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1.
Methods Mol Biol ; 1274: 161-7, 2015.
Article in English | MEDLINE | ID: mdl-25673491

ABSTRACT

Electrochromatography (EC) on a porous monolithic stationary phase prepared within the channels of a microsystem is an attractive alternative for on-chip separation. It combines the separation mechanisms of electrophoresis and liquid chromatography. Moreover, the porous polymer monolithic materials have become popular as stationary phase due to the ease and rapidity of fabrication via free radical photopolymerization. Here, we describe a hexyl acrylate (HA)-based porous monolith which is simultaneously in situ synthesized and anchored to the inner walls of the channel of a cyclic olefin copolymer (COC) device in only 2 min. The baseline separation of a mixture of neurotransmitters including six amino acids and two catecholamines is realized.


Subject(s)
Acrylates/chemistry , Capillary Electrochromatography/methods , Cycloparaffins/chemistry , Polymers/chemistry
2.
J Chromatogr A ; 1376: 159-66, 2015 Jan 09.
Article in English | MEDLINE | ID: mdl-25537173

ABSTRACT

This work aims at studying the potentialities of an on-line capillary electrophoresis (CE)-based digestion methodology for evaluating polymer-drug conjugates degradability in the presence of free trypsin (in-solution digestion). A sandwich plugs injection scheme with transverse diffusion of laminar profile (TDLFP) mode was used to achieve on-line digestions. Electrophoretic separation conditions were established using poly-l-Lysine (PLL) as reference substrate. Comparison with off-line digestion was carried out to demonstrate the feasibility of the proposed methodology. The applicability of the on-line CE-based digestion methodology was evaluated for two PLL-drug conjugates and for the four first generations of dendrigraft of lysine (DGL). Different electrophoretic profiles presenting the formation of di, tri, and tetralysine were observed for PLL-drug and DGL. These findings are in good agreement with the nature of the linker used to link the drug to PLL structure and the predicted degradability of DGL. The present on-line methodology applicability was also successfully proven for protein conjugates hydrolysis. In summary, the described methodology provides a powerful tool for the rapid study of biodegradable polymers.


Subject(s)
Pharmaceutical Preparations/chemistry , Polylysine/chemistry , Animals , Cattle , Dendrimers , Diffusion , Electrophoresis, Capillary , Hydrolysis , Polymers , Serum Albumin, Bovine/chemistry , Taurine/chemistry , Trypsin/chemistry
3.
J Chromatogr A ; 1217(51): 8001-8, 2010 Dec 17.
Article in English | MEDLINE | ID: mdl-20800231

ABSTRACT

In order to develop a portable and disposable instrument for on-site analysis of neutral compounds, a lauryl methacrylate monolith has been synthesized into a cyclic olefin copolymer microdevice for reversed-phase electrochromatography purposes. This monolith was tested in capillary to evaluate electrochromatographic performances in terms of electroosmotic flow (EOF) mobility, retention and efficiency prior to its transfer into the microfluidic device. The produced monolithic bed exhibited a good run-to-run repeatability, column-to-column reproducibility and batch-to-batch reproducibility, with relative standard deviation (RSD) values below 9% for EOF mobility, retention factors and heights of theoretical plate. The electrochromatographic performances of the monolith were optimized by reducing irradiation time. Photopolymerization time of 10 min was found to be the best process in order to obtain a robust, retentive and efficient system. The on-chip performances of this monolith were evaluated in detail for the reversed-phase electrochromatographic separation of polycyclic aromatic hydrocarbons, with plate heights reaching down to 15 µm when working at optimal velocity. Aiming at the maximum simplification of instrumental fabrication and operation, a direct injection from a 2 µL droplet was compared with more conventional dynamic injection process.


Subject(s)
Capillary Electrochromatography/methods , Cycloparaffins/analysis , Polymers/analysis , Capillary Electrochromatography/instrumentation
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