Bioactive Methanol Extract Research Articles

Phytochemical analysis and in vitro biological activity assessment of extracts from Micromeria myrtifolia

ABSTRACT In this study, the antioxidant capacity, the effect on anti-acetylcholinesterase and anti-urease enzyme inhibition, and kidney stone-reducing effects of different extracts from M.myrtifolia. In addition, the phytochemical content of the bioactive methanol extract was analyzed by HPLC-DAD. The methanol extract showed strong DPPH (IC50:0.036 mg/mL) radical scavenging, Cu (II) ion (6.535 mmTroloxE) and Fe3+ (6.476 FeSO4) ion reducing antioxidant activity compared to other extracts. The methanol extract contained the highest amount of phenolic (400 mgGAE) and flavonoid (2.519 QuE) compounds. Moreover, the methanol extract had significant anticholinesterase (78.935%), anti-urease (71.014%) and calcium oxalate anti-crystallization (18.22%, 37.427%) activities. The findings show that all extracts exhibited varying amounts of antibacterial activity against Staphylococcus aureus. The bioactive methanol extract contained rosmarinic acid (31.35 µg), quercetin (12.99 µg) and chlorogenic acid (3.55 µg). The analyzed compounds are thought to contribute significantly to the pharmacological effect of the methanol extract.

In vitro antioxidant and anticholinesterase activity of Aegle marmelos fruit: implications for Alzheimer’s disease treatment

AbstractBackgroundIncreased antioxidant and anticholinesterase activity are key pathological hallmarks of Alzheimer’s disease (AD). In various Ayurvedic formulations Aegle marmelos (AM) belonging to family Rutaceae have traditionally been used as a brain tonic. However, there was no scientific validate proof of it. Hence, the present study was aimed to identify bioactive extract/fraction/constituent of AM fruit having potential to treat AD.MethodFour different crude extracts of AM fruits by using different solvents on basis of increasing polarity‐such as petroleum ether, chloroform, methanol and distilled water‐by successive Soxhlet extraction were prepared and evaluated for in‐vitro antioxidant and anticholinesterase inhibition assay by using DPPH and Ellman’s assay respectively. Most bioactive extract was further subjected to fractionation by using solvent‐solvent partioning followed by flash chromatography for further sub fractionation of most bioactive fraction. Total phenolic (TPC) and flavonoid content (TFC) was also calculatedResultComparative evaluation of all four prepared extracts for in‐vitro antioxidant and anticholinesterase inhibition assays showed that the methanol extract (ME) was most bioactive. Furthermore, bioactivity‐guided fractionation of this bioactive ME revealed that Dichloromethane (DCM) fraction evinced highest antioxidant and anticholinesterase activity with IC50 value 74.56±1.43 µg/ml and 47.65±2.76 µg/ml due to presence of higher phenolic and flavonoid content. However, further sub‐fractionation of DCM fraction showed significantly higher IC50 values thus decreased activity.ConclusionThis preliminary study clearly suggested that the DCM fraction from the ME of A. marmelos may prove to be potential candidate for the treatment of AD may be due to synergistic effect of phenolic and flavonoid content present in it. However, to validate the neuroprotective effect of DCM fraction of AM fruit against AD further exploration at cellular and molecular level is required to underlying its pharmacological mechanism as well as identification of the active phytoconstituents responsible for anti‐Alzheimer’s effect.

