Simultaneous High-Performance Recovery and Extended Acid-Catalyzed Hydrolysis of Oleuropein and Flavonoid Glycosides of Olive (Olea europaea) Leaves: Hydrothermal versus Ethanol Organosolv Treatment.

Olive leaves (OLLs) are an exceptional bioresource of natural polyphenols with proven antioxidant activity, yet the applicability of OLL extracts is constrained by the relatively high polarity of the major polyphenols, which occur as glycosides. To overcome this limitation, OLLs were subjected to both hydrothermal and ethanol organosolv treatments, fostered by acid catalysis to solicit in parallel increased polyphenol recovery and polyphenol modification into simpler, lower-polarity substances. After an initial screening of natural organic acids, oxalic acid (OxAc) was found to be the highest-performing catalyst. The extraction behavior using OxAc-catalyzed hydrothermal and ethanol organosolv treatments was appraised using kinetics, while treatment optimization was accomplished by deploying response-surface methodology. The comparative assessment of the composition extracts produced under optimal conditions of residence time and temperature was performed with liquid chromatography-tandem mass spectrometry and revealed that OLLs treated with 50% ethanol/1.5% HCl suffered extensive oleuropein and flavone glycoside hydrolysis, affording almost 23.4 mg hydroxytyrosol and 2 mg luteolin per g dry weight. On the other hand, hydrothermal treatment with 5% OxAc provided 20.2 and 0.12 mg of hydroxytyrosol and luteolin, respectively. Apigenin was in all cases a minor extract constituent. The study presented herein demonstrated for the first time the usefulness of using a natural, food-grade organic acid to perform such a task, yet further investigation is needed to maximize the desired effect.

Hydrothermal Treatment of Wheat Bran under Mild Acidic or Alkaline Conditions for Enhanced Polyphenol Recovery and Antioxidant Activity.

This study was undertaken to investigate the effects of hydrothermal treatments under mild acid and alkaline conditions on polyphenol release and recovery from wheat bran (WB). After an initial screening of various food-grade substances, strong evidence was raised regarding the potency of citric acid and sodium carbonate to provide WB extracts exceptionally enriched in polyphenols. Thus, these two catalysts were tested under various time and temperature combinations, and the processes were described by linear models based on severity factor. The most effective treatments were those performed with 10% of either citric acid or sodium carbonate, at a constant temperature of 90 °C for 24 h, providing yields in total polyphenols of 23.76 and 23.60 mg g-1 dry mass of ferulic acid equivalents, respectively. Liquid chromatography-mass spectrometry analyses revealed that, while the sodium carbonate treatment afforded extracts enriched in ferulic acid, treatments with citric acid gave extracts enriched in a ferulate pentose ester. The extracts produced from those treatments also exhibited diversified antioxidant characteristics, a fact ascribed to the different polyphenolic composition. To the best of the authors' knowledge, this is the first report demonstrating the effective release of ferulic acid and a ferulate pentose ester from WB, using benign acid and alkali catalysts, such as citric acid and sodium carbonate.

White rice enrichment with phenols upon cooking in olive leaf infusion: a preliminary study.

BACKGROUND: White rice is poor in health-promoting phytochemicals; therefore, the production of a phenol-enriched commodity is highly desirable. Recent findings on its enrichment via cooking in plant extracts are promising, yet studies employing aqueous extracts of olive leaves (OLs), containing well-recognized bioactive phenols (e.g. oleuropein) are absent. In addition, little is known about the levels of phenols that are maintained after rice drying and rehydration, an important aspect for the future design of 'ready-to-eat' functional rice. RESULTS: The examination, for the first time, of white rice adsorption capacity of phenols from OLs upon cooking in infusions containing different levels of phenols, after freeze-drying and rehydration, showed the following: (i) the total phenol content, the antioxidant activity (assessed via 2,2-diphenyl-1-picrylhydrazyl radical and ferric reducing antioxidant power assays), the oleuropein and luteolin-7-O-glucoside levels increased dose dependently; (ii) upon rehydration, the average decrease of total phenol content and antioxidant activity values was significantly lower when an exact volume of water was used compared with an excess (~10% versus 63%). A similar trend was observed for oleuropein (36% versus 83%) and the luteolin-7-O-glucoside (24 versus 82%) levels; (iii) the dried enriched kernels were less bright with a hay-yellow hue (CIELab coordinates). CONCLUSION: White rice enrichment with biophenols from OLs, a by-product of olive tree cultivation, was successful using a simple approach. Despite leaching upon freeze-drying/rehydration, sufficient amounts were maintained to obtain a functional rice that could serve as an alternative dietary source of OLs phenols to non-traditional olive tree product consumers or those refraining from sodium and fats. © 2023 Society of Chemical Industry.

