BIURET TEST

This test is given by compounds containing two or more peptide bond (CO-NH group). Since
all proteins and peptides possessing at least two peptide linkage for example, tripeptide gives
positive biuret test. Thus, the biuret test can't be used for individual amino acids, because those
aren't connected with a peptide bond. The detection of the presence of proteins in biological
fluids is the principle used in the biuret test. Biuret Reagent contains hydrated copper sulphate
that provides the Cu (II) ions which form the chelate complex. Cu (II) ions give the reagent its
characteristic blue colour. It also contains potassium hydroxide solution that provides alkaline
medium. Other than that it also consists of potassium sodium tartrate which stabilizes the
chelate complex. Alkaline CuSO4 reacts with compounds containing two or more peptide
bonds to give a violet coloured product. In an alkaline solution copper II is able to form a
complex with the peptide bonds. Once this complex has been formed, the solution turns from
a blue colour to a purple colour. The deeper the blue-purple colour, the more peptide-copper
complexes that have been formed.
In this experiment, three samples of protein are used which are albumin, casein and peptone.
All three consist more than two peptide linkages thus gives positive results for biuret test.
Casein gives more purple coloured product and the fastest one to react followed by peptone
and albumin. After ten minutes, there are blue precipitates at the bottom of the test tube for all
three sample. Casein has the most concentrated precipitates followed by peptone and albumin.
This may be because both of the sample have been diluted as it looks runny compared to the
casein sample. It could also be that the sample has been oxidized because it has been left
without a proper storage. To overcome this, the sample should not be diluted and need to be
use right after the sample is prepared. All in all, it could be said that casein formed more
peptide-copper complexes than the rest. Nevertheless, all of them gives positive test for biuret
test.

Figure 1: The biuret complex combines copper II with the peptide

bonds, changing the color of the sample
REFERANCE
Laura Foist. What Does Biuret Test For? - Method & Equation. Retrieved from Study.com:
https://study.com/academy/lesson/what-does-biuret-test-for-method-equation.html on 10
November 2019
Gaurab Karki. (2018). Biuret test: Principle, requirements, Procedure and Result. Retrieved
from OnlineBiologyNotes: https://www.onlinebiologynotes.com/biuret-test-principle-
requirements-procedure-and-result/ on 10 November 2019