In:
Pharmacology, S. Karger AG, Vol. 89, No. 5-6 ( 2012), p. 260-269
Abstract:
〈 i 〉 Aims: 〈 /i 〉 We wanted to investigate the possible antithrombotic effects and elucidate the chemical identity of the active principles involved in inhibitory effects against adenosine diphosphate (ADP)-induced aggregation of human platelets by wild garlic, 〈 i 〉 Allium ursinum 〈 /i 〉 L. 〈 i 〉 Methods: 〈 /i 〉 For this purpose, a bioassay-guided isolation procedure was used followed by spectrometric identification of pure active compounds. For the bioassay, blood was taken from healthy human volunteers and platelet-rich plasma was prepared for turbidimetric platelet aggregation tests. Platelet-rich plasma, stimulated with 20 µmol/l of ADP, was treated with extracts of different polarities, fractions and isolated single compounds from 〈 i 〉 A. ursinum 〈 /i 〉 . The extracts were investigated by thin-layer chromatography (TLC), HPLC, mass spectroscopy, electrospray ionization mass spectrometry (ESI-MS) and 1/2-dimensional 〈 sup 〉 1 〈 /sup 〉 H/ 〈 sup 〉 13 〈 /sup 〉 C-nuclear magnetic resonance (NMR) spectroscopic techniques. 〈 i 〉 Results: 〈 /i 〉 Fresh 〈 i 〉 A. ursinum 〈 /i 〉 leaves were extracted with ethanol, which was the potent form that effectively inhibited ADP-induced aggregation of human platelets. This ethanolic extract was subjected to liquid-liquid partition. Whilst the aqueous phase, containing the moiety of cysteine sulphoxide and thiosulphinate derivatives, showed only weak activity on platelet aggregation, the ethyl-acetate and particularly the chloroform partitions showed the highest aggregation-inhibiting potency. Thus, in our bioassay, the effects of alliins/allicins could be neglected. The chloroform phase, possessing the strongest activity, was separated into 28 fractions by gradient-elution open column chromatography on silica gel. The most active fractions 11–17 were separated again, yielding 10 subfractions. This afforded 1,2-di-O-α-linolenoyl-3-O-β-D-galactopyranosyl-sn-glycerol and β-sitosterol-3-O-β-D-glucopyranoside, the structures of which were determined by ESI-MS and 1/2-dimensional 〈 sup 〉 1 〈 /sup 〉 H/ 〈 sup 〉 13 〈 /sup 〉 C NMR spectroscopic techniques. Furthermore, the minute amounts of volatile oil of 〈 i 〉 A. ursinum 〈 /i 〉 leaves obtained by steam distillation according to Ph. Eur. could be evaluated as a third aggregation-inhibiting principle. 〈 i 〉 Conclusion: 〈 /i 〉 In our study, for the first time, 2 active, non-sulphur-containing constituents of wild garlic, namely a galactolipid and a phytosterol, could be identified exhibiting inhibitory action on ADP-induced aggregation in human blood platelets. As a major constituent, the galactolipid, 1,2-di-O-α-linolenoyl-3-O-β-D-galactopyranosyl-sn-glycerol, not yet found in 〈 i 〉 Allium 〈 /i 〉 sp., appears as a new, highly useful marker substance for 〈 i 〉 A. ursinum 〈 /i 〉 drugs, or their pharmaceutical or food preparations, as shown by our orientating TLC analyses.
Type of Medium:
Online Resource
ISSN:
0031-7012
,
1423-0313
Language:
English
Publisher:
S. Karger AG
Publication Date:
2012
detail.hit.zdb_id:
1483550-2
SSG:
15,3
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