化学
色谱法
毛细管电泳
等电聚焦
重复性
检出限
磷酸
聚乙二醇
胱氨酸
半胱氨酸
生物化学
有机化学
酶
作者
Alexander Ivanov,Mikhail А. Popov,В. В. Александрин,Polina Alexandrovna Pudova,Maria Pavlovna Galdobina,Arkady Andreevich Metelkin,Maria Petrovna Kruglova,Roman Maslennikov,Ekaterina Silina,Victor Stupin,Аслан Амирханович Кубатиев
标识
DOI:10.1002/elps.202300196
摘要
Abstract 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.
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