Gentiana Lutea Research Articles

Analysis of iridoids, secoiridoids and xanthones in Centaurium erythraea, Frasera caroliniensis and Gentiana lutea using LC–MS and RP-HPLC

This study presents a new and validated HPLC method for the simultaneous determination of bioactive compounds in Centaurium erythraea, Frasera caroliniensis and Gentiana lutea. The iridoid loganic acid, four secoiridoids and 29 xanthones were separated on a RP-18 column, using aqueous o-phosphoric acid (0.085%, v/v) and acetonitrile as mobile phase. Phytochemical investigation of C. erythraea herb and F. caroliniensis roots resulted into isolation of 25 xanthones and three secoiridoids the structure of which was elucidated by spectroscopic means (NMR, MS and UV). 1,3,8-Trihydroxy-5,6-dimethoxyxanthone, isolated from C. erythraea, turned out to be a novel xanthone. The stability of the analytes was tested by subjecting samples to light, moisture and different temperatures. After six months of storage, decomposition of gentiopicroside and sweroside was observed. The swertiamarin content was nearly unchanged when stored at room temperature or in the refrigerator, but high temperature conditions reduced the content to 85%. In contrast, xanthones were stable under long-term, refrigerated and accelerated conditions. The established chromatographic method has been successfully applied for the quantification of the bioactive compounds in the three plants. The presence and distribution of polyoxygenated xanthones within the three members of the Gentianaceae family and their significance as analytical markers are discussed.

Evaluation of the antioxidants activities of four Slovene medicinal plant species by traditional and novel biosensory assays

We investigated the antioxidant activity of methanolic and water extracts of Slovene accessions of four medicinal plant species ( Salvia officinalis, Achillea millefolium, Origanum vulgare subsp. vulgare and Gentiana lutea). Their free radical-scavenging activity against the DPPH· free radical was studied with a spectrophotometric assay, while their biological activity with the help of a laboratory-made biosensor based on immobilized fibroblast cells (assay duration: 3 min). The observed antioxidant activity of the extracts from the four investigated medicinal plant species was dependent on both the solvent used for extraction and the assay method (conventional or biosensor-based). Independently from the assay method and the solvent used for extraction, the lowest scavenging activity was observed in root extracts of G. lutea. Treatment of the immobilized cells with the plant extracts resulted in an increase of the cell membrane potential (membrane hyperpolarization), possibly due to the reduction of membrane damage due to oxidation. The novel cell biosensor could be utilized as a rapid, high throughput tool for screening the antioxidant properties of plant-derived compounds.

Low-dose exposure to Veratrum album in children causes mild effects – a case series

Introduction. White or false hellebore (Veratrum album) has a toxicological relevance because of the potential for misidentification of this plant as yellow gentian (Gentiana lutea). Case series. We report a retrospective case series of 11 children (8–12 years) with accidental intake of V. album at a youth camp where they had collected herbs for preparing fresh herb tea. Two children (18%) remained asymptomatic. Nine (82%) developed mild gastrointestinal symptoms, six (55%) presented neurological symptoms, and three (27%) showed bradycardia. All children recovered completely within 10 h of ingestion. The plant was identified at the emergency department; however, detection of veratridine and cevadine by means of high-performance liquid chromatography–Mass spectrometry from the blood of the child with the most severe symptoms was negative (limit 0.01 ng/mL). Discussion. Veratrum species contain more than 200 different alkaloids, which are the principal toxins and are responsible for most clinical symptoms. There are likely multiple mechanisms of toxicity and some of them are only partially understood. The opening of voltage-gated sodium channels is probably one of the most relevant pathophysiological mechanisms. Conclusions. Veratrum album intoxication in children demonstrated the same clinical course as observed in adults. Accidental ingestion of a low dose of the plant had a favorable outcome with supportive care.

