UPLC-MS/MS法测定半枝莲中4个二萜烟酸酯的含量

Simultaneous determination of four diterpenoid nicotinates in Scutellariae Barbatae Herba by UPLC-MS/MS

目的:建立同时测定半枝莲中半枝莲碱A、半枝莲碱B、半枝莲萜C和半枝莲萜F含量的UPLC-MS/MS分析方法。方法:采用ACQUITY UPLC?BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以0.1%甲酸水溶液(A)-0.1%甲酸乙腈(B)为流动相,梯度洗脱(0~7 min,48%B→100%B;7~8 min,100%B;8~8.5 min,100%B→48%B;8.5~10 min,48%B),流速0.2 mL·min^(-1),柱温30℃;质谱采用电喷雾离子源(ESI),多反应监测(MRM)正离子模式进行扫描。结果:半枝莲碱A、半枝莲碱B、半枝莲萜C、半枝莲萜F质量浓度分别在0.3344~13.38、0.2231~8.923、0.1624~6.496、0.1988~7.954μg·mL^(-1)的范围内线性关系良好(r皆大于0.999);平均回收率(n=9)分别为98.2%、98.6%、99.9%、93.3%,RSD分别为4.2%、6.2%、4.6%、6.3%;27批样品中上述4个成分含量范围分别为15.44~168.4、14.98~118.23、13.31~62.40、12.37~117.67μg·g^(-1)。结论:所建立的方法可用于半枝莲中主要二萜烟酸酯类成分的定量分析。

Objective:To establish a quantitative method for the determination of scutebarbatine A,scutebarbatine B,scutebata C and scutebata F in Scutellariae Barbatae Herba by UPLC-MS/MS.Methods:The determination was performed on an ACQUITY UPLC?BEH C18(2.1 mm×100 mm,1.7μm)at 30℃.The mobile phase of 0.1%formic acid solution(A)-0.1%formic acid acetonitrile(B)was subjected to gradient elution(0-7 min,48%B→100%B;7-8 min,100%B;8-8.5 min,100%B→48%B;8.5-10 min,48%B)at the flow rate of 0.2 mL·min^(-1).The analytes were determined by electrospray ionization source(ESI)combined with multiple reaction monitoring(MRM)in positive ion mode.Results:Scutebarbatine A showed a good linearity in the range of 0.3344-13.38μg·mL^(-1)(r>0.999),scutebarbatine B showed a good linearity in the range of 0.2231-8.923μg·mL^(-1)(r>0.999),scutebata C showed a good linearity in the range of 0.1624-6.496μg·mL^(-1)(r>0.999),and scutebata F showed a good linearity in the range of 0.1988-7.954μg·mL^(-1)(r>0.999).The average recoveries(n=9)were 98.2%,98.6%,99.9%and 93.3%,and the corresponding RSDs were 4.2%,6.2%,4.6%and 6.3%,respectively.The contents of scutebarbatine A,scutebarbatine B,scutebata C and scutebata F in 27 batches of samples were 15.44-168.4,14.98-118.23,13.31-62.40,12.37-117.67μg·g^(-1),respectively.Conclusion:The method is competent in the quantification of the main diterpenoid nicotinates in Scutellariae Barbatae Herba.