HPLC法同时测定颠茄片5个指标成分含量及特征图谱分析

目的: 建立不同生产企业颠茄片中5个指标成分的含量和专属的颠茄片特征图谱检测方法.方法: 采用Welch XB-C₁₈(250 mm×4.6 mm,5 μm)色谱柱,流动相为甲醇-0.05%磷酸,梯度洗脱,流速1.0 mL·min^-1,检测波长344 nm柱温30 ℃.结果: 5个指标成分东莨菪苷、绿原酸、山柰素-3-O-半乳糖-(6→1)鼠李糖-7-O-葡萄糖苷、东莨菪内酯和芦丁在含量分析测定条件下,各组分分离,线性范围分别为32.60~978.0、10.00~300.0、31.90~957.0、36.3~1089和10.10~303.0 ng,加样回收率在97.6%~99.7%之间;在特征图谱研究中,以东莨菪内酯为参照峰,标定了16个共有峰,分析的13批颠茄片与共有模式之间有良好的相似性,相似度在0.9以上.结论: 建立的含量测定及特征图谱分析方法灵敏度高、结果准确、专属性强,可用于颠茄片的质量控制.

Objective: To establish a simultaneous determination of five main marker components and specific HPLC fingerprint method for the quality evaluation of belladonna tablets from different manufactories. Methods: A Welch XB-C₁₈ column(250 mm×4.6 mm, 5 μm)was adopted with methanol-0.05% phosphoric acid as the mobile phase for gradient elution at a flow rate of 1.0 mL·min^-1;the detection wavelength was 344 nm and the column temperature was 30 ℃. Results: The result showed that five main marker components(scopolin, chlorogenic acid, kaempferol-3-O-β-D(6-O-α-L-rhamnogalactoside)-7-O-β-D-glucoside, scopoletin and rutin)were separated well.The correlation coefficients were 0.9998, 0.9996, 0.9995, 0.9994 and, 0.9994 respectively, the linear ranges of five components were 32.60-978.0, 10.00-300.0 ng, 31.90-957.0, 36.30-1089 ng and 10.10-303.0 ng, respectively, and the average recoveries ranged from 97.6% to 99.72%. Totally 16 common peaks were calibrated with scopoletin as the reference peak.There were good similarities between the standard fingerprint chromatogram and each fingerprint chromatogram of the thirteen samples with their similarity coefficients no less than 0.9. Conclusion: The method is fast, accurate, sensitive and suitable for the quality control of belladonna tablets.