Kawasaki T, Maeda M, Tsuji A
J Chromatogr. 1982 Oct 8;232(1):1-11. doi: 10.1016/s0378-4347(00)86001-6.
A method is described for the determination of urinary 17-hydroxycorticosteroids using fluorescence high-performance liquid chromatography. After enzymatic hydrolysis, 17-hydroxycorticosteroids were extracted using an Extrelut column and then labeled with Dns-hydrazine in hydrochloric acid-ethanol solution. The labeled steroids were chromatographed on a microparticulate silica gel column, the mobile phase was dichloromethane-ethanol-water (900:60:40). The eluate was monitored on a fluorophotometer at 365 nm (excitation) and 505 nm (emission). Linearity of the fluorescence intensities (peak heights) of various 17-hydroxycorticosteroids were obtained between 60 pg and 20 ng. The assay was sensitive, precise and accurate. Comparison with the results obtained by radioimmunoassay gave correlation coefficients of 0.932 for tetrahydrocortisol and 0.930 for tetrahydrocortisone. The proposed method is clinically useful for the routine analysis of urinary 17-hydroxycorticosteroids.
描述了一种使用荧光高效液相色谱法测定尿中17-羟皮质类固醇的方法。酶解后，使用埃特拉路特柱提取17-羟皮质类固醇，然后在盐酸-乙醇溶液中用丹磺酰肼标记。将标记的类固醇在微粒硅胶柱上进行色谱分析，流动相为二氯甲烷-乙醇-水（900:60:40）。在荧光光度计上于365nm（激发）和505nm（发射）处监测洗脱液。各种17-羟皮质类固醇的荧光强度（峰高）在60pg至20ng之间呈线性关系。该测定法灵敏、精确且准确。与放射免疫分析法所得结果比较，四氢皮质醇的相关系数为0.932，四氢可的松的相关系数为0.930。所提出的方法在临床上可用于尿中17-羟皮质类固醇的常规分析。
