Determination of Ro 48-3656 in rat plasma by reversed-phase high-performance liquid chromatography: Comparison of 1.5-μm nonporous silica to 3.5-μm porous silica analytical columns

Abstract We describe a method for measuring Ro 48-3656 in EDTA rat plasma by neutral pH, reversed-phase high-performance liquid chromatography using a 1.5-μm nonporous silica, C 18 analytical column and UV absorbance detection to support pharmacokinetic studies. We also describe a comparison of the 1.5-μm nonporous silica C 18 column versus 3.5-μm porous silica C 18 columns. The final method using the 1.5-μm nonporous silica column demonstrated good precision (of both quantification and retention time), accuracy and recovery, linearity of dilution and limit of quantification (40 ng/ml Ro 48-3656 using a 20 μl injection). Samples of neat EDTA rat plasma were prepared by ultrafiltration followed by direct injection onto the HPLC column.