SIST EN ISO 11816-1:2006
(Main)Milk and milk products - Determination of alkaline phosphatase activity - Part 1: Fluorimetric method for milk and milk-based drinks (ISO 11816-1:2006)
Milk and milk products - Determination of alkaline phosphatase activity - Part 1: Fluorimetric method for milk and milk-based drinks (ISO 11816-1:2006)
This part of ISO 11816 IDF 155 specifies a fluorimetric method for the determination of alkaline phosphatase (ALP, EC 3.1.3.1) activity in pasteurized whole milk, semi-skimmed milk, skimmed milk and flavoured milks. The method is applicable for milk from cows, sheep and goats, and milk-based drinks. The method is also suitable for the determination of high alkaline phosphatase activity in raw milk and heattreated milk with activities of more than 2 000 mU/l after dilution of the sample as specified.
Milch und Milchprodukte - Bestimmung der Aktivität der alkalischen Phosphatase - Teil 1: Fluorimetrisches Verfahren für Milch und flüssige Milchprodukte (ISO 11816-1:2006)
Dieser Teil von ISO 11816 | IDF 155 legt ein fluorimetrisches Verfahren für die Bestimmung der Aktivität der alkalischen Phosphatase (ALP, EC 3.1.3.1) in pasteurisierter Vollmilch, fettarmer Milch, Magermilch und aromatisierter Milch fest. Das Verfahren ist für Milch von Kühen, Schafen und Ziegen und flüssige Milchprodukte anwendbar.
Dieses Verfahren ist auch für die Bestimmung der hohen Phosphatase-Aktivität von Rohmilch und von hitzebehandelter Milch mit Aktivitäten von über 2 000 Milliunits je Liter (mU/l) nach Verdünnung der Probe geeignet.
Lait et produits laitiers - Détermination de l'activité de la phosphatase alcaline - Partie 1: Méthode fluorimétrique pour le lait et les boissons a base de lait (ISO 11816-1:2006)
L'ISO 11816-1|FIL 155-1:2006 décrit une méthode fluorimétrique pour la détermination de l'activité de la phosphatase alcaline (ALP, EC 3.1.3.1) dans les laits pasteurisés entiers, demi-écrémés, écrémés et aromatisés. La présente méthode est applicable aux laits de vache, de brebis et de chèvre, et aux boissons à base de lait.
La méthode convient également pour la détermination d'activités élevées de la phosphatase alcaline dans le lait cru et le lait traité thermiquement, ayant des activités de plus de 2 000 milliunités par litre (mU/l) après dilution de l'échantillon, comme spécifié.
Mleko in mlečni proizvodi - Določevanje aktivnosti alkalne fosfataze s fluorometrijsko metodo - 1. del: Fluorometrična metoda za mleko in mlečne pijače (ISO 11816-1:2006)
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST EN ISO 11816-1:2006
01-september-2006
1DGRPHãþD
SIST EN ISO 11816-1:2001
0OHNRLQPOHþQLSURL]YRGL'RORþHYDQMHDNWLYQRVWLDONDOQHIRVIDWD]HV
IOXRURPHWULMVNRPHWRGRGHO )OXRURPHWULþQDPHWRGD]DPOHNRLQPOHþQHSLMDþH
,62
Milk and milk products - Determination of alkaline phosphatase activity - Part 1:
Fluorimetric method for milk and milk-based drinks (ISO 11816-1:2006)
Milch und Milchprodukte - Bestimmung der Aktivität der alkalischen Phosphatase - Teil 1:
Fluorimetrisches Verfahren für Milch und flüssige Milchprodukte (ISO 11816-1:2006)
Lait et produits laitiers - Détermination de l'activité de la phosphatase alcaline - Partie 1:
Méthode fluorimétrique pour le lait et les boissons a base de lait (ISO 11816-1:2006)
Ta slovenski standard je istoveten z: EN ISO 11816-1:2006
ICS:
67.100.10 0OHNRLQSUHGHODQLPOHþQL Milk and processed milk
SURL]YRGL products
SIST EN ISO 11816-1:2006 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST EN ISO 11816-1:2006
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SIST EN ISO 11816-1:2006
EUROPEAN STANDARD
EN ISO 11816-1
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2006
ICS 67.100.10
English Version
Milk and milk products - Determination of alkaline phosphatase
activity - Part 1: Fluorimetric method for milk and milk-based
drinks (ISO 11816-1:2006)
Lait et produits laitiers - Détermination de l'activité de la Milch und Milchprodukte - Bestimmung der Aktivität der
phosphatase alcaline - Partie 1: Méthode fluorimétrique alkalischen Phosphatase - Teil 1: Fluorimetrisches
pour le lait et les boissons à base de lait (ISO 11816- Verfahren für Milch und flüssige Milchprodukte (ISO 11816-
1:2006) 1:2006)
This European Standard was approved by CEN on 27 March 2006.
