Ethyl Acetate Extract Of Roots Research Articles

In vitro Evaluation of Cytotoxicity and Antibacterial Activities of Ribwort Plantain (Plantago Lanceolata L.) Root Fractions and Phytochemical Analysis by Gas Chromatography-Mass Spectrometry.

Ribwort plantain (Plantago lanceolata L.), which belongs to the Plantaginaceae family, has been widely used as a herbal plant in traditional medicine across the globe. The present study aimed to investigate the biologically active substances of P. lanceolata root fractions, as well as the cytotoxic and antibacterial activities of extracts. The cytotoxic activity of ethyl acetate, dichloromethane, and n-butanol extracts of P. lanceolata root was evaluated by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. The P.lanceolata root extracts were also evaluated on gram-positive and negative bacteria by disc diffusion and microtiter broth dilution methods. The phytochemical content was also examined by gas chromatography-mass spectrometry. The P.lanceolata root extracts were cytotoxic; IC50 values against HCT-116 at 72 h were 168.553 μg/mL, 167.458 μg/mL, and 205.004 μg/mL for ethyl acetate, dichloromethane, and n-butanol root extracts, respectively. The dichloromethane extract of P. lanceolata root had the highest inhibitory effect against S. paratyphi (14.00±1.0 mm) at the concentration of 100 mg/mL. The minimum MIC and MBC (5 and 15 mg/mL) were observed for dichloromethane extract of P. lanceolata root against S. paratyphi. The main composition of ethyl acetate extract was 1,2-Benzenedicarboxylic acid and mono(2-ethylhexyl) ester (60.93%). The major compositions in dichloromethane and n-butanol extracts were 1,2-Benzenedicarboxylic acid, mono(2-ethylhexyl) ester (60.64%) and 2-Methyl-1-butanol (.+/-.)- (17.85%). As evidenced by the results of the present research, P. lanceolata extracts are a significant source of bioactive metabolites. Therefore, they can play a prominent role in the production of pharmaceutical materials.

In vitro and Insilico cytotoxicity activity of Aconitum Heterophyllum Phyto niosomes and its ethyl acetate root extract

Niosomes as a novel drug delivery system is a nonionic surfactant-based vesicle, have attracted much attention in pharmaceutical fields due to their excellent behavior in encapsulating both hydrophilic and hydrophobic agents. In the present study a comparative free radical scavenging and anti-proliferative potential of ethyl acetate root extract of Aconitum heterophyllum (EAEAH) and Aconitum heterophyllum roots loaded phyto-niosomes (nEAEAH) were investigated. Initially the EAEAH and nEAEAH were prepared and their antioxidant potential were studied in terms of DPPH, Super oxide, Hydroxyl radical scavenging by employing the In-vitro method. Assessment of anti-proliferative activity of these extracts was done using MCF-7 cancer cell lines (human breast cancer) by employing MTT assay and trypan blue exclusion assay. In-silico docking was performed using Auto Dock Vina software to identify the suitable antagonistic ligand which can inhibit the proteins BRCA1, BRCA2 and HER2 to exert the cytotoxic effect. In the present study a concentration dependent antioxidant, and cytotoxic activity was observed for both the extracts. IC50 values were calculated for MTT and trypan blue assay and the nEAEAH showed 41.71μg/ml, 49.36μg/ml respectively, whereas for EAEAH it was found to be 54.89 and 67.13μg/ml.

Comparative Evaluation of Biomedical Applications of Zinc Nanoparticles Synthesized by Using Withania somnifera Plant Extracts.

