Extraction Of Terpenoids Research Articles

Establishing Gerger (<i>Eruca sativa</i>) Leaves as Functional Food by GC-MS and <i>In-vitro</i> Anti-lipid Peroxidation Assays

Conventional food that safeguards against chronic illnesses is known as a functional food. Establishing functional foods starts with phytoconstituent analysis and in-vitro characterization of health benefits. Eruca sativa, popularly known as gerger or jarjeer in Saudi Arabia, is an annual edible shrub cultivated worldwide. Gerger leaves are consumed raw in salads and have additional health benefits. This study investigated the phytochemical profile of aqueous decoction of gerger leaves of the Saudi origin by GC-MS assay. We also performed in-vitro anti-lipid peroxidation and total antioxidant capacity assays using gerger decoction. Twenty-seven chemical compounds belonging to seven classes constituted the gerger decoction: organic siloxanes (39.75%), organic silyl esters (18.28%), phenolics (17.87%), aromatic and aliphatic esters (10.48%), terpenoids (7.09%), heterocycles (3.83%), and sulfur compounds (2.70%). This study reported the presence of compounds mentioned above for the first time in gerger leaves. The decoction method was efficient in the extraction of heat-stable terpenoids like astaxanthin (2.23%), cilonasterol (1.48%), ingol-12-acetate (0.4%), and phytol (2.98%). The in-vitro anti-lipid peroxidation study demonstrated the ability of gerger decoction to inhibit hepatic lipid peroxidation in a significantly dose-dependent (150 to 400 μg/ml) manner compared to quercetin. A dose of 400 μg/ml of gerger decoction resulted in 68.46 ± 0.01% inhibition of oxidation of hepatic lipids. The total antioxidant capacity of gerger leaves reported as the IC50 of the decoction was 217.90 ± 2.2 μg/ml and also statistically significant. The in-vitro models suggested the antioxidant mechanism of gerger was by hydrogen atom transfer and reduction of metal ions. The study substantiated that gerger is a functional food besides established the phytochemical profile contributing to the antioxidant activity. Given that the gerger decoction has a high silicon content and antioxidants, attempt to determine the bioavailability and to identify the molecular targets are essential to overcome bone disorders and oxidative stress.

Effects of co-administration of Ganoderma terpenoid extract with chloroquine on inflammatory markers and antioxidant status in Plasmodium berghei-infected mice

ObjectiveTo understand the protective effects of Ganoderma terpenoid extract (GTE) against Plasmodium berghei-malarial infection in mice, the present study was carried out to evaluate the effects of GTE in combination with chloroquine disulphate (CQ) on erythrocyte-selected inflammatory markers and antioxidant defense status in P. berghei-infected mice. MethodsP. berghei-infected mice were divided into six groups: infected control (IC) group, administered 1 mL Tween 20; GTE100 and GTE250 groups, administered 100 and 250 mg/kg GTE, respectively; GT100 + CQ and GT250 + CQ groups, co-administered 100 and 250 mg/kg GTE plus 30 mg/kg CQ, respectively; and CQ group, administered 30 mg/kg CQ. A separate group of non-infected mice were given 1 mL Tween 20, and served as a normal control group (NC). Extract and drug were dissolved in Tween 20 and administered orally once daily for 12 consecutive days. At the end of the treatment period, mice were anesthetized with chloroform and sacrificed by cervical dislocation. Plasma was prepared from blood obtained from each mouse. Parameters evaluated at the end of the treatment period include parasitemia, red blood cell count, hematocrit, malondialdehyde (MDA), glutathione (GSH), catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10). ResultsInfected mice treated with a combination of GTE and CQ (GT100 + CQ and GT250 + CQ groups) showed significantly reduced parasitemia levels (P < 0.05) compared to those administered GTE alone as well as IC. Significant improvement in body weight (P < 0.05) was also observed in infected mice treated with a combination of GTE and CQ (GT100 + CQ and GT250 + CQ groups), compared to mice receiving GTE alone (GTE100 and GTE250 groups). Plasma MDA and TNF-α concentrations were significantly lowered, and IL-10 concentration was significantly increased in GT100 + CQ and GT250 + CQ groups, relative to the IC group (P < 0.05). GSH concentration and SOD, CAT and GPx activities were significantly higher in GT100 + CQ and GT250 + CQ groups compared to the GTE100, GTE250, IC and NC groups (P < 0.05). ConclusionData generated in this study showed that GTE enhanced the anti-plasmodial action of CQ in mice through its anti-inflammatory and antioxidant activities.

