Superoxide Radical Scavenging Activity Research Articles

Ultrasound pretreatment of fermented milk containing probiotic Lactobacillus plantarum AF1: Carbohydrate metabolism and antioxidant activity

AbstractThe effect of ultrasound (100 W, 30 kHz, 25% amplitude for 5, 10, and 15 min; US) on growth, carbohydrate metabolism, and antioxidant activity of probiotic Lactobacillus plantarum AF1 was investigated during milk fermentation. The antioxidant activity of L. plantarum AF1 was measured by 2,2′‐azinobis‐(3‐ethylbenzothiazoline‐6‐sulfonate) (ABTS), 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH), inhibition of the autoxidation of ascorbate, reducing ability, and superoxide anion radical scavenging activity tests. The results indicated that the cell membrane permeability of sonicated samples was increased 88–94% compared to control. Additionally, β‐galactosidase activity and population of Lactobacillus strain increased with increasing ultrasound treatment time during the fermentation after the treatment. However, no significant differences were observed in the final counts of the control and the sonicated samples at the end of fermentation (24 hr). Although the lactose content was decreased significantly (p ≤ .05) by extending sonication time, the amounts of glucose, galactose, and lactic acid enhanced significantly (p ≤ .05) as the time of fermentation and sonication proceeded. The antioxidant activity was also enhanced significantly (p ≤ .05) with increase in sonication time during 24 hr fermentation after the treatment. Therefore, the lowest and the highest antioxidant activities were seen in the control sample and the US‐15 min sonicated milk at the end of fermentation, respectively.Practical applicationsThe dairy industry is seeking methods to increase nutritional and quality of dairy products. Recently, the ultrasound has attracted significant attention in food processing generally because of the recent consumers’ trend toward functional foods. The present study provides an investigation about the efficiency of ultrasound to improve the metabolism and growth of Lactobacillus plantarum during fermentation of milk and therefore increase the quality and antioxidant activity of milk. The results show that ultrasound is a potential technique for improve metabolism of Lactobacillus strain and thus increase antioxidant activity and quality of fermented milk. Consequently, the use of ultrasound will increase the health promoting effects and quality of fermented dairy products. These results will assist in designing ultrasound procedures for optimizing the general quality of fermented milk.

In Vitro Antioxidant Activity of Hydrophila auriculata Leaves Extract and Silver Nanoparticles

Antioxidant activity of Hydrophila auriculata leaf extract and silver nanoparticles (AgNPs) were carried out for proving its utility in free radical mediated diseases. The flower extract and silver nanoparticles were screened for in vitro antioxidant activity by oxygen radical scavenging such as DPPH, total antioxidant assay and superoxide radical scavenging activity at different concentrations. The antioxidant potential was found to be dose dependent. AgNPs possess probable antioxidant activity as compared with Hydrophila auriculata leaf extract and near to the standard. It is an obvious indication that the resulting activity of the AgNPs is not due to any capping agents, which are found in minute amounts but is due to the elemental silver.

Evaluation of antioxidant activities of various solvent extract from Sargassum serratifolium and its major antioxidant components.

Sargassum serratifolium has been known to contain a high level of meroterpenoids as antioxidant components. We investigated antioxidant activities and active components in various solvent extracts from S. serratifolium. Ethyl acetate, ethanol, and methanol extracts showed relatively strong DPPH, ABTs, and superoxide radical scavenging activities. Hexane and ethyl acetate extract showed the strongest hydroxyl radical and reactive oxygen species (ROS), respectively, scavenging activities. Sargahydroquinoic acid (SHQA), sargachromanol (SCM) and sargaquinoic acid (SQA) were main antioxidant components in S. serratifolium. Ethanol extract showed the highest levels of SHQA, SCM, and SQA which comprised to be 227 ± 6.31 mg/g. SHQA and SCM exhibited stronger antioxidant capacities than SQA based on lower IC50 values in ROS, DPPH, ABTs, and superoxide radical scavenging assays. The result showed that ethanol is the most efficient extracting solvent for the active components from S. serratifolium and the plant has the potential as a natural antioxidant.

