Antioxidant activity of three edible seaweeds from two areas in South East Asia

Total phenolic content (TPC) and antioxidant activity (AOA) of 50% aqueous methanol extracts of the marine algae, Padina antillarum, Caulerpa racemosa and Kappaphycus alvarezzi were studied. TPC was measured using Folin-Ciocalteu method while 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP), ferrous ion chelating (FIC) assay and beta carotene bleaching (BCB) assay were used to study their AOA. P. antillarum was found to have the highest TPC, 2430±208 mg gallic acid equivalents (GAE) per 100 g dried sample and ascorbic acid equivalent antioxidant capacity (AEAC), 1140±85 mg AA/100 g. C. racemosa and K. alvarezzi displayed lower TPC and AEAC. C. racemosa had 144±22 mg GAE/100 g dried sample of TPC and 14.3±2.0 mg AA/100 g of AEAC, while K. alvarezzi had 115±35 mg/100 g dried sample of TPC and 37.8±16.8 mg AA/100 g of AEAC. In addition, P. antillarum displayed the highest reducing power, 15.7±2.6 mg GAE/g and highest chelating ability. C. racemosa and K. alvarezzi exhibited lower reducing power, 0.737±0.423 mg GAE/g and 0.561±0.269 mg GAE/g, and lower chelating ability. However, the AOA of these three seaweeds as assessed by BCB assay were equally high.     

Assessment of in vitro antioxidant capacity and polyphenolic composition of selected medicinal herbs from Leguminosae family in Peninsular Malaysia

Determination of total phenolic (TPC), flavonoid and anthocyanin contents, and various antioxidant activities (2,2-diphenyl-1-picrylhydrazyl radical scavenging, ferric reducing power, ferrous ion chelating and lipid peroxidation inhibition) of leaves and flowers of Bauhinia kockiana, Caesalpinia pulcherrima and Cassia surattensis were performed in this study. The B. kockiana flower was found to possess the highest TPC (8280 ± 498 mg GAE/100 g), free radical scavenging activity (ascorbic acid equivalents 14,600 ± 2360 mg AA/100 g) and reducing ability (72.4 ± 8.7 mg GAE/g). Rutin and chlorogenic acid were detected in the plants, where the C. pulcherrima leaf contained the highest amount of rutin (669 ± 26 mg/100 g), while minute amounts of chlorogenic acid were detected in C. surattensis leaf (9.13 ± 0.44 mg/100 g). The C. pulcherrima leaf displayed the highest ferrous ion chelating and lipid peroxidation inhibition activities. Positive correlation was observed between TPC and various antioxidant activities.     

Effect of processing on the antioxidant activity of millet grains

Millets are generally dehulled and subjected to a hydrothermal treatment before consumption, thus the hulls can be used as a potential source of antioxidants. Several millet grains, namely kodo, finger (Ravi), finger (local), proso, foxtail, little and pearl millet were studied. Antioxidant activities of phenolic extracts obtained from whole grains, as well as their corresponding dehulled and cooked grains and hulls were studied for their total phenolic content (TPC), radical scavenging capacity, and antioxidant activity in a β-carotene/linoleate emulsion. The phenolics present in whole grains were identified and quantified using HPLC and HPLC/MS and results were expressed as total for each of the phenolic groups. The TPC ranged from 2 to 112 μmol ferulic acid equivalents/g defatted meal. All varieties exhibited effective inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, peroxyl and superoxide radicals. Dehulling and cooking affected the TPC and radical scavenging and antioxidant activities of the grains, depending on the variety. In general, the antioxidant activity of phenolic extracts was in the order of hull > whole grain > dehulled grain > cooked dehulled grain With the exception of the two finger millet varieties, hulls of other millet grains had high TPC, thus demonstrating their superior antioxidant activity. Hydroxybenzoic acids, hydroxycinnamic acids and flavonoids in whole grains were identified as contributors to the observed effects. Therefore, dehulling of grain and hydrothermal treatments affect the phenolic content and antioxidant potential of millet grains.     

