LAB REPORT #4: DIGESTIVE SYSTEM

Biology I, Pre-Health Sciences – Durham College, Fall 2020

/40 marks

EVALUATION NOTES

Read before submitting:

 The lab report is worth 5% of your final grade.

 This lab report is due by 11:59 pm on the due date established by your instructor.

a. A report submitted after this due date is considered late. Late submissions
are penalized 10% per day up to a max of 3 days (including weekends).
After that a grade of 0 is assigned.

 This lab report should be submitted online to the DC Connect Assignments folder.

a. The report should be submitted to DC Connect. Unless otherwise directed,

submissions made by other means (E.g. email or hard copy) will not be
accepted and will result in a grade of 0.

b. Computer malfunctions or technical issues are not accepted as excuses for

late or missing reports. TIP: Submit your report early so that you have time
to resolve any potential technical issues prior to the deadline.

 While students may work in groups to complete labs, each student must submit
their own original work.

1
PART 1 – USING A MICROPIPETTE

Introduction
When working in the lab, it is an
important skill to learn how to use a
micropipette. This is an instrument
capable of transferring extremely small
volumes (microliters = L) of liquid. It is
often used when working with enzymes
or other reagents that are very
expensive. While we are not able to use
actual micropipettes in an online
environment, it is still your responsibility
to learn how one works in theory.

[]

Procedure
1. To learn how to use a micropipette, inspect the video, images, chart and sample
procedure below:

a. Overview video about micropipettes:

https://www.youtube.com/watch?v=Jfqafjt4q6U

b. Image of various micropipettes and corresponding tips [Source].

2
 c. Chart showing 3 sizes of micropipettes – P20, P200 and P1000. Note
how the dials can be adjusted to measure different volumes of liquids.
Micropipett
P20 P200 P1000
e
Lowest
2 µL 20 µL 200 µL
Volume

Highest
20 µL 200 µL 1000 µL
Volume

Tens 0 Hundred Thousand

s 0 s 0 This
Dial Ones 5 This is This is is
5.5 µL Tens 5 55 µL Hundreds 5 550
Tenth
s 5 Ones 5 Tens 5 µL

d. Sample procedure for using a micropipette to measure 10.0 L:

o Set the volume of a P20 to 10.0 L 1

 Adjust the dial at the top of the micropipette to 0
0
o Load a clean tip
 Attach a tip by placing the end of the micropipette firmly onto the
top of any clean tip from the tip box
 The tip should remain “stuck” after you remove it from the box

o Load the sample

 Push the plunger down slowly to the point of first resistance
 Place the tip so that it is emerged in the sample that you wish to
transfer
 Release the plunger slowly to draw up the liquid
 Visually inspect that no bubbles are present

o Deliver the sample

 Place the loaded tip into the receiving vessel
 Depress the plunger to the point of second resistance to expel
all the liquid. All of the liquid should be removed at this point.
 Keeping the plunger depressed, withdraw the tip

o Eject the tip

 Press the discharge button on the back of the micropipette
 Place the used tip into the waste receptacle

3
 2. (5 marks) Use the information in Question 1 to answer the multiple choice
questions below. Highlight the answer that you think is correct.

a. Which of the following micropipettes should be used when measuring 850

L of liquid in the lab?
i. P20
ii. P200
iii. P1000

b. Which of the following micropipettes should be used when measuring 8.5

L of liquid in the lab?
i. P20
ii. P200
iii. P1000

c. If a P20 has a dial setting as shown, how many L would it be measuring? 0

i. 850 L 8
ii. 85 L 5
iii. 8.5 L
iv. 0.85 L

d. If a P1000 has a dial setting as shown, how many L would it be measuring? 0

i. 2000 L 2
ii. 200 L 0
iii. 20 L
iv. 2 L

e. There are 2 resistances that you can feel when pressing down the plunger
of a micropipette. When should you press the plunger down to the second
level of resistance?
i. When you are attaching the pipette tip
ii. When you are loading liquid into the pipette tip
iii. When you are delivering liquid from the pipette tip
iv. When you are ejecting the pipette tip
v.