Screening of in vitro antiviral activity of purified terpenoid extracts of selected sea weeds against chikungunya virus

Currently, the search of novel phytochemicals with unique biological potentialities is a pre-requisite for the designing ideal drugs for the human kind. Sea weeds are bioresources with a broad spectrum of medicinal properties with minimal side effects. Kerala, the Southern state of India reported high incidence of Chikungunya virus (CHIKV) infections in the last several tears. No specific virucidal therapy or effective vaccines are available. This emphasizes the need of searching for phytochemicals as drugs with less cost and more effective. Therefore, an attempt was made in screening purified terpenoid extracts of selected sea weeds as anti-CHIKV potential. In this study the terpenoids composition from the red algae Hypnea musciformis, Kappaphycus alvarezii and Gracillaria dura were identified and analyzed by thin layer chromatography and Gas chromatography- Mass spectrum. The methanolic extract of seaweeds was purified by column chromatography and each fraction was eluted by using petroleum ether and ethyl acetate as solvent combination. The analysis of the purified fraction of H. musciformis and K. alvarezii revealed the presence of 8 terpenoid fractions, and G. dura showed only 4 major components respectively. Vero cell lines were employed in the antiviral assays, infected to CHIKV, and treated with varied doses of purified terpenoid extracts. In the antiviral activity, terpenoid extracts of G. dura showed remarkable and promising EC50 inhibitory effect at 1.25 μg/ml. Further, the terpenoid extracts displayed efficient virucidal activity against CHIKV (inhibit around 90%) with 5 μg/ml dosage. As the last phase, terpenoid extracts added at time intervals of 0, 1, 2, 3 post-infection periods still maintained a significant inhibitory potential against CHIKV viral replication. Thus, the overall study suggests that the terpenoid extracts of G. dura may be effectively used in the prevention and treatment of CHIKV infections. Clinical studies may be warranted for designing a promising new anti-CHIKV drug.

Bioactivity and Apoptotic Efficacy of the Purified Terpenoid Extract from the Moss Brachythecium Buchananii (Hook.) A. Jaeger against MG 63 Cells

Bryophytes are primitive non vascular plants. A little is explored regarding the medicinal effects of bryophytes on carcinoma. This study investigated the biological effects of purified terpenoids from Brachythecium buchananii on selected cell lines such as HeLa, MDAMB-231 and MG63 human osteosarcoma cells and also elucidating the regu­latory signaling pathways underlying the effects of terpenoids towards caspase cascade and the antioxidant enzyme system. The cell lines were treated with various concentrations of purified terpenoid extracts interms of evaluating viability (MTT assay). Interestingly, MG63 cell lines showed poor viability as compared to other ongo cells and was subjected to further molecular evaluations. Migration and invasion assay results using wound-healing and transwell assays, respectively reveal the pro-antimetastatic potential of the purified terpenoids from the moss. The flow cytometry study substantiated terpenoid induced apoptosis in MG63 cells. Cell cycle analysis revealed the significant increase in the number of cells arrested at the S growth phase. Terpenoid extract also displayed DNA fragmentation in the cells. Western blot analysis revealed the down regulation of the anti‑apoptotic proteins Bcl‑2, pro‑caspase 3 and over expression of the pro‑apoptotic protein Bax. In addition, the caspase cascade profile of the terpenoid extract substantiated their efficacy in tumour inhibition. Thus, the overall results confirmed the biological features of terpenoid induced apoptosis in the MG63 cells.

