Role of HPTLC in Characterization of Herbal Medicines

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Role of HPTLC in Characterization of Herbal Medicines

by | Mar 23, 2023

Modern Analytical Tools used for Characterization of Herbal Medicines

Market scenario of medicinal plants

The market for medicinal plants in India stood at Rs. 4.2 billion (US$ 56.6 million) in 2019 and is expected to increase at a CAGR of 38.5% to Rs. 14 billion (US$ 188.6 million) by 2026.

The total world herbal trade is currently assessed at US$ 120 billion. India’s share in the global export of herbs and herbal products is low due to unsophisticated agricultural and quality control procedures, lack of processing, research & development, standardization in products and regulatory framework in the trade of medicinal plants.

To address this problem it is necessary to standardize medicinal plants and formulations prepared from herbal ingredients by using modern analytical techniques e.g. Chromatography and spectroscopic techniques.

What are the challenges in characterizing herbal medicines?

Herbal medications are difficult to characterize because they are complex mixtures of numerous bioactive compounds that vary greatly in composition, quantity, and biological activity. Plant species, geographical origin, cultivation methods, harvest timing, storage conditions, and processing procedures all contribute to this heterogeneity.

The introduction of substitutes or the unintentional/intentional addition of confounded materials complicates the characterization of herbal medicines.

Mass spectrometry (MS), Nuclear magnetic resonance spectroscopy (NMR), gas chromatography (GC), Fourier transform infrared (FTIR) spectroscopy, Ultra-performance liquid chromatography (UPLC), High-performance liquid chromatography (HPLC), High-Performance thin layer chromatography (HPTLC), and such chromatographic and spectroscopic techniques are used for the characterization of herbal medicines. Of these techniques, HPTLC would be the ideal alternative to standardize many characteristics of herbal medicines since it is an easy, versatile, sensitive, cost-effective, and non-destructive technology that enables high-throughput study of herbal medicines. Its capacity to separate and identify a broad spectrum of compounds, even those present in low concentrations, makes it a powerful tool for herbal medicine characterization.

Role of HPTLC in Characterization of Herbal Medicines

High Performance Thin Layer Chromatography (HPTLC) is one of the best chromatographic techniques for herbal analysis.

  • It is the only chromatographic technique in which we can record the results in the form of images/pictures.
  • HPTLC is a type of planar chromatography technique and it is a GLP/USP/EP compliant chromatographic technique.
  • HPTLC is a simple, flexible, cost-effective and fast technique. More than 100 samples can be analyzed in a day and report can be generated in approximately 60-90 minutes.
  • HPTLC has applications in numerous fields e.g. herbal, food, pharma, cosmetic, forensic etc. However, different categories of herbal samples e.g. AYUSH samples, Nutraceuticals, dietary supplements can also be analyzed.
  • HPTLC has tremendous applications in AYUSH labs, herbal research labs and manufacturing units, academic institutes etc.

What type of characterization studies can be done by HPTLC?

1. Fingerprint studies are conducted for herbal raw materials or finished product to establish the identity of the said material.

2. The correlation between raw material and finished product can be established by an identification test by HPTLC.

3. Stability studies can also be performed by HPTLC using two approaches: fingerprint studies or quantification of phytomarkers present in the formulation.

4. Quantification of respective phytomarkers and method validation studies can also be performed in herbal raw material and finished product for quality testing or optimization procedures of related herbal samples.

5. HPTLC can also be coupled with a mass analyzer for the identification of separated analytes. USP Chapter 2251 states the use of the TLC-MS interface for the identification of synthetic API in herbal formulations.

6. HPTLC can also be coupled with bioactivity studies, e.g., antioxidant, antidiabetic, antimicrobial, etc. These types of studies help with fast screening for a large number of samples in less time and give information about the therapeutic activity of the sample.

Let us see one example in which you can understand the simultaneous identification and quantification of four markers in Withania somnifera aerial and root parts by HPTLC:

Ashwagandha or Withania somnifera also known as ‘Indian ginseng’ is an herb used to calm the brain, reduce swelling, lower blood pressure, treat insomnia, boost nutrition and alter the immune system.

We, at Anchrom have developed a simultaneous method for identification and quantification of Withanoside – IV, Withanoside – V, Withaferin A and Kaempferol-3-7-di-O-glucoside from Withania somnifera aerial parts and root part.

A combination standard mixture was prepared and along with it the samples of aerial parts and root part were prepared and applied on the plate. After development, four respective standards were detected after derivatization with anisaldehyde sulphuric acid in white light.

Following table shows the data for quantitative analysis performed for the samples (aerial parts & root part) provided by the customer.

SampleKaempferol-3-7-di-O-glucosideWithanoside – IVWithanoside – VWithaferin A
Aerial Parts extract 0.21%0.43%
Root part extract0.146%0.153%0.08%

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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