Crude Methanolic Leaf Extract Research Articles

Molecular Networking-Based Metabolome, In Vitro Antidiabetic and Anti-Inflammatory Effects of Breonadia salicina (Vahl) Hepper & J.R.I. Wood.

Breonadia salicina (Vahl) Hepper & J.R.I. Wood is widely distributed throughout Africa. It is used ethnobotanically to treat various diseases. However, the metabolic profile of the Breonadia species is not well characterized and the metabolites that are responsible for the bioactivity of this plant remain unknown. Therefore, there is a need to determine the phytochemical and bioactivity profile to identify metabolites that contribute to the antidiabetic, anti-inflammatory and antiproliferation activity, including the genotoxicity and cytotoxic effects, of Breonadia salicina. The study is aimed at exploring the metabolomic profile antidiabetic, anti-inflammatory and antiproliferation activity, as well as the genotoxicity and cytotoxicity effects, of constituents of B. salicina. The compounds in the B. salicina extract were analyzed by ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS), and the resultant data were further analyzed using a molecular networking approach. The crude stem bark and root extracts showed the highest antidiabetic activity against α-amylase at the lowest test concentration of 62.5 µg/mL, with 74.53 ± 0.74% and 79.1 ± 1.5% inhibition, respectively. However, the crude stem bark and root extracts showed the highest antidiabetic activity against α-glucosidase at the lowest test concentration of 31.3 µg/mL, with 98.20 ± 0.15% and 97.98 ± 0.22% inhibition, respectively. The crude methanol leaf extract showed a decrease in the nitrite concentration at the highest concentration of 200 µg/mL, with cell viability of 90.34 ± 2.21%, thus showing anti-inflammatory activity. No samples showed significant cytotoxic effects at a concentration of 10 µg/mL against HeLa cells. Furthermore, a molecular network of Breonadia species using UPLC-QTOF-MS with negative mode electrospray ionization showed the presence of organic oxygen compounds, lipids, benzenoids, phenylpropanoids and polyketides. These compound classes were differentially distributed in the three different plant parts, indicating the chemical differences between the stem bark, root and leaf extracts of B. salicina. Therefore, the identified compounds may contribute to the antidiabetic and anti-inflammatory activity of Breonadia salicina. The stem bark, root and leaf extracts of B. salicina yielded thirteen compounds identified for the first time in this plant, offering a promising avenue for the discovery of new lead drugs for the treatment of diabetes and inflammation. The use of molecular networking produced a detailed phytochemical overview of this Breonadia species. The results reported in this study show the importance of searching for bioactive compounds from Breonadia salicina and provide new insights into the phytochemical characterization and bioactivity of different plant parts of Breonadia salicina.

Phytochemical variation between hydrochloric and tartaric acid-derived alkaloidal extracts of Erythrina fusca Lour. leaves: A proton NMR-based approach

