摘要:
目的:采用超声辅助提取法,建立高效液相色谱法(HPLC)同时测定不同产地地参中白桦脂酸、齐墩果酸和熊果酸的含量。方法:在单因素实验基础上,选择乙醇溶液体积分数、超声功率和提取时间为实验因子,三萜酸含量为响应值优化地参三萜酸的提取条件;并通过筛选色谱柱类型、流动相pH、柱温和流速优化色谱条件;结合方法学考察方法的适用性。结果:所测白桦脂酸、齐墩果酸、熊果酸等3种三萜酸分别在2.558~51.150、2.480~49.590、2.480~49.590 μg/mL范围内与各自峰面积积分值呈良好的线性关系,r值都大于0.9999;检出限(limit of determination,LOD)为5~7 ng,定量限(limit of quantification,LOQ)为16~22 ng,精密度、重复性和稳定性良好;平均回收率为100.52%~103.77%,RSD为0.99%~2.42%。不同产地地参的三萜酸化合物HPLC图谱具有一致性,白桦脂酸、齐墩果酸、熊果酸的单个三萜酸化合物含量有一定差异,地参中三萜酸类化合物的种类受产地差异影响小。结论:本方法操作简便、灵敏度高、准确性好、实用可靠,可用于地参样品中三萜酸的定量分析和质量控制,本研究为完善地参药材现行质量标准和开发利用提供了实验数据。
Abstract:
Objective:The aim was to establish a high performance liquid chromatography(HPLC)method with ultrasonic extraction to determine the content of betulinic acid,oleanolic acid and ursolic acid in Lycopus lucidus Turcz. var. hirtus Regel from different places simultaneously. Methods:On the basis of single factor experiment,the volume fraction of ethanol,ultrasonic power and time were selected as experimental factors,and triterpenic acid content was taken as response value to optimize extraction conditions. The chromatographic conditions were optimized by screening column types,pH of mobile phase,column temperature and flow rate. The applicability of the method was examined by the methodology. Results:The tested triterpenic acid showed a good linearity with their respective peak areasin the range of 2.558~51.150,2.480~49.590,2.480~49.590 μg/mL,r≥0.9999,the limit of determination was 5~7 ng,and the limit of quantification was 16~22 ng. The precision,reproducibility and stability were good,the average recovery was in the range of 100.52%~103.77%,and the RSD ranged from 0.99% to 2.42%. The HPLC chromatograms of Lycopus lucidus Turcz. var. hirtus Regel from different regions were similar. The content of betulinic acid,oleanolic acid and ursolic acid was different,and the species of triterpenic acid in different habitats was similar. Conclusion:This method is simple,sensitive,accurate,practical and reliable. It can be used for quantitative analysis and quality control of triterpenic acid in Lycopus lucidus Turcz. var. hirtus Regel samples. It provides experimental data for improving the current quality standard,development and utilization of Lycopus lucidus Turcz. var. Hirtus Regel.
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