Discovery of Urinary Biomarkers of Seaweed Intake Using Untargeted LC–MS Metabolomics in a Three-Way Cross-Over Human Study

Seaweeds are a marine source rich in potentially bioactive components, and therefore have attracted attention since the middle of the twentieth century. Accurate and objective assessment of the intake of seaweeds to study their health effects is hampered by a lack of validated intake biomarkers. In this three-armed, randomized, cross-over study, an untargeted metabolomics approach was applied for discovering novel intake biomarkers. Twenty healthy participants (9 men and 11 women) were provided each of three test meals in a randomized order: 5 g of Laminaria digitate (LD), 5 g of Undaria pinnatifida (UP), or a control meal with energy-adjusted pea protein. Four urine samples and a 24 h pooled urine were collected along with blood samples at seven time-points. All samples were profiled by LC-ESI-QTOF-MS and the data were analyzed by univariate analysis and excretion kinetics to select putative intake biomarkers. In total, four intake biomarkers were selected from urine samples. They were identified as hydroxyl-dihydrocoumarin at Level III, loliolid glucuronide at level I, and isololiolid glucuronide at level II, while the last one remains unknown. Further identification and validation of these biomarkers by a cross-sectional study is essential to assess their specificity and robustness.