Crude Dichloromethane Research Articles

Monoterpene Hydroxy Lactones Isolated from Thalassiosira sp. Microalga and Their Antibacterial and Antioxidant Activities.

Two monoterpenoid lactones, loliolide (1) and epi-loliolide (2), were isolated from the crude dichloromethane extract of a microalga, Thalassiosira sp.). The structures of loliolide (1) and epi-loliolide (2) were elucidated by 1D and 2D NMR analysis, as well as a comparison of their 1H or/and 13C NMR data with those reported in the literature. In the case of loliolide (1), the absolute configurations of its stereogenic carbons were confirmed by X-ray analysis, whereas those of epi-loliolide (2) were determined by NOESY correlations. Loliolide (1) and epi-loliolide (2) were tested for their growth inhibitory activity against two Gram-positive (Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212) and two Gram-negative (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853) bacteria, as well as one clinical isolate (E. coli SA/2, an extended-spectrum β-lactamase producer-ESBL) and two environmental isolates, S. aureus 74/24, a methicillin-resistant (MRSA), and E. faecalis B3/101, a vancomycin-resistant (VRE) isolates. The results showed that none of the tested compounds exhibited antibacterial activity at the highest concentrations tested (325 μM), and both revealed low antioxidant activity, with ORAC values of 2.786 ± 0.070 and 2.520 ± 0.319 µmol TE/100 mg for loliolide (1) and epi-loliolide (2), respectively.

The Protective Effect of Juçara Fruit (Euterpe edulis Martius) Extracts on LPS-Activated J774 Macrophages.

This study investigated the anti-inflammatory effect of hydrophilic and lipophilic extracts from juçara fruits (Euterpe edulis Martius) through measurement of nitric oxide (NOx) and cytokines (IL-12p70, TNF-α, INF-γ, MCP-1, IL-6, and IL-10). J774 macrophages were stimulated with lipopolysaccharides (1µg/mL) and treated with various concentrations (1-100µg/mL) of juçara fruits extracts from crude extracts, and hexane, dichloromethane, ethyl acetate, and butanol fractions. Potential relationships between the phenolic composition of the extracts determined by LC-ESI-MS/MS and their anti-inflammatory capacity were also evaluated. Hexane and dichloromethane fractions inhibited NOx and IL-12p70 while increased IL-10. Hexane fractions also decreased IL-6 and IFN-γ production. Hexane and dichloromethane fractions showed a higher number of phenolic compounds (32 and 34, respectively) than the other extracts tested and were also the only ones that presented benzoic acid and pinocembrin. These results suggest juçara fruits compounds as potential anti-inflammatory agents, especially those of a more apolar nature.

Antinociceptive and anti-inflammatory activity of extracts and α-pyrones isolated from Cantinoa stricta.

This study evaluated the composition and the antinociceptive and anti-inflammatory activity of the crude extracts and two isolated compounds, anamarine (ANA) and 10-epi-olguine (eOL), obtained from the leaves of Cantinoa stricta (Lamiaceae). Crude ethanolic extract (EEt) and dichloromethane extract (DCM), selected based on NMR data, were submitted to pharmacological tests in male Swiss mice. The oral administration of EEt and DCM significantly reduced the second phase of formalin-induced nociception (60%), lipopolysaccharide (LPS)-induced mechanical hyperalgesia (90%), and carrageenan (Cg)-induced edema (25%). ANA and eOL, the major compounds in EEt and DCM extracts, administered orally or locally (in the paw), also reduced the LPS-induced mechanical hyperalgesia (Oral ID50 1.9 and 3.9mg/kg; Local ID50 93.4 and 677.3ng, respectively) without changing the thermal acute nociception or the motor performance of the animals. Local administration of ANA and eOL also reduced Cg-induced edema (40 and 23%, respectively). These isolated compounds did not change the mechanical hyperalgesia induced by tumor necrosis factor-α, interleukin-1β, prostaglandin E2, dibutyryl cyclic AMP, or forskolin but reversed the hyperalgesia induced by dopamine, epinephrine, and phorbol 12-myristate 13-acetate. The hyperalgesia induced by epinephrine was reversed in male but not in female mice, in which this response is not dependent on protein kinase C (PKC). These results suggest thatC. strictaextracts possess antinociceptive and anti-inflammatory activity which is related to the presence of ANA and eOL. Differently from the known analgesics, these substances seem to exert their action mainly interfering with the sympathetic component of pain, possibly with PKC.

