Hexane Fraction Research Articles

α-Glucosidase, Angiotensin-converting Enzyme, and Acetylcholinesterase Inhibitory Activities of a Marine Red Alga Galaxaura oblongata

Seaweeds have gained interest from the pharmaceutical industry due to their diverse secondary metabolites with potential applications in the prevention and treatment of lifestyle diseases. In spite of the abundance of seaweeds in the coastal area of Iligan Bay in the Philippines, there has been limited investigation into their pharmacological properties. Therefore, this study aimed to evaluate the in vitro α-glucosidase inhibition, angiotensin-converting enzyme (ACE) inhibition, and acetylcholinesterase (AChE) inhibition properties of various fractions obtained from the marine red alga Galaxaura oblongata. The methanol extract of G. oblongata was sequentially partitioned into hexane, dichloromethane, ethyl acetate, butanol, and methanol fractions. All assays were performed in vitro and microplate based. The results of α-glucosidase inhibition activity assay showed that the n-hexane fraction and dichloromethane fraction exhibited greater than 50% inhibitory activity at 200 ppm. Furthermore, ACE inhibition activity assay revealed that dichloromethane fraction, ethyl acetate fraction, and butanol fraction displayed ACE inhibition activity above 50% at 250 ppm. In addition, the hexane fraction, dichloromethane fraction, and ethyl acetate fraction demonstrated potent AChE inhibitory activity at 250 ppm. Overall, the findings show that G. oblongata has antidiabetic, anti-hypertensive, and neuroprotective potentials. This is the first report of the in vitro α-Glucosidase, ACE, and AChE inhibition activities of G. oblongata. Further investigation and purification of the highly potent fractions in each assay is highly suggested to identify and characterize the compounds responsible for the observed bioactivities, which could serve as possible leads for drug discovery efforts in the management of various lifestyle diseases.

Modulation of quorum sensing and biofilm of Gram-negative bacterial pathogens by Cinnamomum zeylanicum L.

The development of antibiotic resistant microbial pathogens has become a global health threat and a major concern in modern medicine. The problem of antimicrobial resistance (AMR) has majorly arisen due to sub-judicious use of antibiotics in health care and livestock industry. A slow progress has been made in last two decades in discovery of new antibiotics. A new strategy in combatting AMR is to modulate or disarm the microbes for their virulence and pathogenicity. Plants are considered as promising source for new drugs against AMR pathogens. In this study, fraction-based screening of the Cinnamomum zeylanicum extract was performed followed by detailed investigation of antiquorum sensing and antibiofilm activities of the most active fraction that is, C. zeylanicum hexane fraction (CZHF). More than 75% reduction in violacein pigment of C. violaceum 12472 was overserved. CZHF successfully modulated the virulence of Pseudomonas aeruginosa PAO1 by 60.46%-78.35%. A similar effect was recorded against Serratia marcescens MTCC 97. A broad-spectrum inhibition of biofilm development was found in presence of sub-MICs of CZHF. The colonization of bacteria onto the glass coverslips was remarkably reduced apart from the reduction in exopolymeric substances. Alkaloids and terpenoids were found in CZHF. GC/MS analysis revealed the presence of cinnamaldehyde dimethyl acetal, 2-propenal, coumarin, and α-copaene as major phytocompounds. This study provides enough evidence to support potency of C. zeylanicum extract in targeting the virulence of Gram -ve pathogenic bacteria. The plant extract or active compounds can be developed as successful drugs after careful in vivo examination to target microbial infections. RESEARCH HIGHLIGHTS: Hexane fraction of Cinnamomum zeylanicum is active against QS and biofilms. The broad-spectrum antibiofilm activity was further confirmed by microscopic analysis. Dimethyl acetal, 2-propenal, coumarin, α-copaene, and so forth are major phytocompounds.

HPLC and GC Characterization of Dicliptera bupleuroides Aerial Parts and Evaluation of Its Anti-Inflammatory Potential in Vitro, in silico and in Vivo Using Carrageenan and Formalin Induced Inflammation in Rat Models.

