title: Determination of thiols by capillary micellar electrokinetic chromatography with laser induced fluorescence detection using 1,3,5,7-tetramethyl-8-phenyl-(4-iodoacetamido) difluoroboradiaza-s-indacene as labeling reagent.
abstract: A sensitive and effective micellar electrokinetic capillary chromatography with laser-induced fluorescence detection approach was described for the determination of low molecular-mass thiols using 1,3,5,7-tetramethyl-8-phenyl-(4-iodoacetamido) difluoroboradiaza-s-indacene as the labeling reagent. After precolumn derivatization, baseline separation of six thiol compounds including cysteine, glutathione, N-acetylcysteine, homocysteine, 6-mercaptopurine, and penicillamine were achieved within 18 min. The optimal running buffer was composed of mixtures involving 25 mM sodium dodecyl sulfate, 25% (v/v) acetonitrile and 15 mM sodium phosphate buffer, pH 7.5. The detection limits (S/N = 3) were found as low as 40 pM under argon ion laser-induced fluorescence detector (λ(ex)/λ(em) = 488/520 nm), which were much better than the reported approaches. The accuracy and specificity of this assay for real samples were assured by a standard addition method. The proposed method has been applied to the analysis of thiols both in human plasma and plum flower samples with recoveries of 92.0-109.4%.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan, China.
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Guo
authors.forename: Hui-Xian
authors.initials: HX
authors.lastname: Zhang
authors.forename: Li-Na
authors.initials: LN
authors.lastname: Ma
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Hua-Shan
authors.initials: HS
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Guo
mesh_terms: D000081:Acetamides / Q000737:chemistry*; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000295:instrumentation / Q000379:methods*; D005453:Fluorescence; D006575:Heterocyclic Compounds, 3-Ring / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: C541287:1,3,5,7-tetramethyl-8-phenyl-(4-iodoacetamido)difluoroboradiaza-s-indacene; D000081:Acetamides; D006575:Heterocyclic Compounds, 3-Ring; D013438:Sulfhydryl Compounds