Fractionation Of Ethanolic Extract Research Articles

Chemico-pharmacological evaluations of the dwarf elephant ear (Colocasia affinis Schott) plant metabolites and extracts: health benefits from vegetable source.

Introduction: Colocasia affinis Schott (Family: Araceae), found in the Asian region, is a traditional root vegetable consumed by the locals and well-known as Dwarf Elephant Ear. Methods: For the pharmacological exploration of this root vegetable, four kupchan fractions (i.e. HSF, DCMSF, EASF, and AQSF) from ethanolic extract of C. affinis were employed to in vitro i.e. antioxidant, cytotoxicity, and antimicrobial and in vivo i.e. antidiarrheal and analgesic assays, followed by phytochemical screening and GC-MS protocol. Result and Discussion: In the antioxidant assay, the AQSF showed promising potential with an IC50 value of 29.4μg/mL and additionally, it exhibited the greatest overall phenolic content, measuring 57.23mg GAE/gm. of extract among other fractions. The AQSF also revealed promising cytotoxic activity in brine shrimp lethality assay with an LC50 value of 1.36μg/mL. Both AQSF and EASF exhibited substantial antimicrobial efficacy against both gram-positive and gram-negative bacteria as well as various fungus species with a remarkable zone of inhibitions compared to standards. Whereas, during both the castor oil-induced antidiarrheal and acetic acid-induced writhing assay, the DCMSF at 400mg/kg dose exhibited the highest 51.16% reduction of diarrhea and 52.33% reduction of writhing. Phytochemical screening revealed several chemical groups while GC-MS study of different fractions of dwarf elephant ear ethanolic extract revealed 48 different bioactive phytochemicals in total. Several targets such as KAS, DHFR for anti-microbial activities, GLR, URO for antioxidant activities, EGFR, BCL-2 for cytotoxicity, KOR, DOR for antidiarrheal activities and COX-2, TNF-α for analgesic activities are considered for molecular docking against identified phytocompounds and standards along with ADME/T studies to ascertain their safety, efficacy and drug likeliness profiles. Conclusion: To recapitulate, our study revealed that vegetables such as dwarf elephant ear can be considered as a prospective source of therapeutics and drug development besides their nutritive food values.

Methanol Fraction of Ethanol Extract of Dialium guineense Stem Bark May Alter the Activity of Glucose 6-phosphatase/Aminotransferases and Levels of Lipids in Tissues of Diabetic Wistar Rats

Methanol Fraction of Ethanol Extract of Dialium guineense Stem Bark May Alter the Activity of Glucose 6-phosphatase/Aminotransferases and Levels of Lipids in Tissues of Diabetic Wistar Rats

The Ethanol Extract Fractionation (Peperomia Pellucida (L.) Kunth) On Uric Acid Levels Blood Serum Of Male White Mice

Research has been carried out on the effect of fractionating the ethanol extract of the messenger herbs (Peperomia pellucida (L.) Kunth) to reduce blood serum uric acid levels in male white mice. Ordered herbs containing flavonoid compounds are thought to be used as lowering uric acid levels in the blood. The purpose of this study was to determine the effect of giving the ethanol extract fraction (Peperomia pellucida (L.) Kunth) on blood serum uric acid levels of male white mice. Simplicia extraction was carried out with 96% ethanol and extract fractionation was carried out using n-hexane, ethyl acetate, and water as solvents. The chicken liver inductor was used using experimental methods in vivo. Uric acid levels in mice were measured using a test-strip out. The results of the analysis using statistical tests on the decrease in uric acid levels in the blood were significant with a value (p 0.05) on the 14th day of the ethyl acetate fraction. This study concludes that there are compounds in the messenger herbs that have the potential to reduce uric acid levels in the blood.

