Isolation of antioxidative and ACE inhibitory peptides from protein hydrolysate of skipjack (Katsuwana pelamis) roe

Bioactive peptides from protein hydrolysate of defatted skipjack (Katsuwonus pelamis) roe with 5% degree of hydrolysis (DH) prepared by Alcalase digestion were isolated and characterised. Two active fractions with ABTS radical scavenging activity (973.01–1497.53 μmol TE/mg sample) and chelating activity (0.05–0.07 μmol EE/mg sample) from consecutive purification steps including ultrafiltration, cation exchange column chromatography and reverse phase high performance liquid chromatography (RP-HPLC), were subjected to analysis of amino acid sequence by LC–MS/MS. Seven dominant peptides with 6–11 amino acid residues were identified as DWMKGQ, MLVFAV, MCYPAST, FVSACSVAG, LADGVAAPA, YVNDAATLLPR and DLDLRKDLYAN. These peptides were synthesised and analysed for ACE-inhibitory activity and antioxidative activities. MLVFAV exhibited the highest ACE inhibitory activity (IC₅₀ = 3.07 μM) (p < 0.05) with no antioxidative property, whilst DLDLRKDLYAN showed the highest metal chelating activity, ABTS radical and singlet oxygen scavenging activities. Therefore, peptides prepared from skipjack roe could be further employed as a functional food ingredient.     

In vitro antioxidant study of vegetable oils containing conjugated linolenic acid isomers

The study aimed to evaluate the antioxidant activity (in vitro) of vegetable oils containing conjugated linolenic acid (CLnA) isomers such as α-eleostearic and punicic acid and also to evaluate the comparative effectiveness of these vegetable oils due to presence of cis–trans isomers in variable amount. Different in vitro methods were used to evaluate the free radical scavenging activity, hydroxyl radical scavenging activity, metal chelating activity and reducing activity of oils at different concentrations of CLnA isomers such as 125, 250 and 375 μg/mL. Inhibition on lipid peroxidation was evaluated by measuring thiobarbituric acid responsive substance (TBARS) and conjugated diene formation at 125 and 250 μg/mL concentrations of CLnA. Both the oils showed potent free radical scavenging activity at 375 μg/mL concentration. In contrary, these oils showed higher hydroxyl radical scavenging, metal chelation and reducing activity at lower concentration i.e. 125 μg/mL. TBARS formation and conjugated diene formation was lower i.e. inhibition of lipid peroxidation was maximum at 125 μg/mL of both CLnA isomers. Overall, both the oils showed better antioxidant activity at lower concentration due to better oxidative stability and bitter gourd oil showed more prominent antioxidant activity than snake gourd oil due to presence of higher trans content.     

Determination of in vitro antioxidant activity of fennel (Foeniculum vulgare) seed extracts

In this study, the antioxidant activity of water and ethanol extracts of fennel (Foeniculum vulgare) seed (FS) was evaluated by various antioxidant assay, including total antioxidant, free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, metal chelating activities and reducing power. Those various antioxidant activities were compared to standard antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and α-tocopherol. The water and ethanol extracts of FS seeds showed strong antioxidant activity. 100 μg of water and ethanol extracts exhibited 99.1% and 77.5% inhibition of peroxidation in linoleic acid system, respectively, and greater than the same dose of α-tocopherol (36.1%). The both extracts of FS have effective reducing power, free radical scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, and metal chelating activities. This antioxidant property depends on concentration and increasing with increased amount of sample. In addition, total phenolic compounds in the water and ethanol extracts of fennel seeds were determined as gallic acid equivalents. The results obtained in the present study indicated that the fennel (F. vulgare) seed is a potential source of natural antioxidant. Although, the tests presented here show the usefulness of FS extracts as in vitro antioxidants it still needs to be that this extracts show their activity in emulsions, biological systems, health implications or dry foods.     

