Determination of fluoxetine and its metabolite norfluoxetine in human liver microsomes by reversed-phase HPLC in vitro.
OBJECTIVE
A high-performance liquid chromatography (HPLC) method was developed for the determination of fluoxetine (FLU) and its metabolite norfluoxetine (N-FLU) in human liver microsomes in vitro.


METHODS
An incubation buffer containing human liver microsomes, NADPH-generating system, and FLU, after termination of enzyme reaction and addition of nortriptyline (NOR) as internal standard (IS), was extracted with n-hexane/acetonitrile, and separated on a reversed-phase ODS column. Detection was achieved at 226 nm by ultraviolet detector (UV).


RESULTS
The limit of detection was 5 micrograms/L for both FLU and N-FLU. No potential interference was found. The method provides recoveries of up to 94%-104% and acceptable coefficients of variation were found for both within-run (< 7.8%) and day to day (< 9.1%) assays.


CONCLUSIONS
This method is rapid, sensitive, and simple for studying the metabolism of FLU and N-FLU.
Liu
Z Q
ZQ
Pharmacogenetics Research Institute, Hunan Medical University, Changsha 410078, China.
Cheng
Z N
ZN
Wang
W
W
Tan
Z R
ZR
Ou-Yang
D S
DS
Zhou
H H
HH
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005473:Fluoxetine / Q000031:analogs & derivatives / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008862:Microsomes, Liver / Q000737:chemistry* / Q000378:metabolism
D005473:Fluoxetine; C036139:norfluoxetine