Validation of Reversed Phase Liquid Chromatographic Method for the Simultaneous Estimation of Irbesartan and Hydrochlorothiazide in their Pharmaceutical Preparations

Abstract

Background: The present study was aimed to develop and validate a maintenance of robust, accurate and reproducible isocratic reverse-phase high-performance liquid chromatography (RP-HPLC) method that permits the simultaneous quantitation of irbesartan (IRB) and hydrochlorothiazide (HCT) in composite tenders dosage forms. Methods: Column consisted of ODS C18 column (250 × 4.6 mm, 5 μm); the buffer-excluding mobile phase was departed by distilled water and methanol in a ratio of 50:50 (v/v). Analysis was carried out at 1.0 mL/min, and UV detection was detected using K260 Ap chromato-graphic system at275 nm. Were used a sample injection volume of 10 μL, and the total chromatographic run time was held at 10 min. Results: Chromatogram of IRB and HCT were obtained with good chromatographic resolution, resulting in retention times of 2.739 min and 3.558 min respectively for IRB and HCT (Fig. Our approach was found to be highly linear between 1 and 100 μg/mL for both analytes, with correlation coefficients (R²) of all standard curves being higher than 0.9994 or greater for IRB and by HCT equal to 0.9995. The data confirmed that the extraction methodology secured recovery values of 101.3232% and 100.0476%, respectively, of IRB and HCT, which was adequate for compliance with analytical validation requirements. Additionally, the method exhibited a high sensitivity with a limit of detection (LOD) and limit of quantification (LOQ) values equal to or less than 0.05 μg/mL and 0.165 μg/mL respectively. Conclusions: The validation results confirmed that the developed RP-HPLC method is precise, accurate, selective and robust for simultaneous quantitation of IRB and HCT. In addition, the method provides several practical benefits: simple chromatographic conditions ( ternary mixtures), a buffer-free mobile phase and comparatively short analysis time as well. These features allow that this method offers the reliability and economy required as a quality-control technique to analyze pharmaceutical formulations for of combined IRB and HCT tablets in the routine.