Simultaneous determination of cystine and other free aminothiols in blood plasma using capillary electrophoresis with pH-mediated stacking.

Autor: Ivanov AV; Department of Molecular and Cell Pathophysiology, Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Popov MA; Moscow Regional Research and Clinical Institute n.a. M.F. Vladimirskiy, Moscow, Russia., Aleksandrin V; Department of Molecular and Cell Pathophysiology, Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Pudova PA; Department of Molecular and Cell Pathophysiology, Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Galdobina MP; Department of Molecular and Cell Pathophysiology, Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Metelkin AA; Department of Molecular and Cell Pathophysiology, Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Kruglova MP; Federal State Autonomous Educational Institution of Higher Education I.M. Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University), Moscow, Russia., Maslennikov RA; Moscow Regional Research and Clinical Institute n.a. M.F. Vladimirskiy, Moscow, Russia., Silina EV; Federal State Autonomous Educational Institution of Higher Education I.M. Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University), Moscow, Russia., Stupin VA; Pirogov Russian National Research Medical University (RNRMU), Moscow, Russia., Kubatiev AA; Department of Molecular and Cell Pathophysiology, Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia.
Jazyk: angličtina
Zdroj: Electrophoresis [Electrophoresis] 2024 Mar; Vol. 45 (5-6), pp. 411-419. Date of Electronic Publication: 2023 Dec 12.
DOI: 10.1002/elps.202300196
Abstrakt: We developed a method of sensitive capillary electrophoresis using UV detection for the determination of certain free aminothiols (reduced cysteinylglycine (rCysGly), cysteine (rCys), glutathione (rGln), and cystine (CysS) in human blood plasma. The reduced thiols were derivatized with N-ethylmaleimide. The plasma was purified from proteins via ultrafiltration. Electrophoretic separation was performed using 115 mM Na phosphate with 7.5% (v/v) polyethylene glycol 600, pH 2.3. The in-capillary concentration of the analytes was achieved with a pH gradient created via the preinjection of triethanolamine and postinjection of phosphoric acid. The separation was carried out using a silica capillary (50 µm i.d.; total/effective separation length 42/35 cm) at a 25 kV voltage. The total analysis/regeneration time was 18 min. The quantification limits varied from 1.3 µM (rCysGly) to 5.4 µM (CysS). The accuracy was 95%-99%, and the repeatability and reproducibility were approximately 1.8%-3.8% and 1.9%-5.0%, respectively. An analysis of plasma samples from healthy volunteers (N = 41) showed that the mean levels of rCysGly, rCys, rGln, and CysS were 1.64, 10.6, 2.58, and 46.2 µM, respectively.
(© 2023 Wiley-VCH GmbH.)
Databáze: MEDLINE
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