Analytical and Bioanalytical Chemistry

, Volume 402, Issue 9, pp 2805–2815 | Cite as

Microfluidic chip based nano liquid chromatography coupled to tandem mass spectrometry for the determination of abused drugs and metabolites in human hair

  • Kevin Y. Zhu
  • K. Wing Leung
  • Annie K. L. Ting
  • Zack C. F. Wong
  • Winki Y. Y. Ng
  • Roy C. Y. Choi
  • Tina T. X. Dong
  • Tiejie Wang
  • David T. W. Lau
  • Karl W. K. TsimEmail author
Original Paper


A microfluidic chip based nano-HPLC coupled to tandem mass spectrometry (nano-HPLC-Chip-MS/MS) has been developed for simultaneous measurement of abused drugs and metabolites: cocaine, benzoylecgonine, cocaethylene, norcocaine, morphine, codeine, 6-acetylmorphine, phencyclidine, amphetamine, methamphetamine, MDMA, MDA, MDEA, and methadone in the hair of drug abusers. The microfluidic chip was fabricated by laminating polyimide films and it integrated an enrichment column, an analytical column and a nanospray tip. Drugs were extracted from hairs by sonication, and the chromatographic separation was achieved in 15 min. The drug identification and quantification criteria were fulfilled by the triple quardropule tandem mass spectrometry. The linear regression analysis was calibrated by deuterated internal standards with all of the R 2 at least over 0.993. The limit of detection (LOD) and the limit of quantification (LOQ) were from 0.1 to 0.75 and 0.2 to 1.25 pg/mg, respectively. The validation parameters including selectivity, accuracy, precision, stability, and matrix effect were also evaluated here. In conclusion, the developed sample preparation method coupled with the nano-HPLC-Chip-MS/MS method was able to reveal the presence of drugs in hairs from the drug abusers, with the enhanced sensitivity, compared with the conventional HPLC-MS/MS.


Microfluidic chip LC-MS Drug abuse Hair analysis Method development 



This research was supported by grants from the Office of the Vice-President for Research and Graduate Studies (VPRD011SC01), Croucher Foundation (CAS-CF07/08.SC03), Beat Drug Fund (BDF101014), and ITC (GHP/018/10SZ) awarded to KT.

Supplementary material

216_2012_5711_MOESM1_ESM.pdf (327 kb)
ESM 1 (PDF 327 kb)


