Top

Forensic Science, Medicine and Pathology

Open Access 10-04-2024 | Lamotrigine | Original Article

Determination of lamotrigine in human plasma by HPLC-PDA. Application to forensic samples

Authors: Inés Sánchez-Sellero, Iván Álvarez-Freire, Pamela Cabarcos-Fernández, Lidia Janza-Candal, María Jesús Tabernero-Duque, Ana María Bermejo-Barrera

Published in: Forensic Science, Medicine and Pathology

Abstract

Purpose

Therapeutic drug monitoring of plasma lamotrigine (LTG) has customarily been carried out in order to prevent some its adverse effects. For forensic purposes, determination of LTG in plasma is an useful tool in cases of accidental overdose or suicidal attempts. Currently, there are several analytical methods available including some based on LC tandem mass spectrometry techniques, but simple and accessible LC-UV methods still can be useful for the purpose. Here we report on a new high-performance liquid chromatography method for the determination of lamotrigine in human plasma which has been developed and validated including selectivity, sensitivity, accuracy, precision and recovery studies.

Methods

Lamotrigine and the internal standard chloramphenicol were extracted from plasma using liquid-liquid extraction using small volumes of buffer and ethylacetate. Detection was monitored at 305.7 and 276.0 nm for lamotrigine and chloramphenicol, respectively.

Results

The method was linear concentration dependence within the range of 0.1–10 µg/ml, with a mean coefficient of correlation r = 0.993. The limit of detection (LOD) was 0.04 µg/ml and the limit of quantification (LOQ) was 0.1 µg/ml. Intra and interday precision values were lower than 9.0% at all concentrations studied. The intra and interday accuracy values ranged from − 7.6 to 10.1%. Recovery was found to be 98.9% or higher. The method here described was successfully applied to 11 postmortem blood samples received at the Forensic Sciences Institute of Santiago de Compostela (Spain).

Conclusion

A new HPLC method for the determination of lamotrigine in human plasma was developed and validated. A liquid-liquid extraction using small volumes of buffer and ethylacetate was optimized. The proposed method is suitable for forensic toxicological analysis.

Nevitt SJ, Sudell M, Cividini S, Marson AG, Tudur Smith C. Antiepileptic drug monotherapy for epilepsy: a network meta-analysis of individual participant data. Cochrane Database Syst Rev. 2022;4(4):CD011412. https://doi.org/10.1002/14651858.CD011412.pub4.CrossRefPubMed

Giri VP, Giri OP, Khan FA, Kumar N, Kumar A, Haque A. Valproic acid versus Lamotrigine as First-line Monotherapy in newly diagnosed idiopathic generalized Tonic-Clonic seizures in adults - a Randomized Controlled Trial. J Clin Diagn Res. 2016;10(7):FC01–4. https://doi.org/10.7860/JCDR/2016/16911.8121.CrossRefPubMedPubMedCentral

Betchel NT, Fariba KA, Saadabadi A. Lamotrigine. In: StatPearls, Treasure Island (FL): StatPearls Publishing;2023. https://www.ncbi.nlm.nih.gov/books/NBK470442/.

Smith MD, Metcalf CS, Wilcox KS. Pharmacotherapy of the epilepsies. In: Brunton LL, Hilal-Dandan R, Knollmann BC, editors. Goodman & Gilman’s: the pharmacological basis of therapeutics. 13th ed. New York: McGraw Hill; 2018. pp. 303–26.

Yang H, Zhang D, Mei S, Zhao Z. Simultaneous determination of plasma lamotrigine, lamotrigine N2-glucuronide and lamotrigine N2-oxide by UHPLC-MS/MS in epileptic patients. J Pharm Biomed Anal. 2022;220:115017. https://doi.org/10.1016/j.jpba.2022.115017.CrossRefPubMed

Contin M, Balboni M, Callegati E, Candela C, Albani F, Riva R, Baruzzi A. Simultaneous liquid chromatographic determination of lamotrigine, oxcarbazepine monohydroxy derivative and felbamate in plasma of patients with epilepsy. J Chromatogr B Analyt Technol Biomed Life Sci. 2005;828(1–2):113–7. https://doi.org/10.1016/j.jchromb.2005.09.009.CrossRefPubMed

Angelis-Stoforidis P, Morgan DJ, O’Brien TJ, Vajda FJ. Determination of lamotrigine in human plasma by high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl. 1999;727(1–2):113–8. https://doi.org/10.1016/s0378-4347(99)00043-2.CrossRefPubMed

Itabashi S, Bito R, Nishina M, Fukumoto M, Soda M, Doi M, Usui S, Kitaichi K. Determination of lamotrigine in human plasma using liquid chromatography-tandem mass spectrometry. Neuropsychopharmacol Rep. 2019;39(1):48–55. https://doi.org/10.1002/npr2.12045.CrossRefPubMedPubMedCentral

Clark A, St Clair B. Severe cutaneous adverse reactions Associated with High-Dose Lamotrigine for Mood disorders: a Case Series. Innov Pharm. 2022;13(2). https://doi.org/10.24926/iip.v13i2.4541.