Antihyperlipidemic Bioactivity Guided Isolation and Structural Elucidation of Isolated Phytoconstituents from Convolvulus pluricaulis Choisy

Background: Atherosclerosis is a crucial element in the origination of coronary heart disease (CHD). Lipid aggregation inside the coronary arteries walls lead to the occurrence of hyperlipidemia. Aim: The research was conducted for assessing the lipid lowering effect of ethanol extract and chloroform fraction from Convolvulus pluricaulis Choisy. aerial parts in Triton induced hyperlipidemic rat model. Methodology: The active fraction was obtained from antihyperlipidemic bioactivity. The phytocomponents were separated from the active fraction using flash chromatography technique resulting in the isolation of n-hexatriacontane; 9-octadecenoic acid- octyl ester; 12, 14-heptacosanedione; dodecyl-octadeca-9,12-dienoate; tetracosanyl 9-hexadecenoate; heptacosan-14-ol; stigmasta- 5, 22- dien-3 beta-ol and stigmast-5-en-3 beta-ol. The phytocomponent structures were established by spectral data analysis. Intraperitoneal (i.p.) injection of Triton WR- 1339 at a dose of 400 mg/kg body weight (b.w.) was given to the animals. The ethanol extract at a dose of 200 mg/kg b.w. and chloroform fraction at a dose of 400 mg/kg b.w. were administered orally in rats after 24 hours of the Triton administration. Results: The study resulted in dose dependent inhibition of triglycerides (p <0.01), total cholesterol (p <0.05), Low density lipoprotein level (p <0.05) while significant increased HDL (p <0.01) level. Alteration in lipid profile in Triton treated rats was restored and found more effective in the chloroform fraction probably due to the presence of phytocomponents. The study elucidated different phytocomponents isolated from the active chloroform fraction of C. pluricaulis. Conclusion: Treatment with extract and fraction manifested significant decrease in triglyceride level. HDL exerts an inhibitory action in the etiology of atherosclerosis. It is considered as a beneficial lipoprotein and its lower level leads to the greater risk of CHD. It may be concluded that the isolated components were responsible for the lipid lowering level of active fraction. GRAPHICAL ABSTRACT

Evaluation of the merit of ethanolic extract of Annona reticulata as an anti-cancer agent in human colon cancer cell lines (HCT-116)

Background Colon cancer is the fourth most common cancer type worldwide. Novel alternative therapeutic anti-cancer drugs against colon cancer with less toxicity are to be explored . This study was aimed to explore the anti-proliferative and anti-migratory activity of various fractions of Annona reticulata ethanolic leaf extract on human colon cancer cell lines (HCT-116) and to explore the potential molecular targets from the most potent plant extract fraction. Methods After obtaining ethical clearance from the institutional ethics committee, the extract and fractions were prepared and a preliminary analysis of the phytochemical was done qualitatively. Total phenolic and flavonoids were determined. Ethanolic leaf extract and its fractions were subjected to cytotoxicity analysis using the sulforhodamine B assay and the most promising fraction which showed the highest viability was selected to study anti-migratory activity. The anti-migratory effect was studied using a scratch wound healing assay. Gas chromatography-mass spectrometry (GC-MS) was done to identify the major phytocompounds present in the fraction. The major five phytocompounds identified from the GC-MS were subjected to bioinformatics analysis. Result Among the four fractions, the petroleum ether fraction exhibited the highest anti-proliferative activity. The migration of colon cancer cells was significantly inhibited by the extract and petroleum ether fraction. The major phytocompounds identified from GC-MS were phytol (13.03%), 2,6-bis (3,4-methylenedioxyphenyl)-3,7-dioxabicyclo (3.3.0) octane (11.95%), gamma.-sitosterol (10.45%), alpha.-tocopherol-beta.-D-mannoside (7.50%) and 3-amino-4-piperonyl-5-pyrazolone (5.84%). The bioinformatics analysis of these phytochemicals showed a high potential to affect the levels of key proteins driving colon cancer progression, inhibiting the enzymes and proteins overexpressed in cancer. Conclusion The outcome of this study endorses the potential of phytochemicals of the petroleum ether fraction of ethanolic leaf extract of Annona reticulata for the development of a new chemotherapeutic agent in the treatment of colon cancer.

