title: N-methyl-D-glucamine improves the laser-induced fluorescence capillary electrophoresis performance in the total plasma thiols measurement.
abstract: We describe an ultrarapid capillary electrophoresis with laser-induced fluorescence (CE-LIF) method for total plasma thiols measurement. Reduced thiols by 10% tri-n-butylphosphine (TBP) were derivatized in 10 min at room temperature with 5-iodoacetamidofluorescein (5-IAF) as fluorescent reagent. We show that CE-LIF allows a baseline separation of total plasma cysteinylglycine, homocysteine, cysteine, and glutathione in less than 5 min when N-methyl-D-glucamine in run buffer was added. CE was compared with high-performance liquid chromatography (HPLC) with fluorescence detection. The Bland-Altman test and Passing-Bablok regression demonstrates that the results obtained by CE-LIF and by HPLC are highly comparable. The simplified procedure of sample preparation, the short incubation and fast separation times, the high specificity, sensitivity and reproducibility, and the lower cost of analysis suggest that our proposed method can be considered valuable for the automation analysis in a routine laboratory.
authors.affiliation: Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Zinellu
authors.forename: Ciriaco
authors.initials: C
authors.lastname: Carru
authors.forename: Franca
authors.initials: F
authors.lastname: Galistu
authors.forename: Maria Franca
authors.initials: MF
authors.lastname: Usai
authors.forename: Giovanni Mario
authors.initials: GM
authors.lastname: Pes
authors.forename: Giovannella
authors.initials: G
authors.lastname: Baggio
authors.forename: Giorgio
authors.initials: G
authors.lastname: Federici
authors.forename: Luca
authors.initials: L
authors.lastname: Deiana
mesh_terms: D000328:Adult; D002021:Buffers; D003545:Cysteine / Q000097:blood / Q000302:isolation & purification; D004151:Dipeptides / Q000097:blood / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D005452:Fluoresceins; D005453:Fluorescence; D005978:Glutathione / Q000097:blood / Q000302:isolation & purification; D006710:Homocysteine / Q000097:blood / Q000302:isolation & purification; D006801:Humans; D007834:Lasers; D008536:Meglumine; D013438:Sulfhydryl Compounds / Q000097:blood* / Q000302:isolation & purification
chemical_list: D002021:Buffers; D004151:Dipeptides; D005452:Fluoresceins; D013438:Sulfhydryl Compounds; D006710:Homocysteine; C028505:cysteinylglycine; C015296:5-iodoacetamidofluorescein; D008536:Meglumine; D005978:Glutathione; D003545:Cysteine