EtOAc Fraction Research Articles

Antioxidant and Anti-inflammatory Activities of the Extract and Solvent Fractions of Hemistepta lyrata Bunge

This study investigated antioxidant and anti-inflammatory in order to confirm the possibility of functional materials that can improve skin diseases using <i>Hemistepta lyrata</i> Bunge extracts and solvent fractions. As a result of measuring the total polyphenol content, the content was high in EtOAc fraction, and then the BuOH fraction is next. As a result of measuring DPPH free radical scavenging activities, it was confirmed that BuOH fractions had superior antioxidant activity compared to other solvent fractions. In particular, in the case of EtOAc fraction, it was confirmed that the antioxidant ability was excellent even at a low concentration compared to other fractions in ABTS and FRAP assay. Through this, it was confirmed that the higher the total polyphenol content, the higher the antioxidant effect. In addition, as a result of verifying the inflammatory efficacy of the extract and the solvent fraction using the J774 cell line, it was confirmed that the NO level decreased when the extract and the solvent fraction were treated. In particular, it is determined that the ethyl acetate fraction having a high total polyphenol content has superior antioxidant and anti-inflammatory activities and contains more active substances than other fractions. Therefore, in order to conduct additional research, it is determined that research on the compounds contained in the ethyl acetate fraction is necessary.

Revisiting the Flora of Saudi Arabia: Phytochemical and Biological Investigation of the Endangered Plant Species Euphorbia saudiarabica.

Euphorbia plants have a significant place in traditional medicine due to their numerous therapeutic properties, including their anti-tumor effects, which have been observed in several species. In the current study, a phytochemical investigation of Euphorbia saudiarabica methanolic extract led to the isolation and characterization of four secondary metabolites from the chloroform (CHCl3) and ethyl acetate (EtOAc) fractions, which are reported for the first time in this species. One of the constituents, saudiarabicain F (2), is a rare C-19 oxidized ingol-type diterpenoid that has not been previously reported. The structures of these compounds were determined by extensive spectroscopic (HR-ESI-MS, 1D and 2D NMR) analyses. The anticancer properties of the E. saudiarabica crude extract, its fractions and its isolated compounds were examined against several cancer cells. The active fractions were evaluated for their effects on cell-cycle progression and apoptosis induction using flow cytometry. Furthermore, RT-PCR was employed to estimate the gene-expression levels of the apoptosis-related genes. It was demonstrated that the E. saudiarabica CHCl3 and EtOAc fractions suppressed the proliferation of the cancer cells. The MCF-7 cells were the most sensitive to both fractions, with IC50 values of 22.6 and 23.2 µg/mL, respectively. Notably, both fractions caused cell-cycle arrest in the G2/M phase of the treated MCF-7 cells. The inhibition of the MCF-7 cells' proliferation was also linked with apoptosis induction by flow-cytometry analysis. Additionally, the activation of apoptosis by both fractions was demonstrated by an increase in the ratio of Bax to Bcl-2, with an increase in the expression of caspase-7. Among the isolated compounds, glutinol (1) showed potent activity against the MCF-7 cell line, with an IC50 value of 9.83 µg/mL. Our findings suggest that E. saudiarabica has apoptosis-inducing effects and shows promise as a potential source of new chemotherapeutic drugs.

Phytochemicals and Antioxidant Activities of Red Oak, Red Coral and Butterhead.

Lactuca sativa L. is an economically important vegetable that contains numerous phytochemicals. This study aimed to determine the phytochemicals in three lettuce cultivars (red oak, red coral, and butterhead) and assess their total phenolics, total flavonoids and antioxidant activities. The dried leaves of each lettuce cultivar were macerated with hexane, ethyl acetate (EtOAc), and 95% ethanol (EtOH). Total phenolics, total flavonoids, and antioxidant activities from the three solvent extracts were measured. The phytochemical screening showed that the leaves from the three lettuce cultivars contained flavonoids, hydrolyzable tannins, coumarins, steroids, and phenolic compounds. While the EtOAc fraction of the red coral lettuce showed the highest total phenolic content (9.747 ± 0.021 mg gallic acid equivalent/g) and the hexane fraction of the butterhead lettuce contained the highest flavonoids (7.065 ± 0.005 mg quercetin equivalent/g). In the DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay, the EtOAc fraction of the red coral lettuce had the highest antioxidant capacity with an IC50 of 0.277 ± 0.006 mg/mL, whereas, in the ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assay, the 95% EtOH of the red coral lettuce had the highest antioxidant capacity with an IC50 of 0.300 ± 0.002 mg/mL. All three lettuce cultivars contained high levels of phenolic content and flavonoids, which are the source of antioxidant activities. These lettuce cultivars, especially the red coral lettuce, are a potential source of natural antioxidants. Further research on the application of natural antioxidants is required to investigate the therapeutic or the neutraceutical implication of the lettuce cultivars.

