EN 14333-2:2004

Non fatty foods - Determination of benzimidazole fungicides carbendazim, thiabendazole and benomyl (as carbendazim) - Part 2: HPLC method with gel permeation chromatography clean up

This draft European Standard specifies a high performance liquid chromatographic method for the determination of the benzimidazole fungicides carbendazim and thiabendazole in fruits, vegetables and processed products.
When benomyl is present, it is completely degraded to carbendazim and is also determined as carbendazim. Thiophanate-methyl is partly decomposed and therefore not quantitatively determined.
The method has been validated for carbendazim and thiabendazole in an interlaboratory test with homogenates of apples, French beans, mushrooms, lemons and fruit based infant food.

Fettarme Lebensmittel - Bestimmung der Benzimidazol-Fungizide Carbendazim, Thiabendazol und Benomyl (als Carbendazim) - Teil 2: HPLC-Verfahren mit Reinigung durch Gelpermeationschromatographie

Diese Europäische Norm legt ein hochleistungs-flüssigchromatographisches Verfahren zur Bestimmung der Benzimidazol-Fungizide Carbendazim und Thiabendazol in Obst, Gemüse und deren Verarbeitungsprodukten fest.
Wenn Benomyl anwesend ist, wird es vollständig zu Carbendazim abgebaut und ebenfalls als Carbendazim bestimmt. Thiophanat-methyl wird teilweise zersetzt und deshalb nicht quantitativ bestimmt.
Das Verfahren ist für Carbendazim und Thiabendazol durch einen Ringversuch mit Homogenaten von Äpfeln, grünen Bohnen, Pilzen, Zitronen und Kindernahrung aus Fruchtzutaten validiert worden.

Aliments non gras - Détermination des benzimidazoles antifongiques, le carbendazime, le thiabendazole et le bénomyl en tant que carbendazime - Partie 2: Méthode CLHP avec purification par chromatographie par perméation de gel

Le présent projet de Norme européenne décrit une méthode de chromatographie en phase liquide à haute performance pour la détermination du carbendazime et du thiabendazole, benzimidazoles antifongiques, dans les fruits, les légumes et les produits transformés.
Si du bénomyl est présent, celui-ci est complètement dégradé en carbendazime et dosé en tant que carbendazime. Le thiophanate-méthyle est partiellement décomposé et ne peut, par conséquent, être déterminé quantitativement.
La présente méthode a été validée pour le carbendazime et le thiabendazole lors d'un essai interlaboratoires réalisé à l'aide de broyats de pommes, de haricots verts, de champignons, de citrons et d'aliments pour bébé à base de citrons et de fruits.

Nemastna živila - Določevanje benzimidazolnih fungicidov: karbendazim, tiabendazol in benomil (kot karbendazim) - 2. del: Metoda HPLC s čiščenjem z gelsko kromatografijo

General Information

• Status: Withdrawn
• Publication Date: 05-Oct-2004
• Withdrawal Date: 31-Dec-2014
• ICS: 67.080.01 - Fruits, vegetables and derived products in general
• Technical Committee: CEN/TC 275 - Food analysis - Horizontal methods
• Drafting Committee: CEN/TC 275/WG 4 - Pesticides in foods of plant origin
• Current Stage: 9960 - Withdrawal effective - Withdrawal
• Start Date: 01-Jan-2015
• Completion Date: 01-Jan-2015
• Ref Project: SIST EN 14333-2:2005 - Non fatty foods - Determination of benzimidazole fungicides carbendazim, thiabendazole and benomyl (as carbendazim) - Part 2: HPLC method with gel permeation chromatography clean up

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.D]LPFettarme Lebensmittel - Bestimmung der Benzimidazol-Fungizide Carbendazim, Thiabendazol und Benomyl (als Carbendazim) - Teil 2: HPLC-Verfahren mit Reinigung durch GelpermeationschromatographieAliments non gras - Détermination des benzimidazoles antifongiques, le carbendazime, le thiabendazole et le bénomyl en tant que carbendazime - Partie 2: Méthode CLHP avec purification par chromatographie par perméation de gelNon fatty foods - Determination of benzimidazole fungicides carbendazim, thiabendazole and benomyl (as carbendazim) - Part 2: HPLC method with gel permeation chromatography clean up67.080.01Sadje, zelenjava in njuni proizvodi na splošnoFruits, vegetables and derived products in general67.050Splošne preskusne in analizne metode za živilske proizvodeGeneral methods of tests and analysis for food productsICS:Ta slovenski standard je istoveten z:EN 14333-2:2004SIST EN 14333-2:2005en01-januar-2005SIST EN 14333-2:2005SLOVENSKI
STANDARD
EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 14333-2October 2004ICS 67.080.01 English versionNon fatty foods - Determination of benzimidazole fungicidescarbendazim, thiabendazole and benomyl (as carbendazim) -Part 2: HPLC method with gel permeation chromatography cleanupAliments non gras - Détermination des benzimidazolesantifongiques, le carbendazime, le thiabendazole et lebénomyl en tant que carbendazime - Partie 2: MéthodeCLHP avec purification par chromatographie parperméation de gelFettarme Lebensmittel - Bestimmung der Benzimidazol-Fungizide Carbendazim, Thiabendazol und Benomyl (alsCarbendazim) - Teil 2: HPLC-Verfahren mit Reinigungdurch GelpermeationschromatographieThis European Standard was approved by CEN on 29 July 2004.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 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 translationunder the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia,Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2004 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 14333-2:2004: ESIST EN 14333-2:2005

