2. THE CHROMATOGRAPHIC PPRROOCCEESSSS -- PPAARRTTIITTIIOONNIINNGG
(gas or liquid)
MMOOBBIILLEE PPHHAASSEE
SSTTAATTIIOONNAARRYY PPHHAASSEE
Sample
out
Sample
in
(solid or heavy liquid coated onto a solid or support system)
4. Common Types of Chromatography
Tswett’s technique is based on Liquid Chromatography. There
are now several common chromatographic methods. These
include:
1. Paper Chromatography
2. Thin Layer Chromatography (TLC)
3. Liquid Chromatography (LC)
4. High Pressure Liquid Chromatography (HPLC)
5. Ion Chromatography
6. Gas Chromatography (GC)
6. EEqquuaattiioonn ffoorr PPaarrttiittiioonn RRaattiioo
A A
A
A
A
A
A
A
A
A
A
A
K = distribution
coefficient or
partition ratio
K =
CS
CM
Where CS is the molar
concentration of the
solute in the stationary
phase and CM is the
molar concentration in
the mobile phase.
Cross Section of Equilibrium in a column.
“A” are adsorbed to the stationary phase.
“A” are traveling in the mobile phase.
16. CARRIER GAS
» Hydrogen
• Better thermal conductivity
• Disadvantage: it reacts with unsaturated compounds
& inflammable
» Helium
• Excellent thermal conductivity
• It is expensive
» Nitrogen
• Reduced sensitivity
• It is inexpensive
17. RReeqquuiirreemmeennttss ooff aa ccaarrrriieerr ggaass
Inertness
Suitable for the detector
High purity
Easily available
Cheap
Should not cause the risk of fire
Should give best column performance
18. AADDVVAANNTTAAGGEESS OOFF GG..CC
1. Very high resolution power, complex mixtures can be
resolved into its components by this method.
2. Very high sensitivity with TCD, detect down to 100
ppm
3. It is a micro method, small sample size is required
4. Fast analysis is possible, gas as moving phase-rapid
equilibrium
5. Relatively good precision & accuracy
6. Qualitative & quantitative analysis is possible
19. Disadvantages of G.C.
1. The samples analysed are limited to those that
are volatile or can be made volatile.
2. The samples must be thermally stable to
prevent degradation when heated.
3. Cannot be used to prepare samples for further
analysis once separated.
4. Problems can be encountered when injecting
the sample
20. Applications of G.C
• G.C is capable of separating, detecting & partially
characterizing the organic compounds , particularly
when present in small quantities.
1, Qualitative analysis
Rt & RV are used for the identification & separation
2, Checking the purity of a compound
Compare the chromatogram of the std. & that of the
sample
21. Applications of G.C
3, Quantitative analysis
It is necessary to measure the peak area or peak
height of each component
4, Used for analysis of drugs & their metabolites.