Skip to main content
SpringerLink
Log in

Simultaneous determination of 20 drugs of abuse in oral fluid using ultrasound-assisted dispersive liquid–liquid microextraction

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

Abstract

Drugs of abuse and new psychoactive substances (NPS) for recreational purposes are in constant evolution, and their consumption constitutes a significant risk to public health and road safety. The development of an analytical methodology to confirm the intake of illicit drugs in biological fluids is required for an effective control of these substances. An ultra-performance liquid chromatography–tandem mass spectrometry method (UPLC-MS/MS) was developed for simultaneous determination of 10 synthetic cathinones and 10 illicit drugs in oral fluid easily sampled through non-invasive maneuvers. The UPLC-MS/MS method was coupled to an ultrasound-assisted dispersive liquid–liquid microextraction (US-DLLME), which is a miniaturized and inexpensive technique that uses reduced volumes of solvents and samples. The US-DLLME was optimized by using a 213441//18 asymmetric screening design and a Doehlert design. Sample volume, dispersion and extraction solvent volumes, pH, US time, and amount of sodium chloride were evaluated. The US-DLLME-UPLC-MS/MS method was validated according to international guidelines. Limits of quantitation (LOQs) ranged from 0.25 to 5 ng mL−1, and the linear range spanned from LOQ to 500 ng mL−1 with R2 higher than 0.9907, for most of the target drugs. Precision ranged from 1.7 to 14.8 %RSD. Accuracy, i.e., extraction recovery, ranged from 74 to 129%. The proposed method was successfully applied to the analysis of 15 samples from patients on a drug detoxification program.

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
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. European Monitoring Centre for Drugs and Drug Addiction, European Drug Report 2016: trends and developments, Publications Office of the European Union, Luxembourg, 2016 http://www.emcdda.europa.eu/system/files/publications/2637/TDAT16001ENN.pdf

  2. Miller BL, Stogner JM, Miller JM, Fernandez MI. The arrest and synthetic novel psychoactive drug relationship: observations from a young adult population. J Drug Issues. 2017;47:91–103. https://doi.org/10.1177/0022042616678611.

    Article  Google Scholar 

  3. Rech MA, Donahey E, Cappiello Dziedzic JM, Oh L, Greenhalgh E. New drugs of abuse. Pharmacotherapy. 2015;35:189–97. https://doi.org/10.1002/phar.1522.

    Article  CAS  PubMed  Google Scholar 

  4. Benítez G, March-Salas M, Villa-Kamel A, Cháves-Jiménez U, Hernández J, Montes-Osuna N, et al. The genus Datura L. (Solanaceae) in Mexico and Spain—ethnobotanical perspective at the interface of medical and illicit uses. J Ethnopharmacol. 2018;219:133–51. https://doi.org/10.1016/j.jep.2018.03.007.

    Article  PubMed  Google Scholar 

  5. Bosker WM, Huestis MA. Oral fluid testing for drugs of abuse. Clinical Chem. 2009;55:1910–31. https://doi.org/10.1373/clinchem.2008.108670.

    Article  CAS  Google Scholar 

  6. Aldlgan AA, Torrance HJ. Bioanalytical methods for the determination of synthetic cannabinoids and metabolites in biological specimens. TRAC Trends Anal Chem. 2016;80:444–57. https://doi.org/10.1016/j.trac.2016.03.025.

    Article  CAS  Google Scholar 

  7. Mercolini L, Protti M. Biosampling strategies for emerging drugs of abuse: towards the future of toxicological and forensic analysis. J Pharm Biomed Anal. 2016;130:202–19. https://doi.org/10.1016/j.jpba.2016.06.046.

    Article  CAS  PubMed  Google Scholar 

  8. Amaratunga P, Lemberg BL, Lemberg D. Quantitative measurement of synthetic cathinones in oral fluid. J Anal Toxicol. 2013;37:622–8. https://doi.org/10.1093/jat/bkt080.

