Highest ABTS Radical Scavenging Activity Research Articles

Antioxidative and Anti-Atopic Dermatitis Effects of Peptides Derived from Hydrolyzed Sebastes schlegelii Tail By-Products.

Atopic dermatitis (AD) is a chronic inflammatory skin disorder associated with significant morbidity, including pruritus, recurrent skin lesions, and immune dysregulation. This study aimed to investigate the antioxidative and anti-AD effects of peptides derived from hydrolyzed Sebastes schlegelii (Korea rockfish) tail by-products. Hydrolysates were prepared using various enzymes, including Alcalase, Flavourzyme, Neutrase, and Protamex. Among them, Protamex hydrolysates demonstrated the highest ABTS radical scavenging activity with an RC50 value of 69.69 ± 0.41 µg/mL. Peptides were further isolated from the Protamex hydrolysate using dialysis, fast protein liquid chromatography (FPLC), and high-performance liquid chromatography (HPLC). The most active peptide, STPO-B-II, exhibited a single peak and was identified as a sequence of Glu-Leu-Ala-Lys-Thr-Trp-His-Asp-Met-Lys, designated as MP003. In vivo experiments were conducted using a 2,4-dinitrochlorbenzene (DNCB)-induced AD model in NC/Nga mice. The isolated peptide, MP003, showed significantly reduced AD symptoms, including erythema, lichenification, and collagen deposition. Additionally, MP003 decreased epidermal and dermal thickness, eosinophil, and mast cell infiltration and downregulated the expression of pro-inflammatory cytokines IL-1β, IL-6, and IgE in serum and skin tissues. These findings suggest that peptides derived from Sebastes schlegelii tail by-products may serve as potential therapeutic agents for AD.

Formulation of a Three-Component Essential Oil Mixture from Lavandula dentata, Rosmarinus officinalis, and Myrtus communis for Improved Antioxidant Activity.

The optimization of existing natural antioxidants that are highly effective is crucial for advancements in medicine and the food industry. Due to growing concerns regarding the safety of synthetic antioxidants, researchers are increasingly focusing on natural sources, particularly essential oils (EOs). Combining EOs might enhance antioxidant activity due to increased chemical diversity. This study investigates, for the first time, the antioxidant properties of EOs from Lavandula dentata, Rosmarinus officinalis, and Myrtus communis, both individually and in combination, using the augmented-simplex design methodology. The in vitro evaluation of the antioxidant activity was performed using DPPH and ABTS radical scavenging assays. Chromatography gas-mass spectrometry (CG-MS) revealed that 1,8-cineol (37.27%) and pinocarveol (12.67%) are the primary components of L. dentata; verbenone (16.90%), camphor (15.00%), and camphene (11.03%) are predominant in R. officinalis; while cineol (43.32%) is the main component of M. communis. The EOs showed varying scavenging activities against ABTS and DPPH radicals, with DPPH assay values ranging from 194.10 ± 3.01 to 541.19 ± 3.72 µg/mL and ABTS assay values ranging from 134.07 ± 1.70 to 663.42 ± 2.99 µg/mL. These activities were enhanced when the EOs were combined. The optimal antioxidant blend for DPPHIC50 consisted of 20% L. dentata, 50% R. officinalis, and 30% M. communis. For the highest ABTS radical scavenging activity, the best combination was 18% L. dentata, 43% R. officinalis, and 40% M. communis. These results highlight the potential of EO combinations as new natural formulations for use in cosmeceutical, food, and pharmaceutical sectors.

