Abstract
Withania somnifera (L.) Dun. (Solanaceae), known as Indian ginseng, is one of the most popular medicinal plants in India. Considering the importance and common use of this plant, it is necessary to investigate its holistic metabolite profile. However, with existing analytical methods which are based on TLC and HPLC-UV (or MS), it is difficult to obtain information of the whole range of compounds appropriately. In this study, the metabolic characterization of Withania somnifera leaves, stems, and roots collected in six different regions in India was performed using 1 H NMR spectroscopy followed by principal component analysis (PCA) and hierarchical clustering analysis (HCA). Of the parts of Withania somnifera analyzed in this study, the leaf was found to have the widest range of metabolites, including amino acids, flavonoids, lipids, organic acids, phenylpropanoids, and sugars, as well as the main secondary metabolites of the plant, withanolides. The 1 H NMR spectra revealed the presence of two groups of withanolides: 4-OH and 5,6-epoxy withanolides (withaferin A-like steroids) and 5-OH and 6,7-epoxy withanolides (withanolides A-like steroids). The ratio of these two withanolides was found to be a key discriminating feature of Withania somnifera leaf samples from different origins.
Key words
Withania somnifera
- Solanaceae - metabolic characterization - NMR - withanolides
References
1
Kulkarni S K, Dhar A.
Withania somnifera : an Indian Ginseng.
Prog Neuropsycho pharmacol Biol Psychiatry.
2008;
32
1093-1105
2
Singh B, Saxena A K, Chandan B K, Gupta D K, Bhutani K K, Anand K K.
Adaptogenic activity of a novel withanolide-free aqueous fraction from the roots of Withania somnifera Dun.
Phytother Res.
2001;
15
311-318
3
Kulkarni S K, George B, Mathur R.
Neuroprotection by Withania somnifera root extract against lithium-pilocarpine-induced seizures.
Indian Drugs.
1998;
35
208-215
4
Bhattacharya S K, Goel R K, Kaur R, Ghoshal S.
Anti-stress activity of sitoindosides VII and VIII new acyl steryl glycosides from W. somnifera .
Phytother Res.
1987;
1
32-37
5
Mishra L C, Singh B B, Dagenais S.
Scientific basis for the therapeutic use of Withania somnifera (ashwagandha): a review.
Altern Med Rev.
2000;
5
334-346
6
Elsakka M, Grigorescu E, Stanescu U, Stanescu U, Dorneanu V.
New data referring to chemistry of Withania somnifera species.
Rev Med Chir Soc Med Nat Iasi.
1990;
94
385-387
7
Dhar R S, Verma V, Suri K A, Sangwan R S, Satti N K, Kumar A, Tuli R, Qazi G N.
Phytochemical and genetic analysis in selected chemotypes of Withania somnifera .
Phytochemistry.
2006;
67
2269-2276
8
Matsuda H, Murakami T, Kishi A, Yoshikawa M.
Structures of withanosides I, II, III, IV, V, VI, and VII, new withanolide glycosides, from the roots of Indian Withania somnifera DUNAL and inhibitory activity for tachyphylaxis to clonidine in isolated guinea-pig ileum.
Bioorg Med Chem.
2001;
9
1499-1507
9
Sangwan R S, Chaurasia N D, Misra L N, Lal P, Uniyal G C, Sharma R, Sangwan N S, Suri K A, Qazi G N, Tuli R.
Phytochemical variability in commercial herbal products and preparations of Withania somnifera (Ashwagandha).
Curr Sci.
2004;
86
461-465
10
Dhar R S, Verma V, Suri K A, Sangwan R S, Satti N K, Kumar A, Tuli R, Qazi G N.
Phytochemical and genetic analysis in selected chemotypes of Withania somnifera .
Phytochemistry.
2006;
67
2269-2276
11
Chaurasiya N D, Uniyal G C, Lal P, Misra L N, Sangwan N S, Tuli R, Sangwan R S.
Analysis of withanolides in root and leaf of Withania somnifera by HPLC with photodiode array and evaporative light scattering detection.
Phytochem Anal.
2008;
19
148-154
12
Yang S Y, Kim H K, Lefeber A W M, Erkelens C, Angelova N, Choi Y H, Verpoorte R.
Application of two-dimensional nuclear magnetic resonance spectroscopy to quality control of ginseng commercial products.
Planta Med.
2006;
72
364-369
13
Kim H K, Choi Y H, Erkelens C, Lefeber A W M, Verpoorte R.
Metabolic fingerprinting of Ephedra species using 1 H-NMR spectroscopy and principal component analysis.
Chem Pharm Bull.
2005;
53
105-109
14
Choi Y H, Kim H K, Hazekamp A, Erkelens C, Lefeber A W M, Verpoorte R.
Metabolomic differentiation of Cannabis sativa cultivars using 1 H NMR spectroscopy and principal component analysis.
J Nat Prod.
2004;
67
953-957
15
Kim H K, Khan S, Wilson E G, Kricun S D P, Meissner A, Göraler S, Deelder A M, Choi Y H, Verpoorte R.
Metabolic classification of South American Ilex species by NMR-based metabolomics.
Phytochemistry.
2010;
71
773-784
16
Chatterjee S, Srivastava S, Khalid A, Singh N, Sangwan R S, Sidhu O P, Roy R, Khetrapal C L, Tuli R.
Comprehensive metabolic fingerprinting of Withania somnifera leaf and root extracts.
Phytochemistry.
2010;
71
1085-1094
17
Kim H K, Choi Y H, Verpoorte R.
NMR-based metabolomics of plants.
Nat Protoc.
2010;
3
536-549
18
Nittala S S, Van de Velde V, Frolow F, Lavie D.
Chlorinated withanolides from Withania somnifera and Acnistus breviflorus .
Phytochemistry.
1981;
20
2547-2552
19
Lavie D, Glotter E, Shvo Y.
Constituents of Withania somnifera Dun. III. The side chain of withaferin A.
J Org Chem.
1965;
30
1774-1778
20
Subramanian S S, Sethi P D, Glotter E, Kirson I, Lavie D.
5,20α (R)-dihydroxy-6α ,7α -epoxy-1-oxo-(5α ) witha-2,24-dienolide, a new steroidal lactone from Withania coagulans .
Phytochemistry.
1971;
10
685-688
21 Eriksson L, Johansson E, Kettaneh-Wold N, Wold S. Multi- and megavariate data analysis. Umeå: Umetrics Academy; 2001
22
Bown A W, MacGregor K B, Shelp B J.
Gamma-aminobutyrate: defense against invertebrate pests?.
Trends Plant Sci.
2006;
11
424-427
23
Lubbe A, Pomahačová B, Choi Y H, Verpoorte R.
Analysis of metabolic variation and galanthamine content in Narcissus bulbs by 1 H NMR.
Phytochem Anal.
2010;
21
66-72
24
Ganzera M, Choudhary M I, Khan I A.
Quantitative HPLC analysis of withanolides in Withania somnifera .
Fitoterpia.
2003;
74
68-76
25
Abraham A, Kirson I, Glotter E, Lavie D.
A chemotaxonomic study of Withania somnifera (L.) Dun.
Phytochemistry.
1968;
7
957-962
Dr. Young Hae Choi
Division of Pharmacognosy, Section Metabolomics Institute of Biology Leiden University
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