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Iridoids are a class of secondary metabolites found in a wide variety of plants. Iridoids are typically found in plants as glycosides, most often found to glucose. The genus Stachys L. is one of the largest genera of the Lamiaceae family, containing iridoids. Aim The aim of this study was the isolation and identification of iridoids from the aerial parts of Stachys byzantina K. Koch and Stachys germanica L. Methods For the isolation and identification of the iridoids, different chromatographic methods (NP-TLC, CPC and RP-HPLC) were used. The structures were established by one- and two-dimensional NMR and mass spectrometry, also. Results Iridoids (aucubin, harpagide, ajugoside and harpagoside) were isolated and identified by combination of different chromatographic methods from S. byzantina and S. germanica. Conclusion Stachys species may also be used as a potential source of iridoids.

Grapefruit (Citrus paradisi) seeds are increasingly applied for medicinal purposes, especially due to their antimicrobial and immunostimulant features, to treat or prevent common cold. However, the scientific background of these uses has not been established. To date, only in vitro studies support the antimicrobial effect of the seeds, and there are no data on the systemic antibiotic or other clinical effects. Moreover, recent data refer to the adulteration of grapefruit seed products with synthetic antiseptics which may be an explanation for the antimicrobial efficacy of some formulations. The aim of our study was to develop a simple thin-layer chromatography- based method for the quality control of grapefruit seed extracts. Besides the analytical procedure, we report a quick and simple method for the isolation of limonoids from grapefruit seeds. Limonoids are characteristic compounds of the seeds; hence, the detection of these marker compounds may be the basis of grapefruit identification. Further, a th...

Two new 2,2-dimethyl-chromene derivatives and the new pentaisovaleryl sucrose were isolated from the roots of Artemisia asiatica together with the known 1-(5-acetyl-2,4-dihydroxy-phenyl)-3-methyl-butan-1-one, 6-hydroxytremetone and ( E )-coniferyl aldehyde. The structures of the compounds were elucidated by extensive 1D ( 1 H, 13 C and JMOD) and 2D ( 1 H– 1 H COSY, HSQC, HMBC and NOESY) NMR studies and APCIMS and HRESIMS investigations. 6-Acetyl-3,7-dihydroxy-2,2-dimethyl-chromene and 6-acetyl-3-hydroxy-2,2-dimethyl-chromene are biogenetically related to 1-(5-acetyl-2,4-dihydroxy-phenyl)-3-methyl-butan-1-one and 6-hydroxytremetone as all having isoprenoid acetophenone origin. All the isolated compounds were described for the first time from this species; interestingly none of them are related to the metabolites identified from the extracts of the aerial part of the plant.

Ethnopharmacological relevance Oxybaphus nyctagineus (Michx.) Sweet has traditionally been used by several Native American tribes predominantly as a topical anti-inflammatory and analgesic agent. Aim of the study To evaluate the antioxidant, analgesic and anti-inflammatory activity of the extracts prepared from the aerial parts of Oxybaphus nyctagineus and to characterize the major chemical constituents of the bioactive extracts. Materials and methods Crude polar and apolar extracts (PCE and ACE) of the herb of Oxybaphus nyctagineus were prepared and tested in the models of the CFA-induced hyperalgesia in rat knee and carrageenan-induced paw edema in rat. To identify the active compounds, subfractions were prepared by column chromatography and subjected in vitro assays, such as antioxidant assays (DPPH, peroxynitrite (ONOO−) scavenging), and the LPS-induced IL-1β release test in human monocytes. Preparative HPLC was employed for the isolation of active substances, while phytochemical analysis was performed by mean of LC–MS/MS and NMR. Results The topically administered PCE and ACE of Oxybaphus nyctagineus demonstrated a significant analgesic and anti-inflammatory effect in the inflammation animal models. The subfraction A4 of ACE and the subfraction P5 of PCE considerably inhibited the LPS-induced IL-1β release in human monocytes, while the strongest activity was localized in the subfraction P5 in the antioxidant assays. The HPLC–MS/MS and NMR analysis revealed that 6-methoxyflavonol diglycosides, namely patuletin-3-O-robinobioside (1), 6-methoxykaempferol-3-O-robinobioside (2), spinacetin-3-O-robinobioside (3), and hydroxy-polyenoic fatty acids, namely corchorifatty acid B (4), 9-hydroxy-10E,12Z,15Z-octadecatrienoic acid (9-HOT acid) (5), and 9-hydroxy-10E,12Z-octadecadienoic acid (9-HOD acid) (6) were present in PCE, and in ACE as major compounds. Conclusion The results of this study established a pharmacological evidence for the traditional use of Oxybaphus nyctagineus as an anti-inflammatory agent used topically, and provided data on its phytochemical composition for the first time.

