Lab No.

1
(Insert catchy title)

Protein Denaturation and Protein Test

I. Objective
1. To examine how various environmental factors (temperature, pH, , and
chemical agents) induce denaturation, providing insights into protein stability
and functionality under different conditions.
2. To understand how denaturation affects the catalytic activity of enzymes.
3. To characterize how different proteins respond to stress conditions, enhancing
our understanding of protein resilience and adaptability.
II. Materials

● Alcohol ● Test Tubes

● Water ● Test tube rack
● Vinegar ● Test tube brush
● Egg ● Alcohol lamp
● Hot plate ● Spatula
● Wash bottle ● Beaker
● pipette
III. Procedure
1. Pour droplets of rubbing alcohol into a glass test tube, room temperature
water into another, and the rest of the water into test tubes.
2. Crack egg into the bowl, removing the yolk.
3. Cut the egg white into pieces so you can add ¼ into each test tube.
4. Heat up the water for your hot water treatment and pour into one of the empty
test tubes.
5. Quickly put droplets of egg yolk into the boiling water. Then put droplets into
the alcohol, into the room temperature water, and the remainder into the final,
empty test tube.
6. Observe any immediate changes that occur in terms of egg white color and
consistency.
7. Wait for 30 minutes.
8. Inspect the state of the egg whites in each treatment and note how they may
have changed over time.
IV. Data and Observation
A. Protein Denaturation
Sample Observation

1. Egg white + Vinegar When the egg white was added to the
vinegar, they did not mix together
initially. The egg white remained mostly
intact.
 Over time (30 minutes), small bubbles
began forming at the surface where the
egg white and vinegar met.

2. Egg white + Rubbing alcohol When the egg white was added to the
rubbing alcohol, it quickly coagulated
and turned cloudy upon stirring.

After 30 minutes, the egg white

appeared fully cooked, firm, and
solidified, indicating protein
denaturation caused by the alcohol.

3. Egg white + Boiling water When the egg white was added to the
boiling water, it started to cook almost
immediately, turning a bit white and
opaque. It didn’t fully solidify, but you
could see it was starting to coagulate,
giving it a slightly cooked appearance

4. Egg white + Tap water When the egg white was added to the
room temperature water, it did not mix
and remained separated.

After 30 minutes, small bubbles formed

at the surface, but the egg white itself
showed no significant change in color.

V. Conclusion/Generalization
 Protein Test
I. Objective
1. To determine the concentration of proteins in various samples (e.g.,
tofu, egg whites, Milo energy drink) using methods such as the Biuret
test, Millon's test, Ninhydrin test, and Xanthoproteic test.
2. To assess how proteins behave under various conditions, including
temperature changes, and the presence of denaturing agents.
II. Material
● Tofu ● Test tube rack
● Water ● Test tube brush
● Egg ● Alcohol lamp
● Milo ● Spatula
● Hot plate ● Beaker
● Wash bottle ● pipette
● Test Tubes

III. Procedure
A. Xanthoproteic Test
1. In three test tubes, fill in a small amount of egg white, Milo,
and tofu.
2. Add 5 drops of sulfuric acid on each test tube.
3. Shake the test tubes.
4. Heat the test tubes in a water bath.
5. Check for changes.

B. Biuret Test

1. Fill in three test tubes with a small amount of sliced tofu, egg
white, and Milo.
2. Add 2 ml of NaHCO3
3. Add 5 to 6 drops of copper sulfate.
4. Shake to mix the ingredients well.
5. Observe.

C. Ninhydrin Test
1. Fill in the three tubes with Milo, tofu, and egg white.
2. Add 1 ml of Ninhydrin solution.
3. Boil the mixture.
4. Observe.
D. Millon's Test
1. Fill in three test tubes with Milo, tofu, and egg white.
2. Add three drops of Millon's reagent and shake well.
3. Observe changes.
IV. Data and Observation
 A. Xanthoproteic Test

Sample Observation

White Egg + Sulfuric Acid When sulfuric acid was added to the white
egg, it coagulated immediately upon
contact. After heating in the water bath,
the mixture formed a singular clump,
resembling a poached texture.

Milo + Sulfuric Acid Before heating, the Milo and sulfuric acid
mixture was white and cloudy with some
impurities. After boiling, the mixture
separated, forming tiny clumps that
floated, and the color faded significantly.

Tofu + Sulfuric Acid Before heating, the tofu turned brown

when sulfuric acid was added. After
boiling, the tofu melted and became
darker in color.

B. Biuret Test

Sample
Observation

Tofu + Copper Sulfate When copper sulfate was added to the tofu,
smaller pieces of tofu settled at the bottom,
while larger pieces floated to the surface.
No other significant changes were
observed.

White egg + Copper Sulfate When copper sulfate was added to the
white egg, the mixture turned
yellowish-green. The chemicals did not
mix well, and the egg white settled at the
bottom of the test tube. White impurities
appeared on the surface, and bubbles
formed during the reaction.

Milo + Copper Sulfate When copper sulfate was added to the

Milo, the mixture turned a dark grayish
color. No other changes were observed.

C. Ninhydrin Test
 Sample Observation

Egg white + Ninhydrin

solution After adding Ninhydrin solution to the egg
white and boiling, the solution turned
purple. The egg white cooked underneath,
and the purple color seeped into about half
of the egg white.

Tofu + Ninhydrin solution After adding 1 ml of Ninhydrin solution to

the tofu and boiling, the mixture turned
violet.

Milo + Ninhydrin solution After adding 1 ml of Ninhydrin solution to

the Milo and boiling, the mixture turned
deep purple.

D. Millon’s Test

Sample Observation

Egg white + Millon’s

reagent After adding three drops of Millon’s
reagent to the egg white and shaking, the
mixture became sticky with bubbles
forming at the top. Tiny white particles or
white impurities were also observed.

Milo + Millon’s reagent

After adding three drops of Millon’s
reagent to the Milo and shaking, foam
formed at the top of the mixture.

Tofu + Millon’s reagent

After adding three drops of Millon’s
reagent to the tofu and shaking, the reagent
was absorbed by the tofu, with no change
observed.

V. Conclusion/Generalization