Antioxidant peptides obtained from goose egg white proteins by enzymatic hydrolysis

In this study, antioxidant peptides from goose egg white proteins produced using various enzymes were purified and characterised. Two peptides were named as p14 and p16, showing the highest scavenging activity of 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical and the highest metal ion chelating activity, respectively. The sequences of p14 and p16 were identified to be STMMEERRMKVY (1560.72 Da) and DVFRELRVQ (1161.62 Da), respectively. The sequence of p14 has a similarity of 75% to ovalbumin from Meleagris gallopavo and the sequence of p16 has a similarity of 67% to ovalbumin from Taeniopygia guttata. IC₅₀ values of p14 and p16 were determined, and results showed that DPPH radical scavenging activity was 81.6 and 205.5 μm , 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulphonicacid)(ABTS) radical scavenging was 88.4 and 153.8 μm , hydroxyl radical scavenging was 85.5 and 116.3 μm and metal ion chelating was 170.6 and 117.9 μm , respectively. The two identified peptides from goose egg white hydrolysates act as potent natural antioxidant agents.     

A novel antioxidant and antimicrobial peptide from hen egg white lysozyme hydrolysates

Hen egg white lysozyme (HEWL) was hydrolyzed with papain, trypsin and a combination of the two to isolate antioxidant peptides. The prepared hydrolysates were evaluated for antioxidant activity using DPPH and ABTS radical scavenging, metal ion chelation and lipid peroxidation inhibition. The obtained hydrolysate by a combination of the two enzymes exhibited the highest antioxidant activity compared to other hydrolysates and elected for isolation of antioxidant peptides by reverse-phase high-performance liquid chromatography (RP-HPLC). A most potent fraction namely F2 fraction, identified to be NTDGSTDYGILQINSR (MW: 1753.98 ± 0.5 Da) using tandem mass spectrometry. The antimicrobial activity of the F2 peptide was tested using radial diffusion assay (RDA). Our results showed that this peptide has inhibitory effects on both Gram-negative and Gram-positive bacteria. Minimum inhibition concentration (MIC) values of the F2 peptide against Escherichia coli and Leuconostoc mesenteroides bacteria were 355.64 (±2.2) and 442.25 (±2.8) μg/ml, respectively.     

Influence of postharvest gamma irradiation treatment on the content of bioactive compounds and antioxidant activity of fenugreek (Trigonella foenum–graceum L.) and spinach (Spinacia oleracea L.) leaves

Fenugreek and spinach leaves after irradiation in the dose range of 0.25–1.5 kGy were evaluated for the content of bioactive compounds and antioxidant activity using DPPH radical scavenging, ferric reducing ability power (FRAP), hydroxyl radical scavenging and ferrous ion chelating assays. Results of the study revealed that bioactive content except total ascorbic acid was significantly (p ≤ 0.05) higher in fenugreek compared to spinach. Data analysis revealed that gamma irradiation treatment significantly (p ≤ 0.05) enhanced the content of individual as well as total bioactive components of both vegetables. Positive correlation (r = 0.92) existed between gamma irradiation and total phenolics. The results of the antioxidant activity as determined by above mentioned assays revealed a significant (p ≤ 0.05) decrease in EC₅₀ values and a corresponding increase in antioxidant content and activity due to irradiation. Comparison of the increase in inhibition percentage, reducing power and chelating efficiency revealed that treatment of irradiation was significantly (p ≤ 0.05) effective in enhancing the ferric reducing power of both the vegetables (3.1–37.5% for fenugreek, 4.1–42.8% for spinach) and OH radical scavenging for spinach (1.5–22.4%) compared to fenugreek (0.78–13.1%). The present investigation suggested that postharvest radiation treatment to fenugreek and spinach has a potential to enhance their antioxidant content and activities, besides acting as a photo-sanitary treatment.Industrial relevanceThe increasing demand of convenience, wholesome and health promoting foods has resulted in search of new technologies to improve the shelf-life and at the same time preserve the nutritional quality. Prolonging postharvest storage, while enhancing the content of bioactive compounds will have a positive impact on both the industry and consumers. The present study demonstrated that postharvest radiation treatment of fenugreek and spinach can be used a novel approach to enhance their bioactive composition and antioxidant activity.     

