Centrifugation of Fresh Milk and Acetic Acid

INTRODUCTION
Centrifugation is a process which material suspended in a liquid medium
undergo separation or concentrate due to the effect of gravity. Particles with
different density and masses are settled at different rates in a tube which are
responsive to gravity. Centrifuge is an instrument which can spin carrier vessels at
high rotation speed and very high centrifugal force to allow centrifugation. The
centrifugal force generated is proportional to the rotation rate of the rotor and the
distance between the rotor center and the centrifuge tube. There are three general
classes of centrifuges: low speed about 5000 rpm, high speed machine can up to
about 25000 rpm and ultracentrifuges turn at up to 1000000 rpm. Microcentrifuge is
common in laboratories which generate between 10000 rpm and 13000 rpm and
0.5 or 1.5 ml of disposable plastic tubes are used. Microcentrifuge has only settings
for speed (rpm) but not for the relative centrifuge force (rcf).

OBJECTIVES
1. To separate coagulated milk from mixture of fresh milk and Acetic Acid by
Centrifugation.
2. To determine the relationship of speed of centrifugation with the coagulation
of milk that separates from the mixture.
3. To study the process of centrifugation.

MATERIALS
Centrifuge
Centrifuge tubes (10ml)
Micropipettes
Pipettes
Fresh Milk
50% Acetic Acid

PROCEDURE
1. 10ml of 50% of acetic acid was drawn by pipettes with plastic pipette pump
and released into 50ml Falcon Tube. 5ml of fresh milk was then drawn and
released into a falcon tube with 10ml of acetic acid.
2. The falcon tube was inverted gently for a few times and both solutions are
mixed.
3. The milk sample was centrifuged at 1300 rpm for 10ml minutes, then the
coagulated milk solid was pelleted.

4. 1ml of milk was transferred from the falcon tube to the centrifuge tube. The
centrifuge tube was operated at 5000rpm for 15 minutes.
5. 1ml of milk was transferred from the centrifuge tube to the microcentrifuge
tube. The microcentrifuge tube was operated at 13000 rpm for 15 minutes.
6. Differences between these tubes was observed and results are drawn.

OBSERVATION:
For the first trial, it was observed that within 15 minutes duration having
speed of 1300 rpm the acetic acid was slightly separated from the milk. On second
trial, the acetic acid is highly separated using the speed of 1500 rpm at the same
duration of time. For the last trial, it is proven that the more you increase the speed
of the centrifuge, the more the mixtures became separated with each other.
QUESTIONS
1. What is the driving force in this separation technique?
The driving force in this separation technique is centrifugal force.
Centrifugation is a process which involves the application of the centripetal
force for the sedimentation of heterogeneous mixtures with a centrifuge, and
is used in industrial and laboratory settings. This process is used to separate
two immiscible substances. More-dense components of the mixture migrate
away from the axis of the centrifuge, while less-dense components of the
mixture migrate towards the axis.
2. What purpose does acetic acid serve in this experiment?
The acetic acid was used to lower the pH of the milk and for the protein
(casein) to reach its isoelectric pH. When this occurs, the casein will separate
from the solution.
3. How can we determine that the separation is complete? Relate your answer
to the results obtained at different speeds.
We can determine complete separation by observing. If you increase
speed of centrifugation, the mixtures tend to separate completely with each
other. This can be seen from the experiment performed.

CONCLUSION
Centrifugation is a mechanical process of separating multi-phase mixture via
the use of centrifugal force. The separation of milk from mixture of fresh milk and
acetic acid occurred by the centrifugation process. This is due to the increased
speed of centrifugation. Therefore, the relationship between speed of centrifugation
to separation of mixtures is directly proportional. To increase speed is to give better
chance for mixture to separate.