Activity Of Ethyl Acetate Fraction Research Articles

Anticancer Activity of Ethyl Acetate Fraction and Ethanol Leaf Extract of Olax subscorpioidea against DMBA-Induced Female Rats

Anticancer Activity of Ethyl Acetate Fraction and Ethanol Leaf Extract of Olax subscorpioidea against DMBA-Induced Female Rats

Evaluation of the anxiolytic activity of ethyl acetate fraction of Calotropis gigantea leaves in rats

Evaluation of the anxiolytic activity of ethyl acetate fraction of Calotropis gigantea leaves in rats

Free radical scavenging polyphenols isolated from Phyllanthus niruri L. ameliorates hyperglycemia via SIRT1 induction and GLUT4 translocation in in vitro and in vivo models

Type 2 diabetes milletus (T2DM) is a complex multifaceted disorder characterized by insulin resistance in skeletal muscle. Phyllanthus niruri L. is well reported sub-tropical therapeutically beneficial ayurvedic medicinal plant from Euphorbiaceae family used in various body ailments such as metabolic disorder including diabetes. The present study emphasizes on the therapeutic potential of Phyllanthus niruri L. and its phytochemical(s) against insulin resistance conditions and impaired antioxidant activity thereby aiding as an anti-hyperglycemic agent in targeting T2DM. Three compounds were isolated from the most active ethyl acetate fraction namely compound 1 as 1-O-galloyl-6-O-luteoyl-β-D-glucoside, compound 2 as brevifolincarboxylic acid and compound 3 as ricinoleic acid. Compounds 1 and 2, the two polyphenols enhanced the uptake of glucose and inhibited ROS levels in palmitate induced C2C12 myotubes. PNEAF showed the potent enhancement of glucose uptake in palmitate-induced insulin resistance condition in C2C12 myotubes and significant ROS inhibition was observed in skeletal muscle cell line. PNEAF treated IR C2C12 myotubes and STZ induced Wistar rats elevated SIRT1, PGC1-α signaling cascade through phosphorylation of AMPK and GLUT4 translocation resulting in insulin sensitization. Our study revealed an insight into the efficacy of marker compounds isolated from P. niruri and its enriched ethyl acetate fraction as ROS scavenging agent and helps in attenuating insulin resistance condition in C2C12 myotubes as well as in STZ induced Wistar rat by restoring glucose metabolism. Overall, this study can provide prospects for the marker-assisted development of P. niruri as a phytopharmaceutical drug for the insulin resistance related diabetic complications.

Antibacterial and Anti Diabetic Activities of Ethyl Acetate Fractions of Lactobacillus Isolated from Cow and Goat Milk

ABSTRACT: Antibiotics are one of our most important weapons in fighting with bacterial infections and diabetics. Drugs were derived from natural sources that play an important role in the treatment and prevention of human diseases. Traditional fermented foods containing lactic acid bacteria such as milk, yogurt, curd, etc., have historically been consumed by human. Lactobacilli are well known friendly bacteria for their probiotic activities against pathogens. In this investigation cow and goat milk samples were selected and collected for the study. The gram stain was used for morphological identification of Lactobacillus sp. in the two samples. The two isolated Lactobacillus sp. from goat and cow milk samples were subjected to the extraction of secondary metabolites using the solvent ethyl acetate. The two extracts of Lactobacillus spp. those were isolated from cow and goat milk samples, showed antibacterial activity against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa. The zone of inhibition was assessed by mm. The good results of zones were observed at 1000 and 2000 µg in cow milk and 2000 µg in goat milk extraction. These metabolites were also showed good results in antidiabetic activity using the alpha amylase inhibition activity. The IC50 value has showed moderate activity in the two crude extractions when compared to acarbose standard drug. Therefore, in this investigation the Lactobacillus sp. producing secondary metabolites was good source for the activity of antibacterial and antidiabetic activity.

