ISO 15819:2008
(Main)Cosmetics — Analytical methods — Nitrosamines: Detection and determination of N-nitrosodiethanolamine (NDELA) in cosmetics by HPLC-MS-MS
Cosmetics — Analytical methods — Nitrosamines: Detection and determination of N-nitrosodiethanolamine (NDELA) in cosmetics by HPLC-MS-MS
ISO 15819:2008 describes a method for the detection and quantification of NDELA in cosmetics and raw materials used in cosmetics. This method is not applicable to the detection and/or quantification of nitrosamines other than NDELA nor to the detection and/or quantification of NDELA in products other than cosmetics or raw materials used in cosmetics.
Cosmétiques — Méthodes analytiques — Nitrosamines: Recherche et dosage de la N-nitrosodiéthanolamine (NDELA) dans les produits cosmétiques par CLHP-SM-SM
L'ISO 15819:2008 décrit une méthode de recherche et de quantification de la NDELA dans les cosmétiques et les matières premières utilisées dans les cosmétiques. La présente méthode n'est applicable ni à la recherche et/ou à la quantification de nitrosamines autres que la NDELA, ni à la recherche et/ou à la quantification de la NDELA dans les produits autres que les cosmétiques ou les matières premières utilisées dans les cosmétiques.
Kozmetika - Analizne metode - Nitrozamini: detekcija in določevanje N-nitrozodietanolamina (NDELA) v kozmetičnih izdelkih s tekočinsko kromatografijo visoke ločljivosti z masno spektrometrijsko detekcijo (HPLC-MS-MS)
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МЕЖДУНАРОДНЫЙ ISO
СТАНДАРТ 15819
Первое издание
2008-06-01
Косметика. Аналитические методы.
Нитрозамины: Обнаружение и
определение содержания N-
нитрозодиэтаноламина (NDELA) в
косметике методом жидкостной
хроматографии высокого
разрешенения одновременно с масс-
спектрометрическим обнаружением
(HPLC-MS-MS)
Cosmetics – Analytical methods – Nitrosamines: Detection and
determination of N-nitrosodiethanolamine (NDELA in cosmetics)by
HPLC-MS-MS
Ответственность за подготовку русской версии несёт GOST R
(Российская Федерация) в соответствии со статьёй 18.1 Устава
Ссылочный номер
ISO 15819:2008(R)
©
ISO 2008
---------------------- Page: 1 ----------------------
ISO 15819:2008(R)
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ISO 15819:2008(R)
Предисловие
Международная организация по стандартизации (ISO) является всемирной федерацией национальных
организаций по стандартизации (комитетов-членов ISO). Разработка международных стандартов
обычно осуществляется техническими комитетами ISO. Каждый комитет-член, заинтересованный в
деятельности, для которой был создан технический комитет, имеет право быть представленным в этом
комитете. Международные правительственные и неправительственные организации, имеющие связи с
ISO, также принимают участие в работах. ISO работает в тесном сотрудничестве с Международной
электротехнической комиссией (IEC) по всем вопросам стандартизации в области электротехники.
Международные стандарты разрабатываются в соответствии с правилами, установленными в
Директивах ISO/IEC, Часть 2.
Основная задача технических комитетов состоит в подготовке международных стандартов. Проекты
международных стандартов, одобренные техническими комитетами, рассылаются комитетам-членам
на голосование. Их опубликование в качестве международных стандартов требует одобрения, по
меньшей мере, 75 % комитетов-членов, принимающих участие в голосовании.
Следует иметь в виду, что некоторые элементы этого документа могут быть объектом патентных прав.
Организация ISO не должна нести ответственность за идентификацию какого-либо одного или всех
патентных прав.
ISO 15819 подготовлен Техническим Комитетом ISO/TC 217, Косметические средства.
© ISO 2008 – Все права сохраняются iii
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ISO 15819:2008(R)
Введение
Воздействие на человека N-нитрозаминов может происходить из разных источников таких, как
окружающая среда, продукты питания или продукты личной гигиены. В результате действия их
канцерогенного потенциала на некоторые животные виды, для сохранения здоровья человека
считается очень важным свести к минимуму воздействие N-нитрозаминов. Среди N-нитрозаминов, N-
нитрозодиэтаноламин (NDELA) признан потенциальным загрязнителем косметики.
В данном контексте разработаны несколько аналитических методов для обнаружения и определения
его в косметике. Разработан анализ методом газовой хроматографии/тепловой энергии, методом
жидкостной хроматографии высокого разрешения (HPLC) в сочетании либо с фотолизом и
колориметрической оценкой, либо с определением масс-спектрометрией (MS). Этот последний метод
использует передовую технологию для обеспечения максимальной специфичности в отношении
NDELA, минимизирует риск артифактного образования интересующего аналита и позволяет провести
точную оценку количества.