The entangled dynamics of eucalypt leaf and flower volatile emissions

Eucalypts (including Angophora, Corymbia, Eucalyptus) comprise a large and hyper-diverse group of ecologically important trees providing habitats for many forest dependent species. Eucalypts synthesize a plethora of specialized metabolites that moderate biotic and abiotic interactions. Until now, studies on eucalypt volatile organic compounds (VOCs) have primarily focused on isoprene and monoterpenoid emission from forest plantations. As a result, little is known about the volatile profile of adult eucalypt trees under natural conditions.Here we present an in-depth study of leaf and flower branch volatile emissions from mature trees of nine eucalypt species in south-eastern Australia. VOCs were captured using a non-invasive, dynamic flow-through enclosure technique enabling volatile collection into adsorbent cartridges, analyzed by GC–MS. For comparison, terpenoid extracts were also analyzed. Complex emission profiles composed of more than 100 different VOCs were recorded, including isoprene, mono- and sesquiterpenoids, and benzenoids. Emission profiles from eucalypts were highly species-specific, tissue dependent, diurnally regulated and affected by environmental conditions. The composition of stored terpenoids did not mirror the emission profiles, supporting differential regulation of formation, storage and emission of terpenoids. These results provide important knowledge about the entangled dynamics of eucalypt volatiles, and have wider implications for predictions of volatile emission from eucalypt forests and plantations.

Optimization by experimental design of headspace sorptive extraction and solid-phase microextraction for the determination of terpenes in spices

Headspace sorptive extraction (HSSE) was compared with solid-phase microextraction (SPME), a reference method for the extraction of volatiles. HSSE and SPME were optimized by experimental design for the extraction of terpenes and terpenoids from spices. The different extraction parameters studied were the extraction time, the extraction temperature, the sample mass, and the equilibrium time (only for SPME). Extracts were obtained by HSSE and SPME from six spices: cinnamon, cumin, thyme, fennel seeds, nutmeg, and clove. Those extracts were analyzed by gas chromatography coupled to a mass spectrometer (GC-MS) qualitatively and quantitatively. The quantitative analysis was conducted using 28 standards. For each standard, calibration curves, limits of detection (LOD) and quantification (LOQ) were determined. The study shows that HSSE is selective, sensitive, and highly repeatable method for the extraction of volatile terpenes and terpenoids from spices and allows to produce extracts concentrated in the μg/g range of terpenes.

Effect of temperature and different extracts of Datura metel leaves on some biological aspects of Hyalomma schulzei (Acari: Ixodidae)

The current study investigated the effect of temperatures (28, 35 and 40) ° C Under 90 % R.H and alkaloids, phenolic and terpenoid extracts of Datura metel leaves at different concentration on biological performance of Hyalomma schulzei. The result indicated that egg hatchability was 97% at 28 and 35° C., incubation period was 30.66 days at 28° C the premoulting period for larvae and nymphs were (10.7-14.7) and (18.2-23) days respectively. Preoviposition period was 11.33 days at 28 °C while oviposition period was 21days at the same temp. Total egg production was (8848-8850) eggs and female converted 56.1% of engorged weight into eggs irrespective of temp. Bioassay results revealed that crude alkaloids, terpenoids and phenolics compounds of Datura metel leaves have an acaricidal effects on different developmental stages of Hyalomma schulzei. The mortality of eggs and starved larvae was 90% in all used conc. Of alkaloid, terpenoid and phenolics, the same mortality percentage was exhibited by fed larvae in all conc. Of alkaloid and terpenoids, however, mortality rate of starved and fed nymphs were 74.21% and 64.41% respectively in alkaloid extracts only. Moreover, terpenoid and phenolic extracts have no impact on adult females and males mortality. The outcome of the present study proved that alkaloid extracts of Datura metel. Leaves could effectively control different life stages of Hyalomma schulzei

EVALUATION OF THE CRUDE PHENOLIC AND TERPENOID EXTRACTS OF CARISSA MACROCARPA AGAINST APHIS FABAE SCOPOLI (HEMIPTERA: APHIDIDAE) IN-VITRO

