Investigation of the oxidative transformation of roxarsone by electrochemistry coupled to hydrophilic interaction liquid chromatography/mass spectrometry

Frensemeier LM, Büter L, Vogel M, Karst U

Zusammenfassung

The oxidative transformation of the animal feed additive roxarsone (3-nitro-4-hydroxyphenylarsonic acid) was investigated using electrochemistry (EC) coupled to hydrophilic interaction liquid chromatography (HILIC) and mass spectrometry. Although roxarsone is claimed to be relatively stable in the organism, only little is known about its metabolism. By means of high-resolution electrospray (ESI)-MS analysis of electrochemically oxidized roxarsone, potential transformation products were identified including the toxic metabolite arsenate (As(V)). Thus, an oxidative carbon-arsenic bond cleavage was observed. For the first time, not only arsenic containing products were considered, but also non-arsenic species which were formed during As(V) generation and which could also be toxic metabolites of roxarsone. A hydrophilic interaction liquid chromatographic (HILIC) separation coupled to electrospray (ESI)-MS enabled polarity estimation and the identification of isomeric products. Additionally, hyphenation of HILIC to inductively coupled plasma (ICP)-MS allowed speciation analysis and quantification of arsenic containing products, revealing As(V) to be the main species formed under the applied oxidative conditions. The reactivity of the TPs towards biomolecules was investigated by means of adduct formation experiments. It has been found that two electrochemically generated species, one quinone and one quinone imine, showed reactivity towards free thiol groups of the tripeptide glutathione and the proteins beta-lactoglobulin A and human serum albumin, which were used as model compounds for adduct formation.