Данный аналитический метод использует жидкостную хроматографию высокого разрешения совместно
с масс-спектрометрией для отделения и обнаружения на уровне следов NDELA из косметического
ингредиента или матрицы продукта с максимальной специфичностью для NDELA.
iv © ISO 2008 – Все права сохраняются
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МЕЖДУНАРОДНЫЙ СТАНДАРТ ISO 15819:2008(R)
Косметика. Аналитические методы. Нитрозамины:
Обнаружение и определение содержания N-
нитрозодиэтаноламина (NDELA) в косметике методом
жидкостной хроматографии высокого разрешенения
одновременно с масс-спектрометрическим обнаружением
(HPLC-MS-MS)
1 Область применения
Настоящий международный стандарт описывает метод обнаружения и количественного определения
N-нитрозодиэтаноламина (NDELA) в косметических средствах и сырьевых материалах, применяемых в
косметике.
Данный метод не применяется ни для обнаружения и/или количественного определения нитрозаминов
в иных, чем NDELA, ни для обнаружения и/или количественного определения NDELA в продуктах,
отличных от косметических средств или сырьевых материалов, применяемых в косметике.
Если продукт имеет возможность загрязнения либо NDELA ингредиентами, либо в результате
образования NDELA составом ингредиентов, этот метод должен применяться для количественного
определения NDELA. Соответственно, этот метод не следует применять для обычных типовых
испытаний косметических изделий. По причине широкого разнообразия косметической продукции в
данной области применения, для этого метода может потребоваться адаптация к некоторым матрицам.
Поэтому отдельно разрабатывается международный стандарт, предназначенный для альтернативных
методов тестирования нитрозаминов в косметических продуктах. Могут применяться и другие методы
при условии, что они проверены на способность обнаружения NDELA и подтверждены в вопросах
восстановления и количественного определения аналита.
2 Нормативные ссылки
Следующие ссылочные нормативные документы являются обязательными для применения настоящего документа.
Для жестких ссылок применяется только цитируемое издание документа. Для плавающих ссылок необходимо
использовать самое последнее издание нормативного ссылочного документа (включая любые изменения).
ISO 3696:1987, Вода для лабораторного анализа. Технические требования и методы испытания
3 Принцип
Экстракция нитрозамина NDELA в косметических пробах выполняется водой в присутствии
дейтерированного d8-NDELA, используемого в качестве внутреннего стандарта. Очистка выполняется
либо с помощью экстракции в твердой фазе (очистка SPE, см. 6.3.1) с картриджем C18 или жидкость-
жидкостной экстракцией с помощью дихлорметана (очистка CM, см.3.2), когда пробы не растворимы в
воде. Экстракты анализируются методом HPLC-MS-MS (жидкостной хроматографии высокого
разрешения одновременно с масс-спектрометрическим обнаружением).
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ISO 15819:2008(R)
Количественный анализ NDELA производится путем сравнения отношения ионов главных фрагментов
NDELA и d8-NDELA с калибровочной кривой.
Подтверждение присутствия NDELA выполняется с помощью молекулярного иона или двух
диагностических ионов.
4 Реактивы
Во время проведения анализа, если не заявлено иначе, используют только реактивы признанного
аналитического качества и только дистиллированную воду или воду класса 1 в соответствии с
ISO 3696:1987. Растворитель должен быть качества соответствующего анализу HPLC (жидкостной
хроматографии высокого разрешения).
4.1 Метанол (MeOH), качество для HPLC.
4.2 Этанол (EtOH), качество для HPLC.
4.3 Дихлорметан, качество для HPLC.
4.4 N-нитрозодиэтаноламин, с известной чистотой более чем 95 %.
4.5 d8-N- нитрозодиэтаноламин, с известной чистотой более чем 95 %.
4.6 Ацетат аммония (NH Ac), аналитического качества.
4
4.7 Раствор ацетата аммония 1 моль/л, получен растворением 77,08 г NH Ac в 1,0 л воды.
4
4.8 Элюент A: 2 ммоль NH Ac в воде, получен отбором 2 мл из раствора 1 моль/л NH Ac (4.7) и
4 4
разведенный до 1 л водой.
4.9 Элюент B: 2 ммоль NH Ac в 90 % MeOH/вода, получен отбором 2 мл из раствора 1 моль/л
4
NH Ac (4.7) и дополнен 900 мл MeOH и 98 мл воды.
4
5 Аппаратура
Используется стандартная лабораторная посуда и оборудование с добавлением:
5.1 Миксер Vortex.
5.2 Станция обработки проб, в случае применения SPE (такая, станция обработки проб как
®1)
Vacmaster , IST).
5.3 Центрифуга, способная достигать не менее 20 000 G.
1)
®
5.4 Колонки экстракции твердой фазы, т. н. Bakerbond C18 – 6 мл, 500 мг обращенной фазы
октадецилсилана, связанного с силикагелем, 40 APD, 60 Å.
5.5 Оборудование HPLC-MS-MS, состоит из:
® ® ®
1) Vacmaster , Bakerbond и Spherisorb являются примерами подходящих изделий, имеющихся в продаже.
Данная информация представлена для удобства пользователей настоящего международного стандарта и не
является подтверждением ISO данной продукции.
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ISO 15819:2008(R)
5.5.1 Аппаратура жидкостной хроматографии высокого разрешения, состоящая из резервуара с
элюентом, насоса, системы впрыскивания, процессора обработки данных, т. н. интегратора с
плоттером, соединенная с тандемным масс-спектрометром, использующим ионизацию
...
