Yield Of Total Alkaloids Research Articles

Plant growth and phytoactive alkaloid synthesis in kratom [Mitragyna speciosa (Korth.)] in response to varying radiance.

Leaves harvested from kratom [Mitragyna speciosa (Korth.)] have a history of use as a traditional ethnobotanical medicine to combat fatigue and improve work productivity in Southeast Asia. In recent years, increased interest in the application and use of kratom has emerged globally, including North America, for its potential application as an alternative source of medicine for pain management and opioid withdrawal syndrome mitigation. Although the chemistry and pharmacology of major kratom alkaloids, mitragynine and 7-hydroxymitragynine, are well documented, foundational information on the impact of plant production environment on growth and kratom alkaloids synthesis is unavailable. To directly address this need, kratom plant growth, leaf chlorophyll content, and alkaloid concentration were evaluated under three lighting conditions: field full sun (FLD-Sun), greenhouse unshaded (GH-Unshaded), and greenhouse shaded (GH-Shaded). Nine kratom alkaloids were quantified using an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Greenhouse cultivation generally promoted kratom height and width extension by 93–114% and 53–57%, respectively, compared to FLD-Sun. Similarly, total leaf area and leaf number were increased by 118–160% and 54–80% under such conditions. Average leaf size of plants grown under GH-Shaded was 41 and 69% greater than GH-Unshaded and FLD-Sun, respectively; however, no differences were observed between GH-Unshaded and FLD-Sun treatments. At the termination of the study, total leaf chlorophyll a+b content of FLD-Sun was 17–23% less than those grown in the greenhouse. Total leaf dry mass was maximized when cultivated in the greenhouse and was 89–91% greater than in the field. Leaf content of four alkaloids to include speciociliatine, mitraphylline, corynantheidine, and isocorynantheidine were not significantly impacted by lighting conditions, whereas 7-hydroxymitragynine was below the lower limit of quantification across all treatments. However, mitragynine, paynantheine, and corynoxine concentration per leaf dry mass were increased by 40%, 35%, and 111%, respectively, when cultivated under GH-Shaded compared to FLD-Sun. Additionally, total alkaloid yield per plant was maximized and nearly tripled for several alkaloids when plants were cultivated under such conditions. Furthermore, rapid, non-destructive chlorophyll evaluation correlated well (r2 = 0.68) with extracted chlorophyll concentrations. Given these findings, production efforts where low-light conditions can be implemented are likely to maximize plant biomass and total leaf alkaloid production.

Optimization of Extraction Process for Total Alkaloids from Maca with Different Color Types

This study was carried out to optimize the acidic extraction of total alkaloids from Maca with different color type. The effects of three extraction parameters including solid-to-liquid ratio, temperature and duration on extraction efficiency were investigated by orthogonal array design. Results were as follows:The order of effect of different factors on extraction efficiency for yellow Maca and black Maca were solid-to-liquid ratio > duration > temperature. The order of effect of different factors on extraction efficiency for purple Maca were duration > solid-to-liquid ratio > temperature. The content of total alkaliods in yellow Maca was 9.89%±0.38% under the optimum extracting conditions:Ratio of solid to liquid 1:30, temperature 75℃, time 5.5 h.The optimal extraction condition of purple Maca were 90℃ temperature for 5.5 h with solid-to-liquid ratio 1:25, and the yield of total alkaloids was 10.36%±0.58%. The optimal extraction parameters for black Maca was as follows:Temperature 85℃, time 6 h, solid-liquid ratio 1:20, under these conditions, the extraction yield of total alkaloids was 10.01%±0.78%. In conclusion, the optimal extraction conditions for total alkaloids in different color types Maca had some differences.

