Ultrasonic-assisted Extraction Method Research Articles

ULTRASONIC-ASSISTED EXTRACTION OF DIETARY FIBERS FROM FENUGREEK SEEDS

In this study, soluble and insoluble dietary fibers were extracted from fenugreek (Trigonella-foenum graecum L.) seeds using the ultrasonic-assisted extraction method and the fibers were characterized. The effects of the solid–solvent ratio, ultrasonic amplitude and extraction time were investigated using the response surface methodology to obtain the highest total dietary fiber yield. In addition, functional and physicochemical properties of soluble and insoluble dietary fibers such as water holding capacity, oil holding capacity, swelling capacity, glucose adsorption index and ?-amylase inhibition capacity were determined. The optimum process conditions were determined as follows: the solid–solvent ratio of 52.83 g/L, ultrasonic amplitude of 57.86% and extraction time of 24.68 min. At these conditions, insoluble dietary fiber yield was 50.73% and soluble dietary fiber yield was 28.67%. According to the characterization results, insoluble dietary fiber achieved better physicochemical and functional properties compared to soluble dietary fiber.

Extraction, purification and anticancer activity studies on triterpenes from pomegranate peel.

Pomegranate peel is the by-product of pomegranate processing, which contains a lot of triterpene compounds. In this study, the total triterpenes of pomegranate peel (TPP) were extracted using an ultrasonic-assisted ethanol extraction method under optimal conditions, purified using D-101 macroporous resin to obtain a purity of 75.28%. The triterpenes in TPP were mainly pentacyclic triterpenes determined by LC-MS/MS. Network pharmacological analysis predicted that the anticancer targets were closely related to the MAPK pathway. The in vitro results showed that TPP could inhibit cell proliferation, promote apoptosis, reduce mitochondrial membrane potential and increase ROS levels. The western blot results indicated that the expression levels of the apoptotic proteins Bax, Bcl-2, cytochrome C, cleaved caspase-3 and cleaved caspase-9 were increased. In addition, the protein expression of the MAPK pathway predicted by network pharmacology also changed significantly. These results provided that TPP has potential for adjuvant therapy of tumors.

Exploring antioxidant and antidiabetic potential of Mutingia calabura (Kerukupsiam) leaf extract: In vitro analysis and molecular docking study

Antioxidant activity and antihyperglycemic constituents and of traditional medicinal plants are currently the preferred therapeutic means of treatment and management of diabetes because of the undesired adverse effect of synthetic drugs. Muntingia calabura (Kerukupsiam) leaves and other parts are considered as alternative natural sources of treatment for diabetes. Ultrasonic assisted extraction is a novel approach for extraction of phytoconstituents which gives high extraction yield of bioactive compounds. However, there has been no published information presently on the use of ethanol ultrasonic assisted extraction method for assessment of antioxidant and antidiabetic activities of M. calabura leaves. Hence, the current study aims to evaluate the in vitro antioxidant and antidiabetic activities of M. calabura leave extract. IC50 analysis was done to determine theinhibitory concentration and the results obtained from 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay showed IC50 of gallic acid to be 1.0 µg/ml, which is lower than M. calabura leaves extract at 2.54 µg/ml, indicating that only small concentration of gallic acid was required to inhibit the free radicals at 50 %. However, IC50 analysis for amylase inhibition showed that M. calabura extract had 44.39 µg/ml antidiabetic activity compared to acarbose with 57.1 µg/ml activity. This indicates that M. calabura leaves extract has a better inhibition on amylase activity compared to the acarbose which is a synthetic drug. Further still, in silico study was carried out and the molecular docking result of eight ligands against amylase indicates quercetin had the least binding free energy of −9.1 kcal/mol, indicating the strongest interaction. Using Lineweaver-Burk plot, the results showed a competitive inhibition, hence, it was justified that M. calabura has the potential to manage diabetes and other diseases related to free radicals.