Artemdubinoids A–N: novel sesquiterpenoids with antihepatoma cytotoxicity from Artemisia dubia

In pursuit of effective agents for hepatocellular carcinoma derived from the Artemisia species, this study built upon initial findings that an ethanol (EtOH) extract and ethyl acetate (EtOAc) fraction of the aerial parts of Artemisia dubia Wall. ex Bess. exhibited cytotoxicity against HepG2 cells with inhibitory rates of 57.1% and 84.2% (100 μg·mL−1), respectively. Guided by bioactivity, fourteen previously unidentified sesquiterpenes, artemdubinoids A–N (1–14), were isolated from the EtOAc fraction. Their structural elucidation was achieved through comprehensive spectroscopic analyses and corroborated by the comparison between the experimental and calculated ECD spectra. Single crystal X-ray diffraction provided definitive structure confirmation for artemdubinoids A, D, F, and H. Artemdubinoids A and B (1–2) represented unique sesquiterpenes featuring a 6/5-fused bicyclic carbon scaffold, and their putative biosynthetic pathways were discussed; artemdubinoid C (3) was a novel guaianolide derivative that might be formed by the [4 + 2] Diels–Alder reaction; artemdubinoids D and E (4–5) were rare 1,10-seco-guaianolides; artemdubinoids F–K (6–11) were chlorine-containing guaianolides. Eleven compounds exhibited cytotoxicity against three human hepatoma cell lines (HepG2, Huh7, and SK-Hep-1) with half-maximal inhibitory concentration (IC50) values spanning 7.5−82.5 μmol·L−1. Artemdubinoid M (13) exhibited the most active cytotoxicity with IC50 values of 14.5, 7.5 and 8.9 μmol·L−1 against the HepG2, Huh7, and SK-Hep-1 cell lines, respectively, which were equivalent to the positive control, sorafenib.

Polyphenol-enriched Penthorum chinense Pursh ameliorates alcohol-related liver injury through Ras/Raf/MEK/ERK pathway: Integrating network pharmacology and experiment validation

Ethnopharmacological relevancePenthorum chinense Pursh (PCP) has acknowledged as an edible herbal medicinal plant for the prevention and treatment of alcoholic liver injury (ALI). However, only few of researches focus on the chemical material basis and potential mechanisms of PCP against ALI. Aim of the studyHerein, we explored the therapeutic effects of PCP extract against ALI based on the integration of network pharmacology, molecular docking, and experiment validation. MethodsBased on the standard quality control of PCP herbs by UPLC fingerprint and quantitative determination, 80% ethanol extract fraction of PCP containing more polyphenols, compared to aqueous extract fraction of PCP, were chosen for further experiments. After oral administration of PCP ethanol extract, serum pharmacochemistry based on UPLC-Q-Exactive-MS analysis was implemented to evaluate the potential effective compounds. These absorbed prototypes in PCP were used to construct network pharmacology and predict the potential mechanisms of PCP extract against ALI. Then, the predicted targets and biological mechanisms of PCP extract were validated using animal experiments and molecular docking analysis. ResultsAlthough totally 19 polyphenol compounds were identified in PCP ethanol extract by UPLC-MS analysis, only 18 absorbed prototypes were found in the serum collected from mice at 1 h post-administration with PCP extract. These candidate active compounds were further screened into 13 compounds to construct network pharmacology and 433 targets were identified as PCP targets. GO and KEGG pathway enrichment analyses indicated that the effects of PCP extract would involve in Ras signaling pathway. The animal experiments on chronic ALI model mice shown that the oral administration of PCP can alleviate ALI by attenuating hepatic oxidative stress, inflammation and down-regulating the target proteins in Ras/Raf/MEK/ERK pathway. Molecular docking analysis revealed the good binding ability between the three polyphenols (i.e. quercetin, apigenin, thonningianin B) in PCP with the top contribution in network pharmacology, and these target proteins (Ras, Raf, MEK1/2, and ERK1/2). ConclusionOur results clarified that PCP ethanol extract could effectively alleviate ALI by down-regulating Ras/Raf/MEK/ERK signaling pathway promisingly. Quercetin, apigenin, and thonningianin B may be the active compounds of PCP, attributing to the intervention benefits of PCP against ALI.

