Screening of Bioactive Compounds from Rosa canina L. Peel and Seed Herbal Dust Using Eco-Friendly Extraction Techniques

This study aimed to determine the effectiveness of different green extraction techniques (GETs) on targeted bioactive compounds from artichoke leaf by-products using deep eutectic solvent extraction (DESE), supercritical CO2 extraction (SCO2E), subcritical water extraction (SWE), and ultrasound-assisted extraction (UAE). Moreover, (HR) LC-ESI-QTOF MS/MS and HPLC-PDA analyses were used to perform qualitative–quantitative analysis on the extracts, enabling the detection of several bioactive compounds, including luteolin, luteolin 7-O-glucoside, luteolin 7-O-rutinoside, apigenin rutinoside, chlorogenic acid, and cynaropicrin as the most representative ones. SWE showed better results than the other GETs (TPC: 23.39 ± 1.87 mg/g of dry plant, dp) and appeared to be the best choice. Regarding UAE, the highest total phenols content (TPC) was obtained with 50:50% v/v ethanol: water (7.22 ± 0.58 mg/g dp). The DES obtained with choline chloride:levulinic acid showed the highest TPC (9.69 ± 0.87 mg/g dp). Meanwhile, SCO2E was a selective technique for the recovery of cynaropicrin (48.33 ± 2.42 mg/g dp). Furthermore, the study examined the antioxidant activity (1.10–8.82 mmol Fe2+/g dp and 3.37–31.12 mmol TEAC/g dp for DPPH• and FRAP, respectively) and total phenols content via Folin–Ciocalteu’s assay (198.32–1433.32 mg GAE/g dp), of which the highest values were detected in the SWE extracts. The relationship among the GETs, antioxidant assays, and compounds detected was evaluated using Principal Component Analysis (PCA). PCA confirmed the strong antioxidant activity of SWE and showed comparable extraction yields for the antioxidant compounds between UAE and DESE. Consequently, GETs selection and extraction parameters optimization can be employed to enrich artichoke leaf by-products’ extracts with targeted bioactive compounds.

Three different brands of Rose hip (Rosa canina L.) tea were extracted with water, ethanol (EtOH), methanol (MeOH), and aqueous mixtures (50%, v/v) by ultrasound-assisted extraction (UAE) and Soxhlet methods. Total phenolic content was determined according to the Folin-Ciocalteu method. The results were presented by means of the extract yields and total phenolic contents, expressed in gallic acid equivalent (GAE) per g of dried matter (DM). The greatest amount of extract observed in tea samples was obtained by UAE through water with the value of 619.37 ± 0.58 mg/g DM. Regarding the phenolic content, the best result was achieved by the Soxhlet method through 50% MeOH mixture (59.69 ± 0.89 mg GAE/g DM), followed by the UAE method with water (48.59 ± 0.29 mg GAE/g DM).

Growing interest in sustainable functional food ingredients has accelerated the search for green extraction methods for bioactive compounds. This study systematically evaluates the use of three emerging green solvents, namely γ-valerolactone (GVL), Cyrene™, and ethyl lactate (EL), as alternatives to conventional solvents for extracting phenolic antioxidants from rose hip (Rosa canina L.) fruit. Using maceration, ultrasound-assisted extraction (UAE), and microwave-assisted extraction (MAE), we compared extraction efficiency, total phenolic content, and antioxidant activity across various solvent systems and techniques. Our results demonstrate that MAE consistently provided the highest extraction yields and phenolic recovery, particularly when using ethanol or ethanol/green solvent mixtures. While pure green solvents showed lower extraction efficiency than ethanol, certain binary mixtures, especially GVL with ethanol, delivered promising results both in phenolic yield and antioxidant activity, without significant interference in standard assays. Additionally, while Cyrene™ consistently yielded low extraction efficiencies and low levels of phenolic compounds, its extracts were unique in exhibiting selectivity and stimulated fibroblast migration in vitro, suggesting additional functional benefits for health applications. Overall, our findings support the practical use of selected green solvents in sustainable extraction protocols for food, nutraceutical, and cosmetic industries.

