The Galaxy School

IB Biology Lab
Experiment : To investigate the effect of lipase on milk with and without bile.
Material required:
Milk treated with bile, Untreated or raw milk, lipase solution, bile salt solution,
Data logger, pH sensor, sodium bicarbonate.

Procedure :
(a) Label three beakers 1-3.

(b) Using a graduated syringe, place 10 cm3 of untreated or raw milk in beaker no.
2 and 10 cm3 of bile treated milk in beaker 1 and 3 each.(10ml of milk + 5 ml bile)
(c) Rinse the pipette or syringe and use it to place 6 cm3 diluted sodium carbonate
solution (which is provided to you) in each beaker. This solution is to make the
mixture alkaline.
(d)Connect a pH probe to a data logger and switch on the data logger. Calibrate the
probe with known buffer if needed.
(e) Place the pH probe in beaker 1, set up a program that the probe takes reading at
every 2 mins till 30 minutes and touch the start key of the data logger. Shake the
beakers in between to mix the contents.
(f) Note the reading in the table below upto 30 mins.
(g) Rinse the pH probe with distilled water.
(h) With the graduated pipette or syringe, place 5 cm3 of 2% lipase solution (or 5%
lipase) in beaker 2.
(i) Place the pH probe in beaker 2 and start taking reading. Note down the result in
table below.
(j) Repeat the steps from (g) to (i) for beaker 3.

Result:
Time (min)

Beaker 1
Milk + Bile

Beaker 2
Beaker 3
Milk + Lipase Milk + Bile +Lipase
pH of the solution

0
4
8
12
16
20
24
28
30
Difference in pH
(Final Initial pH)

Review questions
1. Why does fat digestion affect the pH of a solution?
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2. In which beaker did fat digestion occur most rapidly?
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3. Do bile salts digest fats? What do bile salts do?
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4. Explain what emulsification means. How does emulsification
aid in fat digestion?
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Experimental Set Up:

Note down initial pH of milk

As time goes on pH of milk mixed with lipase and bile is decreasing

pH of the milk at 30th min.

NOTE : Yeast is very good source of lipase enzyme.