Ethyl Acetate Extract Of Roots Research Articles

Study of the Chelating Ability of Hexane, Chloroform, Ethyl acetate and Methanolic Root Extracts from Algerian Phragmites australis Species

Introduction: Heavy metals that enter organisms can cause illness and disease. Chelation therapy is a solution that uses chelating agents to protect these organisms by high accumulation. Materials and methods: The objective of this study is to evaluate the capability of different roots extracts from reed or EDTA (as standard) to complexate iron using, ferrozine-ferrous reducing assay or Zn and Cu using the murexide-Zn or Cu reducing assay in vitro. Results: the results proved that the organic phase of the first extract and EDTA (standard chelator), have the strongest chelation activities with respective rates of 70% and 97% for iron, 55% and 56% for zinc and 47% and 88% for copper compared to the control groups considered to be 100% of the complexation. Finally, the HPLC analysis showed the presence of six organic acids, which are probably responsible for this effect. Conclusion: The hexane extract and EDTA (standard), have the strongest chelation activities.
 Keywords: Phragmites australis, Chelation, Heavy metals, Organic acids, Ferrozine, Murexide.

Cytotoxic Activity of Christia vespertilionis Root and Leaf Extracts and Fractions against Breast Cancer Cell Lines.

Christia vespertilionis, commonly known as ‘Daun Rerama’, has recently garnered attention from numerous sources in Malaysia as an alternative treatment. Its herbal decoction was believed to show anti-inflammatory and anti-cancer effects. The present study investigated the cytotoxicity of the extract of root and leaf of C. vespertilionis. The plant parts were successively extracted using the solvent maceration method. The most active extract was further fractionated to afford F1–F8. The cytotoxic effects were determined using MTT assay against human breast carcinoma cell lines (MCF-7 and MDA-MB-231). The total phenolic content (TPC) of the extracts were determined. The antioxidant properties of the extract were also studied using DPPH and β-carotene bleaching assays. The ethyl acetate root extract demonstrated selective cytotoxicity especially against MDA-MB-231 with the highest TPC and antioxidant properties compared to others (p < 0.05). The TPC and antioxidant results suggest the contribution of phenolic compounds toward its antioxidant strength leading to significant cytotoxicity. F3 showed potent cytotoxic effects while F4 showed better antioxidative strength compared to others (p < 0.05). Qualitative phytochemical screening of the most active fraction, F3, suggested the presence of flavonoids, coumarins and quinones to be responsible toward the cytotoxicity. The study showed the root extracts of C. vespertilionis to possess notable anti-breast cancer effects.

Larvicidal Activity of Neolamarckia cadamba Against the Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus

The excessive use of conventional insecticides to Culicidae leads to the emergence of mosquitoe resistant, adverse environmental and human effects. Therefore, the present study was carried out to explore the larvicidal efficacy of petroleum ether, chloroform, ethyl acetate and methanol extracts of roots, leaves, and bark of Neolamarckia cadamba against the early 4th instar larvae of Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus at concentrations ranging from 12.5 to 200 ppm under laboratory conditions. The root extracts showed the highest larvicidal potential than the leaf and bark extracts. After 24 h of exposure period, the highest larvicidal activities were observed in the methanol extract of roots with LC50 value of 43.29 and LC90 value of 202.85 ppm against An. stephensi; LC50 value of 70.82 and LC90 value of 253.73 ppm against Ae. Aegypti and LC50 value of 99.51 and LC90 value of 311.02 ppm against the Cx. Quinquefasciatus, respectively. Thus, the plant extracts of N. cadamba showed considerable mosquito larvicidal activities and can be considered for further investigation.

