黄芪-当归药对配伍前后HPLC指纹图谱及主要化学成分变化

Study on HPLC Fingerprints and Main Chemical Composition Changes Before and After Compatibility of Astragali Radix and Angelicae Sinensis Radix

关键词:  黄芪  当归  配伍  高效液相色谱法  指纹图谱  成分变化

Key words:  Astragali Radix  Angelicae Sinensis Radix  compatibility  HPLC  fingerprint  component change

摘要:

目的 建立黄芪-当归（芪归）药对配伍前后HPLC指纹图谱及多指标成分同时测定的方法，探讨芪归药对配伍的物质基础。方法 采用水煎煮-喷雾干燥法制备芪归药对及黄芪、当归提取物；采用Shim-Pack VP-ODS（250 mm×4.6 mm，5 μm）色谱柱，柱温30℃，以水-甲醇溶液为流动相进行梯度洗脱，流速1.0 mL·min^-1，检测波长280 nm，建立芪归药对配伍前后HPLC指纹图谱及毛蕊异黄酮、毛蕊异黄酮葡萄糖苷、芒柄花素、阿魏酸、Z-藁本内酯等多指标成分同时测定的方法；求取指纹图谱共有模式，标定共有峰并分析相对峰面积变化，进行色谱峰归属，分析药对配伍前后指标性成分含量变化。结果 所建HPLC指纹图谱及多指标成分同时测定方法精密度良好，溶液中各待测成分在3 d内稳定，各色谱峰RSD值均<3.0%，5种指标成分在各自线性范围内均有良好的线性关系，回收率均符合相关规定，各样品指纹图谱相似度均>0.9；芪归药对配伍后共标定出26个共有峰，其中9个来自黄芪，6个来自当归，7个同时来自黄芪和当归，4个色谱峰是新生成物质；芪归药对配伍共煎后，黄芪中毛蕊异黄酮及芒柄花素含量均显著降低（P<0.05），当归中阿魏酸及Z-藁本内酯含量均显著降低（P<0.01）。结论 芪归药对配伍后HPLC指纹图谱变化显著，总体表现为多数化学成分含量减少、个别成分含量增加并有新化合物生成。

Abstract:

OBJECTIVE To establish methods for HPLC fingerprints and simultaneous determination of multi-index components before and after compatibility of Astragali Radix and Angelicae Sinensis Radix, and to explore material basis of their compatibility. METHODS The extracts of Astragali Radix, Angelicae Sinensis Radix and their compatibility were prepared by water decoction-spray drying method. The separation was performed on a Shim-Pack VP-ODS column(250 mm×4.6 mm, 5 μm) with a gradient elution at a temperature of 30℃, with the mobile phase comprising of water-methanol flowing at 1.0 mL·min^-1 in a gradient elution manner, and the detection wavelength was set at 280 nm. The methods for HPLC fingerprints and simultaneous determination of multi-index components of calycosin, calycosin-7-glucoside, fermononetin, ferulic acid and Z-ligustilide before and after compatibility of Astragali Radix and Angelicae Sinensis Radix were established. The common patterns of fingerprints were obtained, the common peaks were calibrated, the relative peak area changes were analyzed, the peaks were assigned, and the content changes of multi-index components before and after compatibility were analyzed. RESULTS The established methods for HPLC fingerprints and simultaneous determination of multi-index components had good precision, the components in solution were stable within 3 d, and the RSD values of each chromatographic peak were <3.0%. The five index components showed good linear relationships within their own ranges, and the recovery rates were in compliance with relevant regulations. The fingerprint similarities of each sample were >0.9. After compatibility of Astragali Radix and Angelicae Sinensis Radix, a total of twenty-six common peaks were calibrated, nine of which were from Astragali Radix, six from Angelicae Sinensis Radix, seven from both of them, and four were newly formed. After compatibility, the contents of calycosin and fermononetin from Astragali Radix were significantly decreased(P<0.05), and the contents of ferulic acid and Z-ligustilide were significantly decreased in Angelicae Sinensis Radix(P<0.01). CONCLUSION After compatibility of Astragali Radix and Angelicae Sinensis Radix, HPLC fingerprints changed significantly, which generally reflected in content decrease of most chemical components, increase of individual components, and formation of new components.