Green and Efficient Ultrasonic-Assisted Extraction of Bioactive Components from Salvia miltiorrhiza by Natural Deep Eutectic Solvents.
ABSTRACT: Natural deep eutectic solvents (NaDESs) are recently developed green solvent alternatives to conventional fossil solvents. The present work systematically screened 22 different NaDESs for the ultrasonic-assisted extraction of bioactive components from Salvia miltiorrhiza (SM), a widely used traditional Chinese medical plant. The suitable solvent and extraction condition were optimized in a two-round screening. In comparison with fossil solvents, NaDESs, especially L-proline-lactic acid (L-Pro-Lac) showed significant advantages in the extraction of salvianolic acid B (SAB), tanshinone IIA (TIIA) and cryptotanshinone (CYT). The optimized yields of the three targeting compounds were 42.05, 1.485 and 0.839 mg/g, respectively. The present method was also applied to the pretreatment of SM samples from different geographic origins. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of NaDES extracts were determined in the study to prove the feasibility of NaDES in bioactive component extraction. The application of NaDESs in the extraction of both hydrophilic and hydrophobic small molecules from SM is proved to be a green and efficient method for pretreatment of herbal materials.
Project description:<i>Chamaenerion angustifolium</i> (L.) Scop. (fireweed) is a perennial herbaceous plant of the Onagraceae family widely used in folk and scientific medicine. It is a promising source of bioactive components. One of the modern trends in extraction is the use of natural deep eutectic solvents (NADESs) combined with ultrasound-assisted extraction (UAE). However, works devoted to the extraction of biologically active substances from <i>C. angustifolium</i> using NADESs are scarce. The aim of this work is a comprehensive study of UAE of bioactive components from <i>C. angustifolium</i> using NADESs based on choline chloride and malonic, malic, tartaric, and citric acids. The antioxidative properties, total phenols, and flavonoids content were estimated for NADES-based extracts. The reference solvents were water and 90% <i>v</i>/<i>v</i> ethanol. Volatile extracted components were identified using GC-MS. The kinetics of the UAE were studied at 45 °C for 20-180 min with water added to 30 wt% NADES. The power of the ultrasound was 120 W, and the frequency was 40 kHz. It was found that NADES choline chloride + citric acid is more effective for the extraction of bioactive components. For this, NADES UAE conditions were optimized following a Box-Behnken design of the experiment and a response surface methodology. The temperature ranged from 30 to 60 °C, the time of extraction ranged from 20 to 60, and the addition of water ranged from 30 to 70 wt%. We established the optimal extraction conditions: temperature 58 °C, time of extraction 35 min, and 70 wt% water. The obtained results expand the knowledge about the use of NADES for the extraction of biologically active compounds from cheap and available plant raw materials.
Project description:Natural deep eutectic solvent (NaDES) is generally considered as a greener alternative to fossil solvent, with great potential in various areas. In the present work, 25 different NaDESs were screened for the extraction of puerarin (PUE) and its two natural derivatives from <i>Radix Pueraria</i> (RP). As the main isoflavone in RP, PUE has a wide range of biological activities. However, its application is restricted due to its poor solubility in water and low oral bioavailability. In this study, the extraction of PUE with NaDESs showed significant advantages compared with traditional solvents. While using L-Pro-Maa (L-proline-malic acid) under optimal conditions, the optimized yields of PUE, 3-MPR and PRX were 98.7 mg/g, 16.3 mg/g and 9.9 mg/g, respectively, which were 2.2-, 2.9- and 3.4-fold higher than that of water. Furthermore, the oral bioavailability of PUE in NaDES extracts was comparatively investigated in rats with HPLC-MS technique. Pharmacokinetic analysis revealed that the relative bioavailability of PUE in L-Pro-Maa extract is 323%. The result indicated that NaDES is not only a sustainable ionic liquid with higher extraction efficiency, but also an enhancer of oral bioavailability of specific natural products.
Project description:Aiming to fulfil the sustainability criteria of future biorefineries, a novel biomass pretreatment combining natural deep eutectic solvents (NaDESs) and microwave (MW) technology was developed. Results showed that NaDESs have a high potential as green solvents for lignin fractionation/recovery and sugar release in the following enzymatic hydrolysis. A new class of lignin derived NaDESs (LigDESs) was also investigated, showing promising effects in wheat straw delignification. MW irradiation enabled a fast pretreatment under mild condition (120 °C, 30 min). To better understand the interaction of MW with these green solvents, the dielectric properties of NaDESs were investigated. Furthermore, a NaDES using the lignin recovered from biomass pretreatment as hydrogen bond donor was prepared, thus paving the way for a "closed-loop" biorefinery process.