Ultrasound-assisted and subcritical water extraction techniques for maximal recovery of phenolic compounds from raw ginger herbal dust toward in vitro biological activity investigation

Ultrasound-assisted extraction of polyphenols from rosehip (Rosa canina L.) was investigated. The aim of the study was to examine the extraction parameters such as extraction time (30-90 min), temperature (30-50 °C) and solvent concentration (40-100% ethanol, v/v), and to obtain the best possible combinations of these parameters by using response surface methodology. The experiments were performed according to the face-centred central design. Total phenolic content was expressed in gallic acid equivalent (GAE) per gram of dried matter (DM). The second-order polynomial model gave a satisfactory description of the experimental data. 47.23 mg GAE g(-1) DM was predicted at the optimum operating conditions (40% EtOH composition, 50 °C of temperature, and 81.23 min of extraction time). All of the parameters (ethanol concentration, extraction time and extraction temperature) used in this research have the significant effect on the extraction efficiency of total phenolic content in rosehip extracts (P < 0.05). The solvent concentration was proved to be the most significant effective on the yields obtained by ultrasound-assisted extraction.

Butterfly pea flower, also known as Asian pigeon wings, has been widely used as an ingredient in food and medicine due to the high content of various types of antioxidants that are beneficial to health. One of the technologies used to extract the bioactive compound from the plant is the ultrasound-assisted extraction (UAE) method. This study aimed to investigate the effect of different extraction times from 10 min to 30 min and types of solvent: water, aqueous methanol and aqueous ethanol on the phenolic compound and antioxidant activity present in the butterfly pea flower. The total phenolic content (TPC) was determined using Folin-Ciocalteu assay while the antioxidant was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay. Based on this study, the optimal condition to increase the extract yield from butterfly pea flower was found to be at a 20-min sonication, while the highest TPC and antioxidant activity were achieved at a 30-min sonication using aqueous methanol. Compared with maceration, UAE demonstrated better results in percentage yield, TPC and antioxidant activity when using aqueous methanol. This approach indicated that UAE is an efficient technique for obtaining bioactive compounds from butterfly pea flowers.

Two techniques, namely, optimized ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE), were used to promote the extraction of phenolic compounds from the pseudo-fruits of Rosa canina L. (RC). For UAE, an optimization process based on the design of experiment (DoE) principles was used for determining the dependence between three variables (i.e., time of extraction, ultrasound amplitude, and the material-to-water ratio) and the total phenolic content of the samples. For EAE, a 2:1:1 pectinase, cellulase, and hemicellulase enzymatic blend was used as pre-treatment for optimized UAE, inducing a higher total phenolic content. The untargeted phenolic profiling approach revealed a great abundance of lower molecular weight phenolics (1.64 mg Eq./g) in UAE-RC extracts, whilst gallic acid (belonging to hydroxybenzoic acid derivatives) was the most abundant individual compound of both extracts. The unsupervised multivariate statistics clearly discriminated the impact of enzymatic pre-treatment on the phenolic profile of RC pseudo-fruits. Finally, Pearson’s correlation coefficients showed that anthocyanins, phenolic acids, and tyrosol derivatives were those compounds mostly correlated to the in vitro antioxidant potential of the extracts, whilst negative and significant (p < 0.05) correlation coefficients were recorded when considering the enzymatic inhibition activities. The highest enzyme-inhibitory activity has been identified against α-glucosidase, which indicates an antidiabetic effect.

The use of by-products as a source of bioactive compounds with economic added value is one of the objectives of a circular economy. The olive oil industry is a source of olive pomace as a by-product. The olive pomace used in the present study was the exhausted olive pomace, which is the by-product generated from the air drying and subsequent hexane extraction of residual oil from the olive pomace. The objective was to extract bioactive compounds remaining in this by-product. Various types of green extraction were used in the present study: solvent extraction (water and hydroalcoholic); ultrasound-assisted extraction; Ultra-Turrax-assisted extraction; and enzyme-assisted extraction (cellulase; viscoenzyme). The phenolic profile of each extract was determined using HPLC-DAD and the total phenolic content (TPC) and antioxidant activity (ABTS, DPPH, and ORAC) were determined as well. The results showed significant differences in the yield of extraction among the different methods used, with the enzyme-assisted, with or without ultrasound, extraction presenting the highest values. The ultrasound-assisted hydroethanolic extraction (USAHE) was the method that resulted in the highest content of the identified phenolic compounds: 2.021 ± 0.29 mg hydroxytyrosol/100 mg extract, 0.987 ± 0.09 mg tyrosol/100 mg extract, and 0.121 ± 0.005 mg catechol/100 mg extract. The conventional extraction with water at 50 °C produced the best results for TPC and antioxidant activity of the extracts. The extracts from the USAHE were able to inhibit Gram-positive bacteria, especially Bacillus cereus, showing 67.2% inhibition at 3% extract concentration.

