Abstract

STUDY OF FACTORS AFFECTING ON SOLVENT EFFECT AND PEAK SHAPE IN REVERSED PHASE CHROMATOGRAPHY

Xiaobing Zhan and Dr. Vishal Shinde*

Abstract

In the application of chromatography, the Peak-shape problem is the most common trouble. It causes integration problems and poor resolutions. The goal of the present work is to, study the factors affecting on solvent effect and the peak shape of the chromatogram such as of different pH in the diluent (pH 3.0- pH6.8), addition of 0.5% SDS the solution, Different column temperature (300C-500C), Different column type (Column 1: 100-5-C18 (250×4.6mm, 5μm), Column 2: Zorbax SB-C18 (150×4.6mm,5μm) and mobile phase with different composition, In this study Aniline used as analyte and set the Chromatographic conditions as Column: 100-5-C18 (250×4.6mm, 5μm) Detection wavelength: 254mn, Flow rate: 1ml/min, Injection volume: 10μl. It explained the reason for the solvent effect, and it redefined the concept of the solvent effect. The solvent effect caused due to the above difference is because of the large difference between the distribution coefficient of the stationary phase-diluent and the distribution coefficient of the stationary phase-mobile phase. It results from the three-media effect i.e. stationary phase, diluent and mobile phase. Therefore, when assessing the risk of solvent effects, it may combine with the specific situation of these three media and to assess the strength of the interaction between the three media. In the distribution coefficient of the analyte in the stationary phase-diluent and the stationary phase-mobile phase, the partition coefficient may affect the different pH in the diluent, addition of SDS solution in the diluent, Different column temperature, Different column type and different mobile phase with different compositions and it can also be adjusted to optimize the effect of the solvent effect.

Keywords: Liquid chromatography, Solvent effect, Reversed-Phase chromatography, Peak shape, Distribution coefficient, Aniline.