Project description:In recent years, natural deep eutectic solvents (NADESs) have gained increasing attention as promising nontoxic solvents for biotechnological applications, due to their compatibility with enzymes and ability to enhance their activity. Betaine-based NADESs at a concentration of 25 wt % in a buffered aqueous solution were used as media to inhibit thermal inactivation of POXA1b laccase and its five variants when incubated at 70 and 90 °C. All the tested laccases showed higher residual activity when incubated in NADES solutions, with a further enhancement achieved also for the most thermostable variant. Furthermore, the residual activity of laccases in the presence of NADESs showed a clear advantage over the use of NADESs' individual components. Molecular docking simulations were performed to understand the role of NADESs in the stabilization of laccases toward thermal inactivation, evaluating the interaction between each enzyme and NADESs' individual components. A correlation within the binding energies between laccases and NADES components and the stabilization of the enzymes was demonstrated. These findings establish the possibility of preincubating enzymes in NADESs as a facile and cost-effective solution to inhibit thermal inactivation of enzymes when exposed to high temperatures. This computer-aided approach can assist the tailoring of NADES composition for every enzyme of interest.
Project description:Primary plant metabolites can be used for artificial preparation of natural deep eutectic solvents (NADESs), which have strong dissolving capacity, good biocompatibility, and biodegradability. In this study, for the first time, we verified that NADESs were present in <i>Coptidis Rhizoma</i> extract and systematically investigated its effects and mechanisms on the pharmacokinetics of oral berberine hydrochloride (BBR), a co-existing bioactive constituent. First, three LC-MS/MS based methods were established and fully validated to determine the levels of 11 primary metabolites in <i>Coptidis Rhizoma</i> extract. According to the weight ratio of four major primary metabolites in the <i>Coptidis Rhizoma</i> extract, a stable "endogenous" NADES was prepared using the heating method by the addition of 350 <i>μ</i>l of water to 1,307.8 mg of the mixture of malic acid (490.5 mg), glucose (280.6 mg), sucrose (517.7 mg), and choline chloride (19.0 mg). The prepared NADES showed significant acute toxicity in mice and cytotoxicity in MDCK-MDR1 cells. However, after being diluted 10 times or 100 times, the NADES had no significant acute toxicity or cytotoxicity, respectively. The dilutions of the NADES significantly increased the water solubility of BBR, reduced its efflux in gut sacs and MDCK-MDR1 cell monolayer, and improved its metabolic stability in intestinal S9. In addition, the NADES dilutions reversibly opened the tight junctions between the enterocytes in the gut sacs. Moreover, the NADES dilutions significantly improved the exposure levels of BBR in the portal vein and livers of mice that were administered oral BBR. Malic acid was identified as a major component in the NADES in terms of solubility, acute toxicity, cytotoxicity, and pharmacokinetic-improving effects on oral BBR. In conclusion, the primary metabolites of <i>Coptidis Rhizoma</i> extract could form "endogenous" NADES, and its dilutions improve the pharmacokinetics of oral BBR. This study demonstrates the synergistic interaction of the constituents of <i>Coptidis Rhizoma</i> extract and the potential use of the NADES dilutions in oral BBR delivery.
Project description:Peumus boldus Mol., is a Chilean medicinal tree used for gastrointestinal and liver diseases. Such medicinal properties are associated with the presence of bioactive flavonoids and aporphine alkaloids. In this study, a new green and efficient extraction method used seven natural deep eutectic solvents (NADES) as extraction media. The extraction efficiency of these NADES was assessed, determining the contents of boldine and total phenolic compounds (TPC). Chemical profiling of P. boldus was done by high-performance liquid chromatography coupled to photo diode array detector and electrospray ion-trap mass spectrometry (HPLC-PDA-ESI-IT/MS) and electrospray ionization quadrupole time-of-flight high-resolution mass spectrometry (HPLC-ESI-QTOF-MS). Among the NADES tested, NADES4 (choline chloride-lactic acid) and NADES6 (proline-oxalic acid) enable better extraction of boldine with 0.427 ± 0.018 and 2.362 ± 0.055 mg of boldine g-1 of plant, respectively. Extraction of boldine with NADES4 and NADES6 was more efficient than extractions performed with methanol and water. On the other hand, the highest TPC were obtained using NADES6, 179.442 ± 3.79 mg of gallic acid equivalents (GAE g-1). Moreover, TPC in extracts obtained with methanol does not show significant differences with NADES6. The HPLC-PAD-MS/MS analysis enable the tentative identification of 9 alkaloids and 22 phenolic compounds. The results of this study demonstrate that NADES are a promising green extraction media to extract P. boldus bioactive compounds and could be a valuable alternative to classic organic solvents.
