摘要
目的:建立测定人血清中甲氨喋呤及其代谢产物7-羟基甲氨喋呤的浓度的LC-MS/MS法,并应用于临床治疗药物监测。方法:采用D3-甲氨蝶呤同位素内标,血清样品经含内标乙腈沉淀蛋白处理。HPLC色谱柱为Sepax GP-pHenyl,柱温35℃,流速0.3 ML·min^(-1),流动相为0.1%甲酸和乙腈,梯度洗脱。质谱检测方式为ESI正离子模式,MRM扫描,监测甲氨蝶呤m/z 455～308,7羟基甲氨蝶呤m/z 471～324,D3甲氨蝶呤(内标)m/z 458～311。结果:甲氨蝶呤浓度在5～4 000 ng·ml^(-1)范围内与峰面积线性关系良好(r=0.999 9),7-羟基甲氨蝶呤浓度在1～800 ng·ml^(-1)范围内与峰面积线性关系良好(r=0.999 7)。甲氨蝶呤及7-羟基甲氨蝶呤日内及日间RSD<9.0%。甲氨蝶呤平均回收率为94.34%～115.08%,7-羟基甲氨蝶呤平均回收率为96.41%～103.04%。结论:本方法简便、准确、快速,适用于甲氨喋呤及其代谢产物7-羟基甲氨喋呤的血药浓度测定。
Objective： To establish an LC-MS/MS method to determine methotrexate and its metabolite 7-hydroxy-methotrexate in human serum. Method： D3-methotrexate was used as the internal standard with acetonitrile as the serum precipitant. A Sepax GP-phenyl column was applied with 0.1% formic acid and acetonitrile as the mobile phase with gradient elution at a flow rate of 0.3 ml ·min^-1 at 35℃. The mass spectrum was at positive ion mode and MRM scan, detecting methotrexate at m/z 455-308, 7-hydroxy-metho- trexate at m/z 471-324 and D3-methotrexate at m/z 458-311. Result： The calibration curve of methotrexate was linear within the range of 5-4 000 ng·min^-1 （ r = 0. 999 9） while 7-hydroxy-methotrexate was within 1-800 ng ·min^-1 （ r = 0. 999 7 ）. The intra- and inter-day RSDs were both less than 9.0%. The average recovery of methotrexate was 94.34-115.08% with 96.41-103.04% for 7-hydroxy-methotrexate. Conclusion： This method is convenient, accurate and rapid, and suitable for the detection of methotrexate and 7-Hydroxy- methotrexate in human blood serum.
出处
《中国药师》
CAS
2012年第2期191-194,共4页
China Pharmacist
