This application note describes a HPLC method for the analysis of methotrexate. The method uses a 10/90 acetonitrile/pH 6.0 phosphate/citrate buffer mobile phase and a ZORBAX Eclipse XDB-C18 column, column dimensions 3.9 x 300 mm, particle size 10 µm.
This document describes an application for High-Performance Liquid Chromatography (HPLC) analyzing Methotrexate according to United States Pharmacopeia (USP) standards. It highlights the performance of the ZORBAX Eclipse XDB-C18 column as a modern alternative to older L1 columns, specifically noting improvements in peak shape and efficiency.
| Column Particle Size | 3.5 and 5.0 um |
|---|---|
| Mobile Phase Ratio | 10:90 ACN : Buffer |
| Flow Rate (Original) | 2.0 ml/min |
| Column Dimensions (Modern) | 4.6 x 150 mm |
| Flow Rate (Modern) | 1 mL/min |
| Detection Wavelength | 302 nm |
The ZORBAX Eclipse XDB-C18 is available as a state of the art L1 alternative.
Benefits include improved peak shape, higher efficiency, greater sensitivity, reduced analysis time, and reduced back pressure.
Resolution is greater on the ZORBAX Eclipse XDB-C18 because the methotrexate peak is much sharper.