Determination of D-fenfluramine, D-norfenfluramine and fluoxetine in plasma, brain tissue and brain microdialysate using high-performance liquid chromatography after precolumn derivatization with dansyl chloride.
A HPLC method is described for the simultaneous determination of D-fenfluramine (FEN), D-norfenfluramine (NF) and fluoxetine (FLX) using fluorometric detection after precolumn derivatization with dansyl-chloride. The method has limits of quantitation of 200 fmol for FEN and NF, 500 fmol for FLX in brain microdialysate, and 1 pmol for NF and FEN, and 2 pmol for FLX in plasma. Brain tissue standards were linear between 5 and 200 pmol/mg for all three compounds. The inter-assay variability (relative standard deviation) was 6.6%, 6.9% and 9.3% for FEN, 4.6%, 3.7% and 7.9% for NF and 10.4%, 4.9% and 12.2% for FLX, for brain microdialysate (2 pmol/microl), plasma (2 pmol/ microl) and brain tissue (50 pmol/mg), respectively. Intra-assay variability was always lower, typically several times lower than inter-assay variability. Extraction recovery was 108% and 48% for FEN, 105% and 78% for NF and 94% and 45% for FLX, in plasma (2 pmol/microl) and brain tissue (5 pmol/mg), respectively. Due to the stability of the dansyl-chloride derivatives this method is well suited for an autoinjector after manual derivatization with dansyl chloride at room temperature for 4 h.
Clausing
P
P
Division of Neurotoxicology, National Center for Toxicological Research, Jefferson, AR 72079-9502, USA.
Rushing
L G
LG
Newport
G D
GD
Bowyer
J F
JF
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003619:Dansyl Compounds / Q000737:chemistry*; D005277:Fenfluramine / Q000097:blood / Q000378:metabolism*; D005473:Fluoxetine / Q000097:blood / Q000378:metabolism*; D007202:Indicators and Reagents; D017551:Microdialysis; D009642:Norfenfluramine / Q000097:blood / Q000378:metabolism*; D051381:Rats; D012015:Reference Standards; D017367:Selective Serotonin Reuptake Inhibitors / Q000097:blood / Q000378:metabolism*
D003619:Dansyl Compounds; D005277:Fenfluramine; D005473:Fluoxetine; D007202:Indicators and Reagents; D009642:Norfenfluramine; D017367:Selective Serotonin Reuptake Inhibitors; C029038:dansyl chloride