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Multiple sclerosis (MS) is the leading cause of disability in young adults, with about 2.5-3 million cases currently diagnosed. Pain is a common comorbid symptom of MS, and develops independently from motor impairments. Currently utilized drugs bear severe side effects; thus, new therapeutic strategies are needed, and nutraceutical supplements represent innovative and safe opportunities. We have selected a variety of anthocyanin-enriched purple corn, from which a water-soluble extract (Red extract) has been obtained and administered to male rats exposed to experimental autoimmune encephalomyelitis (EAE). Animals developed relapsing-remitting motor symptoms, accompanied by early onset trigeminal allodynia. The preventive administration of Red extract facilitated the remission of motor symptoms, prevented the development of relapses, and delayed and reduced the development of EAE-associated trigeminal pain. An overall inhibition of neuroinflammation, blunted microgliosis and astrogliosis, activation of autophagy and reduced immune cell infiltration in the brainstem, cervical and lumbar spinal cord were observed. Yellow corn extract, lacking anthocyanins, had no behavioral effects, despite a limited anti-inflammatory action. Therapeutic Red extract administration did not affect EAE motor symptoms, only partially reduced the development of trigeminal pain but maintained its ability to reduce neuroinflammation and glial cell activation and to promote autophagy. Overall, our data suggest that a nutraceutical supplement from anthocyanin-enriched purple corn represents an interesting option to limit the development of motor relapses and the chronicization of multiple sclerosis-associated pain, through the mitigation of neuroinflammation, of the infiltration of immune cells and the promotion of autophagy., Competing Interests: Declaration of Competing Interest None., (Copyright © 2025 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

In this work, new sorbents for the purification of anthocyanin-rich extracts were evaluated. Copolymers of 4-vinylpyridine crosslinked with trimethylolpropane trimethacrylate (poly(4VP-co-TRIM)) or 1,4-dimethacryloyloxybenzene (poly(4VP-co-14DMB)) were tested for their potential to capture polyphenols. Copolymers were obtained by seed swelling polymerization in the form of microspheres with permanent porous structure - attractive features of sorbents used for sample purification by dispersive solid phase extraction. The microspheres were characterized by AFM, elemental analysis, SEM, and nitrogen adsorption-desorption method. Their capacity to remove polyphenols was evaluated using spectrophotometry, HPLC-DAD, and LC-MS/MS. For proof-of-concept, the aqueous extracts of berries classified into three different groups regarding their anthocyanin composition (strawberries, raspberries, blackcurrants) were selected. It was found that studied microspheres adsorbed flavonoids more effectively compared to primary secondary amine and graphitized carbon black. Copolymers of 4-vinylpyridine also capture anthocyanins and might be used for the purification of extracts of fruits before LC-MS/MS analysis to reduce the matrix effect., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ilona Sadok, Agnieszka Krzyszczak-Turczyn, Marta Grochowicz have patent “Method of obtaining purified berry fruit extracts and the use of microspheres based on 4-vinylpyridine for the purification of berry fruit extracts”, P.448492 pending to The Patent Office of the Republic of Poland., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Pistacia lentiscus L., commonly known as the mastic tree or lentisk, is a woody Mediterranean plant revered for its ecological relevance as well as for its extensive ethnobotanical heritage. Historically, the fruits and the resin of P. lentiscus have been widely utilized in traditional medicine, underscoring its important role in local healing practices. Given these properties, this study explored an innovative approach to efficiently extract anthocyanins and flavonols from P. lentiscus oilcakes utilizing ultrasound-assisted extraction (UAE) as an alternative to conventional solvent extraction. Liquid chromatography-mass spectrometry (LC-MS) and high-performance liquid chromatography with diode-array detection (HPLC-DAD) were used to identify and quantify the anthocyanins and flavonols, revealing the successful extraction of eight distinct anthocyanins and twenty flavonols. A Fractional Factorial Design (FFD) followed by a Box-Behnken design (BBD) were applied to optimize the yield of anthocyanins and flavonols. The optimal extraction conditions found were to be an extraction time of 15 min with 70% ethanol as the solvent and a liquid-to-solid ratio of 0.012 L g -1 , which resulted in a maximum extraction yield of 19.78 mg g -1 dry extract for the Total Flavonol Content and over 25.4 mg g -1 dry extract for the Total Flavonol and Anthocyanin Content. By elucidating the optimal conditions for extracting anthocyanins and flavonol glycosides, this study opens promising avenues for utilizing P. lentiscus oilcake by-products, supporting sustainable practices, and advancing the valorization of Mediterranean bio-resources for health-promoting applications.

Maejo 341 Sweet potato (MSP) is a new purple sweet potato variety cultivated in Northern Thailand, but its health benefits are unknown. This study aimed to investigate its antioxidant, anti-inflammatory, and anti-osteoporotic activities, as well as its anthocyanin content. The peel and flesh of MSP were extracted with ethanol and water. Compared with the flesh extracts, the peel extracts presented greater antioxidant capacity and were rich in phenolics, flavonoids, and anthocyanins, namely, cyanidin-3-O-glucoside, peonidin-3-O-glucoside, pelargonidin-3-O-glucoside, cyanidin, and peonidin. The peel extracts suppressed lipopolysaccharide-induced inflammation by inhibiting the secretion of proinflammatory cytokines and enzymes, including TNF-α, IL-1β, IL-6, COX-2, and iNOS, as well as reducing nitric oxide and matrix metalloproteinase-9 secretion. The extracts inhibited the RANKL-induced NF-κB and MAPK pathways and downregulated osteoclastogenic marker expression. Under LPS and RANKL treatment, the peel extracts notably reduced reactive oxygen species production while increasing antioxidant gene expression. Furthermore, they increased osteoblast viability and slightly raise alkaline phosphatase activity. These findings suggest that MSP peel could be used as a functional food to alleviate oxidative stress and inflammation-related osteoporosis., Competing Interests: Declarations. Conflict of interest: The authors declare that there are no conflicts of interest., (© 2025. The Author(s).)

Black rice bran (BRB), a valuable byproduct from the rice milling process, possesses numerous pharmacological activities, including antioxidant potential, but information regarding highly efficient extraction methods is scarce. To enhance the extraction efficiency, ultrasonic-assisted extraction coupled with Box-Behnken design (BBD) was used in this study to maximize the total phenolic content (TPC), total flavonoid content (TFC), total anthocyanin content (TAC), and antioxidant capacity of BRB extract. The BBD results showed that 57% ethanol at 50°C and pH 3.1 were best for TPC (136.2 mg GAE/g dry weight), TAC (2.3 mg cyanidin-3-O-glucoside equivalents/g dry weight), and TFC (38.6 mg quercetin equivalents/g dry weight). In vitro antioxidant capacities of BRB extract determined by 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS + ) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays revealed free radical scavenging potentials with EC 50 values of 67.62 and 1149 µg/mL, respectively. The cytotoxic effect of BRB extract on human skin HaCaT keratinocytes was determined by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, which revealed no significant cytotoxicity and a significant increase in cell viability at a concentration of 500 µg/mL. Ultra-high performance liquid chromatography (UHPLC) analysis identified syringic acid, gallic acid, protocatechuic acid, vanillic acid, p-coumaric acid, and ferulic acid in BRB extract. This study provides the optimal conditions to extract antioxidant bioactive compounds from BRB and supports the use of BRB as a source of antioxidants. Due to BRB showing no significant cytotoxicity to human skin cells, it could potentially be further developed for topical applications., (© 2024 Institute of Food Technologists.)

