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What is the Difference between HPLC and HPTLC?

  • 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.

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.

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.

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.





Reverse Phase Chromatography

Straight Phase Chromatography

Stationary phase



Conditioning phase



Separation by




By machine

By machine + eyes


On line

Off line


Very high

Moderate to high

Chromatography System



Separating medium

Tubular column

Planar layer (plate)

Strongly Retarded

Fractions Seen As

Broad peaks

Sharp Peaks

Analysis in parallel


Only 1 at a time


Upto 100 samples.

High temp. / pressure

High pressure


Time per sample

2 60 min

1-30 min

Data obtained from chromatography

Limited to very high

High to very high

Post chromatography derivatisation

Limited possibilities. Cumbersome.

Simple. Possible for every sample. Gives additional information

Fraction collection / micro preparative chromatography

Requires prep. scale chromatograph & fraction collector

Simple. No special requirements


High to ultra

high Moderate to ultrahigh

Fluorescence data

Possible, optional

Possible, built-in


UV, Fluor, electrochem Light

scatter, MS

UV Vis, bioluminescence, MS

Chromatogram image



Yes. At 254 & 366 nm &


Sample clean-up

Through Column reusable

Not so imp.

Layer disposable

Chromatographic fingerprint

Yes, but limited

Yes. Comprehensive

Cost per analysis

Very high


Eqpt. maintenance

Very high


Analyst’s skills required

High to very high

Low (TLC) to high (HPTLC)

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