Utilization of Saccharina japonica as a Solid-Fermented Substrate for the Production of Bioactive Ethanolic Extract

Marine biomass, Saccharina japonica was fermented as a solid-fermented substrate by Monascus spp. for the production of bioactive ethanolic extract. The 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-Azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS), superoxide anion radical scavenging and DNA protection activities of ethanolic extracts obtained from S. japonica fermented by M. purpureus (SjMp) showed the highest activity followed by M. kaoliang fermented S. japonica (SjMk) and unfermented (SjU) extract at 10 mg/mL concentration. Kinetic analysis revealed that ethanolic extract of fermented S. japonica inhibited the α-amylase competitively but α-glucosidase displayed competitive inhibition at 10 mg/mL concentration and non-competitive inhibition at 1 mg/mL concentration. The Linewaver-Burk kinetics analysis revealed that ethanolic extract of SjMk showed significantly higher Km value (4.55 mg) than SjMP (2.74 mg) followed by SjU (0.35 mg) at 10 mg/mL concentrations in α-amylase inhibition. But incase of α-glucosidase inhibition, the Km value was maximum in ethanolic extract of SjMp (2.30 mg) with Vmax (0.29 mg/min) at 10 mg/mL concentrations. Both of the fermented and unfermented samples did not show toxic effect on Caco-2 cells. The amino acid compositional analysis showed that fermentation process changed free amino acids contents in ethanolic extract. Monascus-fermented ethanolic extract showed antibacterial activity on Aeromonas hydrophila and Staphylococcus aureus. So, Monascus spp. fermented S. japonica extract played an important role in prevention of free radicals formation and hyperglycemia which can be applied in biofunctional food formulation.

EVALUATION OF IN VITRO ANTIOXIDANT ACTIVITY OF A MEDICINAL HERB, WEDELIA CHINENSIS (OSBECK) MERRILL.

Objective: The objective is to evaluate the antioxidant activity of the methanolic leaf extract of Wedelia chinensis.Methods: In vitro antioxidant activity was evaluated by studying 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, superoxide radical scavenging activity, ferric reducing ability of plasma (FRAP) scavenging activity, hydroxyl radical scavenging activity, metal chelating activity, and total antioxidant activity. Ascorbic acid, BHT, quercetin, and EDTA were used as standards for the experiments.Results: Inhibition concentration50 values observed for DPPH radical scavenging, superoxide radical scavenging, and hydroxyl radical scavenging activities were determined to be 62.5, 769.23, and 617.28 μg/ml, respectively and that of FRAP scavenging, metal chelating, and total antioxidant activities were found to be 343 μmol Fe (II)/g, 79.56 mg EDTA/g extract, and 398.17 mg ascorbic acid eq/g extract, respectively.Conclusion: The results clearly indicate that methanolic leaf extract of the study species W. chinensis is effective in scavenging free radicals and has the potential to be a powerful antioxidant.

