Micro-flow liquid chromatography coupled with a mass spectrometer (MFLC-MS/MS) has many advantages (sensitivity increase, less solvent consumption and reduced source contamination). These advantages have led to the manufacture of specialized LC pumps that accurately deliver ≤100 μL/min of solvent and LC columns with an inside diameter of ≤1 mm. In order to determine if the MFLC-MS/MS system can provide bioanalytical data that exceeds the criteria required by the FDA, a validation (determination of Methotrexate in human plasma) was performed. A validation was also performed using the conventional HPLC-MS/MS for comparison purposes.

Here we report data generated for the determination of Methotrexate from human plasma by MFLC-MS/MS and HPLC-MS/MS.