响应面优化浊点萃取联合HILIC测定大鼠血清中四种抗艾滋病药

Response Surface Optimization of Cloud Point Extraction-HILIC Determination of Four Anti-AIDS Drugs in Rat Serum

[目的]基于浊点萃取技术(CPE)联合亲水作用色谱法(HILIC)同时测定大鼠血清中4种抗艾滋病药。[方法]以TritonX-114作为萃取介质,用响应面法进一步优化CPE相关参数,对萃取的样品,用HILIC法分别测定其含量。[结果]4种抗艾滋病药在5%(w/v)TritonX-114,0.3mol/LNaCl,pH为5.0,40℃水浴平衡20min时,萃取率均达85.0%以上,与模型预测值接近;对于富集的样品,以Ze month-CN(250 mm×4.6 mm,5μm)为固定相,流动相为(甲醇∶乙腈∶醋酸铵缓冲液=5590),柱温35℃,检测波长275nm,流速0.5mL/min分离时,4种分析物在0.01~10.0μg/mL,线性良好,加样回收率大于75.0%,RSD≤5.0%。[结论]建立的该方法,操作简单绿色,能对大鼠血清中4种抗艾滋病药较好的富集,可同时快速、准确测定其含量,为其临床血药浓度监测提供新的方法。

【Objective】Cloud point extraction(CPE)combined with hydrophilic interaction(HILIC)was em⁃ployed to simultaneously determine the anti-AIDS drugs in the serum of rats.【Methods】Triton X-114 was used as ex⁃traction medium to extract four nucleoside antiviral drugs from rat serum.Response surface methodology was employed to further optimize CPE parameters.The content of four anti-AIDS drugs in rat serum was simultaneously determined by HILIC method.【Results】The optimized conditions were as follows:5%(w/v)Triton X-114,0.3 mol/L NaCl,PH 5.0,the water-bath equilibrium 20 min at 40℃.Under optimized extraction conditions,the extraction rates of the four anti-AIDS drugs were all over 85.0%,which was pretty close to the predicted result.The extracted samples were analyzed under the optimal chromatographic conditions.The recovery rate was over 75.0%and RSD was less than 5.0%.The optimized chromatographic conditions were as follows:Ze month-CN(250×4.6 mm,5 micron)as stationary phase,mobile phase(methanol:acetonitrile:ammonium acetate buffer)(5∶5∶90),column temperature 35℃,detection wavelength 275 nm,flow rate 0.5 mL/min.【Conclusion】The developed method of CPE combined with HILIC can enrich four anti-AIDS drugs in rat serum,which is simple,environment-friendly and can be used to accurately determine the blood concentration of four anti-AIDS drugs in rat serum,providing a new method for clinical blood concentration monitoring.