Evaluation of antioxidant activity of Mimosa pudica L. extracts

In the Philippines,Cycasspp. are found in Luzon Island particularly in Pampanga, Batangas, Bataan and Isabela provinces.. In this study, the bioactive potentials of the crude methanolic, ethanolic, ethyl acetate and chloroform leaf extracts ofCycas riuminianaPorte ex Regel were investigated. Based on the results of the four solvents used, the best extraction solvent for the phytochemicals is ethanol, followed by methanol, ethyl acetate and chloroform. The ethanolic and methanolic leaf extracts showed comparable antioxidant activity. The chloroform and ethyl acetate extracts also have comparable antioxidant activity but significantly lower than both methanolic and ethanolic extracts. However the greatest antimicrobial activity was exhibited by the ethyl acetate extract, followed by chloroform, methanolic and ethanolic extracts. The variation and similarity in the antioxidant and antimicrobial activities of the different extracts can be attributed to different mechanisms of interactions, namely, independent joint action, additive, synergistic, competitive or antagonistic interactions, among the bioactive compounds present in the crude extracts. Further studies are needed to elucidate the structure of the different phytochemicals present in the leaf extracts of Arayat Pitogo (C. riuminianaPorte ex Regel) and the specific mechanisms of interaction among these phytochemicals. SUMMARY The extracts were tested for the presence (trace, moderate or abundant amounts) of flavonoids, saponins, tannins, triterpenes, alkaloids, sterols and glycosides. The ethanolic extract was positive for all phytochemicals screened with sterols, flavonoids, glycosides and tannins being abundant, alkaloids being moderate and triterpenes and saponins in trace amounts. The methanolic extract was also positive for all constituents but in trace amounts, except for flavonoids which were abundant. The ethyl acetate extract contained abundant sterols, moderate alkaloids and trace amounts of saponins, glycosides and tannins. Finally, the chloroform extract contained abundant sterols, and trace amounts of alkaloids, saponins and glycosides. The radical scavenging assay revealed that the highest percent inhibition was obtained for the ethanolic leaf extract (60.53±0.7801%), followed by methanolic extract (59.92±3.160%), chloroform extract (50.17±4.779%) and ethyl acetate extract (47.25±3.759%). In terms of antibacterial activity, the ethyl acetate extract registered the highest inhibition against the three test organisms, namely,Staphylococcus aureus, Bacillus subtilisandEscherichia coli. The chloroform extract inhibitedS. aureusandB. subtilis. The methanol extract inhibitedS. aureusonly. Finally, the ethanolic extract failed to inhibit any of the test organisms despite its abundant phytochemicals and high antioxidant activity. In terms of antifungal activity, the different extracts inhibitedCandida albicanswith the ethyl acetate and chloroform extracts showing a high degree of inhibition followed by the methanolic and ethanolic extracts. However, none of the extracts showed any bioactivity againstAspergillus niger.

Background Ixora species are perennial shrubs and flowering plants belonging to the family Rubiaceae. The leaf and flower parts of Ixora coccinea (I. coccinea)andIxora alba (I. alba) were aimed at isolating their active fractions. The present study was to determine in vitro antitumor activity against malignant melanoma cell lines for phytosome formulation. Materials and methods Two species, I. coccinea (red flowers and leaves) and I. alba (white flowers and leaves), were selected, and this study focused on determining the active fraction by comparing the in vitro antimicrobial and antioxidant potentials of petroleum ether, chloroform, ethyl acetate, and hydroalcoholic (ethanol:water, 70:30 v/v) extracts. The identified potent extract was subjected to in vitro anticancer activity in malignant melanoma cell lines. Results A phytochemical study revealed phytosterols, flavonoids, proteins, amino acids, alkaloids, carbohydrates, phenols, tannins, and diterpenes. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was used to evaluate the antioxidant effect of I. coccinea and I. alba leaf and flower extracts. In the DPPH assay, I. coccinea flower hydroalcoholic extract (ICFHA) had an IC50 value of 248.99 µg/mL, and I. coccinea leaf hydroalcoholic extract (ICLHA) had an IC50 value of 268.87 µg/mL. These two extracts had a lower value with a higher antioxidant effect. In the total antioxidant assay, I. coccinea leaf ethyl acetate extract (ICLEA) and I. coccinea leaf chloroform extract (ICLCE) have 77.4 ± 0.05 and 68.9 ± 0.03 mg of ascorbic acid equivalent per gm of extract, respectively. These two extracts exhibited a high antioxidant effect. The antimicrobial potential was evaluated using selected bacterial and fungal strains using the agar-well diffusion method. Petroleum ether and chloroform extracts of I. coccinea and I. alba leaves and flowers did not possess antimicrobial activity with any of the bacterial or fungal strains. An ethyl acetate extract and a hydroalcoholic extract of I. coccinea leaves and flowers showed antimicrobial activity against Enterococcus faecalis, Candida albicans, and Staphylococcus aureus. An ethyl acetate extract of I. coccinea flower and a hydroalcoholic extract of I. alba leaf showed a significant zone of inhibition when compared with standard chloramphenicol for all three selected strains, which may be due to the presence of active phytoconstituents. ICLHA showed a MIC of ≤300 µg/mL for Enterococcus faecalisandStaphylococcus aureusand ≤400 µg/mL for Candida albicans microbial strains. The high total flavonoid content was reported in ICLEA at 771.31 µg/mL and in I. coccinea flower ethyl acetate extract (ICFEA) at 694.69 µg/mL. High-performance thin layer chromatography (HPTLC) analysis showed a high quercetin (QCE) content in the ICLEA extract. To prove the in vitro skin anticancer activity, an MTT assay was performed for the ICLEA extract in a malignant melanoma cell line, and the IC50 value was reported as 7.96 µg/mL. Conclusion I. coccinea leaf ethyl acetate extract revealed a significant total flavonoid content in analysis through the aluminum chloride method, and the presence of a high QCE content was confirmed by HPTLC analysis. The in vitro skin anticancer activity of ICLEA was confirmed by the MTT assay; therefore, it was concluded that the ICLEA extract was a potent fraction and was selected to develop a phytosome.

