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Experiment 7 - Isolation of Casein and Lactose From Milk

This document describes an experiment to isolate the proteins casein and lactose from milk. The procedure involves: 1) Dissolving powdered skim milk in warm water and adding acetic acid to precipitate the casein protein. 2) Adding ethanol to the remaining liquid to precipitate the lactalbumin and lactoglobulin proteins. 3) Heating the liquid to near boiling to further precipitate these proteins, then allowing lactose crystals to form upon cooling and standing. The purpose is to separate the main components of milk - the proteins casein, lactalbumin and lactoglobulin, and the sugar lactose.

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Mikaela Jasmine Morales
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6K views4 pages

Experiment 7 - Isolation of Casein and Lactose From Milk

This document describes an experiment to isolate the proteins casein and lactose from milk. The procedure involves: 1) Dissolving powdered skim milk in warm water and adding acetic acid to precipitate the casein protein. 2) Adding ethanol to the remaining liquid to precipitate the lactalbumin and lactoglobulin proteins. 3) Heating the liquid to near boiling to further precipitate these proteins, then allowing lactose crystals to form upon cooling and standing. The purpose is to separate the main components of milk - the proteins casein, lactalbumin and lactoglobulin, and the sugar lactose.

Uploaded by

Mikaela Jasmine Morales
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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Experiment 7

ISOLATION OF CASEIN AND LACTOSE FROM MILK

Introduction:

Milk is an opaque white or bluish-white liquid secreted by the mammary glands of female
mammals, serving for the nourishment of their young. It is the most nutritionally complete food
found in nature. The amounts of these nutrients present in different types of milk differ greatly,
however. Cows' milk and goats' milk are almost identical in every respect. Human milk contains less
than half of the proteins and minerals of cows' or goats' milk, but almost 1.5 times as much sugar.

There are three kinds of proteins in milk: caseins, lactalbumins, and lactoglobulins. All three
are globular proteins, which tend to fold back on themselves into compact, nearly spheroidal units
and are more easily solubilized in water as colloidal suspensions than fibrous proteins are. They are
"complete proteins", so- called because they contain all the amino acids essential for building blood
and tissue, and they can sustain life and provide normal growth even if they are the only proteins in
the diet. These proteins not only contain more amino acids than plant proteins, but they contain
greater amounts of amino acids than the proteins in eggs and meats. Casein, the main protein in
milk, is a phosphoprotein, meaning that phosphate groups are attached to the hydroxyl groups of
some of the amino acid side-chains. Casein exists in milk as the calcium salt, calcium caseinate. It is
actually a mixture of at least three similar proteins, which differ primarily in molecular weight and
the amount of phosphorus groups they contain.

Materials:

Triple beam balance, watch glass, 2 graduated cylinder (10mL and 50mL), beaker
(100mL), beaker (250mL), thermometer, electric stove, dropper, glass rod, wire gauze, spatula

Things to Bring:
10 grams non-fat or skimmed milk

Reagents:

10% acetic acid, 95% ethanol, 1:1 diethyl ether-ethanol mixture

Procedure:

A. Isolation of casein

Weigh out 5 grams of powdered non-fat or skimmed dry milk and dissolve it in 20 mL of
warm water in a 100 mL beaker. Bring the temperature of the solution to 55 oC (do not exceed) on a
hot plate, remove the thermometer, and then add dropwise a solution of 10% acetic acid while
stirring with a stirring rod. Do not add the acid too rapidly. Continue the acid addition (slightly less
than 2 mL will be required), keeping the beaker on the hot plate, until the liquid changes from milky
to almost clear and the casein no longer separates. It is important not to add too much acid, because
it may hydrolyze some of the lactose in the milk and reduce your yield. Stir the precipitated casein
until it forms a large amorphous mass; then remove it with a stirring rod or tongs and place it in
another beaker.

Immediately add 35mL of 95% ethanol to the original beaker containing the remaining liquid, stir for
a few minutes, and save the resulting mixture for the later separation of lactose below. The
separation of lactose should be done as soon as possible during the same laboratory period.

Collect the casein by filtration to remove as much water as possible. Press the solid with a spatula.
Place the casein in a 100 mL beaker and add 5 mL of acetone (CAUTION: HIGHLY FLAMMABLE - NO
FLAMES). Stir the casein in the ether-ethanol mixture for 5 minutes, decant ether-ethanol, and
repeat the process with a second 5 mL of ether-ethanol. After the second washing with ether-
ethanol, filter the product. The ether- ethanol washings remove any small quantities of fat that may
have precipitated with the casein. Place the casein between several layers of paper towels to help
dry the product, and let it stand in the air for 10-15 minutes. Divide the wet product in half, and
weigh the two portions. Place one portion in a 125 mL Erlenmeyer flask with 35 mL of water and 0.5
mL of 1M NaOH, stopper the mixture, shake it to ensure solution of as much of the casein as
possible, and save it for use in the chemical tests on the future experiment on protein.

Calculate the total yield of casein from the powdered milk. Show your calculations.

Isolation of Lactose:

Gently heat to near boiling the liquid with Ethanol. Bumping will not be a problem so long as you stir
the solution constantly and vigorously with a glass rod. The solution will foam somewhat as it
refluxes. This procedure precipitates the remaining proteins lactalbumin and lactoglobulin. While the
solution is still warm, transfer to centrifuge tubes and centrifuge for 5 minutes. Decant the liquid
back into the 50-ml Erlenmeyer flask, stopper and let stand until the next lab period. Lactose crystals
should slowly form in the solution as it stands.

Data:

Describe the isolated casein and lactose:

Conclusion:

Youtube Link: https://www.youtube.com/watch?v=qzfFajukhTU

Questions to answer:

1. What component of milk is responsible for its white colour?

2. Why is lactose much more soluble in water than in ethanol?

3. What is the purpose of 10% acetic acid to the milk colloidal suspension?

4. What is the purpose of adding 1:1 ether-ethanol mixture to casein precipitates?


Experiment 7
ISOLATION OF CASEIN AND LACTOSE FROM MILK

Group #: _________
Group Members:
_______________________________________
_______________________________________
_______________________________________
_______________________________________
_______________________________________

Introduction

Objectives:

Materials:

Chemicals/ Reagents:

Data and Result:

Solution:

Conclusion
Questions to answer:

1. What component of milk is responsible for its white color?

2. Why is lactose much more soluble in water than in ethanol?

3. What is the purpose of adding 10% acetic acid to the milk colloidal suspension?

4. What is the purpose of adding 1:1 ether-ethanol mixture to casein precipitates?

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