CEN/TC 275/WG 5 - Biotoxins

This document describes a method for the determination of T-2 toxin and HT-2 toxin in cereals and cereal-based products, e.g. oats, intended for nutrition of infants and young children by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) after cleanup by solid phase extraction (SPE) [5].
The method has been validated for HT-2 toxin in oat flour at levels of 9,3 µg/kg and 28,1 µg/kg, oat flakes at levels of 16,5 µg/kg and 21,4 µg/kg, and breakfast cereals (containing oat flakes) at a level of 8,1 µg/kg and for T-2 toxin in oat flour at levels of 4,4 µg/kg and 8,3 µg/kg, oat flakes at levels of 4,9 µg/kg and 6,6 µg/kg and breakfast cereals (containing oat flakes) at a level of 3,5 µg/kg.
Laboratory experiences [6] have shown that the method is also applicable to highly swelling materials (dry cereal-based porridges and modified starches), but these were not examined in the method validation study. Details are outlined in 7.3.
The method can also be applied to oat-by-products at higher levels of T-2- and HT-2 toxin. In this case, the dilution steps need to be considered [6].
The method can also be applied to cereals and cereal products for infants and young children based on e.g. wheat, barley and rice. In this case, the method needs to be in-house-validated for each material. At the time of the interlaboratory study, planned range was 10 µg/kg to 100 µg/kg, and it is known from the pre-study that the method works well in the whole range, although final validation was only done in the range from 3,5 µg/kg to 28,1 µg/kg.

This document describes a method using isotopically labelled standards for the quantitative determination of aflatoxins B1, B2, G1, G2 and M1 (AFB1, AFB2, AFG1, AFG2 and AFM1), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEN), T-2 and HT-2 toxins (T-2 and HT-2) and fumonisins B1 and B2 (FB1 and FB2) in foods by liquid chromatography (LC) coupled with tandem mass spectrometry (MS/MS).
A specific immunoaffinity column (IAC) clean-up is needed for aflatoxins (AFs) and OTA in food intended for infants and young children (e.g. infant cereals, milk-based powders), in spices, in dried fruits and in nuts.
The method has been validated through an intercollaborative study on different commodity groups: cereals and cereal-based products including food for infant and young children, nuts, spices, dried fruits and milk powder. The measuring range of each mycotoxin in these naturally contaminated and/or spiked food samples were:
- AFB1: 0,085 7 µg/kg - 11,4 µg/kg;
- AFB2: 0,079 2 µg/kg - 12,5 µg/kg;
- AFG1: 0,062 8 µg/kg - 20,9 µg/kg;
- AFG2: 0,052 0 µg/kg - 15,0 µg/kg;
- AFM1: 0,034 2 µg/kg - 0,110 µg/kg;
- OTA: 0,448 µg/kg - 17,2 µg/kg;
- DON: 45,2 µg/kg - 743 µg/kg;
- ZEN: 9,57 µg/kg - 131 µg/kg;
- T-2: 10,3 µg/kg - 57,9 µg/kg;
- HT-2: 9,50 µg/kg - 81,8 µg/kg;
- FB1: 31,1 µg/kg - 4 260 µg/kg;
- FB2: 44,2 µg/kg - 1 300 µg/kg.
The measuring ranges of the method for each mycotoxin/matrix combination are given in Table 8.

This European Standard specifies a method for the determination of five Alternaria toxins in wheat, tomato juice and sunflower seed samples by liquid chromatography tandem mass spectrometry (LC-MS/MS). The method includes the analysis of Altenuene (ALT), Alternariol (AOH), Alternariol monomethyl ether (AME) in the range of 1 μg/kg to 100 μg/kg, and Tentoxin (TEN) in the range of 5 μg/kg to 500 μg/kg, and Tenuazonic acid (TEA) in the range of 10 μg/kg to 1000 μg/kg.

This document describes a procedure for the determination of the citrinin content in food (cereals, red yeast rice (RYR)), herbs and food supplements by liquid chromatography tandem mass spectrometry (LC-MS/MS).
This method has been validated for citrinin in red yeast rice and in the formulated food supplements in the range of 2,5 μg/kg to 3 000 μg/kg and in wheat flour in the range of 2,5 μg/kg to 100 μg/kg.
Laboratory experiences have shown that this method is also applicable to white rice, herbs such as a powder of ginkgo biloba leaves and the formulated food supplements in the range of 2,5 μg/kg to 50 μg/kg.

