Optimization and validation of a high performance liquid chromatography method for rapid determination of sinafloxacin, a novel fluoroquinolone in rat plasma using a fused-core C(18)-silica column.
ABSTRACT A novel, simple and rapid high performance liquid chromatographic method has been developed and validated for the determination of sinafloxacin, a new fluoroquinolone, in rat plasma using 96-well protein precipitation, fused-core C(18)-silica column (4.6mmx50mm, 2.7microm) packed with a new solid support, which is made of 2.7microm particles that consist of a 1.7microm solid core covered with a 0.5microm thick shell of porous silica.The chromatographic separation was achieved with a mobile phase of 20:80 (v/v) of acetonitrile and phosphate buffer (pH=3.0) at a flow rate of 1mlmin(-1). Fluorescence detection was employed with lambda(ex) 295nm and lambda(em) 505nm. Lomefloxacin was used as internal standard (IS). The total analysis time was as short as 3min. The method was sensitive with a limit of detection (LOD) of 2ngml(-1), with good linearity (R(2)=0.9996) over the linear range of 5-500ngml(-1). The intra-day and inter-day precision was less than 5.8% and accuracy ranged from 100.3% to 103.5% for quality control (QC) samples at three concentrations of 10, 50 and 400ngml(-1).The fused-core C(18)-silica column method offered high sample throughput, low injection volume and low consumption of organic solvents. The method was successfully employed in the pharmacokinetic study of sinafloxacin formulation product after tail vein injection to healthy rats.
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ABSTRACT: As part of the method development, the injection volume as a critical quality attribute in fast fused-core chromatography was evaluated. Spilanthol, a pharmaceutically interesting N-alkylamide currently under investigation in our laboratory, was chosen as the model compound. Spilanthol was dissolved in both PBS and MeOH/H2O (70/30, v/v) and subsequently analyzed using a fused-core system hereby selecting five chromatographic characteristics (retention time, area, height, theoretical plates and symmetry factor) as responses. We demonstrated that the injection volume significantly influenced both the qualitative and quantitative performance of fused-core chromatography, a phenomenon which is confounded with the nature of the used sample solvent. From 2 μL up to 100 μL injection volume with PBS as solvent, the symmetry factor decreased favorably by 20%. Moreover, the theoretical plates and the quantitative parameters (area and height) increased up to 30%. On the contrary, in this injection volume range, the theoretical plates for the methanol-based samples decreased by more than 60%, while the symmetry factor increased and the height decreased, both by 30%. The injection volume is thus a critical and often overlooked parameter in fused-core method description and validation.Journal of Pharmaceutical Analysis. 10/2013; 3(5):330–334.
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ABSTRACT: The need for pharmaceutical analytical laboratories to increase sample throughput and speed of analysis to meet accelerated drug development timelines has led to considerable interest in the development of fast liquid chromatographic technology. Fast liquid chromatographic pharmaceutical analysis is accomplished by developments in ultra-fast high performance liquid chromatographic instrumentation, emerging technologies in column stationary phases and high temperature liquid chromatographic analysis. This review provides an overview and gives recent developments and applications in the different strategies used in fast liquid chromatographic pharmaceutical analysis.Journal of Liquid Chromatography & Related Technologies 01/2011; 34(13):1133-1156. · 0.57 Impact Factor
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ABSTRACT: The present work describes the systematic development of a robust, precise and rapid reversed-phase liquid chromatography method for the simultaneous determination of eprosartan mesylate and its six impurities using Quality by Design principles. The method was developed in two phases; screening and optimization. During screening phase, the most suitable stationary phase, organic modifier and pH were identified. The optimization was performed for secondary influential parameters; column temperature, gradient time and flow rate using eight experiments to examine multifactorial effects of parameters on the critical resolution and generated design space representing the robust region. A verification experiment was performed within working design space and model was found to be accurate. This study also describes other operating features of column packed with superficially porous particles that allows very fast separations at pressures available in most liquid chromatography instruments. Successful chromatographic separation was achieved in less than 7 min using a fused-core C18 (100 mm × 2.1 mm, 2.6 μm) column with linear gradient elution of 10 mM ammonium formate (pH 3.0) and acetonitrile as the mobile phase. The method was validated for specificity, linearity, accuracy, precision and robustness in compliance to the ICH Q2 (R1) guidelines. The impurities were identified by liquid chromatography with mass spectrometry. This article is protected by copyright. All rights reserved.Journal of Separation Science 06/2014; · 2.59 Impact Factor