Dichloromethane Soluble Fraction Research Articles

Phytochemical Screening, Antioxidant, Cytotoxic, Analgesic, Antidiarrheal Activity and GC-MS Analysis of the Leaves of Stachyphrynium Placentarium

The goal of the present study is to analyze the phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of the leaves of Stachyphrynium placentarium methanol extract as well as its dichloromethane soluble fraction using GC-MS technique. The leaves of the S. placentarium were extracted with methanol and designated as SPM. Phytochemicals investigation exhibited the presence of several important secondary metabolites. Antioxidants such as total phenolic content, flavonoids content, total antioxidant, reducing power capacity and DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay SPM revealed non-significant antioxidant properties. The cytotoxicity study of the extract showed significant cytotoxicity (LC50 35.1093 µg/mL) compared to standard vincristine sulfate (LC50 0.482 µg/mL). Analgesic study of S. placentarium by hot plate model indicates both the doses exhibited significant central analgesic activity compared with the control. The maximum analgesic activity was observed at 90 min. SPM (500 mg/kg) is 60.21 %. SPM250 also showed a significant percentage of elongation or latency from 30 to 150 min 57.00, 59.64, 59.88, 58.07 and 53.71 %, respectively 30 min time interval. In the acetic acid writhing test, S. placentarium significantly outperformed the control and standard (SPM250 - 52.27 %, SPM500 - 34.09 and STD - 27.27 %), showing a significant increase in peripheral analgesic activity (p < 0.001). Antidiarrheal activity of SPM showed that SPM250 and SPM500 substantially decreased the feces in castor oil-induced diarrhea (p < 0.05). According to the study, the incidence and severity of diarrhea are significantly reduced by SPM500. Four chemicals are found in the dichloromethane-soluble portion of the methanol extract of S. placentarium leaves, according to GC-MS studies such as Octadecane, 6-methyl- (1.326 %), Dodecane (1.02 %), 1-Iodo-2-methylundecane (0.47 %), tetradecanoic acid 12-methyl, methyl ester (72.64 %). Tetradecanoic acid exhibits anti-inflammatory, anticancer, 5-reductase inhibitory, antioxidant, hemolytic and antifibrinolytic properties. HIGHLIGHTS The phytochemicals, antioxidants, cytotoxic, analgesic and anti-diarrheal properties of Stachyphrynium placentarium methanol extract (SPM) leaves. The dichloromethane soluble fraction is examined using Gas chromatography–mass spectrometry (GC-MS) and identified Octadecane, 6-methyl-, Dodecane, 1-Iodo-2-methylundecane, tetradecanoic acid 12-methyl, methyl ester. The brine shrimp lethality test exhibited that SPM has significant cytotoxicity (LC50 35.1093 µg/mL) compared to vincristine sulphate (LC50 0.482µg/mL), the extract showed significant cytotoxicity (LC50 35.1093 µg/mL). S. placentarium at both dosages (250 and 500 mg) provided significant central analgesia compared to the control in a hot plate model. In the acetic acid writhing test, S. placentarium showed significantly higher peripheral analgesic action (SPM250 - 52.27, SPM500 - 34.09 and STD - 27.27 %) compared to control and standard. In castor oil-induced diarrhoea, SPM250 and SPM500 (250 and 500 mg) showed significantly reduced faeces production (p < 0.05). GRAPHICAL ABSTRACT

Antioxidant, membrane stabilizing and thrombolytic potentials of the leaves of erythrina fusca lour

Erythrina fusca (Family: Fabaceae) is famous for many traditional healing. In the present study, the leaves of E. fusca were collected, dried, powdered and then extracted with methanol. It was further partitioned into n-hexane, dichloromethane and aqueous soluble fractions. All the extracts were subjected to antioxidant, membrane stabilizing and thrombolytic assays. In case of free radical scavenging assay, the aqueous soluble fraction showed highest scavenging activity. In ferric reducing anti-oxidant power assay, the dicholoromethane extract showed highest reduction of ferric ion in a dose dependent manner. In total antioxidant assay, the methanol extract and dichloromethane extract showed stronger capacity as antioxidant. During the determination of total phenolic and flavonoid contents, the methanol extract and dichloromethane soluble fraction showed higher contents of phenolics and flavonoids, respectively. The erythrocyte membrane stabilizing assay revealed the capacities of methanol extract and its aqueous fraction to stabilize the RBC membrane in both hypotonic solution- and heat- induced hemolytic conditions. However, the clot lysis ability of the E. fusca leaf extracts was not so prominent. Bangladesh J. Bot. 53(3): 439-445, 2024 (September)

