1 Institute of Pharmacy and 2 Institute of Organic and Macromolecular Chemistry, Friedrich-Schiller-University, Jena, Germany
3 Research Center on Entomogenous Fungi, Anhui Agricultural University, Anhui, China
Insect pathogenic fungi are a taxonomically diverse group of highly specialized fungi which have been largely ignored up till now as a source of bioactive metabolites, despite the fact that some species have found use as biocontrol agents and certain Cordyceps species are highly regarded in Traditional Chinese Medicine. We have recently embarked on the exploration of entomogenous deuteromycetes for novel bioactive metabolites. In a screening for activity on various CNS related targets, some 32 strains belonging to 10 different genera were grown in SDAY and modified Sabouraud media for 21days at 25 ºC as shake and still cultures. Mycelial and broth extracts were obtained after separation of biomass, freeze drying and extraction with MeOH. A total of 256 extracts were thus obtained and screened for radical scavenging activity (2), MAO inhibition, and for neuritogenic properties in PC12 cells. The latter cell line is known to respond to nerve growth factor (NGF) and other neuritogenic compounds by differentiation into a neuron-like morphology with elongated outgrowths (3). In this assay mycelial extracts of several Paecilomyces species were found to induce neurite sprouting at a concentration of 100µg/ml.
Activity guided fractionation of a mycelial extract from Paecilomyces militaris RCEF 0095 by HSCCC, preparative RP chromatography and gel filtration on Sephadex LH 20 afforded a novel metabolite 1 and other structurally related pyridone alkaloids. Their structures were established by ESI MS, HR EIMS, and extensive 1 and 2D homo- and heteronuclear NMR spectroscopy.
1
Compound 1 induced neuron-like outgrowth in PC12 cells at 3 µM concentration. No significant cytotoxicity was found at this concentration on PC12 and 1321N1 astrocytoma cells.
Biogenetically related pyridone antifungals have been reported from other fungal genera such as Beauveria (1), Agiospora and Penicillium , but neuritogenic properties were not previously discovered for this compound class.
[1] Khachatourians GG (1996). Biochemistry and molecular biology of entomopathogenic fungi. In: Howard DE, Miller JD, eds. The Mycota VI, Human and Animal Relationships. Berlin: Springer, pp 331 - 363
[2] Hu F, Schmidt K, Stoyanova S, Li Z, Gräfe U, Hamburger M (2001). Radical scavengers from the entomogenous deuteromycete Beauveria amorpha, Planta Medica, in press.
[3] Greene L, Rukenstein A (1989). The quantitative bioassay of NGF with PC12 cells. In: Rush R, ed. Nerve Growth Factors, Wiley: New York, pp 139 147
1Institut des Régions Arides, Laboratoire eau-sol-végétaux, 4119 Medenine, Tunisia;
2Faculté de Pharmacie, Laboratoire de Pharmacognosie, 5000 Monastir, Tunisia;
3Faculté de Médecine, Département de Biochimie, Laboratoire USCR/SM, 5010 Monastir, Tunisia.
Essential oils were isolated by hydrodistillation from Artemisia herba alba, Thymus capitatus, Rosmarinus officinalis and Juniperus phoenicea , growing spontaneously in the south-east of Tunisia and usually used in folk medicine. The essential oil yield was respectively 0.65, 2.75, 2.50 and 0.70%. These oils were investigated by GC-MS. Artemisia herba alba oil was mainly composed of a-thujone (44%), sabinyle acetate (17%) and b-thujone (10%). The major components of Thymus capitatus oil were thymol (66%), p-cymene (6%) and sabinene (4%). The most abundant components of the oil of Rosmarinus officinalis were 1,8-cineole (27%), camphene (23%) and b-pinene (8%). The oil of Juniperus phoenicea was mainly composed of a-pinene (68%) and p-cymene (6%).
Chemistry of Medicinal Plants Lab., National Research Center, Dokki, Cairo, Egypt
The genus Acacia belongs to Leguminosae family. Information about the constituents of the studied species Acacia ehrenbergiana is very limited The phytochemical screening (1) of A. ehrenbergiana showed the presence of hydrocarbons, fatty acids, fatty alcohols, sterols, terpenes, flavonoids, anthraquinons, tannins and saponins.
The n-hexane extract of the aerial Parts (450 g) of the plant was dissolved in hot acetone. The investigation of the insoluble acetone fraction revealed the presence of different fatty alcohols. Cosanol, tricosanol, tetracosanol, pentacosanol, hexacosanol, octacosanol, tricontanol and dotricontanol could be identified by means of GLC (2).
The acetone soluble fraction was subjected to column chromatography eluted with gradient benzene in hexane. The fraction eluted with 50% benzene in hexane shows the presence of three main sterols were identified by GLC (3), stigamasterol; campesterol and cholesterol with different percentages.
1 Kg of A. ehrenbergiana was extracted with 80% acetone. The aqueous extract after evaporation of the aceton was partitionated with petroleum ether followed by ethyl acetate. The ethyl acetate fraction was subjected to sephadex LH-20 column chromatography eluted with 96% ethyl alcohol. Six main collective fractions were collected.
Diosmetin, taxifolin, kaempferol, myricytin and apigenin were separated and identified by spectreal analyses (4). Also gallic acid, methyl gallate, flavan-3-ol-gallate and gallocatechin-3-gallate were identified.
Wall, M.E.; Krieder, M.M.; Krewson, C.F.; Eddy, C.R.; William, J.; Carel, D.S. and Centry, H.S. J. Amer. Pharm. Assoc. 43 ,1, (1954).
Varian / model 3700 Gas chromatograph, 15% DEGS on Chromosorb (W/AW), 80/100,2m, SSt, 0.25 i.d., 240°C, 280°C, 70-190°C, 55 min., 2°C/4 min., Helium, 30 ml/min.
Varian/ Finnigan mat SSQ 7000 (USA), DB-5 fused silica, 30 ml. Length, 0.25 mm i.d. and 0.25 mm thickness, 230°C, 27°C, 70-270°C, 10°C min. for 20 min, then 270°C for 25 min., Helium, 30 ml/min.
Mabry, T.J.; Markham, K.R. and Thomas, M.B., Cited in " The systematic Identification of Flavonoids", Springer - Verlag, Berlin , (1970).
1Department of Applied BioSciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland
2Department of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, TR-06100 Ankara, Turkey
In the flora of Turkey the genus Ajuga L. is represented by 11 species [1] some of which are traditionally used in wound healing, as diuretic as well as against diarrhea and high fever [2]. There have been many phytochemical investigations on Ajuga species focusing mainly on the isolation of phytoecdysteroids and diterpenes. However, there was only one report on Ajuga salicifolia (L.) Schreber, concerning the isolation of a diterpenoid [3]. Previously we have reported the isolation and structure elucidation of ionone and iridoid glycosides from the aerial parts of A. salicifolia, collected from Ankara [4].
Further chromatographic investigations on the methanol extract of the title plant by VLC (RP-18), subsequent CC (silica gel), and HPLC (RP-18) resulted in the isolation of three novel sterol glycosides (1- 3).
The structures of the compounds were elucidated by one and two dimensional NMR techniques (1H, 13C, 13C/DEPT, DQF-COSY, HMBC, HSQC, HSQC-TOCSY, NOESY, ROESY) and high resolution mass spectrometry.
Davis, P. H., Flora of Turkey and the East Aegean Islands, Edinburg, 1982, p.7.
Baytop, T., Therapy with Medicinal Plants (Past and Present), Istanbul University Publications, Istanbul, 1984, p.298, p.416.
Bozov, P. I., Papanov, G. Y., Malakov, P. Y., De La Torre, M. C., Rodriguez, B. (1993) Phytochemistry 34: 1173-1175.
Akbay, P., Çalis, I., Heilmann, J., Sticher, O. (2000) Ionone and Iridoid Glucosides from Ajuga salicifolia, Abstract No. P1A/01, 48th Annual Congress of the Society for Medicinal Plant Research, September 3-7, Zurich, Switzerland.
Department of Chemistry, University of Bahrain, P.O.Box 32038
Cordia myxa L. (Boraginaceae) is one of the traditional fruits growing in Bahrain. It is widely used in Bahrain for treating whooping cough, as a laxative, vermicide and diuretic. The phytochemical screening of both the leaves and the fruits showed the presence of alkaloids, coumarins, flavonoids, saponins, and terpenes and / or sterols. The ethanolic extracts prepared from either fruits or leaves were checked for their antimicrobial (against Gram-positive and Gram-negative bacteria) and antioxidant activities. The extracts showed remarkable antioxidant activities but no antimicrobial activity in two different concentrations. The fresh leaves were analyzed by atomic absorption spectroscopy for toxic cadmium (Cd), mercury (Hg), and lead (Pb) and for the biologically essential metals iron (Fe), copper (Cu) and zinc (Zn). The mean concentration of Cd, Hg, Pb, Fe. Cu and Zn were 0.26, 1.64, 0.085, 117.0, 6.0, and 18.3 mg/kg, respectively.
* Department of Pharmacognosy, Faculty of Pharmacy, Assiut University, Assiut 71526, Egypt
**Department of Pharmacognosy, Faculty of Pharmacy, Kasr El-Aini Cairo University, Cairo11562, Egypt
Pulicaria crispa (Forsk.) (syn. Francoeuria crispa Benth et Hook, family Asteraceae, Tribe Inuleae) is an important aromatic, bushy desert plant growing wild in Aswan region. Locally it is known as kut-kat, Sabat, Francoeuria , Tagar, Ra' ra. Activity-guided fractionation of the alcoholic extract of Pulicaria crispa lead to the isolation of four phenolic compounds using combined column chromatography and preparative TLC (silica gel) from the light petrolium-ether (1:1) fraction.
The isolated compounds were identified as quercetin 3,7-dimethyl ether, p-hydroxybenzoic acid, 3,4-dihydoxy-benzoic acid and 4-hydroxy-3-methoxy-benzoic acid based on their spectroscopic data.
The HIV-1 protease inhibitory activity of the light petroleum-ether (1:1) fraction as well as the isolated compounds was determined by incubating them in a reaction mixture containing protase enzyme (PR) and substrate His-Lys-Ala-Arg-Val-Leu-(pNO 2-Phe)-Glu-Ala-NIe-Ser-NH2 at pH 5 to perform protolytic cleavage reaction.
The products of proteolysis were analysed by HPLC. The isolated compounds quercitin 3,7-dimethyl ether, p-hydroxybenzoic acid, 3,4-dihydoxy-benzoic acid and 4-hydroxy-3-methoxy-benzoic acid showed IC50 of 0.25 mM, > 0.60 mM, 0.325 mM and 0.55 mM, respectively.
1Imperial College of Science, Technology and Medicine, Dept of Biological Chemistry, SAF Building, Exhibition Road, South Kensington, London SW7 2AZ, United Kingdom
2Kings College London, Department of Pharmacy, Franklin-Wilkins Building, 150 Stamford Street, Waterloo, London SE1 8WA, United Kingdom
3Oxford Natural Products plc, 1 St Giles, Oxford OX1 3JS, United Kingdom
Although utilisation of natural products in the western world has grown rapidly in the last decade, the formulation of such products is not regulated as stringently as conventional pharmaceuticals. Consequently, ostensibly identical raw materials and similar products that are marketed by different manufacturers can display variation both in the quality and quantity of active ingredients. By applying chemometric analysis to extracts of such materials, it is possible to determine statistically significant parameters with which to assess and control production. Further, with increasing interest in the development of rational approaches to the characterisation of plant extracts respecting the observation that the biological activity of the total extract is often a combined effect due to multiple components acting by several mechanisms, the combination of NMR spectroscopy and chemometric analysis offers a powerful approach to total chemical analysis (manuscript submitted for publication).
In this study, high field 1H NMR spectroscopic analysis has been used in conjunction with principal components analysis to characterise the chemical composition of extracts of a range of samples of Paeonia lactiflora, used in traditional Chinese medicine for the treatment of dysmenorrhoea. The work indicates that not only can such multivariate data analysis be employed for the discrimination between P. lactiflora samples from different sources, but, in addition, the utilisation of predictive models allows the classification of previously unanalysed samples. This provides an indication as to whether or not a particular sample is likely to exhibit similar effects and potencies to those of extracts already characterised.
These results demonstrate that the combination of NMR spectroscopy and chemometric analyses of natural product analytical data can provide rapid characterisation and determination of differences in chemical composition, with obvious implications for screening in the phytochemical arena and for the quality control of herbal products.
Pharmacognosy and Chemistry of Medicinal Plants Dept., National Research Center, Dokki, Cairo, Egypt.
Algae are important source of therapeutically useful substances (1). Bioactive diterpenes of parguerene and isoparguerene carbon skeleton have been isolated from Laurencia spp. (2,3) and Corallina officinalis L. (4). Ether fraction of methanol extract of fresh alga Jania rubens (L) Lamx. collected from the Red Sea coast at Hurghada, Egypt was fractionated by silica gel column chromatography. Seven brominated diterpenes of the parguerene and isoparguerene series have been isolated for the first time from this alga. The spectroscopic features of these compounds proved to be isoparguerol (1), isoparguerol-16- acetate (2), isoparguerol-7,16-diacetate (3), parguerol-16-acetate (4), parguerol- 7,16-diacetate (5), deoxyparguerol (6) and deoxyparguerol-7- acetate (7).
The antitumour activities of the isolated compounds have been examined in vitro using Ehrlich ascites carcinoma cell cultures. The results revealed that ibrominated diterpenes described here have a marked antitumour activity. It seems as if isoparguerol derivatives are slightly more effective than parguerol derivatives, deoxyparguerol derivatives showing the weakest activity. Moreover, the data imply that the cytotoxicity of these compounds are dependent on the number of acetoxy groups.
1. Nagwa E Awad (2000) Phytotherapy Research, 14, 641.
2. Takeda, S; Kurosawa, E; Komiyama, K. and Suzuki,T.(1990) Bull. Chem. Soc. Jpn., 63, 3066.
3. Rochfort, S.J. and Capon, R.J. (1996) Aust. J. Chem., 49, 19.
4. Nagwa E Awad; Saleh, M.M.; Selim, M A and Matloub, A.A. (1999) Egypt. J.Pharm.Sci.40(2)
Pharmacognosy and Chemistry of Medicinal Plants Dept., National Research Center, Dokki, Cairo, Egypt.
Coralline algae (Corallina officinalis L. and Jania rubens (L) Lamx.) have been reported to have many biological activities (1,2). Volatile constituents, fatty alcohols and hydrocarbons fraction of Jania rubens (L) Lamx. have been isolated for the first time and identified by GC/MS. Fifty volatile constituents which constitute 92.88% of the total volatiles and 25 components of hydrocarbon fraction which represent 88.18% were identified. The fatty alcohols fraction consists of n-tridecanol (38.35%), 2-tetradecanol (12.71%), n-pentadecanol (18.38%) and n-octadecanol (30.56%).
The antibacterial activities of these four compounds were examined applying the disc diffusion technique (3). Different concentrations of the isolated compounds (50, 100 and 200 µg/disc) were tested against Bacillus subtilis, Streptococcus lactis, Pseudomonas putidu and E. coli. The isolated constituents exhibited marked antimicrobial activities against the tested organisms.
1. Nagwa E Awad; Selim, M.A.; Saleh, M. M. and Matloub, A. Phytotherapy Research, in press
2. Soliman, F.M.; Tohamy. S.F; Fathy, M M; Ramadan, A; Afify, N A. and Sanad, O A (1994) Drug Res. 21 (1-2), 165.
3. Gnanamanickam and Mansfield J W (1981) Phytochemistry 20, 997.
1Institut für Organische Chemie, Universität Hamburg, Martin-Luther-King-Platz 6, D-20146 Hamburg; Germany
2Frohme-Apotheke, Frohmestr. 14, D-22457 Hamburg, Germany
Olibanum belongs to the group of oleogum resins that exudes from incisions in the bark of trees of the genus Boswellia (Burseraceae). T he trees yielding the resin are native to Ethiopia, Somalia, the Arabian peninsula and India. Its use as an anti-inflammatory and antiseptic agent are well known in folk medicine [1].
Our previous studies showed that B. carterii and B. serrata revealed characteristic differences in their essential oils. The inspection of the GC-MS data with the comparison of mass spectra and retention indices with our database showed that both species contain many commonly found mono- and sesquiterpenes in which octylacetate and a-pinene are the major components for B. carterii and B. serrata, respectively. The presence of the diterpene derivatives incensole, incensole acetate as well as cembrene, cembrene A, cembrene C and verticilla-4(20),7,11-triene for B. carterii and cembrenol (cembra-3,7,11-triene-1-ol) for B. serrata serve as diagnostic markers for these species [2].
In this study we would like to present results for B. frereana since it shows differences in its GC-MS data and HPTLC fingerprints in comparison to the above mentioned species. Several isomeric dimers of a-phellandrene (Fig.1) were found for the first time as natural products in these species. Their relative configurations were identified by 1- and 2-D NMR spectroscopy.
In addition to these, headspace SPME studies will be presented for the three species which are found to be comparable in their composition with the corresponding hydrodistillation products.
|
Fig. 1. Dimer of a-phellandrene |
1. Martinetz, D., Lohs, K., Janzen, J., Weihrauch und Myrrhe, 1988. 2. Basar, S., Koch, A., König, W. A., A verticillane type diterpene from Boswellia carterii essential oil, Flav. and Frag. J., 2001, in press. |
1School of Pharmacy and Biomedical Sciences, University of Portsmouth, U. K.;
2Department of Chemistry, United Arab Emirates University, Al-Ain, United Arab Emirates;
3Environmental Research and Wildlife Development Agency, Abu Dhabi, United Arab Emirates;
4Institute of Pharmacognosy, Albert Szent-Gyorgyi Medical and Phannaceutical Centre, University of Szeged, Szeged, Hungary
5Institute of Ecology and Botany ofthe Hungarian Academy of Sciences, Vácrátát, Hungary
From Haplophyllum tuberculatum (Forssk.) A. Juss. (Rutaceae) grown in Egypt, essential oils were obtained from plants collected in the spring and summer (l). The composition of the two oils varied considerably. Monoterpene hydrocarbons were the major components of both, but in the spring sample, the percentage of these compounds was double that of the summer sample. d -3-carene was the major constituent of both samples and myrcene was also a significant component. The composition of the essential oil of air-dried parts of H. tuberculatum grown in Iran has also been reported (2). Monoterpene hydrocarbons again predominated, but the major compounds were limonene (27.3 %), a -pinene (21.9 %), myrcene (6.9 %) and camphene (5.2 %). The proportion of d-3-carene was only 3.3 %.
In this report, the composition of the essential oil isolated from H. tuberculatum collected from the United Arab Emirates (UAE) was studied. Plant material was collected from Khor Fakkan, UAE, in May 1997 and April 1998. The essential oil was isolated from the fresh aerial parts, including flowers, by hydrodistillation for 5 h according to the European Pharmacopoeia method (yields: May 0.08 %; April 0.04 %). The collected oil was dried using anhydrous sodium sulphate. The compositions of the essential oils were determined using GC/FID and GC/MS techniques. The constituents were identified by comparing their retention times and Kovats indices with those of authentic reference compounds, by comparison with published MS data and from Computer library searches.
In the oil of H. tuberculatum distilled from the May 1997 collection, 23 components were characterised and 27 from the April 1998 sample. In both, monoterpenoids predominated, as was the case with the Egyptian (l) and Iranian (2) oils. Many differences were found between the May 1997 and April 1998 oils. In the May collection, a-phellandrene was the major component (32.9 %), whereas this formed only 4.8 % of the April sample. The most abundant compounds in the April oil were linalool (15.0 %) and linalyl acetate (10.6 %), whereas in the May sample, linalool formed only 3.4 % of the total oil and linalyl acetate was not detected. The sesquiterpenoid b-caryophyllene was a major component of both the May (12.8 %) and April (9.7 %) oils. d-3-carene, which was reported as the major constituent of the Egyptian oils (l), comprised only 6.0 % and 1.1%, respectively of the May 1997 and April 1998 UAE samples and 3.3 % of the Iranian oil. Limonene and a -pinene, the predominant components of the Iranian oil, were only minor constituents of the UAE and Egyptian oils.
1. Brunke, E. J., Hammerschmidt, M. A., Abd El-Kawy, E. A., El-Kashoury, A., Soliman, F. M. Herba Hungarica 1991, 30, 34-39.
2. Yari, M., Masoudi, S., Rustaiyan, A. J. Essent. Oil Res. 2000, 12, 69-70.
Institute of Pharmaceutical Biology and Phytochemistry, Hittorfstr. 56, 48149 Münster, Germany
Harpagophytum procumbens DC (Pedaliaceae) is a perennial herbaceous plant that grows in the South-West area of Africa. In Europe, extracts of the secondary storage roots of H. procumbens are used for the treatment of rheumatic and gastro-intestinal disorders. H. procumbens is known for the occurrence of iridoid and phenylethanoid glucosides.
The aim of this study was to investigate an aqueous extract prepared from the storage roots in order to improve the knowledge about its phytochemical composition and to identify compounds with antirheumatic properties.
We isolated harpagoside (1), 8-p-coumaroylharpagide (2), a 5-deoxyharpagoside (3 ), pagoside (4), 8-feruloylharpagide (5), acteoside (6 ), isoacteoside (7), 6-O-acetylacteoside (8), cinnamic acid (9) and caffeic acid (10) by means of CC on Celite ä, MLCCC and finally HPLC on RP 18.
Compounds 1, 2, 6, and 7 are already known as constituents of Harpagophyti radix. Compounds 3, 4, 5, 7, 9 and 10 were not yet detected in H. procumbens. Compound 4, for which we propose the name pagoside, and 8 were isolated for the first time from a natural source. The structures were elucidated by ESI-MS, 1D-and 2D-NMR measurements.
HPLC was used to quantify 1-10 in aqueous extracts of the crude drug and in aqueous solutions.
