超高效液相串联质谱法同时测定中成药中5-甲氧基补骨脂素、8-甲氧基补骨脂素和欧前胡素

Simultaneous determination of 5-methoxypsoralen,8-methoxypsoralen and imperatorin in duliang dropping pills by ultra performance liquid chromatography tandem mass spectrometry

建立了同时测定中成药都梁滴丸中5-甲氧基补骨脂素、8-甲氧基补骨脂素和欧前胡素的超高效液相-串联质谱检测方法。都梁滴丸以甲醇超声提取45 min,提取液稀释后经Waters Oasis HLB固相萃取小柱净化,采用WatersACQUITY UPLC BEHC18（50 mm×2.1 mm,1.7μm）色谱柱,以乙腈（A）和水（B）为流动相进行梯度洗脱。采用电喷雾电离正离子模式,多反应监测模式检测,以保留时间和离子比定性,外标法定量。方法的定量限8-甲氧基补骨脂素和5-甲氧基补骨脂素均为0.3 mg/kg;欧前胡素为0.75 mg/kg。3种化合物在25~500μg/L范围内均呈线性,相关系数r〉0.99。在高,中,低三种添加水平的平均回收率为82.0%~107.4%日内精密度（RSD%）为1.1%~12%。

An analytical method based on ultra performance liquid chromatography tandem mass spectrometry has been developed for the determination of 5-Methoxypsoralen,8-Methoxypsoralen and Imperatorin.Duliang Dropping pills were extracted supersonically with methanol for 45 min,the extract was cleaned up by Oasis HLB solid phase extraction cartridge.After the chromatographic separation was performed on a Waters ACQUITY UPLC BEH C18 column（50 mm × 2.1 mm,1.7 μm）,qualitative and quantitative analysis was carried out on ESI-MS/MS in the positive ion mode using multiple reaction monitoring.Identification was achieved by the retention time and the ion ratio,quantification was done by the external standard curve.The limits of quantification（LOQs） for 5-Methoxypsoralen,8-Methoxypsoralen and Imperatorin were 0.3mg/kg,0.3mg/kg and 0.75 mg/kg separately.All the linear ranges of three compounds were in the range of 25～500 μg/L,with correlations coefficient（r） more than 0.99.The mean recoveries at three spiked level were 82.0%～107.4%,with the relative standard deviations between 1.1% and 12%.The method is rapid,accurate,simple and adapts to the determination 5-Methoxypsoralen,8-Methoxypsoralen and Imperatorin in Chinese medicine.