Crude Bark Extract Research Articles

Isolation, Structure Elucidation and Antimicrobial Evaluation of Natural Pentacyclic Triterpenoids and Phytochemical Investigation of Different Fractions of Ziziphus spina-christi (L.) Stem Bark Using LCHRMS Analysis.

Ziziphus spina-christi L. (ZSC-L) is a tree with thorny branches, belongs to the family Rhamnaceae and grows in the sub-tropics. The purpose of this research is to isolate and partially purify bioactive components from the crude ethanol extract of the stem bark of ZSC-L. Besides, bioassay-guided fractionation of ZSC-L stem bark was conducted using different solvents. The solvents were reutilized to minimize the production cost and environmental harm. In addition, the antimicrobial activities of the fractions were analyzed, followed by metabolic profiling using LC-HRMS. The n-butanol fraction showed the highest antimicrobial efficacy, so it was subjected to further purification. For the first time, two major compounds were isolated from the stem bark of ZSC-L and identified as lupane-type pentacyclic triterpenoids betulinic acid and betulin. Both compounds were used as antibacterial and anticancer agents and considered as a green product as the extraction procedure reduced the use of hazardous chemicals. Metabolic characterization of ZSC-L and its bioactive fractions were performed using LC-HR-ESI-MS and the results revealed the dereplication of 36 compounds belonging to different chemical classes. Flavonoids and triterpenes were the most prominent metabolite classes in the different fractions. The molecular docking results were obtained by studying the interaction of betulin and betulinic acid with antimicrobial receptors (4UYM, 1IYL, 1AJ2, 6J7L, 1AD4, 2VEG) to support the in vitro results. Our study highlights that Ziziphus spina-christi and its phytoconstituents, especially triterpenoids, act as a promising antimicrobial candidate in pharmaceutical and clinical applications.

Effect of Hybrid Type and Harvesting Season on Phytochemistry and Antibacterial Activity of Extracted Metabolites from Salix Bark.

Hundreds of different fast-growing Salix hybrids have been developed mainly for energy crops. In this paper, we studied water extracts from the bark of 15 willow hybrids and species as potential antimicrobial additives. Treatment of ground bark in water under mild conditions extracted 12–25% of the dry material. Preparative high-performance liquid chromatography is proven here as a fast and highly efficient tool in the small-scale recovery of raffinose from Salix bark crude extracts for structural elucidation. Less than half of the dissolved material was assigned by chromatographic (gas chromatography and liquid chromatography) and spectroscopic (mass spectrometry and nuclear magnetic resonance spectroscopy) techniques for low-molecular-weight compounds, including mono- and oligosaccharides (sucrose, raffinose, and stachyose) and aromatic phytochemicals (triandrin, catechin, salicin, and picein). The composition of the extracts varied greatly depending on the hybrid or species and the harvesting season. This information generated new scientific knowledge on the variation in the content and composition of the extracts between Salix hybrids and harvesting season depending on the desired molecule. The extracts showed high antibacterial activity on Staphylococcus aureus with a minimal inhibitory concentration (MIC) of 0.6–0.8 mg/mL; however, no inhibition was observed against Escherichia coli, Enterococcus faecalis, and Salmonella typhimurium. MIC of triandrin (i.e., 1.25 mg/mL) is reported for the first time. Although antibacterial triandrin and (+)-catechin were present in extracts, clear correlation between the antibacterial effect and the chemical composition was not established, which indicates that antibacterial activity of the extracts mainly originates from some not yet elucidated substances. Aquatic toxicity and mutagenicity assessments showed the safe usage of Salix water extracts as possible antibacterial additives.

Phytochemical, Cytotoxicity, and Antimycobacterial Activity Evaluation of Extracts and Compounds from the Stem Bark of Albizia coriaria Welw ex. Oliver.