Qualitative phytochemical screening and antimicrobial evaluation of the total alkaloids of the hydroalcoholic extracts of the leaves and stem bark of Musanga cercropoides (Urticaceae)

Background: Musanga cecropioides R.Br. ex Tedlie (Urticaceae) is a plant that is widely used in Nigerian traditional medicine for the treatment of various infections such as cough, diarrhoea, and vaginal candidiasis. Objective: To study the antimicrobial potency of the total alkaloids of the hydroalcoholic extracts (HAE) of the stem bark (ST) and leaves of Musanga cecropioides. Method: The antimicrobial activities of the total alkaloidal constituent of the HAE of the ST and leaves were screened using agar diffusion method using Staphylococcus aureus, Bacillus subtils,Klebsiella pneumonia, Escherichia coli, and Candida albicans as test organisms. The preliminary phytochemical screening of the ST and leaves were performed following the conventional standard procedures of Trease and Evans. Results: Phytochemical screening revealed the presence of alkaloids, phenols, flavonoids, tannins, sterols, saponins and terpenoids. The HAE of both ST and leaves exhibited notable activities against the tested microorganisms at different concentrations (500, 250, 125, 62.5 mg/ml) with zones of inhibition ranging from 8.6±0.5mm to 20.0±1.0mm. The highest activity against bacteria had a zone of inhibition of 18.6±1.1mm and 20.0±1.0mm for fungus. The zone of inhibition for antibacterial control drug (Ciprofloxacin) was 42.3±2.0mm while that of antifungal control drug (Griseofulvin) was 32.0±0.8mm. The total alkaloid yield for the ST was 1.200%w/w and 1.332%w/w for the leaves. Conclusion: Musanga cercropoidesmay be a valuable source of a therapeutic agent for potent clinical antimicrobials.

Effect of Nitrogen, Phosphorus and Medium pH to Enhance Alkaloid Production from Catharanthus roseus Cell Suspension Culture

Several elevated levels of nitrogen and phosphate at varying pH of the medium which impart a major influence on callus and biomass development and subsequent production of alkaloids was investigated using suspension culture system of Catharanthus roseus in the present study. The B5 medium was buffered at pH 4.51, 5.82 and 7.32 by addition of different levels of (A) diammonium hydrogen phosphate (NH4)2HPO4 and (B) ammonium dihydrogen orthophosphate (NH4H2PO4) representing the enhanced and varied supply of total nitrogen (NH4++NO3ˉ) and phosphate compared to MS medium (as control) for cell biomass production and alkaloid yield. The pH of the medium have shown significant effects with maximum biomass fresh wt., dry wt. and total alkaloid yield at 5.82 medium pH with elevated phosphate levels and total nitrogen concentration of 3710.10 mg/L compared to control MS medium with 2850 mg/L total nitrogen. At 3667.33 and 3752.48 mg/L of total nitrogen with enhanced phosphate supply showed reduced biomass fresh wt., dry wt. and total alkaloid yield at lower (4.51) and higher (7.32) medium pH respectively. Inclusion of 200 mg/L of tryptophan or phenylalanine as reduced nitrogen source in B5 medium buffered at 5.82 ± 0.2 pH showed enhanced biomass and alkaloid production. Hence, addition of nitrogen, phosphate, tryptophan, phenylalanine as nutrient in suspension culture stimulate their uptake to enhance cell biomass and total alkaloids production but as a function of pH of the medium.

High Performance Liquid Chromatography Determination and Optimization of the Extraction Process for the Total Alkaloids from Traditional Herb Stephania cepharantha Hayata.

Stephania cepharantha Hayata is a traditional Chinese herbal medicine used to treat lung cancer, and its alkaloids, especially cepharanthine (CEP), were reported to be its effective ingredients. Therefore, the extraction of potential antitumor ingredients from the plant was of interest. We first explored the optimized solvent extraction of antitumor agents from S. cepharantha Hayata guided by an in vitro antitumor activity assay. The solvent for extraction and its concentration, the liquid to material ratio, extraction duration, particle size, macerating time, and the frequency of extraction were investigated using a single-factor experiment. An orthogonal design (L9, 34) was constructed to determine the suitable extraction conditions. The crude extract was then purified sequentially by macroporous adsorption resins (MR) for the enrichment of CEP. Under these optimal conditions, the yield of total alkaloids in the herbs was 3.4%, whereas the CEP content was 2.9%. Total alkaloids exhibited significant anti-proliferative activities in the A549 cell line. Our study provides means for the further development and use of the antitumor components from S. cepharantha, which has potential for application in the pharmaceutical industry.

Heterosis and heterobeltiosis studies on yield and yield components of some Turkish poppy hybrids (Papaver somniferum L.)

