Skip to main content

Volumetric absorptive microsampling as an alternative tool for therapeutic drug monitoring of first-generation anti-epileptic drugs

A Correction to this article was published on 15 February 2018

This article has been updated

Abstract

Dosage adjustment of anti-epileptic drugs by therapeutic drug monitoring (TDM) is very useful, especially for the first-generation anti-epileptic drugs (AEDs). Microsampling—the collection of small volumes of blood—is increasingly considered a valuable alternative to conventional venous sampling for TDM. Volumetric absorptive microsampling (VAMS) allows accurate and precise collection of a fixed volume of blood, eliminating the volumetric blood hematocrit bias coupled to conventional dried blood spot collection. The aim of this study was to develop and validate an LC-MS/MS method for the determination and quantification of four anti-epileptic drugs (carbamazepine, valproic acid, phenobarbital, and phenytoin) and one active metabolite (carbamazepine-10,11-epoxide) in samples collected by VAMS. The method was fully validated based on international guidelines. Precision (%RSD) was below 10%, while, with a single exception, accuracy (%bias) met the acceptance criteria. Neither carry-over nor unacceptable interferences were observed, the method being able to distinguish between the isomers oxcarbazepine and carbamazepine-10,11-epoxide. All compounds were stable in VAMS samples for at least 1 month when stored at room temperature, 4 °C, and − 20 °C and for at least 1 week when stored at 60 °C. Internal standard-corrected matrix effects were below 10%, with %RSDs below 4%. High (> 85%) recovery values were obtained and the effect of the hematocrit on the recovery was overall limited. Successful application on external quality control materials and on left-over patient samples demonstrated the validity and applicability of the developed procedure.

Graphical representation of the sampling, chemical structures, and the resulting chromatogram for volumetric absorptive microsampling (VAMS)-based therapeutic drug monitoring of first-generation anti-epileptic drugs by liquid chromatography with tandem mass spectrometric detection.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Change history

  • 15 February 2018

    We would like to call the reader’s attention to the fact that unfortunately in fig. 2 of the original article the figure headings of both graphs are the same.

References

  1. 1.

    Milosheska D, Grabnar I, Vovk T. Dried blood spots for monitoring and individualization of antiepileptic drug treatment. Eur J Pharm Sci. 2015;75:25–39.

    CAS  Article  Google Scholar 

  2. 2.

    Krasowski MD, McMillin GA. Advances in anti-epileptic drug testing. Clin Chim Acta. 2014;436:224–36.

    CAS  Article  Google Scholar 

  3. 3.

    Velghe S, Capiau S, Stove CP. Opening the toolbox of alternative sampling strategies in clinical routine: a key-role for (LC-)MS/MS. Trac Trend Anal Chem. 2016;84:61–73.

    CAS  Article  Google Scholar 

  4. 4.

    Capiau S, Alffenaar J-W, Stove CP. Alternative sampling strategies for therapeutic drug monitoring. In: Clarke W, Dasgupta A, editors. Clinical challenges in therapeutic drug monitoring. Amsterdam: Elsevier; 2016. p. 279–336.

    Chapter  Google Scholar 

  5. 5.

    Shah NM, Hawwa AF, Millership JS, Collier PS, McElnay JC. A simple bioanalytical method for the quantification of antiepileptic drugs in dried blood spots. J Chromatogr B Analyt Technol Biomed Life Sci. 2013;923-924:65–73.

    CAS  Article  Google Scholar 

  6. 6.

    Popov TV, Maricic LC, Prosen H, Voncina DB. Development and validation of dried blood spots technique for quantitative determination of topiramate using liquid chromatography-tandem mass spectrometry. Biomed Chromatogr. 2013;27(8):1054–61.

    Google Scholar 

  7. 7.

    Linder C, Andersson M, Wide K, Beck O, Pohanka A. A LC-MS/MS method for therapeutic drug monitoring of carbamazepine, lamotrigine and valproic acid in DBS. Bioanalysis. 2015;7(16):2031–9.

    CAS  Article  Google Scholar 

  8. 8.

    Villanelli F, Giocaliere E, Malvagia S, Rosati A, Forni G, Funghini S, et al. Dried blood spot assay for the quantification of phenytoin using liquid chromatography-mass spectrometry. Clin Chim Acta. 2015;440:31–5.

    CAS  Article  Google Scholar 

  9. 9.

    Shokry E, Villanelli F, Malvagia S, Rosati A, Forni G, Funghini S, et al. Therapeutic drug monitoring of carbamazepine and its metabolite in children from dried blood spots using liquid chromatography and tandem mass spectrometry. J Pharm Biomed Anal. 2015;109:164–70.

    CAS  Article  Google Scholar 

  10. 10.

    AbuRuz S, Al-Ghazawi M, Al-Hiari Y. A simple dried blood spot assay for therapeutic drug monitoring of lamotrigine. Chromatographia. 2010;71(11–12):1093–9.

