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1.  Antioxidant activity of Syzygium cumini leaf gall extracts  
BioImpacts : BI  2014;4(2):101-107.
Introduction: Free radicals are implicated in several metabolic diseases and the medicinal properties of plants have been explored for their potent antioxidant activities to counteract metabolic disorders. This research highlights the chemical composition and antioxidant potential of leaf gall extracts (aqueous and methanol) of Syzygium cumini (S. cumini), which have been extensively used in traditional medications to treat various metabolic diseases.
Methods: The antioxidant activities of leaf gall extracts were examined using diphenylpicrylhydrazyl (DPPH), nitric oxide scavenging, hydroxyl scavenging and ferric reducing power (FRAP) methods.
Results: In all the methods, the methanolic extract showed higher antioxidant potential than the standard ascorbic acid. The presence of phenolics, flavonoids, phytosterols, terpenoids, and reducing sugars was identified in both the extracts. When compared, the methanol extract had the highest total phenolic and flavonoid contents at 474±2.2 mg of GAE/g d.w and 668±1.4 mg of QUE/g d.w, respectively. The significant high antioxidant activity can be positively correlated to the high content of total polyphenols/flavonoids of the methanol extract.
Conclusion: The present study confirms the folklore use of S. cumini leaves gall extracts as a natural antioxidant and justifies its ethnobotanical use. Further, the result of antioxidant properties encourages the use of S. cumini leaf gall extracts for medicinal health, functional food and nutraceuticals applications.
doi:10.5681/bi.2014.018
PMCID: PMC4097971  PMID: 25035854
Polyphenols; Plants; DPPH; Gallic acid; Metabolic diseases; Drug
2.  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
3.  Quantitative analysis of antiradical phenolic constituents from fourteen edible Myrtaceae fruits 
Food chemistry  2008;109(4):883-890.
Many species of Myrtaceae are cultivated in home gardens throughout the tropics for their edible fruit, and have been used in traditional medicine to treat several inflammatory conditions. Fruit phenolics are important dietary antioxidant and anti-inflammatory constituents. We have investigated the antiradical activity, total phenolic content (TPC), and total anthocyanin content (TAC) of 14 underutilized Myrtaceae fruits, namely Eugenia aggregata, E. brasiliensis, E. luschnathiana, E. reinwardtiana, Myrciaria cauliflora, M. dubia, M. vexator, Syzygium cumini, S. curranii, S. jambos, S. javanicum, S. malaccense, S. samarangense, and S. samarangense var. Taiwan pink. An HPLC-PDA method was developed to quantify the amounts of cyanidin 3-glucoside (1), delphinidin 3-glucoside (2), ellagic acid (3), kaempferol (4), myricetin (5), quercetin (6), quercitrin (7), and rutin (8) present in MeOH extracts of the fruit. TPC ranged from 3.57 to 101 mg/g, TAC ranged from undetectable to 12.1 mg/g, and antiradical activity, measured as DPPH˙ IC50, ranged from very active (19.4 μg/ml) to inactive (389 μg/ml).
doi:10.1016/j.foodchem.2008.01.021
PMCID: PMC3040238  PMID: 21340048
HPLC-PDA; quantitative analysis; Myrtaceae; Myrciaria; Eugenia; Syzygium; antioxidant; flavonoid; anthocyanin; Folin-Ciocalteu; DPPH; polyphenolic
4.  Comparative Study of Erythrina indica Lam. (Febaceae) Leaves Extracts for Antioxidant Activity 
The present study was designed to investigate the antioxidant activity of aqueous and methanol extracts of Erythrina indica Lam leaves by in vitro methods viz. 1, 1-Diphenyl-2-Picrylhydrazyl, nitric oxide radical scavenging activity, and inhibition of lipid peroxidation by thiobarbituric acid reactive substances (TBARS) method on isolated rat liver tissues. Quantitative analysis of antioxidative components like total amount of phenolics, flavonoids, and flavonols were estimated using the spectrophotometric method. Linear regression analysis was used to calculate the IC50 value. Results showed that the aqueous and methanol extracts exhibited significant DPPH radicals scavenging activity with an IC50 value 342.59 ± 19.59, 283.24 ± 12.28 µg/mL respectively. Nitric oxide radicals were significantly scavenged by the aqueous and methanol extracts (IC50 = 250.12 ± 10.66; 328.29 ± 3.74 µg/mL). Lipid peroxidation induced by the Fe2+ was inhibited by the aqueous extract with low IC50 value (97.29 ± 2.05 µg/mL) as compared to methanol extract (IC50 = 283.74 ± 5.70 µg/mL). Both the extracts were exhibited similar quantities of total phenolics. Total flavonoids were found to be in higher quantities than total flavonols in aqueous extract as compared to methanol extract. From the results, it is concluded that the aqueous and methanol extracts of E. indica leaves possesses significant antioxidant activity that may be due to the presence of flavonoids and related polyphenolic compounds.
