Abstract
BackgroundMetabolomics is a powerful emerging technology for studying the systems biology and chemistry of health and disease. Current targeted methods are often limited by the number of analytes that can be measured, and/or require multiple injections.MethodsWe developed a single-injection, targeted broad-spectrum plasma metabolomic method on a SCIEX Qtrap 5500 LC-ESI-MS/MS platform. Analytical validation was conducted for the reproducibility, linearity, carryover and blood collection tube effects. The method was also clinically validated for its potential utility in the diagnosis of chronic fatigue syndrome (CFS) using a cohort of 22 males CFS and 18 age- and sex-matched controls.ResultsOptimization of LC conditions and MS/MS parameters enabled the measurement of 610 key metabolites from 63 biochemical pathways and 95 stable isotope standards in a 45-minute HILIC method using a single injection without sacrificing sensitivity. The total imprecision (CVtotal) of peak area was 12% for both the control and CFS pools. The 8 metabolites selected in our previous study (PMID: 27573827) performed well in a clinical validation analysis even when the case and control samples were analyzed 1.5 years later on a different instrument by a different investigator, yielding a diagnostic accuracy of 95% (95% CI 85–100%) measured by the area under the ROC curve.ConclusionsA reliable and reproducible, broad-spectrum, targeted metabolomic method was developed, capable of measuring over 600 metabolites in plasma in a single injection. The method might be a useful tool in helping the diagnosis of CFS or other complex diseases.
Highlights
Metabolomics has recently emerged as a useful tool with broad applications in clinical and translational research such as disease biomarker discovery, clinical drug safety evaluation and precision medicine (Patti et al 2012; Becker et al 2012)
The 8 metabolites selected in our previous study (PMID: 27573827) performed well in a clinical validation analysis even when the case and control samples were analyzed 1.5 years later on a different instrument by a different investigator, yielding a diagnostic accuracy of 95% measured by the area under the receiver operator characteristic (ROC) curve
We described the full method with details on key analytical procedures such as method optimization, multiple reaction monitoring (MRM) transitions, linearity and carryover which would help other researchers in the field to adapt and apply this method to their own work
Summary
Metabolomics has recently emerged as a useful tool with broad applications in clinical and translational research such as disease biomarker discovery, clinical drug safety evaluation and precision medicine (Patti et al 2012; Becker et al 2012). It can be performed using either untargeted or targeted approaches. The 8 metabolites selected in our previous study (PMID: 27573827) performed well in a clinical validation analysis even when the case and control samples were analyzed 1.5 years later on a different instrument by a different investigator, yielding a diagnostic accuracy of 95% (95% CI 85–100%) measured by the area under the ROC curve. The method might be a useful tool in helping the diagnosis of CFS or other complex diseases
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