Effect of Prosopis farcta extracts on some complications (hematology and lipid profiles) associated with alloxan induced diabetic rats

This study aimed to investigate the effects of ethyl acetate Prosopis farcta extracts on some complications associated with diabetes in rats. The experimental rats of 1.5-2 months old that weighed 190-220 g were randomly divided into six groups of seven animals each. Body weight, serum glucose, hematological changes and lipid profiles were studied. Alloxan was used of induction diabetes in rats. 252 rats were used anddifferent doses of the ethyl acetate extracts of roots, fruits, and leaves of P. farcta were orally administered daily for 28 daysfornormal and diabetic rats and the effective dose was 200 mg/kg BW for each kind of extracts. The results showed that administration of root, leaves and fruit extracts of P. farcta caused improvement in body weight, blood glucose, red blood cells (RBCs), hemoglobin (Hb), mean corpuscular hemoglobin (MCH), hematocrit (HCT orPCV) in alloxan diabetic rats in insulin like treatment manner, with the most effective improving effect for roots extract. Regarding to the effects on total and differential white blood cell (WBC) count, administration of extracts caused decrease in the total WBC and neutrophil count. Administration of root, leaves and fruit extracts of P. farcta, showed suppression in total cholesterol (TC), triglyceride (TG), very low-density lipoprotein (VLDL-C) and low-density lipoprotein (LDL-C), and increasing in high density lipoprotein (HDL-C) of alloxan diabetic rats. In conclusion, the P. farcta extracts possesses properties of improving some health complications accompanied diabetes in rats.

Studies on the Qualitative and Quantitative Phytochemical Constituents of Albizia chevalieri's Leaf, Root and Stem bark

A plant serves as a natural source of treatments and therapies for human beings as from prehistoric times to date; amongst them medicinal herbs is the best because of its extensive use and fewer side effects. In the present study, phytochemical analysis of Albizia chevalieri was carried out as this plant may be among one of the imperative medicinal herbs for the treatment of ailments like malaria, diabetes, diarrhea, and dysentery. Phytochemical analysis was carried out for the three Arial parts of the plant extracted with three different solvents (methanol, ethyl acetate and n-hexane). Qualitative analysis showed that methanol extracted almost all the secondary metabolites in all the three Arial parts of the plant. After quantification, it was observed that methanol Root extract was having the highest phenolic contents (44.83µg/GAE/g), followed by methanol leaf extract (38.89 µg/GAE/g), and methanol stem bark extract (22.00 µg/GAE/g). Root ethyl acetate extract (8.19 µg GAE/g), stem bark ethyl acetate extract (13.49 µg GAE/g), and leaves ethyl acetate extract (23.66 µg GAE/g), while n-hexane extracts were having a very low phenolic content with; n- hexane root (0.66 µg GAE/g), n-hexane stem bark extract (2.69 µg GAE/g), and n-hexane leaves extract (7.80 µg GAE/g). The presence of high amount of phyto-constituents suggest that A.chevalieri may have high medicinal value and it can be carefully studied in order to extract the natural compounds present, which may be beneficial to human beings and that which may be commercialized for higher production than using synthetic drugs with side effects.

EFFECT OF SALACIA OBLONGA ROOT EXTRACT AGAINST CLINICAL ISOLATES STAPHYLOCOCCUS AUREUS

Objective: Salacia oblonga Wall. is an important medicinal plant belonging to the family Celastraceae. The study reports the effect of S. oblonga root extracts against clinical isolate Staphylococcus aureus &#x0D; Methods: Antibacterial activity was evaluated by agar diffusion method and assay for minimum inhibitory concentration (MIC) of extract. Further, the effect of S. oblonga extract determined by DNA fragmentation and respiratory dehydrogenase enzyme activity assays.&#x0D; Results: S. oblonga ethyl acetate root extract was evaluated for antibacterial activity towards clinical isolate S. aureus. Bacterial growth was determined in treated and control cells. Extract displayed good growth inhibition and MIC of the extract was 80 μg/ml. DNA fragmentation assay was carried out, this result has shown that treated bacterial cell has DNA damage compared to the control cell. Further, respiratory dehydrogenase enzyme activity was determined. In the treated cells, enzyme activity was low compared to the control cells.&#x0D; Conclusion: Salacia oblonga root extract inhibiting the growth of S. aureus by different modes of action.

Three new ent-abietane diterpenoids from the roots of Euphorbia fischeriana and their cytotoxicity in human tumor cell lines.

