Antioxidant Fractions Research Articles

Tailored recovery of antioxidant fractions enriched in caffeine and phenolic compounds from coffee pulp using ethanol-modified supercritical carbon dioxide

Coffee pulp (CP) is the by-product of coffee processing that urgently needs to be revalorized using sustainable technologies. This work applied a design of experiment (DoE) for modeling the extraction of bioactive compounds from CP using supercritical carbon dioxide (sc-CO2) with ethanol as a co-solvent under variable conditions (temperature, pressure, and ethanol percentage). Considering extraction efficiency (per unit of CP) and extraction selectivity (per unit of extract), results showed that ethanol percentage significantly enhanced the efficiency of total phenolic content, as well as the selectivity of chlorogenic acid and protocatechuic acid (p < 0.05). The effects of temperature and pressure in EtOH-modified sc-CO2 extraction exhibited non-linear patterns, highlighting the interaction between temperature and ethanol. Total phenolic content was responsible for the antioxidant activity of the EtOH-modified sc-CO2 extracts, which was also affected by caffeine in the extracts. The extraction condition that balances both efficiency and selectivity was found at 65 °C, 23 MPa, and 9 % ethanol as co-solvent. The extraction yield (5.8 mg/g CP) was 3.8 times and 6.7 times lower than that of aqueous and ethanolic extraction, respectively, indicating a relatively lower efficiency of sc-CO2. However, the selectivity of caffeine (106.3 mg/g extract) and sum of identified phenolic compounds (16.3 mg/g extract) such as chlorogenic, protocatechuic, and caffeic acids, especially for protocatechuic acid (14.9 mg/g extract), was superior to conventional extraction, resulting in the better ABTS chemical (182.3 mg TE/g extract) and cellular antioxidant (384.9 U/mg extract) activities. The notable enrichment of caffeine and protocatechuic acid, together with the antioxidant potential observed in the EtOH-modified sc-CO2 extracts of CP could be promising for functional food, cosmetic, and pharmaceutical industries.

Stevia By-product Fraction has Antioxidant Capacity and has been Evaluated for its Antihyperglycemic and Antilipid Potential

Introduction/Objective: The ethanolic pretreatment of stevia leaves is considered ecologically favorable and increases its yield, purity level, and sensory profile of sweeteners. We investigated the by-product generated so that we can provide a viable and applicable destination to effectively contribute to the production and industrial chain of stevia sweeteners. Recently, an antioxidant and antidiabetic fraction was obtained through stevia methanolic extraction. However, this process is costly, has a low yield, and is non-green. In this study, we got an antioxidant fraction of stevia from its pretreatment by-product, the ethanolic extract, characterized its bioactive compounds – mainly phenolic acids and flavonoids – and evaluated its antidiabetic and antilipid capacity. Methods: The ethanolic extract was fractionated, then the ethyl acetate fraction was microencapsulated. Physicochemical, Mass Spectrometry (UHPLC/MS-MS), scanning electron microscopy, microencapsulation efficiency, and antioxidant capacity analyses were carried out. The antioxidant capacity was evaluated. Antihyperglycemic capacity was assessed by inhibition of α-glucosidase and α-amylase. For antilipid potential, inhibition of pancreatic lipase was measured. Results: Antioxidant potential was observed mainly in the fraction. Microencapsulation improved the stability of the fraction. Mass spectrometry allowed comparison with phenolic and flavonoid compounds from the methanolic and ethanolic fractions of the by-product. Many compounds were coincident, which may explain the similar antioxidant capacity found. The samples showed more than 90% inhibition of alpha-glucosidase, which may indicate potential antidiabetic activity. All samples showed inhibition of the pancreatic lipase enzyme higher than 80%, reaching 91.05% for the free fraction. Conclusion: This study represented the recovery of a by-product and showed that this antioxidant fraction deserves further investigation for its bioactive potential, especially antioxidant, antidiabetic, and anti-lipid.