ABSTRACT Present study was conducted to evaluate different concentrations of crude phenolic and terpenoid extracts of the leaf Carissa macrocarpa as alternatives to chemical insecticides in some biological aspects of Aphis fabae under in vitro condition. The results showed that all plant extract concentrations were effective against adult of A. fabae compared with control treatment. The mortality rates were of the adult when used 10 mg/ ml concentration of the crude phenolic and terpenoid extracts after 48 hrs. of exposure 46.4 and 52.3% Compared with 9.2 and 13.8% in control treatment respectively. Also, the crude phenolic and terpenoid extracts have affected reduced the number of birth reached 5.33 and 4.67 nymph/ Female at concentration 10 mg/ mL of the crude phenolic and terpenoid extracts Compared with12.00 and11.67 nymph/ Female in the control treatment respectively

Ganoderma terpenoid extract exhibited anti-plasmodial activity by a mechanism involving reduction in erythrocyte and hepatic lipids in Plasmodium berghei infected mice

Bioactive components of Ganoderma lucidum has recently gained intense research attention due to their acclaimed nutritional and medicinal properties. Thus, the terpenoid extract from the fruit bodies of G. lucidum (GT) was evaluated for activity against Plasmodium berghei in mice in two separate experiments. In addition, the effects of the extract on erythrocyte and hepatic lipids as well as liver HMG-CoA reductase activity before and after the treatments were also assessed. Mice with established infection were administered 100 and 250 mg/kg/day GT alone and in combination with chloroquine (CQ), in either case two separate controls designated: CQ (30 mg/kg chloroquine) and INF-CTR (1 mL DMSO) were also included. Treatment was administered orally for 12 days and parasitemia determined every three days. Percentage survival was significantly increased to 87% from 66% due to combination of GT100 with CQ compared to GT100 alone and to 75% from 62% when GT250 was administered with CQ compared to GT250 alone. Erythrocyte triglycerides, total cholesterol (TC), LDL and phospholipids contents were significantly lower in GT + CQ-treated mice compared to CQ alone and INF-CTR. Similarly, hepatic TC and phospholipid levels were significantly lower in the GT + CQ-treated mice compared to CQ alone and INF-CTR and HMG-CoA reductase activity in the liver was significantly inhibited due to administration of GT + CQ. Data from this study suggest that the anti-plasmodial action of GT could involve mechanisms associated with its hypolipidemic activity. It was also demonstrated that chloroquine, when administered in combination with GT, potentiates its curative effect in P. berghei-infected mice.

A new method for microwave assisted ethanolic extraction of Mentha rotundifolia bioactive terpenoids.

A new microwave-assisted extraction (MAE) method using ethanol as solvent has been optimized by means of a Box-Behnken experimental design for the enhanced extraction of bioactive terpenoids from Mentha rotundifolia leaves; 100°C, 5min, 1.125g dry sample: 10mL solvent and a single extraction cycle were selected as optimal conditions. Improved performance of MAE method in terms of extraction yield and/or reproducibility over conventional solid-liquid extraction and ultrasound assisted extraction was also previously assessed. A comprehensive characterization of MAE extracts was carried out by GC-MS. A total of 46 compounds, mostly terpenoids, were identified; piperitenone oxide and piperitenone were the major compounds determined. Several neophytadiene isomers were also detected for the first time in MAE extracts. Different procedures (solid-phase extraction and activated charcoal (AC) treatment) were also evaluated for clean-up of MAE extracts, with AC providing the highest enrichment in bioactive terpenoids. Finally, the MAE method here developed is shown as a green, fast, efficient and reproducible liquid extraction methodology to obtain M. rotundifolia bioactive extracts for further application, among others, as food preservatives.

Growth inhibitory effect of phenolic extracts of Ziziphus jujuba Mill. in dengue vector Aedes aegypti(L) in parent and F1 generation.

To evaluate the lethal and emergence inhibitory effect of alkaloid, phenolic and terpenoid extracts of Ziziphus jujuba (Rhamnaceae) against Aedes aegypti (Diptera: Culicidae), and to explore the effect of the most effective fraction on developmental and biochemical parameters of the dengue vector. The fourth instar larvae of Ae. aegypti were exposed to alkaloid, phenolic and terpenoid extracts from Z. jujuba leaves to test their toxicity and emergence inhibitory effects. Phenolic extract, being the most effective was further tested against the mosquitoes for their growth inhibitory effect supported by biochemical changes in the parent and F1 generation. While the different secondary metabolite fractions i.e., alkaloid, phenolics and terpenoid caused mortality at larval and pupal stages, the LC50 value was the lowest for phenolic fraction. Further study carried out with the phenolic fraction revealed that it affected growth by decreasing adult life span, fertility and fecundity of the mosquitoes. The reduction in growth was also accompanied by decrease in carbohydrate and lipid levels. It is concluded that the phenolic extract of the leaves of Z. jujuba is a potential candidate for control of Aedes mosquitoes.

