Kawasaki T, Maeda M, Tsuji A
J Chromatogr. 1979 Jun 11;163(2):143-50. doi: 10.1016/s0378-4347(00)81457-7.
A method is described for the determination of cortisol in human plasma and urine by high-performance liquid chromatography using fluorophotometric detection. After extraction with methylene chloride, cortisol is labelled with dansyl hydrazine, and then separated by high-performance chromatography. The eluate is monitored by a fluorophotometer at 350 nm (excitation) and 505 nm (emission). The optimum conditions for the determination, such as HCl and dansyl hydrazine concentrations, reaction time and reaction temperature, and for the eluent of high-performance liquid chromatography, are discussed. Linearity of the fluorescence intensity (peak height) with the amount of cortisol was obtained between 0.5 and 60 ng. The recoveries for 50 and 100 ng of added cortisol were 98.7 and 95.4% for plasma, and 96.4 and 90.6% for urine, respectively. Comparison with a radioimmunoassay gave a correlation coefficient of 0.978. The proposed method is suitable for the routine analysis of cortisol in plasma and urine.
描述了一种用高效液相色谱法结合荧光光度检测来测定人血浆和尿液中皮质醇的方法。用二氯甲烷萃取后，皮质醇用丹磺酰肼标记，然后通过高效色谱法分离。洗脱液用荧光光度计在350nm（激发）和505nm（发射）处监测。讨论了测定的最佳条件，如盐酸和丹磺酰肼的浓度、反应时间和反应温度，以及高效液相色谱的洗脱液。荧光强度（峰高）与皮质醇量在0.5至60ng之间呈线性关系。添加50和100ng皮质醇时，血浆中的回收率分别为98.7%和95.4%，尿液中的回收率分别为96.4%和90.6%。与放射免疫分析法比较，相关系数为0.978。所提出的方法适用于血浆和尿液中皮质醇的常规分析。