Physicochemical, rheological, antioxidant and immunological properties of four novel non-inulin (poly)saccharides from Asparagus cochinchinensis

The impacts of four extraction techniques, including hot water, ultrasonic-assisted, complex enzyme-assisted and acid-assisted methods, on the morphological, physicochemical properties and bioactivities of Asparagus cochinchinensis (poly)saccharides (EACP, WACP, UACP, and AACP) were investigated and compared. The four samples were mainly composed of glucose, fructose, and galactose with molar ratios of 50.8:22.7:4.4 for WACP, 53.9:26.0:5.3 for UACP, 35.6:14.1:21.4 for AACP and 45.0:15.6:9.0 for EACP, respectively. The rheological result showed that ACPs were non-Newtonian fluids. EACP with high purity (97.65 %) had good DPPH, O2− and ABTS+ radical scavenging activities, and significantly promoted the proliferation of the RAW264.7 cells at low concentration. UACP had good Fe2+ chelating ability, radical (DPPH, O2− and OH) scavenging activities, which might be attributed to the existence of triple-helix structure. AACP had high yield, molecular weight (17,477.2 Da), high crystallinity (23.33 %), and good radical (OH and ABTS+) scavenging activities. All four significantly stimulated the transcript expression levels of TNF-α, IL-1β and IL-6, as determined by RT-PCR. These results suggest that the exploitation and utilization of non-inulin (poly)saccharides extracted by ultrasonic-assisted, complex enzyme-assisted and acid-assisted extraction methods are potentially valuable as effective and natural immune adjuvants and antioxidants.

Optimization of Ethanol Extraction Technology for Yujin Powder Using Response Surface Methodology with a Box-Behnken Design Based on Analytic Hierarchy Process-Criteria Importance through Intercriteria Correlation Weight Analysis and Its Safety Evaluation.

Here, we aimed to optimize the ethanol extraction technology for Yujin powder (YJP) and evaluate its safety. The ultrasonic-assisted ethanol reflux extraction method refluxing was used to extract YJP. The parameters were optimized through a combination of single-factor and response surface methodology (RSM). The comprehensive Y value score calculated using the content of 13 active ingredients in YJP ethanolic extracts (YEEs) and the yield of the dry extract were used as measuring criteria. RSM with a Box-Behnken design using three factors and three levels was adopted to optimize the ethanol extraction technology for YJP. Finally, acute and subchronic toxicity tests were performed to evaluate its safety. The results revealed the best technological parameters: a liquid-material ratio of 24:1, an ethanol concentration of 69%, assistance of ultrasound (40 °C, 50 kHZ, 30 min), reflux time of 53 min, and reflux temperature of 50 °C. In acute toxicity tests, the maximum administration dosage in mice was 28.21 g/kg, which is higher than 10 times the clinical dosage. Adverse effects in the acute and subchronic toxicity tests were not observed. All clinical indexes were normal. In conclusion, the RSM based on AHP-CRITIC weight analysis could be used to optimize the ethanol extraction technology for YJP and YEEs prepared under the above conditions and ensure high safety.

Optimization and Kinetics Extraction of Natural Dyes from Henna Leaves (Lawsonia inermis L.) with Ultrasonic Assistance

