Development and Validation of RP-HPLC Method for Simultaneous Estimation of Tramadol Hydrochloride and Diclofenac Sodium in Synthetic Mixture

摘要

A simple, accurate, and rapid reversed phase high performance liquid chromatographic method has been developed and validated for the simultaneous estimation of Tramadol Hydrochloride (TRM) and Diclofenac sodium (DIC) in synthetic mixture. The separation was carried out using mobile phase consisting of methanol: phosphate buffer (pH, 6.0) (68:32, v/v). The column used was Phenomenex C18, (250 mm x 4.6 mm i.d, 5 μm) with flow rate 1.2 ml/min using PDA detection at 219 nm. The method was linear over a concentration range 2-30 μg/ml for both drugs. The retention time of Tramadol hydrochloride and Diclofenac sodium were found to be 6.30 min and 8.35 min respectively. Results of analysis were validated statistically and by recovery studies. The mean recovery was 99.29 ± 0.54 and 99.03 ± 0.45 for TRM and DIC respectively. The limit of detection (LOD) and the limit of quantification (LOQ),for TRM and DIC were found to be 0.6586 and 1.9956 μg/ml and 0.6169 and 1.8695 μg/ml, respectively. The results of the study showed that the proposed RP-HPLC method was found to be simple, sensitive, precise and accurate and also useful for the routine determination of TRM and DIC in mixture.