CHM 3103 Lab Report # 3.

GOKHUL, Raja.

October 14, 2014.

Registration #: 14/0705/0763.

Group: A Leanna Simon, Alicia Sukhdeo, Julian Monize, Amanda O Neil and Latoya Douglas.
Title:

Investigation of some Physicochemical Properties of Food Proteins.

Objectives:

1. To Demonstrate Denaturation and to Determine the Coagulation Point of Egg

Albumin
2. To Investigate the Gelation of Knox ORIGINAL Gelatine (Unflavored).

Principle:

1. Demonstration of Denaturation and Determination of the Coagulation Point of

Egg Albumin
The disruption and or breaking of the tertiary and the secondary structures
of proteins occur during the denaturation of protein. The primary structure
remains intact because denaturation reactions are too weak sever the peptide
bonds. Basically, denaturation produces a random shape from the normal alphahelix and beta sheets in a protein by uncoiling them. The secondary and the
tertiary structures of proteins are held in shape via the following interactions;
hydrogen bonding, salt bridges, disulfide bonds, and non-polar hydrophobic
interactions. These interactions are affected by various parameters, which are
ultimately responsible for denaturation; heat, alcohol, acids and bases, heavy
metal slats and reducing agents etc.
Nonpolar hydrophobic interactions along with hydrogen bonds are
disrupted via the application of heat. As the heat applied increases, the kinetic
energy increases, which causes the vibration of the molecules to become so
violent, that the disruption of the bonds occurs.
The salt bridges which are held in place via ionic charges can be disrupted
via acids and bases. A type of double replacement reaction transpires where the
positive and negative ions in the salt change partners with the positive and
negative ions in the new acid or base added. The salt bridge has the effect of
straightening an alpha helix so when it is disrupted, the secondary structure is
affected. The coagulation of protein is the most common observation in the
denaturation process.
Coagulation is described as the phenomenon in which a liquid thickens
into a semi-solid form. When Egg White coagulates, it transforms from a clear,
transparent mass into a mass that is white and opaque. The coagulation of Egg
White occurs around the temperature of 60 oC 65 oC. A few factors which affect
the coagulation of Egg Protein are; temperature, time, concentration of protein,

Page 1 of 5

CHM 3103 Lab Report # 3.1

GOKHUL, Raja.

October 14, 2014.

Registration #: 14/0705/0763.

Group: A Leanna Simon, Alicia Sukhdeo, Julian Monize, Amanda O Neil and Latoya Douglas.
salt content and its concentration, reaction of the egg solution or mixture, and
sugar.
As the temperature raises the rate of coagulation rises and at very high
temperatures, coagulation occurs very rapidly. The amount of coagulant produced
at a specific temperature varies directly to the amount of time the sample is
subjected to that specific temperature. As pH decreases (acidity increases),
coagulation of Egg White increases. A more alkaline Egg White will undergo less
coagulation than one at a neutral pH. The optimum pH for Egg White coagulation
is 4.7. The temperature at which the coagulation of Egg White occurs (60 oC 65
o
C) increases when the egg white is diluted.
2. Gelation of Gelatin.
Gelatin is a protein substance. It is obtained via boiling animal bones and
connective tissue containing collagen in water or dilute acid. In its purified state, gelatin
is colorless, transparent, brittle, odorless, and tasteless. Gelatin dissolves in hot water and
forms a gel or jelly upon cooling. When placed in cold water; gelatin takes up five to ten
times its own weight and swells to an elastic, transparent mass.
As the pH decreases or acidity increases the strength of the gel decreases and the
time taken for the gel to settle also increases. Only large quantities of sugar would hinder
the gelation of gelatin in a significant manner. Cooling increases the rate of gelation and
the firmness of the gel.
In this experiment, the gel strength of various mixtures of gelatin and other
commodities will be assessed via the following the relationships;
1.

sag=

2.

sag

H 1H 2
100
H1

1
Strenght of gel

Procedure:

Same as Handout.

Results:

The results gathered from lab # 3.1 are tabulated as follows:

1. Demonstration of Denaturation and Determination of the Coagulation Point
of Egg Albumin
Page 2 of 5

CHM 3103 Lab Report # 3.1

GOKHUL, Raja.

October 14, 2014.

Registration #: 14/0705/0763.

Group: A Leanna Simon, Alicia Sukhdeo, Julian Monize, Amanda O Neil and Latoya Douglas.

