Original Article


How to cite: Müller, Lizot LLF, Linden R. Profile of post-graduates in legal and forensic from medical colleges in Sao Paulo. Persp Med Legal Pericia Med. 2022; 7: e221223


Submitted 11/05/2022
Accepted 12/20/2022

The authors inform no conflict of interest.


Isadora Ritter Müller (1)


Lilian de Lima Feltraco Lizot (2)

http://lattes.cnpq.br/2523163224304535  – https://orcid.org/0000-0003-3165-1407

Rafael Linden (3)


(1) Universidade Feevale, Laboratório de Toxicologia e Análises Toxicológicas, Novo Hamburgo-RS, Brasil. (autor principal)

(2) Universidade Feevale, Laboratório de Toxicologia e Análises Toxicológicas, Novo Hamburgo-RS, Brasil.(tratamento de dados)

(3) Universidade Feevale, Laboratório de Toxicologia e Análises Toxicológicas, Novo Hamburgo-RS, Brasil.(orientador)

Email: isa_muller@hotmail.com


Introduction: Individuals undergoing treatment for epilepsy may have difficulty adhering to treatment. Among the reasons, forgetfulness and adverse effects caused by medication can be highlighted. With this scenario, it is understood that there is a need to carry out therapeutic monitoring of patients undergoing treatment for epilepsy. The OneDraw device, which allows you to obtain a capillary blood sample from its fixation in the upper part of the arm, eliminates some of the obstacles encountered in collecting capillary blood by the finger, such as pain, discomfort and waiting time. Material and methods: The validation of a method in High Performance Liquid Chromatography coupled to Mass Spectrometry (HPLC-MS/MS) was carried out, following the validation parameters of the Food and Drug Administration for the quantification of carbamazepine (CBZ) and Valproic acid (AV) in dry blood spots. Result: The method demonstrated high sensitivity for the detection of CBZ and AV in capillary blood samples. The detection limit of the method was 1.0 mg L-1 for CBZ and 10 mg L-1 for AV. Intra-assay precision was 3.29 to 6.75% for CBZ and 4.67 to 6.54% for AV. Discussion: Although other techniques exist, the use of HPLC-MS/MS is the main technique used for the analysis of anticonvulsants in DBS due to its high sensitivity and selectivity. Conclusion: Therefore, a simple and sensitive method is being validated for CBZ and AV quantification in capillary blood samples collected with the OneDraw device.

Keywords: Anticonvulsivants, Dried Blood Spots, Liquid Cromatography.