Three new ent-abietane diterpenoids, termed fischerianoids A–C (1–3), were isolated and identified from the ethyl acetate extracts of roots of the medicinally valuable plant Euphorbia fischeriana. The planar and relative structures of 1–3 were established via high-resolution electrospray ionisation mass spectrometry and one- and two-dimensional nuclear magnetic resonance spectroscopic analyses, and the absolute configuration of 1 was further established via X-ray crystallography experiment. Compounds 1–3 showed selective inhibitory potency against certain human tumor cell lines with IC50 values ranging from 8.50 ± 0.13 to 35.52 ± 0.08 μM.Graphical abstract Electronic supplementary materialThe online version of this article (10.1007/s12272-019-01151-y) contains supplementary material, which is available to authorized users.

Wormwood (Artemisia absinthium L.) as a promising nematicidal and antifungal agent: Chemical composition, comparison of extraction techniques and bioassay-guided isolation

The ethyl acetate, ethanol and water extracts of different organs (flowers, leaves, stems and roots) of wormwood (Artemisia absinthium L.) were investigated to examine nematicidal and antifungal activities in an effort to promote future agricultural applications of the wormwood and its potential as an industrial crop. The results showed that highest concentration of total thiophene content (TTC), as well as nematicidal and antifungal activities were found in ethyl acetate extract of roots. TTC and biological activities were highly correlated, indicating that thiophenes played an important role in nematicidal and antifungal activities. Furthermore, four extraction techniques (Maceration, Soxhlet extraction, Supercritical fluid extraction and Ultrasound-assisted extraction) were compared by evaluating the yield of thiophenes. Supercritical fluid extraction (SFE) appeared to be the best method for extracting thiophenes with the highest TTC and superior activities. In addition, eight thiophenes (1–8) including two new compounds (1 and 2) were isolated from the SFE extract by the bioassay-guided method. All thiophenes showed potent nematicidal activities. In particular, 1–4 (50% lethal concentration (LC50) values of 2.69, 4.17, 6.13 and 7.65 mg/L, respectively) were more effective than commercial nematicide abamectin (LC50, 9.47 mg/L). Also, these thiophenes showed antifungal activities against four soil-borne fungi at different degrees, especially for compounds 1 and 2 which exhibited noticeable antifungal effects (minimum inhibitory concentration (MIC) values ranging from 8–16 mg/L). Findings of this work suggest that wormwood root and its thiophenes could be used in nematicide and fungicide agent industry.

Antidiabetic efficacies of methanolic and ethyl acetate extracts of Aristolochia ringens (Vahl) roots: in vivo comparative studies

Antidiabetic effects of methanol and ethyl acetate extracts of roots of Aristolochia ringens (Vahl) were comparatively assessed in streptozotocin (STZ)-induced rats. Pulverized root of A. ringens was soaked with methanol, filtered, and concentrated to dryness. The resulting residue was subsequently dissolved in ethyl acetate, filtered, and concentrated to dryness to obtain ethyl acetate extract. Methanol and ethyl acetate extracts of A. ringens root yielded 20.50 g (10.25%) and 16.4 g (8.2%) respectively. Both extracts (150 mg/kg bw) and metformin (14.2 mg/kg bw) used as reference were administered orally to STZ-induced diabetic rats for 7 days. Fasting blood glucose (FBG) level was monitored and percentage reduction in glycemia computed. Hepatic concentrations of glucose and glycogen as well as alpha amylase and glucose-6-phosphate dehydrogenase activities were also evaluated. FBG level persisted in animals that were diabetic but not treated (323.20 ± 1.01 mg/dL), whereas FBG levels in treated rats following administration of metformin, ethyl acetate, and methanol extracts for 7 days were 148.55 ± 1.62, 129.08 ± 2.52, and 181.21 ± 0.44 mg/dL, resulting in percentage reduction of 80.93%, 85.44%, and 69.10% in glycemia respectively. Ethyl acetate extracts from A. ringens root caused higher reduction in hepatic glucose concentration with concomitant increase in glycogen level when compared to the methanol extract. The root extracts also stimulated the activity of glucose-6-phosphate dehydrogenase while inhibiting the activity of α-amylase in the liver of the treated groups compared to the untreated groups. Findings from this study revealed that ethyl acetate extract of Aristolochia ringens displayed strong antidiabetic potential than the methanol extract of the same plant.