Background Albizia coriaria Welw ex. Oliver (Fabaceae) is one of the plants used by herbalists in the East Africa community to prepare herbal remedies for the management of symptoms of TB. Despite its widespread use, the antimycobacterial activity of this plant was uninvestigated and there was contradicting information regarding its cytotoxicity. Methods Cytotoxicity (MTT), antimycobacterial activity (MABA), and phytochemical screening were conducted on crude extracts (hexane, chloroform, acetone, and methanol) of the stem bark of A. coriaria. Gas chromatography-mass spectrometry (GC-MS) followed by Fourier transform infrared (FTIR) spectroscopy was carried out on the acetone and methanol extracts. The binding affinities and descriptors of pharmacokinetics and toxicity of the identified compounds were predicted using computational modelling software. Results The cytotoxic concentrations of all extracts were greater than 1000 μg/mL. The minimum inhibitory concentration of both the acetone and methanol extracts was 1250.0 ± 0.0 μg/mL against M. smegmatis, whereas that against M. tuberculosis was 937.0 ± 442.0 μg/mL and 2500.0 ± 0.0 μg/mL, respectively. Hexane and chloroform extracts were not active against both strains. Alkaloids, triterpenes, flavonoids, tannins, and saponins were the predominant phytochemicals present. GC-MS analysis revealed twenty-eight and nineteen compounds in acetone and methanol extracts, respectively. Among these was hydroquinone, which was previously reported to possess antimycobacterial activity. Seven compounds identified through GC-MS analysis had better binding affinities for the mycobacterial ATPase and polyketide synthase-13 than isoniazid and rifampicin. These compounds also showed variable but promising pharmacokinetic properties with minimum toxicity. Conclusion There are phytochemicals in A. coriaria stem bark with potential antimycobacterial activity and acceptable cytotoxicity, which can be further explored and optimized for the development of novel antitubercular drugs.

Antiplasmodial Activity of Griffipavixanthone and Morelloflavone the Main Compounds from Garcinia Chromocarpa Engl. (Clusiaceae)

Ethnopharmacological relevance: The genus Garcinia is a rich source of species mostly used in traditional medicine to treat a many diseases. Bioactive constituents from this genus have been used as antimicrobial, anticancer, antimalarial, anti-inflammatory, hepatoprotective and anti-HIV. According to these properties, major compounds of this plant have been evaluated for their antiplasmodial activity. Materials and methods: The methanol extract of the G. chromocarpa stem bark was used in this study. The chromatographic fractionation of this latest led to the isolation of two major compounds, identified by NMR and MS spectroscopic techniques. The crude extract and pure compounds were compared using the in vitro evaluation of the malaria parasite growth inhibition by colorimetric method (p-LDH assay). Furthermore ABTS•+ and DPPH• scavenging activities and cytotoxicity through WST1 assay were evaluated. Results: Based on the chromatographic profile two major compounds were isolated. They were characterized by spectroscopic analysis interpretation and comparison with the data in the literature: one biflavonoid (1) and one xanthone (2). The compound 2 (Griffipavixanthone, IC50 ± S.D = 5.80±0.32 μg/mL) was found to be more active against the malaria parasite, chloroquinesensitive (3D7) strain of P. falciparum, than the compound 1 (Morelloflavone, IC50 ± S.D = 9.20±0.96 μg/mL). The antiradical activities connected to the ability to scavenge free radicals were observed for the ethanol crude extract of the stem bark, and the leave of G. chromocarpa. Conclusions: This work demonstrated for the first time the antiplasmodial activity of G. chromocarpa and its isolated major compounds. These latest are known in the genus Garcinia but, here, firstly described in G. chromocarpa Engl.

Phytochemical Analysis of Crude Plant Extracts and Laboratory Synthesized Green Nanoparticles from Acacia auriculiformis A. Cunn. ex Benth.

Recently the research and analysis of green nanoparticles (NPs) synthesized with plant extracts has been expanded significantly due to their biological activities. From the variety of agents used for nanoparticles synthesis, silver (Ag) is the most preferred due to its recommended use in medical field as best topical bactericides from ancient times. In the present study, crude bark extracts of Acacia auriculiformis A. Cunn. were used for nanoparticles synthesis and comparison of their biochemistry with crude extract was assessed by using biochemistry analytical techniques viz; UV-Visible spectrophotometer (UV-Vis), Fourier transform infra-red spectrophotometer (FTIR), Photoluminescence (PL) and Direct Light Scattering analysis ( DLS).

Antibacterial and antioxidant activities and phytochemical composition of Stereospermum kunthianum root bark

A new glycoiridoid (1) together with seven (7) known compounds were isolated from the methanol crude extract of the root bark of Stereospermum kunthianum using chromatography methods. Their structures were elucidated using HR-ESI-MS, 1 D- & 2 D-NMR spectroscopies in comparison with previous literature. The antioxidant activity was investigated by using FRAP, DPPH, ABTS and HRSA methods while the antibacterial activity was assays on Escherichia coli (ATCC25922) and Salmonella typhimurium (ATCC14028) strains. The results showed that the isolated compounds had significantly (p < 0.01) high radical scavenging (IC50) and reducing power activity. All bacteria strains showed important minimal inhibitory concentration activity against isolated compounds started at 5 mg/mL with an inhibition zone of 6 mm. Thus, the isolated compounds in S. kunthianum justify the use of the plant in traditional medicine for the treatment of various diseases in humans. These isolated compounds can be used for formulation of new drug discovery to treat infectious diseases.