Bioactive proteins and antioxidant peptides from Litsea cubeba fruit meal: Preparation, characterization and ameliorating function on high-fat diet-induced NAFLD through regulating lipid metabolism, oxidative stress and inflammatory response

Non-alcoholic fatty liver disease (NAFLD) plays an increasingly significant threat to human health. In this study, the processing by-products of Litsea cubeba fruit meal were defatted by ultrasound-assisted methods, then the acetone-precipitated protein of L. cubeba (LCP) was obtained and structural analysis was performed. LCP was hydrolyzed by a two-step sequential hydrolysis method using alcalase and papain. Subsequently, antioxidant peptide fraction (IV2) was isolated and identified from the resultant hydrolysate through membrane ultrafiltration, Sephadex G-15 chromatography, and liquid chromatograph mass spectrometer (LC-MS). Animal experimentation indicated the potential of IV2 to mitigate hepatic steatosis. Moreover, IV2 could effectively reduce oxidative stress-induced damage by modulating the Keap1-Nrf2 pathway to activate downstream heme oxygenase-1 (HO-1) and NAD(P) H quinone oxidoreductase 1 (NQO1). Integrating metabolomics and transcriptomics revealed enrichment in pathways associated with glycerolipid metabolism and fatty acid β-oxidation, suggesting the principal mechanisms underlying IV2's ameliorative effects on NAFLD. Transcriptome sequencing identified 3092 up-regulated and 3010 down-regulated genes following IV2 treatment. Interaction analyses based on different lipid compositions (DELs) and differentially expressed genes (DEGs) indicated that IV2 primarily alleviated hepatic steatosis by modulating peroxisome proliferator-activated receptor α (PPAR-α) related pathways, thereby augmenting fatty acid β-oxidation within liver cells. These results indicate that IV2 shows potential in improving high-fat diet (HFD)-induced NAFLD, with improved fatty acid β-oxidation and reduced triglyceride biosynthesis emerging as underlying mechanisms.

A Comparison Between Microcapsules of Antioxidant Fraction of Stevia: an Evaluation of Physicochemical Parameters, Stability, Bioaccessibility, and Antidiabetic Capacity

Introduction: The antioxidant fraction of Stevia rebaudiana has important antidiabetic potential, is poorly soluble, and has low stability, which makes its applicability difficult. Materials and Methods: In this study, Stevia's fraction was obtained, characterized by Ultra-high-performance liquid chromatography coupled with mass spectrometry (UHPLCMS/ MS), and microencapsulated with maltodextrin (MDF), gum arabic (GAF), whey protein + maltodextrin (MDWF), and whey protein + gum arabic (GAWF). Then, the following parameters were evaluated: stability and antioxidant capacity over 30 days, physicochemical characteristics, preservation of bioactive compounds by digestion, and in vitro antidiabetic activity. Results and Discussion: The microcapsule with gum arabic showed better microencapsulation efficiency, while its version with maltodextrin provided increased solubility. Microencapsulation reduced the humidity and water activity in the microcapsules, except for the microcapsules containing gum arabic. The combination of maltodextrin and whey protein showed greater luminosity and a whiter powder. The microcapsules with maltodextrin and its mixture with gum arabic kept the phenolic compounds stable for 30 days. The microencapsulation with whey protein increased the antioxidant activity after 15 days of the process and during in vitro digestion. All samples showed 96% inhibition of the α-glucosidase enzyme, and only the microcapsule with maltodextrin inhibited α-amylase (58%). Conclusion: This study showed paramount results so that Antioxidant Stevia Fraction (ASF) can effectively be an adjuvant product to prevent and treat diabetes.

Characterization of the exopolysaccharide produced by Pediococcus acidilactici S1 and its effect on the gel properties of fat substitute meat mince

Exopolysaccharide produced by lactic acid bacteria has various functions. In the present study, one anti-oxidant polysaccharide fraction, namely S1-EPS, was extracted and purified from Pediococcus acidilactici S1, and its structure and its potential effect on the gel properties of fat substitute meat mince were investigated. The results showed that S1-EPS, one of homogeneous polysaccharides, was mainly composed of Gal, Glc, and Man in molar ratio of 7.61: 15.25: 77.13 and molecular weight of 46.975 kDa. The backbone of EPS-S1 contained →2,6)-α-D-Manp-(1→，→2)-α-D-Manp-(1→,→3)-α-D-Glcp-(1 → and a small amount of→6)-β-D-Manp-(1→. The linkages of branches in EPS-S1 were mainly composed of α-D-Manp-(1→ attached to a sugar residue →2,6)-α-D-Manp-(1→O-2 or β-D-Galp-(1→ attached to a sugar residue →2,6)-α-D-Manp-(1→O-6. Furthermore, as S1-EPS increased, the meat minced gel pores decreased, and the surface became smooth. A remarkable inhibitory effect on the lipid oxidation of meat minced gel was found as S1-EPS concentration increased. Overall, S1-EPS was found to have substantial potential in low-fat meat products by serving as a natural, anti-oxidant, and functional additive.