    Article  CAS  PubMed  Google Scholar 

  9. Di Rago M, Chu M, Rodda LN, Jenkins E, Kotsos A, Gerostamoulos D. Ultra-rapid targeted analysis of 40 drugs of abuse in oral fluid by LC-MS/MS using carbon-13 isotopes of methamphetamine and MDMA to reduce detector saturation. Anal Bioanal Chem. 2016;40:3737–49. https://doi.org/10.1007/s00216-016-9458-3.

    Article  CAS  Google Scholar 

  10. Sergi M, Montesano C, Odoardi S, Rocca LM, Fabrizi G, Compagnone D, et al. Microextraction by packed sorbent coupled to liquid chromatography tandem mass spectrometry for the rapid and sensitive determination of cannabinoids in oral fluids. J Chromatogr A. 2013;1301:139–46. https://doi.org/10.1016/j.chroma.2013.05.072.

    Article  CAS  PubMed  Google Scholar 

  11. Newmeyer MN, Concheiro M, Huestis MA. Rapid quantitative chiral amphetamines liquid chromatography-tandem mass spectrometry: method in plasma and oral fluid with a cost-effective chiral derivatizing reagent. J Chromatogr A. 2014;1358:68–74. https://doi.org/10.1016/j.chroma.2014.06.096.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Montesano C, Simeoni MC, Curini R, Sergi M, Lo Sterzo C, Compagnone D. Determination of illicit drugs and metabolites in oral fluid by microextraction on packed sorbent coupled with LC-MS/MS. Anal Bioanal Chem. 2015;407:3647–58. https://doi.org/10.1007/s00216-015-8583-8.

    Article  CAS  PubMed  Google Scholar 

  13. Liu HC, Lee HT, Hsu YC, Huang MH, Liu RH, Chen TJ, et al. Direct injection LC-MS-MS analysis of opiates, methamphetamine, buprenorphine, methadone and their metabolites in oral fluid from substitution therapy patients. J Anal Toxicol. 2015;39:472–80. https://doi.org/10.1093/jat/bkv041.

    Article  CAS  PubMed  Google Scholar 

  14. de Castro A, Lendoiro E, Fernández-Vega H, Steinmeyer S, López-Rivadulla M, Cruz A. Liquid chromatography tandem mass spectrometry determination of selected synthetic cathinones and two piperazines in oral fluid. Cross reactivity study with an on-site immunoassay device. J Chromatogr A. 2014;1374:93–101. https://doi.org/10.1016/j.chroma.2014.11.024.

    Article  CAS  PubMed  Google Scholar 

  15. Mercolini L, Protti M, Catapano MC, Rudge J, Sberna AE. LC-MS/MS and volumetric absorptive microsampling for quantitative bioanalysis of cathinone analogs in dried urine, plasma and oral fluid samples. J Pharm Biomed Anal. 2016;123:186–94. https://doi.org/10.1016/j.jpba.2016.02.015.

    Article  CAS  PubMed  Google Scholar 

  16. Rezaee M, Assadi Y, MilaniHosseini MR, Aghaee E, Ahmadi F, Berijani S. Determination of organic compounds in water using dispersive liquid-liquid microextraction. J Chromatogr A, 2006;1116:1–9. https://doi.org/10.1016/j.chroma.2006.03.007

  17. Jain R, Singh R. Applications of dispersive liquid-liquid micro-extraction in forensic toxicology. TRAC Trends Anal Chem. 2016;75:227–37. https://doi.org/10.1016/j.trac.2015.07.007.

    Article  CAS  Google Scholar 

  18. Fisichella M, Odoardi S, Strano-Rossi S. High-throughput dispersive liquid/liquid microextraction (DLLME) method for the rapid determination of drugs of abuse, benzodiazepines and other psychotropic medications in blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and application to forensic cases. Microchem J. 2015;123:33–41. https://doi.org/10.1016/j.microc.2015.05.009.

    Article  CAS  Google Scholar 

  19. World Medical Association Declaration of Helsinki ethical principles for medical research involving human subjects. JAMA J Am Med Assoc. 2013;310:2191–4. https://doi.org/10.1001/jama.2013.281053.