This study determined the anthocyanin and phenolic profile of Syzygium cumini bioactive compounds, including anthocyanins and other flavonoids, alongside diverse phenolic compounds. The study optimized a green extraction technique (ultrasound-assisted enzymatic extraction (UAEE)) to obtain anthocyanin-rich extract from the fruit pulp of S. cumini using the pectinase enzyme. UHPLC-LC/MS, FTIR, and SEM were used to profile the secondary metabolites, functional groups, and surface morphology. Two major anthocyanins, cyanidin and malvidin, and twenty-three non-anthocyanins, including gallic acid, naringenin, myricetin, and kaempferol, were identified in the enzymatic extract of S. cumini. A central-composite design was used to optimize the extraction, analyzing the effects of enzyme concentration (0.01-0.03 %), pH (1-3), and ultrasonication time (5-15 min) on total anthocyanin content (438.75 ± 29.81 mg C3G/100 g db), determining the optimal points (0.01 %, 2 pH and 10 mins). The optimized extract was further investigated for total phenolic content and antioxidant activities. The study utilized an economical approach to effectively extract maximum anthocyanins from S. cumini fruit for their potential applications as a biocolorant in food products, simultaneously establishing promising health potential through available literature., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

3-Deoxyanthocyanidins (3-DAS) are the unique flavonoid compounds in sorghum. The majority of research to date have concentrated on the biological activity and extraction of 3-DAS from sorghum and lacked systematic purification and identification investigations. Herein, a facile method for the purification of sorghum 3-DAS from the acidic methanol solution crude extract using macroporous resins (MARs) was proposed and investigated in this work. Based on the experimental data, commercial MAR AB-8 was selected to achieve the purification of sorghum 3-DAS. The adsorption-desorption experiment confirmed the optimal procedure. With this procedure, the total purity of 3-DAS was 19.32%, and 3-DAS content in the purified product was 8.4 times higher than that in the crude extract. UPLC-QTRAP-MS/MS identification indicated that the purified 3-DAS product was mainly composed of apigeninidin, luteolinidin, and their methoxy derivatives. Besides, the purified product exhibited better bioactivities than the crude extract, including the scavenging activities for DPPH, ABTS + , and hydroxyl radicals, the total antioxidant capacity, and the inhibitory effects for α-amylase, α-glucosidase, and lipase. This work provides a potential way for the large-scale purification of bioactive sorghum 3-DAS from crude extract, which is significant to deepen the applications of sorghum 3-DAS in the food nutrition and pharmaceutical industries., (© 2024 Institute of Food Technologists.)

Background: Blackberry seeds, as a by-product of processing, have potential bioactive substances and activities. A response surface method was used to determine the optimal conditions of blackberry seed extracts (BSEs) with high 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity by ultrasound-assisted extraction (UAE). The composition and antioxidant capacity of BSEs were further analyzed., Results: The optimal conditions were material-to-liquid ratio of 0.07 g mL -1 , ethanol concentration of 56%, extraction temperature of 39 °C and ultrasonic power of 260 W. Using these conditions, the extraction yield and total polysaccharide, phenolic and anthocyanin contents in BSEs were 0.062 g g -1 and 633.91, 36.21 and 3.07 mg g -1 , respectively. The Fourier transform infrared spectra of BSEs exhibited characteristic peaks associated with polysaccharide absorption. The antioxidant capacity, DPPH and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, and ferric reducing antioxidant power of BSEs were 1533.19, 1021.93 and 1093.38 mmol Trolox equivalent g -1 , respectively. The delphinidin-3-O-glucoside, paeoniflorin-3-O-glucoside and cyanidin-3-O-arabinoside contents in BSEs were 3.05,12.76 and 1895.90 ± 3.45 μg g -1 . Five polyphenols including gallic acid, coumaric acid, ferulic acid, catechin and caffeic acid were identified and quantified in BSEs with its contents at 8850.43, 5053.26, 4984.65, 1846.91 and 192.40 μg g -1 ., Conclusion: These results provide a method for preparing BSE containing functional components such as polysaccharides, phenols and anthocyanins through UAE, and BSEs have potential application in food industries. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The aim of this study is to develop roselle (Hibiscus sabdariffa L.) extracts with high amounts of phytochemicals and in vitro antioxidant and antibacterial effects using green extraction technique. Roselle extract prepared by ultrasonic extraction with 80% ethanol and 1% hydrochloric acid (RSUEH) promoted the highest yield and high total phenolic (5.21 ± 0.05 g% gallic acid equivalent of dried extract), total flavonoid (1.29 ± 0.06 g% quercetin equivalent of dried extract), and total anthocyanin contents (0.21 ± 0.00 and 0.44 ± 0.00 g% cyanidin 3-glucoside equivalent of dried extract determined by pH-differential method with standard calculation and pH-differential method with standard curve of cyanidin 3-glucoside, respectively). From the validated high-performance liquid chromatography analysis, delphinidin-3-sambubioside, delphinidin-3-glucoside, and cyanidin-3-glucoside were found to be the major anthocyanins in roselle extract. This extract exhibited moderate 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging effect (EC 50 289.61 ± 0.16 µg/mL) and strong inhibitory effects against Escherichia coli, Staphylococcus aureus, Staphylococcus intermedius, Proteus mirabilis, and Pseudomonas aeruginosa with the inhibitory concentration (IC 50 ) values in the range of 0.15 ± 0.05 to 4.64 ± 0.07 mg/mL and the minimum inhibitory concentration (MIC) values ranged from 1.66 ± 0.06 to 7.11 ± 1.05 mg/mL. From partial purification, fraction 28 (F28) contained higher amounts of all analyzed phytochemicals than roselle extracts and promoted stronger DPPH scavenging effects (EC 50 54.88 ± 2.23 µg/mL). F28 showed antibacterial activity with IC 50 values against all tested bacteria in the range of 0.19 ± 0.07 to 4.69 ± 0.09 mg/mL, and the MIC values ranged from 2.46 ± 0.08 to 6.24 ± 0.09 mg/mL., (© 2024 The Author(s). Journal of Food Science published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

In addition to their pigment properties, the potential health benefits of anthocyanins have made them a subject of interest in recent years. This study aimed to obtain purified anthocyanin fractions from native Mexican black bean cultivars using Amberlite XAD-7 resin column and HPCCC and evaluate their anti-inflammatory properties using RAW 264.7 cells. The major anthocyanins in the purified anthocyanin fractions were delphinidin 3-glucoside (61.8%), petunidin 3-glucoside (25.2%), and malvidin 3-glucoside (12.2%). Purified anthocyanin fractions at 12.5 μg/mL effectively prevented LPS-induced ERK1/ERK2 phosphorylation and reduced the protein expression of COX-2 and mRNA expression of iNOS. Results showed that purified anthocyanin fractions have the potential to modulate the inflammatory response by inhibiting the production of pro-inflammatory mediators through the ERK1/ERK2 and NF-κB pathways. This study suggests that anthocyanins from black beans could be used as a natural strategy to help modulate inflammation-associated diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Butterfly pea flower (BPF), roselle calyx (RC), and grape skin (GS) are rich in bioactive phenolics with health benefits. Due to its simplicity, safety, and environmental friendliness, this study used water as a solvent to explore different extraction conditions in these plant materials and compared the heat stability of anthocyanins in the aqueous extracts. To maximize the total anthocyanins and polyphenols in the aqueous extracts, the powders of BPF, GS, and RC should be extracted for 30 min at 90 °C; 30 min and 120 min at 90 °C; and 30 min and 60 min at 60 °C, respectively. Among the tested plant materials, the content of total anthocyanins was RC > GS > BPF, while the total phenolic content was GS > BPF > RC. Anthocyanins of the aqueous extracts underwent rapid thermal degradation at high temperatures and high pH values. The thermal stability of anthocyanins in the materials was in the order: BPF > GS > RC. This is likely related to the types and structures of the anthocyanins such as the degree of acylation and glycosylation. The study demonstrates that hot water extraction is efficient and practical for these materials, yielding extracts suitable for food and nutraceutical applications.

