Problems of Forensic Sciences 2007 Vol. 69 (LXIX) 90-97
DETERMINATION OF DESIPRAMINE AND NORTRIPTYLINE IN BLOOD BY MEANS OF THE HPLC-DAD METHOD USING 7,7,8,8-TETRACYANOQUINODIMETHANE (TCNQ) AS A DERIVATISATION AGENT
Michał WOŹNIAKIEWICZ1, Jagoda KUCZARA1, Paweł KOŚCIELNIAK1,2
1Faculty of Chemistry, Jagiellonian University, Krakow
2Institute of Forensic Research, Krakow
Streszczenie
The aim of present study was to develop a sensitive and reliable method of simultaneous determination of desipramine and nortriptyline in blood by means of high-performance liquid chromatography coupled with a diode array spectrophotometer (HPLC-DAD). For this purpose, a procedure based on formation of colour derivatives of these drugs with 7,7,8,8-tetracyanoquinodimethane (TCNQ) was proposed. The tested drugs and the internal standard were added to blood samples, and then they were extracted by the microwave assisted liquid-liquid extraction method. The separated organic layer was evaporated and was subjected to derivatisation by TCNQ reagent dissolved in acetonitrile. Method validation was performed and the limits of detection (0.03–0.05 μg/ml) and quantitation (0.09–0.18 μg/ml) of both analytes were determined, as were the precision and accuracy of their determination at two concentration levels. The worked-out method allows determination of desipramine and nortriptyline present in blood at therapeutic levels.
Słowa kluczowe
Desipramine; Nortriptyline; TCNQ; HPLC-DAD; Derivatisation.