Development and Validation of RP-HPLC Method for Estimation of Winter green Oil in Pharmaceutical Formulation

Abstract

Background Wintergreen oil WO is used in various applications including aromatherapy and in topical pain relief products. The main active compound in wintergreen oil is methyl salicylate which is related to salicylic acid a component found in aspirin.Aim To develop and validate the RP-HPLC method for quantifying WO using methyl salicylate as the reference standard.Method In this study the pharmaceutical formulation chosen were nanosponges. A Gemini NX C18 250 mm X 4.6 mm 5 m column was used for the chromatographic separation carried out at room temperature with UV detection at 268 nm. A 1.5 mL min-1 flow rate was employed for the mobile phase which was a 5050 vv combination of acetonitrile and water. As per ICH requirements the approach was validated.Results The created technique showed R2 value of 0.9983 and exhibited linearity over 300-1500 microgmL concentration range. It was discovered that the limit of detection LOD and the limit of quantification LOQ levels were 0.082 microgmL and 0.25 microgmL respectively. The RSD values for repeatability intermediate and method precision were found to be within the limits of 2. The recovery ranged between 100.4 to 100.8 and negligible variations in RT and peak area with deliberate variations in pH flow rate wavelength and mobile phase ratio indicated robustness. The entrapment efficiency and loading capacity of nanosponges were determined to be 73.7-97.4 and 6.85-0.07 respectively.Conclusion The proposed method was proven to be particular straightforward sensitive precise and adaptable. As a result the technique was successfully utilized to estimate wintergreen oil in pharmaceutical formulations.