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N-Methyl-D-glucamine improves the laser-induced fluorescence capillary electrophoresis performance in the total plasma thiols measurement

Zinellu, Angelo and Carru, Ciriaco and Galistu, Franca and Usai, Maria Franca and Pes, Giovanni Mario and Baggio, Giovannella and Federici, Giorgio and Deiana, Luca (2003) N-Methyl-D-glucamine improves the laser-induced fluorescence capillary electrophoresis performance in the total plasma thiols measurement. Electrophoresis, Vol. 24 (16), p. 2796-2804. eISSN 1522-2683. Article.

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DOI: 10.1002/elps.200305570

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.

Item Type:Article
ID Code:1189
Status:Published
Refereed:Yes
Uncontrolled Keywords:Capillary electrophoresis, cardiovascular risk, homocysteine, laser-induced fluorescence, N-Methyl-D-glucamine
Subjects:Area 05 - Scienze biologiche > BIO/12 Biochimica clinica e biologia molecolare clinica
Area 06 - Scienze mediche > MED/09 Medicina interna
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
001 Università di Sassari > 01 Dipartimenti > Neuroscienze, scienze materno infantili
Publisher:Wiley
eISSN:1522-2683
Deposited On:18 Aug 2009 10:04

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