Sample Pretreatment of Brain Tissues and Cerebrospinal Fluid for NMR Investigations

Samples of rat homogenated brain cortex and cerebrospinal fluid prepared by various protocols were investigated by high-resolution NMR. The stability of the major metabolites, biomarkers of neurode-generative diseases, was examined depending on the procedure of sample pretreatment. As shown, the optimal conditions of the sample pretreatment of brain tissues to NMR analysis involve mechanical homogenization followed by the addition of deuterated water and for prolong experiments, an extraction with a methanol-chloroform mixture (2: 1). Samples of cerebrospinal fluid for NMR study should be prepared in buffer media based on deuterated water to improve the stability of the preparation and prevent the drift of metabolite signals in the spectra of different samples during the experimentation. The chosen optimal conditions of sample pretreatment of brain tissue and cerebrospinal fluid can be used in future to study changes in the composition and concentration of metabolites in neurodegenerative diseases by virtue of NMR. The proposed methods allow more accurate the diagnosis of pathologies of the brain function at early stages.