Summary
An HPLC-UV method for the simultaneous identification of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA) in urine is described. It includes a rapid extraction procedure of the 4 analogs from urine using Extrelut 3 columns, derivatization with sodium 1,2-naphthoquinone-4-sulphonate (NQS) to obtain highly chromophoric UV-VIS derivatives, and a final HPLC analysis using an ion-pair reversed-phase technique with eluent monitoring at 480 nm. Structural characterization of the derivatives obtained by mass spectrometry is reported. Recoveries of the amphetamines were in the range 80–85% at concentrations of 300 ng/ml. Practical detection limits were 40–60 ng/ml (S/N ratio = 10) for all derivatives. The chromatographic peaks of the NQS derivatized amphetamines are fairly narrow and well resolved. The method is simple, rapid, quite sensitive, and specific for convenient confirmation of preliminary positive results obtained with immunoassays.
Zusammenfassung
Eine HPLC-UV-Methode zur gleichzeitigen Bestimmung von Amphetamin, Methamphetamin, 3,4-Methylendioxyamphetamin (MDA) und 3,4-Methylendioxymethamphetamin (MDMA) im Urin wird beschrieben. Vorgestellt wird eine Schnellextraktion der 4 Amphetamine über Extrelut-3-Säulen und eine Derivatisierung mit Natrium-1,2-naphthochinon-4-sulfonat (NQS) um hochchromophore UV-VIS-Derivate zu erhalten. Abschließend erfolgt eine HPLC-Analyse mittels einer reversed phase Technik (Ionenpaar) mit Detektion des Eluats bei 480 nm. Die Struktur der NQS-Derivate wird durch Massenspektrometrie charakterisiert. Die Wiederfindungsraten der Amphetamine liegen bei Konzentrationen von 300 ng/ml bei 80–85%. Die Nachweisgrenze liegt für alle Derivate bei 40–60 ng/ml (S/N-Verhältnis = 10). Die Trennung der NQS-derivatisierten Amphetamine gelingt gut, die Methode ist einfach, schnell, empfindlich und spezifisch genug für die Bestätigung vorläufiger Screening-Resultate, wie sie mit Hilfe von Immunoassays erhalten werden.
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Tedeschi, L., Frison, G., Castagna, F. et al. Simultaneous identification of amphetamine and its derivatives in urine using HPLC-UV. Int J Leg Med 105, 265–269 (1993). https://doi.org/10.1007/BF01370383
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DOI: https://doi.org/10.1007/BF01370383