1Department of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, TR-06100 Ankara, Turkey
2Department of Medicinal Chemistry, College of Pharmacy, University of Utah, Salt Lake City, Utah 84112, USA
3Department of Applied BioSciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich, CH-8057 Zürich, Switzerland
From the aerial parts of Putoria calabarica (Rubiaceae), we recently published several iridoid and lignan glycosides as well as flavonoid glycosides with significant antioxidant activity [1]. Further investigations of the same plant using a combination of Silica CC, Sephadex LH-20 CC and C-18 MPLC resulted in the isolation of two new (1, 2) and two known (3, 4) lucidin-type anthraquinone glycosides. On the basis of spectroscopic methods (1D- and 2D NMR, MS, UV, IR), the structures of compounds 1 and 2 were elucidated as 5,6-dihydroxy-1-methoxy-2-hydroxymethyl-9,10-anthraquinone-3- O-b -glucopyranoside and 6-hydroxy-1-methoxy-2-hydroxymethyl-9,10-anthraquinone-3- O-b -glucopyranoside, respectively. We propose the trivial names of putorinosides A and B for compounds 1 and 2. Compounds 3 and 4 were identified as lucidin-3-O-b-glycopyranoside and lucidin-3-O-b-primeveroside.
Institute of Pharmacy, Department of Pharmaceutical Biology, University of Kiel, Gutenbergstr. 76, D- 24118 Kiel, Germany
A typical feature of leaves and flowers of Alcea rosea is the presence of mucilage, which is located in idioblasts, large cavities or specialized epidermal cells (1). Main components of these mucilages have been found to be of rhamnogalacturonan type (2). Aim of the present study was to establish cell cultures of Alcea rosea in order to isolate dominant polysaccharides produced in cell culture and to compare them with those of genuine mucilages.
An arabinogalactan-protein (AGP) from suspension culture medium was isolated by precipitation with ethanol, followed by precipitation with b-glucosyl Yariv's reagent. It revealed characteristic features of AGPs like a high amount of polysaccharide and a small protein moiety with the main amino acids serine, alanine, hydroxyproline, threonine, glutamine, asparagine and arginine and was quite similar to an AGP from cell culture of Malva sylvestris (3). The molecular weight was estimated to be 1.4 x 106 Da. Linkage analyses showed that the AGP is composed of a highly branched core polysaccharide of 3-, 6-, and 3,6-linked Galp residues with terminal Araf, GlcAp and Galp. Estimation of the AGP content in suspension culture media turned out to be possible by photometric quantification with Yariv´s reagent.
Pharmacological investigations have shown immunomodulatory activities of arabinogalactan-type polysaccharides as e.g. activation of macrophages, activation of complement system and stimulation of NK cell cytotoxicity (4). These results are a hint that isolation of AGPs from the spent medium of suspension-cultured cells in combination with their pharmacological testing might be a challenging task for the future.
Classen, B., Dissertation, Kiel 1997
Classen, B., Blaschek, W. (1998) Planta Med. 64, 640-644.
Classen, B., Blaschek, W. (2001) Planta Med., submitted
Lohmann-Matthes ML, Wagner H (1989) Z. Phytotherapie 10, 52-59
a Institut für Chemie, Universität Hohenheim, Garbenstrasse 30, 70599 Stuttgart, Germany
b Hygiene Institute, Department of Parasitology, Im Neuenheimer Feld 324, 69210 Heidelberg, Germany
c Department of Chemistry, University of Alabama, Huntsville, USA
From Terminalia macroptera (Combretaceae) the following six hydrolyzable tannins: 2,3-O-(S)-hexahydroxydiphenoyl-D-glucose, punicalagin, terflavin A and B, 2-O-galloyl-punicalin, punicacortein C were isolated along with the two novel ellagitannins isoterchebulin (1) and 4,6-O-isoterchebuloyl-D-glucose (2).1
These compounds were evaluated for plant growth regulatory (allelopathic) activity on the radicles and shoots of germinating Radish (Raphanus sativa) var. carnita and Lettuce (Latuca sativa) var great lakes 118.2 The growth regulatory data obtained revealed a structure dependent pattern in the inhibition/stimulation activities on the radicles and stems of the developing seeds. These effects were strongly modulated by Ca2+ and Mg2+ ions.
It is reported that gallic acid methyl ester is active against the malarial parasite, Plasmodium falciparum.3 Since ellagitannins are regarded as biosynthetic derivatives of galloyl glucopyranoses, the following compounds: 2,3-O-(S)-hexahydroxydiphenoyl-D-glucose, 2-O-galloyl-punicalin, isoterchebulin (1), 4,6-O-isoterchebuloyl-D-glucose (2) and punicacortein C were tested against P. falciparum and found to be weak to moderately active as compared to gallic acid methyl ester.4
1. J. Conrad et al., J.Nat.Prod. 2001, 64(3), 294-299.
2. G.O. Buchanan et al., Phytochemistry 2000, 54, 39-45.
3. F.D. Horgen et al., Phytomedicine 1997, 4(4), 353-356.
4. K.T. Andrews et al., Int. J. Parasitol. 2000, 30(6),761-768.
Institute of Pharmacy, Friedrich-Schiller-University, Jena, Germany
Isatis tinctoria L. (woad) is an old European and Chinese dye plant and medicinal herb which has been used since Antiquity in various inflammatory aliments. In a HPLC based bioactivity directed approach we identified the indolo-[2,lb]-quinazoline alkaloid tryptanthrin (1) as the compound responsible for the herbs cyclooxygenase-2 (CQX-2) inhibitory activity [1]. Tryptanthrin is an equally potent inhibitor of 5-lipoxygenase (5-LOX) [2] and thus the first representative of a novel class of dual CQX-2/5-LOX inhibitors.
Tryptanthrin could be reasonably separated by isocratic HPLC on a RP18 column eluted with 40% acetonitrile containing 2% acetic acid. In typical plant samples, the tryptanthrin peaks recorded at the absorption maxima at 254 and 387 nm were too low for reliable integration despite extract concentrations of up to 5 mg ml -1. Considering the versatility, selectivity and sensitivity required in possible future applications, MS detection was a preferred Option over DAD, despite inherent drawbacks such as higher operating cost and bower bong-term stability. ESI-MS conditions were optimized with respect to maximum intensity of the quasimolecular ion at m/z 249 [M+H]+ which was selected for quantitative determination in the single ion mode (SJM). Calibration curves for the range of 1-200 ng were recorded (r2 = 0.9998). The limit for quantitative analysis was 1 ng with a 4 mm column and a 1:5 split. Extraction by ASE (accelerated solvent extraction) [4] combined high tryptanthrin recovery (> 97% with 2 extraction cyles of 5 min) with good reproducibility and possibibity of automation. The between-day R.S.D. ofthe method was 5.57%.
More than 70 Isatis samples were analyzed. Due to the geographical origin ranging from Western Europe to Japan, these specimens were fairly representative of the variability of the species. The tryptanthrin content in leaf samples varied from 0.56 to 16.74 x 10-3% [3].
[1] Danz, H., Stoyanova, S., Wippich, P., Brattström, A., Hamburger, M. (2001) Planta Medica, in press
[2] Danz, H., Stoyanova, S., Thomet,O., Simon, H.-U., Dannhardt, G., Ulbrich, H., Hamburger, H., European Journal of Pharmacology, submitted
[3] Danz, H., Baumann, D., Hamburger, M. (2001) Planta Medica, in press
[4] Benthin, B.; Danz, H.; Hamburger, M. (1999) J. Chromatogr. A, 837: 211-219
Pharmacognosy Department, Faculty of Pharmacy, Assiut University, Assiut, Egypt
The genus Ficus belongs to the
family Moraceae, is predominantly distributed in the tropics and
subtropics. Ficus benghalensis L. is an ornamental
plant named in arabic Teen Benghaly. It is useful in biliousness,
ulcers, vomiting and vaginal complaints. The leaves
are used in
treatment of ulcers while the milky juice is aphrodisiac and tonic,
The root is useful in gonorrhoea and syphilis. From the bark of Ficus
benghalensis L; n-nonadodcane,
b-amyrin,b
-sitosterol, lupene-3-one, lupeol acetate, 24-hydroxy b-sitosterol,
palmitic acid
and palmitoyl glycerol were isolated by different
methods of chromatographic techniques. Besides, four known
flavonoidal compounds were isolated from the leaves.
The isolated compounds were identified by different methods of spectral analysis.
Institute of Pharmaceutical Biology, University of Düsseldorf, Universitätsstr. 1, D-40225 Düsseldorf, Germany
Echinacea products belong to the most popular unspecific immunostimulants. Lipophilic extracts of Echinacea roots, such as E. purpurea and E. angustifolia contain various alkamides, which showed anti-inflammatory and phagocytosis stimulatory activities (1). For chemotaxonomic reasons and to check its immunomodulatory potential we investigated the roots of E. atrorubens Nutt.. We already reported on several isobutylamides and 2-methylbutylamides isolated from E. atrorubens roots (2, 3). We now report on further alkamides from the n-hexane extract of E. atrorubens roots. Pentadeca-2E,9Z-dien-12,14-diynoic acid isobutylamide 1 was isolated by MPLC on RP-18 material. Dodeca-2E,4E-dienoic acid-2-methylbutylamide 2, dodeca-2 E,4E,8Z-trienoic acid-2-methylbutylamide 3, dodeca-2 E,4E,8Z,10E/Z-tetraenoic acid-2-methylbutylamides 4, 5 and the isomers dodeca-2E,10Z/E-dien-8-ynoic acid isobutylamides 6, 7 were obtained by semipreparative HPLC. The compounds were identified by their UV, MS and NMR spectra. Alkamide 1 could be found in E. angustifolia and E. purpurea previously (4, 5). The 10Z-isomer 6 has been already found in E. atrorubens roots (3), but the 10E-isomer 7 is a new compound. Moreover alkamides 2-5 represent new natural compounds. Investigations are in progress to elucidate the pharmacological profile of the alkamides.
Bauer R. (1999) Chemistry, analysis and immunological investigations of Echinacea phytopharmaceuticals. In: Wagner H., Ed. Immunomodulatory Agents from Plants. Basel, Birkhäuser Verlag: 41-88.
Dietz B, Bauer R (2001) Pharmaceutical Biology 39: 11-15.
Dietz B, Bauer R (2000) International Congress and 48 th Annual Meeting of the Society for Medicinal Plant Research (2000), Book of Abstracts, No. P1A/09.
Bauer R, Remiger P, Wagner H (1989) Phytochemistry 28: 505-508.
Bohlmann F, Hoffmann H (1983) Phytochemistry 22: 1173-1175.
Department of Pharmacology, University of Pretoria, Pretoria, South Africa
Acetone extracts of C. microphyllum [also known as C. paniculatum ] leaves have shown substantial antibacterial activity [Eloff 1999] as well as strong antiviral activity against HIV-1 and HIV-2 [Asres et al., 2001]. The extracts are however complex and isolating bioactive compounds is challenging. Only a few extractants have generally been used for isolating antimicrobial compounds from plants and Cowan [1999] concluded that many classes of compounds are commonly obtained in only one solvent. The aim of this study was to investigate whether different solvents could simplify extracts to facilitate the isolation of antibacterial compounds from the complex crude mixture. The solvents were selected to represent a wide range of polarities and selectivity groups [Snyder and Kirkland, 1981].
Intact dried leaves were extracted with acetone and 1 % aqueous sodium bicarbonate and ground leaves were extracted with hexane, carbon tetrachloride, di-isopropylether, ethyl ether, methylene dichloride, tetrahydrofuran, acetone, ethanol, ethyl acetate, methanol and water. The total quantity extracted with each solvent was determined gravimetrically; the complexity of compounds extracted was determined by TLC using different solvent systems and spray reagents. Antibacterial activity of extracts was determined by a microplate serial dilution method [Eloff, 1998]. Total antibacterial activity was calculated by dividing quantity extracted with MIC values [Eloff, 2000]
In contrast to other Combretum spp., sodium bicarbonate gave disappointing results with C. microphyllum leaves. The other solvents extracted from 2.6 to 17.4% of the dry weight. Methanol, methylene dichloride and tetrahydrofuran extracted the largest mass. The chemical composition of the different extracts was remarkably similar with the exception of highly polar [water] and non-polar [hexane] extractants, but the minimum inhibitory concentration for the different extracts varied from 0.01 to 1.25 mg/ml. The average MIC values for the four test organisms were Staphylococcus aureus 0.46 mg/ml Pseudomonas aeruginosa 0.30 mg/ml, Escherichia coli 0.31 mg/ml and Enterococcus faecalis 0.29 mg/ml.
Di-isopropyl ether, ethanol, ethyl ether, acetone and ethyl acetate extracted high antibacterial activity with a lower quantity of other non-active compounds and could be useful for isolating bioactive compounds. The four solvents that extracted the highest total antibacterial activity were: methanol, methylene dichloride, ethanol and acetone. The best solvents belonged to selectivity groups II, III and V [Snyder and Kirkland, 1979], which are good proton-acceptors with a large dipole moment. Because methanol and ethanol extracts also contain highly polar uninteresting compounds, and methylene dichloride is not miscible with water, we decided to use acetone as extractant for isolating antibacterial compounds.
Asres K. Bucar F. Kartnig T. Witvrouw M. Pannecouque C. De Clercq E. [2001]. Phytotherapy Research. 15, 62-69
Eloff J N [1998] Planta Medica 64, 711-714.
Eloff J N [1999]. S.Afr.J.Sci. 95, 148-152.
Eloff J N [2000] S.Afr.J.Sci. 96, 116-118.
Cowan M M [1999]. Clinical Microbiology Reviews 12, 564-582.
Snyder L R and Kirkland J J [1979] Introduction to modern liquid chromatography. John Wiley, New York
* Medicinal and Aromatic Plants Deptartment
** Pharmacognosy Department
National Research Centre, 12311, Dokki, Cairo, Egypt.
Callistemon lanceolatus (Myrtaceae) is an ornamental tree commonly known as bottle brush owing to its flower shape. The leaves contain volatile oil, reported to possess fungicidal and insecticidal activities (l).
Phytochemical investigation was carried on its different extracts for biological evaluation as insecticide and molluscicide from which the chloroformic extract induced higher mortality against cotton leaf worm larvae (80%) in addition to different effects on egg stage and newly hatched snails of Biomphalaria alexandrina; LC50 was 240 ppm . Fractionation of the chlorofomic extract with petroleum ether yielded an amorphous powder identified by spectral tools as b-sitosterol. It showed high toxicity towards the cotton leaf worm with an LC50 of 164 ppm.
From the methanolic fraction of the chloroformic extract, oleanoic acid was isolated by preparative TLC and identified by spectral tools. Biologically it induced pronounced reduction in the total number of eggs, reduced number of clutches and it significantly reduced the newly hatched snails, in addition to reducing shell diameter.
Saponins were also precipitated from the methanolic leaf extract, from which three compounds were isolated. Upon hydrolysis one genin, oleanolic acid, was released together with different sugar moities. The saponin fraction as such only weakly affected the snails' activity; the LC50 was 55O ppm, however the genin released was potent against the snails. The substances isolated from Callistemon leaves are either sterols or triterpenes which mimic insect hormones and hence disturb the fecundity of the pest (2,3).
(1) Mohamed, E.I. and Amer, S.A.A.(1992) Egypt J. Appl.Sci.,7(8):445-456.
(2) Bowers,W.S., Fales, H-M., Thompson, M.J. and Uebol, E.C.(1996) Science,154:1020.
(3) Parkhurst, R., Thomas,D.W. and Skinner,W. (1975). Phytochemistry,12:1437-1442.
* Faculty of Agriculture, Cairo University
** National Research Center, Cairo, Egypt
Zinnia pauciflora a member of the Asteraceae is known to contain some bioactive compounds. In this study some flavonoids were isolated from herb, red and yellow flowers. Some volatile constituents of herb and fatty acids of seed oil were identified and quantified. The compounds cyanidin-3 -gluco-(4-malonyl)-rhamnosoide and cyanidin-3 -(4- malonyl)-arabinoside(1,2) were isolated and purified from red head flowers of Zinnia pauciflora. Apigenin-7-(4-acetyl)-xyloside was isolated and identified(3) from the yellow head flowers as main flavonoids but the main flavonoids of the aerial parts was kaempferol 7-glucorhamnoside. Oxygenated compounds represented the high percentage (89.8%) of total peak area of volatile principles from the herb. The major members of these oxygenated compounds were: phytol (38.3%). Hexandioic dioctyl ester (2 4.9%), hexandecanoic acid (12.4%) and spathulenol (9.4%)(4) .The lipids content was 7.5 % mean fatty acids were linoleic (35%) palmitic (17.3%) and oleic (13.3%)(5)
The crude ethyl acetate extract of aerial parts and yellow flowers was effective against all microorganisms (Bacillus subtilis, Escherichia coli, Aspergillus niger, Fusarium oxvsporium und Candida albicans) .
Low doses of the crude flavonoid fraction prepared from yellow flowers (10 and 90 mg/100g body weight of mail albino rats) did not show any deleterious effect, while high doses (l80 mg/100 g B.wt.) decreased serum creatinine and blood glucose after four weeks. Alanine transaminase, aspartate transaminase and alkaline phosphatase were increased at the high doses which indicated liver dysfunction(6,7)
1. Bride, P.; Loeffer, R.S.; Timberlake, C.F. and Self, R. 1984, Phytochemistry, 23 (12) 2968-69.
2. Takeda, K.; Harborne, J.B. and Self, R. 1986., 25 (6):1337- 1442.
3. Mabry, T.J; Markham, K.R. and Thoma, M.B. 1970. The Systematic identification of Flavonoids Springer Verlag, New York.
4. Adams, R.P. 1989. Identification of Essential Oils by ion trap mass spectroscopy Academic Press, New York.
5. Radami, R. C; Deshpande, G.S. and Shambhag, M.R. 1975 Journal of Oil Technology. (India) 7 (3): 76-77.
6. Abd-El-wahab, S.M.; Wassel. G.M; Ammar N.M. and Hanna, T.1987. Herba Hungarica 26 (1) 27 - 39.
7. Hayat, M.S.; Mohga, S.A.; Yassin, N.A. and Fatma, S. 1995. Bull. Fac. Pharm. Cairo Univ., 33(1): 27-32.
1Chemistry of Tannins and Proteins Dept., National Research Centre, Tahrir Str. , Dokki, Cairo, Egypt
2Institut für Pharmazie, Pharmazeutische Biologie, Universität Leipzig, Johannisallee 21-23, D-04103 Leipzig, Germany
Ellagitannins are a large group of polyphenolic compounds widely distributed in plants. An increasing interest in the role of these particular metabolites in the therapeutic action of traditional medicines of China and Japan (1,2) has led to a rapid growth of knowledge in this area. Correlation of the biological activity of tannins with their structure has been made possible due to the isolation by reversed-phase HPLC methods and structural determination with high-resolution NMR and FAB-MS spectroscopic techniques.
In our preceding paper (3) we reported on the isolation and structure elucidation of ellagic acid 4-O-a -L-rhamnopyranoside, 6-O-galloyl-( a/ b)-O-glucopyrano se, 6-O-galloyl-2,3-(S)-hexahydroxydiphenoyl-( a/ b)-D-glucopyranose, corilagin, 3,3-di-O-methylellagic acid, ellagic acid, gallic acid, methyl gallate and 3-O-methyl-3,4-methylenedioxyellagic acid, isolated from the heartwood of Punica granatum L. (Punicaceae). Further chemical examination of polyphenolic metabolites of this plant has now resulted in the isolation of novel diellagic acid rhamnoside.
The 1,3-diellagic acid rhamnoside was isolated as yellowish white amorphous powder showing characteristic UV spectral data of an ellagic acid derivative. Complete acid hydrolysis resulted in ellagic acid and L-rhamnose. Its structure was established by ESI-MS and various 1d/2d ¹H/¹³C NMR spectral experiments.
To our knowledge, 1,3-diellagic acid rhamnoside is new and not yet described before.
1. E.Haslam, T.H.Lilley,Y.Cai, R.Martin and D.Magnolato, Planta Med., 1989,55,1.
2. T.Okuda, T.Yoshida and T.Hatano, J.Nat.Prod., 1989,52,1.
3. S.A.A.El-Toumy,M.S.Marzouk and H.W.Rauwald, Pharmazie, 2001 (in press)
Pharmacognosy and Chemistry of Medicinal Plants Department, National Research Centre, Dokki, (12311), Cairo, Egypt.
Acacia saligna Wendl is an ornamental plant belonging to the family Leguminosae. The compounds isolated from the genus Acacia are claimed to have many biological activities(1-3) In this work the chemical composition of the essential oil isolated by hydrodistillation from the leaves and flowers of Acacia saligna Wendl were investigated for the first time by means of gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS). A total of 35 and 29 components were identified from the leaf and flower oil, respectively. Qualitative and quantitative differences in the oil compositions were observed. The major constituents of the leaf oil were mainly the hydrocarbons nonadecane (18.81%), pentadecane (11.84%), heptadecene (11.58%) and heptadecane (11.054%) together with 14- methyl- pentadecanoic acid methyl ester (10.06%), whereas the flower oil was characterized by high percentages of oxygenated compounds except for a-phellandrene (8.94%) which is a hydrocarbon. The major components of the flower oil were trans-coniferyl alcohol (23.45%), g-6-methyl-ionone (14.62%), and 3-hexanol (7.36%).
Investigation of the hexane extract of the aerial parts of A. saligna resulted in the isolation and identification of 13 hydrocarbons, five fatty alcohols, the aldehyde octadecanal, as well as two pentacyclic triterpenes, namely methyl oleanonate and erythrodiol diacetate(4). a-Spinasterol, an antiinflamatory sterol isolated before from A. ehrenbergiana(5) , was also isolated from hexane extract of A saligna.
1. Zawahry, M.R. and Tawil, G.S., The Gazette of the Egyptian Paediaric Association, 175, (1956).
2. Ayoub, S.M. H., Fitoterapia, 53, 175, (1982).
3. Ayoub, S.M. H; Yankov, L.K. Fitoterapia, 56, 277-9 (1985).
4. Budzikiewicz, H.; Wilson, J.M. and Djerassi, C., J. Am. Chem. Soc., 85, 3688-3699, (1963).
5. Ahmed, F. A.Ph.D. Thesis, Faculty of Science, Chemistry Dept., Ain- Shams University, (2000).
!Institut für Entwicklungs- und Molekularbiologie der Pflanzen
§§Institut für Pharmazeutische Biologie
Heinrich-Heine-Universität Düsseldorf, Universitätsstrasse 1, D-40225 Düsseldorf, Germany
§Permanent address: Department of Pharmacognosy, Faculty of Pharmacy, Shiraz University of Medical Sciences and Health Services, Shiraz, I. R. Iran
There is great interest in lignans and their synthetic derivatives due to application in cancer chemotherapy and various other pharmacological effects1. In a screening for lignans we detected lignan-type components in extracts from callus, cell suspensions, roots and hairy roots of L. austriacum by HPLC due to their UV spectra. Separation by preparative HPLC from extracts of the untransformed roots resulted in the isolation of compounds 1 and 2.