Bioprospecting traditional Pakistani medicinal plants for potent antioxidants

Antioxidant potential of four methanol extracts from three selected plant species, namely Salvia nubicola (Lamiaceae), Acer oblongifolium (Aceraceae) and Hedera nepalensis (Araliaceae) was measured using assays in aqueous and lipid systems. Antioxidant activities were investigated in aqueous systems by using DPPH radical-scavenging assay, ABTS radical-scavenging assay and DNA protection assay, while antioxidant activity in a lipid system was determined by using the thiobarbituric acid-reactive substances (TBARS) assay. Additionally, the Folin-Ciocalteu method was used to measure total phenolic content. Methanol extracts of leaves and flowers of S. nubicola showed the highest Trolox equivalent (TE) values in the case of the DPPH assay, 2484 ± 4.9 mmol TE/g extract, as well as total phenolic content, 139 ± 0.2 mg gallic acid equivalents/g extract. Three fractions (A-C) of the methanol extract of S. nubicola leaves and flowers were produced by semi-preparative HPLC. Fraction B was found to be the most active in the DPPH radical-scavenging assay and had the highest total phenol content. HPLC-DAD and LC-MS revealed rosmarinic acid in S. nubicola extracts and chlorogenic acid and rutin in H. nepalensis extracts as the main phenolic antioxidants.     

Profiling of in vitro neurobiological effects and phenolic acids of selected endemic Salvia species

The ethyl acetate and methanol extracts from 16 Salvia L. species were screened for their inhibitory activity against acetylcholinesterase, butyrylcholinesterase, lipoxygenase, and tyrosinase; the enzymes linked to neurodegeneration. Their antioxidant activity was also tested using DPPH radical scavenging, metal-chelation, and ferric-reducing antioxidant power (FRAP) assays. Total flavonoid content of the extracts was determined by AlCl₃ reagent, while HPLC technique was applied for analysis of various phenolic acids in the extracts. The extracts exerted weak cholinesterase and tyrosinase inhibition, and remarkable inhibition against lipoxygenase (13.07 ± 2.73-74.21 ± 5.61%) at 100 μg ml⁻¹. The methanol extracts showed higher antioxidant activity in DPPH radical scavenging and FRAP assays. The extracts were analyzed for their gallic, protocateuchic, p-hydroxy-benzoic, vanillic, caffeic, chlorogenic, syringic, o- and p-coumaric, ferulic, rosmarinic, and tr-cinnamic acid contents and the methanol extract of Salvia ekimiana (153.50 mg 100 g⁻¹) was revealed to be the richest in terms of rosmarinic acid.     

Phenolic acid analysis and antioxidant activity assessment of oil palm (E. guineensis) fruit extracts

Phenolic compounds in oil palm fruit (E. guineensis) were extracted in soluble free (SFP), insoluble-bound (ISBP) and esterified (EFP) forms. The total phenolic content (TPC) of the oil palm fruit extracts was determined using the Folin–Ciocalteu method and found to range from 5.03 to 9.04 g/L per g of dried weight (DW). The antioxidant activities of oil palm phenolic extracts were analysed using free radical scavenging assays and results showed that oil palm phenolic extracts contained antioxidant activities in the order of ISBP > EFP > SFP. Eight different phenolic acids were identified and quantified using a simple reversed-phase high performance liquid chromatography (HPLC) with a diode array detector (DAD) and liquid chromatography/tandem mass spectrometry (LC/MS/MS). Ferulic, p-hydroxybenzoic and p-coumaric acid were the dominant phenolic acids found in oil palm fruit extracts and ranged from 55 to 376 μg/g of DW.     

Determination of antioxidant activities of various extracts and essential oil compositions of Thymus praecox subsp. skorpilii var. skorpilii

The aim of this work is to examine the chemical composition and antioxidant activity of separated essential oils and different solvent extracts of Thymus praecox subsp. skorpilii var. skorpilii (TPS). The ethanol, acetone, methanol, hexane, aqueous extracts and separated essential oils of TPS were assessed for their antioxidant activities. Antioxidant activities were evaluated by reduction of Mo(VI) to Mo(V), reducing power, superoxide scavenging activity, free radical-scavenging activity, metal chelating activity, linoleic acid peroxidation, hydrogen peroxide scavenging activity and peroxide scavenging activity. Essential oils were characterized in total to be 41 components, whereas 9 components were isolated by column chromatography for antioxidant activity. TPS essential oil was found to contain thymol (40.31%) and o-cymene (13.66%) as the major components. The ethanol, methanol and water extracts exerted significant free radical-scavenging activity. The methanol and water extracts displayed highest superoxide scavenging activity. The water extract has the highest total phenolics (6.211 mg gallic acid (GAE)/g DW) and flavonoids (0.809 mg quercetin/g DW).     

