Hossein Danafar - school of pharmacy zanjan university

Background and aims: A selective and sensitive high performance liquid chromatography-electrospray ionization mass spectrometry method has been established for determination of methotrexate in human plasma. Methods: Methotrexate was extracted from plasma with acetonitrile. The mobile phase consisted of acetonitrile-water-formic acid ۷۴: ۲۵: ۱(v/v). Twenty µl of sample was chromatographically analyzed using a repacked ZORBAX-XDB-ODS C۱۸ column (۲.۱mm×۳۰mm, ۳.۵ micron). The mode of mass spectrometry was selected-ion monitoring (SIM). Results: Standard curve was linear (r= ۰.۹۹۸) over the concentration range of ۰.۱–۱۰۰.۰ ng/ml and showed a suitable accuracy and precision. The limit of detection (LOD) was ۰.۰۵ng/ml. Conclusion: The method is quick, easy, very steady and precise for the partition, assignment and evaluation of methotrexate in human plasma.

Methotrexate, LC-MS, Human plasma