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

Phytochemical characterization by GC-MS and in vitro evaluation of antiproliferative and antimigratory studies of Leucas aspera leaf extracts on MDA-MB-231 cell line.

Breast cancer is the most recurrently identified and one of women's prominent causes of death. Currently, researchers have turned their focus on natural chemicals from synthetic chemicals due to their environmental, economic, and health benefits. Considering this, the medicinal plant Leucas aspera was chosen for the current study. The aim of this study was to isolate and characterize secondary metabolites from L. aspera and determine the antiproliferative and antimigratory activities in the MDA-MB-231 cell line under in vitro conditions. Phytochemicals from L. aspera were isolated through sequential extraction using hexane, dichloromethane, and ethyl acetate. These extracts were qualitatively screened, subjected to FT-IR, and analyzed using GC-MS. The antiproliferative activity was determined through the MTT assay. Scratch assay was utilized to determine the antimigratory activity of the plant extracts. The phytochemical analysis revealed the presence of steroids, alkaloids, phenols, flavonoids, galactose, tannins, saponins, and amino acids in the extracts. The results of the cell viability assay indicated that the crude dichloromethane and ethyl acetate extracts inhibited cell proliferation, with inhibitory concentrations of 5 and 3 μg/ml, respectively. In contrast, the crude hexane extract did not exhibit any cytotoxicity. Furthermore, the scratch assay results showed that the plant extracts had cell migration inhibitory properties. The outcomes of the current study conclude that L. aspera possesses active therapeutic agents with strong anticancer potential, effectively impeding the proliferation and invasion of MDA-MB-231. Further studies are needed to identify the potential active agents that contribute to these activities.

Investigation of antioxidant activity of \(\textit{Zoanthus vietnamensis’}\)s crude extract and dichloromethane extract

Zoanthid is a potential natural resource with many biological activities, such as antioxidant, antiviral, anti-inflammatory, and cytotoxic activity. Therefore, this study aimed to evaluate the antioxidant activity of 70% ethanol crude extract and dichloromethane fraction from Zoanthus vietnamensis. The total polyphenol content of ethanol and dichloromethane extracts were 19.36 ± 1.37 and 24.95 ± 0.63 mg GAE/g extract, respectively. The DPPH free radical scavenging capacity increased slightly from low to high concentration. The IC50 values reached 9.974 µg/mL and 6.424 µg/mL for ethanol crude and dichloromethane extract. The potassium ferricyanide-reducing power assay result was meager, and the optical density value ranges from 0.015 to 0.105.

Nitric Oxide Inhibitory Activity of Crude Extracts and Extraction Time and Temperature Optimization of Pectin from Cissampelos pareira Leaves

This research studied crude extracts of Cissampelos pareira leaves using fi ve solvents as hexane, dichloromethane, ethyl acetate, methanol, and ethanol for nitric oxide (NO) inhibitory activity and maximize pectin yields from facile green extraction. Five crude hexane, dichloromethane, ethyl acetate, methanol and ethanol extracts were obtained from the leaves of C. pareira at 0.4%, 0.5%, 0.3%, 3%, and 1%, respectively. The hexane, dichloromethane, and ethyl acetate extracts showed the most potent decrease in NO production with half-maximal inhibitory concentration (IC50) values of 49.85±5.07, 56.26±1.49, and 64.55±2.37 (μg/mL), respectively. Concentrations of all crude extracts of C. pareira indicated no cellular toxicity. Pectin was aqueously extracted at 40 to 90 °C for 15 to 120 min and yields and quality were examined. Highest pectin yield (36.12%) was obtained after 60 min at 60 °C, with quality meeting the criteria established by the International Pectin Producers Association (IPPA) and categorized as high methoxyl pectin.

