Simultaneous quantitative analysis of oxcarbazepine and 10,11-dihydro-10-hydroxycarbamazepine in human plasma by liquid chromatography-electrospray tandem mass spectrometry

J Pharm Biomed Anal. 2007 Oct 18;45(2):304-11. doi: 10.1016/j.jpba.2007.08.001. Epub 2007 Aug 9.

Abstract

A fast and sensitive method to quantify oxcarbazepine (OXC) and its active metabolite, 10,11-dihydro-10-hydroxycarbamazepine (MHD) in human plasma using HPLC-MS/MS has been developed. The method involved liquid-liquid extraction (LLE), with diethyl ether-diclhoromethane (60:40v/v) using deuterade carbamazepine (d10-carbamazepine) as internal standard (IS). The analytes and IS were separated using an isocratic mobile phase (acetonitrile/water (50:50v/v)+20 mM acetic acid) on the analytical column Phenomenex Luna C18 5 microm (150 mm x 4.6 mm) at room temperature. Detection was performed by a Micromass Quatro LC mass spectrometer in the reaction monitoring mode using positive electrospray ionization (ESI+). The MS-MS ion transition monitored were m/z 253>208 for OXC, m/z 255>194 for MHD and m/z 247>204 for IS. Over the range 20-5250 ng/ml for OXC and 40-10,500 ng/ml for MHD, the calibration curves were defined by the following equations: y = 0.00568 + 0.00296x -5.70e - 8x(2) and y = 0.00749 + 0.00178x - 5.70e - 8x(2) for OXC and MHD, respectively. All coefficient of determination (r(2)) were close to unity (0.9986-0.9994). The lower limits of quantification obtained as a result of the LLE procedure was 20 ng/ml for OXC and 40 ng/ml for MHD. The statistical evaluation of the developed method was conducted by examining within-batch and between-batch precision data, which were within the required limits. The suitability of the assay for pharmacokinetics studies was determined by measuring OXC and MHD concentration after administration of a single 10 ml of OXC oral suspension (6%) in plasma human of healthy volunteers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anticonvulsants / blood*
  • Anticonvulsants / chemistry
  • Anticonvulsants / metabolism*
  • Carbamazepine / analogs & derivatives*
  • Carbamazepine / blood
  • Carbamazepine / chemistry
  • Carbamazepine / metabolism
  • Chromatography, Liquid / methods*
  • Humans
  • Molecular Structure
  • Oxcarbazepine
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Time Factors

Substances

  • Anticonvulsants
  • Carbamazepine
  • Oxcarbazepine