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1.  Phytochemical screening and free radical scavenging activity of Citrullus colocynthis seeds extracts 
Objective
To study the phytochemical screening of different extracts from Citrullus colocynthis (C. colocynthis ) seeds extracts and to assess their antioxidant activity on the DPPH free radical scavenging.
Methods
Phytochemical screening, total content of polyphenols and flavonoids of C. colocynthis seeds extracts, including a crude aqueous extract (E1), a defatted aqueous extract (E2), a hydromethanolic extract (HM), an ethyl acetate extract (EA) and a n-butanol extract (n-B) was carried out according to the standard methods and to assess their corresponding effect on the antioxidant activity of this plant.
Results
None of these extracts contained detectable amount of alkaloid, quinone, antraquinone, or reducing sugar. Catechic tannins and flavonoids were abundant in E1, HM and EA, whilst terpenoids were abundantly present in E1 and n-B but only weekly in HM. Coumarins were found in E2, EA and n-B. Polyphenols, expressed as gallic acid equivalent, amounted, per 100 g plant matter, to 329, 1002 and 150 mg in EA, HM an E1 respectively. Flavonoids, expressed as catechin equivalent, amounted, per 100 g plant matter to 620, 241 and 94 mg in EA, HM and E1 respectively. Comparable values were found in n-B and E1, with lower values in E2. Quercetin, myricetin and gallic acid were found in the EA and HM extracts by thin layer chromatography, The antioxidative effect of these extracts yielded, when tested at a concentration of 2 000 µg/mL in a 1,1-diphenyl-2-picrylhydrazyl assay, a reducing percentage of 88.8% with EA, 74.5% with HM and 66.2% with E1, and corresponding IC50 of 350, 580 and 500 µg/mL as compared to 1.1 µg/mL for ascorbic acid.
Conclusions
These qualitative and quantitative analytical data document the presence in C. colocynthis extracts of such chemical compounds as flavonoids responsible for the antioxidant activity, as well as other biological activities of this plant.
doi:10.1016/S2221-1691(13)60020-9
PMCID: PMC3609396  PMID: 23570014
Citrullus colocynthis; Polyphenols; Flavonoids; Free radical scavenging
2.  In vivo anti-ulcer, anti-stress, anti-allergic, and functional properties of Gymnemic Acid Isolated from Gymnema sylvestre R Br 
Background
Gymnema sylvestre is a highly valued ethno pharmacologically important medicinal plant used currently in many poly-herbal formulations due to its potential antidiabetic activity and other health benefits. The present study was carried out to analyze the anti-stress, anti-allergic, and antiulcer activity of the bioactive compounds present in Gymnema sylvestre leaves.
Methods
The preliminary phytochemical screening for bioactive compounds from aqueous extracts revealed the presence of alkaloids, triterpenes, flavonoids, steroids, and saponins. The antioxidant activities were investigated using DPPH radical scavenging method. The characterization of the extract was carried out using standard compound by High Performance Thin Layer Chromatography (HPTLC) and phytochemical analysis in terms of total phenol, total flavonoids, reducing power and antioxidant potentials, etc. The in vivo studies on albino mice proved the purified fraction has anti-stress/anti-allergic activity against milk induced leucocytosis/eosinophilia and able to inhibit the aspirin induced gastric ulcers.
Results
The quantitative estimation for aqueous extract exhibited total antioxidant (9.13 ± 0.04 μg/g), flavonoids (125.62 ± 26.84 μg/g), tannin (111.53 ± 15.13 μg/g), total phenol content (285.23 ± 1.11 μg/g) and free radical scavenging (52.14 ± 0.32%). Further the aqueous extract was consecutively purified by TLC and silica column chromatography. The purified fractions were characterized by HPTLC and GC-MS and the component was identified as gymnemic acid. The potency of the antimicrobial activity of the extract was studied with bacteria. Pharmacological experiments clearly demonstrated that the extracts of all plants given orally showed significant gastric protection against the asprin-induced gastric ulcer model in mice. Furthermore, healing effects were also confirmed through histopathological examination.
Conclusions
The aqueous extracts of the leaves of Gymnema sylvestre possess anti ulcerogenic, Anti allergic, Anti stress, properties that may be due to cytoprotective mechanism. These results support the ethno medical uses of the plant in the treatment of gastric ulcer.
doi:10.1186/1472-6882-14-70
PMCID: PMC3936851  PMID: 24559073
Gymnema sylvestre; Gymnemic acid; Anti- stress; Anti-ulcer; Anti- allergic; Antioxidant activities; Leaf extract
3.  Assessment of phytochemicals, antioxidant, anti-lipid peroxidation and anti-hemolytic activity of extract and various fractions of Maytenus royleanus leaves 
Background
Maytenus royleanus is traditionally used in gastro-intestinal disorders. The aim of this study was to evaluate the methanol extract of leaves and its derived fractions for various antioxidant assays and for its potential against lipid peroxidation and hemolytic activity.
Methods
Various parameters including scavenging of free-radicals (DPPH, ABTS, hydroxyl and superoxide radical), hydrogen peroxide scavenging, Fe3+ to Fe2+ reducing capacity, total antioxidant capacity, anti-lipid peroxidation and anti-hemolytic activity were investigated. Methanol extract and its derived fractions were also subjected for chemical constituents. LC-MS was also performed on the methanol extract.
Results
Qualitative analysis of methanol extract exhibited the presence of alkaloids, anthraquinones, cardiac glycosides, coumarins, flavonoids, saponins, phlobatannins, tannins and terpenoids. LC-MS chromatogram indicated the composition of diverse compounds including flavonoids, phenolics and phytoestrogens. Methanol extract, its ethyl acetate and n-butanol fractions constituted the highest amount of total phenolic and flavonoid contents and showed a strong correlation coefficient with the IC50 values for the scavenging of DPPH, hydrogen peroxide radicals, superoxide radicals, anti-lipid peroxidation and anti-hemolytic efficacy. Moreover, n-butanol fraction showed the highest scavenging activity for ABTS radicals and for reduction of Fe3+ to Fe2+.
