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Determination of estradiol metabolites in human liver microsome by high performance liquid
Z N Cheng1, S L Huang, Z R Tan
1Pharmacogenetics Research Institute, Basic and Clinical Pharmacology, Hunan Medical University, Changsha 410078, China.
Aim:
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.
Conclusion:
The method can be used to study the metabolism mechanism of estradiol in liver microsome.
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