American journal of physiology. Endocrinology and metabolism | 2021
High throughput LC-MS method to investigate postprandial lipemia: Considerations for future precision nutrition research.
Abstract
Elevated postprandial lipemia is an independent risk factor for cardiovascular disease, yet methods to quantitate post-meal handling of dietary lipids in humans are limited. This study tested a new method to track dietary lipid appearance using a stable isotope tracer (2H11-oleate) in liquid meals containing three levels of fat (low-LF, 15g; moderate-MF, 30g; high-HF, 60g). Meals were fed to 12 healthy men (mean±SD, age 31.3±9.2y, BMI 24.5±1.9 kg/m2) during four randomized study visits; the HF meal was administered twice for reproducibility. Blood was collected over 8h postprandially, TG-rich lipoproteins (TRL) and particles with a Svedberg flotation rate >400 (Sf>400, n=8) were isolated by ultracentrifugation, and labeling of two TG species (54:3 and 52:2) quantified by LC-MS. Total plasma TRL-TG concentrations were three-fold greater than Sf>400-TG. Both Sf>400- and TRL-TG 54:3 were present at higher concentrations than 52:2 and singly-labeled TG concentrations were higher than doubly-labeled. Further, TG 54:3 and the singly-labeled molecules demonstrated higher plasma absolute entry rates differing significantly across fat levels within a single TG species (P<0.01). Calculation of fractional entry showed no significant differences in label handling supporting the utility of either TG species for appearance rate calculations. These data demonstrate the utility of labeling research meals with stable isotopes to investigate human postprandial lipemia while simultaneously highlighting the importance of examining individual responses. Meal type and timing, control of pre-study activities, and effects of sex on outcomes should match the research goals. The method, optimized here, will be beneficial to conduct basic science research in precision nutrition and clinical drug development.