HPLC法测定人血浆中5-氟尿嘧啶的浓度

Determination of 5-fluorouracil Concentration in Human Plasma by HPLC

目的:建立测定人血浆中5-氟尿嘧啶(5-FU)浓度的方法。方法:血浆采用乙酸乙酯二次提取处理后以高效液相色谱法进样测定,其中,色谱柱为UltimateC18,流动相为10mmol·L-1KH2PO4溶液,流速为1.0mL·min-1,检测波长为265nm,柱温为40℃。结果:5-FU血药浓度在0.48～61.4μg·mL-1范围内线性关系良好(r=0.9998),最低检测浓度样品在为0.48μg·mL-1,日内和日间RSD分别为5.12%～9.98%、5.28%～10.95%(n=5),高、中、低3种浓度的方法回收率均高于85%。样品在室温放置、冷冻保存、反复冻融条件下稳定性良好。结论:该方法具有操作简便可靠、准确、稳定性高等特点,适用于人血浆中5-FU浓度的测定。

OBJECTIVE： To develop a method for the determination of 5-fluorouracil （5-FU） concentration in human plasma. METHODS： Plasma sample was treated with twice extraction of ethyl acetate, and then HPLC method was adopted for concentra- tion determination. The determination was performed on Ultimate C18 column with mobile phase consisted of 10 mmol·L-1 KH2PO4 at the flow rate of 1.0 mL·min-1. The detection wavelength was set at 265 nm and column temperature was 40 ℃. RESULTS： The linear range of 5-FU was 0.48-61.4 μg·mL-1（n=0.999 8）. The lowest limit of determination was 0.48 μg·mL-1. The RSD of in- tra-day and inter-day were 5.12%-9.98% and 5.28%-10.95% respectively （n=5）. The recovery rates were all higher than 85% at high, medium and low conentrations. The samples were stable in room and low temperature, and were also stable in frozen and thawed condition. CONCLUSION： This method is simple, precise and stable, and it is suitable for the determination of 5-FU con- centration in human plasma.