User‐friendly relative quantification procedure for gas chromatography/mass spectrometry–based plasma metabolome analysis

Abstract

RationaleRecently, metabolome analysis has been applied to a variety of research fields, but differences between batches or facilities can cause discrepancies in the results of such analyses. To resolve these issues using comprehensive metabolome analysis, in which it is difficult to perform quantitative analyses of all detected metabolites, internal standard compounds are used to obtain relative metabolite levels. This study investigated gas chromatography/mass spectrometry–based plasma metabolome analysis methods that are superior to relative quantification using internal standard compounds.MethodsIn experiment I, four analyses were performed under different analytical conditions at one facility, and then the data from the four analyses were compared. In experiment II, the same samples were analyzed at three facilities, and then the data from the three facilities were compared.ResultsRegarding the relative values obtained through comparisons with the internal standard compound, differences in the analytical results were observed among the four analytical conditions in experiment I and among the three facilities in experiment II, and the differences observed among the three facilities (experiment II) were larger. When correction was performed using plasma as a quality control, which is the procedure suggested in this study, these differences were markedly ameliorated.ConclusionThe suggested procedure involves the analysis of a plasma standard as a quality control for each batch and the calculation of relative target plasma to quality‐control plasma values for each metabolite. This is an easy and low‐cost method and could be readily employed by researchers during comprehensive plasma metabolome analysis.