SIST EN ISO 11816-1:2001

Milk and milk products - Determination of alkaline phosphatase activity using a fluorimetric method - Part 1: Milk and milk-based drinks (ISO 11816-1:1997)

Milch und Milchprodukte - Bestimmung der Aktivität der alkalischen Phosphatase mit einem fluorimetrischen Verfahren - Teil 1: Milch und flüssige Milchprodukte (ISO 11816-1:1997)

Dieser Teil von ISO 11816 legt ein fluorimetrisches Verfahren für die Bestimmung der Aktivität der alkalischen Phosphatase in pasteurisierter Vollmilch, teilentrahmter, entrahmter und aromatisierter Milch fest. Dieses Verfahren ist auch für die Bestimmung der hohen Phosphatase-Aktivität von Rohmilch und von thermisierter Milch mit Aktivitäten von über 7000 Milliunits je Liter geeignet.

Lait et produits laitiers - Détermination de l'activité de la phosphatase alcaline a l'aide de la méthode fluorimétrique - Partie 1: Lait et boissons a base de lait (ISO 11816-1:1997)

IEC 62280:2014 donne les exigences de base nécessaires pour réaliser une communication relative à la sécurité entre des équipements relatifs à la sécurité connectés au système de transmission. La présente norme internationale est applicable à la spécification des exigences de sécurité des équipements relatifs à la sécurité connectés au système de transmission, en vue d'obtenir les exigences d'intégrité de sécurité affectées.

Mleko in mlečni proizvodi - Določevanje aktivnosti alkalne fosfataze s fluorometrijsko metodo - 1. del: Mleko in mlečne pijače (ISO 11816-1:1997)

General Information

• Status: Withdrawn
• Publication Date: 31-Jan-2001
• Withdrawal Date: 31-Aug-2006
• ICS: 67.100.10 - Milk and processed milk products
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 9900 - Withdrawal (Adopted Project)
• Start Date: 01-Sep-2006
• Due Date: 01-Sep-2006
• Completion Date: 01-Sep-2006
• Ref Project: EN ISO 11816-1:2000 - Milk and milk products - Determination of alkaline phosphatase activity using a fluorimetric method - Part 1: Milk and milk-based drinks (ISO 11816-1:1997)

Relations

• Replaced By: SIST EN ISO 22160:2007 - Milk and milk-based drinks - Determination of alkaline phosphatase activity - Enzymatic photo-activated system (EPAS) method (ISO 22160:2007)
• Effective Date: 22-Dec-2008
• Replaced By: SIST EN 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)
• Effective Date: 01-Sep-2006

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN ISO 11816-1:2001
01-februar-2001
0OHNRLQPOHþQLSURL]YRGL'RORþHYDQMHDNWLYQRVWLDONDOQHIRVIDWD]HV
IOXRURPHWULMVNRPHWRGRGHO0OHNRLQPOHþQHSLMDþH,62
Milk and milk products - Determination of alkaline phosphatase activity using a
fluorimetric method - Part 1: Milk and milk-based drinks (ISO 11816-1:1997)
Milch und Milchprodukte - Bestimmung der Aktivität der alkalischen Phosphatase mit
einem fluorimetrischen Verfahren - Teil 1: Milch und flüssige Milchprodukte (ISO 11816-
1:1997)
Lait et produits laitiers - Détermination de l'activité de la phosphatase alcaline a l'aide de
la méthode fluorimétrique - Partie 1: Lait et boissons a base de lait (ISO 11816-1:1997)
Ta slovenski standard je istoveten z: EN ISO 11816-1:2000
ICS:
67.100.10 0OHNRLQSUHGHODQLPOHþQL Milk and processed milk
SURL]YRGL products
SIST EN ISO 11816-1:2001 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 11816-1:2001

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SIST EN ISO 11816-1:2001

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SIST EN ISO 11816-1:2001

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SIST EN ISO 11816-1:2001
INTERNATIONAL IS0
STANDARD 11816-1
First edition
1997-02- 15
Milk and milk products - Determination of
alkaline phosphatase activity using a
fluorimetric method -
Part 1:
Milk and milk-based drinks
Lait et produits laitiers - &termination de I’activit6 de la phosphatase
alcaline 2 /‘aide de la mkthode fluorimktrique -
Partie I: Lait et boissons ZI base de lait
Reference number
IS0 11816-1:1997(E)

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SIST EN ISO 11816-1:2001
IS0 11816-l : 1997(E)
Foreword
IS0 (the International Organization for Standardization) is a worldwide federation of national standards bodies (IS0
member bodies). The work of preparing International Standards is normally carried out through IS0 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. IS0 collaborates closely with the International Electrotechnical
Commission (IEC) on all matters of electrotechnical standardization.
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.
International Standard IS0 11816-I was prepared by Technical Committee lSO/TC 34, Agricultural food products,
Subcommittee SC 5, Milk and milk products, in collaboration with the International Dairy Federation (IDF) and
AOAC INTERNATIONAL, and will also be published by these organizations.
IS0 11816 consists of the following parts, under the general title Milk and milk products - Determination of alkaline
phosphatase activity using a fluorimetric method
- Part I: Milk and milk-based drinks
- Part2: Cheeses
Annex A of this part of IS0 11816 is for information only.
0 IS0 1997
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 the publisher.
International Organization for Standardization
Case postale 56 l CH-1211 Geneve 20 e Switzerland
Internet central @ iso.ch
x.400 c=ch; a=400net; p=iso; o=isocs; s=central
Printed in Switzerland

