How is HPTLC different from TLC?

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How is HPTLC different from TLC?

by | Dec 28, 2022

Thin Layer Chromatography is a separation technique which is used to separate mixtures of organic compounds. This technique is in practice since 1960 and is been used for the separation of a lot of different compounds.

Thin Layer Chromatography technique and its Limitations:

Thin Layer chromatography is a manual technique. Nowadays some automation is introduced but still, complete automation is not there. The step-like sample application is done with capillary spotting, spotting causes a lot of diffusion and results in poor separation.

Plate development is done in a beaker or round bottom chamber and after development results are seen in the UV cabinet. If required derivatization is done by manual spraying which causes a lot of dripping of the reagent on the TLC plate and sometimes affects the resolution of the separated analyte.

Quantification is not possible because of poor separation and uncontrolled sample application. Besides this, there are a lot of environmental factors like temperature, humidity etc. which also contribute to poor separation. It does not meet modern regulatory requirements.

Traditional TLC is not suitable for QC because of several reasons such as noncompliance with GLP, non-validated and qualitative or semi-quantitative methods, separation in non-equilibrated conditions etc. Still, the TLC technique is popular because of its simplicity and fast results.

HPTLC technique, features and advantages:

HPTLC (High-Performance Thin Layer Chromatography) is a modern version of TLC wherein the principle remains the same but the practice is fully automated and GLP/USP/EP and 21 CFR rule 11 compliant. HPTLC practice is standardized by United States Pharmacopoeia and the focus of HPTLC is on reproducibility and high separation power.

Advantages of HPTLC over TLC:

  • HPTLC can be used for both qualitative and quantitative analysis of different samples e.g. Herbal, food, pharma, forensic, cosmetic, speciality chemicals etc.
  • HPTLC is a high throughput technique, more than 100 samples can be run per day with minimal analysis cost.
  • HPTLC enables multiple detections without repeating chromatography which means one can get many data points after doing a single analysis which gives thorough information about the sample applied.

Response of the analyte can be checked in UV, fluorescence and white light with a suitable TLC photo documentation unit. Chromatograms can be obtained with CAMAG scanner 4 for quantitative analysis and spectral evaluation. If required derivatization can be done with CAMAG derivatizer.

  • HPTLC can also be coupled with a mass analyser for the identification of separated analytes by means of a TLC-MS interface.
  • The results obtained are highly reproducible, reliable analytical results and come with traceable data and images.
  • Highly standardized parameters are used for HPTLC analysis. HPTLC methods can be developed and validated as per suitable guidelines.
  • HPTLC can also be coupled with biological detection methods e.g. antioxidant activity, antidiabetic activity, antimicrobial activity etc.
  •  HPTLC has applications in the herbal, food, pharma, forensic and cosmetic field.

All the above features make HPTLC a unique, more advantageous and reliable analytical technique.

Anchrom Enterprises Pvt. Ltd is one of the leaders in HPTLC analysis. Please contact us at for HPTLC analysis of plant extracts, drugs, ingredients in cosmetics, and forensic science.


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