The current study aimed to evaluate the anti-inflammatory activity of Dicliptera bupleuroides Nees aerial parts methanol extract and its different fractions namely hexane, chloroform, ethyl acetate and butanol in vitro using cyclooxygenase inhibitory assay (COX-2). In vivo anti-inflammatory evaluation was performed using carrageenan and formalin induced inflammation in rat models followed by molecular docking. High performance liquid chromatography (HPLC) and gas chromatography coupled with mass chromatography (GC/MS) analyses were used for chemical analyses of the tested samples. The tested samples showed significant inhibition in COX-2 inhibitory assay where methanol extract (DBM) showed the highest inhibitory potential at 100 μg/mL estimated by 67.86 %. At a dose of 400 mg/kg, all of the examined samples showed pronounced results in carrageenan induced acute inflammation in rat model at 4th h interval with DBM showed the highest efficiency displaying 65.32 % inhibition as compared to the untreated rats. Formalin model was employed for seven days and DBM exhibited 65.33 % and 69.39 % inhibition at 200 and 400 mg/kg, respectively approaching that of the standard on the 7th day. HPLC revealed the presence of caffeic acid, gallic acid and sinapic acid, quercetin and myricetin in DBM. GC/MS analysis of its hexane fraction revealed the presence of 16 compounds belonging mainly to fatty acids and sterols that account for 85.26 % of the total detected compounds. Molecular docking showed that hexadecanoic acid followed by decanedioic acid and isopropyl myristate showed the best fitting within cyclooxygenase-II (COX-II) while nonacosane followed by hexatriacontane and isopropyl myristate revealed the most pronounced fitting within the 5-lipoxygenase (5-LOX) active sites. Absorption, metabolism, distribution and excretion and toxicity prediction (ADMET/ TOPKAT) concluded that most of the detected compounds showed reasonable pharmacokinetic, pharmacodynamic and toxicity properties that could be further modified to be more suitable for incorporation in pharmaceutical dosage forms combating inflammation and its undesirable consequences.

Evaluation of the Anti-Diabetic Activity of Paliasa Leaf Ethanol Fraction (Kleinhovia hospita L) Through Inhibition of the Enzymes $\alpha$-Glucosidase and $\alpha$-Amylase

Kleinhovia hospita has long been used to treat diseases, including diabetes mellitus. This study aimed to evaluate if the hypoglycemic effects of K. hospita extract and fractions were related to its inhibition on the activity of $\alpha$-amylase and $\alpha$-glycosidase enzymes. K. hospita leaves were extracted with 96% ethanol and then subjected to liquid-liquid partition using n-hexane, ethyl acetate, and water as solvents. The inhibitory capacity of ethanolic extract and its fractions were tested against $\alpha$-amylase and $\alpha$-glycosidase enzyme activity and compared to acarbose. The in vitro study revealed that the IC50 of ethanolic extract, hexane, ethyl acetate, and water fractions were 1496,8 µg/mL, 1266,7 µg/mL, 12577,5 µg/mL, and 5217,0 µg/mL, respectively, against alfa-amylase activity. Whereas, for $\alpha$-glycosidase activity, the IC50 values were 1341,2 µg/mL, 25470,0 µg/mL, 64450,0 µg/mL, and 1291,4 µg/mL, respectively. These values were markedly greater than those of the acarbose (7,4 and 5,5 µg/mL, respectively). From the research results, it was concluded that K. hospita leaf extract had a high IC50 value from K. hospita extracts and fractions, which indicated that the hypoglycemic effect of K. hospita was not related to the inhibition of the $\alpha$-amylase and $\alpha$-glycosidase enzymes.

HPLC Analysis and In Vitro and In Silico Evaluation of the Biological Activity of Polyphenolic Components Separated with Solvents of Various Polarities from Helichrysum italicum.

Helichrysum italicum has piqued the interest of many researchers in recent years, mostly for its essential oil, but increasingly for its polyphenolic content as well. In the current study, we examine the polyphenolic composition of H. italicum grown in Bulgaria. The polyphenolic complex was fractionated with solvents of various polarities, including hexane, chloroform, ethyl acetate, and butanol, in order to assess the biological impact of the components. HPLC-PDA and UHPLC-MS/MS were used to examine all fractions. The green coffee fingerprint profile was employed as a "surrogate standard" in the polyphenolic components detection approach. From the UHPLC-MS/MS analysis, we identified 60 components of the polyphenolic complex such as quercetin 3-O-glucuronide, quercetin acetyl-glycoside, isorhamnetin acetyl-glycoside, isorhamnetin caffeoyl-glycoside, quercetin caffeoyl-malonyl-glycoside, isorhamnetin coumaroyl-glycoside, coumaroyl-caffeoylquinic acid, and diCQA-acetyl-derivative were first reported in the composition of H. italicum. The biological activity of the fractions was evaluated in vitro and in silico, which included the fight against oxidative stress (hydrogen peroxide scavenging activity (HPSA), hydroxyl radical scavenging activity (HRSA), metal-chelating activity (MChA)) and nitrosative (nitric oxide scavenging activity) (NOSA)), in vitro anti-inflammatory, and anti-arthritic activity. Results are presented as IC50 ± SD μg/mL. The analysis showed that the EtOAc fraction was characterized by highest HPSA (57.12 ± 1.14 μg/mL), HRSA (92.23 ± 1.10 μg/mL), MChA (5.60 ± 0.17 μg/mL), and NOSA (89.81 ± 2.09 μg/mL), while the hexane and chloroform fractions showed significantly higher in vitro anti-inflammatory activity (30.48 ± 2.33 μg/mL, 62.50 ± 1.69 μg/mL) compared to the standard ibuprofen. All three fractions showed potential anti-arthritic activity (102.93 ± 8.62 μg/mL, 108.92 ± 4.42 μg/mL, 84.19 ± 3.89 μg/mL).

