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An analytical procedure for the determination of the Alternaria toxins radicinin, tenuazonic acid, altertoxin I, and alternariol methyl ether, in carrot and carrot-based products

The method is based on high performance liquid chromatography (HPLC) and UV diode array detection. A liquid chromatographic method for the determination of Alternaria radicina and A. alternata toxins in carrots was developed. Toxins were extracted from carrot with an acidified mixture of water, methanol and acetonitrile. The filtered extract was divided in two parts that were purified by solid phase extraction on a C18 column for the analysis of radicinin (RAD), altertoxin-I (ATX-I), alternariol (AOH), and alternariol methyl ether (AME) and on a polymeric Oasis HLB column for tenuazonic acid (TeA), respectively. Toxins were quantified by reversed phase liquid chromatography with ultraviolet diode array detector by using two consecutive isocratic mixtures of acetonitrile-sodium dihydrogen phosphate solution. Mean recoveries of TeA, ATX-I, AME, RAD, and AOH from carrots spiked at levels between 0.5 and 3.0µg/g were 69, 71, 90, 36, and 78%, with mean within-laboratory repeatability of 14, 5, 4, 6, and 18%, respectively. The mean between-laboratory reproducibilities for the determination of TeA, ATX-I, AME, and RAD in spiked samples were 25, 22, 6, and 12%, respectively. Limits of detection (signal-to-noise ratio of 3) for RAD, TeA, ATX-I, AME, and AOH were 0.006, 0.02, 0.02, 0.01, and 0.005µg/g, respectively. This method proved to be applicable to the analysis of 6-methoxymellein in carrot samples. This compound is a phytoalexin which is produced by the carrot itself in response to microbial infection or other form of stress and is associated with the bitterness of stressed carrots. For the analysis of carrot-based products (baby foods, carrot purees, puddings, sauces and vegetables soups) the method was appropriately modified. In particular, carrot-based products were diluted with water, centrifuged and the filtered extract purified by C18 column and analysed by reversed phase HPLC by using a gradient of acetonitrile in water as mobile phase. Detection limits were 0.02 and 0.01µg/g for carrots and carrot-based products, respectively.

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CNR Institute of Sciences of Food Production (ISPA)
Via Amendola 122/O
70125 BARI
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