Biuret Test Essay

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Biuret Test The Biuret Reagent is made of sodium hydroxide and copper sulfate. The blue reagent turns violet in the presence of proteins, and changes to pink when combined with short-chain polypeptides. http://www.biosci.ohiou.edu/introbioslab/Bios170/170_2/biuret.htm Biuret test is used for detecting the presence of peptide bonds. It relies on the reduction ofcopper(II) ions to copper(I), the latter form a complex with the nitrogens of the peptide bonds in an alkaline solution. A violet color indicates the presence of proteins The Biuret reaction can be used to assess the concentration of proteins because peptide bonds occur with the same frequency per amino acid in the peptide. The intensity of the color, and hence the absorption at 540 nm, is directly proportional to the protein concentration, according to the Beer-Lambert law. http://en.wikipedia.org/wiki/Biuret_test BENEDICT’S REDUCING SUGAR TEST The Benedict's’s Reducing Sugar Test will test positive on both monosaccharides and some disaccharides. It will test positive for all monosaccharides and disaccharides that can donate electrons to a solution of copper ions. For example, fructose, glucose, maltose and lactose. The procedure involves adding an equal amount of Benedict's’s solution to the test solution in a test tube. Then the test tube is heated to speed the chemical reaction. Electrons on the monosaccharide or disaccharide are removed by the ionic copper in the Benedict's’s solution. This results in copper precipitating out of the solution. See the picture below for a positive test and a negative test. The limitation of the test is that we cannot tell if the solution is a mono or disaccharide. Unless we pair this test with Barfoed’s Test. If we run the Barfoed’s Test first and get a negative result and then run the Benedict’s Reducing Sugar Test second and get a positive test; we know the

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