Microwave photochemical reactor for the online oxidative decomposition of p-hydroxymercurybenzoate (pHMB)-tagged proteins and their determination by cold vapour generation atomic fluorescence detection

Year: 2013

Authors: Campanella B., Gonzalez Rivera J., Ferrari C., Biagi S., Onor M., D’Ulivo A., Bramanti E.

Autors Affiliation: National Research Council of Italy, C.N.R., Istituto di Chimica dei Composti Organo Metallici − ICCOM − UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa, Italy;
Chemical Engineering Department, University of Guanajuato, Noria Alta s/n 36050 Guanajuato, Gto., Mexico;
National Research Council of Italy, C.N.R., Istituto Nazionale di Ottica, INO − UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa, Italy

Abstract: A novel method is presented for the characterization and determination of thiolic proteins. After the labeling with p-hydroxymercurybenzoate, the pHMB-labeled proteins underwent on-line oxidation with a novel microwave (MW)/UV photochemical reactor, followed by cold vapor generation-atomic fluorescence spectrometry (CVG-AFS) detection. The MW/UV process led to the conversion of pHMB to Hg(II)with a yield of 89.0 ± 0.5% without using chemical oxidizing reagents and avoiding the use of toxic carcinogenic compounds. Hg(II) was reduced to Hg0 in a knotted reaction coil with NaBH4 solution, stripped from the solution by an argon flow and detected. The chromatographic method for labeled thiolic peptides was linear in the 0.2−100 μmol L−1 range, with a LOD as mercury of 57 nmol L−1. This system has proven to be a useful interface for liquid chromatography coupled with CVG-AFS in the determination and characterization of thiolic proteins. This method has been applied to the determination of thiolic peptides after tryptic digestion of serum albumins from different species (human, bovine, rat, horse, and sheep).


Volume: 85 (24)      Pages from: 12152  to: 12157

KeyWords: Microwave chemistry, photochemical reaction, proteins, Atomic Fluorescence
DOI: 10.1021/ac403389z

Citations: 6
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