中空纤维液相微萃取结合高效液相色谱法测定大鼠血浆中小檗碱含量研究

Hollow fiber based liquidp hase microextraction combined with high-performance liquid chromatography for study of the pharmacokineticsof berberine in rat plasma

目的建立基于中空纤维液相微萃取结合HPLC测定大鼠血浆中小檗碱含量的方法。方法采用中空纤维液相微萃取技术对大鼠血浆样品进行预处理，优化影响中空纤维液相微萃取的主要因素，在最佳萃取条件下（即给出相和接收相的pH值分别为12．0、2．0，萃取时间为20min，搅拌速度为800r／min，盐浓度为10％），大鼠血浆中小檗碱的富集因子为347。同时建立HPLC法测定大鼠血浆中小檗碱含量的分析方法。结果大鼠血浆中小檗碱在浓度10～1000ng／ml范围内线性良好（r2≥0．9992），最低定量浓度（S／N=10）为10ng／ml，最低检测限（S／N=3）为3．3ng／ml，日内精密度、日间精密度和稳定性试验的RSD值均〈6．33％，平均加样回收率为（96．7±3．82）％，RSD为4．82％。结论采用中空纤维液相微萃取法代替传统萃取技术萃取大鼠血浆中小檗碱，具有高效、环保，方法精密度、重复性好等优点，可用于小檗碱体内的含量测定。

Objective A new hollow fiber liquid phase microextraction combined with high-performance liquid chromatography was developed to determine the concentration of berberine in rat plasma. Methods Parameters that affect the hollow fiber liquid phase microextraction processes were investigated and optimized. The optimized conditions were pH of donor and acceptor phases 12 and 2.0, respectively; extraction time 20 min; stirring speed 800 rpm; and addition of 10 % （w/v） salt. Under the optimized conditions, the preconcentration factor for berberine was 347. Moreover, a rapid and sensitive method was developed to determine berberine in rat plasma by HPLC. Results The calibration curve for berberine was linear in the range of 10-1000 ng/ml （r2≥0.9992）. The limit of quantitation for the analyte was 10 ng/ml （S/N=10）. The limit of detection for the analyte was 3.3 ng/ml （S/N=3）. The intra-day and inter-day precision, and stability （RSD） were less than 6.3%. The average recovery was 96.7% ± 3.82%, and RSD was 4.82%. Conclusions The method is efficient, green, accurate and repeatable. It can be applied in determination of berberine in rat plasma.