EN 14177:2003

Foodstuffs - Determination of patulin in clear and cloudy apple juice and puree - HPLC method with liquid/liquid partition clean-up

This European Standard specifies a method for the determination of patulin in apple juices and apple puree up to 128 ug/kg using high performance liquid chromatography (HPLC).

Lebensmittel - Bestimmung von Patulin in klarem und trübem Apfelsaft und Apfelpüree - HPLC-Verfahren mit Reinigung durch Flüssig/Flüssig-Verteilung

Anwendungsbereich
Dieser Europäische Norm-Entwurf legt ein hochleistungs-flüssigchromatographisches Verfahren (HPLC) zur Bestimmung von Patulin in Apfelsaft und Apfelpüree bis zu einem Massengehalt von 128 µg/kg fest.

Produits alimentaires - Détermination de la teneur en patuline dans le jus de pommes clarifié et trouble et dans la compote de pommes - Méthode par CLHP avec purification par partage liquide/liquide

La présente Norme européenne spécifie une méthode de dosage de la patuline dans le jus de pommes et la compote de pommes par chromatographie en phase liquide à haute performance (CLHP). Cette méthode a été validée pour permettre de doser la patuline en procédant à l'analyse d'échantillons naturellement contaminés et supplémentés de jus de pommes clair à des niveaux compris entre 26 µg/l et 128 µg/l, de jus de pommes trouble à des niveaux compris entre 26 µg/l et 106 µg/l et de compote de pommes à des niveaux compris entre 23 µg/kg et 121 µg/kg.

Živila - Določevanje patulina v bistrem in motnem jabolčnem soku in jabolčni kaši - Metoda s HPLC s čiščenjem s porazdelitvijo tekočina-tekočina

General Information

• Status: Published
• Publication Date: 16-Dec-2003
• ICS: 67.080.10 - Fruits and derived products
• ICS: 67.160.20 - Non-alcoholic beverages
• Technical Committee: CEN/TC 275 - Food analysis - Horizontal methods
• Drafting Committee: CEN/TC 275/WG 5 - Biotoxins
• Current Stage: 9093 - Decision to confirm - Review Enquiry
• Start Date: 09-Dec-2024
• Completion Date: 14-Apr-2025
• Ref Project: SIST EN 14177:2004 - Foodstuffs - Determination of patulin in clear and cloudy apple juice and puree - HPLC method with liquid/liquid partition clean-up

Overview

EN 14177:2003 (CEN) specifies a standardized HPLC method for the determination of patulin in clear and cloudy apple juice and apple puree. The procedure uses an enzyme treatment for cloudy samples, extraction with ethyl acetate, liquid/liquid partition clean-up, drying, solvent evaporation and quantitative determination by HPLC with UV detection. The standard includes validated concentration ranges, precision data and safety warnings for handling patulin.

Key topics and technical requirements

• Scope and validation: Method validated for naturally contaminated and spiked samples in clear juice, cloudy juice and apple puree (validation concentration ranges are provided in the standard).
• Sample preparation:
  • Cloudy juice and puree are treated with an endogalacturonase enzyme to release patulin.
  • Extraction of patulin from 10 ml sample portions using ethyl acetate (three extractions).
  • Liquid/liquid partition clean-up using aqueous sodium carbonate to remove acidic interferences.
  • Drying of organic extract with anhydrous sodium sulfate and evaporation before HPLC analysis.
• Analytical method:
  • HPLC: reverse-phase column (e.g., C18), isocratic mobile phase (water : acetonitrile with perchloric acid additive as specified), UV detection around 276 nm.
  • System details: 50 µl injection loop example, 1 ml/min flow rate, suitable pre-column recommended.
• Reagents and apparatus: analytical-grade solvents (HPLC grade), enzyme solution, patulin stock/standard preparations, centrifuge, rotary evaporator, UV spectrophotometer and HPLC system.
• Safety: patulin is a suspected mutagen and potentially immunotoxic - use gloves, eye protection and perform sample prep in a fume hood.

Practical applications and users

• Food safety laboratories performing routine monitoring of juices and fruit products for mycotoxin compliance.
• Quality control teams in juice manufacturing to verify raw material and finished product safety.
• Regulatory agencies and inspection bodies enforcing maximum levels of patulin in foodstuffs.
• Accreditation and proficiency testing providers using the standardized method for interlaboratory comparisons.

Benefits:

• Provides a harmonized, validated protocol for reliable patulin quantification.
• Supports regulatory compliance, consumer safety and lab accreditation (traceable CEN method).

Related standards

• EN ISO 3696 - “Water for analytical laboratory use” (referenced for reagent water quality).
• Other CEN methods for mycotoxin analysis and foodborne contaminant testing may be relevant for comprehensive monitoring.

Keywords: EN 14177:2003, patulin determination, HPLC method, apple juice testing, apple puree analysis, liquid/liquid partition, CEN standard, food safety testing.

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Bestimmung von Patulin in klarem und trübem Apfelsaft und Apfelpüree - HPLC-Verfahren mit Reinigung durch Flüssig/Flüssig-VerteilungProduits alimentaires - Détermination de la teneur en patuline dans le jus de pommes clarifié et trouble et dans la compote de pommes - Méthode par CLHP avec purification par partage liquide/liquideFoodstuffs - Determination of patulin in clear and cloudy apple juice and puree - HPLC method with liquid/liquid partition clean-up67.080.10Sadje in sadni proizvodiFruits and derived products67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 14177:2003SIST EN 14177:2004en01-maj-2004SIST EN 14177:2004SLOVENSKI
STANDARD
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 14177December 2003ICS 67.080.10; 67.160.20English versionFoodstuffs - Determination of patulin in clear and cloudy applejuice and puree - HPLC method with liquid/liquid partition clean-upProduits alimentaires - Détermination de la teneur enpatuline dans le jus de pommes et dans la compote depommes, limpides et troubles - Méthode parchromatographie en phase liquide à haute performanceavec purification par partage liquide/liquideLebensmittel - Bestimmung von Patulin in klarem undtrübem Apfelsaft und Apfelpüree - HPLC-Verfahren mitReinigung durch Flüssig/Flüssig-VerteilungThis European Standard was approved by CEN on 3 November 2003.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards 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 translationunder the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the officialversions.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 UnitedKingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2003 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 14177:2003 ESIST EN 14177:2004

Precision data.13Bibliography.15SIST EN 14177:2004
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