An investigation of the in vitro wound healing potential of Mitragyna speciosa (Korth.) Havil leaf ultrasound-assisted methanol crude extract and fractions

The tropical plant Mitragyna speciosa (Korth.) Havil is native to Southeast Asia, including Malaysia, Indonesia, and Thailand. Studies have identified advantageous biological attributes in the plant, such as antioxidant, anti-inflammatory, and antimicrobial, which are vital during wound healing. The current study aims to investigate the in vitro wound healing capacities of methanol crude extract and fractions of Mitragyna speciosa leaves on 3T3 fibroblast and human umbilical vein endothelial cells (HUVECs). The wound healing abilities of the extracts obtained were then assessed through cytotoxicity, scratch, and tube formation assays. The methanol crude extract and dichloromethane and ethyl acetate fractions were non-toxic against the 3T3 fibroblast cells and HUVECs at concentrations below 50 μg/mL. The scratch assay revealed that the ethyl acetate and dichloromethane fractions recorded better migration activities at low concentrations, 1.56 and 3.13 μg/mL. Furthermore, the ethyl acetate fraction recorded the highest branching length in the tube formation assay at all concentrations, thus demonstrating a strong potential for angiogenesis promotion. This study also demonstrated that targeted secondary metabolites, such as flavonoids, phenolics, and fatty acids, might possess therapeutic attributes, including anti-inflammatory, antibacterial, and antioxidant, as they are responsible for promoting blood clotting, fighting infections, and accelerating wound healing. Collectively, the results revealed that Mitragyna speciosa could be a potential candidate as a therapeutic agent for wound treatments, supporting traditional claims.

Nematicidal potential of Macrotyloma axillare 'Java' extract against Meloidogyne javanica and its metabolic profile

Plants produce chemical compounds with high nematicidal activity and potential for large-scale application in nematode control. The purpose of this study was to identify chemical compounds in crude extract of Macrotyloma axillare 'Java' and investigate their inhibitory effects on Meloidogyne javanica . Crude methanolic extract was fractionated into hexane, dichloromethane, ethyl acetate , and hydromethanol fractions. Chemical characterization of dichloromethane, ethyl acetate, and hydromethanol fractions was performed by dereplication using high-resolution mass spectrometry. The inhibitory activity of samples on the hatching of second-stage juveniles (J2) was tested at concentrations of 0%, 0.05%, 0.5%, and 5% (0, 500, 5000, 50000 ppm) and on nematode penetration and reproduction on soybean at 0.1% (1000 ppm). Distilled water was used as control in nematicidal assays. Several compounds belonging to the groups of organic acids, phenolic acids , fatty acids, flavonoids , pterocarpans, terpenoids, and sugars were identified. All fractions and concentrations reduced J2 hatching, reaching a plateau at 0.05%. Crude methanolic extract and dichloromethane and ethyl acetate fractions decreased M. javanica penetration at 10 days after inoculation, and all fractions reduced nematode penetration and female numbers at 30 days after inoculation. Extract fractions also reduced total nematode numbers, providing 49–95% control of the nematode. M. axillare 'Java' extract is rich in nematicidal compounds and has toxic effects on M. javanica , inhibiting J2 hatching and nematode penetration, development, and reproduction on soybean. • Macrotyloma axillare 'Java' is a plant that can be used to control nematodes. • 'Java' extract reduces penetration, development and reproduction of Meloidogyne javanica in soybean. • 135 compounds were identified in the 'Java' extract. • The chemical compounds of 'Java' show toxicity to Meloidogyne javanica .