Physicochemical Characterization and Biological Properties of Pine Honey Produced across Greece.

Pine honey is a honeydew honey produced in the East Mediterranean region (Greece and Turkey) from the secretions of the plant sucking insect Marchalina hellenica (Gennadius) (Coccoidea: Marchalini-dae) feeding on living parts of Pinus species. Nowadays, honeydew honey has attracted great attention due to its biological activities. The aim of this study was to study unifloral pine honey samples produced in Greece regarding their physicochemical parameters and antioxidant and antibacterial activity against five nosocomial and foodborne pathogens. These honeys showed physicochemical and microscopic characteristics within the legal limits, except for diastase activity, a parameter known to be highly variable, depending on various factors. Substantially higher levels of H2O2 were estimated compared to other types of honeydew honey, whereas protein content was similar. The total phenolic content was 451.38 ± 120.38 mg GAE/kg and antiradical activity ranged from 42.43 to 79.33%, while FRAP values (1.87 to 9.43 mmol Fe+2/kg) were in general higher than those reported in the literature. Various correlations could be identified among these parameters. This is the first attempt to investigate in depth the antibacterial activity of pine honey from Greece and correlate it with honey quality parameters. All tested honeys exerted variable but significant antibacterial activity, expressed as MIC and MBC values, comparable or even superior to manuka honey for some tested samples. Although honey antibacterial activity is mainly attributed to hydrogen peroxide and proteins in some cases (demonstrated by elevated MICs after catalase and Proteinase K treatment, respectively), no strong correlation between the antibacterial activity and hydrogen peroxide concentration or total protein content was demonstrated in this study. However, there was a statistically significant correlation of moisture, antioxidant and antibacterial activity against Klebsiella pneuomoniae, as well as antioxidant and antibacterial activity against Salmonella ser. Typhimurium. Interestingly, a statistically significant negative correlation has been observed between diastase activity and Staphylococcus aureus antibacterial activity. Overall, our data indicate multiple mechanisms of antibacterial activity exerted by pine honey.

Threatened Cretan species Chaerophyllum creticum Boiss. & Heldr.: phenolic profile by HPLC-PDA-MS and in vitro antioxidant capacity.

In this study, the chemical content of the threatened Cretan endemic Chaerophyllum creticum Boiss. & Heldr. was investigated by High Performance Liquid chromatography-Photodiode Array-Mass Spectrometry (HPLC-PDA-MS) analysis. Leaves, flowers and stems of C. creticum, were extracted via maceration and were assessed for their polyphenolic composition and antioxidant capacity. The highest extraction yields were achieved by methanol and methanol/water. A total of 17 compounds were characterised in C. creticum with luteolin-7-O-glucoside being the predominant glucoside found in all the extracts. Malonic esters were present in all the extracts. The main flavonoids and phenolics were quantified by HPLC-UV in parallel to standard spectrophotometric assays which were used for the determination of the Total Polyphenol content and the Total Flavonoid Content. The antioxidant activity was assessed by two different tests: 2,2-diphenyl-1-picrylhydrazyl (DPPH•) free radical assay and the ferric reducing antioxidant power (FRAP) assay. This is the first report on the chemical content of the Cretan endemic C. creticum.

Quantification of Hydrogen Peroxide in Cretan Honey and Correlation with Physicochemical Parameters.

The aim of the present study is to quantify hydrogen peroxide, generated from various types of honey produced in Crete, as a potent antimicrobial agent, and establish any correlation with their physicochemical parameters. The basic physicochemical parameters (diastase activity, HMF content, moisture, electrical conductivity, color, and sugars) of 30 authentic honey samples were determined. The concentration of hydrogen peroxide in all samples was found to be within the range 0.010-0.092 mM. The known correlation between the electrical conductivity and the color of honey was confirmed in this study. Univariate and multivariate statistics applied to the results indicate that the results can be used to discriminate honey sample groups of different botanical origins.