Phytochemical profile, acute oral toxicity, antioxidant, and antispasmodic effects of ethyl acetate and aqueous residual fractions of Diospyros mespiliformis Hochst. ex A. DC (Ebenaceae) leaves on isolated duodenum of rat

Introduction: Diospyros mespiliformis Hochst. ex A. Rich (Ebenaceae) is a nutritional, artisanal and medicinal plant. It is used in alternative medicine in Burkina Faso for the treatment of conjunctivitis, menorrhagia, dysentery, and especially diarrhea. Aims: Our study aimed to evaluate the chemical profile, the antioxidant and anti-inflammatory activities, the safety of use, and the spasmolytic effects of the fractions obtained from the aqueous decoction of the leaves of Diospyros mespiliformis. Methods: Phytochemical screening by HPTLC and the determination of compounds of interest were carried out. The antioxidant activity was evaluated according to the ABTS, DPPH, FRAP, and LPO methods. The inhibitory activity of Phospholipase A2 and 15-lipoxygenase was evaluated. Acute oral toxicity was carried out on female mice (NMRI). The ex vivo spasmolytic effect of the fractions was tested on isolated rat duodenum using ACh and BaCl2 as contracting agents. Results: At the end of these tests, the fractions contain flavonoids, tannins, sterols, triterpenes, and saponosides. The content of total phenolics was respectively for the ethyl acetate fraction (EAF) and the residual aqueous fraction (RAF) 84.15±1.73 mg EAT/g and 89.67±2.35 mg EAT/g. That of flavonoids was respectively 45.91 ± 0.98 mg EQ/g and 10.46±0.28 mg EQ/g for the two fractions. The 50% inhibitory concentrations (IC50) of EAF for the ABTS, DPPH, and FRAP tests were 2.26±0.16 μg/mL, 22.34±7.23 μg/mL and 1136.25±0.90 mol EAA/g respectively, and an inhibition % of 43.80 ± 6.31% for LPO. For RAF, the IC50 were in the same range as EAF of 16.82±0.23 μg/mL, 18.58±2.91 μg/mL, 1138.4±1.27 mol EAA/g and an inhibition of 45.14±10.35% for LPO. The fractions had an inhibitory effect on phospholipase A2 compared to Betamethasone. Finally, the most active EAF caused a spasmolytic effect with Emax of 87.4±15.71% and 90.40±7.84%, respectively, during contractions induced by BaCl2 and ACh. Conclusion: Finally, this work provided scientific data and could justify the use of D. mespiliformis leaves in the treatment of diarrhea.
 Keywords: Diospyros mespiliformis, Leaves, Antioxidants, Anti-inflammatory, Safety of use, Spasmolytic

Uric acid–reducing efficacy of Plectranthus amboinicus (Lour.) Spreng in mice with potassium oxonate-induced hyperuricemia

Plectranthus amboinicus (Lour.) Spreng is a type of Lamiaceae perennial herb that is widely used in folk treatments for respiratory and skin diseases. Notably, research on the hypouricemic effect of P. amboinicus is limited. This study investigated the uric acid–reducing efficacy of P. amboinicus in mice with potassium oxonate-induced hyperuricemia by performing an in vitro xanthine-oxidase-inhibition-activity-guided test of P. amboinicus extract (PAE) and its fractions. The ethyl acetate fraction (EAF) of PAE exhibited optimal efficacy in terms of the phenolic content, DPPH scavenging activity, and xanthine oxidase inhibition. We further demonstrated the uric acid–reducing activity of EAF in a mouse model of potassium bromate–induced hyperuricemia. The results can serve as a useful preclinical reference for researching the gout prevention effects of functional foods.
 Keywords: Plectranthus amboinicus (Lour.) Spreng, hyperuricemia, uric acid–reducing activity

Isolation of Biomolecules from the Leaves of Lecaniodiscus cupanoides (Sapindaceae), a Plant used in Traditional Medicine in Benin

The aim of the present study was to identify the structure of the bioactive molecules in the active ethyl acetate fraction of the hydroethanolic extract of Lecaniodiscus cupanoides (Sapindaceae), a Beninese plant used in the treatment of microbial infections. We prepared the hydroethanolic extract from powdered dried leaves. We fractionated the hydroethanol extract using the liquid-liquid extraction method with solvents of increasing polarity. The active ethyl acetate fraction obtained after bioguided fractionation of the hydroethanol extract on bacterial strains was purified by a series of atmospheric pressure column chromatographic methods coupled with thin layer chromatography. At the end of this purification process, three compounds, including a flavonoid and two fatty acids, were isolated and identified by interpretation of 1H NMR, 13C NMR and mass spectrometry spectra. These were: 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-2,3-dihydrochromen-4-one;(E)-3-(3,4-dihydroxyphenyl)-2-(3-(3,4-dihydroxyphenyl) acryloyloxy) propanoic acid and (E)octadec -9- enoic acid (oleic acid).
 The added value of this work lies in the fact that these three molecules have never before been identified in this plant, and therefore represent a potential avenue for the development of a therapeutic arsenal to combat microbial infections.