Erythrina fusca Lour. (Fabaceae) has been used traditionally to relieve toothache and migraine headaches, cleansing putrid ulcers, and reducing skin inflammations and irritation. Interestingly, most of the reported biological activities on the plant are attributed to its Erythrina alkaloids which are usually extracted through the acid-base method. While the selection of acids for the extraction process is crucial, there is limited information on comparing the efficacy of strong organic and inorganic acids in extracting Erythrina alkaloids, and, more broadly, other types of alkaloids. In this study, comparison between hydrochloric acid (HCl) and tartaric acid (H2T) was made to extract the Erythrina alkaloids. From the same mass (10 g) of the crude methanolic leaf extract, a significantly higher (9-fold) alkaloidal yield was obtained using H2T compared to the HCl. The alkaloidal extracts displayed different proton nuclear magnetic resonance (1H NMR) profile where identification of their phytochemical constituents was done extensively through comparison with respective data of previously isolated compounds from the Erythrina species. Forty-seven phytochemicals were identified in the HCl extract and they were mostly alkaloids. While the H2T extract not only contained alkaloids, but also flavonoids, and pterocarpans. Cumulatively, 83 phytochemicals were identified from the extracts, including 35 alkaloids, 18 flavonoids, 14 pterocarpans, 9 terpenes, 3 phenols, and 4 saponins. Based on multivariate analysis (MVDA), the alkaloids erysodine (1), erysovine (2), erysopine (3), warangalone (39), calopocarpin (53), and dolichin A (60) were found in both alkaloidal extracts. The PLS-DA model further identified the variation in the two extracts where the HCl extract was characterized by the flavonoids lupinifolin (41) and erythrisenegalone (43). Conversely, the H2T extract contained a broader range of phytochemicals, including the alkaloids glucoerysodine (21) and erythratidinone (28), the flavonoids 5,3′-dihydroxy-4′‑methoxy-5′-(3-methyl-1,3-butadienyl)-2′′,2′′dimethylpyrano-[5,6:6,7]-isoflavanone (51), 5,3′-dihydroxy-5′-(3‑hydroxy-3-methyl-1-butenyl)-4′‑methoxy-2′′,2′′dimethylpyrano-[5,6:6,7]-isoflavanone (52), and catechin (82), the pterocarpans calopocarpin (53) and sandwicensin (63), the terpenes loliolide (70), ursolic acid (80), propyloxyamyrin (83), and stigmast-3-en-4-one (73), the saponins sigmoisides A, B, and E (75, 76, and 77), and the phenol methyl gallate (81). The present findings highlight the value of targeting the extraction process for alkaloids, in this instance Erythrina alkaloids, using different type of acids. Furthermore, these compounds may be considered potential markers that were selectively extracted through the acid-base extraction process.

A potential antiviral against COVID-19 obtained from Byrsonima coccolobifolia leaves extract

In this study, we specifically focused on the crude methanolic leaf extract of Byrsonima coccolobifolia, investigating its antifungal potential against human pathogenic fungi and its antiviral activity against COVID-19. Through the use of high-performance liquid chromatography coupled with electrospray ionization ion trap tandem mass spectrometry, direct infusion electrospray ionization ion trap tandem mass spectrometry, and chromatographic dereplication procedures, we identified galloyl quinic acid derivatives, catechin derivatives, proanthocyanidins, and flavonoid glycosides. The broth dilution assay revealed that the methanolic leaf extract of B. coccolobifolia exhibits antifungal activity against Cryptococcus neoformans (IC50 = 4 μg/mL). Additionally, docking studies were conducted to elucidate the interactions between the identified compounds and the central residues at the binding site of biological targets associated with COVID-19. Furthermore, the extract demonstrated an in vitro half-maximum effective concentration (EC50 = 7 μg/mL) and exhibited significant selectivity (>90%) toward SARS-CoV-2.

QUANTITATIVE DETERMINATION OF PHYTOCHEMICAL CONSTITUENTS OF FRACTIONS OBTAINED FROM Ficus asperifolia LEAVES MIQ (MORACEAE) AND THE CHARACTERIZATION OF COMPOUNDS IDENTIFIED IN THE RESIDUAL AQUEOUS FRACTION

Ficus asperifolia (Miq), family Moraceae is popularly known as sand-paper tree that is found in marshy areas around river banks. In Nigeria, it is called kawusa by Hausa tribe, ipin by Yoruba tribe and asesa or amerenwa by Igbo tribe. This research aims to quantify secondary metabolites present in the crude methanol extract and fractions and to characterize the identified compounds in the residual aqueous fraction of Ficus asperifolia leaves. The powdered fruit was extracted using 6L of 70% methanol. The crude extract was dissolved in water and fractionated using chloroform, ethylacetate, and n-buthanol. Phytochemical screening was conducted to determine the chemical composition of crude methanol leaf extract of Ficus asperifolia and its fractions. The phytochemical screening conducted revealed the presence of saponins, tannins, flavonoids, alkaloids, steroids and cardiac glycosides. Quantitative analysis of total alkaloids, flavoniods, saponins, and cardiac glycosides was also carried out. The crude extract fractionated produced 16.5% of chloroform, 6.8% of ethylacetate, 5.9% of n-butanol and 70.8% of residual aqueous fractions. The extract was further characterized using the available spectroscopic techniques such as FT-IR, UV, and GC-MS respectively.

