摘要
5-Fluorouracil (5-FU) has a broad spectrum of anti-tumor activity, widely applied to the treatment of cancers. However, it is necessary to determine the plasma concentration of 5-FU in clinical practice due to its narrow therapeutic index. Therefore, a simple, economic and sensitive high-performance liquid chromatography (HPLC) method was developed and validated for the determination of 5-FU in human plasma. Ethyl acetate was chosen as extraction reagent. Chromatographic separation was performed on a Diamonsil C18 column (250 mm × 4.6 mm i.d., 5 μm) with the mobile phase consisting of methanol and 20 mmol/L ammonium formate using a linear gradient elution at a flow rate of 0.8 mL/min. 5-FU and 5-bromouracil (5-BU) were detected by UV detector at 265 nm. The calibration curve was linear over the concentration range of 5—500 ng/mL and the correlation coefficient was not less than 0.992 6 for all calibration curves. The intra- and inter-day precisions were less than 10.5% and 4.3%, respectively, and the accuracy was within ±3.7%. The recovery at all concentration levels was 80.1±8.6%. 5-FU was stable under possible conditions of storing and handling. This method is proved applicable to therapeutic drug monitoring and pharmacokinetic studies of 5-FU in human.
5-Fluorouracil （5-FU） has a broad spectrum of anti-tumor activity, widely applied to the treatment of can- cers. However, it is necessary to determine the plasma concentration of 5-FU in clinical practice due to its narrow therapeutic index. Therefore, a simple, economic and sensitive high-performance liquid chromatography （HPLC） method was developed and validated for the determination of 5-FU in human plasma. Ethyl acetate was chosen as ex- traction reagent. Chromatographic separation was performed on a Diamonsil C18 column （250 mm × 4.6 mm i.d., 5 μm） with the mobile phase consisting of methanol and 20 mmol/L ammonium formate using a linear gradient elution at a flow rate of 0.8 mL/min. 5-FU and 5-bromouracil （5-BU） were detected by UV detector at 265 nm. The calibra- tion curve was linear over the concentration range of 5--500 ng/mL and the correlation coefficient was not less than 0.992 6 for all calibration curves. The intra- and inter-day precisions were less than 10.5%, and 4.3%, respectively, and the accuracy was within ＋3.7%. The recovery at all concentration levels was 80.1-8.6%. 5-FU was stable under possi- ble conditions of storing and handling. This method is proved applicable to therapeutic drug monitoring and pharma- cokinetic studies of 5-FU in human.
基金
Supported by National Natural Science Foundation of China (No. 30630075 and 20675056)
Major State Basic Research Development Program of China ("973" Program) (No. 2006CB933303)