This document describes a method for the determination of the sum of six ergot alkaloids (ergocornine, ergometrine, ergocristine, ergotamine, ergosine and ergocryptine) and their -inine epimer pairs by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) after clean-up by dispersive solid phase extraction (dSPE).
The method has been validated in the range 13,2 µg/kg to 168 µg/kg for the sum of the twelve ergot alkaloids, in rye flour, rye bread and cereal products (breakfast cereal, infant breakfast cereal, and crispbread) that contained rye as an ingredient, as well as seeded wholemeal flour and a barley and rye flour mixture.
Method performance was satisfactory in the range 24,1 µg/kg to 168 µg/kg, however at lower concentrations RSDR values were greater than 44 %, and HorRat values exceeded 2,0, indicating the method may not be fully suitable at concentrations below 24 µg/kg for sum of ergot alkaloids, although it is suitable for screening at these concentrations.

This document describes a procedure for the determination of aflatoxins B1, B2, G1 and G2 and total aflatoxins (sum of B1, B2, G1 and G2) in spices for which EU maximum levels are established, other than paprika, by high performance liquid chromatography (HPLC) with post-column derivatization (PCD) and fluorescence detection (FLD) after immunoaffinity column (IAC) clean-up.
The method is applicable to the spices capsicum (excluding paprika), pepper, nutmeg, ginger, turmeric and mixtures thereof.
The method has been validated for aflatoxins B1, B2, G1 and G2 and total aflatoxins in a range of test samples that comprised: ginger, pepper, nutmeg, chilli, turmeric as individual spices and mixed pepper + chilli + nutmeg (90 + 5 + 5, m + m + m), mixed spice+ginger (6 + 4, m + m) mixed spice, mixed turmeric+ginger (2 + 8, m + m).
The validation was carried out over the following concentration ranges: aflatoxin B1 = 1 µg/kg to 16 µg/kg and total aflatoxins = 2,46 µg/kg to 36,1 µg/kg.

This document specifies a procedure for the determination of ochratoxin A (OTA) in chilli, paprika, black and white pepper, nutmeg, spice mix, liquorice (root and extracts), cocoa and cocoa products by high performance liquid chromatography (HPLC) with immunoaffinity column clean-up and fluorescence detection (FLD).
This method has been validated in interlaboratory studies via the analysis of both naturally contaminated and spiked samples ranging from 1,0 μg/kg to 84,9 μg/kg for spices (paprika and chili [5], black and white pepper, nutmeg and spice mix [6]), ranging from 7,7 μg/kg to 96,8 μg/kg for liquorice and liquorice products [7] and ranging from 2,1 μg/kg to 26,3 μg/kg for cocoa and cocoa products [6].
For further information on the validation, see Clause 10 and Annex B.

This document describes a procedure for the determination of ochratoxin A (OTA) in pork products specifically ham, pork-based products (canned chopped pork) and pork liver using high performance liquid chromatography with fluorescence detection (HPLC-FLD).
The method has been validated for ochratoxin A in naturally contaminated ham, pork based products (canned chopped pork) and pork liver containing 0,5 μg/kg to 11 μg/kg [4], [5], [6].
Laboratory experiences have shown that this method is also applicable to pâté and kidney [4].

This document specifies a procedure for the determination of phomopsin A in lupin seeds and lupin-derived products based on liquid chromatography with tandem mass spectrometry (LC-MS/MS). Several phomopsins exist, i.e. phomopsin A, B, C and D, but the method only deals with the quantitative measurement of phomopsin A due to lack of commercially available analytical reference standards for the other phomopsins.
The method has been validated for phomopsin A in naturally contaminated lupin seeds, lupin flour and crisp bread at levels ranging from approximately 5 µg/kg to 60 µg/kg