Green synthesis of metal nanoparticles is of great importance in the modern health care system. In this study, zinc nanoparticles (ZnONPs) were synthesized using leaf and root extracts of Withania somnifera using four different solvents. ZnONPs were characterized by UV-vis spectrophotometer with a range between 350–400 nm. Scanning electron microscope revealed spherical morphology with an overall size of 70–90 nm and XRD pattern confirmed the crystalline structure. The total flavonoids, phenolic, and alkaloid contents were significantly greater in the crude extracts as compared to ZnONPs. The highest scavenging activity was observed in ZnONPs from n-hexane and ethyl-acetate extracts of roots with IC50 values of 27.36 µg/mL and 39.44 µg/mL, respectively. ZnONPs from methanol and aqueous extracts showed significant antibacterial activity against Escherichia coli, Staphylococcus aureus, and Bacillus subtilis while none of the extracts were found to have significant antifungal activity. Maximum cytotoxic activity was observed in ZnONPs synthesized from aqueous and n-hexane root extracts with LC50 values of 9.36 µg/mL and 18.84 µg/mL, respectively. The highest antidiabetic potential was exhibited by ZnONPs from n-hexane leaf extracts, i.e., 47.67 ± 0.25%. Maximum protein kinase inhibitory potential was observed in ZnONPs of ethyl-acetate extract of roots with a bald zone of 12 mm. These results indicated that Withania somnifera-based ZnONPs showed significant biological activities compared to crude extracts. These findings can further be utilized for in-vivo analysis of nano-directed drug delivery systems.

Antimicrobial Activities of Salacia oblonga Wall Leaf and Root Extracts Against Different Bacterial Strains and Fungal Isolates.

Antibiotic resistance and the hazardous nature of synthetic drugs is threatening issue in the health sector. The alternative for this problem is to focus on plants that attribute to various compounds that exhibit therapeutic properties. Therefore, the study aims to evaluate the antimicrobial efficacy of Salacia oblonga leaf and root extracts against tested human pathogens. The S. oblonga extracts showed a significant zone of inhibition against bacteria and fungi. The leaf and root extracts of S. oblonga are prepared using low polar to high polar solvents in the Soxhlet apparatus and tested on the selected bacterial and fungal strains. Agar well diffusion and broth dilution methods evaluate antibacterial activity, antifungal activity, and Minimum Inhibitory Concentration (MIC) of extracts. Among the extracts tested, the ethyl acetate extract of root showed more antimicrobial activity against the tested bacterial and fungal strains. The most susceptible bacterial and fungal species against ethyl acetate extract are Micrococcus luteus, Mycobacterium tuberculosis, Microsporum canis, Trichophyton interdigitale, and Microsporum gypseum. The MIC for bacteria ranged from 13.0 to > 200µg/ml, whereas for fungi, the MIC ranged from 25.9 to > 200µg/ml. Ethyl acetate extract of root with 100µg/ml concentration showed 29.1mm and 28.7mm zone of inhibition against bacterial strains M. luteus and M. tuberculosis, respectively. The ethyl acetate extract of root with a 100µg/ml concentration showed 15.8, 15.2, and 15.6mm zone of inhibition against fungal isolates M. canis, T. interdigitale, and M. gypseum, respectively. The activity of root and leaf extracts increased in a concentration-dependent manner, and further, the compounds isolated from the crude extracts of leaf and root showed antimicrobial activity. Structural elucidation of isolated compounds Lambertic acid and Ferruginol was done using NMR spectroscopy. Reports indicate that Lambertic acid was isolated previously, but the isolation of hydroxy Ferruginol from S. oblonga leaf extract was reported unprecedented.

IN VITRO AND IN VIVO ANTIDIABETIC EFFECTS OF SALACIA OBLONGA WALL LEAF AND ROOT EXTRACTS

The present study was framed out to evaluate the antidiabetic effects of S. oblonga Wall leaf and root ethyl acetate and toluene extracts in vitro and in vivo. The root ethyl acetate extract found significant inhibition of alpha amylase and alpha glucosidase enzyme functions in vitro. The in vivo studies are carried out according to the guide line of CPCSEA and IAEC. The acute toxicity test of the extracts resulted in the no effect on the animals. The ethyl acetate extract of root showed the increase in body weight of animals at the end of the experiment. The blood glucose levels of diabetic rats are significantly reduced after the treatment of root ethyl acetate extract. The leaf toluene and ethyl acetate extract exhibited average antidiabetic activity. We conclude that root extract exhibited good antidiabetic activity comparing to leaf extract.

Susceptibility of Aedes albopictus Larvae to the Larvicidal Activity of Three Types of Derris elliptica Extract.

