Multiplexed separations for biomarker discovery in metabolomics: Elucidating adaptive responses to exercise training
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High efficiency separations are needed to enhance selectivity, mass spectral quality, and quantitative performance in metabolomic studies. However, low sample throughput and complicated data preprocessing remain major bottlenecks to biomarker discovery. We introduce an accelerated data workflow to identify plasma metabolite signatures of exercise responsiveness when using multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS). This multiplexed separation platform takes advantage of customizable serial injections to enhance sample throughput and data fidelity based on temporally resolved ion signals derived from seven different sample segments analyzed within a single run. MSI-CE-MS was applied to explore the adaptive metabolic responses of a cohort of overweight/obese women (BMI > 25, n = 9) performing a 6-wk high-intensity interval training intervention using a repeated measures/cross-over study design. Venous blood samples were collected from each subject at three time intervals (baseline, postexercise, recovery) in their naïve and trained states while completing standardized cycling trials at the same absolute workload. Complementary statistical methods were used to classify dynamic changes in plasma metabolism associated with strenuous exercise and training status. Positive adaptations to exercise were associated with training-induced upregulation in plasma l-carnitine at rest due to improved muscle oxidative capacity, and greater antioxidant capacity as reflected by lower circulating glutathionyl-l-cysteine mixed disulfide. Attenuation in plasma hypoxanthine and higher O-acetyl-l-carnitine levels postexercise also indicated lower energetic stress for trained women.
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