Conclusions
Present results suggested the therapeutic potential of Maytenus royleanus leaves, in particular, methanol extract, ethyl acetate and n-butanol fraction as therapeutic agent against free-radical associated damages. The protective potential of the extract and or fraction may be attributed due to the high concentration of phenolic, flavonoid, tannins and terpenoids.
doi:10.1186/1472-6882-13-143
PMCID: PMC3702440  PMID: 23800043
Maytenus Royleanus; Antioxidant Activities; Phenolic Content; Solvent Extraction
4.  Evaluation of Abelmoschus moschatus extracts for antioxidant, free radical scavenging, antimicrobial and antiproliferative activities using in vitro assays 
Background
Abelmoschus moschatus Medik. leaves and seeds are considered as valuable traditional medicine. The aromatic seeds of this plant are aphrodisiac, ophthalmic, cardio tonic, antispasmodic and used in the treatment of intestinal complaints and check queasiness. To give a scientific basis for traditional usage of this medicinal plant, the seed and leaf extracts were evaluated for their antioxidant, free radical scavenging, antimicrobial and antiproliferative activities.
Methods
In this study, antioxidant, antimicrobial and antiproliferative activities of A. moschatus extracts were evaluated in a series of in vitro assay involving free radicals, reactive oxygen species and their IC50 values were also determined. The antioxidant activities of the seed and leaf extracts of A. moschatus were determined by total antioxidant, DPPH, and ferrous reducing antioxidant property (FRAP) methods. In addition, the antiproliferative activity was also evaluated using colorectal adenocarcinoma and retinoblastoma human cancer cell lines. Moreover, six bacterial reference strains, two gram-positive (Bacillus subtilis and Staphylococcus aureus), four gram-negative (Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Salmonella enterica paratyphi) and one fungal strain (Candida albicans) were used to evaluate its antimicrobial activity.
Results
The results from this study showed that the antioxidant activities of A. moschatus as determined by the total phenol, flavonoids, total antioxidant and FRAP methods were higher in leaf than that of the seed extracts. On the other hand, the aqueous overnight seed extract (AMS-I) has shown significant radical scavenging activity as in 1, 1- Diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide, hydroxyl radical, superoxide and lipid peroxidation as compared to other seed and leaf extracts. The AMS-I and AML-IV have shown activity against six and seven microorganisms respectively. Simulteneously, AMS-IV and AML-IV have demonstrated potential antiproliferative activity against two human cell lines - Colorectal adenocarcinoma (COLO-205) and retinoblastoma (Y79).
Conclusion
The seed and leaf extracts of A. moschatus possess significant antioxidant activity and could serve as free radical inhibitors or scavenger, or substitute, probably as primary antioxidants. The plant possesses moderate antibacterial activity against bacterial strains used in this study. Hydroalcoholic seed and leaf extracts also exhibited antiproliferative activity against two human cancer cell lines. A. moschatus may therefore, be a good candidate for functional foods as well as pharmaceutics.
doi:10.1186/1472-6882-11-64
PMCID: PMC3201038  PMID: 21849051
5.  Antioxidant and phytochemical properties of Carpobrotus edulis (L.) bolus leaf used for the management of common infections in HIV/AIDS patients in Eastern Cape Province 
Background
Carpobrotus edulis (Mesembryanthemaceae), also known as igcukuma in Xhosa language is a medicinal plant used by the traditional healers to treat common infections in HIV/AIDS patients. Based on this information, we researched on the plant phytoconstituents, as well as its inhibitory effect using aqueous and three different organic solvent extracts in order to justify its therapeutic usage.
Methods
Antioxidant activity of the extracts were investigated spectrophotometrically against 1,1- diphenyl-2-picrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) diammonium salt, hydrogen peroxide (H2O2), nitric oxide (NO), and ferric reducing power, Total phenols, flavonoids, flavonols, proanthocyanidins, tannins, alkaloids and saponins were also determined using the standard methods.
Results
Quantitative phytochemical analysis of the four solvent extracts revealed a high percentage of phenolics (55.7 ± 0.404%) in the acetone extract, with appreciable amount of proanthocyanidins (86.9 ± 0.005%) and alkaloids (4.5 ± 0.057%) in the aqueous extract, while tannin (48.9 ± 0.28%) and saponin (4.5 ± 0.262%) were major constituents of the ethanol extract. Flavonoids (0.12 ± 0.05%) and flavonols (0.12 ± 0.05%) were found at higher level in the hexane extract in comparison with the other extracts. The leaf extracts demonstrated strong hydrogen peroxide scavenging activity, with the exception of water and ethanol extracts. IC50 values of the aqueous and ethanolic extract against DPPH, ABTS, and NO were 0.018 and 0.016; 0.020 and 0.022; 0.05 and 0.023 mg/ml, respectively. The reducing power of the extract was found to be concentration dependent.
Conclusion
The inhibitory effect of the extracts on free radicals may justify the traditional use of this plant in the management of common diseases in HIV/AIDs patients in Eastern Cape Province. Overall, both aqueous and ethanol were found to be the best solvents for antioxidant activity in C. edulis leaves.
doi:10.1186/1472-6882-12-215
PMCID: PMC3528461  PMID: 23140206
Carpobrotus edulis; Solvent extraction; Antioxidant; Free radicals; Phytoconstituents
6.  In vitro antioxidant potential of dicliptera roxburghiana 
Background
Stress caused by free radicals accumulation result into many hazardous diseases. A number of investigations are focusing to find out the plant oriented natural antioxidant moieties. The basic aim of this research was to investigate the antioxidant potential, total Phenolic and flavonoids contents and photochemical screening of the crude methanol extract and its derived various fractions Dicliptera roxburghiana of Acanthaceae family.
Methods
Crude methanol extract of aerial parts of Dicliptera roxburghiana (DRME) was partitioned in to n-hexane (DRHF), chloroform (DRCF), ethyl acetate (DREF), n-butanol (DRBF) and the remaining soluble portion as residual aqueous fraction (DRAF). We evaluated the antioxidant activities of the extract and various fractions through different analytical methods such as DPPH, superoxide anion, ABTS, H2O2, hydroxyl radical and phosphomolybdate radical inhibition. In vitro lipid peroxidation and reducing power of the plant was also analyzed. Total flavonoid and phenolic contents of the extract and all fractions were also quantified. Plant was also subjected for preliminary phytochemical screening to confirm the presence or absence of various constituents in the plant.
