Development and Validation of an Ultra-Performance Liquid Chromatographic-Tandem Mass Spectrometry (Uplc-Ms/Ms) Method for Determination of Betamethasone or Dexamethasone in Pharmaceutical Preparations

Abstract

Development and Validation of an Ultra-Performance Liquid Chromatographic-Tandem Mass Spectrometry (UPLC-MS/MS) Method for Determination of Betamethasone or Dexamethasone in Pharmaceutical Preparations, Hacettepe University, Institute of Health Sciences, Program of Analytical Chemistry, PhD Thesis, Anakara, 2013, Betamethasone (BTM) and Dexamethasone (DXM) are highly potent fluorinated synthetic glucocorticosteroids. They are widely used for the treatment of inflammation, allergies and adrenaral cortex insufficiency. Chromatographic methods for determination of BTM or DXM in pharmaceutical preparations (tablets, injectable ampoules and eye/ear drops) by Ultra-Performance Liquid Chromatography-Tandem Mass Spectroscopy (UPLC-MS/MS) using negative Electrospray Ionization (- ESI) at MRM mode were developed. The chromatographic analysis were performed on UPLC Acquity BEH C18(50 mm × 2.1mm, 1.7 µm) column was eluted using stepwise gradient with mobile phases of acetonitril and 0.1% (v/v) acetic acid in water. Detection of DXM and BTM was performed by triple quadrupole tandem MS using negative ESI at 451 > 361 m/z transition on MRM mode. Mefruside was used as an internal standard and traced at 381 > 189 m/z transition on MRM mode. Developed methods were validated according to analytical method validation guideline from ICH. The methods were linear over the concentration range of 10 to 1500 ng mL?1 for both BTM and DXM. Limits of detection (LOD) for BTM or DXM were1ng mL?1 while limit of quantitation (LOQ) was 5 ng mL?1 for BTM or DXM. The intra- and inter-accuracy values expressed as relative error (RE %) were ?1.08 -1.76 and -0.93 ? 1.98 for BTM and DXM, respectively. On the other hand, the intra- and inter-day precision expressed as relative standard deviation (RSD %) were 0.28-1.80 and 0.23-1.02 for BTM and DXM, respectively. The standard solutions of BTM and DXM were stable at + 4 ºC for 6 months. The method was also rapid, specific, rugged and robust. Validated methods were also successfully applied for the analysis of some commercially available pharmaceutical preparations of BTM (Celestone® Tablet and Celestone® Injectable Ampoule) and of DXM (Dekort® tablet, Dekort® injectable ampoule and Onadron ® Simple eye/ear drop). Therefore, this method is suitable for determination of BTM or DXM in their pharmaceutical preparations.