Iwata J, Suga T
Department of Clinical Biochemistry, Tokyo College of Pharmacy, Japan.
J Chromatogr. 1989 Jul 19;474(2):363-71. doi: 10.1016/s0021-9673(01)93929-9.
A new high-performance liquid chromatographic (HPLC) method with fluorescence detection is described for the direct determination of four serum 17-oxosteroid sulphates. Each serum sample was deproteinated with methanol, the methanol was evaporated and 17-oxosteroid sulphates in the residue were extracted with benzene as ion pairs in the presence of tetrapentylammonium ion. The ion pairs were labelled with dansylhydrazine and the hydrazones were separated by HPLC on a Capcell-Pak C8 (silicone polymer-coated silica gel modified with octyl groups) reversed-phase column using methanol-0.5% (w/v) sodium acetate-50% (v/v) acetic acid (57:42:1, v/v/v) as the mobile phase. The eluent was monitored with a fluorometric detector at an excitation wavelength of 330 nm and an emission wavelength of 540 nm.
本文描述了一种采用荧光检测的新型高效液相色谱（HPLC）方法，用于直接测定四种血清17-氧代类固醇硫酸盐。每个血清样品用甲醇进行脱蛋白处理，甲醇蒸发后，残留物中的17-氧代类固醇硫酸盐在四戊基铵离子存在下以离子对的形式用苯萃取。离子对用丹磺酰肼标记，腙在Capcell-Pak C8（用辛基改性的硅胶聚合物涂层硅胶）反相柱上通过HPLC分离，流动相为甲醇-0.5%（w/v）醋酸钠-50%（v/v）醋酸（57:42:1，v/v/v）。用荧光检测器在激发波长330nm和发射波长540nm下监测洗脱液。
