Microbiology of the food chain - Detection and quantification of histamine in fish and fishery products - HPLC method (ISO 19343:2017)

This standard describes the detection and quantification of histamine

Mikrobiologie der Nahrungskette - Nachweis und Bestimmung von Histamin in Fisch und Fischereierzeugnissen - HPLC-Methode (ISO 19343:2017)

Diese Internationale Norm legt eine Hochleistungsflüssigkeitschromatografie-Methode (HPLC) fest, um Histamin in Fisch und Fischereierzeugnissen für die menschliche Ernährung zu titrieren.

Microbiologie de la chaîne alimentaire - Détection et quantification de l'histamine par méthode HPLC (ISO 19343:2017)

L'ISO 19343:2017 spécifie une méthode de chromatographie en phase liquide à haute performance (CLHP) pour analyser l'histamine dans le poisson et les produits de la pêche (sauces au poisson, poissons ayant subi un traitement de maturation enzymatique dans la saumure, etc.) destinés à la consommation humaine.

Mikrobiologija v prehranski verigi - Odkrivanje prisotnosti in kvantifikacija histamina v ribah in ribjih proizvodih - Metoda HPLC (ISO 19343:2017)

Ta standard opisuje način odkrivanja prisotnosti in kvantifikacijo histamina.

General Information

Status
Published
Public Enquiry End Date
19-Jun-2016
Publication Date
15-Aug-2017
Current Stage
6060 - National Implementation/Publication (Adopted Project)
Start Date
01-Aug-2017
Due Date
06-Oct-2017
Completion Date
16-Aug-2017

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SLOVENSKI STANDARD
SIST EN ISO 19343:2017
01-september-2017
Mikrobiologija v prehranski verigi - Odkrivanje prisotnosti in kvantifikacija
histamina v ribah in ribjih proizvodih - Metoda HPLC (ISO 19343:2017)

Microbiology of the food chain - Detection and quantification of histamine in fish and

fishery products - HPLC method (ISO 19343:2017)

Mikrobiologie der Nahrungskette - Nachweis und Bestimmung von Histamin in Fisch und

Fischereierzeugnissen - HPLC-Methode (ISO 19343:2017)

Microbiologie de la chaîne alimentaire - Détection et quantification de l'histamine par

méthode HPLC (ISO 19343:2017)
Ta slovenski standard je istoveten z: EN ISO 19343:2017
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.120.30 Ribe in ribji proizvodi Fish and fishery products
SIST EN ISO 19343:2017 en

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

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SIST EN ISO 19343:2017
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SIST EN ISO 19343:2017
EN ISO 19343
EUROPEAN STANDARD
NORME EUROPÉENNE
July 2017
EUROPÄISCHE NORM
ICS 07.100.30
English Version
Microbiology of the food chain - Detection and
quantification of histamine in fish and fishery products -
HPLC method (ISO 19343:2017)

Microbiologie de la chaîne alimentaire - Détection et Mikrobiologie der Lebensmittelkette - Nachweis und

quantification de l'histamine dans le poisson et les Bestimmung von Histamin in Fisch und

produits de la pêche - Méthode par CLHP (ISO Fischereierzeugnissen - HPLC-Methode (ISO

19343:2017) 19343:2017)
This European Standard was approved by CEN on 8 June 2017.

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, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels

© 2017 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 19343:2017 E

worldwide for CEN national Members.
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SIST EN ISO 19343:2017
EN ISO 19343:2017 (E)
Contents Page

European foreword ....................................................................................................................................................... 3

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SIST EN ISO 19343:2017
EN ISO 19343:2017 (E)
European foreword

This document (EN ISO 19343:2017) has been prepared by Technical Committee ISO/TC 34 “Food

products” in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”

the secretariat of which is held by DIN.

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 January 2018 and conflicting national standards shall

be withdrawn at the latest by January 2018.

