Abstract
The stability of lipids and other metabolites in human body fluids ranges from very stable over several days to very unstable within minutes after sample collection. Since the high-resolution analytics of metabolomics and lipidomics approaches comprise all these compounds, the handling of body fluid samples, and thus the pre-analytical phase, is of utmost importance to obtain valid profiling data. This phase consists of two parts, sample collection in the hospital (“bedside”) and sample processing in the laboratory (“bench”). For sample quality, the apparently simple steps in the hospital are much more critical than the “bench” side handling, where (bio)analytical chemists focus on highly standardized processing for high-resolution analysis under well-controlled conditions. This review discusses the most critical pre-analytical steps for sample quality from patient preparation; collection of body fluids (blood, urine, cerebrospinal fluid) to sample handling, transport, and storage in freezers; and subsequent thawing using current literature, as well as own investigations and practical experiences in the hospital. Furthermore, it provides guidance for (bio)analytical chemists to detect and prevent potential pre-analytical pitfalls at the “bedside,” and how to assess the quality of already collected body fluid samples. A knowledge base is provided allowing one to decide whether or not the sample quality is acceptable for its intended use in distinct profiling approaches and to select the most suitable samples for high-resolution metabolomics and lipidomics investigations.Graphical abstract
Highlights
In targeted and non-targeted biomedical metabolomics and lipidomics studies, the quality of the achieved analytical data is dependent on the knowhow and experience ofanalytical chemists, but is highly dependent on the quality of the sample
If we are given a choice in the planning phase of a biomedical study, we suggest collecting plasma for mass spectrometry-driven metabolomics and lipidomics investigations [15]
S1P is a class of lipids well known to show alterations in their blood levels due to physiological conditions or diseases states [48–50], but Liu et al showed that alterations caused by these conditions are minor in comparison to changes based on pre-analytical errors [11], i.e., far below the S1P-d18:2 cutoff levels for the separation of plasma and serum samples of good and less suited quality for metabolomics and lipidomics studies
Summary
In targeted and non-targeted biomedical metabolomics and lipidomics studies, the quality of the achieved analytical data is dependent on the knowhow and experience of (bio)analytical chemists, but is highly dependent on the quality of the sample. - standardize collection procedure (e.g., tourniquet application time, site of venipuncture, collection order for different tubes, sample mixing, ...) - sample handling and transportation: standardize time period and temperature until centrifugation (continuous cooling is highly recommended) - processing delays: define acceptance criteria regarding sample stability - define centrifugation conditions for blood, urine, CSF (G force, temperature, time, brake use, etc.) - standardize the post-centrifugation period until storage or further processing (should be as short as possible, but fulfillable) - define volume and number of sample aliquots for long-term storage at −80 °C or below by the analytical needs - thaw samples at 4 °C and standardize accurate mixing of thawed samples (avoid repetitive freeze-thaw cycles and mark refrozen samples) - deviations from the protocol: specify recording and documentation (information should be accessible to all involved scientists) recommendations for solutions on how to get high-quality body fluids from the bedside to the bench for metabolomics and lipidomics analyses. No data exist to answer this question, but the tube for metabolomics and lipidomics should
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