Ginkgo biloba L. (Ginkgoaceae) is one of the oldest trees on earth. The medicinal use of its seeds and leaves has been a tradition for thousands of years. The standardized extract (known as EGb 761) contains several biologically active components, among them are terpenes and flavonoid glycosides that are responsible for the pharmacological activities of Ginkgonis folium. According to European Union herbal monographs (EUHM), the leaves of Ginkgo are recommended for the treatment of dementia, cerebral vascular insufficiency, and disorders of the peripheral circulation. The aim of our work was to analyze volatile constituents of Ginkgo leaf. Leaves of 3 Ginkgo trees were analyzed; 2 of which grow in the Medicinal Plants Garden (young trees A and B) and 1 at the Botanical Garden (old tree C) in Bratislava. The leaves were collected in 2014. The essential oil was isolated and quantified using hydrodistillation according to European Pharmacopoeia (Ph. Eur.) The volatile constituents of Ginkgonis folium were evaluated qualitatively and quantitatively using gas chromatography-mass spectrometry (GC-MS) and gas chromatography with flame ionization detection (GC-FID). We identified 16 constituents of the leaves of tree A, 18 in tree B, and 14 in tree C. The volatiles of the 3 trees differ in the respective amounts of monoterpenoids, hydrocarbons, fatty acids, and their methyl esters. The following constituents were identified in all of the 3 trees in largest percentage: hexahydrofarnesyl acetone (23.6%, 16.0%, and 27.7%), α-linolenic acid methyl ester (14.8%, 20.7%, and 15.1%), and pentacosane (22.2%, 22.4%, and 21.9%). Other identified compounds include the monoterpenes ( E)- α-ionone and ( E)- β-ionone.

A thin-layer chromatographic (TLC)-densitometric method was used for the parallel quantification of rosmarinic acid (RA) and caffeic acid (CA) in crude extracts of Salvia species (Family Lamiaceae), obtained by ultrasonic extraction with 60% methanol. The densitometric measurement was performed in fluorescent mode as it has been published earlier. The applicability of the method has been investigated mainly from the viewpoint of the starting material. Questions are discussed like, what kind of factors should be taken into account, if the drugs are to be characterized, and how the RA and CA contents of samples vary in the plants are discussed. The drugs (plant material) show great differences due to the time of harvest, to the organ composition of drugs, and to the extraction and storage conditions of the stock-solutions prepared from them. The importance of these parameters is illustrated on Salvia species native to Hungary.

The antiinflammatory activities of aqueous extracts prepared from the aerial parts of ten Hungarian Stachys species were investigated in vivo in the carrageenan-induced paw oedema test after intraperitoneal and oral administration to rats. Some of the extracts were found to display significant antiphlogistic effects when administered intraperitoneally and orally; in particular, the extracts of S. alpina, S. germanica, S. officinalis and S. recta demonstrated high activity following intraperitoneal administration. At the same dose of 5.0 mg/kg, these extracts exhibited similar or greater potency than that of the positive control diclofenac-Na. The main iridoids present in the investigated extracts, ajugoside, aucubin, acetylharpagide, harpagide and harpagoside, were also assayed in the same test, and high dose-dependent antiphlogistic effects were recorded for aucubin and harpagoside. These results led to the conclusion that most probably iridoids are responsible for the antiinflammatory effect of Stachys species, but other active constituents or their synergism must also be implicated in the antiinflammatory effect.

Members of the Salvia genus, like many representatives of the subfamilies Lamioideae and Nepetoideae, can be grown under the continental conditions of Hungary, beyond their native area of distribution. Salvia tomentosa Mill., S. officinalis L., S.fruticosa Mill., S. lavandulifolia Vahl., S. candelabrum Boiss., and S. ringens Sm. are species with essential oils of similar characteristics, i.e., with a predominant monoterpenoid content, whereas the thujone shows significant variations. The investigations reviewed have demonstrated that since S. tomentosa Mill. has a similar essential oil composition to S. officinalis L., but with a favorably lower thujone content, it could be regarded as a potential substitute for the purpose of essential oil production. Among the non-volatile components, ursolic, oleanolic, rosmarinic, caffeic acids do not show remarkable differences in the species investigated.