The Lawson dye contained in the leaves of Lawsonia inermis L. has a red-orange color, which can be used as a fabric dye. The solvent is ethanol because it can attract flavonoid compounds bound to Lawson. The ultrasonic-assisted extraction method is employed to enhance the extraction process by breaking down cell walls, thereby facilitating a more rapid extraction of the desired solute. This study investigated the effects of material-to-solvent ratio, ethanol concentration, and extraction time. The research results were analyzed using a UV-Visible spectrophotometer to determine the yield of Lawson extract. The Face-Centered Composite Design (FCCCD) method was applied to this study to obtain optimal analysis results and extraction conditions. The independent variables in this study were the ratio of ingredients to solvent (0.02-0.06 g/g), ethanol concentration (20-60%), and extraction time (5-15 minutes). The optimal extraction conditions were obtained at a ratio of 0.02 g/g, an ethanol concentration of 60%, and an extraction time of 15 minutes, with a predicted model yield of 15.417%. The actual yield under these conditions was found to be 15.4934%. Furthermore, the extraction kinetics model was analyzed to study and predict the optimal Lawson extraction results. Extraction kinetics calculations were carried out using first-order and second-order based on the Lagergren equation. The most suitable extraction kinetic model is second-order with a determinant coefficient value, or the value of R2 is 1, which indicates that the order kinetic model equation represents the actual conditions.

Ultrasonic-assisted extraction, analysis and identification of water extract of propolis

Apis mellifera is one species of bee that produces propolis, a resin-based product. Propolis extraction using ultrasonic assistance is being widely studied. Using water as a solvent is a challenge to capture the bioactive components of propolis. This research aimed to determine the physicochemical quality resulting from the processing of propolis extract from Central Java by ultrasonics using water as a solvent at different temperatures and times. Raw propolis is extracted by the ultrasonic-assisted extraction method at low, medium, and high temperatures. Raw propolis is extracted by the ultrasonic-assisted extraction method at low, medium, and high temperatures. The study used nine treatments with three replications. The extraction time was carried out for 10, 20, and 30 minutes. The study used nine treatments with three replications. The results of the analysis showed that propolis extraction at different temperatures and times had a very significant effect (P<0.01) on the yield, total phenolic content (TPC), and total flavonoid content (TFC), with an average of 6.7–13.3%, 1.10–2.21 mg GAE/mL, and 0.07–0.32 mg QE/mL, respectively. Propolis extraction at different temperatures and times had no significant effect on tannin content, pH, and antioxidant activity. Regarding yield, TPC, TFC, and tannin content values, it was determined that extracting at high temperatures for 30 minutes produced the best results. High temperatures and long timespans are used for the best chance of collecting bioactive components.

The Effect of Curcumin Nanoparticles on Paracetamol-Induced Liver Injury in Male Wistar Rats.

Curcumin is a naturally occurring compound that has antioxidant properties, acts as a hepatoprotective, and lowers lipid peroxidation. However, curcumin's low solubility and bioavailability are its primary drawbacks and prevent its use as a therapeutic agent. In this study, curcumin nanoparticles will be created using the ultrasonic-assisted extraction method, and their effectiveness against paracetamol-induced changes in ALT, AST, SOD, MDA, and TNF-α will be compared to that of pure curcumin. This study aimed to determine the hepatoprotective effect of curcumin nanoparticles in paracetamol-induced rats as a model for liver injury. Thirty-six male Wistar rats, aged 6 to 8 weeks, with a minimum weight of 120 grams, were used in an experimental laboratory investigation with a post-test-only group design. Rats in each group received 100 mg/kgBW pure curcumin, 100 mg/kgBW curcumin nanoparticles, and 50 mg/kgBW curcumin nanoparticles for 7 days before paracetamol induction. On day 8, 300 mg/kgBW of paracetamol was intraperitoneally injected to cause liver damage. One of the groups received NAC as an antidote 10 hours after paracetamol induction. Detection of ALT and AST using a Chemistry Analyzer. ELISA approach for the detection of SOD, MDA, and TNF-α. The Roenigk score was calculated by two examiners after the liver histopathology preparations were stained using the Hematoxylin-Eosin method. Post hoc analyses were performed after the One Way Annova and Kruskal Wallis tests to examine the data. According to PSA results, the smallest formula that formed curcumin nanoparticles (10.2 nm) was 8 g of curcumin formula mixed with a mixture of Tween 20 4.5 ml, Kolliphor EL 1.5 ml, Propylene Glycol 1.5 ml, and Capryol 90 1 ml for 21 minutes using an ultrasonic process. MDA and TNF-α levels, as well as the liver's histological Roenigk score, were significantly lower in the 100 mg/kgBB pure curcumin group (C100) when compared to the model group (model). The levels of AST, MDA, TNF-α, and the liver histopathology score were significantly lower in the 100 mg/kgBB (NC100) and 50 mg/kgBB (NC50) curcumin nanoparticle groups compared to the model group (model) and pure curcumin group (C100) (p< 0.05). Curcumin nanoparticles showed better hepatoprotective ability than pure curcumin.