Phytochemical and antibacterial studies of methanolic extract and fractions of <i>Guiera senegalensis</i> leaves

The aim of the research study was to carry out phytochemical and antibacterial studies of methanolic leaves extract and its fractions of Guiera senegalensis. The plant material was collected, identified, shed dried and pulverized to fine powder using pestle and mortar. The powdered plant material was subjected to maceration using methanol to obtained crude methanol leaf extract which was then partitioned using n-hexane, chloroform ethyl acetate and n-butanol. The extract and the fractions obtained were subjected to phytochemical screening using standard procedure, to detect the presence of secondary metabolites. The antibacterial assay of the extract and its fractions against S. aureus, B. subtilus, P.mirabilis and S. typhyrium were investigated using agar well diffusion method at different concentration (100 – 12.5 mg/mL). The phytochemical screening revealed the presence of various secondary metabolites which varies in the extract and fractions. The extract and its fractions showed significant (p< 0.05) antibacterial activities against all the test isolates with the methanol extract having the highest mean zone of inhibition ranging from 2.00±1.00d-24.00±1.00* mm then followed by ethyl acetate fraction with the mean zone of inhibitions from 4.00±1.00d-22.00±1.00* mm the n-butanol fraction and the chloroform fraction had mean zone from 4.00±1.00d-18.50±1.00* mm and 4.00±1.00d-17.60±1.00d mm respectively while n-hexane fraction recorded lower mean zone of inhibition from 4.00±1.00a-15.00±1.00* The standard drug Ciprofloxacin had mean zone of inhibition from 16.20±0.00-39.0±0.00 mm. The most sensitive organism was S. aureus, while the least sensitive organism was S. typhyrium. The study has validated the ethnomedicinal claim for the use of this plant in treatment of antibacterial infections.

Exploring the Potential of Crude Methanolic Leaf Extract of Gossypium Hirsutum in Malaria Treatment, Pain Relief, and Inflammation Reduction

The increasing resistance of Plasmodium falciparum to standard anti-malarial drugs has led to a search for new compounds with antimalarial properties. Throughout history, various plants have been used for the treatment of malaria. Gossypium hirsutum, a plant commonly used in Ondo state for malaria treatment, was the focus of this study. The crude extracts of Gossypium hirsutum were evaluated for their anti-plasmodial, analgesic, and anti-inflammatory properties. To assess the anti-plasmodial effect, Plasmodium berghei was inoculated into 12 mice divided into four groups, with three mice in each group. Groups 1 and 2 were treated with doses of 200mg/kg.bw and 400mg/kg.bw of the crude plant extract, respectively. Group 3 received 5mg/kg.bw of chloroquine phosphate, while group 4 received normal saline. The analgesic effects were evaluated using the Eddy's hot plate method and the Tail flick method in mice. The anti- inflammatory effect was assessed using egg albumin-induced paw edema in rats. The crude methanolic extracts of Gossypium hirsutum demonstrated activity against Plasmodium parasites and a significant anti-inflammatory effect, suppressing paw edema by 49.7%. Animals administered with the extracts showed increased resptime to thermal pain induction in both the hot plate and tail flick methods, similar to the effect oonse bserved with the standard drug. The crude plant extract reduced parasitemia in mice by 79.2%, accompanied by a slight but statistically insignificant decrease in packed cell volume (PCV) and the weight of treated animals. Overall, the crude extract of Gossypium hirsutum exhibited effectiveness in managing malaria, along with a moderate analgesic and anti-inflammatory potential. These findings suggest that the plant contains active compounds with promising antimalarial effects, which could be further isolated and studied.