EVALUATION OF THE ANTIOXIDANT ACTIVITY OF FRUIT EXTRACTS OF INDIGENOUS MEDICINAL PLANT, ZIZYPHUS XYLOPYRUS (RETZ.) WILLD

Aim: This study aims to investigate the antioxidant activity of fruit extracts of an indigenous medicinal plant, Zizyphus xylopyrus (Retz.) Willd. Materials and Methods: The coarsely powdered fruit of Z. xylopyrus Willd. was successfully extracted first with petroleum ether and chloroform, followed by ethyl acetate and at last ethanol. The total flavonoids content (TFC), total phenolic content (TPC), and total tannin content (TTC) were estimated using quercetin and tannic acid equivalents as standard. The antioxidant activity of different extracts was determined by different standard in vitro methods including. Results: Among all extracts, ethyl acetate extract (ZXEAE) possessed potent antioxidant activity, namely hydroxyl radical (OH°) scavenging assay (IC50 128.1 ± 0.63 μg/mL), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (DPPH°) scavenging assay (IC50 101.2 ± 1.75 μg/mL), nitric oxide radical (NOo) scavenging assay (IC50 122.5 ± 2.04 μg/mL), superoxide radical (O2¯) scavenging activity assay (IC50 64.1 ± 1.24 μg/mL), and inhibition of lipid peroxidation (98.6 ± 2.05 μg/mL). Study results suggested that antioxidant activity of ZXEAE might be due to presence of polyphenols, namely TFC (31.23 ± 0.39 quercetin equivalent [QE] mg/g extract), TPC (198.14 ± 0.69 gallic acid equivalent [GAE] mg/g extract), and TTC (18.38 ± 1.79 GAE mg/g extract). Conclusion: The present study demonstrated the presence of flavonoids in Z. xylopyrus fruits showing marked antioxidant activity, which can be used for establishing the hepatoprotective potential of Z. xylopyrus

Effects of in vitro saliva, gastric and intestinal digestion on the chemical properties, antioxidant activity of polysaccharide from Artocarpus heterophyllus Lam. (Jackfruit) Pulp

Polysaccharides are one of the most widely distributed and abundant compounds in plants, animals and microorganisms. In this study, we investigated the effects of in vitro saliva, gastric and intestinal digestion on the reducing sugar contents, molecular weight and structure change of polysaccharide from Artocarpus heterophyllus Lam. Pulp (JFP-Ps). Digested samples of JFP-Ps were also subject to antioxidant assays including DPPH scavenging assay, hydroxyl radical scavenging assay, superoxide radical scavenging assay and ferric reducing power assay. Results revealed that in vitro digestion could significantly increase the reducing sugar contents and decrease the molecular weight of JFP-Ps. FT-IR results showed that structure and conformation of JFP-Ps were remarkably altered after simulated digestion. The intestinal digested samples of JFP-Ps exhibited obvious DPPH, hydroxyl radical, and superoxide radical scavenging activities in a time-dependent manner during digestion, but with a relatively low reducing power. These results indicated that digestion remarkably affected the chemical properties, structure and antioxidant activity of JFP-Ps during gastrointestinal digestion, and might help maximize the potential applications of JFP-Ps as a dietary source of antioxidant phytochemicals that survive the gastrointestinal digestion process.

Optimization of enzyme assisted extraction of polysaccharides from pomegranate peel by response surface methodology and their anti-oxidant potential

ObjectiveTo optimize the enzymatic extraction conditions of polysaccharides from pomegranate peel using response surface methodology (RSM), and invest the anti-oxidant activity of pomegranate peel polysaccharide in vitro for seeking novel biological components used as pharmaceutical products and functional foods. MethodsEffects of enzymolysis time, ratio of water to raw material, and the dosage of cellulase on the extraction yield of pomegranate peel polysaccharide were fully examined by Box-Behnken design of RSM. DPPH radical scavenging activity, hydroxyl radicals scavenging activity, superoxide anion radical scavenging activity, and reducing power assay were measured by microplate reader. ResultsThe optimal extraction conditions were as follows: enzymolysis temperature 55 °C, pH 5.0, enzymolysis time 88 min, ratio of water to raw material 22:1 mL/g, and dosage of cellulose 0.93%. Under the optimal extraction conditions, the yield of pomegranate peel polysaccharide was (22.31 ± 0.07)%, which was well matched with the predicted value 22.35% of the Box-Behnken design model. Also, pomegranate peel polysaccharide demonstrated appreciable anti-oxidant potential on DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging and reducing power in vitro. ConclusionsAn optimized enzymolysis assisted method was proposed for the extraction of pomegranate peel polysaccharide, which can be used as a good anti-oxidant.