This document describes a screening method for the determination of aflatoxin B1, deoxynivalenol, fumonisin B1 and B2, ochratoxin A, HT-2 and T-2 toxins, and zearalenone in foodstuffs by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS).
The aim of the screening method is to test compliance of foodstuff with regulatory limits or to determine whether a certain pre-defined level (the screening target concentration, STC) is exceeded or not. The result of the screening is either "negative" or "suspect". "Negative" (screen negative) means that the targeted mycotoxins are not detected or potentially present but below the STC. "Suspect" (screen positive) means that the established cut-off level is exceeded and the sample can contain one or more mycotoxins at a level higher than the STC.
For full identification and accurate quantification a second confirmatory quantitative analysis method is required which is outside the scope of this document.
The method is suitable for various types of foodstuff and has been validated for representative matrices from four commodity groups:
-   high starch and/or protein content and low water and fat content: wheat, cereal mixture, wheat flour and cornflakes;
-   high oil content: peanuts;
-   high sugar low water content: figs;
-   high water content: grape juice.
During validation, cut-off levels were established for the following screening target concentrations:
-   aflatoxin B1: 2 µg/kg to 5 µg/kg;
-   deoxynivalenol: 250 µg/kg to 865 µg/kg;
-   fumonisin B1: 200 µg/kg to 790 µg/kg;
-   fumonisin B2: 110 µg/kg to 230 µg/kg;
-   ochratoxin A: 4 µg/kg to 9 µg/kg;
-   T-2 toxin: 25 µg/kg;
-   HT-2 toxin: 25 µg/kg to 50 µg/kg;
-   zearalenone: 30 µg/kg to 100 µg/kg.

This document specifies a procedure for the determination of nivalenol (NIV), deoxynivalenol (DON) and its acetyl derivatives (3-acetyl-DON and 15-acetyl-DON), HT-2 and T-2 toxins (HT-2 and T-2) and zearalenone (ZEN) in cereals and cereal products by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) after clean-up by solid phase extraction (SPE).
The method has been validated with samples of wheat, wheat flour, and wheat crackers. The wheat and the wheat flour was prepared from a mixture of wheat and fungi infected wheat kernels. The wheat crackers were baked from wheat flour and water spiked with the target mycotoxins.
Validation levels for NIV ranged from 27,7 μg/kg to 378 μg/kg.
Validation levels for DON ranged from 234 μg/kg to 2420 μg/kg.
Validation levels for 3-acetyl-DON ranged from 18,5 μg/kg to 137 μg/kg.
Validation levels for 15-acetyl-DON ranged from 11,4 μg/kg to 142 μg/kg.
Validation levels for HT-2 ranged from 6,6 μg/kg to 134 μg/kg.
Validation levels for T-2 ranged from 2,1 μg/kg to 37,6 μg/kg.
Validation levels for ZEN ranged from 31,6 μg/kg to 230 μg/kg.
Laboratory experiences have shown that this method is also applicable to barley and oat flour, and rye based crackers [5], however, this has not been validated in a collaborative study.

This European Standard describes a procedure for the determination of the zearalenone content in edible vegetable oils specifically maize germ oil by either of the following techniques: High performance liquid chromatography with fluorescence detection (LC-FLD) or high performance liquid chromatography with tandem mass spectrometry (LC-MS/MS) after basic extraction of the diluted oil.
The method has been validated for zearalenone in naturally contaminated maize germ oil at levels of 61,2 µg/kg to 515 µg/kg [5].
Laboratory experiences [6] have shown that this method is also applicable to other vegetable oils such as wheat germ oil (n = 4), sunflower oil (n = 5), pumpkin seed oil (n = 1), soybean oil (n = 5), hemp seed oil (n = 5), rape seed oil (n = 11), and mixed oils including maize germ oil (n = 3). However occasionally, samples can result in interferences in the FLD-chromatograms. In this case, the detection with MS/MS is recommended.

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.

ISO 16050:2003 specifies a reverse-phase high-performance liquid chromatographic method, with immunoaffinity column clean-up and post-column derivatization, for the determination of aflatoxins in cereals, nuts and derived products. The limit of quantification for aflatoxin B1, and for the sum of aflatoxins B1, B2, G1 and G2, is 8 micrograms per kilogram.
The method has been validated for maize containing 24,5 micrograms per kilogram, for peanut butter containing 8,4 micrograms per kilogram, and for raw peanuts containing 16 micrograms per kilogram of total aflatoxins. It has also been shown that this method can be used for oilseed products, dried fruits and derived products.

