Quantification of Tramadol hydrochloride and its Related Substances by HPLC in Pharmaceutical Dosage Form

A specific, linear, precise, accurate, robust and rugged reverse phase high performance liquid chromatography method is developed and validated for the estimation of Tramadol Hydrochloride in its capsule dosage form. The chromatographic separation is achieved on a Thermo, Hypersil BDS C8 column (250mm X 4.6mm, 5μm) with UV detection at 270nm. The optimized mobile phase consisted of acetonitrile and 0.2% v/v trifluoro acetic acid buffer (29.5:70.5% V/V) pumped at a flow rate of 1mL/min. Tramadol Hydrochloride is eluted at 6.36min (retention time). Based on the obtained validation results, the method is found to be specific, linear, precise, accurate, robust and rugged for Tramadol hydrochloride and it’s Impurity-A. The limit of detection and quantitation of Tramadol Hydrochloride are found to be 0.80μg/mL and 2.42μg/mL respectively. The limit of detection and quantitation of Impurity-A are found to be 0.51μg/mL and 1.54μg/mL respectively. The stability and filter integrity tests were performed and are found to be passed for Tramadol hydrochloride standard and sample solution. The proposed method can be successfully applied for the estimation of Tramadol Hydrochloride and its related compounds in its capsule dosage form.