Antioxidant and antibacterial activities of Nephelium lappaceum L. extracts

Ether, methanolic and aqueous extracts of lyophilized rambutan (Nephelium lappaceum L.) peels and seeds were evaluated for phenolic contents, antioxidant and antibacterial activities. High amounts of phenolic compounds were found in the peel extracts and the highest content was in the methanolic fraction (542.2 mg/g dry extract). Several potential antioxidant activities, including reducing power, β-carotene bleaching, linoleic peroxidation and free radical scavenging activity, were evaluated. The peel extracts exhibited higher antioxidant activity than the seed extracts in all methods determined (P < 0.05). The methanolic fraction was found to be the most active antioxidant as shown by their 50% DPPH inhibition concentration, 4.94 μg/mL. The results indicated this fraction exhibited greater DPPH radical scavenging activity than BHT and ascorbic acid (0.32 g dry extract/g BHT or ascorbic acid). Antibacterial activity against eight bacterial strains was assessed by disc diffusion and broth macrodilution methods. All peel extracts exhibited antibacterial activity against five pathogenic bacteria. The most sensitive strain, Staphylococcus epidermidis, was inhibited by the methanolic extract (MIC 2.0 mg/mL).     

The hydroxycinnamic acid content of barley and brewers’ spent grain (BSG) and the potential to incorporate phenolic extracts of BSG as antioxidants into fruit beverages

The hydroxycinnamic acid (HA) content of starting barley for brewers’ spent grains (BSG), whole BSG and phenolic extracts from BSG was measured using high performance liquid chromatography (HPLC) and correlated with antioxidant potential. The effect of BSG phenolic extracts on antioxidant activity of fruit beverages was also assessed (using the total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays). The concentration of HA present in barley extract and BSG was in the order of ferulic acid (FA), p-coumaric acid (p-CA) derivatives, FA derivatives, p-CA, caffeic acid (CA) and CA derivatives. Results suggested that brewing and roasting decreased the HA content. Antioxidant activity was significantly (P < 0.05) correlated with caffeic acid (R² = 0.8309) and total HA (R² = 0.3942) concentrations. Addition of extracts to fruit beverages resulted in a significant (P < 0.05) increase in antioxidant activity of cranberry juice, measured by the FRAP assay. In vitro digestion significantly (P < 0.05) reduced TPC, DPPH and FRAP activity of the fruit beverages.     

Total phenolics, flavonoids and antioxidant activity of tropical fruit pineapple

Pineapple has several beneficial properties including antioxidant activity. The fruit of pineapple was extracted with ethyl acetate, methanol and water. The phenolic content of the extracts was determined by Folin-Ciocalteu method and antioxidant activity was assayed through some in vitro models such as antioxidant capacity by phosphomolybdenum, ??-carotene-linoleate, and radical scavenging activity using ??,??-diphenyl-??-picrylhydrazyl (DPPH) method. The phenolic contents of the extracts as caffeic acid equivalents were found to be highest in methanol (51.1%) followed by ethyl acetate (13.8%) and water extract (2.6%). Antioxidant capacity of the extracts as equivalent to ascorbic acid (??mol/g of the extract) was in the order of methanol extract > ethyl acetate extract > water extract. In comparison with butylated hydroxyanisole (BHA), at 100 ppm of concentration, the antioxidant and free radical scavenging activities of the extracts assayed through ??-carotene-linoleate and DPPH method were also found to be highest with methanol extract followed by ethyl acetate and water extracts. The results indicated that the extent of antioxidant activity of the extract is in accordance with the amount of phenolics present in that extract and the pineapple fruit being rich in phenolics may provide a good source of antioxidant.     

