Exp2 - Anusha Vadlamudi Partner Christen Burns TA Lindsay Aldridge Chem 241 Lab Section 401 Room 300 Experiment 2 Bioanalytical Analysis of Glucose

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Anusha Vadlamudi Partner: Christen Burns TA: Lindsay Aldridge Chem 241 Lab; Section 401; Room 300 Experiment 2: Bioanalytical Analysis of Glucose in Sports Drinks (5/14/08)
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Honor Code Pledge: I pledge that no unauthorized assistance has been given or received in the completion of the work presented in this report. Introduction: The objective of the experiment is to determine the concentration of various real world drink samples using a spectrophotometric enzyme assay. The real world samples that will be used are sport drinks like Gatorade and Propel with different dilutions. In formulating the sports drinks, it’s important to find a balance of providing enough energy in a source that’s easily metabolized by the body while making sure not to overload with carbohydrates. The sports drinks, therefore, have two kinds of sugars: simple and complex. Monosaccharides, which include glucose and fructose are simple sugars and disaccharides such as sucrose are more complex sugars. For a disaccharide like sucrose to be metabolized by the body, it needs to be broken down into monosaccharide units of Glucose and Fructose. Bioanalytical sensors are devices used for the detection of an analyte that combine a biological component with a physicochemical detector component. The fundamental principle behind a biosensor is that a selective chemical receptor is associated with a transducer, which then translates the recognition into a measurable value in the form of color change, fluorescence, change in conductivity, change in mass, or a shift in electrochemical potential. The bioanalytical analysis of the experiment will use the specificity of enzymes to their respective substrates. Enzymes are biomolecules that increase the rates of chemical reactions while being unaffected by the reaction. Enzymes are usually very selective and only act upon a specific substrate and are thus meant to catalyze specific reactions.
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The experiment will use a spectrophotometric enzyme assay for detecting glucose. The assay entails the following two reactions: (1) 2 2 cos 2 cos O H ne Glucolacto D O e Glu D Oxidase e Glu + - + - - β (2) de ferricyani O H de ferrocyani O H Peroxide h Horseradis + + 2 2 2 2 (yellow color) Glucose Oxidase is an enzyme that is specific to glucose, and therefore, this coupling reaction would not occur with fructose. In reaction (1), Glucose Oxidase catalyzes the oxidation of glucose with 2 O to form -D-Glucolactone and hydrogen peroxide. The 2 2 O H from reaction (1) reacts with ferrocyanide in reaction (2), catalyzed by Horseradish Peroxide, and the ferrocyanide is oxidized to ferricyanide. The ferricyanide product has a distinct yellow color and since the wavelength of light in yellow is 420 nm, the amount of ferricyanide produced can be determined by finding the absorbance of the solution at this wavelength. The amount of ferricyanide will tell us the amount of glucose because from the reactions (1) and (2) it can be seen that a mole of
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This note was uploaded on 02/23/2010 for the course CHEM 241L taught by Professor Domenictiani during the Spring '08 term at UNC.

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Exp2 - Anusha Vadlamudi Partner Christen Burns TA Lindsay Aldridge Chem 241 Lab Section 401 Room 300 Experiment 2 Bioanalytical Analysis of Glucose

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