EXPERIMENT NO.

4
Enzymes and Digestion

Date Performed: September 19, 2019

Members:
TRIBO, Vincent Joshua, M.I.
TRINIDAD, Zarina Patricia, M.*
VILLAFLOR, John Emmanuel, M.*
YASUL, Dhaline Mae, F.
ZAUSA, Suzaine Shaine, M.I.
A. SALIVARY DIGESTION: FACTORS AFFECTING ENZYMATIC ACTIVITY
1. Effect of temperature

Plate 1. Results of Test tube A placed in an ice water bath, Test tube B placed at
37'C and Test tube C placed in a boiling water bath.
Specific Objective: To identify how temperature affects the rate of enzyme
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activity.
Result:

Test tube Temperature Color of the Solution Rank

A in ice water bath Dark violet 3

B 37'C Violet 2

C in boiling water bath Light violet 1

Principle: Most chemical reactions proceed at a faster rate as temperature

increases. The enzyme is a protein, and at high temperatures, the shape of
the protein is altered, preventing it from performing its function. It has been
denatured.

Performed by: Yasul, Dhaline Mae F.

2. Effect of pH

Plate 1. Result of test tube 4 by adding pH4 buffer solution, test tube 7 by
adding pH7 buffer solution and test tube 10 by adding pH10 buffer solution.
Specific objective: To identify the solution that indicates digestion of the
polysaccharide.
Result:
Table 1. Summary of Results of the

Test tube Color of solution Rank

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pH 4 Light red 3
pH 7 colorless 1
pH 10 Light blue 2
Principle: The rates of chemical digestion processes involving enzymes are
dependent on the pH of the surrounding fluids. The activity of amylase
changes as the pH changes. A higher enzyme activity will result in a faster
rate of reaction. Enzymes are globular proteins with a characteristic shape.
This shape can alter with changing pH, the result being a decrease in activity.

Performed by: Kayla Mae D. Diaz

3. Effect of Enzyme Concentration

Specific Objective: To identify

rate of time of digestion by using
different amount of enzyme concentation to the subtrate. Ranking the tubes
by 1- as the fastest digestion and 3- as the slowest digestion.
Result:

Test tube Content Color of the Solution Rank

(Theoretical Result)
1 1mL 1% starch TS, 1 drop of iodine Dark blue color (control)
solution.

2 1mL 1% starch TS, 1 drop of iodine Small amount of Colorless 3

solution and 3 drops of warm solution on top and
saliva reddish violet solution at
the bottom (10 seconds)
3 1mL 1% starch TS, 1 drop of iodine Moderate amount of 2
solution and 6 drops of warm Colorless solution on top
saliva and reddish violet solution
at the bottom(5 seconds)
4 1mL 1% starch TS, 1 drop of iodine large amount of Colorless 1
solution and 10 drops of warm solution on top and
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saliva. reddish violet solution at

the bottom(2 seconds)
Principle: The rate of an enzyme-catalysed reaction depends on the
concentrations of enzyme and substrate. As the concentration of enzyme is
increased, the rate of reaction or digestion increases. A dark blue solution
indicates the presence of starch. Partia digestion of starch result violet
or red colored solution. A colorless solution indicates the complete digestion of the
polysaccharide.
Performed by: Vincent Joshua V. Tribo
4. Effect of Substrate Concentration

Plate 1. Results of Test tube 1, 2, 3 and 4

Specific Objective: To identify how the substrate (starch TS) concentration
affect the rate of enzyme activity.
Result:

Test tube Content Color of the Solution Rank

(Theoretical Result)
1 1mL 1% starch TS, 1mL distilled Blue color 4
water and 1mL saliva

2 2mL 1% starch TS, 1mL distilled Green to yellow solution 3

water, and 1mL saliva

3 3mL 1% starch TS, 1mL distilled Orange solution 2

water, and 1mL saliva

4 5mL 1% starch TS, 1mL distilled Brick red precipitate 1

water, and 1mL saliva

Principle: The rate of an enzyme-catalysed reaction depends on the

concentrations of enzyme and substrate. As the concentration of substrate is
increased the rate of reaction increases.

Performed by: Yasul, Dhaline Mae F.