<b>Background and Objective:</b> The methanol, ethyl acetate and n-hexane extracts of <i>D. elliptica</i> root have high larvicidal activity against <i>Aedes aegypti</i> larvae, the primary vector of dengue but have not been understood their potential against <i>Ae. albopictus</i> larvae, the secondary vector of dengue that also transmits Chikungunya and Zika viruses. This <i>in vitro</i> study aims to understand the larvicidal activity of the 3 extract types of <i>D. elliptica </i>root against <i>Ae. albopictus</i> larvae. <b>Materials and Methods:</b> The tuba root extract types were obtained from the sequential extraction process with 3 steps of liquid-liquid partition as described in the previous report. Six concentrations were occupied in this experiment ranging of 0.5, 1.0, 2.0, 4.0, 10.0 and 15.0 mg L<sup>1</sup> each concentration was 5 times replicated and placed in 250 mL plastic cups. As many as 20 of 3rd instar larvae of <i>Ae. albopictus</i> were subjected in each treatment cup and larval mortality was observed after 24 and 48 hrs of exposure. <b>Results:</b> Larval mortality rates based on concentration range of 13.75-97.00 and 43,75-100%, 14.00-44.00, 34.00-90.00%, 12.00-47.00 and 28.00-88.00%, with the LC<sub>50</sub> after 24 and 48 hrs of exposure were 2.925 and 0.414, 16.184, 2.900, 15.789 and 4.380 mg L<sup>1</sup>, respectively for methanol, ethyl acetate and n-hexane extracts. <b>Conclusion:</b> The methanol, ethyl acetate and n-hexane extract of tuba root have high larvicidal activity against <i>Ae. albopictus</i> larvae. Further study on prototype formulation of larvicide and elucidation of the specific phytochemical compounds of the extracts were necessarily conducted.

IN-VITRO ANTIMICROBIAL ACTIVITY OF ETHYL ACETATE EXTRACT OF CENTAUREA BEHEN LINN

This work aims to evaluate the antimicrobial potential of ethyl acetate extracts of Centaurea Behen Linn (C. Behen) on some microorganisms. Agar well diffusion method has been used to determine the antimicrobial activities of plant extracts against Gram-positive bacteria, Gram-negative bacteria (Streptococcus mutans, Staphylococcus aureus, Salmonella bongori and Escherichia coli), and two fungus (Candida albicans and Aspergillus niger). The extracts exhibited both antibacterial and antifungal activities against tested microorganisms using standard ofloxacin, ciprofloxacin and fluconazole (10-30μg/ml). The antimicrobial activity was determined by measuring the diameter of the zone of inhibition in term of millimeter (mm). To analyze the antimicrobial activity, ethyl acetate extract of C. Behen was tested by well diffusion method against six selected strain and which shows significant inhibitory action against all the tested strain. The antimicrobial activity of ethyl acetate extract of root against all microorganisms was concentration dependent but less than standard drug. It is concluded that the antimicrobial activity showed by the plant is due to the presence of phytochemicals. For future studies, phytochemcials responsible for these activities can be isolated and modified for pharmacological purpose.

Phytochemical and biological attributes of Bauhinia variegata L. (Caesalpiniaceae).

Bauhinia variegata plant is a very popular and traditionally potent ethnomedicine. Therefore, it is need of hour to study ameliorative characteristics of B. variegata for novel secondary metabolites. The current study was designed to explore antiproliferative potential of B. variegata due to scant reports on this aspect. Extracts of various parts (flowers, leaves, bark, stem, and roots) were prepared by successive maceration using organic solvents in increasing order of polarity (n-hexane, ethyl acetate, methanol, and water). The determination of polyphenolic contents was done by using colorimetric methods while antioxidant potential was measured using reducing power assay. Brine shrimp lethality assay was performed for determining preliminary cytotoxicity and antiproliferative activity against breast cancer MCF-7 cell line using MTT protocols. Moreover, antimicrobial activities were detected by using disc diffusion assay. The alpha-amylase assay was performed to monitor the antidiabetic potential of the plant. In case of phytochemical analysis methanolic extract of leaves and bark showed highest phenolic and flavonoids contents. n-Hexane and ethyl acetate extracts of stem and roots exhibited more than 90% mortality with LD50 ranges between 1-25 µg/mL when studied by brine shrimp lethality assay. n-Hexane and ethyl acetate extracts of roots and stem also showed antiproliferative activity against human breast cancer MCF-7 cell line with IC50 values ranges between 12.10-14.20 µg/mL. Most of the extracts displayed moderately high antibacterial and antifungal activities. The n-hexane extract of roots showed antidiabetic activity with 60.80 ± 0.20% inhibition of alpha-amylase. In sum, these preliminary results will be useful for further compound isolation from selected plant parts for the discovery of antibacterial, antidiabetic, and anticancer lead candidates.

