UHPLC-Q-TOF-MS/MS分析浙贝母不同部位化学成分

Analysis of Chemical Composition in Different Parts of Fritillaria Thunbergii Using UHPLC-QTOF-MS/MS

目的 建立UHPLC-Q-TOF-MS/MS方法分析浙贝母不同部位(花、叶、茎和鳞茎)化学成分,探讨浙贝母中不同部位化学成分的差异。方法 浙贝母样品提取后,以水(含有25 mmol·L-1的醋酸铵及25 mmol·L-1的氨水)-乙腈为流动相,经UPLC BEH Amide色谱柱(2.1 mm×100 mm,1.7μm)梯度洗脱分离,在电喷雾离子源正离子模式下进行分析,通过二级串联质谱分析,对采集的图谱进行峰匹配、峰对齐、滤噪处理等进行特征峰提取;用主成分分析和正交偏最小二乘法-判别分析进行数据分析。结果 共鉴定出113个化学成分。主成分分析结果显示浙贝母花、鳞茎、茎、叶明显分为4类,表明不同部位之间化学成分差异显著;正交偏最小二乘法-判别分析结果显示浙贝母花和茎、鳞茎、叶之间差异化合物分别为16,19,12个。其中,异戊烯基腺嘌呤核苷、脯氨酰天冬氨酸、反玉米素、乙胺嗪在浙贝母花中含量较高。结论 本研究建立了一种快速、准确、可靠的浙贝母不同部位的鉴别方法,为浙贝母资源的综合开发利用提供了科学依据。

OBJECTIVE To establish a UHPLC-Q-TOF-MS/MS method to analysis the metabolic profiles of flowers,leaves,stems and bulbs of Fritillaria thunbergii Miq.,and study the difference of chemical composition in different parts.METHODS Samples were extracted and the separation of the analytes was carried out on the UPLC BEH Amide column(2.1 mm×100 mm,1.7 μm) using the mobile phase of water(containing 25 mmol·L-1 of ammonium acetate-25 mmol·L-1of ammonia) and acetonitrile by gradient elution.The detection was performed in the electrospray ionization positive mode.Through the analysis of the multistage tandem mass spectrometry,the characteristic peaks were extracted with mass spectrometry data peak matching,peak alignment,noise filtering.Principal component analysis and orthogonal partial least squares discriminant analysis were used to analyze the data.RESULTS The 113 chemical constituents were identified.The results of principal component analysis showed that the flowers,bulbs,stems and leaves of Fritillaria thunbergii Miq.could be obviously divided into four categories,indicating that there were significant differences in chemical components from different parts.The orthogonal partial least squares discriminant analysis results showed that the differential compounds between the flowers and the stems,bulb,leaves of Fritillaria thunbergii Miq.were 16,19 and 12,respectively.Among them,the contents of isoamyl adenine nucleoside,prolyl aspartic acid,anti-zein and ethyl amine were higher in the flowers.CONCLUSION This study establish a fast,accurate,and reliable method for differentiating the different parts of Fritillaria thunbergii Miq..Moreover,this study provides a scientific basis for the comprehensive development and utilization of Fritillaria thunbergii Miq.