titoli, abstracts, parole chiave >>>
Thiol redox status evaluation in red blood cells by capillary electrophoresis-laser induced fluorescence detection

Zinellu, Angelo and Sotgia, Salvatore and Usai, Maria Franca and Chessa, Roberto and Deiana, Luca and Carru, Ciriaco (2005) Thiol redox status evaluation in red blood cells by capillary electrophoresis-laser induced fluorescence detection. Electrophoresis, Vol. 26 (10), p. 1963-1968. Article.

Full text not available from this repository.

DOI: 10.1002/elps.200400042

Abstract

Thiols and in particular glutathione (GSH) play a central role in human metabolism, including the detoxification of xenobiotics, cell homeostasis, radioprotection, and antioxidant defence. Here, a new method is provided for the measurement of reduced and total forms of thiols in red blood cells. In order to minimize oxidation of reduced thiols, a water erythrocyte lysis (15 min at 4°C) was performed followed by a protein precipitation step with acetonitrile. The supernatant was rapidly derivatized with 5-iodoacetoamidefluorescein that trapped thiol groups, thus minimizing auto-oxidation. Derivatized samples were separated in a 57 cm x 75 µ m ID capillary by using 5 mmol/L sodium phosphate, 4 mmol/L boric acid as electrolyte solution with 75 mmol/L N-methyl-D-glucamine at pH 11.0. Under these conditions, cysteinylglycine (CysGly), cysteine (Cys), glutathione, and γ-glutamylcysteine (GluCys) were baseline-resolved in ̃ 4 min. Precision tests showed a good repeatability of our method both for migration times (coefficient of variation CV < 0.8%) and areas (CV < 3.3%). Furthermore, a good reproducibility of intrassay and interassay tests was obtained (CV migration times (coefficient of variation CV < 0.8%) and areas (CV < 3.3%). 5% and CV migration times (coefficient of variation CV < 0.8%) and areas (CV < 3.3%). 8%, respectively). The method was employed to investigate the effect of acidic precipitation on intracellular thiol concentration. Our data suggest that sample acidification causes a modification of the measured redox thiol status due to the development of a pro-oxidant environment; moreover, the thiol redox status of red blood cells was evaluated in 22 healthy volunteers.

Item Type:Article
ID Code:2436
Status:Published
Refereed:Yes
Uncontrolled Keywords:Capillary zone electrophoresis, 5-Iodoacetamidofluorescein, red blood cells, thiol oxidation, thiols
Subjects:Area 05 - Scienze biologiche > BIO/12 Biochimica clinica e biologia molecolare clinica
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
002 Altri enti e centri di ricerca del Nord Sardegna > Laboratorio analisi cliniche Meridiana Srl, Sassari
Publisher:Wiley
Deposited On:18 Aug 2009 10:07

I documenti depositati in UnissResearch sono protetti dalle leggi che regolano il diritto d'autore

Repository Staff Only: item control page