Simultaneous Determination of Carbamazepine and Carbamazepine-10,11-epoxide in Different Biological Matrices by LC-MS/MS

Open access


An uncomplicated, sensitive liquid chromatography linked to mass spectrometry (LC/MS) for evaluation of carbamazepine and carbamazepine-10,11-epoxide (its metabolite) in human plasma, human saliva, rat plasma, and rabbit plasma was developed. Analyses were conducted on a Zorbax SB-C18, 100 mm × 3 mm ID, 3.5 μm column, at a column temperature of 40 ºC. The mobile phase was comprised of 0.1% formic acid in water and methanol in a 35 : 65 (v/v) ratio, with a flow rate of 0.4 mL/min. Lacosamide was utilized as internal standard. Under these chromatographic conditions, the retention times of lacosamide, carbamazepine-10,11-epoxide, and carbamazepine were 1.4 min, 1.6 min, and 2.2 min, respectively. The quantification of the analytes was performed using multiple reaction monitoring, with the use of a triple quadrupole mass spectrometer with electrospray positive ionization. The monitored ions were m/z 194 derived from m/z 237 for carbamazepine, m/z 180 derived from m/z 253 for carbamazepine-10,11-epoxide, and m/z 108 derived from m/z 251 for lacosamide. The samples were prepared by protein precipitation from 0.2 mL of plasma/saliva using 0.6 mL of internal standard solution in methanol. Calibration curves were constructed over the ranges 1.1–17.6 µg/mL and 0.23–5.47 µg/mL for carbamazepine and carbamazepine-epoxide, respectively. The coefficients of determination obtained by using a weighted (1/x) linear regression were greater than 0.994. The reported LC-MS/MS method was applied to preclinical pharmacokinetic studies and therapeutic drug monitoring.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • 1. Ettinger AB Argoff CE. Use of Antiepileptic Drugs for Nonepileptic Conditions: Psychiatric Disorders and Chronic Pain. Neurotherapeutics. 2007;4:75-83.

  • 2. Brodie MJ. Antiepileptic drug therapy the story so far. Seizure. 2010;19:650-655.

  • 3. Kerr BM Thummel KE Wurden CJ et al. Human liver carbamazepine metabolism. Biochem Pharmacol. 1994;47:1969-1979.

  • 4. Johannessen SI Landmark CJ. Antiepileptic drug interactions - principles and clinical implications. Curr Neuropharmacol. 2010;8:254-267.

  • 5. Patsalos PN Berry DJ Bourgeois BFD et al. Antiepileptic drugs - Best practice guidelines for therapeutic drug monitoring: A position paper by the subcommission on therapeutic drug monitoring ILAE Commission on Therapeutic Strategies. Epilepsia. 2008;49:1239-1276.

  • 6. Johannessen SI Battino D Berry DJ et al. Therapeutic drug monitoring of the newer antiepileptic drugs. Ther Drug Monit. 2003;25:347-363.

  • 7. Neels HM Sierens AC Naelaerts K Scharpé SL Hatfield GM Lambert WE. Therapeutic drug monitoring of old and newer anti-epileptic drugs. Vol. 42 Clinical Chemistry and Laboratory Medicine; 2004. p. 1228-1255.

  • 8. Kim K-B Seo K-A Kim S-E Bae SK Kim D-H Shin J-G. Simple and accurate quantitative analysis of ten antiepileptic drugs in human plasma by liquid chromatography/tandem mass spectrometry. J Pharm Biomed Anal. 2011;56:771-777.

  • 9. Zhu Y Chiang H Wulster-Radcliffe M et al. Liquid chromatography/tandem mass spectrometry for the determination of carbamazepine and its main metabolite in rat plasma utilizing an automated blood sampling system. J Pharm Biomed Anal. 2005;38:119-125.

  • 10. Breton H Cociglio M Bressolle F Peyriere H Blayac JP Hillaire-Buys D. Liquid chromatography-electrospray mass spectrometry determination of carbamazepine oxcarbazepine and eight of their metabolites in human plasma. J Chromatogr B Anal Technol Biomed Life Sci. 2005;828:80-90.

  • 11. Subramanian M Birnbaum AK Remmel RP. High-Speed Simultaneous Determination of Nine Antiepileptic Drugs Using Liquid Chromatography-Mass Spectrometry. Ther Drug Monit. 2008;30:347-356.

  • 12. Theodore WH Narang PK Holmes MD Reeves P Nice FJ. Carbamazepine and its epoxide: Relation of plasma levels to toxicity and seizure control. Ann Neurol. 1989;25:194-196.

  • 13. Burianová I Bořecká K. Routine therapeutic monitoring of the active metabolite of carbamazepine: Is it really necessary? Clin Biochem. 2015;48:866-869.

  • 14. Krasniqi S Zeitlinger M Bauer S. Comparison between the immunoassay and high performance liquid chromatography for therapeutic monitoring of carbamazepine and phenytoine. Biochem Medica. 2010;341-349.

  • 15. Ferreira A Rodrigues M Oliveira P et al. Liquid chromatographic assay based on microextraction by packed sorbent for therapeutic drug monitoring of carbamazepine lamotrigine oxcarbazepine phenobarbital phenytoin and the active metabolites carbamazepine-1011-epoxide and licarbazepine. J Chromatogr B Anal Technol Biomed Life Sci. 2014;971:20-29.

  • 16. Shokry E Villanelli F Malvagia 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;109:164-170.

  • 17. Đorđević S Kilibarda V Stojanović T. Determination of carbamazepine in serum and saliva samples by high performance liquid chromatography with ultraviolet detection Odre đ ivanje karbamazepina u uzorcima seruma i salive primenom te č ne hromatografije visokih performansi sa ultravioletnom de. Vojn Pregl. 2009;66:347-352.

  • 18. Krasowski MD McMillin GA. Advances in anti-epileptic drug testing. Clin Chim Acta. 2014;436:224-236.

  • 19. Beig A Dahan A. Quantification of carbamazepine and its 1011-epoxide metabolite in rat plasma by UPLC-UV and application to pharmacokinetic study. Biomed Chromatogr. 2014;28:934-938.

  • 20. Ates Z Oezden T Oezilhan S Toptan S. Simultaneous determination of carbamazepine and its active metabolite carbamazepine-1011-epoxide in human plasma by UPLC. Chromatographia. 2007;66:S123-S127.

  • 21. Nunes LAS Mussavira S Bindhu OS. Clinical and diagnostic utility of saliva as a non-invasive diagnostic fluid: a systematic review. Biochem Med (Zagreb). 2015;25:177-192.

Journal information
All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 433 293 12
PDF Downloads 200 118 6