DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF TERBUTALINE SULPHATE, GUAIPHENESIN AND AMBROXOL HYDROCHLORIDE FROM AN ORAL LIQUID

A reverse phase high performance liquid chromatography method was developed and validated for simultaneous quantitative estimation of ambroxol hydrochloride, guaiphenesin and terbutaline sulphate. The chromatographic separation was achieved using a Phenomenex Luna C18 (2), 100A, 5μ (4.6 X 250mm) column using 0.012(M) sodium salt solution of 1-hexane sulphonic acid:acetonitrile 73: 27 (V/V) adjusted to pH 3 using orthophosphoric acid having isocratic elution of flow rate 1.0mL/min, detection at 220nm and injection volume 20 µL. The retention times of ambroxol hydrochloride, guaiphenesin and terbutaline sulphate were 25, 6.7 and 4.7 min, respectively. The proposed method was found to have linearity in concentration range of 63.40-95.11 μg/mL for ambroxol, 68.00-302.52 μg/mL for guaiphenesin and 4.88-7.20 μg/mL for terbutaline sulphate. Regression coefficient (R2) were 0.9997, 0.9989 and 0.9984 and percentage recovery were 101.67%, 101.68%, and 99.45% for ambroxol, guaiphenesin and terbutaline sulphate, respectively. The method validation showed excellent results for precision, accuracy, linearity and specificity. The proposed method was successfully used for the quantitative analysis of an oral liquid formulation, being developed in our laboratory, containing ambroxol hydrochloride, guaiphenesin and terbutaline sulphate.