Structural Features of Sulfated Glucuronomannan Oligosaccharides and Their Antioxidant Activity.

Glucuronomannan oligosaccharides (Gs) were derived from fucoidan, which was extracted from the brown alga Sargassum thunbergii. Sulfated glucuronomannan oligosaccharides (SGs) were obtained by the sulfation of Gs. NMR techniques were used to reveal that the order of sulfation was Man-C6 > Man-C4 > Man-C1R > GlcA-C3 > Man-C3 > GlcA-C2. Finally, the antioxidant activities (hydroxyl radical scavenging activity, superoxide radical scavenging activity, reducing power and DPPH radical scavenging activity) of Gs and SGs were determined. The findings showed that the higher the degree of polymerization, the better the activity, except for the hydroxyl radical scavenging activity. In addition, the higher the sulfate content, the lower the activities for the reducing power and DPPH radical scavenging activity. Opposite results were found for the superoxide radical scavenging activity. Finally, compared with fucoidan, most Gs and SGs had higher antioxidant activity, suggesting that they might be good candidates for antioxidants.

Application of multivariate statistical techniques to assess the phenolic compounds and the in vitro antioxidant activity of commercial grape cultivars

AbstractMultivariate statistical techniques were used to differentiate two popularly consumed grape cultivars in Taiwan, Kyoho (Vitis labruscana) and American wild bunch (Vitis aestivalis), based on their phenolic content and antioxidant properties. Total phenol and flavonoid contents, and polyphenolic profile were estimated by the Folin‐Ciocalteu, colorimetric method, and ultra performance liquid chromatography, respectively. Antioxidant activities were determined by 2,2 diphenyl‐1‐picrylhydrazyl, 2,2′‐azinobis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) scavenging activity, ferric reducing antioxidant power (FRAP), nitric oxide scavenging activity (NOSA), hydrogen peroxide scavenging activity, and total antioxidant activity methods. Data were processed using multivariate analysis of variance, discriminant analysis (DA), principal component analysis (PCA), and cluster analysis. Variations in polyphenolic compositions were attributed to higher TFC (3.42 mg QE/g), ABTS• (79.08%), NOSA (27.62%), total antioxidant (77.91%), and superoxide (46.55%) radical scavenging activities of Kyoho grape extracts; and higher inhibition capacity by DPPH• (97.26%), FRAP (4.66 mM FeSO4/g), and hydrogen peroxide (40.52%) scavenging methods for American wild bunch grape extracts. Two discriminant functions in DA reported the contribution of each phenolic and antioxidant activity in discriminating the cultivars. The results of PCA analysis reported 91.35% of the total variance, which was a suitable approach for evaluating similarities between both cultivars. Statistical analyses confirmed Kyoho and American wild bunch grape extracts differed from each other in terms of polyphenolic composition and antioxidant activities. Therefore, the application of multivariate statistical techniques used in the study provided further understanding on assessment of polyphenols and antioxidant activities of these grape cultivars.

