苗药虎耳草的HPLC指纹图谱及5种成分的测定研究

Study on fingerprint of Saxifraga stolonifera herbs and determination of five components by HPLC

目的建立苗药虎耳草药材的HPLC指纹图谱及5种成分定量测定方法,为虎耳草质量控制提供参考。方法采用Agilent Eclipse XDB-C18(150 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.1%磷酸水溶液(B)为流动相,进行梯度洗脱;体积流量1.0 m L/min,检测波长254 nm,柱温35℃。对21批不同产地的虎耳草建立HPLC指纹图谱,采用国家药典委员会"中药色谱指纹图谱相似度评价系统(2004A)"对结果进行分析处理,并选取主要成分建立多指标测定方法。结果指纹图谱研究所建立的方法结果良好。21批不同产地虎耳草的HPLC指纹图谱中,确定11个共有峰,并指认了其中5个色谱峰,分别是原儿茶酸、没食子酸、岩白菜素、槲皮素-5-O-β-D-葡萄糖苷和槲皮苷,21批虎耳草药材与对照药材的相似度差异较大。将5个指认成分作为主成分建立多指标测定方法,其线性关系良好,范围分别为0.052 8~0.844 8、0.020 96~0.335 36、0.241 6~3.865 6、0.130 8~2.092 8和0.023 68~0.378 88μg;5种成分的回收率分别为96.64%、100.72%、96.62%、103.71%、96.75%,RSD均小于2%。21批药材中5种成分的质量分数分别在0.07~0.40、0.19~4.36、1.42~5.98、0.42~6.86、0.11~1.51 mg/g。结论所建立的方法简便可靠、耐用性好,可作为虎耳草药材质量控制方法。

Objective To establish a method to research the fingerprint and determine the ingredients of Saxifraga stolonifera by HPLC, which aimed at providing reference for the quality control of Saxifraga stolonifera. Methods Agilent Eclipse XDB-C18（150 mm × 4.6 mm, 5 μm） column was used as the stationary phase, and the mobile phase consisted of acetonitrile（A）-0.1% phosphoric acid with gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was 254 nm, and the column temperature was maintained at 35 ℃. The result would be analyzed by SOP of Similarity evaluation system for chromatographic fingerprint of TCM. Results The results obtained by the method of fingerprint analysis are good. In the fingerprints, 11 peaks were selected as the common peaks to evaluate the similarities among 21 batches of S. stolonifera collected from different regions, and five contents of them were indentified as protocatechuic acid, gallic acid, bergenin, quercetin-5-O-β-D-glucoside, and quercitrin. The similarities between standard herb and each determined herb showed a lot of differences from others. The determination method showed that there weregood linear relationships of five figured contents in the range of 0.052 8—0.844 8, 0.020 96—0.335 36, 0.241 6—3.865 6, 0.130 8—2.092 8, and 0.023 68—0.378 88 μg. Moreover, the recoveries rates of five figured contents are 96.64%, 100.72%, 96.62%, 103.71%, 96.75%,and all RSDs were lower than 2%. The contents of 5 components in the gathered herbs were determined by external standard method in the range of 0.07—0.40, 0.19—4.36, 1.42—5.98, 0.42—6.86, and 0.11—1.51 mg/g. Conclusion The method established in this study is simple, reliable and durable, which can provide a scientific basis for the quality control of S. stolonifera.