Simultaneous determination of aliskiren and hydrochlorothiazide in tablets and spiked human urine by ion-pair liquid chromatography

摘要

An alternative method for analysis of aliskiren (ALI) and hydrochlorothiazde (HCT) in combined dosage forms by ion-pair reversed phase high performance liquid chromatography was developed and validated. The pharmaceutical preparations were analyzed using a C18 column (250mm x 4.6 mm, 3 μm) with a mobile phase consisting of 25% methanol, 50% sodium monobasic phosphate aqueous solution containing 6mMtetrabutylammonium bromide and 25%water at pH 7.2. Isocratic analysiswas performed at a flow rate of 1 mL/min and a column temperature of 30°C under direct UV detection at 210 nm. Paracetamol was used as internal standard. The validation was performed according to the ICH guidelines. The proposed method was linear over the concentration range of 0.250 to 60 and 0.1 to 10 μg/mL for ALI and HCT, respectively. The limits of detection and quantitation (LOD and LOQ) were 0.075 and 0.198 μg/mL, respectively, for ALI and 0.04 and 0.062 μg/mL, respectively, for HCT. The method proved to be specific, sensitive, precise and accurate with mean recovery values of 101.1 ± 0.32% and 100.9 ± 0.41% for ALI and HCT, respectively. The method robustness was evaluated by means of an experimental design. The proposed method was applied successfully to spiked human urine samples with mean recoveries of 98.8 ± 0.36% and 98.1± 0.21% for ALI and HCT, respectively.