Extraction and purification of five terpenoids from olibanum by ultrahigh pressure technique and high-speed countercurrent chromatography.

Five terpenoids, including two new ones, 3,7-dioxo-tirucalla-8,24-dien-21-oic acid (2) and 3α-acetoxyl-7-oxo-tirucalla-8,24-dien-21-oic acid (3), and three known ones, boscartol A (1), 11-keto-β-boswellic acid (4), and acetyl-11-keto-boswellic acid (5), have been extracted by the ultrapressure extraction and purified by pH-zone-refining countercurrent chromatography and high-speed countercurrent chromatography from olibanum. For ultrapressure extraction, the optimal condition including 200 MPa of extraction pressure, ethyl acetate of extraction solvent, 1:20 (g/mL) of solid/liquid ratio, and 2min of extraction time were obtained. For the separation, from 1.5 g of the terpenoid extract, 220.1mg of 4, 255.5mg of 5, and 212.3mg of the mixture of 1, 2, and 3 were obtained by pH-zone-refining countercurrent chromatography under the solvent system of chloroform/ethyl acetate/methanol/water (3:1:3:2, v/v/v/v) with aqueous ammonia and trifluoroacetic acid as retention and eluter agents. The enriched mixture (210mg) was further separated by conventional high-speed countercurrent chromatography with petroleum ether/ethyl acetate/methanol/water (1:0.8:1.1:0.6, v/v/v/v), yielding 30.1mg of 1, 35.5mg of 2, 12.3mg of 3. The structures of these five terpenoids were elucidated by extensive spectroscopic methods.

Supercritical Fluid Extraction of Flavonoids and Terpenoids from Herbal compounds: Experiments and Mathematical modeling

Supercritical extraction of flavonoid compounds from Dorema aucheri Boiss., a native Iranian plant using carbon dioxide was studied. A combined theory of broken and intact cell and fluid-solid discontinuous phase equilibrium approach was utilized to explain the role of operating variables on the extraction yields. Effects of temperature, pressure, solvent flow rate and solid particle size on the extraction yield were evaluated using the mathematical modeling. The model accuracy was evaluated in two cases, with and without mass transfer coefficient as a fitting parameter. The theoretical work was also repeated for supercritical extraction of terpenoid compounds from Baizhu to further evaluation of model applicability in supercritical systems. The average absolute deviations of model predictions from experimental data for Baizhu and Dorema aucheri, using no fitting parameters, were between 3.61-8.04% and 5.38-12.62 % respectively. The same deviations using one fitting parameter showed 3.37-6.33% and 1.58-7.40 % deviation, respectively.

Pungent and Bitter, Cytotoxic and Antiviral Terpenoids from Some Bryophytes and Inedible Fungi

Most liverworts elaborate characteristic odiferous, pungent and bitter tasting compounds many of which show antimicrobial, antifungal, antiviral, allergenic contact dermatitis, cytotoxic, insecticidal, anti-HIV, superoxide anion radical release, plant growth regulatory, neurotrophic, NO production inhibitory, muscle relaxant, antiobesity, piscicidal and nematocidal activities. Several inedible mushrooms produce female spider pheromones, strong antioxidant, and cytotoxic compounds. The present paper is concerned with the extraction and isolation of terpenoids from some bryophytes and inedible fungi and their pungent and bitter taste, and cytotoxic and antiviral activity.

