不同产地野生与栽培伊贝母药材UPLC-ELSD指纹图谱研究

UPLC-ELSD Fingerprints of Fritillariae Pallidiflorae Bulbus of Wild Growing and Cultivated Species from Different Habitats

关键词:  伊贝母  野生  栽培  超高效液相色谱-蒸发光散射法  指纹图谱

Key words:  Fritillariae Pallidiflorae Bulbus  wild  cultivated  UPLC-ELSD  chromatographic fingerprint

摘要:

目的 建立伊贝母药材UPLC-ELSD指纹图谱，为有效控制其质量提供可靠的方法。方法 采用UPLC-ELSD方法，色谱柱为Waters Acquity UPLC^TM BEH C₁₈（100 mm×2.1 mm，1.7 μm），流动相为乙腈和0.02%三乙胺，梯度洗脱，流速0.25 mL·min^-1，柱温25℃，样品为室温度，ELSD漂移管温度40℃，喷雾器参数40%，增益值500，气体压力30psi。采用国家药典委员会颁布的中药色谱指纹图谱相似度评价系统2012 A版软件建立共有模式，以2种方法对29批野生与栽培伊贝母药材计算相似度评价图谱的相似性，同时利用聚类分析法分析结果。结果 29批伊贝母药材有16个共有特征峰，建立了UPLC-ELSD指纹图谱共有模式。各批次伊贝母药材相似度都≥0.801。29批野生与栽培伊贝母药材可通过系统聚类分成2~3类，同时定量测定了样品中的西贝母碱苷和西贝母碱。结论 所建立的UPLC-ELSD指纹图谱方法快速，可用于伊贝母药材的质量综合评价。

Abstract:

OBJECTIVE To establish a UPLC-ELSD fingerprint method for quality control of Fritillariae Pallidiflorae Bulbus. METHODS Waters Acquity UPLC^TM BEH C₁₈ chromatographic column (100 mm×2.1 mm, 1.7 μm) was adopted for gradient elute with the mobile phase consisting of acetonitrile-0.02% triethylamine-water. The flow rate was 0.25 mL·min^-1; the temperature of column was set at 25℃, sample temperature was room temperature and the Drift tube temperature was 40℃, spray parameter was 40%, the gain value was 500, the gas pressure was 30 psi. Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (version 2012 A) published by the State Pharmacopeia Committee of China was employed for the representative standard fingerprint, and the similarity of each chromatogram of Fritillariae Pallidiflorae Bulbus of wild and cultivated species from different habitats was also calculated by the two methods. Hierarchical cluster analysis was applied to investigating the test data. RESULTS The chromatographic fingerprint showed 16 characteristic peaks, which was established from 29 species of Fritillariae Pallidiflorae Bulbus and was generated as the representative standard fingerprint. The similarity of the chromatographic fingerprints from the 29 samples was ≥ 0.801. Twenty-nine samples were classified into 2-3 groups. And the contents of sipeimine-3β-D-glucoside and sipeimine were determined from 29 wild and cultivated species of Fritillariae Pallidiflorae Bulbus from different habitats. CONCLUSION The method of UPLC-ELSD fingerprint is time-saving and can be used for the comprehensive quality evaluation of Fritillariae Pallidiflorae Bulbus.