Evaluation of metabolic variation in normal rat strains from a statistical analysis of 1H NMR spectra of urine

Abstract

NMR spectroscopic and statistical methods have been applied to investigate the biochemical variations within and between two phenotypically normal rat strains. The 600 MHz 1H NMR spectra of urine were acquired as part of a series of drug toxicity studies from 450 control rat urine samples from each of two strains of laboratory rat (Han Wistar and Sprague Dawley). The spectra were data-reduced to 256 intensity descriptors over a range of δ 0.2–10.0. The spectral variation was analysed both within and between strains in terms of the mean, standard deviation, skewness and kurtosis of each descriptor. It is demonstrated that spectral intensities corresponding to a number of endogenous metabolites do not show Gaussian distributions and there is evidence for bimodality for some metabolites. Additionally, despite the visual similarity of the NMR spectra from the two strains of rat, the descriptor distributions and the statistics derived from them revealed differences in the metabolite profiles, which clearly distinguished the two populations. This work is of value in the determination of biochemical normality and variability, and thus can be used to investigate, and place confidence limits on the biochemical deviations, which arise as a consequence of genetic modification or pathophysiological events.