固相萃取高效液相色谱法测定人血清中万古霉素浓度

目的建立固相萃取高效液相色谱法测定人血清中万古霉素浓度。方法血清样品经C18固相萃取小柱净化处理,以去甲万古霉素为内标。色谱柱:Hypersil ODS柱(250mm×4.6mm,5μm),流动相:乙腈:0.05mol/LKH2PO4(含0.1%三乙胺,磷酸调pH=3.5)(9:91),检测波长:236nm,流速:1.0mL/min,柱温:30℃,进样量:50μL。结果万古霉素在2.5～80.0μg/mL范围内线性关系良好(r=0.9992),最低检测浓度1.0μg/mL,方法回收率97.1%～104.2%,绝对回收率>70%,日内、日间RSD低于6.1%。结论本方法快速、灵敏、准确,可用于临床血清万古霉素的浓度监测。

HPLC determination of vancomycin in human serum by solid phase extraction

Objective To develop a SPE-HPLC method for the determination of vancomycin in human serum.Methods The serum samples were cleaned up using SPE-C18 cartridge,norvancomycin was used as internal standard.The separation was carried out on a Hypersil ODS column(250 mm×4.6 mm,5 μm).The mobile phase consisted of acetonitrile and 0.05 mol/L potassium dihydrogen phosphate buffer(containing 0.1% triethyamine,adjusted to pH 3.5 by phosphoric acid)(9:91).The detection wavelength was at 236 nm,the flow rate was 1.0 mL/min,the column temperature was 30 ℃,and the sample loading volume was 50 μL.Results The linear range of vancomycin was 2.5～80.0 μg/mL(r=0.999 2),the lowest limit of detection was 1.0 μg/mL.The recovery rate was 97.1%～104.2%,and the absolute recovery rate was more than 70%.The intra-day and inter-day RSD were less than 6.1%.Conclusion The method is rapid,sensitive and precise,it is suitable for the determination of vancomycin in human serum.