Greener approach for the isolation of oleanolic acid from Nepeta leucophylla Benth. Its derivatization and their molecular docking as antibacterial and antiviral agents

In the present study bioactive methanolic extract along with chloroform and hexane extracts obtained from shade dried leaves of the Himalayan aromatic medicinal plant Nepeta leucophylla Benth. Were screened for the presence of triterpenoids, especially oleanolic acid (OA). Total three compounds oleanolic acid, squalene and linoleic methyl ester were isolated from methanol extract. The percentage yield of OA was 0.11%. Out of these three, OA is more bioactive and was further subjected to derivatization using greener Ultrasonication method. Total three derivatives (3-Acetyl oleanolic acid, 3-Phthaloyl oleanolic acid and 3-Oxo oleanolic acid) were synthesized with 91.16%, 93.98%, and 83.6% respectively. Further, the antioxidant potential of OA and its derivatives were evaluated using DPPH assay which suggested that the 3-Phthaloyl oleanolic acid exhibits highest antioxidant potential with 40.83 ± 1.14% inhibition. OA and its derivatives were screened in-silico antibacterial potential against three bacterial pathogens (E-coli, M. tuberculosis and S. aureus) and antiviral potential against Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2), Human immunodeficiency virus (HIV) and H1N1 influenza virus. The in-silico results suggested that 3-phthaloyl oleanolic acid showed best H-bonding with FtsA (Staphylococcus aureus), enoyl acyl reductase (E. coli) and arabinosyl transferase (Mycobactrium tuberculosis). 3-Phthaloyl oleanolic acid also showed best H-Bond interactions with the target proteins hemagglutinin (H1N1) and reverse transcriptase (HIV), whereas, oleanolic acid exhibited the best interactions with RNA dependent RNA polymerase (SARS-CoV-2) and thus could be considered for further in vitro studies.

Molecular Docking andin silicoEvaluation of Phytochemicals of Bioactive Methanolic Extract ofIpomoea mauritianaJacq. as Anti-Bacterial Agents

Antibacterial treatment has grown difficult due to the increasing growth in bacterial infections, as well as their tolerance to most first-line antibiotics. This is a severe danger to the world’s human health in the 21st century, necessitating further research to identify drugs with improved antibacterial effects and broad-spectrum functions. This study aimed to discover anti-bacterial agents through the molecular docking and in silico approach. Most responsive thirty (32) compounds on UPLC-Q-TOF/MS analysis were selected from our previous report to get the hit compound(s) against inhibition of cell wall synthesis, inhibition of protein synthesis, interference with nucleic acid synthesis, inhibition of a metabolic pathway, inhibition of membrane function and inhibition of adenosine triphosphate (ATP) synthase. From the molecular docking results, we afforded six compounds for cell wall synthesis protein, four compounds for protein synthesis protein, five for nucleic acid synthesis protein, three for metabolic pathway protein, four for membrane function protein and three for ATP synthase protein which eventually undergoes the pharmacokinetic and drug-likeness properties to obtain lead compound(s). Finally, we discovered that compounds Turpinionosides B, Polydatin, Ledebouriellol, and Pterodontoside A have the strongest binding interactions with cell wall synthesis, inhibition of protein synthesis and inhibition of metabolic pathway synthesis, interference with nucleic acid synthesis and inhibition of ATP synthase, inhibition of membrane function proteins, respectively. These compounds have the potential to become an anti-bacterial therapeutic candidate due to their promising pharmacological properties.

Antioxidant, anti-tyrosinase and anti-inflammatory activities of 3,5-dihydroxy-4′,7-dimethoxyflavone isolated from the leaves of Alpinia nigra

