1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationDD ISO/TS 15495:2010Milk, milk products and infant formulae Guidelines for the quantitative determination of melamine and cyanuric acid by LC-MS/MSDD ISO/TS 15495:2010 DRAFT FOR
2、DEVELOPMENTNational forewordThis Draft for Development is the UK implementation of ISO/TS 15495:2010.This publication is not to be regarded as a British Standard.It is being issued in the Draft for Development series of publicationsand is of a provisional nature. It should be applied on thisprovisio
3、nal basis, so that information and experience of its practicalapplication can be obtained.Comments arising from the use of this Draft for Developmentare requested so that UK experience can be reported to theinternational organization responsible for its conversion toan international standard. A revi
4、ew of this publication willbe initiated not later than 3 years after its publication by theinternational organization so that a decision can be taken on itsstatus. Notification of the start of the review period will be made inan announcement in the appropriate issue of Update Standards. According to
5、 the replies received by the end of the review period,the responsible BSI Committee will decide whether to support theconversion into an international Standard, to extend the life of theTechnical Specification or to withdraw it. Comments should be sentto the Secretary of the responsible BSI Technica
6、l Committee at BritishStandards House, 389 Chiswick High Road, London W4 4AL.The UK participation in its preparation was entrusted to TechnicalCommittee AW/5, Chemical analysis of milk and milk products. A list of organizations represented on this committee can beobtained on request to its secretary
7、.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. BSI 2011 ISBN 978 0 580 70431 4 ICS 67.100.01 Compliance with a British Standard cannot confer immunity fromlegal obligations.This Draft for Development was publ
8、ished under the authority ofthe Standards Policy and Strategy Committee on 31 March 2011. Amendments issued since publicationDate Text affectedDD ISO/TS 15495:2010Reference numbers ISO/TS 15495:2010(E)IDF/RM 230:2010(E)ISO and IDF 2010TECHNICAL SPECIFICATION ISO/TS 15495IDF/RM 230First edition 2010-
9、11-01Milk, milk products and infant formulae Guidelines for the quantitative determination of melamine and cyanuric acid by LC-MS/MS Lait, produits laitiers et formules infantiles Lignes directrices pour la dtermination quantitative de la mlamine et de lacide cyanurique par CL-SM/SM DD ISO/TS 15495:
10、2010 ISO/TS 15495:2010(E) IDF/RM 230:2010(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobes 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 perfo
11、rming the editing. In downloading this file, parties accept therein the responsibility of not infringing Adobes licensing policy. Neither the ISO Central Secretariat nor the IDF accepts any liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products u
12、sed 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 and IDF national committees. In the unlikely event that a problem rel
13、ating to it is found, please inform the ISO Central Secretariat at the address given below. COPYRIGHT PROTECTED DOCUMENT ISO and IDF 2010 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,
14、including photocopying and microfilm, without permission in writing from either ISO or IDF at the respective address below. ISO copyright office International Dairy Federation Case postale 56 CH-1211 Geneva 20 Diamant Building Boulevard Auguste Reyers 80 B-1030 Brussels Tel. + 41 22 749 01 11 Tel. +
15、 32 2 733 98 88 Fax + 41 22 749 09 47 Fax + 32 2 733 04 13 E-mail copyrightiso.org E-mail infofil-idf.org Web www.iso.org Web www.fil-idf.org Published in Switzerland ii ISO and IDF 2010 All rights reservedDD ISO/TS 15495:2010ISO/TS 15495:2010(E) IDF/RM 230:2010(E) ISO and IDF 2010 All rights reserv
16、ed iiiContents Page Foreword iv Foreword .v 1 Scope1 2 Normative references1 3 Terms and definitions .1 4 Principle .2 5 Sampling 2 6 Preparation of test sample .2 7 Procedure.2 7.1 LC-MS/MS analysis Chromatography.2 7.2 LC-MS/MS analysis Mass spectrometry.3 8 Performance criteria3 8.1 General .3 8.