Antiviral activity of a composition of Gentiana lutea L., Primula veris L., Sambucus nigra L., Rumex spec. andVerbena officinalis L. (Sinupret®) against viruses causing respiratory infections

Sinupret ® , an aqueous-ethanolic extract from five plants, Gentiana lutea L., Primula veris L., Sambucus nigra L., Rumex spec. and Verbena officinalis L., is frequently used in the treatment of acute and chronic rhinosinusitis and respiratory viral infections such as common cold. To date little is known about basic mechanisms of its potential antiviral activity. Therefore experiments have been performed for the detection of its antiviral activity against a broad panel of human pathogenic enveloped and non-enveloped RNA and DNA viruses causing infections of the upper respiratory tract: influenza A virus, parainfluenza virus, human rhinovirus B, coxsackievirus, adenovirus C, and respiratory syncytial virus. Determination of virus production was performed after treatment of the infected cells with non-toxic concentrations of the commercially available product using plaque-reduction assays, analyses of cytopathogenic effects and ELISAs for viral proteins. Antiviral activity of Sinupret ® could be detected independent of the type of the viruses in RNA as well as in DNA virus infected cell cultures and also against coated and uncoated viruses. A very strong inhibitory activity was obvious against adenovirus and respiratory syncytial virus infections. These results demonstrate that Sinupret ® showed a broad antiviral activity which basic mechanisms are still unclear. This has to be elucidated in further studies.

Studies concerning the phenotipical variability of some Gentiana lutea L. genotipes.

The ever increasing demands for raw materials obtained from Gentiana lutea L. species. will lead to introducing this species into culture. The cultivators will have the chance to obtain significant earnings by selling the production. For this. we identified 10 families that can be a prime source of germplasm for some amelioration. They were subjected to an analysis study of the variability of diverse quantitative elements. In the present article. we show the correlations carried out between different characters: the parcel diameter. the length of the rhizome. the number of ramifications. total mass of subterranean organs. height. number of knots. number of bastes. total number of fruits on the plant. The superior quality of the raw material obtained trough the cultivation methods established as the research we developed and extending a superior cultivator into the culture. will decrease the temptation of harvesting our country’s spontaneous flora. this species having a small habitat and a small density. that do not allow satisfying the industry’s demand.

Accidental Intoxication with Veratrum album

A 49-year-old man consumed two glasses (approximately 2 x 20 mL) of a beverage containing yellow gentian (Gentiana lutea). Shortly after ingestion, he developed nausea, vomiting, and oral paraesthesia. On admission to the hospital he suffered from severe bradycardia (35 beats/min) and hypotension (50/30 mm Hg), and he was treated with activated charcoal, antiemetics (metoclopramide, ondansetron), atropine, and intravenous electrolytic solution. The initial suspicion of Veratrum poisoning could be confirmed by identifying protoveratrines A (ProA) and protoveratrine B (ProB) in a sample from the beverage as well as in the patients serum by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS). The yellow-colored beverage contained 25% ethanol (by headspace gas chromatography), 20.4 mg/L ProA, and 13.7 mg/L ProB. The serum concentration of ProA was 1162 ng/L and ProB was 402 ng/L. Veratridine, cevadine, and jervine were not detected, neither in the beverage nor in the serum sample. The lower limits of quantitation for all compounds is 10 microg/L (S/N > 10, beverage) and 100 ng/L (S/N > 10, serum). After treatment, the patient completely recovered from the symptoms within 24 h and was discharged from the hospital. The analytical method described was developed for the simultaneous identification and quantitation of five Veratrum alkaloids. The method is based on a liquid-liquid extraction followed by LC-MS-MS analysis. The time needed for analysis was 6 min.

Functional analysis of multiple carotenogenic genes from Lycium barbarum and Gentiana lutea L. for their effects on β-carotene production in transgenic tobacco

Carotenoids are red, yellow and orange pigments, which are widely distributed in nature and are especially abundant in yellow-orange fruits and vegetables and dark green leafy vegetables. Carotenoids are essential for photosynthesis and photoprotection in plant life and also have different beneficial effects in humans and animals (van den Berg et al. 2000). For example, beta-carotene plays an essential role as the main dietary source of vitamin A. To obtain further insight into beta-carotene biosynthesis in two important economic plant species, Lycium barbarum and Gentiana lutea L., and to investigate and prioritize potential genetic engineering targets in the pathway, the effects of five carotenogenic genes from these two species, encoding proteins including geranylgeranyl diphosphate synthase, phytoene synthase and delta-carotene desaturase gene, lycopene beta-cyclase, lycopene epsilon-cyclase were functionally analyzed in transgenic tobacco (Nicotiana tabacum) plants. All transgenic tobacco plants constitutively expressing these genes showed enhanced beta-carotene contents in their leaves and flowers to different extents. The addictive effects of co-ordinate expression of double transgenes have also been investigated.