Biological Activity Evaluation of Olive, Grape, and Fig at Various Mixing Ratios

Purpose: This study aimed to evaluate the antioxidation and enzymatic activity-related qualities of four mixtures prepared with different mixing ratios of olives (O), grapes (V), and figs (F), which are representative ingredients of the Mediterranean diet. Methods: Following mixtures were prepared: olives–figs (OF), grapes–figs (VF), and olives–grapes–figs (OVF). For mixture preparation, the ingredients were mixed in following ratios: olives-to-grapes, olives-to-figs, and grapes-to-figs at ratios of 1:1 (30 g of each) as well as olives-to-grapes-to-figs at a ratio of 1:1:1 (20 g of each). The antioxidant (total polyphenol and flavonoid contents along with 2,2-diphenyl-1-picrylhydrazyl [DPPH] and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) [ABTS] radical scavenging) and enzymatic (α-glucosidase inhibitory and angiotensin-converting enzyme [ACE] inhibitory) activities of the four aforementioned mixtures were measured. Results: OV and OVF mixtures showed the highest total polyphenol (27.25±0.22 mg TAE/g) and flavonoid contents (27.9 ± 2.14 mg QE/g), respectively. DPPH radical scavenging ability was highest in OV (92.86 ± 0.17%). The highest ABTS radical scavenging activity was registered in the OVF mixture (42.88±1.19%). α-glucosidase and ACE inhibitory activity measurements were highest in the OVF mixture (79.75±0.15% and 88.61±0.08%, respectively). Conclusion: Highest physiological activity levels (i.e., anti-oxidation and enzymatic activity) were observed in the olive, grape, and fig (1:1:1) mixture, hence, its regular consumption could improve metabolic syndrome.

Cystoseira myrica: from beach-cast seaweed to fucoidan with antioxidant and anticoagulant capacity

This study highlights the potential of the brown algae Cystoseira myrica, collected from raw beach seaweed wastes, as a reliable source of bioactive fucoidan. Fucoidans are natural bioactive sulfated polysaccharides that are gaining popularity worldwide due to their diverse biochemical composition, attractive physical properties, and functional pharmacological activities. The aim of this work is to valorize the beach-Accumulated brown algae C. myrica by fucoidan extraction and to investigate its anticoagulant and antioxidant activity. Fucoidan was extracted using several steps of papain digestion followed by cetyltrimethylammonium bromide (CTAB) precipitation and calcium chloride treatment to avoid the coextraction of other polysaccharides. Structural features of the extracted fucoidan were investigated by Fourier transform infrared spectroscopy (FT-IR), solid-state nuclear magnetic resonance (NMR) and high-resolution mass spectroscopy. Agarose gel electrophoresis was used to confirm the purity of the isolated fucoidan from C. myrica. Anticoagulant properties were studied in vitro by activated partial thromboplastin time (aPTT) and prothrombin time (PT) assays. Furthermore, the antioxidant activity was investigated by 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and Fe chelating ability. Our results indicate that the approach used was effective in extracting fucoidan with a yield of 3.07%, a high amount of sulfate (27.79%), and fucose was found to be the major monosaccharide component. The extracted fucoidan showed an interesting anticoagulant activity. It prolonged aPTT significantly at a concentration of 10 µg/ml and prolonged PT at high doses. This demonstrated that fucoidan from C. myrica may affect intrinsic pathways while having little impact on the extrinsic mechanism of coagulation. However, the extracted fucoidan significantly exhibited an interesting antioxidant capacity as shown by the higher ABTS radical scavenging activity and Ferrous ion-chelating effect. The current findings suggest that fucoidan isolated from C. myrica has unique structural, antioxidant and anticoagulant properties and offers innovative therapeutic possibilities.

Chemical profile, antioxidant and antimicrobial activity of Pinus heldreichii Christ. Distributed in Bulgaria

