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Spinach TLC
11
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Types of Chromatography
How does it Get Separated?
Paper Chromatography*
Thin Layer Chromatography (TLC)*
Gas Chromatography
Liquid Chromatography
Ion Exchange Chromatography
Affinity Chromatography etc…
Different compounds in the mixture travel through the phases at different rates
Compounds with a smaller affinity for the stationary phase spend travel faster with the mobile phase.
*Quick and inexpensive techniques to qualitatively find out how many components are in a mixture
What does it need to separate?
Why it is Important?
- Stationary Phase – TLC Plate
(Silica Gel) ->Polar
Chemistry
Biology
Biotechnology
Pharmaceutical Industry
Environmental
Food
Fragrances
Forensics
- Mobile Phase –
70:30 Hexane: Acetone v/v -> Nonpolar
Thin Layer Chromatography of Spinach Pigments
Chromatography
Isolation of Pigment from Leaves
- Grind 1g of spinach leaves with pestle and mortar with 2mL of acetone until consistency is creamy
A technique used to separate and analyze mixtures
Qualitative and quantitative analysis
- Add additional 1mL of acetone to spinach and transfer contents to glass vial
- Allow vial to set to achieve separation of aqueous and organic layer
Spinach Chromatography
Science Expo
11 October 2012
- Remove aqueous layer and add NaSO4 to remove traces of water
- Filter organic layer using a cotton plug and pipette
- Organic layer is ready to be analyzed using TLC
What makes up Matter?
Thin Layer Chromatography of Pigments
- Prepare a chamber with mobile phase,7mL of hexane acetone, transfer to 150mL beaker.
- Dip filter paper in mobile phase then hang on the side of the beaker, seal beaker with foil
- Make a line 1 cm from bottom and 3mL of TLC plate, origin line
- Spot organic layer on origin line with a microcapillary tube, let dry, do this 3 times
- Place upright in chamber and seal with foil
- Observe as you see pigmentation separate while solvent moves up TLC plate, end once 1cm from top
- Remove plate and immediately trace solvent line
What is Chemistry?
How do we identify the compounds in the Mixture?
Rf = Retention Factor
- Chemistry is a study of the composition, structure, and properties of matter and changes that occur.
- Chemists study both qualitative and quantitative properties of matter.