Genotype and plant growth regulator-dependent response of somatic embryogenesis from Gentiana spp. leaf explants

Gentiana kurroo (Royle), Gentiana cruciata (L.), Gentiana tibetica (King. ex Hook. f.), Gentiana lutea (L.), and Gentiana pannonica (Scop.) leaves derived from axenic shoot culture were used as explants. For culture initiation, leaves from the first and second whorls from the apical dome were dissected and cultured on Murashige and Skoog (MS) basal medium supplemented with three different auxins: 2,4-dichlorophenoxyacetic acid, 1-naphthaleneacetic acid (NAA), or 3,6-dichloro-o-anisic acid (dicamba) in concentrations of 0.5, 1.0, or 2.0 mg/l; and five different cytokinins: zeatin, 6-furfurylamonopurine (kinetin), N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ), N-(2-chloro-4-pyridyl)N′-phenylurea, or 6-benzylaminopurine (BAP). The cytokinin concentrations used were dependent on the type of cytokinin and varied between 0.25 and 3.0 mg/l. After 2 mo. of culture, the morphogenic response of explants was assessed. Frequency of embryogenesis was the highest for G. kurroo (54.7%) and dependent on plant growth hormones (PGRs). This gentian was the only species showing morphogenic capabilities on media supplemented with all applied combinations of PGRs, while none of the 189 induction media permutations stimulated somatic embryogenesis from G. lutea explants. G. tibetica and G. cruciata both produced an average of 6.6 somatic embryos per explant, while G. pannonica and G. kurroo regenerated at 15.7 and 14.2 somatic embryos per explant, respectively. Optimum regeneration was achieved in the presence of NAA combined with BAP or TDZ. This auxin also stimulated abundant rhizogenesis. Somatic embryos were also regenerated from adventitious roots of G. kurroo, G. cruciata, and G. pannonica. Somatic embryos converted into plantlets on half strength MS medium.

Down-regulation of NR2B receptors partially contributes to analgesic effects of Gentiopicroside in persistent inflammatory pain

Gentiopicroside is one of the secoiridoid compound isolated from Gentiana lutea. It exhibits analgesic activities in the mice. The anterior cingulate cortex (ACC) is a forebrain structure known for its roles in pain transmission and modulation. Painful stimuli potentiate the prefrontal synaptic transmission and induce glutamate NMDA NR2B receptor expression in the ACC. But little is known about Gentiopicroside on the persistent inflammatory pain and chronic pain-induced synaptic transmission changes in the ACC. The present study was undertaken to investigate its analgesic activities and central synaptic modulation to the peripheral painful inflammation. Gentiopicroside produced significant analgesic effects against persistent inflammatory pain stimuli in mice. Systemic administration of Gentiopicroside significantly reversed NR2B over-expression during the chronic phases of persistent inflammation caused by hind-paw administration of complete Freunds adjuvant (CFA) in mice. Whole-cell patch clamp recordings revealed that Gentiopicroside significantly reduced NR2B receptors mediated postsynaptic currents in the ACC. Our findings provide strong evidence that analgesic effects of Gentiopicroside involve down-regulation of NR2B receptors in the ACC to persistent inflammatory pain.

Ketoconazole-induced testicular damage in rats reduced by Gentiana extract

Ketoconazole (KET) is an antifungal drug with a broad spectrum of activity that also induces reproductive toxicity in humans and animals. The protective effect of Gentiana (GEN) extract ( Gentiana lutea) against KET-induced testicular damage was evaluated in male Wistar rats. GEN extract was administered orally (1 g/kg b wt/day) for 26 days. Three weeks after extract administration, KET was co-administered intraperitoneally at a dose of 100 mg/kg once a day for 5 days. KET-induced reproductive toxicity was associated with clear reductions of the weights of testes and epididymides, sperm indices and serum testosterone levels. KET also induced severe testicular histopathological lesions such as degeneration of the seminiferous tubules and depletion of germ cells. In addition, marked oxidative damage to testicular lipids and alterations of natural antioxidants (catalase (CAT) and superoxide dismutase (SOD)) were reported in association with KET toxicity. Most of the KET-induced effects were greatly decreased with the concomitant application of GEN extract. This study suggests a protective role of GEN extract that could be attributed to its antioxidant properties.