Pinus heldreichii Christ. (Bosnian pine), a Tertiary relict and Balkan sub-endemic, has not been comprehensively studied for its essential oil (EO) profile and bioactivity of its different plant parts. This study aimed to determine the EO yield, composition and antimicrobial activity from different parts of P. heldreichii at three different populations (mountains) in Bulgaria. Furthermore, the study assessed the antioxidant activities of plant tissue, including leaves (needles), twigs wood, male and female cones. The EOs yield from different plant parts ranged from 0.09 % (leaves) to 0.74 % (wood of twigs), with monoterpenes being the predominant class. Limonene, α-pinene, β-caryophyllene, germacrene D, β-pinene, and β-myrcene were detected in the EO extracted from all analyzed trees. However, these compounds were not found in the EO extracted from all plant parts of the same trees. Four chemical groups (chemotypes) were identified for EO from twigs, and three chemotypes were identified for EO from leaves. The chemotypes were based on the percent ratio of the main EO constituents (>5 %). Leaves tissue showed the highest values in terms of polyphenols and flavonoids, as well as higher ABTS radical scavenging activity, while the highest antimicrobial activity against Staphylococcus aureus subsp. aureus was seen in the EOs obtained from twigs. This is the first study to identify several chemotypes based on leaf and twigs EO of P. heldreichii distributed in Bulgarian flora. Furthermore, the EO of twigs tips (TT), male cones (MC), and wood of one-two-year-old twigs (WT) of the same trees were reported for the first time. The total polyphenol, flavonoid content, and radical scavenging activity of tissues of annual twigs wood and biennial twigs wood, leaf tissue, MC tissue, and the twigs tips tissue is also reported for the first time in the accessible literature. These findings highlight the potential of P. heldreichii to provide EOs with varying compositions and bioactivities, making them suitable for nutraceutical, pharmacological, and potentially food additive applications. Furthermore, the identification of chemotype accessions in this study suggests their selection for the development of new forest crop as a source for natural products with desirable composition and bioactivity.

Ultrafiltration fractionation of rice protein hydrolysates: Physicochemical properties and potential biological activities of different rice protein hydrolysate fractions

In this study, rice protein hydrolysate (RPH) was separated by membrane ultrafiltration, and three ultrafiltration fractions (UFs) with distinct molecular weights (RPP-1, RPP-2 and RPP-3) were yielded. The physicochemical properties and in vitro bioactivities of these fractions were characterized. Among RPH and its UFs, RPP-1 exhibited highest DPPH radical scavenging activity (IC50 = 4.42 ± 0.08 mg/mL) and ferric reducing antioxidant power (84.21 mg GSH/g). RPP-3 exhibited the highest ABTS radical scavenging activity (IC50 = 0.97 ± 0.01 mg/mL). All the samples exhibited angiotensin converting enzyme inhibitory activity (5.38% ± 0.76%–54.14% ± 3.04%) and α-glucosidase inhibitory activity (−38.90% ± 3.20%–99.99% ± 0.01%). In addition, the activation of alcohol dehydrogenase by RPH and its UFs was observed in this study, with an activation percentage of 46.20% ± 4.96%–199.77% ± 7.71%. However, no inhibitory activity towards acetylcholinesterase was detected. In present study, the potential antialcohol and hepatoprotective activity of RPH was observed. Overall, this study would provide a new idea for the high-value utilization of RPH and a theoretical foundation for the development of bioactivity peptides derived from rice protein.

Comparison of in vitro antioxidant capacities of Phragmites communis Trin. and Phragmites japonica Steud.

The rhizome of Phragmites communis Trin. is used for vomiting and belching by clearing stomach and the sprout is used as tea. Phragmites japonica is similar with P. communis except the color of sheath is purple. This study is aimed to compare the in vitro antioxidant activity, total polyphenol and flavonoids contents of P. communis and P. japonica. The antioxidant activities of fractions from the two Phragmites plants were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assay. The antioxidant activity varied with plant parts and extract solvents. The fractions of leaf extract from the two Phragmites plants (4.06± 1.32-16.47±1.28%) showed higher antioxidant activity by DPPH assay compared with rhizome fractions of two Phragmites plants (0.00±0.00-14.15±0.07%), these are lower compared with ascorbic acid and butylhydroxyanisole (BHA). The highest ABTS radical scavenging activity was found for rhizome ether fraction, namely 74.95±0.56% and 73.04±1.85% for P. communis and P. japonica, these are higher than BHA. The total polyphenol and flavonoids contents were different with plant parts and extract solvents, likewise antioxidant activity. A significant correlation was shown between DPPH and ABTS radical scavenging activity. Considering the results of this study, the leaves and stems of P. communis and P. japonica are expected to be used as natural antioxidants.

