Cheese, cheese rind and processed cheese - Determination of natamycin content - Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese (ISO 9233-2:2007 including Amd 1:2012)

This part of ISO 9233|IDF 140 specifies a method for the determination of natamycin mass fraction in cheese, cheese rind and processed cheese of above 0,5 mg/kg and of the surface-area-related natamycin mass in cheese rind of above 0,03 mg/dm2.

Käse, Käserinde und Schmelzkäse - Bestimmung des Natamycingehalts - Teil 2: Verfahren mit Hochleistungs-Flüssigchromatographie für Käse, Käserinde und Schmelzkäse (ISO 9233-2:2007 + Amd 1:2012)

Der vorliegende Teil von ISO 9233|IDF 140 legt ein Verfahren für die Bestimmung des Natamycin-Massen-anteils oberhalb von 0,5 mg/kg in Käse, Käserinde und Schmelzkäse sowie der oberflächenbezogenen Masse von Natamycin oberhalb von 0,03 mg/dm2 in Käserinde fest.

Fromage, croûte de fromage et fromages fondus - Détermination de la teneur en natamycine - Partie 2: Méthode par chromatographie liquide à haute performance pour fromage, croûte de fromage et fromages fondus (ISO 9233-2:2007, Amd 1:2012 inclus)

L'ISO 9233-2|FIL 140-2:2007 spécifie une méthode de détermination de la teneur en natamycine présente dans le fromage, la croûte de fromage et les fromages fondus supérieure à 0,5 mg/kg et de la masse, exprimée par rapport à la surface, de natamycine de la croûte de fromage supérieure à 0,03 mg/dm2.

Sir, skorja sira in topljeni sir - Določevanje natamicina - 2. del: Metoda tekočinske kromatografije visoke ločljivosti za sir, skorjo sira in topljeni sir (ISO 9233-2:2007, vključno z Amd 1:2012)

Ta del ISO 9233|IDF 140 določa metodo za določevanje natamicina z masnim deležem nad 0,5 mg/kg v siru, skorji sira in topljenemu siru ter mase natamicina na površini skorje sira nad 0,03 mg/dm2.

General Information

Status
Withdrawn
Public Enquiry End Date
14-Mar-2013
Publication Date
06-Aug-2013
Withdrawal Date
12-Jun-2018
Current Stage
9900 - Withdrawal (Adopted Project)
Start Date
13-Jun-2018
Due Date
06-Jul-2018
Completion Date
13-Jun-2018

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Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 9233-2:2013
01-september-2013
6LUVNRUMDVLUDLQWRSOMHQLVLU'RORþHYDQMHQDWDPLFLQDGHO0HWRGDWHNRþLQVNH
NURPDWRJUDILMHYLVRNHORþOMLYRVWL]DVLUVNRUMRVLUDLQWRSOMHQLVLU ,62
YNOMXþQR]$PG
Cheese, cheese rind and processed cheese - Determination of natamycin content - Part
2: High-performance liquid chromatographic method for cheese, cheese rind and
processed cheese (ISO 9233-2:2007 including Amd 1:2012)
Käse, Käserinde und Schmelzkäse - Bestimmung des Natamycingehalts - Teil 2:
Verfahren mit Hochleistungs-Flüssigchromatographie für Käse, Käserinde und
Schmelzkäse (ISO 9233-2:2007 + Amd 1:2012)
Fromage, croûte de fromage et fromages fondus - Détermination de la teneur en
natamycine - Partie 2: Méthode par chromatographie liquide à haute performance pour
fromage, croûte de fromage et fromages fondus (ISO 9233-2:2007, Amd 1:2012 inclus)
Ta slovenski standard je istoveten z: EN ISO 9233-2:2013
ICS:
67.100.30 Sir Cheese
SIST EN ISO 9233-2:2013 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------