Different parts of three Ballota species ( B. nigra, B. hirsuta , and B. rupestris ) have been analyzed for phenylpropanoid content by means of a simple extraction technique and a newly established TLC-densitometric method. The compounds studied were verbascoside (VE), forsythoside B (FB), and caffeoyl-malic acid (CM). The largest amounts were detected in B. rupestris - CM (0.30%) and VE (4.58%) in the leaves and FB (3.0%) in the roots, followed by B. nigra ; the smallest amounts were found in B. hirsuta - 0.08% CM, 1.64% VE, and 0.91% FB in the leaves. The absence of caffeoylmalic acid was characteristic of the roots only. B. nigra , the common representative of the genus in Hungary, was chosen for thorough study of variation of phenylpropanoids during the vegetative period. Amounts of the phenylpropanoids increased during the main and secondary flowering periods in June (FB, 6.00%; VE, 8.50%; CM, 0.45% in the leaves) and September (FB, 3.80%; VE, 7.00%;, CM, 0.35% in the leaves).

Aqueous and organic extracts of 25 selected species from four tribes of Hungarian Asteraceae were screened in vitro for antiproliferative activity against HeLa (cervix epithelial adenocarcinoma), A431 (skin epidermoid carcinoma) and MCF7 (breast epithelial adenocarcinoma) cells, using the MTT assay. Twenty five of the 228 tested extracts from different parts of the species of Astereae (6), Inuleae (3), Heliantheae (5) and Anthemideae (11) demonstrated a substantial antiproliferative effect (at least 50% inhibition of cell proliferation) at 10 microg/mL against one or more of the cell lines. Complete dose-response curves were generated and IC(50) values were calculated for these active extracts, and their direct cytotoxic effects were determined. In summary, 11 of the tested 25 plants were found to be active and 4 of them (Anthemis ruthenica, Erigeron canadensis, Erigeron annuus and Inula ensifolia) had not been studied previously for either active compounds or anticancer properties.

We have isolated five iridoids (harpagide, acetylharpagide, harpagoside, ajugoside, and aucubin) from St. recta L. by a combination of chromatographic methods-NP-TLC, RP-HPLC, and TLC-densitometry. The isolated iridoids were identified on the basis of their physical and spectroscopic properties. Basic information about their structures was obtained from their NMR spectra. X-ray fluorescence spectroscopy can be used for direct determination of inorganic elements in the leaves, stems, and inflorescences of Stachys recta L. We identified K, Ca, Mo, Fe, Co, Ir, Ga, Tl, and S.

In the production and utilization of Medicinal and aromatic plants quality assurance has always been a focal issue. Despite this, the last decades have seen a new upsurge in the need to improve the traceability and safety of natural products. The increasing reliability in the production/collection practices of these species is also meant to contribute to the increasing acceptance of these commodities. To date, the major scientific bodies/organizations involved are engaged in the elaboration of guidelines and regulations that should contribute to the availability of validated scientific data on quality, safety and efficacy. Appropriate policies and legal frameworks to guide the protection, trade, and applications of medicinal and aromatic plant materials are also being elaborated (GAP, GCP, GMP, etc.). Due to the versatility, and the occasionally national character of quality assurance related activity there seems to be an increased need for the international harmonization of efforts.

Most of the beneficial biological effects (e.g. neurosedative, antioxidant, and antimicrobial) of black horehound ( Ballota nigra L.) are because of its phenylpropanoid glycoside content. A rapid TLC-densitometric method with acceptable accuracy has been established for quantitative analysis of the most characteristic components of black horehound. Experiments were performed to establish the optimum conditions for densitometric measurement and for storage of extracts. Practical application of the elaborated method is reported for five plant extracts.

Species of the Lamioideae subfamily of the family Lamiaceae , including Stachys species, do not contain much essential oil. The first stage of studying the volatile components of Stachys species was characterization of n -hexane extracts by TLC, with sabinene, limonene, linalool, and β-caryophyllene as test materials. On the basis of these results we then studied the volatile compounds by use of GC-FID and GC-MS. By examination of the TLC plates, use of Kovats retention indices, and MS library search we identified components of essential oil fractions obtained from Stachys officinalis II. (L.) Trev., Stachys alpina L., Stachys macrantha (Koch) Stearn, Stachys palustris L., Stachys recta L. subsp. recta, and Stachys annua L. (from Vacratot) by vapor distillation in accordance with Hungarian Pharmacopoeia VII.