Antimicrobial Activity of Leaf, Stem and Root Extracts of Hemidesmus indicus Against Aquatic Pathogens

The present study aimed to assess the antimicrobial activity of Hemidesmus indicus against aquatic pathogens. Hemidesmus indicus is a twining shrub with lactiferous branches that produces numerous bioactive compounds. Plant parts are commonly used in traditional medicine to treat a variety of diseases. A comparative evaluation of the antimicrobial activity of Hexane, Ethyl acetate, Ethanol and water extracts of leaf, stem ad root of the Hemidesmus indicus was tested against the aquatic pathogens including bacteria and fungi by standard agar well diffusion method. The results indicated that all the plant parts exhibited varying degrees of antimicrobial properties. However, among the selected parts the highest antimicrobial property was shown by root against the bacteria Streptococcus agalactiae (16 mm), Streptococcus iniae (12 mm), Aeromonas hydrophila (20 mm), Vibrio cholera (21 mm), Pseudomonas aeruginosa (19 mm), and the fungus Aspergillus niger (19 mm) and Candida albicans (15 mm). Whereas the ethyl acetate fractions of the leaf showed maximum inhibitory activity against aquatic pathogens next to the root and the aqueous extract of the stem exhibited strong inhibition against Vibrio cholera. The study confirmed that the ethyl acetate extract of the Hemidesmus indicus root has high antibacterial activity against fish pathogens meanwhile the ethanol extract exhibited high antifungal activity. The present investigation confirmed the efficacy of Hemidesmus indicus as a natural alternative to synthetic antibiotics making promising contributions to the field of drug discovery and tremendous scope in the field of aquaculture.

Isolation and characterization of anti-tumor compounds from ethyl acetate extract of Rumex japonicus houtt roots and their cytotoxic effects

To isolate and characterize anti-tumor compounds through nuclear magnetic resonance (NMR) spectroscopy of ethyl acetate extract of R. japonicus Houtt root and their cytotoxic effects were evaluated in terms of measuring inhibition of growth of three cancer cell lines: human hepatoma cells (HepG2), cerebral glioma cells (U251) and ovarian carcinoma cells (SK-OV-3/DDP). The roots of R. japonicus Houtt were extracted with ethanol, and partitioned with petroleum ether, dichloromethane and ethyl acetate, respectively. The ethyl acetate extract was separated chromatographically using macroporous resin, positive and negative phase silica gel and high performance liquid chromatography. The structures of isolated compounds were elucidated by nuclear magnetic resonance (NMR) spectral data. Ten compounds were isolated form the ethyl acetate extract of R. japonicus roots. Their structures were determined as emodin (1), chrysophanol (2), aloe emodin (3), physcion (4), quercetin (5), resveratrol (6), β-sitosterol (7), 3β-acetoxy-28-hydroxyurs-12-ene (8), sucrose (9) and dibutyl phthalate (10). Among of them, compounds 1, 3, 4, 6 and 8 exhibited highly significant (p<0.05) cytotoxicities against HepG2 to a certain extent. Compounds 3-6 exhibited moderate cytotoxicities against U251 with IC50 values of 19.8, 23.5, 23.8 and 17.8 µM, respectively. Compounds 1, 2 and 7 also exhibited cytotoxicities against U251 to a certain extent with IC50 values of 48.3, 50.1 and 85.3 µM, respectively. Compounds 1-6 exhibited moderate cytotoxicities against (SK-OV-3/DDP) with IC50 values of 62.8, 64.2, 31.3, 61.0, 47.4 and 42.0 µM, respectively.

