Determination of Hydrochlorothiazide and Two Major Degradation Products by Stability Indicating High Performance Liquid Chromatography

Background: Hydrochlorothiazide (HCTZ) is a thiazide diuretic which comprises two sulfonamide groups. The literature is not clear regarding the identification of the chromatographic peaks of its two major related substances: chlorothiazide and 4-amino-6-chloro-1,3-benzenedisulfonamide (DSA). Methods: In the present study, a simple, sensitive, and selective HPLC method was developed and validated for the assay of HCTZ, Chlorothiazide and DSA. The method was carried out on a C18 column, maintained at 40ºC. The mobile phase was composed of monobasic potassium phosphate buffer 0.02M pH 3.0/acetonitrile/methanol (82:9:9, v/v/v), run at a flow rate of 1.0 mL/min, and UV detection at 270 nm. Results: All related compounds including processing impurities and degradants from stressed samples were well separated from each other. The performance of this method was validated in accordance to the ICH guidelines and included specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness. Conclusion: Based on the results, the HCTZ degradation pathway was proposed and the validated HPLC method was successfully applied to the quantitative analysis of HCTZ in pharmaceutical formulations, contributing to improve quality control, to assure therapeutic efficacy and to clarify the literature.