Aflatoxin (AF) is the strongest know naturally occurring carcinogen. It can be present in various foods, spices or medicinal plants. No legal limits have yet been provided for medicinal herbs and there are not available standards method for its assay, although it is well known that some plants could be frequently contaminated.

The method of Reif and Metzger (1995) was adopted and it was optimized for determination of aflatoxins in herbal drugs from Poland. The modified method has three major steps. The first, the samples were extracted with 70 % (v/v) methanol in water and centrifuged. The supernatant was then filtered through a folded filter paper. The second, the clear filtrate was diluted with 10 % (v/v) Tween20 / PBS and cleaned up with an immunoaffinity column (IAC). After washing the IAC with wather, the aflatoxins were eluted from the column with methanol. The final step was the separation and determination of AF by reversed-phase LC and detected by fluorescence after post column derivatization (PCD) involving bromination. PCD was achieved with an electrochemical (Kobra) cell by addition of bromide to the mobile phase. The extraction procedure was optimized in order to obtain the best recovery and minor changes to HPLC conditions were intoduced to improve the method. The use of twice larger volume of extraction solvent and addition of a surfactant improved aflatoxins recovery.

Statistical analysis of the data was performed to determine recoveries and repeatability of the method. Recoveries of AFs (B1, B2, G1) added to cortex salicis and semen lini were over 70% at two levels of fortification (higher level: 2 ng/g for every type of AFs, lower level (only cortex): 0.5 ng/g, for every type of AFs except for the case of B1 in semen lini (68%). The recoveries of G2 were 34-37%. For cortex salix, relative standard deviations for repeatability (RSD) for all aflatoxins ranged from 1. 4 to 5% for two contamination levels. For semen RSD values for all aflatoxins ranged 0. 4 to 3. 8 %.

This validated assay was applied to determine the degree of contamination of AFs in 16 various herbal medicinal products currently marketed in Poland. The method was successfully carried out with all herbal products diversified as to compositions and dosage forms. The results revealed that ten of herbal samples were contaminated and in two samples detectable amount of the total AFs was higher then the current legislative level for foods. The assay will be continued.

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