SIST EN ISO 11816-2:2003

Milk and milk products - Determination of alkaline phosphatase activity - Part 2: Fluorometric method for cheese (ISO 11816-2:2003)

ISO 11816-2|IDF 155-2:2003 specifies a fluorometric method for the determination of alkaline phosphatase activity in cheese.
This method is also applicable to soft cheeses and semi-hard cheeses for distinguishing raw milk cheeses from cheese produced with pasteurized milk, provided that the mould is only on the surface and not also in the inner part (e.g. blue-veined cheeses). This method can also be used to check the proper pasteurization of cheese or its raw material.
In large hard cheeses where the whey curd mixture is scalded at temperatures above 50 degrees Celsius, high temperatures remain for a relatively long time. This is especially the case in the centre of these cheeses, thus promoting phosphatase inactivation. To distinguish, therefore, between hard cheeses from raw milk and hard cheeses from pasteurized milk is difficult and often even impossible by this method.

Milch und Milchprodukte - Bestimmung der Aktivität der alkalischen Phosphatase - Teil 2: Fluormetrisches Verfahren für Käse (ISO 11816-2:2003)

Dieser Teil von ISO 11816 | IDF 155 legt ein fluorimetrisches Verfahren für die Bestimmung der Aktivität der alkalischen Phosphatase in Käse fest.
Dieses Verfahren gilt für Weichkäse und Schnittkäse, um Rohmilchkäse von Käse zu unterscheiden, der mit pasteurisierter Milch hergestellt wurde. Das Verfahren setzt voraus, dass Schimmel nur an der Käseoberfläche und nicht auch im Innern des Käses vorhanden ist (z. B. Gorgonzola). Dieses Verfahren kann auch für die Überprüfung der Pasteurisierung von Käse oder Rohmilch angewandt werden.
In großen Hartkäselaiben, bei denen der Bruch auf Temperaturen von über 50 °C eingestellt wird, bleiben diese hohen Temperaturen besonders in der Mitte der Käselaibe für relativ lange Zeit bestehen. Deshalb erfordert die Unterscheidung zwischen Hartkäse aus Rohmilch und solchem aus pasteurisierter Milch mit diesem Verfahren eine spezifische Probenahmetechnik für Käse (siehe Abschnitt 7).

Lait et produits laitiers - Détermination de l'activité de la phosphatase alcaline - Partie 2: Méthode fluorométrique pour le fromage (ISO 11816-2:2003)

L'ISO 11816-2 | IDF 155-2:2003 spécifie une méthode fluorimétrique de détermination de l'activité de la phosphatase alcaline dans le fromage.
Cette méthode est également applicable aux fromages à pâte molle et demi-dure et permet de faire la distinction entre les fromages au lait cru et les fromages au lait pasteurisé, à condition que la moisissure ne soit présente que sur la surface et ne soit pas propagée à l'intérieur comme dans le cas des fromages veinés de bleu, par exemple. Cette méthode peut également être employée pour vérifier la validité de la pasteurisation du fromage ou de la matière première.
Les gros fromages à pâte dure où le mélange de caillé et de lactosérum est chauffé à des températures supérieures à 50 °C conservent des températures élevées pendant une durée relativement longue. Cela est notamment le cas en leurs centres, ce qui produit l'inactivation de la phosphatase. Pour faire la distinction entre les fromages à pâte dure au lait cru et ceux au lait pasteurisé, cette méthode nécessite une technique spécifique d'échantillonnage pour le fromage.

Mleko in mlečni proizvodi - Določevanje aktivnosti alkalne fosfataze s fluorometrijsko metodo - 2. del: Metoda za sir (ISO 11816-2:2003)

General Information

• Status: Withdrawn
• Publication Date: 31-Mar-2003
• Withdrawal Date: 18-Oct-2016
• ICS: 67.100.30 - Cheese
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 9900 - Withdrawal (Adopted Project)
• Start Date: 19-Oct-2016
• Due Date: 11-Nov-2016
• Completion Date: 19-Oct-2016
• Ref Project: EN ISO 11816-2:2003 - Milk and milk products - Determination of alkaline phosphatase activity - Part 2: Fluorometric method for cheese (ISO 11816-2:2003)

Relations

• Replaced By: SIST EN ISO 11816-2:2016 - Milk and milk products - Determination of alkaline phosphatase activity - Part 2: Fluorometric method for cheese (ISO 11816-2:2016)
• Effective Date: 01-Nov-2016
• Replaced By: SIST EN ISO 11816-2:2016 - Milk and milk products - Determination of alkaline phosphatase activity - Part 2: Fluorometric method for cheese (ISO 11816-2:2016)
• Effective Date: 08-Jun-2022

