LimsPulse
Chromatography

High-Performance Thin-Layer Chromatography (HPTLC)

An instrumental, standardised form of TLC that uses fine-particle plates, automated band application, controlled development and densitometric scanning. Widely used for herbal identification fingerprints and semi-quantitative or quantitative assays.

  • Pharmaceutical
  • Herbal and nutraceutical
  • Food & cosmetics
  • Forensic
  • Academic research
Photo of a High-Performance Thin-Layer Chromatography (HPTLC)
Photo: Oak Ridge National Laboratory, CC BY 2.0 (cropped), via Wikimedia Commons
LEARN & PRACTICE

Test your HPTLC knowledge

Multiple-choice questions

Pick an answer to see whether you are right and why.

  1. Q1. Compared with classical TLC plates, HPTLC plates have:

    basic

  2. Q2. A zone moves 28 mm and the solvent front moves 70 mm from the application line. The Rf is:

    basic

  3. Q3. Why are samples applied as narrow bands rather than spots in HPTLC?

    intermediate

  4. Q4. Which factor most strongly affects Rf reproducibility on silica gel?

    intermediate

  5. Q5. What is the main pharmacopoeial use of HPTLC for herbal drugs?

    basic

  6. Q6. Which instrument measures the absorbance or fluorescence of each zone along a track?

    intermediate

Viva questions

Think of your answer first, then open it to compare.

  1. 1. How is HPTLC different from TLC, and why does it give more reproducible results?

    HPTLC uses finer, uniform plates and instruments for every step: automatic band application, controlled development (saturation and humidity), standardised derivatisation and calibrated documentation and scanning. Controlling these variables makes results comparable between plates and labs.

  2. 2. Why is a system suitability reference applied on every HPTLC plate?

    Each plate is its own run. The reference must show the specified zones and separation, proving that plate and development worked correctly before samples on it are evaluated.

  3. 3. Why is a plate documented before and after derivatisation?

    Some compounds are visible only under UV 254 or 366 nm before derivatisation, and others only after the reagent reacts. Documenting both captures the full fingerprint, and colours after derivatisation can fade, so images are taken promptly.

In LimsPulse · Calibration Management

Keep every HPTLC in control: usage logs, calibration and maintenance schedules, and automatic blocking of overdue instruments.

See how