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. CEN shall not be held responsible for identifying any or all such patent rights.

This document has been prepared under a mandate given to CEN by the European Commission and the

European Free Trade Association.

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, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland,

Turkey and the United Kingdom.
Endorsement notice

The text of ISO 19343:2017 has been approved by CEN as EN ISO 19343:2017 without any modification.

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SIST EN ISO 19343:2017
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SIST EN ISO 19343:2017
INTERNATIONAL ISO
STANDARD 19343
First edition
2017-06
Microbiology of the food chain —
Detection and quantification of
histamine in fish and fishery products
— HPLC method
Microbiologie de la chaîne alimentaire — Détection et quantification de
l’histamine dans le poisson et les produits de la pêche — Méthode CLHP
Reference number
ISO 19343:2017(E)
ISO 2017
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SIST EN ISO 19343:2017
ISO 19343:2017(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2017, Published in Switzerland

All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form

or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior

written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of

the requester.
ISO copyright office
Ch. de Blandonnet 8 • CP 401
CH-1214 Vernier, Geneva, Switzerland
Tel. +41 22 749 01 11
Fax +41 22 749 09 47
copyright@iso.org
www.iso.org
ii © ISO 2017 – All rights reserved
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SIST EN ISO 19343:2017
ISO 19343:2017(E)
Contents Page

Foreword ........................................................................................................................................................................................................................................iv

Introduction ..................................................................................................................................................................................................................................v

1 Scope ................................................................................................................................................................................................................................. 1

2 Normative references ...................................................................................................................................................................................... 1

3 Terms and definitions ..................................................................................................................................................................................... 1

4 Principle ........................................................................................................................................................................................................................ 1

5 Reagents and materials ................................................................................................................................................................................. 1

6 Apparatus ..................................................................................................................................................................................................................... 2

7 Procedure..................................................................................................................................................................................................................... 3

7.1 Sample preparation ............................................................................................................................................................................ 3

7.2 Extraction .................................................................................................................................................................................................... 3

7.3 Derivatization ........................................................................................................................................................................................... 3

7.4 Purification ................................................................................................................................................................................................. 3

7.5 LC condition ............................................................................................................................................................................................... 4

7.6 Range of standard sample ............................................................................................................................................................. 4

8 Calculation .................................................................................................................................................................................................................. 4

8.1 Calibration line (curve) .................................................................................................................................................................... 4

8.2 Histamine quantification ................................................................................................................................................................ 5

9 Precision ....................................................................................................................................................................................................................... 5

9.1 Interlaboratory study ........................................................................................................................................................................ 5

9.2 Repeatability limit ................................................................................................................................................................................ 5

9.3 Reproducibility limit .......................................................................................................................................................................... 5

Annex A (informative) Recommendations for HPLC separation ............................................................................................. 7

Annex B (informative) Performance characteristics of the method .................................................................................... 9

Bibliography .............................................................................................................................................................................................................................14

© ISO 2017 – All rights reserved iii
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SIST EN ISO 19343:2017
ISO 19343:2017(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.

The procedures used to develop this document and those intended for its further maintenance are

described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the

different types of ISO documents should be noted. This document was drafted in accordance with the

editorial rules of the ISO/IEC Directives, Part 2 (see www .iso .org/ directives).

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. Details of

any patent rights identified during the development of the document will be in the Introduction and/or

on the ISO list of patent declarations received (see www .iso .org/ patents).

Any trade name used in this document is information given for the convenience of users and does not

constitute an endorsement.

For an explanation on the voluntary nature of standards, the meaning of ISO specific terms and

expressions related to conformity assessment, as well as information about ISO’s adherence to the

World Trade Organization (WTO) principles in the Technical Barriers to Trade (TBT) see the following

URL: w w w . i s o .org/ iso/ foreword .html.

