김현석 (식품의약품안전평가원)

김현경 (식품의약품안전평가원)

류금주 (식품의약품안전평가원)

김현석 (식품의약품안전평가원)

서소운 (식품의약품안전평가원)

김애슬 (식품의약품안전평가원)

김형수 (식품의약품안전평가원)

김현경 (식품의약품안전평가원)

Anabolic androgenic steroids (AAS) are testosterone-derived compounds that promote muscle growth by stimulating protein synthesis in muscle tissue. However, abuse of AAS has been known to be associated with serious health problems such as liver dysfunction and cardiovascular damage. Despite being legally prohibited, illicit production and distribution of AAS continue on the black markets. In this study, we developed a rapid, sensitive, and comprehensive analytical method employing ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and quadrupole time-of-flight mass spectrometry (LC-QTOF/MS) for the simultaneous detection of 45 AAS. LC-QTOF/MS was used to characterize the fragmentation patterns of each compound and to inform the selection of product ions for multiple reaction monitoring in LC-MS/MS method development. The LC-MS/MS method was validated for specificity, with a limit of detection (LOD) ranging from 0.13 to 16.70 ng/mL and a limit of quantification (LOQ) between 0.40 and 50.00 ng/mL across both solid and liquid matrices. The method showed excellent linearity (R² ≥ 0.999), recovery (82.5–112.2% for solids, 80.1–116.5% for liquids), intra-day accuracy (90.5–118.8% and 84.7–118.0%), inter-day accuracy (80.5–119.2% and 81.2–117.5%), intra-day precision (0.1–6.0% RSD and 0.1–6.5% RSD), and inter-day precision (0.1–5.2% RSD and 0.4–8.0% RSD), in accordance with ICH guidelines.