  1. 1.
    Terry SC, Jerman JH, Angell JB (1979) A gas chromatographic air analyzer fabricated on a silicon wafer. IEEE Trans Electron Devices 26:1880–1886CrossRefGoogle Scholar
  2. 2.
    Manz A, Miyahara Y, Miura J, Watanabe Y, Miyagi H, Sato K (1990) Design of an open-tubular column liquid chromatograph using silicon chip technology. Sens Actuators, B Chem 1:249–255CrossRefGoogle Scholar
  3. 3.
    Ericson C, Holm J, Ericson T, Hjerté S (2000) Electroosmosis- and pressure-driven chromatography in chips using continuous beds. Anal Chem 72:81–87CrossRefGoogle Scholar
  4. 4.
    Lazar IM, Trisiripisal P, Sarvaiya HA (2006) Microfluidic liquid chromatography system for proteomic applications and biomarker screening. Anal Chem 78:5513–5524CrossRefGoogle Scholar
  5. 5.
    Fuentes HV, Woolley AT (2007) Electrically actuated, pressure-driven liquid chromatography separations in microfabricated devices. Lab Chip Miniaturisation Chem Biol 7:1524–1531CrossRefGoogle Scholar
  6. 6.
    Hasselbrink EF Jr, Shepodd TJ, Rehm JE (2002) High-pressure microfluidic control in lab-on-a-chip devices using mobile polymer monoliths. Anal Chem 74:4913–4918CrossRefGoogle Scholar
  7. 7.
    Reichmuth DS, Shepodd TJ, Kirby BJ (2004) On-chip high-pressure picoliter injector for pressure-driven flow through porous media. Anal Chem 76:5063–5068CrossRefGoogle Scholar
  8. 8.
    Fortier MH, Bonneil E, Goodley P, Thibault P (2005) Integrated microfluidic device for mass spectrometry-based proteomics and its application to biomarker discovery programs. Anal Chem 77:1631–1640CrossRefGoogle Scholar
  9. 9.
    Reichmuth DS, Shepodd TJ, Kirby BJ (2005) Microchip HPLC of peptides and proteins. Anal Chem 77:2997–3000CrossRefGoogle Scholar
  10. 10.
    Giusti P, Lobinski R, Szpunar J, Schaumlöffel D (2006) Development of a nebulizer for a sheathless interfacing of NanoHPLC and ICPMS. Anal Chem 78:965–971CrossRefGoogle Scholar
  11. 11.
    Ehlert S, Kraiczek K, Mora JA, Dittmann M, Rozing GP, Tallarek U (2008) Separation efficiency of particle-packed HPLC microchips. Anal Chem 80:5945–5950CrossRefGoogle Scholar
  12. 12.
    Hardouin J, Duchateau M, Joubert-Caron R, Caron M (2006) Usefulness of an integrated microfluidic device (HPLC-Chip-MS) to enhance confidence in protein identification by proteomics. Rapid Commun Mass Spectrom 20:3236–3244CrossRefGoogle Scholar
  13. 13.
    Groleau PE, Desharnais P, Coté L, Ayotte C (2008) Low LC-MS/MS detection of glycopeptides released from pmol levels of recombinant erythropoietin using nanoflow HPLC-chip electrospray ionization. J Mass Spectrom 43:924–935CrossRefGoogle Scholar
  14. 14.
    Flamini R, De Rosso M, Smaniotto A, Panighel A, Vedova AD, Seraglia R, Traldi P (2009) Fast analysis of isobaric grape anthocyanins by chip-liquid chromatography/mass spectrometry. Rapid Commun Mass Spectrom 23:2891–2896CrossRefGoogle Scholar
  15. 15.
    Bai HY, Lin SL, Chan SA, Fuh MR (2010) Characterization and evaluation of two-dimensional microfluidic chip-HPLC coupled to tandem mass spectrometry for quantitative analysis of 7-aminoflunitrazepam in human urine. Analyst 135:2737–2742CrossRefGoogle Scholar
  16. 16.
    Bush DM (2008) The U.S. Mandatory Guidelines for Federal Workplace Drug Testing Programs: current status and future considerations. Forensic Sci Int 174:111–119CrossRefGoogle Scholar
  17. 17.
    Department of Health and Human Services (2004) Mandatory guidelines for federal workplace drug-testing programs, Federal Register, 69, pp 19675Google Scholar
  18. 18.
    Yegles M, Wennig R (2007) In: Kintz P (ed) Analytical and practical aspects of drug testing in hair. Taylor & Francis Group, Boca RatonGoogle Scholar
  19. 19.
    Skender L, Karačić V, Brčić I, Bagarić A (2002) Quantitative determination of amphetamines, cocaine, and opiates in human hair by gas chromatography/mass spectrometry. Forensic Sci Int 125:120–126CrossRefGoogle Scholar
  20. 20.
    Cingolani M, Scavella S, Mencarelli R, Mirtella D, Froldi R, Rodriguez D (2004) Simultaneous detection and quantitation of morphine, 6-acetylmorphine, and cocaine in toenails: comparison with hair analysis. J Anal Toxicol 28:128–131Google Scholar
  21. 21.
    Scheidweiler KB, Huestis MA (2004) Simultaneous quantification of opiates, cocaine, and metabolites in hair by LC-APCI-MS/MS. Anal Chem 76:4358–4363CrossRefGoogle Scholar
  22. 22.
    Yazdi AS, Es’haghi Z (2005) Surfactant enhanced liquid-phase microextraction of basic drugs of abuse in hair combined with high performance liquid chromatography. J Chromatogr A 1094:1–8CrossRefGoogle Scholar
  23. 23.
    Pujadas M, Pichini S, Poudevida S, Menoyo E, Zuccaro P, Farré M, De La Torre R (2003) Development and validation of a gas chromatography-mass spectrometry assay for hair analysis of amphetamine, methamphetamine and methylenedioxy derivatives. J Chromatogr B Anal Technol Biomed Life Sci 798:249–255CrossRefGoogle Scholar
  24. 24.
    Stanaszek R, Piekoszewski W (2004) Simultaneous determination of eight underivatized amphetamines in hair by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (HPLC-APCI-MS). J Anal Toxicol 28:77–85Google Scholar
  25. 25.
    US Food and Drug Administration (2001) Guidance for industry: bioanalytical method validation. New Hampshire Ave Silver Spring. Accessed 5 May 2001
  26. 26.
    Frank TP (2006) In: Aldo P (ed) Applications of LC–MS in toxicology, Pharmaceutical, LondonGoogle Scholar
  27. 27.
    Wu YH, Lin KI, Chen SC, Chang YZ (2008) Integration of GC/EI-MS and GC/NCI-MS for simultaneous quantitative determination of opiates, amphetamines, MDMA, ketamine, and metabolites in human hair. J Chromatogr B Anal Technol Biomed Life Sci 870:192–202CrossRefGoogle Scholar
  28. 28.
    Kronstrand R, Nyström I, Strandberg J, Druid H (2004) Screening for drugs of abuse in hair with ion spray LC-MS-MS. Forensic Sci Int 145:183–190CrossRefGoogle Scholar
  29. 29.
    European Workplace Drug Testing Society (2010) Drug and alcohol testing in hair, collection and analysis. Validated from August 2010

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Kevin Y. Zhu
    • 1
  • K. Wing Leung
    • 1
  • Annie K. L. Ting
    • 1
  • Zack C. F. Wong
    • 1
  • Winki Y. Y. Ng
    • 1
  • Roy C. Y. Choi
    • 1
  • Tina T. X. Dong
    • 1
  • Tiejie Wang
    • 1
    • 2
  • David T. W. Lau
    • 1
  • Karl W. K. Tsim
    • 1
    Email author
  1. 1.Division of Life Science, and Center for Chinese MedicineThe Hong Kong University of Science and TechnologyClear Water Bay RoadHong Kong
  2. 2.Shenzhen Institute for Drug ControlShenzhenChina

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