Automated and sensitive determination of four anabolic androgenic steroids in urine by online turbulent flow solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry: a novel approach for clinical monitoring and doping control.
A novel method for automated and sensitive analysis of testosterone, androstenedione, methyltestosterone and methenolone in urine samples by online turbulent flow solid-phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry was developed. The optimization and validation of the method were discussed in detail. The Turboflow C18-P SPE column showed the best extraction efficiency for all the analytes. Nanogram per liter (ng/L) level of AAS could be determined directly and the limits of quantification (LOQs) were 0.01 ng/mL, which were much lower than normally concerned concentrations for these typical anabolic androgenic steroids (AAS) (0.1 ng/mL). The linearity range was from the LOQ to 100 ng/mL for each compound, with the coefficients of determination (r(2)) ranging from 0.9990 to 0.9999. The intraday and interday relative standard deviations (RSDs) ranged from 1.1% to 14.5% (n=5). The proposed method was successfully applied to the analysis of urine samples collected from 24 male athletes and 15 patients of prostate cancer. The proposed method provides an alternative practical way to rapidly determine AAS in urine samples, especially for clinical monitoring and doping control.
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
Feng
F
Guo
National Institute of Sports Medicine, Beijing 100029, China.
Jing
J
Shao
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
Qian
Q
Liu
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China. Electronic address: jbshi@rcees.ac.cn.
Jian-Bo
JB
Shi
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
Gui-Bin
GB
Jiang
D045930:Anabolic Agents / Q000652:urine*; D001331:Automation; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D004300:Doping in Sports; D004867:Equipment Design; D006801:Humans; D057230:Limit of Detection; D008297:Male; D013058:Mass Spectrometry; D011471:Prostatic Neoplasms / Q000652:urine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D013256:Steroids / Q000652:urine*; D015813:Substance Abuse Detection / Q000379:methods*; D053719:Tandem Mass Spectrometry; D016482:Urinalysis / Q000379:methods
D045930:Anabolic Agents; D013256:Steroids
Anabolic androgenic steroids; Clinical monitoring; Doping control; Online turboflow solid-phase extraction; Urine