Investigative study on the angiotensin converting enzyme (ACE) inhibiting properties of the terpenoid extract of Crataegus monogyna using in silico models

Crataegus monogyna is mainly used in the treatment of cardiac and circulatory system disorders. In vitro and clinical studies are indicative of the fact that the hydroethanolic extract of C. monogyna has angiotensin converting enzyme (ACE) inhibitory activity. This study sought to support these claims through the use of in silico modelling techniques. Possible binding conformations for β-amyrin, oleanolic acid and ursolic acid were generated using captopril, as well as enalaprilat and lisinopril, as template ligands. The ligand binding affinity (LBA) of each was calculated and the best binding conformation of each triterpene was established. Results indicate that these naturally occuring terpenes possess in silico predicted ligand binding affinities that are superior to both the small molecule captopril and the larger molecules enalaprilat and lisinopril.

Antioxidant and immunomodulatory activities of polysaccharides from the roots of Sanguisorba officinalis

The roots of Sanguisorba officinalis are used in traditional Chinese medicine for the treatment of diseases such as inflammation and internal haemorrhage. Several scientific investigations involving extraction and pharmacological studies of terpenoids and triterpenoid glycosides from this herb have been carried out. However, little is known regarding the immunomodulatory and antioxidant properties of polysaccharides from S. officinalis. Hence the polysaccharides from this herb have been investigated here. The hot water extract of S. officinalis has been fractionated using size-exclusion chromatography to obtain four polysaccharide fractions designated as SOP-1, SOP-2, SOP-3 and SOP-4. The range of molecular masses of these fractions were from 280Da to 2000kDa, and their sugar compositions consisted mainly of fructose, glucose, xylose, arabinose, and rhamnose. The antioxidant activities of the crude polysaccharide fractions were evaluated in a biological assay using Saccharomyces cerevisiae, whereas the radical scavenging activity was measured using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Analysis of the immunomodulatory activities of these polysaccharide fractions were measured by using mouse macrophages. Most of the polysaccharide fractions have stimulated the production of nitric oxide and tumour necrosis factor-α (TNF-α), and also displayed antioxidant activities. These results suggest that the roots of S. officinalis are likely to have therapeutic value for the treatment of cancer.

Use of Terpenoids as Natural Flavouring Compounds in Food Industry

Terpenoids represent the oldest known biomolecules, having been recovered from sediments as old as 2.5 billion years. Among plant secondary metabolites, they are the most abundant and diverse class of natural compounds. The diversity of terpenoids is probably a reflection of their many biological activities in nature, which has made them a widely used resource for traditional and modern human exploitation. They are usually the main constituents of essential oils of most plants offering a wide variety of pleasant scents from flowery to fruity, to woody or balsamic notes. For this reason terpenoids constitute a very important class of compounds for flavour and fragrance industries, in fact, in the US alone, the demand is forecast to grow 3.7 percent per year to $5.3 billion in 2012. The recent patents on production and extraction of terpenoids commonly used as natural flavouring compounds in food industries are reviewed in the present manuscript.

Interactive roles of terpenoid extract from the leaves of neem plant (Azadirachta indica, A. Juss) on lead induced toxicity in pregnant rabbits.

This study was designed to investigate the interactive roles of terpenoid extract from the leaves of Neem plant (Azadirachta indica, A. Juss) on lead-induced toxicity in pregnant rabbits and the resulting litters. Terpenoid was extracted from the air dried leaves of A. indica by previously reported procedure. Pregnant rabbits (does) weighing between 1.8 - 2.0 kg was randomly divided into 4 treatment groups: Group I (Control Group) received olive oil (2 ml/kg body weight per day). Group II (Lead Group) received Lead acetate solution (50 mg/kg body weight per day). Group III (Positive Control Group) received ascorbic acid (400 mg/kg body weight per day) and Group IV (Terpenoid Group) received with terpenoid extract (300 mg/kg body weight per day). Lead acetate solution (50 mg/kg body weight per day) was later administered to animals in groups III and IV. The animals were treated for 11 days starting from day 14 of gestation period of the animals. The does and the litters (young rabbits) were sacrificed 4 weeks after parturition. Blood plasma, whole blood was obtained for biochemical and lead analysis respectively. The liver, kidney, heart and lungs were removed for lead analysis. Lead concentration in the blood and the tissues was determined by atomic absorption spectroscopy (AAS). The total protein concentration in the plasma and liver homogenates were determined using biuret reaction method. The plasma albumin concentration, hepatic marker enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were also determined by standard spectrophotometric methods. The results revealed that, the terpenoid extract was able to reduce lead concentration in the blood, liver and kidney of pregnant rabbits. The levels of reduction was found to be statistically significant (p < 0.05) but the extract had no significant effect on its concentration of lead in the tissues of the litters, suggesting that the extract could not prevent inter placenta transfer of lead. Liver protein, plasma protein and albumin concentration, ALT and AST activities were not significantly different in all the groups. Conclusively, the extract was also able to reduce the lead burden in pregnant rabbits but could not produce the same effects on the litters. Key words: Pregnant rabbit, lead, toxicity, terpenoid, blood.

