Abstract
Background: Dexamethasone (DEX), delivered through newly developed selfnanoemulsifying drug delivery systems, which is administered to rabbit eyes, has been presented. The instrumental parameters of the multiple LC-MS/MS methods in the literature are modified according to rabbit vitreous liquid and current laboratory conditions. Methods: Quick optimization of the chromatographic and mass spectrometric parameters was done by inspecting the available literature for the analysis of DEX by LC-MS/MS from biological matrices. Chromatographic separation was achieved in the reverse mode using C18 (50x2.1mm, 5μm) as a stationary phase and acetonitrile and water with ammonium acetate as a mobile phase in gradient elution. Quantitation was done in multiple reaction monitoring (MRM) mode by following the transition of m/z 393 > m/z 373 in positive ion mode. The method was also validated in terms of selectivity, within-day accuracy and precision, linearity, and limit of detection (LOD). The extraction of DEX from rabbit vitreous liquid samples was carried out by protein precipitation using acetonitrile: water (70:30, v/v). Results: DEX and beclomethasone (IS) were successfully separated and detected under optimized experimental settings. The method was selective for DEX and linear in the range of 0.5 and 250 ng/ml. The lower limit of quantification (LLOQ) was determined to be 0.238 ng/ml. The percent relative standard deviation (RSD) and recovery (%) of the low, medium, and high calibration levels were below 10% and within the range of 111%-114%, respectively. RSD (%) and recovery (%) of the LLOQ were below 17% and 82%, respectively. The validated method was successfully applied for the determination of pharmacokinetic properties of newly formulated dexamethasone selfnanoemulsifying drug delivery systems (DEXSNEDDS) used to administer DEX intravitreally to the rabbit. Conclusion: LC-MS/MS conditions for the analysis of LC-MS/MS were determined by examining relevant literature for accomplishing simple and practical optimization of the experimental parameters, followed by method validation and analysis of rabbit vitreous liqiud. As conclusion, pharmacokinetic data of DEXSNEDDS has been obtained in the most accurate, sensitive, economical and rational way possible.
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