From the n-hexane fraction of the fruits of Vitex agnus-castus, two labdane-type diterpenes, vitetrifolin B and C, were isolated by means of multiple chromatographic separations, together with the previously identified rotundifuran, vitexilactone and the sesquiterpene spathulenol. From the EtOAc fraction, eupatorin was identified for the first time, besides the known casticin, penduletin, vitexin and orientin. The n-hexane, EtOAc and MeOH-H2O fractions of the MeOH extract of Agni-casti fructus were subjected to in vitro antioxidant assays. The EtOAc extract displayed a significant concentration-dependent effect when tested by 1,1-diphenyl-2-picrylhydrasyl (DPPH) free radical assay (IC50 = 68 mu g/mL) and against the autooxidation of a standard rat brain homogenate (IC50 = 14 mu g/mL). The MeOH-H2O fraction was less active with 3643 mu g/mL (DPPH test) and IC50 = 125 mu g/mL (rat brain homogenate), while the n-hexane phase proved to be inactive. The main flavonoid constituents of the EtOAc extract, casticin, vitexin and orientin were assayed for antioxidant activity and found that only casticin possesses a marked lipid peroxidation inhibitory effect (IC50 = 0.049 mm) compared with that of the positive control ascorbic acid (IC50 = 0.703 mm). Copyright (c) 2007 John Wiley & Sons, Ltd.

The volatile constituents of both dried and fresh leaves, twigs and flowers of Philadelphus coronarius L. (Hydrangeaceae) were isolated by hydrodistillation and analyzed by GC/FID and GC/MS. There were major differences between the various oils. For example, epi-13-manool (47.6 %) was the major component of the fresh flower oil, but was not detected in the dried fower oil, whereas isolongifolol (15.5%), 2-nonanol (11.1%) and 7-hydroxycoumarin (9.7%) were the principal components of the dried flower oil. In the oils of both the fresh and dried leaves the major constituent was (E,E)-farnesol, although the proportion of this compound in the fresh leaf oil was considerably higher (37.3%) than that of the dried leaf oil (3.2%). (E,E)-farnesol was also the major component of the fresh twig oil (16.6%), but was absent from the dried twig oil, the major constituent of which was (E,E)-2,4-decadienal (2.4%); however, this was not detected in the fresh twig oil.

Stachys species contain iridoids, for example harpagide, acetylharpagide, aucubin, harpagoside, and ajugoside. The iridoid composition and content of leaves, stems, and inflorescences of ten Stachys species have been compared by use of a TLC-densitometric method. Harpagoside, harpagide, and acetylharpagide proved to be the main iridoids in Stachys officinalis L., St. sylvatica L., St. grandiflora Host., St. macrantha (C. Koch.) Jalas I., St. alpina L., St. palustris L., St. recta L., St. byzantina Koch., and St. germanica L. No iridoids were detected in St. annua L.

The oleanolic and ursolic acid contents from 88 taxa of Lamiaceae (19 genera, 66 species, 8 subspecies, 9 varieties and 5 hybrids) were investigated using gas chromatography. Both triterpenoid acids were present in all of the investigated taxa, but the plants belonging in the subfamily Nepetoideae produced significantly higher amounts than those in the subfamily Lamioideae. The oleanolic acid content ranged from traces to 1.840% dry weight, and that of ursolic acid from traces to 4.019% dry weight.

Stachys palustris L. (Lamiaceae) contains the iridoids harpagide and acetylharpagide. The presence of other components, for example aucubin, can be revealed by TLC and RP HPLC. A readily applicable simple TLC-densitometric method for analysis of aqueous extracts of the plants has been established for comparative studies of aucubin-like iridoids in Stachys palustris.

Eleven mobile phases and three stationary phases have been investigated for TLC separation of a large number of ecdysteroids. Optimization of the chromatography was based on trial and error. Use of four mobile phases on three stationary phases enabled separation of all the ecdysteroids from each other in at least one system. The TLC behavior of ecdysteroids containing different numbers of hydroxyl groups, side-chain variations, and extra double bonds, and of positional isomers and stereoisomers, is reported and interpreted. Our method is suggested for screening plant extracts for ecdysteroids.

Acovulparine (=1α,6β,7R,8β,14α,16β)-4-(hydroxymethyl)-1,6,14,16-tetramethoxyaconit-19-ene-7,8-diol; 1), a new norditerpene alkaloid, was isolated from the MeOH extract of the whole plants of Aconitum vulparia Reichenb., together with the known compounds lycoctonine (2) and delcosine (3). The structures were established by HR-EI-MS and advanced 2D-NMR, including 1H-NMR, JMOD, 1H,1H-COSY, HSQC, and HMBC experiments. Acovulparine was found to contain the rare C(19)N azomethine group.