Comparison of Different Extraction Methods for Bioactive Compounds from Kao-Kum Doi-Saket (Oryza sativa L.)

Kao-Kum Doi-Saket is one type of purple rice known for its high content of anthocyanin and its ability to resist free radicals. Anthocyanin extracts can be utilized in various applications, including food coloring, nutritional supplements, natural medicine, and cosmetics. There are several methods for extracting anthocyanin from Kao-Kum Doi-Saket. This research aims to compare the effects of extraction methods, including Conventional extraction (CE), Pulse electric field extraction (PEF), and Ultrasonic assisted extraction (UAE) on total anthocyanin content and antioxidant activity in Kao-Kum Doi-Saket. It was found that the UAE method resulted in the highest total anthocyanin content of 57.05 ± 4.27 mg/L, which was significantly different from the other two methods (p&lt;0.05). Kao-Kum Doi-Saket extracts from the UAE method also showed the highest DPPH inhibition (88.32 ± 1.83%) compared to the CE and PEF methods. Therefore, these findings suggest that UAE is the most effective method for extracting anthocyanin and antioxidant activity from Kao-Kum Doi-Saket and could be beneficial for developing processed products from Kao-Kum Doi-Saket in the future.

Ultrasonic-assisted extraction increases the antioxidant and antibacterial capabilities of ciplukan (Physalis angulata) extract

Ciplukan (Physalis angulata) fruit is known to contain various bioactive compounds that can inhibit the growth of pathogenic bacteria and have radical scavenging activity. The purpose of this study is to ascertain how the antioxidant and antibacterial properties of ciplukan extract are affected by ultrasonic-assisted extraction. This study used ciplukan extract that has undergone ultrasonic treatment (P0: 24-hour maceration, P1: 30 minutes, P2: 60 minutes, and P3: 120 minutes). The total phenolic, flavonoid, and antioxidant activity of the ciplukan extract was then assessed using DPPH (2,2′-diphenylpicryl hydrazyl), and the antibacterial activity was assessed using dilution techniques. The outcomes demonstrated that total phenolic contents, flavonoids, and antioxidant activity were significantly impacted by the ultrasonic-assisted extraction method (P<0.05). The highest total phenolic content, flavonoid content, and antioxidant activity (IC50) obtained from the two-hour ultrasonic treatment (P3) were 176.62 mg GAE/g, 121.33mg QE/g, and 50.80 μg/mL, respectively. Furthermore, the capability of ciplukan extract to inhibit the growth of gram-positive and gram-negative bacteria was assessed. The thirty-minute ultrasonic treatment (P1) has the highest antibacterial activity, the MIC values for P1 treatment were 500 μg/mL on Staphylococcus aureus FNCC 0047 and 125 μg/mL on Escherichia coli ATCC 8739 and Pseudomonas aeruginosa ATCC 9027. These results indicate the antioxidant and antibacterial capabilities of ciplukan extract obtained from the physical extraction process using ultrasonic.

