Objective: The objective of this research was to develop and validate a simple, sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for quantification of Topiramate in human plasma.

Method: The analytical method consists of solid phase extraction of plasma sample followed by the determination of Topiramate by a LC–MS/MS. The analyte was separated on Advance Hypurity C18 (50 x 4.6 mm, 5µ) column with an isocratic mobile phase of Acetonitrile and 2mM ammonium acetate buffer in the volume ratio of 80:20 v/v at a flow rate of 0.8 mL/min.     Results: Protonated ions formed by a turbo ion spray in a negative mode were used to detect analyte and internal standard (IS). The MS/MS detection was made by monitoring the fragmentation of m/z 338.00→77.50 for Topiramate and m/z 350.40→90.10 for internal standard on a mass spectrometer. The method was validated over the concentration range of 20.721 ng/mL to 605.656 ng/mL for Topiramate in human plasma with correlation coefficient greater than 0.99.

Conclusion: The developed method was successfully applied for analyzing Topiramate in plasma samples for a bioequivalence study with healthy volunteers.


Key words: Topiramate; Validation; Analysis; Bioavailability; LC-MS/MS.


1. Shank RP, Gardocki JF, Streeter AJ, Maryanoff BE. An Overview of the Preclinical Aspects of Topiramate: Pharmacology, Pharmacokinetics, and Mechanism of Action. Epilepsia 2000; 41(1):3.
2. Guerrini R, Carpay J. Topiramate monotherapy as broad-spectrum antiepileptic drug in a naturalistic clinical setting. Seizure 2005; 14:371.
3. Ajlouni SA, Shorman A, Daoud AS. The efficacy and side effects of topiramate on refractory epilepsy in infants and young children: A multi-center clinical trial. Seizure 2005; 14:459.
4. Styslo-Zalasik M, Weiyong Li. Determination of topiramate and its degradation product in liquid oral solutions by high performance liquid chromatography with a chemiluminescent nitrogen detector. J. Pharm. Biomed. Anal. 2005; 37(3):529.
5. Bahrami G, Mohammadi B. A novel high sensitivity HPLC assay for topiramate, using 4-chloro-7-nitrobenzofurazan as pre-column fluorescence derivatizing agent. J. Chromatogr. B 2007; 850:400.
6. Gh. Bahrami, Sh. Mirzaeei, Kiani A. Sensitive analytical method for Topiramate in human serum by HPLC with pre-column fluorescent derivatization and its application in human pharmacokinetic studies. J. Chromatogr. B 2004; 813:175.
7. Bahrami B, Mirzaeei S, Mohammadi B, Kiani A. High performance liquid chromatographic determination of topiramate in human serum using UV detection. J. Chromatogr. B 2005; 822:322.
8. Mercolini L, Mandrioli R, Amore M, Raggi MA. Simultaneous HPLC-F analysis of three recent antiepileptic drugs in human plasma. J. Pharm. Biomed. Anal. 2010; 53:62.
9. Mandrioli R, Musenga A, Kenndler E, Donno M, Amore M, Raggi MA. Determination of topiramate in human plasma by capillary electrophoresis with indirect UV detection. J. Pharm. Biomed. Anal. 2010; 53:1319.
10. Klockow-Beck A, Nick A, Schaufelberger D. Determination of the inorganic degradation products sulfate and sulfamate in the antiepileptic drug topiramate by capillary electrophoresis. J. Chromagr. B: Biome. Sci. Appl. 1998; 720:141.
11. Carl E. Wolf, C. Richard Crooks, Poklis A. Rapid Gas Chromatographic Procedure for the Determination of Topiramate in Serum. J. Anal. Toxicol. 2000; 24:661.
12. Matar KM, Therapeutic drug monitoring of topiramate by liquid chromatography-tandem mass spectrometry. Clin. Chim. Acta 2010; 411 (9-10):729.
13. Pineyro-Lopez A, Pineyro-Garza E, Gomez-Silva M. Bioequivalence of single 100-mg doses of two oral formulations of topiramate: An open-label, randomized-sequence, two-period crossover study in healthy adult male Mexican volunteers. Clin. Therap 2009; 31:411.
14. Contin M, Riva R, Albani F, Baruzzi A. Simple and rapid liquid chromatographic–turbo ion spray mass spectrometric determination of topiramate in human plasma. J. Chromatogr. B: Biomed. Sci. Appl. 2001; 761:133.
15. Goswami D, Kumar A, Khuroo AH, Monif T, Rab S. Bioanalytical LC-MS/MS method validation for plasma determination of topiramate in healthy Indian volunteers. Biomed. Chromatogr. 2009: 23:1227.
16. Park JH, Park YS. Determination of plasma topiramate concentration using LC-MS/MS for pharmacokinetic and bioequivalence studies in healthy Korean volunteers. Biomed. Chromatogr. 2008, 22:822.
17. Malka B, Stefan S, Nina I. Analysis of Topiramate and Its Metabolites in Plasma and Urine of Healthy Subjects and Patients with Epilepsy by Use of a Novel Liquid Chromatography-Mass Spectrometry Assay. Therapeu. Drug Monit, 2003; 25:314.
18. Guidance for Industry, Bioanalytical Method validation, CDER, May 2001.
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How to Cite
KUMAR , D. G. “ESTIMATION OF TOPIRAMATE IN HUMAN PLASMA USING LC-MS/MS METHOD”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 6, no. 7, Aug. 2013, pp. 217-20,