INTERNATIONAL ISO
STANDARD 15819
First edition
2008-06-01
Cosmetics — Analytical methods —
Nitrosamines: Detection and
determination of N-nitrosodiethanolamine
(NDELA) in cosmetics by HPLC-MS-MS
Cosmétiques — Méthodes analytiques — Nitrosamines: Recherche et
dosage des N-nitrosodiéthanolamines (NDELA) dans les produits
cosmétiques par CLHP-SM-SM
Reference number
ISO 15819:2008(E)
©
ISO 2008
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ISO 15819:2008(E)
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Published in Switzerland
ii © ISO 2008 – All rights reserved
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ISO 15819:2008(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 15819 was prepared by Technical Committee ISO/TC 217, Cosmetics.
© ISO 2008 – All rights reserved iii
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ISO 15819:2008(E)
Introduction
Human exposure to N-nitrosamines can occur through diverse sources such as environment, food or personal
care products. As a result of their perceived carcinogenic potential on several animal species, minimization of
exposure to N-nitrosamines is recognised as important to the preservation of human health. Among
N-nitrosamines, N-nitrosodiethanolamine (NDELA) has been recognised as a potential contaminant of
cosmetics.
In this context, several analytical methods have been developed to detect and determine its presence in
cosmetics – such as gas chromatography/thermal energy analysis, high performance liquid chromatography
(HPLC) coupled either with photolysis and colorimetric quantification or with mass spectrometry (MS)
determination. This latter method uses advanced technology to ensure the maximum specificity towards
NDELA, to minimize the risk of artifactual formation of the analyte of interest and to allow precise
quantification.
This analytical method uses high performance liquid chromatography coupled with mass spectrometry to
separate and detect trace levels of NDELA from a cosmetic ingredient or product matrix with maximum
specificity for NDELA.
iv © ISO 2008 – All rights reserved
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INTERNATIONAL STANDARD ISO 15819:2008(E)
Cosmetics — Analytical methods — Nitrosamines: Detection
and determination of N-nitrosodiethanolamine (NDELA) in
cosmetics by HPLC-MS-MS
1 Scope
This International Standard describes a method for the detection and quantification of NDELA in cosmetics
and raw materials used in cosmetics.
This method is not applicable to the detection and/or quantification of nitrosamines other than NDELA nor to
the detection and/or quantification of NDELA in products other than cosmetics or raw materials used in
cosmetics.
If a product has a possibility of either NDELA contamination from ingredients or NDELA formation by the
composition of ingredients, the method shall be applied for quantitative determination of NDELA. Accordingly
the method would not be applied in routine testing of cosmetic products. Because of the large variety of
cosmetic products within this field of application, this method might need to be adapted for certain matrices.
Therefore, International Standards dedicated to alternative methods for testing nitrosamines in cosmetic
products are being developed separately. Other methods can be employed provided that they are verified as
to their detection of NDELA and validated in terms of recovery and quantification of the analyte.
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 3696:1987, Water for analytical laboratory use — Specification and test methods
3 Principle
Extraction of the nitrosamine NDELA in cosmetic samples is carried out with water in the presence of
deuterated d8-NDELA used as internal standard. Clean-up is performed either using solid phase extraction
(SPE clean-up, see 6.3.1) with a C18 cartridge or liquid–liquid extraction using dichloromethane (DCM
clean-up, see 6.3.2) when the samples are not dispersible in water. The extracts are analysed by HPLC-MS-
MS (high performance liquid chromatography coupled with tandem mass spectrometric detection).
NDELA quantification is done by comparing the ratio of the major fragment ions of NDELA and d8-NDELA
with the calibration curve.
Confirmation of the presence of NDELA is carried out by using the molecular ion and two diagnostic ions.
© ISO 2008 – All rights reserved 1
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ISO 15819:2008(E)
4 Reagents
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and only
distilled water or water of grade 1 in accordance with ISO 3696:1987. Solvent shall be of quality for HPLC
analysis.
4.1 Methanol (MeOH), HPLC grade.
4.2 Ethanol (EtOH), HPLC grade.
4.3 Dichloromethane, HPLC grade.
4.4 N-nitrosodiethanolamine, with known purity greater than 95 %.
4.5 d8-N-nitrosodiethanolamine, with known purity greater than 95 %.
4.6 Ammonium acetate (NH Ac), analytical grade.
4
4.7 1 mol/l ammonium acetate solution, formed by dissolving 77,08 g of NH Ac in 1,0 l water.
4
4.8 Eluent A: 2 mmol NH Ac in water, formed by taking 2 ml of 1 mol/l NH Ac (4.7) and making up to 1 l
4 4
with water.
4.9 Eluent B: 2 mmol NH Ac in 90 % MeOH/water, formed by taking 2 ml of 1 mol/l NH Ac (4.7) and
4 4
adding 900 ml MeOH and 98 ml water.
5 Apparatus
Use standard laboratory glassware and equipment, with the addition of:
5.1 Vortex mixer.
®1)
5.2 Sample processing station, in SPE application (such as Vacmaster sample processing station,
IST).
