Crude Hexane Extracts Research Articles

Nematicidal efficacy of Cymbopogon nardus, Pelargonium graveolens essential oils and Annona squamosa extract against Meloidogyne incognita and M. graminicola

A crude hexane extract of seed kernel was processed into fatty acid methyl esters (FAME) using methanolic-sulfuric acid. Herbal mixtures (seed extracts of A. squamosa and essential oils of Cymbopogon nardus (citronella) and Pelargonium graveolens (geranium)) were evaluated against root knot nematode (Meloidogyne spp). The mortality of J2s of M. incognita in hexane and methanol extracts (AHE+AME) of A. squamosa with geranium oil (GO) ranged from 50.75-89.75% to 71.50-99.00% at 31.25 to 1000 μgmL-1 in 24 to 96 hours. The number of galls after CO+AHE+AME treatment was 4.0 ± 0.0 and 5.67 ± 0.58 galls/seedling and were recorded at 1000 and 500 μgmL-1 treatments on 30 days after inoculation, compared to carbofuran (3.0 ± 0.0 and 4.3 ± 0.6 galls/seedling) at 1000 and 500 μgmL-1 and velum prime (3.6 ± 0.6 and 4.6 ± 0.6 galls/seedling) at 1000 and 500 μgmL-1. The maximum shoot length of brinjal seedlings was recorded in CO+AHE+AME treatment compared to carbofuran (47.3 ± 0.6cm) and velum prime (48.7 ± 0.6 cm) at 1000 μg mL-1 in soil drenching application. Methanol and hexane extracts (AHE+AME) obtained from Annona squamosa seeds were investigated for their nematicidal properties. M. graminicola in direct-seeded rice at the nursery level and M. incognita in brinjal (eggplant) were controlled for up to sixty days when treated with these mixtures.

α-GLUCOSIDASE AND TYROSINASE INHIBITORY GUIDED ISOLATION, IDENTIFICATION OF MAJOR COMPOUND AND IN SILICO STUDY FROM EDIBLE MUSHROOM COPRINUS COMATUS

Coprinus comatus is an edible and medicinal mushroom. The crude hexane, EtOAc and methanol extracts were extracted successively from C. Comatus. All the crude extracts were evaluated for anti-a-glucosidase and tyrosinase. The results showed that %inhibition valued for the crude hexane, EtOAc and methanol extracts of anti-a-glucosidase were 98.58 ± 0.09, 98.90 ± 0.21, and 6.68 ± 0.79, respectively. The anti-tyrosinase for the crude hexane, EtOAc and methanol extracts were 97.31 ± 2.77, 97.31 ± 3.34 and 64.94 ± 3.34, respectively. Notably, the bioassay-guided showed that the crude hexane and EtOAc were the highest for anti-a-glucosidase and tyrosinase activities, which led to the interest in purifying the major compound. The isolation and purification were successively done using chromatographic techniques. The pure compounds were determined by spectroscopic analysis (UV, FTIR, and NMR) and comparison with previously reported. The isolation led to three compounds (1-3), ergosterol (1), indole-3-carbaldehyde (2), and 4-hydroxybenzoic acid (3). The molecular docking of compounds 1-3, reported as α-glucosidase and tyrosinase inhibitors, was done by the AutoDockTools 1.5.6 (ADT) software. The results revealed that ergosterol (1) provides the highest binding free energy (-9.91 kcal/mol) to the active site of a-glucosidase, indicating that it is quite specific to this enzyme. Moreover, ergosterol (1) has strong binding energy (-9.64 kcal/mol) to tyrosinase but not significant tyrosinase specificity. This evidence supports that C. Comatus would be applicable for a functional food for controlling glucose blood levels.

Isolation, characterization and antimicrobial activity study of Thymus vulgaris

Herbal medicines are important for ensuring sustainable development goals (SDGs) in healthcare, particularly in developing countries with high rates of antimicrobial resistance (AMR) and little access to medical facilities. Thymus vulgaris is a widely used herbal medicinal plant known for its secondary metabolites and antimicrobial properties. The present study involved a comprehensive examination of the isolation, characterization, and antibacterial activity of Thymus vulgaris obtained from Ethiopia. The aerial part of the plant Thymus vulgaris was successively extracted with hexane, chloroform, and methanol based on differences in polarity. Phytochemical screening tests conducted against hexane, chloroform and MeOH crude extracts indicated the presence of some secondary metabolites. Based on the thin-layer chromatography tests, the chloroform extract was subjected to column chromatography, yielding Tv-2 compounds, namely 5-isopropyl-2-methylphenol. The structures of the compounds were elucidated via spectroscopic methods (UV–Vis, FT-IR and NMR). We investigated the antibacterial properties of hexane crude extract, chloroform crude extract, MeOH crude extract, and isolated fractions derived from T. vulgaris against various bacterial strains. This study contributes to a better understanding of the bioactive components present in Thymus vulgaris crude extracts and their potential role in tackling microbial infections.

