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.