高效液相色谱法检测脑脊液中万古霉素药物浓度方法学建立

目的：建立一种有效、灵敏、准确的可用于检测人脑脊液内万古霉素浓度的方法学。方法：色谱柱：Diamonsil C₁₈（200 mm×4.6 mm，5 μm），流动相：乙腈-0.05 mol·L^-1磷酸二氢钠溶液（pH 3.6）（9：91），流速：0.8 mL·min^-1，紫外检测波长：236 nm，内标：去甲万古霉素，柱温：40℃，进样量：20 μL。结果：万古霉素峰与其他药物峰分离良好，不受脑脊液内源性物质干扰。在0.40~101.00 μg·mL^-1范围内万古霉素浓度和两者峰面积比值呈线性关系（r=0.999 6），最低定量限0.40 μg·mL^-1。万古霉素和内标的保留时间分别为13 min和10 min，日内和日间RSD均小于10%，低、中、高3个浓度提取回收率分别为60%、30.6%、28.8%。结论：此方法具有较高可行性，准确度高，适用于脑脊液中万古霉素浓度检测。

Establishment of the determination method of vancomycin in human cerebrospinal fluid by RP-HPLC

OBJECTIVE To establish an effective,rapid and acute method to determine vancomycin concentration in human cerebrospinal fluid.METHODS Chromatography was performed on Diamonsil C₁₈ column (200 mm×4.6 mm,5 μm) (Dikama Technologies),with acetonitrile-sodium dihydrogen phosphate (pH=3.6) (9:91) as the mobile phase.The flow rate was 0.8 mL·min^-1 and the detection wavelength was 236 nm.Norvancomycin was used as internal standard.The column temperature was 40℃.RESULTS The peak of vancomycin divided perfectly from other components,and was not interfered by endogenous ingredients in cerebrospinal fluid. The calibration curve was linear over the concentration range of 0.404-101 μg·mL^-1 (r=0.999 6) with the lower limit of quantitation (LLOQ) at 0.404 μg·mL^-1.The retention times of vancomycin and norvancomycin were 13 min and 10 min,respectively.Intra-and inter-day RSD values were lower than 10%. Recovery rates were 65.90%,73.85% and 76.71% at high,medium and low concentrations,respectively.CONCLUSION This HPLC method for determining vancomycin in cerebrospinal fluid is feasible,accurate,and appropriate for clinic test.