DETECTION OF SESQUITERPENE LACTONES IN TEN ARTEMISIA SPECIES POPULATION OF KHORASAN PROVINCES

Objective During the last three decades sesquiterpenoid lactones have emerged as one of the largest groups of plant products with over 3000 naturally occurring substances known. The genus Artemisia characteristically contains this biologically active type of secondary metabolites. With respect to increasing interest in the cytotoxic, antibiotic, phototoxic and antineoplastic activities of sesquiterpene lactones, identification of these compounds in Khorasanian Artemisia species, that some of them are endemic to Iran, can be worth while. Materials and Methods Ten species of Artemisia were collected from different parts of Khorasan Provinces of Iran. IR spectra recorded as KBr disks and in CH2Cl2 on a Unicam dp 110 spectrometer (UK). 1H-NMR (500 MHz) spectra were measured in CDCl3 using a Bruker DRX 500 spectrometer. Number of scans was 32 for all of the samples. The Herz-Hogenauer technique was applied for preparation of the terpenoid extracts. Results Our findings revealed that A. absinthium, A. ciniformis, A. fragrans, A. khorassanica, A. kopetdaghensis, A. santolina and A. sieberi had a high amount of sesquiterpene lactones. A little or no sesquiterpene lactone had been detected in other species. Differentiation of saturated and unsaturated sesquiterpene lactones was one of the advantages of 1H-NMR application in this method. Conclusion Many of tested Khorasanian Artemisia species contained high amounts of sesquiterpene lactones

The effect of terpenoid extracts from 15 pine species on the feeding behavioural sequence of the late instars of the pine processionary caterpillar Thaumetopoea pityocampa

The feeding behaviour of the pine processionary (PPC) caterpillar Thaumetopoea pityocampa Den. and Schiff. (Lepidoptera, Thaumetopoeidae) in L3-L4 stages was explored by means of laboratory arena feeding trials and natural substrates. In the bioassays, volatile extracts of 15 pine species, 8 of which are naturally growing in Greece, were incorporated. An analytical model was developed based on the principle of multinomial logit regression with five outcomes on the basis of the behavioural feeding sequence of the caterpillars. The outcomes were the five steps in which the feeding behavioural sequence was decomposed. The model's suitability (MacFadden's rho(2)=0.229, P<10(-4)) was examined when including 10 terpenes that were judged significant through a stepwise canonical discriminant analysis. The proposed model was superior to a random one and the two models resulting from the addition and subtraction of 4 terpenes to the already 10 existing compounds. The most informative model was built on the terpenes caryophyllene oxide, terpinolene, myrcene, germacrene D, eudesmol, limonene, beta-pinene, beta-caryophyllene, alpha-pinene and manoyl oxide. The background terpenes were present in the model and of particular importance. No special behavioural role, either as promoter or inhibitor could be assigned to the individual volatile metabolites, since no constant pattern among behavioural steps was observed. For instance, beta-caryophyllene while acts as promoter of attraction and trial bite it is a suppressor of partial feeding and strongly inhibits complete needle consumption. The monoterpene limonene, on the other hand, seems to be a suppressor of partial and complete feeding. The overall methodological scheme and the analytical modelling tool could be proved a suitable research protocol in unfolding the ecological role of a complex mixture of secondary metabolites. Those who develop safe practical systems can use this.