Investigation of Mode of Action of Anti Bacterial Activity of Salacia Oblonga Extract Against Drug Resistant Pathogen

Abstract Salacia oblonga Wall belonging to family Celastraceae contains vital phytoconstituents and has been used since long for the treatment of diabetes, inflammation and burn wounds. S. oblonga ethyl acetate root extract was evaluated for antibacterial activity towards drug resistant pathogens Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella pneumoniae. Further 260 nm absorbing material was estimated in the control and treated cells. Interestingly 260 nm absorbance material is higher in the Staphylococcus aureus. Further the effect of the plant extract on drug resistant pathogen S. aureus was examined by scanning electron microscopy (SEM). SEM results have shown that treated bacterial cells have changed in morphology, size and reduced in number. Based on these results it can be concluded that S. oblonga extract acts on membrane of the drug resistant pathogen S. aureus.

Phytochemical Investigation and Antioxidant Activity of Root Extract of &amp;lt;i&amp;gt;Aloe gilbertii Reynolds&amp;lt;/i&amp;gt;, from Konso, Southern Ethiopia

<i>Aloe gilbertii</i> belonging to the family Aloaceae is used by local people for treatment various disease such as malaria and wound healing. The main purpose of this study was to investigate phytochemicals and antioxidant activity of the Root Extract of <i>Aloe gilbertii Reynolds</i> Phytochemical screening of the CH₂Cl₂: CH₃OH (1:1) extract revealed the presence of flavonoids, anthraquinone, Alkaloids, saponins, phenol and absence of steroids and phytosterols. The anti-oxidant potential and total antioxidant potential was determined by using reducing power determination method and phosphomolybdate method respectively. The antioxidant capacity of methanol, ethanol and ethyl acetate root extracts fractions of <i>Aloe gilbertii</i> were found to be 244.5±0.631, 241.5±0.112 and 106±1.05 mg of ascorbic acid per 10 mg of dry weight of antioxidant in the reducing power, respectively and it’s total antioxidant capacity of these extract fractions were 82.21±0.028, 88.75±0.016 and 74.85±0.043mg of ascorbic acid per 10 mg of dry weight of the crude extract. Thus, the results of the present investigation infer that this plant extracts possess potent antioxidant. Therefore it is strongly recommended to isolate the antioxidant components

Arctium lappa L. root extract induces cell death via mitochondrial-mediated caspase-dependent apoptosis in Jurkat human leukemic T cells

Arctium lappa L. is a perennial herb traditionally consumed to improve well-being. It has been widely reported for its antioxidant properties; however, very little is known for its exact mechanisms underlying the anticancer activity. This study aimed to investigate the mechanisms of anticancer action for different A. lappa root extracts. Arctium lappa root was extracted with ethanol, hexane and ethyl acetate, then examined for in vitro anticancer activity against cancerous HeLa, MCF-7, Jurkat cell lines and non-cancerous 3T3 cell lines. Induction of apoptosis was determined by cellular morphological changes, mitochondrial membrane potential (ΔΨm), caspase-3/7 activity and DNA fragmentation. The active compounds present in the most potent root extracts were identified by LC-ESI-MS. Among all the extracts, ethyl acetate root extract has the highest potency with IC50 of 102.2 ± 42.4 μg/ml, followed by ethanolic root extract in Jurkat T cells, at 24 h. None of the extracts were cytotoxic against 3T3 cells, suggesting that the extracts were selective against cancerous cells only. Both ethyl acetate and ethanolic root extracts exhibited significant morphological changes in Jurkat T cells, including the detachment from adjacent cells, appearance of apoptotic bodies and cells shrinkage. The extracts treated cells also displayed an increase in caspase-3/7 activity and alteration in mitochondrial membrane potential. Only ethyl acetate root extract at IC50 induced DNA fragmentation in Jurkat T cells. LC-ESI-MS analysis of the extract revealed the presence of 8 compounds, of which only 6 compounds with various biological activities reported. These findings suggest that the ethyl acetate extract of A. lappa had strong anticancer potential and induced intrinsic apoptosis via loss of ΔΨm and activation of caspase-3/7 This study can provide new insight to the discovery of new promising lead compound in chemopreventive and chemotherapeutic strategies.