Sub-Acute Toxicological Evaluation of Methanol Leaf Extract of Nymphaea lotus Linn (Nymphaeceae) in Wistar Rats

Nymphaea lotus has been used for centuries as an astringent, aphrodisiac, sedative, analgesic, anti-inflammatory, and for the treatment of infectious diseases. This study is aimed at evaluating the subacute toxicity profile of methanol leaf extract of Nymphaea lotus in Wistar rats. Rats were administered the crude methanol leaf extract orally for 28 days at 250, 500, and 1,000 mg/kg. Weekly body weight, food, and water intake were recorded. On the 29th day, the rats were sacrificed, and their hematological and biochemical parameters were assessed, as well as histological examination of the kidney, liver, stomach, and intestine. The extract had no effect on the body weights, relative organ weights, or food and water intakes of the animals. It had no impact on hematological markers at the tested doses (hemoglobin, packed cell volume, red blood cell, white blood cell, mean corpuscular hemoglobin, mean corpuscular volume, and platelets concentration), except for alkaline phosphate, which was significantly (p≤0.01) greater at the 500 mg/kg. There was no significant effect on liver function parameters evaluated (total bilirubin, direct bilirubin, alanine aminotransferase, aminotransferase, and aspartate). Urea, creatinine and chloride levels were significantly elevated (p≤0.01 and p≤0.05) at 250 and 500 mg/kg doses, but not at the 1,000 mg/kg dose. Histopathological evaluation of the liver, kidneys, stomach, and intestine revealed no notable histological abnormalities. Based on the results, the methanol leaf extract of Nymphaea lotus appears to be generally safe when taken orally at these doses.

Augmentation of in vitro cytotoxic potentiality of crude methanolic leaf extract of Olea dioica as a pivotal anticancer source

Introduction and Aim: Tumor is the major cause of death world-wide, with an approximate 10 million fatalities in 2020, or almost one in every six deaths. The concerns with the current commercially available anticancer medications' low selectivity and persistent side effects have driven the research of safer and more effective chemotherapeutic drugs. The current work is focused on determining the cytotoxic potential of crude methanolic extract of Olea dioica leaves against various cancer cell lines viz., A549 (human lung cancer), HCT116 (human colorectal cancer), HeLa (human cervical cancer), MCF-7, and MDA-MB-231 (human breast cancer).
 
 Materials and Methods: The cells were treated with crude methanolic extract at five different concentrations viz., 25, 50, 100, 250 and 500 µg/ml to assess the cell viability.
 
 Results: The results from the assay performed for five cell lines have shown that methanolic extract of O. dioica has an exponential cytotoxicity potential on HeLa cell line followed by MCF-7 (IC50 value 81.05), HCT116 (106.86) and MDA-MB-231 with IC50 value 81.05, 106.86, 141.34 and 499.24 µg/ml respectively, whereas the A549-human lung cancer cell has shown no cytotoxic activity.
 
 Conclusion: The methanol extract of the O. dioica leaf could be recognized as a pivotal source of anticancer compounds, but furthermore research is needed to investigate the chemical components of O. dioica extracts that are mainly accountable for anticancer action and to expound more precise molecular processes of cell death.

Evaluation of the Antiseizure Activity of Endemic Plant Halfordia kendack Guillaumin and Its Main Constituent, Halfordin, on a Zebrafish Pentylenetetrazole (PTZ)-Induced Seizure Model.

Epilepsy is a neurological disease that burdens over 50 million people worldwide. Despite the considerable number of available antiseizure medications, it is estimated that around 30% of patients still do not respond to available treatment. Herbal medicines represent a promising source of new antiseizure drugs. This study aimed to identify new drug lead candidates with antiseizure activity from endemic plants of New Caledonia. The crude methanolic leaf extract of Halfordia kendack Guillaumin (Rutaceae) significantly decreased (75 μg/mL and 100 μg/mL) seizure-like behaviour compared to sodium valproate in a zebrafish pentylenetetrazole (PTZ)-induced acute seizure model. The main coumarin compound, halfordin, was subsequently isolated by liquid-liquid chromatography and subjected to locomotor, local field potential (LFP), and gene expression assays. Halfordin (20 μM) significantly decreased convulsive-like behaviour in the locomotor and LFP analysis (by 41.4% and 60%, respectively) and significantly modulated galn, and penka gene expression.

