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[摘要]
目的 建立同时测定不同产地溪黄草中绿原酸、隐绿原酸、咖啡酸和迷迭香酸的高效液相色谱法,比较评价不同产地溪黄草质量的差异性。方法 采用Cosmosil色谱柱(250 mm×4.6 mm,5μm),流动相甲醇–0.1%冰醋酸水溶液,梯度洗脱;体积流量1.0 mL/min;柱温30℃;检测波长330 nm;进样量10 μL。采集溪黄草中4个成分定量数据,运用SIMCA 13.0软件进行主成分分析(PCA),辅助结合正交偏最小二乘(OPLS)、聚类分析(HCA)进行判别。结果 绿原酸、隐绿原酸、咖啡酸和迷迭香酸分别在0.014~0.280、0.026~0.520、0.012~0.240、0.240~4.800μg线性关系良好;平均回收率分别为100.63%、99.08%、99.30%、101.23%,RSD值分别为0.92%、1.34%、2.05%、0.86%。经主成分分析得出,咖啡酸和迷迭香酸是两个不同产地溪黄草构成差异的主要原因。结论 HPLC法同时测定溪黄草中4种酚酸类成分绿原酸、隐绿原酸、咖啡酸和迷迭香酸较为稳定,其中咖啡酸、迷迭香酸可作为区分不同产地溪黄草的质量标准之一。
[Key word]
[Abstract]
Objective To establish an HPLC method for simultaneous determination of chlorogenic acid, cryptochlorogenic acid, caffeic acid, and rosmarinic acid in Isodon sorra (Maxim.) Kudo from different habitats, and to compare and evaluate the quality difference of I. sorra from different habitats. Methods The separation was carried out on Cosmosil column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of methanol-0.1% glacial acetic acid solution with gradient elution. The flow rate was 1.0 mL/min. The column temperature was set at 30℃, and the detective wavelength was set at 330 nm. The volume of injection was 10 µL. Quantitative data of 4 components in I. sorra were collected. PCA was carried out by SIMCA 13.0 Software, and the discrimination was assisted by OPLS and HCA. Results The linear ranges of chlorogenic acid, cryptochlorogenic acid, caffeic acid, and rosmarinic acid were 0.014-0.280, 0.026-0.520, 0.012-0.240, and 0.240-4.800 μg, respectively. The average recovery rates were 100.63%, 99.08%, 99.30%, and 101.23%, respectively. The RSD values were 0.92%, 1.34%, 2.05%, and 0.86%. The results of PCA showed that caffeic acid and rosmarinic acid were the main reasons for the differences in the composition of I. sorra from two different habitats. Conclusion Simultaneous determination of 4 phenolic acids in I. sorra by HPLC is relatively stable. Caffeic acid and rosmarinic acid can be used as one of the quality standards to distinguish I. sorra from different habitats.
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[基金项目]
国家重大研究发展计划项目(2017YFC1702906);江西省卫计委中医药科研项目(2018B144)