A new natural ecdysteroid, 9beta,20-dihydroxyecdysone (1) and four related compounds 5alpha-20-hydroxyecdysone (2), 5alpha-2-deoxy-integristerone A (3), integristerone A (4) and 22-deoxy-integristerone A (5) were isolated from the herb of Silene italica ssp. nemoralis. Compound 1 is the C-9 epimer of the known 9alpha,20-dihydroxyecdysone (6) and represents a peculiar steroid skeleton. The structures of the compounds were elucidated by 1D and 2D NMR, IR and MS spectroscopy.

A suitable combination of preparative scale separation methods results in effective clean-up of the ecdysteroids of Silene italica ssp. nemoralis (Waldst. and Kit.) Nyman. The isolation of minor ecdysteroids from the partially purified extract is based on the use of both droplet counter-current chromatography and low-pressure reversed-phase liquid chromatography. The purification is completed by preparative thin-layer chromatography and preparative high-performance liquid chromatography to obtain the minor ecdysteroids, such as 2-deoxy-20-hydroxyecdysone, shidasterone, 2-deoxy-polypodine B, makisterone C, and 9alpha,20-dihydroxyecdysone.

A methanolic extract of the aerial parts of Salvia candelabrum was subjected to multiple chromatographic separation under the guidance of anti-lipid peroxidation assay. From the most active fractions seven abietane and seco-abietane diterpenes were isolated by preparative TLC purification. Besides candesalvoquinone, candelabroquinone, 12- O-methylcandesalvone B, candesalvone B methyl ester and candelabrone (all reported earlier), the known candesalvone B and the new candesalvolactone were identified. The structures were established by means of mass spectroscopy and advanced 2D NMR methods. All the identified compounds were evaluated for antioxidant activity in enzyme-dependent (IC (50) values 3.49 - 10.42 microM) and enzyme-independent (IC (50) values 1.40 - 13.40 microM) systems of lipid peroxidation. All compounds displayed marked concentration-dependent effects in both tests as compared with those of authentic ascorbic, rosmarinic and caffeic acids. The differences in antioxidant capacities observed in the enzyme-independent system allowed conclusions concerning structure-activity relationships.

Five new diterpene polyesters, 1–5, with jatrophane skeletons were isolated from the fresh whole plants of Euphorbia serrulata. The structure elucidation was performed by means of UV/VIS spectroscopy, HR-ESI-MS, and advanced two-dimensional NMR methods, including 1H-NMR, JMOD, 1H,1H-COSY, NOESY, HMQC, and HMBC experiments. The relative configurations of 1–5 and their conformations in solution were analyzed on the basis of NOESY measurements. As a result of detailed NMR studies, complete 1H and 13C chemical-shift assignments of the compounds were possible. The isolated compounds differ stereochemically and do not comprise a uniform series regarding the configurations at C(2), C(6), and C(13). Compound 5 possesses the new structural feature of a double bond with (Z)-configuration in the macrocyclic ring of the jatrophane skeleton, while compound 2 has a CC bond in the five-membered ring, this being the first observation of this structural feature in the type of macrocyclic Euphorbiaceae diterpenes.

