DEPARTMENT OF PURE AND APPLIED CHEMISTRY

COLLEGE OF ARTS AND SCIENCES

VISAYAS STATE UNIVERSITY

Name: Vincent Kim E. Vergara Date Performed: June 19, 2019

Lab Schedule: 7:00 – 10:00 (M-F) Date Submitted: July 1, 2019
Group No: 5 Score:

Experiment No. 11
DIGESTION OF FOODSTUFFS
ABSTRACT:
This experiment focuses on the identification of digestion as a process of hydrolysis and determination
of the hydrolysis products of carbohydrate, fat and protein digestion. Digestion is an essential process in living
organisms. It is in a form of hydrolysis process that breaks down large complex biomolecules into simpler
substances that can be easily absorbed by the intestines. The hydrolysis products of carbohydrate, fat, and
protein digestion are glucose molecules, glycerol and fatty acids, and amino acids, respectively. Each product
has its own metabolism which also has different function such that carbohydrates are essential for energy
storage; fats are for neural transmissions, and; proteins are diverse that its functions counterpart almost all in
the systems of the body.

INTRODUCTION:
Digestion is the activity of food converted into substances that can be absorbed and integrated in the
body in the process of hydrolysis by enzymes. Enzymes are essential to make the hydrolytic reactions more
effective, by lowering the activation energy of the reaction.
The first part of the experiment tackles about the digestion of carbohydrates specifically starch. The
starch solution was divided into two; one served as control the other was added with salivary amylase. Iodine
test was also used to indicate the presence of starch, which gives a blue-black color. Another test to confirm
the complete hydrolysis of starch is the Benedict’s test. The positive result for this is the presence of orange
color in the solution, which indicates that glucose, the monosaccharide in starch, is present. The second part of
the experiment is the digestion fats. This part was divided into two sections. The first one was aided with Bile
salts. The other was the digestion of fats in milk through the use of lipase (another enzyme vital to the
digestion of fat). Lipase hydrolyses the fat in milk to fatty acids which react with sodium hydroxide to lower the
pH of the mixture. This pH change is perceived by using phenolphthalein. The last part of the experiment is the
protein synthesis. Three set-ups were prepared: the first was water and HCl, the second was pepsin and HCl,
and the third was pepsin and water. The protein to be assessed was albumin, the white portion of the hard-
boiled egg.
METHODOLOGY:

A. DIGESTION OF CARBOHYDRATES
A-1. Hydrolysis if Starch
- 1 mL of saliva collected in a small beaker. Two test tubes prepared with 5 mL 1% starch. 10 drops
of saliva containing salivary enzyme added to one of the test tubes and mixed thoroughly. A spot
plate or wax paper were then prepared with 1 drop of iodine reagent for each test.
- Every two minutes, place a drop of each mixture in the spot plate or on a drop of iodine on the wax
paper. A deep-blue color indicates the starch is present. Testing the mixtures until deep-blue color
for starch no longer forms continued. Time recorded.
 A-2. Formation of Glucose
- In the test for the presence of glucose as a final product of starch digestion, 5 mL OF Benedicts
reagent to each of the test tubes and contents added. Test tubes placed on a boiling water bah for
about 5 min. Colors that form recorded. Determine if glucose present or not.

A. DIGESTION OF FATS
B1. Bile Salt
- 2 mL safflower placed in each test tubes. 8 mL of water added to one test tube. To other sample, 5
ml water and 3 mL bile salts added and were mixed thoroughly. Let the test tube stand for 30 min.
Observation recorded.
B-2. Hydrolysis by lipases
- 50 mL of milk placed in an Erlenmeyer flask. The flask was placed in a 37˚C water bath using a 400
mL beaker. Set up buret containing 0.1M NaOH. 10 mL 2% pancreatin added to the flask and
mixed thoroughly.
- 10 mL of mL mixture carefully poured into another flask. Return the rest of the fmilk to the 37˚C
water bath. PH of the mil sample was tested using a pH meter.
- 3-5 drops of phenolphthalein added to the milk sample. 0.1N NaOH from the buret titrated until a
permanent light pink color is obtained. This marks the endpoint. Number of millimeters of NaOH
required to reach the endpoint recorded.
- 10 mL of samples of the milk and lipase mixture remover for every 20 minutes until 60 minutes have
elapsed. The pH of each sample were immediately determine and then titrated with 0.1N NaOH.
Number of millimeters of NaOH used each time recorded.

