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Quantitative nuclear magnetic resonance (NMR) spectroscopy of blood plasma is widely used to investigate perturbed metabolic processes in human diseases The reliability of biochemical data derived from these measurements is dependent on the quality of the sample collection and exact preparation and analysis protocols Here, we describe systematically, the impact of variations in sample collection and preparation on information recovery from quantitative proton (1H) NMR spectroscopy of human blood plasma and serum The effects of variation of blood collection tube sizes and preservatives, successive freeze-thaw cycles, sample storage at -80degreesC, and short-term storage at 4 and 20degreesC on the quantitative lipoprotein and metabolite patterns were investigated Storage of plasma samples at 4degreesC for up to 48 h, freezing at -80degreesC and blood sample collection tube choice have few and minor effects on quantitative lipoprotein profiles, and even storage at 4degreesC for up to 168 h caused little information loss In contrast, the impact of heat-treatment (56degreesC for 30 min), which has been used for inactivation of SARS-CoV-2 and other viruses, that may be required prior to analytical measurements in low level biosecurity facilities induced marked changes in both lipoprotein and low molecular weight metabolite profiles It was conclusively demonstrated that this heat inactivation procedure degrades lipoproteins and changes metabolic information in complex ways Plasma from control individuals and SARS-CoV-2 infected patients are differentially altered resulting in the creation of artifactual pseudo-biomarkers and destruction of real biomarkers to the extent that data from heat-treated samples are largely uninterpretable We also present several simple blood sample handling recommendations for optimal NMR-based biomarker discovery investigations in SARS CoV-2 studies and general clinical biomarker research
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Journal_of_Proteome_Research
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Quantitative in-vitro diagnostic nmr spectroscopy for lipoprotein and metabolite measurements in plasma and serum: recommendations for analytical artifact minimization with special reference to COVID-19/SARS-CoV-2 samples. (Special Issue: Proteomics and its application in pandemic diseases.)
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