Separation, identification, and quantification of selected withanolides in plant extracts of Withania somnifera by HPLC‐UV(DAD) – Positive ion electrospray ionisation–mass spectrometry
- 7 May 2004
- journal article
- research article
- Published by Wiley in Journal of Separation Science
- Vol. 27 (7-8), 541-546
- https://doi.org/10.1002/jssc.200301690
Abstract
This paper describes a method for separation, identification, and quantification of selected withanolides in Withania somnifera plant extracts by HPLC‐UV(DAD)–Mass Spectrometry (HPLC‐MS). Withaferin‐A (WS‐3), 12‐deoxywithastramonolide (WS‐12DS), Withanolide A (WS‐1), and Withanone (WS‐2) were used as external standards. The compounds were isolated from Withania somnifera by repeated column chromatography of the root extract and their identity was established by 1H‐ and 13C‐NMR and mass spectral data. The compounds were chromatographed on a Merck (250×4.6 mm ID, 5 μm) column and analyzed by Electrospray Ionization on a mass spectrometer in Selected Ion Mode (SIM). For quantification, [M + Na]+ ions were monitored. Linear calibration curves were obtained in the concentration range of 1.50 μg/mL to 6.5 μg/mL. The method was applied successfully to the detection and quantification of the said withanolides in a number of samples.Keywords
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