DEPARTMENT OF TECHNICAL EDUCATION

ANDHRA PRADESH
Name : A.Vinay Kumar
Designation : H.O. D
Branch : Applied Electronics and Instrumentation
Institute : Q.Q.GPT, Hyderabad
Year/Semester : VI SEM
Subject : Analytical Instrumentation
Code No : AEI-603
Major Topic : Mass spectroscopy & Chromatography
Duration : 100 Mts
Sub Topic : Types, applications and detectors of
Liquid chromatography
Teaching Aids : PPT’s, Diagrams
AEI603.39 to 40 1
 Objectives

On the completion of this topic you would

able to know about Liquid Chromatography

• Types

• Applications

• Detectors

• Limitations

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 Recap

In last class we have studied about Liquid

chromatograph

• Operation

• Principle

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 Types of HPLC

1. Normal Phase Chromatography (NPC)

2. Revesed Phase Chromatography (RPC)

3. Size Exclusion Chromatography (SEC)

4. Ion Exchange Chromatography (IEC)

5. Bio -affinity Chromatography (BAC)

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1. Normal phase Chromatography (NPC)

• This is the first kind of HPLC chemistry

• It separates analytes based on the polarity

• It uses a polar stationary phase and non-polar mobile

phase

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2.Revesed Phase Chromatography (RPC)

• It consists of a non-polar stationary phase, e.g.-silica

• An aqueous, moderately polar mobile phase

• It operates on the principle of hydrophobic interactions

AEI603.39 to 40 6
3.Size exclusion chromatography

• It is also known as ‘gel permeation chromatography or

gel filtration chromatography’

• It separates particles on the basis of size

• It is a low resolution chromatography

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4. Ion exchange chromatography

• Retention is based on the attraction between solute ions

and charged sites bound to the stationary phase

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5. Bioaffinity chromatography

c. It relies on the property of the biologically active

substances to form

i) Stable

ii) Specific

iii) Reversible Complexes

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Liquid chromatography

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Liquid chromatography

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 Applications

• In pharmaceutical industry to qualify drugs before they

release

• Molecular weight comparison

• Molecular weight determination

• Water purification

• Pre-concentration of trace components

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 Detectors

• UV-Visible spectrometric detectors

• Fluorescence detector

• Refractive index detector

• Adsorption detector

• Electrical conductive detector

• Thermal detector

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UV-Visible spectrometric detector

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 Sterdation Cuvette

Mounting cage
Sample cuvtte
2Π steridian
Interceptor optic

Chamber bar

Emitted light
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 Conductivity detector

Chromatographic column

Glass
conductivity cell

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 Limitations

• Maintenance of the solvent flow accurately

• Problem in controlling solvent flow arise

• The column must be tightly packed

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 Summary

In this class we have learnt about Liquid

Chromatography

• Types

• Applications

• Limitations

• Detectors

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 Quiz

1. Which of the following is detector of LC

a) UV-Visible spectrometric detectors

b) Fluorescence detector

c) Refractive index detector

d) All the above

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 Frequently Asked Questions

1.List the applications and detectors of Liquid

chromatography?

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