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3 Acta Chromatographica

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LC-MS/MS method for the quantification of withaferin-A in plant extracts of Withania spp.

Mirjalili M. H., Navarro A., Hernández L., Jáuregui O., Bonfill M.

Acta Chromatographica

2013

Vol. 25, no. 4

745--754

The quantification of the main withanolide, withaferin-A, in plant extracts of Withania somnifera and Withania coagulans using high-performance liquid chromatography (HPLC)-mass spectrometry (ESI-MS/MS) is described. HPLC was used in combination with a triple quadrupole mass spectrometer in multiple reaction monitoring (MRM) mode. Additionally, mass exact determination of withaferin-A was achieved using a hybrid high-resolution mass spectrometer (Q-TOF). A combination of acetonitrile and water as the mobile phase using gradient elution was found to be successful in separating the compound of interest from the other main components. The limit of detection (LOD) in the plants was 6 ng/g and the total analysis time was 10 min.

Validated HPLC method development for simultaneous analysis of withaferin-A and 6-gingerol

Jain V., Thakur A, Soman G, Laddha K. S.

Acta Chromatographica

2010

Vol. 22, no. 1

153--159

A simple, rapid, and specific reversed-phase HPLC method has been developed for simultaneous analysis of withaferin-A and 6-gingerol in a polyherbal formulation containing Withania somnifera and Zingiber officinalis extracts. HPLC analysis was performed on a C 18 column using a 40:60 ( v / v ) mixture of acetonitrile and water as isocratic mobile phase at a flow rate of 1.5 mL min −1 . UV detection was at 227 nm for withaferin-A and 278 nm for 6-gingerol. The method was validated for accuracy, precision, linearity, specificity, and sensitivity in accordance with International Conference on Harmonization guidelines. Validation revealed the method is specific, accurate, precise, reliable and reproducible. Good linear correlation coefficients ( r 2 > 0.9996) were obtained for calibration plots in the ranges tested. Limits of detection were 0.2 and 0.4 μg and limits of quantification were 0.5 and 1.0 μg for withaferin-A and 6-gingerol, respectively. Intra and inter-day RSD of retention times and peak areas were less than 2.1%. Recovery was between 94.5 and 98.8% for withaferin-A and 94.2 and 102.4% for 6-gingerol. The established HPLC method is appropriate and the two markers are well resolved, enabling efficient quantitative analysis of withaferin-A and 6-gingerol. The method was successfully used for quantitative analysis of these two marker constituents in a marketed polyherbal formulation.

RP-HPLC analysis of withanolides in the flowers, leaves, and roots of Withania somnifera

Ali N. W, Abouzid S, Nasib A, Khan S., Qureshi J, Choudhary M. I

Acta Chromatographica

2010

Vol. 22, no. 3

473--480

Ashwaghanda, Withania somnifera , is one of the most widely used herbs in Ayurvedic medicine. Leaves and roots are the traditionally used parts of the plant. An RP-HPLC method using a binary acetonitrile-water gradient containing 0.1% acetic acid has been developed for analysis of withaferin A. The method was validated in accordance with ICH guidelines and used for analysis of the withanolide content of the flowers, leaves, and roots of W. somnifera . The withanolide content was highest in the flowers.