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5. Effect of Metal-ion Poisons on Enzymes Activity

Reference/s:
RSC. (accessed 2019, September 21). Enzymes. Retrieved from
www.rsc.org/Education/Teachers/Resources/cfb/enzymes.htm
Science Learning Hub. 2011, July 13. Rate of Digestion. Retrieved from
www.sciencelearn.org.nz/resources/1834-rate-of-digestion
Reference: https://www.sciencelearn.org.nz/resources/1834-rate-of-digestion?

B. PANCREATIC AND BILIARY DIGESTION

1. Pancreatic Amylase
Plate 1.
Specific Objective:
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2. Pancreatic Proteases

3. Pancreatic Lipase/Bile

EXPERIMENT NO. 5
DNA Extraction from Plant
Tissues

Date Performed: September 19, 2019

Members:
TRIBO, Vincent Joshua, M.I.
TRINIDAD, Zarina Patricia, M.*
VILLAFLOR, John Emmanuel, M.*
YASUL, Dhaline Mae, F.
ZAUSA, Suzaine Shaine, M.I.
 Page 7 of 18

EXPERIMENT NO. 6
Metabolism and Energy
Production
 Page 8 of 18

Date Performed: September 19, 2019

Members:
TRIBO, Vincent Joshua, M.I.
TRINIDAD, Zarina Patricia, M.*
VILLAFLOR, John Emmanuel, M.*
YASUL, Dhaline Mae, F.
ZAUSA, Suzaine Shaine, M.I.

1. Color, pH, and specific gravity

Plate 1.
Specific Objective:
Results:

Test Male urine Women urine

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Color Light amber Light yellow

pH pH 7 pH 7
Specific Gravity
Principle:
COLOR. Normal urine is straw yellow to amber in color. Abnormal colors include
bright yellow, brown, black (gray), red, and green. These pigments may result from
medications, dietary sources, or diseases. For example, red urine may be caused by
blood or hemoglobin, beets, medications, and some porphyrias. Black-gray urine
may result from melanin (melanoma) or homogentisic acid (alkaptonuria, a result of
a metabolic disorder). Bright yellow urine may be caused by bilirubin (a bile
pigment). Green urine may be caused by biliverdin or certain medications. Orange
urine may be caused by some medications or excessive urobilinogen (chemical
relatives of urobilinogen). Brown urine may be caused by excessive amounts of
prophobilin or urobilin (a chemical produced in the intestines).
In the presence of an indicator, colors range from deep blue-green in urine of low
ionic concentration through green and yellow-green in urines of increasing ionic
concentrations
pH.The double indicator principle gives a broad range of colors covering the entire
urinary pH range. Colors range from orange through yellow and green to blue.
Performed by: Credits to Group 4
2. Protein: Biuret test

Plate 1.
Specific Objective:to Identify the presence of peptide bond in
(albumin)protein in the urine samples.
Results::

Observation Male Urine Female Urine

Color of solution Blue Precipitate Blue precipitate
Interpretation Negative Negative
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(Positive or Negative)
Principle:
Biuret test is a general test for compounds having a peptide bond. Biuret is a
compound formed by heating urea to 180C. When biuret is treated with dilute
copper sulfate in alkaline condition, a purple colored compound is formed. This is
the basis of biuret test widely used for identification of proteins and amoniacids.
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 ie.

tripeptide gives positive biuret test.

The principle of biuret test is conveniently used to detect the presence of proteins in
biological fluids. Alkaline CuSO4 reacts with compounds containing two or more
peptide bonds to give a violet colored product which is due to formation of co-
ordination complex of cupric ions with unshared electron pairs of peptide nitrogen
and O2 of water.
Performed by: Credits to group 4
3. Glucose: Benedict’s Test

Plate 1.
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Specific Objective:: to Identify the presence of reducing sugars(glucose) in

the urine samples.
Results

Observation Male Urine Female urine

green solution with black
Color of solution Blue solution
and white precipitate
Interpretation Negative (traces of
Negative
(Positive or Negative) sugars)

Principle:Benedict's reagent starts out aqua-blue. As it is heated in the

presence of reducing sugars, it turns yellow to orange. The "hotter" the final color of
the reagent, the higher the concentration of reducing sugar. In general, blue to
blue-green or yellow-green is negative, yellowish to bright yellow is a moderate
positive, and bright orange is a very strong positive.