Antimicrobial Activity and Phytochemical Screening of Methanolic Leaf Extract of Vernonia amygdalina

Aims: Crude methanolic leaf extract of Vernonia amygdalina was evaluated to determine its bioactive constituents, the antimicrobial properties, measure the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against some selected bacterial organisms.
 Place and Duration of Study: The investigation was carried out at University of Maiduguri in Borno State, Nigeria. The herb obtained from Lake Chad Research Institute were identified and validated by the department of Forestry and wild life, Mohammet Lawan College of Agriculture (MOLCA) in Maiduguri, Borno State.
 Methodology: Crude methanolic leaf extraction of the plant, qualitative phytochemical screening, antimicrobial sensitivity against some disease-causing organisms, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were carried out.
 Results: Phytochemical screening of V. amygdalina leaves extract revealed the presence of tannins, saponins, terpenoids, flavonoid, carbohydrates and cardiac glycosides. The antimicrobial sensitivity shows P. aeruginosa, had the highest sensitivity with effect at all concentrations (26 mm at 1000 mg/ml), the lowest against Salmonella typhi were 9 mm at 1000 mg/ml and 7 mm at 200 mg/ml, while the highest were 17 mm at 400 mg/ml and 14 mm at 200 mg/ml. When tested at all concentrations (200-1000 mg/ml), S. aureus, S. pyogenes, B. subtilis, Corynebacteria species, and K. pneumonia showed greater sensitivity than Salmonella typhi but less sensitivity to P. aeruginosa. Based on the outcomes of the MIC and MBC results, S. aureus was found to be sensitive to the extract at 100 and 200 mg/ml, while P. aeruginosa was found to have the highest sensitivity to the extract at all concentrations (25-200 mg/ml) with the exception of 12.5 mg/ml of the extract.
 Conclusion: The study highlighted the antimicrobial effects of V. amygdalina leave extracts on some pathogens thereby verifying the traditional healer’s claim. Also, it was concluded that the extract of V. amygdalina contained pharmacologically active phytochemicals which could be responsible for the numerous medicinal properties exhibited by the plant leaf extract.

Phytochemical Profile of the Methanolic Leaf Extract of Vernonia amygdalina Del. and In Vitro Anticoccidial Effect of Its Fractions against Eimeria tenella

Phytochemical screening of the crude methanolic leaf extract of Vernonia amygdalina was carried out. The extract and its fractions were tested for in vitro anticoccidial efficacy against the unsporulated oocysts of Eimeria tenella by the oocyst sporulation inhibition model in a 96-well microtitre plate. Phytochemical analysis of the extract revealed the presence of alkaloids, tannins, saponins, glycosides, and triterpenoids. Different concentrations of the extracts, including 250 mg/ml, 125 mg/ml, 62.5 mg/ml, 31.25 mg/ml, 15.625 mg/ml, and 7.8 mg/ml, were tested against freshly prepared unsporulated oocysts of E. tenella. Triplicate assays were conducted for each concentration of the extract to screen for anticoccidial activity. The methanol extract and fractions showed activity against the oocysts of E. tenella in a dose-dependent manner. The sporulation inhibition rate was determined as effective when the result showed more than 50% inhibition at each concentration level of treatment. Only the methanol extract and its butanol fraction at the lowest treatment concentration of 7.8 mg/ml showed an inhibition rate of 55% and 68%, respectively. All treatments: methanol extract (100%), hexane fraction (100%), butanol fraction (100%), and aqueous residue fraction (58%) were active at the highest concentration of 250 mg/ ml. It is concluded that the methanol leaf extract of Vernonia amygdalina and its butanol fraction had better oocyst sporulation inhibition activity against Eimeria tenella and may further be explored for their anticoccidial properties.

