Dichloromethane Fraction Research Articles

Application of microemulsion in extract of the stalk of the Manilkara zapota (L.) P. Van Royen plant and evaluation of antioxidant activity

The aim of this work was to evaluate the antioxidant activity of the stem of the Manilkara zapota plant and its application in a microemulsion. To this end, the crude extract was obtained using the percolation method in 70% (v/v) ethanol and then, by means of liquid-liquid partitioning, three distinct fractions were obtained: dichloromethane fraction, ethyl acetate fraction and aqueous fraction, respectively. Their antioxidant activity was analyzed using the ABTS radical scavenging method and total phenolic compounds using the Folin-Ciocalteau method. The microemulsions were obtained following the formulation described by MALAKAR (2011). The fraction with the most pronounced antioxidant activity and the highest concentration of total phenolics and the crude extract were used for application in microemulsions. The best results for antioxidant activity were 22.22±0.20 and 43.09±1.07 (103 μM Trolox/g) for the crude extract and ethyl acetate fraction, respectively. The total phenolic content was 109.90±7.80 and 66.57±3.51 (mg EAG/g) for the crude extract and ethyl acetate fraction, respectively. After application in microemulsion, better antioxidant activity were observed for the ethyl acetate fraction (176.88±3.74 x103 μM Trolox/g) and crude extract (112,40±2,27 x103 μM Trolox/g) and higher concentrations of total phenolics for the ethyl acetate fraction (184.55±9.74 mg EAG/g). In this way, it was possible to confirm the antioxidant activity of the stem of the Manilkara zapota plant, as well as to potentiate it through fractionation and application in a microemulsion system.

Chemical Constituents, Hypolipidemic, and Hypoglycemic Activities of Edgeworthia gardneri Flowers.

The flowers of Edgeworthia gardneri are used as herbal tea and medicine to treat various metabolic diseases including hyperglycemia, hypertension, and hyperlipidemia. This paper investigate the chemical constituents and biological activities of ethanolic extract and its different fractions from E. gardneri flowers. Firstly, the E. gardneri flowers was extracted by ethanol-aqueous solution to obtain crude extract (CE), which was subsequently fractionated by different polar organic solution to yield precipitated crystal (PC), dichloromethane (DCF), ethyl acetate (EAF), n-butanol (n-BuF), and residue water (RWF) fractions. UHPLC-ESI-HRMS/MS analysis resulted in the identification of 25 compounds, and the main compounds were flavonoids and coumarins. The precipitated crystal fraction showed the highest phenolic and flavonoid contents with 344.4 ± 3.38mg GAE/g extract and 305.86 ± 0.87mg RE/g extract. The EAF had the strongest antioxidant capacity and inhibitory effect on α-glucosidase and pancreatic lipase with IC50 values of 126.459 ± 7.82 and 23.16 ± 0.79 µg/mL. Besides, both PC and EAF significantly regulated the glucose and lipid metabolism disorders by increasing glucose consumption and reducing TG levels in HepG2 cells. Molecular docking results suggested that kaempferol-3-O-glucoside and tiliroside had good binding ability with enzymes, indicating that they may be potential α-glucosidase and pancreatic lipase inhibitors. Therefore, the E. gardneri flowers could be served as a bioactive agent for the regulation of metabolic disorders.

Chemical constituents from roots of Kadsura heteroclita

The dichloromethane fraction of Kadsura heteroclita roots was separated and purified by chromatographic techniques(e.g., silica gel, Sephadex LH-20, ODS, MCI column chromatography) and semi-preparative HPLC. Twenty compounds were isolated from K. heteroclita, and their structures were identified by NMR, MS, UV, and X-ray single crystal diffraction techniques. Twenty compounds were isolated from K. heteroclita, which were identified as xuetongdilactone G(1), mallomacrostin C(2), 3,4-seco(24Z)-cychmrt-4(28),24-diene-3,26-dioic acid 3-methyl ester(3), nigranoic acid(4), methyl ester schizanlactone E(5), schisandronic acid(6), heteroclic acid(7), wogonin(8),(2R,3R)-4'-O-methyldihydroquercetin(9), 15,16-bisnor-13-oxo-8(17),11E-labdadien-19-oic acid(10), stigmast-4-ene-6β-ol-3-one(11), psoralen(12),(1R,2R,4R)-trihydroxy-p-menthane(13), homovanillyl alcohol(14), 2-(4-hydroxyphenyl)-ethanol(15), coniferaldehyde(16),(E)-7-(4-hydroxy-3-methoxyphenyl)-7-methylbut-8-en-9-one(17), acetovanillone(18), vanillic acid(19) and vanillin(20). Compound 1 is a new compound named xuetongdilactone G. Compounds 2-3 and 8-20 are isolated from K. heteroclita for the first time.

