Simultaneous Determination of Hydrochlorothiazide and Benazepril Hydrochloride or Amiloride Hydrochloride in Presence of Hydrochlorothiazide Impurities: Chlorothiazide and Salamide by HPTLC Method

Simple, selective and sensitive high-performance thin layer chromatographic (HPTLC) method has been developed and validated for the simultaneous determination of hydrochlorothiazide (HCZ) in the presence of its impurities (chlorothiazide (CT) and salamide (DSA)), in two quaternary mixtures with benazepril hydrochloride (BZ) or amiloride hydrochloride (AM). The separation was carried out on HPTLC silica gel 60 F-254 using ethyl acetate-methanol-glacial acetic acid (85: 2: 0.3 v/v/v) followed by densitometric measurement of bands at 240 nm for the first mixture containing HCZ, CT, DSA, BZ and by using ethyl acetate-methanol-water-ammonia (90: 10: 5: 3 v/v/v) followed by densitometric measurement at 278 nm for the second mixture containing HCZ, CT, DSA, AM. Calibration curves were constructed in the range of (0.2-1.8 mg/band) and (0.4-2.2 mg/band) with good accuracy for HCZ and BZ, respectively, for the first mixture and in the range of (0.6-1.8 mu g/band) and (0.4-2.4 mu g/band) with good accuracy for HCZ and AM, respectively, for the second mixture. The developed method was validated according to ICH guidelines and demonstrated good accuracy and precision. Moreover, the methods were successfully applied for the determination of HCZ and BZ and AM in pure form and pharmaceutical dosage forms. The results were statically compared with the reported methods with no significant difference, indicating the ability of the proposed method to be used for routine analysis of drug product.