毛冬青根和茎的HPLC指纹图谱研究

Study on the roots and stems fingerprints of Ilex pubescens by HPLC

目的建立毛冬青根与茎的HPLC指纹图谱测定方法,并对比商品毛冬青及其同属近缘植物的指纹图谱,为鉴定毛冬青根、茎以及近缘植物提供理论依据。方法采用Luna-C18(250 mm×4.6 mm,5μm)色谱柱,柱温为30℃,乙腈(10%异丙醇)(A)-0.4%甲酸水(B)梯度洗脱,流速1.0 mL·min-1,进样量20μL。ELSD气化温度40℃,氮气压力2.5 Bar,增益值为2。结果分别建立了毛冬青根与茎的HPLC-ELSD指纹图谱,并对商品进行相似度比较。毛冬青根中标示出15个共有峰,茎中标示出17个共有峰,并指认其中4个共有峰,分别为Ilexgein A、Ilexsaponin A1、Ilexsaponin B1、Ilexsaponin B2。结论所建立的HPLC指纹图谱可用于鉴别毛冬青根、茎及其常见近缘植物,可为毛冬青的质量控制提供有效手段。

OBJECTIVE To develop the HPLC fingerprint of the roots and stems fingerprints of Ilex pubescens and compare fingerprints similarity with commercial herbs and its related species. METHODS The conditions of experiment were confirmed and the validation of the method was also tested. The chromatography conditions were defined as Luna -C18 （250 mm ×4.6 mm,5μm） , and the temperature of column was 30℃. The mobile phase was acetonitrile containing 10% propan - 2 - o1（A） - 0.4% formic acid （B） with the program of gradient elution, and the flow speed was 1.0 mL. rain-1. The injection volume was 20 μL. The ELSD conditions were as follows ： the temperature of drift tube was 40℃ , the gas pressure was 2.5 Bar, and the value of gain was 2. RESULTS The HPLC - ELSD fingerprints of the roots and stems were established, and the similarity of commercial herbs were compared. The total 15 peaks were selected as the characteristic peaks of the roots of llex pubescens and 17 peaks were selected as the characteristic peaks of its stems. Furthermore,four peaks in HPLC -ELSD chromatograms were assigned as Ilexgein A,Ilexsaponin A1, Ilexsaponin B1 and Ilexsaponin B2. CONCLUSION The characteristics of the roots and stems of llex pubescens has been obtained. The method can be used for the quality control of flex pubescens.