HPLC Lab

HPLC Lab - High Performance Liquid Chromatography (HPLC) of...

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Unformatted text preview: High Performance Liquid Chromatography (HPLC) of Caffeinated Beverages John Goodson Partner(s) : Jessica Highsmith Date Performed: 15 April 2010 Abstract: The purpose of this study was to perform an analysis of caffeine content of 5-hour Energy and Redbull energy drinks using high performance liquid chromatography with a UV-Vis detector. Quantification of caffeine will be based on a calibration curve prepared in a concentration range of 0.025 to 0.2 mg/mL. I ntroduction: High performance liquid chromatography (HPLC) is a type of column chromatography that is often used in biochemistry and analytical chemistry to separate, identify, and quantify compounds. HPLC makes use of a column that holds a packing material, referred to as the stationary phase, a pump that moves the mobile phase or phases, which is liquid in HPLC, through the column, a detector that shows the retention times of the molecules and a data processor. Retention time varies depending on the interactions between the stationary phase, the molecules being analyzed, and the solvent or solvents used as the mobile phase. Using HPLC components can be separated by distribution between two phases, one of which is stationary, in this case the packing material, while the other moves, called the mobile phase. Separation of a mixture dissolved in the mobile phase solvent allows the analyte to be measured from other molecules in the mixture and allows it to be isolated. In this case, caffeine from the energy drinks is separated from the original mixture by dissolution into the mobile solvent and passed through a column containing a packing material. Differing affinities of the polarity of the mobile and the stationary phase allow differing components to elute through the column at differing speeds. A detector allows for detection of the components as they elute out of the column. This will allow determination of the quantities of analyte in the mixture. Objective: To perform a quantitative analysis of the caffeine composition of a prepared sample of energy drinks using HPLC as a means of separating the molecular components of the sample. Materials and Methods : Apparatus: Shimadzu High Performance Liquid Chromatograph UV-Vis detector Analytical Balance Volumetric Flask Separatory Funnel Graduated Cylinder 250-mL Round Bottom Flask Vortex Bchi Rotavapor R-210 Graduated Pipette Procedure: Caffeine Standard Preparation: 1. 1 mg/mL Caffeine Stock Solution: 100 mg of caffeine were weighed on an analytical balance, quantitatively transferred to a 100-mL...
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HPLC Lab - High Performance Liquid Chromatography (HPLC) of...

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