SIST EN ISO 9233-2:2013

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SIST EN ISO 9233-2:2013


EUROPEAN STANDARD
EN ISO 9233-2

NORME EUROPÉENNE

EUROPÄISCHE NORM
May 2013
ICS 67.100.30
English Version
Cheese, cheese rind and processed cheese - Determination of
natamycin content - Part 2: High-performance liquid
chromatographic method for cheese, cheese rind and processed
cheese (ISO 9233-2:2007 including Amd 1:2012)
Fromage, croûte de fromage et fromages fondus - Käse, Käserinde und Schmelzkäse - Bestimmung des
Détermination de la teneur en natamycine - Partie 2: Natamycingehalts - Teil 2: Verfahren mit Hochleistungs-
Méthode par chromatographie liquide à haute performance Flüssigchromatographie für Käse, Käserinde und
pour fromage, croûte de fromage et fromages fondus (ISO Schmelzkäse (ISO 9233-2:2007 einschließlich Amd
9233-2:2007, Amd 1:2012 inclus) 1:2012)
This European Standard was approved by CEN on 16 May 2013.

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, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 9233-2:2013: E
worldwide for CEN national Members.

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SIST EN ISO 9233-2:2013
EN ISO 9233-2:2013 (E)
Contents Page
Foreword .3

2

---------------------- Page: 4 ----------------------

SIST EN ISO 9233-2:2013
EN ISO 9233-2:2013 (E)
Foreword
The text of ISO 9233-2:2007 including Amd 1:2012 has been prepared by Technical Committee ISO/TC 34
“Food products” of the International Organization for Standardization (ISO) and has been taken over as EN
ISO 9233-2:2013 by Technical Committee CEN/TC 302 “Milk and milk products - Methods of sampling and
analysis” the secretariat of which is held by NEN.
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 November 2013, and conflicting national standards shall be withdrawn
at the latest by November 2013.
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, 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, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 9233-2:2007 including Amd 1:2012 has been approved by CEN as EN ISO 9233-2:2013
without any modification.

3

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SIST EN ISO 9233-2:2013

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SIST EN ISO 9233-2:2013


INTERNATIONAL ISO
STANDARD 9233-2
IDF
140-2
First edition
2007-12-15

Cheese, cheese rind and processed
cheese — Determination of natamycin
content —
Part 2:
High-performance liquid
chromatographic method for cheese,
cheese rind and processed cheese
Fromage, croûte de fromage et fromages fondus — Détermination de la
teneur en natamycine —
Partie 2: Méthode par chromatographie liquide à haute performance
pour fromage, croûte de fromage et fromages fondus




Reference numbers
ISO 9233-2:2007(E)
IDF 140-2:2007(E)
©
ISO and IDF 2007

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SIST EN ISO 9233-2:2013
ISO 9233-2:2007(E)
IDF 140-2:2007(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. 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 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
and IDF national committees. In the unlikely event that a problem relating to it is found, please inform the ISO Central Secretariat at the
address given below.


COPYRIGHT PROTECTED DOCUMENT


©  ISO and IDF 2007
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 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. + 32 2 733 98 88
Fax + 41 22 749 09 47 Fax + 32 2 733 04 13
E-mail copyright@iso.org E-mail info@fil-idf.org
Web www.iso.org Web www.fil-idf.org
Published in Switzerland

ii © ISO and IDF 2007 – All rights reserved

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SIST EN ISO 9233-2:2013
ISO 9233-2:2007(E)
IDF 140-2:2007(E)
Contents Page
Foreword. iv
Foreword. v
1 Scope . 1
2 Terms and definitions. 1
3 Principle. 1
4 Reagents. 2
5 Apparatus . 2
6 Sampling. 3
7 Preparation of test sample. 3
7.1 Cheese rind . 3
7.2 Cheese interior and processed cheese. 4
8 Procedure . 4
8.1 Test portion . 4
8.2 Preparation of test solution . 4
8.3 Determination. 5
9 Calculation and expression of results. 6
9.1 Calculation of natamycin mass fraction. 6
9.2 Calculation of surface-area-related natamycin mass . 7
9.3 Correction of results. 7
9.4 Expression of results . 7
10 Precision. 7
10.1 Interlaboratory tests . 7
10.2 Repeatability. 7
10.3 Reproducibility. 7
11 Test report . 8
Annex A (informative) Examples . 9
Annex B (informative) Results of interlaboratory trial. 11
Bibliography . 12

