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2016, Número 4

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Biotecnol Apl 2016; 33 (4)


Método simple para la determinación de homocisteína y cisteína unida a proteínas en muestras de plasma humano mediante electroforesis capilar

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

Idioma: Ingles.
Referencias bibliográficas: 29
Paginas: 4301-4304
Archivo PDF: 413.69 Kb.


RESUMEN

Se desarrolló un método basado en electroforesis capilar para la detección mediante luz ultravioleta de residuos de cisteína y homocisteína unidos a proteínas en plasma humano, mediante la combinación de un paso de derivatización con apilamiento mediado por pH. Las sales y los compuestos de bajo peso molecular se eliminaron de la mezcla de proteínas mediante ultracentrifugación. A continuación, la muestra se incubó con ditiotreitol para reducir los puentes disulfuro. Los compuestos derivatizados se inyectaron a la electroforesis capilar mediante su aplicación por voltaje. El efecto de apilamiento permitió una considerable preconcentración de los analitos antes de su separación. El enfoque propuesto permite alcanzar un límite de detección de 1 µmol/L en muestras de plasma sanguíneo con el empleo de un capilar no recubierto de 48.5 cm de largo y un diámetro interno de 50 µm. El coeficiente de variación del ensayo para la homocisteína y la cisteína fue de ± 5%.


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