  20. Mathieu D, Nony J, Phan-Tan-Luu R. NemrodW®, Version 2011, LPRAI, Marseille

  21. Lewis GA, Mathieu D, Phan-Tan-Luu R. Pharmaceutical experimental design. New York: Marcel Dekker; 1999. https://doi.org/10.1016/S0378-5173(99)00384-1.

    Book  Google Scholar 

  22. U.S. Department of Health and Human Services Food and Drug Administration, Guidance for industry: bioanalytical method validation, draft guidance, U.S. Department of Health and Human Services Food and Drug Administration, 2013. https://www.fda.gov/downloads/Drugs/%20GuidanceComplianceRegulatoryInformation/Guidances/UCM368107.pdf (Accessed on June 2018).

  23. U.S. Food and Drug Administration Foods and Veterinary Medicine Science and Research Steering Committee, Guidelines for the validation of chemical methods, 2015. https://www.fda.gov/downloads/scienceresearch/fieldscience/ucm273418.pdf (Accessed on June 2018).

  24. ICH harmonized tripartite guideline: validation of analytical procedures: text and methodology Q2(R1), International Conference of Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use, 2005. https://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm265700.htm (Accessed on June 2018).

  25. Magnusson B, Örnemark U. Eurachem guide: the fitness for purpose of analytical methods—a laboratory guide to method validation and related topics. 2nd ed. Teddington: Eurachem; 2014.

    Google Scholar 

  26. Kruve A, Rebane R, Kipper K, Oldekop M-L, Evard H, Herodes K, et al. Tutorial review on validation of liquid chromatography–mass spectrometry methods: part II. Anal Chim Acta. 2015;870:8–28. https://doi.org/10.1016/j.aca.2015.02.016.

    Article  CAS  PubMed  Google Scholar 

  27. Mansour FR, Khairy MA. Pharmaceutical and biomedical applications of dispersive liquid-liquid microextraction. J Chromatogr B. 2017;1061–1062:382–91. https://doi.org/10.1016/j.jchromb.2017.07.055.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Professor M.I. Loza (BioFarma Research Group–USC) for assistance with the instrumentation.

Funding

This work was funded by Ministerio del Interior-DGT (Project SPIP2015-01838) of Spain and the European Regional Development Fund (FEDER).

Author information

Authors and Affiliations

  1. Forensic Toxicology Service, Faculty of Medicine, Institute of Legal Medicine, University of Santiago de Compostela, 15782, Santiago de Compostela, Spain

    P. Fernández & M. Regenjo

  2. Department of Analytical Chemistry, Faculty of Chemistry and Health Research Institute of Santiago de Compostela (IDIS), University of Santiago de Compostela, 15782, Santiago de Compostela, Spain

    A. Ares, R. A. Lorenzo & A. M. Carro

  3. Drug Addiction Assistance Unit, Monforte de Lemos, 27400, Lugo, Spain

    A. M. Fernández

Authors
  1. P. Fernández
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Regenjo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Ares
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. M. Fernández
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R. A. Lorenzo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. M. Carro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to R. A. Lorenzo or A. M. Carro.

Ethics declarations

This study has been carried out using oral fluid samples from healthy volunteers and patients participating in substitution therapy programs, authorized by the Comité Autonómico de Ética da Investigación de Galicia Consellería de Sanidade. Xunta de Galicia (SPAIN), with the reference 2015/488, according to Spanish Regulation Law 14/2007, of biomedical investigation and the Real Decreto 1716/2011, of November 18. All the samples have been provided by volunteers and patients who did not oppose to the use of these samples for the development of an analytical method and signed the corresponding informed consent.

Conflict of interest

All authors declare that they have no conflict of interest.

Electronic supplementary material

ESM 1

(PDF 973 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fernández, P., Regenjo, M., Ares, A. et al. Simultaneous determination of 20 drugs of abuse in oral fluid using ultrasound-assisted dispersive liquid–liquid microextraction. Anal Bioanal Chem 411, 193–203 (2019). https://doi.org/10.1007/s00216-018-1428-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-018-1428-5

Keywords

Search

Navigation