5.3 Centrifuge, capable of reaching not less than 20 000 G.
1)
®
5.4 Solid phase extraction columns, e.g. Bakerbond C18 – 6 ml, 500 mg reversed phase
octadecylsilane bonded to silicagel, 40 APD, 60 Å.
5.5 HPLC-MS-MS equipment consisting of:
5.5.1 High performance liquid chromatography apparatus, consisting of an eluent reservoir, a pump, an
injection system, a data processor, e.g. an integrator with plotter, coupled with tandem mass spectrometry
using electrospray ionization.
® ® ®
1) Vacmaster , Bakerbond and Spherisorb are examples of suitable products available commercially. This
information is given for the convenience of users of this International Standard and does not constitute an endorsement by
ISO of these products.
2 © ISO 2008 – All rights reserved
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ISO 15819:2008(E)
1)
®
5.5.2 Analytical reversed phase HPLC separating column, C18, e.g. Spherisorb ODS II protected
with a guard column, the dimensions of which are:
separating column
⎯ length: 150 mm
⎯ internal diameter: 4,6 mm
⎯ size of spherical particles: 5 µm
guard column
⎯ length: 10 mm
⎯ internal di
...
SLOVENSKI STANDARD
SIST ISO 15819:2009
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YLVRNHORþOMLYRVWL]PDVQRVSHNWURPHWULMVNRGHWHNFLMR +3/&0606
Cosmetics - Analytical methods - Nitrosamines: Detection and determination of N-
nitrosodiethanolamine (NDELA) in cosmetics by HPLC-MS-MS
Cosmétiques - Méthodes analytiques - Nitrosamines: Recherche et dosage des N-
nitrosodiéthanolamines (NDELA) dans les produits cosmétiques par CLHP-SM-SM
Ta slovenski standard je istoveten z: ISO 15819:2008
ICS:
71.100.70 .R]PHWLND7RDOHWQL Cosmetics. Toiletries
SULSRPRþNL
SIST ISO 15819:2009 en,fr
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
---------------------- Page: 1 ----------------------
SIST ISO 15819:2009
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SIST ISO 15819:2009
INTERNATIONAL ISO
STANDARD 15819
First edition
2008-06-01
Cosmetics — Analytical methods —
Nitrosamines: Detection and
determination of N-nitrosodiethanolamine
(NDELA) in cosmetics by HPLC-MS-MS
Cosmétiques — Méthodes analytiques — Nitrosamines: Recherche et
dosage des N-nitrosodiéthanolamines (NDELA) dans les produits
cosmétiques par CLHP-SM-SM
Reference number
ISO 15819:2008(E)
©
ISO 2008
---------------------- Page: 3 ----------------------
SIST ISO 15819:2009
ISO 15819:2008(E)
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COPYRIGHT PROTECTED DOCUMENT
© ISO 2008
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or
ISO's member body in the country of the requester.
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Published in Switzerland
ii © ISO 2008 – All rights reserved
---------------------- Page: 4 ----------------------
SIST ISO 15819:2009
ISO 15819:2008(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 15819 was prepared by Technical Committee ISO/TC 217, Cosmetics.
© ISO 2008 – All rights reserved iii
---------------------- Page: 5 ----------------------
SIST ISO 15819:2009
ISO 15819:2008(E)
Introduction
Human exposure to N-nitrosamines can occur through diverse sources such as environment, food or personal
care products. As a result of their perceived carcinogenic potential on several animal species, minimization of
exposure to N-nitrosamines is recognised as important to the preservation of human health. Among
N-nitrosamines, N-nitrosodiethanolamine (NDELA) has been recognised as a potential contaminant of
cosmetics.
In this context, several analytical methods have been developed to detect and determine its presence in
cosmetics – such as gas chromatography/thermal energy analysis, high performance liquid chromatography
(HPLC) coupled either with photolysis and colorimetric quantification or with mass spectrometry (MS)
determination. This latter method uses advanced technology to ensure the maximum specificity towards
NDELA, to minimize the risk of artifactual formation of the analyte of interest and to allow precise
quantification.
This analytical method uses high performance liquid chromatography coupled with mass spectrometry to
separate and detect trace levels of NDELA from a cosmetic ingredient or product matrix with maximum
specificity for NDELA.
iv © ISO 2008 – All rights reserved
---------------------- Page: 6 ----------------------
SIST ISO 15819:2009
INTERNATIONAL STANDARD ISO 15819:2008(E)
Cosmetics — Analytical methods — Nitrosamines: Detection
and determination of N-nitrosodiethanolamine (NDELA) in
cosmetics by HPLC-MS-MS
1 Scope
This International Standard describes a method for the detection and quantification of NDELA in cosmetics
and raw materials used in cosmetics.
This method is not applicable to the detection and/or quantification of nitrosamines other than NDELA nor to
the detection and/or quantification of NDELA in products other than cosmetics or raw materials used in
cosmetics.
If a product has a possibility of either NDELA contamination from ingredients or NDELA formation by the
composition of ingredients, the method shall be applied for quantitative determination of NDELA. Accordingly
the method would not be applied in routine testing of cosmetic products. Because of the large variety of
cosmetic products within this field of application, this method might need to be adapted for certain matrices.
