Determination of estradiol metabolites in human liver microsome by high performance liquid chromatography-electrochemistry detector.
OBJECTIVE
To constitute a method to determine the estradiol metabolites in human liver microsome in low concentration of estradiol.


METHODS
Use high performance liquid chromatography after solvent extraction, evaporation, and reconstitution to separate the metabolites and use a electrochemistry detector to detect the metabolites.


RESULTS
With a mobile phase of acetic acid buffer-acetonitrile (50:50, v/v, pH 4.5) at flow rate of 1.0 mL/min and a potential of +0.7 V vs Ag/AgCl, all six composition were well separated and satisfactorily detected. There are E3, 16alpha-OHE1, 2-OHE2, E1, and two unidentified composition. The minimum detectable amount is about 100 p g on column. This method is sensitive enough to detect E1 in a substrate concentration of 1 micromol/L.


CONCLUSIONS
The method can be used to study the metabolism mechanism of estradiol in liver microsome.
Cheng
Z N
ZN
Pharmacogenetics Research Institute, Basic and Clinical Pharmacology, Hunan Medical University, Changsha 410078, China.
Huang
S L
SL
Tan
Z R
ZR
Wang
W
W
Zhou
H H
HH
D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004958:Estradiol / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism*; D004964:Estriol / Q000032:analysis*; D004970:Estrone / Q000032:analysis; D006801:Humans; D006894:Hydroxyestrones / Q000032:analysis*; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism*
D006894:Hydroxyestrones; C026702:16-hydroxyestrone; D004970:Estrone; D004958:Estradiol; C001390:2-hydroxyestradiol; D004964:Estriol