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Document: LABORATORY MANUAL

Title: CHM 311 Biochemistry Laboratory

EXPERIMENT 6

ENZYMATIC CATALYSIS OF THE HYDROLYSIS OF FATS (ACTION OF

PANCREATION ON BUTERFAT)

OBJECTIVES: At the end of the experiment, students are expected to learn how to:

1. Explain the digestive function of pancreatin in the hydrolysis of fats.

2. Describe the condition under which fat is hydrolyzed with pancreatin.

ASSIGNMENT

1. Explain the function of sodium carbonate solution on the digestion of butterfat by pancreatin.

It acts as a catalyst for releasing enzymes from the pancreatin so it can break down carbs, lipids, or protein
(mainly starch, though). Catalysis is the process of increasing the rate of a chemical reaction by adding a substance
known as a catalyst. Catalysts are not consumed in the catalyzed reaction but can act repeatedly. Often only very
small amounts of catalyst are required.

2. Name three enzymes in pancreatin and state the function of each.

Exocrine glands in the pancreas create enzymes that aid in digestion. These enzymes include trypsin and
chymotrypsin for protein digestion, amylase for carbohydrate digestion, and lipase for fat digestion.

3. Write the structure of a typical molecule of butterfat and write the equation for its digestion
with pancreatin.

DISCUSSION
The pancrease is a large gland that lies in the upper abdomen, behind the lower part of the
stomach. It serves two major functions. One is to produce the hormones insulin and glucagon,
which help regulate metabolism. The other is to produce pancreatic juice, a secretion delivered
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Revision Date: NOVEMBER
2021
Document: LABORATORY MANUAL

Title: CHM 311 Biochemistry Laboratory

into the upper part of the small intestine (duodenum), where it aids digestion.

Pancreatin is a mixture of the fat dissolving enzyme, lipase, the protein enzymes such as
protease, and those that break down carbohydrates like amylase. The enzymes in pancreatin may
come from pork or beef. If you are vegetarian or have allergies or religious restrictions to any of
these items, you should know this, Pancreatic enzymes are not standardizes, and contain a
variety of enzymes, including trypsin, chrmotrypsin, lipase, amylase, colipase, and ribonuclease.

These enzymes catalyze the hydrolysis of fats into glycerol and fatty acids, protein into proteoses
and derived substances, and starch into dextrins and sugars.
REAGENTS AND APPARATUS

Reagents/Materials:

Milk Cream 1% sodium carbonate

solution
Litmus Litmus paper (red and 2% pancreatin solution
solution blue)
Distilled
water

Apparatus: Test tubes, test tube rack, test tube brush, test tube holder, Bunsen burner, tripod,
wire gauze, water bath, thermometer, beakers, stirring rod, graduated cylinder (10-mL).

PROCEDURE

Pancreatin is made by drying the pancreases of cattle and other animals. When placed in
solution or suspension it contains a mixture of enzymes. The solution is not stable for long
periods of time, and should be freshly prepared and stored in a refrigerator when not in use.

1. Fill a test tube about 1/3 full of “half and half” a mixture of milk and cream.
2. Add 1 ml of 1% sodium carbonate solution and 3 ml of litmus solution to the test
tube.
3. Mix thoroughly and divide the mixture equally between two test tubes which may be
labeled A and B.
4. To tube A, add 2 ml of 2% pancreatin solution, and to tube B add 2 ml of distilled
water.
5. Mix the contents of the tubes and place them in a water bath maintained at a
temperature of approximately 400C. Note any change in color of the tubes.
6. Let them remain in the water bath for 30 minutes or until a definite color change is
noted in one of the tubes.
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Title: CHM 311 Biochemistry Laboratory

7. Also note any change in odor of one or both of the tubes. Explain your observations.
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Document: LABORATORY MANUAL

Title: CHM 311 Biochemistry Laboratory

DATA/RESULTS

Observatio With Pancreatin Without Pancreatin

ns
Color yellow

Odor Foul Odor waxy

CONCLUSION (Give explanations for all your observations)

The reaction may be tracked by noticing how the pH changes over time. Pancreatin, a crude
extract of bovine pancreas including amylase, trypsin, lipase, ribonuclease, and protease, was employed
as the source of lipase in this investigation. It's a digestive aid that's also utilized in the manufacture of
predigested foods. The experiment demonstrated how fat dissolves.