N ‐Methyl‐ D ‐glucamine improves the laser‐induced fluorescence capillary electrophoresis performance in the total plasma thiols measurement
2003; Wiley; Volume: 24; Issue: 16 Linguagem: Inglês
10.1002/elps.200305570
ISSN1522-2683
AutoresAngelo Zinellu, Ciriaco Carru, Franca Galistu, Maria Franca Usai, Giovanni Mario Pes, Giovannella Baggio, Giorgio Federici, Luca Deiana,
Tópico(s)Microfluidic and Capillary Electrophoresis Applications
ResumoAbstract 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.
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