BackgroundOverexposure to sunlight and high UV radiation leads to uncontrolled melanin production in the skin, leading to hyperpigmentation and inflammatory diseases. Therefore, there is a need to discover safe and highly prospective drugs from natural sources. Previous studies have shown that leaves and rhizome extract of Alpinia nigra possesses antioxidant and anti-inflammatory activities. Among the components of the most bioactive methanol and ethyl acetate extract, flavones seem to be the most promising compound. AimTo evaluate the antioxidant, anti-tyrosinase and anti-inflammatory efficacy of 3,5-dihydroxy-4′,7-dimethoxyflavone (DHDM). MethodsThe flavone DHDM was isolated from A. nigra ethyl acetate leaf extract and characterized by multi spectroscopic techniques. The toxicity of DHDM was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The antioxidant and reactive oxygen species (ROS) inhibitory activity were determined by 2,2-Diphenyl-1-picrylhydrazy (DPPH) assay and by flow cytometry. Further, the anti-tyrosinase activity was evaluated by in silico (molecular docking), fluorescence, and enzyme kinetics studies. The anti-inflammatory activity was demonstrated by western blotting. ResultsThe cell viability assay with DHDM showed no inhibition at ≤200 µM concentration in THP-1 (human macrophage) and ≤80 µM in HaCaT (human keratinocyte) cell lines. Additionally, strong antioxidant properties and reduced ROS generation suggested prevention from oxidative damage and skin aging. Strong binding of DHDM to tyrosinase leading to the conformational change was revealed by fluorescence quenching studies and molecular docking. Further, the anti-tyrosinase activity of the isolated compound was observed at 42.5 µM (IC50). Moreover, DHDM inhibited the expression of anti-inflammatory proteins like TNF-α and NF-κB. ConclusionThe DHDM demonstrates the efficient antioxidant, anti-tyrosinase and anti-inflammatory potential, which could be developed for obtaining a new drug for the management of hyperpigmentation and inflammatory-related diseases.

Isolation, characterization, and chromatographic estimation of esculin: A potential fluorescent marker from Launaea pinnatifida cass

Background: Launaea pinnatifida (Willd.) Sch. Bip ex Kunze (Asteraceae) is an Indian Ocean intertidal sand dune herb that is useful as traditional medicine. The plant is one of the species of traditional Indian Ayurvedic herb “Gojihva.” Objectives: The present study was performed to isolate, characterize, and estimate a potential marker compound from L. pinnatifida. Materials and Methods: The leaves of the plant were subjected to successive solvent extraction with the help of solvents of varying polarity. Based on bioactivity studies performed using these extracts, the most bioactive methanolic extract was further subjected to vacuum liquid chromatography using chloroform and methanol in varying proportions. A phytoconstituent was isolated from fraction chloroform: methanol (90:10), elucidated to be Esculin based on its analytical (FTIR, liquid chromatography–mass spectrometry, 1H nuclear magnetic resonance, and ultraviolet) data. For the quantitative analysis of esculin in L. pinnatifida leaves, simple and validated high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) methods were developed. Results: The purity of isolated as well as purchased standard esculin was found to be 97.66% and 98.2%, respectively, by HPLC, whereas the yield of esculin in the methanolic extract of leaves was found to be 1.40% w/w by HPTLC. Conclusion: Esculin has been reported for the first time in this plant and can be considered a potent phytochemical marker. It can also be useful for the standardization and quality control of L. pinnatifida formulations.

Evaluation of Acacia auriculiformis Benth. leaves for wound healing activity in type 2 diabetic rats

Background: Acacia auriculiformis Benth. is a perennial shrub which has been traditionally known to treat various medical complications such as sore eyes, allergy, rheumatism, sore eyes, and rashes. Objectives: The current study is designed to decipher a shred of clinical confirmation and biochemical support for wound-healing efficacy of methanolic leaves extract of A. auriculiformis. Materials and Methods: We prepared the leaves extract in different solvents, i.e., petroleum ether, chloroform, ethyl acetate, acetone, butanol, and methanol. The in vitro studies were carried out on the above leaf's extracts using α-glucosidase and α-amylase inhibition in vitro antidiabetic assays. Animal model of streptozotocin-induced diabetes was used in the 0-, 7-, and 15-day studies, respectively. The comparison study was carried out in diabetic wound control in respect of the period of epithelialization, %wound contraction, and hydroxyproline content in the excision wound model. However, the breaking strength parameter was used to calculate healing potential in the incision wound model. The bioactive methanol extract was subjected to LC-MS/MS analysis to characterize the phytoconstituents responsible for pharmacological activity. Results: The methanolic leaves extract showed the highest percentage inhibition of 94.259% and 95.259% in α-glucosidase and α-amylase in vitro antidiabetic assays, respectively. The high content of collagen fibers and stronger epithelial cells growth were observed in histopathological studies of hydrogel containing methanolic leaves extract as compared to the diabetic wound control and standard. Subsequently, for investigating the biological impact upon live cells, cytotoxicity study was tested in different cell lines (A549, HEK-293, and MCF-7). Cytotoxic results showed that greater than 75% of cells were visible in all the cell lines, which gives the confirmation of biocapability of the extract. The LC-MS/MS results revealed the presence of compounds such as β-sitosterol, lupeol, stigmasterol, and quercetin. Conclusion: A. auriculiformis is a potent medicinal plant that can be further utilized as a complementary and alternative therapy for the treatment of diabetes-induced wounds and the management of oxidative stress and diabetes.