Qualitative and Quantitative Determination of Secondary Metabolites and Antioxidant Potential of Centaurea behen L. Root Extracts

Plants have been used by humans as a natural source for treatments and cures since ancient times, and medicinal herbs have gained popularity due to their widespread usage and lack of adverse effects. Plant study has grown in popularity across the world in recent years, and a large body of data has been gathered to demonstrate the enormous potential of medicinal plants employed in diverse traditional systems. Centaurea behen L (C. behen) is a South Asian root that belongs to the Astarcease family and is known as Safed Behman. C. behen is used to treat brain, heart, and liver weakness, as well as palpitation, hepatitis, melancholia, sexual debility, neurasthenia, spermatorrhoea, weariness, and stomach and bowel problems. It is also a heart tonic and is used to treat jaundice. Several sesqueterpene lactones, the guaianolides cyanraopicrin, augerin B, desacylcynaropicrin, grosshemin, and traces of a ketone closely related to solstitialin A were found in aerial sections of C. behen. The goal of this study was to assess in vitro antioxidant activity, as well as qualitative and quantitative phytochemical analyses of C. behen root gathered in the Bhopal region of Madhya Pradesh. The well-known test methodology was used to determine qualitative analysis of various phytochemical ingredients and quantitative analysis of total flavonoids. The antioxidant activity of an ethyl acetate extract of the roots was tested in vitro using conventional techniques against DPPH and nitric oxide (NO) radical scavenging assays. Phenols, flavonoids, tannins, saponins, alkaloids, and other phytochemicals were discovered by phytochemical investigation. Ethyl acetate, methanol, and aqueous root extract of C. behen contained 0.740, 0.381, and 0.465 mg/100mg of total flavonoids, respectively. The anti-DPPH and anti-NO actions of ethyl acetate extracts were concentration dependant. The plant's broad variety of phytochemicals implies that it has medicinal potential, which might be investigated in the pharmaceutical sector as well as in traditional medicine.

Ethyl acetate extract of Smilax glabra Roxb roots and its major active compound astilbin promote osteoblastogenesis in vitro by upregulating bone cell differentiation- associated genes

Objective: To investigate the osteoblastogenic activity of the ethyl acetate (EtOAc) extract of Smilax glabra Roxb roots and its major active compound astilbin. Methods: Astilbin was isolated from EtOAc extract using silica gel chromatography combined with fraction crystallization. Chemical structure of astilbin was determined by analysis of the spectroscopic data in comparison with the literature. MTT method was used to detect the toxicity. Alkaline phosphatase (ALP) activity was determined by the spectrophotometric method at 405 nm using p-nitrophenyl phosphate as a substrate. Calcium deposition was stained with alizarin red-S, distained with cetylpyridium chloride, and quantified at 562 nm. In silico model for astilbin-ALP interaction was analyzed using AutoDock 4.2.6. The changes in expression of osteoblast differentiation related genes were determined using quantitative real-time PCR. Results: Both the EtOAc extract and astilbin had no toxicity toward osteoblast MC3T3-E1 cells at 5.0, 10, 25, and 50 μg/mL. At 25 μg/ mL, they enhanced ALP activity and mineralization of osteoblasts up to 30% and 55% for the EtOAc extract and 22% and 41% for astilbin, respectively. Molecular docking analysis of astilbin-ALP interaction revealed Arg167, Asp320, His324, and His437 were key residues participating in hydrophobic interaction; meanwhile, His434 and Thr436 residues were involved in hydrogen bond formation in the active site of human tissue-nonspecific ALP. Moreover, the expression level of genes opn, col1, osx, and runx2 were up-regulated in astilbin treated samples with the fold changes as 2.2; 3.7; 4.1; 2.3, respectively at 10 μg/mL (P&lt;0.05). Conclusions: The EtOAc extract and its major compound astilbin exhibit osteoblastogenic activity by up-regulating important markers for bone cell differentiation. It could be a new and promising osteogenic agent with dual actions for therapeutic applications.

Evaluation of antibacterial and acute oral toxicity of Impatiens tinctoria A. Rich root extracts.