Project description:The present study aimed at evaluating the effectiveness of different natural deep eutectic solvents (NADES) on the extraction of phenolic compounds from <i>Lavandula pedunculata</i> subsp. <i>lusitanica</i> (Chaytor) Franco, on the antioxidant activity, and acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and tyrosinase (Tyr) inhibitory capacities. Ten different NADES were used in this research and compared with conventional solvents. Ultrasound-assisted extraction (UAE) for 60 min proved to be the best extraction condition, and proline:lactic acid (1:1) and choline chloride:urea (1:2) extracts showed the highest total phenolic contents (56.00 ± 0.77 mg<sub>GAE</sub>/g<sub>dw</sub>) and antioxidant activity [64.35 ± 1.74 mg<sub>TE</sub>/g<sub>dw</sub> and 72.13 ± 0.97 mg<sub>TE</sub>/g<sub>dw</sub> in 2.2-diphenyl-1-picrylhydrazyl (DPPH) and 2.2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods, respectively]. These extracts also exhibited enzymes inhibitory capacity particularly against Tyr and AChE. Even so, organic acid-based NADES showed to be the best extractants producing extracts with considerable ability to inhibit enzymes. Twenty-four phenolic compounds were identified by HPLC-HRMS, being rosmarinic acid, ferulic acid and salvianolic acid B the major compounds. The results confirmed that the combination of UAE and NADES provide an excellent alternative to organic solvents for sustainable and green extraction, and have huge potential for use in industrial applications involving the extraction of bioactive compounds from plants.
Project description:A novel, efficient extraction procedure based on natural deep eutectic solvents (NADES) and ionic liquids (ILs) for determination of 20-hydroxyecdysone (20-E) in spinach has been developed. NADES, the first green extraction agent, with different hydrogen bond donors and acceptors are screened in order to determine extraction efficiencies. NADES consisting of lactic acid and levulinic acid at a molar ratio of 1:1 exhibits the highest yields. ILs, the second green extraction agent, with various cations and anions are also investigated, where [TEA] [OAc]·AcOH, ?AcOH = 0.75 displays the highest recovery. Moreover, NADES-SLE and IL-SLE (SLE, solid-liquid extraction) parameters are investigated. Using the obtained optimized method, the recoveries of the target compound in spinach are above 93% and 88% for NADES-SLE and IL-SLE procedure, respectively. The methods display good linearity within the range of 0.5-30 ?g/g and LODs of 0.17 µg/g. The proposed NADES-SLE-UHPLC-UV and IL-SLE-UHPLC-UV procedures can be applied to the analysis of 20-E in real spinach samples, making it a potentially promising technique for food matrix. The main advantage of this study is the superior efficiency of the new, green extraction solvents, which results in a significant reduction of extraction time and solvents as compared to those in the literature.
Project description:Extracts rich in bioactive compounds added to edible films have allowed the development of active packaging that increases the shelf life of food. However, it is necessary to search for solvents that are nontoxic and not harmful to the environment, with natural deep eutectic solvents (NADES) being an attractive and easily synthesized alternative. This research aimed to design NADES by lyophilization to be used in the extraction of anthocyanins from the Chilean <i>Luma chequen</i> (Molina) A. Gray berry, and subsequently adding them to the matrix of edible ?-carrageenan films. For this purpose, ultrasound-assisted extraction (UAE) was used and the anthocyanin content was evaluated with the pH differential method. The antioxidant capacity of extracts was determined by DPPH assay and the antibacterial capacity by diffusion agar tests. The results obtained indicate that the designed NADES are efficient at extracting anthocyanins, reaching concentrations between 81.1 and 327.6 mg eq cyanidin 3-glucoside/100 g dw of <i>L. chequen</i> (Molina) A. Gray. The extracts reached inhibition diameters between 5 and 34 mm against <i>Escherichia coli</i>, <i>Staphylococcus aureus</i>, and <i>Salmonella typhi</i> strains. Once the extracts were incorporated into ?-carrageenan films, active edible films with antioxidant and antibacterial capacities were obtained.
Project description:<h4>Abstract</h4> Olive leaf as an agricultural waste contains valuable bioactive compounds that are mainly used for pharmaceutical and cosmetic industries. Lately the major component, oleuropein, has gained extra attention due to the anti-viral activity against SARS-CoV-2 that causes Coronavirus disease (Covid-19). In this study, extraction of the bioactive compounds from olive leaves was conducted using a non-conventional and green method. New generation green solvents, natural deep eutectic solvents (NADES) were used in combination with ultrasound assisted extraction. Screening of NADES type, temperature, and particle size were investigated using one-pot-at-a-time method while, NADES amount and liquid-to-solid ratio were optimized using experimental design. The results were evaluated in terms of total polyphenol yield (YTP), total flavonoid yield (YTF) and antiradical activity (AAR). At the optimized conditions, the highest total polyphenol yield and the highest total flavonoid yield were achieved with choline chloride–fructose–water (CFW) (5:2:5) as 187.31?±?10.3 mg gallic acid equivalent g?1 dw and 12.75?±?0.6 mg apigenin equivalent g?1 dw, respectively. The extracts were also analyzed for oleuropein, caffeic acid and luteolin contents. The highest amount of oleuropein and caffeic acid were extracted by glucose–fructose–water (GFW) (1:1:11) as 1630.80 mg kg?1 dw and 112.77 mg kg?1 dw, respectively. <h4>Graphic Abstract</h4> <h4>Supplementary Information</h4> The online version of this article (10.1007/s12649-021-01411-3) contains supplementary material, which is available to authorized users.