Therefore, International Standards dedicated to alternative methods for testing nitrosamines in cosmetic
products are being developed separately. Other methods can be employed provided that they are verified as
to their detection of NDELA and validated in terms of recovery and quantification of the analyte.
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 3696:1987, Water for analytical laboratory use — Specification and test methods
3 Principle
Extraction of the nitrosamine NDELA in cosmetic samples is carried out with water in the presence of
deuterated d8-NDELA used as internal standard. Clean-up is performed either using solid phase extraction
(SPE clean-up, see 6.3.1) with a C18 cartridge or liquid–liquid extraction using dichloromethane (DCM
clean-up, see 6.3.2) when the samples are not dispersible in water. The extracts are analysed by HPLC-MS-
MS (high performance liquid chromatography coupled with tandem mass spectrometric detection).
NDELA quantification is done by comparing the ratio of the major fragment ions of NDELA and d8-NDELA
with the calibration curve.
Confirmation of the presence of NDELA is carried out by using the molecular ion and two diagnostic ions.
© ISO 2008 – All rights reserved 1
---------------------- Page: 7 ----------------------
SIST ISO 15819:2009
ISO 15819:2008(E)
4 Reagents
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and only
distilled water or water of grade 1 in accordance with ISO 3696:1987. Solvent shall be of quality for HPLC
analysis.
4.1 Methanol (MeOH), HPLC grade.
4.2 Ethanol (EtOH), HPLC grade.
4.3 Dichloromethane, HPLC grade.
4.4 N-nitrosodiethanolamine, with known purity greater than 95 %.
4.5 d8-N-nitrosodiethanolamine, with known purity greater than 95 %.
4.6 Ammonium acetate (NH Ac), analytical grade.
4
4.7 1 mol/l ammonium acetate solution, formed by dissolving 77,08 g of NH Ac in 1,0 l water.
4
4.8 Eluent A: 2 mmol NH Ac in water, formed by taking 2 ml of 1 mol/l NH Ac (4.7) and making up to 1 l
4 4
with water.
4.9 Eluent B: 2 mmol NH Ac in 90 % MeOH/water, formed by taking 2 ml of 1 mol/l NH Ac (4.7) and
4 4
adding 900 ml MeOH and 98 ml water.
5 Apparatus
Use standard laboratory glassware and equipment, with the addition of:
5.1 Vortex mixer.
®1)
5.2 Sample processing station, in SPE application (such as Vacmaster sample processing station,
IST).
5.3 Centrifuge, capable of reaching not less than 20 000 G.
1)
®
5.4 Solid phase extraction columns, e.g. Bakerbond C18 – 6 ml, 500 mg reversed phase
octadecylsilane bonded to silicagel, 40 APD, 60 Å.
5.5 HPLC-MS-MS equipment consisting of:
5.5.1 High performance liquid chromatography apparatus, consisting of an eluent reservoir, a pump, an
injection system, a data processor, e.g. an integrator with plotter, coupled with tandem mass spectrometry
using electrospray ionization.
® ® ®
1) Vacmaster , Bakerbond and Spherisorb are examples of suitable products available commercially. This
information is give
...
NORME ISO
INTERNATIONALE 15819
Première édition
2008-06-01
Version corrigée
2008-08-15
Cosmétiques — Méthodes analytiques —
Nitrosamines: Recherche et dosage de la
N-nitrosodiéthanolamine (NDELA) dans
les produits cosmétiques par
CLHP-SM-SM
Cosmetics — Analytical methods — Nitrosamines: Detection and
determination of N-nitrosodiethanolamine (NDELA) in cosmetics by
HPLC-MS-MS
Numéro de référence
ISO 15819:2008(F)
©
ISO 2008
---------------------- Page: 1 ----------------------
ISO 15819:2008(F)
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Publié en Suisse
ii © ISO 2008 – Tous droits réservés
---------------------- Page: 2 ----------------------
ISO 15819:2008(F)
Avant-propos
L'ISO (Organisation internationale de normalisation) est une fédération mondiale d'organismes nationaux de
normalisation (comités membres de l'ISO). L'élaboration des Normes internationales est en général confiée
aux comités techniques de l'ISO. Chaque comité membre intéressé par une étude a le droit de faire partie du
comité technique créé à cet effet. Les organisations internationales, gouvernementales et non
gouvernementales, en liaison avec l'ISO participent également aux travaux. L'ISO collabore étroitement avec
la Commission électrotechnique internationale (CEI) en ce qui concerne la normalisation électrotechnique.
Les Normes internationales sont rédigées conformément aux règles données dans les Directives ISO/CEI,
Partie 2.
La tâche principale des comités techniques est d'élaborer les Normes internationales. Les projets de Normes
internationales adoptés par les comités techniques sont soumis aux comités membres pour vote. Leur
publication comme Normes internationales requiert l'approbation de 75 % au moins des comités membres
votants.
L'attention est appelée sur le fait que certains des éléments du présent document peuvent faire l'objet de
droits de propriété intellectuelle ou de droits analogues. L'ISO ne saurait être tenue pour responsable de ne
pas avoir identifié de tels droits de propriété et averti de leur existence.
