title: Liquid chromatography-atmospheric pressure ionization electrospray mass spectrometry determination of "hallucinogenic designer drugs" in urine of consumers.
abstract: A procedure based on liquid chromatography-mass spectrometry (LC-MS) is described for determination of 3,4-methylenedioxymethamphetamine (MDMA), 2,5-dimethoxy-4-methyl-phenethylamine (2C-D), 4-bromo-2,5-dimethoxy-beta-phenethylamine (2C-B), 1-(8-bromo-2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b'] difuran-4-yl)-2-aminoethane (2C-B-Fly), 4-ethylthio-2,5-dimethoxy-beta-phenethylamine (2C-T-2), 4-iodo-2,5-dimethoxy-beta-phenethylamine (2C-I), and 4-ethyl-2,5-dimethoxy-beta-phenethylamine (2C-E), 1-(m-chlorophenyl)piperazine (m-CPP), 4-hydroxy-N,N-diisopropyltryptamine (4-OH-DIPT) and 4-acetoxy-N,N-diisopropyltryptamine (4-acetoxy-DIPT) in urine of consumers using 3,4 methylendioxypropylamphetamine (MDPA) as internal standard. Sample preparation involved a solid-phase extraction procedure at pH 6 of both non-hydrolyzed and enzymatically hydrolyzed urine samples. Chromatography was performed on a C(18) reversed-phase column using a linear gradient of 10mM ammonium bicarbonate, pH 7.3 and acetonitrile as a mobile phase. Separated analytes were determined in LC-MS single ion monitoring mode using an atmospheric pressure ionization-electrospray ionization (ESI) interface. The assay was tested on urine samples from consumers of compounds under investigation (n=32). Limits of quantification varied between 20 and 60 ng/mL for the different analytes under investigation. Calibration curves were linear to 2000 ng/mL for all the substances under investigation, with a minimum r(2)>0.99. At three concentrations spanning the linear dynamic range of the assay, mean recoveries ranged between 55.4 and 95.6% for the different analytes. Higher analytes concentrations in hydrolyzed samples showed the presence of conjugated compounds in urine.
authors.affiliation: Department of Therapeutic Research and Medicines Evaluation, Istituto Superiore di Sanità, Rome, Italy. simona.pichini@iss.it
authors.forename: Simona
authors.initials: S
authors.lastname: Pichini
authors.forename: Mitona
authors.initials: M
authors.lastname: Pujadas
authors.forename: Emilia
authors.initials: E
authors.lastname: Marchei
authors.forename: Manuela
authors.initials: M
authors.lastname: Pellegrini
authors.forename: Jimena
authors.initials: J
authors.lastname: Fiz
authors.forename: Roberta
authors.initials: R
authors.lastname: Pacifici
authors.forename: Piergiorgio
authors.initials: P
authors.lastname: Zuccaro
authors.forename: Magi
authors.initials: M
authors.lastname: Farré
authors.forename: Rafael
authors.initials: R
authors.lastname: de la Torre
mesh_terms: D000328:Adult; D001274:Atmospheric Pressure; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D015198:Designer Drugs / Q000032:analysis* / Q000737:chemistry; D004355:Drug Stability; D005260:Female; D005615:Freezing; D006213:Hallucinogens / Q000032:analysis* / Q000652:urine; D006801:Humans; D008297:Male; D008875:Middle Aged; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D015813:Substance Abuse Detection / Q000379:methods*; D013997:Time Factors
chemical_list: D015198:Designer Drugs; D006213:Hallucinogens