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目的:建立以高效液相色谱法测定人血清中甲氨蝶呤(MTX)浓度的方法。方法:采用Shim-pack VP-ODS(150mm×4.6mm,5μm)的色谱柱;流动相:乙腈-0.3%磷酸(15∶85,用三乙胺调pH2.5);紫外检测波长为302nm;柱温:40℃;流速:0.8mL.min-1。结果:MTX在0.1~100.0mg.L-1质量浓度内线性关系良好,Y=0.2+30 174X,r=0.999 4;MTX低、中、高(0.25,12.50,100.00mg.L-1)3种质量浓度的平均回收率分别为104.0%,102.3%,102.5%。日内、日间RSD均小于3%,最低血清检测浓度50μg.L-1。结论:高效液相色谱法测定MTX的血药浓度灵敏度高,结果准确可靠,操作简便,能满足其血清浓度的测定。
Objective: To establish a method for the determination of methotrexate (MTX) in human serum by high performance liquid chromatography. Methods: The chromatographic column was Shim-pack VP-ODS (150mm × 4.6mm, 5μm). The mobile phase consisted of acetonitrile-0.3% phosphoric acid (15:85, adjusted to pH2.5 with triethylamine) Column temperature: 40 ° C; flow rate: 0.8 mL.min-1. Results: MTX showed a good linearity in the range of 0.1-100.0 mg.L-1 with Y = 0.2 + 30 174X, r = 0.999 4; MTX low, medium and high (0.25,12.50,100.00 mg.L-1) 3 The average recoveries of species concentration were 104.0%, 102.3% and 102.5%, respectively. Days, day RSD were less than 3%, the lowest serum test concentration 50μg.L-1. Conclusion: The HPLC method for the determination of MTX in plasma has high sensitivity, the results are accurate and reliable, easy to operate, and can meet the determination of serum concentration.