ダンシルヒドラジンを用いる生体試料中ケトステロイドのけい光高速液体クロマトグラフィー : 臨床分析への応用

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The determination of ketosteroids in biological fluids by a fluorescence highperformance liquid chromatographic procedure and the application of this method to clinical analysis of steroidal compounds in the human blood and urine are described. Corticoids are extracted with methylene chloride from 0.5 ml of plasma or 1 ml of urine, and 17-ketosteroids are extracted with methylene chloride after enzymatic hydrolysis or solvolysis (1 ml of urine or 0.1 ml of serum). Δ^4-3-Ketosteroids and 17-ketosteroids are labelled with dansyl hydrazine in HCl-EtOH solution and in trichloroacetic acid-benzene solution, respectively, and chromatographed on a microparticulate silica gel column by use of CH_2Cl_2-EtOH-H_2O mobile phase systems with a fluorophotometric detector at 350 nm (excitation) and 505 nm (emission). Comparisons with radioimmunoassay gave correlation coefficients of 0.978 (n=36) and 0.964 (n=81) for plasma cortisol and serum dehydroepiandrosterone sulfate, respectively. Metabolite of carbamazepine did not interfere with the analysis of urinary free cortisol. The serum concentrations of cortisol, 11-deoxycortisol and dehydroepiandrosterone sulfate after a single oral administration of 1.5 g metyrapone to human subjects were determined and the serum 11-deoxycortisol could be measured by this method as a rapid metyrapone test. The method is thus useful for the clinical analysis of ketosteroids in biological fluids.
公益社団法人日本薬学会の論文
1980-09-25

ダンシルヒドラジンを用いる生体試料中ケトステロイドのけい光高速液体クロマトグラフィー : 臨床分析への応用

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