Thin Layer Chromatography (TLC)

```Thin Layer
Chromatography (TLC)
Separatory Techniques: Chromatography
What Is Chromatography?
▪ Chromatography is separating molecules
TLC System: 3 Components
▪ Stationary Phase (Chromatography Plate)
▪ Mobile Phase: Solvent
▪ Analyte: Sample blotted on TLC plate
Stationary and Mobile Phases
▪ Stationary Phase: A
glass/metal/polymer plate most
commonly coated with a polar silica
dioxide or aluminum dioxide matrix.
▪ Mobile Phase: A low polarity solvent
or mix of two or more low polarity
solvents. Ex. Hexanol or 1:1 Mix of
Hexanol and THF
Seperation: Interaction of the analytes, solvent and stationary matrix
1.Prepare
TLC Place
Draw, Blot
and Place
in Vessel
2.Seperate
allow separation
to occur
3. Results
allow separation
to occur
Separation: A polar Interaction
Before Separation (0 min)
After Separation (10 min)
Rf: Retention Factor
Rf
Rf A = 8 cm / 10 cm = 0.8
=
Distance solute traveled
----------------------------------Distance solvent front traveled
Rf B = 3 cm / 10 cm = 0.3
Rf C = 2 cm / 10 cm = 0.2
Apply Your
Knowledge
A TLC plate is blotted with 3 samples
and allowed to separate for 10
minutes. In the end the calculated
retention factors are:
0.65
0.30
0.95
Which molecules has the Rf of 0.30?
• Chromatographic method using a polar stationary phase on
a plate and a less polar mobile phase
• Analyte is carried up the stationary matrix by the mobile
phase
Rf = (Distance analyte traveled / distance solvent front)
• The more polar a substance is the less time it will be in the
mobile phase = Travels less, has an Rf closer to 0
• The less polar a sample is the more time it will be in the
mobile phase = Travels more, has an Rf closer to 1
Thin Layer
Chromatography
An Overview
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