Bibliographical references

  1. WHO. Epilepsy [Internet]. 2019. p. 1–6. Available at: https://www.who.int/news-room/fact-sheets/detail/epilepsy
  2. Neels HM, Sierens AC, Naelaerts K, Scharpé SL, Hatfield GM, Lambert WE. Therapeutic drug monitoring of old and newer anti-epileptic drugs. Clin Chem Lab Med [Internet]. 2004;42(11):1228–55. Available at: 10.1515/CCLM.2004.245
  3. Marino Jr. R, Cukiert A, Pinho E. Aspectos epidemiológicos da epilepsia em São Paulo: um estudo da prevalência. Arq Neuropsiquiatr [Internet]. 1986;44(3):243–54. Available at: https://doi.org/10.1590/S0004-282X1986000300004
  4. Costa OL, Brandão EC, Segundo BML. Atualização em epilepsia: revisão de literatura. Rev Med. 2020;99(2):170–81. http://dx.doi.org/10.11606/issn.1679-9836.v99i2p170-181
  5. Noronha ALA, Borges MA, Marques LHN, Zanetta DMT, Fernandes PT, De Boer H, et al. Prevalence and pattern of epilepsy treatment in different socioeconomic classes in Brazil. Epilepsia. 2007;48(5):880–5. https://doi.org/10.1111/j.1528-1167.2006.00974.x
  6. Thijs RD, Surges R, O’Brien TJ, Sander JW. Epilepsy in adults. Lancet (London, England). fevereiro de 2019;393(10172):689–701. https://doi.org/10.1016/S0140-6736(18)32596-0
  7. Moshé SL, Perucca E, Ryvlin P, Tomson T. Epilepsy: new advances. Lancet (London, England). março de 2015;385(9971):884–98. https://doi.org/10.1016/S0140-6736(14)60456-6
  8. Beydoun A, Dupont S, Zhou D, Matta M, Nagire V, Lagae L. Seizure : European Journal of Epilepsy Current role of carbamazepine and oxcarbazepine in the management of epilepsy. Seizure Eur J Epilepsy [Internet]. 2020;83(December):251–63. Available at: https://doi.org/10.1016/j.seizure.2020.10.018
  9. Sankaraneni R, Lachhwani D. Antiepileptic drugs-a review. Pediatr Ann. 2015;44(2):e36–42. https://doi.org/10.3928/00904481-20150203-10
  10. Ding J, Zhang Y, Jiao Z, Wang Y. The effect of poor compliance on the pharmacokinetics of carbamazepine and its epoxide metabolite using Monte Carlo simulation. Acta Pharmacol Sin. novembro de 2012;33(11):1431–40. https://doi.org/10.1038/aps.2012.135
  11. Ghodke-Puranik Y, Thorn CF, Lamba JK, Leeder JS, Song W, Birnbaum AK, et al. Valproic acid pathway: Pharmacokinetics and pharmacodynamics. Pharmacogenet Genomics. 2013;23(4):236–41. doi: 10.1097/FPC.0b013e32835ea0b2
  12. Patsalos PN, Spencer EP, Berry DJ. Therapeutic drug monitoring of antiepileptic drugs in epilepsy: A 2018 update. Vol. 40, Therapeutic Drug Monitoring. 2018. 526–548 p. doi: 10.1097/FTD.0000000000000546
  13. Landmark CJ, Johannessen SI, Patsalos PN. Expert Opinion on Drug Metabolism & Toxicology Therapeutic drug monitoring of antiepileptic drugs : current status and future prospects. Expert Opin Drug Metab Toxicol [Internet]. 2020;00(00):1–12. Available at: https://doi.org/10.1080/17425255.2020.1724956
  14. Antunes MV, Charão MF, Linden R. Dried blood spots analysis with mass spectrometry: Potentials and pitfalls in therapeutic drug monitoring. Clin Biochem [Internet]. setembro de 2016;49(13–14):1035–46. Available at: https://doi.org/10.1016/j.clinbiochem.2016.05.004
  15. US-FDA 2018. USFDA, Guidance fo Industry: Bioanalytical method validation [Internet]. US Department of Health and HUman Service, US FDA, Center for Dug Evaluation and Resarch, Silver Spring. 2018. Available at: https://www.fda.gov/media/70858/download
  16. Linder C, Hansson A, Sadek S, Lars L, Pohanka A. Carbamazepine, lamotrigine, levetiracetam and valproic acid in dried blood spots with liquid chromatography tandem mass spectrometry; method development and validation. J Chromatogr B [Internet]. 2017; Available at: http://dx.doi.org/10.1016/j.jchromb.2017.11.005
  17. AbuRuz S, Al-Ghazawi M, Al-Hiari Y. A Simple Dried Blood Spot Assay for Therapeutic Drug Monitoring of Lamotrigine. Chromatographia [Internet]. 2010;71(11):1093–9. Available at: https://doi.org/10.1365/s10337-010-1569-y
  18. Kong ST, Lim S-H, Lee WB, Kumar PK, Wang HYS, Ng YLS, 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. https://doi.org/10.1371/journal.pone.0108190
  19. Pohanka A, Mahindi M, Masquelier MÈLE, Gustafsson LL, Beck O. Quantifi cation of valproic acid in dried blood spots. 2014;(August 2013):648–52. https://doi.org/10.3109/00365513.2014.933360
  20. Déglon J, Versace F, Lauer E, Widmer C, Mangin P, Thomas A, et al. Rapid LC-MS/MS quantification of the major benzodiazepines and their metabolites on dried blood spots using a simple and cost-effective sample pretreatment. Bioanalysis. junho de 2012;4(11):1337–50. https://doi.org/10.4155/bio.12.42
  21. Li W, Tse FLS. Dried blood spot sampling in combination with LC-MS/MS for quantitative analysis of small molecules. Biomed Chromatogr. janeiro de 2010;24(1):49–65. https://doi.org/10.1002/bmc.1367