Effect of an Amplitude and Time Duration at Betacyanin Extraction by Ultrasonic-Assisted Extraction (UAE) Method

Dragon fruit peel is a byproduct of agriculture that naturally dyes with betacyanin. Application of betacyanin as a natural dye obtained by extraction. In this study, betacyanin was extracted from dragon fruit peel using the Ultrasonic-Assisted Extraction (UAE) technique in order to assess its impact on yield, betacyanin concentration, and redness value. UAE performed extraction with several amplitude levels (60, 72, 84, and 96%) and time durations (4; 10; 16; 22 and 28 minutes). The results significantly affected betacyanin content, redness value, and yield. Based on the Zeleney, the best treatment showed that the extraction with an amplitude of 84% for 16 minutes produced the highest response with betacyanin content, redness value, and yield, respectively of 282.002 mg/L; 48.40 and 64.52%.

Exploring the Biological Functions and Anti-Melanogenesis of Phallus indusiatus for Mushroom-Based Cosmetic Applications

Phallus indusiatus, or bamboo mushroom, has been reported for its nutraceutical properties, while its cosmeceutical properties remain unclear. In this study, we conducted extractions of whole, fresh P. indusiatus using both aqueous and ethanolic methods. Among the extracts, ultrasonic-assisted extraction method with DI showed the highest antioxidant activity compared to the others. For cosmeceutical assessment, we evaluated the extracts’ inhibitory effects against ECM-degrading enzymes and found that they exhibited a modest inhibitory effect of approximately 50%. Remarkably, ultrasonic-assisted extraction with DI demonstrated promising cosmeceutical properties. Additionally, pressure-assisted extraction with DI showed a potentially protective effect against H2O2-induced DNA damage. To investigate the anti-melanogenic effect on MNT-1 cells, we treated them with the extracts and observed a significant decrease in cellular tyrosinase activity under α-MSH stimulation. This resulted in a relative reduction in melanin content. Notably, autoclaving extraction exhibited a significantly greater anti-melanogenic effect than the other extracts at the lowest concentration tested. Furthermore, the extracts demonstrated a reduction in NO production under LPS-induced inflammation. Hot water extraction with DI and ethanol exhibited a stronger anti-inflammatory effect compared to diclofenac, without any cytotoxicity. These findings highlight the hidden cosmeceutical properties of P. indusiatus and suggest its potential use as a bioactive ingredient in cosmetic formulations.

Antibacterial and antioxidant activity of compounds from Citrus sinensis peels and in silico molecular docking study

The increasing prevalence of drug resistance, adverse side effects of existing antibiotics, and the resurgence of previously known infections have necessitated the search for new, safe, and effective antimicrobial agents. The peels of Citrus sinensis (300 g) were extracted using maceration and ultrasonic-assisted extraction methods with ethanol, resulting in yields of 20.99 g and 11.5 g (7%, 7.5%), respectively. Silica gel column chromatographic separation of the ethanol extract yielded N-(1,3,4,5-tetrahydroxydecan-2-y1)octanamide (1), decanoic acid (2), β-sitosterol-3-O-β-D-glucopyranoside derivative (3), and (z)-ethyl tetradec-7-enoate (4). GC-MS analysis of the essential oil detected 7 chemical components accounting for 99.84% of the total composition of which limonene was found to be the predominant constituent (87.5%). In vitro antibacterial tests revealed promising zones of inhibition by ethanol extract (12.67±0.58 mm, at 150 mg/mL), compound 4 (15.67±2.88 mm, at 6 mg/mL), and compound 1 (12.00±0.00 mm, at 6 mg/mL) against E. faecalis, S. typhimurium, and P. aeruginosa, respectively, compared to gentamicin (13.00±1.73 mm, 18.00±1.00 mm, and 16.67±1.15 mm, respectively at 10 µg/mL). DPPH radical scavenging activity indicated that compound 1 exhibited an IC50 value of 0.05 mg/mL, compared to ascorbic acid's 0.016 mg/mL. In silico molecular docking studies revealed that compounds 1 and 3 had the lowest scoring poses against E. coli DNA gyrase B enzyme, human peroxiredoxin 5, and S. aureus pyruvate kinase, respectively. These findings support traditional applications of Citrus peels in treating infectious diseases, particularly against Gram-positive strains, and highlight their potential use as antibacterial ingredients in cosmetics.

