Fractionation Of Ethanolic Extract Research Articles

In vitro Antibacterial and Cytotoxic Activities of Various Fractions of Lippia alba (Mill.)

Lippia alba (Mill.) (Family: Verbenaceae) is widely known due to both ethnobotanical uses and chemical diversity. The present work was designated to determine the antimicrobial and cytotoxic activities of the aqueous ethanolic extract and various fractions of this plant. The whole plant was made into coarse powder and was extracted with 5% aqueous ethanol. The concentrated ethanolic extract (abbreviated as LAE) was successively partitioned with n-hexane (LAH), chloroform (LAC), ethyl acetate (LAA), and the remaining part was designated as water (LAQ). The antimicrobial and cytotoxic activities of extract and all these fractions were evaluated in vitro using disc diffusion technique and brine shrimp (Artemia salina) lethality assay, respectively. In antimicrobial assay, five Gram-positive and five Gram-negative bacteria and standard antibiotic ciprofloxacin (10 μg/disc) were used. All the samples were tested at four concentrations (200, 400, 600 and 800 μg/disc). Our study showed that the extractive and different fractions of L. alba had antibacterial activity against all the tested bacteria except Pseudomonas aeruginosa. Among the extractives, LAE and LAH had significant antibacterial activity when compared to standard ciprofloxacin. The zone of inhibition of LAE and LAH were found to be 14.4, 24.4, 13.4, 15.2, 18.5, 24.1, 16.1, 23.5, 23.2 and 14.5, 20.1, 12.5, 17.1, 17.5, 20.1, 12.5, 19.4, and 18.16 mm against Bacillus cereus, Staphylococcus aureus, Streptococcus-β- haemolyticus, Bacillus subtilis, Sarcina lutea, Escherichia coli, Klebsiella pneumonia, Salmonella typhi and Shigella dysenteriae, respectively, at 800 μg/disc. The median lethal concentration (LC50) in brine shrimp lethality bioassay was determined by extrapolation from graph and the values were found to be 41.9, 23.9, 21.3, 88.7, 6.7 and 7.5 μg/ml for LAQ, LAH, LAC, LAA, LAE and standard vincristine sulfate, respectively. In the study, different mortality rate was observed at different concentration and all the samples showed positive response, indicating the presence of cytotoxic components in L. alba. Phytochemical screening revealed that L. alba contains saponins, tannins, glycosides, steroids, alkaloids, terpenoids and flavonoids. The results of the study indicate that L. alba might be a good source of potent antibiotic and anticancer drugs. Bangladesh Pharmaceutical Journal 26(2): 150-156, 2023 (July)

Antiplatelet and thrombolytic activity of phenolic-insistent fractions from the new-fangled stem of Ficus benghalensis with concurrent GC-MS analysis

Ficus benghalensis is the ingredient of a variety of Ayurvedic herbal formulations for the management of blood-related illnesses. In the current study, the new-fangled stem ethanol extract fractions in chloroform and methanol (CFFB & MFFB) were assessed for antiplatelet, thrombolytic and toxicity studies, as well as for phytoconstituent identification GC/MS was performed. The dried powdered stem bud was extracted with 80% ethanol and successively fractionated by chloroform and methanol (CFFB & MFFB). The anti-platelet, anti-thrombotic, and thrombolytic, activity of CFFB & MFFB were tested in ex vivo mode and toxicity of methanol fraction (MFFB) was tested in in vivo. The chief feasible marker components for antiplatelet activity recognized by GC-MS in the MFFB are Diethyl phthalate, (E)-4-(3-Hydroxyprop-1-en-1-yl)-2-methoxyphenol, 7,9-Di-tert-butyl-1-oxaspiro (4,5) deca-6,9-diene-2,8-dione and trans-Sinapyl alcohol might act as irreversible cyclooxygenase inhibitors like Aspirin. In the study, at 50 ​μg/mL, the antiplatelet activity of CFFB, MFFB, and aspirin was 50.41, 82.19, and 86.34%, and a substantial adjournment in clot development was observed whereas CFFB at different dosages did not exhibit significant outcome on the adjournment of clot formation, antiplatelet, and antioxidant activity. The toxicity examination of MFFB did not confirm any substantial signs of toxicity and mortality up to 1.5 ​g/kg, b.w and non-toxic up to 1.0 ​g/kg, b.w which is capable of the comportment of atherothrombotic ailments. The MFFB exhibited anti-platelet, anti-thrombotic, thrombolytic, and anti-oxidant activity, and capacity to prevent cardiovascular disorders without causing toxicity.