Purification and identification of antioxidant peptides from corn gluten meal

Corn gluten meal was hydrolyzed by alkaline protease and Flavourzyme to obtain the antioxidant peptides. The antioxidant activities of the hydrolysates or peptides were evaluated by free radical scavenging capacity (1,1-diphenyl-2-picrylhydrazyl/2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt/hydroxyl radical/superoxide radical anion), metal ion (Fe²⁺/Cu²⁺) chelating activity and lipid peroxidation inhibitory capacity. The hydrolysates were separated by ultrafiltration, and those with molecular weight <10 kDa exhibited highest antioxidant activity in all relevant assays. The hydrolysates were subsequently purified by gel filtration chromatography, and fraction F3 showed the highest antioxidant activity. Three peptides were identified from fraction F3 using LC–ESI–Q–TOF MS/MS as Leu-Pro-Phe (375.46 Da), Leu-Leu-Pro-Phe (488.64 Da) and Phe-Leu-Pro-Phe (522.64 Da). These peptides exhibited good free radical scavenging activity and lipid peroxidation inhibitory effect. Thus, corn gluten meal may be used as a potential source of antioxidant peptides for food and nutraceutical applications.     

Antioxidant properties and components of some commercially available varieties of rice bran in Pakistan

The antioxidant activity of five indigenous rice bran varieties, i.e. Rice bran-Super kernel (RB-kr), Rice bran-Super 2000 (RB-s2), Rice bran-Super Basmati (RB-bm), Rice bran-Super-386 (RB-86) and Rice bran-Super fine (RB-sf), collected from the same agricultural plots, was evaluated. The order of antioxidant activity was evaluated by measurement of total phenolic content, antioxidant activity in linoleic acid system, reducing power, metal chelating ability, scavenging capacity by DPPH radical and ABTS cation radical and conjugated dienes. Determination of major antioxidant components reported in rice bran, i.e. tocopherols, tocotrienols and γ-oryzanol, was made using reverse phase HPLC. However, for comparison of tocopherol content, quantification was also done by voltammetry. The overall order of antioxidant activity was RB-kr > RB-s2 > RB-bm > RB-86 > RB-sf. However, according to the chelating activity and conjugated dienes assays the antioxidant efficacy of RB-sf was higher than RB-bm and RB-86. Antioxidant power was correlated with growth period and irrigation water demand by a particular variety. A strong correlation of these two parameters with antioxidant activity was observed. RB-kr has the longest growth period and takes the least amount of water out of the series and exhibits highest antioxidant activity. Strongly reverse behavior was observed in case of RB-sf.     

Identification of bioactive peptides after digestion of human milk and infant formula with pepsin and pancreatin

Seven human milks were subjected to an in vitro digestion with pepsin and pancreatin to identify the peptides released from human proteins. On the basis of their sequences, 11 of the 23 peptides were synthesised and their angiotensin converting enzyme (ACE)-inhibitory and antioxidant activities were measured. The β-casein peptides HLPLP and WSVPQPK showed potent ACE-inhibitory and antioxidant activity, with a protein concentration needed to inhibit 50% ACE activity (IC₅₀) of 21 μm and a Trolox Equivalent Antioxidant Capacity (TEAC) of 1.297 μmol 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) equivs μmol⁻¹ of peptide, respectively. These activities were determined after digestion of eight infant formulas and compared with those found in digested human milk. One of the infant formulas exhibited a low IC₅₀ value (60.11 μg protein mL⁻¹ of reconstituted formula) and a high TEAC value (1.7056 μmol Trolox equivs mg⁻¹ of protein) and was therefore selected to identify the peptides responsible of these activities.     