doi:10.4103/0975-1483.62216
PMCID: PMC3035888  PMID: 21331194
Antioxidant; Erythrina indica; gallic acid; polyphenols; radicals; rutin
5.  Antioxidant and Antibacterial Activities of Crude Extracts and Essential Oils of Syzygium cumini Leaves 
PLoS ONE  2013;8(4):e60269.
This research highlights the chemical composition, antioxidant and antibacterial activities of essential oils and various crude extracts (using methanol and methylene chloride) from Syzygium cumini leaves. Essential oils were analyzed by gas chromatography-mass spectrometry (GC-MS).The abundant constituents of the oils were: α-pinene (32.32%), β-pinene (12.44%), trans-caryophyllene (11.19%), 1, 3, 6-octatriene (8.41%), delta-3-carene (5.55%), α-caryophyllene (4.36%), and α-limonene (3.42%).The antioxidant activities of all extracts were examined using two complementary methods, namely diphenylpicrylhydrazyl (DPPH) and ferric reducing power (FRAP). In both methods, the methanol extract exhibited a higher activity than methylene chloride and essential oil extracts. A higher content of both total phenolics and flavonoids were found in the methanolic extract compared with other extracts. Furthermore, the methanol extract had higher antibacterial activity compared to methylene chloride and the essential oil extracts. Due to their antioxidant and antibacterial properties, the leaf extracts from S. cumini may be used as natural preservative ingredients in food and/or pharmaceutical industries.
doi:10.1371/journal.pone.0060269
PMCID: PMC3625181  PMID: 23593183
6.  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
7.  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
8.  Determination of Antibacterial and Antioxidant Potential of Some Medicinal Plants from Saurashtra Region, India 
Many plants used in Saurashtra folk medicine have been reported to exhibit high antibacterial and antioxidant activities. In the present study, some parts of five plants, Guazuma ulmifolia L., Manilkara zapota L., Melia azedarach L., Syzygium cumini L. and Wrightia tomentosa R.& S., were evaluated for their antibacterial activity, total phenol content, flavonoid content, 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity and phytochemical analysis, using successive extraction by cold percolation method with petroleum ether, ethyl acetate, methanol and water. In vitro antibacterial activity was evaluated against five bacterial strains viz. Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhimurium and Enterobacter aerogenes by agar well diffusion method. Among the plants screened, W. tomentosa leaf and fruit showed the best antibacterial activity. The Gram-positive bacteria were more susceptible than Gram-negative bacteria. Methanol extract of M. zapota showed the best 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity. Highest total phenol content was shown by M. zapota and S. cumini in methanol extract, while highest flavonoid content was shown by W. tomentosa stem in petroleum ether extract and ethyl acetate extract. In all the plants, cardiac glycosides and triterpenes were more as compared to other phytoconstituents.
doi:10.4103/0250-474X.57289
PMCID: PMC2865812  PMID: 20502546
Antibacterial activity; antioxidant activity; Guazuma ulmifolia L.; Manilkara zapota L.; Melia azedarach L.
9.  Attenuation of oxidative stress in U937 cells by polyphenolic-rich bark fractions of Burkea africana and Syzygium cordatum 
Background
Oxidative stress has been implicated in the progression of various diseases, which may result in the depletion of endogenous antioxidants. Exogenous supplementation with antioxidants could result in increased protection against oxidative stress. As concerns have been raised regarding synthetic antioxidant usage, the identification of alternative treatments is justified. The aim of the present study was to determine the antioxidant efficacy of Burkea africana and Syzygium cordatum bark extracts in an in vitro oxidative stress model.
