Determination of 14 Anabolic Androgenic Steroids in Animal Derived Foods by QuEChERS-High Performance Liquid Chromatography-Tandem Mass Spectrometry

A method was developed for the determination of 14 anabolic androgenic steroids including trenbolone, boldenone and nandrolone in animal derived foods by liquid chromatography-tandem mass spectrometry. The samples were added to ammonium acetate buffer and enzymatic hydrolyzed by β-glucuronidase and arysulfatase. The hydrolyzates were extracted with 20 mL acetonitrile, and then 10 mL supernatant was degreased by 10 mL acetonitrile saturated hexane, purified with 50 mg C₁₈, 50 mg PSA and 300 mg neutral alumina powder including 300 mg MgSO₄. Then the analytes were separated by C₁₈ column with gradient elution using 0.1% formic acid and acetonitrile as the mobile phase, detected in positive multiple reaction monitoring (MRM) and quantified with matrix-matched external standards. The linear regression correlation coefficients for the compounds were all higher than 0.992 in the range from 0.1 to 10 ng/mL. The limits of detection and the limits of quantification were 0.3 and 1.0 μg/kg respectively. The recoveries at the addition levels of 1.0, 2.0 and 10 μg/kg were in the range of 75.7%~104.1%, with relative standard deviations (RSDs) of 3.60%~12.1%. This method had high effectivity, good reproducibility, simple operation and high efficiency, and could be used for the rapid determination needs of common anabolic androgenic steroids in foodborne stimulants in the Winter Olympic Games.