title: Stability-indicating HPLC-UV for the determination of 4-bromomethyl-3-nitrobenzoic acid, a bioactive nitrocompound.
abstract: The nitroaromatic compound 4-bromomethyl-3-nitrobenzoic acid (ANB) is a promising antitumoral agent whose activity has recently been investigated. Forced degradation studies were conducted on ANB with a high-performance liquid chromatography-ultraviolet assay to establish its stability and selectivity. ANB was subjected to degradation studies under hydrolytic (acid and alkaline), oxidative and light exposition conditions. The compound showed greater lability only in acid and alkaline conditions by forming a major degradation product. The chromatographic separation of ANB and its degradation product was achieved on an octadecylsilane column using a mobile phase of methanol-water (80:20, v/v), pH 4.0 adjusted with formic acid, flow rate of 0.7 mL/min, ultraviolet diode array detection at λ 271 nm, injection volume of 20 µL and temperature of 30°C. The method was validated according to International Conference on Harmonization guidelines with respect to precision (average relative standard deviation 0.67%), accuracy (average 99.97%), linearity (y = 118730x + 12912; coefficient of determination > 0.999), specificity and robustness (p > 0.05). The degradation product formed as a result from the hydrolysis of nitrobenzyl bromide, corresponded to its benzyl alcohol, 4-hydroxymethyl-3-nitrobenzoic acid (ANOH) and was characterized by co-elution with a synthetic ANOH working standard.
authors.affiliation: Department of Pharmaceutical Products, Faculty of Pharmacy, Federal University of Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil betanialf@hotmail.com.
authors.forename: Maria Betânia
authors.initials: MB
authors.lastname: de Freitas
authors.affiliation: Department of Pharmaceutical Products, Faculty of Pharmacy, Federal University of Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil.
authors.forename: Eduardo Burgarelli
authors.initials: EB
authors.lastname: Lages
authors.affiliation: Department of Pharmaceutical Products, Faculty of Pharmacy, Federal University of Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil.
authors.forename: Isadora Marques Brum
authors.initials: IM
authors.lastname: Gonçalves
authors.affiliation: Department of Pharmaceutical Products, Faculty of Pharmacy, Federal University of Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil.
authors.forename: Renata Barbosa
authors.initials: RB
authors.lastname: de Oliveira
authors.affiliation: Department of Pharmaceutical Products, Faculty of Pharmacy, Federal University of Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil.
authors.forename: Cristina Duarte
authors.initials: CD
authors.lastname: Vianna-Soares
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D009579:Nitrobenzoates / Q000032:analysis* / Q000737:chemistry*; D015203:Reproducibility of Results
chemical_list: D009579:Nitrobenzoates; C075153:4-bromomethyl-3-nitrobenzoic acid