龙胆泻肝配方颗粒HPLC-DAD指纹图谱的建立及主要成分含量测定

Establishment of HPLC- DAD Fingerprints of Longdan Xiegan Formula Granules and Determination of Main Constituents

目的建立龙胆泻肝配方颗粒的HPLC-DAD指纹图谱,并测定配方颗粒中黄芩苷、龙胆苦苷和绿原酸的含量,为其质量控制提供依据。方法利用RP-HPLC-DAD法,采用Acclaim 120A C18色谱柱,在254nm处、40℃温度条件下,以乙腈-0.5%醋酸为流动相进行梯度洗脱。结果建立了龙胆泻肝配方颗粒的HPLC指纹图谱,以黄芩苷为参照,确定了10批样品的30个共有峰为特征峰,各色谱峰分离度良好、相似度较高(>0.95);黄芩苷、龙胆苦苷和绿原酸线性范围良好(r均大于0.996),精密度、稳定性、重复性良好(RSD均小于2%),加样回收率均大于97%,RSD<3%(n=5)。结论龙胆泻肝配方颗粒HPLC指纹图谱分析方法及黄芩苷、龙胆苦苷和绿原酸含量测定方法准确、简便、可行,均可作为龙胆泻肝配方颗粒的治疗控制标准。

Objective To establish an HPLC- DAD method for fingerprint analysis of Longdan Xiegan Formula Granules and to determine the contents of baicalin,gentiopicroside and chlorogenic acid to provide a scientific basis for the quality control. Methods Separation was performed using RP- HPLC- DAD and Acclaim 120 A C18 column. The gradient elution was performed by the mobile phase consisting of acetonitrile and 0. 5% vinegar acid aqueous with the detection wavelength of 254 nm,and the column temperature was set as 30℃. Results Thirty peaks were selected as the specific fingerprint peaks through 10 batches Longdan Xiegan Formula Granules with the identified of baicalin. Good similarities were obtained in the established fingerprint through similarity analysis and the resolution was good. The method showed good linearity of baicalin,gentiopicroside and chlorogenic acid with correlation coefficients not less than 0. 996. The RSD of precision repeatability and stability was no more than 2%,the sample recovery was between 97% and 101%,and the RSD was less than 3%（ n = 5）. Conclusion The method is accurate and simple,which can be used for quality control of Longdan Xiegan Formula Granules.