Crucial Challenges in the Development of Green Extraction Technologies to Obtain Antioxidant Bioactive Compounds from Agro-industrial By–products

Exploitation of agro-industrial by-products represents an important source of bioactive compounds that can be used both directly as ingredients and for the production of functional ingredients. Among these compounds, polyphenols are capable of strengthening endogenous antioxidant defences in human tissues, preventing cardiovascular and neurodegenerative diseases. The present paper aims to evaluate and review various green extraction technologies for a cheap, fast, eco-friendly procedure to obtain these bioactive molecules. Several physicochemical approaches can be used with the aim of optimizing the use of energy, solvents, and pressure; among them are ultrasound-assisted extraction, subcritical and supercritical fluid extraction, extraction with neoteric solvents (ionic liquids, deep eutectic solvents, and natural deep eutectic solvents), microwave-assisted extraction, pressurized liquid extraction, pulsed electric field, multi-frequency multimode modulated technology, rapid solid liquid dynamic extraction, and enzyme-assisted extraction. The challenges and future work regarding the development of these green products for the commercial markets were comprehensively evaluated. This work is licensed under a Creative Commons Attribution 4.0 International License

Composition of Sulla (Hedysarum coronarium L.) honey solvent extractives determined by GC/MS: norisoprenoids and other volatile organic compounds

Samples of unifloral sulla (Hedysarum coronarum L.) honey from Sardinia (Italy) were analysed. To investigate the chemical composition of the honey volatiles two solvent systems were used for ultrasonic solvent extraction (USE): 1) a 1:2 (v/v) pentane and diethyl ether mixture and 2) dichloromethane. All the extracts were analysed by GC and GC/MS. These procedures have permitted the identification of 56 compounds that include norisoprenoids, benzene derivatives, aliphatic compounds and Maillard reaction products. Norisoprenoids were the major compounds in both extracts, dominated by vomifoliol (5.3-11.2%; 9.6-14.0%) followed by minor percentages of other norisoprenoids such as α-isophorone, 4-ketoisophorone, 3-oxo-α-ionol or 3-oxo-α-ionone. Other abundant single compounds in the extracts were 3-hydroxy-4-phenylbutan-2-one (0.8-5.4%; 0.6-5.7%) and methyl syringate (3.0-5.7%; 2.2-4.1%). The composition of the volatiles and semi-volatiles in the obtained extracts suggests that sulla honey is quite distinctive relative to the other honeys that have been chemically studied by GC/MS, but no specific markers of the honey botanical origin were foun

Analysis of polyphenolic composition and stability of magistral preparation based on salviae officinalis folium

A magistral preparation based on the sage leaf decoction or infusion is one of the most popular herbal prescriptions prepared in pharmacies in Lower Silesia (Poland) for gargling in the case of pharyngitis or mouth infections. Other components of this preparation are boric acid, aluminum(III) acetotartrate or Burow's solution, and glycerol. The study aimed to investigate the polyphenolic composition and relations between herbal and chemical ingredients that are present in this mixture in comparison with the traditional aqueous galenic forms - infusion and decoction. The analysis was performed using high-performance liquid chromatography with diode-array detector (HPLC-DAD) and electrospray ionization mass spectrometry (ESI-MS) methods. The main polyphenolic ingredients of sage leaf were rosmarinic acid (RA) and luteolin 7-O-β-glucuronide. The RA predominance was observed in the infusion, decoction and magistral preparation. Comparing the contents of analyzed polyphenols, it was confirmed that their levels in the decoction were higher than in the infusion. Thus, the pharmacological activity of the examined magistral preparation results from the presence of boric acid, aluminum(III) salts and polyphenolic components of sage leaf hot water extract. The succeeding study showed that in the presence of excess salts of aluminum(III) and boric acid, sage polyphenols are partially soluble in an aqueous medium (mainly phenolic acids) and complexed as precipitated sediment (especially flavonoids). The aqueous solutions of this preparation obtained according to the recommendation are cloudy, which suggests limited solubility of the formed complexes. Therefore, the therapeutic activity should be associated with the presence of water-soluble caffeic acid esters like rosmarinic acid. The influence of luteolin glycosides coordination complexes is not evident. The obtained results also confirm the stability of the examined prescription formulation in the conditions of cold storage (4-6OC) within 7 days from its production