Antioxidant polysaccharide-enriched fractions obtained from olive leaves by ultrasound-assisted extraction with α-amylase inhibition, and antiproliferative activities.

Polysaccharide-rich materials were extracted from the alcohol-insoluble solids of Olea europaea l. **leaves. Structural characteristics were determined by colorimetric techniques, FT-IR, GC-MS, SEC/MALS/VD/DRI, and NMR (1H,13C). The extract and its main macromolecular components were characterized to assess their ability toward antioxidant, α-amylase inhibition, and antiproliferative activities. Results revealed that the ultrasound olive leave extract comprises polysaccharides with uronic acid, galactose, arabinose, and glucose in molar percentages of 11.7%, 11.3%, 7.5%, and 4.9% respectively, constituting 41% of the total mass. In addition, polyphenols (21%) and proteins (9%) are associated with these polysaccharides. Further, the extract showed noticeable ORAC and free radical scavenging abilities, in addition to high in vitro antiproliferative activity against Caco-2 colon carcinoma cell lines. Similarly, the extract exhibited a strong, uncompetitive inhibition of α-amylase by 75% in the presence of the extract with 0.75µg/mL of concentration. This research concludes that ultrasound extraction method can be used for the extraction of polysaccharide-polyphenol-protein complexes. These conjugates exhibit the potential for combined biological activities resulting from a synergistic effect of its compounds, making them promising ingredients for the development of functional food.

Isolation and antioxidant activity of peptides from Chinese hairy tofu.

Hairy tofu is a famous Chinese snack that is made from soybeans and rich in various nutrients. In order to further explore the antioxidant peptides of hairy tofu hydrolysates, seven proteases were used to hydrolyze hairy tofu. The results of in vitro radical scavenging activity showed that hairy tofu hydrolysates obtained by pancreatin exhibited the highest antioxidant activity. After Sephadex G-25 gel filtration and reversed-phase high-performance liquid chromatography (RP-HPLC), 97 peptides were identified in the most antioxidant fraction using liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). Among them, nine peptides were synthesized and their antioxidant activities were assessed using a H2O2-induced oxidative 293T cell model. Finally, four peptides (QCESHK, LAWNEGR, NLQGENEWDQK, and FTEMWR) at concentrations of < 50 μg/ml significantly decreased the malondialdehyde content compared with the model group, displaying in vivo antioxidant activity and low cytotoxicity. Overall, this research provided the choice of using hairy tofu peptides as antioxidant products in the pharmaceutical and food industries.

Antioxidative and Antibacterial Hydro-Ethanolic Fraction from an Asian Edible Mushroom Lentinus sajor-caju (Agaricomycetes) Suppresses Inflammatory Responses by Downregulating COX-2 and iNOS Expression.

Mushrooms are prevalently important sources of pharmaceutically active metabolites. Various mushroom species belonging to the Lentinus genus are recognized for their nutritional and therapeutic properties. One such species is L. sajor-caju, which is renowned in Southeast Asian nations for its culinary value. The primary goal of this study is to investigate the potential medicinal properties of L. sajor-caju, specifically its antioxidant, antibacterial, and anti-inflammatory effects. A hydroethanolic extract was formulated using dried basidiocarps, which exhibited a high phenolic content of approximately 14% and a flavonoid content of approximately 2.7%. The extract demonstrated significant antioxidant potential in in vitro reactions. The extract is sufficiently capable of scavenging free radicals (DPPH and ABTS) and chelate Fe2+ with EC50 values spanning from 186 to 390 μg/mL. In addition, considerable antimicrobial activity against tested pathogenic microorganisms was observed, as indicated by low MIC50 values (256-358 μg/mL). Moreover, the fraction was found to prevent heat-induced protein denaturation which signifies its anti-inflammatory potential. When tested on the RAW 264.7 cell line, reduction in the nitrite production, and downregulation of COX-2 and iNOS mRNA expression was observed which are the key regulator of inflammatory signalling systems. The study, therefore, recommends the use of L. sajor-caju in the medical and pharmaceutical industries for the benefit of humanity.

Biocosmetics Made with Saccharina latissima Fractions from Sustainable Treatment: Physicochemical and Thermorheological Features.