Potent bioactive methanolic extract of wild orange (Citrus macroptera Mont.) shows antioxidative, anti-inflammatory, and antimicrobial properties in in vitro, in vivo, and in silico studies

BackgroundThere is always an increasing demand for natural remedies from natural sources which can substitute the synthetic therapeutic drugs and lessen their side effects. The present study aims to investigate the antioxidant, anti-inflammatory, antimicrobial properties and in silico docking study of Citrus macroptera leaf (CML) extract in both in vivo and in vitro aspect.Material and methodsThe antioxidant and anti-inflammatory potential of crude extract was investigated in vitro and in vivo on Wistar albino rat. The antioxidant potentiality also investigated on HepG2 cell line. Antimicrobial activity was evaluated against Staphylococcus sp. and Klebsiella sp. Chemical compounds of the crude extract were identified by GC-MS analysis. In silico docking was also done against NF-ҡB protein.ResultsAt 200 μg/ml concentration, CML significantly scavenges reactive oxygen species (ROS) which was generated on HepG2 cell line. CML showed 71% anti-inflammatory activity (p ≤ 0.001) against carrageenan-induced paw edema in albino Wistar rats. CML extract is very effective against staphylococcus sp. than Klebsiella sp. In the docking analysis, the proximadiol and menthone had − 5.6 kcal/mol and − 5.7 kcal/mol binding affinity with the protein NF-ҡB.ConclusionIn the present work, CML provided notable antioxidant, anti-inflammatory, and antimicrobial activity. This activity was confirmed by both in vitro and in vivo followed by in silico docking technique. Overall, the experimental results presented in this study suggest that crude extract of CML could be used as a promising antioxidant and anti-inflammatory candidate with potential benefits.

Insect Antifeedant Components of Senecio fistulosus var. fistulosus-Hualtata.

From a bioactive methanolic extract of Senecio fistulosus, the antifeedant effects of the alkaloidal and non-alkaloidal fractions were tested against the insects Spodoptera littoralis, Myzus persicae and Rhopalosiphum padi, with the non-alkaloidal fraction being antifeedant. The phytochemical study of the non-alkaloidal fraction of S. fistulosus, resulted in the isolation of four compounds, two 9-oxo-furanoeremophilanes (1, 2), an eremophilanolide, 1β,10β-epoxy-6-acetoxy-8α-hydroxy-eremofil-7(11)-en-8β,12-olide (3) and a maaliol derivative (4). The alkaloidal fraction yielded two known pyrrolizidine alkaloids (5, 6). Compounds 1, 3 and 4 are new natural products. Furanoeremophilane 2 was a strong antifeedant against S. littoralis and maaliane 4 inhibited the settling of M. persicae.

