Identification of Potential Urinary Biomarkers for Bell Pepper Intake by HPLC–HRMS-Based Metabolomics and Structure Elucidation by NMR

Dietary biomarkers show great promise for objectively assessing the food intake in humans. In this study, potential urinary biomarkers for red bell pepper intake were identified based on a dietary intervention study and a comprehensive metabolomics approach. Spot urine samples from 14 volunteers were collected in the two phases of the study (control phase: abstaining from any bell pepper/paprika products; case phase: consumption of a defined amount of fresh red bell pepper and abstaining from any further bell pepper/paprika products) and analyzed by high-performance liquid chromatography high-resolution mass spectrometry (HPLC-HRMS). Comparison of the obtained metabolomics data using statistical analysis revealed that the respective urine metabolomes differ significantly, which was attributable to the bell pepper intake. Some of the most discriminating metabolites were selected and isolated from human urine for unequivocal structure elucidation by nuclear magnetic resonance (NMR) spectroscopy. Herein, seven novel glucuronidated metabolites most likely derived from capsanthin and capsianosides were identified, implying their potential application as dietary biomarkers for the entire Capsicum genus.