28 Days Sub Acute Toxicity Studies of the Methanol Stem Bark Extract of Combretum hypopilinum Diels (Combretacea) in Rats

The aim of this study was to evaluate the safety profile of Combretum hypopilinum stem bark extracted with Methanol (70%v/v). Preliminary Phytochemical screening of the crude methanol stem bark extract was carried out, and revealed the presence of secondary metabolites such as steroids, flavonoids and alkaloids. Initial oral acute toxicity test was carried out using the Limit Dose Test to ascertain the safety of the extract in rats. Sub-acute toxicity testing was conducted by 28 days oral administration of 400 mg/kg, 800 mg/kg and 1600 mg/kg body weight to three groups of ten rats. The fourth group was administered distilled water 10 ml/kg. No major changes were observed in body weight of the animals following 28 days of daily oral administration. Biochemical parameters such as Total Protein, Total Bilirubin, Creatinine, Aspartate Transaminase (AST) and Alanine Transaminase (ALT), were found to be within normal ranges. The levels of marker enzymes in the vital organs did not show any significant changes between control and treated groups. Histopathological examination of the major vital organs (liver, brain, and kidney) revealed no significant pathological changes in the treated groups of rats. The results of the present work suggested that the methanol stem bark extract of Combretum hypopilinum is relatively safe for use at the tested doses.

Metabolomic Profiling and Antioxidant Activities of Breonadia salicina Using 1H-NMR and UPLC-QTOF-MS Analysis.

Breonadia salicina (Vahl) Hepper and J.R.I. Wood is widely used in South Africa and some other African countries for treatment of various infectious diseases such as diarrhea, fevers, cancer, diabetes and malaria. However, little is known about the active constituents associated with the biological activities. This study is aimed at exploring the metabolomics profile and antioxidant constituents of B. salicina. The chemical profiles of the leaf, stem bark and root of B. salicina were comprehensively characterized using proton nuclear magnetic resonance (1H-NMR) spectroscopy and ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). The antioxidant activities of the crude extracts, fractions and pure compounds were determined using the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging and reducing power assays. A total of 25 compounds were tentatively identified using the UPLC-QTOF-MS. Furthermore, the 1H-NMR fingerprint revealed that the different parts of plant had differences and similarities among the different crude extracts and fractions. The crude extracts and fractions of the root, stem bark and leaf showed the presence of α-glucose, β-glucose, glucose and fructose. However, catechin was not found in the stem bark crude extracts but was found in the fractions of the stem bark. Lupeol was present only in the root crude extract and fractions of the stem bark. Furthermore, 5-O-caffeoylquinic acid was identified in the methanol leaf extract and its respective fractions, while the crude extracts and fractions from the root and dichloromethane leaf revealed the presence of hexadecane. Column chromatography and preparative thin-layer chromatography were used to isolate kaempferol 3-O-(2″-O-galloyl)-glucuronide, lupeol, d-galactopyranose, bodinioside Q, 5-O-caffeoylquinic acid, sucrose, hexadecane and palmitic acid. The crude methanol stem bark showed the highest antioxidant activity in the DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging activity with an IC50 value of 41.7263 ± 7.6401 μg/mL, whereas the root crude extract had the highest reducing power activity with an IC0.5 value of 0.1481 ± 0.1441 μg/mL. Furthermore, the 1H-NMR and UPLC-QTOF-MS profiles showed the presence of hydroxycinnamic acids, polyphenols and flavonoids. According to a literature survey, these phytochemicals have been reported to display antioxidant activities. Therefore, the identified hydroxycinnamic acid (caffeic acid), polyphenol (ellagic acid) and flavonoids (catechin and (epi) gallocatechin) significantly contribute to the antioxidant activity of the different parts of plant of B. salicina. The results obtained in this study provides information about the phytochemistry and phytochemical compositions of Breonadia salicina, confirming that the species is promising in obtaining constituents with medicinal potential primarily antioxidant potential.

