超高效液相色谱-串联质谱法分析贝母药材中5种生物碱

建立了贝母药材中贝母辛、西贝母碱、贝母素乙、贝母素甲和西贝母碱苷的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法。样品经碱化甲醇-三氯甲烷提取,用Acquity UPLC BEH C₁₈色谱柱分离,乙腈-0.1%甲酸溶液为流动相梯度洗脱,多反应监测(MRM)模式检测,外标法定量。结果表明,5种生物碱在0.000 5～0.05 mg/L范围内线性良好(r>0.999),在1.0、2.5、5.0 mg/kg加标水平下,回收率为78.6%～93.1%,相对标准偏差不大于8.6%。5种生物碱的检出限(LOD)为0.03 mg/kg,定量下限(LOQ)为0.1 mg/kg。该方法操作简便,灵敏度高,准确可靠,适用于贝母中上述5种生物碱的同时测定。

Determination of 5 Alkaloids in Fritillaria by Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry

A method of ultrahigh performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS ) was developed for the simultaneous determination of five alkaloids,including peimisine,sipeimine,peiminine,peimine and sipeimine-3 β-D -glucoside in Fritillaria.Samples were extracted with alkaline methanol-chloroform,and separated on an Acquity UPLC BEH C 18 column using acetonitrile solution containing 0.1 % formic acid as mobile phase by gradient elution.The target compounds were determined by UPLC-MS/MS under multiple reaction monitoring (MRM ) mode,and then quantified by external standard method.The calibration curves showed good linearities in the range of 0.000 5-0.05 mg/L with correlation coefficients ( r ) greater than 0.999.Recoveries for the five alkaloids at three spiked levels (1.0,2.5,5.0 mg/kg ) ranged from 78.6 % to 93.1 %,with relative standard deviations (RSD ) not more than 8.6 %.The limits of detection (LOD ) and limits of quantitation (LOQ ) were 0.03 mg/kg and 0.1 mg/kg,respectively.The established method was simple,sensitive,accurate and reliable,and could be applied in the simultaneous determination of the five alkaloids in Fritillaria.