高效液相色谱法测定人脑脊液中万古霉素浓度及临床应用

Determination of Vancomycin in Human Cerebrospinal Fluids by High-performance Liquid Chromatography

目的:采用HPLC-DAD快速有效地测定人脑脊液中万古霉素浓度,为颅内感染患者鞘内注射调整剂量提供依据。方法:前处理采用乙腈:6%高氯酸(1∶1,v/v)沉淀萃取,采用Eclipse XDB C_(18)柱(250 mm×4.6 mm,5μm);流动相:A-pH3.2磷酸二氢钾缓冲液,B-乙腈,梯度洗脱;流速:1 mL·min^(-1);检测波长:236 nm;柱温:30℃;进样量:20μL;二极管阵列检测。内标法定量,去甲万古霉素为内标。结果:万古霉素低、中、高3个浓度(3.125、25、50 mg·L^(-1))方法的绝对回收率分别为95.4%、97.5%、100.7%,日内精密度RSD小于2.76%,日间精密度RSD小于5.24%,基质效应小于1.08%。结论:本方法准确简便,符合万古霉素药物浓度监测的要求。

Objective:To establish a high-performance liquid chromatography(HPLC)method for determining concentrations of vancomycin in cerebrospinal fluids.Methods:Norvancomycin was used as the internal standard.The chromatographic columnr was Eclipse XDB C_(18)(250 mm×4.6 mm,5μm).The mobile phase was ammonium biphosphate buffer(A)-methanol(B),gradient elution:0-20 min:98%-90%A;20-25 min:90%-90%A;25-25.1 min:90%-98%A;25.1-30 min:98%-98%A.The detecting wavelength of ultraviolet was 236 nm.Results:Standard curve was linear in the range of 1.5625-100 mg·L^(-1).As for norvancomycin,the average method recovery rates of three concentrations(3.125,25,50 mg·L^(-1))were 95.4%,97.5%and 100.7%respectively.The within-day relative standard deviation(RSD)was less than 2.76%,and the between-day RSD was less than 5.24%,matrix effect less than 1.08%.Conclusion:The HPLC method is simple,sensitive,accurate and precise which is suitable for the cerebrospinal fluids and the pharmacokinetic study of vancomycin.