Rapid Identification of Unknown Impurities in 3-Bromo-5-(trifluoromethyl)aniline by LC-SPE/NMR

Miroslava Harča,Iva Habinovec,Ernest Meštrović,Ivana Biljan,Predrag Novak

doi:10.5562/cca3042

Abstract

Identification of unknown pharmaceutical impurities is an essential part in the drug development process. In the present study, we developed and applied liquid chromatography (LC) – solid-phase extraction (SPE) / nuclear magnetic resonance (NMR) methodology with cryoprobe for identification and structural characterization of unknown impurities in 3- bromo-5-(trifluoromethyl)aniline. The three main impurities were separated and isolated by LC-SPE system. After multiple trapping, isolated impurities were eluted from the SPE cartridges with deuterated acetonitrile and one- and two- dimensional homo- and heteronuclear NMR spectra were recorded. The structures of the unknown impurities were determined by detailed inspection of NMR spectra and by mass spectrometric (MS) analysis. The results of the present preliminary study demonstrated that LC- SPE/NMR can be used for rapid impurity profiling of 3-bromo-5-(trifluoromethyl)aniline.

Highlights

A S impurities define the quality of a chemical entity, their unambiguous structure determination is compulsory part of the pharmaceutical development
The three main unknown impurities of 3-bromo-5-(trifluoromethyl)aniline were detected by DAD during the ultra high performance liquid chromatography (UHPLC) analysis eluting with respect to the main peak at relative retention times of
In order to elucidate the structures of the unknown impurities, compounds [1,2,3] were separated and isolated from the sample by liquid chromatography (LC)-solid-phase extraction (SPE) system

Summary

Introduction

A S impurities define the quality of a chemical entity, their unambiguous structure determination is compulsory part of the pharmaceutical development. We developed and applied LC-SPE/NMR methodology with cryoprobe for separation, isolation and structural characterization of the three main impurities in 3-bromo5-(trifluoromethyl)aniline sample. In order to increase the retention of the compounds on SPE cartridges a post-column water addition was performed with a flow rate of 3.0 mL min–1.

Results

Conclusion