Antioxidant properties and polyphenolics composition of common hedge hyssop (Gratiola officinalis L.)

The antioxidant potential of Gratiola officinalis was evaluated by the off-line and on-line HPLC/UV/DPPH radical scavenging assays, phytochemical composition was analyzed by LC/MS. On-line method was validated by using reference antioxidants and linear dependence was found between their concentration and radical scavenging peak area. Radical scavenging capacity (RSC) of methanol and acetone extracts expressed in their concentration required to scavenge 50% of DPPH^? was 0.10% and 0.13%, respectively; the RSC in ABTS^?+ assay was 1093 ± 104 and 746 ± 18 μM of trolox equivalents in 1 g, respectively. Good correlation was observed between total amount of phenolic compounds and RSC. Preliminary HPLC/UV/MS analysis revealed that the main compounds possessing antioxidant activity in the extracts might be phenylpropanoid glycosides; UPLC/UV/ESI-QTOF-MS analysis suggested 15 structures: 2,5-dihydroxy-p-benzenediacetic and caffeic acids, apigenin 6,8-di-C-β-d-glucopyranoside (vicenin-2), apigenin 8-C-α-l-arabinoside 6-C-β-d-glucoside, (shaftoside), forsythoside B, arenarioside, verbascoside (acteoside), amioside, quercetin-6-O-(2-O-acetyl-glucopyranosyl)-glucopyranoside, isoverbascoside, quercetin glucuronide, linariifolioside, methoxy luteolin-7-O-(6-O-acetyl-glucopyranosyl)-glucopyranoside, methoxy luteolin-glucuronide and luteolin glucuronide.     

Retention of Physicochemical and Antioxidant Properties of Dehydrated Bael (Aegle Marmelos) and Palmyra (Borassus Flabellifer) Fruit Powders

Present study was carried out to evaluate the effect of various dehydration techniques such as sun drying, solar drying, drying after freezing (Freeze for one hour followed by mechanical drying at 55 °C), vacuum drying and drying using lab scale air oven on proximate composition and retention of antioxidants in different fruit powder prepared from Bael (Aegle marmelos) and Palmyra (Borassus flabellifer). Moisture content, Total Ash, Crude fiber %, Fat %, Crude protein %, total phenolic content,β –Carotene and antioxidant activity were tested. The antioxidant activity was measured based on the ability of fruit extract to scavenge 1, 1- diphenyl-2-picrylhydrazyl (DPPH). Among different drying treatments the highest fat percentage recorded by the solar dried palmyra fruit powder and there is no significant difference (α= 0.05) between sun drying and vacuumed drying. Higher concentration of β -Carotene and total phenolic content were recorded in vacuum dried samples both in bael and Palmyra fruit powders and it significantly different (α= 0.05) from other treatments. The scavenging activity of bael fruit powder in vacuum drying was ranged from 65.36% to 81.33% of the concentration 200 μg/ml to1000 μg/ml and the palmyra fruit powder was recorded 57.32% to 83.25% of the concentration 200 μg/ml to1000 μg/ml. Vacuum dried fruit powders of palmyra and bael were given highest radical scavenging activity and the scavenging activity of palmyra fruit powder is higher than the bael. Therefore vacuum drying can be recommended as the most effective drying method to protect chemical characteristics and retention of antioxidant properties of fruit powders.     