© ISO and IDF 2007 – All rights reserved iii

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SIST EN ISO 9233-2:2013
ISO 9233-2:2007(E)
IDF 140-2:2007(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 9233-2|IDF 140-2 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 5,
Milk and milk products and the International Dairy Federation (IDF) and is being published jointly by ISO and
IDF.
This first edition of ISO 9233-2|IDF 140-2, together with ISO 9233-1|IDF 140-1, cancels and replaces the first
edition of ISO 9233:1991, which has been technically revised.
ISO 9233|IDF 140 consists of the following parts, under the general title Cheese, cheese rind and processed
cheese — Determination of natamycin content:
⎯ Part 1: Molecular absorption spectrometric method for cheese rind
⎯ Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese


iv © ISO and IDF 2007 – All rights reserved

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SIST EN ISO 9233-2:2013
ISO 9233-2:2007(E)
IDF 140-2:2007(E)
Foreword
IDF (the International Dairy Federation) is a worldwide federation of the dairy sector with a National
Committee in every member country. Every National Committee has the right to be represented on the IDF
Standing Committees carrying out the technical work. IDF collaborates with ISO in the development of
standard methods of analysis and sampling for milk and milk products.
Draft International Standards adopted by the Action Teams and Standing Committees are circulated to the
National Committees for voting. Publication as an International Standard requires approval by at least 50% of
IDF National Committees casting a vote.
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 for identifying any or all such patent rights.
ISO 9233-2|IDF 140-2 was prepared by the International Dairy Federation (IDF) and Technical Committee
ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products and is being published jointly by IDF
and ISO.
All work was carried out by the Joint ISO-IDF Action Team on Selected food additives and vitamins of the
Standing Committee on Analytical methods for additives and contaminants under the aegis of its project
leader, Mr. M. Carl (DE).
This first edition of ISO 9233-2|IDF 140-2, together with ISO 9233-1|IDF 140-1, cancels and replaces the first
edition of IDF 140A:1992, which has been technically revised.
ISO 9233|IDF 140 consists of the following parts, under the general title Cheese, cheese rind and processed
cheese — Determination of natamycin content:
⎯ Part 1: Molecular absorption spectrometric method for cheese rind
⎯ Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese


© ISO and IDF 2007 – All rights reserved v

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SIST EN ISO 9233-2:2013

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SIST EN ISO 9233-2:2013
ISO 9233-2:2007(E)
INTERNATIONAL STANDARD
IDF 140-2:2007(E)

Cheese, cheese rind and processed cheese — Determination
of natamycin content —
Part 2:
High-performance liquid chromatographic method for cheese,
cheese rind and processed cheese
1 Scope
This part of ISO 9233|IDF 140 specifies a method for the determination of natamycin mass fraction in cheese,
cheese rind and processed cheese of above 0,5 mg/kg and of the surface-area-related natamycin mass in
2
cheese rind of above 0,03 mg/dm .
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1
natamycin content
mass fraction of substances determined by the procedure specified in this part of ISO 9233|IDF 140
NOTE The natamycin content is expressed in milligrams per kilogram.
2.2
surface-area-related natamycin mass in cheese rind
surface-area-related mass of substances determined by the procedure specified in this part of
ISO 9233|IDF 140
NOTE The surface-area-related natamycin mass is expressed in milligrams of natamycin per square decimetre of
cheese rind.
2.3
cheese rind
outer layer of the cheese of thickness 5 mm, excluding the coating layer, if present.
3 Principle
A known quantity of sample is extracted with methanol. The extract is diluted with water followed by cooling to
between −15 °C and −20 °C to precipitate most of the fat, followed by filtration. The natamycin content or
surface-area-related natamycin mass is determined in the filtrate (after concentration, if necessary) by high-
performance liquid chromatography (HPLC).
© ISO and IDF 2007 – All rights reserved 1

