¹H NMR characterization of normal human cerebrospinal fluid and the detection of methylmalonic acid in a vitamin B₁₂ deficient patient

Abstract

1H NMR was used to analyze human cerebrospinal fluid (CSF) from a group of neurological disease controls and from a vitamin B12 deficient patient. The spectra were acquired at either 7.06 or 9.40 T at ambient temperature with CSF freeze dried and reconstituted in 2H2O. 3-Trimethylsilyl propionate was used as an internal chemical shift and concentration reference. All of the CSF samples showed peaks for lactate, L-alanine, acetate, glutamine, citrate, creatine/creatinine and sugar resonances. There was good agreement between the metabolite concentrations as determined by NMR with those obtained using conventional chemical methods. 1D and 2D 1H NMR techniques along with J-coupling and T1 analysis were used to confirm the peak assignments. Methylmalonic acid could be detected and quantitated (ca 150 microM) in the CSF from the vitamin B12 deficient patient.

Dimethyl sulfone in human cerebrospinal fluid and blood plasma confirmed by one‐dimensional 1H and two‐dimensional 1H‐13C NMR

“…Until now, DMSO 2 had not been described as a regular metabolite in CSF. Commodari et al 11 described a singlet resonance at 3.17 ppm at pH 9.6 in two out of eight CSF samples (peak Y) and Wevers et al 2,3 reported this singlet at 3.14 ppm (pH 2.50) in plasma and CSF spectra (peak u10 in plasma and peak x 14 in CSF). However, in none of these in vitro NMR studies could this singlet be definitively assigned to DMSO 2 .…”

NMR‐based metabolomic analysis of cerebrospinal fluid and serum in neurological diseases – a diagnostic tool?

“…The feasibility of CSF metabolite analysis, using NMR spectroscopy, was initially demonstrated over 20 years ago (3,4) and recognition that CSF metabolites were related to clinical conditions (5) paved the way for further research in this area (6)(7)(8). Of particular relevance to the clinical application of CSF metabolomics was the identification of disease-specific metabolites in MS, degenerative dementia, inflammatory spinal disease, vitamin B12 deficiency and in inborn errors of metabolism (9)(10)(11)(12). More recently, studies of CSF metabolite profiles have demonstrated the differentiation of schizophrenia from healthy controls (13) and separation of viral, tubercular and bacterial meningitis (14).…”

10.1007/s10809-008-2002-7

“…They also observed a change in the concentration of formate in the cerebrospinal fluid of patients with various types of multiple sclerosis [53]. Other studies of cerebrospinal fluid were also performed using 1 H NMR spectroscopy: drug overdosage [56], bacterial meningitis [57], diabetes [58], vitamin B 12 deficiency [59], and other pathologies were examined. Moreover, an unidentified peak with a chemical shift of 2.82 ppm appeared in the spectra of cerebrospinal fluid from these patients; it is likely that this was a signal due to an N-methyl compound.…”