Analysis of withanolides in root and leaf of Withania somnifera by HPLC with photodiode array and evaporative light scattering detection.
A reversed-phase HPLC method for the simultaneous analysis of nine structurally similar withanolides, namely, 27-hydroxy withanone, 17-hydroxy withaferin A, 17-hydroxy-27-deoxy withaferin A, withaferin A, withanolide D, 27-hydroxy withanolide B, withanolide A, withanone and 27-deoxywithaferin A, has been developed using a linear binary gradient solvent system comprising methanol and water containing 0.1% acetic acid. Both photodiode array and evaporative light scattering detection were used to profile the extract compositions and to quantify the withanolides therein. Homogeneity and purity of each peak was ascertained by comparative evaluation of the on-line UV spectra of the eluted compounds with those of the reference compounds. The method has been validated with respect to various parameters of performance quality including computation regression analysis based on calibration curves, peak resolution factor, asymmetry factor, tailing factor, RSD (%) of retention time and peak area response, limit of quantivation, limit of detection, precision and recovery. The developed method has been applied to the analysis of leaf and root tissues of Withania somnifera for withanolide content.
Chaurasiya
Narayan Das
ND
Central Institute of Medicinal and Aromatic Plants, PO CIMAP, Lucknow-226015, India.
Uniyal
Girish Chandra
GC
Lal
Payare
P
Misra
Laxminarain
L
Sangwan
Neelam Singh
NS
Tuli
Rakesh
R
Sangwan
Rajender Singh
RS
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015394:Molecular Structure; D018515:Plant Leaves / Q000737:chemistry*; D018517:Plant Roots / Q000737:chemistry*; D032341:Withania / Q000737:chemistry*; D054358:Withanolides / Q000032:analysis* / Q000737:chemistry
D054358:Withanolides