HPLC法同时测定人体血浆中氟尿嘧啶及其前体药替加氟的浓度

Simultaneous HPLC determination of 5-fluorouracil and its prodrug tegafur in human plasma

目的:建立同时测定氟尿嘧啶及其前体药替加氟在人体血浆中浓度的 HPLC 方法。方法:以阿昔洛韦为内标,血浆样品用硝酸银(10%)沉淀蛋白质,采用 Zorbax-ODS C_(18)分析柱(4.6 mm×250 mm,5μm),检测波长为265 nm,流动相:0.01 mol·L^(-1)磷酸盐(用2 mol·L^(-1)氢氧化钠溶液调节 pH 为8.0)-甲醇(97:3),流速1.0 mL·min^(-1),柱温为45℃。结果:氟尿嘧啶、替加氟分别在0.1～20μg·mL^(-1)(r=0.9998)和0.2～40μg·mL^(-1)(r=0.9998)浓度范围内线性关系良好,最低检测浓度分别为5和10 ng·mL^(-1),方法回收率分别为97.2%～101.3%和98.3%～102.6%,日内、日间 RSD 小于6%。结论:方法灵敏、快速、准确,并适用于临床上氟尿嘧啶及前体药替加氟浓度监测。

Objective：To establish an HPLC method for the simultaneous determination of 5 - fluorouracil and its prodrng tegafur in human plasma. Method：Proteins in plasma sample were precipitated by silver nitrate（ 10% ） after addition of the internal standard. After centrifugation, the supernatant was injected for analysis by HPLC. Separation was achieved on Zorbax -ODS C18（4. 6 mm×250 mm,5μm） column at 45℃ ,and detected at 265 nm. The mobile phase consisted of 0.01 mol·L^-1 sodium phosphate buffer （adjusted pH to 8.0 with 2 mol·L^-1 NaOH solution ） - methanol （97：3 ） at a flow rate of 1.0 mol·L^-1. Results：The linear ranges （n = 7 ） for determination of 5 - fluorouracil and tegafur were 0. 1 - 20μg·mL^-1（ r = 0. 9998 ） and 0. 2 - 40 μg·mL^-1 （ r =0. 9998 ）, respectively. The detection limits of the assay were 5,10 ng · mL^- 1, respectively. The method recoveries were 97.2% - 101.3% and 98.3% -102.6% ,respectively; intra- day RSD and inter- day RSD were less than 6%. Conclusion： This method is sensltive, convenient, accurate, and is ideally suited for the therapeutic monitoring of 5 - fluorouracil and its prodrug tegafur.