Enhanced Detection of Lamotrigine Through HPLC Methodology in Bioanalysis
Article Sidebar
Main Article Content
Abstract
Background: Lamotrigine, an anticonvulsant drug, requires sensitive detection methods in biological and pharmaceutical matrices. Current methods struggle with interference and sensitivity, necessitating the development of improved analytical techniques.
Objective: This study aims to develop and validate a highly sensitive HPLC method enhanced by salicylaldehyde derivatization for detecting lamotrigine in pharmaceutical and biological samples, and to differentiate it from Piracetam when co-administered.
Methods: A reverse-phase HPLC method was employed, utilizing salicylaldehyde as a derivatizing agent to induce a bathochromic shift in lamotrigine from 315 nm to 415 nm. Optimization of the derivatization reaction included adjustments of pH, reagent concentration, temperature, and time. The standard addition method was applied to evaluate the recovery rates from spiked samples.
Results: The optimized method demonstrated a linear calibration range of 1-5 µg/ml with a determination coefficient (R²) of 0.998. Recovery of lamotrigine from pharmaceutical preparations averaged 96.5%, while recovery from deprotonated serum and urine samples of healthy volunteers was 96% with a relative standard deviation (RSD) of 6%.
Conclusion: The developed method offers a robust and highly sensitive approach to detect and distinguish lamotrigine in the presence of Piracetam, proving effective for both pharmaceutical and biological sample analysis.
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
References
Biddlecombe R, Dean K, Smith C, Jeal S. Validation of a radioimmunoassay for the determination of human plasma concentrations of lamotrigine. J Pharm Biomed Anal. 1990;8(6):691-4.
Yamashita S, Furuno K, Kawasaki H, Gomita Y, Yoshinaga H, Yamatogi Y, Ohtahara S. Simple and rapid analysis of lamotrigine, a novel antiepileptic, in human serum by high-performance liquid chromatography using a solid-phase extraction technique. J Chromatogr B Biomed Sci Appl. 1995;670:354-7.
Fraser AD, MacNeil W, Isner AF, Camfield PR. Lamotrigine analysis in serum by high-performance liquid chromatography. J Ther Drug Monit. 1995;17(2):174-8.
Londero D, Greco PL. New micromethod for the determination of lamotrigine in human plasma by high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl. 1997;691:139-44.
Sallustio BC, Morris RG. High-performance liquid chromatography quantitation of plasma lamotrigine concentrations: application measuring through concentrations in patients with epilepsy. J Ther Drug Monit. 1997;19(6):688-93.
Hart AP, Mazarr-Proo S, Blackwell W, Dasgupta A. A rapid cost-effective high-performance liquid chromatographic (HPLC) assay of serum lamotrigine after liquid-liquid extraction and using HPLC conditions routinely used for analysis of barbiturates. J Ther Drug Monit. 1997;19(4):431-5.
Lensmeyer GL, Gidal BE, Wiebe DA. Optimized high-performance liquid chromatographic method for determination of lamotrigine in serum with concomitant determination of phenytoin, carbamazepine, and carbamazepine epoxide. J Ther Drug Monit. 1997;19(3):292-300.
Cooper J, Shearsby N, Taylor J, Sheung CF. Simultaneous determination of lamotrigine and its glucuronide and methylated metabolites in human plasma by automated sequential trace enrichment of dialysates and gradient high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl. 1997;702:227-33.
Matar K, Nicholls P, Bawazir S, Al-Hassan M, Tekle A. A rapid liquid chromatographic method for the determination of lamotrigine in plasma. J Pharm Biomed Anal. 1998;17(3):525-31.
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;727:113-8.
Vidal E, Pascual C, Pou L. Determination of lamotrigine in human serum by liquid chromatography. J Chromatogr B Biomed Sci Appl. 1999;736:295-8.
Torra M, Rodamilans M, Arroyo S, Corbella J. Optimized procedure for lamotrigine analysis in serum by high-performance liquid chromatography without interferences from other frequently coadministered anticonvulsants. J Ther Drug Monit. 2000;22(6):621-5.
Barbosa NR, Mı́dio AF. Validated high-performance liquid chromatographic method for the determination of Lamotrigine in human plasma. J Chromatogr B Biomed Sci Appl. 2000;741:289-93.
Croci D, Salmaggi A, de Grazia U, Bernardi G. New high-performance liquid chromatographic method for plasma/serum analysis of Lamotrigine. J Ther Drug Monit. 2001;23(6):665-8.
Cheng CL, Chou CH, Hu OY. Determination of Lamotrigine in small volumes of plasma by high-performance liquid chromatography. J Chromatogr B. 2005;817:199-206.
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. 2005;828:113-7.
Patil KM, Bodhankar SL. High-performance thin-layer chromatographic determination of Lamotrigine in serum. J Chromatogr B. 2005;823:152-7.
Emami J, Ghassami N, Ahmadi F. Development and validation of a new HPLC method for determination of Lamotrigine and related compounds in tablet formulations. J Pharm Biomed Anal. 2006;40:999-1005.
Greiner C, Haen E. Development of a simple column-switching high-performance liquid chromatography (HPLC) method for rapid and simultaneous routine serum monitoring of Lamotrigine, oxcarbazepine and 10-monohydroxycarbazepine (MHD). J Chromatogr B. 2007;854:338-44.
Vermeij T, Edelbroek P. Robust isocratic high performance liquid chromatographic method for simultaneous determination of seven antiepileptic drugs including lamotrigine, oxcarbazepine and zonisamide in serum after solid-phase extraction. J Chromatogr B. 2007;857:40-6.