Cytotoxic properties, glycolytic effects and high-resolution respirometry mitochondrial activities of Eriocephalus racemosus against MDA-MB 231 triple-negative breast cancer

IntroductionTriple-negative breast cancer (TNBC) represents a significant global health crisis due to its resistance to conventional therapies and lack of specific molecular targets. This study explored the potential of Eriocephalus racemosus (E. racemosus) as an alternative treatment for TNBC. The cytotoxic properties and high-resolution respirometry mitochondrial activities of E. racemosus against the MDA-MB 231 TNBC cell line were evaluated.MethodsHexane solvent and bioactive fraction extractions of E. racemosus were performed, while mass spectrometry-based metabolite profiling was used to identify the phytochemical constituents of the extracts. The extracts were further tested against MDA-MB 231 TNBC cells to determine their cytotoxicity. The mode of cell death was determined using flow cytometry. The activities of caspases 3, 8, and 9 were assessed using a multiplex activity assay kit. Glycolytic activity and High-resolution respirometry measurements of mitochondrial function in the MDA-MB 231 cell line were conducted using the Seahorse XFp and Oroboros O2K.ResultsMetabolite profiling of E. racemosus plant crude extracts identified the presence of coumarins, flavonoids, sesquiterpenoids, triterpenoids, and unknown compounds. The extracts demonstrated promising cytotoxic activities, with a half maximal inhibitory concentration (IC50) of 12.84 µg/mL for the crude hexane extract and 15.49 µg/mL for the bioactive fraction. Further, the crude hexane and bioactive fraction extracts induced apoptosis in the MDA-MB-231 TNBC cells, like the reference drug cisplatin (17.44%, 17.26% and 20.25%, respectively) compared to untreated cells. Caspase 3 activities confirmed the induction of apoptosis in both cisplatin and the plant crude extracts, while caspase 8 and 9 activities confirmed the activation of both the intrinsic and extrinsic apoptosis pathways. Increased levels of glycolytic activity were observed in the hexane crude extract. High-resolution respiratory measurements showed elevated mitochondrial activities in all mitochondrial states except for complex-IV activity.ConclusionThese findings support further exploration of E. racemosus as a potential therapeutic agent for TNBC, offering a promising avenue for the development of targeted treatments with minimal adverse effects.

Effects of Abelmoschus esculentus Extracts and Fractions on Embryos and Adult Individuals of Biomphalaria glabrata (Say, 1818) and on Schistosoma mansoni Cercariae

Schistosomiasis is a neglected tropical disease caused by parasitic worms of the genus Schistosoma. In Brazil, there are reports of infection by the Schistosoma mansoni species, which has the Biomphalaria glabrata snail as one of its intermediate hosts. The present work aimed to test the effects of different Abelmoschus esculentus seed extracts and fractions on adults and embryos of B. glabrata and S. mansoni cercariae. A total of four crude extracts and thirteen fractions with different organic solvents were used for the bioassays. The extracts were analyzed using high-performance liquid chromatography coupled to mass spectrometry. Molluscicidal activity was assessed in 24-well plates, after which the LC50 and LC90 were calculated. Assays with B. glabrata embryos and S. mansoni cercariae were also performed. These findings indicate the presence of flavanoids in the hexane, ethyl acetate, and ethanol crude extracts. For the molluscicidal activity assays, eight fractions had an LC90 value less than that recommended by the WHO. The methanol fraction of the dichloromethane extract (FrMeOH EDM) had the most promising results, with an LC90 of 37.15 mg/L and 100% mortality in embryos of B. glabrata and cercariae. FrMeOH, EDM, and other fractions are possible candidates for new drugs for the combat of schistosomiasis.

