天然产物研究与开发 ›› 2022, Vol. 34 ›› Issue (5): 824-835.doi: 10.16333/j.1001-6880.2022.5.011

• 开发研究 • 上一篇    下一篇

基于主成分分析的青熟酸枣果肉初生及次生代谢产物差异变化的研究

杨兴晶1,刘妍如1*,唐志书1*,宋忠兴1,常百金2,赵艳婷1,赵梦利1   

  1. 1陕西中医药大学 陕西省中药资源产业化协同创新中心 陕西省中药基础与新药研究重点实验室,咸阳 712083;2长春中医药大学,长春 130117
  • 出版日期:2022-05-28 发布日期:2022-05-23
  • 基金资助:
    财政部和农业农村部:国家现代农业产业技术体系资助项目( CARS-21);陕西省科技厅重点产业创新链(群)项目(2020ZDLSF05-08);陕西省高层次人才特殊支持计划&青年拔尖人才支持计划(陕组通字【2018】33号);咸阳市2020重大科技专项(2020K01-20)

Study on the differential changes of primary and secondary metabolites between green and ripe Zizyphus jujuba var. spinosa fruits based on principal component analysis

YANG Xing-jing1,LIU Yan-ru1 *,TANG Zhi-shu1 *,SONG Zhong-xing1,CHANG Bai-jin2,ZHAO Yan-ting1,ZHAO Meng-li1   

  1. 1Shaanxi Province Key Laboratory of New Drugs and Chinese Medicine Foundation Research,Shaanxi Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Shaanxi University of Chinese Medicine,Xianyang 712083,China;2Changchun University of Chinese medicine,Changchun 1301173,China
  • Online:2022-05-28 Published:2022-05-23

摘要:

本研究采用UPLC-Q-TOF-MS/MS技术对青酸枣果肉与熟酸枣果肉的主要成分类型进行分析,基于PCA主成分分析方法,分析两者初生/次生代谢物的差异,并对其生物合成途径变化进行分析。采用超高效液相色谱-四级杆-飞行时间质谱(UPLC-Q-TOF-MS/MS)法对青、熟酸枣果肉进行初生及次生代谢产物表征,初步鉴定出青、熟酸枣果肉中共有的成分有32种;其次以主成分分析(principal component analysis,PCA)、聚类分析及KEGG(Kyoto Encyclopedia of Genes and Genomes)通路富集分析进行差异初生代谢物筛选及相关代谢通路富集,将青、熟酸枣果肉样品主要归为2类且得到2个重要差异初生代谢物和3条重要代谢途径;然后,以质谱多反应检测(MRM)对筛选得到具有差异的初生代谢物和次生代谢物进行定量验证,对同一酸枣果肉中差异代谢物做定量方法学考察,其在0.002 3~800.00 μg/mL线性关系良好(r<0.999),方法重复性、供试品稳定性、仪器精密度均良好,平均加样回收率为9638%~104.15%,RSD为1.16%~2.46%(n = 6),最后进行生物合成途径的关联和阐释。本研究建立的方法能很好的说明青、熟酸枣果肉中的成分差异。分析结果显示,和风味相关的儿茶素及表儿茶素等黄酮类及其初生代谢产物有统计学差异,与其功效相关的三萜类成分中齐墩果酸与酸枣仁皂苷B有统计学差异。

关键词: 酸枣果肉, UPLC-Q-TOF-MS/MS, 初生代谢物, 次生代谢物, 主成分分析

Abstract:

In order to characterize the component differentiation from unripe and ripe Zizyphus jujuba var. spinosa,the approach of mass spectrometric technology was adopted to make the coupled chemometric analysis in this experiment.Firstly,the ultra performance liquid chromatography tandem time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) technology was utilized to identify the main components from the unripe and ripe fruits.In addition,32 of the primary and secondary metabolites from the two forms of fruits were characterized,and they were classified into 5 kinds of structures.Next,primary metabolites and secondary metabolites from these two forms of fruits were clustered and their varied components were derived via the principal component analysis (PCA).According to PCA analysis,the samples were divided into two categories and 10 of components were selected as the candidate variables to differentia those two forms of fruits.Then,the candidate components from the primary metabolites to secondary metabolites were enriched in their metabolite pathways and biosynthetic pathways via KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis.At the same time,the candidate components were determined by a mass spectrometry multiple reaction detection (MRM) based methodology.With a good linear relationship from 0.002 3 to 800.00 μg/mL (r<0.99),the average recoveries were 96.38%-104.15% with the RSDs of 1.16%-2.46% (n=6).According to the established methodology,the effects related components from two growing periods of Zizyphus jujuba var. spinosa were oleanolic acid and jujuboside B,which involved in triterpenoids pathways;the flavor-related components were catechin and epicatechin,which involved in flavonoids pathways.

Key words: Zizyphus jujuba var. spinosa , fruits, UPLC-Q-TOF-MS/MS, primary metabolites, secondary metabolites, PCA

中图分类号:  R284.1