A New Bioanalytical Method Development & Validation for Simultaneous Estimation of Esomeprazole and Naproxen in Human Plasma by Using RP-HPLC

Abstract

Aim: A new reverse phase high performance liquid chromatography (RP-HPLC) method for the quantitative determination of Esomeprazole and Naproxen in human plasma was developed and validated as per US-FDA guidelines. Methodology: The drug was spiked in the plasma and extracted with mobile phase by precipitation method. The extracted analyte was injected into Symmetry C18 (4.6 x 150mm, 5μm, Make: XTerra) or equivalent, maintained at ambient temperature and effluent was monitored at 285nm. The mobile phase was composed of potassium dihydrogen phosphate and acetonitrile [HPLC Grade] in the ratio of 60:40. The pH of the potassium buffer was adjusted to 3.0 by using Ortho Phosphoric Acid. The flow rate was maintained at 1.0 mL/min. Results: The developed method shows high specificity for Esomeprazole and Naproxen. The calibration curve for Esomeprazole and Naproxen was linear from 1.0 to 6.0 ppm (r2= 0.999) and 25.0 to 150.0 ppm (r2= 0.999) respectively. The inter-day and intra-day precision was found to be within limits. The proposed method was adequate sensitivity, reproducibility, and specificity for the determination of esomeprazole and naproxen in plasma. The Lower limit of quantification (LLOQ) for the drug Esomeprazole and Naproxen were found to be 0.04µg/ml and 0.4µg/ml respectively. The average percent recovery for the drugs Esomeprazole and Naproxen were found to be 98.97-99.84 & 99.80-100.95 respectively and reproducibility was found to be satisfactory. Conclusion: The proposed method was accurate, and precise for the quantification of Esomeprazole and Naproxen in the plasma. The proposed can also be used for routine analysis in quality control. The method was validated for parameters like selectivity, sensitivity, precision, intermediate precision, accuracy, linearity, recovery & stability. This RP -HPLC method is suitable for determining the concentration of Esomeprazole and Naproxen in plasma and it can applied for routine analysis for determination of the Esomeprazole and Naproxen from dosage form during pharmacokinetic study.