Evaluation of in vitro antiplasmodial, antiproliferative activities, and in vivo oral acute toxicity of Spathodea campanulata flowers

Spathodea campanulata is used in traditional medicine to treat various ailments such as malaria, human immunodeficiency virus (HIV), cancer, fever and urethral inflammation. The aim of this study was to investigate the antiplasmodial, and antiproliferative activities of the extract and resulted fractions from S. campanulata flowers, as well as assessing the acute toxicity of its aqueous fraction. The in vitro cell-growth inhibition activities were assessed against Plasmodium falciparum strain 3D7 for antimalarial activity and three cancer cell lines: Hs683 (human oligodendroglioma), MCF7 (human breast carcinoma), and murine B16F10 (mouse melanoma) for antiproliferative activity while the in vivo acute oral toxicity was determined according to the modified organisation for Economic Co-operation and Development (OECD) guidelines 423 at a fixed dose on Female Wistar strain laboratory rats. The dichloromethane, ethyl acetate and hexane fractions at a concentration of 25 µg/mL each significantly reduced the viability of 3D7 Plasmodium cells with viability percentages of 19.0%, 14.1% and 31.9%, respectively, and IC50 of 28.1, 30.2 and 29.7 µg/mL, respectively. The ethyl acetate fraction showed a moderate antiproliferative activity on mouse melanoma with an I50 value of 54.6 µg/mL. Only the dichloromethane fraction was able to inhibit the 3 cell lines tested with IC50 values less than 15 µg/mL. An oral administration of the aqueous fraction did not induce an abnormal variation of the physiological parameters in female Wistar laboratory rats, at non-toxic doses up to 5000 mg/kg body weight for 14 days. These results confirm the use of this plant in traditional medicine for its antimalarial and anticancer potential.

In vitro antioxidant, antibacterial, cytotoxic, and epigenetic screening of crude extract and fractions of the marine sponge Neopetrosia exigua from Mauritius waters

The marine sponge Neopetrosia exigua is known as a goldmine of novel compounds, yet its pharmacological activities remain poorly characterised. Herein, this study investigates the bioactivities of N. exigua collected from Mauritius waters. The crude extract (dichloromethane: methanol), hexane, ethyl acetate and aqueous fractions obtained from N. exigua were subjected to in vitro antioxidant assays. Their antibacterial activities were evaluated using the broth microdilution method to determine the minimum inhibitory concentration (MIC). The cytotoxic and epigenetic activities were further screened using the MTT assay and a cell-based image system that measures de-repression of a silenced Green Fluorescent Protein (GFP) reporter gene, respectively. Higher antioxidant activity was recorded for the ethyl acetate fraction as demonstrated by its significant ferric reducing antioxidant power, radical scavenging, and metal chelating activities relative to control (p < 0.05). The best antibacterial profile was presented by the ethyl acetate fraction against Cutibacterium acnes (MIC: 0.039 mg/ml), Streptococcus mutans (MIC: 0.078 mg/ml) and Mycobacterium smegmatis (MIC: 0.313 mg/ml). Similarly, the fraction displayed significant cytotoxicity against the human liposarcoma SW872 cells with IC50 value of 44.34 ± 2.64 µg/ml and GFP re-activation capacity of 43.79 ± 3.19% (p < 0.05). This work conveys interesting data on the antioxidant, antimicrobial, and anticancer properties of N. exigua. In particular, this study indicates the promising potential of N. exigua as a reservoir of epigenetically active agents that can modulate transcription of silenced genes involved in carcinogenesis. Hence, further investigations to isolate the active constituents is actively warranted.

Potent anti-cancer activity of Sphaerocoryne affinis fruit against cervical cancer HeLa cells via inhibition of cell proliferation and induction of apoptosis

