Skip to main content

Advertisement

SpringerLink
Log in

Development of a fully automated toxicological LC-MSn screening system in urine using online extraction with turbulent flow chromatography

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

In clinical toxicology, fast and specific methods are necessary for the screening of different classes of drugs. Therefore, an online extraction high-performance liquid chromatography coupled to mass spectrometry (LC-MSn) screening method using a MS2 and MS3 spectral library for the identification of xenobiotic substances has been developed and validated. Samples were run twice, once native and once after enzymatic hydrolysis. Internal standards and buffer were added to the urine samples. Following centrifugation, the supernatant was injected into the system. Extraction was performed by online turbulent flow chromatography. The chromatographic separation was achieved using a Phenyl/Hexyl column. For detection, a linear ion trap, equipped with an APCI interface, was used and the different compounds were identified using a MS2 and MS3 spectral library containing 356 compounds. The turnaround time to report the results of the screening including hydrolysis was approximately 2 h. About 92% of the 356 substances could be identified with a limit of identification below 100 ng/ml. The recovery and matrix effect experiments showed suitable results, and in six drug-free urine samples of healthy volunteers analyzed for selectivity, no substances have been identified. Carryover could be well controlled, and the method had a good reproducibility. The comparison of the results of 103 real patient urine samples showed a good agreement between the existing GC-MS and LC-MS methods with offline extraction and the new online extraction LC-MSn screening method. The presented method allows a fast and sensitive analysis of a broad range of compounds.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Maurer HH (2004) Position of chromatographic techniques in screening for detection of drugs or poisons in clinical and forensic toxicology and/or doping control. Clin Chem Lab Med 42:1310–1324

    Article  CAS  Google Scholar 

  2. Baden LR, Horowitz G, Jacoby H, Eliopoulos GM (2001) Quinolones and false-positive urine screening for opiates by immunoassay technology. JAMA 286:3115–3119

    Article  CAS  Google Scholar 

  3. Aebi B, Bernhard W (2002) Advances in the use of mass spectral libraries for forensic toxicology. J Anal Toxicol 26:149–156

    CAS  Google Scholar 

  4. Pragst F, Herzler M, Erxleben B (2004) Systematic toxicological analysis by high-performance liquid chromatography with diode array detection (HPLC-DAD). Clin Chem Lab Med 42:1325–1340

    Article  CAS  Google Scholar 

  5. Schönberg L, Grobosch T, Lampe D, Kloft C (2006) New screening method for basic compounds in urine by on-line extraction–high-performance liquid chromatography with photodiode-array detection. J Chromatogr A 1134:177–185

    Article  Google Scholar 

  6. Binder S, Regalia M, Biaggi McEachern M, Mazhar M (1989) Automated liquid chromatographic analysis of drugs in urine by on-line sample cleanup and isocratic muticolumn separation. J Chromatogr 473:325–341

    Article  CAS  Google Scholar 

  7. Kratzsch C, Tenberken O, Peters FT, Weber AA, Kraemer T, Maurer HH (2004) Screening, library-assisted identification and validated quantification of 23 benzodiazepines, flumazenil, zaleplone, zolpidem and zopiclone in plasma by liquid chromatography/mass spectrometry with atmospheric pressure chemical ionization. J Mass Spectrom 39:856–872

    Article  CAS  Google Scholar 

  8. Lips AG, Lameijer W, Fokkens RH, Nibbering NM (2001) Methodology for the development of a drug library based upon collision-induced fragmentation for the identification of toxicologically relevant drugs in plasma samples. J Chromatogr B Biomed Sci Appl 759:191–207

    Article  CAS  Google Scholar 

  9. Liu H-C, Liu RH, Lin D-L, Ho H-O (2010) Rapid screening and confirmation of drugs and toxic compounds in biological specimens using liquid chromatography/ion trap tandem mass spectrometry and automated library search. Rapid Commun Mass Spectrum 24:75–84

