基于液相色谱-串联质谱技术的新疆一枝蒿植物代谢组学分析方法研究

Development of Plant Metabolomics Analytical Approach Based on Liquid Chromatography Tandem Mass Spectrometry in Artemisia rupestris L.

药用植物化学成分种类繁多、结构复杂,而传统方法难以获得全面反映其所含成分随时间、品种和生态环境等因素发生的变化信息,因此需要建立系统、全面的分析方法以进行整体物质基础研究。本研究开展了基于液相色谱-串联质谱(LC-MS/MS)技术的新疆一枝蒿植物代谢组学分析方法研究,并将其应用于该植物不同组织器官代谢组的差异分析。重点考察了样品前处理过程中的提取溶剂及比例、复溶溶剂比例、超声时间对非靶向代谢组学研究的影响,以代谢物覆盖度为主要指标选择了最优条件并进行了方法学验证。采用上述前处理方法和LC-MS/MS分析方法,获得了新疆一枝蒿根、茎、枝、叶、花不同组织器官的代谢组信息,并构建多变量统计模型进行分析,发现花与其它组织器官的代谢组存在显著差异。结合数据库检索与化合物质谱裂解规律分析,获得差异代谢物的结构类型,包括61个黄酮类、97个一枝蒿酮酸衍生物、7个绿原酸类化合物及15个其它类型化合物。进一步采用聚类热图分析针对上述180个差异代谢物在各组织器官的分布情况进行表征,初步揭示了各类化合物在不同组织器官的分布特征。本研究不仅为新疆一枝蒿植物化学成分的组织器官分布及其有效成分的合理利用提供了重要信息,同时为药用植物的质量控制、品种改良以及合理开发提供了新的研究策略。

Due to the diversity and complexity,the change of chemical components in medicinal plant according to the time,cultivated varieties or ecological condition is difficult to recognize using traditional phytochemistry method.In order to analyze the pharmacodynamics material basis in Uighur medicinal plant Artemisia rupestris L.in an effective and comprehensive way,a plant metabolomics approach was established based on liquid chromatography-tandem mass spectrometry(LC-MS/MS).This study firstly focused on the effect of extraction solvents,redissolve solvents and ultrasonic time on the untargeted metabolomics,then the optimal preparation condition was selected according to metabolites coverage.After methodology validation,the approach was applied to acquire metabolic information in root,stem,branchlet,leaf and flower of Artemisia rupestris L.The results showed that the metabolome in flower was obviously different with the other organs.Coupling with multivariate statistical analysis,a batch of differential metabolites were picked out,in which 61 flavonoids,97 rupestonic acid derivatives,7 chlorogenic acids and 15 other compounds were primarily recognized according to the characteristic fragmentation rules of specific structure type and database retrieval.Additionally,the distribution characteristics of the above 180 differential metabolites was illustrated by cluster heat map.In conclusion,this study provided important information about the rational utilization of effective parts from Artemisia rupestris L.,and offered a novel strategy for quality control,variety improvement and reasonable development of medicinal plants.