digest of casein 5,0 g
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SIST EN ISO 18416:2009
ISO 18416:2007(E)
⎯ chloramphenicol 0,050 g
⎯ agar 15,0 g
⎯ water 1 000 ml
5.3.4.1.2 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 at 121 °C for
15 min. After sterilization the pH shall be equivalent to 5,6 ± 0,2 when measured at room temperature.
5.3.4.2 Other selective agar media
Other selective agar media may be used as appropriate (see Annex A).
5.3.5 Corn meal agar with 1 % polysorbate 80
5.3.5.1 Com
...
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