Phytochemical profiling and antidiabetic evaluation of Peperomia pellucida as a potential alpha glucosidase inhibitor

Abstract. Hidayati S, Agustin AT, Sari EK, Sari SM, Destiawan RA, Silvana WA. 2023. Phytochemical profiling and antidiabetic evaluation of Peperomia pellucida as a potential alpha glucosidase inhibitor. Biodiversitas 24: 5972-5978. One strategy in post-prandial hyperglycemic control can be done by inhibiting the digestion of dietary carbohydrates through the inhibition of alpha glucosidase enzymes. This study was conducted to determine the antidiabetic activity of Peperomia pellucida in inhibiting the alpha glucosidase. An inhibitory activity analysis test was carried out on alpha glucosidase in vitro and in silico using alpha glucosidase. Molecular docking between receptors and ligands was performed using Hex 8.0 software. The setting column is set in Shape+Electro+DARS mode. The results showed ethanol extract and ethyl acetate fraction in vitro showed the ability to inhibit the activity of alpha glucosidase enzyme with C50 values of 13.43 mg/mL and 9.73 mg/mL respectively with IC50 positive control acarbose values of 8.11 mg/mL. The results of in silico analysis showed that the Patuloside A component was able to bind to the binding site of alpha glucosidase and gave the smallest binding energy value with a value of -321.4 kcal/mol compared to isovitexin, isoswertisin, pellucidatin, and caryatin-7-O-?-rhamnoside compounds. P. pellucida has the potential to be developed as an antidiabetic agent that has activity in inhibiting the work of the enzyme alpha glucosidase so that it can reduce blood glucose levels.

Perspectives on antimicrobial properties of Paulownia tomentosa Steud. fruit products in the control of Staphylococcus aureus infections

Ethnopharmacological relevancePaulownia tomentosa Steud. (P. tomentosa) is a medium-sized tree traditionally used in Chinese folk medicine for the treatment of infectious diseases. It is a rich source of prenylated phenolic compounds that have been extensively studied for their promising biological activities. Aim of the studyDue to the increasing development of antibiotic resistance, our study investigated plant-derived natural products from the fruits of P. tomentosa that could control Staphylococcus aureus infections with novel targets/modes of action and reduce antimicrobial resistance. Materials and methodsThe ethanolic extract was fractionated and detected by liquid chromatography. The antistaphylococcal effects of the plant formulations were studied in detail in vitro by various biological methods, including microdilution methods for minimum inhibitory concentration (MIC), the checkerboard titration technique for synergy assay, fluorescence measurements for membrane disruption experiments, autoinducer-2-mediated bioassay for quorum sensing inhibition, and counting of colony-forming units for relative adhesion. Morphology was examined by transmission electron microscopy. ResultsTotal ethanolic extract and chloroform fraction showed MICs of 128 and 32 μg/mL, respectively. Diplacol, diplacone, and 3′-O-methyl-5′-hydroxydiplacone inhibited S. aureus growth in the range of 8–16 μg/mL. Synergistic potential was shown in combination with mupirocin and fusidic acid. The ethanolic extract and the chloroform fraction destroyed the cell membranes by 91.61% and 79.46%, respectively, while the pure compounds were less active. The ethanolic extract and the pure compounds reduced the number of adhered cells to 47.33–10.26% compared to the untreated control. All tested plant formulations, except diplacone, inhibited quorum sensing of S. aureus. Transmission electron microscopy showed deformation of S. aureus cells. ConclusionsThe products from the fruit of P. tomentosa showed antimicrobial properties against S. aureus alone and in combination with antibiotics. By affecting intracellular targets, geranylated flavonoids proposed novel approaches in the control of staphylococcal infections.

Cytotoxic Test of Fig Leaf Ethanol Extract Fraction (Ficus carica L.) Using Brine Shrimp Lethality Test Method

Cytotoxic test of fig leaf ethanol extract fraction (Ficus carica L.) has been conducted with the Brine Shrimp Lethality Test (BSLT) method. This study aims to determine the cytotoxic activity of fig leaf ethanol extract using the Brine Shrimp Lethality Test (BSLT) method. Fig leaves are extracted by maceration using 70% ethanol, then proceed with fractionation based on polarity, fractionation using N-hexane solvent, ethyl acetate and water. The results of phytochemical screening of fig leaf ethanol extract fraction stated that, n-hexan fraction contains flavonoid, phenolic, triterpenoid and saponin compounds. The ethyl acetate fraction contains phenolic compounds, triterpenoids and saponins. While the water fraction contains flavonoids, phenolics and saponin compounds. The results of cytotoxic testing of the fraction of fig leaf ethanol extract obtained LC50 values in the n-hexane fraction were 41.78 ppm (toxic), ethyl acetate fraction 33.57 ppm (toxic), and water fraction 16.36 ppm (very toxic).