Phytochemical screening: Determination of total polyphenol and flavonoid contents, antioxidant and antimicrobial activity of leaves and seeds of Abelmoschus esculentus L.

Reputed to be a plant rich in antioxidants, Abelmoschus esculentus or okra is a very effective vegetable against diabetes. In this work, we made a qualitative study of the bioactive compounds contained in the leaves and seeds of okra, through a phytochemical screening. Subsequently, the antioxidant and antimicrobial activities of these extracts were studied. The Folin-Ciocalteu reagent was used to evaluate the content of the phenolic compound. The DPPH● radical made it possible to measure the antioxidant power of the extracts. The phytochemical analysis made it possible to high the presence of substances with great therapeutic values ​​(polyphenols, flavonoids, tannins, alkaloids, etc.). The antibacterial activity was measured on bacterial strains. The antioxidant capacities reveal that the extracts of the leaves and seeds of okra possess an interesting anti-free radical activity with IC50 values ​​of 06 ± 0.21 mg / mL and 8.85 ± 0.28 mg / mL respectively for the leaves and the seeds. . Biological analysis shows that the E. faecalis strain is resistant to different extracts. E. coli, S. aureus, and P. aeruginosa strains show some sensitivity to ethanolic extracts. However, E. colis and S. aureus strains show the best antimicrobial activities with a MIC of 3.75 mg/mL for crude ethanol extract and ethanol fraction of leaves and seeds.

Modulation of IL-6 in ameliorating arthritic conditions in rats by Rosa alba L flower extract and fraction

Introduction: rheumatoid arthritis characterized by joint inflammation and pain,affecting millions of peoples around the world. Traditional system of medicine hadproven to be preventive and treating physical and mental illness. Objective: theobjective of the study is to assess anti-arthritic potential of the plant Rosa alba L byconsidering CFA, formaldehyde and turpentine oil induced arthritic model. Materialand Method: ethanolic extract and its ethyl acetate fraction were considered for thestudy and quercetin was quantitatively estimated by high performance thin layer chromatographic(HPTLC) method. Moreover, hematological and biochemical studies(IL-6) were performed in blood and serum of wistar albino rat. Then histopathologicalstudies had also been performed on rat hind paw joint. Result: phytochemicalscreening estimated the presence of carbohydrates, phenolic compounds, flavonoids,phytosterols, amino acids and fixed oil in ethanolic extract and ethyl acetate fraction ofthe plant. Amount of quercetin in the flowers of the plant was found to be 0.26% w/wby quantitative HPTLC method. The level of proinflammatory cytokines, interleukin(IL-6) was considerably decreased (p<0.001) in the fraction treated group of rats at400 mg/kg b.w by considering complete freund’s adjuvant (CFA) induced arthritic model. Conclusion: Ameliorated hematological,hematological, biochemical and histopathologicalparameters confirmed the antiarthritic potential of the plant.

Studies on the Nonalkaloidal Secondary Metabolites of Hippeastrum vittatum (L'Her.) Herb. Bulbs.

