Graduate Thesis Or Dissertation
 

Isolation and determination of aflatoxin in cottonseed meal and other feedstuffs

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  • The aflatoxins, carcinogenic and toxic mold metabolites of Aspergillus flavus, were isolated and determined in cottonseed meal and other feedstuffs. This new quantitative method uses an acetone soxhlet extraction to remove the toxins from the defatted meal. The residual triglycerides, phospholipids and pigments were removed from the acetone by filtration of the cooled solution. The aflatoxin was further isolated by evaporating the acetone and redissolving the residue in hot methanol. After cooling, the insolubles were discarded and the methanol-soluble materials taken up in chloroform. The chloroform solution was spotted on silicic acid thin layer chromatographic plates along with suitable standards. After development with 9:1 (v/v) chloroform:acetone, the chromatographic plates were examined under ultraviolet light. Complete recovery of aflatoxin, within limits of visual discrimination, was obtained by this isolation procedure as indicated by extraction efficiency and internal standard data. Twenty-five cottonseed meals were analyzed by this method and nine meals contained aflatoxin B₁ at levels from 17 to 190 parts per billion. Fluorodensitometry, a new instrumental technique, was used to compare standards and samples containing aflatoxin directly from the thin-layer chromatographic plate. This procedure eliminates the errors inherent in the visual comparison method and permits greater sensitivity and accuracy. Amounts of aflatoxin B₁ as low as 8.0 x 10⁻⁵ micrograms can be detected by this technique. The results obtained by this procedure were substantiated by duckling assay and trout feeding trials.
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