Acidic Dithiothreitol Reduction on Disulfide Bonds using Cation‐Exchange Chromatography

Autor:	Scott Whitman, Jinmo Huang
Rok vydání:	2006
Předmět:	chemistry.chemical_classification Chromatography Ion exchange Clinical Biochemistry Ion chromatography Cystine Pharmaceutical Science Biochemistry High-performance liquid chromatography Dithiothreitol Analytical Chemistry chemistry.chemical_compound chemistry Ninhydrin Thiol Cysteine
Zdroj:	Journal of Liquid Chromatography & Related Technologies. 29:2513-2520
ISSN:	1520-572X 1082-6076
DOI:	10.1080/10826070600914984
Popis:	Dithiothreitol has been utilized to reduce the disulfide bonds between homocysteine and other thiols in total plasma homocysteine determination. The completeness of the reduction of disulfides into thiols is a prerequisite for accurate quantitation of total plasma homocysteine. In order to evaluate the effectiveness of dithiothreitol reduction on disulfide bonds, the acidic mixture of cystine and homocystine is reduced under controlled parameters including dithiothreitol concentration, reduction temperature, and reaction time. The reaction mixture is analyzed by cation‐exchange chromatography. It is concluded that 1.0 M dithiothreitol concentration, 70°C reduction temperature, and 1 hour reaction time are required to convert more than 96% of the disulfides of cystine and homocystine to their thiols in acidic medium. This study also proves that dithiothreitol is able to keep cysteine and homocysteine from being reoxidized back to their disulfides.
Databáze:	OpenAIRE
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