Bioassay-directed isolation of antiplasmodial alkaloids and lignans from the dry seeds of Piper guineense Schumach. & Thonn. (Piperaceae) and their cytotoxicity

The identification of antiplasmodial lead compounds from the dry seeds of Piper guineense has been performed using a bioassay-directed protocol. Briefly, its methanolic seeds extract has been suspended in water and partitioned using n-hexane, dichloromethane, ethyl acetate and n-butanol to afford four main fractions A−D which were further submitted to antiplasmodial assay against Plasmodium falciparum 3D7. The bioassay results showed that its dichloromethane-soluble fraction has the highest potency with 31.8 % of the parasite P. falciparum 3D7 survival. The purification of the active fraction with successive column chromatography led to the isolation and identification of eleven compounds including six piperine-type amides (1−6), two lignans (7 and 8), one fatty acid (9), and two steroids (10 and 11). The antiplasmodial assay of compounds 1−8 against the two strains Pf 3D7 and Pf Dd2 showed that only the alkaloids 1−6 displayed moderate potencies with the highest IC50 values of 17.4 ± 0.7 µg/mL (for 6 against Pf 3D7) and 14.6 ± 0.3 µg/mL (for 2 against Pf Dd2). Furthermore, the cytotoxicity of the tested compounds against Vero cell lines showed that only compounds 5 and 6 displayed a cytotoxicity with CC50 values of 335.4 ± 7.6 µg/mL and 151.8 ± 2.4 µg/mL, respectively, while their selectivity index limits an acceptable range. The results support the use of the plant in folk medicine for handling malaria, and demonstrate that piperine-type amides are responsible for the antiplasmodial activity of P. guineense and are good leads in the development of safe and potent antimalarial drugs.

Pharmacological Profile Analysis of a Vegetable Crop Pumpkin (Cucurbita maxima Linn.) Seeds

The seeds of pumpkin (Cucurbita maxima Linn.) can be utilized as both a conventional and multifunctional dietary constituent. In this study the antioxidant, antibacterial, and cytotoxic effects of various fractions such as petroleum ether soluble fraction (PESF), chloroform soluble fraction (CFSF), dichloromethane soluble fraction (DCMSF), and aqueous soluble fraction (AQF) were obtained from crude extract (CE) of the C. maxima seeds were assessed by using 1,1-Diphenyl-2- Picrylhydrazyl (DPPH) method, disc diffusion technique, and brine shrimp lethality bioassay respectively. The results showed that all the fractions have remarkable antioxidant, antibacterial, and cytotoxic activities. The study revealed that both the AQF and PESF and the CF confirmed significant antioxidant properties. The antibacterial activity of different fractions against Gram-positive and Gram-negative bacteria was examined to determine their significant effect. PESF and CFSF fractions exhibited substantial inhibitory activity against Gram-positive bacteria, particularly Bacillus cereus. Bacillus subtilis exhibited significant susceptibility to both CFSF and AQF. Salmonella typhi and Shigella dysenteriae exhibited significant susceptibility to PESF and CFSF, while Escherichia coli was the Gram-negative bacterium most effectively inhibited by CFSF. The IC50 values revealed that the DCMSF exhibited the highest inhibitory activity, followed by other fractions. The new study emphasizes the significance of pumpkin seeds in terms of their pharmacological and biological properties, which may encourage more research on the potential benefits of this plant for human health. Jagannath University Journal of Life and Earth Sciences, 9(1): 7-18, 2023 (June)

Multifaceted Chemico-Pharmacological Insights into Cynometra ramiflora L.: Unveiling its GC-MS, Cytotoxic, Thrombolytic, Anti-Inflammatory, Antioxidant, Anti-Diarrheal, Hypoglycemic, and Analgesic Potentials

