A VALIDATED LC-MS/MS METHOD FOR PHARMACOKINETIC STUDY OF BRIVARACETAM IN HEALTHY RABBITS
Objective: A liquid chromatography-tandem mass spectrophotometric (LCâ€“MS/MS) method was developed for quantification of brivaracetam in rabit plasma employing liquid-liquid extraction with ethyl acetate.
Methods: Developed method was validated for specificity, precision, accuracy, recovery, and stability characteristics. Chromatographic separation was achieved on Chromolith C18column (100 mmx4.6 mmx5 Âµm) with 0.1% formic acid, adjusted to pH 3.2 as an isocratic mobile phase with a flow rate of 1.0 ml/min. the developed method was applied to assess pharmacokinetics parameters like Cmax, Tmax, t1/2 and AUC of brivaracetam in healthy rabbits.
Results: The developed method was linear over the range of 0.16 to 8Âµg/ml. The regression equation for the analysis was Y=0.0053x+0.0018 with coefficient of correction (r2) = 0.998. The % mean recovery for brivaracetam was found to be between 95.7% to 106.5%. The mean intraday and inter-day precision of the method was found to be 0.77 to 3.72% for quality control standards. Brivaracetam showed Tmax of 1.025Â±0.061 and mean Cmax, AUC0Â®t andAUC0Â®a for Test formulation is 92.7Â±4.4, 496.21Â±26.4 and 504.20Â±30.68 respectively.
Conclusion: A highly specific, rugged and rapid method with sufficiently low LLOQ was developed for analysis of routine samples of a single dose or multiple dose pharmacokinetic studies with any marketing formulation of brivaracetam.
2. Brophy GM, Bell R, Claassen J. Guidelines for the evaluation and management of status epilepticus. Neurocrit Care 2012;17:3-23.
3. Varelas PN, Spanaki MV, Mirski MA. Status epilepticus: an update. Curr Neurol Neurosci Rep 2013;13:357.
4. Riviello JJ Jr, Claassen J, LaRoche SM. Treatment of status epilepticus: an international survey of experts. Neurocrit Care 2013;18:193-200.
5. Mula M. Brivaracetam for the treatment of epilepsy in adults. Expert Rev Neurother 2014;14:361-5.
6. Matagne A, Margineanu DG, Kenda B. Anti-convulsive and antiepileptic properties of brivaracetam (ucb 34714), a high-affinity ligand for the synaptic vesicle protein, SV2A. Br J Pharmacol 2008;154:1662-71.
7. Kenda B, Matagne A, Talaga P, Pasau P, Differding B, Lallemand B, et al. Discovery of 4-substituted pyrrolidone butanamides as new agents with significant antiepileptic activity. J Med Chem 2004;47:530-49.
8. Lynch B, Lambeng N, Nocka K, Kensel P, Bajjalieh S, Matagne A, et al. The synaptic vesicle protein SV2A is the binding site for the antiepileptic drug levetiracetam. Proc Natl Acad Sci 2004;101:9861-6.
9. Jacobs P, Ridder L, Ruijken M, Rosing H, Jager G, Beijnen J, et al. Identification of drug metabolites in human plasma or serum integrating metabolite prediction, LC-HRMS and untargeted data processing. Bioanalysis 2013;5:2115-28.
10. Maria L, Sargentini M, Pascal E. Pharmacokinetics and metabolism of 14C-brivaracetam, a novel SV2A ligand, in healthy subjects. Drug Metabolism 2008;36:36-45.
11. Nicolas J, Chanteux H, Rosa M, Watanabe S, Stockis A. Effect of gemfibrozil on the metabolism of brivaracetam in vitro and in human subjects. Drug Metab Dispos 2012;40:1466-72.
12. Zhang K, Li H, Myung K, Cho C, James C. Expanding metabolism for total biosynthesis of the nonnatural amino acid L-homoalanine. Proc Natl Acad Sci United 2010;107:6234â€“9.
13. Nicolas J, Hannestad J, Holden D, Kervyn S, Nabulsi N, Tytgat D, et al. Brivaracetam, a selective high-affinity synaptic vesicle protein 2A (SV2A) ligand with preclinical evidence of high brain permeability and fast onset of action. Epilepsia 2015;57:201-9.
14. ICH guidelines for the stability of new drug substances and products. Q1A (R2) ICH, Geneva; 2005. p. 1-13.
15. ICH guidelines for validation of analytical procedures: text and methodology. Q2 (R1) ICH, Geneva; 2005. p. 1-14.
16. Darshan B, Rajkamal B. A UPLC-MS/MS method development and validation for the estimation of sofosbuvir from human plasma. Int J Appl Pharm 2017;9:30-6.
17. Srinidhi M, Mushabbar Basha MD, V Raj Kumar, Rajendra Kumar J. Stability indicating RP-HPLC method development and validation for the estimation of sumatriptan in bulk and pharmaceutical dosage form. J Appl Pharm Sci 2016;6:20-5.