EFFECT OF NEOCARYA MACROPHYLLA LEAVES EXTRACTS ON MURINE MODEL OF PAIN

Pain is the most common symptom of disease, which accompanies us from an early age. It is connected to the stimulus that it invokes and is also based on the observation of psychological interpretation of the phenomena taking place. The aim of this research is evaluation of the effect of Neocarya macrophylla leaves extracts on animal model of Pain. The pulverized leaves were extracted by maceration using 90 % methanol for 7 days and filtered to retrieve the residue known as the crude methanol leaf extract (MEL). Some part of MEL (60 g) was partitioned into fractions of different solvents with increasing polarity including chloroform (CF), ethylacetate (EF), n-butanol (BF) using liquid-liquid fractionation. Acute toxicity study was carried out to determine the LD50 of CF and it was estimated to be 565 mg/kg using Lorke’s method. Analgesic studies was carried out on extract and partitioned fractions using acetic acid-induced writhing test in mice and the results revealed that the extract and fractions of the plant has significant (p<0.05) analgesic effect with EF being the most active fraction and CF being the least active; EF demonstrated significant (p<0.05) and dose-dependent analgesic activity with 83.9, 82.5 and 73.4 % inhibition of writhes at 20, 40 and 80 mg/kg, respectively; the standard drug, piroxicam (at 10 mg/kg) had 96.5 % inhibition of writhes. Qualitative phytochemical screening on the fractions revealed the presence of some phytochemicals.Conclusively, this study indicates that the leaves of N. macrophylla contains phytochemical constituents with analgesic

Mechanisms of anti-hypertensive activity of methanol leaf extract and fractions of Persea americana Mill. (Lauraceae) in rats

Background: Persea americana has been identified to possess antihypertensive properties, but the mechanisms of these properties remain unclear. This study was thus designed to elucidate the mechanisms of anti-hypertensive activity of the extract and fractions of fresh leaf of Persea americana on hypertension induced in rats.Methods: Crude methanol leaf extract of P. americana was partitioned into n-butanol, dichloromethane, chloroform, hexane and ethyl acetate. The ethyl acetate fraction was further purified through column chromatography. Doses of the extract and column fraction were tested for contraction on rat isolated thoracic rings and for in vitro angiotensin converting enzyme inhibitory activities. The test doses were further evaluated for anti-hypertensive activities on noradrenaline- and L-NAME-induced hypertension as well as atherogenic index in hyperlipidemic rats. Effects were compared with control values via one-way analysis of variance using GraphPad® version 8.0 softwareResults: P. americana leaf methanol extract and ethyl acetate column fraction 3 reduced the contractile effect of noradrenaline significantly (p < 0.001) in the isolated thoracic aortic rings through intact endothelium-mediated mechanism(s). The test doses also exhibited in vitro angiotensin converting enzyme inhibitory activities similar to that of captopril. Additionally, ethyl acetate column fraction 3 also reduced the hypertensive effect of L-NAME- induced hypertension by 23%Conclusion: Persea americana methanol leaf extract and ethyl acetate column fraction 3 exhibited significant reductions in all the tested parameters of L-NAME and noradrenaline-induced hypertension most probably through inhibition of vascular alpha adrenoceptors, angiotensin converting enzyme and amelioration of dyslipidemia.

Molecular Docking and Molecular Dynamics Studies of Antidiabetic Phenolic Compound Isolated from Leaf Extract of Englerophytum magalismontanum (Sond.) T.D.Penn.

Englerophytum magalismontanum, a medicinal plant with ethnopharmacology use, has a dearth of information regarding its antidiabetic properties. This study evaluated the crude methanol leaf extract of E. magalismontanum and its fractions for total phenolic content, antioxidant activity, and digestive enzymes (α-amylase and α-glucosidase) inhibitory activity using standard methods. The total phenolic content (56.53 ± 1.94 mg GAE/g dry extract) and DPPH Trolox antioxidant equivalent (TAE) (1.51 ± 0.66 µg/mL) of the methanol fraction were the highest among the fractions. The IC50 values of the methanol fraction against α-amylase (10.76 ± 1.33 µg/mL) and α-glucosidase (12.25 ± 1.05 µg/mL) activities were also high. Being the most active, the methanol fraction was subjected to bio-assay guided column chromatography-based enzyme inhibition to obtain a pure compound. The phenolic compound isolated and identified as naringenin inhibited α-amylase and α-glucosidase with IC50 of 5.81 ± 2.14 µg/mL and 4.77 ± 2.99 µg/mL, respectively. This is the first study to isolate naringenin from E. magalismontanum extract. The molecular docking and molecular dynamics studies demonstrated naringenin as a promising lead compound in comparison to acarbose for the treatment of diabetes through the inhibition of α-glucosidase activity.