17、2 Minimum required sensitivity.3 8.3 Trueness and recovery .3 8.4 Repeatability 3 8.5 Within-laboratory reproducibility.3 9 Test report4 Annex A (informative) Example A Cow milk and milk-based infant formula Simultaneous quantitative determination of melamine and cyanuric acid by liquid chromatograp
18、hy elec-trospray ionization tandem mass spectrometry .5 Annex B (informative) Example B Quantitative analysis of melamine and cyanuric acid in milk-based infant formula and cow milk by liquid chromatography using linear ion-trap mass spec-trometry19 Bibliography29 DD ISO/TS 15495:2010 ISO/TS 15495:2
19、010(E) IDF/RM 230:2010(E) iv ISO and IDF 2010 All rights reservedForeword 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 c
20、ommittees. 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 t
21、he 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 Internat
22、ional 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. In other circumstances, particularly when there is an urgent market requirement for suc
23、h documents, a technical committee may decide to publish other types of document: an ISO Publicly Available Specification (ISO/PAS) represents an agreement between technical experts in an ISO working group and is accepted for publication if it is approved by more than 50 % of the members of the pare
24、nt committee casting a vote; an ISO Technical Specification (ISO/TS) represents an agreement between the members of a technical commit-tee and is accepted for publication if it is approved by 2/3 of the members of the committee casting a vote. An ISO/PAS or ISO/TS is reviewed after three years in or
25、der to decide whether it will be confirmed for a further three years, revised to become an International Standard, or withdrawn. If the ISO/PAS or ISO/TS is confirmed, it is reviewed again after a further three years, at which time it must either be transformed into an International Standard or be w
26、ithdrawn. 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/TS 15495IDF/RM 230 was prepared by Technical Committee ISO/TC 34, Food products, Subcom
27、mittee SC 5, Milk and milk products, and the International Dairy Federation (IDF). It is being published jointly by ISO and IDF. DD ISO/TS 15495:2010ISO/TS 15495:2010(E) IDF/RM 230:2010(E) ISO and IDF 2010 All rights reserved vForeword IDF (the International Dairy Federation) is a non-profit organiz
28、ation representing the dairy sector worldwide. IDF membership comprises National Committees in every member country as well as regional dairy associations having signed a formal agreement on cooperation with IDF. All members of IDF have the right to be represented on the IDF Standing Committees carr
29、ying out the technical work. IDF collaborates with ISO in the development of standard methods of analysis and sampling for milk and milk products. The main task of Standing Committees is to prepare International Standards. Draft International Standards adopted by the Standing Committees are circulat
30、ed to the National Committees for endorsement prior to publication as an International Standard. Publication as an International Standard requires approval by at least 50 % of IDF National Committees casting a vote. In other circumstances, particularly when there is an urgent market requirement for
31、such documents, a Standing Committee may decide to publish an other type of normative document which is called by IDF: Reviewed method. Such a method represents an agreement between the members of a Standing Committee and is accepted for publication if it is approved by at least 50 % of the committe
32、e members casting a vote. A Reviewed method is equal to an ISO/PAS or ISO/TS and will, therefore, also be published jointly under ISO conditions. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. IDF shall not be held responsible fo
33、r identifying any or all such patent rights. ISO/TS 15495IDF/RM 230 was prepared by the International Dairy Federation (IDF) and Technical Committee ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products. It is being published jointly by IDF and ISO. All work was carried out by a Joint
34、Project Group on Determination of melamine of the Standing Committee on Analytical methods for additives and contaminants under the aegis of its project leaders, Dr S.E. Holroyd (NZ) and Dr T. Delatour (CH). DD ISO/TS 15495:2010DD ISO/TS 15495:2010TECHNICAL SPECIFICATION ISO/TS 15495:2010(E)IDF/RM 2