The results obtained from Demonstration of Denaturation and Denaturation of

the Coagulation Point of Egg Albumin experiment are tabulated in table 1 on
the following page:
Table 1: Results obtained from the Demonstration of Denaturation and
Denaturation of the Coagulation Point of Egg Albumin experiment
Protein
Egg Albumin (from test
tube #2 with the buffer
soln. pH 4.7)

Coagulation Temperature/C
78

2. Gelation of Gelatin
The results obtained from Gelation of Gelatin experiment are tabulated in
table 2 below:
Table 2: Results obtained from the Gelation of Gelatin experiment.
Percentage

Gel Strength (Inversely

Sag / %
0

proportional to % sag)
Extremely Strong

After 1 day

Mixtures
Gelatin + Water
Gelatin + Citric Acid
Gelatin + Cane Sugar

2.22

Extremely Strong

Extremely Strong

Extremely Strong

Maize + Citric Acid + Cane Sugar

Discussion:

1. Demonstration of Denaturation and Determination of the Coagulation Point of

Egg Albumin
There are numerous means by which Proteins can be denatured. Some of
the means utilized in this experiment included; heating, mechanical action
(beating/whisking) and the addition of an acid or a base. All of the systems
eventually experienced coagulation. Therefore it can be inferred that the means
utilized were successful for denaturing protein since denaturation must precede
coagulation. Furthermore this inference is quite reasonable since the means in
question do denature protein as addressed in the principle.
Page 3 of 5

CHM 3103 Lab Report # 3.1

GOKHUL, Raja.

October 14, 2014.

Registration #: 14/0705/0763.

Group: A Leanna Simon, Alicia Sukhdeo, Julian Monize, Amanda O Neil and Latoya Douglas.
The system which had the buffer solution experienced coagulation at
78C. This is above the literature range of 60 oC 65 oC. The reason why
coagulation occurred at a higher temperature are because, the Egg White was
diluted to 5%. Dilution increases the coagulation temperature.
The acidic and basic systems, however, did not coagulate with system
containing the buffer solution (pH: 4.7) at 78 C. Therefore it can be inferred that
1. The addition of strong acids (HCL) and strong bases (KOH) affects the
coagulation of egg albumin. 2. A pH of 4.7 is ideal for the coagulation of egg
albumin, since when the buffer solution of pH 4.7 was added to the acidic and
basic systems and re-heated, coagulation of the egg albumin occurred.
2

Conclusion:

Gelation of Gelatin.
All four of the gels prepared were extremely strong as all of their % sags
were extremely low. As mentioned in the principle Gelatin + water form a
strong gel. All of the gels had 0% sag barring the gel that contained Gelatin
and Citric Acid, which had a % sag of 2.22. The reason behind this may be the
addition of acid, since the addition of acid does decrease the strength of a gel.
However citric acid is a weak acid, hence its effect is not very profound. The
amount of sugar added to the gelatin and sugar gel was not substantial to
affect the gel strength, since only the addition of large amounts of sugar would
decrease gel strength. As stated already, only large quantities of sugar and
considerably strong acids would decrease the gel strength, and since a small
quantity of sugar and weak acid was used, this is why the gel strength of the
Gelatin + Citric acid + sugar gel was not affected.
Since all of these gels were cooled, the gelation process was aided further.
It can be concluded that;
1. (a) Egg White can be denatured via several means, such as, heat &
variation of the pH of the system. (b) Dilute Egg White solution (5%)
coagulates at 78 C at a pH of 4.7. (c) The pH of the system does affect
the coagulation/ coagulation point of Egg White.
2. Knox ORIGINAL Gelatine (Unflavored) forms very strong gels with

Water.
Citric acid (weak acid).
Small amounts of sugar and
Citric acid (weak acid) + Small amounts of sugar.
Page 4 of 5

CHM 3103 Lab Report # 3.1

GOKHUL, Raja.

October 14, 2014.

Registration #: 14/0705/0763.

Group: A Leanna Simon, Alicia Sukhdeo, Julian Monize, Amanda O Neil and Latoya Douglas.

References:
1. Ophardt, C. (2003). Denaturation Protein. Retrieved November 5, 2014, from
2

http://www.elmhurst.edu/~chm/vchembook/568denaturation.html
"Gelatin." Microsoft Encarta 2009 [DVD]. Redmond, WA: Microsoft Corporation,

2008.
3. Vaclavik, V., & Christian, E. (2013). Proteins in Food. In Essentials of Food Science (4th
ed., p. 185). Springer Science & Business Media.
4. Lowe, B. (1937). Gelation And Stiffening Power Of Gelatin. In Experimental Cookery
From The Chemical And Physical Standpoint Read more:
Http://chestofbooks.com/food/science/Experimental-Cookery/index.html#.VFo4RTTF54#ixzz3ICoU1tef. John Wiley & Sons.
5. Srilakshmi, B. (2003). Eggs. In Food Science (3rd ed., p. 135). New Delhi: New Age
International.

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