Simultaneous analysis of betamethasone sodium phosphate and betamethasone dipropionate in the presence of degradation products by high pressure liquid chromatography

Steroids suppress proteins that mediate increased inflammation in cells. Corticosteroids are streoid hormones secreted from the adrenal cortex. Corticosteroids are widely used all over the world due to their anti-inflammatory effects. Bethamethasone is a corticosteroid that acts primarily as a glucocorticoid. In this study, high performance liquid chromatography method was developed and validated for the determination of betamethasone sodium phosphate, betamethasone dipropionate and preservativesin pharmaceutical preparations as injectable suspension. The proposed method was optimized by using different columns, mobile phase compositions and ion pair reagents. The analysis of these active substances and preservatives were performed on Zorbax XDB C18, 250 x 4.6 mm, 5 µm column, using composed of pH: 4.0 phosphate buffer : acetonitrile containing hexylamine as ion pair reagent by gradient elution, flow rate of 1.0 mL/min and wavelengths of 254 nm. In addition, stress test studies were also carried out to show the selectivity of the method. Thedeveloped analytical method was validated by applying the parameters of selectivity, linearity, accuracy, precision, ruggedness, and robustness. It has been suggested that the developed and validated analytical method can be used in the simultaneous analysis of betamethasone sodium phosphate, betamethasone dipropionate active substances and benzyl alcohol, methyl paraben and propyl paraben preservatives. This validated method was successfully applied to the pharmaceutical preparation containing these two active substances and preservatives. Keywords: Betamethasone sodium phosphate, Betamethasone dipropionate,Degradation products, High Performance Liquid Chromatography, Validation, Pharmaceutical preparation.