Development and Validation of Lc-ms/ms Method for the Simultaneous Quantitative Analysis of Oxcarbazepine and Its Metabolite 10- Hydroxycarbazepine in K2edta Plasma

Objective: To develop and validate a sensitive, selective and reproducible method for the estimation of oxcarbazepine (OXC) and its pharmacologically active metabolite 10-hydroxycarbazepine (MHD) in human K2EDTA (Ethylene Diamine Tetra Acetic Acid) plasma. Methods: The drugs of interest were extracted by solid phase extraction technique, using HLB cartridges (30mg/1cc). Chromatographic separation was achieved in less than 3.0 minutes on reverse phase C8 column, using isocratic elution with acetonitrile-buffer solution (0.1% formic acid in water) in proportion of (70:30, v/v). Tandem mass spectrometer was used to detect the positive ions, using APCI (Atmospheric Pressure Chemical Ionization) probe. Results: The validated method was linear for OXC (0.032-7.044 μg/ml) and MHD (0.103-15.054 μg/ml) and the coefficient of correlation (r) for the drug and the metabolite were better than 0.99. The method was fully validated, complying FDA (Food and Drug Administration), EMEA (European Medicines Agency) guideline and recommendations of AAPS (American Association of Pharmaceutical Scientists) white papers demonstrating Selectivity, Sensitivity, Matrix factor, Precision and Accuracy, Linearity, Aqueous stability, Stability in biological matrix, Dilution Integrity, Reinjection Reproducibility, Recovery, Ruggedness and Extended Batch Verification. Conclusion: The validated method is applicable for Bioavailability/ Bioequivalence and Pharmacokinetic studies.