反相高效液相色谱法测定人血浆中吉西他滨浓度

建立人血浆中吉西他滨的RP—HPLC测定方法。方法：取血浆样品500μ1，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20μ1进行HPLC测定。色谱柱为Agilent C18（250mm×4.6mm，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。

Determination the Concentration of Gemcitabine in Human Plasma by RP-HPLC

To develop a RP-HPLC method for determination the concentration of gemcitabine in human plasma. Method： Doxifluridine was used as the internal standard. Plasma were extracted by diethyl ether and determined by HPLC. Chromatography was performed on Agilent C18 column （250 mm ×4.6 mm, 5 μm）. The mobile phase consisted of 40 mmol·L^-1 ammonium acetate- acetonnitrile （97：3 ） at the flow rate of 1.0 ml·min^-1 and UV detection wavelength was 268 nm. Result： Calibration curves were linear over the concentration range of 20 -640 ng·ml^-1. The method recovery was 97.74% - 105.15% and the extraction recovery was 75. 08% -77.17%. The intra-assay and inter-assay variability values were less than 9.5%. Conclusion： The established HPLC method is simple, accurate, sensitive and can be applied to the determination of gemcitabine concentration in human plasma.