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 11816-2:2003
01-april-2003
0OHNRLQPOHþQLSURL]YRGL'RORþHYDQMHDNWLYQRVWLDONDOQHIRVIDWD]HV
IOXRURPHWULMVNRPHWRGRGHO0HWRGD]DVLU,62
Milk and milk products - Determination of alkaline phosphatase activity - Part 2:
Fluorometric method for cheese (ISO 11816-2:2003)
Milch und Milchprodukte - Bestimmung der Aktivität der alkalischen Phosphatase - Teil 2:
Fluormetrisches Verfahren für Käse (ISO 11816-2:2003)
Lait et produits laitiers - Détermination de l'activité de la phosphatase alcaline - Partie 2:
Méthode fluorométrique pour le fromage (ISO 11816-2:2003)
Ta slovenski standard je istoveten z: EN ISO 11816-2:2003
ICS:
67.100.30 Sir Cheese
SIST EN ISO 11816-2:2003 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 11816-2:2003

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SIST EN ISO 11816-2:2003
EUROPEAN STANDARD
EN ISO 11816-2
NORME EUROPÉENNE
EUROPÄISCHE NORM
February 2003
ICS 67.100.30
English version
Milk and milk products - Determination of alkaline phosphatase
activity - Part 2: Fluorometric method for cheese (ISO 11816-
2:2003)
Lait et produits laitiers - Détermination de l'activité de la Milch und Milchprodukte - Bestimmung der Aktivität der
phosphatase alcaline - Partie 2: Méthode fluorométrique alkalischen Phosphatase - Teil 2: Fluormetrisches
pour le fromage (ISO 11816-2:2003) Verfahren für Käse (ISO 11816-2:2003)
This European Standard was approved by CEN on 18 December 2002.
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 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 Management Centre has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Slovakia, Spain, Sweden, Switzerland and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36  B-1050 Brussels
© 2003 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 11816-2:2003 E
worldwide for CEN national Members.

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SIST EN ISO 11816-2:2003
EN ISO 11816-2:2003 (E)
CORRECTED 2003-03-26
Foreword
This document (EN ISO 11816-2:2003) has been prepared by Technical Committee ISO/TC 34
"Agricultural food products" in collaboration with 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 August 2003, and conflicting national
standards shall be withdrawn at the latest by August 2003.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of
the following countries are bound to implement this European Standard: Austria, Belgium, Czech
Republic, Denmark, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy,
Luxembourg, Malta, Netherlands, Norway, Portugal, Slovakia, Spain, Sweden, Switzerland and
the United Kingdom.
Endorsement notice
The text of ISO 11816-2:2003 has been approved by CEN as EN ISO 11816-2:2003 without any
modifications.
NOTE Normative references to International Standards are listed in Annex ZA (normative).
2

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SIST EN ISO 11816-2:2003
EN ISO 11816-2:2003 (E)
Annex ZA
(normative)
Normative references to international publications
with their relevant European publications
This European Standard incorporates by dated or undated reference, provisions from other
publications. These normative references are cited at the appropriate places in the text and the
publications are listed hereafter. For dated references, subsequent amendments to or revisions of
any of these publications apply to this European Standard only when incorporated in it by
amendment or revision. For undated references the latest edition of the publication referred to
applies (including amendments).
NOTE Where an International Publication has been modified by common modifications, indicated
by (mod.), the relevant EN/HD applies.
Publication Year Title EN Year
ISO 11816-1 1997 Milk and milk products - EN ISO 11816-1 2000
Determination of alkaline
phosphatase activity using a
fluorimetric method - Part 1:
Milk and milk-based drinks
3

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SIST EN ISO 11816-2:2003

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SIST EN ISO 11816-2:2003



INTERNATIONAL ISO
STANDARD 11816-2
IDF
155-2
First edition
2003-02-01


Milk and milk products — Determination
of alkaline phosphatase activity —
Part 2:
Fluorometric method for cheese
Lait et produits laitiers — Détermination de l'activité de la phosphatase
alcaline —
Partie 2: Méthode fluorométrique pour le fromage




Reference numbers
ISO 11816-2:2003(E)
IDF 155-2:2003(E)
©
ISO and IDF 2003

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SIST EN ISO 11816-2:2003
ISO 11816-2:2003(E)
IDF 155-2:2003(E)
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©  ISO and IDF 2003
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
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Published in Switzerland

ii © ISO and IDF 2003 – All rights reserved

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SIST EN ISO 11816-2:2003
ISO 11816-2:2003(E)
IDF 155-2:2003(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 part of ISO 11816|IDF 155 may be the
subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 11816-2IDF 155-2 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee
SC 5, Milk and milk products, and the International Dairy Federation (IDF), in collaboration with AOAC
International. It is being published jointly by ISO and IDF and separately by AOAC International.
ISO 11816IDF 155 consists of the following parts, under the general title Milk and milk products —
Determination of alkaline phosphatase activity:
 Part 1: Milk and milk-based drinks
 Part 2: Fluorometric method for cheese
 Part 3: Enzyme photo-activated system (EPAS) method for milk and milk-based products.
The present title of part 1 of ISO 11816 (IDF 155) is Milk and milk products — Determination of alkaline
phosphatase activity using a fluorometric method — Part 1: Milk and milk-based drinks. This title will be
changed to the title listed above when this part is revised.
© ISO and IDF 2003 – All rights reserved iii