Sixteen chemically varied metabolites were isolated from the bulbs of Hippeastrum vittatum (L'Her.) Herb., including eight flavonoids [3'-methyl isoliquiritigenin (2), 7-hydroxyflavan (8), 7-hydroxyflavanone (9), 7-hydroxyflavan-3-ol (10), 7-methoxy-3',4'-methylenedioxyflavan-3-ol (11), 7-hydroxy-3',4'-methylenedioxy flavan (12), 2',4'-dihydroxy-3'-methyl-3,4-methylenedioxychalcone (13), and isoliquiritigenin (14)], four acetophenones [2,6-dimethoxy-4-hydroxyacetophenone (3), 2,4-dihydroxyacetophenone (4), 2,4-dihydroxy-6-methoxy-3-methylacetophenone (6), and 2,4,6-trimethoxyacetophenone (7)], two alkaloids [lycorine (1) and narciprimine (15)], one phenol derivative [p-nitrophenol (5)], and one steroid [β-sitosterol 3-O-β-glucopyranoside (16)]. Their structures were elucidated by combining one- and two-dimensional NMR and ESI-MS techniques and by comparison with the reported literature data and some authentic samples. Except for lycorine (1), the isolated metabolites were obtained herein for the first time from Hippeastrum plants, among which compound 13 was identified as a new chalcone derivative. Additionally, the total phenolic and flavonoid contents of the total ethanol extract and different fractions of the bulbs were determined by the Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively, whereas their antioxidant potential was compared using the phosphomolybdenum and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assays. Finally, the binding affinities of compounds 1-16 to some key target proteins of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), namely, main protease (Mpro), papain-like protease (PLpro), and RNA-dependent RNA polymerase (RdRp), were screened and compared using molecular docking analysis. The possible chemotaxonomic significance of the identified metabolites was also discussed.

<i>Invitro</i> Antitrypanosomal activity of fractions of ethanol extract from <i>Terminalia cattapa</i> leaf against <i>Trypanosomal Brucei Brucei</i>

Trypanosomiasis is a tropical disease threatening human and animal health. Trypanosoma brucei brucei is unicellular parasites transmitted by the bite of tsetse fly and is the causative agent of sleeping sickness in humans and related diseases in animals. Over 25% of our common medicines contains some compounds obtained from plants. This research work is aimed at evaluating the invitro activity of fractionated ethanol extract of Terminalia cattapa leaf against Trypanosoma brucei brucei. Crude extract was obtained by cold maceration using absolute ethanol, followed by partial purification of the extract using column chromatography techniques. The test was performed in triplicates using a 96 well microtitre plate. Light microscope was used to view the parasite motility at magnification of × 400. Treatment with varying concentrations of pooled fractions F1, F2 and F3 (100, 50 and 25 mg/ml) resulted in reduction of number of motile parasites compared to the negative control loaded with parasites. In conclusion, fractions (F2 ans F3) from ethanol extracts of Terminalia cattapa leaf have anti-trypanosoma activities against Trypanosoma brucei brucei in a dose depended manner with F3 being the most potent..

Bioassay guided fractionation of anthelmintic bioactive compounds from surinam cherry (Eugenia uniflora L.) fruits

Bio-fractionation process is the most reliable protocol, which enables the screening and purification of phytoconstituents from the crude extracts efficiently. Soil transmitted helminths induce the most challenging health concerns, which need to be addressed in a holistic way for natural product utilization. Surinam cherry fruits, rich in phytonutrients and secondary metabolites with several health benefits are also reported to have anthelmintic potential. Bioassay guided fractionation of Surinam cherry mature green pulp ethanol (MPE) extract afforded five active fractions with higher anthelmintic activity against all the stages of Caenorhabditis elegans and Heterorhabditis indica than their crude extract. The active fractions were further characterized using UPLC-HRMS/MS analysis and FTIR. Fraction C3 obtained using 100% chloroform resulted in 97.67% inhibition of egg hatching, 95% larval death with APT of 8.08 min, and 95.33% adult worm death with APT of 4 min at the concentration of 2 mg/ml in C. elegans. The anthelmintic activity of C3 was correlated to 14 phenolic compounds such as kaempferol-3-O-glucuronide, kaempferol-3-O-sophoroside, isorhamnetin, rhamnetin, isorhamnetin-3-O-glucoside, catechin, eriodictyol-7-O-glucoside, luteolin-6-C-glucoside, 4-hydroxyphenylacetic acid, estragole, biochanin A, syringaresinol, juglone, and pterostilbene present in it. This study indicates that relatively more active fractions could be separated from Surinam cherry MPE crude extract for anthelmintic potential against C. elegans and H. indica, which can be used for pharmaceutical applications.

