HPLC-MS测定癃闭康泰片中贝母素甲、贝母素乙的含量

 目的建立癃闭康泰中贝母素甲、贝母素乙的高效液相色谱-质谱联用含量测定法。方法癃闭康泰片经碱化,用氯仿-甲醇(4∶1)提取生物碱有效部位,提取液浓缩,残渣用甲醇溶解后,进行高效液相色谱-质谱测定。色谱柱为Agilent Zorbax ex-tend C₁₈(2.1 mm×100 mm,5μm);流动相为乙腈和10 mmol·L^-1甲酸铵(pH 8.0),梯度洗脱(0～15 min:乙腈30%～55%;15～25min乙腈55%);流速0.2 mL·min^-1;柱温30℃。质谱条件为电喷雾电离源(ESI),以选择性正离子方式检测,贝母素甲和贝母素乙的选择性检测离子分别为m/z 432.4和m/z 430.4。结果贝母素甲和贝母素乙的的线性范围均为0.01～10 mg·L^-1,相关系数均大于0.999,检测限均为0.45μg·L^-1,方法回收率均大于95%,日内日间精密度(RSD)均小于5%。结论本方法专属性强,灵敏度高,线性关系良好,操作简便,适用于癃闭康泰中贝母素甲、贝母素乙的含量测定。

Simultaneous Determination of Peimine and Peiminine in Longbikangtai Tablet by Liquid Chromatogram-Mass Spectrometry

OBJECTIVE To develop a sensitive and specific LC-MS method for the determination of peimine and peiminine in Longbikangtain tablet.METHODS The drug was extracted with a mixed solution of chloroform-methanol(4∶1) after being alkalined.The LC-MS method was performed on a Agilent Zorbax Extend-C₁₈(2.1 mm×150 mm,5 μm).The mobile phase consisted of CH₃CN and 10 mmoL·L^-1 ammonium formate(pH 8.0) at a gradient elution.The flow rate was at 0.2 mL·min^-1.The temperature of column was 30 ℃.Esi was applied and operated in positive mode.The monitor ions were m/z 432.4(peimine) and m/z 430.4(peiminine).RESULTS The linear calibration curve of peimine and peiminine were observed in the concentration range of 0.01～10 mg·L^-1.The lower limit of quantification of peimine and peiminine were 0.45 μg·L^-1.The relative recoveries were more than 95%.The inter-and intra-day precision(RSDS) were below 5.0%. CONCLUSION The method is proved to be convenient,sensitive and rapid,and suitable for the determination of peimine and peiminine in Longbikangtain tablet.