目的:建立测定人血清中氨苄西林、丙磺舒浓度的高效液相色谱法.方法:(1)氨苄西林,以诺氟沙星为内标,血清样品用0.6%的高氯酸沉淀蛋白后进行测定;色谱柱为HigginsTM C18柱(250 mm ×4.6 mm,5μm);流动相为有机相(甲醇-乙腈-异丙醇,100∶2∶5,v/v):水相(0.01 mol/L磷酸二氢钾,pH 3.5)=23:77;检测波长210 nm;流速1.0 ml/min;柱温30℃.(2)丙磺舒,以缬沙坦为内标,血清样品用乙腈沉淀蛋白后进行测定;色谱柱为HigginsTM C18柱(250 mm×4.6 mm,5 μm);流动相为有机相(甲醇-乙腈,400∶275,v/v):水相(乙酸-水,1∶60,v/v)=77∶23;检测波长280 nm;流速1.0 ml/min;柱温30 ℃.结果:氨苄西林在0.14-11.2μg/ml浓度范围内线性关系良好(r=0.999 5),最低检测限为0.14 μg/ml;丙磺舒在0.20-16.0μg/ml浓度范围内线性关系良好(r=0.999 1),最低检测限为0.20μg/ml.结论:高效液相色谱法操作简便、灵敏、专一性好,适用于人体药动学研究.%Objective:To establish a method to determine the concentration of ampicillin and probenecid in human serum by high performance liquid chromatography ( HPLC) . Methods : ( 1 ) Ampicillin was detected by HPLC using norfloxacin as internal standard after sample was sedimented with 0. 6％ perchloric acid. HigginsTM C18 column( 250 mm x 4. 6 mm,5 μm) was used , the mobile phase consisted of organic phase ( methanol-acetonitrile-isopropanol, 100: 2: 5 , v/v) and aqueous phase ( 0. 01 mol/L potassium dihydrogen phosphate ,pH 3. 5) with a ratio of 23: 77 ,the flow rate was l. 0 ml/min, the detection wavelength was set at 210 nm , and column oven at 30 ℃ . (2) Probenecid was detected by HPLC using valsartan as internal standard after sample was sedimented with acetonitrile. HigginsTM C18 column ( 250 mm x 4. 6 mm,5 μm) was used, the mobile phase consisted of organic phase( methanol-acetonitrile, 400: 275 , v/v) and aqueous phase( acetic acid-water, 1: 60, v/v) with a ratio of 77: 23 ,the flow rate was l. 0 ml/min,the detection wavelength was set at 280 nm.and column oven at 30 ℃ . Results : Good linearity was obtained within the range of 0. 14 - 11. 2 μg/ml for ampicillin( r = 0. 999 5) , and 0. 20 - 16. 0 μg/ml for probenecid ( r = 0. 999 1). The mininum detectable concentrations for ampicillin , and probenecid were 0. 14 μg/ml and 0. 20 μg/ml, respectively. Conclusions : This method is simple , sensitive and specific , which can be applied to study the pharmacokinetics of ampicillin and probenecid.
展开▼