Objective: This study aimed to comprehensively evaluate the chemico-pharmacological properties of methanolic extractives derived from Cyanometra ramiflora leaves, encompassing a spectrum of activities including cytotoxic, thrombolytic, membrane-stabilizing, antioxidant, antidiarrheal, hypoglycemic, and analgesic effects. Methods: Phytomolecules were identified using Gas Chromatography-Mass Spectrometry (GC-MS), while antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and total phenolic content tests. Cytotoxicity was determined through the brine shrimp bioassay, while thrombolytic activity was evaluated via clot lysis assays, and membrane-stabilizing effects were investigated for potential anti-inflammatory properties. In-vivo experiments using mice were conducted solely with the methanolic extract to assess central and peripheral analgesic, hypoglycemic, and antidiarrheal properties. Results: The presence of 47 compounds in the methanolic extract, with tetratriacontyl pentafluoropropionate (9.25%), pentatriacontane (8.75%), and 2-methylhexacosane (5.9%) being predominant. The dichloromethane-soluble fraction exhibited the highest cytotoxicity, with a 50% lethal concentration (LC50) of 11.05 µg/mL. Both petroleum-ether and methanol-soluble fractions demonstrated more than 50% lysis in thrombolytic activity assays. Under hypotonic and heat-induced conditions, the ethyl acetate and dichloromethane-soluble fractions of C. ramiflora significantly reduced erythrocyte hemolysis. Notably, the methanol-soluble fraction exhibited significant antioxidant potential with 50% inhibitory concentration (IC50) of 17.71 μg/mL compared to the standard EDTA (IC50 = 75.94 μg/mL). The hypoglycemic investigation revealed a 63.73% reduction at 90 min with a 600 mg/kg dose, while mild antidiarrheal activity was observed with a 37.5% reduction in diarrhea at 4 h intervals. Significant analgesic effects were demonstrated in both central and peripheral tests, with a 68.53% reduction in writhing during the peripheral test and an average tail immersion time of 5.38 s at 90 min. Conclusion: C. ramiflora leaves show promise as natural sources for various pharmacological activities, including cytotoxicity, thrombosis, inflammation, antioxidant, hypoglycemic, and analgesic effects. However, further comprehensive research is recommended to elucidate the precise mechanisms of action and analyze the phytochemical composition of C. ramiflora plants.

Influence of the flowering stage in the production of urechitol A in Agrobacterium rhizogenes-transformed plants of Pentalinon andrieuxii

Background: Pentalinon andrieuxii, is a medicinal plant used in Mayan folk medicine against leishmaniosis. It is an important source of terpenoids whose biosynthesis has been reported to be influenced by the ontogeny of the plant.
 Hypotheses: Flowering positively influences the production of urechitol A in Pentalinon andrieuxii.
 Studied species: Pentalinon andrieuxii (Müll. Arg.) B.F. Hansen & Wunderlin (Apocynaceae)
 Study site and dates: Mérida, Yucatán, Mexico, 2019 and 2020.
 Methods: Flowering was induced by exposing plants to temperatures above 32 °C. Plant tissue samples were collected before flowering induction and at full bloom and extracted with methanol. Analyses of the dichloromethane-soluble fraction of the crude extract by Gas Chromatography-Mass Spectrometry allowed the detection and quantification of urechitol A.
 Results: Flowering stage of P. andrieuxii has a positive influence on the production of urechitol A in the root of the plant, with the contents of the tri-nor-sesquiterpene increasing from a minimum of 43 to a maximum of 91 times during the flowering stage of plants, both transformed and wild type.
 Conclusions: P. andrieuxii perceives high temperatures as an important environmental cue to flower; flowering positively influences the production of urechitol A in the roots of the plant; production of the tri-nor-sesquiterpene is controlled by the plant.

