同位素内标-超高效液相色谱-三重四极杆质谱法测定谷物中3种A型单端孢霉烯毒素

Determination of 3 kinds of type A trichothecenes in cereals by isotope internal standard-ultra performance liquid chromatography-triple quadrupole mass spectrometry

中文关键词:  A型单端孢霉烯毒素  谷物  同位素内标  超高效液相色谱-三重四极杆质谱

英文关键词:type A trichothecenes  cereals  isotope internal standard  ultra performance liquid chromatography-triple quadrupole mass spectrometry

中文摘要:

      目的 建立同位素内标-超高效液相色谱-三重四极杆质谱法同时测定谷物中T-2毒素、HT-2毒素和二乙酰镳草镰刀菌烯醇(diacetoxyscirpenol, DAS) 3种A型单端孢霉烯毒素(trichothecenes, TCTs)的方法。方法 待测样品中加入同位素内标后用含1%甲酸的乙腈水溶液(乙腈:水:甲酸=70:29:1, V:V:V)提取, 用5 mmol/L乙酸铵溶液和含5 mmol/L乙酸铵-0.1%甲酸的甲醇作为流动相进行梯度洗脱, 经超高效液相色谱-三重四极杆质谱法的多反应监测模式(电喷雾正离子模式)测定, 采用同位素内标法定量。结果 该方法中DAS在0.05~2.00 ng/mL范围内、T-2毒素和HT-2毒素在0.10~2.00 ng/mL范围内线性关系良好, 相关系数r均大于0.999。检出限为0.10~0.20 μg/kg, 定量限为0.30~0.60 μg/kg, 回收率为81.0%~119.3%, 精密度良好, 相对标准偏差(relative standard deviation, RSD)低于10% (n=3)。结论 该方法简单、快速、灵敏、准确, 适用于谷物中3种A型TCTs的同时定量测定。

英文摘要:

      Objective To establish a method for simultaneous determination of 3 kinds of type A trichothecenes (TCTs), including T-2 toxin, HT-2 toxin and diacetoxyscirpenol (DAS) in cereals by isotope internal standard-ultra performance liquid chromatography-triple quadrupole mass spectrometry. Methods The samples were spiked with isotope internal standards and extracted with acetonitrile-water-formic acid (70:29:1, V:V:V). Chromatographic separation was achieved through gradient elution using 5 mmol/L ammonium acetate solution and methanol containing 5 mmol/L ammonium acetate-0.1% formic acid as mobile phases. Detection was performed in multiple reaction monitoring mode with positive electrospray ionization by ultra performance liquid chromatography-triple quadrupole mass spectrometry, and quantification was conducted using the isotope internal standard method. Results The method demonstrated excellent linearity with correlation coefficients (r)>0.999 for DAS (0.05–2.00 ng/mL) and T-2/HT-2 toxins (0.10–2.00 ng/mL). The limits of detection and quantification ranged 0.10–0.20 μg/kg and 0.30–0.60 μg/kg, respectively. The recoveries ranged from 81.0% to 119.3%, and the precision was good, with the relative standard deviations (RSDs) were below 10% (n=3). Conclusion This developed method exhibits simplicity, rapidity, sensitivity and accuracy, making it suitable for simultaneous quantitative determination of 3 kinds of type A TCTs in cereals.