Abstract
In forensic toxicology, amphetamine intoxications represent one of the most common case groups and present difficult questions for toxicologists. Estimating the time of consumption and the current influence of the stimulant is particularly difficult when only total amphetamine concentrations are considered. Stereoselective analysis and the consideration of metabolites can provide valuable information to facilitate interpretation. An enantioselective liquid chromatography−tandem mass spectrometry (LC-MS/MS) method for detection of amphetamine, norephedrine and 4-hydroxyamphetamine was developed. Validation showed satisfactory selectivity, sensitivity, linearity (0.5–250 ng/mL), precision and accuracy for all enantiomers. The method was applied to a collective of 425 forensic serum samples and 30 serum samples from psychiatric inpatients stating their last time of amphetamine consumption. Norephedrine and 4-hydroxyamphetamine were detected more frequently at higher amphetamine concentrations and at lower amphetamine (R)/(S) concentration ratios, possibly indicating recent consumption. Mean (R)/(S) ratio of amphetamine was 1.14, whereas higher ratios (mean 1.36) were found for amphetamine concentrations below 100 ng/mL. The (R)/(S) ratios of psychiatric inpatients significantly correlated with the reported time intervals to last consumption. The use of amphetamine (R)/(S) ratios and the simultaneous detection of metabolites are promising factors that can facilitate estimation of consumption time and current impairment.
Highlights
Amphetamine remains one of the most frequently abused drugs worldwide [1]
Precision (relative standard deviation (RSD)) and bias data at the lowest calibration level (0.5 ng/mL) were within the acceptable limits of the guideline, so this was accepted as the lower limit of quantification (LLOQ)
For enantiomers of 4-hydroxyamphetamine and norephedrine, the 0.5 ng/mL level was the limit of detection (LOD); for amphetamine enantiomers, 0.1 ng/mL was accepted as LOD, only quantifiable results (≥0.5 ng/mL) were considered for this study
Summary
Amphetamine remains one of the most frequently abused drugs worldwide [1]. Intoxications with this stimulant represent one of the most common case groups and present difficult questions for toxicologists [2,3,4].Amphetamine is a sympathomimetic amine that increases brain levels of norepinephrine, serotonin and dopamine by promoting release and inhibiting reuptake of these neurotransmitters [5,6]. Amphetamine remains one of the most frequently abused drugs worldwide [1] Intoxications with this stimulant represent one of the most common case groups and present difficult questions for toxicologists [2,3,4]. The determination of the absolute serum amphetamine concentration hardly allows a statement on the time of consumption and on the current influence of the stimulant [15]. This is due to differences in quantity and frequency of dosing due to habituation effects [16], as well as interindividual differences in metabolism [17,18]
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