Effects of binary solvent extraction system,extraction time and extraction temperature on phenolic antioxidants and antioxidant capacity from mengkudu (Morinda citrifolia)

An investigation into the effects of ethanol concentration (0–100%, v/v), extraction time (20–120 min) and extraction temperature (25–65 °C) on the extraction of phenolic antioxidants from mengkudu (Morinda citrifolia) was performed using a single-factor experiment. Total phenolic content (TPC) and total flavonoid content (TFC) assays were used for determination of phenolic compounds. Antioxidant capacity was evaluated by measuring the scavenging effect on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radicals. Experimental results showed that extraction conditions had significant effect on extraction of phenolic compounds and antioxidant capacities. The optimised conditions were 40% ethanol for 80 min at 65 °C, with values of 919.95 mg GAE/100 g DW for TPC, 472.73 mg CE/100 g DW for TFC, 791.71 μmol TEAC/100 g DW for ABTS and 1928.5 μmol TEAC/100 g DW for DPPH. TPC was significantly correlated with DPPH under the effects of ethanol concentration (r = 0.932) and extraction time (r = −0.938).     

Solvent effects on phenolic contents and biological activities of the halophyte Limoniastrum monopetalum leaves

Limoniastrum monopetalum is a traditional medicinal species which leaf infusion exhibits antidysenteric properties against infectious diseases. In this study, ten kinds of leaf extracts were used to examine the effect of extraction solvent system with varying polarities on polyphenol contents and DPPH scavenging activity. Then the superoxide scavenging activity and the reducing power of the most promising solvent extracts were evaluated too. Moreover, the efficiency of the best leaf extract has been investigated against pathogenic bacteria and yeast. Eventually leaf extract was hydrolyzed by acid and the phenolics identified by RP-HPLC. Results showed that phenolic contents and antioxidant activities varied considerably as function of solvent polarity. Leaf extract using pure methanol showed the highest polyphenol content (15.85 mg GAE/g DW). Moreover, antiradical capacities against DPPH and superoxide, and reducing power were maxima in acetone/water (8:2) of leaf extract. However, the latter showed a slight antimicrobial activity against human pathogen strains such as Staphylococcus aureus, Micrococcus luteus and Candida holmii. The HPLC analysis revealed several phenolic compounds in L. monopetalum leaf including vanillic and gallic acids as major phenolics. Our findings identified the appropriate solvent for extracting halophyte phenolics which might provide a rich and novel source of natural antioxidants as food additives replacing synthetic ones in food industry.     

Evaluation of antioxidant,antibacterial and anti-tyrosinase activities of four Macaranga species

The methanolic fresh leaf extracts of Macaranga gigantea, Macaranga pruinosa, Macaranga tanarius and Macaranga triloba were screened for their antioxidant properties (AOP), tyrosinase inhibition and antibacterial activities. Total phenolic content (TPC), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, ferric-ion reducing power (FRAP), ferrous-ion chelating (FIC) and lipid peroxidation inhibition (LPI) activities were used to evaluate the AOP. Modified 3,4-dihydroxy-L-phenylalanine (L-DOPA) method was used to determine tyrosinase inhibition activity, whereas antibacterial activity was determined using the disc-diffusion technique. TPC screening of the same species from different collection sites showed no significant difference between sites. M.triloba showed the highest ascorbic acid equivalent antioxidant activity (AEAC), FRAP and LPI values. M. tanarius, which showed the lowest TPC, AEAC, FRAP and LPI activities, exhibited the best FIC activity. M. pruinosa showed the best tyrosinase inhibition activity, whereas M. triloba showed the best antibacterial activity against Gram-positive bacteria species, with minimal inhibition dosage (MID) values as low as 10 μg/disc.     

Enzyme-enhanced extraction of antioxidant ingredients from red algae Palmaria palmata

The effect of various protease and carbohydrase treatments on the extraction of polyphenols and other antioxidant ingredients from the red algae Palmaria palmata (dulse) was investigated. In addition, the relative contribution of different fractions to the overall antioxidant capacity of the hydrolysate was evaluated. Considerable differences were observed both in total phenolic content (TPC) and antioxidant activities of the hydrolysates evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, oxygen radical absorbance capacity (ORAC) and ferrous ion-chelating ability assays. All the proteases tested had significant enhancing effect on the extraction of polyphenols and other active components compared to carbohydrases and cold water extraction (control). The Umamizyme extract had the highest TPC and consequently exhibited the strongest scavenging capacity against DPPH and peroxyl radicals. Further fractionation of the Umamizyme extract revealed that the crude polyphenol fraction possessed the highest peroxyl radical scavenging activity, whereas the crude polysaccharide fraction was more effective for chelating ferrous ions. The data from this study suggest the potential of protease treatment to improve value-added utilization of dulse extracts as antioxidants in functional foods and nutraceuticals.     