Background: To extract anthocyanins with high efficiency, a hypothesis for high-speed shear homogenization extraction (HSHE) method was established through a combination of solvent and ultrasonic-assisted extractions. The efficacy of this hypothesis was demonstrated by performing qualitative and quantitative analyses of 16 anthocyanins extracted from five northern vegetables, and five berry fruits using ultra-high-performance Q-Exactive Orbitrap tandem mass spectrometry. Single-factor experiments were conducted by varying ethanol concentration, temperature, pH and extraction cycles to determine the optimal conditions for this method., Results: Optimal extraction conditions (ethanol 70-80%, 40-50 °C, pH 3-4, performed twice) were determined using an HSHE (5 min, 10 000 rpm, 25 °C) assisted shaker (60 min) and ultrasonication (40 kHz, 160 W cm -2 , 30 min, 25 °C) procedure. Compared to the traditional non-HSHE method, the total anthocyanin content obtained through HSHE extraction showed a significant increase, ranging from 1.0 to 3.9 times higher, with purple cabbage exhibiting the most pronounced enhancement in content. More types of anthocyanins were detected in blueberry (9), black bean (7) and raspberry (5), of which malvidin was the major anthocyanin (0.426 g kg -1 ) in blueberry, having an amount five times than previously obtained., Conclusion: The established HSHE method has been proven to be a superior technique for anthocyanin extraction, with higher extraction efficiency and concentrations. This technique also provides a new avenue for extracting bioactive compounds from diverse food sources, with potential applications in improving the functional properties of food products. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Cyanidin-3-O-galactoside (Cy3-gal) is the most widespread anthocyanin that has been found to be applicable to nutraceutical and pharmaceutical ingredients. Nevertheless, the process of separation and purification, susceptibilities to heat, and pH inactivation present some limitations. In the present study, natural deep eutectic solvents (NADES) with an ultrasonic-assisted extraction method were briefly studied, and the recovery of Cy3-gal from Rhododendron arboreum was highlighted. The NADES, consisting of choline chloride and oxalic acid (1:1), was screened out as an extractant, and single-factor experiments combined with a two-site kinetic model were employed to describe the extraction process. Further, the work investigated ultrasound-assisted adsorption/desorption to efficiently purify Cy3-gal using macroporous resins. The optimal extraction conditions to attain maximum Cy3-gal yield was 30% water in a 50:1 (mL/g) solvent-to-sample ratio, 11.25 W/cm 3 acoustic density, and 50% duty cycle for 16 min of extraction time. Under these conditions, the results revealed 23.07 ± 0.14 mg/g of Cy3-gal, two-fold higher than the traditional solvents. Furthermore, of the different resins used, Amberlite XAD-7HP showed significantly (p < 0.05) higher adsorption/desorption capacities (12.82 ± 0.18 mg/g and 10.97 ± 0.173 mg/g) and recovery (48.41 ± 0.76%) percent over other adsorbents. Experiments on the degrading behavior (40-80 °C) of the recovered Cy3-gal were performed over time, and the first-order kinetic model better explained the obtained data. In conclusion, the study asserts the use of ultrasonication with NADES and XAD-7HP resin for the improved purification of Cy3-gal from the crude extract., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In the context of a circular economy, this study explores the valorization of blueberry pomace (BP) as a source of bioactive compounds using sustainable extraction methods. Microwave-assisted extraction (MAE) and microwave-assisted subcritical water extraction (MASWE) were employed to obtain two distinct fractions: MAE 1° and MASWE 2°. The first extract, MAE 1°, obtained at 80 °C, had a high total anthocyanin content (21.96 mg Cya-3-glu /g extract ), making it suitable as a natural pigment. Additionally, MAE 1° exhibited significant enzyme inhibition, particularly against α-amylase and β-glucosidase, suggesting potential anti-diabetic and anti-viral applications. The second extract, MASWE 2°, obtained at 150 °C, contained a higher total phenolic content (211.73 mg GAE /g extract ) and demonstrated stronger antioxidant activity. MASWE 2° showed greater inhibition of acetylcholinesterase and tyrosinase, indicating its potential for use in Alzheimer's treatment, skincare, or as a food preservative. MASWE 2° exhibited cytotoxicity against HeLa cells and effectively mitigated H 2 O 2 -induced oxidative stress in HaCat cells, with MAE 1° showing similar but less pronounced effects. A tested formulation combining MAE 1° and MASWE 2° extracts in a 3:2 ratio effectively enhanced anthocyanin stability, demonstrating its potential as a heat-stable pigment. The extract characteristics were compared with a conventional method (MeOH-HCl in reflux condition), and the protocol's sustainability was assessed using several green metric tools, which provided insights into its environmental impact and efficiency.

Jabuticaba peel, rich in antioxidants, offering health benefits. In this study, the extraction of phenolic compounds from jabuticaba peel using ultrasound-assisted (UA) and their subsequent concentration by nanofiltration (NF) employing a polyamide 200 Da membrane was evaluated. The UA extractions were conducted using the Central Composite Rotatable Design (CCRD) 22 methodology, with independent variables extraction time (11.55 to 138 min) and temperature (16.87 to 53.3 °C), and fixed variables mass to ethanol solution concentration at pH 1.0 (1:25 g/mL), granulometry (1 mm), and ultrasonic power (52.8 W). The maximum concentrations obtained were 700.94 mg CE/100 g for anthocyanins, 945.21 mg QE/100 g for flavonoids, 133.19 mg GAE/g for phenols, and an antioxidant activity IC50 of 24.36 μg/mL. Key phenolic compounds identified included cyanidin-3-glucoside, delphinidin-3-glucoside, and various acids like syringic and gallic. NF successfully concentrated these compounds, enhancing their yield by up to 45%. UA and NF integrate for sustainable extraction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Recently, deep eutectic solvents (DES) have been extensively researched as a more biocompatible and efficient alternative to conventional solvents for extracting pigments from natural resources. The efficiency of DES extraction for the anthocyanin and carotenoid can be enhanced by microwave-assisted extraction (MAE) and/or ultrasound-assisted extraction (UAE) techniques. Apart from the extraction efficiency, the toxicity and recovery of the pigments and their bioavailability are crucial for potential applications. A plethora of studies have explored the extraction efficiency, toxicity, and recovery of pigments from various natural plant-based matrices using DES. Nevertheless, a detailed review of the deep eutectic solvent extraction of natural pigments has not been reported to date. Additionally, the toxicity, safety, and bioavailability of the extracted pigments, and their potential applications are not thoroughly documented. Therefore, this review is designed to understand the aforementioned concepts in using DES for anthocyanin and carotenoid extraction., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. For the purpose of open access, the author, Ali Ali Redha, has applied a ‘Creative Commons Attribution (CC BY) licence to any Author Accepted Manuscript version arising., (Crown Copyright © 2024. Published by Elsevier Ltd. All rights reserved.)

A dynamic compressed fluid-based separation process combining carbon dioxide and ethanol was explored to isolate portisins previously hemi-synthesized from blueberry surplus anthocyanins. The influence of process parameters such as pressure (100-500 bar), temperature (40-60 °C), and ethanol content in the compressed fluid mixture (20-50 wt%) on extraction yield, portisins yield, and portisins content in the extract was investigated. The two-step isolation process includes (1) a first step at 100 bar, 60 °C, and 20 wt% ethanol content in the compressed fluid mixture to remove the low polarity compounds; and (2) a second step at 500 bar, 40 °C, and 100 wt% ethanol to recover portisins, resulting in a 1.5-fold increase in portisins content. The performance of the two-step separation process was compared to centrifugal partitional chromatography and conventional reverse phase liquid chromatography already reported in terms of portisins content in the extract, process throughput, process efficiency, and total solvent used. The two-step separation process decreased the total solvent used, although with a decrease in the throughput and efficiency. Nevertheless, the choice of the best separation technology depends on the application, as these techniques result in different portisins purities. Overall, this study contributed to a scalable and more sustainable process for natural colorant production, specifically focusing on blue pigments, with several industrial applications.

Saffron ( Crocus sativus ) floral by-products are a source of phenolic compounds that can be recovered and used in the nutraceutical, pharmaceutical, or cosmetic industries. This study aimed to evaluate the phenolic compounds' extraction using green extraction techniques (GETs) in saffron floral by-products and to explore the influence of selected extraction techniques on the phytochemical composition of the extracts. Specifically, ultrasound-assisted extraction (UAE), subcritical water extraction (SWE), and deep eutectic solvents extraction (DESE) were used. Phenolic compounds were identified with (HR) LC-ESI-QTOF MS/MS analysis, and the quantitative analysis was performed with HPLC-PDA. Concerning the extraction techniques, UAE showed the highest amount for both anthocyanins and flavonoids with 50:50% v / v ethanol/water as solvent (93.43 ± 4.67 mg/g of dry plant, dp). Among SWE, extraction with 96% ethanol and t = 125 °C gave the best quantitative results. The 16 different solvent mixtures used for the DESE showed the highest amount of flavonoids (110.95 ± 5.55-73.25 ± 3.66 mg/g dp), while anthocyanins were better extracted with choline chloride:butane-1,4-diol (16.0 ± 0.80 mg/g dp). Consequently, GETs can be employed to extract the bioactive compounds from saffron floral by-products, implementing recycling and reduction of waste and fitting into the broader circular economy discussion.