Investigation into Polyphenol Profile and Biological Activities of Enriched Persimmon/Apple Smoothies during Storage

Smoothies are becoming an increasingly popular product as a healthy alternative to snacks. The consumer expects from this product that, apart from its nutritional value, it will also be qualitatively stable during storage. Therefore, in this study, original smoothies obtained with persimmon fruit puree and apple juice (Dk/Md) enriched with Arbutus unedo fruits, Myrtus communis purple berry extract, Acca sellowiana, and Crocus sativus petal juice were evaluated for their polyphenol composition, antioxidant activity, and inhibition on targeted digestive enzymes, over six months of storage. The amount of polyphenols evaluated by UPLC-PDA analysis decreased in six months from 23.5% for both Dk/Md and enriched C. sativus smoothies to 42.5% for enriched A. sellowiana, with anthocyanins the most sensitive compounds (71.7–100% loss). Values of antioxidant assays generally strongly decreased during the first three months (up to ca. 60%) and to a lesser extent in the following three months (0.4–27%). In addition, inhibitory activity on α-amylase, α-glucosidase, and pancreatic lipase, especially on the last two enzymes, was negatively affected by time storage. The outcome of this study indicates that persimmon fruit is a good option for producing smoothies, and enrichment with other plant extracts can enhance the bioactive compound content and biological activities. It is recommended that appropriate storage strategies to preserve the properties of those smoothies should be developed

Thymol as a Component of Chitosan Systems—Several New Applications in Medicine: A Comprehensive Review

Thymol, a plant-derived monoterpene phenol known for its broad biological activity, has often been incorporated into chitosan-based biomaterials to enhance therapeutic efficacy. Using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines, we conducted a systematic literature review from 2018 to 2023, focusing on the biomedical implications of thymol-loaded chitosan systems. A review of databases, including PubMed, Scopus, and Web of Science was conducted using specific keywords and search criteria. Of the 90 articles, 12 were selected for the review. Thymol-loaded chitosan-based nanogels (TLCBS) showed improved antimicrobial properties, especially against multidrug-resistant bacterial antagonists. Innovations such as bipolymer nanocarriers and thymol impregnated with photosensitive chitosan micelles offer advanced bactericidal strategies and show potential for bone tissue regeneration and wound healing. The incorporation of thymol also improved drug delivery efficiency and biomechanical strength, especially when combined with poly(dimethylsiloxane) in chitosan–gelatin films. Thymol–chitosan combinations have also shown promising applications in oral delivery and periodontal treatment. This review highlights the synergy between thymol and chitosan in these products, which greatly enhances their therapeutic efficacy and highlights the novel use of essential oil components. It also highlights the novelty of the studies conducted, as well as their limitations and possible directions for the development of integrated substances of plant and animal origin in modern and advanced medical applications

Exploring Phenolic Compounds Extraction from Saffron (C. sativus) Floral By-Products Using Ultrasound-Assisted Extraction, Deep Eutectic Solvent Extraction, and Subcritical Water Extraction

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

Qualitative profile and quantitative determination of flavonoids from Crocus sativus L. petals by LC-MS/MS

From the methanolic extract of Crocus sativus petals nine known flavonoids have been isolated and identified, including glycosidic derivatives of quercetin and kaempferol as major compounds (1–2), and their methoxylated and acetylated derivatives. Additionally, LC-ESI-MS qualitative and LC-ESI-MS/MS quantitative studies of the major compounds of the methanolic extract were performed. The high content of glycosylated flavonoids could give value to C. sativus petals, which are a waste product in the production of the spice saffron

Screening of Satureja subspicata Vis. honey by HPLC-DAD, GC-FID/MS and UV/VIS: prephenate derivatives as biomarkers

