Unstable tissue components (metabolites) are not easily captured and evaluated by traditional metabolomics methods. In this study, a comprehensive investigation of various sampling methods and storage conditions on the metabolomic profile of fish muscle was performed based on in vivo and ex vivo sampling. The GlobalStd algorithm and structure/reaction directed analysis under a linear mixed model were used to investigate the distinctive influences of these factors on the metabolomic profiles of fish tissue obtained via untargeted LC-MS analysis. Immediate analysis of samples yielded different metabolomic profiles compared to that of stored samples. Storage time was found to affect the metabolomic profile in a complex way, whereas storage temperature was shown to not substantially change this pattern. At the reaction level, metabolites involved in homologous series with butylation were shown stable during storage. Overall, our findings demonstrate that immediate instrumental analysis after in vivo solid phase microextraction (SPME) sampling and a reverse time series experimental design should be the preferred approaches for metabolomic profiling if unstable compounds are of interest.

中文翻译：

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鱼类肌肉组织的代谢特征随取样方法、储存策略和时间的变化

传统的代谢组学方法不容易捕获和评估不稳定的组织成分（代谢物）。在这项研究中，基于体内和离体采样，对各种采样方法和储存条件对鱼肌肉代谢组学的影响进行了全面调查。GlobalStd 算法和线性混合模型下的结构/反应定向分析用于研究这些因素对通过非靶向 LC-MS 分析获得的鱼组织代谢组学特征的独特影响。与储存的样品相比，样品的即时分析产生了不同的代谢组学特征。发现储存时间以复杂的方式影响代谢组学特征，而储存温度显示不会显着改变这种模式。在反应层面，与丁基化相关的同源系列的代谢物在储存期间显示稳定。总的来说，我们的研究结果表明，如果对不稳定化合物感兴趣，体内固相微萃取 (SPME) 采样和逆时间序列实验设计后的即时仪器分析应该是代谢组学分析的首选方法。