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SIST EN ISO 9233-2:2013
ISO 9233-2:2007(E)
IDF 140-2:2007(E)
4 Reagents
Use only reagents of recognized analytical grade, unless otherwise specified, and only distilled or
demineralized water or water of equivalent purity.
4.1 Methanol (CH OH).
3
4.2 Methanol, aqueous solution.
Mix 2 volumes of methanol (4.1) with 1 volume of water.
4.3 Natamycin standard solutions.
4.3.1 Natamycin standard stock solution, of concentration 500 mg/l.
Immediately before use, dissolve in methanol (4.1) a quantity of a natamycin preparation of known natamycin
content, corresponding to 50 mg of pure natamycin (C H NO ), in a 100 ml one-mark volumetric flask (5.1).
33 47 13
Make up to the mark with water and mix.
4.3.2 Natamycin standard working solution, of concentration 5 mg/l.
Pipette 5,0 ml of the natamycin standard stock solution (4.3.1) in a 50 ml one-mark volumetric flask (5.1).
Dilute to the mark with aqueous methanol (4.2) and mix.
Pipette 5,0 ml of the thus diluted solution into another 50 ml one-mark volumetric flask (5.1). Dilute to the mark
with aqueous methanol (4.2) and mix. The concentration of this natamycin standard working solution is
5 µg/ml.
This concentration shall be close to that of the test solution measured in 8.3.3. Adjust the standard working
dilution by pipetting and diluting another quantity, if required.
4.4 Acetic acid (CH CO H), glacial.
3 2
5 Apparatus
Usual laboratory equipment and, in particular, the following.
5.1 One-mark volumetric flasks, of capacities 50 ml and 100 ml.
5.2 Slicer or similar apparatus, capable of cutting cheese portions of thickness 5 mm and of width about
30 mm (Figure A.1 shows an example).
5.3 Fine slicer, capable of cutting thin cheese slices of maximum thickness 1 mm (Figure A.2 shows an
example).
5.4 Grinder or blender.
5.5 Sharp knife, capable of cutting cheese slices into small pieces.
5.6 Magnetic stirrer or shaking machine.
5.7 Conical flasks, of capacities 100 ml and 200 ml, made of coloured glass and fitted with ground-glass
stoppers.
5.8 Syringes, disposable, of capacity 10 ml.
2 © ISO and IDF 2007 – All rights reserved

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SIST EN ISO 9233-2:2013
ISO 9233-2:2007(E)
IDF 140-2:2007(E)
5.9 Membrane microfilters, of pore size 0,20 µm and 0,45 µm, resistant to attack by alcoholic solutions.
1)
5.10 Folded paper filters, fast speed, of diameter 150 mm. [e.g. S and S, No. 595 1/2 ].
5.11 Funnel, of diameter approximately 70 mm.
5.12 Freezer, capable of freezing at a temperature of between −15 °C and −20 °C.
1)
5.13 Extraction cartridges, to concentrate the filtered extract, if necessary [e.g. Sep-pack C18 or Waters
1)
No. 51910 ].
5.14 Liquid chromatograph, with UV detector, capable of measuring at 303 nm and equipped with a
recorder and/or integrator.
5.15 Analytical column, of length 150 mm, of internal diameter 4,6 mm, type C8, having a particle size of
1)
5 µm [e.g.
...

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Käse, Käserinde und Schmelzkäse - Bestimmung des Natamycingehalts - Teil 2: Verfahren mit Hochleistungs-Flüssigchromatographie für Käse, Käserinde und Schmelzkäse (ISO 9233-2:2007 + Amd 1:2012)Fromage, croûte de fromage et fromages fondus - Détermination de la teneur en natamycine - Partie 2: Méthode par chromatographie liquide à haute performance pour fromage, croûte de fromage et fromages fondus (ISO 9233-2:2007, Amd 1:2012 inclus)Cheese, cheese rind and processed cheese - Determination of natamycin content - Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese (ISO 9233-2:2007 including Amd 1:2012)67.100.30SirCheeseICS:Ta slovenski standard je istoveten z:FprEN ISO 9233-2kSIST FprEN ISO 9233-2:2013en01-februar-2013kSIST FprEN ISO 9233-2:2013SLOVENSKI
STANDARD