Allelopathic potential of ethanolic extract and its fractions from leaves of Geonoma schottiana Mart.

Geonoma schottiana Martius (Arecaceae) is an understory palm species widely distributed in Brazil. The objective of this work was to determine the allelopathic potential of Geonoma schottiana leaf extract and its fractions of different polarities and evaluate in vitro and in vivo antioxidant activities. The germination of seeds of Lactuca sativa and Panicum maximum was tested using ethanolic leaf extract and its hexane, dichloromethane, ethyl acetate, and butanol fractions. In vitro antioxidant tests were performed through the DPPH, ABTS, FRAP, and molybdenum tests, whereas the SOD, CAT, and POX enzymatic tests were used to evaluate in vivo antioxidant activity. Chemical analyses were performed through phytochemical screening of the main groups of secondary metabolites by thin-layer chromatography and quantification of the total contents of phenolics, tannins, and flavonoids. The hexane and butanol fractions had allelopathic effects on P. maximum and low activity on L. sativa. The ethyl acetate fraction presented higher antioxidant activity in the DPPH test, whereas butanol presented higher antioxidant activity in the ABTS test. The most active fractions in terms of allelopathic effect interfered with the increases in the production of SOD and POX enzymes. The most polar fractions had higher contents of phenolic compounds. All analyzed chemical groups were found in the phytochemical screening, except the anthracene group.

Qualitative Phytochemical Screening, Fatty Acid Profile and Biological Studies of the Bark of <i>Mallotus nudiflorus</i> (Pitali) Plant

In the present study, the Mallotus nudiflorus (L.) plant has been taken to determine the in-vitro analysis to find out the therapeutic value. The bioassays of the raw extract of methanol of bark of M. nudiflorus and by Kupchan’s extraction method collecting n-Hexane (HEX), Dichloromethane (DCM), Chloroform (CHCl3), Ethylacetate (EA) and Aqueous (AQ) fractions were scrutinized to find out its therapeutic value. The findings of phytochemical screening of the methanol extract of barks revealed the presence of several secondary metabolites. By using the GC-FID method the result showed that M. nudiflorus contained four bound fatty acids and four free fatty acids. EA fraction had the maximum phenolic content among all the fractions at (133.67±0.99) mg of GAE/g where the Folin- Ciocalteu reagent was used as an oxidizing agent. The antioxidant activity was measured in terms of its ability to scavenge free radicals (DPPH assay). Among all extractives, the greatest ability to scavenge for free radicals was shown by EA extract with an IС50 value (12.08±0.15) μg/ml. In the toxicity of brine shrimp test, the HEX fraction had the maximum toxicity with an LC50 value of (0.12±0.01) μg/ml. Cell cytotoxicity was observed for sample CHCl3 and EA on both the Vero (kidney epithelial cells taken from an African green monkey) and HeLa (a human cervical cancer cell) cell line. All the fractions were subjected to in vitro microbial screening, which revealed that DCM, CHCl3, and EA fractions showed growth inhibition, particularly against various Gram-positive and Gram-negative bacteria by disc diffusion method. The maximum zone of inhibition in the antimicrobial activity was produced by CHCl3 fraction against Staphylococcus aureus (17 mm). The results of in vitro experiments have demonstrated that the extracts from the barks of M. nudiflorus have great potential for medicinal uses and might be studied for further chemical exploration.

Solanum aculeastrum Dunal berries: Phytochemical profiling and GC-MS analysis of methanolic extract and n-hexane, dichloromethane, ethyl acetate and n-butanol fractions

