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High-throughput capillary electrophoresis method for plasma cysteinylglycine measurement: evidences for a clinical application

Zinellu, Angelo and Pinna, Antonio <1963- > and Zinellu, Elisabetta and Sotgia, Salvatore and Deiana, Luca and Carru, Ciriaco (2008) High-throughput capillary electrophoresis method for plasma cysteinylglycine measurement: evidences for a clinical application. Amino Acids, Vol. 34 (1), p. 69-74. eISSN 1438-2199. Article.

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DOI: 10.1007/s00726-007-0590-4

Abstract

Increased levels in plasma homocysteine and cysteine, and more recently, decreased levels in cysteinylglycine have been indicated as a risk factor for vascular diseases. Most assays focused their attention only on homocysteine determination and when also other thiols were measured, analytical times drastically increased. By modifying our previous method for thiols detection, we set up a rapid capillary electrophoresis method for the selective quantification of plasma cysteinylglycine, cutting the analysis time of about 50%. Samples were treated with tri-n-butylphosphine as reducing agent, proteins were precipitated with trichloroacetic acid and released thiols were successively derivatized by the selective thiol laser-induced fluorescence-labeling agent 5-iodoacetamidofluorescein and separated by capillary electrophoresis. A baseline separation between peaks was obtained in about 2 min using 3 mmol/L sodium phosphate/2.5 mmol/L boric acid as electrolyte solution with 75 mmol/L N-methyl-D-glucamine at pH 11.25 in a 47 cm long capillary with a cartridge temperature of 45 °C. The method application was checked by measuring plasma Cys-Gly levels in a group of patients affected by retinal vein occlusion (RVO), an important cause of visual loss in the elderly. The low levels of Cys-Gly found in the RVO patients suggest that these small thiols may have importance in the disease development.

Item Type:Article
ID Code:3704
Status:Published
Refereed:Yes
Uncontrolled Keywords:Cysteinylglycine, cysteine, homocysteine, capillary electrophoresis, retinal vein occlusion
Subjects:Area 06 - Scienze mediche > MED/30 Malattie apparato visivo
Area 05 - Scienze biologiche > BIO/12 Biochimica clinica e biologia molecolare clinica
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
001 Università di Sassari > 03 Istituti > Oftalmologia, Otorinolaringoiatria e Urologia
002 Altri enti e centri di ricerca del Nord Sardegna > National laboratory of the national institute of biostructures and biosystems, Osilo
Publisher:Springer Wien
eISSN:1438-2199
Deposited On:09 Apr 2010 12:46

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