苦参药材生物碱成分的HPLC指纹图谱

HPLC fingerprint of alkaloid components of Sophora flavescens Ait

目的:采用高效液相色谱法研究并建立苦参药材生物碱成分的HPLC指纹图谱,同时进行定性和定量相似度评价。方法:采用Lichrospher ODS-2色谱柱(250mm×4.6mm,5μm),甲醇-乙腈(1∶1)为有机相,0.1%氨水为水相,梯度洗脱,流速1.0mL/min,检测波长210nm,测定苦参药材的HPLC指纹图谱,统计分析确定共有模式;以最接近共有模式的苦参药材为特征对照,采用夹角余弦法(Cosθ)和相对欧氏距离法(RED)对不同产地苦参药材的HPLC指纹图谱进行定性与定量相似度评价。结果:苦参高效液相色谱指纹图谱中,以氧化苦参碱为参照峰,确定了10个特征峰。相似度比较结果显示,不同产地药材样品夹角余弦都大于0.9,表明各产地苦参药材化学组成一致性较好,相对欧氏距离由0.1至0.4不等,可见药材中特征成分含量上存在差异。结论:该方法重复性好,可为科学评价及有效控制苦参药材的质量提供参考。

Aim： To establish a sensitive and specific HPLC method for the systematic evaluation of qualitative and quantitative similarities of alkaloid components of Sophora flavescens Ait. from different origins. Methods： Alkaloid components of Sophora flavescens were separated on a Lichrospher ODS-2 （ 250 mm×4.6mm,5μm） column with a linear gradient elution model using a mobile phase consisting of methanol-acetonitrile （1： 1） and 0. 1% aqueous ammonia at a flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm and the column temperature 30℃. The similarities of the corresponding HPLC fingerprints were evaluated by cosine （ Cos O） of vectors and relative Euclidean distance methods, in terms of the chemical constituents and the con- tents, respectively. Results： Ten common peaks were confirmed in the HPLC fingerprints of Sophora flavescens using oxymatrine as the reference peak. Similarity analyses indicated that no significant difference between differ- ent origins was found in the qualitative similarity although obvious quantitative similarity differences were observed. Conclusion： HPLC fingerprints show good repeatability. It can be used as a means to assess and control the quality of Sophora flavescens.