[关键词]
[摘要]
目的 建立经典名方苓桂术甘汤HPLC指纹图谱及3种成分含量测定方法,为经典名方苓桂术甘汤物质基准的研究提供参考。方法 色谱柱为CNW Athena C18(250 mm×4.6 mm,5μm),乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱(0~10 min,5%~10% A;10~20 min,10%~15% A;20~50 min,15%~30% A;50~75 min,30%~40% A;75~95 min,40%~50% A;95~111 min,50%~95% A),体积流量为1.0 mL/min,进样量为10 μL,柱温30℃。指纹图谱及甘草苷、甘草酸铵含量测定的检测波长为230 nm,桂皮醛为290 nm。采用国家药典委员会出版的"中药色谱指纹图谱相似度评价系统"(2012年版),建立10批苓桂术甘汤指纹图谱,并使用HPLC法同时测定3种指标成分含量。结果 对10批苓桂术甘汤进行研究,其指纹图谱相似度均大于0.9,标定了24个共有峰,各峰分离度较好。含量测定结果表明其甘草苷、甘草酸铵含量较高。经方法学考察,3种成分在一定浓度范围内呈良好的线性关系;精密度RSD值均小于1.0%;甘草苷、桂皮醛、甘草酸铵的平均加样回收率分别为 98.35%、101.51%、102.59%,RSD均小于2.5%;样品在48 h内稳定;方法的重复性良好。结论 建立的经典名方苓桂术甘汤指纹图谱及3个成分的含量测定方法简便、稳定、准确可靠,可用于苓桂术甘汤物质基准的质量控制。
[Key word]
[Abstract]
Objective To establish an HPLC fingerprint of classical named Linggui Zhugan Decoction and determination method of the content of three compounds, and provide reference for the study of the material basis of the classical Linggui Zhugan Decoction. Methods The method was performed by high performance liquid chromatography with CNW Athena C18 (250 mm×4.6 mm, 5 μm) and gradient elution with acetonitrile (A)-0.1% phosphoric acid (B) as the mobile phase, gradient elution:0-10 min, 5%-10%; 10-20 min, 10%-15% A; 20-50 min, 15%-30% A; 50-75 min, 30%-40% A; 75-95 min, 40%-50% A; 95-111 min, 50%-95% A. The flow rate was 1.0 mL/min, the injection volume was 10 μL, and the column temperature is 30℃. The fingerprint and the detection wavelength of glycyrrhizin and ammonium glycyrrhizinate were 230 nm, and cinnamic aldehyde was 290 nm. The chromatographic fingerprint evaluation system published by the State Pharmacopoeia Commission (2012 Edition) was used to establish the fingerprint of 10 batches of Lingguizhugan decoction, and the content of three kinds of index components were determined simultaneously by the established HPLC method. Results The research on the 10 batches of Lingguizhugan soup showed that the fingerprint similarity was greater than 0.9, and 24 common peaks were calibrated, and the peak resolution was good. The content determination results showed that the content of glycyrrhizin and ammonium glycyrrhizinate was high. According to the methodological investigation, the three components showed a good linear relationship within a certain concentration range; The precision RSD values were all less than 1.0%; The average recovery rates of glycyrrhizin, cinnamaldehyde and ammonium glycyrrhizinate were 98.35%, 101.51%, and 102.59%, respectively. The RSD is less than 2.5%; The sample is stable within 48 h, and the method has good repeatability. Conclusion The fingerprint method and the determination method of three components in the traditional Chinese medicine classical prescription named Linggui Zhugan Decoction established in this study are simple, stable, accurate and reliable, and can be used for the quality control of the Linggui Zhugan Decoction.
[中图分类号]
[基金项目]
国家中医药管理局中药标准化项目(ZYBZH-C-JL-25,ZYBZH-C-GZ-10)