HPLC by Mr Bhupendra patidar

1. GRY Institute of Pharmacy, Borawan
High
Performance
Liquid
Chromatography
Prepared By: Bhoopendra Patidar

2. Jens
Martensson
Contents
1. Introduction
2. HPLC Principle
3. Instrumentation
4. Types of HPLC
5. Pump and Sample injection
6. Column
7. Detector
8. Method Development
9. Validation
10. HPLC & UHPLC
11. Advantages
12. Applications 2

3. Jens
Martensson
Introduction
• Chromatography is an analytical method in
which compounds are physically separated
and measured.
• The main purpose of chromatography is to
separate and quantify the target sample.
• The chromatography technique used to
separate mixture of compounds in
pharmaceutical sciences, analytical chemistry
with the purpose of identifying, quantifying
and purifying the individual component of the
mixture.
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4. Jens
Martensson
Principle
• The principle of separation in normal
phase mode and reverse phase mode is
adsorption.
• The component which has more affinity
towards adsorbent travels slower.
• The component which has less affinity
towards statinary phase travels faster.
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5. Jens
Martensson
5
HPLC Component
• Mobile phase reservior
• Degasser
• Pump
• Injector
• Column compartment
• Detector
• Data processor
• Waste
Description
• Stores the mobile phase required for analysis
• Degasses the mobile phases.
• Solvent delivery system, enables the flow of
mobile phases through the system.
• Sample delivery system, introduces to sample
in system
• Control the temperature of column, if
required.
Seperates the analyte components.
• Detects each component in seperated mixture
after it elutes from column.
• Convert the data from detector into results.
• Collection of liquid waste.
Component

6. Jens
Martensson
In Line-Degasser Unit
20YY
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7. Jens
Martensson
Manual Sampler
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8. Jens
Martensson
Auto Sampler
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9. Jens
Martensson
HPLC Instrumentation
9

10. Thanks