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目的:建立人血浆中吉西他滨的RP-HPLC测定方法。方法:取血浆样品500μl,以去氧氟尿苷为内标,用无水乙醚提取,取上清水浴挥干,150μl流动相溶解,取20μl进行HPLC测定。色谱柱为Agilent C_(18)柱(250 mm×4.6 mm,5μm),流动相为40mmol·L~(-1)醋酸铵水溶液-乙腈(97:3),流速1.0ml·min~(-1),检测波长268nm。结果:吉西他滨线性范围为20～640ng·ml~(-1),方法回收率为97.74%～105.15%,提取回收率为75.08%～77.17%,日内和日间RSD均小于9.5%。结论:该法简便,快速,准确,重现性好,符合生物样品的测定要求。
Objective: To establish a method for the determination of gemcitabine in human plasma by RP-HPLC. Methods: Take 500μl of plasma sample, take deoxyfluorouridine as internal standard, extract with anhydrous ether, dry it with supernatant, dissolve in 150μl of mobile phase and take 20μl for HPLC. The column was Agilent C 18 column (250 mm × 4.6 mm, 5 μm) with a mobile phase of 40 mmol·L -1 ammonium acetate (97: 3) and a flow rate of 1.0 ml · min -1 ), Detection wavelength 268nm. Results: The linear range of gemcitabine was 20 ~ 640ng · ml ~ (-1). The recoveries were 97.74% ~ 105.15%. The recoveries of gemcitabine were between 75.08% and 77.17%. The intraday and interday RSD were less than 9.5%. Conclusion: The method is simple, rapid, accurate, reproducible, in line with the determination of biological samples.