Determination of urinary free cortisol and cortisone by sequential thin layer and high-performance liquid chromatography.
A specific, rapid and sensitive method for urinary free cortisol and cortisone utilizing sequential thin layer (TLC) and high-performance liquid chromatography (HPLC) was developed. After addition of prednisone as the internal standard to 1 ml of urine, extraction of the steroids was accomplished by automated sorption on and desorption from a styrenedivinylbenzene copolymeric resin cartridge. Further purification was carried out by TLC. Quantitation of the recovered steroids at 254 nm was achieved during HPLC in a reverse phase system with 21-deoxycortisone as the external standard. When the values of urinary free cortisol both from normal subjects and patients were compared with those obtained by a current RIA procedure, the greater specificity of the new method was clearly demonstrated. Simultaneous measurement of both urinary free cortisol and cortisone in various clinical states appears to offer a more complete index of adrenocortical function than urinary free cortisol alone.
D000324:Adrenocorticotropic Hormone; D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002855:Chromatography, Thin Layer / Q000379:methods; D003348:Cortisone / Q000652:urine*; D005260:Female; D006801:Humans; D006854:Hydrocortisone / Q000652:urine*; D008297:Male; D008832:Microchemistry; D011863:Radioimmunoassay / Q000379:methods; D012016:Reference Values
D000324:Adrenocorticotropic Hormone; D003348:Cortisone; D006854:Hydrocortisone