The high prevalence of morbidity and mortality from bacterial infections, together with the growing threat of antibacterial resistance, necessitated the development of alternative new drugs from traditional medicine. In Ethiopia, Impatiens tinctoria A. Rich has been traditionally used for the treatment of fungal infections such as ringworms that cause tinea pedis and it have also different medical values. Scientific information on its biological activity against a broad range of bacteria and safety data is scant, compared to its folklore data. In this study, we evaluated antibacterial activities and acute oral toxicity of aqueous, ethanol and ethyl acetate root extracts of Impatiens tinctoria A. Rich. Aqueous, ethanol and ethyl acetate extracts of the plant were evaluated using agar hole diffusion and agar dilution methods. Biological activities of the plant extracts were expressed as a zone of inhibition diameter, minimum inhibitory concentration (mg/ml), and minimum bactericidal concentration (mg/ml). The safety studies were performed by oral acute toxicity study according to the organization of economic cooperation and development test Guidelines 420.Gram-positive bacteria were more susceptible to the extracts compared to gram-negative bacteria, especially against S. aureus and S. epidermidis, which are commonly found in the skin. Ethyl acetate extract was more potent than ethanol and aqueous extracts. The 50% lethal dose (LD50) of tested mice was above 9600 mg/kg. This study provides a scientific basis for the antibacterial activity of the root extracts of I. tinctoria A. Rich, where, the ethyl acetate extract showed the most promising activity. Therefore, the antibacterial potential and practical non-toxicity of the study plant extracts suggested the possibility of using it for the development of antimicrobial drugs by further studying the plant in different directions.

UHPLC-MS Characterization and Biological Insights of Different Solvent Extracts of Two Achillea Species (A. aleppica and A. santolinoides) from Turkey.

In the current study, Achillea santolinoides and Achillea aleppica aeral parts and root were extracted with ethyl acetate, methanol, and water. Detailed phytochemical profiles were obtained using UHPLC-MS, yielding the identification of hydroxybenzoic and hydroxycinnamic acids, phenolic acid glycosides and sugar esters, acylquinic acids, O-glycosyl flavones and flavonols, and flavonoid aglycons, among others. The antioxidant properties and enzyme inhibitory activities of the extracts were assayed with in vitro tests. The phenolic content of the water extracts was significantly higher as compared to the ethyl acetate and methanol ones. A. aleppica aerial parts methanol extract possessed highest flavonoid content (49.18 mg rutin equivalent/g). Antioxidant properties assessment revealed that the methanol extract of A. santolinoides roots actively scavenged DPPH (54.11 mg TE/g) and ABTS radicals (112.53 mg TE/g) and possessed highest reducing potential (183.55 and 129.92 mg TE/g, for CUPRAC and FRAP, respectively). The ethyl acetate extracts of aerial parts and roots of both species showed highest inhibition against BuCHE (6.07–6.76 mg GALAE/g). The ethyl acetate extract of A. santolinoides aerial part showed highest inhibition against tyrosinase (73.00 mg KAE/g). These results showed that the tested Achillea species might represent novel phytotherapeutic avenues for the management of Alzheimer’s disease and epidermal hyperpigmentation conditions, which are both associated with oxidative stress. This paper could shed light into future potential industrial applications using the tested Achillea species.

PHYTOCHEMICAL SCREENING AND GC-MS ANALYSIS OF ROOT EXTRACTS OF PARKIA BIGLANDULOSA (WIGHT&amp;ARN.)

The current research work estimated to explore the qualitative and quantitative analysis of the major bioactive constituents of medicinally imperative plant Parkia biglandulosa in, the five solvent (petroleum ether, chloroform, ethyl acetate, methanolic and distilled water) root extracts of Parkia biglandulosa were predictable and characterized using GC-MS analysis. The explorations were accepted out in terms of five solvent extractions, intact extractive values, qualitative analysis, quantitative estimation and characterization of phytochemicals. The percentage value of yield extraction in petroleum ether extract was 34.74%, chloroform extract was 24.2%,ethyl acetate extract was 25%, methanol was 50% and distilled water was 54%. The preliminary phytochemical analysis showed the presence of alkaloids, flavonoids, terpenoids &amp; phenolic compounds. The total metabolites content of five solvent extracts were compared with standards. The GC-MS analysis reveals the presence of various metabolites in three solvent(petroleum ether, ethyl acetate, methanol) extracts. It signifies that results revealed the presence of various bioactive metabolites which could be demoralized for their credible solicitations for therapeutic resolves.

