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Protein Purification By: Presentation On

The document provides an overview of protein purification using high performance liquid chromatography (HPLC). [1] HPLC is a technique used to separate, identify, detect, and quantify components in a solution under high-pressure conditions that provide enhanced separations over shorter periods. [2] The principle of HPLC involves using a high-pressure pump attached to a column that is pressurized, with either non-polar or polar mobile and stationary phases used depending on the type of HPLC. [3] HPLC has applications in the pharmaceutical industry for drug stability and quantification, analysis of environmental contaminants, and forensic testing.

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Nafiz Fuad
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30 views

Protein Purification By: Presentation On

The document provides an overview of protein purification using high performance liquid chromatography (HPLC). [1] HPLC is a technique used to separate, identify, detect, and quantify components in a solution under high-pressure conditions that provide enhanced separations over shorter periods. [2] The principle of HPLC involves using a high-pressure pump attached to a column that is pressurized, with either non-polar or polar mobile and stationary phases used depending on the type of HPLC. [3] HPLC has applications in the pharmaceutical industry for drug stability and quantification, analysis of environmental contaminants, and forensic testing.

Uploaded by

Nafiz Fuad
Copyright
© © All Rights Reserved
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Presentation On

PROTEIN
PURIFICATION BY
HPLC PRESENTED BY : NAFIS FUAD
ID No. : B180607029
COURSE NAME : FUNDAMENTALS OF GENETIC
ENGINEERING AND BIOTECHNOLOGY
COURSE CODE : BMB3102
GROUP : E
DATE : 29th AUGUST, 2022
CONTENTS

• What is HPLC ? (P: 01)


• Discovery of HPLC (P: 01)
• Principle of HPLC (P: 01)
• Mechanism and screening (P: 02)
• Advantages and disadvantages of HPLC (P: 03)
• Applications of HPLC (P : 03)
P: 01
What is HPLC?
HPLC (High Performance Liquid Chromatography or High Pressure Liquid Chromatography) is
a technique used to separate, identify, detection and quantify each component in a solution
under conditions which provide for enhanced separations during shorter periods of time.
This is both quantitative and qualitative biochemical assay.

Principle of HPLC FUN FACTS


• High pressure pump is attached to the column which is
pressurized by approximately 40 MP. Martin and Synge
• For normal phase HPLC Non-Polar Solvent and Polar Silica established HPLC
technique while they
beads and for reverse phase HPLC Polar mobile Phase and were working at the
Non-polar stationary phase is used. Wool Industries
• Sample molecules passes (Here Peptides) travel towards the Research Association
in England
detector and these molecules are separated by their polarity.
• Different types of detectors are used, such as : UV detector,
IR detector, mass spectroscopy, refractive index detector etc.
P: 02
How Does It Work?

What’s The Ultimate Scenario?

How To Interpret Data?


P: 03
Advantages Of HPLC
• High resolving power.
• High speed separation. Disadvantages Of HPLC
• Automated sampler.
• More reproducible. • Both the setup and columns are expensive.
• Adaptable for industry level usage. • Tedious to detect co-elusion.
• Column can be reused. • Complex to operate.

Applications Of HPLC
• Pharmaceutical industry:
1. To control drug stability
2. Quantity of drug determination from biological fluids. Ex : Blood glucose level.
• Analysis of natural contamination. Such as Phenol and Mercury from sea water.
• Forensic test :
1. Determination of steroid in blood, urine and sweat.
2. Detection of psychotropic drug in plasma.
THE END
Thank You!

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