This work deals with the formulation of natural cosmetics enriched with antioxidant fractions from the ultrasound treatment (US) of the brown seaweed Saccharina latissima. The challenge was the development of a cosmetic matrix without jeopardizing the thermorheological features of the creams, adding microparticles containing the antioxidant fractions using two different carriers, mannitol and alginate. The fundamental chemical characteristics of seaweed and the extracts obtained via sonication, as well as the antioxidant properties of the latter, were analyzed. The highest TEAC (Trolox equivalent antioxidant capacity) value was identified for the extracts subjected to the longest processing time using ultrasound-assisted extraction (240 min). A similar yield of microparticle formulation (around 60%) and load capacity (about 85%) were identified with mannitol and alginate as carriers. Color testing of the creams exhibited small total color differences. The rheological results indicated that the testing temperature, from 5 to 45 °C, notably influenced the apparent viscosity of the matrices. All creams were adequately fitted with the two parameters of the Ostwald-de Waele model, with the flow consistency index following an Arrhenius dependency with the testing temperature. Neither hysteresis nor water syneresis was observed in the proposed cosmetics during 6 months of cold storage at 4-6 °C.

Green Extraction of Antioxidant Fractions from Humulus lupulus Varieties and Microparticle Production via Spray-Drying.

The formulation of polymeric microparticles to encapsulate bioactive compounds from two hop varieties (Nugget and Perle) using sequential green extraction processes was performed. The technologies used were ultrasound-assisted extraction (UAE) and pressurized hot water (PHW) extraction. Liquid phases were analyzed for total phenolic content (~2%), antioxidant activity (IC50, DPPH: 3.68 (Nugget); 4.46 (Perle) g/L, TEAC (~4-5%), FRAP (~2-3%), and reducing power (~4%)), protein content (~1%), oligosaccharide content (~45%), and for structural features. The fractions obtained from UAE were selected to continue with the drying process, achieving the maximum yield at 120 °C (Perle) and 130 °C (Nugget) (~77%). Based on these results, the formulation of polymeric microparticles using mannitol as the carrier was performed with these fractions. The production yield (~65%), particle size distribution (Perle: 250-750 µm and Nugget: ~100 µm), and rheological features (30-70 mPa s at 0.1 s-1) were the parameters evaluated. The UAE extracts from hop samples processed using a sustainable aqueous treatment allowed the formulation of microparticles with a suitable yield, and morphological and viscosity properties adequate for potential food and non-food applications.

Liver cirrhosis: physiology, pathology, market analysis, treatments

Azadirachta indica A. Juss is known as the tree of life for its diverse pharmacological applications. This study evaluated the effect of the anxiolytic type of bark of A. indica and mechanisms of action on adult zebrafish. Initially, EtCNeem toxicity was detected against Artemia salina, eliminated with the saponification reaction. The antioxidant potential of the fractions was evaluated. The fraction with higher antioxidant potential was submitted to the preliminary chemical prospecting, FT-IR, and the phenol and flavonoid content was determined. The animals were treated with the most promising antioxidant fraction (1.0; 2.5 or 5.0 mg/mL, i.p.) and underwent acute toxicity, open field tests and Light &amp; Dark Test, evaluating the participation of serotonergic and GABAergic systems in the anxiolytic test. As a result, we selected the fraction of ethyl acetate (F-EtOAc) rich in phenolic and flavonoid compounds with antioxidant potential, which had a sedative and anxiolytic effect on adult zebrafish through serotonergic and GABAergic systems. F-EtOAc prevented alcohol-induced chronic anxiety in adult zebrafish. The results suggest that F-EtOAc is an anxiolytic agent mediated via the GABAergic and serotonergic system

Tandem mass spectrometry (MS/MS) molecular networking guided profiling of small molecules from Aquilaria sinensis (Lour.) Gilg leaves and their bioactivity evaluation.