Bioactivity-guided isolation of alkamides from a cytotoxic fraction of the ethyl acetate extract of Anacyclus pyrethrum (L.) DC. roots.

Abstract Introduction. The alcohol extract of Pellitory (Anacyclus pyrethrum) roots has been previously shown to exert anticancer activities on the Human Colorectal Cancer Cell Line (HCT) by targeting apoptosis, metastasis and cell cycle arrest. However, the nature of the cytotoxic molecules associated with this activity remains unexplored. Aims. This study aims to reinvestigate Pellitory root extract as regard to its cytotoxic activity and to proceed to a bioguided fractionation to explore its active fraction and to give new insight in their phytochemical constituents. Methods. Powdered roots were subjected to repeated extraction with Petroleum ether (Pe), Chloroform (Ch), Ethyl acetate (Ea) and Methanol (Me). Pellitory extracts were then screened for cytotoxic activity using the Brine Shrimp Lethality (BSL) bioassay. Results. Ea extract exhibited a marked cytotoxic activity, with LC50 of 249.26 μg/mL in the BSL bioassay. The remaining extracts (Pe,Ch,Me) treated groups exhibited no or low mortality in the range of tested concentrations (1-1000 µg/mL). BSL assay-guided chromatographic fractionation of Ea active Extract revealed a highly cytotoxic fraction (F11) with LC50 of 42.5 µg/mL. Multistep purifications of the active F11 fraction afforded four alkamides, namely N-isobutyldeca-2,4-dienamide or Pellitorine (I), N-propyldodeca- -2,8-dienamide (II), N-isobutyltetradeca-2,4-dienamide (III) and N-propylnona-2,5- -dienamide (IV). Conclusions. This study suggests that cytotoxic activity is localized mainly in the ethyl acetate extract (Ea) of pellitory roots. BSL assay fractionation of this active extract leads to the isolation of four alkamides, including pellitorine (I). While this isobutyl alkamide has previously shown strong cytotoxic activities against human cancer cell lines, the other compounds (II to IV) were not previously reported as cytotoxic. Subsequently, the isolated alkamides will be considered in future study as candidates for in depth in-vitro evaluation of their cytotoxicity against cancer and normal cell lines. Finally, through this study, BSL assay demonstrate again its usefulness as bench-top assay in exploring plant extracts for cytotoxic compounds.

Paeonia arietina and Paeonia kesrounansis bioactive constituents: NMR, LC-DAD-MS fingerprinting and in vitro assays

Paeonia species have been valued for their ethnomedicinal uses in various countries and received much interest among the scientific community for their therapeutic properties, including anti-microbial, anti-inflammatory, anti-cancer, nephroprotective and hepatoprotective effects. The multiple phytotherapeutical applications of Paeonia species inspired us to establish the phytochemical fingerprint and to evaluate the biological properties of ethyl acetate, methanol, and aqueous extracts from the roots and aerial parts of two Paeonia species (P. arietina G. Anderson and P. kesrounansis Thiébaut). Phytoconstituents of P. arietina and P. kesrounansis extracts were analyzed using 1D and 2D NMR and LC-DAD-ESI-MS. The total content of phenolics (TPC) and flavonoids (TFC) in the extracts was also evaluated. The antioxidant activity was profiled using DPPH, ABTS, CUPRAC, FRAP, phosphomolybdenum, and metal chelation assays. Enzyme inhibitory properties were evaluated against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), tyrosinase, α-amylase, and α-glucosidase. Phytochemical analysis of P. arietina and P. kesrounansis extracts showed the presence of galloyl esters of sugars, galloyl monoterpenes, and glycosylated flavonoids. The three solvent extracts presented different behavior in the bioassays. The highest antioxidant activity, tyrosinase and AChE inhibition were observed for the methanolic extract of the aerial parts of P. kesrounansis. In addition, the ethyl acetate extracts of the aerial parts of both plants were the most effective inhibitors of α-amylase. The highest BChE inhibition was observed for root methanolic extract of P. kesrounansis while the root ethyl acetate extract of P. arietina exerted the strongest inhibition of α-glucosidase. Methanol extract of P. kesrounansis aerial parts presented the highest TPC, while TFC was greatest in the corresponding extract of P. arietina. Our findings can be considered as a starting point for future studies to further validate the effectiveness and safety profiles of these plants in folk medicine.