Background: Solanum aculeastrum is reportedly used in several diseases including gonorrhea, bronchitis, jigger infestations and wounds, and cancers. We conducted an exhaustive phytochemical and GC-MS profiling of its methanol extract and solvents’ fractions. Methods: About 4500g of dried berries of S. aculeastrum was extracted with methanol, part of which was partitioned into fractions of n-hexane, dichloromethane, ethyl acetate, n-butanol and aqueous fractions. These fractional preparations were subjected to phytochemical and GC-MS profiling except aqueous fraction. Results: The percentage yield of 430.69g of methanol extract of berries of S.aculeastrum was 9.57%w/w, while the yields of dried 19.4g of n-hexane, 38.4g of DCM, 6.4g of ethyl acetate, 81.03g of n-butanol and 21.2g of the freeze-dried aqueous fractions were 7.08%w/w, 14.03 %w/w, 2.33%w/w, 29.60%w/w and 7.74%w/w respectively. The relative presence of glycosides, alkaloids, tannins, flavonoids, terpenoids, phenols, saponins were confirmed except quinones. GC-MS profiling of methanol extract identified 32 compounds including alkane, alcohol, carbohydrate, fatty acids, terpenoids, glycerides, vitamins and some unclassified compounds. 25 compounds including terpenoids, alkene, fatty acids, vitamin and unclassified compounds were identified in n-hexane fractions. The DCM fraction yielded 20 compounds including isoprenoid, terpenoids, amino acids, carboxylic acid and unclassified compounds. 22 compounds were identified in ethyl acetate fraction including phenol, fatty acids, alcohol, terpenoids, glycerolipid and unclassified compounds. The n-butanol fraction yielded 11 compounds including fluorinated aromatic substance, hormonal antineoplastic and a non-steroidal anti-inflammatory agent. Conclusion: This study has further elaborated on the bioactive compounds in berries of S. aculeastrum, to aid robust understanding of its pharmacologic activities.

Evaluation of ethanol extract of Allium cepa and fractions on haematological parameters in doxorubicin-induced Cardiotoxicity using Wistar rats

This research study evaluated the effects of Ethanol extract of Allium cepa (A. cepa) popularly known as Onions and its fractions on haematological parameters in doxorubicin-induced cardiotoxicity using Wistar rats. A total of 45 Wistar rats weighing between 150-200 g were recruited for this study. They were randomly divided into 9 groups of 5 animals each; as follows: Group 1: served as control and received 10 ml /kg/ body weight (b.w) per oral (p.o) of normal saline. Group II: received 3.5 mg / kg b.w intraperitoneal (i.p) of Doxorubicin (Dox). Group III: received 500 mg/kg/ b.w p.o of vitamin E plus Dox. Group IV: received 1000 mg/kg/ b.w of crude extract (C.E) of A. cepa plus Dox. Group V: received 1000 mg/kg b.w of n-Hexane fraction of A. cepa plus Dox. Group VI: received 1000 mg/kg b.w of dichloromethane (DCM) fraction of A. cepa plus Dox. Group VII received 1000 mg/kg b.w of ethyl acetate (E.A) fraction of A. cepa plus Dox. Group VIII: received 1000 mg/kg b.w of methanol (METH) fraction, and Group IX received Combinations of C.E of A. cepa + vitamin E and Dox respectively for 14days. Groups I and II treatments lasted for 14 days, while treatments for groups III-IX lasted for 16 days (making 14 days for respective treatments and additional 2 days for doxorubicin administered once 48 hourly before sacrificing. All substances in this study were administered orally except doxorubicin that was administered intraperitoneally. Haematology results obtained from this study showed Dox administration negatively altered full blood count; Red blood cell (RBC), White blood cell (WBC) and Platelets levels significantly (p<0.05). However, pre-treatment administration with Allium cepa showed significant (p<0.05) improvement of full blood count levels from the effects of singular administration of Dox. Injury to the myocardium was also observed on histological examination in the Dox treated group. Our findings indicate that haematological parameters RBC, WBC, Platelets could be useful diagnostic tools in prediction and evaluation of Cardiotoxicity. Haematology results obtained from this study showed Doxorubicin administration significantly altered (p<0.05) full blood count; Red blood cell (RBC), White blood cell (WBC) and Platelets levels. However, pre-treatment administration with Allium cepa showed significant (p<0.05) improvement of full blood count levels from the effects of singular administration of Doxorubicin. Injury to the myocardium was also observed on histological examination in the Doxorubicin treated group. Our findings indicate that haematological parameters RBC, WBC, Platelets could be useful diagnostic tools in prediction and evaluation of Cardiotoxicity.