Characterization of phenolic compounds in Eugenia uniflora leaves by ESI(-) FT-ICR MS, analysis of cytotoxic activity on gastric adenocarcinoma (AGS cells), and anti-Helicobacter pylori activity

Eugenia uniflora leaves are a source of flavonoids and ellagitannins, and the Brazilian population uses them to treat various diseases, including gastrointestinal disorders. This study aimed to determine if the ethanol extract and other derivatives are effective cytotoxic agents against gastric adenocarcinoma (AGS cells) and anti-H. pylori agent, its chemical composition, and mass spectrometry characterization of the more abundant compounds. The results were compared with the literature. The aqueous fraction, rich in Oenothein B and Gemin D/Hippomanin A, showed anti-H. pylori activity and higher cytotoxicity on AGS cells compared to the other samples analysed. Furthermore, the ESI(-) FT-ICR MS characterized the more abundant phenolic compounds, including Quinic Acid, Myricitrin, Gemin D/Hippomanin A, and Oenothein. Therefore, the activity of the ethanolic extract and aqueous fraction for gastric cancer and against H-pylori seems to originate from the antiproliferative and bacteriostatic effects of tannins and flavonoids.

Evaluation of Insecticidal Performances of Jayanti Plant (Sesbania sesban) for Integrated Control of Cabbage Caterpillar (Plutella xylostella)

The study on the use of local natural materials as a source of insecticides is intended, among other things, to reduce farmers' dependency on using synthetic chemicals. This research was conducted to obtain the most selective insecticidal active ingredient from Sesbania sesban plant for controlling cabbage caterpillars, choose a target mode of action that has a safe impact on the environment, and determine insecticidal stability during storage of the material prior to application and its stability in water after application. The dried powder of S. sesban leaves was extracted in stages using a series of solvents, namely hexane, DCM, ethanol and water, respectively. Each of the extract fractions produced was tested for its lethal toxicity against Plutella xylostella larvae and Diadegma semiclausum imago. Each of mortality data were processed by probit analysis to produce LC50(s) to determine their insecticidal selectivity. Subsequent bioassays were carried out using extract-water of S. sesban leaves and the data were processed by probit analysis to determine their anti-ovipositor, ovicidal powers and anti-feedant properties against P. xylostella. The insecticidal stability of S. sesban was also studied according to variations in the storage time of the simplicia before extraction, the storage time of the extract before dissolution, and its stability in water during application. The data was processed using ANOVA. The results show that the insecticidal activity of the ethanol extract fraction of S. sesban leaves for controlling cabbage caterpillars was very selective, namely very toxic to P. xylostella but very less toxic to D. semiclausum. Against P. xylostella, S. sesban leaf extract has also been proven to significantly prevent oviposition, inhibit egg hatching and feeding activity. The insecticidal power of S. sesban did not decrease significantly during storage of the simplicia for less than three months and during storage of the extract for less than seven days. However, the toxicity of the extract solution decreased drastically after 24 hours of application

Bioactive compounds from the leaves of Maytenus ilicifolia Mart. ex Reissek: Inhibition of LDL oxidation, glycation, lipid peroxidation, target enzymes, and microbial growth

