Utilization of the totally synthesized structural analogues of Ochratoxin A in online monitoring of immunoaffinity columns capacity.

The growing concern over food contamination has driven the need for effective control and regulation of mycotoxins, making precise quantification of OTA levels critically important. Mycotoxin detection relies on immunoaffinity chromatography (IAC) for purification, recommended by AOAC, ISO, EN, and GB. However, IAC performance is limited by unstable immunoaffinity column capacity (IACC), affected by antibody quantity and activity. Disposable IACs are evaluated by batch sampling, failing to ensure individual consistency. Thus, online IACC monitoring technology is urgently required to guarantee detection accuracy and reliability. (84) RESULTS: The synthetic routes of six OTA structural analogues were designed starting from 2-iodo-6-methoxybenzoic acid in seven steps. Tyr-OTA-1 was screened and performed best owing to its property of the recovery rate of the IAC and the structure stability. Within the nominal column capacity range, the recovery rate of Tyr-OTA-1 gradually decreased when Tyr-OTA-1 was loaded in a certainly amount compared with OTA loading amount increased from low to high, while the recovery rate of OTA remained essentially unchanged at 88.72%-103.57% with a coefficient of variation ranging from 0.21% to 11.26%, indicating that the loading amount of Tyr-OTA-1 has no significantly inference on OTA detection. (104) SIGNIFICANCE: The analytical method based on marker Tyr-OTA-1 via IAC cascade HPLC in online monitoring of IACC was established, possessing the properties of high sensitivity, stability. Furthermore, this method exhibited good applicability for OTA detection in four cereal matrix samples (rice, millet, corn and wheat), demonstrating that Tyr-OTA-1 could achieve online monitoring of IACC. (53).