Abstract

Every day, analytical and bio-analytical chemists make sustained efforts to improve the sensitivity, specificity, robustness, and reproducibility of their methods. Especially in targeted and non-targeted profiling approaches, including metabolomics analysis, these objectives are not easy to achieve; however, robust and reproducible measurements and low coefficients of variation (CV) are crucial for successful metabolomics approaches. Nevertheless, all efforts from the analysts are in vain if the sample quality is poor, i.e. if preanalytical errors are made by the partner during sample collection. Preanalytical risks and errors are more common than expected, even when standard operating procedures (SOP) are used. This risk is particularly high in clinical studies, and poor sample quality may heavily bias the CV of the final analytical results, leading to disappointing outcomes of the study and consequently, although unjustified, to critical questions about the analytical performance of the approach from the partner who provided the samples. This review focuses on the preanalytical phase of liquid chromatography–mass spectrometry-driven metabolomics analysis of body fluids. Several important preanalytical factors that may seriously affect the profile of the investigated metabolome in body fluids, including factors before sample collection, blood drawing, subsequent handling of the whole blood (transportation), processing of plasma and serum, and inadequate conditions for sample storage, will be discussed. In addition, a detailed description of latent effects on the stability of the blood metabolome and a suggestion for a practical procedure to circumvent risks in the preanalytical phase will be given. Graphical The procedures and potential problems in preanalytical aspects of metabolomics studies using blood samples. Bias in the preanalytical phase may lead to unwanted results in the subsequential studies

Highlights

  • Metabolomics delineates biological phenotypes by profiling changes of endogenous metabolites [1]

  • All samples from the same individual clustered together regardless of the different conditions. These findings from a non-targeted metabolomics approach indicate that only minor changes may occur between freshly prepared plasma sample and whole blood placed on ice for up to 4 h [25]

  • The preanalytical challenges for metabolomics of blood are mainly caused by two factors: the stability of metabolites and the metabolic action of cells in the sample collection tube

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Summary

Introduction

Metabolomics delineates biological phenotypes by profiling changes of endogenous metabolites [1]. Several important preanalytical factors that may seriously affect the profile of the investigated metabolome in body fluids, including factors before sample collection, blood drawing, subsequent handling of the whole blood (transportation), processing of plasma and serum, and inadequate conditions for sample storage, will be discussed. This review will report critical aspects of the preanalytical phase for clinical metabolomics studies, with a special focus on the sampling of blood.

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