BackgroundCervical cancer remains a significant global health issue, highlighting the need for effective therapeutic strategies. Given that Sphaerocoryne affinis (SA) has shown potential anti-cancer activity in several cancer types, herein, we investigate the effects of SA fruit (SAF) on human cervical cancer HeLa cells and their underlying mechanisms of action.MethodsSAF extract cytotoxicity was assessed in various cancer cell lines. The effects of the hexane fraction (SAF-Hex) on HeLa cell viability, cell cycle protein expression, apoptosis, and DNA damage were evaluated using cytotoxicity assays, Western blotting, quantitative PCR, 4′,6-diamidino-2-phenylindole (DAPI) staining, and a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.ResultsSAF-Hex selectively inhibited HeLa cell viability with an IC50 of 4.20 ± 0.36 µg/mL and a selectivity index of 5.11 ± 0.58. The time-dependent cytotoxicity assay showed decreased cell survival after 48 h of treatment, accompanied by morphological changes and apoptotic bodies in HeLa cells. SAF-Hex also suppressed HeLa cell cycle proteins (Cyclin E, CDK2, and CDK1), reduced PCNA transcription, and diminished AKT and mTOR activation, thus inhibiting cell proliferation. The increased γH2AX expression, DNA fragmentation, and caspases-3 and -9 activation indicated SAF-Hex-induced DNA damage and apoptosis. However, the BAX/BCL-2 ratio remained unchanged, and BAX and BCL2 expression was attenuated.ConclusionSAF-Hex effectively inhibits HeLa cell proliferation and induces DNA damage in that cervical cancer cell line activating apoptosis through the intrinsic pathway. Interestingly, the BAX/BCL-2 ratio remained unchanged while BAX and BCL2 transcription was attenuated. Hence, further research is required to explore this unexpected finding and facilitate the development of novel therapies targeting cervical cancer HeLa cells.

Acaricidal effects of Tagetes minuta L. (Asteraceae) against Tetranychus truncatus Ehara (Prostigmata: Tetranychidae)

ABSTRACT The study evaluated the efficacy of solvent fractions of Mexican marigold, Tagetes minuta against the spider mite, Tetranychus truncatus. The dried and pulverized botanical was subjected to sequential extraction using three solvents viz., hexane, chloroform and water in the increasing order of polarity. Solvent fractions were evaluated for their ovicidal and adulticidal activities against T. truncatus in the laboratory at five different concentrations viz., 0.025, 0.05, 0.1, 0.15 and 0.2 %. Results indicated that only hexane fraction at higher concentrations of 0.15 and 2.0 % exhibited significant ovicidal action, causing 100.00 and 81.33 per cent egg mortality, respectively. However, all the fractions recorded significant adulticidal effect at higher concentrations with hexane fractions at 0.2 and 0.15 % recording significantly higher adult mortality (>90 %), followed by chloroform fraction. The best concentrations of the three solvent fractions identified in the laboratory study were evaluated against T. truncatus on amaranthus, in a pot culture experiment. Fourteen days post treatment, hexane fraction at 0.2 % reduced mite population by 88.78 per cent and was superior to neem oil emulsion at 2% (76.44 % reduction), which was on par with chloroform and aqueous fractions. Results of the study identified Mexican marigold as a potential candidate in mite pest management, which may be further validated through multi-locational field trials.

Evaluation Of Antibacterial Activity In Pelargonium Graveolens Extracts

The antibacterial activity of the total ethanolic extract and fractions of Pelargonium graveolens against Candida albicans, Escherichia coli, Pseudomonas spp, Staphylococcus aureus and Bacillus spp. Methods: To measure the relative antimicrobial activity, the percentage of inhibition was calculated by the plate and well method. Results: It was established that the highest antimicrobial activity was presented by the oily extract without preference for Gram positive or Gram negative, the acetone fraction with a value of 58.53% inhibition. The hexane fraction has activity greater than 100% only against S. aureus. Conclusions: The extracts of P. graveolens present antimicrobial activity against Gram-positive and Gram-negative bacteria and yeasts, especially the oily extract, which presented the highest percentage of inhibition recorded against Escherichia coli, Staphylococcus aureus and Candida albicans, respectively; while for Pseudomonas aeruginosa and Bacillus subtilis, the extract that presented the highest percentage of inhibition was ethyl acetate.

Chemical composition and biological activities of the extracts of Peperomia pellucida fractions

