Unveiling the global landscape of anabolic steroid residues in beef via integrated targeted quantification and high-resolution suspect screening

Anabolic steroid (AS) residues in animal-derived foods continues to pose a significant concern due to their potential risks to food safety and human health. In this study, we developed an integrated strategy combining targeted quantification and suspect screening for the comprehensive detection of AS residues in beef. The analytical workflow included optimized solid-phase extraction followed by Ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) in scheduled Multiple Reaction Monitoring mode for targeted analysis, and UHPLC coupled with high resolution mass spectrometry (UHPLC-HRMS) in data-independent acquisition mode for suspect screening. Method validation confirmed satisfactory linearity (R² > 0.97), low detection limits (0.003-3 μg/kg), and acceptable recoveries (52.4-126.8 %). 15 AS compounds were identified across beef products from diverse global sources. The developed strategy enables sensitive and efficient detection of structurally diverse AS residues in complex food matrices, which provide robust technical support for ensuring food safety.