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3DJH RI
c) Procedure:
20
µ
l of each individual standard solution, mixed standard solutions, sample
solutions were injected into two different HPLC’s [(i) LC2010A equipped with UV detector
and (ii) LC10ADVP equipped with PDA detector)] on different date and time and the
chromatograms were recorded. Relative retention time of different withanolides were
calculated with respect to Withanolide A
.
.
4.1.5) Observation:
Retention time of Withanoside IV is about 15.5 minutes, Withanoside V is
about 19.7 minutes, Withaferin A is about 20.3 minutes, 12-Deoxy withastramonolide is
about 21.4 minutes, Withanolide A is about 22.2 minutes and Withanolide B is about 25.5
minutes (Refer chromatogram-1). In the sample chromatogram all the individual
withanolides are well resolved. (Refer chromatogram-2).
Chromatogram – 1: Standard mix chromatogram
Chromatogram – 2: Sample chromatogram