Functional modification of thermostable alkaline protease from Bacillus halodurans SE5 for efficient production of antioxidative and ACE-inhibitory peptides from sericin

In this work, the hydrolysis efficiency of recombinant protease_SE5 (r_SE5) on sericin was improved by protein engineering. The S166D variant, obtained from the first round of site-directed mutagenesis, exhibited a 1.4-fold increase in protease activity (602 U/mg) and sericin hydrolysis efficiency (123 μg peptide/mL). Further substitution of Ser166, located on a low-affinity calcium-binding loop, by Glu consequently improved sericin hydrolysis capability. The S166E variant showed remarkable protease activity (671 U/mg) and sericin hydrolysis efficiency (140 μg peptide/mL) with no changes in thermostability compared to r_SE5. Interestingly, the purified S166E efficiently hydrolyzed sericin, with a maximal peptide yield of 540 μg/mL within 60 min. The S166E-derived sericin peptide also exhibited higher ABTS radical scavenging activity (6470 μmol TEAC/L) and angiotensin-I-converting enzyme (ACE) inhibition activity (40%). Additionally, the identification of peptides by LC-MS/MS revealed that two smaller peptides (HHSGVNR and GWWWNSD) with a higher proportion of key amino acid residues were produced by S166E. These peptides were the potential bioactive peptides predicted by the BIOPEP database. Therefore, this study demonstrates the structure–function relevance between the low-affinity calcium-binding loop and hydrolysis of sericin by protease_SE5. S166E is a more powerful protease for producing value-added sericin hydrolysate, which can be used as an active ingredient for food and pharmaceutical applications.

Differentiation of protein types extracted from tilapia byproducts by FTIR spectroscopy combined with chemometric analysis and their antioxidant protein hydrolysates

This research aimed to characterize protein types including sarcoplasmic protein (SP), myofibrillar protein (MP), and alkali-aided protein extract (AP) prepared from tilapia byproducts using water, 0.6 M NaCl, and alkaline solution (pH 11), respectively compared to freeze-dried minced tilapia muscle (CONTROL). Principal component analysis was performed from second derivative FTIR spectra to differentiate protein type. The AP mostly contained β-sheet structure and had low total sulfhydryl content and surface hydrophobicity. SP can be distinguished from MP by the loading plots of the FTIR bands representing the α-helical structure. While the bands for lipids and β-sheet of protein were noted for differentiating AP from CONTROL. After being hydrolyzed by Protease G6, the AP hydrolysate disclosed the highest ABTS radical scavenging activity, while the SP hydrolysate revealed the strongest metal chelating ability. Thus, an understanding of how fish processing waste can be utilized in the production of antioxidant protein hydrolysates has been achieved.

Chemical diversity of wild fennel essential oils (Foeniculum vulgare Mill.): A source of antimicrobial and antioxidant activities

This study aims to provide useful information for the evaluation of wild fennel in terms of their nutritional potential and medicinal values. It is therefore important to bio-prospect the variability of molecules production and the biological properties according to plant georaphical origin. Essential oils (EOs) obtained from leaves of 12 Foeniculum vulgare Mill. populations, growing wild in Tunisia, were analyzed by GC-MS. In addition, their antimicrobial activity was evaluated against eight pathogenic bacteria and fungi. The antioxidant potential of tested oils was evaluated by four different tests: Total antioxidant activity (TAA), DPPH, ABTS and reducing power assay (RPA). In addition, relationships between studied EOs by using Hiearchical Clustering Analysis (HCA) based on Euclidean distance matrix and the multivariate Principal Component Analysis (PCA) were provided. All EOs were found to be rich in monoterpenes hydrocarbons (41.56-63.13%) and phenylpropanoids (35.87-55.29%). Moreover, Limonene (8.40-50.34%) and Methyl Chavicol (35.87%-54.28%) were the main compounds. EOs from Zaghouan displayed the best antioxidant activity (8.5±0.05 mg GAE/g DW). While, EOs from Oued Zarga exhibited the highest DPPH radical scavenging activity (IC50 = 7.7±0.43 mg/mL), and those from Ain Ghlel and Badar regions showed the highest ABTS radical scavenging activity (IC50 = 86.86±0.23 µg/mL and 87.33±0.17 µg/mL, respectively). The most important reducing power assay was found in Ain Ghlel and Charchara EOs (EC50 of 70.46±0.17 and 71.31±0.61µg/mL respectively). Results displayed the sensitivity of the tested strains to fennel EOs especially against Gram-positive strain, and EOs from Zaghouan population exhibited the highest effect against Micrococcus luteus and Staphylococcus aureus. Overall, these results can extend our knowledge of the chemical diversity of wild fennel among geographical origin and pedoclimatic conditions indicating that Tunisian natural populations of F. vulgare had different chemotypes of EOs. In this way, the current study will provide new data and encourage the use of this plant as food. Wild fennel leaf EOs were rich in phenylpropanoids and endowed with significant antibacterial and antioxidant potential, which opens a new prospect for industrial and agronomical exploitation of this species.

