HPLC同时检测血浆中5-氟脲嘧啶和丝裂霉素的浓度

Simultaneous determination of fluorouracil and mitomycin C in plasma by HPLC

目的 建立高效液相色谱同时测定人血浆中化疗药物 5 氟脲嘧啶 (5 FU)和丝裂霉素 (MMC)浓度的方法。方法 以 5 氯脲嘧啶 (5 CU)和对氨基苯乙酮为内标 ,采用惠普 110 0型高效液相色谱仪 ,以HPHypersilODScolumn(12 5mm× 4mm ,5μm)为分析柱 ,前加HPLicrosphereC18保护柱 ,以乙腈和水为流动相低压梯度洗脱 ,紫外可变波长检测器在 2 5 4nm波长检测5 FU和 5 CU ,310nm检测对氨基苯乙酮 ,36 5nm检测MMC ,同时测定 5 FU和MMC的浓度。血浆样本用醋酸乙酯分二步直接萃取两种药物及其内标 ,40℃氮气吹干 ,残留物 10 0 μL水溶解后 ,2 0 μL进样。 结果 样品中各成分分离良好 ,无干扰。线性方程为 :5 FU :Y =116 2X - 3.16 2 ,r=1.0 0 0 0 ;MMC :Y =339.4X - 1.0 6 2 ,r=0 .9989。线性范围 :5 FU :5 0～ 2 5 0 0ng·mL-1;MMC :10～ 5 0 0ng·mL-1。检测限 :5 FU :10ng·mL-1;MMC :10ng·mL-1。提取回收率 :5 FU ,5 CU ,MMC约 70 % ;对氨基苯乙酮为 99%。结论 本法适用于包含 5 FU和MMC的化疗方案的监测及药动学的研究。

OBJECTIVE: To develop a high-performance liquid chromatography method for simultaneously determining the concentrations of fluorouracil (5-FU) and mitomycin C(MMC), the chemotherapeutic drugs in human plasma. METHODS: The HP 1100 HPLC system was used, including a quaternary pump, a manual injector, a variable wavelength detector and a LC 2D ChemStation. The analytical column was a HP Hypersil ODS column(125 min × 4 mm, 5 μm) with a HP Licrosphere C18 guard column. The gradient mobile phase, consisted of acetonitrile and water was flowed through the system with the speed of 0.8 ∼ 1.2 mL&middotmin-1. The gradient rate of acetonitrile and water was became from 0% to 28%. The detective UV wavelength was 254 nm for 5-FU and 5-CU (IS), 310 nm for 4-aminoacetophenone(IS) and 365 nm for MMC. The plasma samples, added 5-chlorouracil (5-CU) and 4 aminoacetophenone as the internal standards, were extracted two times with ethylacetate. The organic phase was evaporated by N2 at 40°C. The residue was with 100 μL water. 20 μL was injected. RESULTS: The chromatography was good and not interfered by the components of the plasma. The liner equations were Y= 1 162 x - 3.162, r = 1.0000 (50∼2 500 ng&middotmL-1) for 5FU and Y= 339. 4x - 1.062, r = 0.998 9(10∼500 ng&middotmL-1) for MMC, respectively. The RSD of intraday was less than 5%, and the RSD of interday was less than 10%. The extracted recovery of 5-FU, 5-CU and MMC was about 70%. CONCLUSION: It appeared to be an accurate, convenient and effective method which can simultaneously determine the concentrations of 5-FU and MMC in plasma.