Calvo ME, Renedo OD, Martínez MJ. Determination of lamotrigine by adsorptive stripping voltammetry using silver nanoparticle-modified carbon screen-printed electrodes. Talanta. 2007;74:59-64.
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;21:225-8.
Bompadre S, Tagliabracci A, Battino M, Giorgetti R. Determination of lamotrigine in whole blood with online solid phase extraction. J Chromatogr B. 2008;863:177-80.
Del Rosario Brunetto M, Contreras Y, Delgado Y, Gallignani M, Estela JM, Martin VC. Development and validation of a rapid column-switching high-performance liquid chromatographic method for the determination of lamotrigine in human serum. J Chromatogr Sci. 2009;47:478-84.
Hosseini MH, Alizadeh N, Khanchi AR. Solubility analysis of clozapine and lamotrigine in supercritical carbon dioxide using static system. J Supercrit Fluids. 2010;52:30-5.
Rivas N, Zarzuelo A, López F. Optimisation of a high-efficiency liquid chromatography technique for measuring lamotrigine in human plasma. Farmacia Hospitalaria (English Edition). 2010;34:85-9.
Antonilli L, Brusadin V, Filipponi F, Guglielmi R, Nencini P. Development and validation of an analytical method based on high performance thin layer chromatography for the simultaneous determination of lamotrigine, zonisamide and levetiracetam in human plasma. J Pharm Biomed Anal. 2011;56:763-70.
Saberi RS, Shahrokhian S. Highly sensitive voltammetric determination of lamotrigine at highly oriented pyrolytic graphite electrode. J Bioelectrochemistry. 2012;84:38-43.
Zhiyan L, Haitao L, Yang S, Fang W, Huaiping T, Ajing X. An HPLC Method for Simultaneous Determination of Plasma Concentration of Lamotrigine and Oxcarbazepine Active Metabolite and Its Application in Children with Epilepsy. J Pediatric Pharmacy. 2013;2:003.
Najib FM, Aziz KH. Spectrophotometric determination of lamotrigine in pharmaceutical preparations and urine samples using bromothymol blue and bromophenol blue. Malasian J Analytical Sciences. 2013;17:310-25.
Gholivand MB, Malekzadeh G, Torkashvand M. Determination of lamotrigine by using molecularly imprinted polymer–carbon paste electrode. J Electroanal Chem. 2013;692:9-16.
Bhat K. Determination of lamotrigine and felbamate in pediatric plasma samples by reverse phase high-performance liquid chromatography. Ind J Pharm Edu Res. 2013;2:47-52.
Abolmaali SS, Tamaddon AM, Samani SM. Application of rapid and simple liquid chromatography method for determination of bioequivalence of generic lamotrigine tablets in healthy Iranian volunteers. Trends Pharm Sci. 2015;1:159-66.
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.
Jebabli N, Gaïes E, El Jebari H, Charfi R, Lakhal M, Klouz A, Trabelsi S, Salouage I. Development and validation of a new HPLC method for determination of Lamotrigine and clinical application. La Tunisie Medicale. 2015;93:565-68.
Bojdi MK, Behbahani M, Hesam G, Mashhadizadeh MH. Application of magnetic lamotrigine-imprinted polymer nanoparticles as an electrochemical sensor for trace determination of lamotrigine in biological samples. RSC Advances. 2016;6:32374-80.
Chouinard G, Annable L, Ross-Chouinard A, Olivier M, Fontaine F. Piracetam in elderly psychiatric patients with mild diffuse cerebral impairment. Psychopharmacology. 1983;81:100-106.
Da Silva CM, da Silva DL, Modolo LV, Alves RB, de Resende MA, Martins CV, Fátima  de. Schiff bases: A short review of their antimicrobial activities. Advanced Research. 2011;2:1-8.
Foldvary N, Perry M, Lee J, Dinner D, Morris HH. The effects of lamotrigine on sleep in patients with epilepsy. Epilepsia. 2001;42:1569-73.
Hakkarainen H, Hakamies L. Piracetam in the treatment of post-concussional syndrome. European Neurology. 1978;17:50-55.
Janeš D, Kreft S. Salicylaldehyde is a characteristic aroma component of buckwheat groats. Food Chemistry. 2008;109:293-298.
Leach MJ, Marden CM, Miller AA. Pharmacological studies on lamotrigine, a novel potential antiepileptic drug. Epilepsia. 1986;27:490-497.
Stein D, Zungu-Dirwayi N, Linden G, Seedat S. Pharmacotherapy for post-traumatic stress disorder (PTSD). The Cochrane Library. 2000.
Surges R, Walker MC. A case of late-onset Kleine–Levin syndrome responding to lamotrigine. Sleep Medicine. 2009;10:394.
Castel-Branco M, Almeida A, Falcao A, Macedo T, Caramona M, Lopez F. Lamotrigine analysis in blood and brain by high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl. 2001;755:119-27.
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. 2005;828:113-17.
Greiner C, Haen E. Development of a simple column-switching high-performance liquid chromatography (HPLC) method for rapid and simultaneous routine serum monitoring of lamotrigine, oxcarbazepine and 10-monohydroxycarbazepine (MHD). J Chromatogr B. 2007;854:338-44.
Abolmaali SS, Tamaddon AM, Dinarvand R. A review of therapeutic challenges and achievements of methotrexate delivery systems for treatment of cancer and rheumatoid arthritis. Cancer Chemother Pharmacol. 2013;71:1115-30.