SIST EN 16187:2015

Foodstuffs - Determination of fumonisin B1 and fumonisin B2 in processed maize containing foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection after pre-column derivatisation

This European Standard specifies a method for the determination of fumonisin B1 (FB1) and fumonisin B2 (FB2) in processed maize-containing foods for infants and young children by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection (FLD). This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 112 µg/kg to 458 µg/kg for FB1+FB2, 89 µg/kg to 384 µg/kg for FB1 and 22 µg/kg to 74 µg/kg for FB2.
For further information on the validation, see Clause 8 and Annex B.

Lebensmittel - Bestimmung von Fumonisin B1 und Fumonisin B2 in Säuglings- und Kleinkindernahrung auf Maisbasis - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion nach Vorsäulenderivatiserung

Diese Europäische Norm legt ein Verfahren zur Bestimmung von Fumonisin B1 (FB1) und Fumonisin B2 (FB2) in Säuglings- und Kleinkindernahrung auf Maisbasis fest. Hierzu wird die Hochleistungsflüssig¬chromatographie (HPLC) mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion (FLD) eingesetzt. Dieses Verfahren wurde in einem Ringversuch durch die Untersuchung sowohl von natürlich kontaminierten Proben als auch von aufgestockten Proben mit Gehalten von 112 µg/kg bis 458 µg/kg für FB1  FB2, 89 µg/kg bis 384 µg/kg für FB1 und 22 µg/kg bis 74 µg/kg für FB2 validiert.
Weitere Informationen zur Validierung befinden sich in Abschnitt 8 und Anhang B.

Denrées alimentaires - Dosage de la fumonisine B1 et de la fumonisine B2 dans les aliments pour nourrissons et jeunes enfants contenant du maïs transformé - Méthode par CLHP avec purification sur colonne d'immunoaffinité et détection de fluorescence après dérivation précolonne

La présente Norme européenne spécifie une méthode de dosage de la fumonisine B1 (FB1) et de la fumonisine B2 (FB2) dans les aliments pour nourrissons et jeunes enfants contenant du maïs transformé par chromatographie liquide haute performance (CLHP) avec purification sur colonne d’immunoaffinité et détection par fluorescence (DFL). Cette méthode a été validée lors d’une étude interlaboratoires en procédant à l’analyse d’échantillons naturellement contaminés et dopés à des teneurs comprises entre 112 µg/kg et 458 µg/kg en FB1+FB2, entre 89 µg/kg et 384 µg/kg en FB1 et entre 22 µg/kg et 74 µg/kg en FB2.
Pour de plus amples informations sur la validation, voir l’Article 8 et l’Annexe B.

Živila - Določevanje fumonizina B1 in fumonizina B2 v predelani hrani na osnovi koruze za dojenčke in majhne otroke - Metoda HPLC s čiščenjem z imunoafinitetno kolono in fluorescenčno detekcijo po predkolonski derivatizaciji

Ta dokument določa metodo za določevanje fumonizina B1 in fumonizina B2 v predelani hrani na osnovi koruze za dojenčke in majhne otroke z metodo HPLC s čiščenjem z imunoafinitetno kolono in fluorescenčno detekcijo. Ta metoda je bila validirana v medlaboratorijski študiji z analizo naravno kontaminiranih in primešanih vzorcev od 112 µg/kg do 458 µg/kg za FB1+FB2, 89 µg/kg do 384 µg/kg za FB1 in 22 µg/kg do 74 µg/kg za FB2.
Za več informacij glede validacije glej točko 8 in dodatek B.

General Information

• Status: Published
• Publication Date: 30-Jul-2015
• ICS: 67.060 - Cereals, pulses and derived products
• ICS: 67.230 - Prepackaged and prepared foods
• Technical Committee: KŽP - Agricultural food products
• Current Stage: 6060 - National Implementation/Publication (Adopted Project)
• Start Date: 08-Jul-2015
• Due Date: 12-Sep-2015
• Completion Date: 31-Jul-2015
• Ref Project: EN 16187:2015 - Foodstuffs - Determination of fumonisin B1 and fumonisin B2 in processed maize containing foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection after pre-column derivatisation

Relations

• Replaces: SIST-TS CEN/TS 16187:2011 - Foodstuffs - Determination of fumonisin B1 and fumonisin B2 in processed maize containing foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection after precolumn derivatization
• Effective Date: 01-Sep-2015

Overview - EN 16187:2015 (CEN)

EN 16187:2015 defines a standardized analytical method for the determination of fumonisin B1 (FB1) and fumonisin B2 (FB2) in processed maize-containing foods for infants and young children. The method uses HPLC with immunoaffinity column (IAC) cleanup and fluorescence detection (FLD) after pre‑column derivatisation with o‑phthaldialdehyde (OPA). Validation details are provided in Clause 8 and Annex B; an interlaboratory study covered naturally contaminated and spiked samples (FB1+FB2: 112–458 µg/kg).