Ethnopharmacological relevanceMaytenus ilicifolia Mart. ex Reissek, a medicinal plant used for treating gastritis, ulcers, and gastric disorders, possesses therapeutic properties attributed to diverse leaf compounds—terpenoids, alkaloids, flavonoids, phenols, and tannins, reflecting the ethnopharmacological knowledge of traditional users. Aims of the studyWe aimed to assess the antioxidant and antiglycant capacities of Maytenus ilicifolia's ethanolic extract and organic fractions, identify bioactive compounds through HPLC-MS/MS analysis, and conduct phytochemical assessments. We also assessed their potential to inhibit digestive and cholinesterase enzymes, mitigate oxidation of human LDL and rat hepatic tissue, and examine their antimicrobial and cytotoxic properties. Materials and methodsOrganic fractions (hexane - HF-Mi, dichloromethane - DMF-Mi, ethyl acetate - EAF-Mi, n-butanol - BF-Mi, and hydromethanolic - HMF-Mi) were obtained via liquid-liquid partitioning. Antioxidant (DPPH, FRAP, ORAC) and antiglycant (BSA/FRU, BSA/MGO, ARG/MGO/LDL/MGO models) capacities were tested. Phytochemical analysis employed HPLC-MS/MS. We also studied the inhibitory effects on α-amylase, acetylcholinesterase, butyrylcholinesterase, human LDL and rat hepatic tissue oxidation, antimicrobial activity, and cytotoxicity against RAW 264.7 macrophages. ResultsHPLC-ESI-MS/MS identified antioxidant compounds such as catechin, quercetin, and kaempferol derivatives. Ethanolic extract (EE-Mi) and organic fractions demonstrated robust antioxidant and antiglycant activity. EAF-Mi and BF-Mi inhibited α-amylase (2.42 μg/mL and 7.95 μg/mL) compared to acarbose (0.144 μg/mL). Most organic fractions exhibited ∼50% inhibition of acetylcholinesterase and butyrylcholinesterase, rivaling galantamine and rivastigmine. EAF-Mi, BF-Mi, and EE-Mi excelled in inhibiting lipid peroxidation. All fractions, except HMF-Mi, effectively countered LDL oxidation, evidenced by the area under the curve. These fractions protected LDL against lipid peroxidation. ConclusionThis study unveils Maytenus ilicifolia's ethanolic extract and organic fractions properties. Through rigorous analysis, we identify bioactive compounds and highlight their antioxidant, antiglycant, enzyme inhibition, and protective properties against oxidative damage. These findings underline its significance in modern pharmacology and its potential applications in healthcare.

Determination of Flavonoid Content and Anti-Inflammatory Activity Extract and Fraction of Sungkai Leaf (Peronema canescens Jack)

The SARS-CoV-2 virus is the infectious agent that causes COVID-19, a feverish condition brought on by inflammation in the infected patient’s body. Sungkai leaf (Peronema canescens Jack) is one of the Indonesian people who rely on herbal remedies to treat COVID-19. This study aims to determine how much flavonoids are present in ethanol, ethyl acetate, and n-hexane extracts, as well as the anti-inflammatory properties of ethanol extract, ethanol fraction, and n-hexane fraction of Sungkai leaves. The procedure was performed in-vitro with a UV-Visible spectrophotometer by observing the absorption response to inhibition of denaturation of inflammatory protein. The inhibition value was then computed via linear regression, and the IC50 and IC70 values were ascertained afterward. The ethanol fraction, ethanol extract, and n-hexane fraction in this investigation had the best inhibition values (%) at a concentration of 15 ppm, corresponding to 74.27%, 54.48%, and 18.52%. The sungkai leaves ethanol fraction > n-hexane fraction > ethanol extract had the best IC50 and IC70 values. Comparatively, the ethanol extract contained the largest amounts of flavonoids, 38.782 μg/mL.

Sapindus saponaria (Sapindaceae): Chemical Composition and Toxic Effect on Artemia salina (Artemiidae) and Aedes aegypti (Culicidae)

Purpose: Phytochemical analysis of Sapindus saponaria leaves extracts, evaluating their effect on Artemia salina and Aedes aegypti larvae. Theoretical framework: Dengue is a disease transmitted by Aedes aegypti mosquitoes and considered a serious public health problem. Its control is done through the management of adult forms of A. aegypti, which occurs through the application of synthetic insecticides. However, its constant use has selected populations of insects resistant to chemical products and for this reason alternative solutions are sought, such as the use of phytoinsecticides. Method/design/approach: Leaves were collected in areas of native vegetation, from which the ethanolic, aqueous and acetonitrile extracts were obtained, in addition to the ethanolic extract fractions, submitted to phytochemical analysis, with A. aegypti and A. salina larvae used to evaluate the toxic effect of extracts and fractions. Results and conclusion: The ethanol extract showed the greatest diversity of secondary metabolites, the best CL90 and 100% larval mortality, negatively affecting larval time and larval and pupal mortality, in addition to decreasing the number of formed alates. The inhibitory effect is probably linked to the presence of cardiotonic saponins and heterosides, known for their deleterious action in organisms. Research implications: The research indicates the potential use of Sapindus saponaria leaves in the control of the dengue vector. Originality/value: Viability of plant strata for vector control, reducing the application of synthetic insecticides and improving environmental quality.