Antibacterial Activity of Ethanol Extract, n-Hexane and Ethyl Acetate Fraction of Mundar (Garcinia forbesii) Pericarp

Garcinia forbesii is traditionally used as medicine and cosmetics. The aimed was to determine antibacterial activity of ethanolic extract and fraction of Garcinia forbesii pericarp against Staphylococcus aureus and Propionibacterium acne based on the diameter of inhibition zone, the minimum inhibitory concentration (MIC) and the minimum killing concentration (MBC). The results showed that the largest inhibition zone diameter against Staphylococcus aureus and Propionibacterium acne was indicated by ethyl acetate fraction of 5.08±1.020 mm and 14.33±3.326 mm. The MIC value of ethanol extract against Staphylococcus aureus and Propionibacterium acne bacteria was >1.5%, n-hexane and ethyl acetate fraction had the same results, namely against Staphylococcus aureus >1.5% and Propionibacterium acne at 1.5%. The MBC value was only obtained for fraction samples of Propionibacterium acne at concentration of 1.5%. It was concluded that ethyl acetate had strong antibacterial activity against Propionibacterium acne based on the value of inhibition zone diameter, MIC, and MBC.

"Methanol Fraction of Ethanol Extract of Dialium Guineense Stem Bark Mitigates STZ-Induced Oxidative Stress in Rat Liver"

"Methanol Fraction of Ethanol Extract of Dialium Guineense Stem Bark Mitigates STZ-Induced Oxidative Stress in Rat Liver"

Ethyl gallate isolated from phenol-enriched fraction of Caesalpinia mimosoides Lam. Promotes cutaneous wound healing: a scientific validation through bioassay-guided fractionation.

The tender shoots of Caesalpinia mimosoides Lam. are used ethnomedically by the traditional healers of Uttara Kannada district, Karnataka (India) for the treatment of wounds. The current study was aimed at exploring phenol-enriched fraction (PEF) of crude ethanol extract of tender shoots to isolate and characterize the most active bio-constituent through bioassay-guided fractionation procedure. The successive fractionation and sub-fractionation of PEF, followed by in vitro scratch wound, antimicrobial, and antioxidant activities, yielded a highly active natural antioxidant compound ethyl gallate (EG). In vitro wound healing potentiality of EG was evidenced by a significantly higher percentage of cell migration in L929 fibroblast cells (97.98 ± 0.46% at 3.81μg/ml concentration) compared to a positive control group (98.44 ± 0.36%) at the 48th hour of incubation. A significantly higher rate of wound contraction (98.72 ± 0.41%), an elevated tensile strength of the incised wound (1,154.60 ± 1.42g/mm2), and increased quantity of connective tissue elements were observed in the granulation tissues of the 1% EG ointment treated animal group on the 15th post-wounding day. The accelerated wound healing activity of 1% EG was also exhibited by histopathological examinations through Hematoxylin and Eosin, Masson's trichome, and Toluidine blue-stained sections. Significant up-regulation of enzymatic and non-enzymatic antioxidant contents (reduced glutathione, superoxide dismutase, and catalase) and down-regulation of oxidative stress marker (lipid peroxidation) clearly indicates the effective granular antioxidant activity of 1% EG in preventing oxidative damage to the skin tissues. Further, in vitro antimicrobial and antioxidant activities of EG supports the positive correlation with its enhanced wound-healing activity. Moreover, molecular docking and dynamics for 100ns revealed the stable binding of EG with cyclooxygenase-2 (-6.2kcal/mol) and matrix metalloproteinase-9 (-4.6kcal/mol) and unstable binding with tumor necrosis factor-α (-7.2kcal/mol), suggesting the potential applicability of EG in inflammation and wound treatment.