10.

Barbosa NR, Mídio AF. Validated high-performance liquid chromatographic method for the determination of lamotrigine in human plasma. J Chromatogr B Biomed Sci Appl. 2000;741(2):289–93. https://doi.org/10.1016/s0378-4347(00)00102-x.CrossRefPubMed

11.

Greiner-Sosanko E, Lower DR, Virji MA, Krasowski MD. Simultaneous determination of lamotrigine, zonisamide, and carbamazepine in human plasma by high-performance liquid chromatography. Biomed Chromatogr. 2007;21(3):225–8. https://doi.org/10.1002/bmc.753.CrossRefPubMedPubMedCentral

12.

Greiner-Sosanko E, Giannoutsos S, Lower DR, Virji MA, Krasowski MD. Drug monitoring: simultaneous analysis of lamotrigine, oxcarbazepine, 10-hydroxycarbazepine, and zonisamide by HPLC-UV and a rapid GC method using a nitrogen-phosphorus detector for levetiracetam. J Chromatogr Sci. 2007;45(9):616–22. https://doi.org/10.1093/chromsci/45.9.616.CrossRefPubMed

13.

Saracino MA, Bugamelli F, Conti M, Amore M, Raggi MA. Rapid HPLC analysis of the antiepileptic lamotrigine and its metabolites in human plasma. J Sep Sci. 2007;30(14):2249–55. https://doi.org/10.1002/jssc.200700110.CrossRefPubMed

14.

Ventura S, Rodrigues M, Pousinho S, Falcão A, Alves G. An easy-to-use liquid chromatography assay for the analysis of lamotrigine in rat plasma and brain samples using microextraction by packed sorbent: application to a pharmacokinetic study. J Chromatogr B Analyt Technol Biomed Life Sci. 2016;1035:67–75. https://doi.org/10.1016/j.jchromb.2016.09.032.CrossRefPubMed

15.

Ranjbar S, Daryasari AP, Soleimani M. Ionic liquid-based Dispersive Liquid-Liquid Microextraction for the Simultaneous Determination of Carbamazepine and Lamotrigine in Biological samples. Acta Chim Slov. 2020;67(3):748–56.CrossRefPubMed

16.

Abou-Taleb NH, El-Sherbiny DT, El-Enany NM, El-Subbagh HI. A new grey relational analysis application in analytical chemistry: natural deep eutectic solvent as a green extractant for HPLC determination of lamotrigine in plasma. Microchem J. 2022;172A:106918. https://doi.org/10.1016/j.microc.2021.106918.CrossRef

17.

Sabença R, Bicker J, Silva R, Carona A, Silva A, Santana I, Sales F, Falcão A, Fortuna A. Development and application of an HPLC-DAD technique for human plasma concentration monitoring of perampanel and lamotrigine in drug-resistant epileptic patients. J Chromatogr B Analyt Technol Biomed Life Sci. 2021;1162:122491. https://doi.org/10.1016/j.jchromb.2020.122491.CrossRefPubMed

18.

Madhavaram H, Woollard GA, Couch RA. Measurement of serum lamotrigine by high performance liquid chromatography using a phenyltriazine as internal standard. Drug Test Anal. 2012;4(2):136–9. https://doi.org/10.1002/dta.255.CrossRefPubMed

19.

Serralheiro A, Alves G, Fortuna A, Rocha M, Falcão A, First. HPLC-UV method for rapid and simultaneous quantification of phenobarbital, primidone, phenytoin, carbamazepine, carbamazepine-10,11-epoxide, 10,11-trans-dihydroxy-10,11-dihydrocarbamazepine, lamotrigine, oxcarbazepine and licarbazepine in human plasma. J Chromatogr B Analyt Technol Biomed Life Sci. 2013;925:1–9. https://doi.org/10.1016/j.jchromb.2013.02.026.CrossRefPubMed

20.

