EXPERIMENT - TEST FOR PROTEINS USING BIURET REAGENT

OBJECTIVE The purpose of this lab was to determining the presence of protein using Biuret Reagent.

PROBLEM STATEMENT How to determine the presence of protein using Biuret Reagent?

HYPOTHESIS Presence of protein can be detected by using Biuret Reagent.

VARIABLES:Manipulated- Type of solution Responding: Presence of protein Fixed: Volume of solution

APPARATUS Test tubes in test tube rack Stock bottles of: Biuret Reagent Distilled water (dH2O) Starch solution Egg albumin

 Glucose solution Vortex mixer

PROCEDURE 1) The correct solution was pipet 1ml into the test tubes as described in table. 2) 2 ml of Biuret reagent was added to each test tube and the mixture was agitate by shaking the test tubes from side to side or with a vortex mixture. 3) Wait for 2 minutes and then the colour after reaction of the mixture was recorded in table.

RESAULT TUBE CONTENTS COLOUR BEFORE REACTION 1) 2) 3) 4) dH2O Starch Egg albumin Glucose Colourless Colourless Yellowish Colourless COLOUR AFTER REACTION Blue Blue Purple Blue Absent of protein Absent of protein Present of protein Absent of protein CONCLUSION

DISCUSSION Proteins are the most complex and functionally diverse molecules of living organisms.Proteins are created by RNA during DNA Transcription and Translation. The base elements of proteins are C, H, O and N. The monomers of proteins are 20 different amino acids. The amino acids are bonded together in unique combinations to create a polypeptide chain, the protein polymer. In this lab will test for the presence of protein using the Biuret Test. Like the Benedicts Reagent, Biuret Reagent contains copper ions. These copper ions reflect off closely clustered amide groups of proteins casting a violet color to a solution with proteins. This violet color is a positive reaction in a Biuret Test. TEST POSITIVE OBSERVATION The solution turns from blue to violet( purple) The solution turns from blue to pink NEGATIVE No change /The solution remains blue INFERENCE Proteins are present Peptides are present ( Peptides or peptones are short chains of amino acid residues) No protein is present

SOURCE OF ERROR 1) Do not put same volume of stock solution. 2) The mixture does not shaking well. 3) The time for record the colour after reaction does not accurate 2 minutes.

LIMITATION 1) Insufficient of time to do this experiment. 2) Inside pippet have air bubble, so that the amount of solution does not same.

CONCLUSION Presence of protein can be detected by using Biuret Reagent. Hypothesis was accepted.

FURTHER WORK 1) Longer the time to do this experiment to 3-4 hours. So, that I can get accurate result. 2) Increase the amount of solution. 3) Use other stock of solution.

REFERENCE Internet http://biology.unm.edu/ccouncil/Biology_124/Summaries/Macromol.html http://www.brilliantbiologystudent.com/biuret_test.html