Poppy (Papaver somniferum L.) is highly strategic and sensitive economic crop. A set of 7 potential poppy parent lines and cultivars along with their 42 reciprocal hybrids was made to collect information on extent of heterosis and heterobeltiosis for selection of better parents for capsules, seeds, morphine and total alkaloids yield. The study was carried out during 2012–13 and 2013–14 growing seasons. Heterosis and heterobeltiosis values of capsule yield varied between −19.59% to 94.87% and −28.84% to 87.71%, seed yield varied between −14.75% to 83.61% and −21.21% to 78.67%, morphine yield varied between −26.87% to 174.35% and −32.68% to 137.70% total alkaloid yield ranged −23.92% to 160.50% and −32.46% to 145.67% respectively. It was noted that TMOT×Ofis-8 and TMOTד2010” were the best hybrids in terms of capsule, seed, morphine and total alkaloid yield. Whereas, the maximum heterotic depression for capsules, seeds, morphine and alkaloid yield was noted for N-442×TMOT hybrids. The study showed good scope for commercial exploitation of heterosis and isolation of pure lines among progenies of heterotic F1 hybrids. Thus, taking an overview of the results of this study it can be suggested that, future studies should focus on TMOT, Ofis-8, “2010” and N 442 cultivars for breeding new high and low morphine cultivars in hybridization programs profitably.

Sink ? Source System of In vitro Suspension Culture of Celastrus paniculatus under Regulation of Monochromatic Lights

Plants are wonderful resource of bioproducts encompassing significant value to medicines and drug development. The plant cell suspension cultures bear immense potential for production of high?value secondary metabolites and are chosen as alternative sources of raw material for industrial use. In the present study, homogenous cell suspension culture of Celastrus paniculatus a medicinally important plant was established and multifold production of alkaloids and total phenols was obtained under the influence of monochromatic lights. One month old leaf derived friable callus of C. paniculatus was used to raise homogenous suspension culture and kept on rotary shakers in cabinets illuminated with different monochromatic LED lights (Blue, Yellow and Red). The monochromatic lights proved to be a strong abiotic elicitor in driving the production of secondary metabolites so much so that the metabolites were released extracellularly and the medium served as sink or spacious pool for leaked out metabolites from the cell mass. Maximum production and enhancement in alkaloids and phenols (98 and 44.7%, respectively) over control was obtained from cell mass grown under yellow light treatment, followed by blue (64 and 23.7%) and red light (50 and 26%) treatments. Further scale up of secondary metabolite production was hence performed under yellow light conditions, starting from 2.5 gm cell mass suspended in 250 ml of media extended up to 1000 ml culture media for one month. The continuous culture system exhibited remarkable potential of this plant cell system as multifold yield of total alkaloids (91.69 ?g/1750 ml) and phenols (70.59 ?g/1750 ml) was obtained during 30 days of culture under yellow light conditions. The production was remarkably enhanced by 100?folds (5.29 to 48.21 ?g; Fig. 2) for alkaloids and 70?folds (1.73 to 46.33 ?g; Fig. 3) for phenols, from zero days to 30?day?old culture phase. Hence, strategic implementation of monochromatic lights holds great promises for controlled commercial production of myriads of valuable secondary metabolites from plant cell cultures of important medicinal plants. Present work contributes to the first reports where continuous, enhanced, multifold yield of important secondary metabolites were obtained from cell suspension culture of Celasrus paniculatus an endangered medicinal plant.Plant Tissue Cult. & Biotech. 26(2): 175-185, 2016 (December)

Effects of Sowing Date on Some Agronomic Characteristics and Alkaloid Content of Datura Stramonium in Semi-Arid Conditions

Abstract Alkaloids of diferent plant, including many Solanaceae species, constitute important natural sources for variety of pharmaceutical products. The amounts of various secondary plant products are strongly dependent on the growing conditions and agronomical applications such as planting date, fertilization, irrigation etc. The aim of this was to study the effects of sowing date on some agronomic characteristics and total alkaloid content of thorn apple (Datura stramonium), investigated in the environmental conditions from Dicle University, Agricultural Faculty, Field Crops Department experimental area, during 2010 and 2011 growing years. In the research, plant height, stem diameter, number of branches per plant, number of capsule per plant, capsule width, capsule length, 1000-seed weight, fresh herb yield, herba yield, seed yield and total alkaloid content were examined. At the end of the study, in the trial of thorn apple with sowing dates, according to two years mean, seed yield, fresh herb yield, dry herb yield and total alkaloid yield were changed between 335 kg ha−1 and 704 kg ha−1, 5933 and 20537 kg ha−1, 1613 kg ha−1 and 4800 kg ha−1, and 0.270% and 0.391%, respectively. The effect of sowing date was found significant on the investigated characteristics, when sowing delayed, agronomic characteristics were also decreased. The highest values related with seed yield, fresh and dry herbage of thorn apple were obtained sowing of 01 Apr. Moreover, thorn apple plant showed morphogenetic variation when compared leaf and seed alkaloid contents.