IN VITRO ANTIOXIDANT POTENTIAL AND HEPATOPROTECTIVE ACTIVITY OF TAXUS WALLICHIANA

Objectives: Taxus wallichiana, the member of family Taxaceae is used for the cure of various ailments by a large ethnic population native to the forests in Kashmir valley. The aim of this study is to evaluate antioxidant potential and hepatoprotective action of T. wallichiana as claimed by the ethnic population segment. Methods: Antioxidant potential of different T. wallichiana extracts was evaluated by estimating the total phenolics, 1,1-diphenyl -2-picrylhydrazyl (DPPH), reducing power, microsomal lipid peroxidation (LPO), and hydroxyl radical scavenging activity through standard protocols. Hepatoprotective activity of T. wallichiana was determined by evaluation of liver marker enzyme (aspartate transaminase, alanine transaminase, and alkaline phosphatase) levels using standard protocols and by histological analysis of rat liver tissues of different animal groups against CCl4 induced liver damage. Results: Phenolic content expressed as gallic acid equivalents (GAE/g) was found maximum in case of ethanolic extract (153.34 mg GAE/g), followed by extracts of methanol (149.24 mg GAE/g), hexane (134 mg GAE/g), ethyl acetate (128.9 mg GAE/g), chloroform (121.37 mg GAE/g), and aqueous (113.00 mg GAE/g). At concentration of 700 μg/ml, DPPH radical scavenging activity of methanol extract was 88.29% with IC50 (212.00 μg/ml), aqueous (82.47%) with IC50 (258.29 μg/ml), and ethyl acetate (79.57%) with IC50 (301.80%μg/ml). The reducing power of the extracts increased in a concentration dependent manner. At concentration of 700 μg/ml, 79%, 68%, and 56% inhibition was observed with extracts of methanol, aqueous, and ethyl acetate on microsomal LPO with IC50 values of 126.09, 168.83, and151.96 μg/ml, respectively. Superoxide radical scavenging activity of T. wallichiana extracts increased in a dose-dependent manner with IC50 values 170.30 μg/ml (methanol), 257.00 μg/ml (aqueous), and 297.55 μg/ml ethyl acetate extract. T. wallichiana extracts exhibited antioxidant effects on calf thymus DNA damage. The results further depict the hepatoprotective action of T. wallichiana extract at doses of 100 and 300 mg/kg and were comparable with that of standard treatment comprising 50 mg/kg-day of Vitamin C, a known hepatoprotective agent. The data were substantiated with histopathological studies of rat liver sections. Conclusion: These results comprehensively depict that T. wallichiana extracts are endowed with the free radical sequestering potential and be employed as potential antioxidant and hepatoprotective arsenal against many oxidative stress linked diseases.

Anti-ulcerogenic activity of dentatin from clausena excavata Burm.f. against ethanol-induced gastric ulcer in rats: Possible role of mucus and anti-oxidant effect

BackgroundClausena excavata Burm.f. (Rutaceae) has been used for the treatment of stomach disorders including peptic ulcer. PurposeIn this study, we aimed to investigate dentatin isolated from C. excavata Burm.f., for anti-ulcer activity against ethanol ulcer model in rats. MethodsGastric acid output, ulcer index, serum profile, histological evaluation using Hematoxylin and eosin (HE), periodic acid Schiff base stainings and immunohistochemical localization for heat shock proteins 70 (HSP70) were all investigated. Possible involvement of reduced glutathione (GSH), lipid peroxidation, prostaglandin E2 (PGE2), superoxide dismutase (SOD) enzymes, radical scavenging, and anti-Helicobacter pylori activity were investigated. ResultsDentatin showed anti-secretory activity against the pylorus ligature model and protected the gastric mucosa from ethanol ulceration, as revealed by the improved macroscopic and histological appearance. Dentatin significantly increased the gastric homogenate content of PGE2 GSH and SOD. Dentatin inhibited the lipid peroxidation as revealed by the reduced gastric content of malondialdehyde (MDA). Moreover, dentatin up-regulated HSP70 expression. However, dentatin showed insignificant anti-H. pylori activity. ConclusionDentatin possesses gastro-protective activity, which could be attributed to the anti-secretory, mucus production, anti-oxidant, and HSP70 activities.

PHYTOCHEMICAL SCREENING AND EVALUATION OF IN VITRO ANTIOXIDANT POTENTIAL OF IMMATURE PALMYRA PALM (BORASSUS FLABELLIFER LINN.) FRUITS

