效应面法优化浊点萃取-HPLC测定小鼠脑组织中蛇床子素含量

Determination of Osthole in Mouse Brain Tissue Using HPLC Using Cloud Point Extraction with Optimization by Response Surface Methodology

目的建立基于浊点萃取同时测定小鼠脑组织中蛇床子素含量的HPLC法。方法以非离子表面活性剂Triton X-114为浊点萃取剂,考察了Triton X-114浓度、NaCl浓度、萃取温度和时间等对蛇床子素萃取有影响的因素,并进一步采用效应面法分析优化实验条件。同时建立了HPLC法测定小鼠脑组织中蛇床子素含量的分析方法。结果蛇床子素在最佳萃取条件下的萃取率为93.8%。RSD≤5.6%,加样回收率为97.2%。结论采用浊点萃取法代替传统有机溶剂萃取小鼠脑组织中蛇床子素,操作简便,萃取率高,方法重复性好,可用于蛇床子素体内药代动力学的研究。

Objective To establish a new method based on cloud-point extraction（CPE） to determine osthole in mouse brain tissue by reversed phase HPLC.Methods The non-ionic surfactant Triton X-114 was chosen as the extract solvent.Response surface methodology was employed to further analyze and optimize the parameters affecting the CPE efficiency,e.g.,concentration of TritonX-114 and NaCl,temperature and time.Moreover,a rapid and sensitive method was developed to determine ostholein mouse brain tissue by HPLC.Results Under the optimum conditions,the extraction rate of osthole in CPE was up to 93.8%.The method was shown to be reproducible and reliable with RSD≤5.6% and mean recovery was up to 97.2%.Conclusion The CPE technique is a low-cost,simple and sensitive method with high extraction effect.This method has successfully been applied to the pharmacokinetic study of osthole.