kSIST FprEN ISO 9233-2:2013



EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
FINAL DRAFT
FprEN ISO 9233-2
November 2012 ICS 67.100.30 English Version
Cheese, cheese rind and processed cheese - Determination of natamycin content - Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese (ISO 9233-2:2007 including Amd 1:2012)
Fromage, croûte de fromage et fromages fondus - Détermination de la teneur en natamycine - Partie 2: Méthode par chromatographie liquide à haute performance pour fromage, croûte de fromage et fromages fondus (ISO 9233-2:2007, Amd 1:2012 inclus)
Käse, Käserinde und Schmelzkäse - Bestimmung des Natamycingehalts - Teil 2: Verfahren mit Hochleistungs-Flüssigchromatographie für Käse, Käserinde und Schmelzkäse (ISO 9233-2:2007 + Amd 1:2012) This draft European Standard is submitted to CEN members for unique acceptance procedure. It has been drawn up by the Technical Committee CEN/TC 302.
If this draft becomes a European Standard, 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.
This draft European Standard was established by CEN 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, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are aware and to provide supporting documentation.
Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without notice and shall not be referred to as a European Standard.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre:
Avenue Marnix 17,
B-1000 Brussels © 2012 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. FprEN ISO 9233-2:2012: EkSIST FprEN ISO 9233-2:2013



FprEN ISO 9233-2:2012 (E) 2 Contents Page Foreword .3 kSIST FprEN ISO 9233-2:2013



FprEN ISO 9233-2:2012 (E) 3 Foreword The text of ISO 9233-2:2007 including Amd 1:2012 has been prepared by Technical Committee ISO/TC 34 “Food products” of the International Organization for Standardization (ISO) and has been taken over as FprEN ISO 9233-2:2012 by Technical Committee CEN/TC 302 “Milk and milk products - Methods of sampling and analysis” the secretariat of which is held by NEN. This document is currently submitted to the Unique Acceptance Procedure. Endorsement notice The text of ISO 9233-2:2007 including Amd 1:2012 has been approved by CEN as a FprEN ISO 9233-2:2012 without any modification.
kSIST FprEN ISO 9233-2:2013



kSIST FprEN ISO 9233-2:2013



Reference numbersISO 9233-2:2007(E)IDF 140-2:2007(E)© ISO and IDF 2007
INTERNATIONAL STANDARD ISO9233-2IDF140-2First edition2007-12-15Cheese, cheese rind and processed cheese — Determination of natamycin content — Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese Fromage, croûte de fromage et fromages fondus — Détermination de la teneur en natamycine — Partie 2: Méthode par chromatographie liquide à haute performance pour fromage, croûte de fromage et fromages fondus
kSIST FprEN ISO 9233-2:2013



ISO 9233-2:2007(E) IDF 140-2:2007(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. 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 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 and IDF national committees. In the unlikely event that a problem relating to it is found, please inform the ISO Central Secretariat at the address given below.
COPYRIGHT PROTECTED DOCUMENT
©
ISO and IDF 2007 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 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.
+ 32 2 733 98 88 Fax
+ 41 22 749 09 47 Fax
+ 32 2 733 04 13 E-mail
copyright@iso.org E-mail
info@fil-idf.org Web
www.iso.org Web
www.fil-idf.org Published in Switzerland
ii © ISO and IDF 2007 – All rights reserved
kSIST FprEN ISO 9233-2:2013



