HPLC法测定人血浆中万古霉素浓度及其药动学研究

Determination of vancomycin concentration in human plasma by HPLC and study on its pharmacokinetics

目的建立HPLC法测定人血浆中万古霉素浓度的方法,并研究其在健康人体内的药动学。方法以万古霉素作为外标进行定量,血浆样品经10%高氯酸沉淀后,采用Agilent ZORBAX SB-C18色谱柱(4.6 mm×250 mm,5μm),流动相:乙腈-磷酸二氢钾缓冲溶液(30 mM)(10∶90,v/v),流速:1.0 mL/min,进样量:20μL,紫外检测波长:236 nm,柱温:20℃。结果本法在万古霉素浓度为5～200μg/mL浓度范围内线性关系良好(r=0.998 2,n=6),血浆最低检测浓度为5μg/mL,提取回收率在68%～81%;日内、日间RSD<5%,稳定性考察RSD<10%。结论本方法快速简便,灵敏度和准确度高,适用于万古霉素的人体药动学研究。

Objective To establish an HPLC method for the determination of plasma vancomycin concentra- tion, and to study its pharmacokinetics in healthy volunteers. Methods Taking vancomycin as external standard, after a extraction with 10% perchloric acid, the liquid chromatography was performed on Aglient ZORBAX SB-CI8 （4. 6 mm × 250 mm, 5 μm）column, with a mobile phase of acetonitrile and potassium dihydrogen phosphate buffer solution （30 mM,pH 3.2） （lO：90,v/v） ,and the flow rate was 1.0 mL/min. The injection volume was 20 μL,the UV detection wavelength was 236 nm and the column temperature was 20 ℃. Results The linear range of vancomycin was 5 - 200 μg/mL（r =0. 998 2,n =6）. The inspect limitation was 5 μg/mL,the extraction recovery was between 68% - 81% , the intra-day and inter-day RSD were less than 5% ,the stable inspection RSD was less than 10%. Conclusion The method developed in this study was proved to be rapid, simple, accurate and sensitive,it is suitable for the pharmacoki- netics study of vancomycin in people.