Background: Peperomia pellucida is a medicinal and vegetable plant used worldwide, representing a multi-purpose vegetable with applications in the pharmaceutical, food and cosmetic industries. Objective: This study evaluated the antioxidant, antibacterial, antifungal, anti-inflammatory, and antidiabetic potential of fractional extracts from P. pellucida plant derived from Can Tho City, Vietnam. Methods: Four fractional extracts were prepared using different polarity solvents (hexane, dichloromethane, ethyl acetate) and used to determine the best extract for each biological property. The fractions’ total alkaloid, phenolic, and flavonoid content were observed. The four extracts were evaluated for their potential bioactivities: antioxidant, antibacterial, antifungal, anti-inflammatory, and antidiabetic. Results: Correspond with the hexane, dichloromethane, ethyl acetate, and aqueous fractions, the total content of alkaloids was determined to be 255 ± 23.8; 157 ± 14.0; 219 ± 6.55; 221 ± 6.23 (mg AE/g extract), the total phenolic content was 112 ± 3.34; 141 ± 1.77; 234 ± 29.5; 123 ± 5.04 (mg GAE/g extract), whereas the total content of flavonoids was 423 ± 22.6; 169 ± 6.30; 1839 ± 18.8; 173 ± 22.6 (mg QE/g extract), respectively. The ethyl acetate fraction gave the best efficiency in DPPH, ABTS, iron reduction, and TAC methods (IC50 = 334 ± 2.10 µg/mL; 51.4 ± 0.41 µg/mL; 79.1 ± 0.40 µg/mL; and 83.0 ± 0.17 µg/mL, respectively). Antibacterial activity was investigated on 5 strains of Bacillus cereus, Escherichia coli, Staphylococcus aureus, Salmonella typhimunum, Pseudomonas aeruginosa; the results showed that the extracts were resistant to 5 strains of bacteria, especially best resistant in 2 fractions of ethyl acetate and aqueous. The minimum inhibitory concentration (MIC) value ranged from 0.5 to 32 mg/mL, while the minimum bactericidal concentration (MBC) value ranged from 16 to 64 mg/mL. The best anti-inflammatory activity was ethyl acetate with an IC50 value of 216.7 ± 7.2 µg/mL, close to that of Diclofenac at 205.4 ± 0.5. The antidiabetic activity was investigated based on the ability to inhibit α-amylase and α-glucosidase enzymes. The results showed that the best α-amylase inhibitors were hexane and dichloromethane (IC50 = 208.83 ± 2.41 and 191.60 ± 1.27 µg/mL, respectively), roughly equal to the acarbose (155.68 ± 2.59 µg/mL). The best α-glucosidase inhibitory fraction was ethyl acetate (IC50 of 157.04 ± 0.23 µg/mL), close to that of acarbose (116.45 ± 0.21 µg/mL). Conclusion: Fractional extracts from P. pellucida distributed in gardens of Can Tho City, Vietnam, contain potential antioxidant, antibacterial, antifungal, anti-inflammatory, and antidiabetic bioactive compounds.

In vitro cytotoxic and apoptotic activity of the Mauritian marine sponge Neopetrosia exigua

Marine sponges belonging to the genus Neopetrosia represent a quasi-inexhaustible source of novel cytotoxic compounds. Yet studies delineating their molecular mechanisms of action in cancer cells remain scarce. We investigated the cytotoxic and apoptosis inducing potential of the Mauritian marine sponge Neopetrosia exigua derived crude extract, hexane and ethyl acetate fraction. Their cytotoxic activity was screened against four cancer cell lines and two non-malignant cell lines via the Alamar Blue metabolic assay. The level of intracellular reactive oxygen species (ROS) production, endogenous antioxidant enzyme activity (catalase and superoxide dismutase) and mitochondrial membrane potential were determined. The ability of the active extract to induce apoptosis in cancer cells and modulate the expression levels of apoptotic markers (caspases and polyADP-ribose polymerase (PARP)) was further evaluated via western blot. The ethyl acetate fraction (NEEAF) displayed the highest inhibitory effect with an IC50 of 6.87 μg/mL against the liver hepatocellular carcinoma cell line (HepG2). Mechanistically, NEEAF induced morphological hallmarks characteristic of apoptosis, increased ROS production, decreased catalase and superoxide dismutase activity and mitochondrial membrane depolarisation in a concentration-dependent manner compared to the control (p&lt;0.05). In addition, NEEAF induced the activation of caspase-9, -7, -3 and cleavage of PARP. Overall, this study provides biochemical evidence for oxidative stress-mediated cytotoxicity and apoptosis in HepG2 cells by NEEAF. Further in-depth investigations are needed to isolate the active constituents, which may potentially lead to the development of novel anticancer therapeutics. Significance: Marine sponges represent an untapped goldmine of structurally unique compounds with interesting anticancer properties. This important initial investigative work will set the stage for more in-depth mechanistic studies and chemical characterisation of potentially novel bioactive compounds from the genus Neopetrosia. This work will also help to strengthen frameworks oriented towards the conservation of Neopetrosia species in the Western Indian Ocean region.

