Tehran University of Medical Sciences Journal of Food Safety and Hygiene 2476-3241 12 2 2026 10 21 158 172 Serveh Anboyeh Department of Environmental Health Engineering, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Mojtaba Moazzen Department of Food Technology Research, National Nutrition and Food Technology Research Institute, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Nabi Shariatifar Department of Food Safety and Hygiene, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. 2026 08 31 2026 09 14 This research aimed to investigate methyltestosterone and androstenedione hormones in chicken meat using the quick, easy, cheap, effective, rugged, and safe (QuEChERS) technique and liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS). The optimization method was validated by assessing key parameters including recovery, limit of detection (LOD), limit of quantification (LOQ), and relative standard deviation (RSD). Recovery values varied from 76.3% to 116.7%. The LOD and LOQ for both hormones were determined to be 0.16 and 0.50 ng/g, respectively. Analysis of 72 chicken meat samples revealed that the levels of both hormones were below the LOD. Therefore, the concentrations of these compounds were found to be below established regulatory limits, indicating no immediate health risk from these two hormones for consumers. https://jfsh.tums.ac.ir/index.php/jfsh/article/view/629