Anti-Inflammatory Effects of Moringa Oleifera Lam Leaf Extract in Lipopolysaccharide-Activated Microglia

Moringa oleifera Lam has been used as a medicinal plant in many Asian countries including Thailand. The plant has a variety of pharmacological effects such as antioxidant, anti-inflammation, hypolipidemic, anti-infection, anticancer, and antidiabetic. The leaves and young pods, which contain various nutritional compounds including proteins, fatty acids, and vitamins, are widely consumed. In this study, we investigated the effects of hexane, ethyl acetate, methanol and ethanol crude extracts of M. oleifera Lam leaves on the viability of microglial cells and on the lipopolysaccharide-induced microglial activation. which is widely used as a model for neuroinflammation. Our findings suggested that all of the crude extracts at concentrations ranging from 10–9 to 10–5 g/mL did not significantly affect the viability of microglial cells and exhibited similar effects. Subsequently, we focused on ethanol extract for investigation of the molecular mechanism involving neuroinflammation. We found that the ethanol extract of M. oleifera Lam leaves at the concentrations of 10–9 to 10–5 g/mL significantly reduced inducible nitric oxide synthase (iNOS) protein expression and nitric oxide (NO) production, accompanied by the reduction of phospho-NF-κB and phospho-IκBα expressions. The results suggested that the ethanol extract of M. oleifera Lam leaves reduced NO production through reduction of iNOS protein expression following the inhibition of NF-κB/IκBα signaling pathways. Our study underscored the therapeutic potential of M. oleifera Lam leaves extract in neurodegenerative diseases associated with neuroinflammation. HIGHLIGHTS The ethanol extract of M. oleifera Lam leaves dose-dependently reduced NO production in LPS-activated microglia The reduction in NO production correlated with the reduction in iNOS protein expression The extract inhibited the expression of NF-kB/I-kBα protein expression in a dose-dependent manner The ethanol extract of oleifera Lam leaves exhibited anti-inflammatory effect via the inhibition of NF-kB/I-kBα signaling pathway GRAPHICAL ABSTRACT

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.

Chemotaxonomy Significant in Delimitation of Six Taxa of Euphorbia L. in Iraq

Six taxa of Euphorbia L. (E. hirta L., E. milii var. milii, E. milii cv. White, E. prostrata Aiton, E. hypericifolia L. and E. serpens Kunth) collected from different location in Baghdad were extracted and the phytochemical screening of hexane crude extracts of six taxa of Euphorbia revealed that the most distributed phytochemical compounds were saponins in all taxa except E. prostrata, however, alkaloids have been reported in E. hypericifolia, E. milii cv. White and E. milii var. milii, on the other hand, glycosides have been stated in E. prostrata and E. serpens. Flavonoids and phenols have been noticed in E. serpens only, while terpenes and tannins found in E. prostata and E. hirta respectively. However, the Gas Chromatography-Mass Spectrum of methanol extract of six taxa of Euphorbia analysis explained the presence of twenty seven different chemical compounds and two new terpenoids (trans-Geranylgeraniol and Squalene) from E. milii cv. White. Agglomerative Hierarchical cluster analysis (AHC) stated close chemical relations between all the studied species, the first group comprises E. hirta and E. hypericifolia showed close relationship due to the presence of natural compounds as fatty acids, esters and aromatic alcohols However, E. prostata and E. serpens were close correlated in one group together which characterized by presence of phenols, esters and fatty acids. The anthor group contains E. milli var. milli and E. milli cv. White characterized by the abundance of di- and tri-terpenoids in addition to other natural chemical products.

Nutritional and anti-nutritional properties of sweet cassava (Manihot esculenta) and black pepper (Piper nigrum) leaves

