HPLC法测定人血浆中万古霉素浓度的不确定度评定

Uncertainty evaluation on the determination of vancomycinin human plasma by HPLC method

目的评定液相色谱(HPLC)法测定人血浆中万古霉素(VCM)浓度的不确定度。方法采用Aglient ZORBAX SB-C18色谱柱(4.6 mm×250 mm,5μm),流动相为30 mmol/L磷酸二氢钾缓冲溶液-乙腈(91.5∶8.5 p H值3.38),流速1.1 m L/min,检测波长236 nm,柱温35℃。分析并评定HPLC法测定人血浆中VCM浓度过程中的不确定度来源并进行合成。结果人血浆中VCM低浓度(2.48μg/m L)和高浓度(172.2μg/m L)的扩展不确定度分别为0.168μg/m L和9.64μg/m L(P=95%,k=2)。结论用HPLC法测定人体血浆中万古霉素浓度的不确定度在高浓度时主要由基质效应、生物样品的配制及重复性引入,在低浓度时主要由回收率、基质效应及生物样品配制引入。

Objective To evaluate the uncertainty on the determination of vancomycin（VCM） in human plasma by HPLC method. Methods The chromatography was performed on an Aglient ZORBAX SB-C18（4.6 mm×250 mm, 5 μm） column with the mobile phase consisting of 30 mmol/L potassiumdihydrogen phosphate-acetonitrile（91.5∶8.5, p H 3.38） at the flow rate of 1.1 m L/min. UV detection was set at 236 nm and the column temperature was set at 35℃. The uncertainty sources in the determinationof VCM were analyzed and the uncertainty was evaluated and combined. Results The expanded uncertainty of VCM at low（2.48 μg/m L） and high（172.2 μg/m L） concentrations were 0.168 μg/m L and 9.64 μg/m L respectively（P = 95%, k = 2）. Conclusion The uncertainty on the determination of VCM in human plasma by HPLC method is mainly caused by recovery, matrix effectand sample preparation for low concentrations, and the matrix effect,sample preparation and repeatability for high concentrations.