Precision data.10 Annex B (informative)
Alternative GPC column.13 Annex C (informative)
HPLC with reversed-phase column.14 Bibliography.15
(NaOH) = 26 g/100 ml 3.7 Diluted sodium hydroxide solution,
(NaOH) = 2,6 g/100 ml 3.8 GPC eluting mixture: Cyclohexane (3.4) / ethyl acetate (3.3) 1 + 1 (V/V) 3.9 Solvent mixture: dilute 5 ml ethyl acetate (3.3) with cyclohexane (3.4) to 100 ml 3.10 Mobile phase A for HPLC: Cyclohexane (3.4) 3.11 Mobile phase B for HPLC: Cyclohexane (3.4) / isopropanol / methanol 85 + 15 + 5 (V/V/V) To 250 ml of this mixture, add one drop of ammonia solution (25 g/100 g). Prior to use, filter the mixture through a membrane filter (4.7). SIST EN 14333-2:2005

(carbendazim) = 25 mg/100 ml in methanol 3.14 Thiabendazole stock solution,
(thiabendazole) = 65 mg/100 ml in acetone 3.15 Standard solutions Dilute the carbendazim stock solution (3.13) or the thiabendazole stock solution (3.14) with ethyl acetate (3.3) and cyclohexane (3.4) to obtain appropriate dilutions in cyclohexane / ethyl acetate of about 80 + 20 (V/V) to 75 + 25 (V/V). When injected into the HPLC system (4.6), the dilutions should contain less than 10 ml/100 ml of methanol and less than 2 ml/100 ml of acetone. 4 Apparatus 4.1 General Usual laboratory equipment and in particular the following :
4.2 Food chopper 4.3 High speed blender or homogenizer 4.4 Rotary evaporator with a water bath 4.5 Instrument for GPC equipped with glass column with two adjustable end pieces, 500 mm long, 10 mm inner diameter, and with at least one sample loop (1 ml), column packing BioBeads®1) S-X3 resin, pre-swelled overnight in the GPC eluting mixture (3.8) and packed as described in
5.3.1. NOTE Apparatus for automated GPC is commercially available. 4.6 High performance liquid chromatograph equipped with 4.6.1 Pumping system, with three solvent reservoirs, an injection valve for 15 µl, a UV detector and a fluorescence detector connected in series and a quantification unit with an integrating system; 4.6.2 HPLC analytical column, stainless steel cartridge, 150 mm long, 4,6 mm inner diameter, packed with a suitable diol-bonded silica for normal phase chromatography, particle size 3 µm. 4.7 Membrane filter, pore size 0,45 µm, suitable for organic solutions 4.8 Glass fibre filter, 90 mm diameter 4.9 Syringe filter, pore size 0,45 µm, suitable for organic solutions
1) BioBeads ® is a trade name of a product supplied by Bio-Rad Laboratories, Richmond, CA, USA. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of the product named. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 14333-2:2005

5.2.2 Lemons, limes, plums Proceed as described in 5.2.1, but add 6,0 ml sodium hydroxide solution (3.6) instead of 3,0 ml. 5.2.3 Juices Check which volume (x ml) of diluted sodium hydroxide solution (3.7) is required to adjust a portion of 7,5 g of the juice to approximately pH 10. Weigh a test portion of 75 g (m) to the nearest 0,5 g into the jar of a blender (4.3). Add 200 ml (V1) of ethyl acetate (3.3) and x ml of sodium hydroxide solution (3.6) and homogenize the mixture for 30 s. Proceed as described in 5.2.1. 5.3 Gel permeation chromatography 5.3.1 Packing gel permeation column Allow approximately 9 g of BioBeads S-X3 resin (4.5) to swell overnight in the GPC eluting mixture (3.8). Then pour the suspension all at once into the column. As soon as the gel bed has settled (free from air bubbles) to a level of approximately 40 cm, insert the plunger, lower it down to the bed level and screw it into place. Start the flow of GPC eluting mixture at a low rate and gradually increase it to 1 ml/min. If the gel bed sinks to a still lower level, adjust the plunger accordingly (observe manufacturer’s instructions). NOTE For alternative GPC column, see Annex B. 5.3.2 Checking elution volumes Load the sample loop with 1 ml of appropriately diluted standard solutions in cyclohexane (3.4) / ethyl acetate (3.3) 1 + 1 (V/V), elute as described in 5.3.3 and determine, whether carbendazim and thiabendazole are completely recovered. Collect fractions of 0,5 ml around the expected starting and end points of the elution range. NOTE In general, the elution range is 20 ml to 27 ml for carbendazim and 24 ml to 30 ml for thiabendazole. SIST EN 14333-2:2005
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