Pancreatic

Lipase and Bile

Group 6

Pancreatic Lipase and Bile

Objective:
To describe the role of enzymes in the digestive
process
Reagents used:
Vegetable oil
Pancreatin
Sodium choleate
Distilled water

Pancreatic Lipase and Bile

Pancreatic Juices:
contains enzymes such as amylase, lipase and
trypsin
aid in digestion
Lipase
an enzyme released by the pancreas into the
small intestine that helps the body to absorb and
breaking down fats
Bile
secreted by the liver that is important in
absorbing fat-soluble substances
Pancreatin

Pancreatic Lipase and Bile

Methodology:
Contents

Test tube
1

Test tube
2

Test tube
3

Vegetable
Oil

20 drops

20 drops

20 drops

Pancreatin

3.0 mL

Sodium
Choleate
Distilled
water

3.0 mL
3.0 mL

3.0 mL

3.0 mL

3.0 mL

Pancreatic Lipase and Bile

Methodology:
B. Mix the contents of all tubes thoroughly
C. Note to initial pH using pH paper. Record
the initial pH. Adjust the initial pH to 7 (or a
little bit higher) by adding 0.1% NaOH
Dropwise
D. Immerse all the tubes in a water bath set
at 37C for 1 hour. Check the pH of your
solutions after every 15 minutes. Remove all
tubes from the water bath at the same time.
Record the final pH

Pancreatic Lipase and Bile

Before water bath:

Without
NaOH; pH:
5

With 0.1%
NaOH; pH:
8

Pancreatic Lipase and Bile

During water bath:
1st 15
minutes

Next 15
minutes

Pancreatic Lipase and Bile

During water bath:
Next 15
minutes

Final15
minutes;
pH:6

Pancreatic Lipase and Bile

Setup and Final
solution:

Pancreatic Lipase and Bile

Results:
Content
Pancreatin +
Vegetable oil

Observations
Solution change
color indicating
partial digestion

Vegetable oil + Bile

solution
Vegetable oil + Bile
Solution
solution +
drastically change
Pancreatin
color indicating
complete
digestion

Pancreatic Lipase and Bile

Sodium Choleate
bile acid formed by bacterial action from cholate
and conjugated with glycine or taurine
Functions of each reagents:
Pancreatin Solution
- Contains lipases and is used to stimulate
pancreatic lipase.
Human Pancreatic Lipase
- Converts triglyceride substances found in
ingested oils to monoglycerides and free fatty
acids

Pancreatic Lipase and Bile

Principle behind the

determination of the
solutions pH before and after
incubation
The pH of a solution would
decrease as free fatty acids are
produced from fats; therefore,
reduction in pH would imply that

Pancreatic Lipase and Bile

Chemical Equations involved:

Pancreatic Lipase and Bile

Application of Sodium Choleate:

- Trihydroxy bile salt that is used as a
digestive aid in dietary supplements and
conjugated to pancreatin
- Deoxycholic acid acts a detergent to
solubilize fats for intestinal absorption, is
reabsorbed itself, and is used a as
choleretic and detergent

Pancreatic Lipase and Bile

Purpose of adjucting the pH
- Changes in pH affects the shape of
enzyme
- It also the change the shape or charge
properties of the substrate so that either
the substrate cannot bind to the active
site or it cannot undergo catalysis