HPLC-MS/MS法同时测定柴胡疏肝散水提物中12种主要成分的含量

Simultaneous determination of 12 main components in Chaihu Shugan San by HPLC-MS/MS

目的:建立高效液相色谱-串联质谱测定柴胡疏肝散水提物冻干粉中柴胡皂苷(A、B_(1)、C、D)、橙皮苷、柚皮苷、甘草苷、芍药苷、没食子酸、阿魏酸、槲皮素和异甘草素含量的方法。方法:采用Welch Ultimate XB-C_(18)色谱柱(2.1 mm×100 mm,5μm);流动相组成为A相:乙腈,B相:0.1%甲酸-水;以0.3 mL·min^(-1)的流速进行梯度洗脱,进样量为10μL;采用电喷雾离子源(ESI),负离子模式,以多反应监测方式(MRM)进行定量分析。结果:柴胡疏肝散水提物中的12种成分在检测范围内与峰面积之间的线性关系良好。测定结果表明,每克柴胡疏肝散水提物冻干粉中含:柴胡皂苷A 6.712 mg,柴胡皂苷B15.810 mg,柴胡皂苷C 16.700 mg,柴胡皂苷D 0.034 mg,橙皮苷101.600 mg,柚皮苷172.000 mg,甘草苷3.300 mg,芍药苷50.483 mg,没食子酸1.870 mg,阿魏酸4.945 mg,异甘草素0.032 mg,槲皮素0.051 mg。结论:本方法灵敏度高,简便可靠,可用于柴胡疏肝散水提物冻干粉中12种主要成分的定量分析,并为建立柴胡疏肝散的质量标准提供方法学基础。

OBJECTIVE To establish a high performance liquid chromatography-tandem mass spectrometry method for the determinations of saikosaponin(A,B_(1),C,D),hesperidin,naringin,liquiritin,paeoniflorin,gallic acid,ferulic acid,quercetin and isoliquiritin in freeze-dried powder of Chaihu Shugan San.METHODS A Welch Ultimate XB-C_(18)column(2.1 mm×100 mm,5μm)was employed.The mobile phase was composed of A:acetonitrile and B:0.1%formic acid-water.Gradient elution was performed at a flow rate of 0.3 mL·min^(-1)and an injection volume of 10μL.And electrospray ionization(ESI)was utilized for quantitative analysis by multiple reaction monitoring(MRM)in a negative ion mode.RESULTS Twelve components in water extract freeze-dried powder of Chaihu Shugan San had an excellent linear relationship with peak area within the detection range.The results showed that each gram of lyophilized powder of Chaihu Shugan San water extract contained:saikosaponin A 6.712 mg,saikosaponin B_(1)5.810 mg,saikosaponin C 16.700 mg,saikosaponin D 0.034 mg,hesperidin 101.600 mg,naringin 172.000 mg,liquiritin 3.300 mg,paeoniflorin 50.483 mg,gallic acid 1.870 mg,ferulic acid 4.945 mg,isoliquiritin 0.032 mg and quercetin 0.051 mg.CONCLUSION Sensitive,simple and reliable,this method may be used for the quantitative analysis of 12 main components in water extract of Chaihu Shugan San,and it provides methodological rationales for establishing the quality standard of Chaihu Shugan San.