L'ISO 15819 a été élaborée par le comité technique ISO/TC 217, Cosmétiques.
La présente version corrigée de l'ISO 15819:2008, inclut une modification du titre ainsi que quelques
modifications rédactionnelles.
© ISO 2008 – Tous droits réservés iii
---------------------- Page: 3 ----------------------
ISO 15819:2008(F)
Introduction
L'exposition humaine aux N-nitrosamines peut se produire au contact de sources diverses présentes dans
l'environnement, la nourriture ou les produits de soin personnels. Ces substances ayant montré un effet
cancérigène notoire sur plusieurs espèces animales, il est reconnu qu'une restriction d'exposition aux
N-nitrosamines est d'une importance primordiale pour la préservation de la santé humaine. Parmi les
N-nitrosamines, il a été établi que la N-nitrosodiéthanolamine (NDELA) est un contaminant potentiel des
cosmétiques.
Dans ce contexte, plusieurs méthodes analytiques ont été développées pour rechercher et doser sa présence
dans les cosmétiques, comme la chromatographie en phase gazeuse couplée à l'analyse d'énergie thermique,
la chromatographie en phase liquide à haute performance (CLHP) couplée soit à une photolyse avec
quantification colorimétrique soit à une détermination par spectrométrie de masse (SM). Cette dernière
méthode nécessite l'usage de techniques avancées pour garantir une spécificité maximale dans la recherche
de la NDELA, afin de limiter le risque de formation artefactuelle de l'analyte considéré et de permettre une
quantification précise de la substance.
La présente méthode analytique emploie la chromatographie en phase liquide à haute performance couplée à
la spectrométrie de masse afin de séparer et de rechercher la NDELA à l'état de trace dans un ingrédient ou
dans une matrice de produit cosmétique, avec une spécificité maximale concernant la NDELA.
iv © ISO 2008 – Tous droits réservés
---------------------- Page: 4 ----------------------
NORME INTERNATIONALE ISO 15819:2008(F)
Cosmétiques — Méthodes analytiques — Nitrosamines:
Recherche et dosage de la N-nitrosodiéthanolamine (NDELA)
dans les produits cosmétiques par CLHP-SM-SM
1 Domaine d'application
La présente Norme internationale décrit une méthode de recherche et de quantification de la NDELA dans les
cosmétiques et les matières premières utilisées dans les cosmétiques.
La présente méthode n'est applicable ni à la recherche et/ou à la quantification de nitrosamines autres que la
NDELA, ni à la recherche et/ou à la quantification de la NDELA dans les produits autres que les cosmétiques
ou les matières premières utilisées dans les cosmétiques.
Si un produit peut être contaminé par la NDELA à partir de ses ingrédients ou que la composition
d'ingrédients cosmétiques peut entraîner la formation de NDELA, alors la méthode sera appliquée pour la
détermination de la quantité de NDELA. Par conséquent, cette méthode ne sera pas appliquée dans les
essais de routine relatifs aux produits cosmétiques. En raison du large éventail de cosmétiques considéré
dans ce domaine d'application, la présente méthode pourrait nécessiter d'être adaptée à certaines matrices.
Par voie de conséquence, les Normes internationales consacrées à d'autres méthodes d'essai des
nitrosamines dans les cosmétiques sont élaborées séparément. Ces méthodes peuvent être employées à
condition que la recherche de NDELA ait été vérifiée et que ces méthodes soient validées en termes de
récupération et de quantification de l'analyte.
2 Références normatives
Les documents de référence suivants sont indispensables pour l'application du présent document. Pour les
références datées, seule l'édition citée s'applique. Pour les références non datées, la dernière édition du
document de référence (y compris les éventuels amendements) s'applique.
ISO 3696:1987, Eau pour laboratoire à usage analytique — Spécification et méthodes d'essai
3 Principe
L'extraction de la nitrosamine NDELA dans les échantillons de cosmétiques est réalisée avec de l'eau en
présence de d8-NDELA deutérée utilisée en tant qu'étalon interne. Une purification est effectuée soit par
extraction en phase solide (purification SPE, voir 6.3.1) à l'aide d'une cartouche C18, soit par extraction
liquide-liquide à l'aide de dichlorométhane (purification DCM, voir 6.3.2) lorsque les échantillons ne peuvent
pas se disperser dans l'eau. Les extraits sont analysés par CLHP-SM-SM (chromatographie en phase liquide
à haute performance couplée à une détection spectrométrique de masse).
La quantification de NDELA est réalisée par comparaison de la proportion d'ions fragments majoritaires de
NDELA et de d8-NDELA à l'aide d'une courbe d'étalonnage.
La confirmation de la présence de NDELA est obtenue à l'aide de l'ion moléculaire et de deux ions qualifiants.
© ISO 2008 – Tous droits réservés 1
---------------------- Page: 5 ----------------------
ISO 15819:2008(F)
4 Réactifs
Sauf indication contraire, n'utiliser, au cours de l'analyse, que des réactifs de qualité analytique reconnue et
de l'eau distillée ou de l'eau de qualité 1, conformément à l'ISO 3696:1987. Le solvant doit être de qualité
pour analyse CLHP.
