The present paper describes an original method for the online preconcentration and analysis of ring-substituted amphetamines in urine samples, used on the integration of robot-assisted hollow fiber liquid-phase microextraction (HF-LPME), high-performance liquid chromatography (HPLC), and fluorescence detection (FLD). A lab-made autosampler, actuating a 100-μL syringe and equipped with a three-way solenoid microvalve, allowed the acceptor phase to flow through and be withdrawn from the lumen fiber, enabling the automated online transference of the enriched acceptor phase for chromatographic analysis, through a six-port switching valve. The developed online HF-LPME-LC/FLD method demonstrated high analytical throughput and confidence, facilitating the efficient extraction and determination of the target analytes, with minimal solvent consumption and sample manipulation, in a straightforward way. Sample cleanup, analyte uptake, and analysis were carried out in 14.5 min. Under optimal conditions, automated online HF-LPME showed excellent linearity, precision, and trueness, obtaining intraday RSDs between 2.9 and 9.2% (n = 6) and interday RSDs between 5.3 and 9.3% (n = 6). Enrichment factors (EFs) ranged between 14.2 and 15.7, extraction recoveries (ERs) ranged between 17.7 and 19.5%, and the limits of detection (S/N = 3) were 2.0, 3.0, and 3.0 μg L−1 for MDA, MDMA, and MDEA, respectively. The method proved to be an effortless, rapid, reliable, and environment-friendly approach for the determination of drug abuse in urine samples.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
UNODC (United Nations Office on Drugs and Crime). World Drug Report 2018 Analysis of drug markets. World Drug Report 2018. 2018. 1–72 p.
Majchrzak M, Celiński R, Kuś P, Kowalska T, Sajewicz M. The newest cathinone derivatives as designer drugs: an analytical and toxicological review. Forensic Toxicol. 2018;36(1):33–50.
Taghvimi A, Dastmalchi S, Javadzadeh Y. Novel ceramic carbon-coated magnetic nanoparticles as stir bar sorptive extraction coating for simultaneous extraction of amphetamines from urine samples. Arab J Sci Eng. 2019;44(7):6373–80.
Bijlsma L, Celma A, López FJ, Hernández F. Monitoring new psychoactive substances use through wastewater analysis: current situation, challenges and limitations. Curr Opin Environ Sci Heal. 2019;9:1–12.
Celma A, Sancho JV, Salgueiro-González N, Castiglioni S, Zuccato E, Hernández F, Bijlsmaa L. Simultaneous determination of new psychoactive substances and illicit drugs in sewage: potential of micro-liquid chromatography tandem mass spectrometry in wastewater-based epidemiology. J Chromatogr A. 2019;1602:300–9.
Nemeškalová A, Bursová M, Sýkora D, Kuchař M, Čabala R, Hložek T. Salting out assisted liquid-liquid extraction for liquid chromatography tandem-mass spectrometry determination of amphetamine-like stimulants in meconium. J Pharm Biomed Anal. 2019;172:42–9.
Shin Y, Kong TY, Cheong JC, Kim JY, Lee JI, Lee HS. Simultaneous determination of 75 abuse drugs including amphetamines, benzodiazepines, cocaine, opioids, piperazines, zolpidem, and metabolites in human hair samples using liquid chromatography–tandem mass spectrometry. Biomed Chromatogr. 2019;33:e4600. https://doi.org/10.1002/bmc.4600.
Busardò FP, Gottardi M, Pacifici R, Varì MR, Tini A, Volpe AR, Giorgetti R, Pichini S. Nails Analysis for drugs used in the context of chemsex: a pilot study. J Anal Toxicol. 2019;1–6. https://doi.org/10.1093/jat/bkz009.
Akhoundian M, Alizadeh T, Ganjali MR, Norouzi P. Ultra-trace detection of methamphetamine in biological samples using FFT-square wave voltammetry and nano-sized imprinted polymer/MWCNTs -modified electrode. Talanta. 2019;200:115–23.
Song A, Yang J. Efficient determination of amphetamine and methylamphetamine in human urine using electro-enhanced single-drop microextraction with in-drop derivatization and gas chromatography. Anal Chim Acta. 2019;1045:162–8.
Ortuño J, Pizarro N, Farré M, Mas M, Segura J, Camí J, et al. Quantification of 3,4-methylenedioxymetamphetamine and its metabolites in plasma and urine by gas chromatography with nitrogen-phosphorus detection. J Chromatogr B Biomed Sci Appl. 1999;723(1–2):221–32.
Da Costa JL, Chasin AADM. Determination of MDMA, MDEA and MDA in urine by high performance liquid chromatography with fluorescence detection. J Chromatogr B Anal Technol Biomed Life Sci. 2004;811(1 SPEC. ISS):41–5.
Chalavi S, Asadi S, Nojavan S, Fakhari AR. Recent advances in microextraction procedures for determination of amphetamines in biological samples. Bioanalysis. 2019;11(5):437–60.
Malaca S, Rosado T, Restolho J, Rodilla JM, Rocha PMM, Silva L, et al. Determination of amphetamine-type stimulants in urine samples using microextraction by packed sorbent and gas chromatography-mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci. 2019;1120:41–50.