When Benedict’s solution and simple carbohydrates are heated, the solution
changes to orange red/ brick red. This reaction is caused by the reducing property
of simple carbohydrates. The copper (II) ions in the Benedict’s solution are reduced
to Copper (I) ions, which causes the color change.
The red copper(I) oxide formed is insoluble in water and is precipitated out of
solution. This accounts for the precipitate formed. As the concentration of reducing
sugar increases, the nearer the final colour is to brick-red and the greater the
precipitate formed. Sometimes a brick red solid, copper oxide, precipitates out of
the solution and collects at the bottom of the test tube.
Sodium carbonate provides the alkaline conditions which are required for the redox
reaction. Sodium citrate complexes with the copper (II) ions so that they do not
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deteriorate to copper(I) ions during storage.

Complex carbohydrates such as starches DO NOT react positive with the Benedict’s
test unless they are broken down through heating or digestion (try chewing
crackers and then doing the test). Table sugar (disaccharide) is a non-reducing
sugar and does also not react with the iodine or with the Benedict Reagent. Sugar
needs to be decomposed into its components glucose and fructose then the glucose
test would be positive but the starch test would still be negative
Performed by: Credits to group 4
4. Ketone Bodies: Legal’s Test

Plate 1.
Specific Objective: to test for ketone
bodies in the urine using legal’s test
Results:

Observation Normal Urine Specimen Abnormal Urine Specimen

Color of solution yellow yellow

Interpretation
negative negative
(Positive or Negative)
Principle:
The test is based on the principle of Legal`s test. Acetoacetic acid and acetone form
a violet coloured complex with sodium nitroprusside in alkaline medium.

Performed by: credits to Urine Analysis Lab Con | Medical Specialties |

Chemistry
 Page 13 of 18

5. Urobilinogen and Bile Pigments: Gmelin’s Test

Plate 1.
Specific
Objective::T o test
for Urobilinogen and bile pigments in the urine sample.
Results:

Observation Normal Urine Specimen Abnormal Urine Specimen

Formation of rainbow Formation of rainbow
color (Green, blue,red, color (Green, blue,red,
Color of ring
violet, etc.) at the violet, etc.)at the
junction. junction.
Interpretation
Positive result Positive result
(Positive or Negative)
Principle:
Concentrated Nitric acid as oxidizing agent is used to the experiment to
oxidize bilirubin and biliverdin giving different colors from green to violet.
Gmelin test is used to detect bile pigments in urine.
Performed By: Vincent Joshua Tribo
6. Blood: Guaiac Test

Plate 1.
Specific
Objective: to test the
presence of blood in the urine sample
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Results:

Observation Normal Urine Specimen Abnormal Urine Specimen

Color of solution Yellowish brown Yellowish brown
Interpretation
Negative Negative
(Positive or Negative)
Principle:
Guaiac test is used in determining blood either on stool or urine, In case of
urine guaiac test the heme portion of the hemoglobin reacts with guaiac solution
when nitrous acid acid was added that act as oxidizing agent. Which then produce
dark blue ring and when shaken a blue color solution was produce that indicates
the presence of blood in urine.
Performed by: Vincent Joshua V, Tribo
7. Electrolytes
a. Sodium Ion: Flame Test

Plate 1. Abnormal urine sample

(left) and Normal urine sample (right)
Specific Objective: to test the presence of iodine in the urine sample
Result

Observations Normal Urine Specimen Abnormal Urine Specimen

Formation of silver Formation of silver
Color of flame chloride, chloride,
Yellow to Light yellow Yellow to Light yellow
Interpretation
Negative Negative
(Positive or Negative)

Principle: The flame test is a procedure used in chemistry to detect the presence

of certain metal ions like sodium or potassium based on each element's characteristic
emission spectrum.The test involves introducing a sample of the element or compound
to a hot, non-luminous (blue) bunsen flame, and observing the color that results. The
formation of yellow color in the flames indicates the presence of sodium while purple
color flame indicate the presence of potassium. The intensity of color is directly
proportional to the amount of content of certain metal ion in the urine.
Performed by: Tribo, Vincent Joshua
 Page 15 of 18

b. Chloride Ion: Silver Nitrate Test

Plate 1. Abnormal urine sample (left) and Normal urine sample (right)
Specific Objective: to test for the presence of chloride in the sample
Result

