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
Follow your site SOP
This is a typical operating sequence for learning. Your instrument model and approved SOP take precedence.
STAGE 1

Before you start

  1. 1Check calibration status of the applicator, scanner and balance.
  2. 2Use plates from a qualified supplier; pre-wash with methanol and activate if the method requires.
  3. 3Prepare the mobile phase fresh and in the exact proportions stated.
  4. 4Record laboratory temperature and relative humidity.
  • Handle plates by the edges with gloves; fingerprints and dust show up as zones.
STAGE 2

Application and development

  1. 1Set the application pattern: band length, distance from the lower edge (e.g. 8 mm) and from the sides.
  2. 2Apply samples, reference standards and the system suitability mixture on the same plate.
  3. 3Condition the plate to the specified humidity if required, then saturate the chamber (often 20 minutes with filter paper).
  4. 4Develop to the specified distance, then dry the plate in a stream of cold air.
  • Do not let the solvent level reach the applied bands; they would dissolve into the solvent.
STAGE 3

Detection and evaluation

  1. 1Document the plate under UV 254 nm and 366 nm before derivatisation.
  2. 2Derivatise as per the method, heat if required, and document again under white light and/or UV 366 nm.
  3. 3Check the system suitability zones are present and separated as specified.
  4. 4Compare sample fingerprints with the reference, or scan and quantify against a standard calibration.
  • Document derivatised plates promptly; colours can fade or change with time.
STAGE 4

Shutdown

  1. 1Clean the applicator syringe and rinse the chamber.
  2. 2Archive the plate images and raw data; store or dispose of the plate as per SOP.
  3. 3Complete the instrument logs.
In LimsPulse · Calibration Management

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

See how