Cytotoxic and antiviral activities of Jatropha variegata and Jatropha spinosa in relation to their metabolite profile

Jatropha variegata and Jatropha spinosa (family: Euphorbiaceae) are utilized in Yemeni traditional medicine to treat respiratory tract infection and in different skin conditions such as wound healing, as antibacterial and hemostatic. In this study, we evaluated the cytotoxicity and the antiviral activities of the methanolic J. variegata (leaves: Ext-1, stems: Ext-2, and roots: Ext-3), and J. spinosa extracts (aerial parts: Ext-4 and roots: Ext-5), in addition to their methylene chloride fractions of roots extracts (F-6 and F-7, respectively). All samples were tested against three human cancer cell lines in vitro (MCF-7, HepG2, and A549) and two viruses (HSV-2 and H1N1). Both plants showed significant cytotoxicity, among them, the methylene chloride fractions of roots of J. variegata (F-6) and J. spinosa roots (F-7) showed the highest activity on MCF-7 (IC50 = 1.4 and 1 μg/mL), HepG2 (IC50 = 0.64 and 0.24 μg/mL), and A549 (IC50 = 0.7 and 0.5 μg/mL), respectively, whereas the IC50 values of the standard doxorubicin were (3.83, 4.73, and 4.57 μg/mL) against MCF-7, HepG2, and A549, respectively. These results revealed that the roots of both plants are potential targets for cytotoxic activities. The in vitro results revealed potential antiviral activity for each of Ext-3, Ext-5, F-6, and F-7 against HVS-2 with IC50 of 101.23, 68.83, 4.88, 3.24 μg/mL and against H1N1 with IC50 of 51.29, 27.92, 4.24, and 3.06 μg/mL respectively, whereas the IC50 value of the standard acyclovir against HVS-2 was 83.19 μg/mL and IC50 value of the standard ribavirin against H1N1 was 52.40 μg/mL .The methanol extracts of the roots (Ext-3 and Ext-5) of both plants were characterized using UPLC/MS. A total of 73 metabolites were annotated, including fourteen diterpenoids, eleven flavonoids, ten phenolic acid conjugates, twelve fatty acids and their conjugates, five triterpenes and steroids, two sesquiterpenes, and six coumarins. The cytotoxicity and antiviral activities determined in the present work are explained by the existence of flavonoids, coumarins and diterpenes with commonly known cytotoxicity and antiviral activities.

HPLC-based cytotoxicity profiling and LC-ESIQTOF-MS/MS analysis of Helichrysum leucocephalum

IntroductionHelichrysum leucocephalum Boiss. (Asteraceae) is an endemic plant to Iran. No reports have studied the cytotoxicity of the plant. The current study aimed to evaluate the cytotoxicity of H. leucocephalum collected from Fars province (Iran) against MCF-7 and HDF cell lines using HPLC-based activity profiling and to annotate the active constituents by LC-ESIQTOF-MS/MS. MethodsH. leucocephalum was collected from three locations in Fars province. The dried flowers and leaves were separately extracted by percolation using methanol. The crude extracts were fractionated by liquid-liquid partitioning with dichloromethane (DCM) and aqueous methanol. The cytotoxicity of the fractions was evaluated against MCF-7 and HDF cells by Alamarblue assay. HPLC-based activity profiling was used to track the active constituents. LC-MS dereplication strategy was used for the annotation of the compounds in the active time window. LC-MS data were preprocessed by MZmine 3.3.0 and submitted to multivariate analysis to compare the differences and similarities in the metabolites of the samples. ResultsThe DCM fractions showed a dose-dependent cytotoxicity against the cancerous cells (IC50s, 9.8–105.1 μg/ml). In general, the metabolites of the flowers and their cytotoxicity were higher than the leaves. LCESIMS/MS analyses revealed that prenylated and geranylated α,β-unsaturated spiroketal phloroglucinols were among the active constituents. ConclusionIt can be concluded that H. leucocephalum is a rich source of phloroglucinol derivatives with cytotoxic activities. Further phytochemical analysis is needed to characterize the bioactive components.

Characterisation of bioactive compounds from leaves of Smilax fluminensis Steud. by gc-MS and LC-DAD-MS, and analysis of its antioxidant and ecotoxicological activities

Smilax fluminensis Steud. is distributed in Central and South America, commonly named as ‘salsaparilha’ or ‘japicanga’. In the present work, the chemical composition was determined, antioxidant and cytotoxic effects were evaluated for the ethanol extract (EE) and fractions from leaves. Fatty acid esters and phytol were characterised in the hexane (HEX) fraction. O-glycosylated flavonols and flavones, caffeic acid derivatives, and steroidal saponins were annotated for EE, and dichloromethane (DCM), ethyl acetate (AC), and hydroethanol (HE) fractions. The samples contain flavonoids and phenolic compounds, and the AC fraction displayed the biggest concentration of these substances. Antioxidant potential was observed in all samples, being especially pronounced in the AC fraction by DPPH and FRAP assays, with IC50 values of 8.18 and 2.35 µg/mL, respectively. AC and HEX fractions showed 35% and 5% lethality at 1000 μg/mL, in the Artemia salina assay, and the other samples did not show a toxic effect.

