An integrated microfluidic device for solid-phase extraction and spectrophotometric detection of opium alkaloids in urine samples

  • Ali Farahani
  • Hassan SereshtiEmail author
Research Paper


A novel lab-on-chip integrated microfluidic device for solid-phase extraction (SPE) and spectrophotometric detection of morphine (MOR), codeine (COD), and papaverine (PAP) was developed. The extracted analytes were analyzed with a miniature UV-Vis spectrophotometer. The SPE adsorptive phase composed of polyurethane/polyaniline (PU/PANI) nanofibers was fabricated by electrospinning and in situ oxidative polymerization techniques. The sorbent was characterized by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). The main factors of extraction such as desorption conditions, pH, salt effect, and extraction time were investigated. The partial least square (PLS) regression was applied to improve the quantification of analytes. The linear dynamic ranges (LDRs) for MOR, COD, and PAP were 4–240, 4–210, and 1–150 ng mL–1, respectively. Finally, the proposed method was successfully applied for the determination of MOR, COD, and PAP in human urine samples and the extraction recoveries were obtained in the range of 66.7–85.0% with RSDs < 8.3%.


On-chip solid-phase extraction Opiates Microfluidics Point-of-care testing Polymer electrospun nanofiber 


Funding information

The authors would like to thank the Iran National Science Foundation (INSF) for the financial support through the Research Grants.

Compliance with ethical standards

The human urine sample was obtained from volunteers, with informed consent provided to the Shariati Hospital (Tehran, Iran). The studies have been performed in accordance with the ethical standards approved by the appropriate research ethics committee of University of Tehran.

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2019_2214_MOESM1_ESM.pdf (193 kb)
ESM 1 (PDF 192 kb)