An Investigation into phytochemical constituents, antioxidant, antibacterial and anti-cataract activity of Alternanthera sessilis, a predominant wild leafy vegetable of South India

Author Index

Protective effects of Flacourtia indica aerial parts extracts against paracetamol-induced hepatotoxiciy in rats

Water, Ethanolic, Methanolic, Ethyl acetate, Chloroform, and Petroleum ether extracts from leaves of Artimesia sahariensis found in the Algerian Sahara, traditionally used for the treatment of fever, diarrhea, gut infection, and other infectious diseases, were tested in vitro for their antimicrobial activity against pathogenic bacteria and fungi. Antibacterial and antifungal activities were tested using agar disk diffusion and agar dilution method. All the extracted products showed antimicrobial activity against the tested strains. Petroleum ether and Ethyl acetate extracts of A. sahariensis leaves showed high inhibitory activity against the majority of strains tested, with minimal inhibitory concentration (MIC) ranging from 30–32 µg/ml. The largest inhibition zone was obtained with chloroform extract against Pseudomonas syringae pv. Tomato ATCC 1086 (18,33 ± 0,58 mm) and the MIC value of 40 µg/ml was obtained. The results showed that Methanolic and Ethyl acetate extracts have potent antifungal activity against Fusarium spp. CLMAS 11 with a MIC value of 80 µg/ml and an inhibition zone ranging from 17, 0 ± 1, 0 to 17, 33 ± 0, 58, respectively. This study reveals clearly that A. sahariensis can serve as a potential source of antimicrobial compounds.

Background Dracocephalum heterophyllum was a traditional Tibetan medicine possesses various pharmacological effects involved in anti-inflammatory, antibacterial activities. However, its anti-hepatitis, antioxidant activity and bioactive compounds have not been reported, the objective of this research work was to investigate the pharmacological activity and bioactive compounds of D. heterophyllum extracts.ResultsIn the present study, the anti-hepatics and antioxidant activities of four D. heterophyllum extracts (i.e. petroleum ether extracts, ethyl acetate extracts, n-BuOH extracts, and water extracts) were conducted. The main chemical constituent of petroleum ether and ethyl acetate extracts were also isolated using chromatographic techniques and identified by NMR spectroscopic methods. The anti-hepatitis assay showed that the petroleum ether and ethyl acetate extracts of D. heterophyllum significantly prolonged the mean survival times and reduced the mortality of mouse hepatitis model induced by concanavalin A (ConA). The levels of alanine transaminase, aspartate transaminase in blood serum could be decreased obviously by ethyl acetate extracts compared with ConA group (P < 0.01). The histological analysis demonstrated that the ethyl acetate extracts could inhibit apoptosis and necrosis caused by ConA. In addition, the antioxidant activities of the four extracts of D. heterophyllum were measured by DPPH assay, ABTS assay, anti-lipidperoxidation assay, ferric reducing antioxidant power assay, ferrous metal ions chelating assay and determination of total phenolic contents. The results showed that the ethyl acetate extract had the highest antioxidant activities, followed by petroleum ether extract. Finally, nine mainly compounds were isolated from the Petroleum ether and ethyl acetate extracts, including four triterpenes: oleanolic acid (1), ursolic acid (2), pomolic acid (3), 2α- hydroxyl ursolic acid (4), three flavonoids: apigenin-7-O-rutinoside (5), luteolin (8), diosmetin (9) and two phenolic acids: rosmarinic acid (6), methyl rosmarinate (7).ConclusionThe Ethyl acetate extract of D. heterophyllum had the highest anti-hepatitis and antioxidants activities, followed by petroleum ether extract. The bioactive substances may be triterpenes, flavonoids and phenolic acids, the ethyl acetate extracts of D. heterophyllum may be possible candidates in developing anti-hepatitis medicine.Electronic supplementary materialThe online version of this article (doi:10.1186/s40529-016-0133-y) contains supplementary material, which is available to authorized users.

