EN 14123:2007

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Lebensmittel - Bestimmung von Aflatoxin B1 und der Summe von Aflatoxin B1, B2, G1 und G2 in Haselnüssen, Erdnüssen, Pistazien, Feigen und Paprikapulver - Hochleistungsflüssigchromatographisches Verfahren mit Immunaffinitätssäulen-Reinigung und NachsäulenderivatisierungProduits alimentaires - Dosage de l'aflatoxine B1 et de la somme des aflatoxines B1, B2, G1 et G2 dans les noisettes, les cacahuètes, les pistaches, les figues et le paprika en poudre - Méthode par purification sur colonne d'immuno-affinité suivie d'une chromatographie liquide à haute performance avec dérivation post-colonneFoodstuffs - Determination of aflatoxin B1 and the sum of aflatoxin B1, B2, G1 and G2 in hazelnuts, peanuts, pistachios, figs, and paprika powder - High performance liquid chromatographic method with post-column derivatisation and immunoaffinity column cleanup67.080.01Sadje, 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 14123:2007SIST EN 14123:2008en,fr,de01-marec-2008SIST EN 14123:2008SLOVENSKI
STANDARDSIST EN 14123:20031DGRPHãþD

EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 14123
December 2007 ICS 67.050 Supersedes EN 14123:2003 English Version
Foodstuffs - Determination of aflatoxin B1 and the sum of aflatoxin B1, B2, G1 and G2 in hazelnuts, peanuts, pistachios, figs, and paprika powder - High performance liquid chromatographic method with post-column derivatisation and immunoaffinity column cleanup
Produits alimentaires - Dosage de l'aflatoxine B1 et de la somme des aflatoxines B1, B2, G1 et G2 dans les noisettes, les cacahuètes, les pistaches, les figues et le paprika en poudre - Méthode par purification sur colonne d'immuno-affinité suivie d'une chromatographie liquide à haute performance avec dérivation post-colonne
Lebensmittel - Bestimmung von Aflatoxin B1 und der Summe von Aflatoxin B1, B2, G1 und G2 in Haselnüssen, Erdnüssen, Pistazien, Feigen und Paprikapulver - Hochleistungsflüssigchromatographisches Verfahren mit Immunaffinitätssäulen-Reinigung und Nachsäulenderivatisierung This European Standard was approved by CEN on 12 November 2007.
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.
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Typical chromatograms.18 Annex B (informative)
Precision data.24 Bibliography.29
b0MAiii×××=100maxρ (1) where: Amax is the absorbance determined at the maximum of the absorption curve; Mi is the molar mass of each aflatoxin, in grams per mol; SIST EN 14123:2008

Aflatoxin Mi g/mol 0i
m2/mol B1 312 1930 B2 314 2040 G1 328 1660 G2 330 1790
4.21 Mixed aflatoxins stock solution Prepare a mixed aflatoxins stock solution containing 1000 ng/ml of aflatoxin B1 and G1, 200 ng/ml of aflatoxin B2 and G2 in the toluene and acetonitrile mixture (4.18) by appropriate dilution of aflatoxins (B1, B2, G1 and G2) stock solutions (4.20). NOTE A commercial total aflatoxins standard solution which is ready to use in a vial containing 1000 ng/ml of total aflatoxin may be used as an alternative. 4.22 Diluted mixed aflatoxins stock solution Prepare a diluted mixed aflatoxins stock solution containing 100 ng/ml of aflatoxin B1 and G1, 20 ng/ml of aflatoxin B2 and G2 in the toluene and acetonitrile mixture (4.18) by pipetting exactly 1,0 ml of the mixed aflatoxins stock solution (4.21) into a 10 ml calibrated volumetric flask (5.10), fill to the mark with the toluene and acetonitrile mixture (4.18) and mix well. Wrap the flask tightly in aluminium foil and store it at less than 4 °C or in a freezer. Before use, do not open the flask until the contents have reached room temperature to avoid incorporation of water by condensation. 4.23 Mixed aflatoxins calibration solutions Use the diluted mixed aflatoxins stock solution containing 100 ng/ml of aflatoxin B1 and G1, 20 ng/ml of aflatoxin B2 and G2 (see 4.22) for pipetting the volumes as given in Table 2 into a set of 10 ml volumetric flasks (5.10). Evaporate the toluene/acetonitrile solution just to dryness under a stream of nitrogen at room temperature. To each flask, add 4 ml of methanol, let aflatoxins dissolve, dilute to 10 ml with water, and shake well. Methanol and water are subject to volume contraction when mixed, so adjust the volume again to the given volume. SIST EN 14123:2008

