Isolation and characterization of free radical scavenging flavonoid glycosides from the flowers of Spartium junceum by activity-guided fractionation

doi:10.1016/S0378-8741(00)00327-5

Journal of Ethnopharmacology

Volume 73, Issue 3, December 2000, Pages 471-478

https://doi.org/10.1016/S0378-8741(00)00327-5 Get rights and content

Abstract

Spartium junceum L. (Fabaceae) flowers are used for the treatment of peptic ulcers in Turkish folk medicine. The possible superoxide dismutase-like activity of the extracts, fractions and constituents obtained through activity-guided fractionation were studied by using in vitro electron spin resonance spectrometry, in order to explain the role of antioxidant principles in the potent antiulcerogenic activity of the extract. Despite the fact that the triterpene, spartitrioside, which was previously reported as the active antiulcerogenic constituent of the flowers was found almost inactive, the flavonoid-rich fractions showed potent antioxidant activity. Five flavonoid glycosides bearing catechol structure in ring B were isolated from the butanol extract and their structures were elucidated using ¹H- and ¹³C-NMR techniques as isoquercitrin (quercetin 3β-glucoside) (1,); luteolin 4’β-glucoside (2); quercetin 3, 4′-diglucoside (3); azaleatin 3β-glucoside (quercetin 5-methylether 3β-glucoside) (4), quercetin 4′β-glucoside (5). Flavonoids (2) and (4) showed the highest in vitro antioxidant activity with 22.59 and 19.08 U/ml, respectively.

Introduction

Spartium junceum L. (Fabaceae) flowers are reported to be used for the treatment of peptic ulcers in Turkish folk medicine (Yeşilada et al., 1993). In a previous study, antiulcerogenic activity of the flowers was reported (Yeşilada et al., 2000) and a triterpene saponin ‘spartitrioside’ was isolated as the active constituent through in vivo bioassay-guided fractionation and its structure was elucidated by spectral techniques (Yeşilada and Takaishi, 1999).

Recently, the role of active oxygen species in the pathogenesis of various gastric mucosal injuries have been indicated and some exogenous antioxidants including superoxide dismutase (SOD), a superoxide scavenger and allopurinol, a xanthine oxidase inhibitor, were reported to show in vivo protective effect against acute gastric mucosal injuries induced by active oxygen species (Perry et al., 1986, Pihan et al., 1987) as well as treating patients with peptic ulcer (Salim, 1990). Therefore, it has recently been proposed to study the possible antioxidant activity of antiulcerogenic extracts, fractions and/or compounds for the elucidation of their mode of action (Yoshikawa et al., 1993).

In the present study, the hydroxyl radical scavenging ability of the extracts, fractions and constituents obtained through activity-guided fractionation were studied by using in vitro electron spin resonance (ESR) spectrometry technique, in order to elucidate the role of antioxidant principles in the potent antiulcerogenic activity of the plant.

Section snippets

Plant material

S. junceum L. flowers were obtained from Foça (İzmir) in May 1997 and a voucher specimen is stored at the Herbarium of the Faculty of Pharmacy, Gazi University (97A001).

Chemicals

5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and xanthine oxidase were purchased from Sigma Chemical Co. (St. Louis, MO), hypoxanthine, diethylenetriaminepentaacetic acid (DETAPAC) were obtained from Wako Pure Chemical Industry (Osaka, Japan). Other chemicals used for the fractionation and isolation processes were as follows: Diaion

Results and discussion

Reactive free radical species were recently suggested to contribute to cellular injury in the pathogenesis of various gastric mucosal damages (Yoshikawa et al., 1993). Among these, the hydroxyl radical is known as an extremely highly reactive oxygen species, which reacts rapidly with biological materials, causing oxidative damage. ESR spin trapping technique using DMPO has been employed for the indirect and specific detection of hydroxyl radicals and quantification of free radical scavenging

Acknowledgements

The first author, E.Y., acknowledges receiving a grant from Fujii-Otsuka International Fund to the study in Japan.

