基于液相色谱-串联质谱结合多元统计分析盐胁迫下金银花代谢产物积累的变化

Changes of Metabolite Accumulation in Lonicerae Japonicae Flos under Salt Stress Based on Liquid Chromatography-Tandem Mass Spectrometry Combined with Multivariate Statistical Analysis

目的探究盐胁迫下金银花中代谢产物积累的变化。方法采用超快速液相色谱-串联三重四级杆飞行时间质谱(UFLC-Triple TOF-MS/MS)对金银花盐胁迫组与对照组样品进行分析,用主成分分析(PCA)、偏最小二乘法判别分析(PLS-DA)进行数据处理,找出潜在的差异代谢物;通过一级质谱精确质荷比和二级质谱碎片信息,结合质谱数据库搜索、标准品比对及相关文献进行差异成分鉴定。然后用超快速液相色谱-三重四级杆线性离子阱串联质谱(UFLC-QTRAP-MS/MS)对15个目标差异成分进行测定,并作灰色关联分析(GRA)。结果金银花盐胁迫组与对照组的样品得到有效区分,筛选并鉴定31个差异成分;15个目标差异成分呈现不同的变化规律,GRA分析初步表明适度的盐胁迫可促进金银花中药效成分的合成与积累。结论为盐胁迫下金银花的质量评价及提高金银花的药用价值提供了新思路。

OBJECTIVE In order to study the changes of metabolite accumulation in Lonicerae Japonicae Flos under salt stress.METHODS The samples of Lonicerae Japonicae Flos after salt stress and the control group were detected by UFLC-Triple TOF-MS/MS.Principal component analysis(PCA)and partial least square discriminant analysis(PLS-DA)were used to process data and screen the potential differential metabolites.The differential constituents were identified according to MS accurate mass and MS/MS spectrometry fragmentation information,combined with the mass spectrometry database search,comparison with reference standards and related literatures.Furthermore,the contents of 15 index differential constituents were simultaneously determined by UFLC-QTRAP-MS/MS,and the grey correlation analysis(GRA)was performed.RESULTS The samples from the control group and the salt stress group were effectively distinguished.31 different constituents were screened and identified,of which 15 were quantified and the variation were different.GRA analysis showed that moderate salt stress could promote the accumulation of bioactive constituents in Lonicerae Japonicae Flos.CONCLUSION This study provides new insights for the quality evaluation of Lonicerae Japonicae Flos under salt stress and the improvement of its medicinal value.