HPLC波长切换法同时测定穴位敷贴方3种成分的含量  

Simultaneous determination of 3 components in Acupoint Application Prescription by HPLC wavelength switching method

作　　者：燕午宁 程丽芳 杨程[2] 李元圆[3] 何丽[1] 张鹏[2] 邹建新[2] 景晓平[1] YAN Wuning;CHENG Lifang;YANG Cheng;LI Yuanyuan;HE Li;ZHANG Peng;ZOU Jianxin;JING Xiaoping(Shanghai Children’s Hospital,Shanghai Jiao Tong University,Shanghai 200040,China;School of Materials and Engineering,Shanghai Jiao Tong University,Shanghai 200040,China;Hangzhou Medical College,Hangzhou 310053,Zhejiang,China)

机构地区：[1]上海市儿童医院,上海交通大学附属儿童医院,上海200040 [2]上海交通大学材料与工程学院,上海200040 [3]杭州医学院,浙江杭州310053

出　　处：《上海中医药大学学报》2020年第4期93-97,共5页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：上海市卫计委中医药科研专项研究项目(2016LP025);上海交通大学医工交叉基金项目(YG2015MS38,YG2016MS35)。

摘　　要：目的:建立高效液相色谱(HPLC)波长切换法同时测定穴位敷贴方中芥子碱硫氰酸盐、6-姜辣素、细辛脂素含量。方法:采用HPLC法检测穴位敷贴方中芥子碱硫氰酸盐、6-姜辣素、细辛脂素含量,并进行方法学考察。选用Agilent ZORBAX Extend-C18色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%磷酸水溶液(三乙胺调节pH 7.0)为流动相进行梯度洗脱,柱温40℃,流速1.0 ml/min,紫外检测器0~17 min检测波长为326 nm,17~50 min切换波长为280 nm,进样量10μl。结果:芥子碱硫氰酸盐在2.789~178.5μg/ml、6-姜辣素在2.508~160.5μg/ml、细辛脂素在1.134~72.60μg/ml范围内线性关系良好;重复性实验,芥子碱硫氰酸盐、6-姜辣素、细辛脂素的RSD分别为1.20%、1.02%、0.69%;精密度实验,三者的RSD分别为0.50%、0.93%、0.90%;稳定性实验,三者的RSD分别为1.23%、1.05%、1.26%;三者的平均加样回收率分别为100.63%、98.44%、98.87%,RSD分别为2.00%、1.20%、1.10%,均符合测定要求。测得不同配比的组方中3种成分的含量存在明显差异。结论:该方法精密度高,重复性和稳定性好,可用于穴位敷贴方中以上成分的含量测定。Objective:To establish a high performance liquid chromatography(HPLC)wavelength switching method for simultaneous determination of sinapine thiocyanate,6-gingerol and asarinin in Acupoint Application Prescription.Methods:The contents of sinapine thiocyanate,6-gingerol and asarinin in Acupoint Application Prescription were determined by HPLC,and the methodological investigation was carried out.The separation was performed on a Agilent ZORBAX Extend-C18 column(4.6 mm×250 mm,5μm)with acetonitrile-0.1%phosphoric acid containing water(triethylamine adjusted pH to 7.0)in gradient elution.The column temperature was 40℃,the flow rate was 1.0 ml/min,the detection wavelength of ultraviolet detector was 326 nm within 0-17 min and 280 nm within 17-50 min,and the injection volume was 10μl.Results:The linear ranges were 2.789-178.5μg/ml for sinapine thiocyanate,2.508-160.5μg/ml for 6-gingerol and 1.134-72.60μg/ml for asarinin.In replication test,the relative standard deviations(RSDs)of sinapine thiocyanate,6-gingerol and asarinin were 1.20%,1.02%and 0.69%respectively;in precision test,the RSDs were 0.50%,0.93%and 0.90%respectively;in stability test,the RSDs were 1.23%,1.05%and 1.26%respectively;the average recoveries of the three components were 100.63%,98.44%,98.87%,and the RSDs were 2.00%,1.20%and 1.10%,respectively.All data were conformed to the standard of determination.The contents of the three components were significantly different in prescriptions with different proportions.Conclusion:The method is repeatable and stable,with high precision,which could be used for content determination of above components in Acupoint Application Prescription.

关 键 词：穴位敷贴方 芥子碱硫氰酸盐 6-姜辣素 细辛脂素 高效液相色谱法 

分 类 号：R286.0[医药卫生—中药学]