Polyphenol composition and antimicrobial activity of various solvent extracts from different plant parts of Moringa oleifera

The extracts of Moringa oleifera, a traditional medicinal and culinary plant containing phytochemicals with diverse therapeutic properties were investigated. Five parts of the plant were extracted with 5 different solvents to analyze the total phenolics content (TPC) and total flavonoids content (TFC) which were then tested for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, reducing potential, and total antioxidant capacity assays. The TPC-TFC were highest in leaves. TPC in ethyl acetate extract (200 ± 10 mg/g) and TFC in aqueous extract (69 ± 3 mg/g) were the highest. The antioxidant activity was highest in the leaves when compared with other parts like roots, barks, and flowers. Many phenolic compounds were quantified using UHPLC, the flavonols and hydroxybenzoic acids were dominant in the leaves, roots, barks, flowers and seeds. Myricetin (1530 ± 10 µg/g) and quercetin (985 ± 4 µg/g) in leaves along with gentisic acid (85 ± 2 µg/g) and biochanin A (45 ± 1 µg/g) in the roots were considerably higher than other phenolic components found in other parts of the plant. The antibacterial activity of methanol, ethanol, and ethyl acetate leaves extracts showed larger zones of inhibition against Pseudomonas aeruginosa than Erwinia carotovora. Using minimum inhibitory concentration (MIC), the growth inhibition of P. aeruginosa and E. carotovora were found to be 86 ± 1% (ethyl acetate extracts of roots) and 79 ± 0.3% (ethanolic extracts of leaves), respectively. These results suggested that ethyl acetate, ethanol, and methanol extracts might provide the highest amount of potentially beneficial compounds that may be helpful in treating some diseases.

Scrophularia lucida L. as a valuable source of bioactive compounds for pharmaceutical applications: In vitro antioxidant, anti-inflammatory, enzyme inhibitory properties, in silico studies, and HPLC profiles

The genus Scrophularia has received much interest with regards to its traditional uses against eczema, psoriasis, and mastitis. Yet, the medicinal properties of some species still need to be scientifically validated. The present study was designed to investigate into the biological properties of various solvent extracts (ethyl acetate, methanol, and aqueous) of the roots and aerial parts of Scrophularia lucida based on its antioxidant, anti-inflammatory, and enzyme inhibitory activities together with phytochemical screening. Our results revealed that the solvent extracts differed in their biological effectiveness. The root ethyl acetate extract showed the highest ABTS scavenging, FRAP, CUPRAC, and inhibitory activity against AChE and α-glucosidase. The ethyl acetate extract of the aerial parts displayed the highest BChE and α-amylase inhibition and antioxidant effect in the phosphomolybdenum assay, while the methanol extracts of both parts were the most effective DPPH• scavengers and tyrosinase inhibitors. The methanol extracts of the root and aerial parts also inhibited NO production in lipopolysaccharide (LPS)-stimulated murine leukemic monocyte-macrophage cell (4.99% and 10.77%, respectively), at 31.25 μg/mL concentration. The highest TPC (34.98 mg GAE/g extract) and TFC (48.33 mg RE/g extract) were observed in the ethyl acetate extract of the root and aerial parts, respectively. The most abundant compounds in the root ethyl acetate extract were luteolin (852 μg/g extract), rosmarinic acid (522 μg/g extract), and hesperidin (394 μg/g extract) while kaempferol was most abundant in the ethyl acetate extract of the aerial parts (628 μg/g extract). In silico experiments were conducted on tyrosinase and the higher docking values were observed for rosmarinic acid and hesperidin. The present findings provide base line information which tend to support the potential use of S. lucida in the management of several chronic diseases, including Alzheimer’s disease and diabetes mellitus.

