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Dive into the world of food dye separation through chromatography processes. Discover various techniques, materials, and factors affecting separation efficiency. Practice hands-on experiments to unveil the mysteries of color separation.
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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
Separation technology: How do you separate two or more compounds from each other?
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 Best separation material? Measurements Analysis Understanding the chemistry of mobile phase- Salt? Polar? Non-polar? Inexpensive separates well
Chromatography • Used to separate biomolecules based in their physical characteristics may include: • size • charge • hydrophobicity • interaction with other molecules
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)
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 • 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
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
Dye separation from Kool Aid Using Paper Chromatography • Choose your paper type (stationary phase) • Cut it into .75 x 4 inch strips • Place 50ul of Kool-Aid at one end using a DPTP
Place 1 ml of your mobile phase into a medicine cup • Place the strip Kool-Aid side down into the mobile phase
Building and Using A Column to Separate Your Kool-Aid Choose your Column Type (HIC, Empty Column, Syringe) If empty column or syringe add stationary phase by stuffing it with a cotton ball
Building and Using A Column to Separate Your Kool-Aid • Choose your mobile phase (water, alcohol, or 1xPBS) • Place column in collection tube • Add 2 ml of mobile phase to column • Let it flow until it no longer drips
Building and Using A Column to Separate Your Kool-Aid • Move column to next collection tube • Using a DPTP add 1ml of Kool-Aid to the top of the column • Let it flow until it no longer drips (this is fraction 1) • Move to the next column • Continue steps 5 and 6 until the eluate (fluid coming off the column) is clear
Compare Results Which paper, mobile phase worked best for paper chromatography? Which column, mobile phase worked best for column chromatography?