Separation and quantification of organic‐related impurities of anti‐histamine drug hydroxyzine in pharmaceutical dosage forms using stability‐indicating high‐performance liquid chromatography, liquid chromatography‐mass spectrometry, and high‐resolution mass spectrometry techniques

Abstract

A simple and robust high‐performance liquid chromatography (HPLC) method was developed for organic impurities of hydroxyzine hydrochloride in pharmaceuticals. The developed method was designed to estimate all organic impurities of hydroxyzine. The HPLC separation was achieved using C18 column (150 × 3.9 mm, 5 μm) along with a binary gradient consisting of mobile phases A (0.1%, trifluoroacetic acid in purified water) and B (0.05%, trifluoroacetic acid in acetonitrile), a flow rate of 0.7–mL/min, a column temperature of 30°C and a sample temperature of 25°C. The detection wavelength used was 230 nm for the estimation of impurity‐A, impurity‐B, and all unspecified impurities and degradation products, whereas impurity‐C was quantitated using 254 nm. The stability‐indicating property of the developed HPLC technique was assessed using stress testing conditions of hydrolysis, oxidation, thermal, photo‐light, and humidity. The validation study was performed for the limit of detection and limit of quantification, linearity, and recoveries were 0.03%, 0.05%, and 0.1132–2.9920 μg/mL (R2 > 0.999), and 84.09%–109.74%, respectively. The proposed method is highly suitable for the determination of assay, organic impurities, and degradation products of the hydroxyzine. The chemical structure of degradation product 1 (hydroxyzine N‐Oxide) and degradation product 2 (O‐Acetyl hydroxyzine) were identified with the supporting data of LC‐mass spectrometry (LC‐MS) and high‐resolution MS.