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2016, Number 4

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Biotecnol Apl 2016; 33 (4)

simple method for determining protein-bound homocysteine and cysteine in human plasma by capillary electrophoresis

Danielevich VE, Vladimirovich IA, Iusupovna KM, Petrovich LB, Amirkhanovich KA

Language: English
References: 29
Page: 4301-4304
PDF size: 413.69 Kb.


ABSTRACT

A capillary electrophoresis method with UV detection has been developed for the determination of protein-bound cysteine and homocysteine in human plasma based on the combination of derivatization step with pH-mediated base stacking. Centrifugal ultrafiltration was carried out to clarify plasma proteins from salts and low-molecular weight compounds. Thereafter, the sample was incubated with dithiothreitol to reduce the disulfides and release protein-bound aminothiols. The released thiols were derivatized with thiocarbonyldiimidazole and injected into the capillary electrophoresis by voltage. Due to the stacking effects it is possible to perform a considerable on-line pre-concentration of the analytes. The proposed approach allows to reach a detection limit of 1 µmol/L in blood plasma using 48.5 cm total length 50-µm i.d. uncoated capillary. The coefficient of variation of the assay was within ± 5 % for both homocysteine and cysteine.


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