本研究主要目的為利用串聯式液相層析質譜儀開發尿液檢體中類固醇運動禁藥（包含14種同化性類固醇與18種腎上腺糖皮質類固醇）之分析方法。本研究方法將尿液樣品經固相萃取、水解以及液相萃取後，注入液相層析系統，使用Discovery HS-C18逆相層析管柱（50 mm × 2.1 mm，3 μm），以移動相0.02%甲酸水溶液（移動相A）與甲醇（移動相B）進行梯度沖提；串聯式質譜分析器以電噴灑游離法之正電模式分析並採用多重反應監控模式收集分析結果。研究結果顯示，單一樣本分析時間為16分鐘，所有藥物相對滯留時間的同日間與異日間CV值皆小於1.0%，具有極佳的再現性，並以真實尿液檢品測試，確認本方法適用於一般運動禁藥之篩檢。本研究結論：所有藥物之偵測極限介於1～30 ng/mL，方法經驗確證具有一定穩定性及再現性，非常適合用於具有時效性，且在短期內有大量尿液檢體的運動禁藥篩檢分析上。

This study presents a general screening method for the detection of doping agents, including 16 anabolic steroids and 18 glucocorticoids in the human urine samples by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Sample preparation was done based on the single solid phase extraction cartridge of hydrolyzed human urine to produce extraction, and two kinds of solvent-methanol and dichloromethane mix solutions were collected individually. Then the assay was performed by LC-MS/MS in positive ionization mode using an electrospray ionization and multiple reaction monitoring as the acquisition mode. All compounds showed good reproducibility in terms of the relative retention times (both within-day and between-day precision CV% ＜ 1.0). The limit of detection (LOD) was in the range of 1-30 ng/mL for glucocorticoids and anabolic steroids. The overall performance of this method suggests that it is supposed to apply for a routine anti-doping analysis, especially when there is considerable number of samples during competitions.