Development and Validation of a Stability-Indicating Liquid Chromatographic Method for Determination of Valsartan and Hydrochlorthiazide Using Quality by Design

The present paper involves the analytical quality by design (AQbD)-based development of a simple, rapid, accurate and precise stability-indicating method for the estimation of valsartan and hydrochlorothiazide. Optimized mobile phase (v/v/v) was water (containing 0.25 ml/L triethylamine), methanol and acetonitrile (50:38:37, pH adjusted to 3.0±0.1). Chromatographic separation was achieved on Hypersil®-Gold C18 (150 x 4.6 mm, 5 μm, Thermo Fisher Scientific, USA), column at 25 ± 2 °C. The method was validated as per the “International Conference on Harmonisation” (ICH) guidelines. Chromatographic run time was 10.0 minutes. The linearity range for valsartan and hydrochlorothiazide were 1.25-64.00 μg/ml and 0.195-10.00 μg/ml respectively. The limits of detection (LOD) for valsartan and hydrochlorothiazide were 0.253 and 0.0226 μg/ml while the limits of quantitations (LOQ) were 0.767 and 0.068 μg/ml respectively, using 10μl sample. Stability studies indicate that the degradation of valsartan was higher during oxidative stress than other stress conditions. keywords: Valsartan, hydrochlorothiazide, Central Composite Design, Design of experiments (DoE), Analytical Quality by Design (AQbD), Response surface methodology (RSM), HPLC.