Quantitative analysis of iridoids, secoiridoids, xanthones and xanthone glycosides in Gentiana lutea L. roots by RP-HPLC and LC–MS

The here described HPLC-method enables the determination of all major, currently known bioactive compounds in gentian roots. A separation of iridoids (loganic acid), secoiridoids (swertiamarin, gentiopicroside, amarogentin, sweroside), xanthones (gentisin, isogentisin) and two xanthone glycosides (gentiosides) was possible on RP-18 column material, using 0.025% aqueous TFA, acetonitrile and n-propanol as mobile phase. The method is sensitive (LOD ≤ 37 ng/ml and LOQ ≤ 112 ng/ml), accurate (recovery rates of spiked samples were between 96.7 and 101.5%), repeatable ( σ rel ≤ 1.7%) and precise (intra-day variation ≤ 4.6%, inter-day variation ≤ 3.1%). LC–MS experiments performed in negative ESI mode assured peak purity and identity. Analysis of several commercially available G. lutea samples showed that gentiopicroside is the most dominant compound in the specimens (4.46–9.53%), followed by loganic acid (0.10–0.76%), swertiamarin (0.21–0.45%) and the xanthone glycosides. Gentisin and isogentisin were found in much lower concentrations between 0.02 and 0.11%, respectively.

Screening of plant extracts for antimicrobial activity against bacteria and yeasts with dermatological relevance

There is cumulative resistance against antibiotics of many bacteria. Therefore, the development of new antiseptics and antimicrobial agents for the treatment of skin infections is of increasing interest. We have screened six plant extracts and isolated compounds for antimicrobial effects on bacteria and yeasts with dermatological relevance. The following plant extracts have been tested: Gentiana lutea, Harpagophytum procumbens, Boswellia serrata (dry extracts), Usnea barbata, Rosmarinus officinalis and Salvia officinalis (supercritical carbon dioxide [CO2] extracts). Additionally, the following characteristic plant substances were tested: usnic acid, carnosol, carnosic acid, ursolic acid, oleanolic acid, harpagoside, boswellic acid and gentiopicroside. The extracts and compounds were tested against 29 aerobic and anaerobic bacteria and yeasts in the agar dilution test. U. barbata-extract and usnic acid were the most active compounds, especially in anaerobic bacteria. Usnea CO2-extract effectively inhibited the growth of several Gram-positive bacteria like Staphylococcus aureus (including methicillin-resistant strains - MRSA), Propionibacterium acnes and Corynebacterium species. Growth of the dimorphic yeast Malassezia furfur was also inhibited by Usnea-extract. Besides the Usnea-extract, Rosmarinus-, Salvia-, Boswellia- and Harpagophytum-extracts proved to be effective against a panel of bacteria. It is concluded that due to their antimicrobial effects some of the plant extracts may be used for the topical treatment of skin disorders like acne vulgaris and seborrhoic eczema.

STUDIES CONCERNING THE PHENOTIPICAL VARIABILITY OF SOME GENTIANA LUTEA L. GENOTYPES

The ever increasing demands for raw materials obtained from Gentiana lutea L. species, will lead to introducing this species into culture. The cultivators will have the chance to obtain significant earnings by selling the production. For this, we identified 10 families that can be a prime source of germplasm for some amelioration. They were subjected to an analysis study of the variability of diverse quantitative elements. In the present article, we show the correlations carried out between different characters: the parcel diameter, the length of the rhizome, the number of ramifications, total mass of subterranean organs, height, number of knots, number of bastes, total number of fruits on the plant. The superior quality of the raw material obtained trough the cultivation methods established as the research we developed and extending a superior cultivator into the culture, will decrease the temptation of harvesting our country’s spontaneous flora, this species having a small habitat and a small density, that do not allow satisfying the industry’s demand.