Chemico-pharmacological and computational studies of Ophiorrhiza fasciculata D. Don and Psychotria silhetensis Hook. f. focusing cytotoxic, thrombolytic, anti-inflammatory, antioxidant, and antibacterial properties

The current study sought to examine the pharmacological potentials of crude methanolic extracts of Ophiorrhiza fasciculata and Psychotria silhetensis, as well as their various solvent fractionates, with a focus on cytotoxic, thrombolytic, membrane stabilizing, antioxidant, and antibacterial activities via in vitro and in silico approaches. The extensive chromatographic and spectroscopic analyses confirmed and characterized two compounds as (±)-licarin B (1) and stigmasterol (2) from O. fasciculata and P. silhetensis, respectively. Petroleum ether soluble fraction of O. fasciculata and the aqueous soluble fraction of P. silhetensis showed the lowest 50% lethal concentrations (1.41 and 1.94 μg/mL, respectively) in brine shrimp bioassay. Likewise, petroleum ether soluble fraction of O. fasciculata and aqueous soluble fraction of P. silhetensis showed the highest thrombolytic activity with 46.66% and 50.10% lyses of the clot, respectively. The methanol and dichloromethane soluble fractions of O. fasciculata reduced erythrocyte hemolysis by 64.03% and 37.08%, respectively, under hypotonic and heat-induced conditions, compared to 81.97% and 42.12% for standard acetylsalicylic acid. In antioxidant activity test, aqueous soluble fraction O. fasciculata (IC50 = 7.22 μg/mL) revealed promising antioxidant potentialities in comparison to standard butylated hydroxytoluene (IC50 = 21.20 μg/mL). In antibacterial screening, chloroform, and dichloromethane soluble fractions of P. silhetensis showed a mild antibacterial activity compared with the standard drug ciprofloxacin. Additionally, the molecular docking study corroborated the current in vitro findings, and the isolated two constituents had higher binding affinities toward epidermal growth factor receptor, tissue plasminogen activator, vFLIP-IKK gamma stapled peptide dimer, glutathione reductase, and dihydrofolate reductase enzyme than their corresponding standard drugs. In addition, the both isolated compounds exerted favorable pharmacokinetics (absorption, distribution, metabolism, excretion) and toxicological profiles with drug-like qualities in computational-based ADMET and drug likeliness analyses. The current research suggests that both plants have potential as a natural treatment for treating thrombosis, inflammation, and oxidative stress. However, more thorough research is required to thoroughly screen for phytochemicals and pinpoint the precise mechanisms of action of the bioactive metabolites derived from these plants against a broad range of molecular targets.

Evaluation of Antioxidant, Cytotoxic, Thrombolytic and Membrane Stabilizing Activities of Canna indica L. Leaves (Family: Cannaceae)

Canna indica L., a perennial plant known as Indian shot and African arrowroot, is a member of the family Cannaceae. Historically, this plant was used to treat menstrual cramps, diarrhea and dysentery. The purpose of this investigation was to assess the antioxidant, cytotoxic, thrombolytic and membrane stabilizing activities of the methanol extract of C. indica leaf and its organic soluble fractions. The DPPH free radical scavenging method and the total phenolic content determination were utilized to evaluate the antioxidant capacity, whereas the brine shrimp lethality bioassay was implemented to study the cytotoxic potential. The ability of the extractives to prevent the hemolysis of red blood cells (RBC) in response to the application of heat and hypotonic solutions was used to estimate of their membrane stabilizing potential. In all of these experiments, the methanolic leaf extract (ME) displayed moderate activity, while the petroleum ether soluble fraction (PSF) and the dichloromethane soluble fraction (DSF) demonstrated the most promising antioxidant and cytotoxic activities among the extractives. However, the strongest thrombolytic and membrane stabilizing activities were observed by the chloroform soluble fraction (CSF). Additional research may be necessary to determine which phytoconstituents are most likely to be responsible for the aforementioned qualities. Bangladesh Pharmaceutical Journal 26(2): 162-166, 2023 (July)

C21 steroids from the roots of Marsdenia tenacissima

A phytochemical investigation of the dichloromethane soluble fraction of the ethanolic extract obtained from the roots of Marsdenia tenacissima led to the discovery of the sixteen undescribed pregnane C21 steroids (1–16) and isolation of eleven known C21 steroidal analogues (17–27). Their chemical structures were elucidated by one- and two-dimensional nuclear magnetic resonance spectroscopy and, high resolution–electrospray ionization mass spectrometry and their absolute configurations were determined using electronic circular dichroism or single-crystal X-ray diffraction. The in vitro anti-proliferative effects of 1–16 were evaluated against HepG2 (human hepatocellular cancer), A549 (lung cancer), and MCF-7 (human breast cancer) cell lines. Even though some of them showed moderate cytotoxic activities, marsectohexol derivative 12 exhibited significant cytotoxicity against A549 cells with an IC50 value of 5.2 μM.

