Development and Method Validation for Testosterone and Epitestosterone in Human Urine Samples by Liquid Chromatography Applications | Zendy

R. GonzaloLumbreras | Zendy; D. Pimentel-Trapero | Zendy; R. IzquierdoHornillos | Zendy

Open Access

Development and Method Validation for Testosterone and Epitestosterone in Human Urine Samples by Liquid Chromatography Applications

Author(s) -

R. GonzaloLumbreras,

D. Pimentel-Trapero,

R. IzquierdoHornillos

Publication year - 2003

Publication title -

journal of chromatographic science

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.362

H-Index - 56

eISSN - 1945-239X

pISSN - 0021-9665

DOI - 10.1093/chromsci/41.5.261

Subject(s) - epitestosterone , chromatography , chemistry , extraction (chemistry) , urine , solid phase extraction , liquid–liquid extraction , high performance liquid chromatography , detection limit , population , androgen , biochemistry , demography , sociology , hormone

An isocratic high-performance liquid chromatographic method for the determination of testosterone (T) and epitestosterone (ET) in human urine using liquid-liquid or solid-phase extraction (SPE) is developed and validated. The optimum separation is achieved using a Hypersil C(18) column, water-acetonitrile (57:43, v/v) as the mobile phase and UV-absorbance detection at 245 nm. The recoveries obtained for T and ET in liquid-liquid and SPE demonstrate that these procedures are interchangeable. Quantitation limits for T and ET are 8.6 and 5.4 ng/mL using solvent extraction and 7.3 and 5.7 ng/mL using SPE, respectively. The proposed method is used to evaluate the urinary T, ET, and the T/ET ratio for a healthy male population using liquid-liquid extraction, and the T and ET excretion profile for nine healthy men using SPE.

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