Flavonol-Glycoside and Rare Triterpenoid Derivatives Isolated from Leaves of Combretum glutinosum Perr. Ex Dc. with In Vitro Cytotoxic Activity

Combretaceae plants are used traditionally by many cultures, especially in Sudanese patients for the treatment of diverse ailments such as anti-inflammatory, antimicrobial, antitumor, and antioxidant disorders. Of these plants, the genus Combretum are traditional medicinal plants. Thus, they are formed from the non-polar or polar extracts of many isolated phytochemicals. Of these necessities, the use of Combretum extracts for their medicinal properties can be found in the earliest of myths and traditions used to document the plants’ ability to treat diseases. Combretum glutinosum Perr. Ex Dc. is a common shrub native to the African continent, especially Sudan. Currently, there are no published data regarding its cytotoxic activity. Additionally, there are few chemical and biological reports of C. glutinosum. Therefore, the current study aimed to isolate the chemical bioactive compounds (1–6) from the ethyl acetate (EtOAc) extract of C. glutinosum. A new flavonoid compound, namely, glutosinumoside (4), was afforded, and five known compounds were obtained: three oleanane-glycosides (1–3) and two phenolic acids (5,6). The structures of the six compounds were determined by spectroscopic analysis, including one- and two-dimensional (1D and 2D) NMR, mass spectrometry, and chromatographic analysis. Moreover, an in vitro cytotoxic evaluation of the successive extracts and the bioactive EtOAc fractions of C. glutinosum against MCF7 (breast), HT29 (colon), HepG2 (liver), and MRC5 (normal lung) cell lines was performed. The isolated compounds showed comparable cytotoxic activities with the crude EtOH extract and doxorubicin against the tested cell lines. Compounds (1) and (6) exhibited the highest cytotoxicity against MCF7 (1.37 ± 0.21 and 1.48 ± 0.34 µg/mL, respectively) and HepG2 (3.30 ± 0.02 and 2.10 ± 0.22 µg/mL, respectively) in the MTT assay. In addition, compounds (1) and (3) demonstrated a significant upregulation of cancer’s two important hallmarks (caspase 3 and bax genes) by inducing apoptosis and perturbing the MCF7 cell cycle.

Identification of the Constituents of Ethyl Acetate Fraction from Smilax china L. and Determination of Xanthine Oxidase Inhibitory Properties

The aim of this work was to investigate the xanthine oxidase (XO)-inhibitory activity of ethanol extracts from Smilax china L. and to identify the active compounds in the ethyl acetate (EtOAc) fraction. Extraction of ethanol extracts from Smilax china L. and then ethanol extracts were concentrated, and the polyphenolic compounds were extracted with petroleum ether (PE), chloroform, EtOAc, n-butanol (n-BuOH), and residual ethanol fractions. Their effects on XO activity were then compared separately. The polyphenolic components of the EtOAc fraction were identified by HPLC and HPLC-mass spectrometry (HPLC-MS) analysis. Kinetic analysis demonstrated that all these extracts showed XO-inhibitory properties, and among them the EtOAc fraction had the strongest inhibitory effect (IC50 = 101.04 μg/mL). The inhibitory constant (Ki) of the EtOAc fraction on XO activity was 65.20 μg/mL, showing excellent inhibition on XO in the competitive mode. Sixteen compounds were identified from the EtOAc fraction. The study demonstrates that the EtOAc fraction of Smilax china L. may be a potential functional food to inhibit XO activity.