Objective: The present study was aimed to evaluate the in vitro antioxidant properties of immature Palmyra palm fruits which have been traditionally used for the treatment of diabetes. The qualitative phytochemical screening and quantitative estimation of total phenolic and flavonoids contents in the ethanolic extract were performed to substantiate the antioxidant and medicinal claims.Methods: Immature palmyra palm fruits were collected, authenticated, dried and powdered in an electrical grinder. The powdered fruits were delipidated in petroleum ether and soxhilation using ethanol to extract the active ingredients. Qualitative phytochemical screening, total phenolic and flavonoid contents were carried out by established methods. The in vitro antioxidant potentials were performed by diphenyl-2-picrylhydrazyl, azino-bis(3-ethylbenzothiazoline-6-sulphonic acid, nitric oxide and superoxide anion scavenging assays.Results: The phytochemical screening showed the presence of alkaloids, flavonoids, glycosides, saponins, tannins, phytosterols, triterpenoids and phenols in the immature palmyra palm fruits extract. The total phenolic and flavonoid contents in the fruits extract was found to be 104.00±0.02 μg gallic acid equivalents/100 mg of fruits extract and 98.45±0.03 μg quercetin equivalents/100 mg, respectively. The percentage inhibition of DPPH radicals range from 35 to 70% at a concentration ranges from 200-1000μg/ml. Similarly, the percentage of inhibition of ABTS radicals was found to be in the range of 40 to 75.5%. The nitric oxide scavenging activity of the fruits extract ranges from 45 to 76% whereas the superoxide radical scavenging activity ranges from 43 to 83%.Conclusion: The observed significant free radical scavenging activity along with increased total phenolic as well as flavonoid contents suggest that the immature fruits may be considered as a potential source for the identification of pharmacologically active phytochemicals capable of controlling oxidative stress.

Effect of vibrating-type ultrafine grinding on the physicochemical and antioxidant properties of Turkish galls in Uyghur medicine

We evaluated the influence of a vibrating-type ultrafine grinding treatment on the physical and chemical properties and antioxidant activity of Turkish gall powder (TGP) with particle sizes of >450, 400–250, 250–100, 100–50, and < 50 μm. The physical properties were analyzed and we found that the small particle size of the TGP increased the bulk (from 0.68 to 0.32 g/cm3) and tapped densities (from 0.83 to 0.70 g/cm3) and increased the repose (from 35.43° to 56.28°), slide (from 35.54° to 59.23°), and crash angles (from 35.68° to 42.91°). The total ash content (from 2.34% to 1.75%) decreased, but the moisture index and water solubility index were improved. The extract of TGP with a particle size of <50 μm indicated that highest gallic acid content (9.47 mg/g), methyl gallate content (34.78 mg/g) and ellagic acid contents (0.79 mg/g) accompanied with the best DPPH, hydroxyl radical, and superoxide radical scavenging activities (p < .05). Moreover, the FTIR analysis suggested that the ultrafine grinding did not damage the main structure of the components as the particle sizes decreased. The SEM images demonstrated the shape of the TGP with different particle sizes and surface morphology. The microscopic identification exhibited the vessels, oil cells, starch grains, and cluster crystals, which provided referential information in evaluating the Turkish gall quality.

Ultrasonic-Assisted Extraction of Natural Yellow Pigment from Physalis pubescens L. and Its Antioxidant Activities

Physalis pubescens L. is rich in natural pigments but has not yet been fully utilized. Ultrasound-assisted extraction of yellow pigment from Physalis pubescens L. was investigated by response surface methodology in this study. Optimal parameters were ultrasonic power of 29.21%, ultrasonic time of 14.41 min, and ultrasonic interval time of 10.55 s. The yield was 0.193% under optimal parameters. FRAP, ABTS, and superoxide radical scavenging activity of the yellow pigment were 6.11 ± 0.22 mmol/g, 2.80 ± 0.27 mmol/g, and 57281.5 ± 2749.5 U/g, respectively. The results showed that the yield of yellow pigment could be improved by ultrasonic-assisted extraction and the yellow pigment extracted by ultrasound had antioxidant activity.

