Capillary electrophoresis fingerprinting of 8-aminopyrene-1,3,6-trisulfonate derivatized nitrocellulose after partial acid depolymerization.
Elodie Alinat, Nathalie Delaunay, Cédric Przybylski, Régis Daniel, Xavier Archer, Pierre Gareil
文献索引:J. Chromatogr. A. 1387 , 134-43, (2015)
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摘要
Fine characterization of nitrocellulose (NC) remains a challenge, especially in forensic analysis, and a strategy consisting in obtaining representative fingerprints by a separation technique, as for proteins, is of prime interest. In this work, we first established that NCs (especially of high molar mass) cannot be representatively derivatized by 8-aminopyrene-1,3,6-trisulfonic acid (APTS), because of their poor solubility in the medium required for APTS derivatization. Therefore, a partial acid depolymerization step was considered, prior to derivatization by APTS, in an attempt to generate a mixture of oligosaccharides retaining information on the initial NC sample and/or on the cellulose used to prepare it. Acid depolymerization conditions (time and acid concentration) as well as APTS derivatization conditions (time, temperature, APTS/NC and reducing agent/APTS molar ratios) were investigated for lowly-nitrated NCs. The best compromise between depolymerization yields, speed, and pertinency of the resulting oligosaccharidic mixture was obtained using fuming hydrochloric acid (37%, w/w) at 50 °C for 30 min. The most effective procedure for APTS derivatization of oligosaccharides obtained after partial acid depolymerization of NC was achieved at 70 °C for 2h. The resulting APTS-derivatized oligosaccharides were then separated by capillary electrophoresis (CE) using a background electrolyte composed of 60mM 6-aminocaproic acid, pH 4.5 (adjusted with acetic acid)+0.02% hydroxypropyl methyl cellulose. Finally, for the first time, they were identified using APTS-derivatized cellodextrin standards and by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS).Copyright © 2015 Elsevier B.V. All rights reserved.
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