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HPTLC

High-performance thin-layer chromatography (HPTLC) is an advanced form of thin-layer chromatography that enhances resolution and quantification of compounds using automated methods. It has applications in forensic analysis, herbal identification, food industry evaluations, pharmaceutical quality control, and cosmetic product testing. HPTLC offers advantages such as effective separation, reduced analysis time, and the ability to work in conjunction with other analytical techniques.

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106 views6 pages

HPTLC

High-performance thin-layer chromatography (HPTLC) is an advanced form of thin-layer chromatography that enhances resolution and quantification of compounds using automated methods. It has applications in forensic analysis, herbal identification, food industry evaluations, pharmaceutical quality control, and cosmetic product testing. HPTLC offers advantages such as effective separation, reduced analysis time, and the ability to work in conjunction with other analytical techniques.

Uploaded by

brishbhanupriya04
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd

13-11-2022

HPTLC
Chromatography

✓Basic Introduction
✓Principle
✓Procedure
✓Application
✓Advantages

Chromatography
Instrumental Analysis

HPTLC
Introduction:
High-performance thin-layer chromatography (HPTLC) is an enhanced
form of thin-layer chromatography (TLC).
A number of enhancements can be made to the basic method of
thin-layer chromatography to automate the different steps, to
increase the resolution achieved, and to allow more accurate
quantitative measurements.
HPTLC helps in better resolution of compounds with lower limits of
detection and quantifies separated components with the use of an
integrated software platform.

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13-11-2022

HPTLC

HPTLC
Principles:
The principle of separation is adsorption similar as TLC

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HPTLC

Methodology:

HPTLC

Methodology:
Stationary Phase:
Precoated plate (Silica Gel GF254)
Particle size- 7 um,
Thickness- 100 um
Used to separate- Amino acid, alkaloids,
fatty acids, lipids, steroids, terpenoids, etc.
Mobile Phase:
Similar as TLC
Depends on the polarity of analyte

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HPTLC
Methodology:
Sample applicator
Linomate 5 applicator with 100 ul of
micro syringe is used
Nitrogen gas is used to spray, dry or
apply the sample on plate with fixed
pressure (>100 psi)
Software based system for sampling (2-5
ul) with calculated distance, number,
etc.
practical Tutorials: [Link]
[Link]

HPTLC
Methodology:
Visualizer (254 & 366 nm)
[Link]

Scanner
[Link]

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HPTLC
Application:
Forensic Analysis: A challenge in forensic toxicology is the identification of
unknown poisonous substances in intoxication cases. HPTLC offers rapid
identification as well as qualitative and quantitative analysis for toxic
substances.
Herbal Applications: HPTLC fingerprint technology can be used in the
identification of botanical materials that are very complex in nature.
Food Industry: To evaluate nutrients, beverages, vitamins, and pesticides in
fruit, vegetables, and other foodstuffs.
Pharmaceutical Industry: Used in post-production quality control. analysis of
forced degradation studies, stability testing, and to check the presence of
impurities in the drug.

HPTLC
Application:
Cosmetic Industry: HPTLC technique can be employed for the detection of
UV filters used in the formulation of cosmetic products. The identified filter
can be further confirmed using Mass Spectrometer.

Advantages
Most effective separation techniques employed in the discovery,
development, and analysis of new drugs.
More than one analyst can work on the system at a time.
Shorter developing and analysis time.
Fresh stationary and mobile phase is used for each analysis so no
interference or contamination comes in the analytical process.
It can be used alone or in combination with other techniques like MS, FTIR,
Densitometry.

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