English

The vegetative parameters of ashwagandha (viz. plant height, number of primary branches, plant spread and dry weight of shoot) were enhanced significantly with the application of 12.5 mg P2O5 kg-1 soil, whereas; dry weight of roots was enhanced up to the application level of 25 mg P2O5 kg-1 soil. FYM at the rate of 12.5 t ha-1 in combination with 12.5 mg P2O5 kg-1 soil significantly improved all the vegetative parameters, whereas, FYM at the rate of 12.5 t ha-1 in combination with 25 mg P2O5 kg-1 soil significantly enhanced the dry weight of the roots. The alkaloids yield (mg pot-1) in ashwagandha roots increased significantly with the application of 25 mg P2O5 kg-1 soil. The application of FYM at the rate of 12.5 t ha-1 improved the alkaloids yield (mg pot-1) but the significantly highest yield of total alkaloids (mg pot-1) was found in the treatment combination of 12.5 t FYM ha-1 + inorganic-P at the rate of 25 mg P2O5 kg-1 soil as compared with other eleven treatments. Nutrients (NPK) uptake by ashwagandha shoot increased significantly with the application level of 12.5 mg P2O5 kg-1 soil over control whereas in case of ashwagandha roots, the increase in nutrients uptake at the level of 25 mg P2O5 kg-1 soil over control. Application of FYM at the rate of 12.5 t ha-1 + 12.5 mg P2O5 kg-1 soil was the best treatment combination for ashwagandha roots in terms of nutrients uptake. Application of fertilizer-P significantly improved the status of organic carbon, Available-P in post harvest soil but decreased the Available-N, Available-K and DTPA extractable micronutrients (Fe, Cu, Zn and Mn) up to the level of 25 mg P2O5 kg-1 soil.   FYM at the rate of 12.5 t ha-1 helped in maintaining the soil fertility status after harvest of the crop alone or in combination with fertilizer-P.   Key words: Ashwagandha, phosphorus, farmyard manure, alkaloids, nutrients uptake, soil fertility.

Ashwagandha (Withania somniferaL. Dunal) crop as Affected by the application of FYM and inorganic phosphorus in typic torripsamments of Hisar

The vegetative parameters of ashwagandha (viz. plant height, number of primary branches, plant spread and dry weight of shoot) were enhanced significantly with the application of 12.5 mg P2O5 kg−1 soil, whereas; dry weight of roots was enhanced up to the application level of 25 mg P2O5 kg−1 soil. FYM at the rate of 12.5 t ha−1 in combination with 12.5 mg P2O5 kg−1 soil significantly improved all the vegetative parameters, whereas, FYM at the rate of 12.5 t ha−1 in combination with 25 mg P2O5 kg−1 soil significantly enhanced the dry weight of the roots. The alkaloids yield (mg pot−1) in ashwagandha roots increased significantly with the application of 25 mg P2O5 kg−1 soil. The application of FYM at the rate of 12.5 t ha−1 improved the alkaloids yield (mg pot−1) but the significantly highest yield of total alkaloids (mg pot−1) was found in treatment combination of 12.5 t FYM ha−1 + inorganic-P at the rate of 25 mg P2O5 kg−1 soil as compared with other eleven treatments. Nutrients (NPK) uptake by ashwagandha shoot increased significantly with the application level of 12.5 mg P2O5 kg−1 soil over control whereas in case of ashwagandha roots, the increase in nutrients uptake at the level of 25 mg P2O5 kg−1 soil over control. Application of FYM at the rate of 12.5 t ha−1 + 12.5 mg P2O5 kg−1 soil was the best treatment combination for ashwagandha roots in terms of nutrients uptake. Application of fertilizer-P significantly improved the status of organic carbon, Available-P in post harvest soil but decreased the Available-N, Available-K and DTPA extractable micronutrients (Fe, Cu, Zn and Mn) up to the level of 25 mg P2O5 kg−1 soil. FYM at the rate of 12.5 t ha−1 helped in maintaining the soil fertility status after harvest of the crop alone or in combination with fertilizer-P.