Methods
Cytotoxicity of crude aqueous and methanolic extracts, as well as polyphenolic-rich fractions, was determined in C2C12 myoblasts, 3T3-L1 pre-adipocytes, normal human dermal fibroblasts and U937 macrophage-like cells using the neutral red uptake assay. Polyphenolic content was determined using the Folin-Ciocalteau and aluminium trichloride assays, and antioxidant activity using the Trolox Equivalence Antioxidant Capacity and DPPH assays. The extracts efficacy against oxidative stress in AAPH-exposed U937 cells was assessed with regards to reactive oxygen species generation, cytotoxicity, apoptosis, lipid peroxidation and reduced glutathione depletion.
Results
B. africana and S. cordatum showed enrichment of polyphenols from the aqueous extract, to methanolic extract, to polyphenolic-rich fractions. Antioxidant activity followed the same trend, which correlated well with the increased concentration of polyphenols, and was between two- to three-fold stronger than the Trolox antioxidant control. Both plants had superior activity compared to ascorbic acid in the DPPH assay. Polyphenolic-rich fractions were most toxic to the 3T3-L1 (IC50’s between 13 and 21 μg/ml) and C2C12 (IC50’s approximately 25 μg/ml) cell lines, but were not cytotoxic in the U937 and normal human dermal fibroblasts cultures. Free radical-induced generation of reactive oxygen species (up to 80%), cytotoxicity (up to 20%), lipid peroxidation (up to 200%) and apoptosis (up to 60%) was successfully reduced by crude extracts of B. africana and the polyphenolic-rich fractions of both plants. The crude extracts of S. cordatum were not as effective in reducing cytotoxic parameters.
Conclusion
Although oxidative stress was attenuated in U937 cells, cytotoxicity was observed in the 3T3-L1 and C2C12 cell lines. Further isolation and purification of polyphenolic-fractions could increase the potential use of these extracts as supplements by decreasing cytotoxicity and maintaining antioxidant quality.
doi:10.1186/1472-6882-13-116
PMCID: PMC3680320  PMID: 23714009
Antioxidant; Apoptosis; Burkea africana; Cytotoxicity; Free radicals; Glutathione; Lipid peroxidation; Oxidative stress; Polyphenols; Syzygium cordatum
10.  Phytochemical screening, antioxidants and antimicrobial potential of Lantana camara in different solvents 
Objective
To evaluate the antioxidant activity, hydrogen peroxide radicals scavenging activity, reducing power, the total phenolic and flavonoids contents, and antimicrobial and antifungal activities of methanol, ethanol and water extracts of leaves of Lantana camara (L. camara).
Methods
Methanol, ethanol and water extracts were evaluated against four Gram positive and Gram negative bacterial isolates (Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus subtilis) and two fungal strains (Aspergillus fumigatus and Aspergillus flavus). Methanol extract at different concentrations was tested for antioxidant potential and phytochemicals were determined by using spectrophotometric method.
Results
The total phenolic content was (40.859±0.017) mg gallic acid/g in the leaves of L. camara, while the total flavonoids was (53.112±0.199) mg/g dry weight. Methanol leaf extract of L. camara showed maximum antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa and was also effective against other bacterial strains as compared to ethanol and aqueous extracts of leaves. The methanol leaf extract of L. camara exhibited significant inhibition (71%) and (66%) against Aspergillus fumigatus and Aspergillus flavus respectively.
Conclusions
The methanol extract of the L. camara leaves is effective against selected bacterial and fungal strains. Its phytochemical contents have broad antimicrobial properties and the plant might be a novel source of antimicrobial drug.
doi:10.1016/S2222-1808(13)60104-8
PMCID: PMC4027341
MIC; Antioxidants; Phytochemicals; Antimicrobial; Lantana camara
11.  In vitro antimicrobial activity on clinical microbial strains and antioxidant properties of Artemisia parviflora 
Background
Artemisia parviflora leaf extracts were evaluated for potential antimicrobial and antioxidant properties. Antimicrobial susceptibility assay was performed against ten standard reference bacterial strains. Antioxidant activity was analyzed using the ferric thiocyanate and 2, 2-Diphenyl-1-Picrylhydrazyl (DPPH) assays. Radical scavenging activity and total phenolic content were compared. Phytochemical analyses were performed to identify the major bioactive constitution of the plant extract.