Bioassay-Guided Isolation of Antiplasmodial Compounds from Hypericum lanceolatum Lam. (Hypericaceae) and Their Cytotoxicity and Molecular Docking.

In Cameroon, malaria is still the cause of several deaths yearly and leading to the continued search for new potent leads to fight against Plasmodium falciparum. Medicinal plants like Hypericum lanceolatum Lam. are introduced in local preparations for the treatment of affected people. The bioassay-guided fractionation of the crude extract of the twigs and stem bark of H. lanceolatum Lam. led to the identification of the dichloromethane-soluble fraction as the most active (with 32.6% of the parasite P. falciparum 3D7 survival) which was further purified by successive column chromatography to obtain four compounds identified by their spectrometric data as two xanthones 1,6-dihydroxyxanthone (1) and norathyriol (2) and two triterpenes betulinic acid (3) and ursolic acid (4). In the antiplasmodial assay against P. falciparum 3D7, the triterpenoids 3 and 4 displayed the most significant potencies with IC50 values of 2.8 ± 0.8 μg/mL and 11.8 ± 3.2 μg/mL, respectively. Furthermore, both compounds were also the most cytotoxic against P388 cell lines with IC50 values of 6.8 ± 2.2 μg/mL and 2.5 ± 0.6 μg/mL, respectively. Further insights on the inhibition method of the bioactive compounds and their drug-likeness were obtained from their molecular docking and ADMET studies. The results obtained help in identifying additional antiplasmodial agents from H. lanceolatum and support its use in folk medicine for the treatment of malaria. The plant might be considered as a promising source of new antiplasmodial candidates in new drug discovery.

Bioguided isolation of potential antitumor agents from the aerial parts of cultivated cardoon (Cynara cardunculus var. altilis)

Hepatocellular carcinoma (HCC) is one of the leading causes of mortality worldwide; therefore, searching for an effective treatment for this illness is of great importance. In the present work,in vitrocytotoxic activity of the ethanol extract of the aerial parts ofCynara cardunculusL. against human liver carcinoma cells (Hep G2) was tested. Additionally, the antitumor activity of the extract was confirmed using chemically induced rat liver carcinogenesis with diethylnitrosamine (DEN). Moreover, bioguided fractionation and column chromatographic separation of the active compounds were carried out. The extract of C. cardunculus showed a promising cytotoxic activity according to the protocols of the National Cancer Institute. Bioguided chromatographic separation of the ethanol extract of C. cardunculus led to the isolation of seven secondary metabolites including two sesquiterpene lactones as the principal active components of the methylene chloride soluble fraction, grosheimin (IC50 = 7.49 µg/mL) and cynaropicrin (IC50 = 13.9 µg/mL). The compounds were characterized by different spectroscopic techniques such as EI-MS, IR and NMR. Additionally, in silico analysis of the two active compounds revealed their ability to bind with caspase-3 via hydrogen bonds interactions to initiate apoptosis of cancer cells. The results shed the light on the significance of C. cardunculus as a potential source of antitumor agents.

Phenolic Constituents from Wendlandia tinctoria var. grandis (Roxb.) DC. Stem Deciphering Pharmacological Potentials against Oxidation, Hyperglycemia, and Diarrhea: Phyto-Pharmacological and Computational Approaches.