ISO 9233-2:2007(E) IDF 140-2:2007(E) © ISO and IDF 2007 – All rights reserved iiiContents Page Foreword.iv Foreword.v 1 Scope.1 2 Terms and definitions.1 3 Principle.1 4 Reagents.2 5 Apparatus.2 6 Sampling.3 7 Preparation of test sample.3 7.1 Cheese rind.3 7.2 Cheese interior and processed cheese.4 8 Procedure.4 8.1 Test portion.4 8.2 Preparation of test solution.4 8.3 Determination.5 9 Calculation and expression of results.6 9.1 Calculation of natamycin mass fraction.6 9.2 Calculation of surface-area-related natamycin mass.7 9.3 Correction of results.7 9.4 Expression of results.7 10 Precision.7 10.1 Interlaboratory tests.7 10.2 Repeatability.7 10.3 Reproducibility.7 11 Test report.8 Annex A (informative)
Examples.9 Annex B (informative)
Results of interlaboratory trial.11 Bibliography.12
kSIST FprEN ISO 9233-2:2013



ISO 9233-2:2007(E) IDF 140-2:2007(E) iv © ISO and IDF 2007 – All rights reserved 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 9233-2|IDF 140-2 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products and the International Dairy Federation (IDF) and is being published jointly by ISO and IDF. This first edition of ISO 9233-2|IDF 140-2, together with ISO 9233-1|IDF 140-1, cancels and replaces the first edition of ISO 9233:1991, which has been technically revised. ISO 9233|IDF 140 consists of the following parts, under the general title Cheese, cheese rind and processed cheese — Determination of natamycin content: ⎯ Part 1: Molecular absorption spectrometric method for cheese rind ⎯ Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese
kSIST FprEN ISO 9233-2:2013



ISO 9233-2:2007(E) IDF 140-2:2007(E) © ISO and IDF 2007 – All rights reserved vForeword IDF (the International Dairy Federation) is a worldwide federation of the dairy sector with a National Committee in every member country. Every National Committee has the right to be represented on the IDF Standing Committees carrying out the technical work. IDF collaborates with ISO in the development of standard methods of analysis and sampling for milk and milk products. Draft International Standards adopted by the Action Teams and Standing Committees are circulated to the National Committees for voting. Publication as an International Standard requires approval by at least 50% of IDF National Committees casting a vote. 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 for identifying any or all such patent rights. ISO 9233-2|IDF 140-2 was prepared by the International Dairy Federation (IDF) and Technical Committee ISO/TC 34, Food products, Subcommittee SC 5, Milk and milk products and is being published jointly by IDF and ISO. All work was carried out by the Joint ISO-IDF Action Team on Selected food additives and vitamins of the Standing Committee on Analytical methods for additives and contaminants under the aegis of its project leader, Mr. M. Carl (DE). This first edition of ISO 9233-2|IDF 140-2, together with ISO 9233-1|IDF 140-1, cancels and replaces the first edition of IDF 140A:1992, which has been technically revised. ISO 9233|IDF 140 consists of the following parts, under the general title Cheese, cheese rind and processed cheese — Determination of natamycin content: ⎯ Part 1: Molecular absorption spectrometric method for cheese rind ⎯ Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese
kSIST FprEN ISO 9233-2:2013



kSIST FprEN ISO 9233-2:2013



INTERNATIONAL STANDARD ISO 9233-2:2007(E)IDF 140-2:2007(E) © ISO and IDF 2007 – All rights reserved 1Cheese, cheese rind and processed cheese — Determination of natamycin content — Part 2: High-performance liquid chromatographic method for cheese, cheese rind and processed cheese 1 Scope This part of ISO 9233|IDF 140 specifies a method for the determination of natamycin mass fraction in cheese, cheese rind and processed cheese of above 0,5 mg/kg and of the surface-area-related natamycin mass in cheese rind of above 0,03 mg/dm2. 2 Terms and definitions For the purposes of this document, the following terms and definitions apply. 2.1 natamycin content mass fraction of substances determined by the procedure specified in this part of ISO 9233|IDF 140 NOTE The natamycin content is expressed in milligrams per kilogram. 2.2 surface-area-related natamycin mass in cheese rind surface-area-related mass of substances determined by the procedure specified in this part of ISO 9233|IDF 140 NOTE The surface-area-related natamycin mass is expressed in milligrams of natamycin per square decimetre of cheese rind. 2.3 cheese rind outer layer of the cheese of thickness 5 mm, excluding the coating layer, if present. 3 Principle A known quantity of sample is extracted with methanol. The extract is diluted with water followed by cooling to between −15 °C and −20 °C to precipitate most of the fat, followed by filtration. The natamycin content or surface-area-related natamycin mass is determined in the filtrate (after concentration, if necessary) by high-performance liquid chromatography (HPLC). kSIST FprEN ISO 9233-2:2013



