Extraction Of Total Flavonoids Research Articles

Identification of Key Hypolipidemic Components and Exploration of the Potential Mechanism of Total Flavonoids from Rosa sterilis Based on Network Pharmacology, Molecular Docking, and Zebrafish Experiment.

Hyperlipidemia is a prevalent chronic metabolic disease that severely affects human health. Currently, commonly used clinical therapeutic drugs are prone to drug dependence and toxic side effects. Dietary intervention for treating chronic metabolic diseases has received widespread attention. Rosa sterilis is a characteristic fruit tree in China whose fruits are rich in flavonoids, which have been shown to have a therapeutic effect on hyperlipidemia; however, their exact molecular mechanism of action remains unclear. Therefore, this study aimed to investigate the therapeutic effects of R. sterilis total flavonoid extract (RS) on hyperlipidemia and its possible mechanisms. A hyperlipidemic zebrafish model was established using egg yolk powder and then treated with RS to observe changes in the integral optical density in the tail vessels. Network pharmacology and molecular docking were used to investigate the potential mechanism of action of RS for the treatment of hyperlipidemia. The results showed that RS exhibited favorable hypolipidemic effects on zebrafish in the concentration range of 3.0-30.0 μg/mL in a dose-dependent manner. Topological and molecular docking analyses identified HSP90AA1, PPARA, and MMP9 as key targets for hypolipidemic effects, which were exerted mainly through lipolytic regulation of adipocytes and lipids; pathway analysis revealed enrichment in atherosclerosis, chemical carcinogenic-receptor activation pathways in cancers, and proteoglycans in prostate cancer and other cancers. Mover, chinensinaphthol possessed higher content and better target binding ability, which suggested that chinensinaphthol might be an important component of RS with hypolipidemic active function. These findings provide a direction for further research on RS interventions for the treatment of hyperlipidemia.

Green and efficient extraction of flavonoids from Perilla frutescens (L.) Britt. leaves based on natural deep eutectic solvents: Process optimization, component identification, and biological activity

In this study, an ultrasonic-assisted natural deep eutectic solvent (NaDES) was used to extract flavonoids from Perilla frutescens (L.) Britt. leaves. Of 10 tested NaDESs, that comprising D-(+)-glucose and glycerol exhibited the best total flavonoid extraction rate. Response surface methodology (RSM) was used for extraction modeling and optimization, and the total flavonoid content reached 87.48 ± 1.61 mg RE/g DW, which was a significant increase of 5.36% compared with that of 80% ethanol extraction. Morphological changes in P. frutescens leaves before and after extraction were analyzed by scanning electron microscopy (SEM), and the mechanism of NaDES formation was studied by Fourier transform infrared (FT-IR) spectroscopy. Furthermore, 10 flavonoids were identified by UPLC-Q-TOF-MS. In addition, the NaDES extract had better biological activity according to five kinds of antioxidant capacity measurements, cyclooxygenase-2 (COX-2) and hyaluronidase (Hyal) inhibition experiments. Moreover, the stability test revealed that the total flavonoid loss rate of the NaDES extract after four weeks was 37.75% lower than that of the ethanol extract. These results indicate that the NaDES can effectively extract flavonoids from P. frutescens leaves and provide a reference for further applications in the food, medicine, health product and cosmetic industries.

Assessment of Antioxidant Activity, Total Phenolic and Flavonoid Contents of Albizia saponaria L. Bark Extract

Langir (Albizia saponaria L.), belonging to the family of the Fabaceae, is a medicinal endemic plant of South Sulawesi, Indonesia. The Albizia genus shows antioxidant, antibacterial, anthelmintic, antidandruff, and anti-inflammatory properties. In this study, total phenolic (TP) and total flavonoid (TF) extracts from the bark of A. saponaria were screened for robust antioxidant activity in order to identify potential sources of new compounds for use in pharmaceutical formulations in the future. TP and TF of the 96% ethanol extract and fractions (hexane, ethyl acetate, butanol, and water) were calculated by the Folin–Ciocalteu and aluminum chloride procedures, respectively. Furthermore, the antioxidant activity was determined by DPPH free radical scavenging method and ABTS assay. Generally, both extract and fractions showed significant radical scavenging activities. Ethyl acetate fraction exhibited more potent radical scavenging activity in the DPPH method (IC50 35.27 ± 2.85 µg/mL) and ABTS assay (IC50 60.04 ± 0.98 µg/mL), followed by 96% ethanol extract, and hexane, butanol, and water fractions. Furthermore, the highest TP (4.50 ± 0.01 mg/g GAE) and TF (3.55 ± 0.04 mg/g QE) were obtained from ethyl acetate fraction. There was a strong correlation between antioxidant activity with TP (DPPH, R2 = 0.6436; ABTS, R2 = 0.7676) and TF content (DPPH, R2 = 0.5533; ABTS, R2 = 0.5961). The extract’s antioxidant properties may be attributable to its higher phenolic and flavonoid content. In summary, the phenolic and flavonoid content of the ethyl acetate fraction indicates its potential utility as a source of antioxidants.

Effects of Rhizopus-arrhizus-31-Assisted Pretreatment on the Extraction and Bioactivity of Total Flavonoids from Hibiscus manihot L.