Bioactivity-Guided Isolation and Antihypertensive Activity of Citrullus colocynthis Polyphenols in Rats with Genetic Model of Hypertension.

Background and Objectives: Citrullus colocynthis belongs to the Cucurbitaceae family and is a wild medicinal plant used in folk literature to treat various diseases. The purpose of the current study was to explore the antihypertensive and antioxidant potentials of Citrullus colocynthis (CC) polyphenol-rich fractions using a spontaneous hypertensive rat (SHR) model. Materials and Methods: The concentrated aqueous ethanol extract of CC fruit was successively fractioned using solvents of increasing polarity, i.e., hexane, chloroform, ethyl acetate and n-butanol. The obtained extracts were analyzed for total phenolic content (TPC), total flavonoid content (TFC) and total flavonol content (TOF). Moreover, the CC extracts were further evaluated for radical scavenging capacity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays and antioxidant activity using inhibition of linoleic acid peroxidation and determination of reducing potential protocols. The phytochemical components were characterized by HPLC-MWD-ESI-MS in positive ionization mode. Results: The results showed that ethyl acetate fraction (EAF) exhibited a higher content of phenolic compounds in term of TPC (289 mg/g), TFC (7.6 mg/g) and TOF (35.7 mg/g). EAF showed higher antioxidant and DPPH and ABTS scavenging activities with SC50 values of 6.2 and 79.5 µg/mL, respectively. LCMS analysis revealed that twenty polyphenol compounds were identified in the EAF, including phenolic acids and flavonoids, mainly myricetin and quercetin derivatives. The in vivo antihypertensive activity of EAF of CC on SHR revealed that it significantly decreased the mean arterial pressure (MAP), systolic blood pressure (SBP), diastolic blood pressures (DBP) and pulse pressure (PP) as compared to normal and hypertensive control groups. Moreover, EAF of CC significantly reduced the oxidative stress in the animals in a dose-dependent manner by normalizing the levels of superoxide dismutase (SOD), malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide (NOx) and total antioxidant capacity (TAC). Furthermore, the treatment groups, especially the 500 mg of EAF per kg body weight (EA-500) group, significantly (p ≤ 0.05) improved the electrocardiogram (ECG) pattern and pulse wave velocity (PWV). Conclusion: It was concluded that the EAF of CC is a rich source of polyphenols and showed the best antioxidant activity and antihypertensive potential in SHR.

Antiproliferative activity of ethyl acetate fraction of Euphorbia ingens against prostate cancer cell line: An in silico and in vitro analysis

Current prostate cancer (PCa) treatments often lead to severe side effects, prompting the exploration of safer alternatives. Euphorbia ingens is a medicinal plant used for cancer treatment in African communities; however, there is no scientific validation of its anticancer activity. This study therefore evaluated the antiproliferative activity of E. ingens on the DU-145 human PCa cell line. The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) bio-assay was used to determine the antiproliferative activity of E. ingens ethyl acetate-, and water-fractions. Only the ethyl acetate fraction was considered to be active. It inhibited DU-145 cell growth selectively without toxicity to non-cancerous Vero E6 cells. The IC50 was 9.71 ± 0.4 µg/ml and had a selectivity index of 8.26, indicating promising efficacy and cellular safety. Qualitative phytochemical screening identified the presence of phenols, terpenoids, flavonoids, tannins, sterols, and saponins in the ethyl acetate fraction. An additional 18 chemical compounds with potential synergistic roles were identified through gas chromatography-mass spectrometry analysis. Network pharmacology was then employed to predict molecular targets and mechanisms of action for drug-like chemical compounds, implicating key targets such as ESR1, IL6, MMP9, CDK2, MAP2K1, AR, PRKCD, CDK1, CDC25B, and JAK2 with regulation of PI3K/Akt, MAPK, and p53 signaling pathways suggested to be the potential mechanism of action. Further, gene expression analysis of selected targets through reverse transcription-quantitative polymerase chain reaction revealed downregulation of AR and BCL2 levels, along with upregulation of p53 and caspase-3 in fraction-treated DU-145 cells compared to the 0.2 % DMSO-negative control. In conclusion, the findings of this study validate the traditional use of E. ingens in cancer management. The identified drug-like compounds, their targets, and associated signaling pathways could serve as a foundation for developing novel strategies for prostate cancer management. However, we recommend additional in vitro and in vivo studies to further substantiate these findings.