Chymotrypsin and Trypsin Inhibitory Activity of Some Medicinal Plants Collected from Rize (Türkiye).

In this research, the evaluation of in vitro chymotrypsin and trypsin inhibitory activities of ten plant species collected from Rize were aimed, and fractions that showed strong activity were analyzed through HPLC. Daphne pontica L. and Mentha longifolia (L.) L. were found to have the highest chymotrypsin inhibitory activities (87.75 and 84.24 % inhibition). Similarly, the highest trypsin inhibitory activity was observed in D. pontica (%99.93 inhibition), followed by Sambucus ebulus L. flowers (87.47 % inhibition). Extracts showing strong enzyme inhibition were fractioned and subjected to activity tests. The highest chymotrypsin inhibitory activity was observed in the n-hexane fraction of D. pontica (%80.70 inhibition), while the highest trypsin inhibitory activity was found in the n-butanol fraction of S. ebulus (%86.81 inhibition). HPLC studies determined that the 80 % ethanol extract of D. pontica and its dichloromethane and ethyl acetate fractions contained umbelliferone. It was found that chlorogenic acid was present in the 80 % ethanol extracts of S. ebulus flowers. M. longifolia was found to contain chlorogenic acid, caffeic acid, luteolin-7-glucoside, and rosmarinic acid. M. longifolia has been identified as the plant exhibiting the highest antioxidant activity in ABTS and CUPRAC tests, consistent with its high phenolic and flavonoid content.

ANTIOXIDANT ACTIVITY OF THE ACTIVE FRACTION OF MANGOSTEEN RIND EXTRACT (GARCINIA MANGOSTANA)

Objective: Determination of the IC50 value of the active Fraction was carried out to see the antioxidant activity of the ethanol extract of mangosteen peel (Garcinia mangosteen) Mangosteen peel is known to have very strong antioxidant activity. The research began with maceration of mangosteen peel with ethanol solvent. Then, the mangosteen peel extract was fractionated with three solvents, namely those with different polarities, namely water, dichloromethane, and n-hexane.
 Methods: The ethanol extract of mangosteen rind was further fractionated using n-hexane, dichloromethane, and water, then the solvent was removed by evaporation. The three resulting fractions were measured for antioxidant activity using the DPPH method with Quercetin as a comparison. The active Fraction with the highest IC50 value is then compared with the extract, and the comparison is seen with TLC. Next, the Fraction was tested with GC to see the remaining solvent. And then continued with determining alpha mangostin levels in all fractions using UHPLC.
 Results: IC50 value for each n-hexane fraction was 50.65µg/ml, the dichloromethane fraction was 34.66µg/ml, and the water fraction was 45.72µg/ml. The results of the solvent test showed that there was no residual solvent in the dichlormethane fraction, as seen from the chromatogram results. The results of the assay showed the following results: n-Hexan fraction 25.18%, DCM fraction 31.23%.
 Conclusion: The dichloromethane fraction showed the highest antioxidant activity with the best IC 50 value and had higher levels of alpha mangostin than the water and n-hexane fractions.

Metabolic Profiling, GC-MS, LC-ESI-MS/MS Analysis, Phenolics Isolation and Biological Evaluation of the Aerial Parts Extracts of Felicia abyssinica L.

Felicia abyssinica L., family Asteraceae, is widely used in folk medicine. This represents the first study to investigate its phytoconstituents and pharmacological effects. Phytoconstituents identified by GC-MS, LC-ESI-MS/MS-based metabolomics, and NMR (1D & 2D). GC-MS of the (FAMEs) revealed mainly the identification of 55 fatty acids. LC-ESI-MS/MS analysis resulted in the tentative identity of 13 compounds representing flavonoids, phenolics, and fatty acids. Ethyl acetate fraction exhibited the highest total flavonoids 66.19 mg/mL Rutin equivalent, while the methanolic fraction showed the highest phenolics 87.70 mg/mL gallic acid equivalent, and the total condensed tannins were 64.35 μg CE/mg catechins equivalent. A flavonoid and a cinnamic acid derivative were identified as quercetin 3-O-(2'''-O-acetyl) rutinoside (Mumikotin A) (1) and Methyl sinapate (2). Biological evaluation of antioxidant and cytotoxic activities was carried out. Cytotoxicity was examined on HepG-2 cell lines where the average cell viability was 91.42 % and 52.48 % for concentrations 10 and 100 μg/mL respectively. Methylene chloride and methanolic fractions showed the highest antioxidant activity 225 μg/mL Ascorbic acid equivalents. It is hypothesized that high phenolics, flavonoid content, and oxygenated identified compounds contribute to the antioxidant activity and can be regarded as a promising species for nutraceuticals active antioxidants with potential value for remedy.