Observations Normal Urine Specimen Abnormal Urine Specimen

Formation of silver Formation of silver
Color of solution chloride, chloride,
Yellow to Light yellow Yellow to Light yellow
Interpretation
Positive Positive
(Positive or Negative)
Principle:
The formation of white precipitate is due to the reaction of silver nitrate to
sodium chloride which is present in the urine. The amount of the white
precipitate form is directly proportional to the amount of chloride ion
present in the urine.
Performed by: Tribo, Vincent Joshua
8. Dipstick test

Plate 1.
Specific Objective:
Results:

Dipstick boxes Urine Urine sample Interpretation to Interpretation to

sample 1 2 result of sample 1 result of sample 2
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Yellow box(Protein) yellow yellow Negative in Negative in protein

protein presence presence
Orange box(pH level) Orange Orange pH 5 pH 5

Yellow box(Specific Gravity) Light yellow Dark yellow 1.020 1.030

Blue box(Glucose) blue blue Negative in Negative in glucose

glucose presence presence

 Principle: The test strips consist of a ribbon made

of plastic or paper of about 5 mm wide, plastic strips have
pads impregnated with specific chemicals that react with the
compounds present in urine producing a characteristic
color(Chromatography). For the paper strips the reactants are
absorbed directly onto the paper. Paper strips are often specific
to a single reaction (e.g. pH measurement), while the strips with
pads allow several determinations simultaneously.Hence this
test method is based on Lateral flow chromatography. It’s dipped
into your urine, and the chemicals on the stick react and change
color if levels are above normal. Things the dipstick test can check
for include:
 Acidity, or pH. If the acid is above normal, you could have kidney
stones, a urinary tract infection or another condition.
 Protein. This can be a sign your kidneys are not working right.
Kidneys filter waste products out of your blood, and your body needs
protein.
 Glucose. A high sugar content is a marker for diabetes.
 White blood cells. These are a sign of infection.
 Bilirubin. If this waste product, which is normally eliminated by
your liver, show up, it may mean your liver isn’t working properly.
 Blood in your urine. Sometimes this is a sign of infections or
certain illnesses.

Performed by: Tribo, Vincent Joshua

https://mltgeeks.com/urine-dipstick-principle-and-significance-of-
 Page 17 of 18

parameters/

https://www.nku.edu/~whitsonma/Bio150LSite/Lab
%203%20Organic/Bio150LRevMolec.html#targetText=In%20general%2C%20blue%20to
%20blue,(See%20below).&targetText=Water%20plus%20Benedict's%20reagent
%20is,result%20(no%20sugar%20present).
https://microbiologyinfo.com/benedicts-test-principle-composition-preparation-
procedure-and-result-interpretation/
https://www.onlinebiologynotes.com/biuret-test-principle-requirements-
procedure-and-result/
http://www.lifebridgehealth.org/uploads/public/documents/lab/nw-clinitek-
status-plus-urine-dipstick-testing.pdf

Reference/s:
Cite your references in APA format
 Page 18 of 18

Last name, F. M. (Year, Month Date Published). Article title. Retrieved from URL
Example:
Arellano, A. C. (2018, May). Platelet-Increasing Activity of the Formulated Syrup
from the Expressed Juice of the Leaves of Papaya (Carica papaya). Retrieved from
www.ijppr.com
If you got the answer from a book, no need to cite the URL.
REMINDERS:
1. DO NOT CHANGE THE FONT, FORMAT AND PAGINATION.
2. All results should be tabulated.
3. Plates and tables should be labeled accordingly. The next photo should be
labeled Plate 2, next table should be labeled Table 2, so on and so forth.
4. All lab reports should be saved in PDF format before sending them via email.
5. The group leader should send the report. All groupmates should be in Cc section.
6. If a groupmate is absent on a certain experiment performed, mark their names
with an asterisk at the front page (e.g. ARELLANO, Alexis C.*)
7. Deadline for submission is September 16, Monday, 11:59 PM. Late reports will no
longer be accepted.
8. All the references used should be listed alphabetically based on the last name of
the author. These will be on the last page after all the results