Clomiphene, trimetazidine, and meldonium are three common clinical drugs, and long-term consumption will cause damage to human health. Currently, animal-derived food contamination is a major concern, and these compounds have a high probability of entering the human body and remaining there for a long time. In this study, a liquid chromatography-tandem triple quadrupole mass spectrometry method was developed to determine the residues of three metabolic regulators in food. With 0.1% formic acid water-acetonitrile solution (25:75, V/V), samples were extracted ultrasonically. After MCX SPE Column Cleanup, the Shiseido PC HILIC column was used for separation, with positive ion multiple reaction monitoring mode monitoring and isotope internal standard method used for quantifying. In this optimized condition, both clomiphene and trimetazidine had detection limits of 0.25 µg/kg and quantification limits of 0.5 µg/kg. Meldonium had a detection limit of 2.5 µg/kg and a quantification limit of 5.0 µg/kg. Recovery rates ranged from 80.1–119.9%, and precision RSD was 1.95–15.4% (n = 6). This method is suitable for detecting food containing clomiphene, trimetazidine, and mildronium.
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