3.16 chromatography Flashcards
all forms of chromatography involve the distribution of the components of a mixture between which 2 phases
a stationary phase and a moving phase
phase
a state such as solid, liquid or gas
what does the mobile phase pass over
the stationary phase
what doesn’t the stationary phase do
move
adsoprtion
the process by which a sold holds molecules onto its surface
what does TLC stand for
thin layer chromatography
what does cc stand for
column chromatography
what does GC stand for
gas chromatography
TLC: stationary phase
piece of card/filter paper
TLC plate- silica/alumina coating
glass
TLC: mobile phase
various solvents used
CC: stationary phase
glass column packed with a solid
CC: mobile phase
a liquid solvent (eluent) moves down the column
GC: stationary phase
solid/solid coated with lqiuid packed into a capillary tube
TLC: mobile phase
unreactive carrier gas (eg He/N2) used under pressure at high temp
separation of components: how does separation occur if the stationary phase is a solid
adsorption
separation of components: what is the correlation between strength of adsorption and speed of component moving through mobile phase
the stronger the adsorption to the stationary solid phase, the slower the component moves through the mobile phase
separation of components: how does separation occur if the stationary phase is a liquid
by relative solubility
separation of components: what is the correlation between solubility of liquid stationary phase and speed of component moving through mobile phase
the greater the solubility in the stationary liquid phase, the slower the component moves through the mobile phase
separation of components: what is the rate of movement of a component recorded as and what can this be used to idetify
recorded as Rf value/retention time
can be used to identify a component
separation of components: what properties is the strength of adsorption and relative solubility of a molecule affected by
-charge
-polarity
stereoisomerism
separation of components: what does separation depend of the balance between
solubility in the moving phase and retention in the stationary phase
TLC: uses
separation- identifying components in mixtures
TLC: moving phase
solvent
TLC: stationary phase
TLC plate
TLC experimental details: dissolve a small sample of mixture in
solvent
TLC experimental details: draw a … a short distance from the bottom of the TLC plate
pencil line
TLC experimental details: place a spot of… on the pencil line and allow to dry
component
TLC experimental details: place the TLC plate in a beaker containing a small amount of
the solvent
TLC experimental details: cover beaker with
a lid
TLC experimental details: allow solvent to rise up tlc plate and stop before
it reaches the top
TLC experimental details: mark on the … in … once the solvent has risen almost to the top of the TLC plate
solvent front in pencil
TLC experimental details: what can be used to make the compounds visible
locating agent
TLC: locating agents
uv light, ninhydrin, I2 crystals, MnO4
TLC: advantages compared to paper chromatography
- faster
- requires smaller amounts of mixtures
- better resolution
TLC: limitations
- similar compounds have similar Rf values
- unknown compounds have no Rf values for reference
- can be difficult to find a solvent to separate all components in a mixture
Rf
distance moved by spot/distance moved by solvent
Rf values can only be between
0 and 1
CC: uses
separating large amounts of mixtures
CC: moving phase
liquid solvent which passes down and out of column (eluent)
CC: stationary phase
solid beads
CC experimental details: fill a glass tube with
stationary phase, held in place by a filter or mineral wool plug
CC experimental details: cover all powder in
solvent
CC experimental details: dissolve mixture to be sampled in
minimum amount of solvent
CC experimental details: place mixture on top of
solid phase
CC experimental details: run mixture through column by
opening tap and adding solvent at the top
CC experimental details: time taken for each component to reach end of column recorded, this is known as
retention time
CC: advantages
-used for larger amounts of sample
GC: what kind of method is GC
a very sensitive quantitative method
GC: uses
to test a wide range of compounds
GC: moving phase
inert carrier gas at high temp and pressure
GC: stationary phase
thin layer of solid/liquid- both in capillary tube
GC experimental details: mixture injected into gas chromatograph where it
vapourises
GC experimental details: what flushes mixture through the column
the carrier gas (moving phase)
GC experimental details: why do the components slow down
as they interact with the stationary phase
GC experimental details: each component leaves the column at a different time and is
detected
retention time
the time taken for a component to pass from the column inlet to the detector
correlation between retention time and components solubility in stationary phase
the longer the retention time, the greater the components solubility in the stationary phase
why can retention times be theoretically used for identification
different compounds have different retention time values
gas chromatogram: number of peaks=
number of components in the mixture
gas chromatogram: area under each peak proportional to
amount of a component in the mixture
limitations of GC:
- many 1000s compounds have same retention time and peak shape
- peaks for components present in high conc can hide smaller peaks with same retention time
- unknowns have 0 reference
what does GCMS stand for
gas chromatography mass spectrometry
what does combining gas chromatography with mass spectrometry give
a far more powerful tool than gas chromatography alone
what is gas chromatography not good at despite being used to separate components
identifying structure
GCMS: what is mass spectrometry used to do
identify the separated components
GCMS: first stage
GC first used to separate components in the mixture
GCMS: second stage
each separated compoentns directed to MS in turn
GCMS: third stage
each mass spectrum can be analysed/compared with spectral database, so enabling compound to be identified
GCMS: fourth stage
quantity of each component can also be determined
what is a mass specturm
a plot of relative abundance against mass to charge ratio
what does a mass spectrum consist of
a series of peaks at different masses corresponding to the mass of the whole molecule and the masses of the fragment ions
what will the peak with the highest m/z ratio in a mass spectrum be down to
the molecular ion
what does fragmentation provide in mass spectrums
structural detail
uses of GCMS
- forensic and drug analysis
- environmental analysis
- airport security
- space probes