Results
Hexane, methanol and ethyl acetate extracts of A. parviflora leaves exhibited good activity against the microorganisms tested. The n-hexane extract of A. parviflora showed high inhibition of the growth of Pseudomonas aeruginosa, Escherichia coli and Shigella flexneri. Methanol extract showed strong radical scavenging and antioxidant activity, other extracts showed moderate antioxidant activity. The major derivatives present in the extracts are of terpenes, steroids, phenols, flavonoids, tannins and volatile oil.
Conclusions
The results obtained with n-hexane extract were particularly significant as it strongly inhibited the growth of P. aeruginosa, E. coli and S. flexneri. The major constituent of the n-hexane extract was identified as terpenes. Strong antioxidant activity could be observed with all the individual extracts. The antimicrobial and antioxidant property of the extracts were attributed to the secondary metabolites, terpenes and phenolic compounds present in A. parviflora and could be of considerable interest in the development of new drugs.
doi:10.1186/1476-0711-11-30
PMCID: PMC3582536  PMID: 23171441
Artemisia; Terpenoids; Antimicrobial; Antioxidant; Radical scavenging activity
12.  Evaluation of behavioural and antioxidant activity of Cytisus scoparius Link in rats exposed to chronic unpredictable mild stress 
Background
Various human diseases have oxidative stress as one of their component. Many herbs have been reported to exhibit properties that combat oxidative stress through their active constituents such as flavonoids, tannins, phenolic compounds etc. Cytisus scoparius (CS) Link, (Family: Leguminosae), also called Sarothamnus scoparius, has been shown in invitro experiments to be endowed with anti-diabetic, hypnotic and sedative and antioxidant activity. Therefore this study was carried out to evaluate CS for its anxiolytic, antidepressant and anti-oxidant activity in stressed rats.
Methods
60% methanolic extract of CS was quantified for phenolic content by Folin-Ciocalteau's method. Chronic unpredictable mild stress (CMS) was employed to induce stress in rats. CS (125 and 250 mg/kg, p.o) and diazepam (DZM) (2 mg/kg, p.o) was administered during the 21 day stress exposure period. Anxiolytic and antidepressant activities of CS were assessed in open field exploratory and behavioural despair paradigms, respectively. Plasma glucose and total lipids; endogenous antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT); non-enzymic-ascorbic acid and thiobarbituric acid reactive substances (TBARS) levels were measured in brain, kidneys and adrenals using standard protocols to assess the effect of CS.
Results
Total phenolic content of CS was found to be 8.54 ± 0.16% w/w. CMS produced anxiogenic and depressive behaviour in experimental rats with metabolic disturbance. Significant decrease in SOD, CAT levels and increase in lipid peroxidation level was observed in stressed rats. CS administration for 21 days during stress exposure significantly increased the ambulatory behaviour and decreased the freezing time in open field behaviour. In behavioural despair test no significant alteration in the immobility period was observed. CS also improved SOD, CAT, and ascorbic acid level and controlled the lipid peroxidation in different tissues.
Conclusion
CS possesses anti-stress and moderate anxiolytic activity which may be due, in part, to its antioxidant effect that might warrant further studies.
