关键词: 万古霉素, 超高效液相色谱, 治疗药物监测

Abstract:

Objective To establish ultra performance liquid chromatography（UPLC） for the rapid determination of vancomycin concentration, and to evaluate the performance. Methods Norvancomycin was used as internal standard,serum sample was treated with 0.3 mol/L aqueous solution of zinc sulfate,and a simple protein precipitation method was used for pre-treating serum samples. The chromatography was carried out on Phenomenex Kinetex C18 column（2.6 μm,100 mm×2.1 mm）,the flow rate was 0.5 mL/min,UV detector wavelength was 220 nm,and column temperature was 40 ℃. Using mobile phase acetonitrile-25 mmol/L potassium dihydrogen phosphate buffer（pH value was 2.5）,gradient elution was performed. The linearity,accuracy（recovery rate）,precision,carry-over rate,limit of quantitation and limit of detection were evaluated. A total of 50 clinical samples were determined by the present UPLC method and the validated high performance liquid chromatography（HPLC） method,and the results were compared using linear regression analysis. Results The linear range for serum concentration of vancomycin was 2.0-99.6 μg/mL,with 0.1 μg/mL detection limit and 1.0 μg/mL quantitation limit. The within-run coefficients of variation（CV）of low-,middle- and high-levels were 3.28%,2.21% and 2.59%,and the between-run CV were 5.73%,2.75% and 0.82%,respectively. The recovery rates of low-level（2.0 μg/mL）,middle-level（19.9 μg/mL） and high-level（79.6 μg/mL） were 104.06%,99.80% and 100.19%,respectively. The carry-over rate of middle- and high-levels were 0.57% and 0.19%,respectively. There was a correlation between the 2 methods（r=0.991 9）. Conclusions The established UPLC for the determination of vancomycin concentration is reproducible,simple,sensitive and accurate,which is suitable for monitoring clinic serum vancomycin concentration and experimental subject research.

Key words: Vancomycin, Ultra performance liquid chromatography, Therapeutic drug monitoring