Abstract
NMR-based metabolomics investigations of human biofluids offer great potential to uncover new biomarkers. In contrast to protocols for sample collection and biobanking, procedures for sample preparation prior to NMR measurements are still heterogeneous, thus compromising the comparability of the resulting data. Herein, we present results of an investigation of the handling of cerebrospinal fluid (CSF) samples for NMR metabolomics research. Origins of commonly observed problems when conducting NMR experiments on this type of sample are addressed, and suitable experimental conditions in terms of sample preparation and pH control are discussed. Sample stability was assessed by monitoring the degradation of CSF samples by NMR, hereby identifying metabolite candidates, which are potentially affected by sample storage. A protocol was devised yielding consistent spectroscopic data as well as achieving overall sample stability for robust analysis. We present easy to adopt standard operating procedures with the aim to establish a shared sample handling strategy that facilitates and promotes inter-laboratory comparison, and the analysis of sample degradation provides new insights into sample stability.
Highlights
Cerebrospinal fluid (CSF), a secretion product of the central nervous system (CNS), is in dynamic exchange with the nerve tissue and circulates metabolites
We have investigated the reliability of a self-designed protocol for the preparation of CSF samples, which enables the acquisition of NMR data compatible with small molecules spectra libraries used for NMR metabolomic profiling, preventing data misinterpretation
Since CSF sample stability as well as interpretability of spectral data depend mainly on pH and, to a lesser extent, on osmolality and ionic strength, we have mainly focused on different aspects of pH control of CSF samples
Summary
Cerebrospinal fluid (CSF), a secretion product of the central nervous system (CNS), is in dynamic exchange with the nerve tissue and circulates metabolites. CSF metabolomics has received much attention in the research of neurological disorders like Alzheimer’s, Parkinson’s disease, multiple sclerosis or brain injury, amongst others [2,3,4,5,6,7,8]. NMR spectroscopic analysis of CSF in the field of metabolomics has gained importance due to the minimal requirement for sample preparation, and its ability to simultaneously detect and quantify a wide. A fairly simple composition, low lipid and protein content [9,10,11]—compared to biofluids like blood or urine—make CSF an ideal candidate for NMR analysis, and its feasibility was demonstrated some thirty years ago [12]. Handling of CSF samples still remains challenging [13] and some practical issues related to sampling procedure and sample stability, pH control [14], require special attention
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