doi:10.1186/1472-6882-8-15
PMCID: PMC2386776  PMID: 18435835
13.  Phytochemical Analysis and Antioxidant Property of Leaf Extracts of Vitex doniana and Mucuna pruriens 
Oxidative stress and impaired antioxidant system have been implicated in the pathophysiology of diverse disease states. The phytochemical screening and antioxidant property of fresh leaves of Vitex doniana and Mucuna pruriens, used in the management and treatment of various diseases, were studied. The extracts (ethanol and distilled water) were screened for the presence of phytochemicals, and their inhibition of 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical was used to evaluate their free radical scavenging activity. Liver levels of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) in carbon tetrachloride- (CCl4) treated albino rats were also used to assess the antioxidant activity of the extracts. The animals were treated with 250 mg/kg body weight of the extracts for six consecutive days before a single dose (2.5 mL/kg body weight) of CCl4. Vitamin C was used as the standard antioxidant. Phytochemical screening revealed the presence of saponins, tannins, anthraquinones, terpenoids, and flavonoids in all the extracts, while alkaloids were detected in extracts of Vitex doniana only, and cardiac glycosides occurred in extracts of Mucuna pruriens only. All the extracts inhibited DPPH radical in a concentration-dependent manner, water extract of Vitex doniana producing highest inhibition which was not significantly different (P > .05) from vitamin C. The extracts produced a significant decrease (P < .05) in liver MDA, while the levels of SOD and CAT significantly increased (P < .05) relative to the positive control. These results are an indication of antioxidant potential of the extracts and may be responsible for some of the therapeutic uses of these plants.
doi:10.1155/2011/459839
PMCID: PMC3085303  PMID: 21547085
14.  Phytochemical Analysis and Free Radical Scavenging Activity of Medicinal Plants Gnidia glauca and Dioscorea bulbifera 
PLoS ONE  2013;8(12):e82529.
Gnidia glauca and Dioscorea bulbifera are traditional medicinal plants that can be considered as sources of natural antioxidants. Herein we report the phytochemical analysis and free radical scavenging activity of their sequential extracts. Phenolic and flavonoid content were determined. Scavenging activity was checked against pulse radiolysis generated ABTS•+ and OH radical, in addition to DPPH, superoxide and hydroxyl radicals by biochemical methods followed by principal component analysis. G. glauca leaf extracts were rich in phenolic and flavonoid content. Ethyl acetate extract of D. bulbifera bulbs and methanol extract of G. glauca stem exhibited excellent scavenging of pulse radiolysis generated ABTS•+ radical with a second order rate constant of 2.33×106 and 1.72×106, respectively. Similarly, methanol extract of G. glauca flower and ethyl acetate extract of D. bulbifera bulb with second order rate constants of 4.48×106 and 4.46×106 were found to be potent scavengers of pulse radiolysis generated OH radical. G. glauca leaf and stem showed excellent reducing activity and free radical scavenging activity. HPTLC fingerprinting, carried out in mobile phase, chloroform: toluene: ethanol (4: 4: 1, v/v) showed presence of florescent compound at 366 nm as well as UV active compound at 254 nm. GC-TOF-MS analysis revealed the predominance of diphenyl sulfone as major compound in G. glauca. Significant levels of n-hexadecanoic acid and octadecanoic acid were also present. Diosgenin (C27H42O3) and diosgenin (3á,25R) acetate were present as major phytoconstituents in the extracts of D. bulbifera. G. glauca and D. bulbifera contain significant amounts of phytochemicals with antioxidative properties that can be exploited as a potential source for herbal remedy for oxidative stress induced diseases. These results rationalize further investigation in the potential discovery of new natural bioactive principles from these two important medicinal plants.
doi:10.1371/journal.pone.0082529
PMCID: PMC3867356  PMID: 24367520
15.  Study on Phytochemical Composition, Antibacterial and Antioxidant Properties of Different Parts of Alstonia scholaris Linn. 
Advanced Pharmaceutical Bulletin  2013;3(2):379-384.
Purpose: To evaluate phytochemical composition, antibacterial and antioxidant properties of methanolic extracts of different parts viz., leaves, follicles and latex of Indian devil tree (Alstonia scholaris Linn.) R. Br. Methods: Antibacterial activities of the methanol extracts against Gram +ve (Bacillus subtilis and Staphylococcus aureus) and Gram -ve (Escherichia coli, Pseudomonas aeruginosa) bacteria were determined by well diffusion techniques. Aantioxidant profiles of methanol extracts were determined by 1,1-diphenyl-2-picryl-hydrazil (DPPH) free radical scavenging, superoxide anion radial scavenging and ferric thiocyanate reducing assays. Results: Phytochemical composition revealed abundance of flavonoids (97.3 mg QE/g DW), proanthocynidins (99.3 mg CE/g DW) and phenolics (49.7 mgGAE/g DW) in the leaf extract. Extracts of follicles and latex had comparatively very content of phenolics, flavonoids and proanthocyanidins. However, in follicle extract level of proanthocyanidins was significantly higher (46.8 mg CE/gDW). Latex extract among others exhibited most potent antibacterial activity. All the extracts displayed strong DPPH free radical and superoxide anion scavenging activities, only leaf extract displayed powerful reducing and ferrous ion chelating activities. Conclusion: Study revealed significant antioxidant activities of A. scholaris leaf, follicles and latex extracts and potential antibacterial activity of latex extract.