The volatile oil compositions of five Stachys species (Stachys officinalis L., St. grandiflora Host., St. byzantina Koch., St. germanica L. and St. sylvatica L.) have been studied using a GC/MS technique. All species gave low yields of oil. From St. officinalis 38 compounds were separated and partly identified, from St. grandiflora 24, St. byzantina 30, St. germanica 29 and St. sylvatica 43. All the samples contained o-ocymene, β-phellandrene, γ-terpinene, linalool, transand cis-pinocamphone, βcaryophyllene, δ-cadinene and α-cadinol. The steam distilled oils were poor in monobut rich in sesquiterpenes. Some of the sesquiterpenes, like germacrene D, epi-cubebol, and αand γ-muurolene are reported in Stachys species for the first time. The iridoids, harpagide and 8-O-acetyl-harpagide, were identified by TLC in the MeOH extracts of St. sylvatica, St. byzantina, St. officinalis (Betonica officinalis) and Betonica serotina. Taking other chemical constituents also into account Stachys species seem to be similar to those of the subfam. Lamioideae (according to Erdtman’s classification). INTRODUCTION In the course of our systematic chemotaxonomic investigation of the family Lamiaceae (Mathe et al., 1993), we have found the genus Stachys especially interesting for detailed study not only as it is the second largest genus of the family, but also as its chemistry is little known. There exist some uncertainties both in the place of the genus in the family and in the distinction between the genera Stachys and Betonica. The family Lamiaceae has been classified in different ways by Briquet, Bentham, Erdtman, and Cantino and Sanders (Cantino and Sanders, 1986) and, more recently by Cantino, Harley and Wagstaff (1992). Unlike Erdtman’s system, Bentham and Briquet, in their eight subfamilial (Bentham named ‘tribes’) system, regard the genus Stachys as a member of the subfam. Stachyoideae (tribe Stachydeae in Bentham’s system), which has morphological features characteristic of the subfam. Lamioideae in Erdtman's two subfamilial system. On the basis of literature, Stachys species contain iridoids and caffeic acid, lack rosmarinic acid, but yield essential oils. So, from chemical viewpoints, this genus may be a member of Erdtman's subfam. Lamioideae (Cantino and Sanders, 1986, Mathe, et al., 1993, Hegnauer, 1996,1990). The genus also shows some characteristics with regard to flavonoid contents (Kotsos, 2000; Skaltsa, 2000; Barberan, 1988; Skaltsa et al., 1999; Adams 1995). To verify some of these points, five Stachys species, either native to or easily cultivated in Hungary, have been studied. These are Stachys officinalis (L.) Trevis. (Betonica officinalis L.), St. grandiflora Host., St. byzantina Koch., St. germanica L. and St. sylvatica L. The rightness of the segregation of Stachys officinalis, as Betonica officinalis, from the genus Stachys is also discussed. MATERIALS AND METHODS Plants were cultivated in the experimental field of the Research Institute of Ecology and Botany of the Hungarian Academy of Sciences (at Vacratot, 40 km north of

Exploring new biologically active ingredients of plants are in the focus of medicinal plant researches. The chemicals of plants determine the healing properties of plants. This chapter deals with the active ingredients that can explain the applicability of medicinal plants or can help in the evaluation of the plants from chemical point of view. Rather difficult thing to classify the medicinal plants on the bases of their active chemical ingredients as in the same plants several compounds belonging to various chemical groups can be found and, in addition, in many cases they contribute to the effect and applicability of the plants. In other cases the same type of compounds is present in plants bearing different utilization from medical viewpoint. Exceptions are the plants having compounds of strong biological effect and the ingredients from them as pure compounds are used after their isolation. In this chapter the plants are discussed according to the classical, broadly applied grouping: based upon the biosynthetic pathways of the main active ingredients. Consequently, the classes of primary and secondary metabolites are used. Primary metabolites are as follows: carbohydrates, fatty acids, fats, proteins. The main groups of secondary metabolites discussed are the phenolics (including polyketides), terpenoids and alkaloids. Examples, most frequently occurring in medicinal plant handbooks illustrate the chemical diversity of these compounds.

Seven alkaloids were isolated from Sprekelia formosissima, and five from Hymenocallis x festalis. Tazettine, lycorine, haemanthidine and haemanthamine were evaluated for antiproliferative and multidrug resistance (mdr) reversing activity on mouse lymphoma cells. Lycorine, haemanthidine and haemanthamine displayed pronounced cell growth inhibitory activities against both drug-sensitive and drug-resistant cell lines, but did not significantly inhibit mdr-1 p-glycoprotein. Thus, the tested alkaloids are apparently not substrates for the mdr efflux pump. Assays for interactions with DNA and RNA revealed that the antiproliferative effects of lycorine and haemanthamine result from their complex formation with RNA.

5-Alpha-2-deoxyintegristerone A and 5-beta-2-deoxyintegristerone A were isolated from the aerial parts of Silene italica ssp. nemoralis (Waldst. and Kit.) Nyman using a specific combination of absorption column chromatography, preparative thin-layer chromatography and preparative HPLC. Both normal-phase and reversed-phase modes of HPLC were employed for isolation. Structural elucidation of 5-alpha-2-deoxyintegristerone A was completed by X-ray diffraction. Both 5-alpha-2-deoxyintegristerone A and 5-beta-2-deoxyintegristerone A were firstly isolated from this plant. We propose that 5-alpha-2-deoxyintegristerone A is not an artifact but an integral part of the ecdysteroid spectrum of Silene italica ssp. nemoralis (Waldst. and Kit.) Nyman.