Hop (Humulus lupulus L.) Specialized Metabolites: Extraction, Purification, Characterization in Different Plant Parts and In Vitro Evaluation of Anti-Oomycete Activities against Phytophthora infestans

Botanicals represent a promising source of metabolites in the search for new biofungicides. In this context, this study aimed at evaluating the in vitro anti-oomycete activity of hop (Humulus lupulus L.) extracts and metabolites against Phytophthora infestans, an oomycete causing late blight disease in Solanaceae. Crude hydro-ethanolic extracts and dichloromethane sub-extracts of different parts (cones, leaves, stems and rhizomes) were characterized by UHPLC-UV–MS and some cone specialized metabolites were purified by CPC and preparative HPLC. A commercial hop cone essential oil was also analyzed by GC–MS. All extracts succeeded in inhibiting mycelial growth and spore germination with morphological alteration of the mycelium. Extracts of leaves showed a significant anti-oomycete activity compared to the extracts of cones, stems, and rhizomes. Moreover, no difference was noticed between the crude hydro-ethanolic extract and the dichloromethane sub-extract activity, except for leaves, with the apolar sub-extract being more active than the crude one. The extracts of cones succeeded in inhibiting more P. infestans than the essential oil, which appeared to be the less active evaluated modality. Some purified prenylated phenolic compounds also inhibited P. infestans although copper sulfate, a mineral fungicide control, was still more active. This study highlights the potential use of hop by-products as biofungicides to manage P. infestans.

Antiplasmodial Activity of 80% Methanolic Extract and Solvent Fractions of Stem Bark of Acacia tortilis in Swiss Albino Mice.

Malarial infection has significant negative impact on the health of the world population. It is treated by modern and traditional medicines. Among traditional medicinal plants, Acacia tortilis is used by different communities as antimalarial agent. Therefore, the objective of this study is to validate antimalarial activity of the stem bark of Acacia tortilis in mice. To evaluate antimalarial activity of the plant, 4-day suppressive, curative, and prophylactic antimalarial test models were used. Parasitemia, packed cell volume (PCV), survival time, rectal temperature, and body weight were used to evaluate the effect of the plant extracts. Data were analyzed using SPSS version 26 followed by Tukey's post hoc multiple comparison test. The crude extract and dichloromethane fraction significantly suppressed the level of parasitemia (p < 0.001) and increased mean survival time (p < 0.01) at all tested doses. Similarly, significant effects were observed in mean survival time, % change of PCV, weight, and temperature in both curative and prophylactic antimalarial test models. The methanolic extract and solvent fractions of the stem bark of Acacia tortilis has shown antimalarial activity, and the finding supports the traditional use and the in vitro studies. Thus, this study can be used as an initiation for researchers to find the most active phytochemical entity and to conduct additional safety and efficacy tests.

Leaf Extracts of Cistus ladanifer Exhibit Potent Antioxidant and Antiproliferative Activities against Liver, Prostate and Breast Cancer Cells

Chemical composition, antioxidant, and antiproliferative properties of C. ladanifer crude extracts, including hexane (Hex), dichloromethane (DCM), ethyl acetate (E.A) and ethanol (EtOH) were investigated. The chemical composition of C. ladanifer crude extracts was determined by use of GC-MS, whereas DPPH and FRAP assays were employed to determine its antioxidant capacity. The obtained results showed that the ethanolic extract exhibited a significant antioxidant effect recording an IC50 value of 266.6 ± 0.828 μg/mL with DPPH assay, and a higher reducing power 0.494 ± 0.035 using the FRAP test. The extracts exhibited significant antiproliferative activity against three cancer cell lines. The DCM extract exhibited the highest total polyphenol content (76.066 ± 9.978 μg AGE/mg) and was revealed to be more effective against HepG2 (31.54 ± 0.242 μg/mL). The Hex extract that presented the highest flavonoid content (50.209 ± 3.805 μg CE/mg) exhibited the highest antiproliferative activity against 22Rv1 and MDA-MB-231 recording IC50 values 11.32 ± 2.126 μg/mL and 82.4 ± 1.124 μg/mL, respectively. All four extracts exhibited minimal toxicity against human skin-derived fibroblast cells indicating the specificity of their observed anticancer activity. GC-MS analysis identified interesting phytochemicals underlying the obtained antioxidant and cytotoxic activities. Taken together, results of the current study highlight the significance of C. ladanifer as a valuable source of antioxidant and anticancer bioactive compounds, thereby warranting further detailed investigation.

