Evaluation of Various Potential Genotoxic Nitrosamine Impurities by Using Validated Ultra - Sensitive LC/MS/MS Analytical Method in Anti-Hypertensive Drug Product.

Abstract

Currently, genotoxic impurities are a prominent concern among other impurities, and regulatory authorities are focused more on identifying, quantifying, and controlling these undesired chemicals in drug substances and drug products. ICH M7 provides an overview of the class of genotoxic impurities based on the potency category. To meet the regulatory requirement for the concerned impurity, a sensitive analytical method capable of quantifying these impurities at a lower level with accuracy and precision is required. This article focuses on the development of an analytical method for NDMA (N-Nitroso dimethylamine), NDEA (N-Nitroso diethylamine), NEIPA (N-Nitrosoethylisopropylamine), NDIPA (N-Nitroso diisopropylamine), NDPA (N-Nitroso dipropylamine) and NDBA (N-Nitroso dibutylamine)  nitrosamine impurities in a single chromatography method by LC-MS/MS positive mode of atmospheric pressure chemical ionization (APCI), employing multiple reaction monitoring (MRM) with a limit of quantification (LOQ) of 29 parts per billions (ppb) for NDEA, NEIPA, NDIPA, NDPA & NDBA against the acceptance limit of 331 ppb and for NDMA, the LOQ is 107.77 ppb against the acceptance limit of 1200 ppb. The chromatographic separation was accomplished using an Agilent Poroshell 120EC- C18 (150 mm × 4.6 mm), 2.7 mm utilizing a gradient mode elution program including mobile phase A (0.1% formic acid in water) and mobile phase B (4mM ammonium acetate solution in methanol). The method was challenged for accuracy, precision, and linearity in accordance with ICH guidelines to ensure its suitability for the intended usage.