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J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Dec 15;941:62-8. doi: 10.1016/j.jchromb.2013.10.004. Epub 2013 Oct 11.

Determination of gibberellins in soybean using tertiary amine labeling and capillary electrophoresis coupled with electrochemiluminescence detection.

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1
College of Chemistry and Bioengineering, Guilin University of Technology, Guilin 541004, China.

Abstract

A novel sensitive method based on tertiary amine labeling for the analysis of gibberellins (GAs) by capillary electrophoresis (CE) coupled with electrochemiluminescence (ECL) detection was proposed. GA3 was tagged with 2-(2-aminoethyl)-1-methylpyrrolidine (AEMP) using N, N'-dicyclohexylcarbodiimide (DCC) and 3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine (HOOBt) as coupling agents in acetonitrile to produce GA3-AEMP-derivative. The GA3-AEMP-derivative was injected into CE by electrokinetic injection and detected by Ru(bpy)3(2+)-based ECL. The parameters affecting derivatization, detection and separation such as concentration of reactants, detection potential, pH and concentration of separation buffer, were investigated in detail. Under optimum conditions, the linear concentration range for GA3 was from 2.0×10(-7) to 1.28×10(-4)M with a correlation coefficient of 0.9997. The detection limit was 8×10(-8)M (S/N=3). The relative standard deviations of migration time, peak intensity and peak area for nine continuous injections of 2.0×10(-5)M GA3-AEMP-derivative were 1.0%, 2.1% and 4.2%, respectively. The developed approach was successfully applied to the determination of total GAs in the stem, leaf and seed of soybean (Glycine max [L.] Merr.) with recoveries in the range from 89.6% to 99.3%.

KEYWORDS:

Capillary electrophoresis; Electrochemiluminescence; Gibberellin A(3); Soybean; Tertiary amine labeling

PMID:
24184838
DOI:
10.1016/j.jchromb.2013.10.004
[Indexed for MEDLINE]
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