Neuropharmacological Profile of Fractions of Actaea acuminata H. Hara Roots

The methanol extract of Actaea acuminata roots have been previously reported to exhibit significant antianxiety, anticonvulsant and antidepressant activities. In present study fractionation of crude bioactive methanol extract was carried out using different solvents employing standard procedure. Various fractions were evaluated for antianxiety, anticonvulsant and antidepressant activities using elevated plus maze model (EPM), maximal electroshock-induced convulsions model (MES) and forced / despair swim test (FST) respectively. Successive partitioning of bioactive methanol extract was done with ethyl acetate and 1-butanol. The ethyl acetate fraction (EAF; 100 mg/kg), 1-butanol fraction (BF; 25 mg/kg) and remaining methanol extract (RME; 70 mg/kg) were subjected to neuropharmacological activities. The EAF significantly enhanced entries and average time spent in open arms; significantly decreased duration of MES-induced tonic extension phase and significantly decreased duration of immobility time of rats in comparison to control. BF and RME did not exhibit any neuropharmacological activity. Qualitative chemical tests confirmed presence of alkaloids and polyphenols in EAF. It is finally concluded that alkaloids and/or polyphenols are bioactive constituents of A. acuminata which are responsible for neuropharmacological activities.

Bacillus amyloliquefaciens induces production of a novel blennolide K in coculture of Setophoma terrestris

The discovery of known bioactive chemical leads from microbial monocultures hinders the efficiency of drug discovery programmes. Therefore, in recent years, the use of fungal-bacterial coculture experiments has gained considerable attention due to their ability to generate new bioactive leads. In this work, fungal strain Setophoma terrestris was cocultured with Bacillus amyloliquifaciens to discover novel bioactive compounds. The bioactive methanolic coculture extract was chosen for the isolation of compounds by chromatographic methods. The isolated compounds were characterized by NMR and mass spectrometric techniques. Coculture extract has resulted in the production of five blennolides. The novel compound, blennolide K was found active against PC-3 (prostate) and MCF-7 (breast) cell lines with an IC50 value of 3·7±0·6 and 4·8±0·4μmoll-1 respectively. Furthermore, the nuclear morphology study in PC-3 cells after treatment with blennolide K, demonstrated chromatin condensation, formation of apoptotic bodies and shrinkage of cells. To our knowledge, only few studies have reported the induction of bioactive compounds by coculture having long-distance inhibition morphology. This is principally due to the low occurrences of such morphology. Our study demonstrates the impact of coculture on production of new chemical leads in drug discovery programmes.

Activity guided isolation of chemical constituents from the biologically active methanol extract of Euphorbia schimperi C. Presl

In this study we investigated the chemical constituents of bioactive methanol extract of Euphorbia schimperi C. Presl. For this the methanol extract was fractionated into 20, 40, 60, 80% MeOH in CHCl3, and 100% MeOH fractions respectively by vacuum liquid chromatography. Excision wound surface of the animals were topically treated with these fractions at a dose of 100 mg/kg body weight for twenty days. Povidone-iodine ointment was used as a reference drug. Wound contraction measurement and period of epithelialization were used to assess the effect of fractions on wound repairing. The 100% MeOH fraction treated animals achieved significant (p < 0.001) value by showing 100% wound contraction and minimum period of epithelization (17.75±0.47) on the 20th day as compared to standard drug treated animals on the same day. The active 100% MeOH fraction was subjected to various chromatographic techniques led to the isolation of miquelianin (1), kaempferol 3-O-glucuronide (2) and quercitrin (3). Compounds (1-3) were isolated from this plant for the first time. KEY WORDS: Euphorpia schimperi C. Presl, Wound healing, Chromatographic techniques, Chemical constituents Bull. Chem. Soc. Ethiop. 2017, 31(3), 471-479. DOI: http://dx.doi.org/10.4314/bcse.v31i3.11

Cytotoxic lanostane triterpenoids from the fruiting bodies of Piptoporus betulinus

Chemical investigation of a bioactive methanolic extract of the fruiting bodies of Piptoporus betulinus led to the isolation of five previously undescribed lanostane triterpenoids named piptolinic acids A−E, as well as five known lanostane triterpenoids. Their structures were elucidated on the basis of 1D and 2D NMR spectroscopic and HRESIMS analysis. Piptolinic acid A with an unusual moiety (3-hydroxy-4-methoxycarbonyl-3-methylbutyryloxy) at C-3 exhibited comparable cytotoxic activity against human promyelocytic leukemia cell line HL-60 (IC50 = 1.77 μM) and human acute monocytic leukemia cell line THP-1 (IC50 = 8.21 μM) to those of positive control, fluorouracil (IC50 = 6.38 and 4.41 μM, respectively).