1 was identified by GC-EIMS and NMR (1 H, 13C, COSY, HMQC and HMBC) as the arylnaphthalene lignanolide Justicidin B known from Justicia (Acanthaceae)2,3 and Haplophyllum (Rutaceae)4 species. 2 showed a very similar UV spectrum as 1 but appeared in the HPLC chromatogram at a higher retention time. GC-EIMS analysis revealed 2 as an isomer of 1 ([M]+ at m/z 364). 2 was finally determined to be the 5-demethoxy-3-methoxy isomer of justicidin B, i.e. 3,4-dimethoxy-3,4-methylenedioxy-2,7-cycloligna-7,7-dieno-9,9-lactone 5. The name samsamin is proposed for this compound, which, to the best of our knowledge, has not been described before. Justicidin B was not yet found in Linaceae.
(1) Ward, R. S. Nat. Prod. Rep. 1999, 16, 75-96.
(2) Abullaev, N.D.; et al. Chem. Nat. Compd. 1987, 23 , 63-74.
(3) Okigawa, M.; Maeda, T.; Kawano, N. Tetrahedron. 1970, 26, 4301-4305.
(4) Nukul, G.S.; et al. J. Nat. Prod. 1987, 50, 748-750.
(5) For numbering & nomenclature see Moss, G.P. Pure Appl. Chem . 2000, 72, 1493-1523.
Acknowledgements: We thank the Ministry of Health and Medical Education, I. R. IRAN, for a grant to A. Mohagheghzadeh., to I. Mehregan for identification and seed collection of L. austriacum as well as financial support by EC-project BIO-4-CT98-0451 "LIGNOCANCER".
*Department of Pharmacognosy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
**Department of Pharmacognosy, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran
Valeriana sisymbriifolia is one of the seven species of the Valeriana genus widely distributed in northern and central regions of Iran (1). It has been used therapeutically for more than ten centuries. The Valepotriates and terpenoids of the essential oil of this genus have been partly associated with der sedative, hypnotic and antispasmodic activities(2). A number of valepotriates have been previously isolated from the roots of some Valeriana species. In this paper, we report the isolation and structural determination of the dichlormethane extract of Valeriana sisymbriifolia using various chromatographic and spectroscopic methods and report the identification of two valepotriates.
From the dichloromethane extract we isolated a new valepotriate, 1- a-aceisovaltrate, and a new valepotriatehydrine, acetoxydeisovaleroxy-1-a -acetoxyisovaleroxy isovaltratehydrine together with known compounds, such as valtrate. Structural assignments of the compounds were based on spectroscopic methods (UV, IR, MS, H-NMR and C-NMR).
1-Rechinger KH.: Flora Iranica. NO 62. Akademische Druck-u. Verlagsanstalt, Graz. 1-23(1969).
2-Wichtl, M.: Teedrogen. 2. Auflage, Wissenschaftliche Verlagsgesellschaft mbH, Stuttgart, 79-82(1989).
*Departamento de Farmacología, Facultad de Farmacia, U.C.M. Madrid, Spain
*Centro de Biología Molecular Severo Ochoa, CSIC, Madrid, Spain
Extracts of a number of Bolivian species have been widely used in folk medicine for their biological properties. Among them, Satureja boliviana, known as Khoa , has been the focus of our research as source of new drug of antiviral interest (Abad et al, Gen Pharmacol, 32, 499, 1999) An ethanol-soluble fraction (SBS) and an ethanol-precipitate fraction (SBP-1) were preparated from the stems and leaves of this plant by hot water extraction and ethanol precipitation. According to a bioassay-guided fractionation using vesicular stomatitis virus (VSV) as the target model, the crude polysaccharide fraction SBP-1 exhibited antiviral activity. Confluent HeLa cells were infected with VSV at multiplicities of infection and incubated fo 24 h at 37 ºC (5% CO2). Just after addition of the virus, the fractions were added, and assayed at concentrations ranging from 25-500 ìg/ml. The antiviral activity was evaluated by determining the cytopathic effect (CPE) and measuring protein synthesis by means of [35S]-methionine incorporation. Dextrane sulphate was used as the reference compound at concentrations ranging from 0 to 200 µg/ml (60% inhibition at 200 µg/ml). SBP-1 was the most active fraction exhibiting antiviral activity against VSV at concentrations ranging from 5 to 125 ìg/ml (100 % at 250 ìg/ml). This fraction was further fractionated by the centavlon precipitation method (2) yielding three polysaccharide fractions termed SBP-2 (strong acidic), SBP-3 (weakly acidic) and SBP-4 (neutral). Enzymatic hydrolysis and sugar analysis led to the detection of the following sugars: galactose, arabinose, xylose and a sugar not yet identified.
Acknowledgement: Supported by DGICYT PB94-0148 and PR182-6738/96 UCM
Pharmacognosy and Chemistry of Medicinal Plants Department, National Research Centre, Dokki, Cairo, Egypt
Erucaria microcarpa Boiss . (Cruciferae) is a wild plant appears periodically in Egypt according to very hard weather conditions. Some volatile isothiocyanates were isolated from this herb(1), isopropyl isothiocyanate, butyl isothiocyanate and benzyl isothiocyanate in addition to the non-volatile 8-methyl sulphinyl octyl isothiocyanate (hirsutin). These compounds were tested for their antimicrobial activities. The total herb of E. microcarpa , when investigated for phenolic compounds, present in ethanolic extracts, revealed the presence of furopinnarin, glycyrine and 2,3-dihydroxydihydrosuberosin which was previously isolated from Coleonema album (2). These coumarins were identified by their spectroscopic properties in MS. 1H NMR and UV.
After defatting with petroleum ether, the dry herb was extracted with 70% alcohol and the general method for isolation and purification of flavonoids(3) was followed. In this way four compounds were isolated and identified as quercetin-3-O-galactoside, lucenin-l, robinetin and isorhamnetin-3-O-galactoside.
1. Hashem, FA. and Saleh, MM. (1999) Phytotherapy Research, 13: 329-332.
2. David, D., Michael, PV. and Cohan, P. (1972) Phytochemistry, 11: 705-7 13.
3. Mabry, TJ. Markham, KR. and Thomas, MB. (1970) The Systematic Identification of Flavonoids. Springer, New York pp. 53-97.
4. Budzikiewicz, H. Wilson, JM. and Carl Djerassi, (1963) J. Am Chemical Soc. Vol. 85: 3688-3699.
1 Department of Pharmaceutical Sciences, University of Antwerp, Belgium
2 Faculty of Pharmacy, University of Kinshasa, Democratic Republic of Congo
In the Democratic Republic of Congo some aromatic plants, such as Eucalyptus, Afromomum , Cymbopogon, Monodor and Ocimum species are used in traditional medicine to treat various diseases such as cough, upper respiratory tract infections and skin diseases [1]. No report about the antibacterial or antifungal activity of these plants growing in this African country was found in the literature. Only the insecticidal activity of Eucalytus saligna was described [2]. Thus, essential oils of 15 selected aromatic plant species were investigated for their chemical composition and for their putative antifungal activity.
Essential oils were obtained by hydrodistillation using adult fresh plant materials (leaves or seeds) collected in Kinshasa between February and December 1989. The chemical composition was studied by GC and GC/MS and the antifungal activity was evaluated by the diffusion method.
Results from the chemical study indicate that constituents such as a-pinene, b-pinene, limonene, 1-8-cineole, camphene, a-terpineol, g-terpinene and globulol were prevalent in difference concentrations. Some essential oils (5 µl per disc) exhibited an antifungal activity at different levels by producing a zone diameter of inhibition from 10 to 17, 11 to 18 and 13 to 19 mm against Candida albicans, Candida tropicalis and Aspergillus niger, respectively, after 24 h of incubation at 37 oC. A high antifungal activity was found in the essential oils of Eucalyptus alba leaves (14-17 mm) and Eucalyptus terticornis leaves (15-21 mm), followed by the essential oil of Eucalyptus camadulensis leaves, Eucalyptus globulus leaves and Monodora myristica seeds (10-15 mm) against both yeasts and fungi. The essential oil of Cymbopogon citratus leaves was inactive against C. albicans, but active against C. tropicalis and A. niger (12-15 mm). Ocimum americanum, Ocimum gratissimum and Afromomum stipulatum essential oils showed also a good activity (10-13 mm) against both selected yeasts and fungi. No correlation between the amount of major constituents such as 1,8-cineol, a -pinene or thymol and the antifungal activity was observed.
1. Kambu, K. Eléments de Phytothérapie Comparée. Plantes Médicinales Africaines. CRP-Kinshasa, 1990.
2. Kambu, K., Di Phanzu, N., Coune, C., Wauters, J.N., Angenot, L., Pl. Méd. Phytothér. 16, 34 38 (1982).
Department of Pharmaceutical Sciences, University of Antwerp, 2610 Antwerp, Belgium
Morinda morindoides (Baker) Milne-Redhead (Rubiaceae) (syn. Gaertnera morindoides ), commonly called Nkongabululu, Kongobololo or Nkama mesu, is one of the most popular medicinal plants used in the Democratic Republic of Congo for the treatment of various diseases such as malaria and amoebiasis, and as an antirheumatic agent. In relation to the traditional use against rheumatic pains, a bioassay-guided fractionation of both the ethyl acetate and n -butanol fractions obtained by partition from the crude 80% methanolic extract of the leaves, has already resulted in the isolation and structure elucidation of a series of flavonoids active as inhibitors of complement activation, radical scavengers and inhibitors of xanthine oxidase [1-4].
In the present communication the isolation and structure elucidation, using one- and two-dimensional NMR methods and mass spectrometry, of seven new iridoid lactones (1 - 7) from the same fractions is reported. These compounds are structurally related to the Plumeria iridoids, a relatively rare group of iridoids containing a spiro-lactone ring, which are found mainly in the Apocynaceae. In the Rubiaceae only one iridoid with this typical functionality has been identified, i.e. oruwacin from Morinda lucida.
|
|
R1 R 2 R3 R4 1 Me OH H glc. 2 Me OH H 6-acetyl-glc. 3 H OH H glu. 4 Me OH OMe glc. 5 Me =O H glc. 6 Me =O OMe glc. 7 Me OH H glc. 6,7- b-epoxide |
Cimanga et al., Phytochemistry 38, 1301 1303 (1995)
Cimanga et al., J. Nat. Prod. 58, 372 378 (1995)
Cimanga et al., Planta Med. 63, 220 223 (1997)
Cimanga et al., Pharm. Pharmacol. Commun. 5 , 419 424 (1999)
Department of Applied BioSciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland
The leaves of Warionia saharae Benth. & Coss. (Asteraceae), an endemic shrub from Morocco and South Oran in Algeria, are used in Morocco for inflammatory and gastrointestinal disorders [1]. An earlier phytochemical study on W. saharae focused on the main components of the essential oil and led to the isolation of eudesmol, linalool and nerolidol [2].
Using cytotoxicity against the KB cancer cell line (HeLa cells, ATCC CCL17) as a lead, bioactivity-guided fractionation of the MeOH soluble part of the Dichlormethane extract of W. saharae leaves led to the isolation of five new cytotoxic guaianolide type sesquiterpene lactones (1-5 ).
The structures of these guaianolides were deduced from 1D and 2D NMR spectroscopy ( 1H, 13C, DQF-COSY, HSQC, HMBC, ROESY), as well as mass spectrometry (EI- and HR-MALDI).
Bellkhdar, J., La pharmacopée marocaine traditionnelle, Médecine arabe ancienne et savoirs populaires, IBIS Press, 1997; 208
Ramaut, J. L., Hofimger, M., Dimbi, R. Corvisier M., & Lewalle, J. Main constituents of the essential oil of Warionia saharae Benth. & Coss.; Chromatographia 1985; 20: 193
Division of Pharmacognosy, Department of Pharmacy, P.O.B. 56, FIN-00014 University of Helsinki, Finland
Nine different Mentha species or hybrids cultivated at Maatalouden tutkimuskeskus, Mikkeli, Finland, were investigated for their essential oil composition. They constitute part of a mint collection established there in 1998. The species were: Mentha haplocalyx, M. crispa, M. arvensis var. piperascens, M. x verticillata, M. x aquatica, M. x dalmatica, M. sp. Frantsila , M. sp. Native Wilmet and M. sp. (Moroccan mint). The plants were grown in the field and leaves were harvested from August 15th to September 4th 2000.
There were great variations both in oil content (0.58-2.10% v/w) and oil composition between the different species. M. sp. Frantsila and Native Wilmet resembled peppermint containing menthol and menthone as main component, respectively. The higher menthone content in Native Wilmet could partly be explained by the fact that it was harvested 3 weeks later than Frantsila and the menthol content is known to increase during the vegetation period. Mentha crispa and M. x dalmatica had carvone as main component, 25 and 63%, respectively. M. arvensis var. piperascens contained menthol (54%) as main component as expected. The rest of the oil had unusual main components; M. x aquatica menthofuran (36%), M. x verticillata linalool (31%) and M. haplocalyx piperitone oxide (48%).
School of Pharmacy, Tokyo University of Pharmacy & Life Science, Hachioji, Tokyo 192-0392, Japan
The fruit of Prunus mume SIEB. et ZUCC. (Rosaceae) has been traditionally used as a medicinal food in Japan. In the course of our research on the pharmacologically active constituents of the fruits of P. mume we found the hypertensive actions of benzyl b-D-glucopyranoside (1) and chlorogenic acid (2), and we reported effects of 1 and 2 on plasma angiotensin converting enzyme and aldosterone levels in rats (1). Recently, we investigated the diuretic action of 1 and 2 in relation to their hypertensive action.
The constituents 1 and 2 were isolated from the ethanolic extracts of the fresh fruits of P. mume by the procedure reported before (2). After adminstration of NaCl solution or CMC suspension of 1 or 2 (0, 10, 30, 70 mg/kg) to mice (male ddY strain), the amount of urine produced was weighed at intervals of 20 min for 120 min. Both 1 (10 mg/kg) and 2 (70 mg/kg) showed strong diuretic effects making mice to excrete twice as much urine as compared mice. Furthermore, in the case of the administration of 1 together with spironolactone or potassium canrenoate, kown as antialdosterone uretics, remarkable combination effects were found. Consequently, these results suggest that the uretic action of 1 works as an aldosterone inhibitory substance.
1. Ina, H., Yamada, K., Miyazaki, T., XVth EFMC International Symposium on Medicinal Chemistry, Edinburgh, Scotland, 1998, Abstracts P. 102.
2. Ina, H., Yamada, K., Miyazaki, T., Natural Medicines 53, 109 (1999).
Cultivation and Production of Medicinal and Aromatic Plants Dept. & Pharmacognosy and Chemistry of Medicinal Plants Dept National Research Centre Dokki, (12311), Giza, Egypt
Solenstemma arghel (Dellile) Hayne is a glabrous shrub 0.4 1 m, much branched from the base with fleshy flowers, belonging to the family Asclepiadaceae. The plant is growing wild in the Egyptian desert east of the Nile and Sinai (1-3). In this work plant material was collected from shrubs growing on Gebel Elba region approximately 1200 km south of Cairo. The chemical composition of the essential oil isolated by hydrodistillation from S. arghel (4) leaves has been investigated by GC and GC MS(5 6). The essential oil of S. arghel, obtained in 0.5 0.9 % w/w on a dry weight basis, consisted approximately of 39.2 % linalool. Other oxygenated monoterpene compounds were identified, the principal members were a terpineol and trans geraniol which accounted for approximately 6.30 and 5.43 % of the oil, respectively. Additional oil constituents found in concentrations above 1 % were hexadecanal, elemol, 2-pentadecanone, 6, 10, 14 trimetyl cadinene, b eudesmol and hexadecanoic acid.
1. Tackholm, V. (1974). Student Flora of Egypt, Second edition. Cairo University
2. Boulos, L. (1995) Flora of Egypt. Checklist Hadara Publishing Cairo, Egypt
3. Boulos, L. (2000) Flora of Egypt VII Al Hadara Publishing Cairo, Egypt
4. Guenther, E. (196 1). "The Essential Oils", VIII. Robert. E D. Von Nostrand comp.,Inc. New York
5. Eyeele Mve Mba, C., Menut, C., Lamaty, G., Amvam Zollo, P. H. Techoumbougnang, F. And Bessilere (1994). Aromatic Plants of Tropical Central Africa. Part XIX. Volatile Components from Leaves of Two Lamiaceae from Cameroon: Leucas deflexa Hook and Solenstemon monostachyus (P.Beauv.) Briq Flavour and Fragrance Journal Vol. 9. 315 317
6. Adams, R. P.(1989). Identification of Essential oils by Ion Trap Mass Spectroscopy, Academic Press, New York
1Pharmacognosy Dept.
2Medicinal and Aromatic Plants Dept.
3Virology Dept.
National Research Centre, Dokki, Cairo,Egypt.
Taxodium distichum or bald cypress (Taxodaceae) is a large deciduous or ever green tree. Its light green feathery foliage and the narrow pyramidal habit which retain in cultivation give it very distinct appearance. Botanically it is closely related to the member of the Taxaceae family, including yew tree known to contain taxol as an antitumour agent. In meantime the seeds were reported to possess antitumour activities (1). The present work deals with its volatile oil and their antiviral activity.
Volatile oil was extracted from leaves by hydrodistillation using a Clevenger apparatus. Then the oil was analysed by GC/MS. Antiviral activity was tested against Herpes simplex virus type I using the method of Hundson (2) and virucidal activity was determined according to Silva et al (3).
Leaves of Taxodium distichum contain 0.52% volatile oil consists of 49 identified components which constitute about 99% and terpenene-4-ol (37.34%) was the major compound.
The percentage of virucidal activity using the plaque reduction assay was 73.68% corresponding to volatile oil. Terpenene-4-ol was found to be the compound responsible for the antiviral activity as reported by Bishop (4) on Melaleuca species, which confirms the result obtained in the present investigation. In addition, El-Tantawy et al. (5) reported the occurrence of 87% a-pinene in volatile oil of Taxodium fruit which had anti-inflammatory and antispasmodic activity .
Kupchan, S.M., Karim, S.M., and Marks, C.(1990).Tokyo Gakugei Kiyo 36:49-52
Hudson, J.B. (1990). CRC press, Inc., Boca Raton
Silva, O., Barbose, S., Diniz,A.,Valdeira, M. and Gomes, E. (1997) Inter. J. Pharm.35(1): 12-16
Bishop, C.D.(1 995). J. Essential Oil Res.,7:641-644
El-Tantawy, M.E., EI-Sakhawy, F.S., EI-Sohly, M.A. and Ross, S.A. (l999) J.Essentia1 Oil Res. 1(3): 641-644
Natural Products Research, Dr. Willmar Schwabe GmbH & Co, 76227 Karlsruhe, Germany
The roots of Pelargonium sidoides / reniforme have been utilized in traditional medicine of South Africa for the therapy of tuberculosis [1,2]. In modern phytotherapy Umckaloabo, an extract preparation of P. sidoides / reniforme , is used against infections of the respiratory tract and ear, nose, throat infections [3]. Because of their strong UV fluorescence at 366 nm coumarin derivatives are the most prominent compounds in extracts of Pelargonium species detectable by TLC [4]. In the butanol fraction of a 60% ethanol extract from roots of P. sidoides we found a weakly UV366 fluorescent spot, which apparently is not a coumarin. GC on silica yielded a compound which changed its Rf-value after boiling for crystallisation from ethyl acetate / acetone. Further GC on Sephadex LH-20 resulted in a pure compound which gave crystals from water with m.p. 202-3 °C.
|
|
1 R = H 2 R = CH2C6 H5 |
The 1H-NMR spectrum of 1 in DMSO-d6 shows doublets at 6.20 and 7.97 ppm with a coupling constant of 9.7 Hz being characteristic for 3,4-unsubstituted coumarins. Two methoxy and two phenolic hydroxy resonances correspond to 5,6,7,8-tetrasubstitution. The resulting molecular formula of C11 H10O6 was confirmed by elemental analysis. Comparison of the NMR data in acetone-d6 and of the melting points indicates, that 1 is different from 6,8-dihydroxy-5,7-dimethoxy-2H-1-benzopyran-2-one described by Kayser and Kolodziej [4]. The substitution pattern of 1 was clarified by NOESY spectroscopy of the dibenzyl ether 2 in DMSO-d 6. NOE correlation between H-4 and 5-OCH3, 5-OCH3 and 6-OCH3, 6-OCH3 and 7-OCH2 and, finally, 7-OCH2 and 8-OCH2 indicate doubtlessly the proposed structure 2. Work on the structure of the native compound is in progress.
1. Bladt, S. (1974) Dissertation, Univ. München
2. Bladt, S. (1977) Dtsch. Apoth.-Ztg. 117: 1655
3. Haidvogl, M., Schuster, R., and Heger, M. (1996) Z. Phytother. 17: 300
4. Kayser, O., and Kolodziej, H. (1995) Phytochemistry 39: 118
1Department of Pharmacognosy, School of Pharmacy, University of Athens, Panepistimio polis, Zografou, GR-157 71, Athens, Greece.
2Papanikolaou Research Center of Oncology. Department of Biochemistry, St. Savvas Hospital, GR-11 522, Athens, Greece.
Continuing our research on the chemical constituents of Centaurea sp., we here report on the isolation and identification of secondary metabolites from C. zuccariniana DC, belonging to the section Acrolophus (Cass.) DC [1].
Their isolation was proceeded according to the Bohlmann isolation method [2]. The residue was prefractionated by VLC (Coll & Bowden, 1986) on silica gel, using cyclohexane-EtOAc-Me 2CO mixtures of increasing polarity as eluents to give several fractions. Further purification on RP-HPLC (MeOH-H2O 1:1) yielded compound 1.