Sharifi V, Abbasi A, Nosrati A. Application of hollow fiber liquid phase microextraction and dispersive liquid-liquid microextraction techniques in analytical toxicology. J Food Drug Anal. 2016;24(2):264–76.
Darvish M, Qomi M, Akhgari M, Raoufi P. Determination of trace amounts of methamphetamine in biological samples by hollow fiber liquid-phase microextraction followed by high performance liquid chromatography. Biosci Biotechnol Res Asia. 2015;12(1):587–97.
Franco de Oliveira SCW de SE, Yonamine M. Measurement uncertainty for the determination of amphetamines in urine by liquid-phase microextraction and gas chromatography-mass spectrometry. Forensic Sci Int [Internet]. 2016;265:81–8. https://doi.org/10.1016/j.forsciint.2016.01.012.
Venson R, Korb AS, Cooper G. A review of the application of hollow-fiber liquid-phase microextraction in bioanalytical methods – a systematic approach with focus on forensic toxicology. J Chromatogr B Anal Technol Biomed Life Sci. 2019;1108(January):32–53.
Pedersen-Bjergaard SRK. Liquid-liquid microextraction for sample preparation of biological fluids prior to capillary electrophoresis. AnalChem. 1999;71(14):2650–6.
Gjelstad A. Three-phase hollow fiber liquid-phase microextraction and parallel artificial liquid membrane extraction. TrAC - Trends Anal Chem. 2019;113:25–31.
Esrafili A, Baharfar M, Tajik M, Yamini Y, Ghambarian M. Two-phase hollow fiber liquid-phase microextraction. TrAC - Trends Anal Chem. 2018;108:314–22.
Alexovič M, Horstkotte B, Solich P, Sabo J. Automation of static and dynamic non-dispersive liquid phase microextraction. Part 2: Approaches based on impregnated membranes and porous supports. Anal Chim Acta. 2016;907:18–30.
Esrafili A, Yamini Y, Ghambarian M, Ebrahimpour B. Automated preconcentration and analysis of organic compounds by on-line hollow fiber liquid-phase microextraction-high performance liquid chromatography. J Chromatogr A [Internet]. 2012;1262:27–33. https://doi.org/10.1016/j.chroma.2012.09.003.
Nazaripour A, Yamini Y, Ebrahimpour B, Fasihi J. Automated hollow-fiber liquid-phase microextraction followed by liquid chromatography with mass spectrometry for the determination of benzodiazepine drugs in biological samples. J Sep Sci. 2016;39(13):2595–603.
Ouyang G, Zhao W, Pawliszyn J. Automation and optimization of liquid-phase microextraction by gas chromatography. J Chromatogr A. 2007;1138(1–2):47–54.
Ouyang G, Pawliszyn J. Kinetic calibration for automated hollow fiber-protected liquid-phase microextraction. Anal Chem. 2006;78(16):5783–8.
Lopes D, Dias AN, Merib J, Carasek E. Hollow-fiber renewal liquid membrane extraction coupled with 96-well plate system as innovative high-throughput configuration for the determination of endocrine disrupting compounds by high-performance liquid chromatography-fluorescence and diode array de. Anal Chim Acta. 2018;1040:33–40.
Medina DAV, Rodríguez Cabal LF, Titato GM, Lanças FM, Santos-Neto ÁJ. Automated online coupling of robot-assisted single drop microextraction and liquid chromatography. J Chromatogr A. 2019;1595:66–72.
Medina DAV, Cabal LFR, Lanças FM, Santos-Neto ÁJ. Sample treatment platform for automated integration of microextraction techniques and liquid chromatography analysis. HardwareX [Internet]. 2019;6:1–16 Available from: https://www.sciencedirect.com/science/article/pii/S2468067218300907#b0010.
Jeannot MA, Cantwell FF. Solvent microextraction into a single drop. Anal Chem. 1996;68(13):2236–40.
Chao YY, Tu YM, Jian ZX, Wang HW, Huang YL. Direct determination of chlorophenols in water samples through ultrasound-assisted hollow fiber liquid-liquid-liquid microextraction on-line coupled with high-performance liquid chromatography. J Chromatogr A [Internet]. 2013;1271(1):41–9. https://doi.org/10.1016/j.chroma.2012.11.039.
Tang YQ, Weng N. Salting-out assisted liquid-liquid extraction for bioanalysis. Bioanalysis. 2013;5(12):1583–98.
Majors RE. Salting-out liquid-liquid extraction (SALLE). 2009
The authors received financial support from São Paulo Research Foundation (FAPESP) through grant nos. 2010/19910-9, 2014/03795-0, 2016/21950-5, 2017/02147-0, and 2018/00432-1, and from CNPq through grant nos. 459326/2014-7, 307293/2014-9, 311300/2015-4, and 425814/2018-1. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) – Finance Code 001.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the University of Campinas committee, CAAE 58187816.6.0000.5404, and with the 1964 Helsinki Declaration. Urine samples were obtained from healthy volunteers who signed informed consent form.
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
About this article
Cite this article
Cabal, L.F.R., Medina, D.A.V., Costa, J.L. et al. Determination of ring-substituted amphetamines through automated online hollow fiber liquid-phase microextraction-liquid chromatography. Anal Bioanal Chem 411, 7889–7897 (2019). https://doi.org/10.1007/s00216-019-02196-0
- Sample preparation
- Liquid chromatography
- Mass spectrometry