Đánh giá tác dụng chống oxy hóa và bảo vệ gan trên thực nghiệm của cây muồng lùn

Antioxidant and hepatoprotective effects of different fractions of extract from Chamaecrista pumila (Lam.) K. Larsen were investigated by employing in vitro and in vivo models, respectively. In vitro antioxidant activity of ethanol extract and its n-hexan, ethyl acetate, chloroform, n-butanol and remained water fraction extracts obtained from C. pumila was assessed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide radical scavenging assay. In vitro results revealed that the ethanol extract had potential DPPH radical and superoxide radical scavenging activities with an IC50 value of 18.65 (9.91-35.08) µg/ml and 12.33 (5.24-29.02) µg/mL, respectively. Among fractions extracted from ethanol extract, the ethyl acetate fraction possessed highest antioxidant property through DPPH radical scarvenging activity with an IC50value of 9.98 (6.23-16.01) µg/mL and superoxide radical scavenging activity with an IC50 value of 8.38 (6.11-11.50) µg/mL. In vivo hepatoprotective activities of ethanolic extract and ethyl acetate fraction obtained from C. pumila were investigated in mice using a model of paracetamol-induced hepatotoxicity. Ethanol extract of C. pumila at doses of 250 mg/kg and 500 mg/kg decreased levels of serum transaminases ASAT. Ethyl acetate fraction of C. pumila at doses of 250 mg/kg and 500 mg/kg significantly improved the parameters of paracetamol-induced liver injury including ASAT, ALAT and superoxide dismutase (SOD) activities; ethyl acetat fraction of C. pumila at dose of 500 mg/kg also exhibited in vivo antioxidant properties by reducing malondialdehyde (MDA) content and increasing glutathione (GSH) content.

Evaluation of the acute toxicity and antidiabetic activity of Coutoubea spicata “nicolau” ethanolic extract and ethyl acetate fraction

In order to understand antidiabetic potential and toxicity, this study aimed to evaluate the acute toxicity and antidiabetic activity of ethanolic extract and ethyl acetate fraction obtained from Coutoubea spicata “nicolau” shoots. Chemical constituents and acute toxicity were investigated. In alloxan-induced diabetic rats, extract and fraction were tested at dose of 100 mg/kg, p.o. Body weight gain, glucose, lipid profile and oxidative stress markers in serum and tissues were determined. In vitroantioxidant activity was performed. Swertiamarin, gentiopicrin, deoxyloganic acid, clovin and robinin, and their p-coumaric ester were identified. Extract and fraction were classified as safe (category 5). In diabetic rats, Coutoubea spicata reduced glycaemia, which was accompanied by body weight recovery gain and attenuation in oxidative stress markers. Fraction showed scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical (ABTS) radicals and reducing power higher than that of the extract. Extract and fraction of Coutoubea spicata didn’t present significant toxicity and it can be investigated as a therapeutic alternative in diabetes.

Uji Aktivitas Antioksidan Ekstrak dan Fraksi n-Heksan, Etil asetat, dan Air Kulit Jambu Biji Australia (Psidium guajava L.) Dengan Metode DPPH (1,1-Difenil-2-Pikrilhidrazil)

