Analysis of Fumonisins and Alternaria alternata Toxin by Liquid Chromatography-Enzyme-Linked Immunosorbent Assay

Use of a direct competitive enzyme-linked immunosorbent assay (ELISA) as a postcolumn monitoring system after liquid chromatography (LC) is described for analysis of different fumonisin analogs. Without cleanup and derivatization, sample extracts are directly injected into a C18 reversed-phase column and then subjected to LC. Fractions (0.5 mL each) are collected and then analyzed by ELISA. LC using a water-methanol gradient separated the 3 major fumonisins FmB1, FmB2, and FmB3, and as low as 0.1 ng FmB1 could be detected. Recovery of FmB1 added to ground corn (100-1000 ng/g) and then extracted with CH3CN-H2O (1 + 1, v/v) was 78.8%. Analysis of fumonisins in one starch and 14 naturally contaminated corn samples showed that FmB1 was the major fumonisin. Ten samples also were contaminated with FmB2, but only 2 samples were contaminated with FmB3. The method also was used to analyze extracts from cultures of 3 Alternaria alternate (AAL) strains. Both FmB1 and the AAL toxin TA were detected in the culture extracts, and their amounts varied considerably with the cultures tested.