doi:10.5681/apb.2013.061
PMCID: PMC3848240  PMID: 24312864
Alstonia scholaris; Antibacterial; Antioxidant; Follicles; Latex; Phenolics
16.  Preliminary phytochemical screening and In vitro antioxidant activities of the aqueous extract of Helichrysum longifolium DC 
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
17.  Phytochemical studies and antioxidant activity of two South African medicinal plants traditionally used for the management of opportunistic fungal infections in HIV/AIDS patients 
Background
It has been observed that perturbations in the antioxidant defense systems, and consequently redox imbalance, are present in many tissues of HIV-infected patients. Hence, the exogenous supply of antioxidants, as natural compounds that scavenge free radicals, might represent an important additional strategy for the treatment of HIV infection. The aim of this study was therefore to analyse the phytochemical constituents and antioxidant potential of Gasteria bicolor Haw and Pittosporum viridiflorum Sims., two South African plants traditionally used for the management of opportunistic fungal infections (OFIs) in AIDS patients.
Methods
The in vitro antioxidant properties of the two plants were screened through DPPH (1,1-diphenyl-2-picrylhydrazyl), NO (nitric oxide), H2O2 (hydrogen peroxide) radical scavenging effects and reducing power assays. Phytochemical studies were done by spectrophotometric techniques.
Results
There were no significant differences in the flavonoid and proanthocyanidins contents between the leaves and bark extracts of Gasteria bicolor and Pittosporum viridiflorum respectively, while the total phenolic content of the bark extract of P. viridiflorum was significantly higher than that of G. bicolor leaf. The acetone extracts of both plants indicated strong antioxidant activities.
Conclusion
The results from this study indicate that the leaves and stem extracts of Gasteria bicolor and Pittosporum viridiflorum respectively possess antioxidant properties and could serve as free radical inhibitors, acting possibly as primary antioxidants. Since reactive oxygen species are thought to be associated with the pathogenesis of AIDS, and HIV-infected individuals often have impaired antioxidant defenses, the inhibitory effect of the extracts on free radicals may partially justify the traditional use of these plants in the management of OFIs in HIV patients in South Africa.
doi:10.1186/1472-6882-12-43
PMCID: PMC3364152  PMID: 22502778
Antioxidant; phytochemical; P. viridiflorum; G. bicolor; Opportunistic fungi; HIV/AIDS
18.  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
19.  Phytochemical, Anti-oxidant and Anthelmintic activities of various leaf extracts of Flacourtia sepiaria Roxb 
Objective
The present study was carried out to investigate the phytochemical constituents, in vitro antioxidant potential and anthelmintic activities of Flacourtia sepiaria Roxb leaves.
Methods
The dried powdered leaves of Flacourtia sepiaria were extracted using petroleum ether, chloroform, ethyl acetate and methanol by a soxhlet extractor and preliminary phytochemical screening was performed using standard protocols. All the extract was evaluated for their potential antioxidant activities using test such as DPPH, superoxide anion radical, hydroxyl radical, nitric oxide radical scavenging abilities, ferrous chelating ability and total phenolic and flavanoid content. Anthelmintic activity of extract was screened in adult Indian earthworm model.
Results
Preliminary screening revealed the presence of bioactive compounds especially phenolics, tannins and terpenoids in all extracts. The phenolic and flavanoid content was highest in methanolic extract and lowest in petroleum ether extract. The paralytic (9.46±0.212) and death time (31.43±0.148) of methanolic extract was found to be significant (P<0.05) when compared with paralytic (7.33±0.206) and death time (18.60±0.229) of standard piperazine citrate at 100 mg/mL concentration.
