Development of ELISA for Cyclopiazonic Acid in Various Foods and Its Validation Study Using LC/UV and LC/TOF-MS.

Background: Food contamination with mycotoxins has become a concern in recent years. Cyclopiazonic acid (CPA) is a mycotoxin found in foods, such as peanuts, corn, and cheese, which poses health hazards to humans.

Objective: To develop and validate an enzyme-linked immunosorbent assay (ELISA) for detecting CPA in various food matrices using appropriate pretreatment methods, with liquid chromatography/ultraviolet detection (LC/UV) and/or LC/time of flight mass spectrometry (TOF-MS).

Methods: Pretreatment of food was optimized using liquid-liquid extraction and solid-phase dispersive extraction (SPDE), and validated across liquid and solid foods. Indirect competitive ELISA was performed. The data from recovery experiments were analyzed using a one-way analysis of variance.

Results: All food specimens added at high concentrations yielded satisfactory ELISA results and exhibited a strong correlation with the LC/UV measurements. In the case of moderate-concentration additions, satisfactory accuracy was obtained for foods other than spices. At low concentrations, the results of ELISA and instrumental analyses (LC/UV and LC-TOF/MS) were comparable for sake, soy sauce, noodle soup, corn, and miso. However, elevated values were observed for the ELISA of cheese, peanuts, and spices.

Conclusion: Although the low recovery levels of CPA in spices suggests the need for a more effective cleanup method, the developed ELISA is suitable as a rapid screening method for CPA in various foods, with LC confirmation recommended in complex matrices.

Highlights: The use of SPDE for CPA analyses resulted in effective cleanup, which subsequently enabled the application of ELISA for screening.