    CAS  Article  Google Scholar 

  11. 11.

    la Marca G, Malvagia S, Filippi L, Luceri F, Moneti G, Guerrini R. A new rapid micromethod for the assay of phenobarbital from dried blood spots by LC-tandem mass spectrometry. Epilepsia. 2009;50(12):2658–62.

    Article  Google Scholar 

  12. 12.

    la Marca G, Malvagia S, Filippi L, Fiorini P, Innocenti M, Luceri F, et al. Rapid assay of topiramate in dried blood spots by a new liquid chromatography-tandem mass spectrometric method. J Pharm Biomed Anal. 2008;48(5):1392–6.

    Article  Google Scholar 

  13. 13.

    Wilhelm AJ, den Burger JC, Swart EL. Therapeutic drug monitoring by dried blood spot: progress to date and future directions. Clin Pharmacokinet. 2014;53(11):961–73.

    CAS  Article  Google Scholar 

  14. 14.

    Fan L, Lee JA. Managing the effect of hematocrit on DBS analysis in a regulated environment. Bioanalysis. 2012;4(4):345–7.

    CAS  Article  Google Scholar 

  15. 15.

    De Kesel PM, Capiau S, Lambert WE, Stove CP. Current strategies for coping with the hematocrit problem in dried blood spot analysis. Bioanalysis. 2014;6(14):1871–4.

    Article  Google Scholar 

  16. 16.

    Youhnovski N, Bergeron A, Furtado M, Garofolo F. Pre-cut dried blood spot (PCDBS): an alternative to dried blood spot (DBS) technique to overcome hematocrit impact. Rapid Commun Mass Spectrom. 2011;25(19):2951–8.

    CAS  Article  Google Scholar 

  17. 17.

    De Kesel PM, Capiau S, Stove VV, Lambert WE, Stove CP. Potassium-based algorithm allows correction for the hematocrit bias in quantitative analysis of caffeine and its major metabolite in dried blood spots. Anal Bioanal Chem. 2014;406(26):6749–55.

    Article  Google Scholar 

  18. 18.

    den Burger JC, Wilhelm AJ, Chahbouni AC, Vos RM, Sinjewel A, Swart EL. Haematocrit corrected analysis of creatinine in dried blood spots through potassium measurement. Anal Bioanal Chem. 2015;407(2):621–7.

    Article  Google Scholar 

  19. 19.

    Li Y, Henion J, Abbott R, Wang P. The use of a membrane filtration device to form dried plasma spots for the quantitative determination of guanfacine in whole blood. Rapid Commun Mass Spectrom. 2012;26(10):1208–12.

    CAS  Article  Google Scholar 

  20. 20.

    Li F, Zulkoski J, Fast D, Michael S. Perforated dried blood spots: a novel format for accurate microsampling. Bioanalysis. 2011;3(20):2321–33.

    CAS  Article  Google Scholar 

  21. 21.

    Meesters RJ, Zhang J, van Huizen NA, Hooff GP, Gruters RA, Luider TM. Dried matrix on paper disks: the next generation DBS microsampling technique for managing the hematocrit effect in DBS analysis. Bioanalysis. 2012;4(16):2027–35.

    CAS  Article  Google Scholar 

  22. 22.

    Leuthold LA, Heudi O, Deglon J, Raccuglia M, Augsburger M, Picard F, et al. New microfluidic-based sampling procedure for overcoming the hematocrit problem associated with dried blood spot analysis. Anal Chem. 2015;87(4):2068–71.

    CAS  Article  Google Scholar 

  23. 23.

    De Kesel PM, Lambert WE, Stove CP. Does volumetric absorptive microsampling eliminate the hematocrit bias for caffeine and paraxanthine in dried blood samples? A comparative study. Anal Chim Acta. 2015;881:65–73.

    Article  Google Scholar 

  24. 24.

    Denniff P, Spooner N. Volumetric absorptive microsampling: a dried sample collection technique for quantitative bioanalysis. Anal Chem. 2014;86(16):8489–95.

    CAS  Article  Google Scholar 

  25. 25.

    Verougstraete N, Lapauw B, Van Aken S, Delanghe J, Stove C, Stove V. Volumetric absorptive microsampling at home as an alternative tool for the monitoring of HbA1c in diabetes patients. Clin Chem Lab Med. 2017;55(3):462–9.

    CAS  Article  Google Scholar 

  26. 26.

    Kok MGM, Fillet M. Volumetric absorptive microsampling: current advances and applications. J Pharm Biomed Anal. 2018;147:288–96.

  27. 27.

    Atugonza R, Kakooza-Mwesige A, Lhatoo S, Kaddumukasa M, Mugenyi L, Sajatovic M, et al. Multiple anti-epileptic drug use in children with epilepsy in Mulago hospital, Uganda: a cross sectional study. BMC Pediatr. 2016;16:34.