Larvicidal Activity of Ethyl Acetate Extract of Derris elliptica Root against the Third-Instar Larvae of Cypermethrin-Resistant Aedes aegypti Offspring.

Background:Derris elliptica extracts have a high larvicidal potential against the laboratory strain of Aedes aegypti larvae, but the effect on offspring larvae of pyrethroid-resistant strains of the species is lack understood. This study aimed to determine the larvicidal activity of the ethyl acetate extract of tuba root against the third-instar larvae of the Cypermethrin-resistant Ae. aegypti offspring.Methods:The experimental study occupied four levels of ethyl acetate extract of D. elliptica namely 10, 25, 50, and 100 ppm, and each level was four times replicated. As many as twenty of healthy third-instar larvae, offspring of Cypermethrin-resistant Ae. aegypti were subjected to each experiment group. Larval mortality rate and lethal concentration 50% subject (LC50) were calculated after 24 and 48 hours of exposure time.Results:Mortality of larvae increased directly proportional to the increase of extract concentration. Larval mortality rates after 24 and 48 hours of exposure were 40–67.5% and 62.5–97.5%, and LC50 were 34.945 and 6.461ppm, respectively.Conclusion:The ethyl acetate extract of D. elliptica has the high effectiveness larvicidal potential against the third-instar larvae, offspring of the Cypermethrin-resistant Ae. aegypti. Isolation of the specific compound is necessarily done to obtain the active ingredient for larvicide formulation.

In vitro antifungal activity, phytochemical screening and thin layer chromatography profiling of Impatiens tinctoria A. Rich root extracts

Background: The emergence of microorganisms’ new resistance mechanism, the emerging and re-emerging of new infectious disease and side effects of antimicrobials threatening our ability to treat infectious diseases. Therefore, researches for the innovation of new drugs are needed through identification of the best candidate like plants which are used in traditional medicine. In Ethiopia, women dye their palms and nails by a root paste of the study plant (Impatiens tinctoria A. Rich) as a beauty treatment, to toughen the skin and to control fungal infections.Objectives: To investigate the antifungal activities, phytochemical screening and Thin Layer Chromatography profiling of aqueous, ethanol and ethyl acetate root extracts of Impatiens tinctoria A. Rich.Methods: The roots of Impatiens tinctoria A. Rich were collected from Gurage Zone around Butajira town and processed and extracted using solvents of ethyl acetate, ethanol and water. Agar well diffusion and agar dilution methods were used for screening the antifungal activity of the extracts and for determination of minimal inhibitory concentration, respectively. The minimum fungicidal concentration of the extracts was determined and the plant extracts were subjected to phytochemical screening.Result: From the tested fungi T. rubrum and T. mentagrophytes were more susceptible. The study screened the presence of at least 7 phytochemicals (alkaloids, flavonoids, anthraquinens, terpenoids, glycosides, quinones and saponins) and 6 compounds in the roots of the study plant. Ethyl acetate extract was more potent and broader spectrum of antifungal activities than ethanol and aqueous extracts.Conclusion: The roots of the study plant showed notable antifungal potency. However, further research should be undertaken to elucidate the involved phytochemicals and compounds present in the extracts and mechanism of action which clear out the road for developing new antifungal drugs.

ISOLATION OF BIOLOGICALLY ACTIVE PLANT SECONDARY METABOLITES IN ALBIZIA CHEVALIERI USING COLUMN-CHROMATOGRAPHIC TECHNIQUES.