Phytochemical Contents and Anti-Colon Cancer Activities of Macadamia integrifolia Maiden & Betche Leaves and Husks Extracts

The macadamia (Macadamia integrifolia Maiden & Betche, Proteaceae Family) leaves and husks were the by-products from macadamia nut production which have few reports on the phytochemicals and pharmacological activities. This study aimed to investigate the possibility of using these by-products in drug discovery by examining their phytochemicals contents, as well as their anti-colon cancer activities. Dry macadamia leaves and husks were extracted and fractionated to obtain the crude extracts (EtOH), dichloromethane (DCM), ethyl acetate (EtOAc) and water fractions. Colorimetric methods were used to determine the total amounts of phytochemicals. The anti-colon cancer activities of the extracts against human colorectal carcinoma (HCT116) cell line were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results showed that each gram of the samples contained total phenolics between 2.9-12.9 mg equivalent to gallic acid, total flavonoids 30.1-89.7 mg equivalent to rutin, and total triterpenoids 228.4-640.9 mg equivalent to ursolic acid. The DCM and EtOAc fractions of the husks exhibited the highest anti-colon cancer activity, with half maximal inhibitory concentrations (IC50) of 39.1 ± 4.3 and 56.9 ± 15.0 µg/mL, respectively. The EtOAc and water fractions of the leaves had IC50 values of 130.9 ± 17.4 and 127.8 ± 7.9 µg/mL, respectively. The IC50 values of other samples were greater than 200 µg/mL, while the IC50 of sorafenib, a positive control, was 5.95 ± 1.26 µg/mL. In conclusion, the leaves and husks of macadamia contained high amounts of several phytochemicals and exhibited anti-colon cancer activity against HCT116. These findings suggested that the extracts from these wastes may have potential in the discovery of anticancer agent research. Keywords: Macadamia integrifolia, Leaf, Husk, Colon cancer, Phytochemical

Antioxidant and cytotoxic properties of Combretum paniculatum and Fadogia agrestis extracts on prostate and cervical cancers cell lines

The medicinal plants used in Burkina Faso are essential source of research for bioactive compounds against prostate and cervical cancer, which are major health problems in Burkina Faso and worldwide. This was done by hydro-acetic maceration of the powder of the leafy branches of Combretum paniculatum (C. paniculatum) and Fadogia agrestis (F. agrestis). These extracts were then fractionated using petroleum ether, dichloromethane, ethyl acetate, and n-butanol. A phytochemical screening of these extracts was performed using High-Performance Thin Layer Chromatography (HPTLC). Antioxidant activity was evaluated using 2,2-Diphenyl-1-picrylhydrazyl (DPPH∎) and 2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic Acid (ABTS∎+). The extracts cytotoxicity was assessed by the MTT (Mitochondrial Tetrazolium Test) method on the LNCaP cell lines of prostate cancer and HeLa of cervical cancer. Phytochemical analysis of extracts and fractions revealed the presence of tannins, flavonoids, and steroids in these plants. For antioxidant activity, the ethyl acetate fraction of F. agrestis and the n-butanol fraction of C. paniculatum gave the best inhibition of DPPH∎ with IC50 values of 0.246 ± 0.045 and 0.097 ± 0.008 μg extract/mL, respectively. Using the ABTS∎+ method, the ethyl acetate fractions of C. paniculatum (1.1009 ± 0.0181 μM ET/g) and F. agrestis (0.0316 ± 0.0007 μM ET/g) showed the best activity. In addition, the fractions of dichloromethane (DCM) of F. agrestis (IC50 = 24.69 ± 9.51 μg/ml) and ethyl acetate (AE) of F. agrestis (IC50 = 129.9 ± 6.49 μg/ml) had better activity, respectively on the prostate and cervical cancer LNCaP lines. These properties could justify the use of these plants in traditional medicine.

Phytochemical composition of aerial parts and roots of Pfaffia glomerata (Spreng.) Pedersen and anticholinesterase, antioxidant, and antiglycation activities.