Agarwood, a fragrant resinous wood mainly formed by Aquilaria spp., is used worldwide as a natural fragrance and traditional medicine. A large amount of Aquilaria sinensis (Lour.) Gilg leaves are underutilised during the process of the agarwood industry, and the development of A. sinensis leaves as tea has recently attracted more and more attention. However, the small molecule profile of A. sinensis leaves and their bioactivities has been rarely reported. To conduct a rapid untargeted liquid chromatography-mass spectrometry (LC-MS) analysis of A. sinensis leaves with a molecular networking (MN) strategy and evaluate its antioxidant and antidiabetic value. A MN-assisted tandem mass spectrometry (MS/MS) analysis strategy was used to investigate the small molecule profile of A. sinensis leaves. Additionally, the integration of antioxidant and α-glucosidase inhibitory assays with MN analysis was executed to expeditiously characterise the bioactive compounds for potential prospective application. Five main chemical groups including phenol C-glycosides, organic acids, 2-(2-phenylethyl) chromones, benzophenone O-glycosides and flavonoids were rapidly revealed from the A. sinensis leaves. Eighty-one compounds were provisionally identified by comparing their MS/MS fragments with canonical pathways. The featured xanthone C-glycosides and benzophenone C-glycosides were recognised as the primary components of A. sinensis leaves. Several dimers and a trimer of mangiferin were reported firstly in A. sinensis leaves. Furthermore, 17 and 14 potential bioactive molecules were rapidly annotated from antioxidant and α-glucosidase inhibitory fraction, respectively. Our findings will help expand the utilisation of A. sinensis leaves and thus promote the high-quality development of agarwood industry.

Whey Protein Hydrolysate Improved the Structure and Function of Myofibrillar Protein in Ground Pork during Repeated Freeze-Thaw Cycles.

Whey protein hydrolysate (WPH) has made a breakthrough in inhibiting oxidative deterioration and improving the quality of meat products during storage. Based on our previous study of extracting the most antioxidant active fraction I (FI, the molecular weight < 1 kDa) from whey protein hydrolysates of different molecular weights, the present study continued to delve into the effects of WPH with fraction I on the structure and function of myofibrillar proteins (MP) in ground pork during the freeze-thaw (F-T) cycles. With the number of F-T cycles raised, the total sulfhydryl content, the relative contents of α-helix, Ca2+-ATPase activity, K+-ATPase activity, solubility, emulsion activity index (EAI), and emulsion stability index (ESI) of MP gradually decreased. Conversely, the carbonyl content and the relative content of random curl showed an increasing trend. In particular, the damage to the structure and the function of MP became more pronounced after three F-T cycles. But, during F-T cycles, FI stabilized the structure of MP. Compared to the control group, the 10% FI group showed a remarkable improvement (p < 0.05) in the total sulfhydryl content, Ca2+-ATPase activity, K+-ATPase activity, solubility, EAI and ESI after multiple F-T cycles, suggesting that 10% FI could effectively inhibit protein oxidation and had the influence of preserving MP function properties. In conclusion, WPH with fraction I can be used as a potential natural antioxidant peptide for maintaining the quality of frozen processed meat products.

Partial Purification and Biochemical Evaluation of Protease Fraction (MA-1) from Mycoleptodonoides aitchisonii and Its Fibrinolytic Effect.

The antioxidative proteolytic fraction, MA-1, was partially purified from Mycoleptodonoides aitchisonii. MA-1 was purified to homogeneity using a two-step procedure, which resulted in an 89-fold increase in specific activity and 42.5% recovery. SDS-PAGE revealed two proteins with a molecular weight of 48 kDa. The zymography results revealed proteolytic activity based on the MA-1 band. MA-1 was found to be stable in the presence of Na+, Ca2+, Fe3+, K+, and Mg2+. MA-1 was also stable in methanol, ethanol, and acetone, and its enzyme activity increased by 15% in SDS. MA-1 was inhibited by ethylenediaminetetra-acetic acid or ethylene glycol tetraacetic acid and exerted the highest specificity for the substrate, MeO-Suc-Arg-Pro-Tyr-pNA, for chymotrypsin. Accordingly, MA-1 belongs to the family of chymotrypsin-like metalloproteins. The optimum temperature was 40 °C and stability was stable in the range of 20 to 35 °C. The optimum pH and stability were pH 5.5 and pH 4-11. MA-1 exhibited stronger fibrinolytic activity than plasmin. MA-1 hydrolyzed the Aα, Bβ, and γ chains of fibrinogen within 2 h. MA-1 exhibited an antithrombotic effect in animal models. MA-1 was devoid of hemorrhagic activity at a dose of 80,000 U/kg. Overall, our results show that M. aitchisonii produces an acid-tolerant and antioxidative chymotrypsin-like fibrinolytic enzyme, and M. aitchisonii containing MA-1 could be a beneficial functional material for the prevention of cardiovascular diseases and possible complications.

