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An immunoaffinity column (IAC) for cyclopiazonic acid (CPA) was prepared by coupling a CPA-specific monoclonal antibody to CNBr-activated sepharose 4B. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was used to study the chromatographic behavior of a 0.2 ml_ gel column with a binding capacity of 4 μg CPA/column as well as to evaluate its efficacy as a cleanup tool for analysis of naturally occurring CPA. Sample extract either in buffer solution or in a solution containing up to 35% methanol could be loaded onto the column. After the column is washed with 5 ml_ deionized water and 5 ml_ 50% methanol, CPA could be quantitatively eluted with 2 ml_ 100% methanol. The column could be regenerated at least 10 times by washing with 10 mL equilibrating buffer and then storing in a cold room overnight before reuse. Recoveries of CPA added to corn, peanut, and mixed feed extracts in the range 10-200 ng/g were 88-105,86-100, and 90-110%, respectively. Detection limits were 2.0,4.4, and 4.7 ng/g for corn, mixed feed, and peanuts, respectively. Twenty-two peanut samples naturally contaminated with CPA were subjected to both IAC and solvent partition cleanup followed by dc-ELISA. Although a good correlation between data obtained from lAC-dc-ELISA and from SP-dc-ELISA (r = 0.75, p &amp;lt; 0.0001) was obtained, the slope of the linear regression was low (0.67), indicating loss during solvent partition cleanup. The overall data showed that the combination of IAC and dc- ELISA is an effective method for CPA analysis.

journal of aoac international

Immunoaffinity Column as Cleanup Tool for a Direct Competitive Enzyme-Linked Immunosorbent Assay of Cyclopiazonic Acid in Corn, Peanuts, and Mixed Feed