Coumarins from the Roots of Angelica archangelica and Antibacterial Activity Against Methicillin Resistant Staphylococcus aureus

Angelica archangelica (Fam. Apiaceae) roots were extracted sequentially with n-hexane, dichloromethane (DCM) and methanol (MeOH) using a Soxhlet apparatus. The n-hexane and DCM extracts were fractionated by Vacuum Liquid Chromatography (VLC) over silica gel followed by Preparative Thin Layer Chromatography (PTLC) to yield two coumarins, osthol (1) and osthenol (2). However, the methanol extract was fractionated by solid-phase extraction using C18 cartridge followed by analysis of components by analytical High Performance Liquid Chromatography (HPLC) and purification by preparative High Performance Liquid Chromatography (HPLC) to give two coumarins, bergapten (3) and heraclenol (4) along with sucrose (5). Compounds were identified by a series of 1D (1H and 13C) and 2D (COSY, HSQC and HMBC) NMR spectroscopy and mass spectrometric analysis. The crude extracts (n-hexane, DCM, and methanol) and compound 1, were screened for antimicrobial activity against Gram-positive bacteria (Methicillin resistant Staphylococcus aureus, MRSA and Micrococcus luteus), Gram-negative bacteria, Pseudomonas aeruginosa, and a fungus, Candida albicans. The antimicrobial assays showed that the crude DCM and n-hexane extracts, alongside compound 1, largely inhibited the growth of the microorganisms at high concentrations. Compound 1, which was further tested for antibacterial activity against a series of clinical isolates of methicillin resistance Staphylococcus aureus, showed moderate level of activity against only one strain. Dhaka Univ. J. Pharm. Sci. 20(3): 275-281, 2022 (June) Centennial Special Issue

α-glucosidase inhibition activities of crude extract and mitragynine from Mitragyna Speciosa Korth

The capability in holding the activities of α-glucosidase (1U/mL) of crude extract and mitragynine from Mitragyna speciosa Korth 10 mg/mL. Crude ethyl acetate extracts 61.52±0.47% (IC50 = 17.28 mg/mL) which closed to acarbose 78.94±2.97% (IC50 = 15.74 mg/mL). Minimum density acarbose 0.1562 mg/mL, 4.95±5.96%, crude hexane extracts 2.5 mg/mL, 0.78±11.52%, crude dichloromethane extracts 2.5 mg/mL, 4.40±7.73%, crude ethyl acetate extracts 0.3125 mg/mL, 0.51±5.20 %, crude ethanol extracts 1.25 mg/mL, 0.29±2.54%, crude methanol extracts 0.625 mg/mL, 0.71±0.49%, fractions c (Mitragynine) 5 mg/mL, 1.54±4.50%, crude water extracts, fractions a and b, it was found that there was no activity holding of glucosidase enzyme which meant that korth had the ability to hold the activity of α-glucosidase enzyme in extracting by ethyl acetate. It was also another treatment option and to reduce adverse effects from medication in patients with type 2 diabetes. According to research, the effects of consuming korth leaves should be cautious. Studies on the neuroprotective effects of mitragynine showed that mitragynine could stimulate the nervous system just like cocaine, thereby causing nausea and vomiting. Additionally, korth leaf extract had been reported to be toxic to the liver and kidneys.

Constituents from ripe figs of Ficus vallis-choudae Delile (Moraceae) with antiplasmodial activity.

