Highly sensitive and selective detection of biothiols using graphene oxide-based "molecular beacon"-like fluorescent probe.
A fluorometric method for quantity analysis of biothiols was developed using a graphene oxide (GO)-based "molecular beacon"-like probe, which consisted of FITC labeled thymine (T)-rich single-stranded DNA (ssDNA), GO and Hg(2+) ions. The labeled ssDNA containing T-T mismatches would self-hybridize to duplex in the presence of Hg(2+), which can avoid its adsorption on GO and the fluorescence of this GO-based probe was recovered. The fluorescence of the probe quenched after the addition of biothiols such as glutathione (GSH) and cysteine (Cys) owing to thiol groups can selectively competitive ligation of Hg(2+) ions with T-T mismatches. In the present work, the GO-based probe was used for the determination of GSH and Cys. Under the optimal conditions, a linear correlation was established between fluorescence intensity ratio I(0)/I and the concentration of GSH in the range of 2.0×10(-9)-5.0×10(-7) mol L(-1) with a detection limit of 1.0×10(-9) mol L(-1). The linear range for Cys is from 5.0×10(-9) to 4.5×10(-7) mol L(-1) with a detection limit of 2.0×10(-9) mol L(-1). The proposed method was applied to the determination of GSH in human serum and cell extract samples with satisfactory results.
Department of Analytical Chemistry, College of Chemistry, Jilin University, Changchun 130012, China. gaoyuan_jlu@yahoo.com.cn
Yuan
Y
Gao
Yan
Y
Li
Xin
X
Zou
Hui
H
Huang
Xingguang
X
Su
D020137:Base Pair Mismatch; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D003545:Cysteine / Q000032:analysis / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000235:genetics; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D005470:Fluorometry; D005978:Glutathione / Q000032:analysis / Q000737:chemistry; D006108:Graphite / Q000737:chemistry*; D056945:Hep G2 Cells; D006801:Humans; D008628:Mercury / Q000737:chemistry; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D010087:Oxides / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000032:analysis* / Q000737:chemistry
D015342:DNA Probes; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D010087:Oxides; D013438:Sulfhydryl Compounds; D006108:Graphite; D008628:Mercury; D005978:Glutathione; D016650:Fluorescein-5-isothiocyanate; D003545:Cysteine