The extract of Artemisia asiatica herb with antiproliferative activity against four human tumor cell lines (A2780, A431, HeLa, and MCF7) was analyzed by the MTT assay, and bioassay-directed fractionation was carried out in order to identify the compounds responsible for the cytotoxic activity. Guaianolide (1–4), seco-guianolide (5), germacranolide (6) and eudesmanolide sesquiterpenes (7), monoterpenes (8, 9), including the new compound artemisia alcohol glucoside (8), and flavonoids (10–16) were isolated as a result of a multistep chromatographic procedure (CC, CPC, PLC, and gel filtration). The compounds were identified by means of UV, MS, and NMR spectroscopy, including 1H-and 13C-NMR, 1H-1H COSY, NOESY, HSQC, and HMBC experiments. The isolated compounds 1–16 were evaluated for their tumor cell growth-inhibitory activities on a panel of four adherent cancer cell lines, and different types of secondary metabolites were found to be responsible for the cytotoxic effects of the extract. Especially cirsilineol (13), 3β-chloro-4α,10α-dihydroxy-1α,2α-epoxy-5α,7αH-guai-11(13)-en-12,6α-olide (3), and iso-seco-tanapartholide 3-O-methyl ester (5) exerted marked cytotoxic effects against the investigated cell lines, while jaceosidin (12), 6-methoxytricin (15), artecanin (2), and 5,7,4′,5′-tetrahydroxy-6,3′-dimethoxyflavone (14) were moderately active. All the sesquiterpenes and monoterpenes are reported here for the first time from this species, and in the case of artecanin (2), 3α-chloro-4β,10α-dihydroxy-1β,2β-epoxy-5α,7αH-guai-11(13)-en-12,6α-olide (4), ridentin (6), and ridentin B (7), previously unreported NMR spectroscopic data were determined.

Seasonal dependence of 20-hydroxyecdysone content of Serratula wolffii was monitored. Thin-layer chromatography was used after a single clean-up using polyamide. High performance liquid chromatography was employed for the analysis, after removal of the high excess of flavonoids and other impurities using a clean-up on both C18 and polyamide. Parallel analyses using both thin layer chromatography with densitometry and high performance liquid chromatography gave similar results at the determination of vegetation dependence of ecdysteroids. Results indicated two maxima of ecdysteroid content (w/w), such as beginning of vegetation (in April and May) and during blossoming (in August). The two ecdysteroids, 20-hydroxyecdysone and polypodine B were also isolated from Serratula wolffii, their chemical structures were identified by spectroscopic methods.

5α-Dihydrorubrosterone (2β, 3β, 14α, 17β-tetrahydroxy-5α-androst-7-ene-6-one), a new 19-carbon 5α-ecdysteroid, was isolated together with its 5β counterpart from the aerial parts of Silene otites L. (Wib.) (Caryophyllaceae) by a combination of solvent partition, low-pressure column chromatography, thin-layer chromatography (normal-phase and reversed-phase) and finally HPLC. Mass spectrometry and nuclear magnetic resonance spectroscopic procedures were used for compound characterization. Copyright © 2000 John Wiley & Sons, Ltd.

From the acetone extract of the dry roots of Salvia glutinosa, 15 diterpenoids were isolated and characterised. Of these, 12-deoxydanshenxinkun B and dihydroisotanshinone II are novel compounds and isotanshinone II is a new natural product. The acetone extracts of S. austriaca, S. tomentosa and S. verticillata all yielded 7α-acetoxyroyleanone and 7α-hydroxyroyleanone. In addition, royleanone and 6,7-dehydroxyroyleanone were isolated from S. tomentosa.

The rosmarinic and caffeic acid contents of 96 Lamiaceae taxa (20 genera, 76 species, 8 subspecies, 9 varieties, 3 hybrids) were investigated using a TLC-densitometric method. Rosmarinic acid was present in all species of the subfamily Nepetoideae, but was absent from those in the subfamily Lamioideae. However, with the exception of 3 Lamioideae species, caffeic acid was found in measurable quantities in all taxa studied. The rosmarinic acid content ranged from 0.01 to 9.30 mg/g, while that of caffeic acid varied from 0 to 0.62 mg/g.

From the roots of Salvia nutans seven diterpenoids have been isolated and characterized, primarily from spectroscopic data. Three of the compounds are new 12-deoxyroyleanones (12-deoxy-6,7-dehydroroyleanone, 12-deoxy-6-hydroxy-6,7-dehydroroyleanone and 12-deoxy-7,7-dimethoxy-6-ketoroyleanone).

From the pro-inflammatory active extract of Euphorbia peplus, a new diterpene polyester (1) based on the jatrophane skeleton was isolated together with the known compounds 2-5. The irritant activities of some jatrophane diterpenes (2, 3 and 6-9) were also investigated: only compound 2 was found to exert a weak pro-inflammatory activity on mouse ear.