B. Protein Digestion
Obtain three small pieces of the white part of a hard- biled egg. Place the pieces in three separate
test tubes. Three test tubes prepared as follows

Test tube Solutions

1 5 mL water + 1 mL 0.1N Hcl

2 5 mL pepsin + 1 mL 0.1 HCl
3 5 mL pepsin + 1 mL water

The three test tubes were placed in a water bath for 1 hour. Changes in the egg white portion in
each test tubes recorded.

OBJECTIVES:
1. To identify digestion as a process of hydrolysis; and
2. To determine the hydrolysis products of carbohydrate, fat and protein digestion.

LABORATORY OF CARBOHYDRATES
A. DIGESTION OF CARBOHYDRATES
Table 1. Hydrolysis of Starch
COLOR WITH IODINE
Time (min)
STARCH + SALIVA STARCH ONLY
2 Bluish Dark blue to black
4 Orange with blue Dark blue to black
6 Orange with blue Dark blue to black
 8 Orange with blue Dark blue to black
10 Orange with light blue Dark blue to black
12 Orange with yellow precipitate Dark blue to black
14 Orange with white precipitate Dark blue to black
Orange with light blue
16 Dark blue to black
precipitate
Orange with light blue
18 Dark blue to black
precipitate
Orange with light blue
20 Dark blue to black
precipitate

Table 2. Results of Benedict’s Test

COLOR WITH
SAMPLE
BENEDICT’S REAGENT IS GLUCOSE PRESENT?
Starch + saliva Greenish with green present (+) yes
Starch only Blue (-) no

1. Where does starch digestion begin?

Answer: The hydrolysis of starch begins in the mouth which is done by salivary amylase, but this is
only small in comparison to the extent performed by the pancreatic amylase in the small intestine.
2. What carbohydrate digestion occurs in the small intestine?
Answer: The breakdown of the longer carbohydrate chains facilitated by the pancreatic amylase and
the supporting role of maltase in the breakdown of the shorter chains already broken down from the
preceding processes.
3. What are the end products of carbohydrate digestion?
Answer: Glucose molecules are the end products of carbohydrate digestion.
4. Why do we need a digestive process?
Answer: Having a digestive process is essential because it is the only pathway available for obtaining
the important biomolecules that the body needs in order to survive. As has been discussed in this
laboratory report, the different processes observed make products that are essential for human life,
carbohydrates for energy, fats for proper nerve function, and proteins for functions such as in
antibodies

B. DIGESTION OF TRIACYLGLYCEROLS
Table 3. Bile Salts
MIXTURE OBSERVATIONS
Oil and water formation of two layers, oil on the top
Oil, water and bile salts Layers of oil becomes small shapes and circle.

Table 4. Hydrolysis by Lipase

Time (min) pH Volume of 0.1M NaOH (mL)
0 6.2 3.2 mL
20 6.4 1.5 mL
 40 6.2 2.2 mL
60 6.3 2 mL

1. What is the effect of bile salts on an oil and water mixture?

Answer: The bile emulsifies the oil into smaller globules and allows both of them to mix (formation of oil
droplets). This occurs in a mechanism similar to the concept of soap micelles.
2. What is the function of bile salts in the digestion of fats and oils (triacylglycerols)?
Answer: Bile salts act as a surfactant that aid in breaking down larger pieces of fat into smaller
manageable pieces which are more readily observed than when they are larger in size.
3. What products of lipase action would change the pH of a mixture containing triacylglycerol?
Answer: The production of free fatty acids that come about from lipase action on fats tend to change
the pH levels of the mixture by virtue of them being acidic in nature because of the carboxylic acid
group (-COOH)

C. PROTEIN DIGESTION
Table 5. Digestion of Protein
SAMPLE TEST TUBE
(Egg White) Water + HCl Pepsin + HCl Pepsin Only
Initial Appearance Solid big white Solid big white Solid big white
Final Appearance Solid big white Smaller big white Solid big white
Has any digestion
none yes none
taken place?
Hydrocholic acid
What caused the activated the pepsin to
N/A N/A
digestion process? be functional and this
where digestion start

5. Why does a person with low protein production of stomach HCl have difficulty with protein digestion?
Answer:This is because the lack or diminished amount of stomach HCl means that there are no
suitable conditions for the activation of pepsinogen into pepsin, which is the major protease in the
breakdown of proteins in the body. This leads to little to no protein digestion and could lead to various
bodily abnormalities.
6. What are the products of protein hydrolysis?
Answer: The end products of the protein hydrolysis are the amino acids that made them up.

