INTRODUCTION OF CHROMATOGRAPHY

1. BY: CHAURASIYA RAUNAK KUMAR
1ST M PHARMACY
DEPARTMENT OF PHARMACOLOGY
MALLIGE COLLEGE OF PHARMACY
TO : Mrs T.MENAKA
ASSISTANT PROFESSOR

2. CONTENTS :
Definition
Principles
Types of chromatography
Applications of chromatography

3. DEFINITION :
 Derived from two greek words , chroma –colour and graphein– to write
Chromatography is the analytical technique used for separating a mixture of
component into individual components using a stationary and mobile phase .

4. TYPES OF CHROMATOGRAPHY :
Based upon the nature of stationary and mobile phase :
1. Gas – Solid chromatography .
2. Gas –Liquid chromatography .
3. Solid – Liquid chromatography .
[column chromatography ,thin layer chromatography ,HPLC]
4. Liquid – Liquid chromatography .
[paper partition chromatography , column partition chromatography ]

5. Based on the principle of separation :
1. Adsorption chromatography .
2. Partition chromatography .
Other types of chromatography :
1. Ion exchange chromatography .
2. Gel permeation chromatography .
3.chiral chromatography .

6. Based on modes of chromatography :
based on polarity of stationary phase and mobile phase .
1. Normal phase chromatography .
2. Reverse phase chromatography.
NORMAL PHASE REVERSE PHASE
Stationary phase Polar Non polar
Mobile phase Non polar Polar
Comp eluted first and
retained less
Non polar Polar
Comp eluted last and
retained more
Polar Non polar
Ex of stationary phase Silica gel C8 , C4 .

7. PRINCIPLES OF CHROMATOGRAPHY:
1 . ADSORPTION CHROMATOGRAPHY :
When a mixture of compounds (adsorbate) dissolved in the mobile phase (eluent) moves
through a column of stationary phase (adsorbent)they travel according to the relative
affinities towards stationary phase and mobile phase .
Examples :gas –solid chromatography
thin layer chromatography
column chromatography
HPLC .

8. ADSORPTION CHROMATOGRAPHY :

9. 2. Partition chromatography :
 When two immiscible liquids are present ,a mixture of solutes will be distributed
according to their partition co- efficients.
When a mixture of compounds are dissolved in the mobile phase and passed
through a column of liquid stationary phase ,the components are separated based on their
affinities towards stationary and mobile phase .
Examples: Gas- liquid chromatography
Paper partition chromatography
Column partition chromatography .

10. PARTITION CHROMATOGRAPHY :

11. 3. Ion exchange chromatography :
 In this type ,an ion exchange resin is used .Reversible exchange of ions takes place
between similar charged ions and that of ion exchange resin .
A cation exchange resin is used for separation of cation and an anion exchange resins
used to separate a mixture of anions.

12. ION EXCHANGE CHROMATOGRAPHY:

13. 4.Gel permeation chromatography:
 A gel is used to separate the components of a mixture according to their molecular sizes .
 Different gels are used for different molecular weight ranges .
 Solvent used can be aqueous or non aqueous .
 Stationary phase is a porous matrix .
 Mobile phase used may be organic solvents or aqueous buffers .
 The mechanism involved in separation is because of steric and diffusion effects in the
pores of different gels .

14. GEL PERMIATION CHROMATOGRAPHY:

15. 5. Chiral chromatography :
 In this type of chromatography ,optical isomers ( levo and dextro )can be separated by
using chiral stationary phases .
 Enantiomers are separated based on the number and type of each interaction that occurs
during their exposure to chiral stationary phase.

16. CHIRAL CHROMATOGRAPHY:

17. APPLICATION OF
CHROMATOGRAPHY:
Used in pharmaceutical industry –
To identify purity of drugs
identify quantity of impurities.
Food and beverage industry—
to identify pesticides content in beverages and heavy metal content in water or
food stuffs .
To separate chiral compounds –
ex: separation of dextro and levo ketoprofen.

18. Cont……
Complex mixtures are separated and analysed—
separation and identification of carbohydrates , amino acids , fatty acids and
other natural substances .
Identification of drugs –
ex: chlorpromazine
stationary phase :silica gel G
mobile phase :ether :ethyl acetate : ammonia
detection : uv at 254 nm.
In forensic science –
used widely in testing crime scene like analysing blood and hair samples .

19. References :
 Text book of pharmaceutical analysis – Dr . S . Ravi Sankar .
 https ://www.slideshare.net>namarta28.
 Instrumental method of chemical analysis –Gurdeep R. Chatwal.

20. THANK YOU