Abstract
Synthetic cathinones are novel stimulants derived from cathinone, with amphetamines or cocaine-like effects, often labeled “not for human consumption” and considered “legal highs”. Emergence of these new designer drugs complicate interpretation of forensic and clinical cases, with introduction of many new analogs designed to circumvent legislation and vary effects and potencies. We developed a method for the simultaneous quantification of 28 synthetic cathinones, including four metabolites, in urine by liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). These cathinones include cathinone, methcathinone, and synthetic cathinones position-3’-substituted, N-alkyl-substituted, ring-substituted, methylenedioxy-substituted, and pyrrolidinyl-substituted. One mL phosphate buffer pH 6 and 25 μL IStd solution were combined with 0.25 mL urine, and subjected to solid phase cation exchange extraction (SOLA SCX). The chromatographic reverse-phase separation was achieved with a gradient mobile phase of 0.1 % formic acid in water and in acetonitrile in 20 min. We employed a Q Exactive high resolution mass spectrometer, with compounds identified and quantified by target-MSMS experiments. The assay was linear from 0.5–1 to 100 μg/L, with limits of detection of 0.25–1 μg/L. Imprecision (n = 20) was <15.9 % and accuracy (n = 20) 85.2–118.1 %. Extraction efficiency was 78.9–116.7 % (CV 1.4–16.7 %, n = 5), process efficiency 57.7–104.9 %, and matrix effects from −29.5 % to 1.5 % (CV 1.9–13.1 %, n = 10). Most synthetic cathinones were stable at 4 °C for 72 h (n = 27) and after 3 freeze-thaw cycles (n = 26), but many (n = 19) were not stable at room temperature for 24 h (losses up to −67.6 %). The method was applied to authentic urine specimens from synthetic cathinone users. This method provides a comprehensive confirmation method for 28 synthetic cathinones in urine, with good selectivity and specificity.
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References
Kelly JP (2011) Cathinone derivatives: a review of their chemistry, pharmacology and toxicology. Drug Test Anal 3(7–8):439–453
Strano-Rossi S, Cadwallader AB, de la Torre X, Botre F (2010) Toxicological determination and in vitro metabolism of the designer drug methylenedioxypyrovalerone (MDPV) by gas chromatography/mass spectrometry and liquid chromatography/quadrupole time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 24(18):2706–2714
United Nations (1971) Covention on psychotropics substances. http://www.unodc.org/pdf/convention_1971_en.pdf. Accessed 6 August 2013
Drug Enforcement Administrations (2013) Schedules of controlled substances: placement of methylone into schedule I. Fed Regist 78(71):21818–21825
Drug Enforcement Administrations (2012) Drug Abuse Prevention Act 2012 1152b:3187–3139
European Monitoring Centre for Drugs and Drug Addiction (2013) Drug profiles: synthetic cathinones. http://www.emcdda.europa.eu/publications/drug-profiles/synthetic-cathinones. Accessed 6 August 2013
Gunderson EW, Kirkpatrick MG, Willing LM, Holstege CP (2013) Substituted cathinone products: a new trend in “bath salts” and other designer stimulant drug use. J Addict Med 7(3):153–162
Winstock AR, Mitcheson LR, Deluca P, Davey Z, Corazza O, Schifano F (2011) Mephedrone, new kid for the chop? Addiction 106(1):154–161
Kriikku P, Wilhelm L, Schwarz O, Rintatalo J (2011) New designer drug of abuse: 3,4-Methylenedioxypyrovalerone (MDPV). Findings from apprehended drivers in Finland. Forensic Sci Int 210(1–3):195–200
American Association of Poison Control Centers. Alerts: bath salts. http://www.aapcc.org/alerts/bath-salts/. Accessed 6 August 2013
Prosser JM, Nelson LS (2012) The toxicology of bath salts: a review of synthetic cathinones. J Med Toxicol 8(1):33–42
Meyer MR, Wilhelm J, Peters FT, Maurer HH (2010) Beta-keto amphetamines: studies on the metabolism of the designer drug mephedrone and toxicological detection of mephedrone, butylone, and methylone in urine using gas chromatography–mass spectrometry. Anal Bioanal Chem 397(3):1225–1233
Pedersen AJ, Reitzel LA, Johansen SS, Linnet K (2013) In vitro metabolism studies on mephedrone and analysis of forensic cases. Drug Test Anal 5(6):430–438
Meyer MR, Du P, Schuster F, Maurer HH (2010) Studies on the metabolism of the α-pyrrolidinophenone designer drug methylenedioxy-pyrovalerone (MDPV) in rat and human urine and human liver microsomes using GC–MS and LC–high-resolution MS and its detectability in urine by GC–MS. J Mass Spectrom 45(12):1426–1442
