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

a 2015

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

ŘEMÍNEK, Roman, Marta PELCOVÁ, Zdeněk GLATZ a Wolfgang THORMANN

Základní údaje

Originální název

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

Název česky

Použití kofaktoru NADPH v on-line studiích metabolismu léčiv pomocí kapilární elektroforézy

Autoři

ŘEMÍNEK, Roman, Marta PELCOVÁ, Zdeněk GLATZ a Wolfgang THORMANN

Vydání

the 22nd International Symposium on Electro- and Liquid Phase-Separation Techniques (ITP2015), 2015

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10600 1.6 Biological sciences

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova česky

Kapilární elektroforéza, on-line metody, metabolismus léčiv, NADPH, CYP3A4

Klíčová slova anglicky

Capillary electrophoresis, on-line methods, drug metabolism, NADPH, CYP3A4

Příznaky

Mezinárodní význam

Změněno: 31. 1. 2017 09:30, Ing. Mgr. Roman Řemínek, Ph.D.

Anotace

ORIG CZ

V originále

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.

Česky

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

Návaznosti

GBP206/12/G014, projekt VaV

Název: Centrum pokročilých bioanalytických technologií

Citovat

ŘEMÍNEK, Roman, Marta PELCOVÁ, Zdeněk GLATZ a 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.

@proceedings{1370827,
   author = {Řemínek, Roman and Pelcová, Marta and Glatz, Zdeněk and Thormann, Wolfgang},
   booktitle = {the 22nd International Symposium on Electro- and Liquid Phase-Separation Techniques (ITP2015)},
   keywords = {Capillary electrophoresis, on-line methods, drug metabolism, NADPH, CYP3A4},
   language = {eng},
   title = {Practice in NADPH cofactor application in on-line drug biotransformation by capillary electrophoresis},
   year = {2015}
}

TY  - CONF
ID  - 1370827
AU  - Řemínek, Roman - Pelcová, Marta - Glatz, Zdeněk - Thormann, Wolfgang
PY  - 2015
TI  - Practice in NADPH cofactor application in on-line drug biotransformation by capillary electrophoresis
KW  - Capillary electrophoresis, on-line methods, drug metabolism, NADPH, CYP3A4
N2  - 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.
ER  -

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

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