液相色谱-三重四极杆质谱法测定花茶中5种吡咯里西啶类生物碱

Determination of 5 kinds of pyrrolizidine alkaloids in scented tea by liquid chromatography-triple quadrupole mass spectrometry

目的建立液相色谱-三重四极杆质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)同时测定花茶中5种吡咯里西啶类生物碱含量的分析方法。方法将花茶样品均质粉碎后,经0.1%甲酸水溶液浸泡,涡旋,超声提取,高速离心后取上清液过固相萃取柱,洗脱液氮吹吹干后用流动相复溶,经ZORBAX Eclipse PLUS C18色谱柱(2.1 mm×100 mm, 1.8μm)分离,以0.1%甲酸溶液和乙腈为流动相进行梯度洗脱,流速为0.30 m L/min,柱温为30℃,多反应监测模式测定,外标法定量。结果该方法线性关系良好,相关系数为0.997~0.998;方法的检出限(S/N=3)为3~5μg/kg、定量限(S/N=10)为10~15μg/kg;在15、30、150μg/kg添加水平下的平均回收率为89.69%~102.12%,相对标准偏差为0.3%~5.4%。结论该方法样品前处理简单快速、萃取效果好、灵敏度和选择性高,适用于常规检测。

Objective To establish a method for the determination of 5 kinds of pyrroliridine alkaloids in scented tea by liquid chromatography-tandem mass spectrometry(LC-MS/MS). Methods The sample was comminuted, and soaked in the aqueous solution of 0.1% formic acid, after vortex, ultrasonic extraction and high-speed centrifugation, and the supernatant was taken through the solid phase extraction column. After the eluting liquid nitrogen was blown and dried, the flow solution was re-dissolved. The samples were separated with a ZORBAX Eclipse PLUS C18 column(2.1 mm×100 mm, 1.8 μm) at the flow rate of 0.30 mL/min by gradient elution using 0.1% formic acid solution and acetonitrile as the mobile phase, and the column temperature was 30 ℃. The content of 5 kinds of pyrroliridine alkaloids was detected by multiple response monitoring model and quantitatively analyzed by external standard method. Results The linear relationship was good and the correlation coefficient was 0.997~0.998. The limit of detection of the method(S/N=3) was 3~5 μg/kg, the limit of quantitative(S/N=10) was 10~15 μg/kg, the average recovery rates at the addition levels of 15, 30, and 150 μg/kg were 89.69% to 102.12%, and the relative standard deviations were 0.3% to 5.4%. Conclusion This method is simple, rapid, effective, sensitive and selective, and suitable for conventional test.