超高效液相色谱高分辨质谱法同时测定蜂蜜中的5种雷公藤类物质

Simultaneous determination of 5 kinds of tripterygium substances in honey by liquid chromatography high resolution mass spectrometry

目的建立超高效液相色谱高分辨质谱法(UPLC-Q-Exactive)同时测定蜂蜜中5种雷公藤类物质(雷公藤甲素、雷公藤红素、雷公藤吉碱、雷公藤次碱、去甲泽拉木醛)。方法蜂蜜加水稀释后经乙腈提取,以HSS T3色谱柱(2.1 mm×100 mm,1.8μm)分离,甲醇-5 mmol/L乙酸铵+0.1%甲酸水溶液为流动相梯度洗脱,高分辨质谱仪检测,外标法定量。结果雷公藤甲素、雷公藤红素、雷公藤吉碱和雷公藤次碱的线性范围为1μg/L~40μg/L、去甲泽拉木醛的线性范围为5μg/L~200μg/L;线性相关系数r为0.999 1~0.999 9;检出限分别为0.01 mg/kg、0.01 mg/kg、0.01 mg/kg、0.01 mg/kg、0.05 mg/kg;加标回收率为96.1%~104.2%,精密度为1.26%~6.52%。结论 Q-Exactive技术定性准确,四级杆与Orbitrap相结合提升了检测的灵敏度,溶剂效应及基质效应的影响很低。本方法线性范围宽,准确度好,精密度高,能满足对蜂蜜中5种雷公藤类物质的定性定量检测,且前处理步骤少,操作简单,适用于蜂蜜中毒样品的快速测定。

Objective To establish a method for the simultaneous determination of 5 kinds of tripterygium substances in honey( including triptolide,tripterine,wilforgine,wilforine,demethylzeylasteral) by ultra high performance liquid chromatography-high resolution mass spectrometry( UPLC-Q-Exactive). Methods Honey was diluted with water and extracted with acetonitrile,separated by HSS T3 column( 2. 1 mm × 100 mm,1. 8 μm),and detected by high resolution mass spectrometer and quantified by the external standard. The methanol-5 mmol/L ammonium acetate + 0. 1% formic acid aqueous solution was used as mobile phase gradient elution. Results The linear correlation coefficient( r) was within 0. 999 1-0. 999 9. When the concentrations of triptolide,tripterine,wilforgine,and wilforine were within 1 μg/L-40 μg/L,and the concentration of demethylzeylasteral was within 5 μg/L-200 μg/L. The detection limits are 0. 01 mg/kg,0. 01 mg/kg,0. 01 mg/kg,0. 01 mg/kg,0. 05 mg/kg,the recoveries ranged from 96. 1% to 104. 2%,and the precision from 1. 26% to 6. 52%.Conclusion Q-Exactive technology is qualitatively accurate. The combination of quadrupole and orbitrap improves the detection sensitivity,and the effects of solvent and matrix are low. This method has a wide linear range,accuracy,and precision. It can be used for the qualitative and quantitative detection of 5 kinds of tripterygium substances in honey. The method is simple,fast,and it is suitable for the rapid determination of honey poisoning samples.