Abstract
Methamphetamine (MA) and amphetamine (AM) are central nervous system doping agents, listed in the WADA Prohibited List, which in the past years showed the highest prevalence among all stimulants. Specifically, concerning these stimulants, it is essential to distinguish between d and l-isomers of AM and MA due to their distinction in illicit or legal nonprescription use. The main separation methods used for the urinary determination of MA and AM enantiomers are based on chromatographic procedures, such as gas chromatography after chiral derivatization or liquid chromatography (LC) applying chiral columns, followed by the mass spectrometer (MS) detection. The use of the optically pure reagent R-(-)-α-methoxy-α-(trifluoromethyl) phenylacetic acid chloride (R-MTPA-Cl – Mosher’s Reagent) allows the isomeric separation by both chromatographic procedures while preserving the isomeric composition of the substances. Therefore, in the present work, an LC-MS2 approach has been developed for AM and MA detection in urine after Mosher derivatization. The method was validated and can be used for the enantioselective confirmation of AM and MA for doping control purposes and forensic analysis in general, being useful for the distinction between the medicinal and illicit use of both stimulants.
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References
World Anti-Doping Agency (WADA) (2017) Anti-doping testing figures. WADA, Montreal
Wang T, Shen B, Shi Y, Xiang P, Yu Z (2015) Chiral separation and determination of R/S-methamphetamine and its metabolite R/S-amphetamine in urine using LC-MS/MS. Forensic Sci Int 246:72–78. https://doi.org/10.1016/j.forsciint.2014.11.009
World Anti-Doping Agency (2020) The prohibited list. WADA, Montreal
Holler JM, Vorce SP, Bosy TZ, Jacobs A (2005) Quantitative and isomeric determination of amphetamine and methamphetamine from urine using a nonprotic elution solvent and R(-)-α-methoxy-α- trifluoromethylphenylacetic acid chloride derivatization. J Anal Toxicol 29:652–657. https://doi.org/10.1093/jat/29.7.652
Pozo OJ, Lootens L, Van Eenoo P, Deventer K, Meuleman P, Leroux-Roels G, Parr MK, Schänzer W, Delbeke FT (2009) Combination of liquid-chromatography tandem mass spectrometry in different scan modes with human and chimeric mouse urine for the study of steroid metabolism. Drug Metab Pharmacokinet 55:139–150.
Badoud F, Guillarme D, Boccard J, Grata E, Saugy M, Rudaz S, Veuthey JL (2011) Analytical aspects in doping control: Challenges and perspectives. Forensic Sci Int 213:49–61. https://doi.org/10.1016/j.forsciint.2011.07.024
Maas A, Losacker M, Hess C (2018) Chromatographic separation of R/S-enantiomers of amphetamine and methamphetamine: pathways of methamphetamine synthesis and detection in blood samples by qualitative enantioselective LC–MS/MS analysis. Forensic Sci Int 291:138–143. https://doi.org/10.1016/j.forsciint.2018.08.013
Hegstad S, Havnen H, Helland A, Spigset O, Frost J (2018) Enantiomeric separation and quantification of R/S-amphetamine in urine by ultra-high performance supercritical fluid chromatography tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 1077–1078:7–12. https://doi.org/10.1016/j.jchromb.2018.01.028
Concheiro M, Simões SMSS, Quintela Ó, Castro A, Dias MJR, Cruz A, López-Rivadulla M (2007) Fast LC-MS/MS method for the determination of amphetamine, methamphetamine, MDA, MDMA, MDEA, MBDB and PMA in urine. Forensic Sci Int 171:44–51. https://doi.org/10.1016/j.forsciint.2006.10.004
Chou CC, Lee MR (2005) Solid phase microextraction with liquid chromatography-electrospray ionization-tandem mass spectrometry for analysis of amphetamine and methamphetamine in serum. Anal Chim Acta 538:49–56. https://doi.org/10.1016/j.aca.2005.02.018
Fuh MR, Wu TY, Lin TY (2006) Determination of amphetamine and methamphetamine in urine by solid phase extraction and ion-pair liquid chromatography-electrospray-tandem mass spectrometry. Talanta 68:987–991. https://doi.org/10.1016/j.talanta.2005.06.057
Strano-Rossi S, Botrè F, Bermejo AM, Tabernero MJ (2009) A rapid method for the extraction, enantiomeric separation and quantification of amphetamines in hair. Forensic Sci Int 193:95–100. https://doi.org/10.1016/j.forsciint.2009.09.016
Weiß JA, Kadkhodaei K, Schmid MG (2017) Indirect chiral separation of 8 novel amphetamine derivatives as potential new psychoactive compounds by GC–MS and HPLC. Sci Justice 57:6–12. https://doi.org/10.1016/j.scijus.2016.08.007
Popovic A, McBriar T, He P, Beavis A (2017) Chiral determination and assay of optical isomers in clandestine drug laboratory samples using LC-MSMS. Anal Methods 9:3380–3387. https://doi.org/10.1039/c6ay03125k