Conclusions
The results of the present study indicate that the leaf extracts of Flacourtia sepiaria exhibited strong antioxidant activity and possess significant anthelmintic activity and thus it is a good source of antioxidant and anthelmintic constituents.
doi:10.1016/S2221-1691(13)60184-7
PMCID: PMC3805094  PMID: 24093785
Antioxidant; Anthelmintic; Flacourtia sepiaria; Piperazine citrate.
20.  Estimation of total phenol and in vitro antioxidant activity of Albizia procera leaves 
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
21.  Phytochemical screening, antimicrobial and antioxidant efficacy of different extracts of Rumex dentatus L. - A locally used medicinal herb of Kashmir Himalaya 
Objective
To elucidate the antimicrobial and antioxidant activities of Rumex dentatus L. (R. dentatus) along with its phytochemical analysis.
Methods
Agar disk diffusion method for antimicrobial activity and DPPH, riboflavin photo-oxidation, deoxyribose and lipid peroxidation assay for antioxidant activity.
Results
The antimicrobial and antioxidant activities of different concentrations of five R. dentatus extracts were tested against different clinical bacterial strains (Shigella flexneri, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhimurium) and fungal strains (Aspergillus versicolor, Aspergillus flavus, Accremonium spp., Penicillium dimorphosporum, Candida albicans, Candida parapsilosis, and Candida kruesie). Among all extracts, the butanol extract showed strong antibacterial activity against Klebsiella pneumoniae (inhibition zone diameter of 20 mm) and aqueous extract showed no activity against any of the bacterial strains. While as in case of the fungal strains, the maximum antifungal activity was observed against Aspergillus flavus by aqueous extract. The antioxidant activity revealed that the extracts exhibited scavenging effect in concentration-dependent manner on superoxide anion radicals and hydroxyl radicals. The phytochemical tests carried out with the crude extracts of R. dentatus showed the presence of flavonoids, terpenoids, alkaloids, saponins, tannins, anthraquinones and cardiac glycosides in it. The total phenolic content of these extracts was estimated quantitatively from standard calibration curve of gallic acid and it varied from 145 µg/mg in butanol extract to 45 µg/mg in petroleum ether extract.
Conclusions
It can be concluded that the plant has got a broad spectrum antimicrobial and antioxidant activity and could be used as a potential alternative for treating various diseases.
doi:10.1016/S2222-1808(13)60097-3
PMCID: PMC4027333
Antimicrobial activity; Antioxidant activity; Rumex dentatus; Medicinal herb
22.  Cytotoxic chalcones and antioxidants from the fruits of a Syzygium samarangense (Wax Jambu) 
Food chemistry  2008;107(2):813-819.
Bioassay-guided fractionation of the methanolic extracts of the pulp and seeds of the fruits of Syzygium samarangense Merr. & Perry (Blume) led to the identification of four cytotoxic compounds and eight antioxidants on the basis of HPLC-PDA analysis, MS, and various NMR spectroscopic techniques. Three C-methylated chalcones, 2′,4′-dihydroxy-3′,5′-dimethyl-6′-methoxychalcone (1), 2′,4′-dihydroxy-3′-methyl-6′-methoxychalcone (stercurensin, 2), and 2′,4′-dihydroxy-6′-methoxychalcone (cardamonin, 3), were isolated and displayed cytotoxic activity (IC50 = 10, 35, and 35 μM, respectively) against the SW-480 human colon cancer cell line. Also a number of known antioxidants were obtained including six quercetin glycosides: reynoutrin (4), hyperin (5), myricitrin (6), quercitrin (7), quercetin (9), and guaijaverin (10), one flavanone: (S)-pinocembrin (8), and two phenolic acids: gallic acid (11) and ellagic acid (12).
doi:10.1016/j.foodchem.2007.08.086
PMCID: PMC3282680  PMID: 22359426
Syzygium samarangense; chalcones; flavonoids; cytotoxic activity; antioxidants
23.  Antioxidant Capacities, Phenolic Profile and Cytotoxic Effects of Saxicolous Lichens from Trans-Himalayan Cold Desert of Ladakh 
PLoS ONE  2014;9(6):e98696.