Antioxidant activity of the essential oil and various extracts of Nepeta flavida Hub.-Mor. from Turkey

This study was designed to examine the chemical composition and in vitro antioxidant activity of the essential oil and various extracts (hexane, dichloromethane and methanol sub-fractions) of Nepeta flavida. GC and GC–MS analyses of the essential oil resulted in the identification of 68 compounds, representing 96.4% of the oil; 1,8-cineole (38.9%) and linalool (25.1%) were the main components, comprising 64.0% of the total oil. The samples were subjected to a screening for their possible antioxidant activities by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene-linoleic acid assays. In the first case, the IC₅₀ value of the N. flavida essential oil was determined to be 42.8 ± 2.19 μg/ml. Among the extracts, the strongest activity was exhibited by the polar sub-fraction of the methanol extract with an IC₅₀ value of 63.2 ± 1.75 μg/ml. In the β-carotene-linoleic acid system, N. flavida essential oil exhibited 86.3% ± 1.69 inhibition against linoleic acid oxidation. Among the extracts prepared with various solvents, a correlation was observed between the polarity and antioxidant activity. The extracts exhibited the same activity pattern in this system the most active one is the polar sub-fraction, 79.7% ± 0.89. On the other hand, 1,8-cineole, a major compound of the essential oil, exhibited marked antioxidant activity in both systems, whereas the other compound, linalool, did not show any activity. The amount of total phenolics was highest in the polar and non-polar sub-fractions. Particularly, a positive correlation was observed between the total phenolic content and the antioxidant activity of the extracts. As estimated from the results, amounts of phenolic compounds were less in hexane and dichloromethane extracts than in the others. In conclusion, antioxidant potentials of polar and non-polar methanol sub-fractions could be attributed to their high phenolic contents. In both systems, antioxidant capacities of BHT, ascorbic acid, curcumin and α-tocopherol were also determined in parallel experiments.     

Antioxidant activity and proline content of leaf extracts from Dorystoechas hastata

Dorystoechas hastata (D. hastata) is a monotypic plant endemic to Antalya province of Turkey. D. hastata leaves are used to make a tea locally called “çalba tea”. Diethyl ether (E), ethanol (A), and water (W) were used for the sequential preparation of extracts from dried D. hastata leaves. A hot water extract (S) was also prepared by directly boiling the powdered plant in water. The antioxidant activities of the extracts were tested by ferric thiocyanate (FTC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging methods. E extract exhibited the greatest antioxidant activity with FTC method, whereas S extract exhibited the lowest IC₅₀ value (6.17 ± 0.53 μg/ml) for DPPH radical scavenging activity. Total phenolic contents of the extracts were estimated by Folin–Ciocalteu method and S extract was found to contain the highest amount (554.17 ± 20.83 mg GAE/g extract) of phenolics. Extract A contained highest flavonoid content and there was a inverse linear correlation (R² = 0.926) between IC₅₀ values for DPPH radical scavenging activity and flavonoid contents of all extracts. Reducing power of extracts increased in a concentration-dependent manner. S extract was found to possess higher reducing power than equivalent amount of ascorbic acid at 20 and 25 μg/ml concentrations. Linear correlation between reducing power and concentration of E, A, and W extracts (R² > 0.95) was observed. A, W, and S extracts contained relatively high levels of proline. The results presented suggest that D. hastata may provide a natural source of antioxidants and proline.     

