基于广泛靶向代谢组学的石金钱龟龟甲不同熬制时间的水提物成分差异分析

Analysis on the difference of components of water extracts of tortoise shell of yellow pond turtle at different decoction time based on widely targeted metabolomics

目的分析石金钱龟龟甲在不同熬制时间差异水提物的变化情况。方法本研究选取9个样品,分为3组进行代谢研究;首先通过非靶向代谢组技术检测混样,鉴定出样本中的水提物,然后将这些水提物与自建数据库结合,并基于广泛靶向代谢组学(TM)技术进行代谢分析,采用基于液相色谱与四级杆飞行时间质谱联用技术(LC/QTOF-MS/MS)检测平台、自建数据库以及重复相关性评估、主成分分析、正交偏最小二乘法判别分析等多元统计分析相结合的手段研究样品间的代谢组差异。结果共鉴定出1248个水提物,共鉴定出1248个水提物,共有8大类(氨基酸及其代谢物,杂环化合物,脂肪酰类,甘油磷脂类,醛、酮、酯类,辅酶和维生素,色胺、胆碱、色素、其他);熬制终期差异水提物相对含量最为丰富。结论该研究从TM广靶代谢组学角度初步解释了龟甲3个不同熬制时间的代谢产物的差异性及特征化合物,鉴于石金钱龟的代谢产物与人们的健康和营养有着密切的关系,可为金钱龟龟甲深加工产品开发与应用提供参考信息和理论依据。

Objective To analyze the change situation of water extracts of tortoise shell of yellow pond turtle at different decoction time.Methods A total of 9 samples in this experiment were selected and divided into 3 groups for metabolic study.Firstly,the mixed samples were detected by non-targeted metabolomics technology,and the water extracts in the samples were identified.Secondly,these water extracts were combined with the self-built database,and thirdly,the metabolic analysis was carried out based on Targeted Metabolome(TM)widely targeted metabolomics technology.The integrated multivariate statistical analysis,such as the detection platform based on liquid chromatography quadrupole-time-of-flight mass spectrometry(LC/QTOF-MS/MS),the self-built database and repeated correlation evaluation,principal component analysis and orthogonal partial least squares discriminant analysis were used to study the metabolic differences between samples.Results A total of 1248 water extracts were identified,in 8 categories(amino acids and their metabolites,heteroeyclic compounds,fatty acyl,glycerol phospholipids,aldehydes,ketones,esters,coenzymes and vitamins,tryptamine,choline,pigments,etc.),among which 113 different water extracts were screened for A vs.B.85 different water extracts were screened by B vs.C,187 different water extracts were screened by A vs.C.The relative content of differential water extract was the most abundant in the final cooking stage.Conclusion In this study,the differences and characteristic compounds of the metabolites of tortoise shell at three different decoction time were preliminarily explained from the perspective of TM widely targeted metabolomics.In view of the close relationship between the metabolites of yellow pond turtle and people’s health and nutrition,it can provide reference information and theoretical basis for the development and application of deep-processed products of tortoise shell of yellow pond turtle.