Results
Phytochemical screening confirmed the presence of flavonoids, phenolics, tannins, alkaloids, saponins, terpenoids and coumarines. Quantitative analysis revealed the maximum amount of total phenolic and flavonoid contents in DRME while lowest in DRHF. Methanol extract, DREF, DRCF and DRBF exhibited promising antioxidant potential for DPPH, ABTS, H2O2, phosphomolybdate, superoxide anion and hydroxyl radical scavenging capabilities, while these were not appreciable for DRHF and DRAF. All fractions except DRHF and DRAF possess strong reducing power ability and showed appreciable lipid peroxidation inhibition.
Conclusion
These research investigations revealed that Dicliptera roxburghiana is a potent source of natural antioxidants. Hence the plant can be used for management of different stress and anxiety related ailments.
doi:10.1186/1472-6882-13-140
PMCID: PMC3717081  PMID: 23777321
Dicliptera roxburghiana; Lipid peroxidation; Total flavonoids; Antioxidants
7.  Antioxidant activity, total phenolic and total flavonoid contents of whole plant extracts Torilis leptophylla L 
Background
The aim of this study was to screen various solvent extracts of whole plant of Torilis leptophylla to display potent antioxidant activity in vitro and in vivo, total phenolic and flavonoid contents in order to find possible sources for future novel antioxidants in food and pharmaceutical formulations.
Material and methods
A detailed study was performed on the antioxidant activity of the methanol extract of whole plant of Torilis leptophylla (TLM) and its derived fractions {n-hexane (TLH), chloroform (TLC) ethyl acetate (TLE) n-butanol (TLB) and residual aqueous fraction (TLA)} by in vitro chemical analyses and carbon tetrachloride (CCl4) induced hepatic injuries (lipid peroxidation and glutathione contents) in male Sprague-Dawley rat. The total yield, total phenolic (TPC) and total flavonoid contents (TFC) of all the fractions were also determined. TLM was also subjected to preliminary phytochemical screening test for various constituents.
Results
The total phenolic contents (TPC) (121.9±3.1 mg GAE/g extract) of TLM while total flavonoid contents (TFC) of TLE (60.9 ±2.2 mg RTE/g extract) were found significantly higher as compared to other solvent fractions. Phytochemical screening of TLM revealed the presence of alkaloids, anthraquinones, cardiac glycosides, coumarins, flavonoids, saponins, phlobatannins, tannins and terpenoids. The EC50 values based on the DPPH (41.0±1 μg/ml), ABTS (10.0±0.9 μg/ml) and phosphomolybdate (10.7±2 μg/ml) for TLB, hydroxyl radicals (8.0±1 μg/ml) for TLC, superoxide radicals (57.0±0.3 μg/ml) for TLM and hydrogen peroxide radicals (68.0±2 μg/ml) for TLE were generally lower showing potential antioxidant properties. A significant but marginal positive correlation was found between TPC and EC50 values for DPPH, hydroxyl, phosphomolybdate and ABTS, whereas another weak and positive correlation was determined between TFC and EC50 values for superoxide anion and hydroxyl radicals. Results of in vivo experiment revealed that administration of CCl4 caused a significant increase in lipid peroxidation (TBARS) while decrease in GSH contents of liver. In contrast, TLM (200 mg/kg bw) and silymarin (50 mg/kg bw) co-treatment effectively prevented these alterations and maintained the antioxidant status.
Conclusion
Data from present results revealed that Torilis leptophylla act as an antioxidant agent due to its free radical scavenging and cytoprotective activity.
doi:10.1186/1472-6882-12-221
PMCID: PMC3524761  PMID: 23153304
Torilis leptophylla; DPPH; Antioxidant; Oxidative stress; TBARS
8.  Polyphenol Content and Antioxidant Properties of Colored Soybean Seeds from Central Europe 
Journal of Medicinal Food  2012;15(1):89-95.
Abstract
The antioxidant activity and contents of various polyphenol classes in the seeds of seven soybean varieties of different seed color and one yellow seed cultivar, representing a reference genotype, were evaluated. Total polyphenols and tannins were determined after extraction of plant material with 70% aqueous acetone, and total flavonoids were extracted with methanol and acetic acid, whereas anthocyanins were extracted with 20% aqueous ethanol. In addition, isoflavone content and composition were determined using high-performance liquid chromatography analysis. Antioxidant activity of seed extracts was evaluated by the 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity assay. A positive linear correlation between antioxidant activity and contents of total polyphenols and anthocyanins was established. The highest antioxidant activity was observed in the extracts of black and brown varieties, which also showed high levels of all polyphenol classes examined. Yellow seed had the highest total isoflavone content (3.62 mg/g of dry material). The highest concentration of total daidzein was determined in black seeds (>2.0 mg/g of dry material), and the highest total glycitein and genistein contents occurred in the yellow cultivar (0.53 and 1.49 mg/g of dry material, respectively). According to our results, varieties of black and brown seeds could be of special interest not only for their large content of total polyphenols, ranging from 4.94 to 6.22 mg of gallic acid equivalents/g of dry material, but also for their high content of natural antioxidants such as anthocyanins.
doi:10.1089/jmf.2010.0329
PMCID: PMC3249631  PMID: 21861721
anthocyanins; 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity; flavonoids; isoflavones; tannins
9.  Antioxidant activities of ethanol extracts and fractions of Crescentia cujete leaves and stem bark and the involvement of phenolic compounds 
Background
Antioxidant compounds like phenols and flavonoids scavenge free radicals and thus inhibit the oxidative mechanisms that lead to control degenerative and other diseases. The aim of this study was to investigate the antioxidant activity in vitro, total phenolic and flavonoid contents in ethanol extracts and fractions of Crescentia cujete leaves and stem bark.
Methods
Crescentia cujete leaves and bark crude ethanol extract (CEE) and their partitionates petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF) and aqueous (AQF) were firstly prepared. Different established testing methods, such as 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical, ferric reducing power (FRP), and total antioxidant capacity (TAC) assays were used to detect the antioxidant activity. Further, the total yield, total phenolic (TPC) and total flavonoid contents (TFC) of CEE and all the fractions were determined. Ethanol extracts of both leaves and stem bark were also subjected to preliminary phytochemical screening to detect the presence of secondary metabolites, using standard phytochemical methods (Thin layer chromatography and spray reagents).