Kamata HT, Shima N, Zaitsu K, Kamata T, Miki A, Nishikawa M, Katagi M, Tsuchihashi H (2006) Metabolism of the recently encountered designer drug, methylone, in humans and rats. Xenobiotica 36(8):709–723
Zaitsu K, Katagi M, Kamata HT, Kamata T, Shima N, Miki A, Tsuchihashi H, Mori Y (2009) Determination of the metabolites of the new designer drugs bk-MBDB and bk-MDEA in human urine. Forensic Sci Int 188(1–3):131–139
Shima N, Katagi M, Kamata H, Matsuta S, Nakanishi K, Zaitsu K, Kamata T, Nishioka H, Miki A, Tatsuno M, Sato T, Tsuchihashi H, Suzuki K (2013) Urinary excretion and metabolism of the newly encountered designer drug 3,4-dimethylmethcathinone in humans. Forensic Toxicol 31(1):101–112
Tyrkko E, Pelander A, Ketola RA, Ojanpera I (2013) In silico and in vitro metabolism studies support identification of designer drugs in human urine by liquid chromatography/quadrupole-time-of-flight mass spectrometry. Anal Bioanal Chem. doi:10.1007/s00216-013-7137-1
O’Byrne PM, Kavanagh PV, McNamara SM, Stokes SM (2013) Screening of stimulants including designer drugs in urine using a liquid chromatography tandem mass spectrometry system. J Anal Toxicol 37(2):64–73
Ojanpera IA, Heikman PK, Rasanen IJ (2011) Urine analysis of 3,4-methylenedioxypyrovalerone in opioid-dependent patients by gas chromatography–mass spectrometry. Ther Drug Monit 33(2):257–263
Thornton SL, Gerona RR, Tomaszewski CA (2012) Psychosis from a bath salt product containing flephedrone and MDPV with serum, urine, and product quantification. J Med Toxicol 8(3):310–313
Torrance H, Cooper G (2010) The detection of mephedrone (4-methylmethcathinone) in 4 fatalities in Scotland. Forensic Sci Int 202(1–3):e62–e63
Grueninger D, Englert R (2011) Determination of the amphetamine-like designer drugs methcathinone and 4-methylmethcathinone in urine by LC-MS/MS. Ann Toxicol Anal 23(1):7–14
Paul BD, Cole KA (2001) Cathinone (Khat) and methcathinone (CAT) in urine specimens: a gas chromatographic-mass spectrometric detection procedure. J Anal Toxicol 25:525–530
Marinetti LJ, Antonides HM (2013) Analysis of synthetic cathinones commonly found in bath salts in human performance and postmortem toxicology: method development, drug distribution and interpretation of results. J Anal Toxicol 37(3):135–146
Bell C, George C, Kicman AT, Traynor A (2011) Development of a rapid LC-MS/MS method for direct urinalysis of designer drugs. Drug Test Anal 3(7–8):496–504
Meyer MR, Prosser D, Maurer HH (2013) Studies on the metabolism and detectability of the designer drug beta-naphyrone in rat urine using GC-MS and LC-HR-MS/MS. Drug Test Anal 5(4):259–265
Meyer MR, Vollmar C, Schwaninger AE, Wolf E, Maurer HH (2012) New cathinone-derived designer drugs 3-bromomethcathinone and 3-fluoromethcathinone: studies on their metabolism in rat urine and human liver microsomes using GC–MS and LC–high-resolution MS and their detectability in urine. J Mass Spectrom 47(2):253–262
Krouwer JS, Rabinowitz R (1984) How to improve estimates of imprecision. Clin Chem 30(2):290–292
Chesher D (2008) Evaluating assay precision. Clin Biochem Rev 29(Suppl 1):S23–S26
Springer D, Peters FT, Fritschi G, Maurer HH (2003) New designer drug 4’-methyl-alpha-pyrrolidinohexanophenone: studies on its metabolism and toxicological detection in urine using gas chromatography–mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 789(1):79–91
Tsujikawa K, Mikuma T, Kuwayama K, Miyaguchi H, Kanamori T, Iwata YT, Inoue H (2012) Degradation pathways of 4-methylmethcathinone in alkaline solution and stability of methcathinone analogs in various pH solutions. Forensic Sci Int 220(1–3):103–110
Johnson RD, Botch-Jones SR (2013) The stability of four designer drugs: MDPV, mephedrone, BZP and TFMPP in three biological matrices under various storage conditions. J Anal Toxicol 37(2):51–55
Sorensen LK (2011) Determination of cathinones and related ephedrines in forensic whole-blood samples by liquid-chromatography-electrospray tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 879(11–12):727–736
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This research was supported by the Intramural Research Program of the National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH).
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Concheiro, M., Anizan, S., Ellefsen, K. et al. Simultaneous quantification of 28 synthetic cathinones and metabolites in urine by liquid chromatography-high resolution mass spectrometry. Anal Bioanal Chem 405, 9437–9448 (2013). https://doi.org/10.1007/s00216-013-7386-z
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DOI: https://doi.org/10.1007/s00216-013-7386-z