Desmoarylcoumarin and episoyasapogenol B: two new compounds from Desmodium salicifolium (Poir.) DC (Fabaceae)

A new 3-arylcoumarin, 7-hydroxy-6-(1,1-dimethylallyl)-2′,5′-dihydroxy-4′-(3,3dimethylprenyl)-3-arylcoumarin (desmoarylcoumarin) 1, a previously unreported oleanane-type triterpenoid, 3β,22β,23-trihydroxyolean-12-en (episoyasapogenol B) 2, together with five known flavonoids including darbergioidin (3), isoferreirin (4), quercetin (5), vitexin (6), swertizin (7), and one carbohydrate, sucrose (8) were isolated from the methanolic extract of the roots of Desmodium salicifolium. Their structures were elucidated mainly by extensive spectroscopic analysis (1D and 2D) and mass spectrometric (HRFAB-MS) data. The methanolic extract, EtOAc and n-BuOH fractions as well as some isolated compounds were assessed for their antibacterial and antioxidant activities. The EtOAc fraction exhibited moderate activity against Enterococcus faecalis with MIC value of 128 µg/mL. The methanolic extract and the EtOAc fraction displayed DPPH scavenging activity with EC50 values of 5.99 and 2.06 μg/mL, respectively. Compound 1 showed a moderate antibacterial activity against Enterococcus faecalis with a MIC of 16 µg/mL. It also showed moderate DPPH scavenging activity.

In vitro immunomodulatory activity study of Garcinia cowa Roxb. fraction.

The objective of this study is to determine the activity of Garcinia cowa Roxb. n-hexane, ethyl acetate, and butanol fractions as an immunomodulator invitro and obtain the fraction that has the potential as an immunomodulator. Raw 264.7 macrophages were used to asses G.cowa Roxb. immunomodulatory activity. The MTT assay was chosen to measure cell viability to evaluate the cytotoxic effect on cells. ELISA method was used to measure the concentration of Interleukin-6 (IL-6) and Tumor Necrosis Factor Alpha (TNF-α) secreted by cells after being treated with G.cowa Roxb. fraction. The neutral red uptake assay determined the effect of Garcinia cowa Roxb. on the phagocytic activity. After Raw 264.7 macrophages were given the Hexan fraction (Hex) at concentrations of 12.5 and 25μg/mL, there was a decrease in the concentration of IL-6, TNF-α, and the phagocytosis index of cells. Administration of the Ethyl Acetate fraction (EtOAc) at concentrations of 12.5 and 25μg/mL on cells caused a decrease in IL-6 and TNF-α levels but did not affect the phagocytosis index. There was an increase in the level of TNF-α and the phagocytosis index after being given the Butanol fraction (BuOH) with concentrations of 12.5 and 25μg/mL but there was a slight decrease in the level of IL-6. Both Hex and EtOAc fractions could suppress immune responses through decreasing IL-6, TNF-α, and slightly decreased phagocytic activity. BuOH fraction could stimulate immunomodulatory activities through enhanced TNF-α levels and phagocytic index, but less potent in enhancing IL-6 production. The BuOH fraction could be developed as an immunostimulant.

Chemical constituents and in vivo preliminary evaluation of the toxicological activity of Ouratea spectabilis (OCHNACEAE) and Clitoria guianensis (Fabaceae) leaves

Clitoria guianensis and Ouratea spectabilis, found in the Brazilian Cerrado, are used in folk medicine, despite the few chemical and biological studies reported in the literature. The present study aims to investigate the toxicity and effect of extracts from both species on the microcrustacean Artemia salina, and to determine the chemical composition of the hexane extract of O. spectabilis leaves and the EtOAc fraction of C. guianensis leaves. Kaempferitrin, a flavonoid isolated from of the EtOAc fraction of C. guianensis leaves, was identified by chemical analysis. Analysis of the hexane extract of O. spectabilis leaves using gas chromatography-mass spectrometry (GC-MS) suggested the presence of twenty-five known substances. The Hex, EtOAc, and EtOH crude extracts of C. guianensis leaves exhibited high and moderate toxicity against Artemia salina, with median lethal dose values (LD50) of 43.7, 25.4, and 233.4 mg.L−1, respectively. The acetone extract of O. spectabilis leaves showed moderate toxicity against Artemia salina with an LD50 value of 115.13 mg.L−1.

