{"title":"Development of ELISA for Cyclopiazonic Acid in Various Foods and Its Validation Study Using LC/UV and LC/TOF-MS.","authors":"Koichi Saito, Ayako Kitora, Kana Hekizono, Maimi Kurata, Kazue Banba, Rie Ito, Hiroshi Akiyama","doi":"10.1093/jaoacint/qsaf088","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>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.</p><p><strong>Objective: </strong>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).</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>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.</p><p><strong>Highlights: </strong>The use of SPDE for CPA analyses resulted in effective cleanup, which subsequently enabled the application of ELISA for screening.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":""},"PeriodicalIF":1.7000,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of AOAC International","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1093/jaoacint/qsaf088","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
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.
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