Results
Phytochemical screening of crude ethanol extract of both leaves and stem bark revealed the presence of steroids, flavonoids, saponins, tannins, glycosides and terpenoids. All the fractions and CEE of leaves and bark exhibited antioxidant activities, however, EAF of leaves showing the highest antioxidant activity based on the results of DPPH, FRP and TAC assay tests. The above fraction has shown the significant DPPH scavenging activity (IC50 = 8.78 μg/ml) when compared with standard ascorbic acid (IC50 =7.68 μg/ml). The TAC and FRP activities increased with increasing crude extract/fractions content. The TPC (371.23 ± 15.77 mg GAE/g extract) and TFC (144.64 ± 5.82 mg QE/g extract) of EAF of leaves were found significantly higher as compared to other solvent fractions for both leaves and bark. TPC were highly correlated with the antioxidant activity (R2 = 0.9268 and 0.8515 in DPPH test for leaves and bark, respectively).
Conclusion
The results of the study show that leaves of C. cujete possesses significant free radical scavenging properties compared with stem bark and a clear correlation exists between the antioxidant activity and phenolic content.
doi:10.1186/1472-6882-14-45
PMCID: PMC3937116  PMID: 24495381
Calabash tree; Oxidative stress; Crude extracts; Free radicals; Anti-aging
10.  Phytochemical constituents and antioxidant activities of the whole leaf extract of Aloe ferox Mill 
Pharmacognosy Magazine  2011;7(28):325-333.
Background:
Aloe ferox Mill. (Asphodelaceae) is used in South Africa for the treatment of constipation among various ailments. Despite the extensive studies conducted on the antioxidant activities of the leaf gel and pulp extract of the plant, there is no information on the antioxidant properties of the whole leaf extract of the species.
Materials and Methods:
The antioxidant activities of ethanol, acetone, methanol and aqueous extracts of A. ferox were investigated spectrophotometrically against 1,1- diphenyl-2-picrylhydrazyl (DPPH), 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) diammonium salt, hydrogen peroxide (H2O2), nitric oxide (NO), lipid peroxidation and ferric reducing power. Total phenols, flavonoids, flavonols, proanthocyanidins, tannins, alkaloids and saponins were also determined using the standard methods.
Results:
The percentage compositions of phenols (70.33), flavonols (35.2), proanthocyanidins (171.06) and alkaloids (60.9) were significantly high in the acetone extract, followed by the ethanol extract with values of 70.24, 12.53, 76.7 and 23.76 respectively, while the least composition was found in the aqueous extract. Moreover, both flavonoids and saponins contents were appreciably high in both methanol and ethanol extracts, while others were very low. Tannins levels were, however, not significantly different (P > 0.05) in all the solvent extracts. At 0.5 mg/ml, the free radical scavenging activity of the methanol, acetone and ethanol extracts showed higher inhibition against ABTS, hydrogen peroxide and nitric oxide radicals. Whereas, scavenging activity of the extracts against DPPH* and lipid peroxidation were observed at a concentration of 0.016 and 0.118 mg/ml respectively in comparison to the butylated hydroxyltoluene (BHT), gallic acid and rutin. The ferric reducing potential of the extracts was concentration dependent and significantly different from that of vitamin C and BHT.
Conclusion:
The present study showed high level of radical scavenging activity by ethanol and methanol whole leaf extracts of A. ferox with higher antioxidant activities than acetone and aqueous extracts. The significant differences show that the whole leaf extract could be used as a potent antioxidant in medicine and food industries.
doi:10.4103/0973-1296.90414
PMCID: PMC3261067  PMID: 22262936
Aloe ferox; antioxidant; asphodelaceae; free radicals; phytochemicals
11.  Biological activity and microscopic characterization of Lythrum salicaria L 
Background
There are several plants have been used worldwide in the folk medicine with high incidence for treatment of human disorders, of which Lythrum salicaria belongs to the Lythraceae family has traditionally reputation for some medicinal usage and recently many biological and pharmacological activity of the plant have been studied.
Methods
In this study, microscopic characterizations of the aerial parts of the plant were determined. Moreover, the plant extract (aqueous methanol 80%) was subjected to an anti-diabetic activity test (in a rat model of streptozocin induced diabetes), anti-Helicobacter pylori (using disc diffusion method) along with antioxidant activity against DPPH (stable free radical) tests. Besides, total flavonoids, phenols, tannins, as well as polysaccharides contents have been assessed using spectroscopic methods.
Results
The microscopic properties of the plant fragments revealed anomocytic stomata, conical shape trichomes, and abundant spherical pollen grains as a characteristic pattern for the aerial parts of the plant. The extract of the plant at concentration of 15 g/kg showed mild lowering activity on blood glucose level to 12.6% and 7.3% after 2 and 3 h of administration. Additionally, clinically isolated H. pylori strain was inhibited with the plant extract at concentration of 500 mg/mL (zone of inhibition: 17 ± 0.08 mm). Moreover, IC50 values for DPPH inhibition of the plant extract, vitamin E, BHA were examined as 13.5, 14.2, and 7.8 μg/mL, respectively. Total flavonoids, phenols, tannin, and polysaccharides contents of the extract were successfully evaluated as 5.8 ± 0.4 μg QE/mg EXT, 331 ± 3.7 μg GAE/mg EXT, 340 ± 2.3 μg TAE/mg EXT, 21 ± 0.2 μg GE/mg EXT, respectively.
Conclusions
The results suggested that L. salicaria has low anti-diabetic and anti-Helicobacter pylori effects, but high antioxidant activity, just the same as positive standard (vitamin E), which might be attributed to the high content of phenolic compounds in the extract.
doi:10.1186/2008-2231-21-61
PMCID: PMC3751246  PMID: 23885663
Lythrum salicaria; Antioxidant; Diabetes; H. pylori; Microscopy characterization
12.  Phytochemical analysis and in vitro antioxidant acitivity of hydroalcoholic seed extract of Nymphaea nouchali Burm. f. 
Objective
To evaluate the phytochemical constituents and the antioxidant activity of hydroalcoholic extract of Nymphaea nouchali seed locally prescribed as a diet for diabetes mellitus.