Bio-Assay-Guided Isolation of Fractions and Constituents with Antioxidant and Lipid-lowering Activity from Allium cepa.

Active fractions and constituents with antioxidant and lipid-lowering activities were investigated using bio-assay-guided isolation and identification. The data showed that the antioxidant fraction of A. cepa was AC50%, the main constituents of which were quercetin and isoquercitrin, by way of both ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) and bio-assay-guided purification and elucidation. Similarly, the lipid-lowering active fraction of A. cepa was AC30% with the main constituents of 3,4-dihydroxybenzoic acid and quercetin 3,4'-O-diglucoside. Also, bio-assay-guided isolation led to the isolation and identification of five known compounds with a purity of more than 98%, and quercetin was both the best free radical scavenger and lipid-lowering constituent. Moreover, the mechanism of the lipid-lowering effect of AC30% might be its reduction in mRNA expression levels of sterol regulatory element binding protein 2 (SREBP2) and FAS gene in lipid synthesis. Otherwise, reducing the mRNA expression level of lipid synthesis genes, including SREBP1, SREBP2, fatty acid synthetase (FASN), β-Hydroxy β-methylglutaryl-CoA (HMGCR), stearoyl CoA desaturase 1 (SCD1), and increasing the mRNA expression level of lipid decomposition gene, such as carnitine palmitoyl transferease-1 (CPT1), might be involved in the lipid-lowering activity of quercetin. This study suggested that Allium cepa might be used to prevent and treat oxidative stress and dislipidemia-related disorders, including NAFLD.

Anti-inflammatory material basis and mechanism of Artemisia stolonifera based on UPLC-Q-TOF-MS combined with network pharmacology and molecular docking

This study aimed to explore the anti-inflammatory material basis and molecular mechanism of Artemisia stolonifera based on the analysis of the chemical components in different extracted fractions of A. stolonifera and their antioxidant and anti-inflammatory effects in combination with network pharmacology and molecular docking. Thirty-two chemical components were identified from A. stolonifera by ultra-performance liquid chromatography coupled to tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Among them, there were 7, 21 and 22 compounds in water, n-butanol and ethyl acetate fractions, respectively. The antio-xidant capacity of different extracted fractions was evaluated by measuring their scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) free radicals and total antioxidant capacity [ferric reducing antioxidant power(FRAP) assay]. The inflammatory model of RAW264.7 cells was induced by lipopolysaccharide(LPS), and the levels of nitrite oxide(NO), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6) in the supernatant and the mRNA expression of related inflammatory factors in cells were used to evaluate the anti-inflammatory effects. The results revealed that ethyl acetate fraction of A. stolonifera was the optimal antioxidant and anti-inflammatory fraction. By network pharmacology, it was found that flavonoids such as rhamnazin, eupatilin, jaceosidin, luteolin and nepetin could act on key targets such as TNF, serine/threonine protein kinase 1(AKT1), tumor protein p53(TP53), caspase-3(CASP3) and epidermal growth factor receptor(EGFR), and regulate the phosphatidylinositol-3-kinase-protein kinase B(PI3K-AKT) and mitogen-activated protein kinase(MAPK) signaling pathways to exert the anti-inflammatory effects. Molecular docking further indicated excellent binding properties between the above core components and core targets. This study preliminarily clarified the anti-inflammatory material basis and mechanism of ethyl acetate fraction of A. stolonifera, providing a basis for the follow-up clinical application of A. stolonifera and drug development.

Dialysis and column chromatography for biomass pyrolysis liquids separation

In the current study, a novel approach for separating value-added chemicals from pine wood residues’ pyrolysis liquids (bio-oil) was effectively carried out. It combined two separation techniques used for the first time in this field: dialysis with water, methanol and acetone, and column chromatography with Amberlite™ XAD7 resin. This strategy made it possible to separate bio-oil into four fractions: (1) pyrolytic lignin, which can be utilized in the synthesis of resins, foams, electrodes, asphalt, etc. (2) acid-rich fraction, with particular relevance to the chemical industry, (3) antioxidant fraction, containing phenolic compounds, with a lot of interest for pharmaceutical and nutraceutical industry, and (4) a final fraction containing the most non-polar chemicals from bio-oil. Thus, it was possible to develop a process that allows the obtention of bioproducts from woody biomass, a residue obtained in significant quantities in the management of non-profitable forests, making a step forward within the context of circular economy and bioeconomy.

