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  • Title: Determination of urinary free cortisol and cortisone by sequential thin layer and high-performance liquid chromatography.
    Author: Lewbart ML, Elverson RA.
    Journal: J Steroid Biochem; 1982 Aug; 17(2):185-90. PubMed ID: 6287108.
    Abstract:
    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.
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