References (14)

A Arora et al.
Structure-activity relationships for antioxidant activities of a series of flavonoids in a liposomal system
Free Radical Biology and Medicine
(1998)
N Cotelle et al.
Antioxidant properties of hydroxy-flavones
Free Radical Biology and Medicine
(1996)
M.A Perry et al.
Role of oxygen radicals in ischemic-induced lesions in the cat stomach
Gastroenterology
(1986)
C.A Rice-Evans et al.
Structure-antioxidant activity relationships of flavonoids and phenolic acids
Free Radical Biology and Medicine
(1996)
A.S Salim
Oxygen-derived free radicals and the prevention of duodenal ulcer relapse: a new approach
American Journal of Medical Sciences
(1990)
S.A.B.E van Acker et al.
Structural aspects of antioxidant activity of flavonoids
Free Radical Biology and Medicine
(1996)
E Yeşilada et al.
A saponin with anti-ulcerogenic effect from the flowers of S. junceum
Phytochemistry
(1999)

There are more references available in the full text version of this article.

Cited by (66)

Chemical constituents from Diplopterygium chinense (Rosenstock) De Vol and their chemotaxonomic significance
2023, Biochemical Systematics and Ecology
A phytochemical study on the aerial parts of Diplopterygium chinense (Rosenstock) De Vol yielded fifteen compounds consisting of five steroids (1–3, 8, and 9), two diterpenoids (4 and 5), two triterpenoids (6 and 7), three flavone glycosides (10, 11, and 13), and three flavonol glycosides (12, 14, and 15). Their structures were identified according to NMR, MS, ORD, and CD data. This is the first study on the chemical constituents of D. chinense. Compounds 1–4, 6–8, 11, 12, 14, and 15 were first found from the genus Diplopterygium, while compounds 7, 11, 14, and 15 were first reported from the family Gleicheniaceae. Furthermore, the chemotaxonomic significance of these compounds was discussed.
Comparative antioxidant, antibacterial, and GC-MS analysis of methanol extract's fractions and isolation of luteolin from leaves of trans-Himalayan Codonopsis clematidea
2020, Industrial Crops and Products
Codonopsis clematidea, is a trans-Himalayan cold desert medicinal plant species, traditionally used by local tribal communities for the cure of various ailments. On this basis in present study the phytochemical composition and biological potential (antibacterial and antioxidant) of various fractions (n-hexane, chloroform, ethyl acetate, and n-butanol) of methanol extract obtained by soxhlet extraction technique from leaves of Codonopsis clematidea were investigated. Different fractions were accessed for their yield percentage, total polyphenol content (TPC), total flavonoid content (TFC), antioxidant potential (DPPH - 1,1-diphenyl-2-picrylhydrazyl; ABTS - 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), reducing power assay (FRAP - ferric reducing antioxidant power activity), and antibacterial activity (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli). Further, different fractions were also subjected to gas chromatography - mass spectrometry (GC–MS) analysis in order to assess the composition of various volatile phytoconstituents. In antioxidant assays, ethyl acetate fractions (EF) showed the strongest DPPH and ABTS antioxidant (IC₅₀: 0.082 and 0.004 mg/mL) and FRAP potential, which significantly correlate to its high content of polyphenolics and flavonoids. However, EF has also demonstrated similar response in antibacterial activity. Further, luteolin was isolated and identified in EF using chromatography techniques and nuclear magnetic resonance (NMR) spectroscopy. The present results support the possible use of the ethyl acetate fraction from Codonopsis clematidea in food, pharmaceutical, and cosmetic industries.
Hydroalcoholic extract of Spartium junceum L. flowers inhibits growth and melanogenesis in B16-F10 cells by inducing senescence
2018, Phytomedicine
Ultraviolet light exposure generates, in human tissues, radical species, which represent the main cause of photo-aging, DNA damage and skin cancer onset. On the other hand, Mediterranean plants, being continuously subjected to high solar radiation levels, are naturally adapted to take on this type of abiotic stress, thanks to the production of antioxidant secondary metabolites. For these reasons, several plant extracts were documented to be excellent antineoplastic drugs.
We investigated the potential antitumor activity of the flower extract obtained by Spartium junceum L., a Mediterranean shrub, correlating it with the plant metabolic profile.
After selecting the best extraction method to obtain as more secondary metabolites as possible from S. junceum flowers, we characterized the extract metabolic content. Then, by in vitro analyses, the antioxidant profile and the antineoplastic activity on B16-F10 murine melanoma cell of our extract were investigated.
Spectrophotometric assays, HPLC-DAD and GC-MS analyses provided us information about flower extract composition and antioxidant activity. MTT assay and Trypan Blue exclusion test were performed to assess the extract toxicity and the viability, after treatments, of B16-F10 cancer cells and of C2C12 murine myoblasts. In vitro experiments (i.e. cytofluorimetry, protein analysis and qPCR) allowed us to analyze the effect of the plant extract on B16-F10 cell redox state, melanogenesis and cell cycle. Senescence induction was investigated by using a specific kit.
