Metabolite Ratios as Quality Indicators for Pre-Analytical Variation in Serum and EDTA Plasma
Sven Heiling,
Nadine Knutti,
Franziska Scherr,
Jörg Geiger,
Juliane Weikert,
Michael Rose,
Roland Jahns,
Uta Ceglarek,
André Scherag,
Michael Kiehntopf
Affiliations
Sven Heiling
Institute of Clinical Chemistry and Laboratory Diagnostics and Integrated Biobank Jena (IBBJ), University Hospital Jena, Am Klinikum 1, 07747 Jena, Germany
Nadine Knutti
Institute of Clinical Chemistry and Laboratory Diagnostics and Integrated Biobank Jena (IBBJ), University Hospital Jena, Am Klinikum 1, 07747 Jena, Germany
Franziska Scherr
Institute of Clinical Chemistry and Laboratory Diagnostics and Integrated Biobank Jena (IBBJ), University Hospital Jena, Am Klinikum 1, 07747 Jena, Germany
Jörg Geiger
Interdisciplinary Bank of Biological Material and Data Würzburg (IBDW), Straubmühlweg 2a, Haus A9, 97078 Würzburg, Germany
Juliane Weikert
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, 04103 Leipzig, Germany
Michael Rose
Institute of Clinical Chemistry and Laboratory Diagnostics and Integrated Biobank Jena (IBBJ), University Hospital Jena, Am Klinikum 1, 07747 Jena, Germany
Roland Jahns
Interdisciplinary Bank of Biological Material and Data Würzburg (IBDW), Straubmühlweg 2a, Haus A9, 97078 Würzburg, Germany
Uta Ceglarek
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, 04103 Leipzig, Germany
André Scherag
Institute of Medical Statistics, Computer and Data Sciences, Jena University Hospital, Bachstrasse 18, 07743 Jena, Germany
Michael Kiehntopf
Institute of Clinical Chemistry and Laboratory Diagnostics and Integrated Biobank Jena (IBBJ), University Hospital Jena, Am Klinikum 1, 07747 Jena, Germany
In clinical diagnostics and research, blood samples are one of the most frequently used materials. Nevertheless, exploring the chemical composition of human plasma and serum is challenging due to the highly dynamic influence of pre-analytical variation. A prominent example is the variability in pre-centrifugation delay (time-to-centrifugation; TTC). Quality indicators (QI) reflecting sample TTC are of utmost importance in assessing sample history and resulting sample quality, which is essential for accurate diagnostics and conclusive, reproducible research. In the present study, we subjected human blood to varying TTCs at room temperature prior to processing for plasma or serum preparation. Potential sample QIs were identified by Ultra high pressure liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) based metabolite profiling in samples from healthy volunteers (n = 10). Selected QIs were validated by a targeted MS/MS approach in two independent sets of samples from patients (n = 40 and n = 70). In serum, the hypoxanthine/guanosine (HG) and hypoxanthine/inosine (HI) ratios demonstrated high diagnostic performance (Sensitivity/Specificity > 80%) for the discrimination of samples with a TTC > 1 h. We identified several eicosanoids, such as 12-HETE, 15-(S)-HETE, 8-(S)-HETE, 12-oxo-HETE, (±)13-HODE and 12-(S)-HEPE as QIs for a pre-centrifugation delay > 2 h. 12-HETE, 12-oxo-HETE, 8-(S)-HETE, and 12-(S)-HEPE, and the HI- and HG-ratios could be validated in patient samples.
