HPLC法测定人血浆中头孢泊肟的浓度

Determination of content of cefpodoxime in human plasma by HPLC

目的建立测定人血浆中头孢泊肟酯的活性代谢产物头孢泊肟浓度的高效液相色谱法,并应用于头孢泊肟酯干混悬剂在健康人体内的药动学研究。方法采用沉淀蛋白法进行样品预处理,色谱柱为Agilent Zorbax SB-C18柱(150 mm×4.6 mm,5μm),流动相为乙腈-20 mmol.L-1磷酸二氢铵缓冲液(体积比为9∶91,含6.6 mmol.L-1三乙胺,磷酸调pH值至2.0),流速为1.0 mL.min-1,检测波长为254 nm,内标为头孢克洛。结果头孢泊肟浓度线性范围为0.102～6.528 mg.L-1,低、中、高3个质量浓度的提取回收率分别为92.0%、93.3%和92.1%,日内和日间精密度RSD(n=6)分别为3.3%、3.8%、2.0%和14.2%、2.9%、6.3%。头孢泊肟血浆样品室温放置2 h、预处理后室温放置24 h、经历1次及3次冷冻-解冻循环和-20℃冷冻情况下保存40 d均可保持稳定。结论此法适用于头孢泊肟酯药动学研究。

Objective To establish a high performance liquid chromatographic method for determination of the plasma concentration of cefpodoxime in healthy volunteers.Methods The samples were pretreated by protein precipitation.The chromatographic separation was achieved on a Agilent Zorbax SB-C18 column（150 mm×4.6 mm,5 μm）,with acetonitrile-20 mmol · L-1 ammonium dihydrogen phosphate（containing 6.6 mmol · L-1 triethylamine,pH 2.0）（V∶ V=9∶ 91）as the mobile phase at the flow rate of 1.0 mL · min-1.The detection wavelength was set at 254 nm,and the cefaclor was taken as the internal standard.Results A good linearity was obtained from 0.102 mg · L-1 to 6.528 mg · L-1 of cefpodoxime in human plasma.The recoveries were above 92.0%,the intra-day RSD and inter-day RSD were 3.3%,3.8%,2.0% and 14.2%,2.9%,6.3% in low,medium and high concentrations,respectively.Cefpodoxime plasma samples were found to be stable at room temperature for 2 h.Cefpodoxine was stable after three freeze/thaw cycles at room temperature for 24 h and long-term storage at-20 ℃ for fortydays.Conclusions The method is proven to be suitable for the pharmacokinetic study of cefpodoxime and monitoring its concentration for the clinical application.