Ameliorative potential of Alstonia scholaris (Linn.) R. Br. against chronic constriction injury-induced neuropathic pain in rats

BackgroundAlstonia scholaris commonly known as ‘Saptaparni’ is an Indian traditional medicinal plant used in Ayurveda. It is commonly used to treat various disorders like asthma, bronchitis, diarrhea, dysentery and malaria. In folklore medicine the milky juice of the plant is applied on wounds and ulcers to treat pain, ear ache and also in rheumatic pains.AimThe present study was designed to investigate the potential of A. scholaris R. Br. in chronic constriction injury of sciatic nerve (CCI) induced neuropathic pain in rats.MethodsPeripheral neuropathy was induced by chronic constriction injury of sciatic nerve. The behavioral parameters like mechanical and thermal hyperalgesia and cold allodynia were assessed on the 14th day. Tissue parameters like total protein, thiobarbituric acid reactive substances, reduced glutathione, myeloperoxidase, total calcium and TNF-α were assessed to check biochemical changes. Chloroform and methanol extract of A. scholaris leaves (100 and 200 mg/kg) and pregabalin (10 mg/kg, as positive control) were administered orally for 14 consecutive days starting from the day of surgery.ResultsCCI resulted in significant development of mechanical hyperalgesia, heat hyperalgesia and cold allodynia along with alteration in the biochemical changes. Administration of methanol extract at 200 mg/kg significantly attenuated the CCI induced change in nociceptive threshold and biochemical changes which was comparable to that of pregabalin. The high-performance liquid chromatography (HPLC) of the bioactive methanol extract revealed the presence of different types of flavonoids such as gallic acid, catechin, epicatechin, ellagic acid and kaempferol, in which kaempferol was observed to be in higher concentration.ConclusionMethanol extract (200 mg/kg) of A. scholaris showed the ameliorative effect in CCI induced neuropathic pain which may be due to the presence of kaempferol and attributed to its anti-oxidative and anti-inflammatory properties.

Screening of Antidepressant Activity and Marker-based Standardization of Baptisia tinctoria (L.) R. Vent.

Baptisia tinctoria (Wild indigo; Fabaceae) has long tradition of use in Indian systems of medicine for the treatment of depression. This investigation deals with the evaluation of the antidepressant activity of various extracts and fractions of B. tinctoria roots and estimation of the content of bioactive principle using thin-layer chromatography densitometry. Roots of B. tinctoria were successively extracted to obtain n-hexane, chloroform, methanol and water extracts. The chloroform, methanol and water extracts were screened for antidepressant activity at doses of 200 or 400 mg/kg, p.o., in mice subjected to forced swim test. Methanol extract showed significant activity at a dose of 400 mg/kg. The methanol extract was further fractionated successively to obtain ethyl acetate and 1-butanol fractions. Antidepressant activity of these fractions was assessed using forced swim test at the doses of 90 and 80 mg/kg, p.o., respectively. The ethyl acetate fraction showed significant antidepressant activity at the dose of 90 mg/kg in mice. Further, specific antidepressant activity without psychostimulant effects of bioactive methanol extract and ethyl acetate fraction was confirmed through evaluation of locomotor behaviour in mice using open field test. Phytochemical screening showed the presence of flavonoids as major class of phytoconstituents in the methanol extract. Comparative thin-layer chromatography fingerprint studies confirmed the presence of hesperitin in methanol extract. Hesperitin was used as a chemical marker to standardize B. tinctoria roots using validated thin-layer chromatography densitometric method and the content of hesperitin was found to be 0.0085% w/w.