Isolation and characterization of three antioxidant pentapeptides from protein hydrolysate of monkfish (Lophius litulon) muscle

In the current study, an efficient method had been developed to acquire the antioxidant hydrolysate of monkfish muscle protein (MPH) using trypsin by an orthogonal (L₉(3)⁴) test. Under the optimum conditions of enzymolysis time 4 h, enzyme-to-substrate ratio (E/S) 2%, enzymolysis temperature 40 °C and pH 8.0, the DH (Degree of hydrolysis) and hydroxyl radical scavenging activity of MPH reached 19.83 ± 0.82% and 58.05 ± 3.01%, respectively. By using ultrafiltration, gel filtration chromatography and reversed phase high performance liquid chromatography (RP-HPLC), three antioxidant pentapeptides were isolated from MPH, and their amino acid sequences were identified as Glu-Trp-Pro-Ala-Gln (MPH-P1), Phe-Leu-His-Arg-Pro (MPH-P2), and Leu-Met-Gly-Gln-Trp (MPH-P3) with molecular weights of 629.68 Da, 668.80 Da, and 633.77 Da, respectively. MPH-P1, MPH-P2, and MPH-P3 exhibited good scavenging activities on hydroxyl radical (EC₅₀ 0.269, 0.114 and 0.040 mg/ml), DPPH radical (EC₅₀ 2.408, 3.751, and 1.399 mg/ml), and superoxide anion radical (EC₅₀ 0.624, 0.101, and 0.042 mg/ml) in a dose-dependent manner. MPH-P3 was also effective against lipid peroxidation in the model system. The antioxidant activities of MPH-P1, MPH-P2, and MPH-P3 were due to their small sizes and the presence of antioxidant and hydrophobic amino acid residues within their sequences. The results of this study suggested that the protein hydrolysate and/or its isolated peptides might be effectively used as food additives for retarding lipid peroxidation occurring in foodstuffs.     

Effect of ultrafine grinding on hydration and antioxidant properties of wheat bran dietary fiber

Wheat bran dietary fiber (DF) powders was prepared by ultrafine grinding, whose effects were investigated on the composition, hydration and antioxidant properties of the wheat bran DF products. The results showed that ultrafine grinding could effectively pulverize the fiber particles to submicron scale. As particle size decrease, the hydration properties (water holding capacity, water retention capacity and swelling capacity) of wheat bran DF were significantly (p < 0.05) decreased and a redistribution of fiber components from insoluble to soluble fractions was observed. The antioxidant activities of wheat bran and DF before and after grinding were in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion-chelating activity, reducing power and total phenolic content (TPC). Compared with DF before and after grinding, micronized insoluble DF showed increased chelating activity, reducing power and TPC yet decreased DPPH˙ radical scavenging activity. Positive correlations were detected between chelating activity, reducing power and TPC.     

Structure–mechanism relationship of antioxidant and ACE I inhibitory peptides from wheat gluten hydrolysate fractionated by pH

The aims of this study were to assess bioactive properties (ACE inhibition and antioxidant capacity) from wheat gluten hydrolysate peptides fractionated by pH (4.0, 6.0 and 9.0), to determine peptide action mechanism, and to relate it to the secondary structure and functional groups of peptides. Gluten hydrolysate extracts (GHE) were enriched in peptides with medium hydrophobicity and molecular weight (≈ 60% MH and 5.5 kDa, respectively). Gluten peptides inhibited ACE I by uncompetitive mechanism and a direct relationship between α-helix structure and IC50% value was obtained (r = 0.9127). TEAC and cooper chelating activity from GHE 6.5 were the highest and directly correlated with MH peptides. GHE 9.0 had high carotene bleaching inhibition (47.5 ± 0.3%) and reducing power activity (163.1 ± 2.9 mg S₂O₃^2 − equivalent g^− 1 protein), which were directly related to disulfide bonds content of peptides (r = 0.9982 and 0.9216, respectively). pH was a good alternative to select bioactive peptides from wheat gluten hydrolysate.     

