student

1. PRESENTED BY
HIBA.C.H
1ST YEAR M-PHARM
DEPT. OF PHARMACEUTICS

2. • Chromatography
Greek :chromatos -- color ,
"graphein" -- to write.

3. Definition
The separation of a mixture by distribution of its
components between a mobile and stationary
phase over time
OR
The separation of mxt of components into
individual components through equilibrium
distribution b/w two phases

4. • Mobile phase - phase that moves through
chromatograph
• Stationary phase - phase that is stationary in
chromatograph

6. Real-life examples of uses for chromatography:
• Pharmaceutical Company – determine amount of each
chemical found in new product
• Hospital – detect blood or alcohol levels in a patient’s
blood stream
• Law Enforcement – to compare a sample found at a
crime scene to samples from suspects
• Environmental Agency – determine the level of
pollutants in the water supply
• Manufacturing Plant – to purify a chemical
needed to make a product

7. There are two classification schemes:
–Mobile phase
–Attractive forces

8. Mobile Phase
• Gas (GC)
• Water (LC)
• Organic solvent (LC)
• Supercritical fluid (SCFC)

9. Classification based on Mobile Phase
Gas
Chromatography
Gas - solid
Gas - liquid

10. Liquid chromatography
(LC)
Liquid- solid
Column
(gravity
flow)
High
performanc
e(pressure
flow)
Thin
layer
(adsorpt
ion)
Liquid -liquid
Paper –
partition
Column
partition

11. 1. Adsorption
2. Ion Exchange
3. Partition
4. Size Exclusion

12. Separation based on their adsorption onto the
surface of solid (stationary phase).
 the compound which has more affinity
towards stationary phase travels slower.
Ex; Column chromatography (CC) , TLC,
HPLC

13.  solute are separated based on their partition between a
liquid mobile phase and a liquid stationary phase
coated on a solid support.
 Component which is more soluble in mobile phase
travels faster & more soluble in stationary phase
travel slower.
Ex : liquid chromatography, Paper Chromatography,
column partition chromatography

14.  Use ionic stationary phase
– ions separated on the basis of their tendency to
displace counter ions adsorbed on stationary phase
(Depends on charge, hydration, “solubility”…)
 Anionic stationary phases: used for cation separation
 Cationic stationary phases : for anion separation
 for ionic compounds
 - Ex : CC , HPLC

15. Separation is a result of “trapping” of molecules
in the pores of the packing material
• Very large molecules can’t get into the pores –
unretained
• Very small molecules get hung up in to pores for a
long time - most retained – longest retention time
• stationary phase is a porous matrix
• Ex: CC, HPLC

18. Types of Chromatography
• Liquid Chromatography – separates liquid
samples with a liquid solvent (mobile
phase) and a column composed of solid
beads (stationary phase)
• Gas Chromatography – separates vaporized
samples with a carrier gas (mobile phase) and
a column composed of a liquid or of
solid beads (stationary phase)

19. Types of Chromatography
• Paper Chromatography – separates dried
liquid samples with a liquid solvent (mobile
phase) and a paper strip (stationary phase)
• Thin-Layer Chromatography – separates
dried liquid samples with a liquid solvent
(mobile phase) and a glass plate covered
with a thin layer of alumina or silica gel
(stationary phase)

20. STATIONARY PHASE
Type of chromatography Material
Paper chromatography Filter paper, cellulose
Thin Layer Chromatography Silica gel, alumina,
polyamide
Gas chromatography
(GC)
Squalene, apezion, carbowax
M
High Performance Liquid
Chromatography
C-8, C-18, Licosorb, Silicone

21. MOBILE PHASE
Type of chromatography Solvent
Paper chromatography Air, alcohol
Thin Layer Chromatography Hexane, ether petroleum, alcohol.
Gas chromatography
(GC)
He, Ar, N2
High Performance Liquid
Chromatography
Cyclohexane, n-hexane, carbon
tetrachloride, ethanol, methanol,
air