  1. 1.
    Whitesides GM. The origins and the future of microfluidics. Nature. 2006;442:368–73.PubMedCrossRefGoogle Scholar
  2. 2.
    Sackmann EK, Fultonand AL, Beebe DJ. The present and future role of microfluidics in biomedical research. Nature. 2014;507:181–9.PubMedCrossRefGoogle Scholar
  3. 3.
    Pagaduan JV, Sahore V, Woolley AT. Applications of microfluidics and microchip electrophoresis for potential clinical biomarker analysis. Anal Bioanal Chem. 2015;407(23):6911–22.PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Nouri N, Khorram P, Sereshti H. Applications of three-dimensional graphenes for preconcentration, extraction, and sorption of chemical species: a review. Microchim Acta. 2019;186:232.CrossRefGoogle Scholar
  5. 5.
    Esfandiarnejad R, Sereshti H, Farahani A. Polyaniline immobilized on polycaprolactam nanofibers as a sorbent in electrochemically controlled solid-phase microextraction coupled with HPLC for the determination of angiotensin II receptor antagonists in human blood plasma. Anal Bioanal Chem. 2019;7:1-0.Google Scholar
  6. 6.
    Mauk M, Song J, Liu C, Bau H. Simple approaches to minimally-instrumented, microfluidic-based point-of-care nucleic acid amplification tests. Biosensors. 2018;8(1):1–30.7.CrossRefGoogle Scholar
  7. 7.
    Chen X, Cui DF, Zhang LL, Li H, Sun JH, Cai HY. A porous microfluidic chip for protein extraction based on solid phase extraction method. Key Eng Mater. 2011;483:297–300.CrossRefGoogle Scholar
  8. 8.
    Hu G, Lee JS, Li D. A microfluidic fluorous solid-phase extraction chip for purification of amino acids. J. Colloid Interface Sci. 2006;301(2):697–702.PubMedCrossRefPubMedCentralGoogle Scholar
  9. 9.
    Hwang KY, Kwon SH, Jung SO, Namkoong K, Jung WJ, Kim JH, et al. Solid phase DNA extraction with a flexible bead-packed microfluidic device to detect methicillin-resistant Staphylococcus aureus in nasal swabs. Anal. Chem. 2012;84(18):7912–8.PubMedCrossRefPubMedCentralGoogle Scholar
  10. 10.
    Sahore V, Sonker M, Nielsen AV, Knob R, Kumar S, Woolley AT. Automated microfluidic devices integrating solid-phase extraction, fluorescent labeling, and microchip electrophoresis for preterm birth biomarker analysis. Anal Bioanal Chem. 2018;410(3):933–41.PubMedCrossRefGoogle Scholar
  11. 11.
    Park M, Seo TS. An integrated microfluidic device with solid-phase extraction and graphene oxide quantum dot array for highly sensitive and multiplex detection of trace metal ions. Biosens Bioelectron. 2019;126:405–11.PubMedCrossRefGoogle Scholar
  12. 12.
    Choi K, Mudrik JM, Wheeler AR. A guiding light: spectroscopy on digital microfluidic devices using in-plane optical fibre waveguides. Anal Bioanal Chem. 2015;407(24):7467–75.PubMedCrossRefGoogle Scholar
  13. 13.
    Azzouz T, Tauler R. Application of multivariate curve resolution alternating least squares (MCR-ALS) to the quantitative analysis of pharmaceutical and agricultural samples. Talanta. 2008;74:1201–10.PubMedCrossRefGoogle Scholar
  14. 14.
    Lynch KB, Chen A, Liu S. Miniaturized high-performance liquid chromatography instrumentation. Talanta. 2018;177:94–103.PubMedCrossRefGoogle Scholar
  15. 15.
    Lu T, Yuan Y, He X, Li M, Pu X, Xu T, et al. Simultaneous determination of multiple spectrophotometry and a partial least squares. RSC Adv. 2015;5:13021–7.Google Scholar
  16. 16.
    Dinis-Oliveira RJ. Metabolism and metabolomics of opiates: a long way of forensic implications to unravel. J Forensic Leg Med. 2019;61:128–40.PubMedCrossRefGoogle Scholar
  17. 17.
    Yaksh TL. Spinal opiates: a review of their effect on spinal function with emphasis on pain processing. Acta Anaesthesiol Scand. 1987;31:25–37.CrossRefGoogle Scholar
  18. 18.
    Ricardo Buenaventura M, Rajive Adlaka M, Nalini SM. Opioid complications and side effects. Pain physician. 2008;11:S105–20.PubMedGoogle Scholar
  19. 19.
    Boland JW, Johnson M, Ferreira D, Berry DJ. In silico (computed) modelling of doses and dosing regimens associated with morphine levels above international legal driving limits. Palliat Med. 2018;32(7):1222–32.PubMedPubMedCentralCrossRefGoogle Scholar
  20. 20.
    Fabricant DS, Farnsworth NR. The value of plants used in traditional medicine for drug discovery. Environ Health Perspect. 2001;109:69–75.PubMedPubMedCentralGoogle Scholar
  21. 21.
    Sindrup SH, Brøsen K. The pharmacogenetics of codeine hypoalgesia. Pharmacogenetics. 1995;6:335–46.CrossRefGoogle Scholar
  22. 22.
    Aleksa K, Walasek P, Fulga N, Kapur B, Gareri J, Koren G. Simultaneous detection of seventeen drugs of abuse and metabolites in hair using solid phase micro extraction (SPME) with GC/MS. Forensic Sci Int. 2012;218:31–6.PubMedCrossRefPubMedCentralGoogle Scholar
  23. 23.
    Kohler I, Schappler J, Sierro T, Rudaz S. Dispersive liquid–liquid microextraction combined with capillary electrophoresis and time-of-flight mass spectrometry for urine analysis. J Pharm Biomed Anal. 2013;73:82–9.PubMedCrossRefPubMedCentralGoogle Scholar
  24. 24.
    Yamini Y, Pourali A, Seidi S, Rezazadeh M. Electromembrane extraction followed by high performance liquid chromatography: an efficient method for extraction and determination of morphine, oxymorphone, and methylmorphine from urine samples. Anal Methods. 2014;6(15):5554–65.CrossRefGoogle Scholar
  25. 25.
    Gholivand MB, Jalalvand AR, Goicoechea HC, Gargallo R, Skov T, Paimard G. Combination of electrochemistry with chemometrics to introduce an efficient analytical method for simultaneous quantification of five opium alkaloids in complex matrices. Talanta. 2015;131:26–37.PubMedCrossRefPubMedCentralGoogle Scholar