The Pfaffia glomerata, a plant popularly called Brazilian ginseng, is widely used in Brazil for the treatment of various pathologies, including those associated with the Central Nervous System. 20-hydroxyecdysone (20E), a phytosteroid present in this plant, can promote adaptogenic effects in the organism, providing greater body resistance to stressors. This study aimed to evaluate the phytochemical composition and the anticholinesterase, antioxidant, and antiglycation effects of extracts and fractions of aerial parts and roots of P. glomerata, also analyzing their possible cytotoxic effects. The fractions were obtained by partitioning methanol extracts from the aerial part and roots of P. glomerata with hexane, dichloromethane, ethyl acetate, n-butanol, and water. The samples were initially tested in anticholinesterase, antioxidant, and antiglycation assays, and the most promising samples were submitted for cytotoxicity and chromatographic analyses. Mass spectrometry and chromatography methods revealed that 20E was the main compound in the dichloromethane fractions, there being 35% more 20E in the aerial part (APD) than in the roots (RD). Added to the higher concentration of 20E, the APD fraction also presented more promising results than the RD fraction in anticholinesterase and antioxidant analyses, indicating that their effects may be related to the concentration of 20E. These same fractions showed no hemolytic effects but were cytotoxic in high concentrations. These new findings contribute to scientific information about P. glomerata and open more perspectives for the understanding of its therapeutic properties, allowing the association of biological activity with the presence of 20E.

Investigating bioactive phytochemicals in bulb and shoot of Allium longisepalum Bertol. from Iran

This study investigated a wide range of bioactive compounds in bulb and shoot of Allium longisepalum Bertol. from Iran. The organs were first extracted with methanol, followed by hexane, dichloromethane, chloroform, ethyl acetate, and butanol, and phytochemicals in fractions were analysed using GC/MS. Butanol exhibited the highest yield percentages in both organs. Palmitic and oleic acid were found in all fractions, while heptadecene-(8)-carboxylic acid, stearic acid, and myristic acid were detected in at least three fractions. Each fraction contained exclusively unique metabolites found only in itself. Hexane, dichloromethane, and chloroform fractions contained specific metabolites with potential antioxidant, allelopathic, antibacterial, and anti-inflammatory effects. Ethyl acetate and butanol fractions revealed significant compounds that have potential in terms of their anticarcinogenic, anti-inflammatory, hair growth-enhancing, and antimicrobial properties, in addition to containing compounds not previously identified in plants. The study analyzes bioactive compound profiles in A. longisepalum, highlighting its chemical diversity and therapeutic potential.

Chemical Analysis of SU-Eohyeol Pharmacopuncture and Its In Vitro Biological Activities.

Introduction: Pharmacopuncture (PA) is widely used in traditional Korean medicine to treat various diseases, including abdominal obesity, nervous system diseases, and musculoskeletal disorders. In the present study, we attempted to identify the chemical components of SU-Eohyeol PA (SUEHP), comprising extracts of eight medicinal herbs and Cervi Parvum Cornu, using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) and evaluated the in vitro anti-inflammatory and antioxidant activities of SUEHP. Methods: Volatile components of SUEHP were identified by GC-MS analysis of the n-hexane, dichloromethane (DCM), and distilled water-acetonitrile (DW-CAN) solvent fractions. LC-MS was performed to identify small metabolites of SUEHP using a water-methanol solvent fraction. In vitro anti-inflammatory and antioxidant potential of SUEHP was evaluated using cell-free biochemical assays and a BV2 microglial cell culture system. Results: GC-MS of SUEHP using the n-hexane, DCM, and DW-ACN solvent fractions detected 32 components. The major components (>3%) were 9-octadecenamide (25.9%) and eicosane (5.4%) in n-hexane fraction, 9-octadecenamide (29.1%) and cis-11-octadecenoic acid (3.1%) in DCM fraction, and 9-octadecenamide (31.1%) and 9-octadecenoic acid (17.9%) in DW-CAN fraction. LC-MS of SUEHP using a water-methanol solvent fraction detected 36 primary metabolites, including (a)symmetric dimethylarginine, L-pyroglutamic acid, n,n-dimethylglycine, n-acetyl-L-aspartic acid, and amino acids. Enrichment analysis and subsequent network analysis of the primary metabolites suggested their association with neurodegenerative diseases, including Alzheimer's disease and schizophrenia. Cell-free biochemical assays and molecular signaling studies of lipopolysaccharide-stimulated BV2 murine microglial cells demonstrated the anti-inflammatory and antioxidant activities of SUEHP. Conclusion: The present study identified the biochemically active components of SUEHP and suggested their therapeutic potential against diseases related to inflammatory and oxidative stress.

