Method Development and validation by Liquid chromatography Mass spectroscopy (LC-MS) for Bilastine by a stability indicating assay method

Abstract

Bilastine, a newer second-generation antihistamine (SGAH) with minimal sedative effects, was recently introduced in the Indian market following approval from the Drugs Controller General of India. It has been prescribed for use in adults and adolescents aged 12 years and older across most countries since 2019. The recommended dosage is 20mg once daily for various treatment of allergic rhino-conjunctivitis and urticaria. Upon conducting a literature review, it was found that only a limited number of analytical methods—such as Ultraviolet spectroscopy, HPTLC, HPLC and fluorimetry — have been reported. Therefore, a novel, selective and sensitive Liquid approach was established for quantification of Bilastine in bulk and pharmaceutical dosage forms using Liquid Chromatography–Mass Spectrometry (LC-MS).

Analysis employed Agilent 1290 Infinity II LC system together with Agilent 6470 Triple Quadrupole Mass Spectrometer (or their equivalents). Chromatographic separation has been carried out on Poroshell Stable Bond (SB) C18 column (4.6×150mm, 2.7µm), using mobile phase composed of acetonitrile, water, methanol in a 40:30:30 ratio. Flow rate has been maintained at 0.5mL/min and detection has been carried out at 215nm with total run time of 6 minutes. The method has been confirmed as per International Council for Harmonisation (ICH) guidelines, encompassing parameters, namely specificity, accuracy, linearity, limit of detection (LOD), precision, limit of quantification (LOQ), robustness. The calibration curve showed linearity over a concentration range of 5–100µg/mL, with an LOD of 0.24µg/mL and an LOQ of 0.74µg/mL. All validation parameters met acceptable criteria, confirming the method's accuracy and precision.