Isolation and characterization of some flavonoids from the leaf of Tapinanthus globiferus growing on Acacia nilotica

Tapinanthus globiferus is used ethnomedicinally for the treatment of bacterial infections, inflammation, stomach pain, ulcers among others. The aim of the study was to isolate bioactive compounds from the leaf of T. globiferus growing on Acacia nilotica. The powdered plant material was extracted with 90 % methanol using cold maceration and the resulting crude methanol leaf extract was partitioned into n-hexane, chloroform, ethylacetate and n-butanol fractions. The ethylacetate fraction was chromatographed on a silica gel, sephadex LH-20 column and preparative thin-layer chromatography. (-)-Epicatechin and Quercetin 3-O-β-D-glucopyranoside were isolated and characterized by means of physiochemical and spectroscopic (1D and 2D-NMR) analyses for the first time from T. globiferus growing on A. nilotica.
 Keywords: Tapinanthus globiferus; Flavonoids; Isolation; NMR

Phytochemical and Antifungal Studies of Tapinathus globiferus Extract and Fractions

T. globiferus growing on Tamarindus indica is a hemi-parasitic plant used in ethomomedicine to treat different ailment including fungal infections. The aim of the study was to conduct phytochemical and antifungal studies of T. globiferus growing on Tamarindus indica. The plant material was collected, identified and air dried. Powdered plant material was subjected to maceration using methanol to obtained crude methanol leaf extract which was partitioned using n-hexane, chloroform ethyl acetate and n- The extract and the fractions obtained were subjected to phytochemical evaluation sing standard protocols. The antifungal studies of the methanol extract and its fractions against Aspergillus fumugatus, Aspergillus niger, Trychophytom rubrum and Trychophyton mentagrophyte were investigated using agar well diffusion method at different concentration (100 - 1.25 mg/mL). The phytochemical screening revealed the presence of various secondary metabolites which varies in the extract and fractions. The methanol extract and its fractions showed significant (p< 0.05) antifungal activity against all the fungal isolates with the ethyl acetate fraction having the highest zone of inhibition ranging from 10-39 mm then followed by the n-butanol fraction with the mean zone of inhibitions from 10-28 mm. The methanol extract and the n-hexane fraction recorded lower zone of inhibition from 9-24 mm and 8-26 mm respectively. The standard drug ketoconazole had zone of inhibition from 6-22 mm. The most sensitive organism was A. niger while the least sensitive organisms were T. mentagraphyte and A. rubrum for the ethyl acetate fraction. The study has validated the ethnomedicinal claim for the use of this plant in treatment of the fungal infections.
 Keywords: Phytochemical; Tapinanthus globiferus; Tamarindus indica

Cytotoxic and Apoptotic Properties of the Flavonoid-rich Ethyl Acetate Fraction of the Crude Methanol Leaf Extract of Syzygium samarangense (Blume) Merr. (Myrtaceae) against MCF-7 Breast Cancer Cells

Introduction. Breast cancer is the most common cancer among women in the Philippines and about 3 in every100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer.Objectives. This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the defatted crude methanol leaf extract of Syzygium samarangense in MCF-7 breast cancer cell lines.Methods. Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess in vitro anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining.Results. The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The in vitro anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC50) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC50 of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses.Conclusion. The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract ofS. samarangense possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.

