PPT ON HPLC CONTAINS INTRODUCTION, INSTRUMENTATION,ADVANTAGES,DISADVANTAGES,APPLICATIONS....

1. HighPerformance Liquid
Chromatography (HPLC)
SHORT NOTE
Saurabh Punia
M-Pharm( 1st year ) 1

2. 
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High Performance
Pressure
Patience
Priced
Precision



Liquid Chromatography
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3. SAURABH PUNIA
 HPLC is a form of liquid chromatography used to separate
compounds that are dissolved in solution.
 HPLC is characterized by the use of high pressure to push a mobile
phase solution through a column of stationary phase allowing
separation of complex mixtures with high resolution.
 HPLC instruments consist of a reservoir of mobile phase, a pump, an
injector, a separation column, and a detector.
 Compounds are separated by injecting a sample mixture onto the
column.
 The different component in the mixture pass through the column at
differentiates due to differences in their partition behavior between the mobile
phase and the stationary phase.
 The mobile phase must be degassed to eliminate the formation of air
bubbles.
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4. PRINCIPLE
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The principle of separation in normal phase mode and reverse
phase mode is adsorption.
When a mixture of components are introduced into a HPLC
column, they travel according to their relative affinities towards
the stationary phase.
The component which has more affinity towards the adsorbent,
travels slower.
The component which has less affinity towards the stationary
phase travels faster. Since no 2 components have the same affinity
towards the stationary phase, the components are separated.
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6. INSTRUMENTATON
:
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1.Solvent delivery
system
2.Pumps
3.Sample injection
system
4.Column
5.Detectors
6.Recorders and
Integrators
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8. HPLC system
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



Solvent Reservoir
Degasser
Solvent Delivery System
(Pump)
Injector
 Column &oven


Detectors
Recorder (Data Collection)
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9. SAURABH PUNIA
HPLCADV
ANTAGES:
 Speed(minutes)
 High resolution
 Sensitivity
 Accuracy
 Automation
HPLC DISADVANTAGES
 Cost
 Complexity 9

10. APPLICATIONS:
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HPLC is one of the most
widely applied analytical separation
techniques.
In Pharmaceutical:
• Tablet dissolution of pharmaceutical
dosages.
• Shelf life determinations of pharmaceutical
products.
• Identification of counterfeit drug products.
• Pharmaceutical quality control.
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11. • Quantitative analysis
The measurement of the amount of a
compound in a sample (concentration);
• Preparation of Pure Compounds
a pure substance can be prepared for later use
(e.g. organic synthesis, clinical studies,
toxicology studies, etc.).
This methodology is called
preparative chromatography
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12. THANKYOU
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