Deeb S, McKeown DA, Torrance HJ, Wylie FM, Logan BK, Scott KS. Simultaneous analysis of 22 antiepileptic drugs in postmortem blood, serum and plasma using LC-MS-MS with a focus on their role in forensic cases. J Anal Toxicol. 2014;38(8):485–94. https://doi.org/10.1093/jat/bku070.CrossRefPubMed

21.

Hotha KK, Kumar SS, Bharathi DV, Venkateswarulu V. Rapid and sensitive LC-MS/MS method for quantification of lamotrigine in human plasma: application to a human pharmacokinetic study. Biomed Chromatogr. 2012;26(4):491–6. https://doi.org/10.1002/bmc.1692.CrossRefPubMed

22.

Palte MJ, Basu SS, Dahlin JL, Gencheva R, Mason D, Jarolim P, Petrides AK. Development and validation of an Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry Method for the Concurrent Measurement of Gabapentin, Lamotrigine, Levetiracetam, Monohydroxy Derivative of Oxcarbazepine, and Zonisamide concentrations in serum in a clinical setting. Ther Drug Monit. 2018;40(4):469–76. https://doi.org/10.1097/FTD.0000000000000516.CrossRefPubMedPubMedCentral

23.

Shi M, Jiang Q, Lyu Q, Yuan Z, Deng L, Yin L. A LC-MS3 strategy to determine lamotrigine by Q-Q-trap tandem mass spectrometry coupled with triple stage fragmentation to enhance sensitivity and selectivity. Anal Methods. 2021;13(38):4478–84. https://doi.org/10.1039/D1AY01372F.CrossRefPubMed

24.

Zhou S, Li R, Chen Z, Ren R, Wang X, Dai Q, Wen D, Guan Y, Zhang X, Tang S, Zhou L, Huang M. LC-MS/MS quantification of levetiracetam, lamotrigine and 10-hydroxycarbazepine in TDM of epileptic patients. Biomed Chromatogr. 2022;36(8):e5393. https://doi.org/10.1002/bmc.5393.CrossRefPubMed

25.

Nikolaou P, Papoutsis I, Dona A, Spiliopoulou C, Athanaselis S. Development and validation of a GC/MS method for the simultaneous determination of levetiracetam and lamotrigine in whole blood. J Pharm Biomed Anal. 2015;102:25–32. https://doi.org/10.1016/j.jpba.2014.08.034.CrossRefPubMed

26.

Food and Drug Administration. Bioanalytical Method Validation. Guidance for Industry. Biopharmaceutics. 2018. https://www.fda.gov/files/drugs/published/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf.

27.

Bugamelli F, Sabbioni C, Mandrioli R, Kenndler E, Albani F, Raggi MA. Simultaneous analysis of six antiepileptic drugs and two selected metabolites in human plasma by liquid chromatography after solid-phase extraction. Anal Chim Acta. 2002;472(1–2):1–10. https://doi.org/10.1016/S0003-2670(02)00980-7.CrossRef

28.

Ventura S, Rodrigues M, Pousinho S, Falcão A, Alves G. Determination of lamotrigine in human plasma and saliva using microextraction by packed sorbent and high performance liquid chromatography–diode array detection: an innovative bioanalytical tool for therapeutic drug monitoring. Microchem J. 2017;130:221–8. https://doi.org/10.1016/J.MICROC.2016.09.007.CrossRef

29.

Pricone MG, King CV, Drummer OH, Opeskin K, McIntyre IM. Postmortem investigation of lamotrigine concentrations. J Forensic Sci. 2000;45(1):11–5.CrossRefPubMed

30.

Levine B, Jufer RA, Smialek JE. Lamotrigine distribution in two postmortem cases. J Anal Toxicol. 2000;24(7):635–37. https://doi.org/10.1093/jat/24.7.635.CrossRefPubMed

Title: Determination of lamotrigine in human plasma by HPLC-PDA. Application to forensic samples
Authors: Inés Sánchez-Sellero
Iván Álvarez-Freire
Pamela Cabarcos-Fernández
Lidia Janza-Candal
María Jesús Tabernero-Duque
Ana María Bermejo-Barrera
Publication date: 10-04-2024
Publisher: Springer US
Keywords: Lamotrigine
Chloramphenicol
Published in: Forensic Science, Medicine and Pathology
Print ISSN: 1547-769X
Electronic ISSN: 1556-2891
DOI: https://doi.org/10.1007/s12024-024-00812-9

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Determination of lamotrigine in human plasma by HPLC-PDA. Application to forensic samples

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content