蒙药材砂引草HPLC指纹图谱及6个成分含量测定研究  

Study on HPLC fingerprint and determination of 6 components in Mongolian medicinal material Tournefortia sibirica Linnaeus

作　　者：郝俊生 包红英 斯日古冷 孟和 任娅婷 红艳[3] 辛颖[3,2] HAO Jun-sheng;BAO Hong-ying;Si-ri-gu-leng;MENG He;REN Ya-ting;HONG Yan;XIN Ying(Tongliao Market Inspection and Testing Center,Tongliao 028000,China;National and Local Joint Engineering Research Center for Mongolian Medicine Research and Development,Tongliao 028000,China;College of Mongolian Medicine,Inner Mongolia Minzu University,NMPA Key Laboratory for Quality Control of Traditional Chinese Medicine,Mongolian Medicine,Tongliao 028000,China;Zhalaite Banner Market Supervision Administration,Tongliao 028000,China)

机构地区：[1]通辽市市场检验检测中心,通辽028000 [2]蒙药研发国家地方联合工程研究中心,通辽028000 [3]内蒙古民族大学蒙医药学院,通辽028000 [4]扎赉特旗市场监督管理局,兴安盟137600

出　　处：《药物分析杂志》2024年第5期816-826,共11页Chinese Journal of Pharmaceutical Analysis

基　　金：内蒙古自治区重大基础研究开放课题(NMKJZX1702);内蒙古自治区卫生健康科技计划项目(202202259)。

摘　　要：目的:建立不同产地蒙药材砂引草Tournefortia sibirica Linnaeus的HPLC指纹图谱并测定其6个成分(咖啡酸、异槲皮苷、槲皮苷、迷迭香酸、紫草酸和丹酚酸B)的含量,为砂引草质量控制提供依据。方法:采用Agilent Eclipse Plus C_(18)色谱柱(250 mm×4.6 mm,5μm),以0.2%磷酸溶液(A)-乙腈(B)为流动相,进行梯度洗脱(0~60 min,9%B→38%B),检测波长330 nm。对不同产地砂引草进行分析,采用国家药典委员会“中药色谱指纹图谱相似度评价系统软件”(2012版),建立砂引草指纹图谱,同时测定6个成分含量。结果:HPLC指纹图谱共标定了不同产地砂引草中11个共有峰,指认了咖啡酸、异槲皮苷、槲皮苷、迷迭香酸、紫草酸和丹酚酸B 6个成分,17批砂引草药材与对照指纹图谱之间的相似度在0.931~0.996。经方法学考察,咖啡酸、异槲皮苷、槲皮苷、迷迭香酸、紫草酸和丹酚酸6个成分精密度试验RSD依次为1.9%、1.0%、1.4%、0.19%、1.1%和0.32%;重复性试验RSD依次为3.5%、2.3%、3.3%、0.14%、1.1%和0.19%;稳定性试验RSD依次为0.80%、1.1%、1.7%、0.52%、0.54%和0.78%。咖啡酸、异槲皮苷、槲皮苷、迷迭香酸、紫草酸和丹酚酸B在0.0033~0.0295、0.0119~0.1073、0.0148~0.1328、0.0928~0.8352、0.0689~0.1058、0.0688~0.6199μg·mL^(-1)浓度范围内线性关系良好(均为r=1.000);平均回收率在93.9%~102.3%,按干燥品计含量在0.004%~0.013%、0.030%~0.259%、0.032%~0.256%、0.256%~1.246%、0.018%~0.072%和0.062%~0.499%。结论:建立的砂引草HPLC指纹图谱及6个成分含量测定方法稳定、可靠、重复性好,可为砂引草质量控制及开发研究提供参考。Objective:To establish HPLC fingerprint and determine caffeic acid,isoquercitrin,quercitrin,rosmarinic acid,lithospermic acid and salvianolic acid B in Tournefortia sibirica Linnaeus,and to provide evidence for quality control of Tournefortia sibirica Linnaeus.Methods:The chromatographic separation was performed on an Agilent Eclipse Plus C_(18)column(250 mm×4.6 mm,5μm)with gradient elution(0-60 min,9%B→38%B)of 0.2%phosphoric acid(A)and acetonitrile(B).The detection wavelength was 330 nm.The column temperature was kept at 40℃and the flow rate was 1.0 mL·min^(-1).The HPLC fingerprint of all batches of Tournefortia sibirica Linnaeus was established using Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 edition)and the common peaks were identified by reference standards.Six constituents in Tournefortia sibirica Linnaeus were quantified.Results:Eleven common peaks were confirmed,and 6 common peaks were identified by reference standards including caffeic acid,isoquercetin,quercetin,rosmarinic acid,purple oxalic acid,and salvianolic acid B.The similarities of 17 batches samples were 0.931 to 0.996.By the methodology examination,RSDs for the precision test were 1.9%,1.0%,1.4%,0.19%,1.1%and 0.32%,respectively.RSDs for the reproducible test were 3.5%,2.3%,3.3%,0.14%,1.1%and 0.19%,respectively.RSDs for the stability test were 0.80%,1.1%,1.7%,0.52%,0.54%and 0.78%,respectively.Caffeic acid,isoquercitrin,quercitrin,rosmarinic acid,lithospermic acid and salvianolic acid B had good separation and showed good linearity in their respective linear ranges.The average recoveries ranged from 95%to 105%,and the contents(calculated with reference to the dried drug)were between 0.004%to 0.013%,0.030%to 0.259%,0.032%to 0.256%,0.256%to 1.246%,0.018%to 0.072%and 0.062%0.499%,respectively.Conclusion:The established HPLC fingerprint and quantification method is stable and reliable,which can provide basis for the quality control of Tournefortia sibirica Linnaeus.

关 键 词：蒙药材 砂引草 指纹图谱 聚类分析 主成分分析 含量测定 咖啡酸 异槲皮苷 槲皮苷 迷迭香酸 紫草酸 丹酚酸B 

分 类 号：R917[医药卫生—药物分析学]