This document was prepared by the European Committee for Standardization (CEN) Technical

Committee CEN/TC 275, Food analysis — Horizontal methods, in collaboration with ISO Technical

Committee TC 34, Food products, Subcommittee SC 9, Microbiology, in accordance with the agreement

on technical cooperation between ISO and CEN (Vienna Agreement).
iv © ISO 2017 – All rights reserved
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SIST EN ISO 19343:2017
ISO 19343:2017(E)
Introduction

Histamine is a causative agent of scombroid poisoning or histamine fish poisoning. Histamine can

be present mainly in Scombridae (tuna, mackerel) and Clupeidae (herring, sardine), species which

contain a high level of free histidine. Histamine is formed through the decarboxylation of histidine by

microbiological histidine decarboxylase.

Histamine [2-(1H-imidazol-5-yl)ethanamine] is defined as a biologically active low molecular weight

basic nitrogenous molecule. The consumption of food containing significant concentration of histamine

can cause symptoms similar to those associated to food allergies.

This document was developed in response to the need to standardize a method for histamine detection

[1]

and quantification in fish and fishery products, in particular for European Regulation 2073/2005 on

microbiological criteria for foodstuffs.
© ISO 2017 – All rights reserved v
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SIST EN ISO 19343:2017
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SIST EN ISO 19343:2017
INTERNATIONAL STANDARD ISO 19343:2017(E)
Microbiology of the food chain — Detection and
quantification of histamine in fish and fishery products —
HPLC method

WARNING — Persons using this document should be familiar with normal laboratory practice.

This document does not purport to address all of the safety problems, if any, associated with its

use. It is the responsibility of the user to establish appropriate safety and health practices.

1 Scope

This document specifies a high performance liquid chromatography (HPLC) method to analyse

histamine in fish and fishery products (fish sauces, fish maturated by enzyme in brine, etc.) intended

for human consumption.
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements 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 3696, Water for analytical laboratory use — Specification and test methods
3 Terms and definitions
No terms and definitions are listed in this document.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— IEC Electropedia: available at http:// www .electropedia .org/
— ISO Online browsing platform: available at http:// www .iso .org/ obp
4 Principle

This method enables the separation of histamine among biogenic amines in fish and fishery products.

The sample is extracted by mixing with perchloric acid. Precolumn derivatization is performed using

dansyl chloride. The biogenic amines and the components in the solution are separated by HPLC with

an appropriate column, using UV detection. Histamine mass concentration is calculated from the peak

area ratio of histamine and internal standard with a calibration curve.
5 Reagents and materials

Use only reagents of recognized analytical grade and water complying with grade 1 of ISO 3696, unless

otherwise specified. Solvents shall be of quality for HPLC analysis, unless otherwise specified.

5.1 Acetone, CH COCH .
3 3
5.2 Acetonitrile, CH CN.
5.3 Toluene, C H .
7 8
© ISO 2017 – All rights reserved 1
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SIST EN ISO 19343:2017
ISO 19343:2017(E)
5.4 Water, HPLC grade.
5.5 Water, distilled or equivalent.
5.6 Nitrogen, gas.
5.7 Perchloric acid, c(HClO ) = 0,2 mol/l (recommended).

Dilute 19,5 ml of HClO 65 % or 17,2 ml of HClO 70 % to 1 000 ml of water (5.5). The solution is stable

4 4
for six months if stored at room temperature (15°C to 25°C).
5.8 Saturated sodium carbonate solution, Na CO .
2 3

Dissolve 110 g of sodium carbonate in about 150 ml of water (5.5). The solution is in excess and stable

for three months if stored at 5 °C ± 3°C.
5.9 Dansyl chloride solution, ρ(C H ClNO S) = 7,5 mg/ml.
12 12 2

Dissolve 0,375 g of dansyl chloride in 50 ml of acetone (5.1). The solution is stable for three weeks if

stored in the dark at a temperature lower than < −18 °C.
5.10 L-proline solution, ρ(C H NO ) = 100 mg
...

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