A Green Extraction Process for Polyphenols from Elderberry (Sambucus nigra) Flowers Using Deep Eutectic Solvent and Ultrasound-Assisted Pretreatment.

Sambucus nigra flowers, known as elderberry flowers (EBF), are a plant tissue rich in polyphenolic phytochemicals with important bioactivities. However, there are few studies dealing with the production of polyphenol-containing EBF extracts. The objective of the investigation presented herein was the development of a high-performance green extraction methodology, to generate EBF extracts enriched in polyphenolic substances, using an efficient deep eutectic solvent, combined with ultrasonication pretreatment. The DES was composed of L-lactic acid (hydrogen bond donor-HBD) and glycine (hydrogen bond acceptor-HBA) and, after an initial screening to properly regulate HBD/HBA ratio, the extraction was optimized by deploying response surface methodology. Under the optimized conditions, which were DES/water (85% w/v), liquid-to-solid ratio 60 mL g-1, and stirring speed 200 rounds per minute, the extraction yield in total polyphenols amounted to 121.24 ± 8.77 mg gallic acid equivalents per g dry matter. The integration of ultrasonication prior to the batch stirred-tank extraction boosted polyphenol recovery of up to 174.73 ± 2.62 mg gallic acid equivalents per g dry matter. Liquid chromatography-mass spectrometry analysis showed that the richest EBF extract obtained was dominated by rutin, a di-p-coumaroylquic acid and chlorogenic acid.

Saffron Processing Wastes as a Bioresource of High-Value Added Compounds: Development of a Green Extraction Process for Polyphenol Recovery Using a Natural Deep Eutectic Solvent.

The current investigation was undertaken to examine saffron processing waste (SPW) as a bioresource, which could be valorized to produce extracts rich in antioxidant polyphenols, using a green, natural deep eutectic solvent (DES). Initially, there was an appraisal of the molar ratio of hydrogen bond donor/hydrogen bond acceptor in order to come up with the most efficient DES composed of L-lactic acid/glycine (5:1). The following step was the optimization of the extraction process using response surface methodology. The optimal conditions thus determined were a DES concentration of 55% (w/v), a liquid-to-solid ratio of 60 mL g-1, and a stirring speed of 800 rounds per minute. Under these conditions, the extraction yield in total polyphenols achieved was 132.43 ± 10.63 mg gallic acid equivalents per g of dry mass. The temperature assay performed within a range of 23 to 80 °C, suggested that extracts displayed maximum yield and antioxidant activity at 50-60 °C. Liquid chromatography-mass spectrometry analysis of the SPW extract obtained under optimal conditions showed that the predominant flavonol was kaempferol 3-O-sophoroside and the major anthocyanin delphinidin 3,5-di-O-glucoside. The results indicated that SPW extraction with the DES used is a green and efficient methodology and may afford extracts rich flavonols and anthocyanins, which are considered to be powerful antioxidants.

Natural food colourants derived from onion wastes: application in a yoghurt product.

The valorization of onion (Allium cepa) solid wastes, a 450,000 tonnes/year waste in Europe, by a green extraction method is presented. Polyphenols of onion solid wastes were extracted using eco-friendly solvents, such as water and glycerol. The 2-hydroxypropyl-ß-cyclodextrin was also used as a co-solvent for the augmentation of the extraction yield. The process has been optimized by implementing a central composite face centered design of experiments, with two replicates in the central point, taking into consideration the following independent variables: glycerol concentration, cyclodextrin concentration and temperature. The assessment of the extraction model was based on two responses: the total pigment yield and the antiradical capacity. LC-MS analysis was also employed in order to identify polyphenols and colourants of the obtained extracts. The main polyphenols found were quercetin and quercetin derivatives and the main colourant was cyanidin 3-O-glucoside. The extract was also tested as a food colourant in a yoghurt matrix. The onion leaf extract was found to be a stable natural colourant and could be utilized as an alternative ingredient to synthetic coloring agents. This article is protected by copyright. All rights reserved.

Characterisation of Polyphenol-Containing Extracts from Stachys mucronata and Evaluation of Their Antiradical Activity.