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SIST EN ISO 11816-1:2001
INTERNATIONAL STANDARD o IS0 IS0 11816-l :1997(E)
Milk and milk products - Determination of alkaline phosphatase
activity using a fluorimetric method -
Part 1:
Milk and milk-based drinks
1 Scope
This part of IS0 11816 specifies a fluorimetric method for the determination of alkaline phosphatase activity in
pasteurized whole milk, semi-skimmed milk, skimmed milk and flavoured milks.
The method is also suitable for the determination of high alkaline phosphatase activity in raw milk and heat-treated
milk having activities of more than 7 000 milliunits per litre.
2 Definitions
For the purposes of this part of IS0 11816, the following definitions apply.
2.1 alkaline phosphatase activity (APL): Activity of the alkaline phosphatase present in a product, determined in
accordance with the procedure specified in this part of IS0 11816. It is expressed as milliunits per litre.
NOTE 1 See reference [I] in annex A.
2.2 unit of alkaline phosphatase activity: Amount of alkaline phosphatase enzyme that catalyses the
transformation of 1 pmol of substrate per minute per litre of sample.
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, 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.
NOTE 2 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).
4 Reagents
Use only reagents of recognized analytical grade, unless otherwise specified, and distilled or demineralized water or
water of equivalent purity.

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SIST EN ISO 11816-1:2001
IS0 11816=1:1997(E) 0 IS0
4.1 Substrate, for example Fluorophos substrate’), crystallized.
NOTE 3 Fluorophos substrate is a water-soluble, non-fluorescent aromatic monophosphoric ester substrate, which is stable
for 1 year when crystallized and stored in glass vials at 4 “C.
4.2 Substrate diluent”: diethanolamine (DEA) buffer (pH lO,O), 2,4 mol/l solution.
NOTE 4 The buffer solution is stable for 1 year at 4 “C.
4.3 Working substrate”
Add a volume of the substrate diluent (4.2) to the substrate (4.1) to give a concentration of 1,044 mmol/l and mix
well by inversion. Use amber glass to protect against light.
If using the Fluorophos Test System”, add the content of 1 vial of the substrate diluent (4.2) to one vial of the
substrate (4.1) and mix well by inversion.
NOTE 5 This solution is stable when stored in the dark for 8 weeks at 4 “C and 8 h at 38 “C. It is sufficient for 115 tests.
4.4 Working calibrators”, for example Fluoroyellow in DEA buffer.
4.4.1 Calibrator solution A, containing 0 pmol/l of Fluoroyellow.
4.4.2 Calibrator solution B, containing 17,24 x IO-3 pmol/l of Fluoroyellow.
4.4.3 Calibrator solution C, containing 34,48 x 10-s kmol/l of Fluoroyellow.
NOTES
6 Calibrator solutions are stable for 1 year when stored at 4 “C.
7 The volumes of reagents supplied for the Fluorophos Test System may be changed by the manufacturer. The user should
refer to the manufacturer’s instructions for preparing the reagents if the volumes supplied are different from those specified in
4.1 to 4.4.
5 Apparatus
Usual laboratory equipment and, in particular, the following.
5.1 Filter fluorimeter”, with thermostatted cuvette holder maintained at 38 “C t 1 “C, and right-angle optics,
allowing excitation at a wavelength of 440 nm and emission at 560 nm.
5.2 Cuvettes, disposable, non-fluorescent glass, of diameter 12 mm and length 75 mm.
5.3 Fixed-volume dispenser, to dispense 2,0 ml.
5.4 Positive-displacement pipette, of capacity 0,075 ml.
5.5 Pipettes, of capacities 1 ml and 2 ml.
‘) The reagents specified in 4.1 to 4.4 and the apparatus specified in 5.1 to 5.6 (except 5.5) are available as the Fluorophos
Test System from Advanced Instruments Inc., Two Technology Way, Norwood, MA 02062, USA. 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 part of IS0 11816 and does not constitute an endorsement by IS0 of
these products.

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SIST EN ISO 11816-1:2001
0 IS0 IS0 11816=1:1997(E)
5.6 Incubator block, suitable to hold the cuvettes and capable of being maintained at a temperature of
38 “C t 1 “C.
5.7 Parafilm or other suitable laboratory grade film.
5.8 Vortex shaker
5.9 Water bath, capable of being maintained at 95 “C t 1 “C.
5.10 One-mark volumetric flasks, of capacity 100 ml.
6 Sampling
which is truly representative and has not been damaged
It is important that the laboratory receive a sample or
changed during transport or storage.
Sampling is not part of the method specified in this part of IS0 11816. A recommended sampling method is given in
IS0 707 [*I.
7 Pr
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