Evaluation of Antianxiety Potential of Four Ganoderma (Agaricomycetes) Species from India in Mice.

The genus Ganoderma consists of widespread polypore mushrooms that have traditionally been used to reduce stress and anxiety. However, scientific evidence for this is not adequate. Hence, this study was designed to investigate the anxiolytic potential of G. applanatum, G. brownii, G. lucidum, and G. philippii collected from Uttarakhand, India. Various extracts of dried, powdered basidiocarps were prepared using different solvents-namely, petroleum ether, chloroform, methanol, and distilled water-by successive Soxhlet extraction. All the extracts were tested for antianxiety activity using the elevated plus maze (EPM) model in Swiss albino mice. The results showed that the methanol extract of G. lucidum at a dose of 200 mg/kg, administered orally, shows a significant increase in the average time spent in the open arms of the EPM when compared with the control; this was comparable to the effect of the standard drug (diazepam, 2 mg/kg by mouth). This bioactive methanol extract was subjected to bioactivity-guided fractionation. The results show that the n-butanol fraction of the methanol extract evinced significant antianxiety activity at a dose of 100 mg/kg. This fraction showed the presence of phenols and flavonoids and thus was standardized with respect to total phenol content and total flavonoid content. The antianxiety activity may be the result of the phenols/flavonoids present. This study clearly demonstrated that the n-butanol fraction from the methanol extract of G. lucidum can be developed as source of new anxiolytic agents.

Screening of Antianxiety Activity of Abies pindrow Royle Aerial Parts

Abies pindrow Royle (Himalayan Silver Fir; Pinaceae) has been traditionally used for the treatment for anxiety but the plant has not been systematically investigated to validate its traditional claims. Thus, it was planned to investigate antianxiety activity of various extracts and fractions of A. pindrow aerial parts using elevated plus maze model (EPM). Properly identified A. pindrow aerial parts were successively and exhaustively extracted using solvents in increasing order of polarity viz., n-hexane, chloroform, methanol and water. All crude extracts were subjected to antianxiety activity at the doses of 100, 200 or 400 mg/kg, p.o. in mice. Efficacy of A. pindrow was statistically compared with the standard anxiolytic drug, diazepam (2 mg/kg, i.p.). Amongst various extracts, chloroform and methanol extract exhibited significant antianxiety activity with respect to control, and statistically equivalent to the standard drug at the dose of 200 and 400 mg/kg, respectively. The bioactive methanol extract was partitioned successively using solvents in increasing order of polarity viz., n-hexane, ethyl acetate and 1-butanol. All fractions were also subjected to antianxiety activity at the doses of 25 or 50 mg/kg, p.o. in mice. Only ethyl acetate fraction (EAF) exhibited significant antianxiety activity equivalent to the standard drug at the dose 50 mg/kg. Present studies scientifically validated traditional claims of A. pindrow for antianxiety activity. Flavonoids of A. pindrow aerial parts have been suggested to play role in treating anxiety disorders.

In vitro cytoprotective potential of ornamental Harpullia pendula highlighting its medicinal value