We observed that the hydroalcoholic extract of S. junceum flowers (HFE) strongly inhibited B16-F10 murine melanoma cell proliferation, while just a feeble effect was observed on C2C12 murine myoblasts. Moreover, we found that HFE exerted a pro-oxidant activity on melanoma cells, inhibited melanogenesis and caused cell cycle arrest in G2/M phase, inducing senescence. These anti-cancer properties of HFE could be related to the rich metabolic profile of the extract that we characterized by HPLC-DAD and GC-MS analyses.
This evidence suggests that S. junceum phytocomplex can be used as a selective, nontoxic, economic and easily available anticancer drug.
Anatomical and chemical characteristics of leaves and branches of Juniperus deppeana var. deppeana (Cupressaceae): A potential source of raw materials for the perfume and sweet candies industries
2018, Industrial Crops and Products
Citation Excerpt :
All these compounds contain highly double-bonds conjugation in their chemical structures, which are reported to be responsible, in part, for the antioxidant and anti-inflammatory activities in several plants as well as the antimicrobial properties. This biological potential is due to the great capacity of electron delocalization of these molecules (Koike et al., 2015; Rice et al., 1996; Yesilada et al., 2000). More than twenty terpenes were identified in the essential oils of leaves and branches on the basis of their retention time and mass spectrometry data and some of them corroborated by co-analysis with commercial standards (Table 3).
In North America, the genus Juniperus has a high commercial value because of the natural products derived from its wood, bark and leaves. In Mexico, the species of this genus are commonly known as junípero, sabino, enebro and táscate and they could represent potential sources of valuable products. To the best of our knowledge, this is the first report dealing with the anatomy and the phytochemistry of the endemic species Juniperus deppeana var. deppeana. From the first viewpoint, the resin canals are very large and they occupy one third of leaf volume. In the leaves, they are round, large and turgent, whereas in the branches, they merge and form large, elliptical cavities. The origin of resin ducts in branches is either the cortex or the phloem. They never form in the xylem. The phytochemical analysis of the crude extracts and essential oil of the leaves and branches showed that J. deppeana is a rich source of important polyphenolics and essences and might have the potential to be used in formulations containing this plant with applications in different areas, such as cosmetics, food and health care industries.
Isolation and structure elucidation of two new antioxidant flavonoid glycosides and fatty acid composition in Hedysarum carnosum Desf.
2016, Industrial Crops and Products
The present work constitutes a contribution to the phytochemical investigation of Hedysarum carnosum aiming to search for novel bioactives secondary metabolites. In this context, the fractionation of the butanolic extract of H. carnosum flowers afforded two new flavonoid glycosides quercetin 3-O-β-d-glucopyranosyl-(1 → 2)-α-l-rhamnopyranosyl-(1 → 6)-α-l-rhamnopyranoside 1 and kaempferol 3-O-β-d-glucopyranosyl-(1 → 2)-α-l-rhamnopyranosyl-(1 → 6)-α-l-rhamnopyranoside 2 together with a the known rutin 3 indicated for the first time in this species. Their structures were determined on the basis of spectroscopic evidences (UV, HRMS and NMR) and comparison with literature. The fatty acid composition of the hexane extract of this species was also investigated by using GC–MS. The n-butanol extract and the isolates were assessed for their antioxidant potential using DPPH and ABTS⁺ scavenging assays and ferric reducing power (FRAP). The n-butanol extract and its fractions exhibited efficient radical scavenging activity with IC₅₀ values ranging from 16 to 37 μg/mL for the DPPH assay. The obtained results suggest that the isolated compounds 1 and 3 could be promising antioxidant agents.
Gastroprotective effect of Cymbopogon citratus infusion on acute ethanol-induced gastric lesions in rats
2015, Journal of Ethnopharmacology
Treatment of gastric ulcers with medicinal plants is quite common in traditional medicine worldwide. Cymbopogon citratus (DC) Stapf. leaves infusion has been used in folk medicine of many tropical and subtropical regions to treat gastric disturbances. The aim of this study was to assess the potential gastroprotective activity of an essential oil-free infusion from C. citratus leaves in acute gastric lesions induced by ethanol in rat.
The study was performed on adult male Wistar rats (234.0±22.7 g) fasted for 24 h but with free access to water. The extract was given orally before (prevention) or after (treatment) intragastric administration of absolute ethanol. Effects of dose (28 or 56 mg/kg of body weight) and time of contact of the extract with gastric mucosa (1 or 2 h) were also assessed. Animals were sacrificed, being the stomachs removed and the lesions were assessed by macroscopic observation and histopathology.
C. citratus extract, given orally before or after ethanol, significantly (P<0.01) reduced gastric mucosal injury compared with control group (vehicle+ethanol). The effect does not appear to be dose-dependent. Results also suggested that the extract is more effective when the time of contact with gastric mucosa increases.
The results of this assay confirm the gastroprotective activity of C. citratus extract on experimental gastric lesions induced by ethanol, contributing for the pharmacological validation of its traditional use.