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 Central Secretariat 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 Central Secretariat has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,
Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania,
Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: rue de Stassart, 36 B-1050 Brussels
© 2006 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 11816-1:2006: E
worldwide for CEN national Members.
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SIST EN ISO 11816-1:2006
EN ISO 11816-1:2006 (E)
Foreword
This document (EN ISO 11816-1:2006) 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 October 2006, and conflicting national
standards shall be withdrawn at the latest by October 2006.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of
the following countries are bound to implement this European Standard: Austria, Belgium,
Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary,
Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
Endorsement notice
The text of ISO 11816-1:2006 has been approved by CEN as EN ISO 11816-1:2006 without any
modifications.
2
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SIST EN ISO 11816-1:2006
INTERNATIONAL ISO
STANDARD 11816-1
IDF
155-1
Second edition
2006-04-15
Milk and milk products — Determination
of alkaline phosphatase activity —
Part 1:
Fluorimetric method for milk
and milk-based drinks
Lait et produits laitiers — Détermination de l'activité de la phosphatase
alcaline
Partie 1: Méthode fluorimétrique pour le lait et les boissons à base de
lait
Reference numbers
ISO 11816-1:2006(E)
IDF 155-1:2006(E)
©
ISO and IDF 2006
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SIST EN ISO 11816-1:2006
ISO 11816-1:2006(E)
IDF 155-1:2006(E)
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All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
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Published in Switzerland
ii © ISO and IDF 2006 – All rights reserved
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SIST EN ISO 11816-1:2006
ISO 11816-1:2006(E)
IDF 155-1:2006(E)
Contents Page
Foreword. iv
1 Scope . 1
2 Terms and definitions. 1
3 Principle. 1
4 Reagents. 2
5 Apparatus . 3
6 Sampling. 3
7 Preparations . 3
7.1 Alkaline phosphatase-free milk. 3
7.2 Preparation of test sample. 4
8 Procedure . 4
8.1 Verification of instrument performance . 4
8.2 Calibration . 4
8.3 Determination. 5
8.4 Control tests. 5
9 Calculation and expression of results. 7
9.1 Calibration ratio . 7
9.2 Calculation. 7
9.3 Expression of test results. 7
10 Precision. 8
10.1 Interlaboratory test . 8
10.2 Repeatability. 8
10.3 Reproducibility. 8
11 Test report . 8
Annex A (informative) Interlaboratory trial . 9
Bibliography . 11
© ISO and IDF 2006 – All rights reserved iii
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SIST EN ISO 11816-1:2006
ISO 11816-1:2006(E)
IDF 155-1:2006(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 11816-1⎪IDF 155-1 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee
SC 5, Milk and milk products, and the International Dairy Federation (IDF). It is being published jointly by ISO
and IDF.
This edition of ISO 11816-1⎪IDF 155-1 cancels and replaces ISO 11816-1:1997, which has been technically
revised.
ISO 11816⎪IDF 155 consists of the following parts, under the general title Milk and milk products —
Determination of alkaline phosphatase activity:
⎯ Part 1: Fluorimetric method for milk and milk-based drinks
⎯ Part 2: Fluorimetric method for cheese
iv © ISO and IDF 2006 – All rights reserved
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SIST EN ISO 11816-1:2006
ISO 11816-1:2006(E)
IDF 155-1:2006(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
the 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 11816-1⎪IDF 155-1 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 the Joint ISO-IDF Action Team Characterization to heat treatment, of the Standing
Committee on Minor components and characterization of physical properties, under the aegis of its project
leader, Mr F. Harding (UK).
This edition of ISO 11816-1⎪IDF 155-1 cancels and replaces IDF 155A:1999, which has been technically
revised.
ISO 11816⎪IDF 155 consists of the following parts, under the general title Milk and milk products —
Determination of alkaline phosphatase activity:
⎯ Part 1: Fluorimetric method for milk and milk-based drinks
⎯ Part 2: Fluorimetric method for cheese
© ISO and IDF 2006 – All rights reserved v
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SIST EN ISO 11816-1:2006
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SIST EN ISO 11816-1:2006
ISO 11816-1:2006(E)
INTERNATIONAL STANDARD
IDF 155-1:2006(E)
Milk and milk products — Determination of alkaline
phosphatase activity —
Part 1:
Fluorimetric method for milk and milk-based drinks
1 Scope
This part of ISO 11816⎪IDF 155 specifies a fluorimetric method for the determination of alkaline phosphatase
(ALP, EC 3.1.3.1) activity in pasteurized whole milk, semi-skimmed milk, skimmed milk and flavoured milks.
