Vomastová L, Miksík I, Deyl Z
Department of Analytical Chemistry, Institute of Chemical Technology, Prague, Czech Republic.
J Chromatogr B Biomed Appl. 1996 May 31;681(1):107-13. doi: 10.1016/0378-4347(95)00502-1.
A mixture of ten steroids was separated by microemulsion and micellar (SDS and glycodeoxycholate) electrokinetic chromatography systems. Separations were done on a 50 cm (to the detector) x 50 microns I.D. fused-silica capillary. Complete separation of all the test compounds in the micellar mode was obtained with glycodeoxycholate (50 mM) in 25 mM borate buffer, pH 6.5, as the micelle-forming agent. The best results, however, were obtained using microemulsion electrokinetic chromatography in which higher aliphatic alcohols were used as the microemulsion-forming modifiers. The system consisted of n-hexanol (0.81%), SDS (3.31%) and n-butanol (6.61%) in 20 mM phosphate buffer, pH 10.0 (89.28%, w/w). In the microemulsion mode, linear calibration for steroid standards was obtained in the concentration range 3 x 10(-4)-3 x 10(-5) mol 1(-1) with a detection limit of 1 pmol. The method was validated and applied to an 11 beta-hydroxysteroid dehydrogenase assay in tissues.
采用微乳液电动色谱系统和胶束电动色谱系统（十二烷基硫酸钠和甘氨胆酸盐）对十种甾体混合物进行了分离。分离在一根50 cm（至检测器）×50 μm内径的熔融石英毛细管上进行。以25 mM、pH 6.5的硼酸盐缓冲液中50 mM的甘氨胆酸盐作为胶束形成剂，在胶束模式下实现了所有测试化合物的完全分离。然而，使用微乳液电动色谱获得了最佳结果，其中使用高级脂肪醇作为微乳液形成改性剂。该系统由20 mM、pH 10.0的磷酸盐缓冲液（89.28%，w/w）中的正己醇（0.81%）、十二烷基硫酸钠（3.31%）和正丁醇（6.61%）组成。在微乳液模式下，甾体标准品在3×10⁻⁴ - 3×10⁻⁵ mol/L的浓度范围内获得线性校准，检测限为1 pmol。该方法经过验证并应用于组织中的11β - 羟基类固醇脱氢酶测定。
