大叶蛇葡萄有效部位高效液相色谱指纹图谱研究

Analysis of Ampelopsis megalophylla Diels et.Gilg by HPLC Fingerprint

目的建立大叶蛇葡萄Ampelopsis megalophyllaDiels et.Gilg有效部位的高效液相色谱(HPLC)指纹图谱,以便完整地反映大叶蛇葡萄有效部位的内在化学信息,为科学评价及有效控制其质量提供可靠方法。方法利用HPLC-DAD方法,梯度洗脱,测定了10批大叶蛇葡萄药材有效部位的样品。色谱条件为:Agilent TC C18分析柱(250 mm×4.6 mm,5μm),柱温为30℃;流动相:乙腈-0.1%磷酸水,梯度洗脱,流速为1.0 ml/min;检测波长为:250 nm。结果建立了大叶蛇葡萄有效部位的HPLC指纹图谱,对10批不同采收时间的大叶蛇葡萄有效部位的样品进行了指纹图谱分析,标定了10个共有指纹峰,通过与对照品的保留时间及紫外光谱比较,1,5,6,9,10号峰分别鉴定为蛇葡萄素、花旗松素、槲皮苷、槲皮素、杨梅素。结论建立的HPLC指纹图谱能科学全面地反映大叶蛇葡萄的有效部位,可用于大叶蛇葡萄的质量控制。

Objective To establish an HPLC fingerprints of the effective part of Ampelopsis megalophylla Diels et. Gilg for reflecting the internal chemical information and evaluating its quality in a scientific and effective way. Methods HPLC method was applied for the quality assessment of Ampelopsis megalophylla Diels et. Gilg. The preparation of sample, the HPLC column, mobile phase, elution mode （isocratic and gradient） and gradient program were optimized in order to obtain high qualified HPLC profile. The HPLC system consisted of a Waters 600E pump and a 2996 photodiode -array detector（DAD）. Data were acquired and processed with the Empower workstation. HPLC analysis was performed on an Agilent TC C18 column （250 mm×4.6 mm,5 μm） with the mixture of acetonitrile and 0.1% phosphoric acid aqueous solution as the mobile phase in gradient mode. The column temperature was set at 30℃,the flow rate was 1.0 ml/min,and the detection wavelength was at 250 nm. Results The HPLC fingerprints of the effective part of Ampelopsis megalophylla Diels et. Gilg showed 10 common peaks in 10 batches of samples collected at different time. By comparison of the retention time and the on -line UV spectra of chemical standards,peak 1,5,6,9 and 10 were identified as ampelosin, taxifolin, quercitrin, quercetin and myricetin, respectively. Conclusion The HPLC fingerprint of the effective part of Ampelopsis. megalophylla Diels et. Gilg with high specificity can be used to control its quality.