Background:Breast cancer is the most common cause of deaths in women. The search for traditionally used medicinal plants which can serve as non-toxic and affordable anticancer drugs is the need of the hour. This study aimed to investigate the anticancer potential of extracts of L. coronopifolia against human breast carcinoma cell line (MDA-MB-321). Methods:The MDA-MB-231 cells were plated in 96 well plates and exposed to 10-1,000 µg/ml of L. coronopifolia for 24 h. The cytotoxic response of different extracts was measured by MTT assay, neutral red uptake (NRU) assay and cellular morphological alterations under the microscope.Results:A concentration-dependent decrease in the cell viability of MDA-MB-231 cells was observed after the exposure of petroleum ether, ethyl acetate, chloroform, and ethanol extracts of L. coronopifolia. The cell viability was found to be 82%, 89% and 98% at 1000, 500 and 250 µg/ml, respectively in petroleum ether, 37%, 75% and 88% at 1,000, 500 and 250 µg/ml, respectively in ethyl acetate extract, 30%, 35% and 64% at 1,000, 500 and 250 µg/ml, respectively in chloroform extract and 44%, 65% and 82% at 1000, 500 and 250 µg/ml, respectively in ethanolic extract of L. coronopifolia exposed MDA-MB-231 cells. The results also exhibited morphological alterations in MDA-MB-231 cells exposed to various extracts. The cells treated with 250- 1000 µg/ml lost their original morphology and cell linkage as compared to control cells. Conclusion:These preliminary results suggest the promising anticancer potential of petroleum ether, ethyl acetate, chloroform, and ethanol extracts of L. coronopifolia against MDA-MB-321 cells. Further studies are required to know the mechanism(s) involved in the cell death.

The high resistance evolution of protozoans to the existing antiparasitic drugs has necessitated the quest for novel and effective drugs against plasmodium and trypanosome parasites. As a result, this study aimed to assess the antiplasmodial and antitrypanosomal potentials of chloroform, ethyl acetate and ethanol leaf extracts of Oedera genistifolia. Standard biochemical procedures were explored for the plant extraction and gas chromatography-mass spectroscopy (GCMS) was used to identify the bioactive compounds in the crude extracts. The cytotoxic effects of the crude extracts were assessed against human cervix adenocarcinoma (HeLa cells) and their antiparasitic activities were investigated against Plasmodium falciparum strain 3D7 and Trypanosoma brucei brucei. GCMS analyses of the crude extracts revealed the bioactive compounds that could be responsible for the biological activities. The extracts had no cytotoxic effect on HeLa cells and demonstrated good antiplasmodial activity (chloroform extract: IC50 = 11.6 µg∙mL−1, ethyl acetate extract: IC50 = 3.3 µg∙mL−1 and ethanol extract: IC50 = 3.7 µg∙mL−1). Likewise, they showed excellent antitrypanosomal activity with IC50 = 0.5 µg∙mL−1 for chloroform and ethyl acetate extracts and IC50 = 0.4 µg∙mL−1 for the ethanol extract. Findings from the present study indicated that O. genistifolia could be a good source of strong antiplasmodial and antitrypanosomal agents.

The methanol, petroleum ether, chloroform, ethyl acetate, 1-butanol and water extracts were obtained by extraction of mountain germander (Teucrium montanum L). The total phenolic content in extracts was measured by Folin-Ciocalteu method. The 1-butanol extract had the highest phenolic content (296.00 mg/g). High performance liquid chromatography (HPLC) was employed to define qualitative and quantitative content of phenolic acids in mountain germander extracts. The largest number of phenolic acids were determined in ethyl acetate and 1-butanol extracts, while these acids were not present in petroleum ether extract. The highest content of phenolic acids (28.619 mg/g) had ethyl acetate extract and gentisic acid (14.432 mg/g) was its major component. Despite of a large number of phenolic acids in 1-butanol extract their content was only 3.740 mg/g.

