DOI: 10.21276/ajptr
Mon, 20 May 2019

Validated Stability Indicating RP-HPLC Method for Simultaneous Quantitative Estimation of Hydrochlorothiazide and Nebivolol Hydrochloride in Bulk and Combined Tablet Dosage Form

Mohammad Yunoos1*, D. Gowri Sankar2

1. Department of Pharmaceutical Analysis, Bapatla College of Pharmacy, Bapatla-522101, Guntur (Dist.), Andhra Pradesh, India.

2. College of Pharmaceutical Sciences, Andhra University, Visakhapatnam-530 003.


A simple and precise stability indicating RP-HPLC method was developed and validated for the simultaneous determination of Hydrochlorothiazide (HCTZ) and Nebivolol Hydrochloride (NBV) inbulk and Pharmaceutical dosage forms. Chromatography was carried out on Thermo Hypersil BDS C 18 (150 x 4.6 mm, 5mparticle size) column using a mobile phase of phosphate buffer (adjusted to pH 6.5 with dilute orthophosphoric acid): acetonitrile (40:60% v/v) at a flow rate of 0.8 ml/min. The analyte was monitored using PDA detector at 282 nm. The retention time was found to be2.4 min and 4.0min for Hydrochlorothiazide and Nebivolol Hydrochloride respectively. The proposed method was found to be having linearity in the concentration range of 6.25-37.5 µg/ml for Hydrochlorothiazide (r2 0.9999) and 2.5-15 µg/ml for Nebivolol Hydrochloride (r2 0.9999) respectively. The mean % recoveries obtained were found to be 99.93 % for Hydrochlorothiazide and 100.03% for Nebivolol Hydrochloride respectively. Stress testing which covered acid, alkali, peroxide, photolytic and thermal degradation was performed on under test to prove the specificity of the method and the degradation was achieved. The developed method has been statistically validated according to ICH guide lines and found to be simple, precise and accurate with the prescribed values. Thus the proposed method was successfully applied for the stability indicating simultaneous determination of Hydrochlorothiazide and Nebivolol Hydrochloridein bulk and Pharmaceutical formulations and in routine quality control analysis.

Keywords: Hydrochlorothiazide, Nebivolol Hydrochloride, RP-HPLC, Forced degradation, Method validation.

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