Separation, identification, and quantification of selected withanolides in plant extracts of Withania somnifera by HPLC-UV(DAD) – Positive ion electrospray ionisation–mass spectrometry
✍ Scribed by Ravi K. Khajuria; Krishan A. Suri; Rajinder K. Gupta; Naresh K. Satti; Musarat Amina; Om P. Suri; Ghulam N. Qazi
- Publisher
- John Wiley and Sons
- Year
- 2004
- Tongue
- English
- Weight
- 593 KB
- Volume
- 27
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
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 ^1^H‐ and ^13^C‐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.