Ripe figs, barks, and wood of Ficus vallis-choudae are used in traditional medicine against several conditions including nausea and malaria. However, its use is still to be scientifically documented and validated. Hence, the aim of the present work was to evaluate the antiplasmodial activity of the dichloromethane-methanol (DCM-MeOH (1:1)) crude extract, their hexane, dichloromethane, ethyl acetate, and methanoli fractions, as well as the isolated chemical constituents. The chemical study of the DCM-MeOH (1:1) crude extract of F. vallis-choudae figs led to the isolation of fifteen (15) known compounds identified based on their spectroscopic data [one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR), mass spectrometry] and by comparison of these data with those reported in the literature. Some of the isolated compounds were assessed in vitro for their antiplasmodial activity against Plasmodium falciparum chloroquine-sensitive 3D7 (Pf3D7) and multidrug-resistant Dd2 strains. The dichloromethane fraction exhibited very good antiplasmodial activity against both strains with IC50 values of 13.86 μg/mL and 8.18 μg/mL, respectively. Among the tested compounds, wighteone (2) was the most active against P. falciparum 3D7 (IC50 = 24.6 ± 1.5 μM) and Dd2 (IC50 = 11.9 ± 2.4 μM) strains. The obtained results could justify the traditional uses of F. vallis-choudae against malaria. Wighteone appears to be the most active ingredient. However, further consideration of this compound as starting point for antimalarial drug discovery will depend upon its selectivity of action towards Plasmodium parasites.Highlights• 15 (fifteen) compounds were isolated from the dichloromethane-methanol extract of Ficus vallis-choudae.• Their structures were determined on the basis of their spectroscopic data.• The dichloromethane fraction showed promising activities on the Pf3D7 and PfDd2 strains with IC50 values of 13.86 and 8.18 µg/mL, respectively.• Wighteone was the most active compound against PfDd2 (IC50 = 11.9 ± 2.4 μM).Graphical abstract Supplementary InformationThe online version contains supplementary material available at 10.1007/s00436-022-07540-5.

In Vivo and In Vitro Evaluation of Preventive Activity of Inflammation and Free Radical Scavenging Potential of Plant Extracts from Oldenlandia corymbosa L.

Aims: The present study evaluates the anti-inflammatory and antioxidant activity of the crude dichloromethane (CDCME), ethyl acetate (CEAE), and methanol (CMeE) extracts from the plant Oldenlandia corymbosa L. Background:Oldenlandia species have been popular among the people of the Indian subcontinent to treat several types of internal and external inflammation for a long time. Plant decoctions have been used to battle inflammation in cases of tonsilitis, pneumonia and cholecystitis, among others. Objective: The present work designed to demonstrate the properties of the previously mentioned plant extracts to prevent inflammation both in vivo and in vitro. This work is the first investigation of such extracts from this species and their relationship with anti-inflammatory activity. Method: The anti-inflammatory properties of the Oldenlandia corymbosa L. extracts were evaluated in vitro with the Red Blood Cell (RBC) membrane stabilization method and the protein denaturation method and in vivo with the carrageenan-induced paw oedema method. Furthermore, the free radical scavenging activity of the extracts was carried out with the 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical oxidation, total antioxidant capacity and iron reduction assay. Result: Both in vivo and in vitro studies showed that CDCME had the most predominant effects with the value of 80.5% for RBC membrane stabilization, 60% for inhibition of protein denaturation at the concentration of 1000 µg/mL and 63.28% (after 3 h, * p &lt; 0.05) for inhibition of paw oedema (300 mg/kg bwt) compared to carrageenan-induced mice. The free radical scavenging activity was studied by DPPH, total antioxidant and reducing activity assay. CDCME showed scavenging activity in all the methods and an IC50 value of 473.86 µg/mL for DPPH method. Conclusions: The findings of the study remarked that CDCME of the plant has strong anti-inflammatory and antioxidant effects that validate the traditional use of the plant to get remedy from pain. Other: The plants Oldenlandia corymbosa L. were provided by the Bangladesh Council of Scientific and Industrial Research Laboratory campus, Rajshahi, Bangladesh. Experiments on animals were conducted by ethical permission of Institute of Biological Sciences, University of Rajshahi, Bangladesh (license no: 225/320-IAMEBBC/IBSc).

Involvement of Muscarinic Receptors in Hypotensive and Diuretic Effects of Aqueous Soluble Fraction from Asphodelus tenuifolius Cav.