In this paper, biological investigations and a high-resolution UPLC-PDA-ESI-qTOF-HRMS technique were employed for Brassica oleracea L. var. capitata f. rubra DC. (red cabbage) of the family Brassicaceae (Cruciferae), cultivated in Egypt, for the first time. The positive ionization mode is usually performed to identify anthocyanins. However, this technique cannot differentiate between anthocyanins and corresponding non-anthocyanin polyphenols. Thus, the negative ionization mode was also used, as it provided a series of characteristic ions for the MS analysis of anthocyanins. This helped in identifying five kaempferol derivatives for the first time in red cabbage, as well as nine-previously reported-anthocyanins. For the biological investigations, the acidified methanolic extract of fresh leaves and the methanolic extract of air-dried powdered leaves were examined for their antioxidant, antimicrobial, and anticancer activities. The freshly prepared phenolic extract was proven to be more biologically potent. Statistical significance was determined for its anticancer activity in comparison with standard doxorubicin.

Age-related macular degeneration (AMD) is one of the major causes of blindness in elderly populations. However, the dry form of AMD has lack of effective treatments. The fruits of Aronia melanocarpa are rich in anthocyanins. In this study, the protective effects of aronia fruit extract on rat retina were investigated using a NaIO 3 -induced dry AMD model. Full-field electroretinograms (ERGs) showed that b-wave amplitudes were significantly decreased and the retina structures were disordered in the model. The extract treatment alleviated the injuries. The b-wave amplitudes increased 61.5% in Scotopic 0.01ERG, 122.0% in Photopic 3.0ERG, and 106.8% in Photopic 3.0 flicker; the retina structure disorder was improved with the thickness of outer nuclear layer increasing by 44.1%; and the malonaldehyde level was significantly reduced in extract-treated rat retinas compared to the model. The proteomics analysis showed the expressions of five crystallin proteins, α-crystallin A chain, β-crystallin B2, β-crystallin A3, α-crystallin B chain, and γ-crystallin S, which protect retina ganglion cells, were increased by 7.38-, 7.74-, 15.30-, 4.86-, and 9.14-fold, respectively, in the extract treatment compared to the control, which was also confirmed by immunoblotting. The results suggest that aronia fruit extract, probably due to its anthocyanins, could protect the rat retina by alleviating oxidative damages and by upregulating the crystallin proteins to protect its nerve system.

Anthocyanins from flowers of the butterfly pea ( Clitoria ternatea L.) are promising edible blue food colorants. Food processing often faces extreme pHs and temperatures, which greatly affects the color and nutritional values of anthocyanins. This study explored the color, spectra, storage stability, and antioxidant properties of C. ternatea anthocyanin extract (CTAE) at different pHs. The color and absorption spectra of CTAEs at a pH of 0.5-13 were shown, with their underlying structures analyzed. Then, the storage stability of CTAEs were explored under a combination of pHs and temperatures. The stability of CTAE declines with the increase in temperature, and it can be stored stably for months at 4 °C. CTAEs also bear much resistance to acidic and alkaline conditions but exhibit higher thermal stability at pH 7 (blue) than at pH 0.5 (magenta) or pH 10 (blue-green), which is a great advantage in food making. Antioxidant abilities for flower extracts from the butterfly pea were high at pH 4-7, as assessed by DPPH free radical scavenging assays, and decreased sharply when the pH value exceeded 7. The above results provide a theoretical basis for the application of butterfly pea flowers and imply their great prospect in the food industry.

To extract anthocyanins from wild Prunus cerasifera fruit peel and to identify the main compositions. The effects of extraction solvent, solid-to-liquid ratio, extraction time and extraction temperature on the extraction rate of anthocyanins were determined using colorimetric method, differential spectrophotometry and L9 (34) orthogonal experiment. The structure of the anthocyanins extracted from wild Prunus cerasifera fruit peel was preliminarily identified by liquid chromatography-electrospray ionization-tandem mass spectrometry. And hydroxyl radical scavenging rate and DPPH radical scavenging rate were used to investigate the antioxidant activity of Xinjiang cherry anthocyanin in vitro. The highest extraction rate of anthocyanins was acquired under the conditions including 1% HCl-methanol, solid-to-liquid ratio =1:5, extraction temperature 55°C and extraction time 80 minutes. Liquid chromatography-electrospray ionization-tandem mass spectrometry results showed that the main compositions of anthocyanins were cyanidin 3-galactoside, cyanidin 3-glucoside, cyanidin 3-rutinoside and cyanidin 3-xyloside. The results of antioxidant test showed that Xinjiang cherry anthocyanin had certain scavenging effect on 1, 1-diphenyl-2-picrylphenylhydrazine (DPPH•) and hydroxyl radical (•OH), but was weaker than Vc. Findings from this study provide evidence for extraction process and composition identification of anthocyanins from Xinjiang wild Prunus cerasifera fruit peel.

Mulberry (Morus alba L.) fruit (MF) is a rich source of functional compounds, such as anthocyanin. However, during solvent extraction, these compounds are not fully dispersed into the substrate, leading to incomplete extraction. Moreover, raw MF rapidly ripens and deteriorates after harvesting; hence, innovative methods to process MF are needed. Here, a pectinase-assisted extraction method is developed to liberate polyphenols and anthocyanins from cell wall matrices in MF. We optimized the procedure to maximize water solubility index (WSI), total phenolic (TP) content, and total anthocyanin (TA) content using a central composite design to perform a response surface methodology (RSM) analysis. The optimal conditions predicted by the RSM were a 1:5 w/v material/water ratio with 3.5% pectinase (v/w) and 1.5% citric acid (w/w) for 113 min at 50°C. Under these conditions, the WSI, TP, and TA were significantly higher compared with those in the untreated control. The results well matched (within 5% differences) with the predicted RSM values. Furthermore, metabolite analysis revealed that the levels of cyanidin-3-O-glucoside, delphinidin hexoside, and quercetin were higher in pectinase-assisted MF extraction compared with the untreated control. This work demonstrated that pectinase-assisted extraction using citric acid could be an efficient technique to enhance the value of MF and its potential applications in the food industry. PRACTICAL APPLICATION: A pectinase-assisted extraction method was optimized to enhance the WSI, TP, and TA yields from MF extracts. The optimal conditions were predicted to be 1:5 w/v material/water ratio, 3.5% pectinase (v/w), and 1.5% CA (w/w) with a 113 min reaction time at 50°C. Under these conditions, WSI, TP, and TA were significantly increased compared with the untreated control. These results suggested the potential of mulberry plants for use in the food industry via the development of a simple, efficient process to extract functional compounds from MF., (© 2021 Institute of Food Technologists®.)

Phenolic compounds from plant sources have significant health-promoting properties and are known to be an integral part of folk and herbal medicines. Consumption of phenolics is known to alleviate the risk of various lifestyle diseases including cancer, cardiovascular, diabetes, and Alzheimer's. In this context, numerous plant crops have been explored and characterized based on phenolic compounds for their use as supplements, nutraceutical, and pharmaceuticals. The present review highlights some important source of bioactive phenolic compounds and novel technologies for their efficient extraction. These techniques include the use of microwave, ultrasound, and supercritical methods. Besides, the review will also highlight the use of response surface methodology (RSM) as a statistical tool for optimizing the recoveries of the phenolic bioactives from plant-based matrices., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Passion fruit rind is a waste product from the beverage industry that is rich in anthocyanins that can be potentially applied as a natural colorant. However, the inherent instability of anthocyanins causes rapid discoloration. In this study, the cyanidin-3-glucoside (C-3-G) in passion fruit rind was extracted using 50% ethanol and converted into nonbleachable pigments by reaction with Oolong tea extracts and acetaldehyde. Reactions over 70 days formed high concentrations of stable nonbleachable pigments (3.07-6.68 absorbance unit [A.U.], in total) such as pyranoanthocyanins, as well as oligomeric and polymeric pigments with ethyl-linked bridges. In C-3-G and acetaldehyde reaction, positive relations were found among acetaldehyde concentration, color density, and nonbleachable pigment concentrations. As for reactions with C-3-G and Oolong tea extract combined with acetaldehyde, greater color density and greater concentrations of nonbleachable pigments (10.80-12.34, 4.25-4.40 A.U., respectively) were formed compared with acetaldehyde alone. In addition, the antioxidant capabilities of the extracts were enhanced after reaction with Oolong tea extracts. The results of this study show a useful method to enhance the stability of anthocyanins from passion fruit rind and also provide greater economic value to this waste product. PRACTICAL APPLICATION: Ripened passion fruits contain a high concentration of anthocyanins in their rind. These anthocyanins can be optimally extracted by ultrasonic assisted solvent extraction to provide stable pigments by inducing acetaldehyde (a volatile compound often found in foods and beverages) into the anthocyanins. These stable pigments have a greater reddish hue in solution than the anthocyanin extracted from the rind and are more stable over a greater pH range. In addition, these stable pigments can be potentially used as colorant throughout the food and cosmetic industry to provide high economical values., (© 2021 Institute of Food Technologists®.)

