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Short-sweep capillary electrophoresis with a selective zinc fluorescence imaging reagent FluoZin-3 for determination of free and metalothionein-2a-bound Zn2+ ions

Citation

Nejdl, L and Moravanska, A and Smerkova, K and Mravec, F and Krizkova, S and Pomorski, A and Krezel, A and Macka, M and Adam, V and Vaculovicova, M, Short-sweep capillary electrophoresis with a selective zinc fluorescence imaging reagent FluoZin-3 for determination of free and metalothionein-2a-bound Zn2+ ions, Analytica Chimica Acta: International Journal Devoted to All Branches of Analytical Chemistry, 1017 pp. 41-47. ISSN 0003-2670 (2018) [Refereed Article]


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DOI: doi:10.1016/j.aca.2018.02.008

Abstract

A capillary electrophoretic (CE) method using a short-sweep approach and laser-induced fluorescence (LIF) detection (ShortSweepCE-LIF) was developed for determination of Zn2+ and Cd2+ as complexes with highly selective and sensitive fluorescent probe FluoZin-3. The ShortSweepCE-LIF method, established in this work, can be used for examining competitive Zn2+ and Cd2+ binding properties of metalloproteins or peptides. The parameters including background electrolyte composition, injection pressure and time as well as separation voltage were investigated. Under the optimized conditions, 80 mM HEPES, pH 7.4, with 1.5 μM FluoZin-3 was used as an electrolyte, hydrodynamic injection was performed at 50 mbar for 5 s, and separation voltage of 25 kV. Limits of detection for Zn2+ and Cd2+ were 4 and 125 nM, respectively. The developed method was demonstrated in a study of interactions between metalothionein-2a isoform and metal ions Zn2+, Co2+ and Cd2+. It was found that FluoZin-3 was able to extract a single Zn2+ ion, while added Co2+ (in surplus) extracted only 2.4 Zn2+ ions, and Cd2+ extracted all 7 Zn2+ ions present in the metalothionein molecule.

Item Details

Item Type:Refereed Article
Keywords:MEtal ions, zinc, cadmium, metalothionein, fluozin-3, capillary electrophoresis
Research Division:Chemical Sciences
Research Group:Analytical chemistry
Research Field:Separation science
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the chemical sciences
UTAS Author:Macka, M (Professor Mirek Macka)
ID Code:151526
Year Published:2018
Web of Science® Times Cited:5
Deposited By:Chemistry
Deposited On:2022-08-01
Last Modified:2022-08-01
Downloads:0

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