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By Mary W. Trucksess, Albert E. Pohland

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Close the vial with a cap and mix the solution well. 5 min in a 65°C water bath (the level of water must be above the level of solution in the vial). 6. Inject 20 µL of solution on liquid chromatograph (LC) with the fluorescence detector set at 360 nm excitation and 440 nm emission. The column is 15 cm C18 and the mobile phase is acetonitrile-methanol-water (1 + 1 + 4). Chromatogram should have only one peak (see Note 6). 1. 2. 3. 4. 4. Preparation of Vials of Aflatoxin Dry Film Standards 1. Determine the repeatability of the adjustable dispenser, 0 to 2 mL, by weighing 10 × 1 mL portions of toluene-acetonitrile (9 + 1) into tared flasks, recording the weight of each portion.

Integrate the area of each protonated molecule peak (see Note 14). 3. Total the area of the protonated molecules for all compounds, the main component (here FB1) plus all impurities (see Note 15). 4. Total the area of the main component only (see Note 16). 5. Divide the numerical value from step 4 by that from step 3, multiply by 100 to give the percent purity of the FB1. 4. Notes 1. For analyses using a parallel ELSD, one would omit scrubbing distilled water with the organic sieve. The sieve often contaminates the water (and sample blanks) with resins which, although they contain no UV-chromophore and hence are not observed by UV detection, would nevertheless give a strong background signal in an ELSD.

Isolation of Mycotoxins 25 3 Preparatory Isolation of Mycotoxins from Solid Phase Fungal Cultures Robert M. Eppley 1. Introduction Mycotoxins are a large group of secondary fungal metabolites possessing significantly different chemical and physical properties. Because of this diversity, no general procedures can be developed for the isolation and purification of all of the different mycotoxins. The aim of this chapter is to present a procedure that has been successfully applied to the isolation and purification of one mycotoxin group (the fumonisins) which can be adapted to the preparation of many of the other mycotoxins.

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