高效液相色谱法测定人血浆中甲氨蝶呤浓度

Determination of Methotrexate in Human Plasma by HPLC

目的:建立高效液相色谱测定人血浆中甲氨蝶呤(MTX)浓度的方法。方法:以茶碱(Theophyline)为内标,色谱柱为Nova-pak C_(18)柱(150 mm×3.9 mm,4μm),流动相:乙腈-甲醇-0.05 mol·L^(-1)磷酸二氢钾缓冲液pH 6.5(6:2:92),流速:1.0 ml·min^(-1),检测波长303 nm及254 nm,柱温25℃。结果:甲氨蝶呤浓度在0.05～10.0 mg·L^(-1)范围内线性良好,r=0.999 6。绝对回收率为66.72%,相对回收率为98.35%,日内、日间RSD均<10%。结论:该法快速,简便,灵敏度高,适用于临床常规血药浓度监测。

Objective ： To establish an HPLC method for the determination of methotrexate （MTX） in human plasma. Method ： Theophyline was used as the internal standard. The analytical column was Nova-pak C18 column （3.9 mm×150 mm,4 μm）. The mobile phase was aeetonitrile-methanol-the buffer of 0.05 mol·L^-1 potassium dihydrogen phosphate with pH 6. 5 （2： 6： 92）. The UV detection wavelength was 303 nm and 254 nm and the column temperature was 25℃. Result： The calibration curve of methotrexate was linear within the range of 0. 05- 10. 00 mg·L＆-1, The intraday and inter-day RSD were both less than 10%. The absolute recovery and method recovery of MTX was 66. 72% and 98.35%, respectively. Conclusion： The method is simple, accurate and feasible to determine the concentration of methotrexae in human plasma.