The proximate compositions,some minerals,some vitamins and antinutrients contents of sweet Cassava (Manihot esculenta) and Black pepper (Piper nigrum) leaves were investigated on dried weight basis by employing the standard methods described by the Association of Official Analytical Chemists (AOAC, 1990)., Association of vitamin chemists (1987) and Harborne. (1973). The results showed that Sample A (sweet cassava leabes ) contained crude protein (11.70 ± 2.34%), Hexane extract (1.47 ± 0.1%), Ash content (1.90 ±0.1%), crude fibre (3.56 ± 0.14%), moisture (10.8 ± 0.00%) and carbohydrate (70.57 ± 0.45%).while Sample B(black pepper leaves ) contained crude protein (8.73±1.25%), Hexane extract (1.49 ± 0.1222%), Ash content (1.82 ± 0.141%), crude fibre (2.92 ± 0.141%), moisture (12.81 ± 0.00%) and carbohydrate (78.46 ± 1.25%). The results also revealed that Sample A contained potassium (155.75 ± 1.02 mg/100g), sodium (3.82 ± 0.14 mg/100g), calcium (2.47 ± 0.1 mg/100g), magnesium (75.28 ± 0.60 mg/100g), manganese (0.44 ± 0.10 mg/100g) and copper (0.17 ± 0.01 mg/100 g). Sample B on the other hand, contained potassium (164.45 ± 0.387 mg/100 g), sodium (4.80 ± 0.01 mg/100 g), calcium JJ(3.65 ± 0.223 mg/100 g), magnesium (94.75 ± 0.223 mg/100 g), manganese (0.10 ± 0.001 mg/100 g) and copper (0.17 ± 0.01 mg/100 g). The results of vitamin analysis depicted that Sample A contained vitamin A (0.116 ± 0.01IU), vitamin B1 (0.032 ± 0 mg/100 g), vitamin B2 (0.0027 ± 0 mg/100 g), vitamin B6 (0.125 ± 0 mg/100 g), vitamin B12 (0.024 ± 0 mg/100 g), vitamin C (17.80 ± 0 mg/100 g), vitamin E (0.012 ± 0 mg/100 g) and vitamin K (0.002 ± 0 mg/100 g). While Sample B contained vitamin A (0.114 ± 0.01 IU), vitamin B1 (0.0765 ± 0.038 mg/100 g), vitamin B2 (0.0435 ± 0.038 mg/100 g), vitamin B6 (0.1245 ± 0.022 mg/100 g), vitamin B12 (0.033 ± 0.031 mg/100 g), vitamin C (15.72 ± 0.282 mg/100 g), vitamin E (0.014 ± 0 mg/100 g) and vitamin K (0.00165 ± 0.036 mg/100 g).The results of antinutrient analysis also depicted that Sample A contained Tannin (0.3655±0.038 mg/100 g), saponin (0.2855 ± 0.0387 mg/100 g), Alkaloid (0.1025 ± 0.1025 mg/100 g), phytate (0.2815 ± 0.0387 mg/100 g), oxtate (0.195 ± 0.0316 mg/100 g), glycocyanide (0.005 ± 0.001 mg/100 g) and flavnoid (0.58 ± 0.04 mg/100 g). However, Sample B contained Tannin (0.357 ± 0.031 mg/100 g), saponin (0.293 ± 0.031 mg/100 g), Alkaloid (4.551 ± 0.114 mg/100 g), pytate (0.347 ± 0.031 mg/100 g), oxtate (0.233 ± 0.0446 mg/100 g), glycocyanide (0.005 ± 0.001 mg/100 g) and flavonoid (0.6425 ± 0.05 mg/100 g). Therefore, dried sweet cassava and black pepper leaves are good soourses of proteins, carbohydrates, fibre, potassium, vitamin C and flavonoids. Black pepper leaves also rich in alkaloid.

Cytotoxicity Activity, Metabolite Profiling, and Isolation Compound from Crude Hexane Extract of Cleome rutidospermae.

This study isolated the chemical compounds and evaluated the cytotoxic activity of the crude hexane extract of Cleome rutidospermae herb (CRH). The isolate was purified using silica gel, column chromatography, and preparative thin layer chromatography (PTLC). Furthermore, the structure of the compounds was identified by spectroscopic methods using 1D, 2D NMR, and mass spectrometry. The cytotoxic activity of CRH at a concentration of 20 ug/mL was also tested against MCF-7, A549, KB, KB-VIN, and MDA-MB-231 cancer cells using the sulforhodamine B (SRB) method. The CRH contained compounds of unsaturated fatty acid, saturated fatty acid, lipid, glycerol, ω-3 fatty acid, and cholesterol. Two compounds were obtained from the plant, and their structures were identified as (1) Stigmasta-5,22-dien-3-ol (STML) and (2) 1,2-Benzene dicarboxylic acid, 1,2-bis (2-Ethylhexyl) esters (DEHP). These compounds were reported in this plant for the first time. In comparison, CRH had % growth inhibition in the proliferation of MCF-7 cells up to 28.1%, with cancer cells A549, KB, KB-VIN, and MDA-MB-231 by >50% Compared to the negative DMSO of 0.20%, while the positive control could inhibit the growth of all cancer cells (100%). Our findings suggested that crude herb from the plant CRH was the potential for breast cancer treatment.