Compound 1 is a new naturally occurring eudesmanolide. Its structure was elucidated by spectroscopic methods, particularly high-field NMR spectroscopy, as 8 a-hydroxy-sonchucarpolide. Besides compound 1, three known sesquiterpene lactones were isolated, namely elemanolide 8 a-(3,4-dihydroxy-2-methylene-butanoyloxy)-dehydromelitensine ( 2 ), as well as the germacranolides cnicin (3) and 4-acetylcnicin ( 4), major constituents of the plant.
Cytotoxic/cytostatic activity of the isolated compounds was tested against various human cell lines (SF268, H460, MCF7, OVCAR3, BLD1) [3]. In addition to the isolated compounds, we tested other sesquiterpene lactones isolated from other Greek endemic Centaurea sp. [4-5]. The germacranolides showed moderate to strong cytostatic activity which varies depending on the side chain. It is noteworthy that the elemanolide 5 showed high cytostatic and cytotoxic activity.
Georgiadis Th. (1980), Contribution à létude phylogénétique du genre Centaurea L. (Sectio Acrolophus (Cass.) DC.) en Grèce. Thesis, Université de Provence- Aix Marseille I, 286 pp.
Bohlmann F., Zdero C., King R. M. and Robinson H. (1984), Phytochemistry 23, 1979-88.
Maswadeh H., Hatziantoniou S., Demetzos C., Dimas K., Georgopoulos A., Rallis M. (2000) Anticancer Research 20:4385-4390.
Skaltsa H., Lazari D., Panagouleas C., Georgiadou E., Garcia B., Sokovic M. (2000), Phytochemistry 55, 903-8.
Skaltsa H., Lazari D., Garcia B., Pedro J., Sokovic M. and Constantinidis T. (2000), Z. Naturforsch. 55C, 534-9.
1 Department of Pharmacognosy, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11000 Belgrade, Yugoslavia
2 Department of Pharmacy, Division of Pharmacognosy, University of Athens, Panepistimioupolis Zografou, 15771 Athens, Greece
The genus Valeriana L. (Valerianaceae) is represented by 8 species in the flora of Serbia. The main compounds isolated from Valeriana species are ester iridoids (valepotriates), cyclopentanoid sesquiterpens (valerenic acid and its derivatives) and essential oil. The roots and rhizomes of V. officinalis are used for the preparation of phytomedicines that are employed as mild sedatives. Besides, they can be used as gastrointestinal sedatives, poison antidotes, and deodorants.
In the framework of our chemical and biological investigation on the volatile components of Yugoslavian plant species, we report the chemical composition of the essential oil obtained from roots and rhizomes of Valeriana officinalis L. s.l. from wild population and from the aerial and underground parts of V. montana . Voucher specimens have been deposited at the Herbarium of the Department of Pharmacognosy, Faculty of Pharmacy, University of Belgrade.
Plant material was collected during the flowering period in July 2000 on the Tara Mountain in the west part of Serbia. Essential oils were obtained by hydrodistillation. The chemical analysis of the oils obtained from the underground and aerial parts was carried out using GC/MS. Compounds were identified on the basis of their Kovats indices and mass spectra compared with those of authentic samples and/or NIST/NBS, Wiley libraries and literature data.
In the oil obtained from underground parts of V. officinalis (sample A) 53 components were identified representing 90.7 % of the total oil. The main constituents were: valerianol (57.3%) and bornyl acetate (11.3%). In the oils obtained from aerial (sample B) and underground parts (sample C) of V. montana the main constituents were: isovaleric acid (sample B-44.1%, sample C-27.8%), 3-methyl isovaleric acid (sample B-34.0%, sample C-41.3%) and myrtenol (sample B-5.3%, sample C-6.4%).
M. Josifovic. Flora of Serbia, Vol. V, Serbian Academy of Sciences and Arts, Belgrade (1974)
Ockedon, D. J.: Flora Europaea (Tutin, T.G., et al.), Vol. 4, University Press, Cambridge, p. 55 (1972).
Adams, R. P.: Identification of Essential Oil Components by Gas Chromatography/Mass Spectroscopy. Allured Publishing Co Illinois, USA (1995).
Bos, R.: Analytical and phytochemical studies on valerian and valerian based preparations, Thesis, State University of Groningen, The Netherlands (1997).
1Department of Pharmacognosy, School of Pharmacy, University of Athens, Panepistimiopolis,Zografou, GR-157 71, Athens, Greece
2 Department of Applied BioSciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich, Winterthurerstr. 190, 8057 Zurich, Switzerland
Continuing our research on the chemical constituents of Stachys sp., we report here the results of the investigation of Stachys ionica Halácsy, a greek endemic species, belonging to the subsection Swainsonianeae Bhattacharjee [1]. The subcosmopolitan genus Stachys L. contains more than 270 species and is justifiably considered as one of the largest genera of the Lamiaceae [2]. Greece is certainly an area particularly rich in taxa [3].
In addition to several known flavonoids, an elemane derivative (-)8 a-hydroxy-elemol ( 1) was isolated. So far, the (+)-enantiomer of the latter has been isolated [4-8]. The structure of the compounds was elucidated by spectroscopic methods, particularly highfield NMR spectroscopy (1H, 13C, COSY, HSQC-TOCSY, NOESY, HSQC and HMBC).
|
|
|
1 |
Persson, D., 1981. Biosystematics of Stachys swainsonii Benth. (Lamiaceae) and its relations to some other chasmophytic Stachys species. Ph.D. thesis, University of Lund, Lund.
Bhattacharjee, R., 1980. Taxonomic studies in Stachys : II. A new infrageneric classification of Stachys L. Notes from the Royal Botanic Garden Edinburgh 38, 65-96.
Greuter, W., Burdet, H. M., Long, G., 1986. Med-Checklist, Vol. 3. Editions des Conservatoire et Jardin botaniques de la Ville de Genève, Genève.
De Pascual, Teresa J., San Feliciano, S., Egido, T., Barrero, A F., 1977, Anales de Quimica 151.
Mata, R., Navarrete, A., Alvarez, L., Pereda-Miranda, R., Delgado, G., Romo de Vivar, A., 1987, Phytochemistry 26, 191.
Mossa, J. S., Muhammad, I., El-Feraly, F. S., Hufford, C. D., 1992, Phytochemistry 31, 2789.
Ohashi, H., Asai, T., Kawai, S., 1994, Holzforschung 48, 193.
San Feliciano, A., Medarde, M., Lopez, J. L., Miguel del Corral, J. M., Puebla, P., Barrero, A. F., 1988, Phytochemistry 27, 2241.
1Semmelweis University, Department of Pharmacognosy
2Technical and Economical University of Budapest, Department of Chemical Engineering, Budapest, Hungary
Supercritical fluid extraction is a technique which is now widely applied to prepare extracts of vegetable origin. Traditional methods for extracting natural terpenoids from medicinal plants generally use large volumes of organic solvents. The process is slow, the solvent must then be separated and evaporated to give the residual extract. In this process may degrade some sample constituents. In view of increasing environmental concerns about the use of liquid solvents in the extraction of natural products, there has been growing interest in alternative extraction techniques using supercritical fluids.
The aim of this work was to examine the effect of supercritical fluid extraction on some pharmacologically significant medicinal plants containing non volatile terpenoids such as the antiinflammatory triterpenoids, proteine- kinase C inhibitor sesquiterpene- g-lactones and phytosterols. We also wished to show possibilities for the production of extracts in the most beneficial composition.
The effect of sample preparation, extraction conditions (pressure, temperature, time) and separation circumstances (pressure, temperature) was examined on the yields and composition of bioactive constituents.
The role of pressure and termperature was studied in the 32 full factorial design. Model plants were: Taraxacum officinale Wiggers et Weber, Cnicus benedictus L., Chrysanthemum parthenium L. (Beruh), Vitex agnus castus L., Origanum vulgare L., Thymus serpyllum L. and Thymus vulgaris L.
The products gained by supercritical fluid extraction were different from the traditional ones concerning their appearance, content and composition of bioactive constituents therefore they have to be considered as special raw materials of plant origin. Using optimal pressure and temperature as parameters the sesquiterpene- g-lactone parthenolide (Chrysanthemum parthenium), the triterpenoide taraxasterole and b-amyrin (Taraxacum officinale), as well as the phytosterols (Taraxacum officinale, Thymus serpyllum, Oreganum vulgare) were quantitatively extracted and selectively enriched in supercritical extracts. We could produce extracts containg 16.0-24.5 % b-amyrin from dandelion, 9.1 % triterpenes from chaste tree, 1.1-2.0 % parthenolide from feverfew and products rich in phytosterols from chaste tree (0.1-0.5 %), thyme (0.2-0.2 %), dandelion (1.5 %). The sesquiterpene-g-lactone cnicin was absent in supercritical extracts prepared by carbon dioxide. When 4 % ethanol was added to the C02 flow to the column, it eluted the lactone fraction containing cnicin (Cnicus benedictus). The yields of these bioactive constituents in the supercritical extracts were several times higher than those corresponding extracts wich were prepared by traditional methods. To characterise the extractability of the individual compound an index was indroduced (log P* 100/M.W.)
Acknowledgement: The work was supported by FKFP-0169, ETK T-08-158/99, OTKA T-0300344.
Kéry Á., Rónyai E., Simándi B., Lemberkovics É., Keve T., Deák A., Kemény S.: Chromatographia 49, 9/10, 503-508 (1999)
Simándi B., Oszagyán M., Lemberkovics É., Kéry Á., Kaszás J., Thyrion F., Mátyás T.: Food Research Int. 31, 10, 723-728 (1999)
Kristó T.Sz., Terdy P., Simándi B., Kéry Á.: Olaj, szappan, kozmetika 49, 93-98 (2000)
Oszagyán M., Simándi B., Kéry Á., Lemberkovics É.: Olaj, szappan, kozmetika 49, 98-101 (2000)
1 Faculty of Pharmacy, Mahidol University, Sri-Ayudhya Rd., Bangkok 10400, Thailand
2 Faculty of Sciences, Mahidol University, Phra RamVI Rd., Bangkok 10400, Thailand
3 Laboratory of Pharmacognosy and Plant Chemistry, Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka 814-01, Japan
4 Pharmazeutisches Institut der Universitaet Freiburg, Albertstrasse 25, D-79104 Freiburg, Germany
The pressed juice from Murdannia loriformis herb has been used for 20 years by patients who are suffering from various types of cancers, especially breast and uterus cancers, in Thailand.
The whole plant, 30 kg, was collected, dried and ground. The powdered drug, 1.8 kg, was successively extracted with petroleum ether, chloroform and ethanol, respectively. The ethanol extract was chromatographed on Diaion HP20 (MCI gel CHP 20 P) column. The methanol fraction from the column yielded glycosides, which were further purified chromatographically. A phytosteryl glucoside (G1a) and a glycosphingolipid (G1b) were isolated. The structures were identified as 3-b - O-D-glycopyranosyl-24x -ethyl-cholest-5-ene (G1a) and 1-b -O-D-glycopyranosyl-2(2¢ -hydroxy-6¢ -ene-cosamide)-sphingosine (G1b). G1a was cytotoxically inactive, using the MTT cytotoxicity test, whereas G1b showed a moderate cytotoxicity with ED 50 of 16 m g/ml against human breast and colon cancer cell lines, and also immunomodulatory properties. An HPLC method to quantify G1b was established. The method was applied to determine the G1b concentration in the pressed juice.
University of Joensuu, Department of Biology, Natural Product Research Laboratory, P.O.Box 111, 80101 Joensuu, Finland
The seedling stage of plants is very vulnerable to environmental and biological constrains. In birch, phenolic compounds are possible means in defence. The concentrations of phenolic compounds are affected by internal factors, such as plant age, and environmental conditions, such as fertility of the habitat. We examined phenolic compounds in the first true leaves of 2.5-week-old seedlings of mountain birch ( Betula pendula ssp. czerepanovii) grown in three different nitrogen levels.
Seeds from an individual mountain birch in Botanical Gardens of Joensuu were germinated and seedlings grown in perlite:vermiculite (1:1) mixture. Seedlings were fertilised with 3, 30 and 140 ppm of nitrogen in addition to other nutrients according to Ingestad 1962. Dry weights of individual plant parts were measured and phenolics were determined from methanol-extracted samples by HPLC.
The growth of seedlings was significantly increased by increased nitrogen. The concentrations of condensed tannins and total flavonol-glycosides were significantly lower at the highest nitrogen level. Individual flavonol-glycosides had different reactions from highly significant to no difference in concentration in different nitrogen levels. The fertilisation effect on hydrolysable tannins was variable, and there was no clear effect on phenolic acids. The results indicate that secondary compound levels of seedlings can be affected by nutrients even at the very beginning of ontogeny. The effects are mainly in accordance with the carbon/nutrient balance hypothesis. The distinctly decreased carbon allocation to secondary compounds (condensed tannins) in nitrogen-rich growing conditions, however, seemed to be in no clear proportion to the relatively small increase in growth.
Ingestad, T. 1962. Macro element nutrition of pine, spruce, and birch seedlings in nutrient solutions. Meddelanden från Statens Skogsforskningsinstitut 51:1-131.
1Department of Zoology and Marine Biology, University of Dar-es Salaam, P.O Box 35064, Tanzania
2Freie Universität Berlin, Institut fur Pharmazie, Abt. Pharmazeutische Biotechnologie, Kelchstraße 31,12169 Berlin, Germany
3Schaper & Brümmer GmbH & Co KG, Analytische Entwicklung, Bahnhofstraße 35, D-38225, Salzgitter Ringelheim, Germany
4Freie Universität Berlin, Department of International Animal Health, Luisenstr. 56, D. 10117 Berlin, Germany
5Faculty of Veterinary Medicine, Sokoine University of Agriculture, P.O. Box 3021 Morogoro, Tanzania
The phytochemical profile of Aloe secundiflora (Aloeaceae) leaf exudates has been examined leading to the identification of 8 major compounds with two main constituents. A. secundiflora is used ethnoveterinary in some parts of Kenya and Tanzania. Analytical HPLC- MS studies of the exudate have revealed that it comprises of a mixture of phenolic compounds mainly anthraquinones and chromones together with a low content of polysaccharides or aliphatic compounds. The results showed that A. secundiflora contains significant amounts of aloenin, aloenin B, isobarbaloin, barbaloin, aloinoside A and B and other not yet identified aloesin derivatives. Main peaks represented 80% of the area under the curve (AUC) in HPLC. The main compounds were aloenin B and the aloins A and B, followed by barbaloin and isoaleosin D.
Seven fractions were obtained by quantitative HPLC, covering the major peaks from prior qualitative HPLC analysis. All fractions were introduced in antibacterial testing, exhibiting antibacterial activity to various degrees. The aloenin containing fraction showed highest activity against Salmonella gallinarum and other microorganisms at a MIC of 5 mg/mL. To continue bioguided screening, the aloenin containing fraction was purified by quantitative HPLC yielding three subfractions. In one aloenin was enriched up to 35 % (m/m), a second subfraction consisted of 50% (m/m) of a mixture of aloesin and aloin derivatives. Both were tested, and the aloenin containing subfraction showed the highest antibacterial activity.
The aim of the study was to verify the ethnoveterinary importance of the exudate of this Aloe species as used locally in the prophylaxis of Newcastle disease virus and in control of other bacterial infections in chickens including fowl typhoid. The high content of aloenin and aloin that have high antimicrobial activity in the exudate gives a first line of evidence for the use of A. secundiflora in ethnoveterinary practice.
aFaculty of Pharmacy, Philadelphia University, Amman, Jordan
bInstitut für Pharmazeutische Biologie der Heinrich-Heine-Universität, Düsseldorf, Germany
cSwiss Tropical Institute, Basel, Switzerland
Human African sleeping sickness caused by the protozoan parasite Trypanosoma brucei rhodesiense in East Africa and by T. b. gambiense in West and Central Africa infects 300 000 persons annually.1 While resistance of this pathogen to the currently available synthetic trypanocidal agents represents an increasing problem, many plants used for the treatment of Trypanosomiasis in African traditional medicine have not been studied systematically although they may contain agents with possibly novel mechanism of action.
The decoction of the leaves of Argemone mexicana L. (Papaveraceae) is widely used in Sudanese traditional medicine for the treatment of trypanosomiasis. In vitro evaluation of the chloroform, ethyl acetate and n-butanol extracts exhibited no trypanocidal activity while the aqueous extract displayed the most prominent antitrypanosomal activity against T. b. rhodesiense (IC 50 = 0.09µg/ml). Bioactivity directed fractionation of this extract resulted in the isolation of the quaternary benzo[c ]phenanthridine alkaloid, sanguinarine 1 (IC50=0.05 µg/ml).
|
|
|
|
1 |
2 |
Our earlier investigation 2 of the antiplasmodial activity of the leaves A. mexicana exhibited similar patterns. This activity was entirely confined to the aqueous extract (IC50 = 4.5 µg/ml) when tested against chloroquine resistant strains of Plasmodium falciparum. Sanguinarine 1, in one of our previous studies, exhibited some antiplasmodial activity against chloroquine resistant strains, which however was very weak as compared to its anti-trypanosomal activity (IC50 = 2.65 µg/ml).
In vitro cytotoxicity for sanguinarine 1 on three cell lines (HT-29, MRC-5 and NBZa) was lower than anti-trypanosomal activity by factors of 60, 17 and 70, respectively (IC50 values: 3.0, 0.85 and 3.5 µg/ml). 1 thus appears to show selective toxicity towards T. b. rhodesiense .
The structurally related quaternary alkaloid, berberine 2, also a major constituent of A. mexicana, has been previously subjected to in vitro anti-trypanosomal evaluation (IC50=0.4 µg/ml) using the same protozoal strain 3. The much lower activity of this protoberberine alkaloid in comparison with sanguinarine, coupled with the bioactivity directed fractionation approach we used in the present study, indicates that sanguinarine 1 is the main alkaloid responsible for the antitrypanosomal activity of A. mexicana . Further work to study the mechanism of action involved in the trypanocidal activity of sanguinarine and other related benzophenanthridine alkaloids is underway.
WHO, 1998. Control and surveillance of African trypanosomiasis. WHO Technical Report Series No. 881.
Noureldin, A. et al., 2001 J. Ethnopharmacology (submitted).
Freiburghaus, F. et al., 1996 J. Ethnopharmacology 55, 1-11.
1 Division of Upland Crops, National Yeongnam Agricultural Experiment Station, R.D.A. Milyang 627-130, Korea
2 Korea Rearch Institute of Bioscience and Biotechnology, P.O. Box 115, Yusong, Daejon 305-600, Korea
3 Korea Research Institute of Chemical Technology, P.O. Box 107, Yusng, Daejon 305-343, Korea
Two
new steroidal saponins, along with two known spirostanol saponins
were isolated from the methanolic extract of the rhizomes of
Asparagus oligoclonos . The structures of the new steroidal
saponins were elucidated based on spectroscopic analysis, including
1H, 13C- NMR and two dimentional NMR
techniques. New steroidal saponins were characterized as (25S)-5
b-spirostan-3 b,17
a-diol-3-O-
b-D-glucopyranosyl-(1->2)-
b-D-glucopyranoside and (25S)-5b
-spirostan-3 b,17
a-diol-3-O-
b-D-xylopyranosyl-(1->2)-[
a-L-rhamnopyranosyl-
(1->4)]-b-D-glucopyranoside
which appeared to contain a new type of aglycon. The two known
spirostanol saponins were (25S)-5b-spirostan-3
b-ol-3-O-b-D-glucopyranosyl-(1->2)-
b-D-glucopyranoside and (25S)-5
b -spirostan-3b-ol-3-O-
b-D-glucopyranosyl-(1->2)-[b
-D-xylopyranosyl-
(1->4)]-b-D-glucopyranoside.
Two new saponins were evaluated for cytotoxic activity on the five
human tumor cell lines.
Department of Phytochemistry, Institute of Pharmacology, Polish Academy of Sciences, Pl-31-343 Kraków, Poland
In the course of our chemical investigation of Cichorium intybus L. (Asteraceae) we have isolated a total of 11 guaiane-type sesquiterpene lactones from root and leaf extracts. In addition, the roots of the plant have yielded one germacranolide and one eudesmanolide. The 1H NMR spectrum of the eudesmanolide revealed almost identical chemical shift values of the key protons to those previously reported for magnolialide (1) [1], cichoriolide A (5) [2] and cichopumilide (9) [3].
This unexpected similarity of the 1H NMR spectral data of the above mentioned sesquiterpene lactones necessitated more detailed structural analysis of the isolated eudesmanolide. The compound was eventually proved to be magnolialide (1) by means of spectral methods, including 2D NMR experiments (NOESY, COSY, HETCOR), and by comparison of optical rotation values. The carbon resonances of 1 were indistinguishable from those of 5, after reassignments of the signals from C-1, C-2 and C-3 according to the structure of 1 , giving evidence in favour of the identity of 1 and 5. Additional support for the above, as well as for the identity of 1 and 9 was obtained from a comparison of physical and spectral data reported for 11b, 13-dihydroderivatives 2, 6 and 10. Again, the chemical shift values of 13C NMR signals of 2 were identical to those of 6 when reassigned in accord with the structure of 2. The 1H NMR spectral properties of the three dihydroderivatives were also very similar, in particular when more complete data of 10 were compared with those of 2. Accordingly, compounds 5, 6 and 9, 10 have been reassigned the revised structures 1 and 2, respectively. This, in turn, give rise to the revision of cichorioside A (7) and sonchuside C ( 8) structures, now correctly represented by 3 and 4, respectively. Compounds 5, 7 and 8 were reported from C. intybus and/or C. endivia [2], while compounds 9 and 10 were found in C. pumilum ( a subsp. of C. endivia) [3].
|
|
|
|
|
1 R = H, X = CH2 2 R = H, X = H, a Me 3 R = Glc, X = CH2 4 R = Glc, X = H, a Me |
5 R = H, X = CH2 6 R = H, X = H, a Me 7 R = Glc, X = CH2 8 R = Glc, X = H, a Me |
9 X = CH2 10 X = H, a Me |
El-Feraly, F. S., Chan, Y. M., Benigni, D. A. (1979) Phytochemistry 18: 881-882
Seto, M., Miyase, T., Umehara, K., Ueno, A., Hirano, Y., Otani, N. (1988) Chem. Pharm. Bull. 36: 2423-2429
El-Masry, S., Ghazy, N. M., Zdero, Ch., Bohlmann, F. (1984) Phytochemistry 23: 183-185
1Frohme-Apotheke, Frohmestraße 14, D-22457 Hamburg, Germany
2Övelgönner Straße 7, D-20257 Hamburg, Germany
The European Pharmacopoeia includes about 100 monographs on herbs and herbal preparations. Quality control of the raw material is divided into an identification and a quantification section. The quantification is covered by HPLC methods, and TLC methods are mostly restricted to fingerprint identifications.