Plants have served human beings as natural source for treatments and therapies from ancient times to date; amongst them medicinal herbs which have gain attention because of its wide use and less side effects. Chromatographic techniques have significant role in natural products chemistry as well as contribute dramatically in the discovery of novel and innovative compounds of pharmaceutical and biomedical limportance. In the present study, phytochemical analysis of Albizia chevalieri was carried out for the three Arial parts of the plant extracted with three different solvents (methanol, ethyl acetate and n-hexane). This study also focused on step-by-step visual demonstration of fractionation and isolation of biologically active plant secondary metabolites in A.chevalieri using column-chromatographic techniques. Isolation of bioactive compounds using column-chromatographic involves: Preparation of sample; Packing of column; Pouring of sample into the column; Elution of fractions; and Analysis of each fractions using thin layer chromatography. Based-on the column-chromatographic technique, it was observed that in each fraction one band were detected. 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 phytochemical compounds suggest that A. chevalieri plant has high medicinal value and can be carefully studied to extract the natural compounds which are beneficial to human beings and that could be commercialized for higher production than using synthetic drugs with side effects. However, depending on nature of research, compounds can be further purified using high performance liquid chromatography (HPLC), and nuclear magnetic resonance (NMR)spectral analyses.

Antibacterial Components of Levisticum officinale Koch against Multidrug-resistant Mycobacterium tuberculosis

Background: A bioassay-guided fractionation technique was used to evaluate the active constituents of the perennial plant L. officinale W.D.J. Koch (Apiaceae) against multidrug resistant (MDR) Mycobacterium tuberculosis. Methods: Column chromatography was used to isolation of compounds from L. officinale and spectroscopic methods including 1D and 2D NMR (Nuclear magnetic resonance) and HRMS (high resolution mass spectrometry) were used to identification of the isolated compounds. Also, to evaluate antibacterial activity, minimum inhibitory concentration (MIC) was carried out by broth micro-dilution method. Finally, molecular docking (MD) was performed using the Schrödinger package to evaluate interactions between the active compounds and InhA protein. Results: Phytochemical analysis of the ethyl acetate extract of the plant roots led to isolation of bergapten (1), isogosferol (2), oxypeucedanin (3), oxypeucedanin hydrate (4), imperatorin (5), ferulic acid (6) and falcarindiol (7). Falcarindiol and oxypeucedanin indicated a moderate activity on MDR M. tuberculosis with MIC values of = 32 and 64 μg/mL, respectively. Antibacterial activity of falcarindiol was also observed against S. aureus and methicillin-resistant S. aureus strains with the MIC values of 7.8 and 15.6 μg/mL, respectively. The results of docking analysis showed a good affinity of oxypeucedanin (3) and falcarindiol (7) to InhA enzyme with docking score values of -7.764 and -7.703 kcal/mol, respectively. Conclusion: Finally, 7 compounds were isolated from L. officinale that compounds 2-6 report for the first time from this plant. On the basis of the molecular docking (MD) study, oxypeucedanin (3) and falcarindiol (7) as active compounds against M. tuberculosis may be proposed as potential inhibitors of 2-trans-enoyl-ACP reductase (InhA), a key enzyme involved in the biosynthesis of the mycobacterial cell wall. Moreover, antibacterial activity of falcarindiol against methicillin-resistant S. aureus (MRSA) was remarkable.

Shoot Extracts from Two Low Nodulation Mutants Significantly Reduce Nodule Number in Pea.

E107 and E132 are pea mutants that nodulate poorly. Because they have a shoot-controlled nodulation phenotype, we asked if their mutated genes were implicated in the autoregulation of nodulation (AON), a mechanism which consists of two systemic circuits, the positive CEP/CRA2 and the negative CLE/SUNN, coordinated via NIN and miR2111. We further characterized the mutants’ phenotype by studying nodule distribution and nodulation efficiency. E107 was similar to wild-type (WT) in its nodule distribution, but E132 had an extended nodulation zone with nodules forming distally on its lateral roots. Moreover, we tested whether their shoots produced a compound inhibitory to nodulation. We made ethyl-acetate extracts of roots and shoots of both mutants and WT, which we applied to rhizobia-inoculated WT seedlings and to pure rhizobial cultures. Whereas free-living bacteria were unaffected by any of the extracts, WT treated with shoot extracts from either inoculated mutant had fewer nodules than that of control. E107 and E132 shoot extracts led to a 50% and a 35% reduction in nodule number, respectively. We propose that E107 and E132 belong to a new sub-class of AON mutants, i.e., hypo-nodulators, and that their respective gene products are acting in the AON descending branch, upstream of TML signaling.