What Colors Your World?
Biotechnology Activities with Food Dyes
Instructors - Bio-Rad Curriculum and Training
Specialists
Sherri Andrews, Ph.D., Eastern US
sherri_andrews@bio-rad.com
Damon Tighe, Western US
damon_tighe@bio-rad.com
Leigh Brown, M.A., Central US
leigh_brown@bio-rad.com
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Separation technology: How do you separate
two or more compounds from each other?
Materials to be Separated
Property Used to
Separate Components
Explanation
Iron filings from other metals
Magnetism
Iron filings will be attracted to a magnet
while other metals will not
Salt from sand
Solubility
Salt will dissolve in water and can be
separated from the sand
Caffeine from coffee beans
Solubility
A solvent is used that selectively
dissolves the caffeine (such as
dichloromethane or ethyl acetate) but
not most of the remaining coffee oils
Ethanol from fermented grain
Boiling point
Ethanol will boil off at a lower
temperature than the other
components of the fermented grain
Wheat from chaff
Density
Wheat kernels are more dense and will
fall to the ground while chaff can be
blow away by the wind
Tea from tea leaves
Size
Filtration can be used to separate the
liquid extract from the large tea leaves
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Combining Science, Engineering and Math to
Develop a Separation Technology
Engineering – Design
parameter based; no single
“correct answer”
Science – Hypothesis
driven; try to find specific
cause/effect
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Analysis
Inexpensive separates
well
Measurements
Best separation material?
Understanding the
chemistry of mobile phaseSalt? Polar? Non-polar?
Chromatography
• Used to separate biomolecules based in their
physical characteristics may include:
•
•
•
•
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size
charge
hydrophobicity
interaction with other molecules
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The System
 All forms of chromatography have a
– Stationary phase (remains stationary and is a solid or a
liquid supported by a solid)
– Mobile phase (a liquid or a gas that travels through the
stationary phase and carries the parts of the mixture)
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Types of Chromatography
 Paper
– Paper serves as the stationary phase.
– The paper strip is placed in a solvent (mobile) phase which
carries the mixture through the paper.
– Parts of the mixture will travel at different rates separating
the parts
 Column
–
–
–
–
–
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The stationary phase is packed into a column
The column is equilibrated (saturated with solvent)
Sample is added to the column
Buffers are added to separate the mixture
Samples are collected in intervals called fractions
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Be an Engineer – Design your own!
 Select materials that you will use to separate your
Kool Aid into the food dyes that make it the color it
appears
 You will do this by
– Paper chromatography
– Column chromatography
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What are some of the design factors we want to
think about?
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Type
Mobile
Phase
Stationary
Phase
Paper
Water
Paper type
Column
1X PBS
HIC Media
Syringe
Alcohol
Cotton
Commercial
Column
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Column
Type
Dye separation from Kool Aid Using Paper
Chromatography
1. Choose your paper type (stationary
phase)
2. Cut it into .75 x 4 inch strips
3. Place 50ul of Kool-Aid at one end using a
DPTP
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4. Place 1 ml of your mobile
phase into a medicine
cup
5. Place the strip Kool-Aid
side down into the mobile
phase
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Building and Using A Column to Separate Your
Kool-Aid
1. Choose your Column Type (HIC, Empty Column,
Syringe)
2. If empty column or syringe add stationary phase by
stuffing it with a cotton ball
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Building and Using A Column to Separate Your
Kool-Aid
3. Choose your mobile phase (water, alcohol, or
1xPBS)
4. Place column in collection tube
5. Add 2 ml of mobile phase to column
6. Let it flow until it no longer drips
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Building and Using A Column to Separate Your
Kool-Aid
7. Move column to next collection
tube
8. Using a DPTP add 1ml of Kool-Aid
to the top of the column
9. Let it flow until it no longer drips (this
is fraction 1)
10. Move to the next column
11. Continue steps 5 and 6 until the
eluate (fluid coming off the
column) is clear
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Compare Results
Which paper, mobile phase worked best for paper
chromatography?
Which column, mobile phase worked best for column
chromatography?
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