In this paper we present the TLC identification of the roots of Devils claw (Harpagophytum procumbens, Pedaliaceae) which are used in traditional medicine of Southwest Africa. The lead substance is the iridoid compound harpagoside. Another marker substance is 8-para-coumarylharpagide (8-pCHG). A discrimination of the two species H. procumbens and H. zeiyheri is based on the 8-pCHG-identification number, which means the relation of harpagoside to 8-pCHG.
By optimizing the chromatographic system proposed in the monograph, we achieved separations which even serve for purposes of quantifying the lead substances. In addition to this we tested the biological activity directly on the plate, e.g. the inhibition of the cholinesterase.
The inhibitory effect of the active substances is determined by the suppression of the enzymatic hydrolysis of 1-naphthyl acetate to 1-naphthol and acetic acid. In consequence the complementary coupling reaction of 1-naphthol with Fast Blue B salt resulting in a blue diazonium salt is inhibited, too. Thus the active substances appear as white zones on the plate. This was found to be the case in the extract of freshly harvested roots of H. procumbens.
1 Department of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, TR-06100 Ankara, Turkey
2 Department of Medicinal Chemistry, College of Pharmacy, University of Utah, Salt Lake City, Utah 84112, USA
In the flora of Turkey, the genus Globularia (Globulariaceae) is represented by eight species [1]. G. alypum is used as diuretic, laxative, stomachic and tonic [2], and G. trichosantha for the treatment of hemorrhoids in Anatolian folk medicine [3]. In our previous papers, we described the isolation of phenylethanoid glycosides, several iridoid and bisiridoid glycosides from G. trichosantha [4,5], and iridoid glycosides, phenylethanoid glycosides and sucrose esters from G. orientalis [6]. In the continuation of chemical studies on Turkish Globularia species, we have now investigated an endemic species, G. davisiana O. Schwarz. Chromatographic studies on the aerial parts of G. davisiana resulted in the isolation of a new iridoid, davisioside (1), in addition to eight known iridoid glycosides, asperuloside (2), alpinoside (3), geniposide (4), globularin (5), globularicisin (6), 10-O-benzoylcatalpol (7 ), lytanthosalin (8) and melampyroside (9). The structures of 1-9 were established by spectroscopic methods. Davisioside ( 1 ) represents a new iridoid structure lacking the double bond between C-3 and C-4. This type of iridoid aglycone is rarely found in nature.
1. Davis, P.H. (1982) Flora of Turkey and East Aegean Islands; University Press: Edinburgh, Vol. 7: pp.27-31.
2. Baytop, T. (1984) Therapy with Medicinal Plants (Past and Present), Istanbul University Publications: Istanbul, No. 3255, pp 419.
3. Sezik, E., Tabata, M., Yesilada, E., Honda, G., Goto, K., Ikeshiro, Y. (1991) J. Ethnopharm. 35: 191-196.
4. Çalis, I., Kirmizibekmez, H., Rüegger, H., Sticher, O. (1999) J. Nat. Prod. 62: 1165-1168.
5. Çalis, I., Kirmizibekmez, H., Sticher, O. (2001) J. Nat. Prod. 64: 60-64.
6. Kirmizibekmez, H., Sticher, O., Tasdemir, D., Ireland, C., Çalis, I. (2000) International Congress and 48th Annual Meeting of the Society for Medicinal Plant Research, Sept. 3-7, 2000, Zurich, Switzerland. P2A/49.
1Department of Applied BioSciences, Institute of Pharmaceutical Sciences, ETH Zurich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
2ZELLER AG, Seeblickstrasse 4, CH-8590 Romanshom, Switzerland
The roots of Valeriana officinalis belong to the most often used components of herbal medicinal products with indication for sleep disorders [1]. Despite of many compounds tested, it is yet unclear which groups of components are responsible for the sedative effect of this plant. Actually, the valerenic acids play an important role as leading substances in the quality control of medicinal products.
Valepotriates are another group of characteristic substances in Valeriana [2]. In the seventies they were supposed to contribute to the effective principle [3]. Later, their cytotoxic and mutagenic potential was discovered and discussed [2, 4, 5], so that they are not desirable anymore in medicinal products of Valeriana officinalis. Due to their instability valepotriates are usually absent in fluid extracts [2].
A TLC method was developed for the simultaneous qualitative proof of valepotriates and valerenic acids. With this method drug and extract can be easily controlled and compared, respectively, regarding their content of valepotriates. In the usual case of absence of valepotriates in the extract, its identity will be confirmed by the proof of valerenic acids. The TLC method was validated by a HPLC method of Bos [2] with analogous results for the presence of valepotriates and valerenic acids in the measured drug and extracts.
A screening for valepotriates of different swiss valerian extracts will be presented.
1. Schilcher H, Kammerer S (2000) Leitfaden Phytotherapie. Urban & Fischer Verlag München, p 290
2. Bos R (1997) Analytical and phytochemical studies on valerian and valerian based preparations. Thesis, Rijksuniversiteit Groningen, Netherlands
3. Thies PW (1966) Zur Konstitution der Isovalerensäureester Valepotriat, Acetoxyvalepotriat und Dihydrovalepotriat. Tetrahedron Lett 11, 1163-1170
4. Bounthanh C, Bergmann C, Beck .JP, Haag-Berrurier M, Anton R (1981) Valepotriates, a new class of cytotoxic and antitumor agents. Planta Med 41, 21-28
5. Meier B (1992) Baldrian - Traditionelles Schlafmittel. Der Allgemeinarzt 12, 1102-1112
1Department of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, TR-06100 Ankara, Turkey
2Department of Medicinal Chemistry, College of Pharmacy, University of Utah, Salt Lake City, Utah 84112, USA
Our previous phytochemical studies on Turkish Digitalis species (Scrophulariaceae) afforded a variety of glycosides [1,2]. From D. cariensis Boiss. ex Jaub. & Spach, we reported several phenylethanoid glycosides [3]. Further investigations of the aerial parts of this plant resulted in the isolation of a furostanol glycoside (1) and a pregnane glycoside (2). On the basis of spectral methods (NMR, MS), compound 1 was identified as 3-O-(-lycotetraosyl)-26-O-(-D-glucopyranosyl)-(25R )-22-methoxy-5-furostane-3,26-diol. Compound 1 is identical to uttroside A, whose structure was previously established by chemical methods and IR spectroscopy [4]. This represents the first report of uttroside A (1 ) from the genus Digitalis. We also report the full NMR assignments of uttroside A (1), which are lacking in the literature. Compound 2 was identified as digifologenin-3-O--D-glucopyranosyl-(14)--D-oleandropyranoside. 2 is a new natural product and cariensisoside is proposed as the trivial name. Cariensisoside (2) has very similar NMR data to those of glucodigifolein [5], except it contains -D-oleandropyranose instead of -D-diginopyranose in the carbohydrate moiety.
1. Çalis, I., Tasdemir, D., Sticher, O., Nishibe, S. (1999) Chem. Pharm. Bull. 47: 1305-1307.
2. Çalis, I., Akbay, P., Kuruüzüm, A., Yalçin, F.N., Sahin, P., Pauli, G. (1999) Pharmazie 54: 926-930.
3. Kirmizibekmez, H., Ersöz, T., Çalis, I. (2000) XIIIth International Symposium on Plant Originated Crude Drugs, Sept. 20-22, 2000, Istanbul, Turkey. P-55.
4. Sharma, S.C., Chand, R., Sati, O.P., Sharma, A.K. (1983) Phytochemistry, 22 (5): 1241-1244.
5. Liedtke, S., Wichtl, M. (1997) Pharmazie 52: 79-80.
1Bundesanstalt für Züchtungsforschung an Kulturpflanzen, Institut für Qualitätsanalytik, Neuer Weg 22/23, 06484 Quedlinburg, Germany
2Universität Kassel, Fachgebiet Agrarbiodiversität, Steinstraße 11, 37213 Witzenhausen, Germany
3DRAGOCO, Gerberding & Co.AG, Dragocostraße, 37601 Holzminden, Germany
4Universität Leipzig, Institut für organische Chemie, Johannisallee 29, 04103 Leipzig, Germany
At present, characterisation of fennel distinguishes chemotypes by the presence or absence of trans-anethole and estragole [1] or by the quantitative graduation of these phenyl propenyl derivates and fenchone [2]. This classification also includes one type of Foeniculum vulgare spp. piperitum in which piperitenone oxide was detected as main component for the first time [3].
The forms of Foeniculum vulgare spp. piperitum now discovered contain methoxy substituted estragoles as main components. In the first case g-asarone was found at 72 % in the essential oil. The essential oil of the other form contains 15 % methyl eugenol and 41 % g-asarone. The origin of both types is Portugal.
The seeds of F. vulgare ssp. piperitum type 1 and type 2 contain 3.1 % and 2.2 % essential oil, respectively. Methyl eugenol was identified by comparison with the reference compound g-asarone by GC/MS, H1- and C13-NMR.
1. H. Krüger and K. Hammer: Chemotypes of Fennel ( Foeniculum vulgare Mill.), J. Essent. Oil Res., 11, 79-82 (1999).
2. J. Bernáth, É. Németh, A. Kattaa and . Héthelyi: Morphological and Chemical Evaluation of Fennel (Foeniculum vulgare Mill.) Populations of Different Origin. J. Essent. Oil Res., 8, 247-253 (1996).
3. A. Badoc, G. Deffieux, A. Lamarti, G. Bourgeois and J.-P. Carde: Essential Oil of Foeniculum vulgare Mill. (Fennel) subsp. piperitum (Ucria) Cout. Fruit. J. Essent. Oil Res., 6, 333-336 (1994).
1 Laboratory for Pharmacognosy, Phytochemistry and Toxicology, Dept. of Pharmaceutical Sciences, Vrije Universiteit Brussel (VUB), Laarbeeklaan 103, B-1090 Brussels, Belgium
2 Laboratory for Microbiology, Academisch Ziekenhuis-Vrije Universiteit Brussel (AZ-VUB), Laarbeeklaan 101, B-1090 Brussels, Belgium
3 Department of Pharmaceutical Sciences, University of Antwerp (UIA), Universiteitsplein 1, B-2610 Antwerp, Belgium
Epinetrum villosum (Excell) Troupin (Menispermaceae) is a twinning liana found in secondary forests of the Democratic Republic of Congo (DRC). In Lomela area (DRC), a root bark decoction of this plant is orally administered for the treatment of diarrhoea and dysentery [1]. The crushed leaves are used as a wound dressing for fast healing [2]. The plant is also traditionally used for the treatment of pains and mental strain [3].
Crude root bark extracted with solvents of increasing polarity (diethyl ether, 70% methanol and water) were tested against ten diarrhoeal bacteria using the microtiter plate dilution method [4]. The methanol extract was found to be active against Campylobacter jejuni and Campylobacter coli having a MIC value of 62.5 µg/ml. The water extract exhibited a moderate activity against Shigella sonnei , Shigella dysenteriae, Campylobacter jejuni and Campylobacter coli with MIC values of 1000, 1000, 500 and 1000 µg/ml respectively [5].
The methanol extract was fractionated by column chromatography on silica gel 60 (Merck) using a CHCl 3-MeOH gradient. An alkaloid (cycleanine) belonging to the bisbenzylisoquinoline family has been recently isolated in our laboratory and its structure has been elucidated by means of TLC, UV and NMR spectroscopy.
Cycleanine
Further antibacterial testing of this alkaloid will reveal if cycleanine is responsible for the activity shown by the MeOH crude extract.
1. Longanga Otshudi A., Vercruysse A., Foriers A. (2000) Journal of Ethnopharmacology. 71: 411-423.
2. Troupin, G (1951) Flore du Congo Belge et du Ruanda-Urundi. 2, p220.
3. Parvez M., Abdur Raman., Salma Rahman and Ogbeide O.N. (1994) Journal of Chemical Society of Pakistan. 16(4): 257-260.
4. Mitscher LA., Leu, R.P., Bathela M.S., Wu W-K. (1972) Lloydia. 35:157-176.
5. Longanga Otshudi A., Foriers A., Vercruysse A., Van Zeebroek A., Lauwers S. (1999) Phytomedicine. 7(2):167-172.
1Chemistry of Natural Products Department, National Research Centre, Dokki, Cairo, Egypt
2 School of Pharmaceutical Sciences, University of Shizuoka 422-852, Shizuoka, Japan
3 Pharmacognosy Department, Faculty of Pharmacy, Helwan University, Cairo, Egypt
The pharmacological importance associated with saponins has encouraged us to examine their natural occurrence in the Araliaceae member Oreopanax guatemalensis. Our pharmacological investigation, on the saponin-containing fraction from leaves and stems of O. guatemalensis showed that it exhibited significant analgesic, antipyretic and anti-inflammatory activities. Our phytochemical investigation has resulted in the isolation of seven new saponins named Oreopanaxsaponins I-VII (4,6,5,7,1,3,2). The saponins were isolated and purified by CC and HPLC techniques. The identification of the saponin structures in general was allowed by the use of positive FAB mass spectrum and ID and 2D NMR techniques. HOHAHA difference, 1H-1H COSY, NOE difference, HMQC and HMBC spectra established the structure of the aglycone and the monosaccharide units. Their identification were further confirmed by total acid hydrolysis followed by HPLC and GLC analyses. NOE difference measurements and HMBC spectrum determined the interglycosidic linkages and the attachment of the sugar chain to the aglycone. 1H and 13C NMR signal assignments were made possible by the combined use of 1H-1H COSY, HOHAHA difference, HMQC and HMBC experiments.
R1 R2 R3 R4 R1 R2
1
OH H
5
2
OH H
6
3
OH H
4 Ara H OH
R1 R2
7
Ara = a-L-arabinopyranosyl; Ara(f) =a -L-arabinofuranosyl; Rha = a-L-rhamnopyranosyl; Glc = b-D-glucopyranosyl; Xyl = b-D-xylopyranosyl.
1Department of Pharmacognosy, K. Marcinkowski University of Medical Sciences, 10 Sieroca Str., 61-771 Poznan, Poland
2Institute of Physics, Department of Technical Physics, Poznafi University of Technology, 13A Nieszawska, 60-995 Poznan, Poland
The genus Viburnum L. (Caprifoliaceae) comprises about 120 species. Some of them are cultivated for decorative purposes, whereas some are used in medicine. Viburnum prunifolium L. and Viburnum opulus L. barks are considered to be uterine spasmolytic and supposed to calm and alleviate uterine pain in menstrual disorders (dysmenorrhoea and amenorrhaea). In folk medicine the drugs are also employed in treating menopausal complaints. To our knowledge only few phytochemical studies on Viburnum lantana L. have been reported in the literature (1). Therefore, our interest focused on the methanolic extracts obtained from the aerial parts of this species.
From the leaf extract we isolated the following compounds: luteolin, apigenin, chrysoeriol 7-O-b-glucosides; apigenin 7-O- a-rhamnoside (1®6) glucoside and also amentoflavone (biflavone) which were all identified by different techniques (total and partial hydrolysis as well as spectral methods: UV, 1 H, 13C NMR). The flower extract afforded only flavonol derivatives. Among them were: kaempferol and quercetin 3-O-b -galactosides; 3-O-b-glucosides; 3-O- a-rhamnoside (1®6) b-glucosides; 3-O-a-rhamnoside (1>2) b-glucosides.
Since amentoflavone exhibits interesting biological activities we decided to study its molecular interactions and spectral properties in artificial membranes (2, 3). The sample was embedded in a PVA (polyvinyl alcohol) film not only for orientation, but also in order to minimize oxidation and aggregation problems (polymer content was 15%). The results obtained for amentoflavone in DMF were compared with those from samples in the film. These studies are still in progress.
Acknowledgement: Part of our work was supported by the project (PP) DS-62-176/2001
1. Godeau R.P., Pelissier Y., Sors G. & Fouraste I. (1978). Plantes med. Phytother. 12, 296.
2. Bialek-Bylka G.E., Brown J.S. & Manikowski H. (1987). Photosynthetica 21, 182-184.
3. Siodmiak J. & Frackowiak D. (1972). Photochem. Photobiol. 16, 173-182.
1 Department of Pharmacognosy, Faculty of Pharmacy, Al-Azhar University, Assiut 71524, Egypt
2 Department of Pharmacognosy, Faculty of Pharmacy, Assiut University, Assiut 71526, Egypt
A new triterpene 2b, 3b , 16a, 28-tetrahydroxy-olean-12-ene-23-oic acid 2 was isolated from Gladiolus segetum Ker-Gawl bulbs (Iridaceae) with medicagenic acid 1 and their structures were established by different spectroscopic methods.
The air-dried powdered bulbs were extracted with methanol. The concentrated extract was fractionated with n-hexane and chloroform. The mother liquor was passed through Diaion HP-20 and fractionated with distilled water and methanol to yield a methanolic saponin fraction which was subsequently hydrolyzed. The aglycones were extracted with EtOAc and the extract then evaporated. The yellowish residue was chromatographed on a silica gel column. Fractions eluted with n-hexane-EtOAc (1:1) were rechromatographed on Sephadex LH-20 to yield 1 and 2 .
The 1H-NMR spectrum of 2 showed six methyl singlets and an olfenic proton. The 13C-NMR signals indicated that 2 is D12 oleanene derivative, DEPT measurements showed six CH 3, nine CH2 comprising an oxygenated one, eight CH including three oxygenated methines and seven quaternary carbons, of which one is a carboxyl. This carboxyl group was located at C23 due to identical resonances of C4 and C24 compared with 1. The CH2OH group could be located at C28 due to downfield shift of C17. Comparison of NMR data of 1 and 2 revealed the existance 2, 3 dihydroxy system in both two compounds. However 2 showed an extra secondary hydroxy function which was located at C16 based on absence of normal resonance of C16 and downfield shift of C15 compared with 1. Compound 1 was identified as medicagenic acid (mp, IR, 1H and 13C-NMR).
|
|
R1 |
R2 |
|
1 |
COOH |
H |
|
2 |
CH2OH |
OH |
Chemistry of Tanning Materials and Proteins Department, National Research Centre, Dokki, Cairo, Egypt
In the continuation of our studies of the polyphenolic constituents present in Solenostemma arghel leaves [1,2], due to its medicinal importance [3], a new kaempferol glycoside (1); kaempferol-3-O-â -D-galactoside (1à2)-arabinosyl (1 à2)(1à5) di-rhamnosyl; was isolated together with four known kaempferol glycosides namely: - kaempferol-3-O- â-D-galactoside(1à2)- á-L-rhamnosyl; 3-O- â-D-galactoside; 7-O- â-D-glucoside and 4`-O- â-D-glucoside. All the above compounds were isolated and purified through chromatographic procedures and their chemical structures were identified by means of chemical analysis and spectroscopic methods (UV, 1 H and 13C-NMR)[4,5].
(1à2) rhamnosyl
R= galactoside(1à2)
arabinosyl
(1à5) rhamnosyl
Michael, H.N. (1998). Asian Journal of Chemistry, 10 (4), 1038-1040.
Michael, H.N., Shafek, R.E., Ishak, M.S. (2000). Polyphenols Communications 2000, XXth International Conference on Polyphenols, Freising-Weihenstephan (Germany), September 11-15, 2000.
Boulos, L. Medicinal Plants of North Africa, Reference Publications, Inc., Michigan. U.S.A. (1983).
Markham, K.R., Techniques of Flavonoid Identification. Academic Press, London (1982).
Harborne, J.B., Mabry, and T.J. The Flavonoids Advances in Research. Chapman and Hall, London (1982).
1Department of Pharmacognosy
2Department of Physical Chemistry, Faculty of Pharmacy
The Medical University of Warsaw, Banacha 1, 02-097 Warsaw, Poland
Convallaria majalis L. (Liliaceae) is widely distributed in the forests of Europe. The overground parts of C. majalis contain cardenolidic glycosides and preparations thereof are used in treatment of heart insufficiency. In the roots and rhizomes several steroidal saponins were found. Tschetsche et al. [1] isolated convallamaroside, of which the aglycone is convallamarogenin, identified as D25-5 b,20b,22 a-spirosten-1b-diol. Besides convallamaroside, other steroidal spirostanol and furostanol saponins (13 compounds) were isolated from the roots and rhizomes of C. majalis by means of column chromatography [2]. New steroidal spirostan/spirosten sapogenins have been isolated recently with 1a,2b ,3b,5b -tetraol substitution pattern [3].
As a part of our studies on C. majalis we here report the isolation and structural characterisation of the saponin.
The powdered roots and rhizomes of C. majalis, (1900 g) were macerated for 48 h with 50% methanol. The extract was obtained was partitioned between CHCl 3 and n-BuOH with H2O. 800 mg of compound A (mp. 258-262 oC) was obtained by column chromatography from the faction of the less polar compounds.
The structure of this compound was elucidated by analysis of 1H and 13 C NMR spectral data, including 2D techniques (HETCOR, HMBC, ROESY) and 13C CPMAS NMR. Compound A was found to be D25-5b -spirosten-1a,2 b,3b ,5b-tetraol 3-O- b-D-galactopyranoside.
Tschesche R., Schwartz H. and Snatzke G., Chem. Ber. (1961) 94,1699
Nartowska J. and Strzelecka H., Acta Polon.Pharm. (1983) 25, 650
Nartowska J., Wawer I., in preparation
National Research Centre, Department of Phytochemistry, Cairo, Egypt
Extracts of some Euphorbia species have been used as folk medicine for treating diarrhea and wounds [1]. Investigations of the tannins of these species have revealed the wide distribution of geraniin, a dehydroellagitannin together with many of its dimeric derivatives, including euphorbins A - F [2]. These dimers were first isolated from E. humifusa whose extract was found to contain also ellagic acid 4-O-ß-glucopyranoside. These results have prompted us to investigate the aqueous ethanolic extract of Euphorbia consobrina N. E. Br. cultivated in Egypt for its polyphenolics.