Two common extraction solvent systems, namely acidified aqueous methanol and acidified aqueous acetone, were used to extract blackberry phenolics, and the antioxidant properties of the recovered extracts were compared. The crude extracts were fractionated into low- and high-molecular-weight phenolics by Sephadex LH-20 column chromatography. The hydrophilic-oxygen radical absorbance capacity (H-ORAC FL ), ferric reducing antioxidant power (FRAP), and the cellular antioxidant activity (CAA) assays were employed as indices to assess antioxidant capacity of the extracts and their respective fractions. The methanolic solvent system displayed a greater efficiency at extracting anthocyanin and flavonol constituents from the blackberries, while the acetonic solvent system was better at extracting flavan-3-ols and tannins. Anthocyanins were the dominant phenolic class found in the blackberries with 138.7 ± 9.8 mg C3G eq./100 g f.w. when using methanol as the extractant and 114.6 ± 3.4 mg C3G eq./100 g f.w. when using acetone. In terms of overall antioxidant capacity of blackberry phenolics, the acetonic solvent system was superior. Though present only as a small percentage of the total phenolics in each crude extract, the flavan-3-ols (42.37 ± 2.44 and 51.44 ± 3.15 mg/100 g f.w. in MLF and ALF, respectively) and ellagitannins (5.15 ± 0.78 and 9.31 ± 0.63 mg/100 g f.w. in MHF and AHF, respectively) appear to account for the differences in the observed antioxidant activity between the two solvent systems.

Background: Skin cell walls modulate anthocyanin and tannin extraction from grape skins. However, relationships between the composition of alcohol-insoluble cell wall solids (AIS) and extraction are still unclear. Our objectives were to characterize the impact of variety, berry size and ripeness on skin AIS composition (polysaccharides, proteins) and polyphenol extraction during maceration., Results: Grape skin composition and its impact on polyphenol extraction was compared for two varieties - Carignan and Grenache - with skins of berries sorted according to their size and density. Extractions were performed under model wine-like maceration conditions. Fresh skins had similar content of polymeric tannins, but strongly differed in their anthocyanin content (higher in Carignan and in the ripest berries) and composition (higher proportions in coumaroylated anthocyanins in Carignan). Anthocyanin extraction was proportionally much higher in Grenache, which was not just related to the Carignan's higher levels in coumaroylated anthocyanins. Chemical reactions decreased anthocyanin concentrations in solution for both varieties. Tannin extraction for Grenache was slightly higher and faster than for Carignan. Skin AISs differed slightly between the two varieties in their carbohydrate composition and protein content, but not between modalities. Polyphenol analyses in the precipitates evidenced at the end of the maceration and in residual skins highlighted differences between the two varieties and between berries with different ripeness., Conclusion: Structural information on the cell wall network and on its changes during maceration, along with a better understanding of the chemical reactions of anthocyanins and tannins, is needed to better relate grape and wine polyphenol composition. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Wastewaters and by-products generated in the winemaking process are important and inexpensive sources of value-added compounds that can be potentially reused for the development of new products of commercial interest (i.e., functional foods). This research was undertaken in order to evaluate the potential of nanofiltration (NF) membranes in the recovery of anthocyanins and monosaccharides from a clarified Carménère grape marc obtained through a combination of ultrasound-assisted extraction and microfiltration. Three different flat-sheet nanofiltration (NF) membranes, covering the range of molecular weight cut-off (MWCO) from 150 to 800 Da, were evaluated for their productivity as well as for their rejection towards anthocyanins (malvidin-3- O -glucoside, malvidin 3-(acetyl)-glucoside, and malvidin 3-(coumaroyl)-glucoside) and sugars (glucose and fructose) in selected operating conditions. The selected membranes showed differences in their performance in terms of permeate flux and rejection of target compounds. The NFX membrane, with the lowest MWCO (150-300 Da), showed a lower flux decay in comparison to the other investigated membranes. All the membranes showed rejection higher than 99.42% for the quantified anthocyanins. Regarding sugars rejection, the NFX membrane showed the highest rejection for glucose and fructose (100 and 92.60%, respectively), whereas the NFW membrane (MWCO 300-500 Da) was the one with the lowest rejection for these compounds (80.57 and 71.62%, respectively). As a general trend, the tested membranes did not show a preferential rejection of anthocyanins over sugars. Therefore, all tested membranes were suitable for concentration purposes.

Many current food and health trends demand the use of more ecological, sustainable, and environmentally friendly techniques for the extraction of bioactive compounds, including antioxidants. However, extraction yields and final antioxidant activities vary between sources and are highly influenced by the given extraction method and nature and ratio of the employed solvent, especially for total polyphenols, flavonoids, and anthocyanins, which are well recognized as natural antioxidants with food applications. This review focused on the most common extraction techniques and potential antioxidant activity in the food industry for various natural antioxidant sources, such as green tea, rosemary, clove, and oregano. Green extraction techniques have been proven to be far more efficient, environmentally friendly, and economical. In general, these techniques include the use of microwaves, ultrasound, high hydrostatic pressure, pulsed electric fields, enzymes, and deep eutectic solvents, among others. These extraction methods are described here, including their advantages, disadvantages, and applications.

Black rice is considered to be functional food containing anthocyanins as bioactive compounds. This study examined the genomic and proteomic patterns in local black rice from Java Island, Indonesia, with attention to the mechanism of anthocyanin synthesis. Three kinds of black rice from Java Island, including black rice from East Java (BREJ), black rice from Central Java (BRCJ), and black rice from West Java (BRWJ), were studied in comparison to white rice (WREJ) and red rice (RREJ). Genomic profiling was done by simple sequence repeat (SSR) analysis, and sequencing of red coleoptile (Rc) and glycosyltransferase (GT) genes, followed by in silico analysis. Total anthocyanin was investigated by ultra-high performance liquid chromatography- diode array detector (UHPLC-DAD). The proteomic profiles were determined by liquid-chromatography and mass spectrometry of tryptic peptides. The SSR profiles showed a specific band in each black rice variant. The Rc gene exon-2 sequences were similar in the three black rice cultivars. The GT gene sequence was identified as a new variant that correlates with the purple stem, leaf, bran, and whole grain morphology seen exclusively in the BRWJ cultivar. The anthocyanin composition in Java black rice is diverse. The highest cyanidin level was seen in BRWJ and the highest level of peonidin-3-O-glucoside in BREJ. Proteomic profiling of the black rice cultivars demonstrated that the expression of proteins that might be related to the levels of anthocyanin synthesis varied. These studies conclude that the genomic, proteomic and anthocyanins composition of Java black rice cultivars may be used the improvement of their functional nutrition values.

Background: One of the main disadvantages of red pigments used in foods is their low extraction yield and storage stability. Roselle flowers are sources of anthocyanins; however, these are unstable during storage, but this could be improved with phenolic compounds, which establish bonds with the anthocyanins. The aim of this work was to identify conditions to improve the extraction efficiency and color stability of Hibiscus sabdariffa extract and, furthermore, to identify the phenolic compounds associated with color stability of roselle., Results: The temperature of extraction (35 and 75 °C), the time of extraction (15 and 60 min), type of acid (acetic and citric), percentage of acid (0.5 and 2.0%) and the water:ethanol ratio (20:80 and 80:20) did not affect the yield of anthocyanins; only the solid:solvent ratio had an effect in the anthocyanin extraction yield. The extraction with 80% ethanol decreased up to 50% the anthocyanin degradation in the extracts stored at 4, 25, 35, 45, and 80 °C. Phenolic acids and flavonoids were quantified by ultrahigh-performance liquid chromatography coupled with triple quadrupole electrospray ionization mass spectrometry. These compounds were analyzed using principal component analysis, and the H. sabdariffa extract, with greater stability, was found to be associated with the presence of quercetin, myricetin, kaempferol 3-O-glucose, ellagic acid, and rutin., Conclusion: H. sabdariffa extract with increased color stability was extracted with a higher proportion of ethanol and the improvement in the color stability was attributed to the co-extraction of phenolic compounds, principally flavonoids that could interact with anthocyanins and stabilize them. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Colored (orange, pink, red, purple, and blue) corn strongly attracted attention on its healthy properties mainly due to its anthocyanin and carotenoid composition which is also responsible for its pigmentation. The present review summarized the recent updates on the extraction and chemical characterization of the main plant secondary metabolites present in colored seeds, kernel, cob, husk, and silk. The main approaches used to stabilize the extracts have been discussed as well as their food and non-food uses. Both in vitro and in vivo (animal models) studies on the different effects (antibacterial, antimutagenic, antioxidant, and anti-inflammatory activities, effects on metabolic syndrome, diabetes, glucose and lipidic metabolism, and neuroprotection) of pigmented extracts on animal and human health have been summarized.

