what colors your world?

What Colors Your World? Biotechnology Activities with Food Dyes

Biotechnology Explorer™ | explorer.bio-rad.com2

Instructors - Bio-Rad Curriculum and Training Specialists

Sherri Andrews, Ph.D., Eastern US

[email protected]

Damon Tighe, Western US

[email protected]

Leigh Brown, M.A., Central US

[email protected]


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


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

Inexpensive separates well

Understanding the chemistry of mobile phase- Salt? Polar? Non-polar?

Best separation material?

Mea

sure

men

ts

An

alys

is


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


What are some of the design factors we want to think about?

Type Mobile Phase

Stationary Phase

Column Type

Paper Water Paper type

Column 1X PBS HIC Media Syringe

Alcohol Cotton Commercial Column


Dye separation from Kool Aid Using Paper Chromatography

1. Choose your paper type (stationary phase)2. Cut it into .75 x 4 inch strips3. Place 50ul of Kool-Aid at one end using a

DPTP


4. Place 1 ml of your mobile phase into a medicine cup

5. Place the strip Kool-Aid side down into the mobile phase


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

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



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



Compare Results

Which paper, mobile phase worked best for paper chromatography?

Which column, mobile phase worked best for column chromatography?

what colors your world?

Documents

mixturebiotechnology

astationary phase

solvent mobile phase

separate biomolecules

paper type stationary

dye separation

paper chromatographychoose

paper strip