The method is applicable for milk from cows, sheep and goats, and milk-based drinks.
The method is also suitable for the determination of high alkaline phosphatase activity in raw milk and heat-
treated milk with activities of more than 2 000 mU/l after dilution of the sample as specified.
2 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
2.1
alkaline phosphatase activity
APL activity
activity of the alkaline phosphatase present in the product, determined by the procedure specified in this part
of ISO 11816⎪IDF 155
NOTE The alkaline phosphatase activity is expressed as milliunits of enzyme activity per litre (mU/l).
2.2
unit of alkaline phosphatase activity
amount of alkaline phosphatase enzyme that catalyses the transformation of 1 µmol of substrate per minute
3 Principle
The alkaline phosphatase activity of the sample is measured by a continuous fluorimetric direct kinetic assay.
A non-fluorescent aromatic monophosphoric ester substrate, 2′-[2-benzothiazolyl]-6′-hydroxybenzothiazole
phosphate, in the presence of any alkaline phosphatase derived from the sample, undergoes hydrolysis of its
phosphate radical, producing a highly fluorescent product. Fluorimetric measurement of alkaline phosphatase
(ALP) activity is measured at 38 °C over a 3-min period when using the substrate. This includes pre-
incubation of substrate and sample, followed by multiple kinetic readings of the reaction rate.
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).
© ISO and IDF 2006 – All rights reserved 1
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SIST EN ISO 11816-1:2006
ISO 11816-1:2006(E)
IDF 155-1:2006(E)
4 Reagents
Use only reagents of recognized analytical grade, unless otherwise specified, and distilled or demineralized
water or water of equivalent purity.
® 1) ®
4.1 Fluorophos substrate , in bottles, each containing 144 mg of Fluorophos substrate powder.
This is a non-fluorescent aromatic monophosphoric ester substrate, 2′-[2-benzothiazolyl]-6′-hydroxybenzothiazole
® ®
phosphate (Fluorophos ). The Fluorophos substrate remains stable for 2 years when stored in unopened
bottles at between 2 °C and 8 °C.
4.2 Substrate buffer solution, diethanolamine (DEA) buffer solution, c(DEA) = 2,4 mol/l, with pH 10,0, in
bottles of 240 ml each.
The substrate buffer solution remains stable for 2 years when stored in unopened bottles at between 2 °C and
8 °C.
4.3 Working substrate
®
Allow the Fluorophos substrate (4.1) and the substrate buffer solution (4.2) to come to room temperature.
®
Add the content of one bottle of substrate buffer solution (240 ml) (4.2) to that of one bottle Fluorophos
substrate (144 mg) (4.1) and mix well by inversion for 3 min. Use amber glass to protect against light.
Allow the obtained solution stand at room temperature for at least 30 min prior to use.
Use the analog-to-digital test given in 8.4.1.1 to test the suitability of the ready to use working substrate.
The working substrate remains stable for 60 days when protected from light and stored at between 2 °C and
8 °C, or for 8 h when stored at 38 °C. Do not use the working substrate if a reading above 1 200 is obtained
(see 8.4.1.1.5).
NOTE The obtained volume of the working substrate (240 ml) is sufficient for approximately 115 tests.
®
4.4 Working calibrator solutions, Fluoroyellow (FY) [2′-(2-benzothiazolyl)-6′-hydroxybenzothiazole] in
DEA buffer (4.2).
The working calibrator solutions remain stable for 18 months when stored at between 2 °C and 8 °C.
®
4.4.1 Calibrator solution A, containing 0 µmol/l of Fluoroyellow .
–3 ®
4.4.2 Calibrator solution B, containing 17,24 × 10 µmol/l of Fluoroyellow .
–3 ®
4.4.3 Calibrator solution C, containing 34,48 × 10 µmol/l of Fluoroyellow .
–3 ®
4.5 Daily instrument control solution, containing 34,48 × 10 µmol/l of Fluoroyellow .
1) The reagents specified in 4.1 to 4.5 and the apparatus specified in 5.1 to 5.4 (except 5.3.3) are available as
Fluorophos Test System from Advanced Instruments, Inc., Two Technology Way, Norwood, Massachusetts 02062, USA.
The manufacturer may change packaging configurations supplied with the Fluorophos Test System. The user should refer
to the manufacturer’s instructions for preparing reagents if different from those specified herein. Fluorophos and
Fluoroyellow are registered trademarks of Advanced Instruments, Inc. and are examples of suitable products available
commercially.
This information is given for the convenience of users of this document and does not constitute an endorsement by either
ISO or IDF of these products.
2 © ISO and IDF 2006 – All rights reserved
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