Microbe associated molecular patterns from rhizosphere bacteria trigger germination and Papaver somniferum metabolism under greenhouse conditions

Ten PGPR from different backgrounds were assayed on Papaver somniferum var. Madrigal to evaluate their potential as biotic elicitors to increase alkaloid content under the rationale that some microbe associated molecular patterns (MAMPs) are able to trigger plant metabolism. First, the 10 strains and their culture media at two different concentrations were tested for their ability to trigger seed germination. Then, the best three strains were tested for their ability to increase seedling growth and alkaloid levels under greenhouse conditions. Only three strains and their culture media enhanced germination. Then, germination enhancing capacity of these best three strains, N5.18 Stenotrophomonas maltophilia, Aur9 Chryseobacterium balustinum and N21.4 Pseudomonas fluorescens was evaluated in soil. Finally, the three strains were applied on seedlings at two time points, by soil drench or by foliar spray. Photosynthesis was measured, plant height was recorded, capsules were weighted and alkaloids analyzed by HPLC. Only N5.18 delivered by foliar spray significantly increased plant height coupled to an increase in total alkaloids and a significant increase in opium poppy straw dry weight; these increases were supported by a better photosynthetic efficiency. The relative contents of morphine, thebaine, codeine and oripavine were affected by this treatment causing a significant increase in morphine coupled to a decrease in thebaine, demonstrating the effectivity of MAMPs from N5.18 in this plant species. Considering the increase in capsule biomass and alkaloids together with the acceleration of germination, strain N5.18 appears as a good candidate to elicit plant metabolism and consequently, to increase productivity of Papaver somniferum.

Influence of Soil Sodicity on the Growth, Alkaloid Yield, and Cation Accumulation of Catharanthus roseus

The effects of soil sodicity (exchangeable sodium percentage, or ESP) on the growth, alkaloid yield, and cation accumulation of two cultivars of Catharanthus roseus (Nirmal and Dhawal) was studied in a pot experiment. The leaves and stem yield of C. roseus significantly increased with an increase in soil ESP from 4.3 (control) to 16.9, but thereafter yield decreased with further increases in soil ESP. The severe leaf injury symptom from soil sodicity was exhibited in both cultivars at 32 weeks after transplanting into soil at ESP levels of 44.9 and above. Root yields of C. roseus were also significantly reduced with an increase in soil ESP. Total alkaloid yield in leaves increased with increases in soil at ESP levels of 32.0 and 16.9 for the cultivars Nirmal and Dhawal, respectively, but further increases in soil ESP the total alkaloid yield significantly decreased. The vindoline content in leaves of C. roseus significantly decreased with an increase in soil ESP. In the shoot and root tissues of C. roseus, the concentration of sodium significantly increased and that of potassium, and calcium decreased in plants grown in soil with high sodicity (ESP 44.9 and above) as compared with the control.

Alkaloids of Buxus colchica, B. sempervirens, and B. balearica growing in Georgia

