A multi-analytical approach for metabolomic profiling of zebrafish (Danio rerio) livers

Abstract

A metabolomic study was performed to investigate the biochemical profiles of livers from male and female zebrafish (Danio rerio), using a multiple platform approach, incorporating ¹H NMR, GC/MS and LC/MS. The reproducibility and reliability of the three methods were validated prior to the assays. Major biomolecules detected using one method were also cross examined using the other techniques. These metabolites included carbohydrates, lipids, amino acids detected using ¹H NMR and GC/MS, and acetylcarnitine, choline and various phospholipids determined using ¹H NMR and LC/MS. Our findings suggest that ¹H NMR provided comprehensive information on glucose, amino acids, pyruvate and other smaller biochemical constituents of the zebrafish liver. On the other hand, GC/MS was able to assay cholesterol, saturated and unsaturated fatty acids, and LC/MS was ideal for the analysis of lipids/phospholipids. These techniques revealed that there are significant differences in the biochemical profiles of male and female zebrafish liver tissue extracts. Specifically, we noted that although there were no significant differences observed for the carbohydrate profile, the amino acid profile was rather different in male and female zebrafish liver. Furthermore, data from all three techniques revealed that although the saturated fatty acid profile was similar, the compositions of unsaturated fatty acids were different in the two phenotypes. The overall findings suggested that this multiplatform approach offers comprehensive coverage of a metabolome as well as provides valuable insight towards understanding the different biochemical profiles of a biosystem.