ANTIHYPERGLYCHEMIC ACTIVITY TEST OF EXTRACT, METHANOL FRACTION AND N-HEKSAN FRACTION OF KETAPANG LEAF (Terminalia catappa) IN ALLOXAN-INDUCED WHITE RATS

Background: Ketapang leaf (Terminalia catappa) can be used as a traditional medicine that has the potential activity to reduce blood glucose levels in the body. Objective: To determine the activity of extracts and fractions of ketapang leaves in reducing blood glucose levels of rats induced by alloxan. Methods: This study was an experimental study using 30 male wistar rats as test animals divided into 6 groups. Group I was healthy control, group II was negative control (K-) that given 0.5% Na CMC solution, group III was positive control (k+) that given metformin suspension, groups IV, V, and VI were given ketapang leaf extract, methanol fraction and n-hexane fraction, respectively. Results: The ethanol extract and methanol fraction of ketapang leaves can reduce blood glucose levels in white rats because they contain flavonoid compounds which are thought to have antihyperglycemic activity. Conclusions: The methanol fraction of ketapang leaves (Terminalia catappa) was most effective in reducing blood glucose levels in alloxan-induced white rats.

Evaluation of in vitro antimicrobial activity of Camellia sinensis leaves collected at different time of harvest

The amounts of chemicals in green tea depend on the time of harvest. In Korea, there is a tendency to harvest tea leaves to make green tea from around April 20 to the end of May. Therefore, we investigated the influence of harvest time of Camellia sinensis leaves on antimicrobial activity. The ethyl acetate and water fraction of ethanol extract from C. sinensis leaves collected every two weeks (April 30, May 14 and May 28, 2021) were tested against 4 Gram-positive and 3 Gram-negative bacteria and one yeast. Antimicrobial activity was screened for clear zone and minimal inhibition concentration (MIC) values determined by disc diffusion method. The ethyl acetate fraction was potentially effective against Methillin-resistant Staphylococcus aureus (MRSA), Bacillus cereus, Staphylococcus aureus and Salmonella typhimurium at all the collection dates. The results showed the diameter of clear zone was proportional to the concentration of tested fractions. The antimicrobial activity of tea leaves was different by collection dates, the quantitative difference was not big. The MIC value of ethyl acetate fraction of tea leaves for Bacillus cereus and B. subtilis was 0.5 mg/ml, on the other hand, the MIC value of the water fraction of tea leaves for B. subtilis, MRSA, Pseudomonas fluorescens and Salmonella typhimurium was 30.0 mg/ml, regardless of collection date. The antimicrobial properties of C. sinensis leaves encouraged an innovative potential application in agricultural, pharmaceutical and food fields.

Evaluation of in vitro antimicrobial activity of Camellia sinensis leaves collected at different time of harvest

The amounts of chemicals in green tea depend on the time of harvest. In Korea, there is a tendency to harvest tea leaves to make green tea from around April 20 to the end of May. Therefore, we investigated the influence of harvest time of Camellia sinensis leaves on antimicrobial activity. The ethyl acetate and water fraction of ethanol extract from C. sinensis leaves collected every two weeks (April 30, May 14 and May 28, 2021) were tested against 4 Gram-positive and 3 Gram-negative bacteria and one yeast. Antimicrobial activity was screened for clear zone and minimal inhibition concentration (MIC) values determined by disc diffusion method. The ethyl acetate fraction was potentially effective against Methillin-resistant Staphylococcus aureus (MRSA), Bacillus cereus, Staphylococcus aureus and Salmonella typhimurium at all the collection dates. The results showed the diameter of clear zone was proportional to the concentration of tested fractions. The antimicrobial activity of tea leaves was different by collection dates, the quantitative difference was not big. The MIC value of ethyl acetate fraction of tea leaves for Bacillus cereus and B. subtilis was 0.5 mg/ml, on the other hand, the MIC value of the water fraction of tea leaves for B. subtilis, MRSA, Pseudomonas fluorescens and Salmonella typhimurium was 30.0 mg/ml, regardless of collection date. The antimicrobial properties of C. sinensis leaves encouraged an innovative potential application in agricultural, pharmaceutical and food fields.

Potential of The Tyrosinase Enzyme Inhibition by Standardized Ethanol Extract And Ethyl Acetate Fraction of Bengkoang Peel (Pachyrhizus erosus L.)

