Development and Validation of LC-MS/MS Method for the Quantification of Oxcarbazepine in Human Plasma Using an Experimental Design

Abstract

Oxcarbazepine (OXB) is a newer antiepileptic drug indicated for the treatment of partial seizures as both monotherapy and combination therapy in adults and children with epilepsy. At present, determinations of OXB have been established by the use of HPLC-UV spectrometry, [1][2][3] or LC-APCI-MS mass-spectrometry. 4) However, none of these methods achieved the quick quantification and identification of OXB in a single run. Although methods for simultaneous determination of OXB and some of antiepileptic drugs has been described, 5-11) they were not particular interest since it takes too long chromatographic run time especially when applied to OXB, and had low sensitivity; it appeared that no assay existed for determination of the OXB using HPLC-MS/MS. The assay described here requires small mobile phase and sample volume, short chromatographic run and is sensitive, specific and fully validated.Optimization of an MS response includes methods to improve the abundance of a specific precursor or product ion, or yielding as comprehensive or selective measurements as possible (Table 1). Maximization of the signal or the signal-tonoise ratio (S/N) of a particular ion can be used for quantitative optimization, but choosing qualitative optimization criteria are less trivial.Optimization techniques of particular interest are factorial design and central composite design, for screening of the important parameters Placket-Burman design can be useful. Chemometric approaches for optimization of ESI or MS/MS parameters also occur in literature. 12,13) In this present study Plackett-Burman and factorial designs are employed for screening of essential parameters and optimization respectively. ExperimentalChemicals and Reagents OXB, and imipramine (Internal standard), were kindly provided by Glenmark Pharmaceuticals (Mumbai, India). HPLC-grade acetonitrile, methanol, formic acid and diethyl ether were purchased from Qualigens (Mumbai, India). Blank human plasma was harvested after 5 min centrifugation at 4000ϫg and stored at Ϫ20°C until use.Instrumentation The HPLC system consisted of an HP1100 binary pump, auto sampler, column oven, and online degasser (Agilent Technologies, Palo Alto, CA, U.S.A.). The analytical column used was reversedphase symmetry C18 (75 mmϫ4.6 mm i.d. 3.5 mm). A mobile phase of 10 mM ammonium formate/ACN (10 : 90, v/v) was pumped at a flow rate of 0.3 ml/min and the injected sample volume to whole over the method is 10 ml. The LC elute was introduced directly into an Applied Biosystems triple quadrupole mass spectrometer API 4000 (MDS-Sciex, Toronto Canada, software: Analyst 1.4.1) equipped with an electrospray ionization source. The triple quadrupole mass spectrometer was operated in the positive ion mode and the multiple reaction monitoring (MRM) chromatograms obtained were used for quantification. MRM transitions of m/z 253→208, m/z and 281→86 were respectively optimized for OXB and Imipramine. The 28Vol. 56, No. 1 A rapid tandem mass spectrometric (MS-MS) method for the quantification of Oxcarbazepine (O...

Electrochemical study and differential pulse voltammetric determination of oxcarbazepine and its main metabolite at a glassy carbon electrode

“…The analytical performance of the developed method was in agreement with other analytical methods. [18][19][20][21][22][23][24][25][26][27][28][29][30] Although the obtained results by this method were close to those of HPLC coupled with various detectors, these other methods suffer from the need for expensive equipment, and are time consuming with complex sample preparation.…”

“…16,17 Therefore, the level of the drug in blood serum is determined by the measuring its main metabolite (MHD). Several chromatographic methods for the determination of OX and MHD in biological uids have been developed, including gas chromatography assay, 18 high performance liquid chromatography-diode array detection, 19,20 high-performance liquid chromatography, 21 high-performance liquid chromatography (HPLC) with spectrophotometric detection, 22,23 microbore LC-MS/MS, 21 LC-APCI-MS mass spectrometry, 25 liquid chromatography-electrospray tandem mass spectrometry, [26][27][28][29] and capillary electrokinetic chromatography. 30 These methods require fastidious sample preparation, prolonged analysis time and expensive apparatus.…”

Quantitative bioanalytical and analytical method development of dibenzazepine derivative, carbamazepine: A review

“…A rapid tandem mass spectrometric (MS/MS) method for the quantification of OXCBZ in human plasma using imipramine as an internal standard has been developed and validated [25] . Plackett–Burman design was applied for screening of chromatographic and mass spectrometric factors.…”

Development and validation of a stability-indicating RP-LC method for the estimation of process-related impurities and degradation products of oxcarbazepine in pharmaceutical formulation

“…Estimation of oxcarbazepine and its metabolites in biological fluids by high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) has been performed [5][6][7][8][9][10][11]. Only one method was reported for the determination of oxcarbazepine and its related compounds by reverse-phase liquid chromatography (RP-LC) [12], but the method deals with only three impurities present in active pharmaceutical ingredient, and forced degradation studies were not performed properly.…”