Methods
The antioxidant and free radical scavenging activity of hydroalcoholic extract of the plant was assessed against 1,1 diphenyl-2-picryl hydrazyl (DPPH), nitric oxide and lipid peroxidation using standard protocols. Total phenolics, flavonoids and tannins were also determined.
Results
Phytochemical analysis revealed the presence of phenols, flavones, tannins, protein, reducing sugars, glycosides, saponins, alkaloids and steroids. The activities of plant extract against DPPH, nitric oxide and lipid peroxidation was concentration dependent with IC50 value of 42.82, 23.58 and 54.65 µg/mL respectively. The total antioxidant capacity was high with 577.73 mg vitamin E/g of the extract and showed a moderately high vitamin C content of 197.22 mg/g. The total tannin content of hydroalcoholic seed extract was high (195.84 GE/g), followed by phenolics (179.56 GE/g) and flavonoids (23.55 QE/g).
Conclusion
Our findings provide evidence that the crude extract of Nymphaea nouchali is a potential source of natural antioxidants and this justifies its use in folkloric medicine.
doi:10.1016/S2221-1691(13)60174-4
PMCID: PMC3793162
Nymphaea stellata; Phytochemical; Antioxidant; Scavenging; Free radicals; DPPH; Nitric oxide; Lipid peroxidation; Phenols; Flavonoids
13.  Zizyphus lotus L. (Desf.) modulates antioxidant activity and human T-cell proliferation 
Background
Zizyphus lotus L. (Desf.) also known as Jujube, is a deciduous shrub which belongs to Rhamnaceae family. This plant is used in Algerian traditional medicine for its anti-diabetic, sedative, analgesic, anti-inflammatory and hypoglycaemic activities. In the present study, we determined the concentrations of different vitamins (vitamin A, C and E) and fatty acids in root, stem, leaves, fruit pulp and seed of Zizyphus lotus L. (Desf.) and assessed the effects of their aqueous extracts on antioxidant status and human T-cell proliferation.
Methods
Aqueous filtrates from different parts, i.e, root, leaf, stem, fruit pulp and seed, of Zizyphus lotus L. (Desf.) were prepared. Vitamin C levels were determined by precipitating with 10% trichloroacetic acid and vitamin A and E were assessed by HPLC. Lipid composition of these extracts was determined by gas-liquid chromatography. Anti-oxidant capacity was evaluated by using anti-radical resistance kit [Kit Radicaux Libres (KRL@; Kirial International SA, Couternon, France)]. T-cell blastogenesis was assessed by the incorporation of 3H-thymidine. IL-2 gene expression was evaluated by RT-qPCR.
Results
Our results show that fruit pulp contained higher vitamin A and C contents than other parts of the plant. Furthermore, the fruit pulp was the richest source of linoleic acid (18:2n-6), a precursor of n-6 fatty acids. Fruit seeds possessed higher vitamin C levels than leaves, roots and stem. The leaves were the richest source of vitamin E and linolenic acid (18:3n-3), a precursor of n-3 fatty acids. The antioxidant capacity of the different extracts, measured by KRL@ test, was as follows: pulp < seed
Conclusion
Seed extracts exerted the most potent immunosuppressive effects on T cell proliferation and IL-2 mRNA expression. The results of the present study are discussed in the light of their use to modulate the immune-mediated diseases.
doi:10.1186/1472-6882-10-54
PMCID: PMC2955679  PMID: 20868496
Pharmacognosy Research  2010;2(3):152-158.
Background:
Leaves of Tephrosia purpurea Linn. (sarpankh), belonging to the family Leguminaceae, are used for the treatment of jaundice and are also claimed to be effective in many other diseases. This research work was undertaken to investigate the in vitro antioxidant activity of aqueous and ethanolic extracts of the leaves.
Method:
The therapeutic effects of tannins and flavonoids can be largely attributed to their antioxidant properties. So, the quantitative determinations were undertaken. All the methods are based on UV-spectrophotometric determination.
Result:
The total phenolic content of aqueous and ethanolic extracts showed the content values of 9.44 ± 0.22% w/w and 18.44 ± 0.13% w/w, respectively, and total flavonoid estimation of aqueous and ethanolic extracts showed the content values of 0.91 ± 0.08% w/w and 1.56 ± 0.12%w/w, respectively, for quercetin and 1.85 ± 0.08% w/w and 2.54 ± 0.12% w/w, respectively, for rutin. Further investigations were carried out for in vitro antioxidant activity and radical scavenging activity by calculating its percentage inhibition by means of IC50values, all the extracts’ concentrations were adjusted to fall under the linearity range and here many reference standards like tannic acid, gallic acid, quercetin, ascorbic acid were taken for the method suitability.
Conclusion:
The results revealed that leaves of this plant have antioxidant potential. The results also show the ethanolic extract to be more potent than the aqueous decoction which is claimed traditionally. In conclusion, T. purpurea Linn. (Leguminosae) leaves possess the antioxidant substance which may be responsible for the treatment of jaundice and other oxidative stress-related diseases.
doi:10.4103/0974-8490.65509
PMCID: PMC3141306  PMID: 21808558
Ferric reducing antioxidant power; 1, 1-diphenyl-2-picrylhydrazyl-radical scavenging assay; Tephrosia purpurea Linn. (Leguminosae); total phenolic content
Background
Many oxidative stress related diseases are as a result of accumulation of free radicals in the body. A lot of researches are going on worldwide directed towards finding natural antioxidants of plants origins. The aims of this study were to evaluate in vitro antioxidant activities and to screen for phytochemical constituents of Helichrysum longifolium DC. [Family Asteraceae] aqueous crude extract.
Methods
We assessed the antioxidant potential and phytochemical constituents of crude aqueous extract of Helichrysum longifolium using tests involving inhibition of superoxide anions, DPPH, H2O2, NO and ABTS. The flavonoid, proanthocyanidin and phenolic contents of the extract were also determined using standard phytochemical reaction methods.
