建立人血浆中替加氟高效液相色谱测定法

 目的建立人血浆中替加氟(tegaful)高效液相色谱测定法。方法取0.2mL血浆,以甲硝唑(metronidazole)为内标,采用叔丁基甲醚与正丙醇的混合溶剂(80:20)提取后氮气吹干,复溶,进样分析。流动相:乙腈-水(25:200);流速1.0mL·min^-1;紫外检测波长270nln,室温操作。结果替加氟与内标分离良好,保留时间分别为10.54和9.31min,且达峰处无杂峰干扰。标准曲线范围0.10-25.0μg·mL^-1,线性良好(r=0.9994,n=5),批间误差0.29%-5.65%(n=5),批内误差1.29%-6.37%(n=10),最低检测限为0.02μg·mL^-1。替加氟血样在检测过程保持稳定。结论本方法取样量小,操作快速简便,专一性强,灵敏度高,可满足药动学研究需要。

Study on determination of tegafur in human plasma by HPLC assay

OBJECTIVE To develop a HPLC assay for the determination of tegafur in human plasma.METHODS 0.2 mL plasma sample with metronidazole as the internal standard was extracted with the mixture of methyl t-buryl ether and n-propylene(80:20) . The organic layer was evaporated to dry under nitrogen stream at room temperature. The residue was reconstituted with 100 μL mobile phase. The mobile phase consisted of acetonitrile-H₂O(25:200) at the flow rate of 1.0 mL·min^-1. The detection wavelength was at 270 nm.RESULTS The assay allowed excellent linearity(r=0.9994, n=5) over the range of 0.10-25.0 μg-mL^-1, and the limit of detection was 0.02 μg-mL^-1. The coefficient variations for 0.25,10.0 and 20.0 μg-mL^-1 check samples were 0.29% to5.65% for inter-day validation(n=5), and 1.29% to 6.37% for intra-day vahdation(n=10) , respectively. The average recoverys were 100.40% , 93.88% and 99.57% (n=10) , respectively. CONCLUSION The improved method was sensitive, specific, reproducible and easy to process. It is practical and suitable for the pharmacokinetic study of tegafur.