4.1 Méthanol (MeOH), qualité CLHP.
4.2 Éthanol (EtOH), qualité CLHP.
4.3 Dichlorométhane, qualité CLHP.
4.4 N-nitrosodiéthanolamine, de pureté connue, supérieure à 95 %.
4.5 d8-N-nitrosodiéthanolamine, de pureté connue, supérieure à 95 %.
4.6 Acétate d'ammonium (NH Ac), qualité analytique.
4
4.7 Solution d'acétate d'ammonium 1 mol/l, préparée par dissolution de 77,08 g de NH Ac dans 1,0 l
4
d'eau.
4.8 Éluant A: 2 mmol de NH Ac dans l'eau, préparé avec 2 ml de NH Ac à 1 mol/l (4.7), complété à 1,0 l
4 4
avec de l'eau.
4.9 Éluant B: 2 mmol de NH Ac dans 90 % de MeOH/eau, préparé avec 2 ml de NH Ac à 1 mol/l (4.7),
4 4
complété avec 900 ml de MeOH et 98 ml d'eau.
5 Appareillage
Utiliser une verrerie et un équipement de laboratoire normalisés, ainsi que les éléments suivants:
5.1 Agitateur de type vortex.
5.2 Station de traitement d'échantillon, pour application SPE (telle que la station de traitement
1)
d'échantillon Vacmaster® , IST).
5.3 Centrifugeuse, pouvant atteindre au moins 20 000 G.
1)
5.4 Colonnes d'extraction en phase solide, par exemple Bakerbond® C18 – 6 ml, avec 500 mg
d'octadécylsilane en phase inverse fixée sur gel de silice, 40 APD, 60 Å.
5.5 Équipement CLHP-SM-SM composé de
5.5.1 Chromatographe en phase liquide à haute performance, consistant en un réservoir d'éluant, une
pompe, un système d'injection, un traitement des données, par exemple un intégrateur avec traceur, couplé à
un spectromètre de masse à ionisation par électrospray.
1)
Vacmaster®, Bakerbond® et Spherisorb® sont des exemples de produits appropriés disponibles sur le marché. Cette
information est donnée à l'intention des utilisateurs de la présente Norme internationale et ne signifie nullement que l'ISO
approuve ou recommande l'emploi exclusif des produits ainsi désignés.
2 © ISO 2008 – Tous droits réservés
---------------------- Page: 6 ----------------------
ISO 15819:2008(F)
5.5.2 Colonne analytique de séparation CLHP en phase inverse C18, par exemple Spherisorb® ODS
1)
II protégée par une colonne de garde dont les dimensions sont les suivantes:
colonne de séparation
⎯ longueur: 15
...
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Cosmetics - Analytical methods - Nitrosamines: Detection and determination of N-nitrosodiethanolamine (NDELA) in cosmetics by HPLC-MSLMRCosmétiques - Méthodes analytiques - Nitrosamines: Recherche et dosage des N-nitrosodiéthanolamines (NDELA) dans les produits cosmétiques par CLHP-SM71.100.70SULSRPRþNLCosmetics. ToiletriesICS:oSIST ISO/FDIS 15819:2008enTa slovenski standard je istoveten z:01-junij-2008oSIST ISO/FDIS 15819:2008SLOVENSKI
STANDARD
RECIPIENTS OF THIS DRAFT ARE INVITED TOSUBMIT, WITH THEIR COMMENTS, NOTIFICATION OF ANY RELEVANT PATENT RIGHTS OF WHICH THEY ARE AWARE AND TO PROVIDE SUPPORT-ING DOCUMENTATION. IN ADDITION TO THEIR EVALUATION AS BEING ACCEPTABLE FOR INDUSTRIAL, TECHNO-LOGICAL, COMMERCIAL AND USER PURPOSES,DRAFT INTERNATIONAL STANDARDS MAY ONOCCASION HAVE TO BE CONSIDERED IN THELIGHT OF THEIR POTENTIAL TO BECOME STAN-DARDS TO WHICH REFERENCE MAY BE MADE INNATIONAL REGULATIONS.
Reference numberISO/FDIS 15819:2008(E)© ISO 2008 FINAL DRAFT ISO/TC 217 Secretariat: ISIRI Voting begins on: 2008-02-25 Voting terminates on: 2008-04-25
INTERNATIONAL STANDARD ISO/FDIS15819Cosmetics — Analytical methods — Nitrosamines: Detection and determination of N-nitrosodiethanolamine (NDELA) in cosmetics by HPLC-MS Cosmétiques — Méthodes analytiques — Nitrosamines: Recherche et dosage des N-nitrosodiéthanolamines (NDELA) dans les produits cosmétiques par CLHP-SM
ISO/FDIS 15819:2008(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat accepts no liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
Copyright notice This ISO document is a Draft International Standard and is copyright-protected by ISO. Except as permitted under the applicable laws of the user's country, neither this ISO draft nor any extract from it may be reproduced, stored in a retrieval system or transmitted in any form or by any means, electronic, photocopying, recording or otherwise, without prior written permission being secured. Requests for permission to reproduce should be addressed to either ISO at the address below or ISO's member body in the country of the requester. ISO copyright office Case postale 56 • CH-1211 Geneva 20 Tel.