    Article  Google Scholar 

  10. Dulaurent S, Moesch C, Marquet P, Gaulier JM, Lachatre G (2010) Screening of pesticides in blood with liquid chromatography-linear ion trap mass spectrometry. Anal Bioanal Chem 396:2235–2249

    Article  CAS  Google Scholar 

  11. Sturm S, Hammann F, Drewe J, Maurer HH, Scholer A (2010) An automated screening method for drugs and toxic compounds in human serum and urine using liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 878:2726–2732

    Article  CAS  Google Scholar 

  12. Marquet P, Saint-Marcoux F, Gamble T, Leblanc J (2003) Comparison of a preliminary procedure for the general unknown screening of drugs and toxic compounds using a quadrupole-linear ion-trap mass spectrometer with a liquid chromatography-mass spectrometry reference technique. J Chromatogr B 789:9–18

    Article  CAS  Google Scholar 

  13. Mueller CA, Weinmann W, Dresen S, Schreiber A, Gergov M (2005) Development of a multi-target screening analysis for 301 drugs using a QTrap liquid chromatography/tandem mass spectrometry system and automated library searching. Rapid Commun Mass Spectrom 19:1332–1338

    Article  CAS  Google Scholar 

  14. Dresen S, Ferreiros N, Gnann H, Zimmermann R, Weinmann W (2010) Detection and identification of 700 drugs by multi-target screening with a 3200 Q TRAP LC-MS/MS system and library searching. Anal Bioanal Chem 396:2425–2434

    Article  CAS  Google Scholar 

  15. Lynch KL, Breaud AR, Vandenberghe H, Wu AH, Clarke W (2010) Performance evaluation of three liquid chromatography mass spectrometry methods for broad spectrum drug screening. Clin Chim Acta 411:1474–1481

    Article  CAS  Google Scholar 

  16. Dresen S, Gergov M, Politi L, Halter C, Weinmann W (2009) ESI-MS/MS library of 1, 253 compounds for application in forensic and clinical toxicology. Anal Bioanal Chem 395:2521–2526

    Article  CAS  Google Scholar 

  17. Jansen R, Lachatre G, Marquet P (2005) LC-MS/MS systematic toxicological analysis: comparison of MS/MS spectra obtained with different instruments and settings. Clin Biochem 38:362–372

    Article  CAS  Google Scholar 

  18. Hopley C, Bristow T, Lubben A, Simpson A, Bull E, Klagkou K, Herniman J, Langley J (2008) Towards a universal product ion mass spectral library - reproducibility of product ion spectra across eleven different mass spectrometers. Rapid Commun Mass Spectrom 22:1779–1786

    Article  CAS  Google Scholar 

  19. Oberacher H, Pavlic M, Libiseller K, Schubert B, Sulyok M, Schuhmacher R, Csaszar E, Kofeler HC (2009) On the inter-instrument and inter-laboratory transferability of a tandem mass spectral reference library: 1. Results of an Austrian multicenter study. J Mass Spectrom 44:485–493

    Article  CAS  Google Scholar 

  20. Oberacher H, Pavlic M, Libiseller K, Schubert B, Sulyok M, Schuhmacher R, Csaszar E, Kofeler HC (2009) On the inter-instrument and the inter-laboratory transferability of a tandem mass spectral reference library: 2. Optimization and characterization of the search algorithm. J Mass Spectrom 44:494–502

    Article  CAS  Google Scholar 

  21. Eichhorst JC, Etter ML, Rousseaux N, Lehotay DC (2009) Drugs of abuse testing by tandem mass spectrometry: a rapid, simple method to replace immunoassays. Clin Biochem 42:1531–1542

    Article  CAS  Google Scholar 

  22. Lee HK, Ho CS, Iu YPH, Lai PSJ, Shek CC, Lo Y-C, Klinke HB, Wood M (2009) Development of a broad toxicological screening technique for urine using ultra-performance liquid chromatography and time-of-flight mass spectrometry. Anal Chim Acta 649:80–90