This study examined the tyrosinase inhibitory potential of bengkoang peel standardised ethanolic extract and ethyl acetate fractions. 450 g of bengkoang peel powder (Pachyrhizus erosus L.) was macerated with 96% ethanol for 2 x 24 h, filtered, and concentrated till viscous. Extract standardisation uses specific and non-specific parameters. Microplate colorimetric tyrosinase inhibition assays were performed. The result of plant determination identified the bengkoang peel as Pachyrhizus erosus (L.) Urb. The organoleptic test revealed a brownish-yellow viscous extract, fresh smell, and bitter taste. Ethanol-soluble extract was 47.60% and water-soluble 22.42%. Bengkoang peel ethanolic extract includes alkaloids, flavonoids, saponins, tannins, and steroids. The non-specific parameters showed loss of drying (9.71%), water content (7.28%), total ash (9.91%), acid-insoluble ash (3.68%), no residual solvent content, lead content (-0.2324±0.1729), cadmium content (-0.0841±0.3418), TPC of 1 x 101 CFU/g, and TYMC of 2.26 x 102 CFU/g. The ethanol extract (103.9263) and ethyl acetate fraction (81.8606) IC50 values of bengkoang peel decrease tyrosinase enzyme activity compared to control (25.1235 ppm). These results show that the ethanol extract met both particular and non-specific quality standards. Bengkoang peel ethyl acetate inhibited tyrosinase.

Antiproliferative Effect of Polysaccharide Fraction of Astragalus Ovinus Against MCF-7 Cancer Cell Line

Background: The effect of Astragalus has been recognized in traditional Chinese medicine, and the roots are believed to have anti-cancer properties. This study investigated the effects of the ethanolic extract and polysaccharide fraction of Astragalus ovinus roots on MCF7 cell line. Methods: We used MTT assay to evaluate the cytotoxicity of A. ovinus roots. The status of cell cycle and apoptosis were examined, using flow cytometry. The gene expressions related to the extrinsic and intrinsic apoptotic pathways (caspases 8 & 9) were also examined by real-time polymerase chain reaction (PCR). Results: The cytotoxicity data showed that the A. ovinus extract inhibited the proliferation of MCF7 cancer cells in a dose-dependent manner. The IC50 of Cisplatin , ethanolic extract and polysaccharides fraction were 22.42, 560.9, and 961.2 in µg/ml, respectively. Also, the cell cycle analysis showed that the ethanolic extract and polysaccharide fractions arrested the cell cycle in the G1 phase. Examination of the apoptotic effects showed that treatment with either the A. ovinus extract or the polysaccharide fraction induced apoptosis in MCF-7 cancer cells. The expression of caspases 8 and 9 genes was inhibited after exposure to either cisplatin or the polysaccharide fraction (PFA). However, treatment with the extract inhibited only the caspase-8 gene expression. Conclusion: The results confirmed that treatment with the extract and polysaccharide fraction caused anti-proliferative effect and apoptosis of MCF-7 cell line. Therefore, it can be concluded that the A. ovinus extract and the polysaccharides have potential therapeutic effects against human breast cancer cells.

Anticholinesterase and anti-inflammatory constituents from Caralluma awdeliana, a medicinal plant from Yemen.

From the dichloromethane (DCM) fraction of the crude ethanolic extract of Caralluma awdeliana, four pregnane glycosides and a flavone glycoside were isolated using a bio-guided isolation approach. The different extracts of C. awdeliana were subjected to in vitro enzyme inhibitory assays of anticholinesterases (AChE and BChE) and anti-inflammatory (COXs and 5-LOX). The highest inhibitory activity was exhibited by DCM fraction against COX-1, COX-2, and 5-LOX with IC50 of 4.8±0.5μg/mL, 0.68±0.2μg/mL, and 39.5±3.0μg/mL, respectively. The DCM showed also a moderate activity against AChE (IC50 384.72±3.6μg/mL), and BChE (IC50 384.72±3.6μg/mL). The repeated chromatography of DCM fraction resulted in the isolation of two new pregnane glycosides, namely awdeliosides A (1) and B (4), two known ones, namely caratuberosides B and D, along with the known flavone glycoside identified as luteolin 4 -O-neohesperidoside. All the isolated compounds were tested for their in vitro enzyme inhibitory assays. Among the isolated compounds, awdelioside B (4) showed the most potent effect against COX-1 with IC50 value of 10.99±0.35μM, compared to standard celecoxib (IC50 230.74±2.62μM). All the isolated compounds showed weak anticholinesterase, except a moderate activity observed for awdelioside B (4) against BChE with IC50 value of 15.63±3.5μM, compared to standard donepezil (IC50 0.77±0.0088μM).