Ultrasonic extraction and purification of scutellarin from Erigerontis Herba using deep eutectic solvent

Nowadays, deep eutectic solvent (DES) was widely used in the extraction of bioactive in traditional Chinese medicine (TCM) as an alternative to organic solvents. While as, it is still a challenge for the removal of DES solvents and recovery of the extracted product. In this study, a simple, efficient and green technique of preparing scutellarin from Erigerontis Herba(EH) was proposed, combining ultrasonic assisted-DES extraction with anti-solvent purification. Firstly, different types of DES were prepared and studied for their abilities to extract scutellarin from EH. DES composed of choline chloride and acetamide (1:4 mol/mol) with 30% water obtained the highest extraction yield. Anti-solvent was proved as an efficient method to recover scutellarin from the DES extract with a content of purification up to 71.2%. Moreover, microscopic structural analysis was carried out to investigate the extract process and explain the extraction principle. Furthermore, the antioxidative activities of the DES extracts were evaluated, indicated that the bioactive property of scutellarin were still remained by using DES as the extraction solvent. In conclusion, the proposed simple and green ultrasonic assisted DES extraction method will serve as an effective alternative strategy to extract bioactive compounds from TCM.

Characterization and in vitro antibacterial activity of sulfated polysaccharides from freshwater alga Cladophora crispata.

Barada River is characterized by an abundant growth of freshwater algae. Cladophora sp. algae have emerged as a new source of bioactive compounds. In this research Cladophora crispata was cultivated with the outdoor method, and algal sulfated polysaccharides (SPs) were extracted by an ultrasonic-assisted extraction method. After extraction, gel filtration was used to purify the crude SPs, SP compounds were determined and selected, and the effect of purified SPs as antibacterial agents was investigated. The purified extract gave two fractions (F1 and F2). The chemical components of both crude and purified SPs were then determined. The highest carbohydrate content (74.12%) and protein content (4.02%) was found in the crude extract, while the highest sulfate content (12.17%) was found in purified fraction F2, and the highest uronic acid content (18.46%) was found in purified fraction F1. Fourier transform infrared spectroscopy (FT-IR) was used to confirm that the crude extract and fractions consist of sugar, uronic acids, protein and sulfate groups. Both F1 and F2 consisted of rhamnose, galactose, xylose and ribose based on high performance liquid chromatography (HPLC) separation. Each fraction showed an inhibitory effect on Gram-positive and Gram-negative bacteria. F2 has the lowest minimum inhibitory concentration (MIC) value against Staphylococcus aureus , Bacillus anthracis , Enterobacter aerogenes and Pseudomonas aeruginosa , where its MIC values were 6, 13, 25 and 30 mg ml-1, respectively. Algae polysaccharides are of key interest due to their antibacterial properties, which has led to them being included in pharmaceutics and food applications.

Optimization of ultrasonic-assisted extraction of polyphenol from Areca nut (Areca catechu L.) seeds using response surface methodology and its effects on osteogenic activity

Areca nut (Areca catechu L.) seeds are rich in polyphenols, while few studies focused on it. This study was designed to obtain the maximum extraction yield of areca nut seed polyphenol (ACP). An ultrasonic-assisted extraction method optimized by response surface methodology (RSM) was established to extract ACP. Under the optimal conditions (ultrasonic power of 87 W, ethanol concentration of 65%, extraction temperature of 62℃, and extraction time of 153 min), the actual extraction yield of ACP was 139.62 mg/g. Then we investigated the effects of ACP on the proliferation, differentiation and mineralization of MC3T3-E1 pre-osteoblasts. Results suggested that ACP notably promoted the proliferation of MC3T3-E1 cells without cytotoxicity, and the contents of collagen type Ⅰ (COL-Ⅰ) and osteocalcin (OCN) were rising. Meanwhile, the alkaline phosphatase (ALP) activity and mineralized nodules were enhanced. These findings demonstrated that ACP could induce the proliferation, differentiation and mineralization of osteoblasts in vitro. This work provided a certain experimental basis for the developing and utilization of polyphenols from Areca nut seeds.