Results
Phytochemical analyses revealed the presence of tannins, flavonoids, steroids and saponins. The total phenolic content of the aqueous leaf extract was 0.499 mg gallic acid equivalent/g of extract powder. The total flavonoid and proanthocyanidin contents of the plant were 0.705 and 0.005 mg gallic acid equivalent/g of extract powder respectively. The percentage inhibition of lipid peroxide at the initial stage of oxidation showed antioxidant activity of 87% compared to those of BHT (84.6%) and gallic acid (96%). Also, the percentage inhibition of malondialdehyde by the extract showed percentage inhibition of 78% comparable to those of BHT (72.24%) and Gallic (94.82%).
Conclusions
Our findings provide evidence that the crude aqueous extract of H. longifolium is a potential source of natural antioxidants, and this justified its uses in folkloric medicines.
doi:10.1186/1472-6882-10-21
PMCID: PMC2877649  PMID: 20470421
ISRN Pharmacology  2013;2013:691372.
The present study reports the antioxidant and membrane protective activities of Calotropis procera aqueous root extract using several in vitro assays along with the determination of phenolic as well as flavonoid contents. Total phenol and flavonoid contents in extract were 15.67 ± 1.52 mg propyl gallate equivalent/g and 1.62 ± 0.05 mg quercetin equivalent/g, respectively. UV-visual spectroscopic scanning of the extract indicated the presence of glycoside-linked tannins or flavonoids. The extract exhibited appreciable reducing power signifying hydrogen donating potential. DPPH radical scavenging assay revealed substantial free radical scavenging activity (42–90%) in the extracts. Concentration dependent response was observed in the metal ion chelating activity (16–95%). Extracts also provided protection against iron induced lipid peroxidation in rat tissue (liver, brain, and kidney) homogenates. Comparatively better protective efficacy against peroxidative damage was observed in liver (71%) followed by kidney (65%) and brain (60%) tissues. Positive correlation (r2 = 0.756) was observed between DPPH free radical scavenging activity and reducing power of extract. Similarly strong positive correlation (r2 ≈ 0.756) was observed between metal ion chelating ability and percentage lipid peroxidation inhibition in different tissues. The study demonstrated considerable protective efficacy in C. procera root aqueous extracts against free radical and metal ion mediated oxidative damage.
doi:10.1155/2013/691372
PMCID: PMC3809601  PMID: 24222863
BMC Research Notes  2013;6:121.
Background
Research on natural products has gained a wide popularity due to the potential of discovering active compounds. The antioxidant properties contained in plants have been proposed as one of the mechanisms for the observed beneficial effect. Therefore, the present study investigated the antioxidant activity and total phenolic contents of various solvent extracts of Albizia procera leaves.
Methods
Antioxidant activity of the methanol extract and its derived fractions petroleum ether (APP), carbon tetrachloride (APC), dichloromethane (APD), ethyl acetate (APE), and residual aqueous fraction (APA) of the leaves of Albizia procera was performed by in vitro chemical analyses. Total phenolic content of the APM and other five fractions were also determined. APM and its derived fractions were also subjected to preliminary phytochemical screening test for various constituents.
Results
Phytochemical screening revealed the presence of saponins, steroids, tannins, glycosides and flavonoids in the extracts. Amongst the extracts, APE showed the highest total phenolic content (449.18 ± 18.41mg of gallic acid equivalent/g of extract). In DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging test, the IC50 value of APM, APP, APC, APD, APE and APA was 43.43, 63.60, 166.18, 41.15, 11.79, and 63.06 μg/mL, respectively. Therefore, among the APM and its derived fractions, APE showed the highest antioxidant activity which is comparable to that of standard ascorbic acid (AA) (IC50 10.12 μg/mL). The total antioxidant capacity was found to be varied in different fractions. The reducing activity on ferrous ion was ranked as APE > APD > APM > APA > APC.
Conclusion
The above evidences suggest that APE of A. procera leaf is a potential source of natural antioxidant and can be used to prevent diseases associated with free radicals.
doi:10.1186/1756-0500-6-121
PMCID: PMC3621163  PMID: 23531304
Antioxidants; Free radical scavenging; Phytochemical constituents; Total phenolic content
Objective
To evaluate the phytochemical constituents and antioxidant activities of aqueous extract of Schotia latifolia (S. latifolia) bark locally used for the treatment of oxidative stress-induced ailments in South Africa.
Methods
The antioxidant and free radical scavenging activity of aqueous extract of the plant was assessed against 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide (NO), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and the ferric reducing agent. Total phenolics, flavonoids, flavonols and proanthocyanidins were also determined to assess their corresponding effect on the antioxidant activity of this plant.
Results
The activities of plant extract against DPPH, ABTS and NO radicals were concentration dependent with IC50 value of 0.06, 0.05 and 0.05 mg/mL, respectively. The reducing power of the extract was greater than that of butylated hydroxyl toluene (BHT) and ascorbic acid which were used as standard drugs in a concentration dependent manner. The total phenolics content of the aqueous bark extract was (193.33±0.03 TE/g), followed by flavonoids (72.70±0.01 QE/g), proanthocyanidins (48.76±0.00 CE/g) and flavonols (47.76±0.21 QE/g). Phytochemical analysis revealed the presence of percentage tannin (11.40±0.02), alkaloid (9.80±0.01), steroids (18.20±0.01), glycosides (29.80±0.01) and saponins (6.80±0.00). The results exhibited a positive linear correlation between these polyphenols and the free radical scavenging activities.
Conclusions
Our findings provide evidence that the crude aqueous extract of S. latifolia is a potential source of natural antioxidants and this justifies its uses in folkloric medicines.
doi:10.1016/S2221-1691(11)60204-9
PMCID: PMC3609248  PMID: 23569880
Schotia latifolia; Free radicals; Phytochemical; Antioxidant activity; Oxidative stress; Natural antioxidant; Reducing power; Phenolics; Flavonoids; Scavenging activity; Polyphenol
Objective
To analyse qualitative and quantitative phytochemical and evaluate in vitro antioxidant properties of various alcoholic and aqueous extracts of leaf and root parts of Hypochaeris radicata.
Methods
Preliminary phytochemical analysis for alkaloids, cardiac glycosides, flavonoids, glycosides, phenols, resins, saponins, steroids, tannins, terpenoids and triterpenoids and quantitative phytochemical analysis for alkaloids, total phenolics, total flavonoids, tannins, saponins and ascorbic acid were made by following standard procedures. In vitro antioxidant properties were evaluated by assessing DPPH•, NO• and ABTS•+, radical scavenging abilities and assaying the reducing power, β-carotene and antihemolytic activities by adapting standard methods.
