Problems of Forensic Sciences 2006 Vol. 66 (LXVI) 131-139
DETERMINATION OF TRAZODONE AND ITS METABOLITE 1-(3-CHLOROPHENYL)PIPERAZINE (mCPP) IN HAIR BY THE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHOD COUPLED WITH MASS SPECTROMETRY (HPLC-ESI-MS)
Roman STANASZEK, Wojciech LECHOWICZ
Institute of Forensic Sciences, Krakow, Poland
Streszczenie
The aim of this paper was to develop, optimise and validate an analytical procedure for determining the popular antidepressant drug trazodone and its active metabolite 1-(3-chlorophenyl)piperazine (mCPP) in hair. mCPP, being a derivative of piperazine, is considered to be a so-called designer drug. It exhibits stimulating and hallucinogenic activity similar to MDMA. Hair samples (50 mg) were extracted by 1-chlorobutane after prior alkaline hydrolysis (0.5 M NaOH, 1 h, 70°C), and the obtained extracts were analysed by high performance liquid chromatography coupled with electrospray ionisation mass spectrometry (HPLC-ESI-MS). Applying the technique of direct introduction of the sample into the MS, optimisation of ESI-MS parameters was carried out and the following values were obtained: capillary voltage 3 kV, fragmentor voltage 45 V, ion source temperature 100°C, desolvation temperature 300°C, and desolvation gas flow 600 l/h. Quantitative analysis was carried out in the selected ion monitoring (recording) mode (SIR). Quantitative analysis was carried out on the basis of analytical signals obtained for positive pseudo-molecular ions of the determined substances (m/z – 197 for mCPP, m/z – 372 for trazodone) and deuterated ketamine (m/z – 242), which was used as an internal standard. Additionally, 2 ions each were monitored for analytes: m/z 199, 154 for mCPP and 374, 176 for trazodone, which were used for identification together with pseudomolecular ions. For mCPP, the limit of detection was 0.1 ng/mg of hair, and for trazodone it was 0.02 ng/mg. The linearity for both compounds was confirmed in the range 0 to 5 ng/mg. The obtained values of other validation parameters fulfilled criteria required for this type of analysis and were: intra-group precision for a concentration of 0.5 ng/mg: mCPP – CV 6.5%, trazodone – CV 14.0%; for a concentration of 2 ng/mg: mCPP – CV 5.3% and CV 10.1% for trazodone; recovery: 70% for mCPP and 129% for trazodone at a concentration of 2 ng/mg. The developed method was applied to determination of trazodone and mCPP in 7 authentic samples of hair. In the case of a hair sample taken from a person one month after the person had consumed a single dose of 75 mg trazodone, the following concentrations were ascertained: segment 1 (1 cm) trazodone – 5.93 ng/mg and mCPP – 0.1 ng/mg; segment 2 (1 cm) trazodone – 0.04 ng/mg, mCPP not detected. The six remaining samples were negative. The developed method may be applied to differentiating between consumption of the hallucinogenic mCPP and consumption of its precursor trazodone.
Słowa kluczowe
Trazodone; mCPP; Hair; LC-ESI-MS.