The introduced species Buxus sempervirens and B. balearica grow in Georgia in addition to the endemic B. colchica Pojark. We studied leaves and stems of these plants collected during flowering and fruiting near Tbilisi, in Batum Botanical Garden, and on Shaor Pass. Pharmacological tests of total preparations that were performed at the Kutateladze Institute of Pharmaceutical Chemistry showed that total alkaloids soluble in ether, CHCl3, and water exhibited spasmolytic activity. The strength of the activity of the aqueous extracts of the Buxus species could be placed in the following order [1]: B. balearica Lam > B. sempervirens > B. colchica Pojark. Plant material was extracted beforehand with EtOAc followed by extraction of alkaloids by diethylether and CHCl3 after making the raw material basic in order to obtain total alkaloids. Total alkaloids soluble in water were obtained as an extract from leaves and stems of B. colchica, B. sempervirens, and B. balearica according to requirements of the SP, XIth Ed. (1982, Vol. II, p. 160). Seasonal variations of alkaloids in vegetative organs were studied by obtaining total alkaloids in a Soxhlet apparatus using extraction by CHCl3 and basic raw material with subsequent acid–base purification and separation into ether and CHCl3 fractions. Total alkaloids soluble in ether were separated using citrate–phosphate buffers in pH steps 6.6, 6.0, 5.0, 4.0, and 3.0 with 0.1 M citric acid. Separation into pure bases was carried out over Al2O3 (activity II, III, VI, and neutral) and SiO2 (100/160 μm) columns. The ratio of compounds to sorbent was 1:100. Elution used ether, benzene:ether (1%), benzene:EtOH (1–5%), benzene:CHCl3, CHCl3:CH3OH with increasing MeOH content. Fractions of 25–50 mL were collected. The yield of total alkaloids was determined by acid–base titration calculated as molecular weight of buxamine; the buxamine content, by a chromatographic titration method using B. colchica and B. sempervirens. Qualitative analysis was performed in thin layers of SiO2 (Merck) on plates using MeOH:hexane:NH4OH (25%) (40:5:3). The yield of total alkaloids from B. balearica was determined gravimetrically [2]. Alkaloids were identified based on a determination and analysis of physicochemical (mp, [α]D) and spectral data (UV, IR, PMR, mass) in addition to comparison with the literature [1-9]. Reference samples for TLC mobility were authentic alkaloids. Table 1 shows the variations in alkaloid contents with development phases in the vegetative organs. B. balearica, yield of total alkaloids from leaves and stems during flowering, 6%. The fraction from pH 0.6; 6.5 of the ether total was separated over an Al2O3 column with elution by benzene:CHCl3 and benzene:EtOH (1→5%). Fractions from EtOH (72–126) afforded 1, mp 199–201°C (EtOH), [α]D –100.3° (CHCl3). Mass spectrum (m/z): 414 [M]+, 386, 371, 356, 343, 340, 84, 72 (100), 58. PMR spectrum (CDCl3, δ, ppm): 0.74 (3H, s, CH3), 0.88 (6H, s, 2 × CH3), 0.92 (6H, s, 2 × CH3), 2.2 [6H, s, N(CH3)2], 2.40 [6H, s, N(CH3)2]. The experimental results allowed us to identify the isolated compound as cycloprotobuxin-A [4, 5]. B. sempervirens, yield of total alkaloids, 2.2%. Amorphous base 2 was isolated from the fraction with pH 5.0 of the ether total alkaloids by separation over a column of Al2O3 with elution by benzene:ether (1%) from effluents (9–12).

Enhanced Extraction of Alkaloids from Sophora alopecuroides L. by Ion Exchange at Reduced Pressure

The extraction of total alkaloids from Sophora alopecuroides L. was enhanced by using ion exchange resin at reduced pressure. At room temperature and 0.08 MPa vacuity for 12 h, 96% of the total alkaloids was extracted from the seed particles of 0.18–0.25 mm using deionized water of pH 3.0 in the presence of the cationic ion exchange resin 001×7 (cross-linked polystyrene, particle size 0.32–1.25 mm, exchange capacity 4.5 mmol g–1 resin, dry wt.). In contrast, it took the same time to get 72% of total alkaloids in the case without the resin. The yield of total alkaloids extracted at 0.06 MPa vacuity without the resin reached 60%, while that was only 47% in the case at normal pressure.

Chemical Composition and Pharmacological Activity of Alkaloids from the Common Periwinkle Cultured in Georgia

The common periwinkle (Vinca minor L.) is a perennial herbaceous plant used for a long time in both classical and folk medicine for the treatment of various disorders [1, 2]. The main set of biologically active substances in this plant is represented by indole alkaloids, predominantly of the eburnamine group. It was experimentally established that alkaloids of this group favorably affect the cerebral circulation, producing a more than twofold increase in the blood flow rate [3]. Experience gained with foreign preparations based on this plant such as oxibral and vincanor (vincapan, vincaton) confirmed that the common periwinkle is a valuable medicinal plant [1, 4]. In Georgia, the common periwinkle was cultivated as a medicinal plant on an experimental field of the Institute of Pharmaceutical Chemistry at the Vashla-dzhvari village near Tbilisi. The green mass yield from the experimental field was 2 – 2.5 kg m. An attempt at transferring the common periwinkle to the field of the Experimental Center of Medicinal Plants (Shiraki, East Kakhetia) was successful, leading to an increase in the alkaloid content. The total alkaloid yield reached up to 1 and 0.8%, in which the vincamine content amounted to 0.06 and 0.04%, in the blooming and fruitage stages, respectively. The purpose of this study was to refine the chemical composition of alkaloids from the common periwinkle cultivated in Georgia and check for the pharmacological identity of the vincamine base and the active alkaloid fraction extracted from herbs. The experiments were performed with an air-dry material prepared from plants collected in the blooming period (May – April) in 1997 and 1998 on the experimental field at the Institute of Pharmaceutical Chemistry (Tbilisi). The main stages of the total alkaloid extraction and the isolation of individual alkaloid fractions are indicated in the scheme. The course of the extraction process was monitored and the homogeneity of products checked by TLC. All TLC runs demonstrated spot patterns with reproducible Rf values. Pharmaceutical Chemistry Journal Vol. 35, No. 5, 2001