35、30:2010(E) ISO and IDF 2010 All rights reserved 1Milk, milk products and infant formulae Guidelines for the quantitative determination of melamine and cyanuric acid by LC-MS/MS 1 Scope This Technical Specification gives guidance for the quantitative determination of melamine and cyanuric acid conten
36、t in milk, powdered milk products, and infant formulae by electrospray ionization liquid chromatography tandem mass spectrometry (LC-MS/MS). NOTE Examples of LC-MS/MS methods are given in Annexes A and B. 2 Normative references The following referenced documents are indispensable for the application
37、 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 3534 (all parts), Statistics Vocabulary and symbols ISO 5725-1, Accuracy (trueness and precision) of measurement meth
38、ods and results Part 1: General principles and definitions 3 Terms and definitions For the purposes of this document, the terms and definitions given in ISO 3534, ISO 5725-1, and the following apply. 3.1 melamine content mass fraction of substance determined by the procedures specified in this Techn
39、ical Specification NOTE The melamine content is expressed in milligrams per kilogram of product. 3.2 cyanuric acid content mass fraction of substance determined by the procedures specified in this Technical Specification NOTE The cyanuric content is expressed in milligrams per kilogram of product. 3
40、.3 maximum limit ML maximum content of either melamine or cyanuric acid permitted or acceptable in milk, powdered milk products or infant formulae NOTE The ML can either be a nationally or internationally stated limit or a defined level of concern. DD ISO/TS 15495:2010 ISO/TS 15495:2010(E) IDF/RM 23
41、0:2010(E) 2 ISO and IDF 2010 All rights reserved4 Principle The sample is made homogenous and optionally reconstituted in the case of powdered samples. A suitable solvent is added to the test sample to precipitate proteins from the matrix and to extract melamine and cyanuric acid. After centrifugati
42、on, an aliquot of the supernatant is analysed by LC-MS/MS. For the purposes of this Technical Specification, LC-MS/MS designates any method combining either high-performance liquid chromatography (HPLC) or ultra-performance liquid chromatography (UPLC), with either triple quadrupole or ion-trap mass
43、 spectrometric detection. Chromatographic separation is based on hydrophilic interaction liquid chromatography (HILIC) to ensure good separation of melamine and cyanuric acid. Ionization of the substance is accomplished by electrospray ionization (ESI) and detection operates in the selected reaction
44、 monitoring (SRM) mode. NOTE Other ionization techniques with sufficient performance can be used. Other mass analysers can be used as long as they comply with the performance criteria (identification points) in 2002/657/EC (Reference 8). Quantification of both melamine and cyanuric acid is based on
45、isotope dilution using stable isotope internal standards for both analytes. 5 Sampling Sampling is not part of the method specified in this Technical Specification. A recommended sampling method is given in ISO 707IDF 501. It is important the laboratory receive a truly representative sample which ha
46、s not been damaged or changed during transport or storage. 6 Preparation of test sample The mass of the test sample for analysis or optional reconstitution should be representative of the product and should be appropriate to allow effective extraction of the target analytes. Isotope-labelled standar
47、ds should be spiked directly to the dry or wet sample in the first step of analysis. Concentration of internal standards added should be of the same order of magnitude as that of interest for melamine and cyanuric acid. Extraction may be done with a minimum of 5 ml of extraction solvent per gram of
48、sample material, to give a final percentage mass fraction of organic solvent in the extraction mixture of at least 70 %. Extraction should take place for at least 5 min by either vigorous shaking or a combination of ultrasonification and vortexing. The sample should then be centrifuged under appropr
49、iate conditions and filtered through a syringe filter. Any alternative is acceptable as long as performance criteria and validation match the requirements specified in this Technical Specification. 7 Procedure 7.1 LC-MS/MS analysis Chromatography Perform the liquid chromatographic separation using a column dedicated to this purpose. The minimum acceptable retention time for melamine and cyanuric acid shall be twice the retention time corresponding to the void volume of the column. The retention time in the sample ext
