College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, P.R. China.
Biomed Chromatogr. 2020 Jun;34(6):e4824. doi: 10.1002/bmc.4824. Epub 2020 Mar 18.
Electrokinetic supercharging, a convenient and powerful online preconcentration technique in capillary electrophoresis, was introduced and evaluated for the determination of two alkaloids, berberine and jatrorrhizine, in mice fecal samples for the first time. The method depended on using a bare fused silica capillary (50 cm × 50 μm i.d.) and applying the voltage of 25 kV with UV detection at 205 nm. Parameters that affect the separation and preconcentration efficiency have been optimized. The optimum conditions used were as follows: background electrolyte consisting of 40mM sodium dihydrogenphosphate containing 30% methanol (v/v); hydrodynamic injection of 20mM KCl (50 mbar × 150 s) as the leading electrolyte; electrokinetic injection of the sample (+15 kV, 120 s) followed by the hydrodynamic injection of 30mM dodecyl trimethyl ammonium chloride (50 mbar × 12 s) as the terminating electrolyte. The results showed that the detection sensitivity of berberine and jatrorrhizine was, respectively, improved up 2740- and 2928-fold compared with normal injection, providing limits of detection lower than 3 ng/mL with good repeatability in areas (relative standard deviation < 3%). In summary, the developed method proved its ability in analyzing trace alkaloids in complicated biological samples.
电动强化,毛细管电泳中一种简便、强大的在线浓缩技术,首次被引入并评估用于测定小鼠粪便样品中的两种生物碱,小檗碱和黄连碱。该方法依赖于使用裸熔融二氧化硅毛细管(50cm×50μm id),并在 205nm 处进行紫外检测施加 25kV 的电压。已经优化了影响分离和浓缩效率的参数。最佳条件如下:背景电解质由 40mM 磷酸二氢钠组成,含有 30%甲醇(v/v);作为先导电解质的 20mM KCl 的液压注射(50mbar×150s);作为终止电解质的样品的电动注射(+15kV,120s),随后进行 30mM 十二烷基三甲基氯化铵的液压注射(50mbar×12s)。结果表明,与正常进样相比,小檗碱和黄连碱的检测灵敏度分别提高了 2740 倍和 2928 倍,检测限低于 3ng/mL,面积重复性好(相对标准偏差<3%)。总之,所开发的方法证明了其在分析复杂生物样品中痕量生物碱的能力。