Antioxidant and Anti-diabetic Properties of Caryota Urens (Kithul) Flour

Starch extracted from pith of Caryota urens L. (Family: Arecaceae) palm is known as “Kithul flour” and is claimed to have health benefits according to folklore and Ayurveda. Antioxidants are believed to possess numerous health benefits. However, as yet, health benefits of C. urens flour have not been scientifically investigated. Antioxidant properties of C. urens flour were tested using different in vitro assays namely, 2,2-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS⁺), ferric reducing antioxidant power, oxygen radical absorbance capacity and ferrous ion chelating assays. Total Phenolic Content (TPC) and Total Flavonoid Content (TFC) were also evaluated. Anti-diabetic properties were estimated using alpha amylase and alpha glucosidase enzyme inhibition assays. Dried methanolic extracts of both boiled and raw samples were used in all assays. Results of the study showed that C. urens flour possess free radical scavenging activity(raw 0.02± 0.01 and boiled 0.04±0.01 mg trolox equivalent (TE)/ g flour), electron donating reducing power (raw 0.10±0.03 and boiled 0.36±0.11 mg TE/g flour), oxygen radical absorbance capacity (raw 2.29± 0.71 and boiled 192.3 ± 57.71 mg TE/1 g flour) and metal ion chelating capacity (raw 0.03±0.01 and boiled 0.14±0.04 mg EDTA equivalents /g flour) exhibiting its antioxidant potential. TPC (raw 1.1±0.3 and boiled 5.12 ± 1.89 mg GAE/g flour) and TFC (raw 1.65 ± 0.47 and boiled 6.69 ± 1.27 μg QE/g flour) which are said to be contributed to antioxidant activities were also found. In addition, boiled flour which is generally consumed as a food has shown higher antioxidant activity. Caryota urens flour did not contain marked anti-diabetic properties. It is evident from results that Caryota urens flour has moderate antioxidant property which may associate with its traditional health claims.     

Three novel angiotensin I-converting enzyme (ACE) inhibitory peptides from cuttlefish (Sepia officinalis) using digestive proteases

The angiotensin I-converting enzyme (ACE) inhibitory activities of protein hydrolysates prepared from muscle of cuttlefish (Sepia officinalis) by treatment with various digestive proteases were investigated. The most active hydrolysate was obtained with the crude protease extract from the hepatopancreas of cuttlefish (64.47 ± 1.0% at 2 mg of dry weight/ml) with a degree of hydrolysis of 8%. By gel filtration on Sephadex G-25 and RP-HPLC on C18 column, three novel peptides with high ACE-inhibitory activity were purified and their molecular masses and amino acid sequences were determined. The three peptides Val-Tyr-Ala-Pro, Val-Ile-Ile-Phe and Met-Ala-Trp with IC₅₀ values of 6.1, 8.7 and 16.32 μM, respectively, were novel ACE-inhibitory peptides. Lineweaver–Burk plots suggest that the three purified peptides act as non-competitive inhibitors against ACE. These results suggest that some peptides from cuttlefish could be a beneficial ingredient for nutraceuticals against hypertension.     

Release of multifunctional peptides by gastrointestinal digestion of sea cucumber (Isostichopus badionotus)

Sea cucumber is a benthic marine organism distributed worldwide and used as food in several Asian countries. The species Isostichopus badionotus is captured intensively off the Yucatan Peninsula, Mexico. Boiled I. badionotus was subjected to in vitro simulated gastrointestinal digestion using pepsin and a pepsin–Corolase PP^® mixture. ACE-inhibitory and radical scavenging activities, iron reducing capacity and cytotoxic effects against colorectal cancer cells were evaluated in the hydrolysates and their ultrafiltered fractions. ACE-inhibitory activity was potent in fractions containing peptides <3000 Da, an effect augmented with combined action of gastric (pepsin) and intestinal (Corolase PP^®) enzymes (IC₅₀ = 0.038 ± 0.004 mg/mL). Antioxidant activity was exerted by peptides with low and high molecular weights, depending on hydrolysis method. This is the first report of cytotoxic capacity against colorectal HT-29 cells in peptides from sea cucumber. Sea cucumber hydrolysates and ultrafiltered fractions are potential ingredients for development of functional foods.     