Ali I, Sahoo DR, ALOthman ZA, Alwarthan AA, Asnin L, Larsson, B, (2015) Validated chiral high performance liquid chromatography separation method and simulation studies of dipeptides on amylose chiral column. J Chromatogr A 1406:201–209. https://doi.org/10.1016/j.chroma.2015.06.027
Ali I, Suhail M, ALOthman ZA, Al-Mohaimeed AM, Alwarthan A, (2020) Chiral resolution of four stereomers and simulation studies of newly synthesized antibacterial agents having two chiral centers. Sep Purif Technol 236:116256. https://doi.org/10.1016/j.seppur.2019.116256
Al-Othman ZA, Ali I (2012) Rapid and economic chiral-HPLC method of nebivolol enantiomers resolution in dosage formulation. Biomed Chromatogr 26:775–780. https://doi.org/10.1002/bmc.1728
Mazzarino M, Fiacco I, de la Torre X, Botrè F (2011) Screening and confirmation analysis of stimulants, narcotics and beta-adrenergic agents in human urine by hydrophilic interaction liquid chromatography coupled to mass spectrometry. J Chromatogr A 1218:8156–8167. https://doi.org/10.1016/j.chroma.2011.09.020
Haneef J, Shaharyar M, Husain A, Rashid M, Mishra R, Parveen S, Ahmed N, Pal M, Kumar D (2013) Application of LC-MS/MS for quantitative analysis of glucocorticoids and stimulants in biological fluids. J Pharm Anal 3:341–348. https://doi.org/10.1016/j.jpha.2013.03.005
Reddy I, Beotra A, Jain S, Ahi S (2009) A simple and rapid ESI-LC-MS/MS method for simultaneous screening of doping agents in urine samples. Indian J Pharmacol 41:80–86. https://doi.org/10.4103/0253-7613.51347
Thevis M, Thomas A, Schänzer W (2011) Current role of LC-MS(/MS) in doping control. Anal Bioanal Chem 401:405–420. https://doi.org/10.1007/s00216-011-4859-9
Wood M, De Boeck G, Samyn N, Morris M, Cooper DP, Maes RAA, De Bruijn EA (2003) Development of a rapid and sensitive method for the quantitation of amphetamines in human plasma and oral fluid by LC-MS-MS. J Anal Toxicol 27:78–87. https://doi.org/10.1093/jat/27.2.78
Nakanishi K, Katagi M, Zaitsu K, Shima N, Kamata H, Miki A, Kato H, Harada KI, Tsuchihashi H, Suzuki K (2012) Simultaneous enantiomeric determination of MDMA and its phase I and phase II metabolites in urine by liquid chromatography-tandem mass spectrometry with chiral derivatization. Anal Bioanal Chem 404:2427–2435. https://doi.org/10.1007/s00216-012-6385-9
Newmeyer MN, Concheiro M, Huestis MA (2014) Rapid quantitative chiral amphetamines liquid chromatography-tandem mass spectrometry: method in plasma and oral fluid with a cost-effective chiral derivatizing reagent. J Chromatogr A 1358:68–74. https://doi.org/10.1016/j.chroma.2014.06.096
Schwelm HM, Grumann C, Auwärter V, Neukamm M (2020) Application of a chiral HPLC-MS/MS method for the determination of 13 related amphetamine-type stimulants to forensic samples: Interpretative hypotheses. Drug Test Anal Accepted a: https://doi.org/10.1017/CBO9781107415324.004
J. Miller K, Gal J, Ames MM, (1984) High-performance liquid chromatographic resolution of enantiomers of 1-phenyl-2-aminopropanes (amphetamines) with four chiral reagents. J Chromatogr B Biomed Sci Appl 307:335–342. https://doi.org/10.1016/S0378-4347(00)84104-3
Sardela VF, Martucci MEP, de Araújo ALD, Leal EC, Oliveira DS, Carneiro GRA, Deventer K, Van Eenoo P, Pereira HMG, Aquino Neto FR (2018) Comprehensive analysis by liquid chromatography-Q-Orbitrap mass spectrometry: fast screening of peptides and organic molecules. J Mass Spectrom 53:476–503. https://doi.org/10.1002/jms.4077
World Anti-Doping Agency (2019a) Minimum required performance levels for detection and identification of non-threshold substances. WADA, Montreal
Sardela VF, Sardela PDO, Deventer K, Araújo ALD, Cavalcante KM, Padilha MC, Pereira HMG, Van Eenoo P, Aquino Neto FR (2013) Identification of sympathomimetic alkylamine agents in urine using liquid chromatography-mass spectrometry and comparison of derivatization methods for confirmation analyses by gas chromatography-mass spectrometry. J Chromatogr A 1298:76–85. https://doi.org/10.1016/j.chroma.2013.05.016
World Anti-Doping Agency (2019b) International Standard for Laboratories (ISL). WADA, Montreal
World Anti-Doping Agency (2015) Minimum criteria for chromatographic mass-spectrometric confirmation of the identity of analytes for doping control purposes. WADA, Montreal
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We are grateful to Bruna Cheble for the English review.
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This work was supported by CAPES.
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de Souza Anselmo, C., de Vasconcellos Silveira, C.M., Sardela, V.F. et al. Chiral Analysis of Amphetamine and Methamphetamine in Urine by Liquid Chromatography-Tandem Mass Spectrometry Applying Mosher Derivatization. Chromatographia 84, 47–52 (2021). https://doi.org/10.1007/s10337-020-03984-y
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DOI: https://doi.org/10.1007/s10337-020-03984-y