Harpullia pendula F., family Sapindaceae is a small to medium sized rainforest tree. The plant is known as tulipwood or tulip lancewood and is mainly cultivated as ornaments [1]. Leaves and seeds from H. pendula were successively extracted with petroleum ether, chloroform (CHCl3), and methanol (MeOH), the total aqueous methanol extract of the leaves was also prepared. The methanol extract was fractionated on Diaion HP20, compounds were isolated and purified by paper chromatography, Sephadex column and HPLC. The chloroform extract was chromatographed on silica gel column. The study aimed to investigate the direct cell-free radical scavenging activity using the micro-scaled DPPH assay [2] whereas the cell-based anti-oxidative stress activity was tested by the ability to protect murine hepa1c1c7 liver cells against the cell damage induced by t-butyl hydroperoxide (TBHP), assessed by neutral red uptake assay [3]. The total alcoholic extract from the leaves showed potent scavenging of DPPH radical with EC50 of 8.4 µg/ml. The same extract also showed the most potent intracellular protection (71% viability to control) of hepa1c1c7 cells against the strong cell death induced by TBHP. The potency of the fractions obtained by successive extraction was in the order MeOH> aqueous MeOH> CHCl3 recording 30%, 25% and 4% cell viability, respectively, where EC50 against DPPH was calculated as 3.4, 5.3 and 48.6 µg/ml respectively. Column chromatography of the bioactive methanol extract allowed the isolation and identification of flavonoid glucosides and galloyl compounds. Triterpenoids were also isolated from chloroform extract. To the best of knowledge, these data represent the first report of cytoprotective potential of H. pendula leaves extracts and warrant further bioassay-guided fractionation to reveal the bioactive phytochemical(s).

Cytotoxic Long-chain Alkene and Terpene Isolated from the Methanol Extract of the Air-dried Leaves of Pipturus arborescens C.B. Rob

A long-chain alkene (1-hexacosene) and a terpene were obtained from the bioactive methanol extract of the air-dried leaves of Pipturus arborescens (Link) C.B. Rob (“Handalamay”) collected from Poblacion, Kapatagan, Lanao del Norte. The compounds were isolated through extraction with ethanol, sequential partitioning with water-chloroform and hexane-90% methanol followed by fractionation and purification with repeated gravity column chromatography using gradient mixtures of hexane: ethyl acetate (1:1 ratio). The structures of the isolates were elucidated on the basis of spectroscopic data: 1 H-and 13 C-NMR, 1 H- 13 C Heteronuclear Multiquantum Correlation (HMQC), 1 H- 13 C Heteronuclear Multibond Correlation (HMBC), 1 H- 1 H Homonuclear Correlation Spectroscopy (COSY), Infra-Red Spectroscopy (IR), Ultraviolet-Visible Spectroscopy (UV-VIS) and Mass Spectrometry (MS). The isolated compounds were shown to possess moderate cytotoxic activity against the brine shrimp Artemia salina (Leach).

Anti-Helicobacter pylori activity of Abelmoschus esculentus L. Moench (okra): An in vitro study

The anti-Helicobacter pylori activity of the methanol and hexane extracts of Abelmoschus esculentus L. Moench (okra) dried fruits were evaluated on forty-one clinical isolates and a standard ATCC 43504 strain by the use of agar well diffusion technique. The methanol extract of A. esculentus showed A. esculentus L. Moench (okra) dried fruit had inhibitory effects against Helicobacter strains; with diameters zone of inhibition between 13 and 28 mm on 32 out of the 42 isolates tested. No noticeable zone of inhibition was observed from the hexane extract of the tested plant on all the H. pylori strains tested. The bioactive methanol extract of A. esculentus demonstrated A. esculentus L. Moench (okra) dried fruit had minimum inhibitory concentration (MIC) values of 70 to 85 mgml-1 on selected susceptible strains except H. pylori ATCC 43504 which had MIC value of 250 mgml-1. The time-kill study of the methanol extract of A. esculentus on H. pylori BAA009, H .pylori BAA026 and H. pylori ATCC 43504, revealed a decline in the surviving population of the organisms after 8 h of exposure to the methanol extracts of A. esculentus L. Moench dried fruit at doses equivalent to MIC, 2 × MIC and 4 × MIC, and a total kill of the population at 24 h. Therefore, alternative antimicrobial agents may be isolated from further bioassay-guided fractionation of edibles such as A. esculentus L. Moench for the treatment of H. pylori infections, especially as they are readily available. Key words: Anti-Helicobacter pylori, Abelmoschus esculentus, fruit, kill kinetics, methanol, hexane.