8/13/22, 9:40 AM What is the Difference between HPLC and HPTLC?

What is the Difference between HPLC and HPTLC?

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HPLC and HPTLC are two types of automated separation techniques with similar
principles of separation. Furthermore, HPLC consists of a pump-driven flow system
through the stationary phase filled in a column while HPTLC is a type of planar
chromatography in which the solvent moves through a stationary phase fixed on a
plate. Moreover, HPLC consists of a closed system, while HPTLC is an open system.
Both HPLC and HPTLC are the separation techniques of chromatography operating on
the same principles of separation.

What is HPLC? 
HPLC is known as high-pressure liquid chromatography and is also known as High-
performance liquid chromatography, it is an analytical technique, that is used to
separate, determine, and identify each component in a complex mixture. 
It depends on a pressurized mobile phase provided by the pumps in which the sample
mixture is included and passes through the HPLC column. Every analyte in the
complex mixture interacts with the absorbing material in a different way, which has
dissimilar flow rates for the different analytes and leads to the separation of the analyte
or components.

What is HPTLC?
High-performance thin-layer chromatography is the advanced form of TLC and
contains the use of chromatographic layers of cutting-edge separation efficiency for all
stages of the process and the use of state-of-the-art instrumentation, reproducible
chromatogram, accurate sample application, and software-based evaluation. 
HPTLC is a complete concept in which a broad, standardized method depends on
scientific facts, and the quantitative and qualitative analysis uses validated methods.

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8/13/22, 9:40 AM What is the Difference between HPLC and HPTLC?

The main difference between HPLC and HPTLC is that HPLC permits quantitative
separation of analytes in a sample of a complex mixture, whereas HPTLC does not allow it.

What is the principle of HPTLC?

The principle of HPTLC has a similar approach and employs the same physical
principles of TLC (adsorption chromatography) i.e. the principle of separation is
adsorption. The mobile phase solvent flows through because of capillary action. The
components move according to their affinities towards the adsorbent.
HPTLC (high-performance thin-layer chromatography) is a sophisticated form of TLC,
which provides superior separation efficiency. The HPTLC concept includes validated
methods for qualitative and quantitative analysis and fulfills all quality requirements
for use in fully regulated environments.
The CAMAG TLC Scanner 4 (densitometer) is the most advanced workstation for
densitometric evaluation of TLC/HPTLC chromatograms and other planar objects. The
chromatogram is evaluated under white or UV light.

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8/13/22, 9:40 AM What is the Difference between HPLC and HPTLC?

What is the principle of HPLC?

HPLC is a highly improved form of column chromatography. A pump forces a solvent
through a column under high pressures of up to 400 atmospheres. The column packing
material or adsorbent or stationary phase is typically a granular material made of solid
particles such as silica or polymers.

Why is HPLC used?

HPLC is used to split a mixture of compounds in the fields of analytical chemistry,
biochemistry, Pharmaceutical, and industrial. The main purposes of using HPLC are
for identifying, quantifying, and purifying the individual components of the mixture.

Applications of HPLC
Water purification. 
Detection of impurities in pharmaceutical industries. 
Pre-concentration of trace components. 
Ligand-exchange chromatography.

HPTLC vs. HPLC

High-performance thin-layer chromatography (HPTLC) is still increasingly finding its way
into pharmaceutical analysis in some parts of the world. With the advancements in the
stationary phases and the introduction of densitometers as detection equipment, the
technique achieves for given applications a precision and trueness comparable to high-
performance liquid chromatography (HPLC). 

Basic differences between HPLC and HPTLC are given in the following table.

Parameters HPLC HPTLC

Type Reverse
Phase Straight
Phase
Chromatography Chromatography

Stationary
phase Liquid Solid

Conditioning
phase None Gas

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8/13/22, 9:40 AM What is the Difference between HPLC and HPTLC?

Separation
by Partition Adsorption

Results By
machine By
machine + eyes

Analysis On
‐ line Off
‐ line

Resolution Very
high Moderate
to high

Chromatography
System Closed Open

Separating
medium Tubular
column Planar
layer (plate)

Strongly
Retarded Broad
peaks Sharp
Peaks

Fractions
Seen As

Analysis
in parallel No. Yes.

Only
1 at a time Upto
100 samples.

High
temp. / pressure High
pressure None

Time
per sample 2‐
60 min 1-30
min

Data
obtained from Limited
to very high High
to very high
chromatography

Post chromatography Limited possibilities. Simple. Possible for every

derivatisation Cumbersome. sample. Gives additional
information

Fraction collection / micro Requires prep. scale Simple. No special

preparative chromatograph & fraction requirements
chromatography collector

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8/13/22, 9:40 AM What is the Difference between HPLC and HPTLC?

Sensitivity High to ultra high Moderate to ultrahigh

Fluorescence data Possible, optional Possible, built-in

Detectors UV, Fluor, electrochem UV ‐ Vis,

bioluminescence, MS
Light

scatter, MS

Chromatogram image No Yes. At 254 & 366 nm &

documentation visible

Sample clean-up Through Column reusable Not so imp.

Layer disposable

Chromatographic Yes, but limited Yes. Comprehensive

fingerprint

Cost per analysis Very high Low

Eqpt. maintenance Very high Low

Analyst’s skills required High to very high Low (TLC) to high (HPTLC)

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