Polysaccharides extracted with hot water from wild Prunus spinosa L. berries

Wild blackthorn berries represent an unexplored area in terms of the characterization of the natural biologically active polysaccharide complexes they contain. The antioxidant active fraction extracted from wild blackthorn fruits by hot water extraction (Hw) was subjected to ion-exchange chromatography and yielded six fractions by successive elution with salts. The purified fractions differed in the content of neutral sugars, uronic acids, proteins and phenolics. About 62% of the applied material was recovered from the column, with a higher yield of the fractions eluted with 0.25 M NaCl. Based on the sugar composition of the eluted fractions, several polysaccharide types were observed. The dominant components of Hw are the fractions eluted with 0.25 M NaCl (∼70%), which represent highly esterified homogalacturonan, containing up to 70–80% of galacturonic acids with a low content of rhamnogalacturonan associated with arabinan, galactan or arabinogalactan side chains, but no phenolics. Further, a dark brown polysaccharide material with a yield of ∼17% and with a high content of phenolic compounds, was eluted with alkali (1.0 M NaOH). It mainly represents an acidic arabinogalactan.

Revalorization of Colombian purple corn Zea mays L. by-products using two-step column chromatography

Colombian purple corn Zea mays L. by-products have been chemically characterized. To achieve this, after Amberlite XAD-7 purification and Sephadex LH-20 fractionation, the accurate anthocyanin and flavonol profile using UHPL-DAD-ESI-MS, total monomeric anthocyanin (TMA), polyphenols using Folin-Ciocalteau reduction capacity (FCRC), and antioxidant activity (DPPH and TEAC) of each fraction were performed. Cob and leaves illustrated a more complete flavonoid profile and a higher content of anthocyanins and flavonols, strongly related to the highest reducing power and radical scavenging activity compared to grains. Furthermore, the most antioxidant fractions corresponded to the higher molecular weight compounds. The cob and leaves were enriched in cyanidin-3-O-glucoside, cyanidin-3-O-malonyl-hexoside, and peonidin-3-O-glucoside, peonidin-3-O-(6′'-malonyl-glucoside). The purification and fractionation allowed us to establish the chemical and antioxidant characterization, and the resulting revalorization, of purple corn by-products for the first time, and to have available pure fractions of Zea mays L. for a wide diversity of industries.

Identification of Antioxidative Peptides Derived from Arthrospira maxima in the Biorefinery Process after Extraction of C-Phycocyanin and Lipids.

Arthrospira maxima has been identified as a sustainable source of rich proteins with diverse functionalities and bioactivities. After extracting C-phycocyanin (C-PC) and lipids in a biorefinery process, the spent biomass still contains a large proportion of proteins with potential for biopeptide production. In this study, the residue was digested using Papain, Alcalase, Trypsin, Protamex 1.6, and Alcalase 2.4 L at different time intervals. The resulting hydrolyzed product with the highest antioxidative activity, evaluated through their scavenging capability of hydroxyl radicals, superoxide anion, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS), was selected for further fractionation and purification to isolate and identify biopeptides. Alcalase 2.4 L was found to produce the highest antioxidative hydrolysate product after four-hour hydrolysis. Fractionating this bioactive product using ultrafiltration obtained two fractions with different molecular weights (MW) and antioxidative activity. The low-molecular-weight fraction (LMWF) with MW <3 kDa had higher DPPH scavenging activity with the IC50 value of 2.97 ± 0.33 compared to 3.76 ± 0.15 mg/mL of the high-molecular-weight fraction (HMWF) with MW >3 kDa. Two stronger antioxidative fractions (F-A and F-B) with the respective significant lower IC50 values of 0.83 ± 0.22 and 1.52 ± 0.29 mg/mL were isolated from the LMWF using gel filtration with a Sephadex G-25 column. Based on LC-MS/MS analysis of the F-A, 230 peptides derived from 108 A. maxima proteins were determined. Notably, different antioxidative peptides possessing various bioactivities, including antioxidation, were detected with high predicted scores together with in silico analyses on their stability and toxicity. This study established knowledge and technology to further value-add to the spent A. maxima biomass by optimizing hydrolysis and fraction processes to produce antioxidative peptides with Alcalase 2.4 L after two products already produced in a biorefinery. These bioactive peptides have potential applications in food and nutraceutical products.