HPLC测定大剂量甲氨蝶呤患者血药浓度

Determination of Methotrexate Level in Plasma Using Higher Doses Methotrexate by HPLC

目的:建立HPLC测定血清中甲氨蝶呤浓度的方法。方法:采用Gold色谱柱(4.6 mm×150 mm,5μm),流动相为甲醇-磷酸盐缓冲液(19∶81,pH 7.2),检测波长306 nm,柱温35℃,样品经高氯酸沉淀蛋白后,再用氢氧化钠中和,进样。结果:血清样品回收率分别为90.2%,94.4%,101.2%,日内和日间误差RSD<9.0%,最低检测浓度为0.002μmol.L-1,低浓度在0.01～1.38μmol.L-1线性良好,r=0.999 7;高浓度在1.38～103.5μmol.L-1线性关系良好r=0.999 5。结论:本方法简便、灵敏、线性范围宽、准确。

Objective：To establish a HPLC method for determination of methotrexate concentration in plasma using higher doses.Method：The separation was performed in a Gold column（4.6 mm × 150 mm,5 μm） with a mobile phase of methanol：phosphate buffer（19 ∶ 81,pH 7.2） and determinated at 306 nm.Result：The measurable lowest concentration was 0.002 μmol.L-1.The experiment showed that the recovery from plasma was 90.2%,94.4%,101.2%.The RSD of within day and between day was less than 9.0%.A good linearity was found in the range of 0.01 ～ 1.38 μmol.L-1（r = 0.999 7） and in the range of 1.38 ～ 103.5 μmol.L-1（r = 0.999 5）.Conclusion：The method is simple,sensitive and accurate with wide linear dynamic range.