Development and Validation of a Liquid Chromatography-Tandem Mass Spectrometry Method for the Simultaneous Determination of N-NitrosoHydrochlorothiazide and N-Nitroso-Bisoprolol in Bisoprolol Hydrochlorothiazide Fixed-Dose Combination Tablets
Year: 2026
Type: article
Abstract: Nitrosamine impurities in pharmaceutical products pose serious carcinogenic risks requiring stringent analytical control. This work describes a comprehensive validated liquid chromatography-tandem mass spectrometry method for simultaneous quantification of N-nitroso-hydrochlorothiazide and N-nitroso-bisoprolol in fixed-dose combination tablets. Chromatographic separation utilized a Kinetex Biphenyl column with gradient elution combining 5 mM ammonium formate with 0.1% formic acid in water and 0.1% formic acid in methanol. Mass spectrometric detection employed multiple reaction monitoring with dual polarity electrospray ionization (positive mode for N-nitroso-bisoprolol m/z 377.10 → 275.15; negative mode for N-nitroso-hydrochlorothiazide m/z 325.00 → 293.95). Validation following ICH Q2(R2) guidelines demonstrated excellent performance: limits of quantification at 798 ppm and 15 ppm respectively (approximately 10% of specification limits), linearity with correlation coefficients exceeding 0.999, mean recoveries of 100.9% ± 2.2% and 100.8% ± 2.5%, precision with relative standard deviation values below 3.7%, and robust performance under deliberately varied conditions. This validated method provides a reliable analytical tool for pharmaceutical quality control and regulatory compliance with European Medicines Agency and Food and Drug Administration standards for nitrosamine impurity monitoring.
Authors: Kiran Kumar Kurella, Anima S. Dadhich
Institution: GITAM University
Language: English
Subfield: Analytical Chemistry
Field: Chemistry
Domain: Physical Sciences
Open Access status: bronze
OpenAlex ID: W7160390473