The amount of secondary metabolites in cultivated Gentiana lutea L

The amount of secondary metabolites in cultivated Gentiana lutea L

Isogentisin—A novel compound for the prevention of smoking-caused endothelial injury

The best strategy in the fight against tobacco-induced diseases is prevention. However, more than one billion people around the world are smokers. Most of these people will develop or already suffer from tobacco-induced diseases. In this project, we screened 22 natural alpine plant extracts for their potential to protect human vascular endothelial cells from cigarette smoke-induced cell damage. Extracts from Gentiana lutea (Yellow Gentian) proved to be effective, and were therefore subjected to bio-guided fractionation. Although our analyses suggest that G. lutea contains several active principles, fractions containing isogentisin (1,3-dihydroxy-7-methoxyxanthone), and pure isogentisin, were most effective. In experiments addressing the nature of the mechanism of protection, we were able to show that isogentisin does not directly interfere with cigarette smoke chemicals. Addition of isogentisin to the cells as long as 4.5 h after exposure to cigarette smoke chemicals protected endothelial cells from cell death. Finally, detailed analyses of intracellular oxidative stress and protein oxidation suggest that isogentisin promotes cell survival by activating cellular repair functions.

Safe for saplings; safe for seeds?

In wood pastures spatial associations of tree saplings with nurse structures such as unpalatable plants are generally explained as associational resistance; i. e. reduced herbivory by association with a defended neighbour. However, these associations may result from other underlying processes that occur at the seed stage. Here we tested whether the observed associations between Picea abies tree saplings and nurse structures could also be explained by higher soil seed densities under versus outside nurse shrubs, or by reduced seed removal inside versus outside nurse structures. Further we tested for differential effects of the main seed predator groups on seed removal and establishment, both in gaps and in dense vegetation. We sampled in total 640 soil cores from inside and outside canopies of eight Crataegus monogyna shrubs and compared densities of P. abies seeds. Seed removal was studied inside and outside the canopies of four different nurse structures: C. monogyna, Gentiana lutea, rocks and tree stumps. The effects of cattle, birds, rodents and insects on seed removal and seedling establishment were studied using selective exclosures both in gaps and in dense vegetation. Soil seed densities under and outside shrubs did not differ significantly. Seed removal was considerable (85%), indicating the importance of seed removal as limiting factor for tree regeneration in wood pastures. In contrast to our expectations, seed removal was significantly higher under all four nurse structures then outside. The subsequent exclusion of cattle, birds and rodents reduced seed removal and increased establishment, but the individual contribution of each of these groups was relatively small, while the contribution of insects was greater. Seed removal was higher and, consequently, seedling establishment lower in dense vegetation than in the gaps. Our findings show that the earlier observed spatial associations between tree saplings and unpalatable plants, rocks and tree stumps are unlikely to be caused by an initially higher soil seed density or by reduced seed removal under these structures. Those structures were reported earlier to enhance tree sapling survival by offering protection against cattle grazing, but they appear not to protect tree seeds. This study shows the complexity of tree establishment in wood pasture ecosystems: apparently, safe sites for tree saplings are not safe for seeds.

Free radical scavenging activities of yellow gentian (Gentiana lutea L.) measured by electron spin resonance

Yellow gentian (Gentiana lutea L.) is a herbal species with a long-term use in traditional medicine due to its digestive and stomachic properties. This paper presents an investigation of the free radical scavenging activity of methanolic extracts of yellow gentian leaves and roots in two different systems using electron spin resonance (ESR) spectrometry. Assays were based on the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the superoxide radicals (O2*-) generated by the xanthine/xanthine oxidase (X/XO) system. The results of gentian methanolic extracts were compared with the antioxidant capacity of synthetic antioxidant butylated hydroxyanisole (BHA). This study proves that yellow gentian leaves and roots exhibit considerable antioxidant properties, expressed either by their capability to scavenge DPPH or superoxide radicals.

