HPLC法测定阿莫西林舒巴坦酯片人体血药浓度

    【关键词】 阿莫西林舒巴坦酯片；高效液相色谱法；血药浓度

      To detect human blood concentration of amoxicillin and sulbactam with HPLC

    【Abstract】 Objective To set up the detection method of human blood concentration of amoxicillin and sulbactam. Methods The chromatographic column was Spberiserb ODS(φ3.9mm×200mm.5μm ),and the mobile phase was acetonitrile- 0.068mol/L KH2PO4(4.5:95.5),the flow rate was 0.8ml/min,the detection wavelength was 225nm,and the column temperature was 25±0.1 ℃.We detected the human blood concentration of amoxicillin,the chromatographic colum was Hypersil C18(φ4.6mm×200mm 5μm),the mobile phase was acetonitrile-0.068mol/L KH2po4(4.5:95.5),the flow rate was 1.2ml/min,the detection wavelength was 210nm, and the column temperature was 25±0.1 ℃.We detected the human blood concentration of sulbactam. Results We found that amoxicillin and sulbactam were under the condition of selected chromatographic separation, the resolution of amoxicillin and sulblactam and the impurity in the blood plasma was favorable.The resolution was <1.5,the human blood concentration of amoxicillin was 0.125～10μg/ml, and the linear regrossion was favorable,r=0.9998.The human blood concentration of sulbactam was 0.10～10μg/ml,and the linear regrossion was favorable,r=0.9999.The average recovery of amoxicillin and sulbactam were 96.25%and 102.3%.The two kinds of components were RSD<10%in a day or next.Conclusion The study shows that we can detect the human blood concentration of amoxicillin and sulbactam with HPLC,and the method has very good accuracy,precision and sensitivity.It is accord with demands of drug detection.

    【Key words】 amoxicillin and sulbactam;liquid chromatography;blood concentration

    阿莫西林舒巴坦酯是由阿莫西林和舒巴坦以共价键结合形成的化合物，阿莫西林为广谱的β-内酰胺类青霉素，舒巴坦是β-内酰胺酶抑制剂，无抑菌活性，能够抑制细菌产生的Ⅱ型～Ⅳ型β-内酰胺酶，使阿莫西林免受酶的水解灭活，从而增加阿莫 西林对细菌的敏感性。该药口服后在胃肠上壁被非 特异酯酶分解为阿莫西林和舒巴坦，并吸收进入人体血液循环［1］。临床上主要用于治疗上、下呼吸道感染和皮肤、软组织感染及性病等［2］。本文研究阿莫西林舒巴坦片口服后血浆中阿莫西林和舒巴坦药物浓度测定方法，为该制剂的人体药代动力学和生物学等效性研究及临床用药提供参考。

    1 材料与方法

    1.1 药品与试剂 阿莫西林对照品（中国药品生物制品检定所，含量86.2%、批号0409-9706）；舒巴坦对照品（中国药品生物制品检定所，含量99.8%、批号0430-9501）；阿莫西林舒巴坦酯片（珠海联邦制药有限公司中山分厂生产，每片含阿莫西林250mg、舒巴坦250mg，批号030418）；乙腈为色谱纯；乙醚、磷酸二氢钾、盐酸高氯酸为分析纯；水为超纯水。

    1.2 仪器 waters高效液相色谱分析系统（包括717自动进样器、510低压梯度泵、996PDA二极管检测器，2010色谱管理系统），waters高效液相色谱分析系统（包括717自动进样器、600低压梯度泵、996PDA二极管检测器，2010色谱管理系统）；液体快速混合仪（北京长安仪器厂）；普通离心机（北京医用离心机厂）。

    1.3 色谱条件

    1.3.1 阿莫西林色谱条件 Spberserb ODS(Φ3.9mm×200mm,5μm),C18预柱；流动相：乙腈-0.068mol/L KH2PO4(4.5:95.5);流速：0.8ml/min;检测波长：225nm；柱温：（25±0.1）℃;灵敏度1.0AUFS。

    1.3.2 舒巴坦色谱条件 Hyperisil C18 (Φ4.6mm×200mm,5μm)，C18预柱；流动相：乙腈-0.068mol/L KH2PO4(4.5:95.5);流速：1.2ml/min;检测波长：210nm；柱温：（25±0.1）℃;灵敏度0.5AUFS。

    1.4 受试对象与实验设计 健康男性自愿受试者20人，汉族，年龄23～25岁，体重60～70kg,身高168～175cm。经心电、血压、血尿常规及肝肾功能检查均正常并无青霉素过敏史，签署知情同意书者入选，实验前2周禁服任何药物。实验前空腹12h，于早8：00时用250ml温开水送服药物。服药后4h统一食低蛋白低脂肪午餐，观察病房准备危急情况的抢救措施。受试者口服阿莫西林舒巴坦酯片2片（含阿莫西林和舒巴坦均为500mg），于服药前（0h）与服药后10、20、30、45、60、90、120、180、240、420min采静脉血3ml，肝素抗凝，离心取血浆，置于-20℃冰箱中备用。转贴于 酷文网-论文下载中心 http://www.coolwen.net