4
PART 2 – GUESS THAT ENZYME

Introduction
Imagine that you are working in a nutrition
lab at Health Canada. It is your job to
perform various quality control checks on
common food items to make sure that the
ingredients on the food label match the
ingredients in the actual food.
Unfortunately, you accidently mislabelled 3
digestive reagents commonly used in your
job - amylase, peptidase, and a digestive
cocktail containing bile, lipase, peptidase
and amylase. As these reagents are very
expensive, you feel anxious to fix your Uh oh…what is what? [source]
mistake and identify which vial is which.
Luckily, you remember that you are familiar with Benedict’s test, Biuret’s test and
Cabbage Indicator test [Refer to Lab 1 to review Benedict’s test and Biuret’s test]. To
your delight, you realize that these tests can help you to identify which vial is which.
Complete the procedure below to fix your mistake…and hopefully avoid being fired!

5
Procedure
1. To solve the riddle above, let’s first REVIEW a few important concepts.

a. (1.5 marks) Examine the figure below about Benedict’s test and then use it
to fill in the blanks below. [Refer back to Lab 1 if you get stuck.]

Fill in the blanks: Benedict’s test is used

to detect if a solution contains
carbohydrates. Specifically, it can detect 2
types of carbohydrates - the ________
monosaccharides _____________ and
the ________ disaccharides
_____________ .If the solution contains
these carbohydrates, the solution will be
green/yellow/orange/red at the end of the
test (a positive result). If it does NOT
contain these carbohydrates, the solution
will be ________Blue___________ at the
end of the test (a negative result).

b. (1.5 marks) Examine the figure below about Biuret’s test and then use it to
fill in the blanks below. [Refer back to Lab 1 if you get stuck.]

Fill in the blanks: Biuret’s test is used to

detect if a solution contains proteins. If a
solution contains _____amino
acids______________, it will be the colour
pink at the end of the test (a positive
result). If the solution contains
__polypeptides_________________, it will
be the colour purple at the end of the test
(also a positive result). If the solution does
not contain proteins at all, it will be the
colour ___blue_________ (a negative
result).

c. (0.5 marks) Examine the figure below about cabbage indicator test and
then use it to fill in the blanks below. [This is a new type of test!]

Fill in the blanks: Cabbage indicator test

is used to detect if a solution contains fatty
acids. If a solution contains fatty acids it
will be the colour ____blue_____ at the

6
 end of the test (a positive result). If the
solution does not contain fatty acids, it will
be the colour red/purple (a negative
result).

d. (4.5 marks) Recall that the mixed up bottles of digestive agents contain
lipase, amylase, peptidase, and bile. Describe their roles in digestion by
completing the chart below. Be as specific as possible. The first line of the
chart is completed for you. [Refer back to your online module for Week 9 if
you get stuck.]

Enzyme or
How does this break down food?
chemical?
This breaks down EMULSIFIED
Lipase Enzyme TRIGLYCERIDES
into MONOGLYCERIDES + FREE FATTY ACIDS

This breaks down

Amylase ___Enzyme_______ _polysaccharides______________
into ___disaccharides___________
This breaks down _______smaller
Peptidas polypeptides_________________
__Enzyme________
e into ________amino
acids___________________________
This breaks down ___lipid
droplets_____________________
__Chemical_______
Bile into _____emulsified
_
triglycerides____________________________
__

2. Next, let’s continue solving the riddle by completing a set of EXPERIMENTS.

*NOTE: Typically you would be in the lab completing this section yourself with
micropipettes. For this online course, this will be done for you using the
following procedure:

a. Recall from the “Introduction” that 1 bottle contains amylase, 1 bottle contains
peptidase, and 1 bottle contains a combination of bile, lipase, peptidase and
amylase…but we don’t know which is which. To keep them organized for
this experiment the agents were temporarily called “X,” “Y,” and “Z.”