DISCUSSION
` .
The first part of the experiment tackles about the digestion of carbohydrates specifically starch. The
starch solution was divided into two; one served as control the other was added with salivary amylase. Iodine
test was also used to indicate the presence of starch, which gives a blue-black color. Based on the results, the
solution with the starch and saliva gave a dark blue to black (of a positive iodine test) upon addition of iodine
every after two mins. But after 20 mins, the mixture no longer exhibited a blue-black color which indicates that
the starch was completely hydrolyzed. Below is a figure illustrating the hydrolysis of starch producing sugar
molecules with the aid of carbohydrase (amylase, maltase, etc).
Figure 1. Hydrolysis of starch producing sugar with the aid of of carbohydrase(amylase, maltase, etc).
Another test to confirm the complete hydrolysis of starch is the Benedict’s test. The positive result for this is the
presence of orange color in the solution, which indicates that glucose, the monosaccharide in starch, is
present. Based on the results, the solution containing saliva and starch gave a positive result which means that
glucose is present. The reaction equation is shown as:
=O
sugar – CH + 2 Cu2+ sugar – COH + Cu2O(s)

Figure 2. Reaction of cupric ion with a reducing sugar forming a red precipitate of cuprous oxide
The digestion of starch starts in the mouth, by means of the action of the salivary amylase, found in
saliva which actions for the breakdown of starch. Then it will be partially broken down into simpler portions in
the stomach. Hydrolysis then continues in the small intestine. The pancreas releases the enzyme pancreatic
amylase, which breaks the polysaccharide down into a disaccharide. The small intestine then manufactures
enzymes called lactase, sucrase and maltase, which break down the disaccharides into monosaccharides. The
monosaccharides are then absorbed in the small intestine. An illustration of the carbohydrate digestion is
shown in Appendix Figure 1.
The second part of the experiment is the digestion fats. This part was divided into two sections. The
first one was aided with Bile salts. Two solutions were formulated. The first was the oil and water mixture, and
the second was the oil, water, and bile salts mixture. The result was that, the oil and water in the first solution
formation of two layers, oil on the top. On the other solution, the mixtures mixed, and a soapy layer was
formed. The solution was also degraded. These differences in the two solutions give an explanation for the role
of the bile salts. Bile salts have detergent acts on particles of dietary fat which triggers fat globules to break
down or be emulsified into minute, microscopic droplets, enlarging the surface area of the fat, making it simpler
to digest.
The other was the digestion of fats in milk through the use of lipase (another enzyme vital to the
digestion of fat). Lipase hydrolyses the fat in milk to fatty acids which react with sodium hydroxide to lower the
pH of the mixture. This pH change is perceived by using phenolphthalein. Based on the results; as the length
of time increases, the value of pH also increases. The level of NaOH added was increasing in a very minute
amount ranging from 0.5 to 1.0 mL. The result implies that errors had occurred during the conduct of the
experiment. Various situations can imply for the unsuccessful result of the experiment such as the
temperature, impured reagents and not following the procedure carefully. Though errors are inevitable, certain
measurements must be observed to prevent such occurrence. But theoretically, the pH must be decreasing as
the amount of NaOH increases. Because, as time progresses, more and more of fats are converted into free
fatty acids. Also amino acids are amphoteric electrolytes and therefore improbable to produce the pH changes.
The production of both amino acids and fatty acids are presumed as they likely causes of the drop in pH.
Digestion of some fats can start in the mouth where lingual lipase breaks down some short chain lipids
into diglycerides, but mainly, lipid digestion proceeds in the small intestines. Lipids are emulsified by bile salts
which is made in the liver, deposited in the gall bladder and introduced into the small intestines. The pancreas
also adds the enzyme lipase which breaks down lipids. Intestinal lipase degrades the triacylglycerols. Other
fatty acids and other breakdown products are taken up by the intestinal mucosa and converted into triglycerols.
The triglycerols are then incorporated by the golgi apparatus into chylomicrons and sent through the lymphatic
system and bloodstream to tissues. The lipoprotein lipase then converts the triacylglycerol into fatty acid and
glycerol which is activated by the apoC-II in the capillary. The fatty acid then enters the cell and further
oxidized as fuel. The Fatty acid and Triglyceride Digestion is further illustrated in Appendix Figure 2.
 The last part of the experiment is the protein synthesis. Three set-ups were prepared: the first was
water and HCl, the second was pepsin and HCl, and the third was pepsin and water. The protein to be
assessed was albumin, the white portion of the hard-boiled egg. A piece of albumin was placed in each of the
test tubes, and the effect on albumin was observed. Based on the results, in the first set-up, the egg albumin
remained whole or compact, and the reason for this is the nonexistence of pepsin. In the second set-up, the
egg albumin was deformed or completely hydrolyzed. This is because of the presence of HCl which helps
degrade or hydrolyse the egg white. In the third set-up, the egg albumin slightly degraded, and this is because
of the pepsin. However, there was no HCl, unlike in the second set-up. As mentioned earlier, HCl helps in the
denaturation of the protein, so if HCl is not present, the protein will not be completely hydrolysed, because
pepsin will not be fully activated. The gastric juice in the stomach assists a lot in protein digestion, since it will
activate the pepsin, making the protein break down quicker into amino acids. So for a person with a minimal
production of gastric juice, they will certainly have trouble with protein digestion. Below is the general reaction
equation for the hydrolysis of proteins producing amino acids with the aid of proteases (pepsin, trypsin).

