Enantiomeric separation of tryptophan by open-tubular microchip capillary electrophoresis using polydopamine/gold nanoparticles conjugated DNA as stationary phase

Abstract

We reported the use of polydopamine/Au nanoparticles/DNA (PDA/Au NPs/DNA) as a stationary phase for the separation of tryptophan enantiomers by microchip capillary electrophoresis (MCE). The stationary phase was formed by sequentially immobilizing PDA/Au NPs and DNA to the glass microfluidic channel surface using layer-by-layer assembly. The formation of PDA/Au NPs as a functional film involved the use of HAuCl₄ as an oxidant to trigger DA self-polymerization. DA was then used as a reductant to reduce HAuCl₄ to Au NPs. All these Au NPs were simultaneously deposited on the microchip surface by the adhesive PDA, and the thiolated DNA was strongly adsorbed onto the glass microfluidic channel surface through covalent gold–sulfur bonding. The functional film of the PDA/Au NPs/DNA was characterized by scanning electron microscopy, UV-vis spectra and ATR-FT-IR. The influence of separation voltage and the concentration and pH value of the running buffer were found to significantly influence separation. Successful baseline separation of tryptophan enantiomers was achieved within 65 s utilizing an effective separation length of 33 mm, and coupling with laser-induced fluorescence detection resulted in a resolution factor of 2.95. All these evaluation parameters, including stability and reproducibility, were considered satisfactory. The method outlined here, using the PDA/Au NPs/DNA-coated microfluidic channel as a versatile platform, may offer a processing strategy to prepare a functional surface on microfluidic chips.