Anthocyanins have attracted great attention because of their potential therapeutic benefit. However, the effective technique for simultaneous separation and preparation multiple anthocyanin monomers with high purity and high yield is still deficient. In this study, the chromatographic conditions of HPLC were optimized to investigate six well-known major anthocyanins (delphinidin-3-O-glucoside, cyanidin-3-O-glucoside, petunidin-3-O-glucoside, pelargonidin-3-O-glucoside, peonidin-3-O-glucoside and malvidin-3-O-glucoside) in blueberry. The separation conditions were optimized in analytical HPLC and further applied in semi-preparative HPLC to prepare anthocyanin monomers. The results showed that six well-known major anthocyanins were well separated under the condition of using acetonitrile-water (contained 0.3% phosphoric acid) as a mobile phase with gradient elution at a detection wavelength of 520 nm. The method showed good linear correlations between the concentrations and peak areas of the six components with correlation coefficients greater than 0.9994, and the detection limits of the six anthocyanins were 0.010-0.035 μg/mL, and the quantification limits were 0.033-0.117 μg/mL, which was suitable for the determination of anthocyanins in products. In the same experimental conditions, six well-known major anthocyanins were simultaneously prepared by semi-preparative HPLC with high purity to 99% and high yield to 22.5%. This study provides a practical and valuable method for simultaneous determination and preparation of six well-known major anthocyanins.

Utilization of economical and environmental methods as an alternative process to recover the industrial crops and food products into high-added value compounds is of great significant. In the current study, microwave assisted extraction (MAE) was optimized by response surface methodology (RSM) in order to evaluate the waste of sour cherry as a source of phenolic compounds rich in anthocyanins. The process parameters (microwave power, irradiation time and ethanol solvent concentration) of MAE method were optimized by face centered composite design of RSM. Responses such as total phenolic componds (TPC), total anthocyanin (TA) contents and antioxidant capacity (DPPH) of extracts were measured spectrophotometrically after extractions of samples. The optimized result of MAE was 500 W of microwave power, 90 s of irradation time and 80% ethanol solvent concentration. Antioxidant capacity was tested using by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical. Chromatographic analysis (HPLC) was also used to measure the concentration of major anthocyanin (cyanidin-3-glucoside) of the samples. Maximum predicted TPC, TA and DPPH yields on optimized conditions were 44.15 mg-GAE/g-FM (mg- gallic acid equivalent per g- fresh matter), 12.47 mg-cyanidin-3-glucoside/g-FM and 69.90 (%, inhibition), respectively. A stability assay under different conditions (light, dark, ambient condition, refrigerator and deep freezer) has been also performed in order to display the stability of bioactivity profile. All of the process parameters were significant at the level of p  < 0.0001.

Sorghum derived 3-deoxyanthocyanin (DXA) pigments are stable relative to their anthocyanin analogs, and are of growing interest in food applications. However, the 3DXA are poorly extractable from grain tissue. This work aimed to determine the relative stability and extractability of sorghum 3-DXA vs anthocyanins from maize and cowpea under microwave-assisted extraction (MAE). UV-Vis and UPLC-MS/MS spectrometry were used to characterize the properties. The 3-DXA remained structurally stable to MAE conditions up to 1200 W/100 °C/30 min. MAE increased sorghum 3-DXA yield 100% versus control (3100 vs 1520 mg/g). On the other hand, both maize and cowpea anthocyanins were unstable and rapidly degraded under MAE. Cell wall-derived ferulate esters were detected in sorghum and maize MAE extracts, indicating cell wall degradation occurred during MAE. Thus the enhanced extraction of 3-DXA under MAE was due to their structural stability, along with improved diffusion from cell matrix due to microwave-induced sorghum cell wall disruption., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Blueberry wine residues produced during the wine-brewing process contain abundant anthocyanins and other bioactive compounds. To extract anthocyanins from blueberry wine residues more efficiently, a novel procedure of ultrasound-assisted deep eutectic solvent extraction (UADESE) was proposed in this work. The extraction process was optimized by response surface methodology coupled with genetic algorithm. The optimum extraction parameters to achieve the highest yield of anthocyanins (9.32 ± 0.08 mg/g) from blueberry wine residues by UADESE were obtained at water content of 29%, ultrasonic power of 380 W, extraction temperature of 55 °C, and extraction time of 40 min. The AB-8 macroporous resin combined with Sephadex LH-20 techniques was used to purify the crude extract (CE) obtained under optimum extraction conditions and analyze the anthocyanins composition by HPLC-ESI-MS/MS. The cyanidin-3-rutinoside with purity of 92.81% was obtained. The HepG2 antitumor activity of CE was better than that of the purified anthocyanins component. Moreover, CE could increase the intracellular reactive oxygen species levels and the apoptosis, and arrest HepG2 cells in the S phases. These findings provided an effective and feasible method for anthocyanins extraction, and reduced the environmental burden of this waste.

The aim of this study was to investigate the effects of different solvent and extraction temperatures on the free and bound phenolic compounds and antioxidant activity of dried blackcurrant skins (DBS). Apart from acetic acid buffer solution, different solvent systems, including water, methanol, and mixtures of methanol/water, were also employed and the effects of solvent and temperature (30 and 50 °C) on the free and bound forms of anthocyanins, hydroxycinnamic acids, and flavonols yield were assessed. The results showed that among all solvents, acetic acid buffer resulted in the highest free anthocyanin content (1,712.3 ± 56.1 mg/100 g) (P < 0.05) after 2 hr extraction at 50 °C from DBS, while lower amounts of bound anthocyanins and anthocyanidins were detected after acid hydrolysis. Acetic acid buffer extracts exhibited the highest free hydroxycinnamic acid content (268.0 ± 4.5 mg/100 g), total phenolic content (3702.2 ± 259.3 mg GAE/100 g), and DPPH activity (60.7 ± 2.0% of inhibition). However, their free flavonol content was slightly lower (60.2 ± 0.8 mg/100 g) compared to 100% methanol at 30 and 50 °C (71.4 ± 1.5 mg/100 g and 71.5 ± 6.2 mg/100 g, respectively). Two-way ANOVA indicated interactions between solvent and temperature (P < 0.05), which suggested that the relationship between solvent and phenolic compounds depends on the extraction temperature. PRACTICAL APPLICATION: Overall, acetic acid buffer is more environmental friendly, efficient, and cost effective than other solvents, thus, offering an improved extraction method for phytochemicals as valuable ingredients for nutraceutical applications, from underutilized dried blackcurrant skins (DBS)., (© 2020 Institute of Food Technologists®.)

Grape skins produced during the grape juice production and processing contain abundant anthocyanins and other active compounds. Consequently, this study optimized the extraction conditions for ultrasound-assisted enzymatic extraction (UAEE) of anthocyanins from grape skins via response surface methodology coupled with genetic algorithm. The optimum extraction parameters to achieve the highest anthocyanins yield (3.01 ± 0.04) mg/g from grape skins by UAEE were obtained under an extraction temperature of 50 °C, ultrasonic power of 400 W, pectinase dosage of 0.16%, and extraction time of 28 min. The AB-8 macroporous resin combined Sephadex LH-20 techniques were further employed to purify the anthocyanins extracts obtained under optimum extraction conditions (AEOEC), and the main anthocyanins were identified using high-performance liquid chromatography tandem mass spectrometry. The purified anthocyanins contained two anthocyanins in terms of delphinidin-3,5-O-diglucoside and cyanidin-3-O-rutinoside with purity of 91.35% and 92.64%, respectively. Ultimately, we further evaluated the antitumor activity of AEOEC and two purified anthocyanins on breast cancer. The results indicated that the antitumor effect of AEOEC on breast cancer MCF-7 cells was better than that of two purified anthocyanins. In addition, AEOEC could memorably increase intracellular reactive oxygen species levels and apoptosis of MCF-7 cells, and arrest MCF-7 cells in the G2/M phases. The findings provide an effective and feasible method for anthocyanins extraction and reduce the environmental burden of this waste., (© 2020 Institute of Food Technologists®.)

