HPLC法测定人血清中甲氨蝶呤的浓度

目的:建立以高效液相色谱法测定人血清中甲氨蝶呤浓度的方法。方法:色谱柱为WatersC18,流动相为乙腈-pH7.2的磷酸盐缓冲液(12∶88),柱温为30℃,流速为1.0mL.min-1,检测波长为303nm。结果:甲氨蝶呤血药浓度在0.5～80μg.mL-1范围内与峰面积线性关系良好(r=0.9994);方法回收率均>90%,日内、日间RSD分别为3.03%～3.52%、2.57%～4.05%。结论:本方法简便、快速、准确、灵敏,适用于甲氨蝶呤体内血药浓度测定和药动学研究。

Determination of Methotrexate in Human Serum by HPLC

OBJECTIVE： To determinate the serum concentration of methotrexate by HPLC. METHODS： The samples were separated on a Waters C18 column at a column temperature of 30 ℃ . The mobile phase consisted of acetonitrile - phosphate buffer（pH 7.2, 12 ： 88） at a flow rate of 1.0 mL · min ^-1. The detection wavelength was set at 303 nm. RESULTS： The linear range of methotrexate was 0.5--80/μg · mL^-1（r = 0.999 4） . The methodological recovery rates were all above 90% ; the intraday RSD was 3.03 %-3.52 % and the interday RSD was 2.57%--4.05%. CONCLUSION：The method is proved to be simple, rapid, accurate and sensitive, and it is suitable for the determination of serum concentration and pharmaeokinetic study of methotrexate.