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SIST EN ISO 11816-2:2003
ISO 11816-2:2003(E)
IDF 155-2:2003(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 and AOAC International 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.
ISO 11816-2IDF 155-2 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee
SC 5, Milk and milk products, and the International Dairy Federation (IDF), in collaboration with AOAC
International. It is being published jointly by ISO and IDF and separately by AOAC International.
All work was carried out by the Joint ISO/IDF/AOAC Action Team, Characterization of milk and milk products
according to heat treatment, of the Standing Committee on Minor components and characterization of physical
properties, under the aegis of its project leader, Mrs E. Lechner (DE).
ISO 11816IDF 155 consists of the following parts, under the general title Milk and milk products —
Determination of alkaline phosphatase activity:
 Part 1: Fluorometric method for milk and milk-based drinks
 Part 2: Fluorometric method for cheese
 Part 3: Enzyme photo-activated system (EPAS) method for milk and milk-based products
The present title of part 1 of ISO 11816 (IDF 155) is Milk and milk products — Determination of alkaline
phosphatase activity using a fluorometric method — Part 1: Milk and milk-based drinks. This title will be
changed to the title listed above when this part is revised.

iv © ISO and IDF 2003 – All rights reserved

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SIST EN ISO 11816-2:2003
ISO 11816-2:2003(E)
INTERNATIONAL STANDARD
IDF 155-2:2003(E)

Milk and milk products — Determination of alkaline
phosphatase activity —
Part 2:
Fluorometric method for cheese
1 Scope
This part of ISO 11816IDF 155 specifies a fluorometric method for the determination of alkaline phosphatase
activity in cheese.
This method is also applicable to soft cheeses and semi-hard cheeses for distinguishing raw milk cheeses
from cheese produced with pasteurized milk, provided that the mould is only on the surface and not also in the
inner part (e.g. blue-veined cheeses). This method can also be used to check the proper pasteurization of
cheese or its raw material.
In large hard cheeses where the whey curd mixture is scalded at temperatures above 50 °C, high
temperatures remain for a relatively long time. This is especially the case in the centre of these cheeses, thus
promoting phosphatase inactivation. To distinguish, therefore, between hard cheeses from raw milk and hard
cheeses from pasteurized milk using this method requires a specific sampling technique for the cheese (see
Clause 7).
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 11816-1, Milk and milk products — Determination of alkaline phosphatase activity using a fluorometric
method — Part 1: Milk and milk-based drinks
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
alkaline phosphatase activity
ALP
activity of the alkaline phosphatase present in the product, determined according to the procedure described
in this part of ISO 11816IDF 155
NOTE The alkaline phosphatase activity is expressed as milliunits of enzyme activity per gram.
3.2
unit of alkaline phosphatase activity
amount of alkaline phosphatase enzyme that catalyses the transformation of 1 µmol of substrate per minute
per gram of sample
© ISO and IDF 2003 – All rights reserved 1

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SIST EN ISO 11816-2:2003
ISO 11816-2:2003(E)
IDF 155-2:2003(E)
4 Principle

In the presence of alkaline phosphatase, a non-fluorescent monophosphoric ester substrate (Fluorophos ) is

hydrolysed at 38 °C ± 2 °C for 3 min to form a highly fluorescent product (Fluoroyellow ). The amount of
Fluoroyellow is measured with a calibrated fluorometer and the activity of the alkaline phosphatase is
calculated.
NOTE Although this is a 3-min test, the first minute is an equilibration period to ensure that the sample is at 38 °C.
Measurements of activity are actually made from the beginning of the second minute to the end of the third minute (i.e.
over a 2-min period).
5 Reagents
Use only reagents of recognized analytical grade, unless otherwise specified, and distilled or demineralized
water or water of equivalent purity.
1)
5.1 Fluorophos substrate , crystallized, M = 580 g/mol.
NOTE Fluorophos substrate is a water-soluble, non-fluorescent aromatic monophosphoric ester.
5.2 Substrate buffer solution: diethanolamine (DEA) buffer solution (c = 2,4 mol/l), with pH 10,0.
5.3 Working substrate
Add a volume of the substrate buffer solution (5.2) to the Fluorophos substrate (5.1) to obtain a concentration
of Fluorophos substrate of c = 1,044 mmol/l. Mix well by inversion.
Use amber glass to protect against light.
If using the Fluorophos test system, add the content of 1 bottle of the substrate buffer solution (5.2) to one
bottle of the Fluorophos s
...