Sensitive HPLC method for the determination of fluoxetine and norfluoxetine in human plasma with fluorescence detection


Erturk S., Cetin S., Atmaca S., Ersoy L., Baktir G.

THERAPEUTIC DRUG MONITORING, cilt.27, sa.1, ss.38-43, 2005 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 27 Sayı: 1
  • Basım Tarihi: 2005
  • Doi Numarası: 10.1097/00007691-200502000-00009
  • Dergi Adı: THERAPEUTIC DRUG MONITORING
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.38-43
  • İstanbul Üniversitesi Adresli: Evet

Özet

A selective, sensitive, and precise HPLC method for the simultaneous determination of fluoxetine (FL) and its N-demethylated metabolite norfluoxetine (NFL) in human plasma has been developed. Following extraction with n-hexane, FL, NFL, and fluvoxamine (internal standard) were derivatized with 7-chloro-4-nitrobenzofurazan (NBD-Cl) under weakly alkaline conditions. NBD derivatives were extracted with chloroform after acidification and chromatographed on a reversed-pbase column with gradient elution using acetonitrile and 0.1 mol/L nitric acid (pH 3) solution. Calibration curves were linear over the range of 1.0-100.0 ng/mL and 0.1-50.0 ng/mL for FL and NFL, respectively, with inter- and intraassay precision given by a relative standard deviation (RSD%) of less than 9.2%. The lower limits of quantification were 1.0 ng/mL for FL and 0.1 ng/mL for NFL. Recoveries of FL and NFL from plasma at three different concentrations were assessed. Average recovery was about 100% for both substances. The assay was applied to pharmacokinetic study in 2 healthy volunteers after a single oral administration of 40 mg of FL.