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Albumin-bound low molecular weight thiols analysis in plasma and carotid plaques by CE

Zinellu, Angelo and Lepedda, Antonio Junior and Sotgia, Salvatore and Zinellu, Elisabetta and Marongiu, Giommaria and Usai, Maria Franca and Gaspa, Leonardo and De Muro, Pierina and Formato, Marilena and Deiana, Luca and Carru, Ciriaco (2010) Albumin-bound low molecular weight thiols analysis in plasma and carotid plaques by CE. Journal of Separation Science, Vol. 33 (1), p. 126-131. eISSN 1615-9314. Article.

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DOI: 10.1002/jssc.200900582

Abstract

We describe a new method for the quantification of low molecular weight thiols, as homocysteine, cysteine, cysteinylglycine, glutamylcysteine and glutathione bound to human plasma albumin. After albumin isolation and purification by SDS-PAGE, thiols were freed from protein with tri-n-butylphosphine and successively derivatized with 5-iodoacetamidofluorescein. Samples were then injected and quantified in about 18 min by CE with laser induced fluorescence detection. Precision tests indicate a good repeatability of the method both for migration times (RSD<0.63%) and areas (RSD<2.98%). The method allows to measure all five low molecular weight thiols released from just 3 μg of albumin thus improving the other described methods in which only three or four thiols were detected. Due to the elevated sensitivity (LOD of 0.3 pM for all thiols), also low molecular weight thiols bound to albumin filtered in tissues could be quantified.

Item Type:Article
ID Code:5302
Status:Published
Refereed:Yes
Uncontrolled Keywords:Albumin, CE, cysteine, homocysteine, SDS-PAGE
Subjects:Area 06 - Scienze mediche > MED/22 Chirurgia vascolare
Area 05 - Scienze biologiche > BIO/10 Biochimica
Area 05 - Scienze biologiche > BIO/12 Biochimica clinica e biologia molecolare clinica
Divisions:001 Università di Sassari > 02 Centri > Centro di eccellenza interdisc. sviluppo ricerca biotecnologica e studio biodiversità della Sardegna e dell'area mediterranea
001 Università di Sassari > 01 Dipartimenti > Scienze fisiologiche, biochimiche e cellulari
001 Università di Sassari > 03 Istituti > Clinica chirurgica generale e terapia chirurgica
001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
002 Altri enti e centri di ricerca del Nord Sardegna > National laboratory of the national institute of biostructures and biosystems, Osilo
Publisher:Wiley
eISSN:1615-9314
Copyright Holders:© 2009 WILEY-VCH Verlag
Deposited On:08 Feb 2011 11:13

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