The effect of polyethylene glycol-induced drought stress on photosynthesis, carbohydrates and cell membrane in Stevia rebaudiana grown in greenhouse

Drought stress is one of the major environmental stresses that limit crop production in arid regions. A greenhouse culture experiment was conducted to evaluate the response of an agronomically and economically important sweet medical herb (Stevia rebaudiana) to polyethylene glycol (PEG 6000)-induced drought stress (5, 10, and 15% (w/v) PEG, equivalent to leaf water potentials of − 0.49, − 1.40 and − 2.93 MPa, respectively) for 1 month. Plant mass, a major determinant of Stevia yield, showed a reduction after PEG treatments. PEG-reduced photosynthesis traits included the maximal quantum yield of photosystem II (Fv/Fm), efficiency of photosystems I and II (PIabs), intercellular CO2, net photosynthesis, chlorophylls, carotenoids and water use efficiency, followed by the reduction of carbohydrates. Under PEG treatment, the reactive oxygen species (ROS) accumulation occurred and plants exhibited an increase in H2O2 generation. Consequently, an increase in malondialdehyde and electrolyte leakage was evident in PEG treatment, indicating membrane lipid peroxidation. In PEG-treated plants, the ROS accumulation was accompanied by an increase in activity of some enzymatic and non-enzymatic antioxidants. Leaf extracts of PEG-treated plants showed lower superoxide anion, hydroxyl and nitric oxide radical scavenging activity than control plants. Drought stress also caused the accumulation of the compatible solutes proline and glycine betaine. Collectively, the results demonstrated that PEG-induced oxidative stress, due to insufficient antioxidant mechanisms, provoked damages to cell membrane and photosynthetic apparatus, with consequently reduced carbohydrates and plant growth. These results are of basic importance as vegetative growth is the major determining criterion for Stevia crops and adequate irrigation is crucial for obtaining higher yield.

Preparation of proanthocyanidin–chitosan complex and its antioxidant and antibacterial properties

Chitosan (CS) is a natural polysaccharide with beneficial effects on human health. To further strengthen its biological properties, a CS-based complex was prepared by combining it with proanthocyanidin (PA) and named it as PA–CS in this study. The successful preparation of PA–CS was confirmed by Fourier transform infrared spectroscopy. The size of PA–CS particles was distributed between 10 and 100 µm with a mean value of 30.6 µm and low pH and high salt concentration could increase their swelling ratio. The loading capacity of PA in PA–CS particles was 82.2 ± 1.7 mg/g, and an increased release of PA from the complex occurred at low pH and high salt concentration. After combination, PA–CS complex exhibited better antioxidant activity than CS, which is a result of the synergistic effects of PA and CS. The reducing power and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of PA–CS complex are mainly attributed to PA, whereas the superoxide radical scavenging activity of PA–CS complex largely depends on CS. Moreover, PA–CS complex showed similar diameters of inhibitory zone and identical minimal inhibitory concentrations compared with CS, suggesting that the antibacterial potential of PA–CS complex is similar to that of CS. In addition, after 20 days storage, PA–CS complex did not exhibit significant changes in DPPH radical scavenging activity and antibacterial activity, indicating the good stability of this complex.

Earthworm Grazed-Trichoderma harzianum Biofortified Spent Mushroom Substrates Modulate Accumulation of Natural Antioxidants and Bio-Fortification of Mineral Nutrients in Tomato.

The present investigation was aimed at evaluating the impact of earthworm grazed and Trichoderma harzianum biofortified spent mushroom substrate (SMS) on natural antioxidant and nutritional properties of tomato. Results of the investigation reveal that earthworm grazing and T. harzianum bio-fortification led to significant improvement in the physico-chemical properties of fresh SMS and its application increased the accumulation of natural antioxidants and mineral content in tomato as compared to either T. harzianum biofortified SMS or fresh SMS. In particular, the earthworm grazed, T. harzianum biofortified SMS (EGTHB-SMS) was found to inhibit lipid peroxidation and protein oxidation with significant increase in total polyphenol and flavonoid content in tomato. Further, it increased Fe2+/Fe3+ chelating activity, superoxide anion radical scavenging activity compared to other treatments. The results thus suggest an augmented elicitation of natural antioxidant properties in tomato treated with EGTHB-SMS, resulting in a higher radical scavenging activity, that is highly desirable for human health. In addition, the use of SMS to enhance the nutritional value of tomato fruits becomes an environment friendly approach in sustainable crop production.

