基于^(1)H-NMR代谢组学技术考察五味子鲜果采后“杀青”的代谢轮廓变化

Investigation of Metabolic Profile Changes of Fresh Fruit of Schisandrae Chinensis Fructus After Green Removing Based on ^(1)H-NMR Metabonomics

目的:采用核磁共振氢谱(^(1)H-NMR)的代谢组学技术寻找五味子鲜果采后用不同方式干制后的差异代谢物,探讨“杀青”可能对其代谢轮廓造成的影响。方法:以15份五味子鲜果为样品,每3份为1组,分别采用烘干、晒干、微波、烫漂、蒸制5种方法进行初加工。将得到的五味子样品采用50%甲醇提取并进行^(1)H-NMR分析,结合多元统计分析,比较各“杀青”样品与烘干、晒干样品间代谢物轮廓的差异。结果:通过^(1)H-NMR图谱结合Chenomx数据库共指认出27个化学成分,主要为氨基酸、生物碱、糖类及有机酸,这些代谢产物主要参与果实采后生理的能量代谢。多元统计分析结果显示,各组样品组间差异明显。分别以烘干样品和晒干样品为对照,通过变量投影重要性(VIP)值和S-plot筛选出17个差异代谢物(天冬酰胺、柠檬酸、葡萄糖、蔗糖、甘油磷酰胆碱、葫芦巴碱、丙氨酸、赖氨酸、甘油、亮氨酸、异亮氨酸、缬氨酸、尿苷、苏氨酸、蛋氨酸、苯丙氨酸和果糖);组间比较采用单因素方差分析,结果发现与烘干和晒干样品比较,经“杀青”处理的五味子样品中蔗糖和甘油磷酰胆碱的含量均明显升高(P<0.05,P<0.01),缬氨酸、亮氨酸、异亮氨酸、苏氨酸、苯丙氨酸和蛋氨酸的含量均显著降低(P<0.01),微波“杀青”处理的五味子中天冬酰胺的含量明显升高(P<0.05,P<0.01),而烫漂处理的五味子中葫芦巴碱和尿苷的含量均显著降低(P<0.01)。结论:五味子鲜果采后“杀青”后可能通过干扰其能量代谢活动从而抑制有效成分的消耗、分解和(或)转化。

Objective:To find the different metabolites of Schisandrae Chinensis Fructus fresh fruits dried in different ways by proton nuclear magnetic resonance(^(1)H-NMR)metabonomics technology,and to explore the possible effect of green removing on its metabolic profile.Method:Fifteen samples of Schisandrae Chinensis Fructus fresh fruits were used,and each 3 samples was divided into one group.The samples were processed by five methods,including drying,drying in the sun,microwave,blanching and steaming.These samples were extracted with 50%methanol and analyzed by ^(1)H-NMR.Combined with multivariate statistical analysis,the differences of the metabolic profile between green removing samples and dried samples,sun-dried samples were compared.Result:A total of 27 chemical components were identified by ^(1)H-NMR and Chenomx database,mainly including amino acids,alkaloids,carbohydrates and organic acids.These metabolites were mainly involved in the energy metabolism of fruit postharvest physiology.The results of multivariate statistical analysis showed that there were significant differences among the 5 groups.The dried and sun-dried samples were used as the controls,17 differential metabolites(asparagine,citric acid,glucose,sucrose,choline glycerophosphate,trigonelline,alanine,lysine,glycerol,leucine,isoleucine,valine,uridine,threonine,methionine,phenylalanine and fructose)were screened by variable importance in the projection(VIP)value and S-plot.One-way ANOVA was used for comparison between groups,the results showed that compared with dried and sun-dried samples,the contents of sucrose and choline glycerophosphate increased significantly(P<0.05,P<0.01)in samples treated with green removing,but the contents of valine,leucine,isoleucine,threonine,phenylalanine and methionine decreased significantly(P<0.01),the content of asparagine increased significantly(P<0.05,P<0.01)in samples treated by microwave,while the contents of trigonelline and uridine decreased significantly(P<0.01)in samples treated by blanching.Conclusion:The consumption,decomposition and/or transformation of active ingredients may be inhibited by interfering with its energy metabolism after the green removing of Schisandrae Chinensis Fructus fresh fruits.