In-vivo antidiabetic and antidyslipidemic effects of methanolic leaf extract of Combretum indicum in the streptozotocin-induced diabetic rats

Background Combretum indicum (locally known as Basantilata) is a notable medicinal plant belonging to the family Combretaceae. Extracts collected from leaves of this plant have activities including antibacterial, antipyretic, and antidiarrheal activities.Objective This study was designed to evaluate the crude methanolic leaf extract of C. indicum (MLCI) to evaluate its activities in hyperglycemic and dyslipidemic rats.Materials and methods In-vivo antidiabetic and antidyslipidemic activities of the extract were studied in streptozotocin-induced diabetic rat models following the standard protocol established earlier. The rats were randomly divided into groups I–V as normal control, diabetic control, metformin, MLCI 250 mg/kg, and MLCI 500 mg/kg body weight, respectively.Results and conclusion The in-vivo studies indicated concentration-dependent and significant (P<0.05, 0.01, 0.001) reductions of elevated blood glucose, total cholesterol, triglyceride, and low-density lipoprotein-cholesterol levels in the treatment groups compared with diabetes-induced control group. Simultaneously, a significant (P<0.001) rise in high-density lipoprotein-cholesterol level was also observed in the study. The results revealed the advantageous roles of the MLCI in the management of diabetes mellitus.

Preliminary Phytochemical Screening and Antibacterial Activity of the Crude Methanol Leaf Extract of Diospyros Mespiliformis Hochst (EBENACEAE)

Diospyros mespiliformis, is an evergreen tree found mostly in the savannas of Africa. The leaves have wide applications in traditional medicine which includes whooping cough, wound dressing, pneumonia, syphilis, arthritis and skin infections. The preliminary phytochemical screening showed the presence of cardiac glycosides, terpenoids, alkaloids, saponins, tannins, anthraquinones and flavonoids. The in vitro antibacterial study of crude methanol leaf extract (CME) was analysed using disc diffusion method (DDM), with two-gram positive (Staphylococcus aureus and Streptococcus pneumonia) and two-gram negative (Escherichia coli and Pseudomonas aeruginosa) bacteria. The standard antibiotic disc used was gentamicin (5µg/disc). The CME showed highest activity against Streptococcus pneumonia (12.5±1.12mm) at 150mg/ml and the lowest activity was seen against Staphylococcus aureus (11.5±1.11mm) at 200mg/ml as compared to 31.0±0.82mm and 41.0±0.82mm of the control (standard drug) respectively. Pseudomonas aeruginosa showed resistance against the CME at all concentrations including the standard drug.

Quantitative determination and characterization of polyphenols from Cissus quadrangularis L. and dietary supplements using UHPLC-PDA-MS, LC-QToF and HPTLC

Stem and leaf of Cissus quadrangularis L. (Vitaceae), indigenous to Asia and Africa, were used for medicinal and dietary purposes with limited information about the plant's phytochemistry. Stem and leaf samples were assessed for the simultaneous determination of polyphenolic compounds (catechin, epicatechin, quercetin-3-O-β-glucopyranoside, kaempferol-3-O-β-glucoside, quercetin-3-O-β-rhamnoside, leachianol F, amurensin A, pallidol, resveratrol, and quadrangularin A), using UHPLC-PDA-MS. The validation data showed that the method is precise, specific, accurate, and linear over the range of 0.5−100 μg/mL. Reversed-phase ultra-high performance liquid chromatography (UHPLC) fingerprints of the crude methanolic stem and leaf extracts of C. quadrangularis were obtained at different wavelengths based on their λmax. Polyphenolics were characterized using both UHPLC-PDA-MS and LC-QToF analysis. From liquid chromatography quadrupole time of flight-electrospray ionization mass spectrometry (LC-QToF) spectra, over 40 components were structurally correlated, and confirmation was based on the fragmentation characteristics and also from the information available in the literature. In addition to the LC-QToF method, a simple, fast HPTLC method was developed as a visual aid for the rapid qualitative analytical tool to help establish the quality assessment of botanical raw materials and dietary supplements claiming to contain Cissus.