Results
The quantitative phytochemical analysis of this species exhibited the presence of alkaloids, total phenolics, total flavonoids, tannins, saponins and ascorbic acid in considerable quantity. The in vitro antioxidant activity of the species, Hypochaeris radicata clearly demonstrated that both the leaf and root parts have prominent antioxidant properties.
Conclusions
From this study, it can be concluded that the species is effective in scavenging free radicals and has the potential to be a powerful antioxidant.
doi:10.12980/APJTB.4.2014C1030
PMCID: PMC4025295  PMID: 25183112
Hypochaeris radicata; Phytochemical analysis; In vitro antioxidant activities
The side effects of synthetic antioxidants have been considered in different studies. Accordingly, there is an increasing interest toward the use of natural substances instead of the synthetic ones. In this study, the aqueous and ethanolic extracts of Pistacia vera leaves and fruits as well as hydroalcoholic extract of gum were tested for a possible antioxidant activity using in vitro methods. Deoxyribose assay, erythrocyte membrane lipid peroxidation and liver misrosomal non- enzymatic lipid peroxidation tests were used as an in-vitro model for determination antioxidant activity. The extract were evaluated at different concentratios: 25,100, 250, 500 and 1000 μg/mL. In all procedures, all extracts showed free radical scavenging activity. The effect of ethanolic extract of P. vera fruit at 1000 μg/mL was quite similar to positive control (DMSO 20 mM) in deoxyribose method. In two other tests, the ethanolic extracts of fruits and leaves were more effective than the aqueous extracts to inhibit malondialdehyde generation. Phytochemical tests showed the presence of flavonoids and tannins in Pistocia vera extracts. The present study showed that extracts of different part of P. vera have antioxidant activity in different in vitro methods. The ethanolic extracts of leaves and fruits showed more roles for antioxidant properties and gum hydroalcoholic extract demonstrated less antioxidant effect.
PMCID: PMC3813125  PMID: 24250515
Pistacia vera; Pistachio; Antioxidant; Deoxyribose; Free radical; Lipid peroxidation
Background
In South Africa, Calpurnia aurea (Ait.) Benth is used to destroy lice and to relieve itches, to destroy maggots and to treat allergic rashes, particularly those caused by caterpillars. Antioxidants play an important role protecting against damage by reactive oxygen species. Plants containing flavonoids have been reported to possess strong antioxidant properties.
Methods
The antibacterial, antioxidant activities and phenolic contents of the methanol extracts of the leaves and stems of Calpurnia aurea were evaluated using in vitro standard methods. Spectrophotometry was the basis for the determinations of total phenol, total flavonoids, flavonols, and proanthocyanidins. Tannins, quercetin and catechin equivalents were used for these parameters. The antioxidant activities of the stem extract of Calpurnia aurea were determined by ABTS, DPPH, and ferrous reducing antioxidant property (FRAP) methods. Laboratory isolates of 10 bacteria species which included five Gram-positive and five Gram-negative strains were used to assay for antibacterial activity of this plant.
Results
The results from this study showed that the antioxidant activities of the stem extract of Calpurnia aurea as determined by the total phenol, flavonoids, and FRAP methods were higher than that of the leaves. On the other hand, the leaf extract of the plant has higher level of total flavonols and proanthocyanidins. The leaf extract also has higher radical scavenging activity as shown in 1, 1-Diphenyl-2-picrylhydrazyl (DPPH), and 2,2¿-azinobis-3- ethylbenzothiazoline-6-sulfonic acid (ABTS) assay. The leaf extract showed activity against seven of the bacterial organisms.
Conclusion
The results from this study indicate that the leaves and stem extracts of Calpurnia aurea possess antioxidant properties and could serve as free radical inhibitors or scavenger or, acting possibly as primary antioxidants. Although, the antibacterial properties of Calpurnia aurea are not as effective as the standard drugs- Chloramphenicol and Streptomycin, they still possess some activity against bacterial strains used in this study. Calpurnia aurea may therefore be a good candidate for functional foods as well as pharmaceutical plant-based products.
doi:10.1186/1472-6882-8-53
PMCID: PMC2556645  PMID: 18803865
Background
Roots and leaves of the Cermela Hutan (Phyllanthus gomphocarpus Hook. F) plant were studied to determine antioxidant activity, phytochemical compounds, proportion of carbohydrate, crude protein, moisture, ash, fat, total phenolic content (TPC), and total flavonoid content (TFC).
Material/Methods
Ten percent (10%) aqueous extract from both Phyllanthus gomphocarpus roots (PGR) and leaves (PGL) were used in this study. Antioxidant activity characterization by TPC, TFC, Ferric Reducing Antioxidant Power (FRAP), 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, and phytochemical screening, as well as proximate analysis from both extracts were analyzed in this study.
Results
Phyllanthus gomphocarpus roots (PGR) and leaves (PGL) tested positive for flavonoid, saponin, tannins, and terpenoids, but PGR showed negative result for anthraquinones. In average weight of 100.0 g dry sample, the carbohydrates, protein, moisture, ash, fat, and energy content in PGR and PGL were 80.9%, 5.5%, 7.8%, 3.4%, 2.4%, and 367 Kcal/100 g, and 66.5%, 14.8%, 10.7%, 6.5%, 1.5%, and 399 Kcal/100g, respectively. Antioxidant assessments using FRAP and DPPH assay showed that PGL extracts possessed higher antioxidant capacity by reducing the ferric ion-TPTZ complex by 0.14 mg/ml ±0.0018 and higher scavenging activity, 83.83% ±0.54 as compared to PGR, 0.07 mg/ml ±0.0035 for FRAP and 62.87% ±1.33 for DPPH, respectively. The total phenolics content was significantly higher in PGL (208.77 mg GAE/g ±3.79) as compared to PGR (27.53 mg GAE/g ±0.42). However, there was no significant different in the total flavonoid contents for PGR (34.8 mg QE/g ±3.12) and PGL (32.43 mg QE/g ±3.92).