Apple pomace represents an underexploited source of bioactive compounds. This study examines the optimization of total phenolic content (TPC) and antioxidant extraction yield of apple pomace (variety: Belorusskoje malinovoje) using response surface methodology. The green extraction technique used was ultrasound-assisted extraction, and it was compared with conventional solvent extraction. The impact of extraction time and amplitude of ultrasound-assisted extraction on the yield of polyphenols and antioxidants has been evaluated. Total phenolic content was determined using an established TPC assay. The antioxidant activity of the apple pomace was determined using established assays 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+). Furthermore, the potential of apple pomace as a feed material was explored by assessing its nutritional composition, vitamins, minerals, fatty acids, and amino acid content. The extraction of antioxidants and phenolic compounds was efficiently optimized using RSM. The optimal conditions for TPC and DPPH• analyses were achieved with an extraction time of 17.5 min and an ultrasound-assisted extraction amplitude of 20%. Optimal conditions for ABTS•+ were 5 min extraction time and 20% amplitude. Conventional and ultrasound-assisted extraction methods yielded comparable results. Moreover, apple pomace exhibits potential as a feed ingredient despite its modest protein content. This study contributes to the utilization of apple pomace by providing additional information on its antioxidant content and nutritional composition, thus contributing to its sustainable utilization in various industries, especially the livestock feed sector.

Sacha inchi (Plukenetia volubilis L.) is an oilseed plant that yields a highly nutritious oil. However, its husks and seed shells are under-utilized byproducts. In this study, ethanolic extracts of sacha inchi husks (SI-husk) and seed shells (SI-shell) were obtained using ultrasound-assisted extraction, evaluating the effects of extraction temperature and solvent-to-solid ratio on the yields. The extracts’ total phenolic contents, phenolic profiles, antioxidant activities, and antimicrobial activities against Gram-positive and Gram-negative bacteria were investigated. Under the selected extraction conditions for SI-husk (60 °C, 1:8 s/s, 280W, 120 min) and SI-shell (60 °C, 1:3 s/s, 280W, 120 min), the extracts’ total phenolic contents were 80.18 ± 0.32 and 50.94 ± 0.48 mg GAE (gallic acid equivalents)/g, respectively. Cyanidin, naringenin, and kaempferol were principally found in the SI-husk extract (79.4 %) and vanillic acid in the SI-shell extract (79.9 %). Both extracts exhibited antioxidant activity, with ORAC values of 360.36 ± 0.21 and 228.11 ± 0.14 µmol TE/g, respectively. The antimicrobial activity of the extracts was evaluated against S. aureus, P. aeruginosa, and E. coli using the agar disk diffusion assay. SI-husk (1 mg) exhibited antibacterial activity against Staphylococcus aureus, with an inhibition zone of 10.5 ± 1.8 mm. Our results provide new insights into sacha inchi byproducts as sources of bioactive compounds with potential health benefits.

