RP-HPLC法同时测定人体血清及脑脊液中万古霉素的含量

摘要

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.%　　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A g l i e n t ZORBAXSB-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.