Practice in NADPH cofactor application in on-line drug biotransformation by capillary electrophoresis

ŘEMÍNEK, Roman, Marta PELCOVÁ, Zdeněk GLATZ and Wolfgang THORMANN. Practice in NADPH cofactor application in on-line drug biotransformation by capillary electrophoresis. In the 22nd International Symposium on Electro- and Liquid Phase-Separation Techniques (ITP2015). 2015.

Basic information

• Original name: Practice in NADPH cofactor application in on-line drug biotransformation by capillary electrophoresis
• Name in Czech: Použití kofaktoru NADPH v on-line studiích metabolismu léčiv pomocí kapilární elektroforézy
• Authors: ŘEMÍNEK, Roman, Marta PELCOVÁ, Zdeněk GLATZ and Wolfgang THORMANN.
• Edition: the 22nd International Symposium on Electro- and Liquid Phase-Separation Techniques (ITP2015), 2015.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• Organization unit: Faculty of Science
• Keywords (in Czech): Kapilární elektroforéza, on-line metody, metabolismus léčiv, NADPH, CYP3A4
• Keywords in English: Capillary electrophoresis, on-line methods, drug metabolism, NADPH, CYP3A4
• Tags: International impact

Abstract

Drug biotransformation performed by cytochrome P450 enzymes requires certain conditions to achieve maximal catalytic activity. The essential factor is the presence of reduced nicotinamide cofactor, NADPH (β-nicotinamide adenine dinucteotide 2´-phosphate) in sufficient amount. In general protocols NADPH regenerating system consisting of glucose 6-phosphate, glucose 6-phosphate dehydrogenase, NADP+ and MgCl2 is recommended but usage of NADPH itself simplifies the reaction mixture significantly. Fewer reagents in the mixture are more convenient for on-line enzymatic reaction performed by capillary electrophoresis (CE). However using NADPH solely evokes solving its stability issue since NADPH in aqueous solution must be available in active form for long time period in the on-line system: i) NADPH stability is lower than NADH and it is strongly dependent on pH, temperature, and ionic strength ; ii) reduced nicotinamide cofactors are particularly unstable in phosphate buffers and iii) such reagent solution must be handled with particular care.

Abstract (in Czech)

Bylo provedeno srovnání inkubačních podmínek cytochromu P450 3A4 s různými zdroji kofaktoru NADPH.

Links

• GBP206/12/G014, research and development project: Name: Centrum pokročilých bioanalytických technologií