Limosilactobacillus reuterias sustainable biological control agent against toxigenic Fusarium verticillioides.

Corn contamination with Fusarium verticillioides (Sacc.) Nirenberg is a worldwide problem that affects yield and grain quality resulting in severe economic losses and implications for food safety. Control of F. verticillioides is a challenge, but lactic acid bacteria (LAB) has high potential as a biological control agent. In this study, the antifungal effect of Limosilactobacillus reuteri (formerly Lactobacillus reuteri) LR-92 against F. verticillioides 97L was investigated. Cell-free supernatant (CFS) from L. reuteri showed concentration-dependent fungicidal and fungistatic activity against F. verticillioides 97L. The antifungal compounds from CFS showed heat stability and pH dependence, and antifungal activity was not affected by treatment with proteolytic enzymes. High-performance liquid chromatography analysis indicated that L. reuteri LR-92 produces lactic and acetic acids. After liquid-liquid extraction, electrospray ionization mass spectrometry analysis of the active ethyl acetate fraction containing antifungal compounds revealed the production of 3-phenyllactic acid, cyclo-(L-Pro-L-Leu), cyclo-(L-Pro-L-Phe), and cyclo-(L-Phe-trans-4-OH-L-Pro). L. reuteri LR-92 has potential as a biocontrol agent for F. verticillioides and contributes to food safety.

GC-MS chemical profiling, antioxidant, anti-diabetic, and anti-inflammatory activities of ethyl acetate fraction of Spilanthes filicaulis (Schumach. and Thonn.) C.D. Adams leaves: experimental and computational studies.

Introduction: This study aimed to investigate the chemical profile of GC-MS, antioxidant, anti-diabetic, and anti-inflammatory activities of the ethyl acetate fraction of Spilanthes filicaulis leaves (EFSFL) via experimental and computational studies. Methods: After inducing oxidative damage with FeSO4, we treated the tissues with different concentrations of EFSFL. An in-vitro analysis of EFSFL was carried out to determine its potential for antioxidant, anti-diabetic, and anti-inflammatory activities. We also measured the levels of CAT, SOD, GSH, and MDA. Results and discussion: EFSFL exhibited anti-inflammatory properties through membrane stabilizing properties (IC50 = 572.79μg/ml), proteinase inhibition (IC50 = 319.90μg/ml), and inhibition of protein denaturation (IC50 = 409.88μg/ml). Furthermore, EFSFL inhibited α-amylase (IC50 = 169.77μg/ml), α-glucosidase (IC50 = 293.12μg/ml) and DPP-IV (IC50 = 380.94μg/ml) activities, respectively. Our results indicated that induction of tissue damage reduced the levels of GSH, SOD, and CAT activities, and increased MDA levels. However, EFSFL treatment restores these levels to near normal. GC-MS profiling shows that EFSFL contains 13 compounds, with piperine being the most abundant. In silico interaction of the phytoconstituents using molecular and ensembled-based docking revealed strong binding tendencies of two hit compounds to DPP IV (alpha-caryophyllene and piperine with a binding affinity of -7.8 and -7.8Kcal/mol), α-glucosidase (alpha-caryophyllene and piperine with a binding affinity of -9.6 and -8.9Kcal/mol), and to α-amylase (piperine and Benzocycloheptano[2,3,4-I,j]isoquinoline, 4,5,6,6a-tetrahydro-1,9-dihydroxy-2,10-dimethoxy-5-methyl with a binding affinity of -7.8 and -7.9Kcal/mol), respectively. These compounds also presented druggable properties with favorable ADMET. Conclusively, the antioxidant, antidiabetic, and anti-inflammatory activities of EFSFL could be due to the presence of secondary metabolites.