Antioxidant and antibacterial activities of the rhizome extract of Curcuma zedoaria extracted using some organic solvents.

This research aims to identify the effect of various organic solvents such as n-hexane, ethyl acetate (EtOAc), and methanol (MeOH) on the antioxidant and antibacterial activities of Curcuma zedoaria extract, against three Gram-positive bacteria, namely Staphylococcus aureus, Bacillus subtilis, and Streptococcus pneumoniae, and three Gram-negative bacteria, namely Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa. As much as 1 kg of white turmeric rhizome (C. zedoaria) was extracted two times for 24 h using 3 l of MeOH before evaporating. The extract was then fractionated using n-hexane six times per 2 h, with each volume of 500 ml, and continued with the EtOAc fractionation. The MeOH fraction was added to 300 ml of water before adding 400 ml of EtOAc. Once the fractionation process was complete, all fractions were concentrated using a rotary evaporator. The C. zedoria extract fractioned using MeOH produces alkaloids, phenolics, flavonoids, saponins, and coumarin compounds. The fractionation with EtOAc also produces alkaloids, phenolics, flavonoids, saponins, coumarin compounds, and triterpenoids. Meanwhile, fractionation with n-hexane only produces alkaloids and triterpenoid compounds. EtOAc and MeOH fractions had good activity in reducing free radicals produced by 2,2-diphenyl-1-picrylhydrazyl (DPPH), with an average IC50 value of 153.49 ± 2.66 and 185.77 ± 3.91 ppm, respectively. In contrast, the n-hexane fraction has weak antioxidant activity with an IC50 value of 837.92 ± 5.32 ppm. The n-hexane fraction has better activity compared to MeOH and EtOAc. The lowest concentration required was 2,500 ppm for all types of bacteria. Curcuma zedoaria extract produces alkaloids, phenolics, flavonoids, saponins, coumarins, and triterpenoids when fractionated with MeOH or EtOAc. Only alkaloids and triterpenoids are produced using n-hexane. EtOAc and MeOH fractions have good activity in reducing free radicals generated by DPPH, with an average IC50 value of 153.49 ± 2.66 and 185.77 ± 3.91 ppm, respectively. However, n-hexane has weak antioxidant activity, with an average IC50 value of 837.92 ± 5.32 ppm. All fractions have moderate antibacterial activity, but the extract of n-hexane from C. zedoary has better antibacterial activity compared to MeOH and EtOAc. The lowest concentration required is 2,500 ppm for all types of bacteria.

Triterpenoid saponins and C21 steroidal glycosides from Gymnema tingens and their glucose uptake activities.

Four new triterpenoid saponins, tigensides A-D (1-4), and one new C21 steroid, tipregnane A(9), together with six known compounds were isolated from the EtOAc fraction of the roots and stems of Gymnema tingens. The chemical structures of the new compounds were determined based on their spectroscopic data, including IR, UV, NMR, and mass spectrometric analysis. All compounds were isolated for the first time. Compounds 1-11 promoted glucose uptake in the range of 1.12 to 2.52 fold, respectively. Compound 2 showed the most potent glucose uptake, with 2.52 fold enhancement. Additionally, compound 2 showed a medium effect on the GLUT4 translocation activity in L6 cells in further study.

Antibacterial constituents of Rumex nepalensis spreng and its emodin derivatives

The CH2Cl2−MeOH (1:1) extract of roots of Rumex nepalensis (Polygonaceae) displayed significant antibacterial activity against five bacterial strains with MICs (62.5–31.2 μg.mL−1). The EtOAc soluble fraction displayed a significant activity against the same strains with MICs (31.2–3.9 μg.mL−1). The purification of the EtOAc fraction yielded one new phenylisobenzofuranone derivative, berquaertiide (1), along with 19 known compounds (2–20). Their structures were elucidated based on the analysis of their NMR and MS data. All the isolated compounds were assessed for their antibacterial activity. Compound 2 was the most active against all the tested strains (15.7 to 1.9 μg.mL−1), while compounds 3–7 displayed good activities on at least one of the tested strains. In addition, seven analogues (21–27) of compound 2 were prepared and further assessed for their antibacterial activity. Compounds 26 and 27 were most active than 2 against Salmonella enterica and Klebsiella pneumoniae with MIC (125 and 15.6 μg.mL−1, respectively).

