Ultra-PerformanceLiquid Chromatography–High-ResolutionMass Spectrometry and Direct Infusion–High-Resolution MassSpectrometry for Combined Exploratory and Targeted Metabolic Profilingof Human Urine

摘要

The application of metabolic phenotyping to epidemiological studies involving thousands of biofluid samples presents a challenge for the selection of analytical platforms that meet the requirements of high-throughput precision analysis and cost-effectiveness. Here direct infusion–nanoelectrospray (DI–nESI) was compared with an ultra-performance liquid chromatography (UPLC)–high-resolution mass spectrometry (HRMS) method for metabolic profiling of an exemplary set of 132 human urine samples from a large epidemiological cohort. Both methods were developed and optimized to allow the simultaneous collection of high-resolution urinary metabolic profiles and quantitative data for a selected panel of 35 metabolites. The total run time for measuring the sample set in both polarities by UPLC–HRMS was 5 days compared with 9 h by DI–nESI–HRMS. To compare the classification ability of the two MS methods, we performed exploratory analysis of the full-scan HRMS profiles to detect sex-related differences in biochemical composition. Although metabolite identification is less specific in DI–nESI–HRMS,the significant features responsible for discrimination between sexeswere mostly the same in both MS-based platforms. Using the quantitativedata, we showed that 10 metabolites have strong correlation (Pearson’s r > 0.9 and Passing–Bablok regression slope of 0.8–1.3)and good agreement assessed by Bland–Altman plots between UPLC–HRMSand DI–nESI–HRMS and thus can be measured using a cheaperand less sample- and time-consuming method. A further twentymetabolites showed acceptable correlation between the two methodswith only five metabolites showing weak correlation (Pearson’s r < 0.4) and poor agreement due to the overestimationof the results by DI–nESI–HRMS.