In the last decades, the prevalence of neurodegenerative disorders such as Parkinson’s disease, dementia and stroke has significantly increased. The lack of early-stage biomarkers hereby is a major reason for poor outcomes and limited treatment options. Metabolism alterations are a major driver and readout of neurological implications; however, it can be assessed only to a very minor extent by non-invasive imaging technologies. By contrast, typical diagnostic biofluids like serum or urine reflect metabolic changes on the systemic scale and are not specific to the brain. That’s why the analysis of cerebrospinal fluid (CSF) using metabolomic techniques offers a great perspective to better stratify patients with neurological implications. However, as metabolites can readily degrade and CSF collection in a clinical environment is challenging, several confounders have to be considered. Furthermore, no single technique can measure the full CSF metabolome. Hence in this work we used directly collected and quenched CSF for a multi-modal metabolomics profiling approach to define the ex vivo baseline of the human CSF metabolome by robust Nuclear Magnetic Resonance (NMR) spectroscopy and latest state of the art liquid chromatography (LC) ion mobility spectrometry mass spectrometry (IMS-MS).