Background. Asphodelus tenuifolius Cav. (Asphodelaceae) is widely used in Pakistan traditional medicine as a hypotensive and diuretic agent. Despite the cardioprotective effects described for A. tenuifolius, the mechanisms involved in its probable hypotensive and diuretic effects have never been evaluated. Firstly, different extracts from A. tenuifolius seeds were obtained, and their antioxidant profiles and chemical constituents by LC-DAD-were determined, including molecular networking by the GNPS platform. Then, to evaluate changes in blood pressure, different groups of anesthetized normotensive rats were intravenously treated with the crude extract (AT-Cr, 1–50 mg/kg), aqueous (AS-AT, 1–25 mg/kg), n-butanol (BS-AT, 1–50 mg/kg), and dichloromethane fraction (DS-AT, 1–80 mg/kg). The diuretic effects of AT-Cr, AS-AT, BS-AT, and DS-AT at 100, 200, and 300 mg/kg, p.o. doses, were also evaluated in comparison with hydrochlorothiazide (HCTZ, 10 mg/kg, p.o). The urinary volume, sodium, potassium, and pH were estimated in the sample collected for 6 h from saline-loaded rats. Using pharmacological antagonists or inhibitors, we determine the involvement of acetylcholine, prostaglandins, and nitric oxide in A. tenuifolius-induced hypotensive and diuresis action. In addition, the activities of angiotensin-converting enzyme, erythrocyte carbonic anhydrase, and renal Na+/K+/ATPase were evaluated in vitro. Acute treatment with crude extract and fractions of A. tenuifolius exhibited significant hypotensive and diuretic potential in normotensive rats. However, AS-AT produced the most potent and significant dose-dependent hypotension and diuretic effects in normotensive rats. Previous treatment with atropine significantly reduced the hypotensive and diuretic action of AS-AT, but pretreatment with indomethacin or L-NAME did not affect these effects. Moreover, the 7-day treatment with AS-AT did not reduce activities of serum angiotensin-converting enzyme, erythrocyte carbonic anhydrase, and renal Na+/K+/ATPase. AS-AT showed four major compound node clusters, which included sugars, alkaloids, nucleoside, amino acid, and glycosylated flavonoids. This research supports and extends the traditional use of A. tenuifolius as a hypotensive and diuretic agent. The results showed that AS-AT from A. tenuifolius could present compounds responsible for hypotensive and diuretic activities through the activation of muscarinic receptors.

Phytochemical and Biological Studies of Bark Extract of Miliusa velutina (Dunal) Hook. f. &amp; Thomson

The main objective of the current research was phytochemical and biological studies of the stem bark of Miliusa velutina (Dunal) Hook. f. &amp; Thomson (Annonaceae). Four purified compounds i.e., friedelin, lupeol, β- sitosterone and caffeic acid were isolated by repeated chromatographic separation and purification of M. velutina. The compounds were identified by analysis of NMR spectral data. The crude dichloromethane extract of stem bark of M. velutina (DEMV) along with its Kupchan partitionates i.e., petroleum ether (PESF), ethyl acetate (EASF), chloroform (CSF) and aqueous (AQSF) soluble fraction were screened for antioxidant, cytotoxic, thrombolytic and antibacterial activities. During the antioxidant activity assay, the AQSF revealed maximum activity with IC50 value of 71.67 μg/ml. The cytotoxicity of plant samples was determined by brine shrimp lethality bioassay, where the maximum cytotoxic activity has been observed for EASF (LC90 = 9.01 μg/ml). In the thrombolytic activity test, the crude dichloromethane extract demonstrated significant efficacy with 46.27% inhibition of clot lysis. In antibacterial screening, the CSF exhibited noticeable inhibitory activity against Shigella boydii with the zone of inhibition 15 mm compared to the standard ciprofloxacin (zone of inhibition = 47 mm).&#x0D; Dhaka Univ. J. Pharm. Sci. 19(2): 125-131, 2020 (December)

Assessment of in vitro anthelmintic activity and bio-guided chemical analysis of BRS Boyrá pineapple leaf extracts

