液相色谱-串联质谱法同时测定普洱茶中16种真菌毒素

为建立云南普洱茶中黄曲霉毒素、伏马毒素(FB₁-FB₃)、雪腐镰刀菌烯醇、脱氧雪腐镰刀菌烯醇(DON)、玉米赤霉烯酮、赭曲霉毒素A、T-2毒素、HT-2毒素以及杂色曲霉毒素等16种真菌毒素便捷准确的液相色谱-串联质谱(LC-MS/MS)检测方法,采用有机溶剂(甲醇-水-甲酸体积比为70∶29∶1)提取,通过PriboFast^®Multi-Toxin IAC免疫亲和净化柱净化,利用液相色谱串联质谱的多反应监测进行测定分析,采用内标法定量。结果表明,16种真菌毒素的回归方程均有良好的线性关系(r>0.99),3个不同加标水平下回收率为84.2%~102.7%,相对标准偏差为2.18%~4.78%。16种真菌毒素在174件云南普洱茶叶检测中,有53件茶叶检出DON,检出率为30.63%,96件茶叶检测出FB₁,检出率为55.17%,12件茶叶检测出FB₂,检出率为6.90%,12件茶叶中检测出FB₃,检出率为6.90%,其他真菌毒素均未检测出,且检出的4种真菌毒素含量在0.2~6.0 μg·kg^-1,均未超过国际食品安全规定的真菌毒素限量标准。说明云南省普洱茶叶中真菌毒素的检出率低,含量符合国家标准要求,该检测方法准确、可靠、便捷,适用于茶叶特别是批量茶叶中多种真菌毒素的检测。

Simultaneous detection and analysis of 16 kinds of mycotoxins in Pu-erh tea

To establish a convenient and accurate method for the determination of 16 mycotoxins with liquid chromatography-tandem mass spectrometry(LC-MS/MS) in Yunnan Pu-erh tea, including aflatoxin, fumonisin (FB₁-FB₂), nivalenol, deoxynivalenol (DON), zearalenone, ochratoxin A, T-2 toxin, HT-2 toxin and heterochromatoxin, samples were extracted by organic solvent (methanol-water-formic acid volume ratio was 70∶29∶1), purified by PriboFast^® Multi-Toxin IAC immunoaffinity purification column, determined and analyzed by multi-reaction monitoring of LC-MS/MS, and quantified by internal standard method. The regression equations of 16 mycotoxins had a good linear relationship(r>0.99). The recovery rate at three different levels was 84.2%-102.7%, and the relative standard deviation was 2.18%-4.78%. In the detection of 16 kinds of mycotoxins in 174 samples of Yunnan Pu'er tea, DON was detected in 53 samples of tea with the detection rate of 30.63%, FB₁ was detected in 96 samples of tea with the detection rate of 55.17%, and FB₂ was detected in 12 samples of tea with the detection rate of 6.90%, FB₃ was detected in 12 samples of tea with the detection rate of 6.90%. Other mycotoxins were not detected. The contents of the 4 mycotoxins detected were between 0.2-6.0 μg·kg^-1, which did not exceed the international food safety limit. The detection rate of mycotoxins in Pu-erh tea in Yunnan Province was low, and the content of mycotoxins met the requirements of national standards. The method was accurate, reliable and convenient, which can be applied to the detection of mycotoxins in tea, especially in batch tea.