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

2

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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.
3

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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

---------------------- Page: 8 ----------------------

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.
3
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).
4
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
...

SLOVENSKI STANDARD
oSIST prEN ISO 19343:2016
01-junij-2016
Mikrobiologija v prehranski verigi - Odkrivanje prisotnosti in kvantifikacija
histamina v ribah in ribjih proizvodih - Metoda HPLC (ISO/DIS 19343:2016)
Microbiology of the food chain - Detection and quantification of histamine in fish and
fishery products - HPLC method (ISO/DIS 19343:2016)
Mikrobiologie der Nahrungskette - Nachweis und Bestimmung von Histamin in Fisch und
Fischereierzeugnissen - HPLC-Methode (ISO/DIS 19343:2016)
Microbiologie de la chaîne alimentaire - Détection et quantification de l'histamine par
méthode HPLC (ISO/DIS 19343:2016)
Ta slovenski standard je istoveten z: prEN ISO 19343
ICS:
07.100.30 Mikrobiologija živil Food microbiology
67.120.30 Ribe in ribji proizvodi Fish and fishery products
oSIST prEN ISO 19343:2016 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
oSIST prEN ISO 19343:2016

---------------------- Page: 2 ----------------------
oSIST prEN ISO 19343:2016
DRAFT INTERNATIONAL STANDARD
ISO/DIS 19343
ISO/TC 34/SC 9 Secretariat: AFNOR
Voting begins on: Voting terminates on:
2016-03-31 2016-06-29
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 par méthode HPLC
ICS: 07.100.30
ISO/CEN PARALLEL PROCESSING
This draft has been developed within the European Committee for Standardization
(CEN), and processed under the CEN lead mode of collaboration as defined in the
Vienna Agreement.
This draft is hereby submitted to the ISO member bodies and to the CEN member
bodies for a parallel three month enquiry.
THIS DOCUMENT IS A DRAFT CIRCULATED
FOR COMMENT AND APPROVAL. IT IS
THEREFORE SUBJECT TO CHANGE AND MAY
NOT BE REFERRED TO AS AN INTERNATIONAL
STANDARD UNTIL PUBLISHED AS SUCH.
To expedite distribution, this document is circulated as received from the
IN ADDITION TO THEIR EVALUATION AS
committee secretariat. ISO Central Secretariat work of editing and text
BEING ACCEPTABLE FOR INDUSTRIAL,
composition will be undertaken at publication stage.
TECHNOLOGICAL, COMMERCIAL AND
USER PURPOSES, DRAFT INTERNATIONAL
STANDARDS MAY ON OCCASION HAVE TO
BE CONSIDERED IN THE LIGHT OF THEIR
POTENTIAL TO BECOME STANDARDS TO
WHICH REFERENCE MAY BE MADE IN
Reference number
NATIONAL REGULATIONS.
ISO/DIS 19343:2016(E)
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. ISO 2016

---------------------- Page: 3 ----------------------
oSIST prEN ISO 19343:2016
ISO/DIS 19343:2016(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2016, 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 2016 – All rights reserved

---------------------- Page: 4 ----------------------
oSIST prEN ISO 19343:2016
ISO/DIS 19343:2016(E)
Contents
Foreword.4
Introduction .5
1 Scope .1
2 Principle .1
3 Reagents and products .1
4 Apparatus .2
5 Procedure .3
5.1 Sample preparation .3
5.2 Extraction .3
5.3 Derivatization .3
5.4 Purification .3
5.5 LC conditions .4
5.6 Standard samples.4
6 Calculation.4
6.1 Calibration line (curve) .4
6.2 Histamine quantification.4
Annex A (informative) Recommendations for HPLC separation.6
Annex B (informative) Precision and trueness .8
Bibliography . 11

© ISO 2016 – All rights reserved iii

---------------------- Page: 5 ----------------------
oSIST prEN ISO 19343:2016
ISO/DIS 19343:2016(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 meaning of ISO specific terms and expressions related to conformity
assessment, as well as information about ISO’s adherence to the WTO principles in the Technical
Barriers to Trade (TBT) see the following URL: Foreword - Supplementary information.
ISO 19343 was prepared the European Committee for Standardization (CEN) in collaboration with
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 2016 – All rights reserved

---------------------- Page: 6 ----------------------
oSIST prEN ISO 19343:2016
ISO/DIS 19343:2016(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), in species which
contain a high level of free histidine. Histamine is formed through the decarboxylation of histidine by
microbiological histidine decarboxylase.
Histamine is defined as a biologically active low molecular weight basic nitrogenous molecule. Th e
consumption of food containing significant concentration of histamine can cause symptoms similar to
those associated to seafood allergies.
This standard was developed in response to the need of standardizing a method for histamine detection
and quantification in fish and fishery products, in particular for the European regulation 2073/2005 on
microbiogical criteria for foodstuffs.
© ISO 2016 – All rights reserved v

---------------------- Page: 7 ----------------------
oSIST prEN ISO 19343:2016

---------------------- Page: 8 ----------------------
oSIST prEN ISO 19343:2016
ISO/DIS 19343:2016(E)
Microbiology of the food chain — Detection and quantification of
histamine in fish and fishery products — HPLC method
1 Scope
This International Standard specifies a High Performance Liquid Chromatography (HPLC) method to
titrate histamine in fish and fishery products intended for human consumption.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated
references, the latest edition of the referenced document (including any amendments) applies.
EN ISO 3696, Water for analytical laboratory use — Specification and test methods (ISO 3696)
3 Principle
This method enables to separate histamine among biogenic amines of 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
appropriate column, using UV detection. After derivatization, histamine mass concentration is
calculated from the peak area ratio of histamine and internal standard, with a calibration curve.
4 Reagents and materials
Use only reagents of recognized analytical grade and water complying with grade 1 of EN ISO 3696,
unless otherwise specified. Solvents shall be of quality for HPLC analysis, unless otherwise specified.
4.1 Acetone, CH3COCH3.
4.2 Acetonitrile, HPLC quality, CH CN.
3
4.3 Toluene, C H .
7 8
4.4 Water HPLC quality.
4.5 Bidistilled water.
4.6 Nitrogen.
4.7 Perchloric acid, substance concentration c (HClO ) = 0,2 mol/l (recommended).
4
Add 19,5 ml of HClO 65 % or 17,2 ml of HClO 70 % to 1 000 ml of water (3.4). The solution is stable for
4 4
six months if stored at room temperature (15°C to 25°C).
© ISO 2016 – All rights reserved 1

---------------------- Page: 9 ----------------------
oSIST prEN ISO 19343:2016
ISO/DIS 19343:2016(E)
4.8 Sodium carbonate, (Na CO ) solution.
2 3
Dissolve 110 g of sodium carbonate until complete saturation in about 150 ml of water (3.4). The
soluti
...

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