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目的 :建立生物样品中甲氨喋呤 (MTX)浓度测定方法。方法 :固相萃取净化和富集样品 ,高效液相色谱法测定血清和脑脊液中MTX含量。色谱条件 :以C18柱为分析柱 ,流动相为甲醇 -乙腈 -磷酸盐缓冲液 (2∶7∶91) ,流速 0 8mL·min-1,柱温 2 5℃ ,检测波长 313nm。结果 :在 0 0 5～ 80 μg·mL-1范围内呈线性 ,回收率为 90 %以上 ,血清中MTX最低检测限为 10ng·mL-1,脑脊液中最低检测限为 5ng·mL-1。结论 :本法简便、快速、灵敏度高、特异性强 ,适合于生物样品中MTX监测及药物动力学研究。
Objective: To establish a method for the determination of methotrexate (MTX) in biological samples. Methods: Purification and enrichment of samples by solid-phase extraction and determination of MTX in serum and cerebrospinal fluid by high performance liquid chromatography. Chromatographic conditions: the C18 column as the analytical column, the mobile phase methanol-acetonitrile-phosphate buffer (2:7:91), the flow rate of 0 8mL · min-1, column temperature 25 ℃, detection wavelength 313nm. Results: The calibration curve was linear over the range of 0 0 ~ 80 μg · mL-1 and the recovery was over 90%. The detection limit of MTX in serum was 10 ng · mL-1, and the detection limit of CSF in cerebrospinal fluid was 5 ng · mL-1. Conclusion: The method is simple, rapid, sensitive and specific. It is suitable for MTX monitoring and pharmacokinetic studies in biological samples.