Evaluation of Rhodiola rosea Root Fraction as a Functional Cosmetic Ingredient

To develop functional cosmetic material using <i>Rhodiola rosea</i>, the biological activities such as anti-oxidant, anti-inflammatory, astringent, anti-tyrosinase, and anti-elastase activity were investigated with methanol, <i>n</i>-hexane, chloroform, ethyl acetate, n-butanol, and water fractions of <i>R. rosea</i> Root. DPPH and ABTS radical scavenging activities of ethyl acetate fraction were increased in a dosedependent manner; the activities were higher than other fractions. The total phenolic content of ethyl acetate fraction also showed a relatively high value of 500.485±4.848 mgGAE/g. The ethyl acetate fraction also significantly decreased LPS-induced NO production and IL-6 secretion compared with untreated control cells at above 5-100 μg/mL concentrations. The astringent activity of ethyl acetate fraction was 94.31% at a concentration of 500 μg/mL, and the activity of ethyl acetate fraction was similar to that of tannic acid. The mushroom tyrosinase inhibition abilities of hexane, ethyl acetate, and butanol were 10.44%, 17.53%, and 14.55%, respectively, at 100 μg/mL. Furthermore, an elastase inhibition rate showed a relatively high effect at 100 μg/mL, 45.46% of inhibition rate in the ethyl acetate and 45.07% of inhibition rate in the butanol fraction. These results indicate that R. rosea Root would be used as a natural material in the functional cosmetic and beauty-food industries.

Comparison of Antibacterial Activity of Ethyl Acetate Fractions Separated from Trichoderma with Commercial Antibiotic Methicillin Against Drug-Resistant Bacteria using Minimum Inhibitory Concentration Method

Aim: To compare the antibacterial activity of ethyl acetate fractions of Trichoderma viride against Drug-resistant bacteria. Materials and method: The ethanolic extract was made from fully grown mycelia and Trichoderma was isolated from the soil sample. Four Methicillin-Resistant Staphylococcus aureus strains were examined with the ethyl acetate extract (MRSA). The sample size was calculated using Clincalc.com, which revealed that the alpha error-threshold 0.05, the confidence interval 95%, the g-power 80%, and the enrollment ratio was 1. Groups 1 and 2 with sample size 12 each which represented drug-resistant bacteria and ethyl acetate fractions respectively. Result: The statistical Independent Sample T-Test analysis showed significant antibacterial activity against MRSA for the Trichoderma ethanolic extract with the statistical significance (p<0.05) when compared to the Susceptible bacterial strains. Conclusion: The tested ethyl acetate extract showed the MRSA at 100 μg/ml.

Ciliatoside A, isolated from Peristrophe japonica, inhibits HBsAg expression and cccDNA transcription by inducing autophagy

Hepatitis B surface antigen (HBsAg) loss and seroconversion are considered as an end point of a functional cure. Therefore, it is crucial to find new agents which could efficiently decrease HBsAg. Traditional herbal plants have been considered as an important source of new hepatitis B drugs development for their extensive use in antimicrobial and anti-inflammation. In this study, Peristrophe japonica, which could remarkably reduce HBsAg in the supernatant of HepG2.2.15 cells, was screened out for further extraction. Here, an active ethyl acetate fraction of Peristrophe japonica containing 34 sub-fractions was extracted. Subsequently, the monomeric compound Ciliatoside A was isolated and identified as a potential antiviral reagent with low cytotoxicity from Fraction 30. Ciliatoside A exhibited strong inhibition on intracellular and circulating HBsAg and HBV RNAs in HBV-infected cells and an HBV recombinant-cccDNA mouse model. The mechanistic study revealed that Ciliatoside A exhibited a potent anti-HBV effect through inducing autophagy-lysosomal pathway to autophagic degradation of HBc by activating AMPK-ULK1 axis and inhibiting mTOR activation. In summary, we have identified a novel antiviral compound Ciliatoside A isolated from Peristrophe japonica. This study may provide important direction and new ideas for the discovery of hepatitis B cure drugs.

Bronchodilator Secondary Metabolites from Rhazya stricta Decne Aerial Parts

The plant kingdom comprises medicinally useful plants that have provided many new drugs used to treat various diseases. In our search for bronchodilator secondary metabolites from plants growing in Saudi Arabia, the total extract of Rhazya stricta showed activity against carbamylcholine- (CCh) induced bronchoconstriction in guinea pig tracheal muscles used as an ex vivo model. The fractions obtained from liquid–liquid extraction process were tested for bronchodilator effects. The most active ethyl acetate fraction (RS-E) and aqueous fraction (RS-H) were subjected to biologically guided phytochemical study using different stationary phases and chromatographic techniques to isolate the pure secondary metabolites. Five known compounds were isolated from the active fractions. Three alkaloids namely; (-)-quebrachamine (1), (+)-eburenine (2), (+)-stemmadenine (3) as well as the two iridoid glycosides loganic acid (4) and loganine (5) were identified by various spectroscopic methods. Among the isolated compounds 1 and 5 were the only active as bronchodilators in the plant. It is worth to mention that iridoid glycosides are isolated for the first time from R. stricta.