Wendlandia tinctoria var. grandis (Roxb.) DC. (Family: Rubiaceae) is a semi-evergreen shrub distributed over tropical and subtropical Asia. The present research intended to explore the pharmacological potential of the stem extract of W. tinctoria, focusing on the antioxidant, hypoglycemic, and antidiarrheal properties, and to isolate various secondary metabolites as mediators of such activities. A total of eight phenolic compounds were isolated from the dichloromethane soluble fraction of the stem extract of this plant, which were characterized by electrospray ionization (ESI) mass spectrometric and 1H NMR spectroscopic data as liquiritigenin (1), naringenin (2), apigenin (3), kaempferol (4), glabridin (5), ferulic acid (6), 4-hydroxybenzoic acid (7), and 4-hydroxybenzaldehyde (8). The dichloromethane soluble fraction exhibited the highest phenolic content (289.87 ± 0.47 mg of GAE/g of dried extract) and the highest scavenging activity (IC50 = 18.83 ± 0.07 µg/mL) against the DPPH free radical. All of the isolated compounds, except 4-hydroxybenzaldehyde, exerted a higher antioxidant effect (IC50 = 6.20 ± 0.10 to 16.11 ± 0.02 μg/mL) than the standard butylated hydroxytoluene (BHT) (IC50 = 17.09 ± 0.01 μg/mL). Significant hypoglycemic and antidiarrheal activities of the methanolic crude extract at both doses (200 mg/kg bw and 400 mg/kg bw) were observed in a time-dependent manner. Furthermore, the computational modeling study supported the current in vitro and in vivo findings, and the isolated constituents had a higher or comparable binding affinity for glutathione reductase and urase oxidase enzymes, glucose transporter 3 (GLUT 3), and kappa-opioid receptor, inferring potential antioxidant, hypoglycemic, and antidiarrheal properties, respectively. This is the first report of all of these phenolic compounds being isolated from this plant species and even the first demonstration of the plant stem extract’s antioxidant, hypoglycemic, and antidiarrheal potentials. According to the current findings, the W. tinctoria stem could be a potential natural remedy for treating oxidative stress, hyperglycemia, and diarrhea. Nevertheless, further extensive investigation is crucial for thorough phytochemical screening and determining the precise mechanisms of action of the plant-derived bioactive metabolites against broad-spectrum molecular targets.

Phyto-pharmacological Investigations of Leaves of Cissus trifoliata (L.)

Repeated chromatographic separation and purification of dichloromethane soluble partitionate of methanol extract of leaves of Cissus trifoliata (L.) yielded a total of four compounds, namely β-sitosterol (1), β- sitostenone (2), epifriedelanol (3) and friedelin (4). The structures of the isolated chemical compounds were established by high field nuclear magnetic resonance (NMR) analyses, comparison with previously published data and co-TLC with authentic samples, whenever possible. Compounds 1-4 appeared to be the first time they were identified in this plant species. The methanol extract of C. trifoliata was assessed for central and peripheral analgesic activities by tail immersion and acetic acid-induced writhing reflex methods, respectively. The crude extract demonstrated significant central and peripheral analgesic efficacy at 200- and 400-mg/kg b.w. in mice model. The general toxicity of the extractives was assessed through a brine shrimp lethality bioassay, where the ethyl acetate (LC50=0.88 μg ml–1) and dichloromethane (LC50=0.93 μg ml–1) soluble fractions demonstrated considerable activity. The crude extract at 400 mg/kg (b.w.) also displayed significant antidiarrheal activity, whereas its dichloromethane soluble fractions exhibited moderate thrombolytic activity. Dhaka Univ. J. Pharm. Sci. 21(1): 69-75, 2022 (June)

Toxicity and growth characteristics of epiphytic dinoflagellate Gambierdiscus silvae in Japan

The genus Gambierdiscus is a marine benthic/epiphytic dinoflagellate that has been investigated worldwide as the causative agent of ciguatera poisoning (CP). In Japan, CP occurs mainly in the subtropical region and sporadically in the temperate region. To understand the mechanism of CP outbreaks in the coastal regions, identifying the species of Gambierdiscus occurring in the regions and determining their toxicity and growth characteristics, such as growth responses to temperature, salinity, and light intensity, are important. Recently, the occurrence of G. silvae in the Japanese temperate and subtropical regions has been revealed through metabarcoding. However, the toxicity and growth characteristics of G. silvae have not yet been investigated. In this study, three strains of Gambierdiscus were isolated from a depth of 30 m in subtropical waters in Japan and were identified as Gambierdiscus silvae based on morphological characteristics and phylogenetic positions. A dichloromethane soluble fraction (DSF) and aqueous methanol soluble fraction (MSF) of the three strains showed high mouse toxicity by intraperitoneal injection, but only the DSF of the three strains showed toxicity by gavage. All strains grew in the range of 17.5–30 °C and salinity range of 25–40, and grew well at 25 °C and salinity 30. The optimal light intensity for growth of the strains was 42.0–83.0 μmol photons/m2/s. These results suggest that G. silvae has the potential to be widely distributed from temperate to subtropical/ regions and in shallow to deep coastal waters of Japan. Understanding the growth characteristics of this species would be useful in predicting the occurrence of this species in Japanese coastal waters. Finally, the results obtained in this study suggest that G. silvae showing high toxicity is one of the causative agents of CP in Japan, and knowledge of this species would be useful in understanding the mechanism of CP outbreaks in Japan.