Toxic effects of the crude methanolic extract of &lt;i&gt;Austroplenckia populnea&lt;/i&gt; leaves and fractions on the growth of &lt;i&gt;Spodoptera frugiperda&lt;/i&gt;

The aim of the current study is to assess the toxicity of &lt;i&gt;Austroplenckia populnea&lt;/i&gt; leaf extracts towards the &lt;i&gt;Spodoptera frugiperda&lt;/i&gt; population in laboratory conditions. Initially, a crude methanolic extract was obtained from dried leaves of &lt;i&gt;A. populnea&lt;/i&gt;. This extract was fractionated using hexane, ethyl acetate, and then methanol. Neonate &lt;i&gt;S. frugiperda&lt;/i&gt; larvae were isolated in plastic containers and fed (once) on corn leaf fragments treated with crude and fractions of the investigated plant extracts. The life cycle of the larvae treated with the extracts was monitored. In addition to the intake of leaves treated with the plant extracts, such variables as larval and pupal mortality rate, larval and pupal weight, and larval, pupal, and adult stage duration were evaluated. The crude, hexane, and methanol extract fractions exerted an antixenosis effect on the feeding behaviour of neonate &lt;i&gt;S. frugiperda&lt;/i&gt; larvae. A significant antibiosis effect was observed for the hexane fraction of the &lt;i&gt;A. populnea&lt;/i&gt; extract, since all of its concentrations promoted a &lt;i&gt;S. frugiperda&lt;/i&gt; larval mortality rate differing from that observed for the control. The pupal mortality rate was low, but it did not differ significantly from the mortality rate observed for the control treatment, which was nil. The larvicidal effect of the &lt;i&gt;A. populnea&lt;/i&gt; leaf extract, mainly of its hexane fraction, has indicated its potential to be used in integrated &lt;i&gt;S. frugiperda&lt;/i&gt; management programs.

Identification of Phthalates, Methyl Esters and Siloxanes through GCMS analysis, biological investigation, DFT and Molecular Docking studies on Methanolic Subfractions of Hibiscus Rosa Sinensis flower extract

Purpose: Plants are vital source of bioactive molecules, so they play a significant role in prevention and curing of various health disorders. The Hibiscus rosa sinensis, an ornate with attractive flowers, has conventionally been utilized globally as a curative agent for a wide range of ailments. Methods: In current study, we prepared crude methanolic extract of flowers of Hibiscus rosa sinensis and subjected that to in vitro biological evaluation to assess its antidiabetic and antioxidant potential. Outcomes of in vitro evaluation confirmed the excellent antidiabetic and antioxidant potency of Hibiscus flowers. Further crude methanolic extract was fractionated to hexane, ethyl acetate and methanol fractions. Among the three fractions, we subjected methanolic fraction to GCMS analysis to explore the bioactive molecules contained by this fraction. Results: The spectral data obtained from GCMS analysis revealed the existence of a total of 12 compounds including phthalates, methyl esters and siloxanes which are known to have a range of reported pharmacological activities. Successively, we subjected the identified molecules to molecular docking to assess their antidiabetic and antioxidant potential and DFT study to explore the reactivity of their molecular structures. As far as we are aware, it is the first report on the in-silico antidiabetic (128.1µM) and antioxidant (410.0µM) potential assessment of phytoconstituents of methanolic extract of floral part of Hibiscus rosa sinensis. The results revealed the H. Sinensis plat as more viable and good alternative to existing natural remedies.

Houttuynia cordata Thunb. Hexane fraction induces MDA-MB-231 cell apoptosis via caspases, ER stress, cell cycle arrest and attenuated Akt/ERK signaling

Houttuynia cordata Thunb. (HCT) is a perennial plant used in traditional Thai medicine for many centuries. This study aimed to investigate the antiproliferative effect of the hexane fraction, which has not been explored before. HCT ethanol extract (crude extract) was sequentially fractionated to obtain a hexane (H) fraction. GC-MS was used to determine the phytochemicals. The H fraction consisted of lipids, mainly α-linolenic acid and some terpenoids. MTT assay was used to determine the cytotoxic effects of H fraction in MCF-7, MDA-MB-231, NIH3T3 and PBMCs. The mode of cell death and cell cycle analysis were determined by flow cytometry. The mechanisms of cell death were defined by mitochondrial transmembrane potential (MTP) reduction and activation of caspase-3, -8 and -9. The expression levels of the Bcl-2 family, cell cycle-related, endoplasmic reticulum (ER) stress-associated proteins; and Akt/ERK signaling molecules were investigated by immunoblotting. The H fraction was toxic to MDA-MB-231 more than MCF-7 cells but not to NIH3T3 and PBMCs. The growth of MDA-MB-231 cells was inhibited through apoptosis. MTP was disrupted whereas caspase-3, -8 and -9 were activated. The expression of pro-apoptotic Bax and Bak was upregulated, while Bid and anti-apoptotic Bcl-xL proteins were downregulated. Cyclin D1 and CDK4 levels were downregulated. The cell cycle was arrested at G1. Moreover, GRP78 and CHOP elevation indicated ER stress-mediated pathway. The expression ratio of pAkt/Akt and pERK/ERK were reduced. Taken together, the molecular mechanisms of MDA-MB-231 cell apoptosis were via intrinsic/extrinsic pathways, cell cycle arrest, ER stress and abrogation of Akt/ERK survival pathways. According to the most current research, the H fraction may be used as an adjuvant in the BC treatment; however, before the anticancer strategy can be applied to patients, it is important to determine each active compound's effects in cell lines and in vivo when compared with a combined mixture.