Anticancer and antioxidant activities of ethanolic extract and semi-purified fractions from guava and mango seeds

Mango (Mangifera indica) and guava (Psidium guajava) seeds have several pharmaceutical applications and biological activities because as they have been recognized with different bioactive molecules (phenolic compounds) such as flavonoids, phenolic acids, and catechins, so they have antioxidant and anticancer activities. The aim of the present study was to assess in vitro antioxidant and anticancer activities of successive extracts and semi-purified fractions from mango seeds. In this work, mango and guava seeds were collected and extracted using two solvents (ethanol 70% and ethyl acetate) followed by phytochemical screening and determination of biological activities such as antioxidant activity using five assays (DPPH, ABTS, KMnO4, Methylene blue and H2O2) additionally the antiradical activity and hybrid reaction for ethanolic extract of mango seeds as promising extract. The total phenolic, flavonoid, and catechin compounds were determined for all successive extracts, and finally, the anticancer activity of extracts was evaluated using MTT assay against HepG2 cell line and phenolic compounds were identified by HPLC. The phytochemical screening and TLC showed the primary investigation for phenolic compounds of ethanol extracts of both kind of seeds and only ethyl acetate of guava extract as promising extracts. However, HPLC determination of these three extracts showed high amount of gallic acid, naringenin, ellagic acid, and ferulic acid as they have anticancer and antioxidant activities. The antioxidant tests showed that the ethanolic mango extract is the highest antioxidant extract against DPPH by 84.0%, but recorded 82.0% with methylene blue and ABTS assays when compared with ascorbic acid. The ethyl acetate of guava extract showed strong cytotoxic effect with IC50 75.5 μg/mL against HepG2 cell line in all tested concentrations. From the obtained results, it could be concluded that mango ethanolic extract and its fractions are the most promising as antioxidants and ethyl acetate of guava extract the most promising in the anticancer activity.

Immunomodulatory effect from ethanol extract and ethyl acetate fraction of Curcuma heyneana Valeton and Zijp: Transient receptor vanilloid protein approach

This study aims to discover the immunomodulatory potential of the ethanol extract (EE) and the ethyl acetate fraction (EAF) of Curcuma heyneana Valeton and Zijp (Indonesian name: temu giring) rhizome using mice models. The affinity of the curcuminoid (curcumin, dimethoxy-, and bisdemethoxy-) through the Transient Receptor Potential Vanilloid 1 (TRPV1) was determined using Mollegro molecular docking in silico. The curcuminoid concentration of the EE and EAF of C. heyneana rhizome were determined using thin-layer chromatography densitometry. In vivo studies in mice models were conducted using the carbon clearance method to determine the phagocytosis index, and the number of leukocytes in the blood and spleen. Forty mice were divided into eight groups, including negative control (given 1% CMC-Na), positive control (given Stimuno Forte® suspension at a dose of 6.5 mg/kg BW), three groups given the EAF of C. heyneana rhizome extract at a dose of 125 mg/kg BW, 250 mg/kg BW, and 500 mg/kg BW, respectively, and three groups were given EE of temu giring rhizome extract with doses of 125 mg/kg BW, 250 mg/kg BW, and 500 mg/kg BW, respectively. E.E. and E.A.F. of C. heyneana (temu giring) rhizome extract contained dimethoxy curcumin (0.176 ± 0.01 and 4.53 ± 0.02 %b/b) greater than another curcuminoid, bisdemetoxy curcumin and curcumin. EE at 125 mg/kg BW and EAF dose at 500 mg/kg B W. of temu giring rhizome have immunostimulant activity with a phagocytosis index value of >1 compared to the negative control (p < 0.05). Additionally, both increase the number of lymphocytes, monocytes, and neutrophil cells in peripheral blood and spleen compared to the negative control (p < 0.05). Their activity was seen as similar to the positive control. Therefore, the EE of C. heyneana rhizome has immunostimulant activity, and the EAF of C. heyneana rhizome has immunosuppressant activity at 125 mg/kg BW and immunostimulant at a higher dose. The activity of temu giring as an immunomodulator was associataed with its affinity to TRPV1.

