Title: Thiol redox status evaluation in red blood cells by capillary electrophoresis-laser induced fluorescence detection
Abstract: ELECTROPHORESISVolume 26, Issue 10 p. 1963-1968 Research Article Thiol redox status evaluation in red blood cells by capillary electrophoresis-laser induced fluorescence detection Angelo Zinellu, Corresponding Author Angelo Zinellu [email protected] Chair of Clinical Biochemistry, University of Sassari Angelo Zinellu, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120=== Ciriaco Carru, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120===Search for more papers by this authorSalvatore Sotgia, Salvatore Sotgia Chair of Clinical Biochemistry, University of SassariSearch for more papers by this authorMaria Franca Usai, Maria Franca Usai Laboratory for Clinical Analysis, Meridiana srl, Sassari, ItalySearch for more papers by this authorRoberto Chessa, Roberto Chessa Chair of Clinical Biochemistry, University of SassariSearch for more papers by this authorLuca Deiana, Luca Deiana Chair of Clinical Biochemistry, University of SassariSearch for more papers by this authorCiriaco Carru, Corresponding Author Ciriaco Carru [email protected] Chair of Clinical Biochemistry, University of Sassari Angelo Zinellu, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120=== Ciriaco Carru, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120===Search for more papers by this author Angelo Zinellu, Corresponding Author Angelo Zinellu [email protected] Chair of Clinical Biochemistry, University of Sassari Angelo Zinellu, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120=== Ciriaco Carru, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120===Search for more papers by this authorSalvatore Sotgia, Salvatore Sotgia Chair of Clinical Biochemistry, University of SassariSearch for more papers by this authorMaria Franca Usai, Maria Franca Usai Laboratory for Clinical Analysis, Meridiana srl, Sassari, ItalySearch for more papers by this authorRoberto Chessa, Roberto Chessa Chair of Clinical Biochemistry, University of SassariSearch for more papers by this authorLuca Deiana, Luca Deiana Chair of Clinical Biochemistry, University of SassariSearch for more papers by this authorCiriaco Carru, Corresponding Author Ciriaco Carru [email protected] Chair of Clinical Biochemistry, University of Sassari Angelo Zinellu, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120=== Ciriaco Carru, Chair of Clinical Biochemistry, University of Sassari, Viale San Pietro 43/B, I-07100 Sassari, Italy Fax: +39-079228120===Search for more papers by this author First published: 04 May 2005 https://doi.org/10.1002/elps.200400042Citations: 31AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat 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 × 75 μm 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 < 5% and CV < 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. Citing Literature Volume26, Issue10No. 10 May 2005Pages 1963-1968 RelatedInformation
Publication Year: 2005
Publication Date: 2005-05-01
Language: en
Type: article
Indexed In: ['crossref', 'pubmed']
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Cited By Count: 40
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