Obtention of fatty acids and phenolic compounds from Colombian cashew (Anacardium occidentale) nut shells using pyrolysis: towards a sustainable biodiesel production

Sustainable management of non-edible agricultural residues of cashew nut production is a concern in Colombia. Therefore, this study aimed to study the fatty acid content of a pyrolytic liquid obtained from cashew nut shells (CNSs) from the Vichada region in Colombia. Transesterification of pyrolytic liquid was conducted to obtain biodiesel at the micro-scale as the first approach for this valorization route. Proximal analysis of samples was carried out using advanced analytical techniques (UHPLC-MS and CG-MS) whereas phenolic content and antioxidant activity were determined. The production yield of pyrolytic liquid was 69.15 ± 5.07% weight (wt.), at 550 °C and 2h of pyrolysis and the liquid was rich in fatty acids (∼70% wt.) and long-chain phenols (∼18% wt.). Among the phenolic compounds in liquid, mainly unsaturated C15:4 cardanol was identified (82.1 ± 5.5 mg/g), whereas the antioxidant activity of pyrolytic liquid was 0.714 ± 0.030 TE/g. Moreover, the biodiesel yield was 81% using catalyst sodium methoxide (12% v), and 50 °C and 26 min for the reaction. The obtained biodiesel in the hexane fraction was rich in methyl trans-8-octadecanoate (20.9 % wt.) and methyl palmitate (14.3 % wt.), being the representative compounds in the biodiesel. Therefore, the results indicated that thermal conversion of CNSs for obtaining biodiesel on a one-step process is a suitable strategy for the management of toxic and non-edible cashew residues. Finally, this is the first work of its kind that propose in detail the composition of pyrolytic liquid obtained from Colombian cashew nut residues under the proximate analysis approach and using advanced analytical techniques.

Investigation of Cox-2 inhibition of Laportea decumana (Roxb). Wedd extract to support its analgesic potential

Ethnopharmacological relevanceItchy leaves Laportea decumana (Roxb). Wedd is an indigenous plant in Maluku, Indonesia, and is used traditionally to treat complaints such as fatigue and joint and muscle pains. Aim of the studyTo provide scientific proof of the analgesic effect of L. decumana ethanolic extract tested in in vivo assays while investigating its bioactive phytochemicals using liquid chromatography tandem high-resolution mass spectrometry (LC-HRMS) profiling and Cox-2 inhibition assay. Materials and methodsTo investigate the analgesic activity of the ethanolic extract, assays were conducted on male mice Balb/c strain by chemical induction using acetic acid (i.p.) and heat induction (hotplate). Mice were divided into six groups consisting of six mice, i.e., the baseline group; positive control group (paracetamol 80 mg/kg BW); groups treated with extracts in dosages of 100, 200, and 400 mg/kg bodyweight (BW); and negative control group (acetic acid 0.6%, i. p.). The crude extract was partitioned with liquid–liquid fractionation to yield hexane, ethyl acetate, and water fractions. The extract and fraction were assayed for Cox-2 enzyme inhibition, and the chemical profiles were analyzed using untargeted LC-HRMS. ResultsThe analgesic assays revealed the dose-dependent effect of the extracts, of the effect of treatment with 400 mg/kg BW was not significantly different with that of paracetamol (p < 0.05). The ethyl acetate fraction showed IC50 of 19.25 μg/mL on Cox-2 inhibition (IC50 celexocib 18.48 μg/mL). LC-HRMS showed a distinctive profile of the ethyl acetate fraction compared with those in the extract and other fractions. ConclusionsThis study presents scientific evidence of the analgesic activity of the L. decumana ethanol extract given orally to experimental animals. Inflammatory inhibition plays a role in the overall analgesic mechanism by Cox-2 inhibition of the extract and all fractions. This finding is also supported by the phytochemical profiles of the extract and fractions, showing the presence of compounds reported elsewhere as anti-inflammatory activity.