Histostereological Restorative and Inhibitory Effects of Crude Methanolic Bark Extract of Prunus Africanus on Testosterone-Induced Benign Prostatic Hyperplasia in Wistar Rats

Aim: To evaluate the histostereological effects of P. africanus on testosterone induced Benign Prostatic Hyperplasia (BPH) in Wistar rats.&#x0D; Place and Duration of Study: The study was carried out for six weeks in Jomo Kenyatta University of Agriculture and Technology.&#x0D; Methodology: A sample size of sixty Wistar were used as the experimental model and they were divided into two study groups of 30 rats in the restorative group and 30 rats in the inhibitory group. Each group was further categorized into 5 control and 25 experimental rats. The experimental rats were further subdivided into 5 sub-groups based on varying doses of the crude methanolic bark extract of P. africanus (0 mg, 25 mg, 50 mg, 125 mg, and 200 mg). BPH in experimental animal was induced by subcutaneous injection of testosterone propionate (7.5 mg/kg) for 10 days.&#x0D; Results: The findings of the study showed that the restorative group had a statistical significant (P&lt;0.05) ((dose dependent reduction of the prostate volume, the stromal and epithelium volume was observed. While in the inhibitory group a statistical significant (P&lt;0.05) dose-related inhibition, in the increase of the prostate volume, the stromal and epithelium volume was observed. In conclusion, the maximal restorative effect was observed to be up to 85.6% and at a dose of 200 mg, while the optimal inhibitory effects were observed to be between 66.7- 68.4% in the dose range of 100-200 mg/kg/body weight.&#x0D; Conclusion: Crude methanolic bark extract of P. africanus has both restorative.

Antioxidant, cytotoxic, and anti-venom activity of Alstonia parvifolia Merr. Bark

Objective: To evaluate antioxidant, cytotoxic, and anti-venom capacity of crude bark extracts of Alstonia parvifolia Merr. Methods: Gas chromatography-mass spectrometry (GC-MS) and energy dispersive X-ray analyses were accomplished to characterize the chemical constituents of Alstonia parvifolia. Biochemical characterization was evaluated using an inhibitory phospholipase A2 (PLA2) assay, DPPH, and cytotoxicity assays. Using the constituents listed in the GC-MS analyses, molecular docking was conducted to inspect the binding energies between the chosen compounds and selected PLA2 isoforms. Results: GC-MS analyses showed that the Alstonia parvifolia crude extract consisted predominantly of acetylmarinobufogenin (14.89%), γ-sitosterol (10.44%), 3-O-methyl-D-glucose (5.88%), 3,5-dimethoxy-4-hydroxyphenylacetic acid (5.30%), (2α,5α)-17-methoxyaspidofractinin-3-one (AFM) (4.08%), and 2,3,5,6,7,8,9-heptahydro-1-phenyl-5-(p-chlorophenylimino)-1H-benzo[e][1 , 4]thiazepine (HPT) (1.37%). The principal elemental components of Alstonia parvifolia were Ca (4.012%) and K (1.496%), as exhibited by energy dispersive X-ray examination. Alstonia parvifolia showed significant free radical scavenging ability (IC50: 0.287 mg/mL) and was non-cytotoxic to normal HDFn cells (IC50 &gt;100 μg/mL). Moreover, Alstonia parvifolia was favorably cytotoxic to MCF-7 (IC50: 4.42 μg/mL), followed by H69PR, HT-29, and THP-1, with IC50 values of 4.94, 5.07, and 6.27 μg/mL, respectively. Alstonia parvifolia also displayed notable inhibition against PLA2 activity of Naja philippinensis Taylor venom with IC50 of (15.2 ± 1.8) μg/mL. Docking and cluster analyses projected negative binding energies from AFM (-6.36 to -9.68 kcal/mol), HPT (-7.38 to -9.77 kcal/ mol), and acetylmarinobufogenin (-7.22 to -9.59 kcal/mol). These calculations were for the particular interactions of Alstonia parvifolia constituents to PLA2 homologues where the utmost affinity was detected in HPT owing to the dipole interactions with amino acid residues. Conclusions: The bark extract of Alstonia parvifolia shows great potential as an anti-venom agent due to its low cytotoxic profile, remarkable PLA2 inhibition, and docking binding energies between its bioactive constituents and PLA2 homologues.