Evaluation of Anticholinergic, Antidiabetic and Antioxidant Activity of Astragalus dumanii, an Endemic Plant

The research was conducted to separately evaluate and detect the possible in vitro antioxidant, antimicrobial activity of ethanol extracts prepared from aerial parts and roots of Astragalus dumanii and anti-cholinesterase and α-glucosidase inhibitory activity from only aerial parts of its The antioxidant capacity was tested by scavenging of DPPH and ABTS free radicals. Compared with the standard antioxidant compound gallic acid; Root and aerial part extract showed lower DPPH radical scavenging activity, however aerial part extract demonstrated higher ABTS radical scavenging activity. The phenolic contents were detected as 5.31 ± 0.03 and 13.23 ± 0.05 mg gallic acid equivalent g-1 extract, flavonoid contents were found as 8.26 ± 0.004 and 7.93 ± 0.005 mg Qercetin equivalent g-1 extract. In addition, the effects of the extracts obtained from aerial parts of the plant on acetylcholinesterase, butyrylcholinesterase and a-glycosidase enzymes were investigated in vitro and IC50 values were obtained as 1.47, 0.83 and 0.48 µg mL-1, respectively. When these values were compared with standard substances, it was seen that Astragalus dumanii could be a good enzyme inhibitory agent. Antimicrobial activity of the plant extracts were determined using the microdilution method and the extracts was not observed to have any antimicrobial activities..

Comparative study on structural, biological and functional activities of hydrolysates from Adzuki bean (Vigna angularis) and mung bean (Vigna radiata) protein concentrates using Alcalase and Flavourzyme

The physicochemical features of mung bean protein (MBP) and adzuki bean protein (ABP) hydrolysates derived from Alcalase (MBPHA, ABPHA) and Flavourzyme (MBPHF, ABPHF) were assessed using FTIR, hydrophobicity, emulsion activity, zeta potential, and health-promoting activities. The results proved that the choice of peptidase and substrate both have a significant effect on the hydrolysates in different physicochemical, structural and functional properties. Size exclusion-HPLC was used to fractionate the MBP and ABP hydrolysates. The results demonstrated that Alcalase hydrolysates included smaller peptides than Flavourzyme hydrolysates, and the chromatogram patterns of the two peptidases were similar. The peptides with the most potent antioxidant and ACE-inhibitory properties were identified using MALDI-TOF-MS. The fraction (F4) of MBPHA exhibited the highest levels of metal chelating activity. The Flavourzyme hydrolysates fraction (F2) and the ABPHA fraction (F2) showed the highest ABTS radical scavenging activity and ACE-inhibitory activity, respectively. Pro-Pro was identified in peptide sequences with ABTS radical scavenging activity as an active component while Pro-Gln was identified in peptide sequences with ACE-inhibitory activity. As a result, Pro-Pro and Pro-Gln, respectively, are likely-one of the characteristics of antioxidant and ACE-inhibitory peptides from MBP and ABP. Compared to mung bean and adzuki bean protein as substrate, Alcalase and Flavourzyme as peptidases significant impacted the development of distinct functionalities and biological activities.