Pharmacognosy of Catharanthus roseus (L.) G. Don (Sadabahar) Root and its Adulterants

AbstractCommercial samples of Catharanthus roseus G. Don roots (Sadabahar) are often found adulterated with certain Solanaceous roots like Solanum melongena and Lycopersicon esculentum which adversely affect the quality as well as total alkaloid yield of the drug. In order to detect the contaminants in authentic drug samples, a detailed pharmacognostic study was carried out which highlighted certain diagnostic macro- and microscopic characters useful for their identification. Macroscopic characters of roots of white and pink forms of C. roseus are distinct from those of their adulterants in form, color and odor. Microscopically, both forms of C. roseus are distinguished mainly on the basis of size of starch grains and vessels. Detailed microscopic features of the authentic drug and the adulterants exhibit certain important features of pharmacognostic interest. The bark of the root of C. roseus is rich in alkaloids. Onadry weight basis, yield of wood per 100 g of root is about 1.5 times more than the bark ...

Ergot Alkaloids and Some Problems of the Physiology of Their Formation

Alkaloids cannot be supposed to be waste products of general metabolism. They are formed only for a relatively brief time during the lifespan of the culture and under conditions of reduced proliferation. Living rather than dying cells are required for alkaloid synthesis. The alkaloid yield and differentiated cells were observed to be inversely proportional. This pattern is correlated with formation of secondary shunt products. The district between the cell-pool tryptophan “down” and “up” phases is a promising target for the investigation of regulation of the alkaloid formation and continuous fermentation of these compounds. Considering the general phenomenon of secondary metabolism we may guess that alkaloid formation includes derepression of the appropriate portion of the genome to obtain transcription and translation of specific synthetases and subsequent activity of these enzymes for a relatively brief period. The ability to produce alkaloids is not only easily lost genetically, but is also extraordinarily susceptible to suppression by rather minor environmental manipulations. The production of alkaloids occurs only in nutrient media with high osmotic pressure. Ergoline alkaloids do not definitely belong to a group of physiologically inert secondary metabolites. They are, in fact, able to influence primary shunt metabolism, and may reflect a regulatory device of the producing cell. Alkaloids increase the activities of tryptophan synthetase and acetyl-CoA carboxylase of the ergot fungus and exert a negative effect on citrate synthase, isocitrate lynse, and malate synthase. Also inhibition of Claviceps peptidases by peptide alkaloids is of peculiar interest. Addition of ergometrine (10 -4 M) to the submerged culture of C. paspali increased the total-alkaloid yield by 120%. However, the stimulatory effect was inversely proportional to the concentration of the supplied ergometrine. Implicit in these data is that in alkaloid-producing cultures accumulating alkaloids may suppress the attainment of the yield that would be achieved by continuous control of the alkaloid level. Man weiß seit langem, daß im allgemeinen Mutterkornzuchtstämme, die unter parasitischen Bedingungen reichlich Alkaloide bilden, bei der saprophytischen Kultivation keine oder nur in Spuren Ergoline synthetisieren. Die vorliegende Übersicht ist ein Beitrag zur Problematik der Physiologie der Alkaloidbildung bei saprophytischen Claviceps -Arten. Die Mutterkornalkaloide treten nur in bestimmten Entwicklungsstadien der Kultur und unter Bedingungen der reduzierten Proliferation auf. Meistens ist die Alkaloidsynthese mit herabgesetzter Zelldifferenzierung verknüpft. Die Fähigkeit des Stammes, Alkaloide zu bilden, ist stark abhängig vom eingesetzten Medium. Die Schlüsselsubstanz aller Ergoline ist das Tryptophan. Der Abschnitt zwischen der Phase des Absinkens und der Phase des Anstiegs des freien Zelltryptophans ist anscheinend ein vielversprechender Ansatzpunkt für die Steuerung der Alkaloidsynthese und für die kontinuierliche Fermentation der Alkaloide. Die Mutterkornalkaloide beeinflussen einige wichtige Enzyme des primären Stoffwechscls, z. B. stimulieren sie die Tryptophansynthetase und die Acetyl-CoA-Karboxylase und hemmen die Citratsynthetase, Isocitratlyase, Malatsynthetase und die intracellulären Exopeptidasen der Claviceps -Stämme. Nach dem Einsatz von Ergometrin (10 -4 M) in die Claviceps -Tiefkultur steigt die Alkaloidausbeute um 120%. Der Effekt hängt von der Menge des zugegebenen Ergometrins ab. Man kann annehmen, daß die Mutterkornalkaloide physiologisch aktive Metabolite darstellen und sich an der Regulation ihrer eigenen Synthese beteiligen.

