College of Health Sciences Education

3rd Floor, DPT Building

Matina Campus, Davao City
Telefax: (082)
Phone No.: (082)300-5456/300-0647 Local 117

PROGRAM:BACHELOR SCIENCE OF NURSING

CN112/L BIOCHEMISTRY
Laboratory Exercise No.6
BIURET TEST FOR THE PRESENCE PROTEINS
https://youtu.be/pgSKISrnxxU

INTRODUCTION
Biurest Test for the Presence of Proteins

The reagent used in the Biurest Test is a solution of copper sulfate (CuSO4) and potassium
hydroxide (KOH). The KOH is there to raise the pH of the solution to alkaline levels; the
crucial components in the copper ion from the CuSO4.

When the peptide bonds are present in this alkaline solution, the copper ions will form a
coordination complex with four nitrogen atoms involved in peptides bond.

Copper Sulfate Solution is a blue color, but when the Copper Ions are coordinate with the
nitrogen atoms of these peptide bonds, the color of the solution change form blue to violet.
This color change is dependent on the number of peptide bonds in the solution, so the more
protein, the more intense the change. When the peptides are very short, the solution turns
pink in color rather than violet.

OBJECTIVES
At the end of the experiment, each student will be able to:

1. Test presence of proteins with the use of biuret.

2. Know the procedure and perform the protein sample preparation.

MATERIALS NEEDED
1-200ml beaker, stirring rod, 3-test tubes, test tube rack, test tube holder, test tube brush, 2
medicine dropper, 50ml distilled water, 2ml KOH solution, 2mlCuSO4 solution, 2- egg (raw).

PROCEDURE
Protein Sample Preparation
1. Separate the egg albumin and the egg yolk: In this experiment, you will use onlythe
albumin (egg white).
2. Place the albumin in the beaker and mix with 10-20 ml of distilled water. Youmay add
more water if the solution is still sticky.
3. Shake the contents thoroughly but try to avoid creating bubbles and leave it forthe next
procedure.
 College of Health Sciences Education
3rd Floor, DPT Building
Matina Campus, Davao City
Telefax: (082)
Phone No.: (082)300-5456/300-0647 Local 117
The Biuret Test
1. Fill a test tube one-third of the solution to be tested (albumin solution). Prepareanother
tubes as your control with distilled water only.
2. Add 10 drops of KOH to each tube and shake them thoroughly.3. Add copper sulfate
solution one drop at the time. Shake the solution after eachdrop. This is very important to
avoid coagulation.4. Keep adding the copper sulfate solution until you will notice color
change,usually after 5-9 drops

EXPECTED RESULT
Since this complex absorbs light in 540 nm, it appears violet. In the presence of protein, it
changes its colour from blue to violet.
Solution turns violet from blue
Solution turns pink from blue
MEMBERS:
CENA RITCHELL ANO
CANADA LIAM EMMANUEL BACUS
CAMANSI CRIA JIMENES
CASTRO NORTRO NORLYN TAHA
CASTRO SHARA TAHA
CATARROJA BENJIE CARICUNGAN
DEALA NATHALIE LABASTIDA
FELICILDA ALYSSA KATE CASONETE
HERMOCILLA MARICAR ABAR