1: Ellagic acid 4-O-b-xylopyranoside: R = H
2 : Ellagic acid 3,3-dimethyl ether 4-O-b -xylopyranoside; R= OCH3
The present study describes the isolation and structure elucidation of the new 4-O- bxylopyranosides of ellagic acid (1) and its 3.3-dimethyl ether (2). Together with these two new compounds, the known polyphenolics ellagic acid 3-O-b- glucopyranoside, corilagin, geraniin, 2.6-di-O-galloyl- b-glucospyranose, gallic acid, ellagic acid, 3,3-dimethoxy ellagic acid, p-methoxycoumaric acid, the 3-O-a -rhamnopyranoside of both kaempferol and quercetin in addition to the 3-O- b-glucopyranoside of the latter have been also isolated and characterized. The structures of all compounds have been established by conventional methods of chemical and spectral analysis and confirmed by ESI-MS 1D-1 H,13C-NMR and 2D-COSY experiments. The 13C-NMR data of the isolated ellagic acid glucoside was recorded and assigned here for the first time.
1. I. Agata, T. Hatano, Y. Nakaya, T. Sugaya, S. Hishibe, T. Yoshida and T. Okuda, Chem. Pharm. Bull., 39(4), 881 - 883 (1991).
2. T. Yoshida, Y. Amakura, Y. Liu and T. Okuda, Chem. Pharm. Bull., 42(9), 1803 - 1807 (1994).
1 Department of Pharmacognosy, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt
2 Current address: Pharmacia Corp, 700 Chesterfield Parkway North. Chesterfield MO, 63198, USA
3 Institute of Chemistry, Neuchâtel University, Neuchâtel, Switzerland
The aerial parts of Cleome droserifolia are reputed for their use in folk medicine as a hypoglycemic drug(1); in addition they are used topically for the treatment of wounds and dermatitis(2). Its hypoglycemic activity was recently confirmed by a clinical study(3).
In this study, we have isolated three diterpenoid acid derivatives of the dolabellane skeleton. The three compounds were isolated after extraction and extensive chromatography of the light petroleum and acetone extracts of the plant. The chemical structure of the isolated compounds was determined mainly by spectral as well as chemical methods.
The three compounds (I, II & III) are esters of 3-hydroxy-3-methyl glutaric acid. Compound I & II are acid derivatives, while compound III is the methyl ester of compound I.
It should be noted that compound III is a novel natural product, and this is the first report of this class of compounds in the genus Cleome and the second report of its presence in higher plants(4).
1 Yang, S.S.; Mabry, T.J.; El-fishawy, A.M.; El-Kashoury, E.A.; Abd ElKawy, M.A.; Soliman, F.M., (1990) Egypt. J. Pharm. Sci. , 31, 443
2 Batanouny, K.H. (1999) Wild medicinal plants in Egypt, first edition, Palm Press
3 Ghanem, M. M. (1999) M.S. thesis, Faculty of Medicine, Alexandria University. The short term effect of use of some medicinal plants used in folk medicine on non-insulin-dependent diabetes mellitus (NIDDM) patients (type II diabetes)
4 Mohamed, K.M.; Ohtani, K.; Kasai, R. and Yamasaki, K. (1995) Phytochemistry, 39, 151
Acknowledgement: We would like to thank the Swiss government for providing the grant for Hoda M. Fathy during her study in Switzerland during 1998/1999.
* Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey
** Department of Biology, Faculty of Education, Gazi University, 06500 Ankara, Turkey
Chemical and pharmaceutical studies in the past three decades have revealed that aquatic organisms are significant sources of biologically active substances with a potential to be developed into new drugs and other useful products. Marine algae have also been confirmed to be rich in lipoids. On the other hand, monounsaturated and polyunsaturated fatty acids are well known to possess protective effects on human health.
The fatty acid content of several fresh-water (Cladophora fracta, C. glomerata, Zygnema pectinatum, Maugeotia sp., Potomogeton perfoliatus , Vaucheria sessilis, and Spyrogyra gratiana) and marine ( Padina vickersiae, Posidonia oceanica, Halopteris scoparia , Scinaia furcellata, Sargassum natans) algae growing in Turkish waters were examined by using GC-MS in order to evaluate their nutritious value. The results given in the following table will be discussed in this presentation.
Table. Fatty acid composition of the lipoid extracts of the marine/aquatic plant and alga species.
|
Fatty acid
|
Retention time (Rt, min) |
Relative percentages of the fatty acids in the marine/aquatic plant and alga species studied |
|||||||||||
|
HS* |
PV* |
SF* |
SN* |
PO* |
CF* |
CG* |
ZP* |
SG* |
VS* |
PP* |
M* |
||
|
Myristic |
8.43 |
- |
- |
- |
- |
- |
5.90 |
- |
- |
- |
6.11 |
- |
- |
|
Capric |
11.41 |
1.00 |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
- |
|
Palmitic |
11.77 |
7.49 |
5.61 |
9.00 |
7.48 |
6.37 |
4.38 |
7.54 |
31.05 |
11.23 |
13.57 |
6.28 |
3.69 |
|
Oleic |
13.30 |
5.06 |
6.74 |
2.80 |
18.31 |
4.89 |
19.84 |
30.73 |
53.44 |
16.97 |
- |
11.19 |
4.75 |
|
Stearic |
13.44 |
10.82 |
- |
4.02 |
- |
5.23 |
- |
8.84 |
- |
- |
2.25 |
- |
- |
*(HS= Halopteris scoparia, PV= Padina vickersiae, SF= Sciniaia furcellata, SN= Sargassum natans, PO= Posidonia oceanica, CF= Cladophora fracta, CG= Cladophora glomerata, ZP= Zygnema pectinatum, SG= Spirogyra gratiana, VS= Vaucheria sessilis, PP= Potomogeton perfoliatus, M= Maugeotia sp.)
Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330, Ankara, Turkey
Two polyunsaturated fatty acids, linoleic and a-linolenic acids, are required by human body. These fatty acids, called essential fatty acids (EFAs), are critical components of nerve cells, cell membranes, and chemical messengers known as prostoglandins. As EFAs cannot be produced endogenously, the body must obtain them through dietary sources. Nuts, known to be rich in fat, are widely consumed in Turkey as snack and are also exported.
Continuing our search on fatty acids of edible Turkish plants in order to find new sources for essential fatty acids, we have characterized fatty acid compositions of the nut oils of Corylus avelana (Betulaceae), Castanea sativa (Fagaceae), Persea gratissima (Lauraceae), Juglans regia (Juglandaceae), Pinus pinea (Pinaceae), and Arachidis hypogaea (Fabaceae). Detailed results will be given in this presentation.
Table. Fatty acid content of the fixed oils of the nuts analyzed.
|
Plants |
Fatty acids identified and their relative percentages in the nut oils analyzed |
|||
|
Palmitic acid (Rt= 11.47) |
Linoleic acid (Rt=12.96) |
Oleic acid (Rt=13.03) |
Stearic acid (Rt=13.10) |
|
|
Corylus avelana |
10.54 |
8.78 |
77.36 |
3.32 |
|
Juglans regia |
2.66 |
25.26 |
47.16 |
5.45 |
|
Persea gratissima |
5.13 |
4.87 |
52.59 |
- |
|
Pinus pinea |
- |
54.33 |
25.70 |
5.06 |
|
Castanea sativaa |
24.10 |
4.46 |
11.45 |
4.33 |
|
Castanea sativab |
14.79 |
7.80 |
29.55 |
5.11 |
|
Arachidis hypogaea |
11.60 |
75.15* |
-* |
3.93 |
a Kernel used
b Shell used
* Mixed with oleic acid
1Laboratoire de Photochimie Moléculaire et Macromoléculaire, UMR CNRS 6505, Chimie des Huiles Essentielles, Université Blaise Pascal de Clermont et ENSCCF, France
2Department of Pharmacognosy, Faculty of Pharmacy, University of Belgrade, Yugoslavia
The genus Tanacetum L. (tribe Anthemidae Cass., Asteraceae) consists of about 150 species, which predominantly distributed in Europe and in Mediterran. In the Flora of Serbia, the genus is represented with 6 species [1]. We investigated the essential oil of Tanacetum macrophyllum (Waldst. & Kit.) Schultz Bip. from the vicinity of the Vlasina Lake (Southern Serbia). Aerial parts of the plant were collected during the period of full flowering. Once harvested, the plant material was dried at room temperature.
The isolation of the essential oil was carried out by steam distillation according to Proceeding III of the Yugoslavian Pharmacopoeia IV [2]. We established that the essential oil yield was 0.24% (in g of essential oil per 100 g of dried plant material).
The composition of the obtained essential oil was elucidated by a combination of GC and GC-MS. Component identification was carried out by comparing the experimental retention indices with those of standard substances added to the mixture. The identification was also confirmed by GC-MS. In the essential oil 33 components were identified and determined (77.69% of the total amount). In the analyzed oil monoterpenes were dominating (71.35%). Sesquiterpenes were present in a considerably smaller quantity (4,82%). The principal compounds in the oil were oxygenated monoterpenes: cis-chrysanthenol (26.26%), 1,8-cineole (12.28%), borneol (7.57%), trans-chrysanthenyl acetate (5.16%), linalool (4.10%) and camphor (3.09%). The analyzed essential oil differs significantly from the oil of Tanacetum millefolium (L.) Tzvelev from Greece [3] and the oil of Tanacetum vulgare L. from Argentina [4].
1. Josifovic, M. (Ed.): The Flora of FR Serbia, Vol. VII. SANU, Belgrade, 1975.
2. Yugoslavian Pharmacopoeia, Pharmacopoea Jugoslavica editio quarta, Ph. Jug. IV, National Institute for Health Protection, Belgrade, 1984.
3. Souleles, Chr., Stamatakou, M. (1991) Planta Med. 57: 92.
4. Gallino, M. (1988) Planta Med. 54: 182.
1 Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich, CH-8057 Zürich, Switzerland
2 Institute of Pharmaceutical Chemistry and Pharmaceutical Technology, Karl-Franzens-University of Graz, A-8010 Graz, Austria
Abietic acid (I) is one of the most important resin acids in conifere species after technical processing of the oleoresin [1] . It plays an effective role in the wound healing of trees by hardening after evaporation of the essential oil, accompanied with antibacterial and antifungal activity.
Due to the extensive use of resin acids in several products of everyday life, e.g. paper sizing, printing ink, adhesives, glues, technical resins and even cosmetics, an increasing number of people suffer from contact dermatitis and asthma [2 ]. Several studies reveal, that not resin acids themselves, but mainly their oxidation products cause these allergenic reactions. The amazing reactivity of colophony under very moderate conditions is one reason for its rank within the top ten international documented allergens [3 ].
In this work oxidation reactions of resin acids under very simple conditions were investigated. The idea was to reconstruct oxidation conditions, which might appear in aqueous ethanolic solutions. Abietic acid (I), a weak allergen [4], was oxidized in ethanol/water-dilution in a closed flask by room temperature and daylight. After two years it gave the following three new oxidation products (II -IV) - structure elucidation was done by means of NMR and IR spectroscopy as well as EI mass spectrometry:
[1 ] Ullmanns Encyclopädie der technischen Chemie, 4th Edition, Volume 12, Verlag Chemie 1987, 529-533
[2] A. M. R. Downs, J. E. Samson, Contact Dermatitis 1999, 41, 305-310
[3] B. M. Hausen, J. Mohnert, Contact Dermatitis 1989, 20, 133,143
[4] S. Sadhra, I. S. Foulds, C. N. Gray, British Journal of Dermatology 1996, 134, 662-668
*Pharmacognosy and Chemistry of Medicinal Plants Department
**Medicinal and Aromatic Plants Department
National Research Centre,12311, Dokki, Cairo, Egypt
Thuja oriantalis (Cupressaceae) is a permanent ornamental tree used in tradional Korean and Chinese medicine to treat hypertension and related disorders (1). It has tonic, expectorant and antitussive effects and has been used to treat various diseases (2). It has been reported to have antitumour activity in man (3). Total phenolic flavonoids extracted from Thuja occidentalis showed mutagenic effects (4).
In the present study, purpurin has been isolated from the alcoholic extract (70%) of Thuja oreantalis leaves for the first time from this genus and the Cupressaceae in general. Quercetin, quercetin 3-glucoside and the phenolic acids (gallic and elagic) were also isolated. Moreover, stigmasterol, b-sitosterol as well as palmetic (14.3%), oleic (7.0%), and eicosanoic (37.6%) were present in the petroleum ether extract as determined by GLC.
The insecticidal activity of the alcoholic extract (70%) showed significant activity on the mortality rate of Callosobruchus maculatus (92.5%) and on female Tetranychus urtica (70%) while the petroleum ether extract showed 80% mortality on Spodoptera littoralis and 100% molluscicidal activity on Biompholaria alexandrina snails.
Purpurin
Fazal, M. and Husain, S.I. (1991) New Agriculturist, (2), 111-112
Tegtmeter, M. and Harnischfeger, G. (1994) Eur. J. Pharm. Biopharm. 4(5), 337-40
Kai, Y. and Suran, E.P. (1990) Mokuzai Gakushi J. Japan wood Research Society, 36(3), 218-224
Kamil, W.M. (1998). Journal of Biological Sciences Research 19(1) 65-70
1 Institute of Pharmacognosy, University of Vienna, Centre of Pharmacy, Althanstraße 14, A-1090 Vienna, Austria
2 Institute of Pharmaceutical Chemistry, University of Graz, Universitätsplatz 1, A-8010 Graz, Austria
Crataegus-derived drugs are commonly used in treatment of heart weakness. In the pharmacopeia of Europe for hawthorn leaf with flower as well as fruit C. monogyna JAQUIN emend. LINDMAN, C. laevigata (POIRET) DE CANDOLLE, C. pentagyna WALDSTEIN et KIT. ex WILLD., C. nigra WALDSTEIN et KIT., C. azarolus L., as well as the hybrids of C. monogyna and C. laevigata are mentioned. For the identification of hawthorn species and hybrids besides morphological studies the differences in chemical profile especially the flavonoid complex are investigated (1). Whereas the flavonoid pattern of C. monogyna and C. laevigata is already well characterised, the hybrids were not yet studied. For this reason the flavonoids of C. x macrocarpa HEGETSCHWEILER, a hybrid of C. laevigata and C. rhipidophylla GANDOGER were investigated.
Air dried powdered leaves and flowers of C. x macrocarpa were extracted with 40% methanol. The removal of accompanying apolar substances was achieved by dissolving the dried methanolic extract in water and extraction with petroleum ether. The aqueous solution was extracted successively with ethyl acetate and n-butanol to pre-fractionate the flavonoids due to their different polarity. The residued aqueous layer was evaporated and fractionated by CC on Sephadex LH-20®. Elution was performed with water-methanol-mixtures of increasing polarity. Further purification was achieved by preparative HPLC (Nucleosil RP-18, 5µ, 20 x 250mm) with water-methanol as well as CC on Sephadex LH-20® with methanol.
By those procedures hyperoside, rutin, luteolin-7-O-b-D-glucoside, vitexin and vitexin-2 -O-rhamnoside could be identified by comparison with authentic substances. Besides these flavonoids, which are already well known in C. monogyna and C. laevigata, two new flavonoids were isolated. Their structures, identified as eriodictyol-7-O- b-D-glucuronide and luteolin-7-O-b -D-glucuronide, were elucidated by UV, API-MS, H1-NMR and 2D-NMR-techniques as HSQC, H,H-COSY and HMBC. Glucuronic acid was additionally confirmed by enzymatic hydrolysis of eriodictyol-7-O-b -D-glucuronide and luteolin-7-O-b-D-glucuronide with b-glucuronidase (2). Eriodictyol and luteolin were identified by comparison with authentic substances.
Eriodictyol-7-O- b-D-glucuronide and luteolin-7-O-b -D-glucuronide are two significant markers of the flavonoid complex of C. x macrocarpa which may have chemotaxonomical importance for further studies; they are reported for the first time from Crataegus sp.
Kurzmann, M. (1999) PhD thesis, University of Erlangen, Germany
Pemp, E. (2000) Diploma thesis, University of Vienna, Austria
*Institute of Plant Nutrition and Soil Science, Federal Agricultural Research Centre (FAL), Bundesallee 50, D-38116 Braunschweig, Germany
** Dreluso Pharmazeutica, Dr. Elten & Sohn GmbH, Markt 6, D-31840 Hessisch-Oldendorf, Germany
Glucotropaeolin has been proven to be effective against bacterial infections. Therefore Tropaeolum majus (L.) is used among others as a herbal medicine against urinary tract infections. High and stable glucotropaeolin contents within the plants are desirable for improved economics of the production process. Drug concentration varies in different in plant parts and changes during the vegetation period as shown in table 1.
Table 1: Mean concentrations of glucotropaeolin in different plant parts of Tropaeolum majus (L.) at different growth stages (n=36)
Besides this the glucotropaeolin content can be influenced by applying appropriate agricultural practices (table 2).
Table 2: Influence of sulfur and nitrogen fertilization on glucotropaeolin content of Tropaeolum majus (L.) at the start of flowering
Department of Pharmacognosy, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran
The genus Stachys (Lamiaceae) is distributed in mild regions of the Mediterranean and South-West Asia. About three hundred Stachys species are reported; 34 of them are found in Iran of which 13 are endemic.
Several Stachys species are used in Iranian folk medicine as medicinal plants. The genus Stachys has been the subject of only a few studies about essential oils. As part of a program of chemical investigation on volatile oil of Iranian medicinal plants, the oil constituents of Stachys laxa Boiss. & Buhse collected from Mazandaran Province (in North of Tehran) is reported for the first time.
The volatile oil of the air-dried aerial parts of S. laxa, isolated by hydrodistillation, was analyzed by capillary GC and GC-MS. Identification of components of the oil were based on GC retention indices relative to n-alkanes and computer matching with the WILEY275.L library, as well as by comparison of the fragmentation patterns of the mass spectra with those reported in the literature and, whenever possible, by co-injections with authentic compounds.
The aerial parts of S. laxa yielded 0.1% of a yellowish oil. Thirty-three components were identified. Germacrene-D (40.1%), b-caryophyllene (16.7%), b-phellandrene (5.5%), caryophyllene oxide (4.6%), linalool (3.2%) and ?-cadinol (2.6%) were found to be the major constituents of the oil.
Previous studies (1-4) on volatile oils of members of the Stachys genus showed varying compositions. The oil of S. laxa is characterized by a high content of sesquiterpenes with germacrene-D as the major component.
A. Cakir, M. E. Duru, M. Harmandar, S. Izumi and T. Hirata, The volatile constituents of Stachys recta L. and Stachys balansae L. from Turkey. Flavour Fragr. J., 12, 215-218 (1997)
M. Harmandar, M. E. Duru, A. Cakir, T. Hirata, and S. Izumi, Volatile constituents of Stachys obliqua L. (Lamiacea) from Turkey. Flavour Fragr. J., 12, 211-213 (1997)
M. E. Duru, A. Cakir, M. Harmandar, S. Izumi and T. Hirata, The volatile constituents of Stachys athorecalyx C. Koch. from Turkey. Flavour Fragr. J., 14, 12-14 (1999)
J. C. Chalchat, S. D. Petrovic, Z A. Maksimovic and M. S. Gorunovic, Essential oil of the herb of Stachys recta L., Lamiaceae from Serbia. J. Essent. Oil Res., 12, 455-458 (2000).
Chemistry of Tanning Materials and Proteins Department, National Research Centre, Dokki, Cairo, Egypt
A large population of different genera is contained in the family Myrtaceae, whereby polyphenolic compounds and various flavonoid classes are widely distributed in these genera [1]. The leaves stem and flowers of P. guaijava show a considerable degree of antibacterial activity [2] as they yield antibacterial substances such as flavonol glycosides [3,4,5], ellagitannins [6] and novel gauvins [7].
To our best knowledge only scarce information was reported on the seeds of Psidium guaijava. Repetitive chromatography of the diethyl ether extract of its seeds afforded ten phenolic and flavonoid compounds including one new acylated flavonol glycoside, quercetin-3-O-b-D (2``-O-galloyl glucoside)-4`-O-vinylpropionate. The structures of the new compound together with the other known compounds namely; 8,3`-dimethoxy gossypetin, quercetin-3-O-glucoside, the aglycones kaempferol, quercetin and myricetin; methyl gallate, ellagic, gallic and caffeic acids, were elucidated through different chemical and physical methods as well as spectroscopic analysis (UV, 1H- and 13 C-NMR). Also, an investigation of the pharmacological activities of the extract was carried out whereby it showed high inhibitory effect in vitro EAC cells and a moderate one against P338 leukemia cells.
Harborne, J. B. The Flavonoids: Advances in Research , Chapman and Hall, London (1982)
Watt, J. M. and Breyer-Brandwijk, M. G. The Medicinal and Poisonous Plants of Southern and Eastern Africa, E. and S. Livingstone LTD. Edinburgh and London (1962).
El-Khadem., H.; Mohammed, Y. S. J. Chem. Soc., 3320-3 (1958)
Seshadri, T. R.; Vasishta, K. Phytochemistry, 4(6), 989-92 (1965)
Kandil, F. E.; El-Sayed, N. H.; Micheal, H. N.; Ishak, M. S.; Mabry, T. J., Asian J. of Chemistry, 9 (4), 871-872 (1997)
Okuda, T.; Yoshida, T.; Hatano, T.; Yazaki, K. and Ashida, M., Phytochemistry, 21(12), 2871-4 (1982)
Okuda, T.; Yoshida, T.; Hatano, T.; Yazaki, K.; Ikegami, Y.; Shingu, T., Chem. Pharm. Bull., 35(1), 443-6 (1987).
Department of Pharmaceutical Biology, Institute of Pharmacy, Johannes Gutenberg-University Mainz, Staudinger Weg 5, D-55099 Mainz, Germany
The alkaloid pattern of cell and tissue cultures of the Indian medicinal plant Rauvolfia serpentina Benth. ex Kurz (Apocynaceae) has been extensively investigated in the past and more than 30 indole alkaloids were identified. It is noteworthy that 11 of these alkaloids were novel compounds found exclusively in R. serpentina cells cultured in vitro, including 6 alkaloids of the raumacline group [1]. All raumaclines were isolated after feeding of ajmaline to R. serpentina cell suspensions and therefore represent products of a biotransformation [2, 3].