Physiological activities of ergoline alkaloids in submerged cultures of Claviceps paspali and Claviceps purpurea.

Ergoline alkaloids do not belong to a group of physio ogically inert secondary metabolites. Ergometrine as well as ergotamine increased the acetyl-CoA carboxylase activity and inhibited the activities of citrate synthase and isocitrate lyase. The slightly inhibitory effect of ergotamine and stimulatory effect of ergometrine on malate synthase was established. Supplied ergometrine brought about a change in the culture growth rate,i.e. inhibition (10−4 m) or stimulation (10−3 m). Addition of 10−4 m ergometrine to the culture ofClaviceps paspali MG-6 increased the total alkaloid yield by 120%. The stimulatory influence was inversely proportional to the concentration of added alkaloid. It is assumed that accumulating alkaloids suppress attainment of the yield that would be achieved by continuous control of the alkaloid level. A positive relationship is assumed between the process of alkaloid synthesis and the viability of the culture.

Zur Physiologie der Alkaloidbildung bei Claviceps-Arten

Summary Our strain pepty 695 of Claviceps purpurea produces clavines, ergometrine and ergotoxine alkaloids under submerged conditions. Clearly a “balanced phase”, “transition phase“ and “maintenance phase” of growth and metabolism according to Borrow et al. can be distinguished. Claviceps paspali strain Li 342 follows the fermentation pattern No.2 according to Taber. There was practically no increase in the total alkaloid yield of the ergotoxine strain after the addition of tryptophan or analogues to the preculture or fermentation medium. D, L-4- methyl-tryptophan caused an inhibition of the alkaloid formation. The metabolism of tryptophan under different conditions in C. paspali was investigated. Protein-bound tryptophan is used for alkaloid synthesis. There is a remarkable difference in the utilization of tryptophan in the pre culture and fermentation mycelium. Most of the administered tryptophan is converted during the alkaloid formation phase equally to alkaloids and to 2,3-dihydroxybenzoic acid. The metabolism of valine, leucine and proline in the ergotoxine strain was investigated during different phases of development. A considerable protein turnover during the alkaloid production phase was observed like in other strains. Inhibitors of protein synthesis did not affect the formation of the peptide portion of ergotoxine alkaloids. These results seems to indicate that the synthesis of the peptide portion is an non-ribosomal process. Also the “ergoline synthetase“ is not inhibited by adding cycloheximide to C. paspali cultures.

The alkaloids of the genus Datura, section Brugmansia. I. D. cornigera Hook.

Abstract Hyoscine has been isolated as the main alkaloid from the leaves, flowers, pericarp and seeds of Datura cornigera Hook. Noratropine occurs in the leaves, flowers and pericarp. The following alkaloids have been obtained from the roots, (—)-3α,6β-ditigloyloxytropane, 7-hydroxy-3,6-ditigloyloxytropane, hyoscine, hyoscyamine, atropine, norhyoscyamine, noratropine, 3α,6β-dihydroxytropane and the presence of others indicated. Hyoscyamine is the principal alkaloid of the whole roots but the root-wood, although giving a low yield of total alkaloids, contains a relatively high proportion of noratropine.