Fusarium fungi are capable of producing, to a variable degree, two or more toxins. The major Fusarium mycotoxins are fumonisins, A- and B-trichothecenes, and zearalenone (ZON) (1). Trichothecenes are responsible for a wide range of toxicity in animals, including feed refusal, weight loss and vomiting. In particular deoxynivalenol (DON) can inhibit protein biosynthesis and has been reported as an immunosuppressant (2). To reduce the levels of biogenic toxins, European authorities are currently discussing further regulations on mycotoxins. Within the European Union (EU), harmonized legislation is setting maximum limits for aflatoxins and ochratoxin A in cereals and cereal products. Limits for Fusarium toxins (DON, ZEA, HT2, and T2) are currently being drafted in EU member states; for example, maximum limits for Fusarium toxins (DON 100–500 μg/kg, ZON 20–50 μg/kg) were established in February 2004 (3).

The most common hyphenated methods for the determination of A- and B-trichothecenes include gas chromatography–electron capture detection, gas chromatography–mass spectrometry (GC–MS) or liquid chromatography (LC)–postcolumn derivatization and fluorescence detection (4). Increasingly, LC–tandem MS (MS-MS) has been applied to mycotoxin analysis despite higher costs and the need for experienced personnel. The main advantages of the technique include its general applicability to a broad range of compounds, high sensitivity and outstanding selectivity. Several methods already have been reported for the simultaneous determination of mycotoxins, which offer significant advantages over conventional techniques (5–9).

Here, we present a new method for the analysis of mycotoxins in cereal-based samples using a triple quadrupole LC–MS-MS system (API 2000, Applied Biosystems, Foster City, California). The method analyzes the mycotoxins deoxynivalenol (DON), nivalenol (NIV), fusarenone X (FX), verrucarol (VOL), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), diacetoxyscirpenol (DAS), HT-2 toxin (HT2), T-2 toxin (T2), zeralanone (ZAN), and zearalenone (ZON). For additional verification purposes ochratoxin A and aflatoxins were included in the study. We will briefly discuss our experiences regarding the choice of solvents used, solvent flow, split of LC eluents, negative or positive ionization and a comparison of APCI and ESI interfaces.

Figure 1: Standard chromatogram of the Fusarium mycotoxins of major interest.