Here we report the isolation and structural elucidation of 10-hydroxy-Na-demethyl-19,20-dehydro-raumacline (1), the first natural alkaloid of the rare raumacline type. The spectral data of the new compound (including results of 2D-NMR experiments) suggested a monoterpenoid indole alkaloid possessing unsubstituted a- and b-nitrogens and a structure similar to that of raumacline (2) [2] but bearing an additional hydroxyl group at C-10 and possessing a non reduced 19,20 bond (1). For studying the relative stereochemistry of 1, NOESY and ROESY measurements were performed. The analysis of these data allowed us to conclude that the substance (1) exists in solution as an equilibrium mixture of stereomeres in two stable interconvertible configurations with minimized energy e.g. E (major) and Z (minor) forms. Taking into account the structure of 1, we propose its biosynthetic formation from the Rauvolfia alkaloid vomilenine (3), which was detected in the extracts of R. serpentina hairy roots in fair yield. In contrast, each of the previously isolated raumaclines derives from ajmaline.
[1] J. Stöckigt, Biosynthesis in Rauwolfia serpentina Modern Aspects of an Old Medicinal Plant (1995) In: The Alkaloids (ed By G.A. Cordell) Vol. 47. Academic Press, New York,1995, pp. 115-172 (and literature cited therein).
[2] L. Polz, J. Stöckigt, H. Takayama, N. Ushida, N. Aimi, and S.-I. Sakai (1990) Tetrahedron Lett. 31:6693-6696.
[3] S. Endreß, H. Takayama, S. Suda, M. Kitajima, N. Aimi, S.-I. Sakai and J. Stöckigt (1993) Phytochemistry 32:725-730.
1 Department of Pharmacy, Division of Pharmacognosy, University of Athens, Panepistimiopolis Zografou, 15771 Athens, Greece
2 Institute of Botany, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11000 Belgrade, Yugoslavia
The genus Micromeria Bentham includes about 130 species wide spread in the Mediterranean region (1). In the Yugoslavian flora this genus is represented by 10 species seven of which are endemic (2).
In our framework on Micromeria we have studied the essential oils of four endemic species: M. thymifolia (Scop.) Fritsch, M. albanica (Griseb. ex K. Malý) Silic, M. dalmatica Bentham and M. pulegium (Rochel) Bentham.
The samples were gathered in the flowering period. The essential oils were obtained by hydrodistillation. The analyses of the oils were carried out using GC/MS. The identification of the compounds was based on comparison of their Kovats indices (KI), their retention times (RT) and mass spectra with those obtained from authentic samples and/or the NIST/NBS Wiley libraries (3).
The essential oils were found to be very similar in composition, oxygenated terpenes of the menthane type dominated in all the oils (> 50%).
The main components of the M. thymifolia essential oil were: isomethone (6.94%), pulegone (37.20%), piperitone + piperitone oxide (27.20%), piperitenone (6.27%) and piperitenone oxide (9.79%). Menthone was found in a small amount (0.46%). Isopulegone was detected in trace, while isomenthol was not present.
The oil of M. albanica contained menthone (4.49%), pulegone (7.77%), piperitone oxide 36.94%, piperitenone (9.97%) and piperitenone oxide (21.91%). Isomenthone was found in traces, while isopulegone, isomenthol and piperitone were not detected.
M. dalmatica oil was rich in pulegone (12.07%), piperitone (3.27%), and piperitenone (56.70%). Isomenthone (0.61%), isopulegone (0.52%) and piperitenone oxide (0.87%) were found in small amounts. Menthone was found in traces, while isomenthol and piperitone oxide were not detected.
The following compounds were present in the M. pulegium essential oil: menthone (4.26%), isomenthone (27.16%), isomenthol (2.57%), pulegone (4.02%), piperitone oxide (7.37%), piperitenone (1.20%) and piperitenone oxide (3.59%). Isopulegone and piperitone were not detected in the oil.
The similarity of the morphological characteristics of the investigated species as well as the quantitative and qualitative composition of their essential oils supports their classification in the section Pseudomelissa (4) of the genus Micromeria.
1. Diklic, N. (1974): Micromeria Bentham.. In: Josifovic, M. (ed.), Flore de la République Socialiste de Serbie, 6, 458-462. Académie Serbe des Sciences et des Arts, Belgrade.
2. Silic, C. (1979): Monographie der Gattungen Satureja L., Calamintha Miller, Micromeria Bentham, Acinos Miller und Clinopodium L. in der Flora Jugoslawiens, 172-262. Zemaljski Muzej BiH, Sarajevo.
3. Wiley, J. (1984): NBS Library.
4. Boissier, E. (1879): Flora Orientalis, 568-575 . Basileae, Genèvae & Lugduni
1Institute of Pharmaceutical Biology, University Freiburg, Stefan-Meier-Str. 19, 79104 Freiburg, Germany
2Escuela de Quimica and CIPRONA, Universidad de Costa Rica, San José, Costa Rica
3Faculdad de Farmacia and CIPRONA,Universidad de Costa Rica, San José, Costa Rica
Oyedaea verbesinoides DC (Asteraceae, tribe Heliantheae, subtribe Ecliptinae) (1) is a tree growing in Central America. Recently, diterpenes of the ent-kaurane type and sesquiterpene lactones have been found (1, 2). Reinvestigation of the aerial parts of the plant afforded nine diterpenes, four of which (1 - 3 and 8) were already known for this species. To the best of our knowledge, compound 7 has not been previously described. Structure elucidation was based on one and two dimensional NMR as well as ESI and CI-MS analysis. Interestingly, no sesquiterpene lactone could be detected in this plant material which was collected in Costa Rica as the previous one, but from a different area.
Ent-Kauranes are known for their inhibitory effects on rat uterus contraction (3). Therefore, studies are in progress to gain more information on which structural elements may influence this inhibitory activity.
1
R = CH2OH (ent-19-hydroxy-kaur-16-en)
2 R = CHO (ent-kaur-16-en-19-al)
3 R = COOH (ent-kaur-16-en-19-acid)
4 R1, R2 = H (ent-9 a-hydroxy-kaur-16-en-19-acid)
5 R1 = OAng, R2 = H (ent-15- b-angeloyloxy-9a-hydroxy-kaur-16-en-19-acid)
6 R1 = OTig, R2 = H (ent-15- b-tigloyloxy-9a-hydroxy-kaur-16-en-19-acid)
7 R1 = OAng, R2 = OH (ent-15- b-angeloyloxy-7a,9 a-dihydroxy-kaur-16-en-19-acid)
8 ent-9(11)-dehydro-kaur-16-en-19-acid 9 ent-kauran-16-b-ol
Stokes, S., Castro,V., Poveda, L., Papastergiou, F., Jakupovic, J. (1992) Phytochemistry 31, 2894-2894
Seaman, F., Bohlman, F., Zdero, C., Marby T.J., Diterpens of Flowering Plants, Springer Verlag (1990), 179,183,186,188
Campos-Bedolla, P., Campos, M.G., Valencia-Sánchez, A., Ponce-Monter, H., Uribe, C., Osuna, L., Calderón, J. (1997) Phytother. Res. 11, 11-16
a Institut für Pharmazie, Abt. Pharmakognosie, Universität Innsbruck, Innsbruck, Austria
b Crop & Food Research, Chemistry Dept., University of Otago, Dunedin, New Zealand
Polyacetylenes are characteristic compounds of the plant families Apiaceae, Araliaceae, Asteraceae, and some related families. Polyacetylenes in these higher plants usually possess a C17-chain, but some compounds with C18, C15, C14, C13 , C11, C10 are also known from taxa of the Apiaceae. As the biosynthesis of polyacetylenes in higher plants is based on oleic acid (C18H34O2), no polyacetylenes with a chain-length over C18 are currently known in trachaeophyta. Longer chain polyacetylenes, up to C45, have been reported from marine sponges (Faulkner, 2000). We now report the isolation of a C34-polyacetylene from the New Zealand plant Aciphylla scott-thomsonii Cockayne & Allan (Apiaceae, tribe Apieae). A. scott-thomsonii (giant speargrass) is a herb of up to 4 m height, guarded by fiercely sharp leaves and inflorescences, which is confined to subalpine regions in the southern part of the South Island of New Zealand. Methanolic extracts of A. scott-thomsonii yielded polyacetylenes 1-4. Known compounds (+)-falcarindiol 1, (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 2, and (+)-9(Z),17-octadecadiene-12,14-diyne-1,11,16-triol 1-acetate 3 were elucidated by 1H and 13 C NMR spectroscopy.
The structure of
compound 4 was established as
16-formyl-3,8,26,31-tetrahydroxy-tritriaconta-1,9(
Z),16(E),24(Z),32-pentaene-
4,6,27,29-tetrayne
by HR mass spectrometry and 1D- and 2D-NMR spectroscopy.
Compound 4 is of special interest since it represents the first polyacetylenic compound with a chain length of more than 18 carbon-atoms from a higher plant. Polyacetylenes in higher plants are synthesized from oleic acid (C18 H34O2). We therefore propose that the biosynthesis of 4 is achieved by an aldol condensation of two molecules of the aldehyde 5. The intermediate 5 is not known, but a related C 18-polyacetylene-aldehyde has been isolated from the Apiaceae Pastinaca sativa (Jones et al., 1966).
An aldol condensation as proposed for the biosynthesis of aciphyllal, proved to be involved in the biosynthesis of botryals, even C52-C 64 a -branched, a -unsaturated aldehydes isolated from the green alga Botryococcus braunii (Metzger & Casadevall, 1988, 1989).
Faulkner, DJ (2000): Nat. Prod. Rep. 17, 7-55.
Jones ERH, Safe S, Thaller V (1966): J. Chem. Soc. C, 1220-1221.
Metzger P, Casadevall E (1988): Tetrahedron Lett. 29, 2831-2834.
Metzger P, Casadevall E (1989): Phytochemistry 28, 2097-2104.
1Institut für Pharmazeutische Biologie, Heinrich-Heine-Universität Düsseldorf, Universitätsstrasse 1, D-40225 Düsseldorf, Germany
2Instituto de Quimica, Universidad Nacional Autonoma de Mexico, Circuito Exterior, Ciudad Universitaria, Apartado Postal 70213, Mexico, D. F., Mexico.
The genus Neurolaena (Asteraceae) is containing a total number of twelve Central American species (1). In contrast to the widely distributed N. lobata, which is known as remedy against several diseases (2), N. macrocephala is endemic to Veracruz, Mexico (1).
Recently, we reported the occurrence of ester derivatives of neurolenin-type sesquiterpene lactones in N. macrocephala (3). While N. lobata and N. cobanensis only contained isobutyric acid esters, N. macrocephala was the first species in which isobutyryl and 2-methylbutyryl ester side chains were also found (3). Later on such compounds were also found in N. oaxacana (4).
In continuation of our studies on sesquiterpene lactones from the genus Neurolaena, we now report the isolation and structure elucidation of new germacranolides of the neurolenin (1-2) and furanoheliangolide type (3-6). The furanoheliangolides 3 and 4 are of special interest, because of the a -oriented methyl group at C-4. Such compounds are unique in the genus, all other 4,5-dihydro-atripliciolides found so far contained a b-oriented methyl group at C-4. The structures of all compounds were elucidated using their GC/MS-1H- and 13C-NMR spectra including two dimensional experiments (COSY, HMQC, HMBC).
1. Turner, B.L. (1982) Plant Syst. Evol. 140, 119-139.
2. Giron, L.M., Freire, V., Alonzo, A. and Cáceres, A. (1991) J. Ethnopharmacol. 34, 173-187.
3. Passreiter, CM., Stoeber, S., Ortega, A., Maldonado, E., Toscano, R.A. (1999) Phytochemistry 50, 1153-1157.
4. Passreiter, CM., Sandoval-Ramirez, J., Wright, C.W. (1999) J. Nat. Prod.. 62, 1093-1095.
University of Münster, Institute of Pharmaceutical Biology and Phytochemistry, Hittorfstr. 56, D-48149 Münster, Germany
Extracts from the rhizomes of Cimicifuga racemosa are used in the therapy of menopausal complaints and premenstrual syndrome. Recently, a fraction enriched with cinnamic acid esters from a commercial available extract was shown to have an estrogenic effect in the MCF-7 test system. One particular compound of this fraction, fukinolic acid, was found to be one of the active constituents [1]. The aim of the present study was the isolation and characterization of hitherto unknown phenolic compounds from this active fraction.
The combination of different chromatographic steps (CC on SephadexÒ LH20 and MCIÒ-Gel, prep. HPLC on RP8 material) yielded cimicifugic acid D (1), fukiic acid (2), cimiciphenol II (3), petasiphenon (4) and cimiciphenon II (5), besides recently isolated cinnamic acid esters (fukinolic acid, cimicifugic acids A, B, E, F) [1]. The occurrence of compounds 1 and 3, previously detected by LC-MS analysis [2], have been unambiguously established here for the first time. Structure elucidation was performed using different spectroscopical methods such as ESI-MS, 1H-, 13C-NMR with HMBC, HMQC and NOE experiments. In analogy to the nomenclature of cimiciphenol II (3) and petasiphenon (4) (the latter was isolated from Petasites japonicus [3]), the new compound 5 is named cimiciphenon II.
1 Kruse S.O., Löhning A., Pauli G., Winterhoff H., Nahrstedt A. Planta Med. 1999; 65: 763 - 764
2 Hagels H., Baumert-Krauss J., Freudenstein J., Poster: 48th Annual Meeting of the Society for Medicinal Plant Research (GA), Zürich, 2000: P 1B/03
3 Yaoita Y., Kikuchi M., Phytochemistry 1994; 37: 1773 - 1774
Institute of Pharmacy, Department of Pharmacognosy, University Innsbruck, Austria
Uncaria tomentosa (Willd.) DC. (Rubiaceae), a woody vine which is better known as una de gato or cat´s claw , has been widely used by native Peruvian people to treat gastric ulcers, rheumatism, inflammations, tumors and also as a contraceptive [1]. Nowadays, this plant is not only of scientific but also of commercial interest. This fact becomes evident by more than 50 dietary supplement manufacturers in the United States offering cats claw products [2]. Up to now, a number of reports have been published on the chemical constituents of U. tomentosa, particularly on alkaloids [1-3]. Two chemotypes of the plant are known to exist, one producing pentacyclic, and the other one tetracyclic oxindole alkaloids. In many cats claw products, however, a mixture of both alkaloid groups can be found [2].
The aim of this study was to develop a rapid and efficient method for the isolation of penta- and tetracyclic oxindole alkaloids from U. tomentosa and an improved HPLC-MS method for the quantification of these compounds.
The separation of all eight isomers was successfully achieved by high-speed counter-current chromatography (HSCCC) using a two-phase solvent system composed of n-hexane : ethylacetate:MeOH:H 2O (60:40:25:100). The lower, aqueous phase was used as mobile phase for a pH-modulated stepwise elution, starting with a pH-value of 5.2 and ending with pH 3.9. The rotation speed was kept constant at 950 rpm, detection of compounds was performed by TLC. Seven of the eight alkaloids could be isolated in >97 % purity. The optimised HSCCC conditions allowed reproducible separation of oxindole alkaloids from U. tomentosa (n=3) and were also applicable in scale-up experiments for preparative isolation of the oxindole alkaloids using a coil with a capacity of 325 mL.
Analysis of alkaloids from U. tomentosa was performed by HPLC-MS using an electro spray ionization (ESI) interface. Baseline separation of all alkaloids was achieved using a Merck, LichroSpher 60® RP Select B column (250 x 4 mm) and a solvent gradient composed of ammonium acetate (10 mM, pH 6.7) and acetonitrile at a flow rate of 1 mL/min. The system was operated at room temperature, the detection wavelength was 245 nm, and the injection volume 10 µL. MS detection was performed in the positive ionization mode at a spray voltage of 4.5 kV and a capillary temperature of 150 °C. The sheath gas flow was 45 psi and the LC flow was split 1:5. When 20 V were applied as in-source collision induced dissociation (CID) voltage, the signals of the quasi molecule ions [M+H]+, m/z 369 for the pentacyclic and m/z 385 for the tetracyclic alkaloids, dominated the mass spectra.
Applying a sufficient high in-source CID (50 V), however, reproducible fragmentation of the [M+H] + ions was observed which allowed to distinguish between the two isomers of the normal series and isomers of the allo- or epiallo series.
[1] Kenneth Jones (1995) In: Cats Claw, Healing vine of Peru. Sylvan Press, Seattle
[2] K. Keplinger et al. (1999) J Ethnopharmacol 64, 23-34
[3] K.H. Reinhard (1999) J Altern Complement Med 5, 143-151
1 Institute of Pharmacy, Department of Pharmacognosy, University Innsbruck, Austria
2 Department of Pharmaceutical Sciences, University of Florence, Italy
The roots of Cephaelis ipecacuanha (Brot.) A. Richard (Rubiaceae) have been used in therapy since the beginning of the 17th century as an emetic and expectorant, and were recommended as early as in the 18th century for the treatment of dysentery. The medicinal value of this drug rests mainly in its content of emetine and cephaeline, which are in particular responsible for its pharmacological effects [1]. Nowadays several ipecacuanha formulations are listed in US and European pharmacopoeias. They are particularly used for their emetic properties in hospitals as a first-aid medicine after intoxications and are considered more effective than gastric lavage, especially for children [2].
In the Pharmacopoeias quantitative determination of the alkaloids is performed by titration [3] or by a more complex spectrophotometric method, which involves a liquid-liquid chromatographic fractionation [4]. In the literature a number of TLC and normal-phase or ion-pair HPLC methods are described [5-7].
In this study capillary zone electrophoresis (CZE) was applied for sensitive qualitative analysis as well as accurate and reproducible quantitative determination of cephaeline and emetine in extracts and several pharmaceutical formulations.
Baseline separation within 10 min was achieved by CZE using a fused silica capillary (67.5 cm x 75 µm ID) and a citric acid (50 mM) / phosphate (100 mM) buffer solution (pH 4.4) containing 2.5 % of methanol. The voltage was kept constant at 25 kV, the temperature at 20 °C. Injection was performed in the pressure mode (50 mbar x 3 sec) and for UV detection a wavelength of 205 nm was applied. The impact of the electrolyte composition (concentration, pH, additives), the applied voltage and the thermostating temperature on the resolutions of adjacent peaks will be discussed.
Quantitative determination of cephaeline and emetine in a tincture, fluid extract and in Ipecac Syrup [4,8] was performed using papaverine as internal standard. Most samples could be injected without any special pretreatment. Before quantitative analysis of Ipecac Syrup, however, alkaloids had to be separated from other components by a simple procedure using ion exchange solid phase extraction. The standard deviations for all samples studied were < 5%. Recovery rates for emetine were between 99,79 and 100,25 %.
[1] Shamma , M. (1972) in The Isoquinoline Alkaloids, p.426. Academic Press, New York.
[2] A.H. Abdallah, A. Tye (1961), Am. J. Dis. Child. 113, 571
[3] European Pharmacopoeia, 3rd Ed., Supplement 2000, 851-52
[4] USP 24, 2000, 913-915
[5] R. Verpoorte et al. (1974), J. Chromat. 100, 227-230
[6] Lj. Kraus et al. (1985), Dtsch. Apoth. Ztg. 125, 863-864
[7] S.J. Bannister et al.(1979), J. Chromat. 176, 381-390
[8] Farmacopea Ufficiale della Repubblica Italiana, 1991, 187
1Institute of Pharmacy, Department of Pharmacognosy, University of Innsbruck, Austria
2Institute of Organic Chemistry, University of Innsbruck, Austria
Previous phytochemical investigations of the alpine plant Leontopodium alpinum Cass. (Asteraceae) resulted in the isolation of flavonoids [1], phenolic acids [2], terpenes [3] and one chromane derivative [4] as well as isocromene, modhephene, caryophyllene [5] and bisabolane [6] type sesquiterpenes. We here report the isolation and structure elucidation of a number of new compounds: 1-(2-[1-(hydroxymethyl)vinyl]-3-[ b-D-glucosyloxy]-2,3-dihydrobenzo[b]furan-5-yl)-1-ethanone (1), the lignane derivative [(2R*,3S,4S)-4-(3,4-dimethoxybenzyl)-2-(3,4-dimethoxyphenyl)tetrahydro-3-furanyl]methyl (E)-2-methyl-2-butenoate (2), the silphiperfolen-type sesquiterpene [(1R*,3aS,6R)-1,3a,6-trimethyl-1,3a,4,5,5a,6,7,8-octahydrocyclopenta[ c]pentalen-2-yl]methyl acetate (3) and the obliquine-type coumarin 8-hydroxy-2-isopropenyl-2,3-dihydro-7 H-[1,4]dioxino[2,3-g]chromen-7-one (4). Additionally, the coumarin obliquin (5), already known as a constituent of Ptaeroxylon obliquum [7], was isolated. Isolation of compounds 1-5 was performed by consecutive silica gel and Sephadex LH-20 column chromatography, structural elucidation by HR-MS and 1- and 2-D NMR spectroscopy.
[1] Tira, S., Galeffi, C., DiModica, G. (1970): Experentia 26, 1192.
[2] Hennessy, D., Hook, I., Sheridan, H., McGee, A. (1989): Phytochemistry 28, 489-490.
[3] Bicchi, C., Frattini, C., Nano, G.M., Tira, S. (1979): Relata Tech. 11, 64.
[4] Comey, N., Hook, I., Sheridan, H., Walsh, J. (1997): J. Nat. Prod. 60, 148-149.
[5] Comey, N., Grey, A.I., Hook, I.L., James, P., Sheridan, H. (1999): Phytochemistry 50, 1057-1060.
[6] Stuppner, H., Ellmerer-Müller, E.P., Ongania, K-H., Mayer B. (1998): 46th Annual Congress of the Society for Medicinal Plant Research, Vienna 1998.
[7] Dean, F.M., Taylor (1966): J. Chem. Soc. 114.
1Pharmacognosy Department, Faculty of Pharmacy, Helwan University, Cairo, Egypt
2School of Pharmaceutical Sciences, University of Shizuoka-Shizuoka 422-852, Japan
3Chemistry of Natural Products Department, National Research Centre, Dokki,Cairo, Egypt
The methanolic extract of the leaves of Agave lophantha was concentrated and diluted with acetone to afford a crude saponin mixture. Our pharmacological evaluation of the mixture indicated that it exhibited significant anti-inflammatory and anti-ulcerative activities. Phytochemical study carried out by us has resulted in the isolation of two new saponins 1 and 2, after repeated CC and HPLC.