Evaluation of The Extract of Microalgae and Its Fatty Acids on Candida spp. Isolated from Renal Impairment Patients

Background: Due to harmful microorganisms’ resistance to many antibiotics, alternative solutions are required to eradicate them. Method: Chlorella vulgaris and Oscillatoria limnetica, two species of microalgae, were isolated for this investigation from the surrounding aquatic environment. The hexane extract of these two algae was tested in vitro by using two activities: antifungal and antibiofilm, against two species of Candida (Candida albicans and Candida glabrata), which were clinically isolated from 100 renal impairment patients, and it was diagnosed by some biochemical test. Results: Fourier Transform Infrared Spectroscopy analysis determined 15 clear bands from (3381.21- 671.23 cm-1) for Chlorella vulgaris, and 12 clear bands from (2960.73- 1261.45 cm-1) for Oscillatoria limnetica. The Gas chromatography-mass spectrometry showed important chemical compounds for both microalgae which: p-Xylene, Dihydrocoumarin, 4,4,5,7,8-pentamethyl, Butylated Hydroxytoluene, 1,4-Benzenediol, 2,5-bis (1,1-dimethylethyl), Eicosane, n-Hexadecanoic acid, 9-Octadecenoic acid, (E), 2-hydroxy-1-(hydroxymethyl) ethyl ester, Bis(2-ethylhexyl) phthalate, 2,3-dihydroxypropyl ester, diethylhydroxylamine, Ethylbenzene, Benzaldehyde, 9,12,15- Octadecatrienoic acid, methyl ester, Hexadecanoic acid, Phytol, Methyl stearate. The results showed that crude hexane extract of Chlorella vulgaris was better than Oscillatoria limnetica as an antifungal, and the percent of antibiofilm was 79% with Chlorella vulgaris, while 77.7% with Oscillatoria limnetica. The rich fatty acids in Chlorella vulgaris have been partially purified from it and applied with two concentrations (10 and 100 mg/ml) as a good antifungal and antibiofilm with a percent of 81.667% for the two concentrations. Conclusion: With the global rise in antibiotic resistance, these algae have demonstrated their value as projects for alternative medicines

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.

Evaluating biological ethnopharmaceutical activities of fruit and root extracts of Solanum incanum L.

Solanum incanum L. is a plant that belongs to the Solanaceae family, growing in tropical climates all over the world. Despite the extensive use of the root and fruit sap extract of S. incanum in the agricultural and ethnopharmaceutical industries in Africa, there is minimal scientific documentation regarding its pesticide bioactive phytochemicals and efficacy. The study investigated the photochemical content (phenolics, flavonoids, saponins, terpenoids and alkaloids) of the methanol, ethyl acetate and hexane crude extracts of the root and fruit of S. incanum. The potential radical scavenging, (antioxidant), cytotoxicity, and efficacy activities of all crude extracts were examined using the 1, 1-diphenyl-2picrylhydrazyl (DPPH), propidium iodide (PI) and aphid leaf dip (ALD) assays respectively. The total phenolic, alkaloid, flavonoid, terpenoid and saponin content were highest in the methanol fruit (MFE), (3.49 ± 1.16 mg GAE/100 mg), steroidal fruit (SAE), (0.14±0.06 mg ATP/100 mg, hexane fruit (HFE), 3.62±1.23 mg QCT/100 mg, hexane root (Hroot), (1.19±0.47 mg LIN/100 mg and saponin (3.74±1.17 mg/100 mg). The aqueous extract of the ethyl acetate fruit had the highest antioxidant activity with an IC50 of 35.53±0.53 μg·mL-1 (p < 0.05) The methanol fruit and steroidal alkaloid extracts exhibited potent cytotoxicity against Vero cells demonstrating the highest cell mortality rate of 98% and 70% respectively at 100 μg·mL-1. Melphalan which was used as a positive control had a mortality rate of 75% cell at 100 μg/mL concentration. The methanol fruit extract exhibited the highest efficacy against the cabbage aphids with an LD50 of 13.16±3.78 μg·mL-1 (p < 0.05). The results indicated that the fruit and root of S. incanum have a variety of phytochemicals that may be considered for use as antioxidants and/or pesticides. By exhibiting the most potent cytotoxic activity against Vero cells, S. incanum roots have the greatest potential for the discovery of cytotoxic compounds necessary to develop anticancer drugs. The extracts with high potent efficacy against cabbage aphids need to be investigated further as pesticides to benefit the disadvantaged communities in the Southern African region. Key words: Antioxidant, cytotoxicity, lethal dose, mortality rate, phytochemicals, quantitative analysis

Antiparasitic properties of 4-nerolidylcatechol from Pothomorphe umbellata (L.) Miq. (Piperaceae) in vitro and in mice models with either prepatent or patent Schistosoma mansoni infections