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.

This European Standard specifies a method for the determination of aflatoxin B1 in baby food by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection. This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 0,07 µg/kg to 0,18 µg/kg.
For further information on the validation, see Clause 9 and Annex B.

This Technical Report gives criteria for single laboratory validated methods of analysis for the determination of mycotoxins. The criteria and topics covered are accuracy, trueness, recovery, precision, measurement uncertainty, selectivity, applicability, linearity, limit of detection, limit of quantification, sensitivity, ruggedness, specificity. This report also contains information on terms and definitions, validation, standardization procedures and interlaboratory studies by international organizations (e.g. AOAC, CEN, ISO, IUPAC, IDF). Confirmatory methods and screening methods are described. The validation criteria specified for mycotoxins in general are given.

This European Standard specifies a method for the determination of deoxynivalenol (DON) in cereals (grain and flour), cereal based foods and cereal based foods for infants and young children by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and UV detection. This method has been validated in three interlaboratory studies. The first study was for the analysis of samples of wheat, rice flour, oat flour, maize, polenta, and wheat based breakfast cereal ranging from 85,4 µg/kg to 1 768 µg/kg, the second study was for wheat and maize ranging from 165 µg/kg to 4 700 µg/kg and the third study was for cereal based foods for infants and young children ranging from 58 µg/kg to 452 µg/kg.
For further information on the validation, see Clause 9 and Annex B.
WARNING — The use of this standard can involve hazardous materials, operations and equipment. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

This European Standard specifies a method for the determination of patulin in fruit juices and fruit-based purée, such as baby food purée, using high performance liquid chromatography with ultra-violet detection (HPLC-UV). Using naturally contaminated and spiked samples this method has been validated for the determination of patulin in apple juice, at levels ranging from 3,0 μg/kg to 15,5 μg/kg, and in fruit-based baby food purée, at levels ranging from 3,4 μg/kg to 17,9 μg/kg. Baby food fruit purée (commercially available on the European market) used in this study contained the following ingredients: blueberry; apple; banana; lemon; wheat biscuits; wheat syrup; whole milk; and vegetable oil. A detailed listing, including the fractions, of each product used in this study is given in [1].
Further information on validation, see Clause 9 and Annex B.

This European Standard specifies a method for the determination of ochratoxin A in cereal based foods for infants and young children by high performance liquid chromatography (HPLC) with immunoaffinity column cleanup and fluorescence detection. This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 0,050 µg/kg to 0,217 µg/kg. For further information on the validation see Clause 8 and Annex B. Additional studies have shown that this method is applicable to cereal based baby foods containing 8 different types of cereals, honey and cocoa, at levels up to 3,540 µg/kg, see Annex C and [6].

This European Standard specifies a method for the determination of ochratoxin A in currants, raisins, sultanas, mixed dried fruit and dried figs by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection. This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 1,1 µg/kg to 11 µg/kg.
For further information on the validation, see Clause 9 and Annex B.
WARNING - The use of this standard can involve hazardous materials, operations and equipment. This standard does not purport to address all the safety problems associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

This European Standard specifies a method for the determination of ochratoxin A content in wine and beer using immunoaffinity column clean up and high performance liquid chromatography (HPLC), see [2] and [3]. This method has been validated in an interlaboratory study according to AOAC International Guidelines [4] for collaborative study procedures to validate characteristics of a method of analysis for the determination of ochratoxin A in wine and beer via the analysis of naturally contaminated and spiked samples of wine and beer at
levels ranging from 0,1 ng/ml to 3 ng/ml.

This European Standard specifies a method for the determination of ochratoxin A content in barley and roasted coffee using immunoaffinity column clean up and high performance liquid chromatography (HPLC). This method has been validated for ochratoxin A contents in barley in the range from 0,1 g/kg up to 4,5 g/kg and for roasted coffee in the range from 0,2 g/kg up to 5,5 g/kg.

This European Standard is applicable to the determination of aflatoxins B1, B2, G1 and G2 in hazelnuts, figs, pistachios, peanuts and paprika powder. The limit of quantification of the method is 0,8 ng/g for each aflatoxin or better (value derived from in-house and collaborative study), depending on the equipment used.