Background: The aromatic plant Stachys mucronata (Lamiaceae) is endemic to the island of Crete (southern Greece), but as opposed to other native Greek members of this family, this species has never been investigated in the past with regard to its polyphenolic composition and antioxidant potency. Methods: Aerial parts of S. mucronata were exhaustively extracted and partly fractionated through partition, using n-butanol and dichloromethane. Results: Following an initial examination, which consisted of estimating the total polyphenol content and the antiradical activity, the n-butanol extract was found to be by far the richest in polyphenols, exhibiting much stronger antiradical activity compared with the dichloromethane counterpart. On this basis, the n-butanol extract was analysed by liquid chromatography-diode array-mass spectrometry, to tentatively characterise the principal polyphenolic components, which were shown to be flavonol but mainly flavone derivatives. Conclusions: The most potent radical-scavenging compounds were detected in the n-butanol fraction of the extracts, suggesting that the most active antioxidants in S. mucronate are relatively polar. The analyses suggested the major constituents to be derivatives of the flavone luteolin, accompanied by apigenin analogues, as well as flavonol glycosides and chlorogenate conjugates.

Phytochemical analysis and antioxidant activity of Lycium barbarum (Goji) cultivated in Greece.

CONTEXT: The fruit of Lycium barbarum L. (Solanaceae), known as goji berry, has been exploited for a long time in traditional Chinese medicine. In recent decades, it has received much attention as one of the trendiest functional foods with a wide array of pharmacological activities in Western diets. OBJECTIVE: In this study the phenolic profile and potential antioxidant capacity of Lycium barbarum cultivated in Crete (Greece) were investigated. MATERIALS AND METHODS: The berries were defatted with hexane and then extracted with dichloromethane and methanol using a Soxhlet apparatus. Furthermore, the methanol extract was fractionated with ethyl acetate and butanol. All fractions/extracts were tested for their antioxidant activity (DPPH, FRAP, chemiluminescence). Folin-Ciocalteu and LC-DAD-MS analyses were utilized for the identification of the phenolic compounds. RESULTS: The total phenolic content ranged from 14.13 ± 0.40 (water fraction) to 109.72 ± 4.09 (ethyl acetate fraction) mg gallic acid equivalent/g dry extract. Ethyl acetate extract exhibited the highest scavenging activities determined as EC50 (4.73 ± 0.20 mg/mL) and IC50 (0.47 ± 0.001 mg/mL) using DPPH and chemiluminescence assays. Seventeen phenolic compounds, including cinnamoylquinic acids and derivatives, hydrocinnamic acids and flavonoid derivatives, were tentatively identified. To the best of our knowledge, quercetin 3-O-hexose coumaric ester and quercetin 3-O-hexose-O-hexose-O-rhamnose are reported for the first time in goji berry fruits. DISCUSSION AND CONCLUSION: The results of this study suggest that consumption of goji berry fruits could serve as a potential source of natural antioxidant compounds and that goji berry phenolic extracts could be exploited for nutritional pharmaceutical purposes.

Optimization of a green extraction method for the recovery of polyphenols from olive leaf using cyclodextrins and glycerin as co-solvents.

Olive leaf, an agricultural by-product, was studied for the valorization of its biophenols using green extraction techniques; i.e. non-toxic and eco-friendly extraction solvents were used, involving water and glycerol. 2-hydroxypropyl-ß-cyclodextrin (CD), was also employed as an enhancer of the extraction, since cyclodextrins (CD's) are known to improve the extractability of olive leaf polyphenols by forming water soluble inclusion complexes. The process was optimized by implementing a central composite (Box-Behnken) experimental design and response surface methodology, taking into consideration the following independent variables: glycerol concentration (Cgl), CD concentration (CCD) and temperature (T). The evaluation of the extraction model was based on two responses: the total polyphenol yield (YTP) and the antiradical activity (AAR). Optimum values for the extraction process were obtained at 60% (w/v) glycerol content, T = 60 °C and 7% (w/v) CD content. LC-MS analysis was also applied in order to characterize the polyphenolic composition of extracts containing cyclodextrins. The main polyphenols present were oleuropein and oleuropein derivatives. Olive leaf aqueous extracts containing glycerol and cyclodextrins may be used as raw materials/ingredients for several end-users in the food, cosmetic and pharmaceutical industries.