Conclusions
Further investigations are suggested to isolate and characterize the other active constituents from this plant in combatting diseases.
doi:10.12659/MSMBR.892345
PMCID: PMC4237070  PMID: 25381551
Antioxidants; Phyllanthus gomphocarpus; Phytochemicals; Proximate analysis
Background
The use of traditional medicine at the primary health care level is widespread and plant-based treatments are being recommended for curing various diseases by traditional medical practitioners all over the world. The phytochemicals present in the fruits, vegetables and medicinal plants are getting attention day-by-day for their active role in the prevention of several human diseases. Abrus precatorius is a widely distributed tropical medicinal plant with several therapeutic properties. Therefore in the present study, A. precatorius leaf extracts were examined for their antioxidant and cytotoxic properties in vitro in order to discover resources for new lead structures or to improve the traditional medicine.
Methods
In this study, antioxidant and antiproliferative properties of the different leaf extracts (hexane, ethyl acetate, ethanol and water) from A. precatorius were investigated along with the quantification of the polyphenol and flavonoid contents. The ability of deactivating free radicals was extensively investigated with in vitro biochemical methods like DPPH•, •OH, NO, SO2- scavenging assays and inhibition capability of Fe(II)-induced lipid peroxidation. Furthermore, antiproliferative activities using different human cancer cell lines and primary cell line was carried out by MTT method.
Results
Total phenolic content and total flavonoid content of the extracts were found in the range of 1.65 ± 0.22 to 25.48 ± 0.62 GAE mg/g dw and 6.20 ± 0.41 to 17.16 ± 1.04 QE mg/g dw respectively. The experimental results further revealed that A. precatorius extracts showed strong antiradical properties, capable to chelate Fe2+ and possess good inhibition ability of lipid peroxidation. In addition, as a first step towards the identification of phytoconstituents endowed with potent chemopreventive activities, we evaluated the inhibitory effects of A. precatorius extracts on the proliferation of four different human tumour cell lines such as human colon adenocarcinoma cells (Colo-205), human retinoblastoma cancer cells (Y79), human hepatocellular carcinoma cells (HepG2) and Leukemia cells (SupT1). Ethanol extract (APA) and ethyl acetate extract (APE) of A. precatorius had apparent capabilities of inhibiting the survival of tested human cancer cell lines. Moreover, it was observed that the A. precatorius extracts did not inhibit the growth of mice peritoneal macrophages, thus confirming that plants extracts are selective against the cancer cell lines.
Conclusion
This work provides a scientific support for the high antioxidant and antiproliferative activity of this plant and thus it may find potential applications in the treatment of the diseases caused by ROS. Further studies are needed to confirm in vivo anti-tumorgenicity and subsequent chemical characterization of the active molecule(s).
doi:10.1186/1472-6882-13-53
PMCID: PMC3600005  PMID: 23452983
Objective
To assess antioxidant activities of different aerial parts of Thymelaea hirsuta (T. hirsuta) from west Algeria, and to search for new sources of safe and inexpensive antioxidants.
Methods
Samples of leaves, stems and flowers from T. hirsuta were tested for total phenolic content, flavonoids content, and evaluation its total antioxidant activity, were done using the spectrophotometric analyses.
Results
Results of preliminary phytochemical screening of leaf, flower and stem of T. hirsuta revealed the presence of tannins, alkaloids, steroids, saponins, coumarins, reducteurs compound and anthraquinones. The total phenolics and flavonoids were estimated. The aqueous extracts of the aerial parts of T. hirsuta showed potent in vitro antioxydant activities using various models viz, DPPH scavenging assay, ferric reducing antioxidant power (FRAP) and ABTS radical scavenging activity.
Conclusions
On the basis of the results obtained, T. hirsuta extracts are rich sources of natural antioxidants appears to be an alternative to synthetic antioxidants and this justifies its therapeutic usage.
doi:10.1016/S2222-1808(14)60324-8
PMCID: PMC4032046
Phytochemicals; Flavonoids; Total phenolics; Antioxidant; Thymelaea hirsute
Background
Buddleja saligna Willd (Loganiaceae) is a small to medium-sized evergreen tree; trunk short, often gnarled and crooked; crown dense, rounded or domed-shaped; foliage greyish green. The wild olives are traditionally used to lower blood pressures in many parts of the world. In southern Africa, bark and leaf decoctions are used to treat colic, coughs, colds, sore eyes, urinary problems and as purgatives.
Methods
The antibacterial, antioxidant activities and phenolic contents of the methanol extracts of the leaves and stems of Buddleja saligna were evaluated using in vitro standard methods. Spectrophotometry was the basis for the determinations of total phenol, total flavonoids, flavonols, and proanthocyanidins. Tannins, quercetin and catechin equivalents were used for these parameters. The antioxidant activities of the leaves and stem extracts of Buddleja saligna were determined by ABTS, DPPH, and ferrous reducing antioxidant property (FRAP) methods. Laboratory isolates of 10 bacteria species which included five Gram-positive and five Gram-negative strains were used to assay for antibacterial activity of this plant.
Results
The antioxidant activities of the leaves as determined by the ABTS and DPPH were similar to that of the stem. The flavonoids and the flavonols contents of the leaves were higher than that of the stem but the total phenols, proanthocyanidins and FRAP activities were higher in the methanol extracts of the stem. The extracts did show activity against both Gram-positive and Gram-negative bacteria. For instance, while the methanol extract of the leaves showed good activities on all the organisms except Serratia marcescens and Pseudomonas aeruginosa at MICs of between 2.5 and 5.0 mg/ml, the extract of the stem only showed activities on Bacillus cereus, Streptococcus pyrogens and Pseudomonas aeruginosa at the same concentration.
Conclusion
The results from this study indicate that the leaves and stem extracts of Buddleja saligna possess antioxidant properties and could serve as free radical inhibitors or scavenger or, acting possibly as primary antioxidants. Although, the antibacterial properties of Buddleja saligna are not as effective as the standard drugs-Chloramphenicol and Streptomycin, they still possess some activity against bacterial strains used in this study. Buddleja saligna may therefore be a good candidate for functional foods as well as pharmaceutical plant-based products.
doi:10.1186/1472-6882-9-21
PMCID: PMC2715372  PMID: 19580647

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