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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 the determination of methotrexate in serum. METHODS: Gold (4.6 mm × 150 mm, 5 μm) and methanol-phosphate buffer (19:81, pH 7.2) were used as mobile phase. The detection wavelength was set at 306 nm and the column temperature was 35 ℃. Protein, and then neutralized with sodium hydroxide, injection. Results: The recovery rates of serum samples were 90.2%, 94.4% and 101.2% respectively. The intra-and inter-day error RSD was <9.0%, the lowest detection concentration was 0.002μmol.L-1 and the low concentration was 0.01-1.38μmol.L-1 Good, r = 0.999 7; high concentration of 1.38 ~ 103.5μmol.L-1 good linear relationship r = 0.999 5. Conclusion: The method is simple, sensitive, linear range and accurate.