BCH2333 - Experiment #1 - Lab #1: Carbohydrates Lab BCH2333...

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Lab #1: Carbohydrates Lab BCH2333 Section: B2 (Tuesday morning 1 st week) TA: Ian Restall Date Preformed: January 19 th , 2010 Team #9 Riyad Hasim (5259800) Lab Partner: Meina (5211095)
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Introduction In living organisms, there are a group on compounds, carbohydrates, which play a vital role in the metabolism and functioning of that organism. The main use for carbohydrates is that they provide sources of energy to that organism. These carbohydrates are stored in organisms as polysaccharides, which is the polymerization of many sugars, like glycogen and starch. There are two main structures of carbohydrates of either heteropolymers or homopolymers. Homopolymers with unusual linkages, [ β(1→4) or α(1→6)], examples of this are cellulose and dextran respectively. Heteropolymers consists of sugars, amino sugars, uronic acids etc. Examples of heteropolymerized carbohydrates are hyaluronic acid and chondroitin. In this investigation, carbohydrates will be worked with and separated based on their molecular size. This first thing that will be done is a dialysis with a starch-glucose solution. This separation occurs because the glucose can pass through the semi-permeable membrane while the starch has a tougher time doing so. The second method of separation that will be preformed is gel filtration chromatography. This separation uses hydrated beads with a highly controlled porosity. The solution is placed in the column with an aqueous buffer. The molecules are then separated biggest to smallest. After this separation is done the solution will be separated into different fractions. Once the separations have been completed, a glucose test was performed on specific fractions of the solution. Glucose Oxidase-peroxidase will be used for this reaction. This first test will test for the presence of glucose in specific fractions, where the glucose will be proposed to be. Similarly, and iodine test will be done to test for the starch in the fractions that are expected to have starch. Experimental Procedure Refer to pages 11- 14 in ‘BCH 2333: Introduction to Biochemistry Laboratory Component’ manual. Observations Important Steps Observations Experiment #1: Dialysis When the dialysis tube filled with 10 mL of starch glucose solution was placed into the beaker filled with 200 mL of water, diffusion was occurring. Experiment #2: Gel Filtration Chromatography After 55 minutes had elapsed, the dialysis bad was taken out of the beaker, both solutions were still colorless Once the solution was loaded, 30 fractions of 90 drops were collected. Experiment #3: Glucose Oxidase Various tests were performed on the assayed solution, glucose oxidase-peroxidase and different sugar solutions including the internal and external contests from the dialysis experiment When the 2N H 2 SO 4 was added to each test tube, some of the solutions changed color while others seemed to stay colorless.
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Experiment #4: Iodine Reaction
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BCH2333 - Experiment #1 - Lab #1: Carbohydrates Lab BCH2333...

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