Human Lung Cancer (A549) Cell Line Cytotoxicity and Anti-Leishmania major Activity of Carissa macrocarpa Leaves: A Study Supported by UPLC-ESI-MS/MS Metabolites Profiling and Molecular Docking.

Lung cancer and cutaneous leishmaniasis are critical diseases with a relatively higher incidence in developing countries. In this research, the activity of Carissa macrocarpa leaf hydromethanolic extract and its solvent-fractions (n-hexane, EtOAc, n-butanol, and MeOH) against the lung adenocarcinoma cell line (A549) and Leishmania major was investigated. The MeOH fraction exhibited higher cytotoxic activity (IC50 1.57 ± 0.04 μg/mL) than the standard drug, etoposide (IC50 50.8 ± 3.16 μg/mL). The anti-L. major results revealed strong growth inhibitory effects of the EtOAc fraction against L. major promastigotes (IC50 27.52 ± 0.7 μg/mL) and axenic amastigotes (29.33 ± 4.86% growth inhibition at 100 μg/mL), while the butanol fraction exerted moderate activity against promastigotes (IC50 73.17 ± 1.62), as compared with miltefosine against promastigotes (IC50 6.39 ± 0.29 μg/mL) and sodium stibogluconate against axenic amastigotes (IC50 22.45 ± 2.22 μg/mL). A total of 102 compounds were tentatively identified using UPLC-ESI-MS/MS analysis of the total extract and its fractions. The MeOH fraction was found to contain several flavonoids and flavan-3-ol derivatives with known cytotoxic properties, whereas the EtOAc fractions contained triterpene, hydroxycinnamoyl, sterol, and flavanol derivatives with known antileishmanial activity. Molecular docking of various polyphenolics of the MeOH fraction with HDAC6 and PDK3 enzymes demonstrates high binding affinity of the epicatechin 3-O-β-D-glucopyranoside and catechin-7-O-β-D-glucopyranoside toward HDAC6, and procyanidin C2, procyanidin B5 toward PDK3. These results are promising and encourage the pursuit of preclinical research using C. macrocarpa's MeOH fraction as anti-lung cancer and the EtOAc fraction as an anti-L. major drug candidates.

GC-MS Analysis and Antioxidant Activity of EtOAc Extract Fraction of Acassia sieberiana (Fabaceae) for the Presence of Flavonoids

This research is aimed at analyzing the flavonoid content of ethyl acetate (EtOAc) extract fraction of Acassia sieberiana through Gas chromatography mass spectrometry (GC-MS) and testing its radical scavenging activity on DPPH. The stem bark of the plant was collected from Pankshin local government area of Plateau state. It was made into fine powder and macerated in methanol. The concentrated extract was fractionated in EtOAc and water. The EtOAc fraction was concentrated using rotary evaporator. The concentrated EtOAc extract fraction was subjected into phytochemical screening which revealed the presence of cardiac glycosides, tannins, saponins, flavonoids, phenolic compounds and terpenes. The components of the EtOAc fraction were separated by TLC using solvents mixture of n-hex:EtOAc in the ratio 3:1 and n-hex:EtOAc:MeOH in the ratio 5:1:1. Flavonols, aurones, flavones, flavonones and catechines were indicated. The GC-MS showed the presence of Ergosta–5,22-dien-3-01-acetate(93β,22E), 4H-Benzopyran-4-one,5,6,7-trimethoxy-2-(4-methoxyphenyl), Ethyliso-allocholate, 3,9- epoxypreg-16-en-14-ol-20-one-11,18-diacetoxy-3-methoxy and 10-Octadecnoicacidmethyl ester at retention times 30.038, 31.685, 32.057, 34.148 and 38.780min respectively. The antioxidant activity of the EtOAc extract fraction was appreciable in the sense that it has IC50 of 57.18ug/ml though less than that of ascorbic acid which was found to be 52.04ug/ml.