proteases (pepsin, trypsin)

proteases amino acids

Figure 3. General reaction equation for the hydrolysis of proteins with the aid of
Proteases (pepsin, trypsin).

Proteins are very large complex molecules that cannot be easily absorbed by the intestines. In order for
the proteins to be absorbed, they must be broken down to its simplest component, which is the amino acid.
Protein digestion begins in the stomach, in which the gastric juice and pepsin are found and are crucial as well.
The constituent of gastric juice is hydrochloric acid. It initiates denaturation of proteins, activates pepsinogen to
pepsin, and it makes pH in the stomach appropriate for the action of pepsin. The enzyme accountable for the
breakdown of proteins into amino acids is the pepsin.

IV. CONCLUSION
Therefore, digestion is an essential process in living organisms. It is in a form of hydrolysis process that
breaks down large complex biomolecules into simpler substances that can be easily absorbed by the
intestines. The hydrolysis products of carbohydrate, fat, and protein digestion are glucose molecules, glycerol
and fatty acids, and amino acids, respectively. Each product have its own metabolism which also has different
function such that carbohydrates are essential for energy storage; fats are for neural transmissions, and;
proteins are diverse that its functions counterpart almost all in the systems of the body.
V. ANSWERS TO QUESTIONS

7. Where does starch digestion begin?

Answer: The hydrolysis of starch begins in the mouth which is done by salivary amylase, but this is
only small in comparison to the extent performed by the pancreatic amylase in the small intestine.
8. What carbohydrate digestion occurs in the small intestine?
Answer: The breakdown of the longer carbohydrate chains facilitated by the pancreatic amylase and
the supporting role of maltase in the breakdown of the shorter chains already broken down from the
preceding processes.
9. What are the end products of carbohydrate digestion?
Answer: Glucose molecules are the end products of carbohydrate digestion.
10. Why do we need a digestive process?
Answer: Having a digestive process is essential because it is the only pathway available for obtaining
the important biomolecules that the body needs in order to survive. As has been discussed in this
laboratory report, the different processes observed make products that are essential for human life,
 carbohydrates for energy, fats for proper nerve function, and proteins for functions such as in
antibodies.
VI. REFERENCES
Garett, R.H and Grisham, C.M. 1963. Biochemistry. Addison-Wesley Publishing Company.
Nelson, David L. and Cox, Michael M. Lehninger, Principles of Biochemistry, Fourth
Edition: Study Guide and Solutions Manual. Marcy Osgood, University of New Mexico,
and Karen Ocorr, University of California, San Diego, 0-7167-5955-1. pp 238-261.
www.whfreeman.com/lehninger4e
www.bbc.co.uk/bitesize/ks3/science/organisms_behaviour_health/diet_drugs/revision/5/
www.functionalfitmag.com/blog/2011/12/21/digestion-101
www.lamission.edu/lifesciences/bio3labs/Bio3%20Lab05-Fa12-Enzymes.pdf
www.livestrong.com/article/417962-how-does-the-body-digest-carbohydrates/
www.news-medical.net/health/What-is-Digestion.aspx
www.open.edu/openlearn/science-maths-technology/science/biology/nutrition-proteins/content-section-1.7
www.vivo.colostate.edu/hbooks/pathphys/digestion/liver/bile.html