Journal of chromatography. A 2012-01-13

Selective extraction of thiol-containing peptides in seawater using Tween 20-capped gold nanoparticles followed by capillary electrophoresis with laser-induced fluorescence.

Chien-Chih Shen, Wei-Lung Tseng, Ming-Mu Hsieh

Index: J. Chromatogr. A. 1220 , 162-8, (2012)

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Abstract

This study combines Tween 20-capped gold nanoparticles (Tween 20-AuNPs) with capillary electrophoresis (CE) for ultrasensitive detection of thiol-containing peptides, including glutathione (GSH), γ-glutamylcysteine (γ-GCS), and phytochelatin analogs. By forming AuS bonds, Tween 20-AuNPs can selectively extract and enrich these thiols from a complicated matrix. A Tween 20 capping layer not only suppresses nonspecific adsorption, but also enables NPs to disperse in a highly salinity solution. Dithiothreitol removes thiol-containing peptides from the NP surface through ligand exchange. The released peptides are selectively derivatized with o-phthaldialdehyde (OPA) to form tricyclic isoindole derivatives. Extraction efficiency of five thiol-containing peptides with Tween 20-AuNPs was highly reliable in the Tween 20-AuNP concentration, time of extraction and desorption thiols, and sample volume. After injecting a large sample volume, the OPA-derivatized peptides migrate against the electroosmotic flow (EOF) and enter the polyethylene oxide (PEO) zone. The sensitivity of these peptides was improved by stacking them at the boundary between the sample and PEO zones. As a result, limits of detection (LODs) for five peptides were down to 0.1-6 pM. Not only is the proposed method probably the first CE example for detecting dissolved thiols in seawater; it also has the lowest LODs for GSH, γ-GCS, and phytochelatins compared to other reported methods.Copyright © 2011 Elsevier B.V. All rights reserved.