  26. 26.
    Baciu T, Borrull F, Aguilar C, Calull M. Findings in the hair of drug abusers using pressurized liquid extraction and solid-phase extraction coupled in-line with capillary electrophoresis. J Pharm Biomed Anal. 2016;131:420–8.PubMedCrossRefPubMedCentralGoogle Scholar
  27. 27.
    Mohseni N, Bahram M, Baheri T. Chemical nose for discrimination of opioids based on unmodified gold nanoparticles. Sens Actuators B. 2017;250:509–17.CrossRefGoogle Scholar
  28. 28.
    Rosa C, Elizabeth H, Enrique L, Luis CJ, Juana C, Antonio RJ, et al. Optomicrofluidic system for spectrophotometric analysis: automated process and wireless. In: 2018 IEEE 2nd Colombian Conference on Robotics and Automation (CCRA); 2018. p. 1–6.Google Scholar
  29. 29.
    Zainal Alam MNH, Jaya Kumar J, John Whyte D, Doeven EH, Kouzani A. A portable sensor for cell optical density measurement in microfluidic chips. Meas Control. 2018;51(7-8):213–22.CrossRefGoogle Scholar
  30. 30.
    Ding Z, Zhang D, Wang G, Tang M, Dong Y, Zhang Y, et al. An in-line spectrophotometer on a centrifugal microfluidic platform for real-time protein determination and calibration. Lab Chip. 2016;16(18):3604–14.PubMedCrossRefGoogle Scholar
  31. 31.
    Fronczek CF, San Park T, Harshman DK, Nicolini AM, Yoon JY. Paper microfluidic extraction and direct smartphone-based identification of pathogenic nucleic acids from field and clinical samples. RSC Adv. 2014;4(22):11103–10.CrossRefGoogle Scholar
  32. 32.
    Songjaroen T, Maturos T, Sappat A, Tuantranont A, Laiwattanapaisal W. Portable microfluidic system for determination of urinary creatinine. Anal Chim Acta. 2009;647(1):78–83.PubMedCrossRefGoogle Scholar
  33. 33.
    Tian M, Wang Y, Qu L, Zhu S, Han G, Zhang X, et al. Electromechanical deformation sensors based on polyurethane/polyaniline electrospinning nanofibrous mats. Synth Met. 2016;219:11–9.CrossRefGoogle Scholar
  34. 34.
    Eskandarpour N, Sereshti H. Electrospun polyurethane fibers doped with manganese oxide nanoparticles as an effective adsorbent for determination of priority pollutant mono-nitrophenols in water samples. J Environ Chem Eng. 2019;7(1):102926.CrossRefGoogle Scholar
  35. 35.
    Hong KH, Oh KW, Kang TJ. Preparation of conducting nylon-6 electrospun fiber webs by the in situ polymerization of polyaniline. J Appl Polym Sci. 2005;96:983–91.CrossRefGoogle Scholar
  36. 36.
    Nouri N, Sereshti H, Farahani A. Graphene-coated magnetic-sheet solid-phase extraction followed by high-performance liquid chromatography with fluorescence detection for the determination of aflatoxins B1, B2, G1, and G2 in soy-based samples. J Sep Sci. 2018;41(16):3258–66.PubMedCrossRefGoogle Scholar
  37. 37.
    Fan Q, Zhang X, Qin Z. Preparation of polyaniline/polyurethane fibers and their piezoresistive property. J. Macromol. Sci. Part B: Phys. 2012;51(4):736–46.Google Scholar
  38. 38.
    Butoi B, Groza A, Dinca P, Balan A, Barna V. Morphological and structural analysis of polyaniline and poly (o-anisidine) layers generated in a DC glow discharge plasma by using an oblique angle electrode deposition configuration. Polymers. 2017;9(12):732.PubMedCentralCrossRefPubMedGoogle Scholar
  39. 39.
    Wang LL, Lin YW, Wang XF, Xiao N, Xu YD, Li HD, et al. A selective review and comparison for interval variable selection in spectroscopic modeling. Chemom. Intell. Lab. Syst. 2018;15(172):229–40.CrossRefGoogle Scholar
  40. 40.
    Lin X, Wang J, Li L, Wang X, Lü H, Xie Z. Separation and determination of five major opium alkaloids with mixed mode of hydrophilic/cation-exchange monolith by pressurized capillary electrochromatography. J Sep Sci. 2007;30(17):3011–7.PubMedCrossRefPubMedCentralGoogle Scholar
  41. 41.
    Menardo C, Nechtschein M, Roussean A, Travers JP, Hany P. Investigation on the structure of polyaniline: 13C nmr and titration studies. Synth Met. 1988;25:311–22.CrossRefGoogle Scholar
  42. 42.
    Kang XJ, Chen LQ, Zhang YY, Liu YW, Gu ZZ. Performance of electrospun nanofibers for SPE of drugs from aqueous solutions. J Sep Sci. 2008;31(18):3272–8.PubMedCrossRefPubMedCentralGoogle Scholar
  43. 43.
    Soltani MD, Taher MA, Behzadi M, Fazelirad H. Synthesis, characterization and application of magnetic carbon nanotubes for the simultaneous extraction and high performance liquid chromatographic determination of codeine and morphine in human urine, blood serum, opium and tablet samples. Sens Actuators A. 2018;280:31–7.CrossRefGoogle Scholar
  44. 44.
    Barroso M, Dias M, Vieira DN, López-Rivadulla M, Queiroz JA. Simultaneous quantitation of morphine, 6-acetylmorphine, codeine, 6-acetylcodeine and tramadol in hair using mixed-mode solid-phase extraction and gas chromatography–mass spectrometry. Anal Bioanal Chem. 2010;396(8):3059–69.PubMedCrossRefPubMedCentralGoogle Scholar
  45. 45.
    Taylor K, Elliott S. A validated hybrid quadrupole linear ion-trap LC–MS method for the analysis of morphine and morphine glucuronides applied to opiate deaths. Forensic Sci Int. 2009;187:34–41.PubMedCrossRefPubMedCentralGoogle Scholar
  46. 46.
    Dams R, Benijts T, Lambert WE, De Leenheer AP. Simultaneous determination of in total 17 opium alkaloids and opioids in blood and urine by fast liquid chromatography–diode-array detection–fluorescence detection, after solid-phase extraction. J Chromatogr B: Anal Technol Biomed Life Sci. 2002;773:53–61.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Chemistry, College of ScienceUniversity of TehranTehranIran

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