HPTLC & AYURVEDA

Stationary Phase: 

This is the fixed component of the chromatographic system. It can be a solid, a gel, or a liquid coated on a solid support. The substances in the mixture will interact with this phase through various mechanisms like adsorption, partitioning, ion exchange, or size exclusion.

Slica gel - F264 in varied dimensions 10*10 cm, 20*10 cm, and 20*20 cm. Based on number of Samples for the study Size of the Plate is taken, 7, 15 and 15/30 samples respectively.

Mobile Phase: 

This is the fluid (liquid) that moves through or over the stationary phase, carrying the components of the mixture with it inside a closed chamber.

Fluid/solvent composition varies depending on the phytoconstituent, plant, formulation, or study material based on its absorption, solubility, polarity etc natures of the molecules/compounds.

Common solvents used in HPTLC are Methanol, Ethanol, Ethyl Acetate, Choloroform, Toulene, Formic Acid, Acetic Acid etc.

Step 1: Sample Preparation

Study sample is taken in a very fine powder form and filled belown 1/4th volume of centrifuge tube., with a suitable solvent system ( Commonly Water, Methanol, Hydroalcoholic solvents are used )

Using Mechanical energy equipments, like Vortex, sonicator, Orbital Spinner etc Actives is made soluble in the solvent and centrifuges at 3000 rpm for 10 minutes.

Clear Sample is trasferred to 2ml vials with the help of pippetes and placed on Sample applicator equipment.

Sample Applicator can be programmed to Spray precise ammout of sample between 1 um to 10 um onto TLC Plate.  

Step 2: Sample Application

Once the sample is prepared, it will be mounted on racks and applied using automatic applicator onto TLC Plate with precised measurements.

Step 3: HPTLC Development

TLC plate will be placed inside the saturated twin-thru chamber with suitable mobile phase till mobile phase carries and seperates the mixture componants of a sample. Once done plate is taken out and dried.

Step 4: Visualization 

Using TLC Visualizer, plate is visualized at various light waves (366nm, 254nm, white, white-rt)

If bands are not visible, further dervitization is done.

Step5: Quantification/ Profile Generation

Once the above steps are completes, and proper seperation is acheived, plate is placed in densitometric scanner to obtain peaks.

•  Clinical application - for Metabolism studies 
•  Drug Screening 
•  Pharmaceutical - Quality control 
•  Purity Analysis 
•  Forensics - Poisoning Investigations 
•  Food Analysis - QC/ Additives/ Pesticides/ Stability test 
•  Bio-availability Studies 
•  Fingerprint profiling - Identification/ Plant Research  

•  Clinical application - for Metabolism studies 
•  Drug Screening 
•  Pharmaceutical - Quality control 
•  Purity Analysis 
•  Forensics - Poisoning Investigations 
•  Food Analysis - QC/ Additives/ Pesticides/ Stability test 
•  Bio-availability Studies 
•  Fingerprint profiling - Identification/ Plant Research