an introduction to chromatography
DESCRIPTION
An Introduction to Chromatography. What is chromatography? The separation of a mixture by distribution of its components between a mobile and stationary phase over time mobile phase = solvent or gas stationary phase = column packing material. Purpose of Chromatography. - PowerPoint PPT PresentationTRANSCRIPT
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An Introduction to Chromatography
• What is chromatography?
• The separation of a mixture by distribution of its components between a mobile and stationary phase over time– mobile phase = solvent or gas– stationary phase = column packing material
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Purpose of Chromatography
• AnalyticalAnalytical - determine chemical composition of a sample
• PreparativePreparative - purify and collect one or more components of a sample
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Classification of Column Chromatographic MethodsGeneral Classification Specific Method Stationary Phase Type of Equilibrium
Liquid-liquid orpartition
Liquid adsorbed on solid Partition betweenimmiscible liquids
Liquid bondedphase
Organic liquids bonded on asolid surface
Partition between liquidand bonded phase
Liquid- solid oradsorption
Solid Adsorption
Ion exchange Ion exchange resin Ion exchange
Liquid chromatography(LC)(Mobile phase: liquid)
Size exclusion Liquid in interstices of apolymeric solid
Partition/sieving
Gas-liquid Liquid adsorbed on solid Partition between gas andsolid
Gas-bonded phase Organic species bonded to asolid surface
Partition between gas andbonded phase
Gas Chromatography(GC)Mobile phase: gas
Gas-solid Solid Adsorption
Two-stagepressure regulator
Carrier gas supply
Flow controller
Rotameter
Flow splitter Syringe
Injector
Detector
Temperature controlled
zonesColumn
Schematic of a Gas Chromatograph
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Carrier Gas Supply
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300Vdc
Injector and Detector
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300Vdc
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Injection types
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A Capillary Column for GLC
Fused silica tubingdc 0.3 mm
df 1 mL = 15 - 60 m
Polydimethylsiloxane (silicone)• Good for retaining and separating
nonpolar solutes by boiling point
A thin layer of nonvolatile stationary phase is coating on the inner wall of the
tubing (WCOT)
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Some Common Stationary Phases for Gas-Liquid Chromatography Stationary phase Common trade
name Maximum temperature
Common Applications
Polydimethyl siloxane OV-1,SE-30 350 General purpose nonpolar phase; hydrocarbons; polynuclear aromatics; drugs; PCB”s
Poly(phenylmethyl dimethyl) siloxane (10% phenyl)
OV-3, SE-32 350 Fatty acid methyl esters; alkloids; drugs; halogenated compounds
Poly(phenylmethyl) siloxane (50% phenyl)
OV-17 250 Drugs; steroids; pesticides; glycols
Poly(trifluoropropyldimethyl) siloxane
OV-210 200 Chlorinated aromatics; nitroaromatids; alkyl substituted benzenes
Polyethylene glycol Carbowax 20M 250 Fatty acids; alcohols; ethers; essential oils; glycols
Poly( dicyanoallyldimethyl) siloxane
OV-275 240 Poly unsaturated fayyt acids; rosin acids; free acids; alcohols
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SiO
OH
SiO
OH
SiO
OH
SiO
OH
Si OH + Si ClCl
CH3
CH3
Si O Si Cl
CH3
CH3
Si O Si Cl +
CH3
CH3
CH3OH Si O Si OCH3
CH3
CH3
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12
3
4
5 6 78
9
87
65
5
43
21
0 10 20 30
Isothermal at 45oC
0 10 20 30
Isothermal at 145oC
0 10 20 3030 60 90 120 150 180
TimeTemperature(oC)
Programmed from 30o to
180oC
Effect of temperature on gas chromatograms
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Capillary GC-MS
Fused silica Column
Transfer Line
Quadruple MSGC Data system
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Sample
Carrier
Packed column GC-MS Interface
To MS
Skimmer
To vacuum pump
Nozzel
From GC
Jet separator
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Gaseousion sources
Mass analyzer
Ion transducer
Signalprocessor
Ion inlet
Vacuum pump
10-5 to 10-8 torr
Components of a Mass Spectrometer
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Sample inlet
• batch inlet
• direct probe inlet
• chromatographic inlet
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Vacuum pump
Heating coil
Ionization chamber
Direct insertion probe
A sample probe for inserting a sample directly into an ion source
Direct probe
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Molecular leak
Repeller
Electron beam
HeaterShield
Filament
1st accelerating slit
Focusing slit
2nd accelerating slit
Anode
To mass analyzer
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Ions with unstabletrajectories
Ions with stabletrajectories
Ion source
Dc and ac oltages
Quadruple Mass Spectrometer
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Continuous Dynode Electron Multiplier
Resistive conductivesurface
GND
2kV
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