ANTIBACTERIAL AND ANTIACNE ACTIVITY OF n-HEXANE FRACTION OF ALFALFA HERB (Medicago sativa L.) ETHANOL EXTRACT IN VITRO AND ON RABBIT (Oryctolagus cuniculus)

Propionibacterium acnes (P. acnes) can cause inflammation of the polysebaceous glands and lead to acne. Flavonoids in alfalfa herb are thought to have antibacterial and antiinflammatory properties. Flavonoids can be absorbed into n-hexane solvents. This study aims to determine antibacterial activity of n-hexane fraction of alfalfa herb ethanol extract (nHFAH) against P. acnes in vitro, to calculate the total flavonoid levels in the fraction, and to determine antiacne activity of the fraction on rabbit. This study is an experimental study with post-test control group design. Extraction of alfalfa herb was done by maceration method using 70% ethanol and fractionated with n-hexane. Antibacterial activity test was carried out using disk diffusion with 5 series of concentrations (15%, 20%, 25%, 30% dan 35%), positive control (clindamycin 2 µg/disk) and negative control (DMSO 100%). Diameters of the inhibition area were analyzed statistically using a Kruskal Wallis test followed by Mann Withney test with 95% of confidence level. Rabbit back skin was shaved on 5 different areas (3x3 cm2/area). Induced with 0.2 mL of P. acnes suspension (3x109 CFU/mL) intradermal. Acne formed (24 hours) on all areas, then were treated with T1 (clindamycin 2%), T2 (DMSO 100%), T3 (nHFAH 20%), T4 (nHFAH 25%), T5 (nHFAH 30%). Erythema diameter reduction on day 9 was analyzed using one-way anova, continued by post hoc tukey with 95% of confidence level. Total nHFAH flavonoid levels were tested by colorimetric method using UVVis spectrophotometer at λ 429.5 nm. The results showed that nHFAH has antibacterial activity against P. acnes with an average value of diameter of inhibition area for each concentration were 7.30; 7.49; 7.75; 7.92 and 8.24 mm. nHFAH exhibits antiacne activity with the percentage for each concentration were 67.01%; 77.84% and 85.84% respectively. Total nHFAH flavonoid levels were 6.09 mgQE/ gram.Keywords: Alfafa herb, n-Hexane fraction, Propionibacterium acnes.

Total Fenolik, Flavonoid, dan Aktivitas Antioksidan Ekstrak dan Fraksi Eceng Gondok (Eichhornia crassipes (Mart.) Solms)

Water hyacinth is a very fast growing aquatic weed. It contains active compounds including flavonoids, alkaloids, steroids, terpenoids, anthocyanins, tannins, phenolics, anthraquinones and quinones. These compounds can be used as antioxidant, antibacterial, antifungal, antitumor, anticancer, anticoronary and antiinflammatory agents. The purpose of this study was to determine the levels of phenolic and flavonoid in water hyacinth plant extracts and fractions and the IC50 value which describes the level of antioxidant effectiveness. The extract was obtained by maceration using ethanol as a solvent. Fractionation of water hyacinth ethanol extract using n-hexane, ethyl acetate and water as solvents. Antioxidant test used DPPH method. Total flavonoid on the ethanol extract were 0.023±0.0015gEQ/g, the ethyl acetate fraction 0.028±0.0015gEQ/g, the n-hexane fraction 0.008±0.0005gEQ/g, and the water fraction 0.002±0.0005gEQ/g. Total phenolic of water hyacinth herb in ethanol extract were 5.403±0.910mgGAE/g, ethyl acetate fraction was 1.810±0.225mgGAE/g, n-hexane fraction was 0.457±0.290mgGAE/g and water fraction was 0.147±0.151mgGAE/g. The IC50 value for the ethanol extract was 48.64mg/L, the ethyl acetate fraction was 223.27mg/L, the water fraction was 451.86mg/L, and the n-hexane fraction was 845.00mg/L.