RESUMO
In this paper, titanium dioxide nanoparticles (TiO(2) NPs) were employed to construct a functional film on PDMS microfluidic channel surface, which was formed by sequentially immobilizing poly(diallyldimethylammonium chloride) and TiO(2) NPs on PDMS surface by layer-by-layer assembly technique. The modified PDMS microchip exhibited a decreased and stable EOF, which was favorable for the separation of biomolecules with similar migration times. Arginine, phenylalanine, serine and threonine were used as model analytes to evaluate the performance of the modified microchip. The four amino acids were efficiently separated within 100 s in a 3.7 cm long separation channel and successfully detected on the carbon fiber electrode in conjunction with in-channel indirect amperometry. Resolutions and theoretical plate numbers of the analytes were considerably enhanced in the presence of TiO(2) NPs. The modified microchip demonstrated excellent stability and reproducibility with improved RSDs of migration times and peak currents for run-to-run, day-to-day and chip-to-chip analyses, respectively. Variables influencing the separation efficiency and amperometric response, including injection and separation voltage, the working electrode position and buffer concentration, were optimized in detail.
Assuntos
Aminoácidos/análise , Dimetilpolisiloxanos/química , Eletroforese em Microchip/instrumentação , Nanopartículas/química , Titânio/química , Eletroforese em Microchip/métodos , Desenho de Equipamento , Limite de Detecção , Reprodutibilidade dos Testes , Propriedades de SuperfícieRESUMO
For the first time, a simple and 'green' approach based on one-step strategy was designed and developed for the modification of a fused-silica capillary with polynorepinephrine (PNE) to separate amino acid enantiomers using capillary electrochromatography coupled with electrogenerated chemiluminescence detection (CEC-ECL). Norepinephrine (NE) was filled into capillary to generate PNE coating on the surface of capillary as permanent coating via the oxidation of NE by oxygen dissolvable in the solution. The formation of the PNE coating was characterized by scanning electron microscopy, UV-vis spectra and contact angle measurements. Compared with the native capillary, the modified capillary had much better wettability, less nonspecific adsorption toward amino acids, and the enantiomers of histidine, phenylalanine, and valine samples received baseline separation with the resolution factors of 1.6, 1.8 and 1.6, respectively, utilizing a separation length of 40 cm of the capillary coupled with ECL detection on the PNE-coated capillary.
Assuntos
Eletrocromatografia Capilar/instrumentação , Medições Luminescentes/métodos , Norepinefrina/química , Polímeros/química , Aminoácidos/análise , Aminoácidos/química , Aminoácidos/isolamento & purificação , Eletrocromatografia Capilar/métodos , Limite de Detecção , Norepinefrina/análogos & derivados , Polimerização , Polímeros/síntese química , Reprodutibilidade dos Testes , EstereoisomerismoRESUMO
A novel, simple, and economical method for the preparation of chiral stationary phases for chip-based enantioselective open tubular capillary electrochromatography (OT-CEC) using polydopamine (PDA) coating as an adhesive layer was reported for the first time. After the poly(dimethylsiloxane) (PDMS) microfluidic chip was filled with dopamine (DA) solution, PDA film was gradually formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of DA by the oxygen dissolved in the solution. Due to possessing plentiful catechol and amine functional groups, PDA coating can serve as a versatile multifunctional platform for further secondary reactions, leading to tailoring of the coatings for protein bioconjugation by the thiols and amines via Michael addition or Schiff base reactions. Bovine serum albumin (BSA), acting as a target protein, was then stably and homogeneously immobilized in the PDA-coated PDMS microchannel to fabricate a novel protein stationary phase. Compared with the native PDMS microchannels, the modified surfaces exhibited much better wettability, more stable and enhanced electroosmotic mobility, and less nonspecific adsorption. The water contact angle and electroosmotic flow of PDA/BSA-coated PDMS substrate were measured to be 44° and 2.83×10(-4)cm(2)V(-1)s(-1), compared to those of 112° and 2.10×10(-4)cm(2)V(-1)s(-1) from the untreated one, respectively. Under a mild condition, d- and l-tryptophan were efficiently separated with a resolution of 1.68 within 130s utilizing a separation length of 37mm coupled with in-column amperometric detection on the PDA/BSA-coated PDMS microchips. This present versatile platform, facile conjugation of biomolecules onto microchip surfaces via mussel adhesive protein inspired coatings, may offer new processing strategies to prepare a biomimetic surface design on microfluidic chips, which is promising in high-throughput and complex biological analysis.