ISO 9233-2:2007(E) IDF 140-2:2007(E) 2 © ISO and IDF 2007 – All rights reserved 4 Reagents Use only reagents of recognized analytical grade, unless otherwise specified, and only distilled or demineralized water or water of equivalent purity. 4.1 Methanol (CH3OH). 4.2 Methanol, aqueous solution. Mix 2 volumes of methanol (4.1) with 1 volume of water. 4.3 Natamycin standard solutions. 4.3.1 Natamycin standard stock solution, of concentration 500 mg/l. Immediately before use, dissolve in methanol (4.1) a quantity of a natamycin preparation of known natamycin content, corresponding to 50 mg of pure natamycin (C33H47NO13), in a 100 ml one-mark volumetric flask (5.1). Make up to the mark with water and mix. 4.3.2 Natamycin standard working solution, of concentration 5 mg/l. Pipette 5,0 ml of the natamycin standard stock solution (4.3.1) in a 50 ml one-mark volumetric flask (5.1). Dilute to the mark with aqueous methanol (4.2) and mix. Pipette 5,0 ml of the thus diluted solution into another 50 ml one-mark volumetric flask (5.1). Dilute to the mark with aqueous methanol (4.2) and mix. The concentration of this natamycin standard working solution is 5 µg/ml. This concentration shall be close to that of the test solution measured in 8.3.3. Adjust the standard working dilution by pipetting and diluting another quantity, if required. 4.4 Acetic acid (CH3CO2H), glacial. 5 Apparatus Usual laboratory equipment and, in particular, the following. 5.1 One-mark volumetric flasks, of capacities 50 ml and 100 ml. 5.2 Slicer or similar apparatus, capable of cutting cheese portions of thickness 5 mm and of width about 30 mm (Figure A.1 shows an example). 5.3 Fine slicer, capable of cutting thin cheese slices of maximum thickness 1 mm (Figure A.2 shows an example). 5.4 Grinder or blender. 5.5 Sharp knife, capable of cutting cheese slices into small pieces. 5.6 Magnetic stirrer or shaking machine. 5.7 Conical flasks, of capacities 100 ml and 200 ml, made of coloured glass and fitted with ground-glass stoppers. 5.8 Syringes, disposable, of capacity 10 ml. kSIST FprEN ISO 9233-2:2013



ISO 9233-2:2007(E) IDF 140-2:2007(E) © ISO and IDF 2007 – All rights reserved 35.9 Membrane microfilters, of pore size 0,20 µm and 0,45 µm, resistant to attack by alcoholic solutions. 5.10 Folded paper filters, fast speed, of diameter 150 mm. [e.g. S and S, No. 595 1/21)]. 5.11 Funnel, of diameter approximately 70 mm. 5.12 Freezer, capable of freezing at a temperature of between −15 °C and −20 °C. 5.13 Extraction cartridges, to concentrate the filtered extract, if necessary [e.g. Sep-pack C181) or Waters No. 519101)]. 5.14 Liquid chromatograph, with UV detector, capable of measuring at 303 nm and equipped with a recorder and/or integrator. 5.15 Analytical column, of length 150 mm, of internal diameter 4,6 mm, type C8, having a particle size of 5 µm [e.g. Lichrosorb RP81)]. 5.16 Guard column, of length 100 mm, of internal diameter 2,1 mm, type C8, having a particle size of 30 µm to 40 µm [e.g. Perisorb RP81)]. 5.17 Sample jar, of suitable capacity. 6 Sampling A representative sample should have been sent to the laboratory. It should not have been damaged or changed during transport or storage. Sampling is not part of the method specified in this part of ISO 9233|IDF 140. A recommended sampling method is given in ISO
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