人血浆中阿莫西林-克拉维酸浓度的测定

Determination of concentration of amoxycillin-clavulanic acid in human plasma by HPLC method

目的 :建立阿莫西林 -克拉维酸血药浓度的HPLC法。方法 :固定相为C18对称反相色谱柱 ;流动相为磷酸 -四丁基氢氧化铵缓冲液∶甲醇 (4 8∶5 2 ,V/V) ;流速 0 8mL·min-1;荧光检测波长 :Ex 386nm ,Em 4 6 0nm ;血浆标本以高氯酸去蛋白 ,再以苯甲醛荧光衍生后上柱分析 ,进样量 5 0 μL。 结果 :阿莫西林和克拉维酸的色谱峰分离良好 ,最低检测质量浓度分别为 0 0 0 9和 0 0 1mg·L-1;线性范围分别为 0 12 5～ 2 5和 0 0 6 2 5～ 8mg·L-1;平均日内、日间RSD均 <9% ,提取回收率近 10 0 %。结论 :本方法适用于阿莫西林

AIM: To establish a HPLC method for the determination of amoxycillin-clavulanic acid in human plasma. METHODS: Solid phase was symmetry C 18, mobile phase was phosphatic-tetrabutylammonium hydroxide buffer∶methanol(48∶52,V/V). Flow rate was 0.8 mL·min -1; detecting wavelength: Ex 386 nm, Em 460 nm. After plasma protein was precipitated by perchloric acid, the suspension was derivatized with benzal dehyde at 99 ℃ before the derivatized sample was eluted and 50 μL injected. RESULTS: The minimum detective concentration of amoxycillin and clavulanic acid was 0.009 and 0.01 mg·L -1 respectively,and concentrations range of standard curve was from 0.125 to 25 and 0.062 5 to 8 mg·L -1,respectively. The mean RSD within-day and between day were less than 9%. Recovery rates of plasma extraction were near 100%. CONCLUSION: The method adapts to the pharmacokinetic study and the bioequivalence of amoxycillin-clavulanic acid.