N-methyl-D-glucamine improves the laser-induced fluorescence capillary electrophoresis performance in the total plasma thiols measurement.
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.
Zinellu
Angelo
A
Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy.
Carru
Ciriaco
C
Galistu
Franca
F
Usai
Maria Franca
MF
Pes
Giovanni Mario
GM
Baggio
Giovannella
G
Federici
Giorgio
G
Deiana
Luca
L
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
D002021:Buffers; D004151:Dipeptides; D005452:Fluoresceins; D013438:Sulfhydryl Compounds; D006710:Homocysteine; C028505:cysteinylglycine; C015296:5-iodoacetamidofluorescein; D008536:Meglumine; D005978:Glutathione; D003545:Cysteine