Enzyme-linked immunosorbent assay of aflatoxin B1 in naturally contaminated corn and cottonseed.

Naturally contaminated corn and cottonseed samples were screened for aflatoxin B1 (AFB1) by a direct competitive enzyme-linked immunosorbent assay (ELISA). Samples were blended 5 min in an extraction solvent of methanol-water-dimethylformamide (70 + 29 + 1) and filtered. Filtrates were assayed by direct competition between AFB1 in the corn and cottonseed samples and AFB1-peroxidase conjugate for binding to specific antibody adsorbed to a solid phase microtiter plate. Standard curves prepared using the extract of AFB1-free corn and cottonseed samples, and extraction solvent only, showed negligible interference by the sample extract in the performance of ELISA. The AFB1 content in corn and dehulled cottonseed samples as determined by ELISA ranged from 7 to 422 micrograms/kg and 7 to 3,258 micrograms/kg, respectively. When ELISA estimates of AFB1 in corn were compared with values obtained by thin layer chromatography (CB method), the correlation coefficient (n = 10) was 0.95. Average interassay and subsample coefficients of variation for ELISA in corn were 21.4 and 22.0%, respectively. When ELISA estimates of AFB1 in cottonseed were compared with values obtained by liquid chromatography (Pons method), the correlation coefficient (n = 15) was 0.96. Using this ELISA, 36 duplicate sample extracts can be screened for AFB1 in less than 2 h.

Treatment of freshly harvested 1980 Georgia dent corn samples collected for aflatoxin analysis.

In 1980, corn was harvested from six 15-ft rows in each of 67 fields in Georgia for aflatoxin analysis. Every sixth ear from each field was placed in a sample bag to be dried the day of collection. The rest of the corn was husked and shipped to Peoria in cardboard boxes. When undried ear samples arrived in Peoria, each sample was randomly separated into 5 equivalent subsamples. One set of 67 subsamples was shelled and dried as soon as possible to avoid further aflatoxin formation. Two other sets of 67 subsamples were stored 3 and 6 weeks before shelling and drying. The remaining 2 sets of ear samples were placed in plastic bags with 5% Monoprop (1 part propionic acid plus 1 part versite) and stored 3 and 6 weeks before shelling and drying. The samples dried in Georgia before shipping had an average total aflatoxin level of 217 ng/g. Samples shelled and dried immediately after arrival had an average level of 202 ng/g. Samples shelled and dried after 3 and 6 weeks of storage had average total aflatoxin levels of 417 and 387 ng/g, respectively. Samples stored 3 and 6 weeks in the presence of 5% Monoprop (2.5% propionic acid) had average total aflatoxin levels of 120 and 157 ng/g, respectively.