Liquid Chromatography-Tandem Mass Spectrometry for Determination of Aflatoxin B1, Deoxynivalenol and Zearalenone in Artificial Porcine Gastrointestinal Digestive Juice

A rapid liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of aflatoxin B1 (AFB1), deoxynivalenol (DON) and zearalenone (ZEA) in artificial porcine gastrointestinal digestive juices, and successfully applied to the in vitro evaluation of adsorption efficiency of mycotoxin adsorbent. The formula feed was digested by artificial gastric and small intestinal juices in vitro, and then the mycotoxins and adsorbent were added in specific ratios. After incubation and centrifugation, the supernatant was diluted 10-fold and then analyzed by LC-MS/MS. The three mycotoxins were separated on a reversed-phase C18 column using a gradient elution program with 0.2 mM ammonium acetate aqueous solution and 0.1% formic acid in methanol as mobile phases. Qualitative analyses were performed under multiple-reaction monitoring mode, and quantitative analyses were carried out by using isotope internal standard method. Under the optimum conditions, the limit of quantification for AFB1, DON and ZEA was 1, 50 and 40 μg L−1 in gastric digested juice and 0.3, 50 and 20 μg L−1 in intestinal digested juice respectively, and the relative standard deviations (RSDs) were less than 5.0%. Then the thermal stability was investigated by incubating the analytes at 39.0 °C ± 0.5 °C for 1, 2, 5 and 10 h, and the experiment results showed that the three mycotoxins were stable under these conditions. Furthermore, the method was used to evaluate the binding efficacies of eight mineral adsorbents and five organic adsorbents. The mineral binders demonstrated binding efficacies of 85.13%–96.50%, 8.11%–14.71% and 13.67%–29.97% for AFB1, DON and ZEA in gastric digestive juice, and 76.15%–92.96%, 12.29%–31.31%, 0%–23.16% in intestinal digestive juice, respectively. The organic adsorbents exhibited binding efficacies of 7.42%–16.65%, 6.68%–16.24% and 18.56%–38.96% for AFB1, DON and ZEA in gastric digestive juice, and 8.65%–13.42%, 3.83%–23.49%, 24.88%–4.76% in intestinal digestive juice, respectively.

A rapid liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of aflatoxin B1, deoxynivalenol and zearalenone in artificial porcine gastrointestinal digestive juice. The isotope internal standard method was adopted to eliminate the interference of matrix effect. This method had successfully applied to the in vitro evaluation of adsorption efficiency of mycotoxin adsorbent.Figure optionsDownload full-size imageDownload as PowerPoint slide