Lupane Triterpenoids, Phytosterol, Phenolic Acid and Fatty Acid From Leaves of Bouea Oppositifolia (Roxb.) Meisn.

Phytochemical investigation of petroleum ether (PESF) and dichloromethane soluble (DCMSF) fractions designated as PESF and DCMSF, respectively of methanol extract of leaves of Bouea oppositifolia (Roxb. Meisn. (an endangered species of Bangladesh) yielded six compounds, which were characterized as betulin (1), lupeol (2), lupenone (3), stigmasterol (4), p-coumaric acid (5), and oleic acid (6) by high field NMR studies and comparison with published values. The identity of the isolated compounds were further substantiated by co-TLC. Bangladesh J. Bot. 51(1): 185-189, 2022 (March)

Diterpenes and sterols from the Red Sea soft coral Sarcophyton trocheliophorum and their cytotoxicity and anti-leishmanial activities

The ethyl acetate and dichloromethane-soluble fractions, from a soft coral Sarcophyton trocheliophorum total methanolic extract, exhibited significant anti-leishmanial and cytotoxic activities. These active fractions yielded a new cembranoid diterpene (1), two known analogues [sarcotrocheliol (2) and sarcophine (3)], and two sterols [(24S)-24-methylcholesterol (4) and gorgosterol (5)]. The structure of the new diterpene (1) was determined via a detailed analysis of its spectroscopic data. Compounds 3 and 5 demonstrated noticeable cytotoxicity on A549 (IC50 17.4 ± 1.9 µg/ml) and HepG2 (IC50 17.7 ± 1.5 µg/ml) cell lines, respectively. None of the isolates 1‒5 showed detectable anti-leishmanial activity (IC50 >100 µg/ml).

In vitro cytotoxic and membrane stabilizing activities of Alternanthera versicolor L.

The crude methanolic extract of leaves of Alternanthera versicolor L. as well as their pet-ether, carbon tetrachloride, chloroform and aqueous soluble fractions were tested to investigate the membrane stabilizing and cytotoxic activity. Among all extractives, dichloromethane soluble fraction (DCMSF) of crude extract of leaves of Alternanthera versicolor L. exhibited lowest cytotoxicity effect with LC50 value 30.63 µg/ml. From investigation results it can also be seen that mortality increased gradually with an increase in concentration of the test samples and maximum mortalities took place at the highest concentration of 400µg/ml. In addition, aqueous soluble fraction (AQSF) shows highest % inhibition of hypotonic solution induces hemolysis of erythrocyte membrane and dichloromethane soluble fraction (DCMSF) shows highest % inhibition of heat induces hemolysis of erythrocyte membrane. Therefore, considering the potential bioactivity, the plant can further be studied extensively to find out their unexplored efficacy and to rationalize their uses of traditional medicine.

In vivo bioactivity-guided isolation of antifertility fraction of Waltheria indica Linn. root in male Wistar rats