A comparison was made between dry milling and slurry mixing as comminution step preceding mycotoxins analysis. Such in respect to EC legislation that consists of sample schemes up to 30 kg. Cacao, green coffee, almonds and pistachio samples of 10 kg were milled by a RAS mill and all three sub-samples were completely analysed for aflatoxin B1 or Ochratoxin A. The differences in analytical results are explained by measurements of particle size distributions of both milling types. The obtained data are compared with literature data on coefficients of variation (CV) for various milling procedures. For dry milling CV values were generally not below 20 % for aflatoxin B1 levels up to 38 µg/kg in peanuts, whereas slurry mixing could achieve CV values below 5 % at aflatoxin B1 levels down to 4 µg/kg in pistachios. Measurements also showed possible difference in mycotoxin content of a sample between both milling types. This could lead to false positive or negative results when rejecting or accepting a lot, as this is based on the sample result. It was concluded that slurries contain smaller particles than dry milled samples and thus generate the lowest possible CV values which in turn leads to better sample homogenisation.

This document specifies a method for the detemination of fumonisin B1 (FB1) and fumonisin B2 (FB2) in maize based foods using high performance liquid chromatography (HPLC) and immunoaffinity clean-up, see [1], [2], [3].
The method has been successfully validated in a collaborative study according to AOAC Guidelines for collaborative study procedures [4] to validate characteristics of a method of analysis for the determination of fumonisins in maize flour and corn flakes containing 323 µg/kg to 1414 µg/kg FB1 and 90 µg/kg to 558 µg/kg FB2.

This European Standard specifies a method for the determination of patulin in apple juices and apple puree up to 128 ug/kg using high performance liquid chromatography (HPLC).

This European Standard describes a method for the determination of T-2 toxin and HT-2 toxin in cereals and cereal based products e.g. oats, intended for nutrition of infants and young children by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) after cleanup by solid phase extraction (SPE) [5].
The method has been validated for HT-2 toxin in oat flour at levels of 9,3 µg/kg and 28,1 µg/kg, oat flakes at levels of 16,5 µg/kg and 21,4 µg/kg, and breakfast cereals (containing oat flakes) at a level of 8,1 µg/kg and for T-2 toxin in oat flour at levels of 4,4 µg/kg and 8,3 µg/kg, oat flakes at levels of 4,9 µg/kg and 6,6 µg/kg and breakfast cereals (containing oat flakes) at a level of 3,5 µg/kg.
Laboratory experiences [6] have shown that the method is also applicable to highly swelling materials (dry cereal based porridges and modified starches), but these were not examined in the method validation study. Details are outlined in 6.3.
The method can also be applied to oat-by-products at higher levels of T-2- and HT-2 toxin. In this case, the dilution steps need to be considered [6].
The method can also be applied to cereals and cereal products for infants and young children based on e.g. wheat, barley, and rice. In this case, the method needs to be in-house-validated for each material. At the time of the interlaboratory study, planned range was 10 µg/kg to 100 µg/kg, and it is known from the pre-study that the method works well in the whole range, although final validation was only done in the range from 3,5 µg/kg to 28,1 µg/kg.

This document describes a procedure for the determination of the citrinin content in food (cereals, red yeast rice (RYR)), herbs and food supplements by liquid chromatography tandem mass spectrometry (LC MS/MS).
This method has been validated for citrinin in red yeast rice and in the formulated food supplements in the range of 2,5 µg/kg to 3000 µg/kg and in wheat flour in the range of 2,5 µg/kg to 100 µg/kg.
Laboratory experiences have shown that this method is also applicable to white rice, herbs such as a powder of ginkgo biloba leaves and the formulated food supplements in the range of 2,5 µg/kg to 50 µg/kg.

This CEN Report gives criteria for the selection of methods of analysis for mycotoxins. Criteria covered are the repeatability, reproducibility, recovery, extraction solvents, applicability and food types. Performance criteria are included for aflatoxin B1, total of aflatoxins B1, B2, G1 and G2, aflatoxin M1, ochratoxin A, patulin, fumonisin B1, fumonisin B2, deoxynivalenol, nivalenol, HT-2 toxin, T-2 toxin and zearalenone.

