高效液相色谱法测定人血浆中卡培他滨浓度

Study on the determination of capecitabine in human plasma by HPLC

目的 :建立人血浆中卡培他滨 (capecitabine)高效液相色谱测定法。方法 :取 0 .3mL血浆 ,以甲硝唑为内标 ,叔丁基甲醚提取后氮气吹干 ,复溶 ,进样分析。流动相 :乙腈 水 (35∶6 5 ) ;流速 1 .0mL·min-1 ;紫外检测波长 31 0nm ,室温操作。结果 :卡培他滨与内标分离良好 ,保留时间分别为 6 .9min和 3.4min ,且达峰处无杂峰干扰。标准曲线范围 0 .0 5～ 2 .0 0mg·L-1 ,线性良好 (r=0 .9991 ,n =6 ) ,日间误差 2 .5 7%～ 5 .93%,日内误差 1 .99%～ 5 .96 %,最低检测限为 0 .0 1mg·L-1 。卡培他滨血样在检测过程中保持稳定。结论 :本方法取样量小 ,操作快速简便 ,专一性强 ,灵敏度高 ,可满足药物动力学研究需要。

OBJECTIVE An HPLC assay was developed for the determination of capecitabine in human plasma. METHODS 0.3 mL plasma sample with metronidazole as the internal standard was extracted with methyl t-butyl ether. The organic layer was evaporated to dryness under nitrogen stream at room temperature. The residue was reconstituted with 100 μL of mobile phase. The mobile phase consisted of acetonitrile: H 2O (35∶65) at a flow rate of 1.0 mL·min -1. The detection wavelength was at 310 nm. RESULTS The assay allowed excellent linearity(r= 0.999 1) over the range of 0.05- 2.00 mg·L -1, and the limit of detection was 0.01 mg·L -1. The coefficients of variation for 0.10, 1.00, 1.8 mg·L -1 check samples were 2.57% to 5.93% for intra-day validation(n=8),and 1.99% to 5.96% for inter-day validation(n=10),respectively. The average recovery for them were 96.9%, 100.5% and 102.2%(n=10),respectively. CONCLUSION The improved method is sensitive and specific,reproducible. It is suitable for the PK study of capecitabine.