Taken from diluted stock solution (4.22) µl B1
B2 G1 G2 1 40 0,400 0,080 0,400 0,080 2 120 1,200 0,240 1,200 0,240 3 200 2,000 0,400 2,000 0,400 4 280 2,800 0,560 2,800 0,560 5 360 3,600 0,720 3,600 0,720
4.24 Spiking solution Prepare a spiking solution by pipetting 2 ml of the mixed aflatoxins stock solution (containing 1000 ng/ml of aflatoxin B1 and G1, 200 ng/ml of aflatoxin B2 and G2, see 4.21) into a 10 ml calibrated volumetric flask. Evaporate the toluene/acetonitrile solution just to dryness under a stream of nitrogen at room temperature. Dilute to the mark with methanol and shake well. The concentration of this spiking solution is 200 ng/ml of aflatoxin B1 and G1, and 40 ng/ml of aflatoxin B2 and G2. Wrap the flask tightly in aluminium foil and store it at less than 4 °C. Before use, do not open the flask until the contents have reached room temperature to avoid incorporation of water by condensation. 5 Apparatus 5.1 General All glassware coming into contact with aqueous solutions of aflatoxins shall be washed with acid solution before use. Many laboratory washing machines do this as part of the washing program. Otherwise soak such laboratory glassware in sulfuric acid (2 mol/l) for several hours (e.g. 15 h overnight), then rinse well (e.g. three times) with water to remove all traces of acid. Check the absence of acid with pH paper. This treatment is necessary, because the use of non-acid washed glassware may cause losses of aflatoxins. In practice, the treatment is necessary for round bottomed flasks, volumetric flasks, measuring cylinders, vials or tubes used for calibration solutions and final extracts (particularly autosampler vials), and Pasteur pipettes, if these are used to transfer calibration solutions or extracts. 5.2 Usual laboratory apparatus and, in particular, the following 5.3 Laboratory mill, or explosion proof high speed blender1) , necessary for the production and extraction of pastes from hazelnuts, peanuts, pistachios and figs, with suitable blender jar 5.4 Adjustable vertical or horizontal shaker, needed for the analysis of paprika powder 5.5 Paper filter, e.g. 24 cm diameter, prefolded 5.6 Conical flask, with screw top or glass stopper
1) Contact your National Standardization Institute for appropriate high speed blenders. SIST EN 14123:2008

= 360 nm excitation filter and a wavelength of
= 420 nm cut-off emission filter, or equivalent (e.g. a detector with an adjustable monochromator). Recommended settings for adjustable detectors are 365 nm (excitation wavelength), 435 nm (emission wavelength) and a bandwidth of 18 nm. 5.12 Disposable filter unit, of pore size 0,45 µm Prior to usage, verify that no aflatoxin losses occur during filtration (recovery testing). NOTE There is a possibility that various filter materials can retain aflatoxins. 5.13 Pipettes, 2 ml, 5 ml and 10 ml capacity, with an accuracy of at least 0,5 % 5.14 Analytical balance, capable of weighing to 0,1 mg 5.15 Laboratory balance, capable of weighing to 0,01 g
2) KOBRA cell® is the trade name of a suitable product available commercially. 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 14123:2008
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