BackgroundWaltheria indica is a multipurpose medicinal plant with abundance of phytochemical compounds. Antifertility effect of Waltheria indica Linn. root and leaves have been reported. However, the fraction responsible for this antifertility effect needs to be isolated for possible male contraceptive purpose. Therefore, this research was designed to isolate the antifertility fraction of Waltheria indica Linn. root (WILR) in an in vivo model using male Wistar rats. Crude ethanol extract of WILR was sequentially dissolved in hexane, dichloromethane, and ethyl acetate. Rats (n = 5) were administered with 200, 500, or 1000 μg/kg of hexane, dichloromethane, and ethyl acetate soluble extracts of WILR, while control received distilled water, daily for 15 days to determine the soluble extract with most antifertility effect. Thereafter, fractions were separated from dichloromethane soluble WILR extract by column and thin-layer chromatography. Rats (7 groups, n = 5) were administered with each of the fractions (DF1 to DF7; at 1000 μg/kg) to determine the fraction with the highest antifertility. Rats were thereafter sacrificed, and sperm parameters, reproductive hormones, testicular cholesterol, and protein were determined according to standard procedure. Histology of the testis was also done. Data were analyzed using ANOVA at p ≤ 0.05.ResultsDichloromethane soluble fraction (500 μg/kg) significantly decreased sperm concentration (137.00 ± 9.85 to 107.00 ± 13.08 × 106 cells/mL), levels of testosterone (2.90 ± 0.65 to 1.50 ± 0.37 ng/mL), and FSH (0.08 ± 0.08 to 0.99 ± 0.08 IU/L). The dichloromethane soluble fraction also caused the loss of testicular interstitium and spermatogenic cells. DF5 significantly reduced sperm motility (92.00 ± 2.74 to 76.00 ± 5.48%) and LH (2.86 ± 0.52 to 1.47 ± 0.18 IU/L). DF5 also significantly increased levels of prolactin (1.22 ± 0.10 to 1.88 ± 0.48 ng/mL), testicular total protein (7.36 ± 0.35 to 8.54 ± 1.06 g/dL), and testicular cholesterol (34.17 ± 3.65 to 55.76 ± 6.08 mg/mL).ConclusionThe results indicate that the DF5 is the bioactive fraction of WILR responsible for its antifertility effect. The possible antifertility mechanisms are through the reduction in sperm parameters, reproductive hormones, and histological changes in the testis.

Phytochemical Investigation of Schleichera oleosa (Lour.) Oken Leaf

Schleichera oleosa (Lour.) Oken. is a medicinal plant of Bangladesh with enormous traditional applications in folk medicine. The current study was designed to isolate the secondary metabolites by successive chromatographic separation of n-hexane and dichloromethane soluble fractions of a methanol extract of leaves of S. oleosa. A total of four compounds were separated and identified as 5,7-dihydroxy-4'-methoxyflavone, stigmasterol, lupeol and betulinic acid. The structures of the isolated compounds were elucidated by analysis of their 1H NMR data and comparison with published values. Bangladesh Pharmaceutical Journal 24(1): 33-36, 2021

In vitro and in vivo evaluation of pharmacological potential of Begonia barbata Wall

BackgroundBegonia barbata Wall is an important medicinal plant in Bangladesh used to treat several human diseases. The intention of the existing work was to determine in vitro and in vivo pharmacological properties of petroleum-ether, dichloromethane, and ethyl acetate soluble fractions of methanol extract of B. barbata whole plant (MEBB). The total phenolic content (TPC) was determined by Folin-Ciocalteu test while antioxidant activity assay was performed by DPPH radical quenching process. Tail immersion and acetic acid-induced writhing methods were utilized to evaluate analgesic effect in albino mice. To evaluate the hypoglycemic potential, glucose tolerance test is used. Castor oil-induced diarrhea method is utilized to figure out the anti-diarrheal action in mice. Thiopental-sodium-induced sleeping time test was exploited for the appraisal of CNS antidepressant action.ResultsAntioxidant activities of the test materials followed the order: dichloromethane fraction (DCMF) > ethyl acetate fraction (EAF) > petroleum-ether fraction (PEF). The EAF yielded maximum amount of phenolic compounds (12.47 mg of GAE/g of sample). The DMF was most effective in terms of thrombolytic activity while the EAF protected the membrane integrity of erythrocyte by 63.47% inhibition of hemolysis induced by heat. Among all fractions, the PEF (400 mg/kg body weight) demonstrated highest analgesic activity in both tail immersion test (maximum elongations as 262.053%) and acetic acid-induced writhing method (66.02% writhing inhibition). During anti-diarrheal test, the plant samples significantly diminished the frequency of diarrheal episodes in mice. Similarly, the plant samples decreased the sleeping duration induced by thiopental sodium.ConclusionThe above outcome exposed that B. barbata possesses significant pharmacological potential, which validates its use in Bangladesh folk medicinal practices.