人血清中氟尿苷的HPLC测定方法

High performance liquid chromatographic determination of floxuridine in human serum

目的 建立人血清中氟尿苷的HPLC测定方法。方法 取血清 2 0 0 μL ,以甲硝唑为内标 ,采用正丙醇 /叔丁基甲醚作为提取液进行两次提取。常温氮气下吹干 ,流动相复溶后进样分析。色谱柱 :Shim PackCLC ODS柱 ;流动相 :乙腈 磷酸缓冲液 水 (75∶10 0∶90 0 ) ;流速 :0 .6mL·min-1;检测波长 :2 6 8nm。结果 在 0 .0 0 5～ 0 .5mg·L-1的范围内线性良好 ,相关系数r=0 .9999(n =8)。低、中、高 3个浓度质控样品的批内RSD在 1.97%～ 4.0 9%之间 ,批间RSD在 3.19%～ 3.96 %之间 ,方法学回收率为10 0 .88%～ 10 7.0 0 % ,富集进样时 0 .0 0 1mg·L-1的氟尿苷血清标准品可被检出。结论 本方法灵敏度高 ,操作简便易行 ,为研究其临床药物动力学。

OBJECTIVE To develop an HPLC method for the determination of floxuridine in human serum.METHODS With metronidazole as the internal standard,200 μL of serum was extracted by n propyl alcohol/methyl t butyl ether in a two step extraction.The organic layer was evaporated under nitrogen stream and the residual was reconstituted with the mobile plase.A Shim Pack CLC ODS column was selected and the mobile phase was consisted of acetonitrile phosphate buffer water (75∶100∶900) at a flow rate of 0.6 mL·min -1 .The detection wavelength was 268 nm.RESULTS A linearitywas obtained from 0.005 to 0.5 mg·L -1 of floxuridine in serum with a good correlation coefficient ( r =0.9999, n =8).The intra run and inter run coefficients of variation were less than 4.09%.The mean recoveries were 103.00%,107.00% and 100.88% for the low,middle and high concentrations of check samples,respectively.The limit of detection was 0.001 mg·L -1 .CONCLUSION The method was sensitive,specific and simple.It is suitable for clinical pharmacokinetic study.