Organic acids,antioxidant capacity,phenolic content and lipid characterisation of Georgia-grown underutilized fruit crops

Four underutilized Georgia-grown fruit crops, namely loquat (Eriobotrya japonica), mayhaw (Crataegus sp.), fig (Ficus carica), and pawpaw (Asimina triloba), and their leaves were analysed for total polyphenols by Folin–Ciocalteau method, and antioxidant capacity by ferric-reducing antioxidant power (FRAP) and Trolox-equivalent antioxidant capacity (TEAC) assays. Organic acids and phenolic compounds were identified by RP-HPLC. For lipid profile, fruits were separated into two fractions – seed and fruit (i.e., without seed); lipid was extracted using the Folch method and analysed for fatty acids, phytosterols, tocopherols, and phospholipids. The major organic acid identified in all samples was malic acid (177–1918 mg/100 g FW). The predominant phenolic acids in all the fruits were gallic (1.5–6.4 mg/100 g FW) and ellagic (0.2–33.8 mg/100 g FW), and the most abundant flavonoid was catechin (12.2–37.8 mg/100 g FW). Total lipid content varied from 0.1% in mayhaw fruit to 21.5% in pawpaw seed. Linoleic acid was the predominant fatty acid in all of the samples (28.2–55.7%).     

Antioxidant properties of various solvent extracts from lychee (Litchi chinenesis Sonn.) flowers

The antioxidant capacities of the acetone, methanol and water extracts of hot-air dried lychee (Litchi chinenesis Sonn.) flowers were estimated with Trolox equivalent antioxidant capacity (TEAC) assay, reducing power and 2,2-diphenyl-2-picrylhydrazyl hydrate (DPPH) radical-scavenging assay. The contents of antioxidant components in these extracts were also determined. Results showed that the highest and lowest contents of these components including phenols, flavonoids and condensed tannins were found in acetone and water extracts, respectively. The antioxidant activities of the lychee flower extracts for all assays were in the order: acetone extract > methanol extract > water extract. The contents of antioxidant components in these extracts were correlated with antioxidant activities.     

Phenolic composition,antioxidant and acetylcholinesterase inhibitory activities of Sclerocarya birrea and Harpephyllum caffrum (Anacardiaceae) extracts

Total phenolic content, proanthocyanidins, gallotannins, flavonoids, and antioxidant activities of Sclerocarya birrea and Harpephyllum caffrum methanolic extracts were evaluated using in vitro assays. S. birrea young stem extract contained the highest levels of total phenolic content (14.15 ± 0.03 mg GAE/g), flavonoids (1.21 ± 0.01 mg CE/g) and gallotannins (0.24 ± 0.00 mg GAE/g). H. caffrum stem bark extract had the highest content of proanthocyanidins (1.47%). The EC₅₀ values of the extracts in the DPPH free radical scavenging assay ranged from 4.26 to 6.92 μg/ml, compared to 6.86 μg/ml for ascorbic acid. A dose-dependent linear curve was obtained for all extracts in the ferric-reducing power assay. Dichloromethane and methanol extracts exhibited dose-dependent acetylcholinesterase inhibitory activity. Similarly, all extracts exhibited high antioxidant activity comparable to butylated hydroxytoluene based on the rate of β-carotene bleaching (84.1–93.9%). The two Anacardiaceae species provide a source of natural antioxidants and acetylcholinesterase inhibitors, and may be beneficial to the health of consumers.     

Antioxidant properties of various solvent extracts of mulberry (Morus indica L.) leaves

The antioxidant properties and total phenolic contents of methanol, acetone and water extracts of mulberry (Morus indica L.) leaves were examined. Various experimental models including iron (III) reducing capacity, total antioxidant capacity, DPPH radical scavenging activity and in vitro inhibition of ferrous sulphate-induced oxidation of lipid system were used for characterization of antioxidant activity of extracts. The three extracts showed varying degrees of efficacy in each assay in a dose-dependent manner. Methanolic extract with the highest amount of total phenolics, was the most potent antioxidant in all the assays used. In addition, the effect of temperature (50 °C and 100 °C), pH (3, 5, 7, 9 and 11) and storage (5 °C) on the antioxidant activity of methanolic extract was investigated. The antioxidant activity of the extract remained unchanged at 50 °C and was maximum at neutral pH. The extract stored at 5 °C in the dark was stable for 30 days after which the antioxidant activity decreased (p ? 0.05) gradually. On the basis of the results obtained, mulberry leaves were found to serve as a potential source of natural antioxidants due to their marked antioxidant activity.