Chemical constituents of Pteris excelsa and their antioxidant activity

Pteris excelsa Gaud. is a perennial evergreen herbaceous plant belonging to the Pteridaceae family. In the present study, the chemical constituents of this species were obtained by utilizing MCI, C18 and silica gel and Sephadex LH-20 gel column chromatography. A new flavonol glycoside was identified, kaempferol-4′,7-dimethyl-3-O-β-D-glucosyl-(1 → 4)-α-L-rhamnoside (1), together with fourteen known compounds (2–15), kaempferol-4′-methyl-3-O-β-D-glucoside (2), kaempferol-4′,7-dimethyl-3-O-glucoside (3), kaempferol-3-O-β-D-glucoside (4), kaempferol- 4′-methyl-3,7-α-O-L-dirhamnoside (5), kaempferol-3,7-di-O-α-L-rhamnoside (6), pterosin B (7), 3β-hydroxycinnamolide (8), N-(3-carboxypropyl)-2-acetylpyrrole (9), vanillic acid (10), sauropunol C/D (11), 6-anhydro-2-deoxy-β-O-furanosides (12), aquilegiolide (13), menisdaurilide (14), 2-(D-glycerol-1,2-dihydroxyethyl) furan (15). The structures of these compounds were established based on their high-resolution electrospray ionization mass spectrometry results and nuclear magnetic resonance spectroscopy data and by comparing their spectroscopic data with those reported in the literature. Importantly, we found that the antioxidant activity of PE, EtOAc and n-BuOH fractions of 98% methanol extract of P. excelsa (mediated by scavenging free radicals) was 33.57 ± 0.67%, 52.98 ± 1.55% and 45.25 ± 1.10% at a concentration of 400 μg/mL, respectively. Besides, compounds 1–6 displayed an antioxidant effect against the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical.

Antidiabetic and Renoprotective Effects of Ethyl Acetate Fraction of Mimosa pudica Linn. Leaves Extract in Streptozotocin-Induced Diabetic Mice

Background: Traditional medicine has used Mimosa pudica Linn. for diabetes treatment. This study evaluates the antidiabetic and renoprotective effects of the ethyl acetate fraction of M. pudica leaves on streptozotocin-induced diabetes in mice. Methods: The cold maceration method was used to extract M. pudica leaves with 80% ethanol at room temperature. Ethyl acetate (EtOAc) fraction was obtained from the M. pudica leaves extract by successively partitioning with different solvents. Mice were induced diabetes type 2 by streptozotocin (STZ) at a low dose and treated with EtOAc fraction of M. pudica leaves at 50 mg/kg and 100 mg/kg b.w for 60 days. After 24 hours of the final dose of therapy, the mice were sacrificed to extract blood and kidney tissues for biochemical and histopathological analysis. Results: The EtOAc fraction of M. pudica leaves showed strong activity in improving glucose concentration in the oral glucose tolerance test. Our results showed that EtOAc fraction significantly decreased levels of glucose, total cholesterol (TC), low-density lipoprotein (LDL), and triglyceride (TG) and increased the level of high-density lipoprotein (HDL), and protected kidneys against damage in mice. EtOAc fraction also increased the levels of antioxidant enzymes, including catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD), and decreased malondialdehyde (MDA) formation and pro-inflammatory cytokines (IL-1β and TNF-α) in kidney tissues. Moreover, the renoprotective effect was also observed in the histopathological analysis. Conclusion: Our findings support the fact that the EtOAc fraction of M. pudica leaves has potent anti-diabetic nephropathy activity by decreasing pro-inflammatory cytokines and improving antioxidant levels.

Bioassay-Guided Characterization, Antioxidant, Anti-Melanogenic and Anti-Photoaging Activities of Pueraria thunbergiana L. Leaf Extracts in Human Epidermal Keratinocytes (HaCaT) Cells

