Determination of metabolite concentrations in human brain tumour biopsy samples using HR-MAS and ERETIC measurements

Accurate determination of the concentration of the metabolites contained in intact human biopsies of 10 glioblastoma multiforme samples was achieved using one-dimensional H-1 high-resolution magic angle spinning (HR-MAS) NMR combined with ERETIC (TM) (electronic reference to in vivo concentrations) measurements. The amount of sample used ranged from 6.8 to 12.9 mg. Metabolite concentrations were measured in each sample using two methods: with DSS (2,2-dimethyl-2-silapentane-5-sulfonate sodium salt) as an internal reference and with ERETICT (TM) as an external electronically generated reference. The ERETIC (TM) signal was shown to be highly reproducible and did not affect the spectral quality. The concentrations calculated by the ERETIC (TM) method in model solutions were shown to be independent of the salt concentration in the range typically found in biological samples (0-250mM). The EIRETIC (TM) method proved to be straightforward to use in tissues and much more robust than the internal standard method. The concentrations calculated using the internal DSS concentration were systematically found to be higher than those determined using the ERETIC (TM) technique. These results indicate a possible interaction of the DSS molecules with the biopsy sample. Moreover, variations in the sample preparation process, with possible loss of DSS solution, may hamper the quantification process, as happens in one of the ten samples analysed. In this study, the ERETIC (TM) method was validated on model solutions and used in brain tumour tissues. Calculated metabolite concentrations obtained with the ERETIC (TM) procedure matched the values determined in the same type of tumours by in vivo, ex vivo and in vitro methodologies. Copyright (C) 2008 John Wiley & Sons, Ltd.