View all citing articles on Scopus

View full text

Article preview

Journal of Ethnopharmacology

Abstract

Introduction

Section snippets

Plant material

Chemicals

Results and discussion

Acknowledgements

References (14)

Structure-activity relationships for antioxidant activities of a series of flavonoids in a liposomal system

Free Radical Biology and Medicine

Antioxidant properties of hydroxy-flavones

Free Radical Biology and Medicine

Role of oxygen radicals in ischemic-induced lesions in the cat stomach

Gastroenterology

Structure-antioxidant activity relationships of flavonoids and phenolic acids

Free Radical Biology and Medicine

Oxygen-derived free radicals and the prevention of duodenal ulcer relapse: a new approach

American Journal of Medical Sciences

Structural aspects of antioxidant activity of flavonoids

Free Radical Biology and Medicine

A saponin with anti-ulcerogenic effect from the flowers of S. junceum

Phytochemistry

Cited by (66)

Chemical constituents from Diplopterygium chinense (Rosenstock) De Vol and their chemotaxonomic significance

Comparative antioxidant, antibacterial, and GC-MS analysis of methanol extract's fractions and isolation of luteolin from leaves of trans-Himalayan Codonopsis clematidea

Hydroalcoholic extract of Spartium junceum L. flowers inhibits growth and melanogenesis in B16-F10 cells by inducing senescence

Anatomical and chemical characteristics of leaves and branches of Juniperus deppeana var. deppeana (Cupressaceae): A potential source of raw materials for the perfume and sweet candies industries

Isolation and structure elucidation of two new antioxidant flavonoid glycosides and fatty acid composition in Hedysarum carnosum Desf.

Gastroprotective effect of Cymbopogon citratus infusion on acute ethanol-induced gastric lesions in rats