seminar presentation on HPLC principle and instrumentation

1. PRINCIPLE AND INSTRUMENTATION OF HIGH
PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
 Daniyan Temitayo Ezekiel 180403008
 Jegede Justinah Bukola 180402021
 Aladejayan Emmanuel Adedoyin 180402028
 Alaba Iyanuolwa Ayodeji 180402027
 Enegobru Faith Odinrinvherere 180402032
1
ADEKUNLE AJASIN UNVERSITY, AKUNGBA ONDO STATE.
DEPARTMENT OF CHEMISTRY
A SEMINAR PRESENTATION ON:
Guided by:
Prof. Isaac Ayodele Ololade

2. TABLE OF CONTENTS:
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BRIEF HISTORY OF CHROMATOGRAPHY
INTRODUCTION TO THE HPLC
THE PRINCIPLE OF HPLC
INSTRUMENTATION OF HPLC
SAMPLE PREPARATION IN HPLC
APPLICATIONS AND LIMITATIONS OF HPLC

3. BRIEF HISTORY OF CHROMATOGRAPHY 3

4. 4
INTRODUCTION TO HPLC

5. • HPLC - High-performance liquid
chromatography
• is an automated system of
column chromatography which
involves the flow of liquid
(mobile phase) at high pressure
through a column (stationary
phase) in order to separate,
identify, and quantify the active
compounds(analytes).
What
is
HPLC ?
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DIAGRAM OF THE HPLC SYSTEM

7. Identification
of Analyte in a
mixture.
Quantification
of Analyte in a
mixture.
Purification
of Analyte in
a mixture.
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MAIN
PURPOSES
OF USING
HPLC

8. PUMPING
OF MOBILE
PHASE
INJECTION
OF SAMPLE
SEPARATION
OF SAMPLE
CONSTITUENTS
DETECTION
DISPLAY
OF RESULT
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THE PRINCIPLE
OF HPLC

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INSTRUMENTATION OF THE HPLC

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SAMPLE PREPARATION IN HPLC
Sample preparation is the transformation of sample into a form that is suitable for
analysis. It is an essential part of chromatographic analysis.
Steps Involved in Sample Preparation:
 SAMPLING
 FILTRATION
 EXTRACTION
 CONCENTRATION OR DILUTION
The purpose of sample preparation is to:
(a) Improve the quality or suitability of the sample to be analysed.
(b) Obtain a reasonable analytical result.
Types of sample:
Sample matrices can be classified as Organic
and Inorganic. It can further be divided into;
 Solids
 Semisolids (gels, cream and suspensions)
 Liquids
 Gases.

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APPLICATIONS OF THE HPLC
HPLC has helped to develop analytical solutions in a variety of fields, including pharmaceutics, food, life
science, the environment, forensics, etc.
 HPLC is used in drug analysis: identifies APIs,
controls stability and shelf life, quality assurance
etc.
 HPLC is used in Food analysis for specific product
research and quality control, to identify dietary
additives and pollutants, etc.
 HPLC is used in finding anabolic steroids in urine,
sweat, hair, and serum.
 It is also used in blood test analysis to find cocaine
and its metabolites.
 HPLC is used in clinical research and analysis,
determination of poisoning origin, and
medication metabolism, etc.
PHARMACOLOGICAL APPLICATIONS: CLINICAL APPLICATIONS:
FORENSICS APPLICATIONS: FOOD APPLICATION:

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1. SAMPLE SIZE: Challenge of scarce or expensive samples.
2. COMPLEXITY: Challenges in operating the Instrument because the HPLC is a complex technique.
3. DETECTION LIMIT: HPLC has a lower detection limit compared to other analytical methods like GC or UV.
4. COLUMN LIFETIME: HPLC columns must occasionally be replaced because they are consumable items.
5. COST PRICE: HPLC is a pricey technology because it needs specialized equipment and columns.
LIMITATIONS OF THE HPLC

13. THE END
THANK YOU SO MUCH FOR LISTENING…
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