Rapid Quantitative Platform of Extensive Bioactive Compounds in Foods for Exposure and Urinary Excretion-based Assessment

Abstract

Unintended dietary exposure to pharmacologically active substances from foods and supplements is an emerging challenge for food safety and anti-doping surveillance. These compounds—whether naturally occurring, introduced as contaminants, or present due to supplement adulteration—pose health risks and regulatory challenges. Here, we developed and validated a high-throughput liquid chromatography–tandem mass spectrometry (LC–MS/MS) method capable of simultaneously screening 332 compounds and quantitatively validating 214 analytes across diverse food matrices. Validation confirmed robust selectivity, sensitivity (LODs 0.00015–11.19 µg/kg), linearity (R² ≥ 0.99), accuracy (60.79–125.54%), and precision (CV ≤ 22.44%) in accordance with Codex Alimentarius guidelines. Applied to 78 commercial foods and supplements, the method revealed widespread occurrence of doping-related substances, including octopamine (368,000 µg/kg in beetroot supplement), olodaterol (9,170 µg/kg in Tribulus terrestris), and coclaurine (788,000 µg/kg in Tinospora cordifolia), along with caffeine, synephrine, and nicotine. Dietary exposure assessment identified coclaurine and synephrine as higher-risk contributors in older adults, while urinary excretion modeling showed supplement-derived octopamine and olodaterol could exceed World Anti-Doping Agency (WADA) reporting thresholds at recommended intake levels. This integrated framework, combining broad-spectrum LC–MS/MS analysis with dietary exposure and urinary excretion modeling, offers a scalable platform for proactive surveillance, regulatory oversight, and consumer protection in food safety and anti-doping contexts.