同位素内标-超高效液相色谱-串联质谱法检测花生及其制品中黄曲霉毒素B1及其生物可给性研究

Determination of aflatoxin B1 in peanuts and its products by isotope internal standard-ultra performance liquid chromatography-tandem mass spectrometry and study on its bioaccessibility

中文关键词:  花生  同位素内标法  超高效液相色谱-串联质谱法  黄曲霉毒素B1  生物可给性

英文关键词:peanut  isotope internal standard method  ultra performance liquid chromatography-tandem mass spectrometry  aflatoxin B1  bioaccessibility

中文摘要:

      目的 建立一种基于同位素内标的超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)定量分析花生及其制品中的黄曲霉毒素B1 (aflatoxin B1, AFB1), 并研究其在胃、肠两个阶段的生物可给性。方法 样品经乙腈-水-乙酸(84:15:1, V:V:V)提取, 高速冷冻离心净化后, 以0.1%甲酸溶液与甲醇为流动相, 经Waters Acquity BEH C18柱(100 mm×2.1 mm, 1.7 μm)分离, 内标法定量。通过体外模拟消化实验, 研究样品中AFB1在胃、肠两个阶段的生物可给性。结果 AFB1在0.01~30.00 μg/L范围内线性关系良好, 相关系数(r2)为0.9998, 花生、花生油、花生酱、花生奶中AFB1的检出限为0.07~0.36 μg/kg、定量限为0.24~1.20 μg/kg, 3种加标水平的平均回收率为80.3%~103.4%、相对标准偏差均小于5.8%; 市售样品中, 花生相对于其制品的AFB1污染更严重, 2份超过国家限量标准(20 μg/kg); 花生类食品中AFB1在胃相中的生物可给性为4.35%~33.88%、在胃肠相中为12.81%~79.78%。结论 本研究所建立的检测方法简单快速、精密度高、准确性好, 可以满足对花生类食品中AFB1的快速准确分析的要求, 体外模拟消化表明不同花生类食品中AFB1生物可给性存在较大差异。

英文摘要:

      Objective To establish a method for the quantitative analysis of aflatoxin B1 (AFB1) in peanuts and its products by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) based on isotope internal standard method, and study its bioaccessibility in the gastric and the small intestinal phases. Methods The sample was extracted with acetonitrile-water-acetic acid (84:15:1, V:V:V), purified by high-speed refrigerated centrifugation, and separated on a Waters Acquity BEH C18 column (100 mm×2.1 mm, 1.7 μm) with 0.1% formic acid solution and methanol as mobile phases, the isotope internal standard method was used for quantitative analysis. On this basis, the bioaccessibility of AFB1 in the gastric and the small intestinal phases was studied by simulated digestion experiment in vitro. Results The AFB1 had good linear relationship in range of 0.01-30.00 μg/L with the correlation coefficient (r2) of 0.9998, the limits of detection and the limits of quantification were from 0.07? 0.36 μg/kg and from 0.24?1.20 μg/kg, respectively, the average recoveries of the 3 spiked levels ranged from 80.3% to 103.4% with the relative standard deviations all less than 5.8%; Among the commercial samples, the AFB1 contamination of peanuts was more serious than that of its products, and 2 of them exceeded the national limit standard (20 μg/kg). The bioavailability of AFB1 in peanut foods ranged from 4.35%?33.88% in the gastric phase and 12.81%?79.78% in the small intestinal phase. Conclusion The method established in this study is simple, rapid, with high precise and accurate, which can meet the requirements the rapid and accurate analysis of AFB1 in peanut foods. In vitro simulated digestion shows that the bioaccessibility of AFB1 in different peanut foods is quite different.