Three O-methyl anthocyanidin 3-O-β-D-glucopyranosides were isolated from bilberry extract on a large-scale basis together with two non O-methyl analogues. Anthocyanidin 3-O-β-D-galactopyranosides were removed from bilberry extract together with parts of anthocyanidin 3-O-α-L-arabinopyranosides after treatment with β-galactosidase. The remaining arabinopyranosides were removed by applying acid catalytic hydrolysis. The amounts of anthocyanins recovered as flavylium trifluoroacetic acid salt were as follows: 630 mg for petunidin 3-O-β-D-glucopyranoside, 423 mg for peonidin 3-O-β-D-glucopyranoside, 588 mg for malvidin 3-O-β-D-glucopyranoside, 877 mg for delphinidin 3-O-β-D-glucopyranoside, and 742 mg for cyanidin 3-O-β-D-glucopyranoside.

Obesity is a chronic disease that has been causing serious problems all over the world. However, there is a lack of available therapeutic approaches to treat obesity. The FDA-approved drug orlistat has severe side effects, such as abdominal pain, flatulence and oily stool. As the therapeutic target of orlistat is pancreatic lipase, there is an urgent need for discovery of new pancreatic lipase inhibitors from natural sources that have reduced side effects compared with orlistat. In this study, ultrafiltration in combination with molecular simulation and spectroscopy was reported as an effective approach for identifying new pancreatic lipase inhibitors from anthocyanin-rich berry sources. Using this approach, four monomeric anthocyanins cyanidin-3-O-arabinoside (C3A), cyanidin-3-O-galactoside (C3Ga), peonidin-3-O-arabinoside (Pn3A) and peonidin-3-O-galactoside (Pn3Ga) from cranberries were discovered as potent pancreatic lipase inhibitors. These four cranberry anthocyanins were shown to form hydrophobic interactions and hydrogen bonds with pocket amino acid residues in molecular docking and molecular dynamics simulations. C3A showed greater impact on secondary structures of the enzyme and showed higher binding capacity with the enzyme compared with C3Ga, Pn3A and Pn3Ga as observed by CD and fluorescence spectroscopy. The structure-activity relationships were then investigated and summarized as both the structures of the B ring and glycosyl group were related to the inhibitory activities of anthocyanins. In short, our results suggested that cranberry anthocyanins could be developed as food supplements to facilitate the prevention and treatment of obesity.

Cognitive decline is associated with lifestyle-related factors such as overweight, blood pressure, and dietary composition. Studies have reported beneficial effects of dietary anthocyanins on cognition in older adults and children. However, the effect of anthocyanin-rich Aronia melanocarpa extract (AME) on cognition is unknown. Therefore, this study aimed to determine the effect of long-term supplementation with AME on cognitive performance, mood, and vascular function in healthy, middle-aged, overweight adults. In a randomized double-blind placebo-controlled parallel study, 101 participants either consumed 90 mg AME, 150 mg AME, or placebo for 24 weeks. The grooved pegboard test, number cross-out test, and Stroop test were performed as measures for psychomotor speed, attention, and cognitive flexibility. Mood was evaluated with a visual analogue scale, serum brain-derived neurotrophic factor (BDNF) was determined, and vascular function was assessed by carotid ultrasounds and blood pressure measurements. AME improved psychomotor speed compared to placebo (90 mg AME: change = -3.37; p = 0.009). Furthermore, 150 mg AME decreased brachial diastolic blood pressure compared to 90 mg AME (change = 2.44; p = 0.011), but not compared to placebo. Attention, cognitive flexibility, BDNF, and other vascular parameters were not affected. In conclusion, AME supplementation showed an indication of beneficial effects on cognitive performance and blood pressure in individuals at risk of cognitive decline.

Riceberry rice ( Oryza sativa L.) is a new pigmented variety of rice from Thailand. Despite its high anthocyanin content, its effect on adipogenesis and adipocyte function remains unexplored. We investigated whether Riceberry rice extract (RBE) impacted cell proliferation by examining viability and cell cycle, using preadipocyte 3T3-L1 cells. To test RBE's effect on adipocyte formation, cells were cultured in adipogenic medium supplemented with extract and adipocyte number and triglyceride levels were quantified. Furthermore, Akt1 phosphorylation along with RT-qPCR and intracellular calcium imaging were performed to obtain an insight into its mechanism of action. The effect of RBE on adipocyte function was investigated using glucose uptake and lipolysis assays. Treatment of cells with RBE decreased preadipocyte number without cytotoxicity despite inducing cell cycle arrest ( p < 0.05). During adipogenic differentiation, RBE supplementation reduced adipocyte number and triglyceride accumulation by downregulating transcription factors (e.g., PPARγ, C/EBPα, and C/EBPβ) and their target genes ( p < 0.05). The Akt1 phosphorylation was decreased by RBE but insignificance, however, the extract failed to increase intracellular calcium signals. Finally, the treatment of adipocytes with RBE reduced glucose uptake by downregulating Glut4 mRNA expression and enhanced isoproterenol-induced lipolysis ( p < 0.05). These findings suggest that RBE could potentially be used in the treatment of obesity by inhibiting adipocyte formation and proliferation.

Choline is converted to trimethylamine by gut microbiota and further oxidized to trimethylamine- N -oxide (TMAO) by hepatic flavin monooxygenases. Positive correlation between TMAO and chronic diseases has been reported. Polyphenols in black raspberry (BR), especially anthocyanins, possess various biological activities. The objective of this study was to determine the effects of BR extract on the level of choline-derived metabolites, serum lipid profile, and inflammation markers in rats fed high-fat and high-choline diets. Forty female Sprague-Dawley (SD) rats were randomly divided into four groups and fed for 8 weeks as follows: CON (AIN-93G diet), HF (high-fat diet), HFC (HF + 1.5% choline water), and HFCB (HFC + 0.6% BR extract). Serum levels of TMAO, total cholesterol, and low-density lipoprotein (LDL)-cholesterol and cecal trimethylamine (TMA) level were significantly higher in the HFC than in the HFCB. BR extract decreased mRNA expression of pro-inflammatory genes including nuclear factor-κB (NF-κB), interleukin (IL)-1β, IL-6, and cyclooxygenase-2 (COX-2), and protein expression of NF-κB and COX-2 in liver tissue. These results suggest that consistent intake of BR extract might alleviate hypercholesterolemia and hepatic inflammation induced by excessive choline with a high-fat diet via lowering elevated levels of cecal TMA and serum TMAO in rats.

The present study aimed to screen grape extracts as novel α-glucosidase inhibitors to prevent type-2 diabetes and hyperglycemia. The total polyphenol content (TPC) was measured by Folin-Ciocalteu assay and the stilbene, anthocyanin and flavan-3-ol compounds were measured by Ultra High-Performance Liquid Chromatography coupled to Mass Spectrometry (UHPLC-MS). The α-glucosidase inhibitory of seed and skin Tannat grape extracts at four ripening stages were investigated. The highest TPC values were measured in seeds at the "veraison stage" (65.29 ± 5.33 g of Gallic Acid Equivalent (GAE) per kilogram of Fresh Weight (FW)). This was in accordance with the high flavan-3-ol contents measured for these two extracts (43.22 ± 2.59 and 45.45 ± 6.48 g/kg of seeds FW, respectively). The skin and seed extracts at the first stage of ripening exerted strong α-glucosidase inhibition, exceeding 95% ( p < 0.05). A high linear correlation (R = 0.723, p ≤ 0.05) was observed between flavan-3-ol contents and the α-glucosidase inhibitory activity. The stilbene contents and this activity were moderately to strongly anti-correlated (R = -0.828, p ≤ 0.05 for trans -resveratrol). The enzyme kinetic studies revealed a mixed type of inhibition. This study brings promising results for the therapeutic potential of seed and skin Tannat grape extracts as a functional food product with anti-diabetic activity.

