Analytical and Bioanalytical Chemistry

, Volume 410, Issue 20, pp 5071–5083 | Cite as

Fully automated sample preparation procedure to measure drugs of abuse in plasma by liquid chromatography tandem mass spectrometry

  • Tiphaine Robin
  • Alan Barnes
  • Sylvain Dulaurent
  • Neil Loftus
  • Sigrid Baumgarten
  • Stéphane Moreau
  • Pierre Marquet
  • Souleiman El Balkhi
  • Franck Saint-MarcouxEmail author
Research Paper


For the analysis of drugs and pharmaceutical compounds in biological matrices, extraction procedures are typically used for LC-MS/MS analysis often requiring manual steps in sample preparation. In this study, we report a fully automated extraction method carried out by a programable liquid handler directly coupled to an LC-MS/MS system for the determination of 42 components (illicit drugs and/or metabolites) (plus 20 deuterated internal standards). The acquisition was performed in positive ionization mode with up to 15 MRM transitions per compound, each with optimized collision energy (MRM spectrum mode) to enable qualitative library searching in addition to quantitation. After placing the sample tube into the system, no further intervention was necessary: automated preparation used 50 μL of blood or plasma with 3 μL of extracted sample injected for analysis. The method was validated according to the requirements of ISO 15189. The limit of detection and quantification was 1–5 ng/mL depending on the compound. Stability experiments found that historic calibration curve data files could accurately quantify for up to 1 month with less than 20% uncertainty. Comparison to a QuEChERS method was made using patient samples providing a regression correlation R2 = 0.98 between the two methods. This approach was successfully designed to support parallel sample preparation and analysis therefore significantly increasing sample throughput and reduced cycle times.

Graphical abstract


Liquid chromatography Mass spectrometry Automated sample preparation Drugs of abuse 



Drugs of abuse


Internal standard


Lower limit of detection


Lower limit of quantitation


Multiple reaction monitoring


Upper limit of quantitation


Retention time


Compliance with ethical standards

The study has been carried out using biological samples of the biobank “CRBioLim” of the CHU of Limoges, authorized by regulatory authorities (French Ministry of Research and Regional Health Agency) and ethical committee “CPP SOOM 4” with the reference AC 2016-2748, according to the French regulation. All the samples stored in “CRBioLim” have been given by patients who did not oppose to the use of these samples for the development of an analytical method, as allowed by the French regulation (mainly the law on bioethics n° 2004-800 of August 6, 2004).

Conflict of interest

Author, Tiphaine Robin, is funded 50% by CIFRE and 50% by Shimadzu Corporation. All authors declare that they have no conflict of interest.