Antioxidant Activity of Passiflora edulis (Passion fruit) Seed Extracts Obtained from Maceration and Ultrasonic Assisted Extraction Method

Passion (Passiflora edulis) fruit belongs to the Passifloraceae family and has high-value economics. The extraction efficiency is affected by the phytochemical’s properties of plant, the solvent and extraction method used to extract the desired compounds. The seeds of passion fruit exhibit an oil content, the valuable bioactive compound as antioxidant activity. The passion fruit seed were the waste obtained from the passion fruit syrup company in Makassar City. This study aims to determine the effect of different extraction methods on antioxidant activity of passion fruit seed extracts. The seed was macerated with methanol at room temperature for 5 days, and the ultrasonic-assisted extraction (UAE) process was performed for 30 minutes. The antioxidant capacity was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH). The results showed that the percentage yield of extract from UAE was 6.45% and the extract yield from maceration method was 5.458%. The result of antioxidant activity (IC50 value) revealed that the methanol extract obtained from UAE and maceration were 71.672 ug/mL and 144.901 ug/mL, respectively. These results indicate that the UAE method is a suitable extraction method to produce strong antioxidant activity from Passiflora seed extract.

Betulinic acid, a major therapeutic triterpene of Celastrus orbiculatus Thunb., acts as a chemosensitizer of gemcitabine by promoting Chk1 degradation.

Celastrus orbiculatus Thunb., also called as oriental bittersweet vine or climbing spindle berry, a traditional Chinese herbal medicine has been used to treat a spectrum of painful and inflammatory diseases for centuries. Explored for their unique medicinal properties, C.orbiculatus offers additional therapeutic effects on cancerous diseases. The effect of single-agent gemcitabine on survival has not long been encouraging, combination therapies provide patients multiple chances of benefit for improved clinical response. This study aims at expounding the chemopotentiating effects and underlying mechanisms of betulinic acid, a primary therapeutic triterpene of C. orbiculatus in combination with gemcitabine chemotherapy. The preparation of betulinic acid was optimized using ultrasonic-assisted extraction method. Gemcitabine-resistant cell model was established by induction of the cytidine deaminase. MTT, colony formation, EdU incorporation and Annexin V/PI staining assays were used to evaluate cytotoxicity, cell proliferation and apoptosis in BxPC-3 pancreatic cancer cell line and H1299 non-small cell lung carcinoma cell line. Comet assay, metaphase chromosome spread and γH2AX immunostaining were applied for DNA damage assessment. Western blot and co-immunoprecipitation was used to detect the phosphorylation and ubiquitination of Chk1. Mode of action of gemcitabine in combination with betulinic acid was further captured in BxPC-3-derived mouse xenograft model. We noticed that the extraction method had an impact on the thermal stability of C. orbiculatus. Ultrasound-assisted extraction at room temperature in shorter processing time could maximize the overall yields and biological activities of C. orbiculatus. The major constituent was identified as betulinic acid, and the pentacyclic triterpene represented the prominent anticancer activity of C. orbiculatus. Forced expression of cytidine deaminase conferred acquired resistance to gemcitabine, while betulinic acid displayed equivalent cytotoxicity toward gemcitabine-resistant and sensitive cells. A combination therapy of gemcitabine with betulinic acid produced synergistic pharmacologic interaction on cell viability, apoptosis and DNA double-strand breaks. Moreover, betulinic acid abrogated gemcitabine-triggered Chk1 activation by destabilizing Chk1 loading via proteasomal degradation. The combination of gemcitabine and betulinic acid significantly retarded BxPC-3 tumor growth in vivo compared to single-agent gemcitabine treatment alone, accompanied with reduced Chk1 expression. These data provide evidence that betulinic acid is a potential candidate for chemosensitization as a naturally occurring Chk1 inhibitor and warrants further preclinical evaluation.