+ 41 22 749 01 11 Fax
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copyright@iso.org Web
www.iso.org Reproduction may be subject to royalty payments or a licensing agreement. Violators may be prosecuted.
ii © ISO 2008 – All rights reserved
ISO/FDIS 15819:2008(E) © ISO 2008 – All rights reserved iiiForeword 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 15819 was prepared by Technical Committee ISO/TC 217, Cosmetics.
ISO/FDIS 15819:2008(E) iv © ISO 2008 – All rights reserved Introduction Human exposure to N-nitrosamines can occur through diverse sources such as environment, food or personal care products. As a result of their perceived carcinogenic potential on several animal species, minimization of exposure to N-nitrosamines is recognised as important to the preservation of human health. Among N-nitrosamines, N-nitrosodiethanolamine (NDELA) has been recognised as a potential contaminant of cosmetics. In this context, several analytical methods have been developed to detect and determine its presence in cosmetics – such as gas chromatography/thermal energy analysis, high performance liquid chromatography (HPLC) coupled either with photolysis and colorimetric quantification or with mass spectrometry (MS) determination. This latter method uses advanced technology to ensure the maximum specificity towards NDELA, to minimize the risk of artifactual formation of the analyte of interest and to allow precise quantification. This analytical method uses high performance liquid chromatography coupled with mass spectrometry to separate and detect trace levels of NDELA from a cosmetic ingredient or product matrix with maximum specificity for NDELA.
FINAL DRAFT INTERNATIONAL STANDARD ISO/FDIS 15819:2008(E) © ISO 2008 – All rights reserved 1Cosmetics — Analytical methods — Nitrosamines: Detection and determination of N-nitrosodiethanolamine (NDELA) in cosmetics by HPLC-MS 1 Scope This International Standard describes a method for the detection and quantification of NDELA in cosmetics and raw materials used in cosmetics. This method is not applicable to the detection and/or quantification of nitrosamines other than NDELA nor to the detection and/or quantification of NDELA in products other than cosmetics or raw materials used in cosmetics. If a product has a possibility of either NDELA contamination from ingredients or NDELA formation by the composition of ingredients, the method shall be applied for quantitative determination of NDELA. Accordingly the method would not be applied in routine testing of cosmetic products. Because of the large variety of cosmetic products within this field of application, this method might need to be adapted for certain matrices. Therefore, International Standards dedicated to alternative methods for testing nitrosamines in cosmetic products are being developed separately. Other methods can be employed provided that they are verified as to their detection of NDELA and validated in terms of recovery and quantification of the analyte. 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 3696:1987, Water for analytical laboratory use — Specification and test methods 3 Principle Extraction of the nitrosamine NDELA in cosmetic samples is carried out with water in the presence of deuterated d8-NDELA used as internal standard. Clean-up is performed either using solid phase extraction (SPE clean-up, see 6.3.1) with a C18 cartridge or liquid–liquid extraction using dichloromethane (DCM clean-up, see 6.3.2) when the samples are not dispersible in water. The extracts are analysed by HPLC-MS-MS (high performance liquid chromatography coupled with tandem mass spectrometric detection). NDELA quantification is done by comparing the ratio of the major fragment ions of NDELA and d8-NDELA with the calibration curve. Confirmation of the presence of NDELA is carried out by using the molecular ion and two diagnostic ions.
ISO/FDIS 15819:2008(E) 2 © ISO 2008 – All rights reserved 4 Reagents During the analysis, unless otherwise stated, use only reagents of recognized analytical grade and only distilled water or water of grade 1 in accordance with ISO 3696:1987. Solvent shall be of quality for HPLC analysis. 4.1 Methanol (MeOH), HPLC grade. 4.2 Ethanol (EtOH), HPLC grade. 4.3 Dichloromethane, HPLC grade. 4.4 N-nitrosodiethanolamine, with known purity greater than 95 %. 4.5 d8-N-nitrosodiethanolamine, with known purity greater than 95 %. 4.6 Ammonium acetate (NH4Ac), analytical grade. 4.7 1 mol/l ammonium acetate solution, formed by dissolving 77,08 g of NH4Ac in 1,0 l water. 4.8 Eluent A: 2 mmol NH4Ac in water, formed by taking 2 ml of 1 mol/l NH4Ac (4.7) and making up to 1 l with water. 4.9 Eluent B: 2 mmol NH4Ac in 90 % MeOH/water, formed by taking 2 ml of 1 mol/l NH4Ac (4.7) and adding 900 ml MeOH and 98 ml water. 5 Apparatus Use standard laboratory glassware and equipment, with the addition of: 5.1 Vortex mixer. 5.2 Sample processing station, in SPE application (such as Vacmaster®1) sample processing station, IST). 5.3 Centrifuge, capable of reaching not less than 20 000 G. 5.4 Solid phase extraction columns, e.g. Bakerbond®1) C18 – 6 ml, 500 mg reversed phase octadecylsilane bonded to silicagel, 40 APD, 60 Å. 5.5 HPLC-MS-MS equipment consisting of: 5.5.1 High performance
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