Anti-nociceptive and Anti-inflammatory Effect of Monotheca buxifolia Leaves and Bark Extract: A Comparative Approach

Background: Inflammation plays an important role in various diseases with high prevalence within populations. Monotheca buxifolia has been long used in the folk medicine in urinary tract infections. Objective: The aim of the study was to comparatively evaluate the anti-nociceptive and anti-inflammatory activities of M. buxifolia leaves and bark along with their fractions. Methods: The anti-nociceptive activity of M. buxifolia was assessed via acetic acid induce writhing test. The anti-inflammatory activity was determined via formalin induce paw method, while in vitro anti-inflammatory and anti-nociceptive activity was carried out through COX-2 enzyme inhibition. Results: The acetic acid induce anti-nociceptive results of M. buxifolia of bark were more significant (p<0.001) then leaves crude extract of M. buxifolia at 500 mg/kg. The chloroform fraction of both leaves and bark extract also depicts significant reduction 26.5±1.0 and 23.5±0.8 in number of writhes respectively at 500 mg/kg. The anti-nociceptive effect of M. buxifolia bark in paw licking was also significant. The paw licking duration with M. buxifolia leaves crude extract was 1.9±0.01 min at 500 mg/kg dose (p<0.001) in the early phase. The chloroform fraction also showed significant (p<0.001) reduction in paw licking duration. The COX-2 inhibitory assay of leaves extract of M. buxifolia exhibited 60.0±0.5% inhibition (p<0.001) at 1000 µg/ml. The chloroform fraction of M. buxifolia leaves at 1000 µg/ml showed 42.80 ±0.3 inhibition. The bark crude extract of M. buxifolia inhibited 63.83±0.73 % (p<0.001) COX-2 at 1000 µg/ml. The chloroform fraction of M. buxifolia bark at 1000 µg/ml showed 43.83 ±0.33 % inhibition. The results were also comparable with standard drug diclofenac sodium. Conclusion: Based on promising anti-nociceptive and anti-inflammatory results, it is suggested that M. buxifolia should be a part of complementary and alternative medicine.

In vitro inhibition of multi-drug resistant Pseudomonas efflux pump by Xylopia aethiopica (Dunal) A. Rich

Global health is under constant threat due to antimicrobial drug resistance. Bacterial Infections caused by Pseudomonas aeruginosa are of importance because of their antibiotics resistance. This study aimed at evaluating the effects of extracts of Xylopia aethiopica (XA) on multidrug-resistant (MDR) Pseudomonas isolates. Fresh samples of XA leaf, stem bark and roots were collected from the botanical garden, University of Ibadan, Nigeria. Dried and pulverized samples were extracted with methanol and partitioned into n-hexane, dichloromethane and ethyl acetate. Phytochemical screening of the extracts was performed by standard methods. Antimicrobial activity and synergistic interaction were determined using microdilution and checkerboard broth dilution methods, respectively. The results revealed that crude methanol extracts of XA leaf, stem bark and root significantly (p&lt;0.05) inhibited the growth of all tested MDR Pseudomonas isolates at 10 mg/mL. At 1 mg/mL, the ethyl acetate fraction of the leaf, and dichloromethane fraction of the roots produced clear zones of inhibition of 12 – 20 mm, and minimum inhibitory concentrations (MICs) of 1 µg/mL and 0.5 mg/mL, respectively. The modulation factor (MF) of ciprofloxacin, dichloromethane fraction of XA roots and ethyl acetate fraction of XA leaf were 4, 8, and 4 on MDR isolates E01006, OAU058 and P. aeruginosa ATCC 27853, respectively. In all tested isolates, but not E01006 and E01024, the fractional MICs of ciprofloxacin/ethylacetate fraction of XA leaf extract combination was not significantly different (p&gt;0.05) compared with ciprofloxacin/verapamil combination. In conclusion, the root and leaf fractions Xylopia aethiopica that demonstrated antimicrobial activity against MDR P. aeruginosa and synergised with ciprofloxacin have the potential to rejuvenate the antimicrobial activity of ciprofloxacin in MDR P. aeruginosa.