Ethnopharmacological relevanceRoots of Pothomorphe umbellata (L.) Miq. are used in traditional medicine of Africa and South America for the treatment of malaria and helminthiasis. However, neither P. umbellata nor its isolated compounds have been evaluated against Schistosoma species. Aims of this studyTo investigate the antischistosomal effects of P. umbellata root extracts and the isolated compound 4-nerolidylcatechol (4-NC) against Schistosoma mansoni ex vivo and in murine models of schistosomiasis. Materials and methodsThe crude hydroalcoholic (PuE) and hexane (PuH) extracts of P. umbellata roots were prepared and initially submitted to an ex vivo phenotypic screening against adult S. mansoni. PuH was analyzed by HPLC-DAD, characterized by UHPLC-HRMS/MS, and submitted to chromatographic fractionation, leading to the isolation of 4-NC. The anthelmintic properties of 4-NC were assayed ex vivo against adult schistosomes and in murine models of schistosomiasis for both patent and prepatent S. mansoni infections. Praziquantel (PZQ) was used as a reference compound. ResultsPuE (EC50: 18.7 μg/mL) and PuH (EC50: 9.2 μg/mL) kill adult schistosomes ex vivo. The UHPLC–HRMS/MS analysis of PuH, the most active extract, revealed the presence of 4-NC, peltatol A, and peltatol B or C. After isolation from PuH, 4-NC presented remarkable in vitro schistosomicidal activity with EC50 of 2.9 μM (0.91 μg/mL) and a selectivity index higher than 68 against Vero mammalian cells, without affecting viability of nematode Caenorhabditis elegans. In patent S. mansoni infection, the oral treatment with 4-NC decreased worm burden and egg production in 52.1% and 52.3%, respectively, also reducing splenomegaly and hepatomegaly. 4-NC, unlike PZQ, showed in vivo efficacy against juvenile S. mansoni, decreasing worm burden in 52.4%. ConclusionsThis study demonstrates that P. umbellata roots possess antischistosomal activity, giving support for the medicinal use of this plant against parasites. 4-NC was identified from P. umbellata roots as one of the effective in vitro and in vivo antischistosomal compound and as a potential lead for the development of novel anthelmintics.

Prospects for Pharmaceutical of Zingiber officinale Extracts and Fractions by Analysis of the Chemical Markers, Safety Profile, Anti-Lipoxygenase, and Analgesic Activity

Ginger (Zingiber officinale) has gained recognition for its flavoring uses and its numerous health benefits. Ginger can be used in functional foods and nutraceuticals alone or combined with other herbal plants. This study aims to validate the pharmaceutical potential of the Zingiber officinale variety from the Philippines by examining the chemical composition, acute oral toxicity, anti lipoxygenase, and analgesic activities of its extracts and fractions. 6, 8, 10-Gingerols and 6, 8, 10-Shogaols were identified by high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) in methanol and hexane extracts. Acute oral toxicity in mice indicated the median lethal dose (LD50) of methanol and hexane extracts of Zingiber officinale, and the 1% IPA-hexane and 50% IPA-hexane fractions obtained by solid phase extraction (SPE) was greater than 2000 mg/kg body weight. The 1% IPA-hexane fraction altered the behavioral pattern of the animals. The anti-inflammatory activity was analyzed by lipoxygenase (LOX) inhibition assay. The IC50 value for the inhibitory action of crude methanol extract on LOX was 218.00 µg/mL. The crude hexane extract IC50 value was 57.40 µg/mL. SPE fractionation of hexane extracts resulted in higher IC50 values; 72.14 µg/mL for the 1% IPAhexane fraction and 98.65 µg/mL for the 50% IPA-hexane fraction. Through in-vitro suppression of LOX, the hexane crude extracts showed more effective anti-inflammatory properties. The analgesic effect was measured using acetic acid-induced writhing in mice. All doses of the methanol and hexane extract inhibited writhing by a percentage ranging from 91-100%. The SPE fractions of the hexane extract have 78-99% inhibition. Both extracts and fractions demonstrated strong analgesic effects in the mice models.

