title: Determination of fluoxetine and its metabolite norfluoxetine in human liver microsomes by reversed-phase HPLC in vitro.
abstract: 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.
authors.affiliation: Pharmacogenetics Research Institute, Hunan Medical University, Changsha 410078, China.
authors.forename: Z Q
authors.initials: ZQ
authors.lastname: Liu
authors.forename: Z N
authors.initials: ZN
authors.lastname: Cheng
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.forename: Z R
authors.initials: ZR
authors.lastname: Tan
authors.forename: D S
authors.initials: DS
authors.lastname: Ou-Yang
authors.forename: H H
authors.initials: HH
authors.lastname: Zhou
mesh_terms: 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
chemical_list: D005473:Fluoxetine; C036139:norfluoxetine