Chromatographical fingerprint analysis, in silico investigation, and identification of hepatoprotective active compounds from Capparis decidua Edgwe (Forssk.)

Liver disorders are globally distributed with a high burden of disease costs. The methanolic extract of Capparis decidua stem part and its dichloromethane (DCM) and ethylacetate fractions showed significant hepatoprotective activity, so this study aimed to develop gas chromatography/mass spectrometry (GC/MS), high-performance thin layer chromatography (HPTLC) fingerprint profile and in silico docking of identified compounds from both fractions responsible for hepatoprotective activity. Hexadecanoic acid, 9-octadecenoic acid, 9,12,15-octadecatrienoic acid, and dihydrolinalool are known hepatoprotective and antioxidant agents that were identified from both fractions by GC/MS, they have various degrees of hepatoprotective and antioxidant properties, this is the first time to identify hexadecanoic acid in C. decidua stem. Both fractions were chromatogramed with known hepatoprotective standard compounds, they were rutin, quercetin, kaempferol, ferulic acid, oleanolic acid, and β-sitosterol; β-sitosterol and oleanolic were isolated and identified in the DCM fraction by HPTLC, but nothing of these standard compounds were identified in the ethylacetate fraction. According to International Council for Harmonization guidelines, the HPTLC fingerprint profile method was validated to confirm our isolation and identification process of β-sitosterol and oleanolic acid in the DCM fraction, also their contents were quantified and they were 9.21 and 7.73 μg/mg, respectively, in the total dry fraction weight, this was the first time to isolate and identify oleanolic acid from C. decidua stem part. In silico docking was carried and was found that hexadecanoic acid showed significant predicted binding properties on Nrf2, p38 mitogen-activated protein kinase, and IL-1 receptor proteins. 9-octadecenoic acid (elaidic acid) showed significant predicted interactions with IL-1 receptor, Nrf2, and peroxisome proliferator-activated receptor-α (PPAR-α) proteins. Oleanolic acid showed significant predicted bindings with p65, JNK, and NF-kb. β-sitosterol showed significant predicted interaction with IL-6α protein, while 9,12,15-octadecatrienoic acid revealed significant binding with PPAR-α protein. Therefore, we recommend considering our active compounds in the field of drug development (structure-activity relationship) and the possibility of the presence of synergism if they are combined and administered together. Moreover, we recommend liquid chromatography-mass spectrometry and nuclear magnetic resonance on both fractions to search for the possibility of the presence of other hepatoprotective compounds.

Anti-leishmanial activity of Eleutherine plicata Herb. and predictions of isoeleutherin and its analogues.

Introduction: Leishmaniasis is caused by protozoa of the genus Leishmania, classified as tegumentary and visceral. The disease treatment is still a serious problem, due to the toxic effects of available drugs, the costly treatment and reports of parasitic resistance, making the search for therapeutic alternatives urgent. This study assessed the in vitro anti-leishmanial potential of the extract, fractions, and isoeleutherin from Eleutherine plicata, as well as the in silico interactions of isoeleutherin and its analogs with Trypanothione Reductase (TR), in addition to predicting pharmacokinetic parameters. Methods: From the ethanolic extract of E. plicata (EEEp) the dichloromethane fraction (FDEp) was obtained, and isoeleutherin isolated. All samples were tested against promastigotes, and parasite viability was evaluated. Isoeleutherin analogues were selected based on similarity in databases (ZINK and eMolecules) to verify the impact on structural change. Results and Discussion: The extract and its fractions were not active against the promastigote form (IC50 > 200μg/mL), while isoeleutherin was active (IC50 = 25μg/mL). All analogues have high intestinal absorption (HIA), cell permeability was moderate in Caco2 and low to moderate in MDCK. Structural changes interfered with plasma protein binding and blood-brain barrier permeability. Regarding metabolism, all molecules appear to be CYP3A4 metabolized and inhibited 2-3 CYPs. Molecular docking and molecular dynamics assessed the interactions between the most stable configurations of isoeleutherin, analogue compound 17, and quinacrine (control drug). Molecular dynamics simulations demonstrated stability and favorable interactions with TR. In summary, fractionation contributed to antileishmanial activity and isoleutherin seems to be promising. Structural alterations did not contribute to improve pharmacokinetic aspects and analogue 17 proved to be more promising than isoeleutherin, presenting better stabilization in TR.