This European Standard specifies a method for the determination of ochratoxin A (OTA) of greater than 3 microgramme/kg. The method has been successfully validated in interlaboratory studies according to ISO 5725:1986 (1) on whole barley containing 2,9 microgramme/kg and 3,0 microgramme/kg, 7,4 microgramme/kg and 14,4 microgramme/kg of ochratoxin A, on whole maize containing 8,2 microgramme/ kg and 16,3 microgramme/kg of ochratoxin A as well as on wheat bran containing 3,8 microgramme/kg and 4,5 microgramme/kg of ochratoxin A. Note: Numerous laboratory experiences have shown that this method is also applicable to wheat flour.

This European Standard specifies a method for the determination of ochratoxin A of greater than 0,4 ug/kg. The method has been successfully validated in 2 interlaboratory studies according to ISO 5725:1996 (1) on wheat whole meal containing 0,4 microgramme/kg and 1,2 microgramme/kg of ochratoxin A. Note: Numerous laboratory experiences have shown that this method is also applicable to cereals, dried fruits, oilseeds, pulses, wine, beer, fruit juices and raw coffee, see (2), (3), (4).

This European Standard specifies a method for the determination of fumonisin B1 (FB1) and fumonisin B2 (FB2) in maize using high performance liquid chromatography (HPLC).
The method has been successfully validated in an interlaboratory study according to AOAC Guidelines for Collaborative Studies [1] on maize containing 405 µg/kg to 6732 mg/kg fumonisin B1 and 152 mg/kg to 2619 mg/kg fumonisin B2. The method works well with maize or minimally processed maize (e.g. fresh, dried and milled maize), but does not provide reliable results with most maize-based processed products.

This Technical Specification 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.

This European Standard specifies a method for the determination of aflatoxin contents of greater than 8 ug/kg. The method has been successfully validated in an interlaboratory study according to ISO 5725:1986 on maize containing 24,5 ug/kg,peanut butter containing 8,4 ug/kg and raw peanuts containing 16 ug/kg of total aflatoxins. Some laboratory experiences have shown that this method can be used to several types of cereals, oilseed products, shell-fruits, dried fruits and derived products, after in-house validation.

This European Standard specifies a method for the quantitative determination of the content of okadaic acid in mussels and mussel products. The content of okadaic acid is determined as free extractable acid of mussel hepatopancreas. Okadaic acid, a fat-soluble toxin from dinophysis algae, is a main component of dinophysis toxins.
The method has been validated in an interlaboratory study according to ISO general principles on assessing accuracy of measurement methods and results. The limit of determination of this method (signal/noise = 10) is 100 µg/kg for okadaic acid in mussel hepatopancreas. The method has been validated for okadaic acid in cooked mussels at levels of 441 µg/kg to 1 467 µg/kg.
Laboratory experiences have shown that this method can also be used to determine other dinophysis toxins, e.g. dinophysis toxins 1, 2 and 3 (DTX-1, DTX-2 and DTX-3), see [1], [2], [3], [4], [5] and [6]

TC - Modification to Equation 1

TC - Modification to Equation 1

This European Standard specifies a method for the determination of ochratoxin A content in wine and beer using immunoaffinity column clean up and high performance liquid chromatography (HPLC), see [2], [3].
This method has been validated in an interlaboratory study according to AOAC International Guidelines [4] for collaborative study procedures to validate characteristics of a method of analysis for the determination of ochratoxin A in wine and beer via the analysis of naturally contaminated and spiked samples of wine and beer at levels ranging from 0,1 ng/ml to 3 ng/ml.

This European Standard specifies a method for the determination of ochratoxin A content in barley up to 6 ug/kg and ochratoxin A content in roasted coffee up to 6 ug/kg using immunoaffinity (IA) column clean up and high performance liquid chromatography (HPLC).

This draft European Standard is applicable to the determination of aflatoxins B1, B2, G1 and G2 in figs, pistachios, peanuts and paprika powder. The limit of quantification of the method is 0,8 ng/g for each aflatoxin or better (value derived from in-house and collaborative study), depending on the equipment used.