b. Next, we tested whether X, Y and Z were capable of digesting various

macromolecules. To do this, starch (a polysaccharide), albumin (a complex

7
 protein), and vegetable oil (a triglyceride) were mixed with X, Y or Z and
allowed to incubate for 1 hour. Then, Benedict’s test, Biuret’s test, and
Cabbage Indicator test was carried out. See the grid below to visualize the
12 different experiments (A1-A4, B1-B4, C1-C4):
A1 A2 A3 A4
Benedict’ Starch Starch Starch Starch
s test +X +Y +Z
B1 B2 B3 B4
Biuret’s Albumin Albumin Albumin Albumin
test +X +Y +Z
Cabbage C1 C2 C3 C4
indicator Oil Oil Oil Oil
test +X +Y +Z

c. Next, the results of the experiment were examined as shown below:

Benedict’
s test

Biuret’s
test

Cabbage
indicator
test

3. (3 marks) Given the information in Questions 1 and 2, fill in the chart below by
providing the following:

For Benedict’s test (A1-A4): Write “YES” if mono or disaccharides are present
For Biuret’s test (B1-B4): Write “YES” if amino acids are present
For Cabbage test (C1-C4): Write “YES” if fatty acids are present

A1 A2 A3 A4
Benedict’
s test NO NO YES YES
Biuret’s B1 B2 B3 B4
test

8
 YES YES YES YES

C1 C2 C3 C4
Cabbage
indicator NO NO YES NO
test
4. (3 marks) It’s now time to solve the riddle and assign proper labels for X, Y and
Z. Use the data from Question 3 to fill in the chart below. Justify your choice
using at least one piece of evidence from the experiment.
.
List X, Y Unlabeled Evidence to support your choice
or Z Bottle

- Monosaccharides and disaccharides were present

as well as
Z amylase
polypeptides

- Only amino acids were present

X peptidase

cocktail of
- All the experiments were
bile, lipase,
a positive result which
Y peptidase
means all were present
and
amylase

9
PART 3 – DIGESTING MOLECULES OF LIFE: FROM FOOD TO FUEL

Introduction
In this final lab section, you will create a study tool that will help you to learn the various
digestive enzymes and chemicals that digest the 4 molecules of life – carbohydrates,
proteins, nucleic acids, and lipids.

Procedure
1. Imagine that you just ordered a delicious take-out meal: sushi. For simplicity,
imagine that this meal only contains triglycerides, polysaccharides, complex
proteins, and nucleic acids. As you eat, also imagine that these macromolecules are
broken down into smaller pieces as digestive enzymes and chemicals are released
from various parts of the digestive system. The figure below shows these
macromolecules in their original, partially digested, and fully digested forms. Notice
that each one is associated with a letter from A-L.

LIPIDS CARBOHYDRATES PROTEIN NUCLEIC ACID

A D G J

B E H K

C F I L

10
2. Using your knowledge of digestion from your online learning this week, track the
sushi through the digestive system. To do this, fill-in the charts below for
polysaccharides, triglycerides, nucleic acids and complex proteins.
NOTES:
- If no digestion takes place in a particular region, just leave it blank in the chart
- The tracking of a polysaccharide has been completed for you. Use it as an
exemplar for completing the rest of the charts.

a. Tracking a polysaccharide (EXEMPLAR):

Describe the food Describe the food List the name of List the name of
molecule as it molecule as it the enzyme or the organ that
ENTERS this part EXITS this part chemical makes the
involved enzyme or
chemical
MOUTH What does it look What does it look Salivary Salivary gland
like (A-L)? like (A-L)? amylase

D E
What is its name? What is its name?

Polysaccharide Disaccharide
STOMACH What does it look What does it look
like (A-L)? like (A-L)?

What is its name? What is its name?