Air-dried capsicum annuum var. acuminatum medium and big: Determination of bioactive constituents,antioxidant activity and carbohydrate-hydrolyzing enzymes inhibition

In the present study the total phenols, flavonoids, carotenoids and capsaicinoids content, the in vitro antioxidant and hypoglycemic activities of extracts of air-dried fruits from two Capsicum annuum cultivars were investigated. A different composition between the two cultivars was evidenced. C. annuum var. acuminatum medium extract presented a major content of phenols, carotenoids, capsaicin and dihydrocapsaicin while C. annuum var. acuminatum big extract is characterized by the highest quercetin, luteolin and kaempferol content with 68.0, 87.6 and 29.7 μg/g dried weight, respectively, analyzed by HPLC. Medium pepper showed the highest radical scavenging activity in DPPH assay (IC₅₀ of 85.3 μg/ml) while big pepper have an interesting activity in ABTS assay (IC₅₀ of 16.4 μg/ml) and the highest inhibition of linoleic acid oxidation with an IC₅₀ value of 1.2 μg/ml after 30 min of incubation. A selective inhibitory activity against α-amylase was demonstrated for C. annuum var. acuminatum big lipophilic fraction (IC₅₀ values of 8.7 μg/ml). The obtained results suggest that C. annuum cultivars could be used as valuable flavor with functional properties for foods.     

Proteomic identification of antioxidant peptides from 400 to 2500 Da generated in Spanish dry-cured ham contained in a size-exclusion chromatography fraction

The processing of dry-cured ham results in an intense proteolysis that leads to the generation of peptides of different sizes and composition, which have been shown to be biologically active. In this study, a total of ninety-three peptides mainly derived from actin, β-enolase, myosin heavy chain, and creatine kinase proteins have been identified from a size-exclusion chromatography (SEC) fraction that resulted as antioxidant by using matrix-assisted laser desorption/ionisation time-of-flight (MALDI-ToF) and nESI-ion trap mass spectrometry. Several of the identified peptides have been synthesised and their antioxidant activity tested in vitro by using DPPH radical-scavenging assay and reducing power analysis. The peptide with sequence SNAAC showed the best results with an IC₅₀ of 75.2 μM in DPPH radical-scavenging assay and 205 μM in ferric-reducing antioxidant power analysis, very good when comparing with the positive control 2,6-di-tert-butyl-4-methyl phenol (BHT) that showed an IC₅₀ of 358.5 μM and 90.3 μM, respectively, in the different assays. These results suggest that Spanish dry-cured ham represents an important source of powerful antioxidant peptides which due to their natural characteristics may represent a highly valuable alternative in human health.     

Analysis of antioxidant activity and antioxidant constituents of Chinese toon

Antioxidant activity of the methanol and water extracts of Chinese toon (Toona sinensis) leaf was evaluated using DPPH radical scavenging and lipid peroxidation assays. Contents of four major types of antioxidants including β-carotene, ascorbate, α-tocopherol and phenolics were also quantified. Open column chromatography followed by semi-preparative HPLC were applied to separate phenolic antioxidants whose contents were subsequently determined by HPLC. The methanol extract demonstrated much higher antioxidant activity than the water extract. Contents of β-carotene, ascorbate, α-tocopherol and phenolics were 1.23 μmol g^?1, 34.2 μmol g^?1, 2.40 μmol g^?1 and 872 μmol gallic acid equivalents g^?1, respectively. Six phenols were isolated. Their structures were characterized as 5-O-galloylquinic acid, gallic acid, methyl gallate, β-1,2,6-tri-O-galloyl-d-glucose, quercetin 3-O-β-d-glucopyranoside and quercetin 3-O-(2′′-O-galloyl)-β-d-glucopyranoside, respectively. The results indicate that phenolic compounds are the dominant antioxidants in Chinese toon. The compounds β-1,2,6-tri-O-galloyl-d-glucose and quercetin 3-O-(2′′-O-galloyl)-β-d-glucopyranoside were reported for the first time in Chinese toon.