高效液相色谱法测定人血浆中5-Fu浓度

摘要

Objective To develop a method for the determination of 5-fluorouracil concentration in human plasma.Methods 5-Bro-mouracil was used as the internal standard,ammonium sulfate was added to precipitate protein,plasma sample was treated with extrac-tion of ethyl acetate,and then HPLC method was adopted for concentration determination.The determination was performed on Hypersil ODS2 (5 μm,4.6 mm ×250 mm)column with mobile phase consisting of methyl alcohol-water (5∶95,v/v)at the flow rate of 0.8 mL ·min-1 .The detection wavelength was set at 266 nm and column temperature was 25℃.Results The linear range of 5-fluorouracil was 0.020~0.250 mg·L-1 (r=0.999 6).The lowest limit of quantification was 0.020 mg·L-1 .The RSD of intra-day and inter-day were 2.97%~11.17% and 1.67%~5.47%respectively.The recovery rates were all higher than 96%at high,medium and low con-centrations.The extraction rates were all higher than 70% at high,medium and low concentrations.The plasma samples were stable within 24 hours at room temperature and within 20 days in frozen condition.Conclusions This method is simple,precise and stable, which is suitable for the determination of 5-fluorouracil concentration in human plasma.%目的：建立测定人血浆中5-氟尿嘧啶（5-Fu）浓度的方法。方法以5-溴尿嘧啶（5-Bru）为内标，硫酸铵为蛋白沉淀剂，血浆采用乙酸乙酯提取处理后以高效液相色谱法进样测定，其中，色谱柱为Hypersil ODS2（5μm，4．6 mm ×250 mm），流动相为甲醇∶水（5∶95，V/V）溶液，流速0．8 mL·min-1，检测波长为266 nm，柱温为25℃。结果在0．020～0．250 mg·L-1浓度范围内药物浓度与响应值之间线性关系良（r＝0．9996），最低定量限浓度为0．020 mg·L-1，日内和日间RSD分别为2．97％～11．17％、1．67％～5．47％；高、中、低浓度的方法回收率均高于96％，高、中、低浓度的提取回收率均高于70％。血浆样品在室温24 h内及冷冻条件下20 d内稳定性良好。结论该方法具有操作简便可靠、准确、稳定性高等特点，适用于氟尿嘧啶植入剂人血浆中5-Fu浓度的测定。