Total flavonoid content and analgesic activity of ethyl acetate fraction of Indian jujube (Ziziphus mauritiana lamk) leaves

Indian jujube leaves (Ziziphus mauritiana Lamk) have various potential biological activity. Previous studies have proven that ethanol extracted from Indian jujube (Z. mauritiana Lamk) leaves had analgesic activity. This study aimed to determine the total flavonoid content and analgesic activity of the ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves ethanol extract. Tested fraction was prepared by maceration using ethanol as solvent followed by fractionation using ethyl acetate. Identification of active compounds was performed using color reaction, and determination of total flavonoids by colorimetric method. Ethyl acetate fraction was further tested for its analgesic activity using the writhing method. Swiss-Webster white male mice were divided into 5 treatment groups consisting of negative control contained sodium-CMC, positive control contained sodium diclofenac, while groups 3-5 were given ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves with doses of 50, 150, and 200 mg/kg BW. All interventions were administered as a single oral dose. 0.7% (w/v) acetic acid was used as a pain inducer. Analgesic activity was measured by calculating the percentage of writhing protection as a measure of the analgesic effect produced by each intervention. The data were analyzed using one-way Anova to compare analgesic activity between treatment groups. Results showed that ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves at doses of 50, 150, and 200 mg/kg BW gave an effect that was not significantly different from sodium diclofenac as positive control. The ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves ethanol extract contained flavonoids, saponin, and tannins with total flavonoid content of 22.41 ± 1.16% w/w. It can be concluded that the ethyl acetate fraction of Indian jujube (Z. mauritiana Lamk) leaves have analgesic activity in mice.

In Vitro Bioactivities of Extracts from Tomato Pomace.

Tomato pomace (TP) is a coproduct generated by the extraction of tomato pulp, and is a potential source of bioactive molecules. In this study, we isolated several fractions from TP and evaluated their biological properties. TP was treated by maceration at room temperature with green solvents (ethanol, ethyl acetate, ethanol:water and ethanol:ethyl acetate) or supercritical CO2 (SC-CO2). The extracts were analyzed by HPLC-DAD to determine their composition, and their antioxidant activity was assessed. The potential therapeutic effects of the isolated fractions were assessed in vitro. We identified 30 molecules on chromatography profiles, which revealed an abundance in phenolic acids, carotenoids, flavonoids and tannins, with differences in selectivity according to the solvent and pretreatment used. The highest radical scavenging activities were measured at 64-72% inhibition, corresponding to the ethanol or ethanol:water extracts with the highest polyphenol or flavonoid contents. Carotenoid content was increased by chemical pretreatment, to attain levels of 161 mg β-carotene/g ethyl acetate extract. This level of carotenoids seemed to have anti-inflammatory effects, with an IC50 of 9.3 μg/mL. In terms of anti-diabetic effects, the activities of α-glucosidase and α-amylase were best inhibited by extraction in an ethanol-to-water mixture (50:50). Cytotoxicity in a tumor cell line were highest for SC-CO2 extracts (64.5% inhibition) and for ethanol extracts obtained after the enzymatic pretreatment of TP (37% inhibition). Some extracts also had dose-dependent activity against Zika virus. New fractions obtained from TP with ecocompatible solvents in mild conditions are rich in bioactive molecules. A comparison of the chromatographic profiles of the extracts led to the identification of several key molecules with therapeutic properties. The chemical pretreatment of TP is justified as a mean of increasing the carotenoid content of ethyl acetate fractions, whereas enzymatic pretreatment can increase the antioxidant activity of ethyl acetate fractions and increase the cytotoxicity of ethanol fractions. The SC-CO2 fraction contained a smaller number of metabolites detectable on HPLC, but it had high levels of cytotoxicity and antioxidant activity. Finally, the fractions obtained appeared to be suitable for use to target one or several of the biological activities studied.