Ethyl Acetate Extract of Licorice Root (Glycyrrhiza glabra) Enhances Proliferation and Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells.

Glycyrrhiza glabra (G. glabra) has been used as a flavoring and sweetener agent, in addition to its therapeutic properties. It is rich in phytoestrogen and may prevent osteoporosis caused by estrogen deficiency; however, there is no evidence for its effects on proliferation and osteogenesis in mesenchymal stem cells. So, we were encouraged to investigate whether the ethyl acetate extract of licorice root as a source of phytoestrogen can act similar to estrogen in cell culture. Furthermore, the analysis of the licorice extract (LE) based on HPLC-DAD-ESI-MS indicated that LE comprises phytoestrogen compounds, such as glabridin and glabrene. In this study, the effects of LE on proliferation of human bone-marrow mesenchymal stem cells (hBM-MSCs) were investigated using MTT assay. In addition, its effects on the osteogenesis were evaluated using alkaline phosphatase activity (ALP), calcium deposition, and bone specific gene expression such as ALP, osteocalcin, Runx2, and BMP-2. The quantitative gene expression was studied by real-time RT-PCR. Our results showed a significant increase in proliferation in presence of LE in concentration 10-50 µg/mL. The differentiation of hBM-MSCs increased in doses of LE (10-25 µg/mL) compared to the control group. The effects of LE were similar to those of 17β-estradiol (E2) (10-8 M) and were abolished by ICI 182,780 an antagonist of estrogen receptor (ER) (10-7), indicating that the stimulatory effects of LE occur through estrogen receptor-mediated mechanism . Taking these into account, LE may be a potential candidate for prevention of osteoporosis in menopausal women.

Nematostatic activity of root extracts of banana (Musa spp.) genotypes as pre-infectional resistance mechanism against the burrowing nematode, Radopholus similis

ABSTRACTBurrowing nematode (Radapholus similis) is a major threat to banana (Musa spp.) production throughout the world. All commercial banana cultivars are lacking of R. similis resistance. The conferred resistance available in non-commercial banana genotypes opens ways to develop new hybrids with R. similis resistance by conventional or molecular-breeding techniques. Understanding the mechanism of resistance is the primary step that can provide Musa breeders and molecular biologists with the necessary principles to create nematode-resistant plants. Phenols and lignin have been ascribed as post-infectional resistant mechanism in banana. However, information on anti-nematode compounds that are presented in resistant banana plants before challenge by R. similis is limited. Therefore, the roots of some resistant/tolerant genotypes were tested for anti-nematode bioactivity by root amendment study. The root homogenates of resistant/tolerant genotypes (Yankambi km-5 (YKM-5), Pisang Lilin, Pisang Jaribuaya, Rose, Matti, Hatidat, Veneettu Kunnan, Then kunnan, Manoranjitham, Anaikomban, Agneeswar, Pisang Mas, Gros Michel, Williams, Red Banana (Mutant), Green Red and Tongat) inhibited the infectivity of R. similis on susceptible Musa cv. Grand Nain (AAA). To confirm their bio-activity, R. similis was exposed directly to the root extracts (water, methanol, ethyl acetate and n-hexane) of the resistant/tolerant genotypes. Methanol, ethyl acetate and n-hexane root extracts of the resistant/tolerant genotypes affected the mobility of R. similis. Results confirmed that roots of resistant/tolerant plants have pre-infectional nematostatic compounds to resist R. similis. Further bio-assay-guided fractionation, isolation and identification of nematostatic compound from methanol, ethyl acetate and n-hexane root extracts of YKM-5 was attempted. The compounds N´- (3,6-dichloro-2,7-bis (2-(diethylamino) ethoxy) dodecahydro − 9H-fluoren-9-ylidene) pivalohydrazide isolated from methanol root extract; methyl 9,12-dimethylheptadecanoate isolated from ethyl acetate extract and; 1-isopropyl-2-propyldisulfane isolated from n-hexane extract were found to have nematostatic effect on R. similis. Further effort to validate that the production of these compounds contributes to resistance against R. similis in Musa germplasm would yield concrete markers for nematode resistance breeding programs.

