TT Column Chromatography New

7/24/2019 TT Column Chromatography New

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COLUMN

CHROMATOGRAPHY

Group:

Nguyen Hoang Hai AuTran Tuan Ngoc

Luong Nguyen Ngoc Tan

Instructor:

Dr. Nguyen Dieu Lien Hoa


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INTRODUCTION

Column chromatography is generally used as a purification

technique: it isolates desired compounds from a mixture.

This method is very useful when the components of a

mixture have almost the same physical and chemical

properties

This is a solid - liquid technique in which the stationary

phase is a solid and mobile phase is a liquid.

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1. PRINCIPLE

2. COLUMN PACKING TECHNIQUE

3. PREPARING AND LOADING SAMPLE

4. RUNNING THE COLUMN


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The principle of column

chromatography is

based on the

equilibrium of

differential adsorption ofsubstance by the

adsorbent and

intermolecular forces by

the solvent.

1. PRINCIPLE

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1.1 The stationary phase:

A solid adsorbent, is placed in a

vertical glass column.

The stationary phase are usually

finely ground powders or gels or are

microporous for an increased surface.

1. PRINCIPLE

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1.1 The stationary phase:The usual adsorbents employed in

column chromatography are silica,

alumina, calcium carbonate, calcium

phosphate, magnesia, starch, etc.

1. PRINCIPLE

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1.2 The mobile phase:

A liquid mixture, is added to the

top and flows down through the

column by either gravity or

external pressure.

The polarity of the solventwhich is passed through the

column affects the relative

rates at which compounds

move through the column.

1. PRINCIPLE

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Often a series ofincreasingly polar solvent

systems are used to elute

Thin-Layer

Chromatography (TLC) isgenerally used to determine

the system for a column

chromatography separation

with a process of trial and

error.

1. PRINCIPLE

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Proper solvent for each step is chosen so that the retention

factor Rfvalue of the compound of interest is roughly around

0.2 - 0.3 in order to minimize the time and the amount of

eluent to run the chromatography.

1. PRINCIPLE

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There are two kinds of column used for column chromatography:fritted and non-fritted columns.

Fritted column (left) and non-fritted column (right)10

2.1 Preparing the column:


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a) For the fritted columns:

1. Find a clean, empty

column of suitable size.

2. Clamp the column securely

and close the tap or stopcock

(Fig. 2, 2).

3. Add a layer of sand

(approx. 0.5 cm, optional).

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2. COLUMN PACKING TECHNIQUE2.1 Preparing the column:


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b) For the non-fritted columns:

The same as the preparation of fritted

columns but we must add a ball of cotton or

glass wool to plug the bottom of the column.

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2. COLUMN PACKING TECHNIQUE2.1 Preparing the column:


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2.1 Preparing the column:


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Purpose of adding sand: This will give the stationary phase

an even base and prevent concentration and streaking of the

bands as they come off the column and are collected.

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2.1 Preparing the column


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2.2 Filling the Column

Dry-pack method 1:

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There are several methods for filling

columns.


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Dry-pack method 2:



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Slurry Method.

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2. COLUMN PACKING TECHNIQUE2.2 Filling the Column


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Slurry Method: Step C

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2. COLUMN PACKING TECHNIQUE2.2 Filling the Column


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Slurry Method: Step C



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Slurry Method: Step D



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Before After

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Gently using a glass stick to

make the surface of

stationary phase more

evenly.

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3. LOADING THE COLUMN

Solvent:may be a mixture of non-polar solvent and polar

solvent if a sample has many compounds include non-polar

and polar compounds

Dissolve the sample in the minimum amount of solvent (510drops). A slightly more polar solvent than the one you will be

using for the column can be used if this helps with dissolution of

the sample.


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3 LOADING THE COLUMN


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3 LOADING THE COLUMN


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Add a layer of sand(approx. 25 mm) to

protect the surface of

the silica

Finally, add asmuch eluent as

required.

Use a pipette tocarefully add more

eluent

above the top of the

sand


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4. RUNNING THE COLUMN


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http://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_fo

r_the_Lab_Column_Packing.html

http://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Columnchr

om.html Organic chemistry laboratory experiment 1, HCMUS

REFERENCES
http://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_for_the_Lab_Column_Packing.htmlhttp://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_for_the_Lab_Column_Packing.htmlhttp://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_for_the_Lab_Column_Packing.htmlhttp://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_for_the_Lab_Column_Packing.htmlhttp://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Columnchrom.htmlhttp://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Columnchrom.htmlhttp://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Columnchrom.htmlhttp://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Columnchrom.htmlhttp://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Columnchrom.htmlhttp://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_for_the_Lab_Column_Packing.html


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