基于快速液相色谱-四级杆飞行时间串联质谱的浙贝母特征图谱研究

Fingerprint analysis of Fritillaria thunbergii using rapid resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry

该文建立了浙贝母甾体类生物碱RRLC-Q-TOF-MS特征图谱的质量评价方法。该方法采用Agilent Eclipse Plus C18色谱柱(2.1 mm×100 mm,1.8μm),流动相为乙腈-0.1%甲酸和10 mmol·L-1甲酸铵水溶液,梯度洗脱;检测器为Q-TOFMS并记录一级和二级质谱。在该色谱条件下,对12批浙贝母药材及其混淆品湖北贝母、伊犁贝母的RRLC-Q-TOF-MS特征图谱进行了测定和分析。结果从浙贝母、湖北贝母和伊犁贝母中鉴定了15个甾体类了生物碱,确定了浙贝母中9个共有特征峰。浙贝母和湖北贝母及伊犁贝母的特征图谱有明显差异。通过聚类分析和主成分分析,可对不同来源的浙贝母样品进行初步的质量评价。该文所建立的浙贝母甾体类生物碱成分的特征图谱特征性和专属性强,且方法快速、简便、可靠,可用于全面控制浙贝母的质量。

A fingerprint method for quality assessment of Fritillaria thunbergii was developed by rapid resolution liquid chro-matography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry （RRLC-Q-TOF-MS）. The separa-tion was performed using Agilent Eclipse Plus C18 column （2. lmm × 100 mm, 1.8 μm） by gradient elution with acetonitrile and 0. 1% formic acid aqueous solution （containing 10 mmol · L^-1 ammonium formate） as the mobile phase. Q-TOF-MS was used to obtain the accurate mass for precursor and product ions. Under this chromatographic and MS condition, 12 batches of F. thunbergii and its adul- terants （F. hupehensis and F. pallidiflora） were analyzed by RRLC-Q-TOF-MS. Fifteen steroidal alkaloids were identified from F. thunbergii, F. hupehensis and F. pallidiflora and nine were assigned as the common characteristic peaks for F. thunbergii. The RRLC-Q-TOF-MS fingerprint of F. thunbergii was different significantly with those of F. hupehensis and F. pallidiflora. The quality of 12 batches of F. thunbergii samples were finally evaluated by hierarchical clustering analysis （HCA） and principle component analysis （PCA）. This convenient and high specific method could be used to identify and evaluate the quality of the F. thunbergii.