  1. 1.
    Cailleux A, Le Bouil A, Auger B, Bonsergent G, Turcant A, Allain P. Determination of opiates and cocaine and its metabolites in biological fluids by high-performance liquid chromatography with electrospray tandem mass spectrometry. J Anal Toxicol. 1999;23:620–4.CrossRefPubMedGoogle Scholar
  2. 2.
    Moeller MR, Steinmeyer S, Kraemer T. Determination of drugs of abuse in blood. J Chromatogr B Biomed Sci Appl. 1998;713:91–109.CrossRefPubMedGoogle Scholar
  3. 3.
    Soriano T, Jurado C, Menendez M, Repetto M. Improved solid phase extraction method for systematic toxicological analysis in biological fluids. J Anal Toxicol. 2001;25:137–43.CrossRefPubMedGoogle Scholar
  4. 4.
    De Jager AD, Bailey NL. Online extraction LC–MS/MS method for the simultaneous quantitative confirmation of urine drugs of abuse and metabolites: amphetamines, opiates, cocaine, cannabis, benzodiazepines and methadone. J Chromatogr B. 2011;879:2642–52.CrossRefGoogle Scholar
  5. 5.
    Saussereau E, Lacroix C, Gaulier JM, Goulle JP. On-line liquid chromatography/tandem mass spectrometry simultaneous determination of opiates, cocainics and amphetamines in dried blood spots. J Chromatogr B. 2012;885–886:17.Google Scholar
  6. 6.
    Imbert L, Dulaurent S, Mercerolle M, Morichon J, Lachâtre G, Gaulier JM. Development and validation of a single LC-MS/MS assay following SPE for simultaneous hair analysis of amphetamines, opiates, cocaine and metabolites. Forensic Sci Int. 2014;234:132–8.CrossRefPubMedGoogle Scholar
  7. 7.
    Anzillotti L, Odoardi S, Strano-Rossi S. Cleaning up blood samples using a modified “QuEChERS” procedure for the determination of drugs of abuse and benzodiazepines by UPLC–MSMS. Forensic Sci Int. 2014;243:99–106.CrossRefPubMedGoogle Scholar
  8. 8.
    Dulaurent S, El Balkhi S, Poncelet L, Gaulier JM, Marquet P, Saint-Marcoux F. QuEChERS sample preparation prior to LC-MS/MS determination of opiates, amphetamines, and cocaine metabolites in whole blood. Anal Bioanal Chem. 2016;408:1467–74.CrossRefPubMedGoogle Scholar
  9. 9.
    Wissenbach DK, Meyer MR, Remane D, Philipp AA, Weber AA, Maurer HH. Drug of abuse screening in urine as part of metabolite-based LC-MS screening concept. Anal Bional Chem. 2011;400:89–100.CrossRefGoogle Scholar
  10. 10.
    Bourgogne E, Grivet C, Varesio E, Hopfgartner G. Generic on-line solid phase extraction sample preparation strategies for the analysis of drugs in biological matrices by LC-MS/MS. J Pharm Biomed Anal. 2015 Jan;102:290–8.CrossRefPubMedGoogle Scholar
  11. 11.
    Vogeser M, Kirchhoff F. Progress in automation of LC-MS in laboratory medicine. Clin Biochem. 2011;44:4–13.CrossRefPubMedGoogle Scholar
  12. 12.
    Bjork, et al. Quantification of 31 illicit and medicinal drugs and metabolites in whole blood by fully automated solid-phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry. Anal Bional Chem. 2013;405:2607–17.CrossRefGoogle Scholar
  13. 13.
    Poncelet L, El Bakhi S, Dulaurent S, Saint-Marcoux F. QuEChERS sample preparation prior to LC–MS/MS determination of benzodiazepines. Toxicol Anal Clin. 2016;28:201–10.Google Scholar
  14. 14.
    Sauvage FL, Gaulier JM, Lachatre G, Marquet P. Pitfalls and prevention strategies for liquid chromatography–tandem mass spectrometry in the selected reaction–monitoring mode for drug analysis. Clin Chem. 2008;54:1519–27.CrossRefPubMedGoogle Scholar
  15. 15.
    Remane D, Wissenbach DK, Peters FT. Recent advances of liquid chromatography-(tandem) mass spectrometry in clinical and forensic toxicology—an update. Clin Biochem. 2016;49:1051–71.CrossRefPubMedGoogle Scholar
  16. 16.
    Marquet P, Venisse M, Lacassie E, Lachatre G. In-source ID mass spectral libraries for the “general unknown” screening of drugs and toxicants. Analysis. 2000;28:925–34.Google Scholar
  17. 17.
    Anastassiades M, Lehotay SJ, Stajnbaher D, Schenck FJ. Fast and easy multiresidue method employing acetonitrile extraction/partitioning and “dispersive solid-phase extraction” for the determination of pesticide residues in produce. J AOAC Int. 2003;86:412–31.PubMedGoogle Scholar
  18. 18.
    Matsuta S, Nakanishi K, Miki A, Zaitsu K, Shima N, Kamata T, et al. Development of a simple one-pot extraction method for various drugs and metabolites of forensic interest in blood by modifying the QuEChERS method. Forensic Sci Int. 2013;232:40–5.CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Tiphaine Robin
    • 1
  • Alan Barnes
    • 2
  • Sylvain Dulaurent
    • 1
  • Neil Loftus
    • 2
  • Sigrid Baumgarten
    • 3
  • Stéphane Moreau
    • 3
  • Pierre Marquet
    • 1
  • Souleiman El Balkhi
    • 1
  • Franck Saint-Marcoux
    • 1
    Email author
  1. 1.Department of Pharmacology and ToxicologyLimoges University HospitalLimogesFrance
  2. 2.Shimadzu CorporationManchesterUK
  3. 3.Shimadzu Europa GmbHDuisburgGermany

Personalised recommendations