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 C18 色谱柱(250 mm ×4.6 mm,5 μm),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 0.003 3～0.029 5、0.011 9～0.107 3、0.014 8～0.132 8、0.092 8～0.835 2、0.068 9～0.105 8、0.068 8～0.619 9μg·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.
Abstract
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 C18 column(250 mm ×4.6 mm,5 μm)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 Tourne-fortia 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 exam-ination,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 stabil-ity test were 0.80％,1.1％,1.7％,0.52％,0.54％and 0.78％,respectively.Caffeic acid,isoquercitrin,quer-citrin,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 quantifi-cation method is stable and reliable,which can provide basis for the quality control of Tournefortia sibirica Linnaeus.
基金项目
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(202202259)
NETL
NSTL
万方数据