MRS offers unique possibilities for non-invasive studies of biochemistry in the human brain in vivo. A growing body of evidence suggests that proton MRS may contribute to the clinical evaluation of a number of pathologies including ischaemia, tumours, epilepsy, metabolic and neuropaediatric disorders. In most cases results are expressed as ratios between metabolite signals obtained at certain experimental conditions. Presenting the results as metabolite signal ratios may lead to misinterpretation because such alterations can be due to changes in the content of either one of the metabolites or both, or may simply be due to changes in relaxation behaviour. Absolute quantitation of metabolite concentrations is therefore warranted. A number of studies using single volume proton MRS indicate that absolute quantitation of metabolite concentration is possible with respect to N-acetyl aspartate (NAA), total creatine, choline containing compounds, (Cho) and inositols (Ins). Internal standards (unsaturated water signal) as well as external standards have been used for signal calibration. Quality control with respect to signal linearity with concentration or with size of selected volume, selection efficiency, outer volume depression and signal contamination is essential for validation of the measurements. Furthermore, corrections for the influence of relaxation behavior are necessary. The results published so far indicate that the concentrations of NAA, total creatine, Cho and Ins in mmoles (kg wet weight)-1 range between 8.2 and 17.2 (mean 10.2), 5.9 and 11.6 (mean 7.2), 1.1 and 2.0 (mean 1.5) and 3.9 and 8.1 (mean 6.1), respectively. So far only a limited number of clinical studies has been published including studies of acute stroke, multiple sclerosis and Alzheimer's disease. The results are promising and encourage further exploitation of the utility of quantitative proton MRS in clinical practise.