论文部分内容阅读
目的建立灵敏、特异、快速、稳定可靠的甲氨蝶呤高效液相色谱检测方法,对患者血浆中甲氨蝶呤的血药浓度进行监测。方法色谱柱:SB-C18柱(4.6 mm×150 mm,5μm);流动相:甲醇-0.05mol·L~(-1)磷酸二氢钾缓冲液=26∶74(p H 2.4);流速:1.0 m L·min~(-1);检测波长:306nm;柱温:室温;进样量:40μL;甲硝唑为内标。结果血浆中甲氨蝶呤在0.04~4.0μg·m L~(-1)线性相关程度良好(R~2均大于0.99);低、中、高质量浓度质控样品(0.1,0.5,3.0μg·m L~(-1))的日内相对标准偏差(RSD)为2.12%~3.93%,日间RSD为3.48%~6.16%,绝对回收率在60.63%~75.72%。结论本研究建立的方法有良好的特异性、灵敏度、精密度和准确度,适用于临床患者甲氨蝶呤的血药浓度测定。
Objective To establish a sensitive, specific, rapid, stable and reliable method for methotrexate high performance liquid chromatography to monitor plasma concentrations of methotrexate in patients. Method Column: SB-C18 column (4.6 mm × 150 mm, 5 μm); mobile phase: methanol -0.05 mol·L -1 potassium dihydrogen phosphate buffer = 26:74 (p H 2.4); flow rate: 1.0 m L · min -1; detection wavelength: 306 nm; column temperature: room temperature; injection volume: 40 μL; metronidazole as internal standard. Results The linear correlation between methotrexate and plasma concentration was 0.04 ~ 4.0μg · m L -1 (R ~ 2> 0.99). Low, medium and high quality control samples (0.1, 0.5 and 3.0μg · M L -1) was 2.12% -3.93%, daytime RSD was 3.48% ~ 6.16%, and the absolute recovery was 60.63% ~ 75.72%. Conclusion The method established in this study has good specificity, sensitivity, precision and accuracy and is suitable for the determination of methotrexate in clinical patients.