Application of liquid chromatography-electrospray ionization mass spectrometry for study of steroid-converting enzymes.
A high-performance liquid chromatography-atmospheric pressure ionization-electrospray ionization mass spectrometry (HPLC-API-ESI-MS) method was developed for the analysis of steroids in a study of steroid-converting enzymes. Separations ware done on a Zorbax Eclipse XDB-C18 column (eluted with a linear methanol-water-acetic acid gradient) and identification of the steroids involved was done by API-ESI-MS using positive ion mode and extracted ion analysis. The applicability of the present method for studying steroid metabolism was proven in assaying two steroid-converting enzymes (20beta-hydroxysteroid dehydrogenase and 11beta-hydroxysteroid dehydrogenase) in various biological samples (rat and chicken intestine, chicken oviduct).
Miksík
Ivan
I
Institute of Physiology, Academy of Sciences of the Czech Republic, Vídenská 1083, CZ 142 20 Prague 4, Czech Republic. miksik@biomed.cas.cz
Mikulíková
Katerina
K
Pácha
Jirí
J
Kucka
Marek
M
Deyl
Zdenek
Z
D042842:11-beta-Hydroxysteroid Dehydrogenases / Q000032:analysis; D000818:Animals; D002138:Calibration; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D003345:Corticosterone / Q000378:metabolism; D004798:Enzymes / Q000737:chemistry*; D005260:Female; D006168:Guinea Pigs; D007202:Indicators and Reagents; D007422:Intestines / Q000201:enzymology; D008297:Male; D010057:Oviducts / Q000201:enzymology; D051381:Rats; D017208:Rats, Wistar; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet; D013256:Steroids / Q000378:metabolism*
D004798:Enzymes; D007202:Indicators and Reagents; D013256:Steroids; D042842:11-beta-Hydroxysteroid Dehydrogenases; D003345:Corticosterone