HPTLC Quantification of Withaniasomnifera (L.) Dunal


1Raman Preet*, 2Lata

Bulletin of Pure and Applied Sciences.

Botany, Vol.41 B, No.2.
July-December  2022: P.117-125

Original Research Article

DOI: 10.5958/2320-3196.2022.00014.3



HPTLC Quantification of Withaniasomnifera (L.) Dunal

 1Raman Preet*, 2Lata

Author’s Affiliation

1Assistant Professor, Department of Botany, DAV College, Abohar, Punjab 152116, India

2Assistant Professor, Department of Botany, Eternal University, Baru Sahib, Himachal Pradesh, India


Raman Preet

Assistant Professor, Department of Botany,

DAV College, Abohar, Punjab 152116, India



How to cite this article: Preet R., Lata (2022). HPTLC Quantification of Withaniasomnifera (L.) Dunal. Bulletin of Pure and Applied Sciences-Botany, 41B(2), 117-125.

Received on 15.08.2022

Revised on 19.11.2022

Accepted on 30.11.2022

Published on 15.12.2022

At present study a simple, selective, easy, robust, sensitive and precise method is developed to investigate the outcomes of HPTLC for the simultaneous determination of 3 key withanolides, namely withaferin A, withanolide A and withanolide B  in Ashwagandha; Withaniasomnifera plant part samples. The method is analyzed simultaneously on Camag aluminum per-coated silica gel 60-F254 HPTLC plates, using toluene: ethylacetate:formic acid:ethanol (6:3:0.1:0.6 v/v/v/v) as mobile phase. The Quantitative evaluation of withanolides was performed by densitometry in the reflection-absorption mode at 530 nm. The method was validated for recovery, precision, accuracy, robustness, limit of detection, limit of quantitation, and specificity conferring to International Conference on Harmonization guidelines. Specificity of quantification was confirmed using retention factor (Rf) values, UV Visual spectral correlation, and electrospray ionization mass spectra of marker compounds in sample tracks. This combination of techniques has been applied, for the first time, for the estimation of withaferin-A, withanolide A and withanolide B in fruit, leaf, stem and root parts samples of Withaniasomnifera. Present method may be beneficial in a quality control setting as it would allow rapid qualitative analysis of herbal material while sustaining accurate quantification of extract composition and can be used for routine analysis as well as for quality control of raw materials and herbal formulations. Keywords: Thin Layer Chromatography; Withaniasomnifera; Withaferin A, Withanolide A; Withanolide B