    Article  Google Scholar 

  28. 28.

    Gogtay NJ, Kshirsagar NA, Dalvi SS. Therapeutic drug monitoring in a developing country: an overview. Br J Clin Pharmacol. 2001;52(Suppl 1):103S–8S.

    Article  Google Scholar 

  29. 29.

    Bertilsson L. Clinical pharmacokinetics of carbamazepine. Clin Pharmacokinet. 1978;3(2):128–43.

    CAS  Article  Google Scholar 

  30. 30.

    U.S. Department of Health and Human Services. Food and Drug Administration. Center for Drug Evaluation and Research. Center for Veterinary Medicine. Draft Guidance for Industry. Bioanalytical Method Validation. 2013. https://www.fda.gov/downloads/Drugs/Guidances/ucm368107.pdf. Accessed June 2017.

  31. 31.

    European Medicines Agency. Guideline on Bioanalytical Method Validation. 2015. http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/08/WC500109686.pdf. Accessed June 2017.

  32. 32.

    Wille SMR, Peters FT, Di Fazio V, Samyn N. Practical aspects concerning validation and quality control for forensic and clinical bioanalytical quantitative methods. Accred Qual Assur. 2011;16(6):279–92.

  33. 33.

    Abu-Rabie P, Denniff P, Spooner N, Chowdhry BZ, Pullen FS. Investigation of different approaches to incorporating internal standard in DBS quantitative bioanalytical workflows and their effect on nullifying hematocrit-based assay bias. Anal Chem. 2015;87(9):4996–5003.

    CAS  Article  Google Scholar 

  34. 34.

    Houts T. Immunochromatography. In: Price CP, editor. Principles and practice of immunoassay. New York: Stockton Press; 1991. p. 563–83.

    Chapter  Google Scholar 

  35. 35.

    Launiainen T, Ojanpera I. Drug concentrations in post-mortem femoral blood compared with therapeutic concentrations in plasma. Drug Test Anal. 2014;6(4):308–16.

    CAS  Article  Google Scholar 

  36. 36.

    Morris RG, Schapel GJ. Phenytoin and phenobarbital assayed by the ACCULEVEL method compared with EMIT in an outpatient clinic setting. Ther Drug Monit. 1988;10(4):469–73.

    CAS  Article  Google Scholar 

  37. 37.

    Linder C, Wide K, Walander M, Beck O, Gustafsson LL, Pohanka A. Comparison between dried blood spot and plasma sampling for therapeutic drug monitoring of antiepileptic drugs in children with epilepsy: a step towards home sampling. Clin Biochem. 2017;50(7–8):418–24.

    CAS  Article  Google Scholar 

  38. 38.

    Kong ST, Lim SH, Lee WB, Kumar PK, Wang HY, Ng YL, et al. Clinical validation and implications of dried blood spot sampling of carbamazepine, valproic acid and phenytoin in patients with epilepsy. PLoS One. 2014;9(9):e108190.

    Article  Google Scholar 

  39. 39.

    Rhoden L, Antunes MV, Hidalgo P, Alvares da Silva C, Linden R. Simple procedure for determination of valproic acid in dried blood spots by gas chromatography-mass spectrometry. J Pharm Biomed Anal. 2014;96:207–12.

    CAS  Article  Google Scholar 

Download references

Acknowledgements

The authors wish to acknowledge Prof. Veronique Stove, PharmD. Matthijs Oyaert and their team for assistance with blood collection and all volunteers who participated in the study. Furthermore, SV would also like to thank the Special Research Fund (BOF) for granting her a PhD fellowship (application number 01D42414).

Author information

Affiliations

Authors

Corresponding author

Correspondence to Christophe P. Stove.

Ethics declarations

Approval for this study was provided by the Ethics Committee of Ghent University Hospital (EC2017/0572). For the use of left-over samples to evaluate an alternative procedure for AED monitoring, the need to obtained individual informed consent was waived by the Ethics Committee.

Conflict of interest

The author declares to not have any financial, commercial, legal, or professional relationship with other organizations, or with the people working with them, that could influence the matter discussed in this manuscript.

Additional information

The original version of this article was revised: the headings of figure 2 of both graphs are the same.

A correction to this article is available online at https://doi.org/10.1007/s00216-018-0951-8.

Electronic supplementary material

ESM 1

(PDF 345 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Velghe, S., Stove, C.P. Volumetric absorptive microsampling as an alternative tool for therapeutic drug monitoring of first-generation anti-epileptic drugs. Anal Bioanal Chem 410, 2331–2341 (2018). https://doi.org/10.1007/s00216-018-0866-4

Download citation

Keywords

  • Anti-epileptic drugs
  • Volumetric absorptive microsampling
  • Liquid chromatography-tandem mass spectrometry
  • Sample preparation
  • Alternative sampling strategies