Antiplasmodial Activity of Griffipavixanthone and Morelloflavone the Main Compounds from Garcinia Chromocarpa Engl. (Clusiaceae)

Ethnopharmacological relevanceThe genus Garcinia is a rich source of species mostly used in traditional medicine to treat a many diseases. Bioactive constituents from this genus have been used as antimicrobial, anticancer, antimalarial, anti-inflammatory, hepatoprotective and anti-HIV. The methanol stem bark extract from Garcinia chromocarpa has been fractionated using preparative HPLC and column chromatography to identify major compounds of this plant and to evaluate their antiplasmodial activity.Aim of the studyThis study was aimed to report the antiplasmodial activity of Griffipavixanthone and Morelloflavone isolated for the first time from G. chromocarpa. And the absence of cytotoxicity at the highest tested dose (100 µg/mL) have been demonstrated. This aspect could justify their use in traditional medicine.Materials and methodsChromatographic fractionation of the stem bark methanol extract led to the isolation of major compounds, identified by NMR and MS spectroscopic techniques. The crude extract and pure compounds were used for the in vitro evaluation of the malaria parasite growth inhibition by colorimetric method (p-LDH assay), of the antiradical activities and of their cytotoxicity through WST1 assay.ResultsBased on the chromatographic profile two major compounds were isolated. They were characterized by spectroscopic analysis interpretation and comparison with the data in the literature: one biflavonoid (1) and one xanthone (2). The compound 2 (Griffipavixanthone, IC50 ± S.D = 5.80±0.32 μg/mL) was found to be more active against the malaria parasite, chloroquine-sensitive (3D7) strain of P. falciparum, than the compound 1 (Morelloflavone, IC50 ± S.D = 9.20±0.96 μg/mL). The antiradical activities connected to the ability to scavenge free radicals were determined for the ethanol crude extract of the stem bark, and the leave of G. chromocarpa.ConclusionsThis work demonstrated for the first time the antiplasmodial activity of G. chromocarpa and its isolated major compounds. These latest are known in the genus Garcinia but, here, firstly described in G. chromocarpa Engl.

Antioxidant-phenolic content correlation of phenolics rich fractions from Dillenia suffruticosa wood bark

D illenia suffruticosa is one of the tropical medicinal plants that has been used traditionally for treating several tropical diseases. However, phytochemical and pharmacological investigations of D. suffruticosa wood bark have not been well-investigated. This research aimed to investigate the correlation between antioxidant properties and total phenolic content of phenolics rich extract and fractions from D . suffruticosa wood bark. The phytochemical investigation revealed the existence of phenolics and alkaloids in methanolic crude wood bark extract and all fractions. Meanwhile, the presence of flavonoids was shown in chloroform and methanol fractions. The highest total phenolic content was demonstrated by methanol fraction with 254.34 ± 16.86 mg GAE/g extract. Meanwhile, the highest total flavonoid content was interestingly displayed in chloroform fraction with 15.33 ± 0.26 mg RE/g extract. The crude extract and methanol fraction had a profound antioxidant activity with IC 50 values of less than 15.63 ppm and 8.83 ppm, respectively. A strong correlation was shown between the antioxidant activity and total phenolic contents through correlation analysis. Thus, the D. suffruticosa wood bark could be considered a potential natural source of antioxidants.

Phytochemical characterization and in vitro antibacterial activity of Xeroderris stuhlmannii (Taub.) Mendonca & E.P. Sousa bark extracts

Herbal medicine use is common in rural communities of Zimbabwe since generic drugs are expensive and not readily available. In this work, we carried out phytochemical screening of secondary metabolites and used GC MS and tandem mass spectroscopy (LC MS/MS) to characterize Xeroderris stuhlmannii (Taub.) Mendonca & E.P. Sousa bark extracts. The activity of bark extracts on six bacterial strains were tested in vitro using the disc diffusion method. Qualitative screening of secondary metabolites confirmed the presence of alkaloids, flavonoids, terpenoids, steroids and polyphenols in the crude bark extracts. Methanol extracted more secondary metabolites compared to other solvents. The minimum inhibitory concentrations (MICs) values for the crude extracts on six bacterial strains ranged from 0.23–0.80 mg/mL. Antibacterial tests showed higher potency for crude bark extracts on E. Coli (MIC, 0.232 mg/mL) and lower potency on coliform (MIC, 0.798 mg/mL). LC-MS/MS analysis of various fractions confirmed the presence of twenty-eight phytochemicals whereas, twelve phytochemicals were identified using GC-MS. Both techniques confirmed the presence of ursolic acid, roburic acid, reticuline, rotenone and p-coumaric acid glucoside in hexane and methanol extracts. Our findings demonstrate that Xeroderris stuhlmannii (Taub.) Mendonca & E.P. Sousa contain many phytochemical compounds that have antibacterial activity. Some of these compounds in the bark extract possess antioxidant, antiviral, antitumor and anti-inflammatory properties. Thus, Xeroderris stuhlmannii (Taub.) Mendonca & E.P. Sousa barks can act as a useful herbal supplement for treatment of a number of diseases in rural communities. Future work will involve isolation of bioactive phytochemicals and their full characterization using LC MS/MS as well as testing their antioxidant, antidiabetic, anticancer properties and toxicity.

