Analysis of corticosteroids in urine by HPLC and thermospray LC/MS.
The method for simultaneous determination and confirmation of nine corticosteroids in urine by high-performance liquid chromatography with a diode array detector and thermospray liquid chromatography-mass spectrometry was studied. HPLC was performed on a C18 column with a gradient mobile phase system of water and acetonitrile. The calibration curve was linear from 20 ng/mL to 1.0 micrograms/mL for each corticosteroid, and the detection limit was 10 ng/mL in 5 mL of urine. The extraction recovery of each corticosteroid from the spiked urine was equal to or greater than 85% by liquid-liquid extraction (LLE) at pH 9 using diethyl ether and approximately 79% or more by solid-phase extraction (SPE) with a Sep-Pak C18 cartridge. The mass spectra obtained with the positive ion mode showed protonated molecular species [M + H]+, ammonium adduct ion [M + NH4]+, [MH-60]+, [MH-30]+, and [MH-18]+. The LC/MS detection limits ranged from 10 to 50 ng in the scan mode and from 1 to 5 ng in the selected ion monitoring (SIM) mode.
Doping Control Center, Korea Institute of Science and Technology, Cheongryang, Seoul.
S J
SJ
Park
Y J
YJ
Kim
H S
HS
Pyo
J
J
Park
D000305:Adrenal Cortex Hormones / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D004300:Doping in Sports; D005938:Glucocorticoids / Q000652:urine*; D006801:Humans; D006908:Hydroxyprogesterones / Q000652:urine*; D013058:Mass Spectrometry; D015813:Substance Abuse Detection / Q000379:methods*
D000305:Adrenal Cortex Hormones; D005938:Glucocorticoids; D006908:Hydroxyprogesterones; C030228:11-hydroxyprogesterone