Although the roots and flowers of P. thunbergiana are known to have various physiologically active effects, studies on the anti-melanin production and anti-photoaging effects of its leaf extracts and cellular mechanisms are still lacking. In this study, we evaluated the possibility of using Pueraria thunbergiana leaves as a natural material for skin whitening and anti-aging-related functional cosmetics. The 30% ethyl alcohol (EtOH) extract from P. thunbergiana leaves was fractionated using n-hexane, ethyl acetate (EtOAc), butanol, and aqueous solution to measure their whitening, and anti-aging effects. The EtOAc fraction contained a high content of phenolic and flavonoids and showed higher 1,1-diphenyl-2-picryhydrazyl (DPPH) and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activities than the other fractions. It was also confirmed that the EtOAc fraction markedly inhibited α-melanocyte stimulating hormone (α-MSH)-induced melanogenesis in B16F10 melanoma cells. In addition, the EtOAc fraction showed a protective effect against ultraviolet B (UVB) in HaCaT cells and increased the collagen synthesis that was decreased due to UVB exposure. Matrix metalloproteinase-1 (MMP-1) activity and MMP-1 protein expression were reduced in human epidermal keratinocytes (HaCaT) cells. These results indicate that the EtOAc fraction has superior antioxidant activity, anti-melanogenesis, and anti-photoaging effects compared to the other fractions. Therefore, in this study, we confirmed the potential of P. thunbergiana leaf extract as a functional cosmetic ingredient, and it can be used as basic data for the physiological activity of P. thunbergiana leaf extracts.

A preliminary study of the chemical composition and bioactivity of Bombax ceiba L. flower and its potential mechanism in treating type 2 diabetes mellitus using ultra-performance liquid chromatography quadrupole-time-flight mass spectrometry and network pharmacology analysis.

This study aimed to preliminary investigate the phytochemistry, bioactivity, hypoglycemic potential, and mechanism of action of Bombax ceiba L. flower (BCF), a wild edible and food plant in China. By using methanol extraction and liquid-liquid extraction, the crude extract (CE) of BCF and its petroleum ether (PE), dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol (n-BuOH), and aqueous (AQ) fractions were obtained, and their chemical components and biological activities were evaluated. Further high-performance liquid chromatography (HPLC) analysis was carried out to identify and quantify the active constituents of BFC and its five fractions, and the phytochemical composition of the best-performing fraction was then analyzed by ultra-performance liquid chromatography quadrupole-time-flight mass spectrometry (UPLC/Q-TOF-MS). Finally, a network pharmacology strategy based on the chemical profile of this fraction was applied to speculate its main hypoglycemic mechanism. Results revealed the excellent biological activities of BCF, especially the EtOAc fraction. In addition to the highest total flavonoid content (TFC) (367.72 μg RE/mg E) and total phenolics content (TPC) (47.97 μg GAE/mg E), EtOAc showed the strongest DPPH⋅ scavenging ability (IC50 value = 29.56 μg/mL), ABTS⋅+ scavenging ability (IC50 value = 84.60 μg/mL), and ferric reducing antioxidant power (FRAP) (889.62 μg FeSO4/mg E), which were stronger than the positive control BHT. EtOAc also exhibited the second-best α-glucosidase inhibitory capacity and second-best acetylcholinesterase (AChE) inhibitory capacity with the IC50 values of 2.85 and 3.27 mg/mL, respectively. Also, EtOAc inhibited HepG2, MCF-7, Raw264.7, and A549 cell with IC50 values of 1.08, 1.62, 0.77, and 0.87 mg/mL, which were the second or third strongest in all fractions. Additionally, HPLC analysis revealed significant differences in the compounds’ abundance between different fractions. Among them, EtOAc had the most detected compounds and the highest content. According to the results of UPLC/Q-TOF-MS, 38 compounds were identified in EtOAc, including 24 phenolic acids and 6 flavonoids. Network pharmacological analysis further confirmed 41 potential targets of EtOAc in the treatment of type 2 diabetes, and intracellular receptor signaling pathways, unsaturated fatty acid, and DNA transcription pathways were the most possible mechanisms. These findings suggested that BCF was worthwhile to be developed as an antioxidant and anti-diabetic food/drug.