Seed bank persistence of clonal weeds in contrasting habitats: implications for control

The ability of weeds to form a seed bank is important for their population dynamics and management because it provides a refuge enabling reinvasion after established target plants have died. However, knowledge of the differential seed behaviour of individual species over multiple years and varying environmental conditions is surprisingly rare but necessary for effective control of diverse weed populations. We established a seed burial experiment in alpine habitats differing in management regime (i.e., forest, hay meadow and pasture) to determine whether seeds of the unpalatable perennial weeds, Veratrum album (white hellebore) and Gentiana lutea (yellow gentian) were able to delay germination and remain viable over 3 years. Our study shows that both species formed a short-term persistent seed bank; in the third-year, the soil seed banks of both species were nearly depleted, having declined to <5% of their original size. Both species had strikingly different germination strategies: G. lutea seeds mainly germinated in their first-year, whilst the majority of V. album seeds germinated in their second-year. The fraction of dormant G. lutea seeds increased with seed age, indicating that seeds remained viable after forgoing germination in the previous year. Habitat-specific differences in seed germination increased with seed age, with germination fractions being lowest in moist hay meadows. This suggests that the negative effects of anoxic conditions became more pronounced as seeds aged in hay meadows. Conversely, seed dormancy was equal among habitats. The absence of a long-term persistent seed bank has important implications for the management of both nuisance and endangered-plant populations. In the case of V. album and G. lutea, re-colonization of habitats from the seed bank is unlikely after established plants have been removed.

Unpalatable plants facilitate tree sapling survival in wooded pastures

Summary In endangered wooded pasture ecosystems established tree saplings are frequently found in spatial association with protective structures, suggesting nurse effects. This associational resistance is thought to be a driving force behind tree regeneration in wooded pastures. Experimental evidence for associational resistance is, however, scarce. We studied the effects of unpalatable plants on tree sapling performance in a wooded pasture ecosystem in Switzerland and tested the associational resistance hypothesis experimentally. We transplanted 600 Picea abies saplings of two size classes (5 cm and 10 cm) in plots: (i) with unpalatable plants (either Gentiana lutea or Cirsium acaule, representing a chemically and mechanically defended species, respectively); (ii) with one of both unpalatable plants clipped to the ground; and (iii) without any unpalatable plants. Grazing intensity was measured once in each plot and tree sapling performance was followed for 2 consecutive years. Grazing intensity was significantly higher in plots without unpalatable plants, and significantly higher in plots with Cirsium than with Gentiana. Correspondingly, sapling survival (21·5% and 10·6% after 1 and 2 grazing years, respectively) was significantly higher near unpalatable plants, and significantly higher in plots with Gentiana than with Cirsium. Large tree saplings survived significantly better than small ones, and depended less on the unpalatable plants for survival. Except for saplings planted near Gentiana, sapling removal by cows and horses was the major cause of death, the extent varying between unpalatable species and treatments. The growth in height of the saplings was unaffected by the treatments. Synthesis and applications. This study shows that unpalatable plants can enhance tree regeneration in wooded pastures. Sapling survival was significantly higher near unpalatable plants where grazing intensity was significantly lower. Protection against cattle is particularly important for small saplings. These results have important management implications for the endangered and disappearing wooded pastures in western Europe. Transplanting tree saplings near unpalatable plants could be an alternative reforestation technique in intensively grazed wooded pastures. On the other hand, removal of unpalatable plants and shrubs might prevent or slow down undesired tree encroachment in less intensively grazed areas.

Effects of Gentiopicroside, Sweroside and Swertiamarine, Secoiridoids from Gentian (Gentiana luteassp. symphyandra), on Cultured Chicken Embryonic Fibroblasts

Wound healing properties of Gentian (Gentiana lutea ssp. symphyandra) extract and its main constituents, gentiopicroside, sweroside and swertiamarine (compounds 1-3, respectively) were evaluated by comparison with dexpanthenol on cultured chicken embryonic fibroblasts. The extract was also analyzed by HPLC to quantify its constituents. Chicken embryonic fibroblasts from fertilized eggs were incubated with the plant extract and its constituents, compounds 1-3. Using microscopy, mitotic ability, morphological changes and collagen production in the cultured fibroblasts were evaluated as parameters. Wound healing activity of Gentian seems to be mainly due to the increase in the stimulation of collagen production and the mitotic activity by compounds 2 and 3, respectively (p < 0.005 in all cases). All three compounds also exhibited cytoprotective effects, which may cause a synergism in terms of wound healing activity of Gentian. The findings demonstrated the wound healing activity of Gentian, which has previously been based only on ethnomedical data.