The total phenols, flavonoids and carotenoids contents of Samadera indica methanol extracts of leaf, bark, seed, root and seed pod were determined. Among the methanol extracts of leaf, bark, seed, root and seed pod, the total phenols, flavonoids and carotenoids contents were found to be 105.67±0.243 mg catechol equivalent per gram for seed, 136.28±1.26 mg quercetin equivalent per gram for bark and 82.37 ±0.842 mg β-carotene equivalent per gram for seed which are higher than the activity of other extracts of the plant. The antioxidant activity of the petroleum ether, ethyl acetate, chloroform, methanol and aqueous extracts of leaf, bark, seed, root and seedpod of Samadera indica were established. The IC50value was found lower for bark methanol extracts 43.7±1.08 μg/ml for DPPH assay, seed methanol exhibited higher activity towards reducing power assay, lower IC50 value for seed methanol extracts 44.74±0.249 μg/ml for metal chelating assay, in phosphomolybdenum assay higher activity found for seed methanol extracts 199.08 mg/mlequ of ascorbic acid/100g of the plant extracts. Hydroxyl radical scavenging activity found to be higher for seed methanol extracts i.e. 78%. Significant activity towards antibacterial assay was exhibited by Samadera indica methanol extracts. 100% cytotoxicity observed by bark and leaf methanol extracts of the plant at 200 mg/ml dosage. These results confer the synergic effect of phenol, flavonoid and carotenoid contents in the plant extracts against degradative bioreactions. Further studies are needed to explore the potential compound from Samadera indica since which has high potential to be used in drug formulation.

Artocarpus altilis (breadfruit) leaf extracts in different solvent media (petroleum ether, methanol, and ethyl acetate) were assessed for antimicrobial activity. Breadfruit leaf extracts have been reported to have different phytoconstituents. The effect of leaf extracts in different solvent media on pathogenic organisms like Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus mutans, and Enterococcus faecalis was studied by disc diffusion assay and MIC (minimal inhibitory concentrations) values were investigated. Phytochemical compounds like steroids, phytosterols, gums and resins were found to be present in leaf extracts with different extraction media. Phenols and terpenoids were detected in ethyl acetate and methanol leaf extracts. Flavonoids were present in the petroleum ether and ethyl acetate leaf extracts, whereas tannins were detected only in the methanol leaf extract. Maximum zone of inhibition was observed for Strep. mutans, E. faecalis, S. aureus, and P. aeruginosa by using 50 μl of ethyl acetate and methanol leaf extracts, 20 μl of petroleum ether leaf extract, 25 μl of petroleum ether leaf extract, and 50 μl of methanol leaf extract, respectively. The MIC values were reported in between 0.3 and 0.6 mg/ml corresponding to variations in different solvent media used for leaf extracts against four different pathogenic bacteria.

Petroleum ether, ethyl acetate and methanolic leaf extracts of Viburnum toronis Killip et Smith were studied. From the petroleum ether extract, fatty acids of methyl esters such as methyl myristate, methyl palmitate, methyl stearate and methyl araquidonate were identified by means of GC-MS. From the ethyl acetate extract, 2-methylbutanoic, 2-methyl-2-butenoic, 3-methylbutanoic acids and 4-hydroxy-4-methylpentanone were identified by means of GC-MS. Through the isolated organ technique, uterine relaxant was verified; and through the method of writhings induced by acetic acid, the antinociceptive activity of methanolic extracts and the extract in ethyl acetate were verified. It was found that the extract in ethyl acetate showed the greatest in both uterine relaxant and antinociceptive activity at doses of 250 mg/kg.

A comparative study of in vitro antimicrobial, antioxidant and cytotoxic activity of Albizia lebbeck and Acacia nilotica stem bark

The aim of this study was to investigate the phytochemical, antioxidant and anticholinesterase activities of Iris suaveolens. After determining total phenolic and flavonoid contents of the petroleum ether, chloroform and methanol extracts prepared from the rhizomes, the antioxidant capacity of the extracts was established using β-carotene-linoleic acid and CUPRAC methods. The chloroform extract which was rich in phenolic content exhibited the highest inhibition of lipid peroxidation in the β-carotene-linoleic acid system, and the best cupric reducing antioxidant capacity among the tested extracts. The petroleum ether extract indicated moderate anticholinesterase activity while the chloroform extract revealed significant butyrylcholinesterase inhibitory activity (75.03 ± 1.29%). Spectroscopic methods were used for the structural elucidation of the compounds (1-13) isolated from the petroleum ether and chloroform extracts. Coniferaldehyde (6), having the highest antioxidant activity in the β-carotene-linoleic acid assay at 25 and 50 µg/mL, demonstrated also the best effect in the CUPRAC method among the tested compounds (1-12). 3-Hydroxyirisquinone (10) showed the best anticholinesterase activity among the tested compounds (1-4, 6-12), and coniferaldehyde exhibited almost the same butyrylcholinesterase inhibitory activity (82.60 ± 2.33%) as galantamine (86.26 ± 0.66%).