SMALL What does it look What does it look

INTESTINE like (A-L)? like (A-L)?
(liver/gall
bladder)
What is its name? What is its name?

SMALL What does it look What does it look Pancreatic Pancreas

INTESTINE like (A-L)? like (A-L)? amylase
(pancreas)
D E
What is its name? What is its name?

Polysaccharide Disaccharide
SMALL What does it look What does it look Maltase Brush border
INTESTINE like (A-L)? like (A-L)? Sucrase (small intestine)
(brush Lactase
border) E F
What is its name? What is its name?

Disaccharide Monosaccharide

True or False: Digested carbohydrates get absorbed into the body via a blood capillary

11
 b. (6+1 marks) Tracking a triglyceride:

Describe the food Describe the food List the name of List the name of
molecule as it molecule as it the enzyme or the organ that
ENTERS this part EXITS this part chemical makes the
involved enzyme or
chemical
MOUTH What does it look What does it look
like (A-L)? like (A-L)?

What is its name? What is its name?

STOMACH What does it look What does it look

like (A-L)? like (A-L)?

What is its name? What is its name?

SMALL What does it look What does it look Bile Liver

INTESTINE like (A-L)? like (A-L)?
(liver/gall
bladder) A B
What is its name? What is its name?

Lipid droplets Emulsified

triglycerides
SMALL What does it look What does it look Lipase Pancreas
INTESTINE like (A-L)? like (A-L)?
(pancreas)
B C
What is its name? What is its name?

Emulsified Monoglycerides + 3
triglycerides fatty acids
SMALL What does it look What does it look
INTESTINE like (A-L)? like (A-L)?
(brush
border)
What is its name? What is its name?

True or False: Digested lipids get absorbed into the body via a blood capillary

12
 c. (6+1 marks) Tracking a nucleic acid:

Describe the food Describe the food List the name of List the name of
molecule as it molecule as it the enzyme or the organ that
ENTERS this part EXITS this part chemical makes the
involved enzyme or
chemical
MOUTH What does it look What does it look
like (A-L)? like (A-L)?

What is its name? What is its name?

STOMACH What does it look What does it look

like (A-L)? like (A-L)?

What is its name? What is its name?

SMALL What does it look What does it look

INTESTINE like (A-L)? like (A-L)?
(liver/gall
bladder)
What is its name? What is its name?

SMALL What does it look What does it look Nuclease Pancreas

INTESTINE like (A-L)? like (A-L)?
(pancreas)
J K
What is its name? What is its name?

Nucleic Acid Nucleotides

SMALL What does it look What does it look Nucleosidase Small Intestine
INTESTINE like (A-L)? like (A-L)?
(brush
border) K L
What is its name? What is its name?

Nucleotides Sugar, Phosphate,

base

True or False: Digested nucleic acids get absorbed into the body via a blood capillary

13
 d. (6+1 marks) Tracking a complex protein:

Describe the food Describe the food List the name of List the name of
molecule as it molecule as it the enzyme or the organ that
ENTERS this part EXITS this part chemical makes the
involved enzyme or
chemical
MOUTH What does it look What does it look
like (A-L)? like (A-L)?

What is its name? What is its name?

STOMACH What does it look What does it look Pepsin Stomach

like (A-L)? like (A-L)?

G H
What is its name? What is its name?

Complex proteins Smaller

polypeptides
SMALL What does it look What does it look
INTESTINE like (A-L)? like (A-L)?
(liver/gall
bladder)
What is its name? What is its name?

SMALL What does it look What does it look Trypsin Pancreas

INTESTINE like (A-L)? like (A-L)?
(pancreas)
G H
What is its name? What is its name?

Complex proteins Smaller

polypeptides
SMALL What does it look What does it look Peptidase Small intestine
INTESTINE like (A-L)? like (A-L)?
(brush
border) H I
What is its name? What is its name?

Smaller Amino acids

polypeptides

True or False: Digested proteins get absorbed into the body via a blood capillary

14