    Article  CAS  Google Scholar 

  23. Ojanperä S, Pelander A, Pelzing M, Krebs I, Vuori E, Ojanperä I (2006) Isotopic pattern and accurate mass determination in urine drug screening by liquid chromatography/time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 20:1161–1167

    Article  Google Scholar 

  24. Decaestecker T, Vande Casteele S, Wallemacq P, Van Peteghem C, Defore D, Van Bocxlaer J (2004) Information-dependent acquisition-mediated LC-MS/MS screening procedure with semiquantitative potential. Anal Chem 76:6365–6373

    Article  CAS  Google Scholar 

  25. Ceglarek U, Lembcke J, Martin Fiedler G, Werner M, Witzigmann H, Peter Hauss J, Thiery J (2004) Rapid simultaneous quantification of immunosuppressants in transplant patients by turbulent flow chromatography combined with tandem mass spectrometry. Clin Chim Acta 346:181–190

    Article  CAS  Google Scholar 

  26. Breaud AR, Harlan R, Di Bussolo JM, McMillin GA, Clarke W (2010) A rapid and fully-automated method for the quantitation of tricyclic antidepressants in serum using turbulent-flow liquid chromatography-tandem mass spectrometry. Clin Chim Acta 411:825–832

    Article  CAS  Google Scholar 

  27. Dams R, Huestis MA, Lambert WE, Murphy CM (2003) Matrix effect in bio-analysis of illicit drugs with LC-MS/MS: influence of ionization type, sample preparation, and biofluid. J Am Soc Mass Spectrom 14:1290–1294

    Article  CAS  Google Scholar 

  28. Zgraggen S, Bonatim R, Gutteck-Amsler U, Rentsch KM (2007) Toxicological screening after the REMEDI™—comparison of a GC-MS screening with the REMEDI™. Ther Drug Monit 29:492–492

    Google Scholar 

  29. Mueller DM, Rentsch KM (2009) Development and validation of a library-assisted toxicological screening method in urine by LC-MS2. In: Proceedings XVI GTFCh-Symposium. pp. 166–169

  30. Bonfiglio R, King R, Olah T, Merkle K (1999) The effects of sample preparation methods on the variability of the electrospray ionization response for model drug compounds. Rapid Commun Mass Spectrom 13:1175–1185

    Article  CAS  Google Scholar 

  31. Matuszewski BK, Constanzer ML, Chavez-Eng CM (2003) Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal Chem 75:3019–3030

    Article  CAS  Google Scholar 

  32. Sauvage F-L (2006) Screening of drugs and toxic compounds with liquid chromatography-linear ion trap tandem mass spectrometry. Clin Chem 52:1735–1742

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Thermo Fisher Scientific for providing the LC-MS system and the online extraction instrument.

Author information

Authors and Affiliations

  1. Institute for Clinical Chemistry, University Hospital Zurich, Raemistrasse 100, 8091, Zurich, Switzerland

    Daniel M. Mueller & Katharina M. Rentsch

  2. Thermo Fisher Scientific, 91963, Les Ulis Cédex, France

    Bénédicte Duretz

  3. Thermo Fisher Scientifc, Franklin, MA, 02038, USA

    Francois A. Espourteille

Authors
  1. Daniel M. Mueller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bénédicte Duretz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Francois A. Espourteille
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Katharina M. Rentsch
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Katharina M. Rentsch.

Additional information

Published in the special issue Forensic Toxicology with Guest Editors Frank T. Peters, Hans H. Maurer, and Frank Musshoff.

Electronic supplementary materials

Below is the link to the electronic supplementary material.

ESM 1

(PDF 4,727 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mueller, D.M., Duretz, B., Espourteille, F.A. et al. Development of a fully automated toxicological LC-MSn screening system in urine using online extraction with turbulent flow chromatography. Anal Bioanal Chem 400, 89–100 (2011). https://doi.org/10.1007/s00216-010-4560-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-010-4560-4

Keywords

Search

Navigation