The samples of Satureja subspicata Vis. honey were confirmed to be unifloral by melissopalynological analysis with the characteristic pollen share from 36% to 71%. Bioprospecting of the samples was performed by HPLC-DAD, GC-FID/MS, and UV/VIS. Prephenate derivatives were shown to be dominant by the HPLC-DAD analysis, particularly phenylalanine (167.8 mg/kg) and methyl syringate (MSYR, 114.1 mg/kg), followed by tyrosine and benzoic acid. Higher amounts of MSYR (3-4 times) can be pointed out for distinguishing S. subspicata Vis. honey from other Satureja spp. honey types. GC-FID/MS analysis of ultrasonic solvent extracts of the samples revealed MSYR (46.68%, solvent pentane/Et2O 1:2 (v/v); 52.98%, solvent CH2Cl2) and minor abundance of other volatile prephenate derivatives, as well as higher aliphatic compounds characteristic of the comb environment. Two combined extracts (according to the solvents) of all samples were evaluated for their antioxidant properties by FRAP and DPPH assay; the combined extracts demonstrated higher activity (at lower concentrations) in comparison with the average honey sample. UV/VIS analysis of the samples was applied for determination of CIE Lab colour coordinates, total phenolics (425.38 mg GAE/kg), and antioxidant properties (4.26 mmol Fe2+/kg (FRAP assay) and 0.8 mmol TEAC/kg (DDPH assay)

GC-MS Fingerprints and Other Physico-chemical Characteristics of Rare Unifloral Prunus cerasus L. Honey

GC-MS fingerprints of unifloral sour cherry ( Prunus cerasus L.) honey were investigated for the first time by GC-FID and GC-MS {after headspace solid phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE)}. Additionally, other physico-chemical characteristics of the samples were determined (total phenolic content, antioxidant activity and CIE L*a*b*C*h chromatic coordinates). The principal volatile components of the honey headspace were lilac aldehydes (46.0; 50.6%) along with benzaldehyde (18.0; 19.4%). The dominant component of the dichloromethane USE extract was vomifoliol (39.6; 44.9%). The abundant identified compounds may only serve as non-specific markers of the honey's botanical origin since they also occur in other honey types. The honey contained low-moderate amount of polyphenols (209.0 - 309.5 mg GAE/kg) and exhibited moderate antioxidant activity (0.4 - 0.6 mmol TEAC/kg; 1.6 - 1.9 mmol Fe2+/kg)

Volatile profile, phytochemicals and antioxidant activity of virgin olive oils from Croatian autochthonous varieties Mašnjača and Krvavica in comparison with Italian variety Leccino

Virgin olive oils (VOOs) obtained from the fruits of Croatian autochthonous varieties Mašnjača and Krvavica were extensively characterized for the first time. Investigated oils were compared with the oil obtained from Italian variety Leccino, grown and processed under the same conditions. Headspace volatile profile, tocopherols, chlorophylls, carotenoids and total phenolic content, peroxide value, % acidity, K232, K270 as well as antioxidant activity (DPPH) of the oils’ hydrophilic fractions (HFs) including their phenolic composition were assessed by means of HS-SPME/GC-MS, HPLC-FL, HPLC-DAD and spectrophotometric methods, respectively. Most of the studied quality parameters varied between the cultivars. The main volatile compounds detected in all tested olive oils were the C6 compounds derived from polyunsaturated fatty acids through the lipoxygenase pathway. Krvavica oil was characterized by hexanal (8.8%–9.4%). Leccino oil contained the highest percentage of (E)-hex-2-enal (73.4%–74.0%), whereas (Z)-hex-3-enal (21.9%–25.0%) and (E)-hex-2-enal (27.6%–28.9%) dominated in Mašnjača oil. Leccino oil contained the highest amount of tocopherols (312.4 mg/kg), chlorophylls (7.3 mg/kg), carotenoids (4.2 mg/kg) and total phenols (246.6 mg/kg). The HF of Leccino oil showed the highest antioxidant capacity (1.3 mmol TEAC/kg), while the HFs of Mašnjača and Krvavica oils exhibited the activity of 0.5 mmol TEAC/kg