EN 14524:2004

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.þLWYLMRLebensmittel - Bestimmung von Okadasäure in Muscheln - HPLC-Verfahren mit Reinigung durch Festphasenextraktion, Derivatisierung und fluorimetrischer BestimmungProduits alimentaires - Dosage de l'acide okadaique dans les moules - Méthode par CLHP avec purification par extraction sur phase solide, dérivation et détection fluorimétriqueFoodstuffs - Determination of okadaic acid in mussels - HPLC method with solid phase extraction clean-up, derivatization and fluorimetric detection67.120.30Ribe in ribji proizvodiFish and fishery productsICS:Ta slovenski standard je istoveten z:EN 14524:2004SIST EN 14524:2005en01-januar-2005SIST EN 14524:2005SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 14524
August 2004 ICS 67.120.30
English version
Foodstuffs - Determination of okadaic acid in mussels - HPLC method with solid phase extraction clean-up, derivatization and fluorimetric detection
Produits alimentaires - Dosage de l'acide okadaïque dans les moules - Méthode par CLHP avec purification par extraction sur phase solide, dérivation et détection fluorimétrique
Lebensmittel - Bestimmung von Okadasäure in Muscheln -HPLC-Verfahren mit Reinigung durch Festphasenextraktion, Derivatisierung und fluorimetrischer Bestimmung This European Standard was approved by CEN on 21 May 2004.
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 Central Secretariat 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 Central Secretariat has the same status as the official versions.
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Typical chromatogram.12 Annex B (informative)
Precision data.14 Bibliography.15
4.7 Ethyl acetate 4.8 Dichloromethane, stabilised with 2-methyl-2-butene 4.9 Methanol
4.10 Methanol solution, 3 = 80 %
4.11 n-hexane
4.12 9-anthryldiazomethane (ADAM) Weigh solid ADAM in discrete portions (e.g. 1 mg), store at ≤ -18 °C and dissolve in ethyl acetate just before use. 4.13 Derivatization solution, mass concentration
= 1,5 g/l in ethyl acetate (4.7) 4.14 Solvent mixture of n-hexane and dichloromethane Mix 1 part per volume of n-hexane (4.11) with 1 part per volume of dichloromethane (4.8). 4.15 Solvent mixture of dichloromethane and acetone Mix 9 parts per volume of dichloromethane (4.8) with 1 part per volume of acetone (4.4). 4.16 Solvent mixture of acetonitrile and dichloromethane Mix 5 parts per volume of acetonitrile (4.5) with 1 part per volume of dichloromethane (4.8). 4.17 Mixture of methanol and ethyl acetate Mix 1 part per volume of methanol (4.9) with 1 part per volume of ethyl acetate (4.7). 4.18 HPLC mobile phase solvent A Mix 800 ml of methanol (4.9) with 200 ml of ethyl acetate (4.7). 4.19 HPLC mobile phase solvent B Mix 700 ml of acetonitrile (4.5) with 300 ml of water. SIST EN 14524:2005

Dilute okadaic acid methylanthrylester with methanol (4.9) to a concentration of 0,1 µg/ml. This corresponds to 1 600 µg/kg mussel hepatopancreas (in samples). Store the solution in a brown bottle. At ≤ -18 °C the solution is stable for at least 6 months. 4.24 Mussels free of okadaic acid and related compounds 5 Apparatus 5.1 General Usual laboratory apparatus, and in particular: 5.2 Instrument for separation of hepatopancreas, e.g. scalpel 5.3 Blender or homogenizer 5.4 Centrifuge, capable of a centrifugal force up to 3 000 g, with suitable tubes, e.g. 20 ml 5.5 Pear shaped flask, e.g. 10 ml and 25 ml capacity 5.6 Glass sample tubes with stoppers, e.g. 10 ml capacity 5.7 One-mark volumetric flask, 20 ml capacity 5.8 Analytical balance, accurate to the nearest 0,1 mg 5.9 Solid phase extraction (SPE) cartridges, (3 ml) filled with 650 mg silica gel with solvent reservoir (approx. 10 ml), commercially available 5.10 Suitable pipettes of various volumina
1) This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN. Equivalent products may be used if they can be shown to lead to the same results.
–18 °C has no negative influence on the result of this method. 7 Procedure 7.1 Preparation of test sample Transfer fresh mussels into boiling water and cook for 10 min. Separate 20 g to 30 g of hepatopancreas material from approximately 1 kg of mussels (including the shells). The hepatopancreas is the green or brown part of the mussel. If only smaller sample amounts of hepatopancreas material are available, the whole material has to be prepared. Homogenize the hepatopancreas material with the homogenizer or blender (5.3) and start immediately with the preparation or otherwise store the homogenate at –18 °C. SIST EN 14524:2005
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