胶束电动色谱法快速测定鱼血液中的类固醇激素

Fast determination of steroid hormones in fish blood with micellar electrokinetic capillary chromatography

建立了一种同时分离测定鱼血液中皮质醇、雌二醇及睾酮含量的胶束电动色谱法.研究了缓冲溶液浓度、添加剂含量、pH、分离电压及温度等条件对分离的影响,得到了最佳分离条件,即:缓冲溶液组成为12mmol/L硼砂+60 mmol/L十二烷基硫酸钠+6 mmol/Lβ-环糊精+8%乙腈,pH 9.5;检测波长为225 nm,分离电压为25 kV,分离温度为25℃;采用毛细管短端进样(毛细管有效长度仅为10 cm)、反向分离模式.3种类固醇激素在4 min内达到基线分离,且分析物在1.0～100.0μg/mL呈良好的线性关系,相关系数大于0.993 2.该方法用于鱼血液中类固醇激素的检测,回收率为84.60%～113.30%,满足生物样品检测的要求.

A novel micellar electrokinetic capillary chromatography （MEKC } method was developed for sepa- ration and detection of cortisol, 17β-estradiol and testosterone in fish blood. The factors affecting separation efficiency such as concentration of buffer solution and surfactant, additives, pH, separation voltage and tem- perature were investigated. The optimal conditions were concluded as follows： 12 mmol/L borax ＋ 60 mmoL/L sodium dodecyl sulfate （SDS） ＋ 6 mmolfLβ-cyelodextrin （β-CD） ＋ 8% （v/v） acetonitrile, pH 9.5 , detection wavelength of 225 nm, separation voltage and temperature of 25 kV and 25 ℃, respectively. Under the optimal conditions, the analytes could be base-line separated within 4 min using reverse separation mode with capillary length of 10 era. Good linear relationship were observed in the range of 1.0 - 100.0μg/mL with correlation coefficient larger than 0. 993 2. The proposed method had been successfully applied tothe determination of steroid hormones in fish blood with recoveries in range of 8g. 60% ～113.30%.