Evaluation of Rhodiola rosea Root Fraction as a Functional Cosmetic Ingredient

To develop functional cosmetic material using &lt;i&gt;Rhodiola rosea&lt;/i&gt;, the biological activities such as anti-oxidant, anti-inflammatory, astringent, anti-tyrosinase, and anti-elastase activity were investigated with methanol, &lt;i&gt;n&lt;/i&gt;-hexane, chloroform, ethyl acetate, n-butanol, and water fractions of &lt;i&gt;R. rosea&lt;/i&gt; Root. DPPH and ABTS radical scavenging activities of ethyl acetate fraction were increased in a dosedependent manner; the activities were higher than other fractions. The total phenolic content of ethyl acetate fraction also showed a relatively high value of 500.485±4.848 mgGAE/g. The ethyl acetate fraction also significantly decreased LPS-induced NO production and IL-6 secretion compared with untreated control cells at above 5-100 μg/mL concentrations. The astringent activity of ethyl acetate fraction was 94.31% at a concentration of 500 μg/mL, and the activity of ethyl acetate fraction was similar to that of tannic acid. The mushroom tyrosinase inhibition abilities of hexane, ethyl acetate, and butanol were 10.44%, 17.53%, and 14.55%, respectively, at 100 μg/mL. Furthermore, an elastase inhibition rate showed a relatively high effect at 100 μg/mL, 45.46% of inhibition rate in the ethyl acetate and 45.07% of inhibition rate in the butanol fraction. These results indicate that R. rosea Root would be used as a natural material in the functional cosmetic and beauty-food industries.

Evaluation of phytochemical constituents, total phenolic contents and in vitro antioxidant activities of Mucuna pruriens fractions leaves

Mucuna pruriens is a tropical annual climbing legume plant used in traditional medicine systems to treat diabetes, arthritis, dysentery, hematinics, inflammation, and cardiovascular conditions. The present study investigates the phytochemical constituents, phenolic contents and in vitro, antioxidant potentials of M. pruriens leaf fractions using spectrophotometric methods. Phytochemical screening of hexane, chloroform, ethyl acetate, and methanol fractions of M. pruriens was determined using standard methods. The antioxidant activities of the fractions were assessed against total phenolic contents, total antioxidant contents, ferric ion reducing antioxidant power assay, and 1,1-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity using standard protocols. Phytochemical screening showed that only the ethyl acetate fraction of M. pruriens leaves contain all the phytochemical constituents tested for, saponins, tannins, flavonoids, cardiac glycosides, anthraquinones, terpenoids, steroids, alkaloids, phenol. The activities of the fractions against total phenolic contents were high in ethyl acetate fraction (1.958±0.24 mg GAE/g), while chloroform fraction gave the lowest total phenolic contents (1.476±0.19 mg GAE/g). On the contrary, chloroform fraction showed more significant total antioxidant contents (2.078±0.80 mg ASCE/g). The methanol fraction demonstrated higher ferric ion-reducing antioxidant power (4.866±1.19 mg ASCE/g). The methanol fraction scavenging activity against DPPH radical exhibited a lower IC50 value of 404.92 (μg/mL) relative to other fractions. It is concluded that ethyl acetate and methanolic fractions of Mucuna pruriens leaves, which contain large amounts of phenolic compounds, exhibited high antioxidant activities. These in vitro assays indicated that Mucuna pruriens leaves fraction is a significant source of natural antioxidants, which might help prevent the progress of various oxidative stresses.&#x0D;

HPLC-PDA Method for Quantification of Bioactive Compounds in Crude Extract and Fractions of Aucklandia costus Falc. and Cytotoxicity Studies against Cancer Cells.

Aucklandia costus Falc. (Synonym: Saussurea costus (Falc.) Lipsch.) is a perennial herb of the family Asteraceae. The dried rhizome is an essential herb in the traditional systems of medicine in India, China and Tibet. The important pharmacological activities reported for Aucklandia costus are anticancer, hepatoprotective, antiulcer, antimicrobial, antiparasitic, antioxidant, anti-inflammatory and anti-fatigue activities. The objective of this study was the isolation and quantification of four marker compounds in the crude extract and different fractions of A. costus and the evaluation of the anticancer activity of the crude extract and its different fractions. The four marker compounds isolated from A. costus include dehydrocostus lactone, costunolide, syringin and 5-hydroxymethyl-2-furaldehyde. These four compounds were used as standard compounds for quantification. The chromatographic data showed good resolution and excellent linearity (r2 ˃ 0.993). The validation parameters, such as inter- and intraday precision (RSD < 1.96%) and analyte recovery (97.52-110.20%; RSD < 2.00%),revealed the high sensitivity and reliability of the developed HPLC method. The compounds dehydrocostus lactone and costunolide were concentrated in the hexane fraction (222.08 and 65.07 µg/mg, respectively) and chloroform fraction (99.02 and 30.21 µg/mg, respectively), while the n-butanol fraction is a rich source of syringin (37.91 µg/mg) and 5-hydroxymethyl-2-furaldehyde (7.94 µg/mg). Further, the SRB assay was performed for the evaluation of anticancer activity using lung, colon, breast and prostate cancer cell lines. The hexane and chloroform fractions show excellent IC50 values of 3.37 ± 0.14 and 7.527 ± 0.18 µg/mL, respectively, against the prostate cancer cell line (PC-3).