Identification of 3,3\u2032-O-dimethylellagic acid and apigenin as the main antiplasmodial constituents of Endodesmia calophylloides Benth and Hymenostegia afzelii (Oliver.) Harms

BackgroundEndodesmia calophylloides and Hymenostegia afzelii belong to the Guttiferae and Caesalpiniaceae plant families with known uses in African ethno-medicine to treat malaria and several other diseases. This study aimed at identifying antiplasmodial natural products from selected crude extracts from H. afzelii and E. calophylloides and to assess their cytotoxicity.MethodsThe extracts from H. afzelii and E. calophylloides were subjected to bioassay-guided fractionation to identify antiplasmodial compounds. The hydroethanol and methanol stem bark crude extracts, fractions and isolated compounds were assessed for antiplasmodial activity against the chloroquine-sensitive 3D7 and multi-drug resistant Dd2 strains of Plasmodium falciparum using the SYBR green I fluorescence-based microdilution assay. Cytotoxicity of active extracts, fractions and compounds was determined on African green monkey normal kidney Vero and murine macrophage Raw 264.7 cell lines using the Resazurin-based viability assay.ResultsThe hydroethanolic extract of H. afzelii stem bark (HasbHE) and the methanolic extract of E. calophylloides stem bark (EcsbM) exhibited the highest potency against both Pf3D7 (EC50 values of 3.32 ± 0.15 μg/mL and 7.40 ± 0.19 μg/mL, respectively) and PfDd2 (EC50 of 3.08 ± 0.21 μg/mL and 7.48 ± 0.07 μg/mL, respectively) strains. Both extracts showed high selectivity toward Plasmodium parasites (SI > 13). The biological activity-guided fractionation led to the identification of five compounds (Compounds 1–5) from HasbHE and one compound (Compound 6) from EcsbM. Of these, Compound 1 corresponding to apigenin (EC50Pf3D7, of 19.01 ± 0.72 μM and EC50PfDd2 of 16.39 ± 0.52 μM), and Compound 6 corresponding to 3,3′-O-dimethylellagic acid (EC50Pf3D7 of 4.27 ± 0.05 μM and EC50PfDd2 of 1.36 ± 0.47 μM) displayed the highest antiplasmodial activities. Interestingly, both compounds exhibited negligible cytotoxicity against both Vero and Raw 264.7 cell lines with selectivity indices greater than 9.ConclusionsThis study led to the identification of two potent antiplasmodial natural compounds, 3,3′-O-dimethylellagic acid and apigenin that could serve as starting points for further antimalarial drug discovery.

In vitro cytotoxicity analysis of Zizyphus spina-christi stem bark extract on human cancer cell lines.

Zizyphus spina-christi (Rhamnaceae family) is an edible plant used in folk medicine. Therefore, it is of interest to report the cytotoxic effects of Z. spina-christi bark crude extract on human cell lines. Crude ethanol extract of Z. spina-christi bark was fractionated with increasing polarity (diethyl ether, chloroform, ethyl acetate and butanol fractions). The fractions were examined for their cytotoxicity against human colon cancer (HCT-116 and CACO-2), cervical cancer (HeLa and HEp-2), lung carcinoma (A-549), hepatocellular carcinoma (HepG-2), breast cancer (MCF-7) and prostate cancer (PC-3) cell lines using viability assay. Diethyl ether fraction of Z. spina-christi showed the highest cytotoxic effects among the four extracts of Z. spina-christi. The IC50 of diethyl ether fraction was 7.14, 11.2, 11.6, 15.4, 39.8, 42.2, 84.2 and 153.8 µg/ml on HepG-2, A-549, CACO-2, HCT-116, MCF-7, PC-3, HeLa, and HEp-2 cell lines, respectively. Data shows that the diethyl ether fraction of Z. spina-christi showed effective cytotoxic effects in colon, lung and hepatocellular cancer cell lines.