Chemical Composition, Anti-Inflammatory and Analgesic Activities of Extracts and Fractions of Vitex negundo

This study aims to determine chemical components and demonstrate the anti-inflammatory and analgesic activities of Vitex negundo species native to the Philippines. Using high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), eleven compounds were tentatively identified from the iridoids, flavonoids, and phenolics group. The anti-inflammatory activity was determined by measuring the inhibition of proinflammatory enzymes, lipoxygenase (LOX), and cyclooxygenase (COX-2). For LOX inhibitory effect, crude methanol extract has an IC50 value of 440.9 µg/mL. Solid-phase extraction (SPE) fractionation results in IC50 values of 196.6 µg/mL and 150.0 µg/mL for the 50% methanol-water and 100% methanol fractions, respectively. The IC50 value of crude hexane extract is 519.7 µg/mL. SPE produced lower IC50 values; 397.5 µg/mL for 2% IPA-hexane fraction and 142.9 µg/mL for 50% IPA-hexane fraction. The inhibition against COX-2 was effective in the 100% methanol fraction with an IC50 of 34.27 μg/mL and 2% IPA-hexane with an IC50 of 6.57 μg/mL. SPE fractions demonstrated more potent anti-inflammatory effects via in-vitro inhibition of LOX for the 100% methanol-water and 50% IPA-hexane or COX-2 for the 100% methanol and 2% IPA-hexane. The analgesic action was evaluated using acetic acid-induced writhing in mice. The percentage inhibition of writhing at all dose levels of methanol and hexane extracts ranged from 90-96% (p<0.05). The SPE fractions from methanol extracts showed inhibition of 73-97% (p<0.05). Both extracts and fractions showed strong analgesic effects in an in-vivo animal model.

Chemical Constituents and Their Anti-inflammatory Activities from the Stems of Cryptolepis buchanani

The stems of Cryptolepis buchanani are traditionally used for the treatment of inflammation, including arthritis, muscle and joint pain. In the present study, 3 crude extracts (hexane, EtOAc and methanol) from the stems of C. buchanani were evaluated for anti-inflammatory effects using an in vitro model of RAW 264.7 macrophages. At a concentration of 50 μg/mL, the hexane and EtOAc crude extract have inhibited nitric oxide (NO) production by 84.33±2.01 and 82.49±0.92 %, respectively. 3,4-Dihydroxyl benzoic acid (1), vanillic acid (2), syringaldehyde (3), isoscopoletin (4) and stigmast-4-ene-3,6-dione (5) were isolated from hexane and EtOAc crude extracts. Their structures were identified on the basis of 1H and 13C NMR, as well as a comparison of the data from previous reports. Compounds 1-5 were evaluated for anti-inflammatory activity. Compound 3 showed activity of NO inhibition with IC50 values of 49.07 μM (Indomethacin IC50 values of 39.21 μM). In addition, compounds 1-3 and 5 were isolated for the first time from this plant. HIGHLIGHTS The crude hexane and ethyl acetate (EtOAc) (each, 50 mg/mL) inhibited nitric oxide (NO) production with inhibition percentage at 84.33 and 82.49, respectively 3,4-Dihydroxyl benzoic acid, vanillic acid, syringaldehyde and stigmast-4-ene-3,6-dione were isolated for the first time from this plant Syringaldehyde showed activity of NO inhibition with IC50 values of 49.07 μM (Indomethacin IC50 values of 39.21 μM) GRAPHICAL ABSTRACT

Cytotoxic activity of Cape Fynbos against triple-negative breast cancer cell line

• Erica glabella, Hippia frutescens and Salvia africana-lutea ( methanol), and Eriocephalus racemosa ( hexane) extracts IC 50 at < 30 µg/mL • Eriocephalus racemosa induced 21.48±2.86% apoptosis similar to cisplatin 23.72±4.36%. • Metabolite profiling of Salvia africana-lutea extract revealed the presence of phosphatidylcholines, triterpenoids, oxepane and 7-O-methylated flavonoids derivatives. Breast cancer is a leading cause of cancer-related deaths in women globally. Even though a plethora of treatments are available, most patients experience adverse effects which affect their quality of life. The aim of this study was to investigate the utility of nine Fynbos plants ( Erica magnisylvae E.G.H Oliv., Erica canescens J.C. Wendl. , Erica coccinea L. , Erica glabella Thunb. , Erica corifolia L. , Eriocephalus racemosa L. , Hippia frutescens L., Salvia africana-lutea L. and an unknown Fynbos plant) in treating breast cancer. Solvent extraction of fynbos plants was performed, and crude extracts were evaluated against MDA-MB 231 cancer cell line to determine the cytotoxic activity with the mode of cell death confirmed using flow cytometry. Antioxidant activity and mass spectrometry-based metabolite profiling were performed to characterize and identify the phytochemical constituents of the extracts. The methanol extracts from E. glabella ., H. frutescens and S. africana-lutea and the hexane extract from E. racemosa showed promising cytotoxic activities in the screening phase and thus were further evaluated to determine their 50% inhibitory concentrations (IC 50 ) which were found to be < 30 µg/mL. Flow cytometry analysis of treated MDA-MB 231 cells revealed promising results for the hexane crude extract (leaves) of E. racemosa and stem methanol crude extract of H. frutescens which induced apoptosis in MDA-MB-231 cancer cell line, similar to the reference drug cisplatin. Metabolite profiling of S. africana-lutea extract, the most potent apoptosis inducer in this study, revealed the presence of phosphatidylcholines, triterpenoids, oxepane and 7-O-methylated flavonoids derivatives. It was concluded that E. racemosa and S. africana-lutea are excellent candidates for further development of therapeutic agents in the fight against cancer, given the pressing need for novel efficacious agents.