Key topics and technical requirements

• Scope: Quantitative determination of FB1 and FB2 in processed maize infant/young‑child foods.
• Sample preparation:
  • Homogenize (≤0.5 mm particle size); typical test portion 20 g.
  • Extraction with a citrate‑phosphate buffer:methanol:acetonitrile solvent (extraction solvent formulation specified).
  • Heat 55 °C for 60 min, shake, centrifuge (3 000 g, 15 min), filter and dilute prior to IAC.
• Immunoaffinity cleanup:
  • IAC must contain antibodies to FB1/FB2.
  • Column capacity ≥ 5 µg total fumonisins and recovery ≥ 80 % (when challenged with 5 µg).
• HPLC‑FLD analysis:
  • Pre‑column derivatisation with OPA reagent (with 2‑mercaptoethanol).
  • Reverse‑phase C18 column (e.g. 150 × 4.6 mm, 5 µm) with an appropriate binary gradient.
  • FLD settings typically around λex = 335 nm and λem = 440 nm.
• Calibration and working range:
  • Mixed FB1/FB2 certified standards and multi‑point calibration are used; working levels in the standard span sample equivalents up to ~480 µg/kg FB1 (see Clause 4.21 for calibration table).
• Safety: Fumonisins and solvents (acetonitrile, methanol) are hazardous - use fume hood and PPE.

Applications - who uses EN 16187:2015

• Food safety and regulatory laboratories for monitoring fumonisins in infant foods and demonstrating compliance.
• Quality control teams at infant food manufacturers and cereal processors.
• Contract testing laboratories conducting exposure assessments or routine batch testing.
• Research groups studying mycotoxin occurrence in processed maize products.
• Use cases include surveillance, batch release testing, contamination investigations and support for risk assessment.

Related standards

• EN 16187:2015 supersedes CEN/TS 16187:2011 (no technical change during conversion).
• Annex C provides a comparison with earlier fumonisin methods such as EN 14352:2004 and EN 13585:2001.
• Refer to Clause 8 and Annex B in EN 16187:2015 for full validation statistics from the interlaboratory study.

Keywords: EN 16187:2015, fumonisin B1, fumonisin B2, HPLC, immunoaffinity column, OPA pre‑column derivatisation, fluorescence detection, infant food testing, mycotoxin analysis.

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.PXQRDILQLWHWQRLebensmittel - Bestimmung von Fumonisin B1 und Fumonisin B2 in Säuglings- und Kleinkindernahrung auf Maisbasis - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion nach VorsäulenderivatiserungDenrées alimentaires - Dosage de la fumonisine B1 et de la fumonisine B2 dans les aliments pour nourrissons et jeunes enfants contenant du maïs transformé - Méthode par CLHP avec purification sur colonne d'immunoaffinité et détection de fluorescence après dérivation précolonneFoodstuffs - Determination of fumonisin B1 and fumonisin B2 in processed maize containing foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection after pre-column derivatisation67.230Predpakirana in pripravljena hranaPrepackaged and prepared foods67.060QMLKCereals, pulses and derived productsICS:Ta slovenski standard je istoveten z:EN 16187:2015SIST EN 16187:2015en,fr,de01-september-2015SIST EN 16187:2015SLOVENSKI
STANDARDSIST-TS CEN/TS 16187:20111DGRPHãþD

EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16187
June 2015 ICS 67.230 Supersedes CEN/TS 16187:2011English Version
Foodstuffs - Determination of fumonisin B1 and fumonisin B2 in processed maize containing foods for infants and young children - HPLC method with immunoaffinity column cleanup and fluorescence detection after pre-column derivatisation
Denrées alimentaires - Dosage de la fumonisine B1 et de la fumonisine B2 dans les aliments pour nourrissons et jeunes enfants contenant du maïs transformé - Méthode par CLHP avec purification sur colonne d'immunoaffinité et détection de fluorescence après dérivation précolonne
Lebensmittel - Bestimmung von Fumonisin B1 und Fumonisin B2 in Säuglings- und Kleinkindernahrung auf Maisbasis - HPLC-Verfahren mit Reinigung an einer Immunoaffinitätssäule und Fluoreszenzdetektion nach Vorsäulenderivatisierung This European Standard was approved by CEN on 7 May 2015.
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.
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Typical chromatograms . 14 Annex B (informative)
Precision data . 15 Annex C (informative)
Comparison between the method in this document and EN 14352:2004 and EN 13585:2001 on fumonisins in maize . 18 Bibliography . 19
FB1 and FB2a
FB1 µg/ml FB2 µg/ml FB1 µg/kg FB2 µg/kg 1 100 0,04 0,01 20,0 5,0 2 200 0,08 0,02 40,0 10,0 3 400 0,16 0,04 80,0 20,0 4 1 000 0,40 0,10 200,0 50,0 5 2 400 0,96 0,24 480,0 120,0 a For a sample extract reconstituted in 500 µl of the mixture of acetonitrile and water B (4.12).
1) Biopure RK 002003 and RK 002004 are examples of suitable products available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of these products. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 16187:2015

5.15 Heating block with nitrogen or air gas supply. 5.16 Vials, of 4 ml to 12 ml capacity with screw caps. 5.17 HPLC autosampler vials, of 1,8 ml capacity with caps. 5.18 Glass flat bottom vial insert, of 250 µl volume capacity. 5.19 Vortex mixer, or equivalent. 5.20 HPLC apparatus, comprising the following: 5.20.1 Injection system. 5.20.2 Mobile phase pump, (binary, ternary or quaternary pump), capable of generating a binary gradient at 1 ml/min. 5.20.3 Autosampler, capable of performing automated pre-column derivatisation with OPA reagent according to 6.4.1. 5.20.4 Analytical reverse-phase HPLC separating column, e.g. C18 reverse-phase column, 150 mm x 4,6 mm, 5 µm preceded by a suitable pre-column or guard filter, which provides acceptable retention and resolution for FB1 and FB2. SIST EN 16187:2015

2) Waters C18 SymmetryShield™ and Agilent Zorbax SB-C18 are examples of suitable products available commercially. This information is given for the convenience of users of this European Standard and does not constitute an end
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