Free radicals work by binding to molecules or cells in the body and are very dangerous because they can damage cells in the body and trigger disease. Antioxidants are compounds that can counteract the effects of free radicals. This study aims to determine the antioxidant activity of ethanol extract and the n-hexane, ethyl-acetate and water fractions of Australian guava peel (Psidium guajava L.). Extraction was carried out by maceration method using 70% ethanol. The skin of the Australian guava (Psidium guajava L.) contains alkaloids, flavonoids, tannins, triterpenoids and saponins. Fractionation was carried out with solvents based on polarity differences obtained by the n-hexane fraction, ethyl acetate fraction, and air fraction. Antioxidant test by inhibiting DPPH free radicals (1,1-Diphenyl-2-Picrylhydrazyl) on ethanol extract and n-hexane fraction, ethyl acetate fraction and air at several concentrations, namely 50 ppm, 100 ppm, 150 ppm, 200 ppm, and 250 ppm. The IC50 calculation results obtained for the ethanol extract were 4.406 µg/mL, the n-hexane fraction was 4.131 µg/mL, the ethyl acetate fraction was 3.812 µg/mL and the air fraction was 3.312 µg/mL. The antioxidant activity with the best IC50 value was the fraction air.

Anthelmintic efficacy of Holarrhena pubescens against Raillietina spp. of domestic fowl through ultrastructural, histochemical, biochemical and GLCM analysis.

Globally, traditional knowledge systems are a powerhouse of information which can revolutionise the world, if decoded accurately and logically. Plant-based ethno-traditional and folklore curatives/medicines has a firm basis in the psyche of the common masses of West Bengal and Holarrhena pubescens is a representative example of it. This article communication on depicting the anthelmintic efficacy of ethanolic extract and Ethyl acetate fraction of the stem bark of Holarrhena pubescens against the cestode Raillietina spp. through efficacy studies, ultra-structural observations, histochemical and biochemical analysis on some tegumental enzymes i.e., Acid Phosphatase (AcPase), Alkaline Phosphatase (AlkPase), Adenosine Triphosphatase (ATPase) and 5'-Nucleotidase (5'-Nu) along with Gray Level Co-occurrence Matrix (GLCM) analysis of histochemical study. Praziquantel was used as the reference drug. Investigations revealed 10mg/ml dosage of crude extract was the most efficacious dose and amongst the fractions the ethyl acetate fraction showed the most anthelmintic property. Ultrastructural studies through Scanning Electron Microscope (SEM) and Transmission Electron Microscope (TEM) clearly depicted the damage in head, sucker, proglottids, proximal and distal cytoplasm (DC), microtriches (MT), basal lamina (BL), nuclear membrane (NM), and, nucleolus (NL) in the treated worms. Histochemical studies revealed decrease in staining intensity for all the tegumental enzymes in the treated worms compared to control. The GLCM analysis strongly supported the result of histochemical studies. Biochemical studies revealed marked reduction in enzyme activity in the treated worms with maximum reduction in the activity of 5'- Nu (77.8%) followed by ATPase (63.17%).

Evaluation of the Anti-Diabetic Activity of Paliasa Leaf Ethanol Fraction (Kleinhovia hospita L) Through Inhibition of the Enzymes $\alpha$-Glucosidase and $\alpha$-Amylase

Kleinhovia hospita has long been used to treat diseases, including diabetes mellitus. This study aimed to evaluate if the hypoglycemic effects of K. hospita extract and fractions were related to its inhibition on the activity of $\alpha$-amylase and $\alpha$-glycosidase enzymes. K. hospita leaves were extracted with 96% ethanol and then subjected to liquid-liquid partition using n-hexane, ethyl acetate, and water as solvents. The inhibitory capacity of ethanolic extract and its fractions were tested against $\alpha$-amylase and $\alpha$-glycosidase enzyme activity and compared to acarbose. The in vitro study revealed that the IC50 of ethanolic extract, hexane, ethyl acetate, and water fractions were 1496,8 µg/mL, 1266,7 µg/mL, 12577,5 µg/mL, and 5217,0 µg/mL, respectively, against alfa-amylase activity. Whereas, for $\alpha$-glycosidase activity, the IC50 values were 1341,2 µg/mL, 25470,0 µg/mL, 64450,0 µg/mL, and 1291,4 µg/mL, respectively. These values were markedly greater than those of the acarbose (7,4 and 5,5 µg/mL, respectively). From the research results, it was concluded that K. hospita leaf extract had a high IC50 value from K. hospita extracts and fractions, which indicated that the hypoglycemic effect of K. hospita was not related to the inhibition of the $\alpha$-amylase and $\alpha$-glycosidase enzymes.