高效毛细管电泳法测定人血浆育亨宾浓度

Analysis of Yohimbine in Human Plasma by Non-aqueous Capillary Electrophoresis (NCAE)

目的建立非水毛细管电泳法(NCAE)测定人血浆育亨宾浓度的新方法。方法运用非水毛细管电泳方法,熔融石英毛细管(75μm×50cm),缓冲液为30mmol·L-1醋酸铵的甲醇-乙腈液(4:1,V/V,含1%冰醋酸),检测波长:220nm,分离电压:25kV,柱温25℃,用0.45μm微孔滤膜过滤后进样,压力进样:50kPa×5s,采用峰面积内标法定量。结果方法最低检测浓度为0.01μg·mL-1,线性范围0.0548～5.4800μg·mL-1,r=0.9991,线性关系良好。日内RSD为2.81%～5.22%,日间RSD为4.72%～6.52%。结论该方法测定人血清中育亨宾含量快速、准确和灵敏。

Objective To develop a rapid method for determination of yohimbine in human plasma by non-aqueous capillary electrophoresis （NCAE）. Methods The separation conditions of NCAE were 30 mmol ·mL^-1 ammonium acetate in methanol-acetonitrile （4 ： 1, V/V , added 1% glacial acetic acid）as running buffer, UV detection was set at 220 nm, separation voltage was 25 kV, column temperature 25 ℃and pressure injection was 50 kPa × 5 s. Internal calibration curve of peak area was selected to quantify. Results The limits of detection was 0.01 μg·mL^-1 The concentration of yohimhine showed a linear plot in the range of 0. 0548-5.48 μg·mL^-1 （ r = 0.999 1 ） . Inter-day and intra-day precisions expressed as the relative standard deviation （RSD） for the method was 2.81% -5.22% and 4.72% -6.52% , respectively. Conclusion This method is rapid and accurate for the quantitative analysis of yohimbine in human plasma.