HPLC-DAD-ELSD法同时测定清热八味胶囊中8个成分的含量及主成分分析

Simultaneous Determination of 8 Components in Qingre Bawei Capsules by HPLC-DAD-ELSD and Principal Component Analysis

目的:建立高效液相色谱-二极管阵列检测-蒸发光散射检测(HPLC-DAD-ELSD)法同时测定清热八味胶囊中胆红素、羟基红花黄色素A、胡黄连苷Ⅱ、胡黄连苷Ⅰ、胆酸、肉桂酸、紫堇灵、山柰素的含量,并运用主成分分析进行评价。方法:以L9(34)正交试验优化提取条件。采用Kromasil C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%甲酸溶液为流动相,梯度洗脱;流速为1.0 ml·min^(-1),检测波长为280,367,403 nm;ELSD检测器漂移管温度为85℃,氮气流速为2.5 L·min^(-1)。运用SPSS 22.0软件对8个成分含量结果进行主成分分析。结果:8个成分在各自范围内线性关系良好(r>0.99);平均加样回收率为97.3%~104.0%,RSD在0.36%~1.23%之间(n=9);清热八味胶囊最佳提取方法为30 ml 50%甲醇超声(250 W,40 k Hz)提取60 min。主成分分析结果显示,8个有效成分中胆红素、胡黄连苷Ⅱ、肉桂酸、山柰素和紫堇灵可反映清热八味胶囊质量效应成分大部分信息。结论:本方法准确度、灵敏度高、精密度及重复性好,结合主成分分析,可为进一步完善清热八味胶囊质量控制提供参考。

Objective:To establish an HPLC-PAD-ELSD method for the simultaneous determination of bilirubin,hydroxysafflor yellow A,picroside-Ⅱ,picroside-Ⅰ,cholic acid,cinnamic acid,corynoline and kaempferol in Qingre Bawei capsules,and to evaluate the drug quality with principal component analysis.Methods:The L_(9)(3^(4))orthogonal design was carried out in order to optimize the extraction method.The analysis was performed on a Kromasil C18column(4.6 mm×250 mm,5μm)with the mobile phase consisting of acetonitrile-0.1% formic acid(gradient elution)at the flow rate of 1.0 ml·min^(-1).The detection wavelength was set at 280,367 and 403 nm.The ELSD conditions were as follows:the temperature of the drift tube was maintained at 85℃ and the flow rate of nebulizing gas was 2.5 L·min^(-1).The results of the above eight components were analyzed by SPSS 22.0 software.Results:The 8 constituents showed good linear relationships within their own ranges(r>0.99),the average recoveries were 97.3%-104.0% with the RSDs of 0.36%-1.23%(n=9).The optimal extraction method of Qingre Bawei capsules was ultrasonic extraction(250 W,40 k Hz)with 30 ml methanol for 60 min.The principal components analysis showed that bilirubin,picroside-II,cinnamic acid,kaempferol and corynoline reflected the most quality information of Qingre Bawei capsules.Conclusion:The HPLC method has high accuracy,sensitivity,precision and repeatability.Combined with principal component analysis,it can provide reference for further improving the quality control of Qingre Bawei capsules.