Determination of Total Flavonoid Level and Antioxidant Activity of Ethyl Acetate Fraction of Mangkokan Leaf Extract (Nothopanax scutellarium [Burm.f] Merr.)

​Mangkokan leaves (Nothopanax scutellarium (Burm.f) Merr.) have several properties, containing several chemicals that have bioactivity. It contains flavonoids and is a natural source of antioxidants. This study aims to determine the total flavonoid content and the potency of the ethyl acetate fraction in the leaves of the Mangkokan leaf as an antioxidant in counteracting free radicals. Identification of flavonoids was carried out with a reagent between magnesium powder and concentrated HCl, the orange color formed indicated the presence of flavonoids. Determination of the total flavonoid content of the ethyl acetate fraction of the Mangkokan leaf extract was carried out colorimetrically using a UV-Vis spectrophotometer. The antioxidant activity test of the ethyl acetate fraction of kukukan leaf extract was carried out using the DPPH (1,1-diphenyl-2-picrylhydrazyl) method. The results showed that the total flavonoid content of the ethyl acetate fraction of the Mangkokan leaf extract was 25.402749 mgGAE/g extract and the free radical inhibition test with the DPPH method showed that the ethyl acetate fraction of the Mangkokan leaf extract had antioxidant activity with an IC50 value of 103.36 ppm. The ethyl acetate fraction of the Mangkokan leaf extract has activity as an antioxidant with a moderate category of antioxidant power so that it has the potential to be developed in the treatment of diseases caused by free radicals.

Melissa officinalis L. nanoethosomal formulation: evaluation of antioxidant, enzyme inhibitory activities and in vitro toxicity

ABSTRACT This work aimed to create an extract of Melissa officinalis L. with strong antiradical efficacy, characterize it, and enhance its long-term efficacy by developing an ethosomal formulation. DPPH and ABTS assays were used to test the antiradical activity of extracts with different ethanol ratios obtained from the aerial part. Phytochemical characterization of the extract with the highest activity, ethyl acetate fraction of 60% ethanol extract, was analyzed by HPLC. The active ethyl acetate fraction was loaded into ethosomes, and characterization and release studies of the formulation were performed. The released extract from the formulation exhibited substantial antiradical action as well as inhibition of collagenase (71.5%) and elastase (75.5%) enzymes. The toxicity of the active extract and the formulation was determined in the mouse fibroblast cell line. This study successfully developed a long-term antioxidant and enzyme inhibitor formulation containing M. officinalis, which stands out for its medicinal properties.

Anticancer and Neuroprotective Activities of Ethyl Acetate Fractions from Morus macroura Miq. Plant Organs with Ultraperformance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry Profiling.

Column chromatography afforded the isolation of seven secondary metabolites (1-(2,4,6-trihydroxy phenyl)-ethanone-4-O-β-d-glucopyranoside, naringenin-7-O-β-d-glucopyranoside, kaempferol-3-O-α-l-rhamnoside, kaempferol-3-O-β-d-glucopyranoside, quercetin-3-O-β-d-glucopyranoside, quercetin-3-O-β-d-galactopyranoside, rutin) from the ethyl acetate (ET) fractions of Morus macroura Miq. stems (S), leaves (L), and fruits (F). Their identification based on ultraviolet (UV), electron ionization (EI), electrospray ionization-mass spectrometry (ESI-MS), and 1D and 2D NMR data. In addition, profiling of ET fractions using ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) resulted in the identification of 82 compounds belonging to different classes, mainly polyphenolic constituents. Chemical profiling as well as molecular docking directed us to biological evaluation. Interestingly, the ET-L fraction exhibited a robust cytotoxic activity against HepG-2, MCF-7, and HELA cell lines. Also, it displayed a neuromodulatory activity against cisplatin neurotoxicity in rats by ameliorating the neurobehavioral dysfunction visualized in the open field and Y-maze test and modulating the neurochemical parameters such as brain amino acid levels (glutamate, aspartate, serine, and histidine), oxidative stress markers (GSH, MDA, and 8-hydroxy-2′-deoxyguanosine), and purinergic cell energy (adenosine triphosphate (ATP) and adenosine monophosphate (AMP)). In conclusion, the isolated compounds (kaempferol-3-O-β-glucoside and quercetin-3-O-β-glucoside) from the ET-L fraction could serve as potent anticancer agents due to their strong antioxidant, in vitro cytotoxicity, and in vivo neuroprotective activity.