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Lecture 7. Chromatography Intro!. Solid Phase Extraction. Load (sample). Eluent (solvent). Compounds of interest stick to sorbent. Compounds of interest released from sorbent. Waste. Extract. Chromatography. Mobile phase. Mobile phase. Load Sample (injection). - PowerPoint PPT Presentation
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Lecture 7
Chromatography Intro!
Compounds of interest stick
to sorbent
Solid Phase ExtractionLoad
(sample)
Compounds of interest
released from sorbent
Waste
Eluent(solvent)
Extract
Compounds of interest in a
plug
Chromatography
LoadSample
(injection)
Differential interaction with
stationary phase separates compounds in
time and space.
Detector
Mobile phase
Detector
Mobile phase
ChromatographyDefinition
Separation of components in a mixture based on the differential movement through a porous media under the influence of a mobile phase
First Chromatography Column
toluene
sample
What happens in a chromatographic column?
Gas Chromatography(GC)
Liquid Chromatography(LC)
Distillation Liquid-liquidextraction
Theoretical Plate
Silica support
Stationary phase
GC - gas chromatographyopen tubular columns
Gas (mobile phase)
Theoretical PlateLC - liquid chromatography
packed columns
Silica beads
Stationary phase
liquid (mobile phase)
Number of Theoretical Plates
N = L H
Length of the column
Height equivalent of a theoretical plate
Retention Factor
k’ = tR-tM
tM
= VR-VM
VM
1 < k’ < 10
Describes the strength of the interaction of a compound with the stationary phase
Compounds of interest stick
to sorbent
Solid Phase ExtractionLoad
(sample)
Compounds of interest
released from sorbent
Waste
Eluent(solvent)
Extract
k’ > 1000 k’ < 0.001
Compounds of interest in a
plug
Chromatography
LoadSample
(injection)
Differential interaction with
stationary phase separates compounds in
time and space.
Detector
Mobile phase
Detector
Mobile phase
1 < k’ < 10
Retention Factor (k’)Describes the strength of the interaction of a
compound with the stationary phase
k’ = tR-tM
tM=
VR-VM
VM
1 < k’ < 10
k’ = tR-tM
tM=
VR-VM
VM
1 < k’ < 10
Retention Factor (k’)
Effect of k’ on Selectivity and Resolution in LC
Dong, M. Today’s Chemist at Work. 2000, 9(2), 46-48.
SelectivityDescribes differential mobility of 2 compounds
= k’2
k’1
= K2
K1
ResolutionDescribes how well 2 compounds are separated
Rs = 2(tR2-tR1)
tw1+tw2
ResolutionDescribes how well 2 compounds are separated
Rs = 14
N1/2 (-1)k’
1+k’( )efficiency selectivity
retention
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