ELAEOCARPUS SERRATUS L. EXHIBITS POTENTIAL ANALGESIC AND ANTIDIARRHEAL ACTIVITIES IN MICE MODEL

Background. Elaeocarpus serratus L. (Family: Elaeocarpaceae) is a tropical fruit tree, traditionally used in the treatments of poisoning, diarrhea, arthritis, and other diseases. Objectives. The current study was performed to conduct the analgesic, antidiarrheal, and hypoglycemic activity of E. serratus in mice model using methanolic bark crude extract. Methods. To assess the peripheral and central analgesic activity, the acetic acid-induced writhing and tail immersion methods were respectively used. The castor-oil mediated antidiarrheal method was used to assess the antidiarrheal activity whereas, the tail tipping technique was conducted to determine the hypoglycemic activity of the plant extract. Results. In the peripheral analgesic assay, the methanolic bark crude extract of E. serratus significantly inhibits the number of writing 69.77% (200 mg/kg) and 73.26% (400 mg/kg) respectively (p&lt;0.05) which was strongly comparable with standard NSAID drug diclofenac sodium 75.58% (p&lt;0.05). Similarly, it shown a significant tail flicking response for 30 minutes, 60 minutes and 90 minutes of central analgesic activity assay. In antidiarrheal activity assay, the E. serratus substantially reduced the number of diarrheal feces 64.26% (200 mg/kg, p&lt;0.05) and 78.57% (400 mg/kg, p&lt;0.05) which was also comparable with the positive control loperamide. The hypoglycemic activity of E. serratus extract was not convincing. Conclusions. Our investigation demonstrated the significant analgesic and antidiarrheal activities of methanolic bark extract of E. serratus (200 and 400 mg/kg) in mice model.

Preliminary Phytochemical Screening and Antimicrobial Activity of Tree Bark Extract of Eucalyptus tereticornis Sm. with Reference to Air Pollution at Indore City

Purpose: Eucalyptus tereticornis Sm is used as medicine. This is safe alternative source of drugs as well. To evaluate the scientific basis for this plant species, a comprehensive study have been made for the phytochemical analysis and antimicrobial activities of crude extracts of the stem bark along with physicochemical characters with reference to air pollution condition at different polluted areas at Indore City. Methods: The present study investigated preliminary phytochemical screening of stem bark by three different solvents petroleum ether extract, ethanolic extract and distilled water extract for three different polluted areas (Mixed Polluted Area (MPA) , Vehicular Polluted Area (VPA) and Industrial polluted Area (IPA)) and was compared by Low Polluted Area (LPA) at Indore City(M.P.) for 11 Secondary metabolites which are useful for development and growth of plants. Along with this antimicrobial activity was also studied for gram negative (Escherichia coli and Klebsiella pneumoniae) and gram positive (Staphylococcus aureus and Bacillus subtilis) bacteria at all four pollution areas in two different extract (ethanolic and distilled water ) and zone of inhibition was measure in mm. Physicochemical characters was studied at all different areas. Results: Study revealed the presence of alkaloids, Carbohydrates and Tannin at all pollution areas in all three extracts in Eucalyptus tereticornis.. It was also observed that ethanolic bark extract of E. tereticornis was more effective in inhibiting at all the polluted areas including our reference area for Staphylococcus aureus the maximum zone of inhibition was 17mm at MPA. The results indicate that the ethanolic bark extracts shows more prominent results and might be exploited as natural drug for the treatment of several infectious diseases caused by these organisms.

Template-directed preparation of three-dimensionally ordered macroporous molecularly imprinted microspheres for selective recognition and separation of quinine from cinchona extract

A kind of novel three-dimensionally ordered macroporous molecularly imprinted microspheres (3DMIMs) had been developed for selective recognition and separation of quinine based on the combination of colloidal crystal templating method and molecular imprinting technique. The 3DMIMs were derived from the spherical silica colloidal crystal involving a four-step templating process including infiltration of imprinted precursors, sealing of polyethylene glycol, initiation polymerization, as well as final removal of the colloidal crystal and imprinting template. The resulted 3DMIMs consisted of a three-dimensional, highly ordered and interconnected macroporous arrays with nanoscale thickness of polymer wall, in which numerous imprinted cavities were obtained. The kinetic and isothermal adsorption behavior of the 3DMIMs were investigated in detail. The results showed the 3DMIMs had a rapider mass transfer process, higher adsorption capacities and recognition specificity toward quinine compared with the traditional bulk imprinted polymers. In addition, the 3DMIMs exhibited obvious high permeability and low backpressure at high flow rate as column packing. Extraction experiments showed that the 3DMIMs could effectively separate quinine and its diastereoisomer, quinidine, directly from crude cinchona bark extract with a satisfactory selectivity and elution recovery.