Phytochemical analysis, antioxidant and antimicrobial activity of wild and in vitro derived plants of Ceropegia thwaitesii Hook - An endemic species from Western Ghats, India.

Ceropegia thwaitesii Hook (Asclepiadaceae), an endemic plant species, due to habitat destruction and over exploitation has a very restricted distribution in the Western Ghats of Tamil Nadu, India. The present wrok aimed to determine the chemical composition, the total phenolic (TPC), flavonoid (TFC) and tannin content (TEC), and to assess the antioxidant properties of various extracts of in vivo plants (IVP) and in vitro regenerated plants (IRP) of C. thwaitesii. Some phenolic compounds like gallic acid, cathechol, vanillin and salicylic acid were identified and quantified by HPLC. All the extracts possessed relevant radical scavenging activity on DPPH, Superoxide radical scavenging activity, and Nitric oxide radicals as well as total antioxidant ability. DPPH assay of in vitro methanol stems extracts and ethanol leaves extracts revealed the best antioxidant properties with important IC50 values of 0.248 ± 0.45 µg/mL and 0.397 ± 0.67 µg/mL, respectively, whereas in vivo chloroform stems extracts showed a lower antioxidant activity (IC50 of 10.99 ± 0.24 µg/mL). The IRP methanol extracts of stem and leaves had good inhibitory activity against all tested microorganisms in a dose-dependent manner. These results suggested that in vitro raised plants of C. thwaitesii are an excellent source of antioxidant compounds to be exploited on an industrial level as food additive.

Evaluation of Chemical Composition and In-vitro Biological Activities of Three Endemic Hypericum Species from Anatolia (H. thymbrifolium, H. spectabile and H. pseudolaeve).

In the present work we carried out a phytochemical and biological investigation on three endemic Hypericum species, i.e.Hypericum thymbrifolium (H. thymbrifolium), Hypericum spectabile (H. spectabile) and Hypericum pseudolaeve (H. pseudolaeve) from Anatolia in order to discover new sources of natural compounds for the treatment of inflammatory and neurodegenerative disorders. HPLC-DAD analysis indicated that two naphthodianthrones (pseudohypericin and hypericin) together with chlorogenic acid, rutin, hyperoside, isoquercitrin, kaempferol, quercitrin, quercetin, amentoflavone, and hyperforin are the main compounds present in the methanol extracts. After chemical characterization, all extracts were in-vitro biologically assayed for antioxidant potential by lipid peroxidation inhibitory activity, DPPH, FRAP assays, and superoxide radical scavenging activity, for AChE inhibitory activity by Ellman's method, for COX inhibitory activity by using enzyme immunoassay (EIA) kit, for cytotoxic activity on HeLa and NRK-52E cell lines by MTT assay. The superoxide radical scavenging activity and lipid peroxidation inhibitory activity of H. spectabile (EC50 = 0.430 mg/mL) were more remarkable than that of H. thymbrifolium and H. pseudolaeve. The extracts showed moderate inhibitory activity on AChE (from 49.37% to 63.41%). The best inhibitory activity against COX-1 (71.77% and 77.04%, respectively) and COX-2 ( 64.14% and 72.23%, respectively) were shown by H. thymbrifolium and H. spectabile, which may be due to their richest chlorogenic acid content (0.29576% and 0.23567%, respectively). Cytotoxicity screening results showed that the extracts did not demonstrate significant cytotoxic activity. It was concluded that the most promising extract with antioxidant, anti-inflammatory, and AChE inhibition potential is H. spectabile.