Extracts of sorghum bran, grape seed, and green tea: Chromatographic comparison of phenolic profiles and mitigation effect on acrylamide in antioxidant-fortified bread

Phenolic compounds composition of sorghum bran extract (SBE), grapeseed extract (GSE), and green tea extract (GTE) was investigated using a UPLC-DAD-ESI-Q-TOF-MS/MS system. Antioxidant capacity and acrylamide formation in the extract-fortified bread were evaluated through a straight-dough bread model with different levels of the extracts (0.5, 1.0, and 1.5%). All the breads containing 1.5% extracts exhibited significantly higher ABTS radical scavenging activity (17.9 mmol TE kg−1 in GTE bread, 8.9 mmol TE kg−1 in GSE bread, and 8.5 mmol TE kg−1 in SBE bread), compared to the control bread (0.9 mmol TE kg−1). In particular, the enrichment of bread with SBE remarkably reduced the levels of acrylamide by up to 70% without compromising the end-use characteristics. Considering the nutritional convenience and food safety in industrial baking production, the fundamental knowledge acquired through this study may advance the efficient utilization of natural antioxidant extracts for the development of functional bakery products.

Optimization and production of protein hydrolysate containing antioxidant activity from tuna cooking juice concentrate by response surface methodology

To optimize the hydrolysis conditions in the production of antioxidant hydrolysates from tuna cooking juice concentrate (TC) to maximize the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, TC containing 48.91% protein was hydrolyzed with Alcalase 2.4 L, and response surface methodology (RSM) was applied. The optimum hydrolysis conditions included a 2.2% (w/v) Alcalase concentration and 281 min hydrolysis time, resulting in the highest DPPH radical scavenging activity of 66.49% (0.98 ìmol Trolox/mg protein). The analysis of variance for RSM showed that hydrolysis time was an important factor that significantly affected the process (p < 0.05). The effects of different drying methods (freeze drying, hot air drying, and vacuum drying) on the DPPH radical scavenging activity and amino acid (AA) profiles of TC hydrolysate (TCH) were evaluated. Vacuum-dried TCH (VD) exhibited an increase in DPPH radical scavenging activity of 81.28% (1.20 ìmol Trolox/mg protein). The VD samples were further fractionated by ultrafiltration. The AA profiles and antioxidant activities in terms of the DPPH radical scavenging activity, 2,2′-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radical scavenging activity, ferric reducing antioxidant power, and ferrous ion chelating activity were investigated. Glutamic acid, glycine, arginine, and cysteine were the major AAs found in the TCH fractions. The highest DPPH radical scavenging activity was found in the VD-1 fraction (> 5 kDa). The VD-3 fraction (< 10 kDa) exhibited the highest ABTS radical scavenging activity and ferric reducing antioxidant power. The ferrous ion chelating activity was the highest in VD-1 and VD-2 (5 to 10 kDa). In conclusion, this study provided the optimal conditions to obtain high antioxidant activities through TCH production, and these conditions could provide a basis for the future application of TCH as a functional food ingredient.

Characterization of Antioxidant Activity of Heated Mycosporine-like Amino Acids from Red Alga Dulse Palmaria palmata in Japan.

We recently demonstrated the monthly variation and antioxidant activity of mycosporine-like amino acids (MAAs) from red alga dulse in Japan. The antioxidant activity of MAAs in acidic conditions was low compared to that in neutral and alkali conditions, but we found strong antioxidant activity from the heated crude MAA fraction in acidic conditions. In this study, we identified and characterized the key compounds involved in the antioxidant activity of this fraction. We first isolated two MAAs, palythine, and porphyra-334, from the fraction and evaluated the activities of the two MAAs when heated. MAAs possess absorption maxima at around 330 nm, while the heated MAAs lost this absorption. The heated MAAs showed a high ABTS radical scavenging activity at pH 5.8–8.0. We then determined the structure of heated palythine via ESI-MS and NMR analyses and speculated about the putative antioxidant mechanism. Finally, a suitable production condition of the heated compounds was determined at 120 °C for 30 min at pH 8.0. We revealed compounds from red algae with antioxidant activities at a wide range of pH values, and this information will be useful for the functional processing of food.

