血清中雷公藤甲素的液相色谱/质谱联用法测定研究

Determination of triptolide in serum by high-performance liquid chromatography mass spectrometry

目的:建立一种新型、灵敏、准确的人血清中雷公藤甲素的液相色谱/质谱联用测定方法。方法:血清经醋酸乙酯提取后,在Waters C18反相色谱柱(150 mm×3.9 mm i.d.,5μm)上,以醋酸-醋酸铵溶液(5 mmol/L)/乙腈/甲醇(60:30:10,v/v/v)为流动相,以泼尼松龙为内标,采用大气压化学电离(APCI)离子化方式在选择离子监测(SIM)模式下进行检测,定量检测离子为m/z[M-H]-359.1。结果:血清中雷公藤甲素测定的日内与日间精密度分别小于10.7%和11.3%,其在1.0～200.0 ng/ml范围具有良好的线性,检出限为1.0 ng/ml。结论:本方法可用于血清中痕量雷公藤甲素残留的测定。

Objective：To develop a novel,sensitive and accurate method for the determination of triptolide in serum samples by high-performance liquid chromatography（HPLC） coupled with an atmospheric pressure chemical ionization mass spectrometry（APCI-MS）.Methods：The separation was carried out on a Waters C18 reserved-phase column（150 mm×3.9 mm i.d.,5 μm） using acetic acid-acetic acid ammonium（5 mmol/L）/acetonitrile/methanol（60：30：10,v/v/v） as mobile phase in isocratic mode after serum samples were extracted by ethyl acetate.Detection was performed by an APCI-MS in the negative mode using prednisolone as an internal standard（IS）.The molecular ion m/z 359.1 -was selected for the quantification in selected ion monitoring（SIM） mode.Results：Within-day and day-to-day relative standard deviations were less than 10.7% and 11.3%,respectively.The linear range was 1.0-200.0 ng/ml for triptolide.The limit of quantification（LOQ） in serum samples was 1.0 ng/ml.Conclusion：The method can be applied to determination of trace triptolide in serum samples.