Saffron is a well-known spice, produced for a long time, mainly in the Mediterranean area. In the last few years, saffron has been seen as an alternative crop for the diversification of agricultural production and a new source of income, due to its high price, including in Alpine areas. Apart from the spice, constituted by the stigmas, saffron production provides a high amount of fresh tepals, which have so far been considered agricultural waste. Recently, studies on the composition of saffron tepals have been revealing their potential as a rich source of bioactive compounds. In this study, we evaluated the productive traits and the quality of the spice of saffron cultivated in open fields in the north western Italian Alps, for 2 years. In addition, in order to valorize the crop residues, we evaluated the bioactive compound content in tepals by using maceration and ultrasound assisted extraction with different rates of water and methanol as solvents. Higher yields in spice were obtained in the second year of cultivation. However, in both years, the spice had a high quality (ISO 3632), and a very high total phenolic content and antioxidant activity. Thirteen bioactive compounds were identified, including flavonols, cinnamic acids, benzoic acids, catechins, and vitamin C, with few differences between sites and year of cultivation. In tepals, the extraction method and the solvent used influenced the evaluated parameters, i.e., total phenolics and total anthocyanins content, antioxidant activity, and the amount of the four phenolic compounds found (hyperoside, rutin, ellagic acid, and epicatechin). Overall, this study revealed that both saffron spice and its by-product, the fresh tepals, produced in the north western Italian Alps can be considered a source of bioactive compounds with nutraceutical properties, having an antioxidant capacity that is often similar or higher than those of some vegetables and fruits.

The increasing interest in healthy and natural foods has raised the attention towards uncommon or unexplored ingredients, such as edible flowers. These products are proven to be a rich source of bioactive compounds, for example, vitamins or polyphenols that play an important role in health promotion and disease prevention. However, plant species with edible flowers are numerous and most of them still need to be studied with this aim. The high species richness of North-Western Italy provides interesting perspectives in the use of wild edible flowers, which are currently underutilized, but can be a valuable food source or food supplement for healthy diets. In this framework, the phytochemical composition of 22 wild edible flowers was analysed and compared with that of four cultivated species (Borago officinalis L., Calendula officinalis L., Tagetes patula L. and Tropaeolum majus L.) to evaluate their potentiality as sources of bioactive compounds. The total polyphenol content (TPC) and antioxidant activity of the fresh flowers were assessed, together with their phenolic profiles and vitamin C content, through spectrophotometric and chromatographic analyses. The evaluated parameters varied widely among species, with Paeonia officinalis L. and Rosa pendulina L. showing the highest values of polyphenols (1,930 mg gallic acid equivalents (GAE) · 100 g−1 and 1,774 mg GAE · 100 g−1, respectively), followed by Rosa canina L. (1,397 mg GAE · 100 g−1) and Geranium sylvaticum L. (1,268 mg GAE · 100 g−1). The same species also showed the highest antioxidant activity, measured with three different assays [ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS)]. The phenolic profile differed among the studied species, with Dianthus pavonius Tausch and R. pendulina having the highest sum of detected polyphenols (2,522 mg · 100 g−1 and 2,366 mg · 100 g−1, respectively). Vitamin C was identified in all but two flowers (Allium ursinum L. and B. officinalis) and Primula veris L. had the highest amount (45 mg · 100 g−1). The study showed that wild edible flowers outperformed the cultivated species, except for T. majus, providing new insights for the use of wild edible flowers as sources of bioactive compounds.