The cytotoxic activity of Stachys plants and of aucubin and harpagide against MCF7-breast adenocarcinoma, HeLa-cervix adenocarcinoma, A431-skin carcinoma of epithelial origin is reported in this study. Cisplatin and doxorubicin were use as reference compound.

Aerial parts of 23 species distributed in 10 genera of the Amaranthaceae have been examined for the presence of betaines. Glycinebetaine was isolated from all the species studied and, in addition, trigonelline was detected in eight out of the nine species of Amaranthus , 1 of the two species of Alternanthera and in the species of Iresine (3), Celosia (2), Chamissoa (1), Aerva (1), Gomphrena (1) and Froelichia (1). With the exception of I. herbstii , glycinebetaine was the predominant betaine. The highest yield of this compound was from Cyathula geniculata (2.11%, dry weight), but, with the exception of I. herbstii (0.05%), the species tested had contents in the range 0.28–2.11%, dry weight. Trigonelline yields varied from 0.004 to 0.15%, dry weight. From the data obtained, classification of the Amaranthaceae as a betaine – accumulating family would appear to be justified.

Several new minor ecdysteroids from Silene otites (Caryophyllaceae) have been purified and identified. This plant species had previously been shown to contain a complex ecdysteroid cocktail, with 20-hydroxyecdysone as the major component, and significant amounts of 2-deoxyecdysone, 2-deoxy-20-hydro-xyecdysone, and 20-hydroxyecdysone 22-acetate, and a set of minor ecdysteroids. The use of powerful techniques for purification and spectroscopic analyses has now allowed the isolation and identification of more than 30 different molecules including 21-hydroxylated ecdysteroids, thus adding a new position of the carbon skeleton that can be modified in ecdysteroids. Thus, in S. otites, a complex array of individual reactions can be used in various combinations leading both to major and minor components. Deciding whether this represents a random process without biological consequences or if (some of) the various minor components may play a specific role in, e.g., insect-plant relationships, will require the extensive use of appropriate in vitro and in vivo bioassays.

Two new jatrophane diterpenoids, esulatin D and E, have been isolated from the dichloromethane extract of the whole, undried plant of Euphorbia esula. The structures have been assigned on the basis of spectral analysis, including 2D NMR experiments.

Six minor new ecdysteroid components have been isolated from Silene otites (L.) Wib. by a combination of chromatographic methods. Three of them (2-deoxy-20-hydroxyecdysone 3,22-diacetate, 5 α -2-deoxy-20-hydroxyecdysone 3-acetate, and 2-deoxy-20-hydroxyecdysone 3-crotonate) are new natural products.

Three new jatrophane diterpenes, esulatin A, B and C (1-3) were isolated and characterized from the whole, undried plant of Euphorbia esula. By means of spectral analysis, the structures were established as penta- and heptaesters of hitherto unknown, polyfunctional diterpene parent alcohols. Esulatin A (1) and C (3) are the diterpenoids with the highest degree of esterification identified to date from the family Euphorbiaceae.

Essential oils of Haplophyllum tuberculatum (Forssk.) A. Juss., produced from plants collected in the United Arab Emirates, were analyzed by GC and GC/MS. The composition of the oils varied considerably. Those obtained from plants collected in May (1997 and 2001) were similar, with α-phellandrene (10.7–32.9%) being the major component and with signifcant amounts of β-caryophyllene (6.3–12.8%), β-pinene (7.6–8.0%), limonene (4.0–9.6%) and δ-3-carene (5.5–6.0%). However, the oil distilled from plants collected in April (1998) had major components of linalool (15.0%), linalyl acetate (10.6%), β-caryophyllene (9.7%) and α-terpineol (6.7%). The compositions of all three oils prepared from UAE collected plants differed signifcantly from those reported for the same species col-lected in Egypt and Iran.

The object of our work was the identification and quantification of inorganic elements in Ginkgo biloba L. leaves (Ginkgonis folium, Ginkgoaceae) by X-ray fluorescence analysis. The plant material was obtained from a 50-years-old female tree at the Comenius University Botanical Garden (Bratislava, Slovakia). Leaves were collected from early May to late September, with the last sample consisting of fallen leaves. The elements analyzed were: phosphorus, sulfur, potassium, calcium, scandium, iron, zinc, yttrium, molybdenum, tellurium, samarium, gadolinium, dysprosium, iridium, thallium and lead. The amounts of the monitored heavy metals were below the limits specified in Ph. Eur. 7 and PhS 1.