Melissa officinalis extracts obtained using maceration, ultrasoundand microwave-assisted extractions: Chemical composition, antioxidant capacity, and physical characteristics

Even though Melissa officinalis L. (lemon balm, fam. Lamiaceae) is a well-known medicinal, aromatic, and spicy plant, and its physicochemical profile and biological activity have been investigated, there is no detailed research regarding the influence of solvent nature (polarity) on the extraction of active compounds (total polyphenols and total flavonoids) and antioxidant activities of the obtained extracts. Therefore, this study aimed to evaluate the polyphenol and flavonoid contents, as well as the antioxidant potential of lemon balm extracts obtained by varying the polarity of the extraction solvents (methyl alcohol, acetone, ethyl acetate, and deionized water) and using three extraction techniques (maceration-MAC, ultrasound-UAE, and microwave-assisted extraction-MAE). Two in vitro tests (ABTS and FRAP assays) were used to determine the antioxidant activity of the extracts. Total tannin and total protein contents, extraction yield, and physical properties of the selected extracts were measured as well. The highest content of polyphenols was found for the methanolic and water extracts obtained by all three extraction techniques, while the highest flavonoid yield was detected only in the mentioned methanolic extracts. The microwave reactor provided methanolic, ethyl acetate, and water extracts with the highest ABTS radical scavenging activity, while in the case of UAE, it was methanolic and water extracts, and in the case of MAC only water extracts. On the other hand, among lemon balm extracts from MAC, water extract possessed the highest ferric reducing power, whereas in UAE and MAE, it was ethyl acetate extract. Total tannin content determined in selected water extracts was 2.55, 4.53, and 1.83 mg tannic acid equivalent (TAE)/mL, while total proteins amounted to 1.34, 1.50, and 1.31 mg/mL using MAC, UAE, and MAE, respectively. The content of total extractive substances in the form of the extraction yield was also determined for selected water extracts and amounted to 12.6 % for MAC, 17.2 % for UAE, and 36.8 % for MAE. Further, this research has included the investigation of some physical properties of lemon balm water extracts, such as conductivity (3.68-4.14 mS/cm), pH (5.99-6.43), density (0.854-0.901 g/mL), surface tension (26.0-31.7 mN/m), and viscosity (1.18-1.21 mPa·s). This research represents the base for the future encapsulation of lemon balm extracts, enriched in polyphenol content, in a novel type of biofunctional carrier that potentially can be applied in the pharmacy, chemical industry, and biotechnics.

In vitro ANTI-INFLAMMATORY AND ANTIOXIDANT EVALUATION OF AN INDIGENOUS MEDICINAL PLANT – Pterospermum rubiginosum

The current study was carried out to determine the antioxidant potential, anti-inflammatory activity, and cellular viability of Pterospermum rubiginosum (PR), a tropical tree endemic to the Western Ghats. The antioxidant activities of the PR bark methanolic (PRME) and aqueous extract (PRAQ) were tested using ABTS as well as superoxide, nitric oxide, and hydroxyl radical assays. Total antioxidant activity was evaluated by adopting the colorimetric method and correlation with their antioxidant activities was derived by Pearson co-efficient analysis. The PRME showed the highest ABTS radical scavenging activity, EC50 (46.09µg/ml) followed by PRAQ (52.08µg/ml). Furthermore, the PRME exhibited the highest scavenging activity against superoxide, nitric oxide, and hydroxyl radicals. The MTT assay results revealed good cellular viability up to a concentration of 100µg/ml with an EC50 (106.869µg/ml). The inflammatory mediators such as Cox-2, IL-1β, IL-6, and NF-kB were reduced during the treatment of PRME in LPS stimulated RAW cells. The stress marker in rat liver cells such as glutathione reductase (GR), glutathione peroxidase (GPx), and reduced glutathione (GSH) levels was found in normal levels when compared to the untreated group of rats. The antioxidant enzyme superoxide dismutase and catalase also exhibited notable bioactivity in PRME treated groups up to a concentration of 1000µg/ml. The present study showed excellent In vitro and In vivo antioxidant activity; the potent anti-inflammatory ability of PRME in reducing the LPS induced inflammation in cell culture conditions.

