Tabriz University of Medical Sciences Advanced Pharmaceutical Bulletin 2228-5881 3 2 2013 12 30 Development and Application of an HPLC Method for Erlotinib Protein Binding Studies 289 293 10.5681/apb.2013.047 EN Soheila Bolandnazar Adeleh Divsalar Hadi Valizadeh Arash Khodaei Parvin Zakeri-Milani Journal Article 10.5681/apb.2013.047 2013 02 10 Purpose: The aim of the present study was to develop a simple and rapid reversed-phase high performance liquid chromatographic method with UV detection for erlotinib hydrochloride quantification, which is applicable for protein binding studies. Methods: Ultrafilteration method was used for protein binding study of erlotinib hydrochloride. For sample analysis a simple and rapid reversed-phase high performance liquid chromatographic method with UV detection at 332 nm was developed. The mobile phase was a mixture of methanol, acetonitril and potassium dihydrogen phosphate buffer (15:45:40 %v/v) set at flow rate of 1.3 ml/min. Results: The run time for erlotinib hydrochloride was approximately 6 minutes. The calibration curve was linear over the range of 320-20000 ng/ml with acceptable intra- and inter-day precision and accuracy. The intra-day and inter-day precisions were less than 10% and the accuracies of intra and inter-day assays were within the range of 97.20-104.83% and 98.8-102.2% respectively. Conclusion: Based on the obtained results, a simple, accurate and precise reversed-phase isocratic HPLC method with UV detection has been optimized and validated for the determination of erlotinib hydrochloride in biological samples.