The potential of passion fruit ( Passiflora edulis Sims) epicarp to produce anthocyanin-based colorants with bioactive properties was evaluated. First, a five-level three-factor factorial design coupled with response surface methodology was implemented to optimize the extraction of anthocyanins from dark purple epicarps. The extraction yield and cyanidin-3- O -glucoside content were used as response criteria. The constructed models were fitted to the experimental data and used to calculate the optimal processing conditions ( t = 38 min, T = 20 °C, S = 0% ethanol/water ( v/v ) acidified with citric acid to pH 3, and R S/L = 50 g/L) that lead to maximum responses (3.4 mg/g dried epicarp and 9 mg/g extract). Then, the antioxidant, antimicrobial, and cytotoxic activities of anthocyanin extracts obtained using the optimized method and a conventional extraction method were evaluated in vitro. The extract obtained by the optimized method revealed a higher bioactivity, in agreement with the higher cyanidin-3- O -glucoside content. This study highlighted the coloring and bioactive potential of a bio-based ingredient recycled from a bio-waste, which promotes a sustainable bioeconomy in the agri-food sector.

Anthocyanins are a bioactive compound belonging to the flavonoid classthatis present in human nutrition through plant-based foods. Due to their antioxidant properties, several health benefits related to their consumption are reported in the literature. The stability of the color and the properties of anthocyanins is strongly affected by pH, solvent, temperature, and other environmental conditions. In addition, the insufficient residence time of anthocyanins in the upper digestive tract causes apartialabsorption, which needs to be improved. These factshave led researchers to investigate new forms of processing that provide minimal degradation. Microencapsulation is a promising possibility to stabilize anthocyanin extracts and allow their addition to food products in a more stable form. The microcapsules can still provide a prolonged gastrointestinal retention time caused by the improvement of the bioadhesive properties in the mucus covering the intestinal epithelium. Although there are efficient and emerging techniques, anthocyanins microencapsulation is still a challenge for the food industry. The purpose of this work is to provide an overview of anthocyanins structure, absorptionand protection, and to show the main conventional and emerging microencapsulation methods and their pros and cons., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

This study aimed to investigate the beneficial effects of A. melanocarpa on testosterone propionate (TP)-induced benign prostatic hyperplasia (BPH) in Wistar rats. Moreover, the bioactive constituents in the extract were determined using LC/MS and HPLC analyses. The dried fruits of A. melanocarpa were extracted using accelerated solvent extraction (ASE) under different extract conditions (temperature, 30 C or 100 C; extract solvent, 60% or 100% ethanol) to yield four extracts (T1~T4). Of the four A. melanocarpa extracts, T1 extracted under the condition of 100% ethanol/low temperature (30 C) exhibited the greatest inhibitory activity on TP-induced prostatic hyperplasia in rats. The administration of T1 (100 mg/kg body weight, p.o.) for six weeks attenuated TP-induced prostate enlargement and reduced the levels of dihydrotestosterone (DHT) and 5α-reductase in both serum and prostate tissue. The suppression of PCNA mRNA expression in prostate tissue was remarkable in T1-treated rats. In LC/MS analysis, the levels of main anthocyanins and phenolics were significantly higher in T1 than in the other extracts. Furthermore, the quantitative study showed that the contents of cyanidin-3-glucose and cyanidin-3-xylose in T1 exhibited 1.27~1.67 and 1.10~1.26 folds higher compared to those in the other extracts. These findings demonstrated that A. melanocarpa extract containing anthocyanins as bioactive constituents attenuated the development of testosterone-induced prostatic hyperplasia, and suggested that this extract has therapeutic potential to treat prostate enlargement and BPH., Competing Interests: The authors declare no conflicts of interest.

Delphinidin-3-rutinoside, a high-value of anthocyanin, was isolated and purified by ionic liquid (IL)-modified countercurrent chromatography (CCC) from waste peel of eggplant (Solanum melongena), one of the most common vegetables consumed all around the world. Different conventional CCC and IL-CCC solvent systems were evaluated in respect of partition coefficient (K), separation factor (α), and stationary phase retention factor (S f ) to separate polar target and other components. Basic solvent system, kind of ILs, and amount of ILs were systematically optimized by totally K-targeted strategy, which drastically reduced the experimental effort. Finally, a novel CCC two-phase solvent system (methyl tert-butyl ether-butanol-acetonitrile-1% trifluoroacetic acid water-1-butyl-3-methylimidazolium hexafluorophosphate ([C 4 MIM][PF 6 ]) [2:4:1:5:0.2; v/v/v/v/v]) was successfully established and applied. The baseline separation of target fraction was obtained in one cycle process. The purity of delphinidin-3-rutinoside was over 99%. Moreover, the distribution behavior of different kinds of ILs in biphasic solvent system and the removal method of ILs were explored. The results showed that hydrophobic IL significantly improved the partition of polar anthocyanin in organic solvent system, thereby the separation resolution and stationary phase retention through introducing intermolecular forces. This IL-modified CCC strategy may be applied for the separation of other anthocyanins from variety of natural food resources and waste., (© 2020 Institute of Food Technologists®.)

The aim of this study was to extract and purify anthocyanins from Lycium ruthenicum Murr. and evaluate their tyrosinase inhibitory activity. Response surface methodology was devoted to optimize enzyme-assisted extraction of anthocyanins from L. ruthenicum dried fruits. Extraction at 38 °C for 37 min using water-containing pectinase (52.04 mg/100 g dried fruit) rendered an anthocyanin extraction yield of 19.51 ± 0.21 mg/g. The purified anthocyanins were separated from the extract by macroporous resin XDA-6. Antioxidant tests in vitro suggested that the extract and the purified anthocyanins exhibited a potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity, 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging capacity, hydroxyl radical scavenging capacity, superoxide radical scavenging capacity, and total reducing power. Thirteen anthocyanins from L. ruthenicum dried fruits were analyzed by HPLC-MS. Moreover, the purified anthocyanins had inhibitory effect on tyrosinase monophenolase (IC 50 = 1.483 ± 0.058 mg/mL), and the type of inhibition was competitive inhibition (K i = 39.83 ± 1.4 mg/mL). The maximum inhibitory activity of the purified anthocyanins (3.00 mg/mL) on tyrosinase diphenolase was 42.16 ± 0.77%, and the type of inhibition was anticompetitive inhibition (K is = 2.387 ± 0.10 mg/mL). PRACTICAL APPLICATION: The anthocyanins from L. ruthenicum dried fruits can be used as tyrosinase inhibitors in medicine, cosmetics, and food preservation industries., (© 2020 Institute of Food Technologists®.)

Anthocyanins in wine principally depend on grape skin extractable anthocyanin content, that is, the amount of anthocyanins present in grape skin that are released to wine during the maceration stage. This amount of extractable anthocyanins is closely linked to the cell wall degradation of skin cells. Indeed, among other methodologies, the maceration in presence of different enzymes can be used to increase cell wall degradation, and therefore, the amount of anthocyanins extracted from grape skins to wine. Vitis vinifera L. cv. Tempranillo and Syrah red grapes have been identified as samples with low anthocyanin extraction potential by near infrared hyperspectral imaging. Grape skins have been macerated in the presence of cellulase, glucosidase, and pectinase. Then, color of the supernatants and phenolic compounds extracted from grape skins (total phenols, total flavanols, and total and individual anthocyanins) has been determined. Cellulase and glucosidase have shown a positive effect in the extraction of phenolic compounds from these grapes. Macerations carried out in the presence of cellulase have produced supernatants with a more intense color (lower lightness and higher chroma values), and a higher extraction of flavanols and anthocyanins than the respective control essays. However, pectinase treatments have produced the opposite effect, which could be partially explained by an eventual interaction between the cell wall polysaccharides liberated by pectinase and the phenolic compounds extracted. Synergy effects do not appear between cellulase and glucosidase. Moreover, the negative effect of the addition of pectinase might be due to the interactions between the cell wall material liberated by pectinase and the phenolic compounds extracted. PRACTICAL APPLICATION: In the present study, grape samples with a low anthocyanin extraction potential have been identified, and these samples have been macerated in the presence of different enzymes. The applied enzymes were three of the most common enzymes that are applied in the wine industry. Individual enzymes and mixtures have been applied to Syrah and Tempranillo grape skin samples and the results have been compared to control macerations. Knowledge in this topic will help the production of quality wines., (© 2020 Institute of Food Technologists®.)