Species of the Bromeliaceae are known for their pharmacological actions, including anthelmintic effects. The aim of this study was to investigate the in vitro anthelmintic activity of extracts and fractions of BRS Boyrá pineapple leaf against the eggs and infective larvae of gastrointestinal nematodes (Trichostrongylidae) of goats and to identify the compounds involved in this activity. Crude methanol, hexane, dichloromethane, ethyl acetate and residual hydromethanol extracts were investigated by quantitative analysis of phenolic and flavonoid contents, antioxidant activity, anthelmintic activity against gastrointestinal nematodes of goats. The extracts were submitted to chromatographic methods for substance isolation and spectrometric techniques to identify their structures. The anthelmintic activity was performed by in vitro assays with eggs and larvae of nematodes obtained from naturally infected donor goats. All extracts contained phenolic (2.22–14.12 g of gallic acid equivalent per 100 g of dry extract) and flavonoid compounds (0.13–1.45 g of quercetin equivalent per 100 g of dry extract). Bio-guided fractionation of the BRS Boyrá pineapple leaves showed high antioxidant activity (EC50 for DPPH of 2.16–21.38 mg mL−1 and inhibition of co-oxidation of β-carotene of 36.40–74.86%) and anthelmintic activity (15.69–100% inhibition of egg hatching). The ethyl acetate extract exhibited greatest activity in all assays. Through chromatographic column analysis it was possible to isolate three substances: β-sitosterol and stigmasterol mixture in dichloromethane and hexane extracts, identified by NMR and p-coumaric acid in the ethyl acetate extract, identified by HPLC-DAD. The isolated p-coumaric acid exhibited high ovicidal effect against goat gastrointestinal nematodes (IC50: 0.12 mg mL−1) and can be considered the active substance of the ethyl acetate extract. This study revealed for the first time that the pineapple BRS Boyrá possesses inhibitory activity against gastrointestinal nematodes (Haemonchus spp., Oesophagostomum spp. and Trichostrongylus spp.), and that p-coumaric acid is an important bioactive.

Immunomodulatory potential of extracts, fractions and pure compounds from Phyllanthus amarus and Psidium guajava on striped catfish (Pangasianodon hypophthalmus) head kidney leukocytes

This study aimed to identify major phytochemical constituents, as well as compare the immunomodulatory effects of Psidium guajava L. and Phyllanthus amarus Schun and Thonn crude ethanol extracts and their fractions on striped catfish (Pangasianodon hypophthalmus) head kidney leukocytes (HKLs). Moreover, pure constituents were also investigated for their effects on those cells: hypophyllanthin, identified as a major constituent of P. amarus crude extracts and its hexane fraction; corosolic acid, ursolic acid, and oleanolic acid, identified in P. guajava crude extract, ethyl acetate and dichloromethane fractions; with other terpenic derivatives, as well as guajaverin and avicularin, identified with other flavonoids by LC-UV-MS in the crude P. guajava extract and its ethyl acetate fraction. Cell viability, respiratory burst assay (RBA), nitric oxide synthase (NOS) and lysozyme activity in HKLs were analyzed after 24 h stimulation with each extract (10, 20 and 40 μg/mL) or pure compound (7.5, 15 and 30 μM). Our results show that the hexane fraction of both plant extracts inhibited the viability of HKLs, while several other fractions enhanced the cell viability. All P. guajava fractions at all or some concentration considerably enhanced the RBA production in HKLs. Similarly, NOS production was also significantly increased by some or all concentrations of P. guajava dichloromethane and ethyl acetate fractions. However, the NOS production was dose-dependently inhibited in HKLs treated with Pa ethyl acetate and both plants aqueous fractions at 10 or 10 and 40 μg/mL respectively. The lysozyme activity in cells treated with P. guajava crude extracts and all its organic solvent fractions were stronger than those in P. amarus treatments. Pure compounds including corosolic acid, guajaverin, ursolic acid, hypophyllanthin inhibited the HKLs viability according to concentration and type of compound. All pure compounds except avicularin significantly stimulated, at certain or all concentrations, the RBA production and/or the lysozyme activity in HKLs. The NOS production was significantly reduced in HKLs treated with oleanolic acid (30 μM) and hypophyllanthin (7.5 μM) while its level was increased by hypophyllanthin at 30 μM. These results highlighted that the crude ethanol extracts of P. guajava and P. amarus, their fractions and some of their pure components at certain concentrations can potentially act as immunomodulators, and could be considered as valuable candidates in fishery sciences.