EN 15850:2010

Foodstuffs - Determination of zearalenone in maize based baby food, barley flour, maize flour, polenta, wheat flour and cereal based foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection

This European Standard specifies a method for the determination of zearalenone in maize based baby food, barley flour, maize flour, polenta, wheat flour and cereal based foods for infants and young children by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection. This method has been validated in two interlaboratory studies. The first study was for the analysis of samples of maize based baby food, barley flour, maize flour, polenta and wheat flour ranging from 10 µg/kg to 335 µg/kg, and the second study was for samples of cereal based foods for infants and young children ranging from 9 µg/kg to 44 µg/kg.

Lebensmittel - Bestimmung von Zearalenon in Säuglingsnahrung auf Maisbasis, Gerstenmehl, Maismehl, Maisgrieß, Weizenmehl und Lebensmittel auf Getreidebasis für Säuglinge und Kleinkinder - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion

Diese Europäische Norm legt ein Verfahren zur Bestimmung von Zearalenon in Säuglingsnahrung auf Maisbasis, Gerstenmehl, Maismehl, Maisgrieß, Weizenmehl und Lebensmittel auf Getreidebasis für Säuglinge und Kleinkinder durch Hochleistungsflüssigchromatographie (HPLC) mit Reinigung an einer Immunoaffinitäts-säule und Fluoreszenzdetektion fest. Dieses Verfahren wurde in zwei Ringversuchen validiert. Der erste Ringversuch wurde für die Analyse von Säuglingsnahrung auf Maisbasis, Gerstenmehl, Maismehl, Maisgrieß und Weizenmehl mit Gehalten von 10 µg/kg bis 335 µg/kg und der zweite Ringversuch für Säuglings- und Kleinkindernahrung auf Getreidebasis mit Gehalten von 9 µg/kg bis 44 µg/kg durchgeführt.
Weitere Informationen zur Validierung befinden sich in Abschnitt 9 und Anhang B.

Produits alimentaires - Dosage de la zéaralénone dans les aliments à base de maïs pour bébés, dans la farine d'orge, dans la farine de maïs, dans la polenta, dans la farine de blé et dans les aliments à base de céréales pour nourrissons et enfants en bas âge - Méthode par chromatographie liquide haute performance avec purification sur colonne d'immuno-affinité et détection par fluorescence

La présente Norme européenne spécifie une méthode pour déterminer la teneur en zéaralénone dans les aliments
à base de maïs pour bébés, dans la farine d’orge, dans la farine de maïs, dans la polenta, dans la farine de blé et
dans les aliments à base de céréales pour nourrissons et enfants en bas âge, par chromatographie liquide haute
performance (CLHP) avec purification sur colonne d’immuno-affinité et détection fluorimétrique. La présente
méthode a été validée dans deux études interlaboratoires, la première portant sur l’analyse d’échantillons
d’aliments à base de maïs pour bébés, de farine d’orge, de farine de maïs, de polenta et de farine de blé, à des
concentrations allant de 10 μg/kg à 335 μg/kg et la seconde sur l’analyse d’échantillons d’aliments à base de
céréales pour nourrissons et enfants en bas âge à des concentrations allant de 9 μg/kg à 44 μg/kg.
Pour plus d’informations sur la validation, voir l’Article 9 et l’Annexe B.

Živila - Določevanje zearalenona v hrani za dojenčke in majhne otroke na osnovi koruze, ječmenove moke, koruzne moke, polente, pšenične moke ter v žitnih kašicah - Metoda HPLC z imunoafinitetnim kolonskim čiščenjem in fluorescenčno detekcijo

Ta evropski standard določa metodo za določevanje zearalenona v hrani za dojenčke na osnovi koruze, ječmenove moke, koruzne moke, polente, pšenične moke in žitnih kašicah za dojenčke in majhne otroke z tekočinsko kromatografijo visoke ločljivosti (HPLC) z imunoafinitetnim čiščenjem in fluorescenčno detekcijo. Ta metoda je bila potrjena v dveh medlaboratorijskih raziskavah. Prva raziskava je bila izvedena za analizo vzorcev hrane za dojenčke na osnovi koruze, ječmenove moke, koruzne moke, polente in pšenične moke v razponu od 10 µg/kg do 335 µg/kg, druga raziskava pa je bila za vzorce hrane za dojenčke in majhne otroke na osnovi žit v razponu od 9 µg/kg do 44 µg/kg. Za dodatne informacije, glej Klavzulo 9 in Dodatek B.

