多波长融合玄参饮片HPLC指纹图谱研究

Study on the Multi-wavelength HPLC Fingerprinting of Scrophulariae Radix

摘要:

目的：建立玄参多波长融合HPLC指纹图谱，为玄参的质量提供新方法。方法：采用多波长融合HPLC测定不同产地玄参饮片。色谱条件：Thermo C₁₈柱，乙腈-0.1%甲酸溶液为流动相梯度洗脱。多波长检测（0~5 min为230 nm，5~13 min为210 nm，13~23 min为320 nm，23~50 min为278 nm，流速1 mL·min^-1），柱温30 ℃。结果：建立了多波长融合玄参饮片HPLC指纹图谱共有模式，确认25个共有峰，标定其中6个峰，10批玄参样品相似度均>0.90。结论：建立的指纹图谱色谱峰数较多，色谱信息量大，适用于玄参饮片的质量评价。

Abstract:

Objective: Using the multi-wavelength high performance liquid chromatography(HPLC) for testing 10 Scrophulariae Radix samples from Zhejiang Panan, Hubei Enshi, ect. To provide a new method for scientific evaluation and effective control of its quality, and establish the multi-wavelength HPLC fingerprinting of Scrophulariae Radix from different places. Method: Chromatographic conditions:Thermo C₁₈ column, gradient elution was applied with acetonitrile and 0.1% methane acid as the mobile-phase. Ultraviolet (UV) was used combined with multiple wavelength to detect the chromatographic peaks that cannot be displayed in the same channel. The wavelength timetable was as follows, from 0 to 5 min the wavelength was 230 nm;from 5 to 13 min was 210 nm;from 13 to 23 min was 320 nm;from 23 to 50 min was 278 nm. The flow rate was 1 mL·min^-1 and the column temperature was maintained at 30 ℃. Result: A HPLC fingerprinting pattern was set up based on multi-wavelength combined with UV, which demarcated 25 commom peaks and confirmed 6 peaks. The index similarty of those Scrophulariae Radix fingerprinting which analysised through similarity evaluation system for chromatographic fingerprint of TCM (Version 2004 A) were all greater than 0.90. Conclusion: The approach of multi-wavelength HPLC will help to differentiate and evaluate the quality of Scrophulariae Radix. The method is highly sensitive, characteristic peaks of large quantity, chromatographic large amount of information that will help evaluate the quality of Scrophulariaceae Radix.