A Novel Validated RP-HPLC Method For the Estimation of Salmeterol By DOE

Abstract

Background: Salmeterol is a long-acting beta agonist (LABAs) Salmeterol is hypothesized to bind to 2 sites on the beta-2 adrenoceptor. The saligenin moiety binds to the active site of the beta-2 adrenoceptor. The hydrophilic tail of salmeterol binds to leucine residues in the exo-site of the beta-2 adrenoceptor almost irreversibly, allowing salmeterol to persist in the active site, which is responsible for its long duration of action.

 Method: A new, simple, accurate, rapid, precise, reproducible and cost-effective RP-HPLC method for the quantitative estimation of Salmeterol in bulk and pharmaceutical dosage form. The developed RP-HPLC method for the quantitative estimation of Salmeterol is based on measurement of absorption at maximum wavelength 254 nm using Phosphate buffer: Methanol P^H 2.5 (35:65 v/v) as a solvent. The stock solution of Salmeterol was prepared, and subsequent suitable dilution was prepared in mobile phase to obtained standard curve. The standard solution shows absorption maxima at 254 nm.

Results: The Salmeterol obeyed Beer Lambert’s law in the concentration range of 20-100μg/ml with regression 0.999 at 254 nm. The overall % recovery was found to be 99.78% for Salmeterol which reflects that the method was free from the interference of the impurities and other excipients, used in the bulk and marketed dosage form. The low value of % RSD was indicative of accuracy and reproducibility of the method. The % RSD for inter-day and intra-day precision was found to be 0.2 for Salmeterol respectively which is <2% hence proved that method is precise.

Conclusion: In the present study, novel reverse phase High performance liquid chromatography method for simultaneous determination of Salmeterol in pharmaceutical dosage form was developed. The developed method was validated for various parameters such as accuracy, precision, ruggedness, linearity, robustness, system suitability, specificity as per ICH guidelines.