A High-Performance Liquid Chromatography Method for Determination of Genotoxic Impurity Hydroxylamine in Drug Substances

摘要

Hydroxylamine (NH2OH) is widely used in pharmaceutical intermediates and final drug substances synthesis. Since hydroxylamine is a well-known genotoxic impurity compound that needs to be controlled down to ppm level in pharmaceutical compounds. It is very difficult to detect using conventional analytical techniques due to its physical-chemical properties like the lack of chromophore, low molecular weight, the absence of carbon atom and high polarity. In addition to that, the analysis of the pharmaceutical samples encounters considerable obstruction from matrix components that greatly overshadow the response of hydroxylamine. This report describes a simple, selective and sensitive high-performance liquid chromatography (HPLC)-UV derivatization method for the determination of hydroxylamine in drug substances. The HPLC method was detected up to 0.01 ppm of hydroxylamine with S/N > 3.0 and quantified up to 0.03 ppm of hydroxylamine with S/N ratio > 10.0. This validated method can be applied as a generic method to detect the hydroxylamine for pharmaceutical process control and drug substance release.