Saponin 1 (C50 H82 022) was identified as spirostanol saponin from its spectral (IR, NMR) features. The 1 H and 13C NMR resonances, due to the aglycone, were assigned by the combined use of 1D and 2D (1 H-1H COSY, HMQC, HMBC) NMR techniques. Comparison of the assigned resonances with the reported data of related compounds, indicated that 1 was a glycoside of smilagenin [(25R)- 3 b-hydroxy-5b-spirostan]. Acid hydrolysis afforded the aglycone and the sugar components D-apiose, D-glucose and D-galactose identified by TLC and GC. The spin system of each monosaccharide unit was analysed from the combined use of HOHAHA difference, NOE difference and 1H-1H COSY NMR spectra. The 13C NMR resonances of each sugar unit were identified by HMQC and confirmed by HMBC experiments. The site of attachment of the saccharide part and the interglycosidic linkages were established from the interresidue NOEs and long-range HMBC correlations. Based on the above spectral analyses, the structure of 1 was elucidated as (25R)-5 b-spirostan-3b-0l-3-0-{ b-D-apiofuranosyl (1®4) b-D-glucopyranosyl(1®3)[ b-D-glucopyranosyl(1®2)] b-D-galactopyranoside}.
Saponin 2 (C 50H 82 0 22) exhibited the spectral features of furanostanol glycoside. On the basis of its assigned 1 H and 13C NMR resonances and by comparison with the reported data of related compounds, the aglycone of 2 was identified as (25S)- 5b- furost-20 (22)-ene-3 b-26 diol substituted at C-3 and C-26 positions. Acid hydrolysis afforded the aglycone smilagenin and the sugars D-xylose, D-glucose and D-galactose. The identity of the monosaccharide units, their sequencing and linkages were determined following the same protocol used in 1. Based on that, 2 was assigned the structure of 26-O-b -D-glucopyranosyl-(25R)-5b-furost 20(22) ene - 3b, 26-diol-3-O-{ b-D-xylopyranosyl(1®3)-[ b-D-glucopyranosyl(1®2)] b-D-galactopyranoside}.
1 2
Api= b-Dapiofuranosyl;Glc= b-Dglucopyranosyl; Xyl=b-D xylopyranosyl; Gal= b - D - galactopyranosyl
1Institute of Pharmacy, Pharmaceutical Biology, Freie Universität Berlin, Königin-Luise-Str. 2+4, D-14195 Berlin, Germany
2Department of Chemistry, Faculty of Pharmacy, University of Istanbul, 34452 Istanbul, Turkey
The genus Salvia (Lamiaceae) is represented by 90 species in the flora of Turkey, including 44 species that are endemic to this area (1). Salvia species are used in folk medicine for their antibacterial, antioxidant, antidiabetic, antitumour and antituberculotic activities. Chemically, a large variety of diterpene structures have been found and their frequent occurrence in the lamiaceae has been used in chemotaxonomic studies. Continuing our investigation on the diterpene patterns of Salvia species of Turkey, we present here the isolation and structural charaterization of 5 diterpenoid metabolites from S. cilicia, including the two new abietane derivatives 1 and 2.
A combination of column and preparative thin layer chromatography led to the isolation of 7-hydroxy-12-methoxy-20-nor-abieta-1,5(10),7,9,12-pentaen-6,14-dione (1) and abieta-8,12-dien-11,14-dione (12-deoxyroyleanone) (2 ) from the acetone extract of the roots of the titled plant. Their structures have been established on the basis of spectroscopic evidence.
These metabolites were accompanied by the known diterpenoids, ferruginol (2), the rarely found inuroyleanol (11,14-dihydroxy-12-methoxy-abieta-8,11,13-trien-7-one (3), and the tentatively identified Salvia-specific cryptanol (4).
1. Davis, P. H. (1982). Flora of Turkey and East Aegean Islands, Vol 7, Edinburgh University Press, p. 400
2. Cambie, R. C., Madden, R. J. & Parnell, J. C. (1975). Helv. Chim. Acta, 58, 1921
3. Bhat, S. V., Kalyanaraman, P. S., Kohl, H., De Souza, N. J. & Fehlhaber, H. W. (1975) Tetrahedron, 31, 1001
4. Ulubelen, A., Topcu, G. & Terem, B. (1987). Phytochemistry, 26, 1534.
1Department of Cell Biology & Genetics University of Lagos, Akoka, Lagos, Nigeria
2Department of Biochemistry Nigeria Institute of Medical Research Yaba, Lagos, Nigeria
3Department of Chemistry, University of Lagos, Akoka Yaba, Lagos, Nigeria
A medico-ethnobiology study of the Igbos was carried out. Field survey, data collection, phytochemical and pharmacological tests were used . Dioscorea alata, Xapthosoma sagittifolia, Musa sapientum, Garcinia kola and Carica papaya are used for diabetes (2,3,4). Vernonia amygdalina, Azadirachta indica and Morinda lucida (4) are used for malaria. Vernonia amygdalina is also used for diabetes while A. indica oil is applied as a remedy for wounds, ulcers, skin diseases and jaundice. These plants have not shown acute toxicity, but toxicity test on pregnant rats show that 300 mg/kg of A. indica extract prevented pregnancy and was lethal to the litters (1). The appearance of an IR band at 1625 cm-1 indicates a C=C stretch of an unsaturated bond which is very reactive. This may explain toxicity at high dosage. In addition, tirucallol synthesized from A. indica resembles estradiol. Estradiol (1.0 mg/rat) and butin 20mg/rat were reported to have antiestrogenic activities in female rats when treated for 5 days (5). The mode of application, dosage, pictures, IR spectrum and structure will be presented.
R.K. Tita and P.G.C. Odeigah (1999). The Book of Abstracts 2000 years of Natural products Research. Amsterdam - The Netherlands, p. 781
P.G.C. Odeigah et. al. (1999) Diabetes International vol. 9: 71-73
Iwu et. al. (1990) J. Pharm Pharmacol 42: 290-2
F. Ilonzo (1995) You and Your health with Phytomedicines, p.26
Bhargava, S.K. (1986) Journal of Ethnopharmacology 18 95-101.
Department of Pharmacognosy, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran
Anthraquinone metabolites occur in various plant families. Frangula is one of the plant genera belonging to the family of Rhamnaceae, which because of the better taste and also the less stimulant effects on the gastrointestinal tracts within oral administration, have extensively uses in treatment of constipation as different dosage forms. Frangula grandifolia Fish. has previously been reported to grow in the northern region of Iran (1). The aerial parts of the plant such as stem bark, leaves and fruits contain 1,8-dihydroxyanthraquinones like emodin, chrysophanol, physcion and frangulin. In addition, commonly phytochemical tests indicated the presence of secondary metabolites including flavonoids, tannins and on the other hand the absence of alkaloids, saponins, cardiac glycosides and sennosides. The anthraquinones mentioned were identified by UV, FT-IR, H-NMR, C-NMR and MS, the total amount of 1,8-dihydroxyanthraquinones were determined according to the DAB 9 procedure (2) and estimated to be 4.50%, 2.00% and 0.65% in dried stem bark, fruits and leaves, respectively.
1. Rechinger, K.H.: Flora Iranica, Rhamnaceae, No.125, Akademische Druck-u. Verlagsanstalt, Graz (1977).
2. DAB 9 Kommentar, Wissenschaftliche Verlagsgesellschaft mbH, Stuttgart (1988).
1Department of Chemistry, Faculty of Science, El-Minia University, El- Minia 61519, Egypt
2Department of Pharmacy, Division of Pharmacognosy, University of Athens, Panepistimiopolis Zografou, 157 71 Athens, Greece
3Technical University Budapest, H-111 Budapest, Hungary
The genus Achillea (Compositae) comprises 100 species distributed worldwide [1]. Aerial parts of various species of this genus, especially A. millefolium, are widely used in folk medicine for fevers, common cold, essential hypertension, amenorrhoea and especially for thrombotic conditions with hypertension, including cerebral and coronary thromboses [2,3].
Several phytochemical studies exhibited that the main constituents of the genus are sesquiterpenes lactones and flavonoids. Achillea ligustica All., a member of the A. nobilis group, is found in the Mediterranean region and belongs to the section Millefolium.
As part of our work on isolation and identification of constituents from the genus Achillea [4-5], we report here the isolation of two new sesquiterpene lactones 4,10-a-dihydroxy-3- b-chloro-1,2-b-epoxy-guaia-11(13)ene-6 a,12-olide (1), achinone (2) and three known compounds, matricarin, 2,6-dimethyl-octa-1,7-diene-3,6-diol and 5-hydroxy-5,6-secocaryophyllen-6-one from the aerial parts of A. ligustica. The structures were established by means of 1D and 2D NMR experiments (1H, 13C, 1H-1H COSY, HMQC, and HMBC)
|
|
|
|
1 |
2 |
D DJ Mabberley, The Plant-Book, 2nd edition, Cambridge University Press (1997).
CA Newall et al., Herbal Medicines A Guide for Health-care Professionals, The Pharmaceutical Press (1996).
RF Chandler et al., J. Pharm. Sci., 58 (1969) 938.
AA Ahmed et al., Phytochemistry, 29 (1990) 1322.
O Tzakou et al., Biochem. Syst. Ecol., 23 (1995) 569.
1 Institute of Pharmacognosy, University of Vienna, Centre of Pharmacy, Althanstr. 14, A-1090 Vienna, Austria
2 Institute of Pharmaceutical Chemistry, University of Graz, Schubertstr. 1, A-8010 Graz, Austria
Herbal remedies from species belonging to the Achillea millefolium group are widely used in folkmedicine. The most important indications are gastrointestinal disorders and different women`s diseases, based on the antiphlogistic, spasmolytic, choleretic, hemostyptic and antimicrobial properties of the plant [1]. These effects are due to the content of essential oil, sesquiterpenes, flavonoids, caffeoylacid derivatives, coumarins etc. All taxa are collected for traditional use, although they show differences not only in morphology but also in chemistry. The sesquiterpenes, which are subject of the presented work, are not only important for the antiphlogistic effect, but show also chemotaxonomic relevance [2].
In continuation of our investigation of the sesquiterpenes from Achillea pannonica [3] we now report on the isolation and structure elucidation of two further sesquiterpenes, 11,13-dehydro-desacetylmatricarin (1) and a-longipin-1-en-2-on (2).
The plant material was collected in Falkenstein and Buschberg, Lower Austria and the ploidy level was determinated as octoploid by flowcytometry. The dichloromethane extract of the air dried flowerheads was separated by column chromatography (silica gel, dichloromethane-acetone), further separation and purification was performed by HPLC (RP-8, methanol-water) and small sized column chromatography (silica gel, dichloromethane-acetone). The structure of both sesquiterpenes was elucidated by MS (EI and CI), IR and NMR (1H, H,H-COSY, HSQC, HMBC, NOE-experiments).
Together with the structures of two sesquiterpenes previously reported [3], there are now four different types of sesquiterpenes described for Achillea pannonica: guaianolide (1), longipinane (2), germacrane [4] and farnesane derivatives [3], representing a great variety compared with the other taxa of the Achillea millefolium group and confirming the already observed trend, that Achillea taxa of higher ploidy seem to be characterized by a greater number of different sesquiterpene skeletons than taxa on a lower ploidy level.
|
|
|
Willuhn G. in Wichtl M. (1997) Teedrogen und Phytopharmaka, Wissenschaftliche Verlagsges. mbH Stuttgart, 3. Auflage: 395-399
Kubelka W., Kastner U., Glasl S., Saukel J., Jurenitsch J. (1999) Biochem. Sys. Ecol. 27, 437-444
Werner I., Glasl S., Presser A., Haslinger E., Jurenitsch J., Kubelka W. (2000) Poster Nr. P1A/49 at the 48th GA Congress 2000
Sosa S., Tubaro A., Kastner U., Glasl S., Jurenitsch J., Della Loggia R., Planta medica, in press
1 Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, 4099-003 Porto, Portugal
2 Centro de Estudos de Química Orgânica, Fitoquímica e Farmacologia da Universidade do Porto, Faculdade de Farmácia, 4050-047 Porto, Portugal
3 Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand
The plants from the large tropical genus Garcinia (Guttiferae) are widely used in traditional medicine and are known to contain a great variety of phenolic compounds including xanthones, biflavonoids and benzophenones. Only few reports on their triterpene content are available indicating that they consist mainly of the friedelane and lanostane (cicloartane derivatives) groups.
To the best of our knowledge only three prenylated xanthones have been isolated from Garcinia speciosa and the inhibitory properties of these xanthones on acidic sphingomyelinase has been also reported [1].
We here report the phytochemical investigation of G. speciosa Wall. from Thailand. The chloroformic extract of its bark afforded, besides six known steroids ( b-sitosterol, 3-O-acetyl-b-sitosterol, stigmasterol, 3-O-acetylstigmasterol, 24-ethyl-3-oxo-colest-4-en and 24-ethyl-3-oxo-colest-4,22-dien) three triterpenes, friedelin, 3 b-hydroxy-23-oxo-9,16-lanostadien-26-oic acid (1)[2 ] and its acetate derivative (2) which is a new natural compound. The structure of 1 was also confirmed by X-ray crystallography.
1. C. Okudaira, Y. Ikeda, S. Kondo, S. Furuya, Y. Hirabayashi, T. Koyano, Y. Saito, K. Umezawa, J. Enzyme Inhibition (2000), 15, 129-138.
2. V. Rukachaisirkul, A. Adair, P. Dampawan, W.C. Taylor, P.C Turner, Phytochemistry (2000) 55, 183-188.
Acknowledgements: We thank FCT (ID nº 226/94), POCTI (QCA III), FEDER and PRAXIS XXI for support.
Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria
! Hacettepe University, Faculty of Pharmacy, Department of Pharmacognosy, 06100 Ankara, Turkey
Tunicates can be considered as a source of numerous natural products, many with significant human health benefits [1,2]. However, there are only limited data on Mediterranean tunicates. The aim of the present work was to characterize the chemical composition of Styela sp. and Phallusia sp. from the Eastern Mediterranean. Sterols, volatiles and lipids were isolated from the lipophylic extracts from both tunicates and identified by means of GC, GC/MS and 1 H-NMR. The sterol composition of the both organisms appeared to be similar with the exception of the concentrations of 5a -stanols. The main peaks of the GC curves of the sterol mixtures from the samples corresponded to cholesterol and its saturated analogue, followed by 24-methyl-cholesta-5,22-dien-3b-ol and 24-ethyl-cholest-5-en-3b-ol together with their saturated analogues. Both tunicates were characterized with the presence of sterols with 22Z-double bond. In the volatiles were found significant amounts of chlorinated compounds (phenols in Styela sp. and hydrocarbons in Phallusia sp.). The fatty acid composition of triacylglycerols and phospholipids of two tunicates showed significant differences, which may have chemotaxonomical importance for future studies.
Davidson, S. B. Chem. Rev. 1993, 93, pp. 1771-1791.
Faulkner, D. JNP Reports 1995, 12, pp. 256-258.
a Institut für Pharmazie, Abt. Pharmakognosie, Universität Innsbruck, Innsbruck, Austria
b Crop & Food Research, Chemistry Dept., University of Otago, Dunedin, New Zealand
Recently the new irregular diterpenes anisotomenoic acid 1 and anisotomene-1-ol 2 were discovered in the New Zealand Apiaceae Anisotome flexuosa (van Klink et al., 1999). A detailed study of this species and A. haastii yielded two more compounds 4-5 with the same basic carbon skeleton (van Klink et al., 2001). We now report the isolation and structure elucidation of 16-hydroxyanisotomenoic acid 3 from A. flexuosa and of 13,14-dihydroanisotom-12E-ene-1,14-diol 6 and 14-methoxy-13,14-dihydroanisotom-12 E-ene-1-ol 7 from A. haastii. Compounds 6-7 differ from all known anisotomene derivatives in the position of the side-chain double bond. Structure elucidations were performed by HR mass spectrometry and 1D- and 2D-NMR spectroscopy.
Extracts of A. pilifera yielded known compounds 1-2, 6,7-dimethoxycoumarin, falcarindiol, chlorogenic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and luteolin 7-O- a-L-rhamnosyl(1®6)- b-D-glucoside, and the new sesquiterpenoid 8-O-senecioyl-6 b,8a,11-trihydroxygermacra-1(10)E,4E-diene 8.
Conformational searching and molecular modeling was carried out on the four possible C-6/C-8 configurations, assuming the normal 7 b-isopropyl stereochemistry. Only the 6 b,8a-stereoisomer 5 gave a good match of predicted coupling constants (agreement £ 2 Hz) and NOE interactions (strong for distances < 3 Å) with the experimental data. The predicted predominant ring conformation of 5 was the crossed/boat conformation sensu Appendino et al. (1986).
Appendino G, Valle MG, Gariboldi P (1986): J. Chem. Soc. Perkin Trans I, 1363-1372.
van Klink JW, Barlow AJ Perry NB, Weavers RT (1999): Tetrahedron Lett. 40, 1409-1412.
van Klink JW, Barlow AJ, Benn MH, Perry NB, Weavers RT (2001): Aust. J. Chem. (in press).
Department of Pharmaceutical Sciences and Chemistry, University of Antwerp, 2610 Antwerp, Belgium
Chinese medicinal herbs are increasingly used in the Western world. However, information on their phytochemical constituents, and certainly which of these contribute to the biological activity, is often lacking. Since in herbal medicine, and also in the case of the two plants investigated here, Wedelia chinensis (Osb.) Merr. (Compositae) and Kyllinga brevifolia Rotb. (Cyperaceae), mainly aqueous preparations are used, the present communication focuses on the characterisation of known and new hydrophilic compounds from these plants, the structure of which was elucidated by 1D- and 2D-NMR methods and mass spectrometry.
A series of caffeic acid derivatives (3,5-, 3,4-, and 4,5-dicaffeoyl-quinic acid), the new compound b,3,4-trihydroxyphenethyl-O-[ b-apiofuranosyl-(1®4)- a-rhamnopyranosyl-(1®3)] -(4-O-caffeoyl)- b-glucopyranoside (wedelosin), as well as quercetin 3-O- b-glucoside, kaempferol 3-O-b-apiosyl-(1-2)- b-glucoside, and astragalin (or kaempferol 3-O- b-glucoside) were isolated from Wedelia chinensis. Wedelosin exhibited an inhibitory activity on both the classical and the alternative activation pathway of the complement system (IC50 = 18.9 and 50.3 mg/ml, respectively). Also quinic acid derivatives and certain flavonoids are known for their complement-inhibiting properties [1]. The complement system plays a role in a variety of inflammatory and degenerative diseases, and the presence of complement-inhibiting compounds may at least in part explain the traditional use of Wedelia chinensis against e.g. rheumatoid arthritis [2].
Kyllinga brevifolia yielded two known flavonoid glycosides (kaempferol 3-O- b-apiosyl-(1-2)-b-glucoside and isorhamnetin 3-O-b-apiosyl-(1-2)- b-glucoside), and a new quercetin triglycoside (quercetin 3-O -b-apiofuranosyl-(1 ®2)-b-glucopyranoside 7- O -a-rhamnopyranoside). The latter compound showed a moderate antiviral activity (102 reduction factor of the viral titer of Herpes simplex virus type 1 at 100 mg/ml). In traditional medicine Kyllinga brevifolia is used for various indications, including treatment of viral diseases such as common cold and influenza [2]. Nevertheless, compounds isolated up to now only showed a moderate antiviral activity. The antiviral activity observed for the crude hydrophilic extract of Kyllinga brevifolia may be related to the presence of still unidentified proanthocyanidins and polysaccharides.
|
|
Wedelosin
1. Huang Y. Ph. D. Thesis, University of Antwerp, Belgium (2000). 2. Pieters, L., De Bruyne, T. and Vlietinck, A. J. In Immunomodulatory Agents from Plants, Wagner, H., Ed., Birkhäuser Verlag, Basel, pp. 137 160, 1999. |
1 Dipartimento di Scienze Farmaceutiche, Università degli Studi di Padova, via Marzolo 5, Italy
2 Dipartimento di Scienze Farmaceutiche, Università degli Studi di Salerno, Penta di Fisciano (SA), Italy
3 Dipartimento di Biologia, Università degli Studi di Padova, via Bassi 58/B, Italy
In the course of the establishment of a protocol for the regeneration of Haplophyllum patavinum, an endangered plant of the Rutaceae family, the biosynthetic capabilities of in vitro cultures were evaluated. We previously reported on the identification of various coumarins [1], lignans [2] and furoquinoline and quinolone alkaloids [3].
In continuation of our phytochemical investigations of H. patavinum we now report the isolation of three arylnaphthalene lignan glycosides biosynthesized by shoot cultures.
A new arylnaphthalene lignan glycoside (1) was characterized. The structure of 1 was established as ( -)-diphyllin 4-O-[ b-D-apiofuranosyl(1 ®4) b-D-xylopyranosyl(1 ®2) b-D-apiofuranoside], patavine, on the basis of detailed spectral analysis by UV, 1D- and 2D-NMR (1H ,13C, HMBC, HSQC) and ESI-MS. Majidine and arabelline, two diphyllin diglycosides, originally reported from H. buxbaumii [4] were also isolated and identified.
1
The three lignan glycosides were also present in the medium. This is an encouraging result, since excretion from the cells and accumulation in the spent medium is regarded to be a prerequisite for the large-scale production.
Research on biological activities of the isolated compounds are in progress.
Filippini R., Piovan A., Innocenti G., Caniato R., and Cappelletti E.M. (1998) Production of coumarin compounds by Haplophyllum patavinum in vivo and in vitro . Phytochemistry 49 (8), 2337-2340.
Innocenti G., Piovan A., Filippini R., Caniato R., Piacente S. Cappelletti E.M. (1998) International Symposium Antitumour Products from Higher Plants. Paris 8-10 January.
Puricelli L., Innocenti G., Delle Monache G., Caniato R., Filippini R., Cappelletti E.M. In vivo and in vitro production of alkaloids by Haplophyllum patavinum (2001) Nat. Prod. Letters Submitted.
Al-Abed Y., Sabri S., Zarga M.A., Shah Z., and Rahman A. (1990) Chemical constituents of the flora of Jordan, part V-B. Three new arylnaphthalene lignan glucosides from Haplophyllum buxbaumii. J. Nat. Prod., 53, 1152-1161.
1
2