基于HPLC指纹图谱和含量测定的皂角刺配方颗粒质量控制研究

Study on quality control of Gleditsiae Spina Dispensing Granules based on HPLC fingerprint and content determination

目的:建立皂角刺配方颗粒的高效液相色谱(HPLC)指纹图谱及含量测定方法,为皂角刺配方颗粒的质量控制和评价提供参考依据。方法:采用资生堂MG C18色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.2%磷酸水溶液为流动相进行梯度洗脱,流速1.0 mL/min、检测波长260 nm、柱温35℃,建立皂角刺配方颗粒的HPLC指纹图谱及指标性成分原儿茶酸的含量测定方法。结果:皂角刺配方颗粒的HPLC指纹图谱均呈现了5个共有特征峰,并指认其中的2、4、5号峰分别为原儿茶酸、对羟基苯甲酸和香草酸。同一厂家的10批次皂角刺配方颗粒的相似度介于0.814~1.000之间,原儿茶酸含量为2.28~2.60 mg/g,批次间差异较小;不同厂家间配方颗粒差异较大,相似度介于0.580~0.956之间,原儿茶酸含量为0.26~2.60 mg/g,含量差异较大。结论:所建立的指纹图谱和含量测定方法简便、可行、重复性好,可用于皂角刺配方颗粒的质量评价和控制。

Objective:To establish high performance liquid chromatography(HPLC)fingerprint and content determination method of Gleditsiae Spina Dispensing Granules,and to provide reference for their quality control and evaluation.Methods:The HPLC fingerprint of Cleditsiae Spina Dispensing Granules was established and the content of the index component protocatechuic acid was determined by HPLC.The chromatographic separation was performed on a Shiseido MG Cig column(4.6 mm×250 mm,5μm)with acetonitrile-0.2%phosphoric acid aqueous solution as the mobile phase for a gradient elution.The detection wavelength was set at 260 nm with a flow rate of 1.0 mL/min and a column temperature at 35 C.Results:There were 5 common peaks in all fingerprint spectra of Cleditsiae Spina Dispensing Granules,and the peaks 2,4 and 5 were identified as protocatechuic acid,p-hydroxybenzoic acid and vanillic acid respectively.The similarity of 10 batches of Gleditsiae Spina Dispensing Granules from the same manufacturer was ranged from 0.814 to 1.000,with the contents of protocatechuic acid from 2.28 to 2.60 mg/g.However,the similarity of Gleditsiae Spina Dispensing Granules from different manufacturers varied greatly with the similarity ranged from 0.580 to 0.956,and the contents of protocatechuic acid from 0.26 to 2.60 mg/g.Conclusion:The established HPLC fingerprint and content determination method are simple,feasible and reproducible,and can be used for the quality evaluation and control of Gleditsiae Spina Dispensing Granules.