UPLC-Q-TOF-MS结合化学识别模式的皖贝母指纹图谱研究

Research on the fingerprint of Fritillaria anhuiensis Bulbus by UPLC-Q-TOF-MS technique with chemical recognition mode

目的建立皖贝母超高效液相色谱-飞行时间质谱联用技术(UPLC-Q-TOF-MS)的指纹图谱及化学计量分析模式,为其质量评价及标准制定提供参考。方法采用CORTECS C18色谱柱(4.6 mm×150 mm,2.7μm),流动相为乙腈-0.1%甲酸和10 mmol/L甲酸铵水溶液进行梯度洗脱,流速0.4 ml/min,进样量为2.0μl。采用中药指纹图谱相似度评价系统(2012年版)对9批皖贝母样品指纹图谱进行相似度评价,通过液相色谱-质谱技术进行定性分析,并结合聚类分析、主成分分析、正交最小偏二乘-判别分析进行整体质量评价。结果建立了不同批次皖贝母的指纹图谱,并确定了21个共有峰,初步指认出12个共有峰。聚类分析、主成分分析、正交最小偏二乘-判别分析可将9批皖贝母聚为3类。结论本研究所建立的皖贝母指纹图谱结合化学模式识别方法灵敏度高、专属性强,可较好体现皖贝母的整体性与差异性,从而为皖贝母的质量评价及标准制定提供参考依据。

Objective To establish the fingerprint and stoichiometric analysis mode of Ultra Performance Liquid Chromatography Tandem Quadrupole Time-of-Flight Mass Spectrometry(UPLC-Q-TOF-MS)of Fritillaria anhuiensis Bulbus,so as to provide reference for its quality evaluation and standard formulation.Methods By setting the CORTECS C18 column at 4.6 mm×150 mm,2.7μm with the mobile phases of acetonitrile-0.1%formic acid and 10 mmol/L aqueous ammonium formate for gradient elution at a flow rate of 0.4 ml/min and an injection volume of 2.0μl.The TCM fingerprint similarity evaluation system(2012 version)was used to evaluate 9 batches of Fritillaria anhuiensis Bulbus samples.By using Liquid Chromatography-Mass Spectrometry(LC-MS)technique to make quantity analysis and by combining cluster analysis,principal component analysis and orthogonal least squares-discriminant analysis to make overall quality evaluation.Results The fingerprint profiles of different batches of Fritillaria anhuiensis Bulbus were established and 21 common peaks were identified,and 12 of them were initially identified.Cluster analysis,principal component analysis and orthogonal least squares-discriminant analysis were used to cluster the nine batches of Fritillaria anhuiensis Bulbus into three categories.Conclusion The fingerprint established in this study combined with the chemical pattern recognition method are highly sensitive and specific,which could reflect the overall characteristics and differences of Fritillaria anhuiensis Bulbus,providing reference for the quality evaluation of Fritillaria anhuiensis Bulbus and standardization of it.