RP-HPLC法同时测定人体血清及脑脊液中万古霉素的含量

目的:建立人体血清及脑脊液样品中万古霉素浓度的高效液相色谱测定法。方法:以外标法定量,采用AglientZORBAXSB-C18色谱柱(4.6×250mm,5μm),流动相为30 mM磷酸二氢钾缓冲溶液-乙腈(91:9,pH3.2),流速1.1ml/min,检测波长236nm,柱温20℃。结果:万古霉素在5～200μg/mL血清浓度范围内线性关系良好(r=0.9981),最低定量限为5μg/mL,提取回收率在23.40～47.36%;日内RSD为3.57%、0.97%、0.46%(n=5),日间RSD为4.49%、0.90%、4.10%(n=5);在10～2000μg/mL脑脊液浓度范围内线性关系良好(r=0.9997),最低定量限为10μg/mL,提取回收率在48.61～53.20%;日内RSD为3.10%、6.34%、4.06%(n=5),日间RSD为19.47%、0.87%、2.53%(n=5)。结论:本方法快速,准确,具有创新性。

Determination of Vancomycin in Human Serum and Cerebrospinal Fluid by RP-HPLC

Objective： To establish an HPLC method to determine the concentration of vancomycin in human serum and cerebrospinal fluid.Method： Using the external standard method,the chromatography was preformed on an Aglient ZORBAX SB-C18（4.6×250mm,5μm） column with the mobile phase consisting of 30 mM potassium dihydrogen phosphate buffer solution-acetonitrile（91：9,pH3.2） with the flow rate 1.1 ml/min.UV detection was set to 236 nm and the column temperature was 20 ℃.Results： In the serum,the calibration curve was linear within the range 5～200μg/mL（r=0.9981）.The limit of test was 5μg/mL with the extraction recovery of 23.40～47.36%,and intra-day and inter-day RSD were 3.57%,0.97%,0.46%（n=5） and 4.49%,0.90%,4.10%（n=5）.In the cerebrospinal fluid,the calibration curve was linear within the range 10～2000μg/mL（r=0.9997）.The limit of test was 10μg/mL with the extraction recovery of 48.61～53.20%,and intra-day and inter-day RSD were 3.10%,6.34%,4.06%（n=5） and 19.47%,0.87%,2.53%（n=5）.Conclusion： The method is rapid,accurate and innovative.