Effects of ultrasonic assisted high-temperature cooking method on the physicochemical structure characteristics and in vitro antioxidant capacities of dietary fiber from Dendrocalamus brandisii Munro shoots

In this study, the ultrasonic assisted high-temperature cooking extraction method of soluble dietary fiber from bamboo shoots was optimized by response surface methodology, and the effects of ultrasonic assisted high-temperature cooking extraction on the structural characteristics, physicochemical properties and antioxidant activity of soluble dietary fiber (SDF) from bamboo shoots were evaluated. The yield of modified UH-SDF11UH-SDF: Extraction of soluble dietary fiber from Dendrocalamus brandisii Munro shoots by ultrasonic assisted high-temperature cooking method. was significantly higher than that of untreated D-SDF22D-SDF: Untreated extracted soluble dietary fiber from Dendrocalamus brandisii Munro shoots.. FTIR and XRD confirmed that UH-SDF had more hydrophilic groups and higher crystallinity (28.73 %), resulting in better thermal stability. SEM observation showed that UH-SDF exhibited a more loose microstructure, and the particle size of UH-SDF (601.52 μm) was significantly smaller than that of D-SDF (242.59 μm), so UH-SDF had a larger specific surface area. In addition, UH-SDF has stronger water holding capacity, water swelling capacity and oil holding capacity than D-SDF. The DPPH radical and hydroxyl radical scavenging rates of UH-SDF were 8.91 % and 7.49 % higher than those of D-SDF. In addition, the reducing ability of UH-SDF was higher than that of D-SDF, which had better antioxidant activity. In summary, UH-SDF has the potential to be developed as an anti-inflammatory functional food.

Anthocyanins and Total Phenolic Compounds from Pigment Extractions of Non-Native Species from the Umia River Basin: Eucalyptus globulus, Tradescantia fluminensis, and Arundo donax

Alien invasive species (AIS) and non-native species are a prominent and extended problem in a wide range of areas in Europe and around the world. Centered in the Umia’s riparian forest, in Galicia, we found at least three main AIS needing to be controlled and harvested to preserve the biodiversity of the area. Previous studies probed that leaves and bark of selected species—Tradescantia fluminensis, Arundo donax, and Eucalyptus globulus—have important antioxidant properties, suitable for use in pharmaceutical and industrial contexts. A comparison of four solid-liquid extraction methods—Soxhlet extraction, ultrasound assisted extraction, thermal agitator, and infusion—was conducted in order to define the most efficient method in correlation within antioxidant (anthocyanins and total phenols) extraction. Water was selected as solvent, providing a sustainable research background without implying any chemical additives. The best extraction yields were obtained with Soxhlet extraction for all raw matter, with best results for Tradescantia fluminensis (41.89%) and Eucalyptus globulus leaves (39.35%); followed by the ultrasonic assisted extraction method, with better yield performed with Eucalyptus leaves (27.07%). On the contrary, Tradescantia fluminensis showed better efficiency with thermal agitator (35.07% compared to 23.19% from ultrasound extraction). After extractions, identification and quantification of total phenolic compounds and anthocyanins were carried out using spectrophotometric determination and acid hydrolysis in butanol method. In general, the best extraction yield in correlation with higher antioxidant content was performed by thermal agitator method, and Eucalyptus globulus leaves were demonstrated to be the better anthocyanins (6.18 ± 0.82 mg CC/g) and total phenols containers (172.40 ± 44.53 mg GAE/g). Studies provided promising possibilities for the residues of the three non-native species analyzed, as a source of antioxidants, favoring circular economy systems, as well as taking care of biodiversity in affected environments.