General Information

• Status: Published
• Publication Date: 06-Apr-2010
• Withdrawal Date: 30-Oct-2010
• ICS: 67.060 - Cereals, pulses and derived products
• 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
• Completion Date: 28-Oct-2020
• Mandate: M/383 - Mandate for standardisation addressed to CEN in the field of Methods of analysis for Mycotoxins in food
• Ref Project: SIST EN 15850:2010 - Foodstuffs - Determination of zearalenone in maize based baby food, barley flour, maize flour, polenta, wheat flour and cereal based foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.OXRUHVFHQþQRGHWHNFLMRLebensmittel - Bestimmung von Zearalenon in Säuglingsnahrung auf Maisbasis, Gerstenmehl, Maismehl, Maisgrieß, Weizenmehl und Lebensmittel auf Getreidebasis für Säuglinge und Kleinkinder - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und FluoreszenzdetektionProduits alimentaires - Dosage de la zéaralénone dans les aliments à base de maïs pour bébés, dans la farine d'orge, dans la farine de maïs, dans la polenta, dans la farine de blé et dans les aliments à base de céréales pour nourrissons et enfants en bas âge - Méthode par chromatographie liquide haute performance avec purification sur colonne d'immuno-affinité et détection par fluorescenceFoodstuffs - Determination of zearalenone in maize based baby food, barley flour, maize flour, polenta, wheat flour and cereal based foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection67.230Predpakirana in pripravljena hranaPrepackaged and prepared foods67.060QMLKCereals, pulses and derived productsICS:Ta slovenski standard je istoveten z:EN 15850:2010SIST EN 15850:2010en,fr,de01-september-2010SIST EN 15850:2010SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 15850
April 2010 ICS 67.060 English Version
Foodstuffs - Determination of zearalenone in maize based baby food, barley flour, maize flour, polenta, wheat flour and cereal based foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection
Produits alimentaires - Dosage de la zéaralénone dans la farine d'orge, de maïs et de blé, la polenta et les produits pour nourrissons et jeunes enfants à base de céréales - Méthode par chromatographie liquide haute performance avec purification sur colonne d'immunoaffinité et détection par fluorescence
Lebensmittel - Bestimmung von Zearalenon in Säuglingsnahrung auf Maisbasis, Gerstenmehl, Maismehl, Maisgrieß, Weizenmehl und Lebensmittel auf Getreidebasis für Säuglinge und Kleinkinder - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion This European Standard was approved by CEN on 27 February 2010.
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 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 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, 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:
Avenue Marnix 17,
B-1000 Brussels © 2010 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 15850:2010: ESIST EN 15850:2010

Typical chromatograms . 15Annex B (informative)
Precision data . 16Bibliography . 18 SIST EN 15850:2010

Specification and test methods (ISO 3696:1987). 3 Principle A test portion is extracted with aqueous acetonitrile or methanol according to the products analyzed. The extract is then diluted with phosphate buffered saline (PBS) to give an aqueous extract that is applied to an immunoaffinity column containing antibodies specific for zearalenone. Zearalenone is purified and concentrated on the column and removed from the antibodies using acetonitrile or methanol as eluent. Zearalenone is quantified by reverse-phase high performance liquid chromatography (RP-HPLC) with fluorescence detection. 4 Reagents 4.1 General Use only reagents of recognised analytical grade and water complying with grade 1 of EN ISO 3696:1995, unless otherwise specified. Solvents shall be of quality for HPLC analysis, unless otherwise specified. Commercially available solutions with equivalent properties to those listed may be used. 4.2 Disodium hydrogen phosphate, Na2HPO4 anhydrous or Na2HPO4·12 H2O. 4.3 Potassium chloride (KCl). 4.4 Potassium dihydrogen phosphate, KH2PO4. 4.5 Sodium chloride (NaCl). 4.6 Sodium hydroxide (NaOH). 4.7 Hydrochloric acid solution, mass fraction w(HCl) = 37 % in water. 4.8 Hydrochloric acid solution, substance concentration c(HCl) = 0,1 mol/l. Dilute 8,28 ml of hydrochloric acid solution (4.7) to 1 l with water. SIST EN 15850:2010

technical grade. 4.14 Extraction solvent A. Mix 75 parts per volume of acetonitrile (4.11) with 25 parts per volume of water. 4.15 Injection solvent A for HPLC analysis. Mix four parts per volume of acetonitrile (4.11) with six parts per volume of water. 4.16 HPLC mobile phase A. Mix 53 parts per volume of acetonitrile (4.11) with 47 parts per volume of water. Filter and degas the HPLC mobile phase before use. 4.17 Extraction solvent B. Mix 75 parts per volume of methanol (4.13) with 25 parts per volume of water. 4.18 Washing solvent. Mix 15 parts per volume of methanol (4.12) with 85 parts per volume of PBS (4.10). 4.19 Injection solvent B for HPLC analysis. Mix five parts per volume of methanol (4.12) with five parts per volume of water. 4.20 HPLC mobile phase B. Mix 75 parts per volume of methanol (4.12) with 25 parts per volume of water. Filter and degas the HPLC mobile phase before use. SIST EN 15850:2010

bMA×××=ερ100maxzon (1) where Amax is the absorption determined at the maximum of the absorption curve (274 nm); M is the molar mass, in grams per mole, of zearalenone (M = 318,4 g/mol); 0 is the molar absorption coefficient, in square metres per mole of zearalenone in acetonitrile (4.11) ( 1 262 m2/mol, see [1]); b is the optical path length, in centimetres, of the quartz cell. Store this solution in a freezer at - 18 °C to - 20 °C. Allow to reach room temperature before opening. A solution stored in this way is usually stable for 12 months. Confirm the mass concentration of the solution if it is older than six months. 4.25 Zearalenone standard solution A,
= 2 µg/ml, for maize based baby food, barley flour, maize flour, polenta and wheat flour. Transfer an aliquot of the spiking solution (4.24) equivalent to 10 µg of zearalenone into either a vial (5.9) or a calibrated volumetric flask (5.10). Add acetonitrile (4.11) to make the total volume up to 5 ml.
...