Multiple sclerosis (MS) lesions have been shown by conventional methods to have major alterations in water and myelin lipid contents. To characterize these abnormalities more efficiently, NMR spectroscopy was used to evaluate water content by measuring relaxation times at 0.5 and 2.0 T. Subsequently, cholesterol content was obtained by using 1H NMR spectroscopy, while 31P NMR spectroscopy allowed measurement of the four main phospholipids and the inorganic phosphate concentrations in normal and pathological cerebral white matter samples. The relaxation times were significantly prolonged in MS lesions relative to normal white matter. Measured in different sample types, T1 and T2 times increased with water content. Moreover, the T2 of normal-appearing white matter was considerably lengthened. Analysis of white matter lipid composition using this method gave accurate values, which showed a significant decrease in phospholipids and cholesterol content in MS samples.