LC-MS/MS method using unbonded silica column and aqueous/methanol mobile phase for the simultaneous quantification of...

BMS-754807 and metformin were co-administered in drug discovery studies which required the quantitation of both compounds in plasma. Since the two compounds are chemically and structurally dissimilar, developing a single bioanalytical method presented a number of chromatographic challenges including the achievement of appropriate retention times and peak shapes on a single analytical column. To address this chromatographic challenge, we investigated different LC columns under different gradient elution schemes using aqueous/organic mobile phases. Using unbonded silica column and aqueous/methanol mobile phase, we were able to obtain robust and well-resolving chromatographic conditions to support the development and implementation of a single LC-MS/MS bioanalytical method. The use of sub-2 micron particle sizes and a high flow rate, which are attainable with UPLC systems, enhanced the method. The method performance evaluation showed that the method easily met the normally used acceptance criteria for bioanalytical methods, namely a deviation of +/-15% from the nominal concentration except at lower limit of quantitation (LLOQ), where +/-20% is accepted. The reported LLOQ of 7.8 ng/ml, for both BMS-754807 and metformin, was adequate to support the pharmacokinetic studies.