A new food surveillance framework links chemical detection, dietary exposure, and urinary modeling, revealing how some everyday supplements may carry anti-doping risks even without intentional substance use.
Study: Proactive surveillance of foodborne bioactives by integrated dietary exposure and urinary excretion assessment. Image Credit: Arkhipenko Olga / Shutterstock
In a recent study published in the journal npj Science of Food, researchers developed a novel food safety and anti-doping surveillance and exposure-assessment framework that uses a Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) platform to screen 331 compounds across diverse matrices, thereby potentially identifying products that may contribute to the unintentional consumption of pharmacologically active substances (PAS) through food and supplements.
The study employed this novel framework to quantitatively validate 214 analytes and apply the method to 78 commercial products. Study findings demonstrated that certain dietary supplements could yield modeled urinary concentrations exceeding World Anti-Doping Agency (WADA) reporting thresholds, necessitating a proactive, integrated framework for chemical exposure assessment.
Pharmacologically Active Food Exposure Background
The paper describes three main routes through which pharmacologically active substances may enter the food supply: 1. Naturally occurring constituents, 2. Unintended environmental or industrial contamination, and 3. The illegal adulteration of dietary supplements.
This research has shown that compounds such as higenamine, a β2-agonist, occur naturally in botanical sources, e.g., lotus seeds, while synthetic growth promoters like clenbuterol may enter the supply via contaminated meat products.
Furthermore, parallel clinical research has established that chronic exposure to these substances is associated with cardiovascular dysfunction, neuropsychiatric symptoms, and endocrine disruption, highlighting the importance of their routine monitoring.
Historically, analytical methodologies have been limited by narrow target scopes or single-matrix applicability. Reviews on the topic emphasize that this fragmented approach fails to capture the cumulative risk posed by complex modern diets.
For populations subject to biological monitoring, such as elite athletes, the lack of robust surveillance increases the probability of Adverse Analytical Findings (AAF) resulting from inadvertent ingestion rather than intentional performance enhancement.
Consequently, public health agencies and regulatory frameworks, such as the World Anti-Doping Agency (WADA), emphasize the critical need for high-throughput, multiplexed detection strategies that link chemical presence to biological relevance.
LC-MS/MS Surveillance Study Design
The present study addresses this knowledge gap by developing a novel unified workflow using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) to separate complex mixtures and identify individual molecules based on their chromatographic behavior and mass-to-charge transition patterns. The study specifically utilized a Vanquish HPLC system coupled with a TSQ Altis triple quadrupole mass spectrometer (Thermo Fisher Scientific) to establish a rapid 10-minute analytical runtime.
The method underwent rigorous validation across three representative matrices: pork, animal-derived products, oats, plant-based solids, and beverages and liquids. To extract the target substances, the study used the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) method, a multi-step purification process that uses salts and solvents to remove fats and proteins that might interfere with the results.
Following method standardization, the platform was applied to a dataset of 78 commercial products purchased from local markets and online retailers. Specifically, the platform was used to identify the components of commercial products, supplemented with food consumption data from the Republic of Korea to estimate each component’s daily intake (EDI).
Finally, the study leveraged a pharmacokinetic model to predict whether these substances would appear in a person’s urine at levels high enough to trigger a reportable anti-doping finding.
Supplement Contamination and Doping Findings
LC-MS/MS assays revealed a widespread presence of pharmacologically active bioactives, including prohibited or monitored substances, in commercial food and supplement samples, some at surprisingly high concentrations. Most surprisingly, octopamine, a stimulant, was detected in a beetroot supplement at a concentration of 368,000 μg/kg. This concentration far exceeds the trace amounts typically found in natural plant sources, suggesting potential fortification or the use of extreme extracts in the specific supplement.
The study’s excretion models revealed that consuming the beetroot supplement resulted in a predicted octopamine urinary concentration of 2,944 ng/mL, nearly triple the WADA reporting threshold of 1,000 ng/mL.
Furthermore, the study detected olodaterol, a prohibited β2-agonist used in asthma medication, in a puncture plant (Tribulus terrestris) supplement. The simulated urine concentration was 0.46 ng/mL. Since WADA maintains a zero-tolerance policy for this substance, any detectable amount would constitute a violation.
Similarly, coclaurine, a structural relative of the prohibited stimulant higenamine that raises analytical concerns because of its structural similarity to higenamine, was found at 788,000 μg/kg in a "guduchi" herbal supplement.
Notably, statistical analysis revealed that the novel quantitative method was highly reliable, with all 214 fully validated analytes demonstrating strong linearity, a correlation coefficient R2 ≥ 0.99. Furthermore, the study revealed that while the EDI for the general population was mostly below toxicological safety margins, older adults showed higher exposure to certain herbal powders due to habitual consumption.
Food Safety and Anti-Doping Implications
The present study and its novel LC-MS/MS platform mark a significant leap in food safety by moving from reactive testing to proactive surveillance. The platform demonstrated that a single, rapid test with a 10-minute runtime can effectively screen for hundreds of potential risks across various food groups.
The findings underscore a critical need for tighter regulation and better labeling, especially for the supplement industry, where "natural" does not always mean "safe" or "legal" in a sporting context.
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