反相高效液相色谱法测定人血浆中薁磺酸钠浓度

Determination of azulene concentration in human plasma by RP-HPLC

目的 建立艹奥 磺酸钠血药浓度测定的高效液相色谱法。方法 以对二甲氨基苯甲醛为内标 ,甲醇为血浆蛋白沉淀液 ,上清液经 40℃真空干燥恒温箱挥干 ,10 0 μl流动相溶解 ,2 0 μl进样。以WatersymmetryC18柱 ( 3 9× 15 0mm ,5μm)为分析柱。流动相为甲醇∶0 0 5mol/L磷酸缓冲液 =5 7∶43 (pH 7 0 ) ,流速 1 0ml/min ,柱温 2 5℃ ,检测波长 2 92 5nm。结果 血药浓度线性范围 2 5～ 64 0 0ng/ml (n =9,r =0 9966) ,最低检测浓度 2 5ng/ml(S/N >3 ) ,相对回收率 90 5 8%～111 0 0 % ,日内和日间RSD分别为 0 2 %～ 5 4%和 2 4%～ 6 0 %。结论 所建方法简便、准确、重现性好、灵敏度高 ,适用于艹奥 磺酸钠血药浓度的实际测定。

Objective To establish HPLC method for the determination of azulene (AZU) concentration in human plasma. Methods ρ-(dimethylamino)-benzaldehyde was employed as an internal standard (IS) and methanol as a protein precipitation. AZU and IS were separated on the water symmetry C18 column with methanol at the dose of 0.05 mol/L phosphate buffer of pH 7.0 (57∶43) as the mobile phase. The flow rate was 1 ml/min and the detection wavelength was set at 292.5 nm. The column temperature was 25 ℃. Results The linear rang of calibration curve was within drug plasma concentrations of 25-6 400 ng/ml (r=0.996 6). The lower limit of quantification (S/N≥3) was 25 ng/ml. The intra- and inter-assay coefficients of variation ranged from 0.2% to 5.4% and 2.4% to 6.0% . The recovery ranged from 90.58% to 111.00%. Conclusion The established method with the advantages of simplicity, precision, high sensitivity, and high repeatability, is suitable for the detection of azulene concentration in human plasma.