Phytoconstituents and Antimycobacterial Activities of Root Extracts and Fractions from Vernonia glabra, (Steetz) Vatke

Vernonia glabra is used by Malawian traditional healers to treat a myriad of ailments including shingles, diabetes, dysentery, diarrhoea, cough, wounds, gonorrhoea, and gastrointestinal problems. This study evaluated phytoconstituents and the in vitro antimycobacterial activities of the crude extracts and fractions from roots of Vernonia glabra. Agar disk diffusion and modified broth microdilution assays were used to determine the antimycobacterial activities of dichloromethane (DCM) and ethyl acetate (EtOAc) crude extracts and fractions from the roots. Gas chromatography-mass spectrometry (GC-MS) and Fourier-transform infrared spectroscopy (FTIR) analyses were used to characterise the major compounds in the crude extracts and fractions. The EtOAc fraction exhibited the highest antimycobacterial activity against Mycobacterium tuberculosis H37Rv. The EtOAc fraction achieved a minimum inhibitory concentration (MIC) of 4.88 μg/mL and a half-maximal inhibitory concentration (IC50) value of 38.37 μg/mL, whereas gentamycin, a standard drug, achieved MIC and IC50 values of 3.13 μg/mL and 3.88 μg/mL, respectively. GC-MS and FTIR analyses of the EtOAc fractions revealed the presence of hexadecenoic acid, methyl ester (0.39%), diisooctyl phthalate (24.56%), oxalic acid, allyl undecyl ester (4.66%), bis(2-ethylhexylphthalate) (5.41%), octacosane (51.12%), tetradecane (12.63%), phenol, 3,5-bis(1,1-dimethylethyl) (0.07%), and 2-hydroxy-4-methylbenzaldehyde (6.47%). The identified compounds are reported in the literature to possess antimicrobial and anticancer activities. It was evident from the comparison of the hill slope (HS) values of the dose-response curves of the fraction (−1.005) and its crude extract (−1.322) that repeated fractionation improved the activity. Therefore, this study showed that the roots of Vernonia glabra present a potential source of new leads for antimycobacterial drugs.

Metabolomic profile and in vitro evaluation of the cytotoxic activity of Asphodelus microcarpus against human malignant melanoma cells A375

Melanoma is a huge worldwide health problem that must be handled more effectively with better therapeutic options. As a result, new treatment drugs are required to treat this condition. The goal of this study was to investigate the cytotoxic activity of the anthraquinone-rich fractions obtained from Asphodelus microcarpus against human melanoma cell A375. On these melanoma cell lines; the cytotoxicity of these fractions had never been studied before. Liquid chromatography linked to mass spectrometry (LC-MS-MS) and Nuclear Magnetic Resonance was used to determine the chemical profiles of these fractions. The cytotoxicity of the fractions studied was determined by measuring cell viability and calculating IC 50 values. Both ethyl acetate (EtOAC) and the precipitate fractions (PPT) exhibited selective cytotoxicity on human melanoma A 375 cell line with IC 50 values of 83 and 65 µg/mL, respectively. The antiproliferative properties of EtOAc fraction and PPT were supported by a noticeable decrease in cell numbers during the G2/M cell cycle arrest. Our findings suggest that the anthraquinone content of A. microcarpus tubers is responsible for its anti-proliferative and apoptotic properties and that further in vivo investigations should be conducted to establish the viability of using them to treat human melanomas.

대용차 개발을 위한 착색된 배(Pyrus serotina)열매로부터 항산화 화합물의 분리 동정

For alternative tea development, 2 kg of dried stained Pyrus serotina was extracted with 70 % MeOH aqueous solution, and the extract was solvent-fractionated with EtOAc, n-BuOH and Water. Two types of bioactive compounds were separated from the EtOAc fraction, by purification with silica gel and octadecyle silica gel (ODS) column chromatography. The chemical structure of the compounds were identified as quercetin (1) and geniposide (2) by analyzing spectral data with NMR, MS and IR. For the evaluation of antioxidant activity, DPPH inhibitory efficacy test and ROS production inhibitory efficacy test were performed. First, as a result of measuring the DPPH inhibitory efficacy of two samples up to the highest concentration of 50 ppm in the DPPH inhibition efficacy test, the IC50 values of NICS-1 and NICS-2 were measured to be 25.554 ppm and 22.943 ppm, respectively. In ROS production inhibitory efficacy test, the ROS production was reduced in a concentration-dependent manner in both samples, and statistically significantly inhibited ROS production compared to the UV irradiation group. According to recent reports, this study is expected to develop eco-friendly sustainable health drinks using pears and cheese and functional raw materials using pears and teeth through organic cosmetics.