Semi-Preparative Separation of 10 Caffeoylquinic Acid Derivatives Using High Speed Counter-Current Chromatogaphy Combined with Semi-Preparative HPLC from the Roots of Burdock (Arctium lappa L.).

Burdock roots are healthy dietary supplements and a kind of famous traditional Chinese medicine, which contains large amounts of caffeoylquinic acid derivatives. However, little research has been reported on the preparative separation of these compounds from burdock roots. In the present study, a combinative method of HSCCC and semi-preparative HPLC was developed for the semi-preparative separation of caffeoylquinic acid derivatives from the burdock roots. The ethyl acetate extract of burdock roots was first fractionated by MCI macroporous resin chromatography and give three fractions (Fr. 1–3) from the elution of 40% methanol. Then, these three fractions (120 mg) were separately subjected to HSCCC for purification with the solvent system composed of petroleum ether-ethyl acetate-methanol-water at different volume ratios, and the mixtures were further purified by semi-preparative HPLC. As a result, a total of eight known caffeoylquinic acid derivatives including 3-O-caffeoylquinic acid (32.7 mg, 95.7%), 1,5-O- dicaffeoylquinic acid (4.3 mg, 97.2%), 3-O-caffeoylquinic acid methyl ester (12.1 mg, 93.2%), 1,3-O-dicaffeoylquinic acid (42.9 mg, 91.1%), 1,5-O-dicaffeoyl-3-O-(4-maloyl)-quinic acid (4.3 mg, 84.5%), 4,5-O-dicaffeoylquinic acid (5.3 mg, 95.5%), 1,5-O-dicaffeoyl-3-O-succinylquinic acid (8.7 mg, 93.4%), and 1,5-O-dicaffeoyl-4-O-succinylquinic acid (1.7 mg, 91.8%), and two new compounds were obtained. The new compounds were 1,4-O-dicaffeoyl-3-succinyl methyl ester quinic acid (14.6 mg, 96.1%) and 1,5-O-dicaffeoyl-3-O-succinyl methyl ester quinic acid (3.1 mg, 92.6%), respectively. The research indicated that the combination of HSCCC and semi-preparative HPLC is a highly efficient approach for preparative separation of the instability and bioactive caffeoylquinic acid derivatives from natural products.

Preparation and evaluation of antibacterial potential of Pithecellobium dulce root extract against Gram positive and Gram negative bacteria

In the present study hexane, benzene, ethyl acetate and ethanol extracts of Pithecellobium dulce root were prepared using soxhlet extractor. The extracts were evaluated for antibacterial activity against one Gram positive (Staphylococcus aureus) and three Gram negative (Acetobacter aceti, Acetobacter aceti, Klebsiella pneumoniae) strains. Disc diffusion method revealed promising antibacterial activity of the extracts prepared in polar solvents (ethyl acetate and ethanol) compared to non-polar solvents (hexane and benzene). Ethanolic root extract was found to be most active against Acetobacter aceti, Staphylococcus aureus, Klebsiella pneumonia and Enterobacter aerogenes bacterial strains. The zone of inhibition of ethanolic root extract against Acetobacter aceti, Staphylococcus aureus, Klebsiella pneumonia and Enterobacter aerogenes bacterial strains was 15.4, 11.0, 19.0 and 13.0 mm, respectively at 100 mg concentration. Ethyl acetate extract also exhibited good antibacterial activity against Entrobacter aerogenes, Klebsiella pneumonia and Acetobacter aceti. The zone of inhibition of ethyl acetate root extracts against Entrobacter aerogenes, Acetobacter aceti and Klebsiella pneumonia was 10.5, 18.0 and 10.0 mm, respectively. The benzene extract showed some activity against Acetobacter aceti with the zone of inhibition 10.0 mm. The antibacterial activity of Pithecellobium dulce root hexane extract was found to be negligible against all the four tested strains of bacteria. These findings suggest that ethanolic and ethyl acetate root extracts of Pithecellobium dulce has potential as effective anti-bacterial agent.