Phytochemical profiling, antioxidant activity and antimicrobial activity of Argyrcia speciosa (Linn.f.) Sweet., collected from Darrang district, Assam

The present study was carried out to analyze the phytochemical constituents, antioxidant activity, totalphenolic content, total flavonoid content, and antimicrobial activity of Argyrcia speciosa (Linn.f.) Sweet.Preliminary phytochemical analysis was done using standard procedure. Total phenolic content, totalflavonoid content, and antioxidant activities were determined spectrophotometrically in crude aqueous,methanol, chloroform, and hexane extracts. The in vitro antimicrobial activity was determined by well. diffusion method against seven MTCC bacterial species and two MTCC fungal species. Preliminaryphytochemical analysis indicated the presence of saponin, tannin, flavonoid, phenol, trapezoid, cardiacglycoside, and alkaloid, except anthraquinone, steroid, and reducing sugar. The chloroform extract showeda better result than the other extract. Clear antimicrobial activity was observed against Staphylococcusepidermidis and Escherichia coli using methanolic extract while aqueous extract showed moderate activity.Moderate activity was also noted against S. epidermidis and P. vulgaris using methanol and chloroformextract respectively. The percentages inhibition in the case of DPPH and ABTS were recorded highest as83.15 ± 0.000% and 94.80 ± 0.000 for chloroform and methanol extract respectively. The present studyrevealed that both primary and secondary metabolites are present in Argyrcia speciosa (Linn. f.) Sweet., which confer their traditional uses as food and medicine.

Insecticidal effects of some selected plant extracts against Anopheles stephensi (Culicidae: Diptera)

BackgroundThe use of synthetic insecticides against mosquitoes may lead to resistance development and potential health hazards in humans and the environment. Consequently, a paradigm needs to shift towards the alternative use of botanical insecticides that could strengthen an insecticide resistance management programme. This study aimed to assess the insecticidal effects aqueous, hexane, and methanol crude leaf extracts of Calpurnia aurea, Momordica foetida, and Zehneria scabra on an insectary colony of Anopheles stephensi larvae and adults.MethodsFresh leaves of C. aurea, M. foetida and Z. scabra were collected and dried, then separately ground to powder. Powdered leaves of test plants were extracted using sonication with aqueous, hexane, and methanol solvents. The extracts were concentrated, and a stock solution was prepared. For comparison, Temephos (Abate®) and control solutions (a mixture of water and emulsifier) were used as the positive and negative controls, respectively. Different test concentrations for the larvae and the adults were prepared and tested according to WHO (2005) and CDC (2010) guidelines to determine lethal concentration (LC) values. Mortality was observed after 24 h exposure. The statistical analyses were performed using Statistical Package for the Social Sciences (SPSS) software (Kruskal-Wallis test) and R software (a generalized linear model was used to determine LC50 and LC90 values of the extracts).ResultsThe lowest LC50 values were observed in aqueous extracts of M. foetida followed by Z. scabra extract and C. aurea leaves at 34.61, 35.85, and 38.69 ppm, respectively, against the larvae. Larval mortality was not observed from the hexane extracts and negative control, while the standard larvicide (temephos) achieved 100% mortality. Further, the adulticidal efficacy was greatest for aqueous extract of Z. scabra with LC50 = 176.20 ppm followed by aqueous extract of C. aurea (LC50 = 297.75 ppm).ConclusionThe results suggest that the leaf extracts of the three test plants have the potential of being used for the control of vector An. stephensi larvae and adult instead of synthetic mosquitocides. Further studies need to be conducted to identify the active ingredients and their mode of action.