Comparative study of phytochemicals and bioactivities of the leaf extracts of Curcuma amada and Curcuma karnatakensis

Curcuma species are known for their medicinal properties, and it is essential to evaluate under-explored species for their phytocomposition and their practical applications. The present work evaluated the phytochemical, antioxidant, antimitotic, and antiproliferative properties of aqueous methanolic extract from leaves of Curcuma amada and Curcuma karnatakensis. Quantitative phytochemical assays revealed higher amounts of phenols (11.00 ± 0.29 mg GAE / g DW), flavonoids (8.80 ± 0.03 mg QE / g DW), tannins (3.30 ± 0.02 mg CE / g DW) and saponins (53.23 ± 0.02 mg DE / g DW) in C.amada extracts compared to C. karnatakensis. Aqueous methanolic extract of C. amada exhibited higher radical scavenging activity for DPPH (IC50= 127.70 ± 4.05 μg / mL), Ferrous ion chelation (IC50= 287.83 ± 6.38 μg / mL) and CUPRAC antioxidant potential value of 6.33 ± 0.06 mg Trolox equivalent (TE)/g DW. C. karnatakensis exhibited higher ABTS radical scavenging activity with an IC50 value of 96.97 ± 1.58 μg / mL. Further, GC-MS analysis of extracts identified several sesquiterpenoids in both species having significant biological activities. The two extracts exhibited mild antimitotic and antiproliferative properties against Allium root cells and yeast cells. C. amada leaf extract showed higher anti-inflammatory properties than C. karnatakensis. This is the first comprehensive report on leaf phytocomposition, which could be exploited for pharmacological utilization.

Investigation of Antioxidant Activities of Fruit, Leaf, and Branche Extracts of White (Morus alba L.) and Black (Morus nigra L.) Mulberry Species from Diyarbakır

This study aims to determine the antioxidant activities of fruit, leaf, and branche extracts of Morus alba L. and Morus nigra L. species collected from Diyarbakır. Different in vitro methods such as Total Phenol Content (TPC), Total Flavonoid Content (TFC), Copper (II) Ion Reducing Antioxidant Capacity (CUPRAC), 2,2´-diphenyl-1-pikrilhidrazil (DPPH), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging methods were used to evaluate the antioxidant activities of the two Morus species. The highest total phenolic amount belongs to acetone extract (257.5±0.05 μg gallic acid equivalent mg-1) obtained from M. nigra branches, while the highest total flavonoid content was found in acetone extract (185.8±0.15 μg catechin equivalent mg-1) obtained from M. nigra leaves. In the CUPRAC test, the highest activity was observed in methanol extracts from M. alba branches (IC50 values is 35.63±0.22 μg mg-1). The IC50 values of the DPPH radical scavenging activity of the extracts are between 128.1±0.13 and 2494.7±0.30 μg mg-1, the best radical scavenging activity was demonstrated by acetone extract (128.1±0.13 μg mg-1) prepared from the M. alba branche. The highest ABTS radical scavenging activity belongs to the methanol extract (75.5±0.25 μg mg-1) obtained from the M. alba branches. The results prove that both Morus species possess antioxidant properties, not only in leaves and fruits but also in the branches. At the end of the study, it was determined that M. alba and M. nigra have medicinal usage value which can be used as natural antioxidants.

Characterization of Protein Hydrolysates from Fish Discards and By-Products from the North-West Spain Fishing Fleet as Potential Sources of Bioactive Peptides.

Fish discards and by-products can be transformed into high value-added products such as fish protein hydrolysates (FPH) containing bioactive peptides. Protein hydrolysates were prepared from different parts (whole fish, skin and head) of several discarded species of the North-West Spain fishing fleet using Alcalase. All hydrolysates had moisture and ash contents lower than 10% and 15%, respectively. The fat content of FPH varied between 1.5% and 9.4% and had high protein content (69.8–76.6%). The amino acids profiles of FPH are quite similar and the most abundant amino acids were glutamic and aspartic acids. All FPH exhibited antioxidant activity and those obtained from Atlantic horse mackerel heads presented the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, reducing power and Cu2+ chelating activity. On the other hand, hydrolysates from gurnard heads showed the highest ABTS radical scavenging activity and Fe2+ chelating activity. In what concerns the α-amylase inhibitory activity, the IC50 values recorded for FPH ranged between 5.70 and 84.37 mg/mL for blue whiting heads and whole Atlantic horse mackerel, respectively. α-Glucosidase inhibitory activity of FPH was relatively low but all FPH had high Angiotensin Converting Enzyme (ACE) inhibitory activity. Considering the biological activities, these FPH are potential natural additives for functional foods or nutraceuticals.