Fourteen saxicolous lichens from trans-Himalayan Ladakh region were identified by morpho-anatomical and chemical characteristics. The n-hexane, methanol and water extracts of the lichens were evaluated for their antioxidant capacities. The lichen extracts showing high antioxidant capacities and rich phenolic content were further investigated to determine their cytotoxic activity on human HepG2 and RKO carcinoma cell lines. The ferric reducing antioxidant power (FRAP), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH) and nitric oxide (NO) radical scavenging capacities and β-carotene-linoleic acid bleaching property exhibited analogous results where the lichen extracts showed high antioxidant action. The lichen extracts were also found to possess good amount of total proanthocyanidin, flavonoid and polyphenol. The methanolic extract of Lobothallia alphoplaca exhibited highest FRAP value. Methanolic extract of Xanthoparmelia stenophylla showed the highest ABTS radical scavenging capacity. The n-hexane extract of Rhizoplaca chrysoleuca exhibited highest DPPH radical scavenging capacity. Highest antioxidant capacity in terms of β-carotene linoleic acid bleaching property was observed in the water extract of Xanthoria elegans. Similarly, Melanelia disjuncta water extract showed highest NO scavenging capacity. Among n-hexane, methanol and water extracts of all lichens, the methanolic extract of Xanthoparmelia mexicana showed highest total proanthocyanidin, flavonoid and polyphenol content. From cytotoxic assay, it was observed that the methanolic extracts of L. alphoplaca and M. disjuncta were exhibiting high cytotoxic effects against cancer cell growth. Similarly, the water extract of Dermatocarpon vellereum, Umbilicaria vellea, X. elegans and M. disjuncta and the methanolic extract of M. disjuncta and X. stenophylla were found to possess high antioxidant capacities and were non-toxic and may be used as natural antioxidants for stress related problems. Our studies go on to prove that the unique trans-Himalayan lichens are a hitherto untapped bioresource with immense potential for discovery of new chemical entities, and this biodiversity needs to be tapped sustainably.
doi:10.1371/journal.pone.0098696
PMCID: PMC4061001  PMID: 24937759
24.  Phytochemical analysis and evaluation of leaf and root parts of the medicinal herb, Hypochaeris radicata L. for in vitro antioxidant activities 
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
Hypochaeris radicata; Phytochemical analysis; In vitro antioxidant activities
25.  Evaluation of Antioxidant Potential of “Maltese Mushroom” (Cynomorium coccineum) by Means of Multiple Chemical and Biological Assays 
Nutrients  2013;5(1):149-161.
Cynomorium coccineum is an edible, non-photosynthetic plant widespread along the coasts of the Mediterranean Sea. The medicinal properties of Maltese mushroom—one of the oldest vernacular names used to identify this species—have been kept in high regard since ancient times to the present day. We evaluated the antioxidant potential of fresh specimens of C. coccineum picked in Sardinia, Italy. Both aqueous and methanolic extracts were tested by using multiple assay systems (DPPH, FRAP, TEAC, ORAC-PYR). Total phenolics and flavonoids were also determined. Gallic acid and cyanidin 3-O-glucoside were identified as the main constituents and measured. Both extracts showed antioxidant capacities; ORAC-PYR assay gave the highest antioxidant value in both cases. The methanolic extract was further investigated with in vitro biological models of lipid oxidation; it showed a significant activity in preventing cholesterol degradation and exerted protection against Cu2+-mediated degradation of the liposomal unsaturated fatty acids. Results of the present study demonstrate that the extracts of C. coccineum show a significant total antioxidant power and also exert an in vitro protective effect in different bio-assays of oxidative stress. Therefore, Maltese mushroom can be considered a valuable source of antioxidants and phytochemicals useful in the preparation of nutraceuticals and functional foods.
doi:10.3390/nu5010149
PMCID: PMC3571642  PMID: 23344249
plant-based foods; antioxidant; nutraceuticals; Cynomorium coccineum; fungus melitensis; Maltese mushroom

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