Simple SummaryExhausted olive pomace (EOP) is the main residue of the pomace oil extraction industry, which is generated in large quantities and has limited applications. Thus, this study aimed to obtain bioactive compounds from EOP using ultrasound-assisted extraction as a potential first valorization step. Two types of devices were tested: bath- and probe-type UAE. The operational parameters were studied and optimized to maximize the antioxidant compounds. In particular, hydroxytyrosol was the main phenolic compound identified and its content was 5.16 mg/g EOP (bath-type UAE) and 4.96 mg/g EOP (probe-type UAE). Mannitol was also detected in the extract, 59.53 mg/g EOP (bath-type UAE) and 69.73 mg/g EOP (probe-type UAE). The results highlight the great potential EOP has as a source of bioactive compounds, with applicability in several sectors. Moreover, the probe-type UAE shows potential to be applied for obtaining these bioactive compounds in a continuous and faster manner.Exhausted olive pomace (EOP) is the main agro-industrial waste of the olive pomace extracting industries. It contains phenolic compounds and mannitol, so the extraction of these bioactive compounds should be considered as a first valorization step, especially if EOP is used as biofuel. Therefore, EOP was subjected to bath-type ultrasound-assisted extraction (UAE), and the effects of the acetone concentration (20–80%, v/v), solid load (2–15%, w/v), and extraction time (10–60 min) on the extraction of antioxidant compounds were evaluated according to a Box–Behnken experimental design. By means of the response surface methodology, the optimum conditions were obtained: 40% acetone, 8.6% solids, and 43 min. For all the extracts, the total phenolic content (TPC), flavonoid content (TFC), and antioxidant activity (DPPH, ABTS, and FRAP) were determined. With the aim of shortening the extraction time, a two-level factorial experiment design was also carried out using a probe-type UAE, keeping the solid load at 8.6% (w/v) and the acetone concentration at 40% (v/v), while the amplitude (30–70%) and the extraction time (2–12 min) were varied to maximize the aforementioned parameters. Finally, a maximum of phenolic compounds was reached (45.41 mg GAE/g EOP) at 12 min and 70% amplitude. It was comparable to that value obtained in the ultrasonic bath (42.05 mg GAE/g EOP), but, remarkably, the extraction time was shortened, which translates into lower costs at industrial scale. Moreover, the bioactive compound hydroxytyrosol was found to be the major phenolic compound in the extract, i.e., 5.16 mg/g EOP (bath-type UAE) and 4.96 mg/g EOP (probe-type UAE). Other minor phenolic compounds could be detected by capillary zone electrophoresis and liquid-chromatography–mass spectrometry. The sugar alcohol mannitol, another bioactive compound, was also found in the extract, and its content was determined. Thus, the use of this technology can support the valorization of this waste to obtain bioactive compounds, including mannitol, hydroxytyrosol, and other derivatives, before being applied for other uses.

Laurus nobilis L. is an evergreen Mediterranean shrub whose leaves have been known for various health-promoting effects mainly attributed to polyphenols. Microwave- (MAE) and ultrasound-assisted extraction (UAE) are green extraction techniques that enable effective isolation of polyphenols from plant material. Therefore, the aim of this research was to optimize the extraction conditions of MAE (ethanol percentage, temperature, extraction time, microwave power) and UAE (ethanol percentage, extraction time, amplitude) of polyphenols from Laurus nobilis L. leaves and to assess their polyphenolic profile by ultra performance liquid chromatography- tandem mass spectrometry (UPLC-MS/MS) and antioxidant capacity by oxygen radical absorbance capacity (ORAC) assay. Optimal MAE conditions were 50% ethanol, 80 °C, 10 min and 400 W. Optimal UAE conditions were 70% ethanol, 10 min and 50% amplitude. Spectrophotometric analysis showed the highest total phenolic content in the extracts was obtained by MAE, compared to conventional heat-reflux extraction (CRE) and UAE. The polyphenolic profile of all obtained extracts included 29 compounds, with kaempferol and quercetin glycosides being the most abundant. UPLC-MS/MS showed the highest total phenolic content in the extracts obtained by CRE. ORAC assay showed the highest antioxidant capacity in extracts obtained by CRE, which is in agreement with the polyphenolic profile determined by UPLC-MS/MS.

In this study, the effect of extraction parameters (ethanol concentration, sonication time, and solvent-to-sample ratio) onFicus deltoidealeaves was investigated using ultrasound-assisted extraction by response surface methodology (RSM). Total phenolic content (TPC) ofF. deltoideaextracts was identified using Folin-Ciocalteu method and expressed in gallic acid equivalent (GAE) per g. Box-Behnken statistical design (BBD) was the tool used to find the optimal conditions for maximum TPC. Besides, the extraction yield was measured and stated in percentage. The optimized TPC attained was 455.78 mg GAE/g at 64% ethanol concentration, 10 minutes sonication time, and 20 mL/g solvent-to-sample ratio whereas the greatest extraction yield was 33% with ethanol concentration of 70%, sonication time of 40 minutes, and solvent-to-material ratio at 40 mL/g. The determination coefficient,R2, for TPC indicates that 99.5% capriciousness in the response could be clarified by the ANOVA model and the value of 0.9681 of predictedR2is in equitable agreement with the 0.9890 of adjustedR2. The present study shows that ethanol water as solvent, a short time of 10 minutes, and adequate solvent-to-sample ratio (20 mL/g) are the best conditions for extraction.