The Hibiscus manihot L. (HML) Medic, an edible hibiscus of the Malvaceae family, is abundant with flavonoids. The study investigated how Rhizopus-arrhizus-31-assisted pretreatment affects the extraction and bioactivity of flavonoids from HML. The fiber structure of the fermented flavonoid sample (RFF) appears looser, more porous, and more disordered than the unfermented flavonoid sample (RUF). RFF demonstrates milder conditions and yields higher extraction rates. According to the Box-Behnken response surface optimization experiment, the optimal conditions for RFF include a material-liquid ratio of 1:41 g/mL, a 2 h extraction time, a 57% ethanol concentration, and an extraction temperature of 800 °C, resulting in a 3.69% extraction yield, which is 39.25% higher than that of RUF. Additionally, RFF exhibits greater activity than RUF in the radical-scavenging system. The IC50 values for DPPH, OH, and ABTS radicals are 83.43 μg/mL and 82.62 μg/mL, 208.38 μg/mL and 175.99 μg/mL, and 108.59 μg/mL and 75.39 μg/mL for RUF and RFF, respectively. UPLC-QTOF-MS analysis of the active components in the HML flavonoid sample revealed significant differences in the chromatograms of RUF and RFF, indicating that biofermentation led to substantial changes in composition and content from HML.

Optimization of Microwave-Assisted Extraction Process of Total Flavonoids from Salicornia bigelovii Torr. and Its Hepatoprotective Effect on Alcoholic Liver Injury Mice.

The objective of this study was to determine the optimal extraction conditions for total flavonoids from S. bigelovii using microwave-assisted extraction and to analyze the protective effect of total flavonoids from S. bigelovii on alcoholic liver injury in mice. The optimization of the process conditions for the microwave-assisted extraction of total flavonoids from S. bigelovii was performed using response surface methodology, and an alcohol-induced acute liver injury model in mice was used to investigate the effects of different doses of total flavonoids (100 mg/kg, 200 mg/kg, and 400 mg/kg) on the levels and activities of serum alanine aminotransferase kits (ALT), glutamic oxaloacetic transaminase kits (AST), superoxide dismutase kits (SOD), glutathione peroxidase kits (GSH-Px), and malondialdehyde (MDA). We performed hematoxylin-eosin (H&E) staining analysis on pathological sections of mouse liver tissue, and qRT-PCR technology was used to detect the expression levels of the inflammatory factors IL-1 β, IL-6, and TNF-α. The results revealed that the optimal extraction process conditions for total flavonoids in S. bigelovii were a material-to-liquid ratio of 1:30 (g/mL), an ethanol concentration of 60%, an extraction temperature of 50 °C, an ultrasound power of 250 W, and a yield of 5.71 ± 0.28 mg/g. Previous studies have demonstrated that the flavonoids of S. bigelovii can significantly inhibit the levels of ALT and AST in the serum (p < 0.001), reduce MDA levels (p < 0.001), increase the activity of the antioxidant enzymes SOD and GSH-Px (p < 0.001), and inhibit the IL-1 β, IL-6, and TNF-α gene expression levels (p < 0.001) of inflammatory factors. The total flavonoids of S. bigelovii exert a protective effect against alcoholic liver injury by reducing the levels of inflammation, oxidative stress, and lipid peroxidation caused by alcohol. The results of this study lay the foundation for the high-value utilization of S. bigelovii and provide new resources for the development of liver-protective drugs.

Enzyme-assisted ultrasonic extraction of total flavonoids and extraction polysaccharides in residue from Abelmoschus manihot (L)

Abelmoschus manihot (L) is a traditional chinese herb and the present study focused on its comprehensive development and utilization. Enzyme-assisted ultrasonic extraction (EUAE) was investigated for the extraction and qualitative and quantitative analysis of flavonoids from Abelmoschus manihot (L) using a combination of ultra-performance liquid chromatography-photodiode array (UPLC-PDA), polysaccharides was extracted from residues and compared with directly extracted from raw materials. The optimal yield of 3.46±0.012 % (w/w) was obtained when the weight ratio of cellulase to pectinase was 1:1, the enzyme concentration was 3 %, the pH was 6.0, the solvent was a mixture of 70 % ethanol (v/v) and 0.1 mol/L NaH2PO4 buffer solution, the ultrasonic power was 500 W, the extraction time was 40 min, and the temperature of the extraction was 50 °C. The individual concentrations of interested flavonoids (rutin, neochlorogenic acid, nochlorogenic acid, lsoquercitrin, quercitrin, gossypin, quercetin) were effectively increased with the using of EUAE, compared with ultrasonic extraction (UE) method. Polysaccharides were extracted from each residue, respectively, the Polysaccharides yield in residue from EUAE was higher than that from UE, and closed to the yield from direct extraction in raw materials. The above results shown that the experimental process had the potential to be environmentall, friendly, straightforward and efficient.

Optimization of the Flavonoid Extraction Process from the Stem and Leaves of Epimedium Brevicornum and Its Effects on Cyclophosphamide-Induced Renal Injury.

Cyclophosphamide (CTX) is a broad-spectrum alkylated antitumor drug. It is clinically used in the treatment of a variety of cancers, and renal toxicity is one of the adverse reactions after long-term or repeated use, which not only limits the therapeutic effect of CTX, but also increases the probability of kidney lesions. The total flavonoids of Epimedium stem and leaf (EBF) and Icariin (ICA) are the main medicinal components of Epimedium, and ICA is one of the main active substances in EBF. Modern pharmacological studies have shown that EBF has a variety of biological activities such as improving osteoporosis, promoting cell proliferation, antioxidant and anti-inflammatory properties, etc. However, few studies have been conducted on the nephrotoxicity caused by optimized CTX extraction, and protein-ligand binding has not been involved. This research, through the response surface optimization extraction of EBF, obtained the best extraction conditions: ethanol concentration was 60%, solid-liquid ratio of 25:1, ultrasonic time was about 25 min. Combined with mass spectrometry (MS) analysis, EBF contained ICA, ichopidin A, ichopidin B, ichopidin C, and other components. In this study, we adopted a computational chemistry method called molecular docking, and the results show that Icariin was well bound to the antioxidant target proteins KEAP1 and NRF2, and the anti-inflammatory target proteins COX-2 and NF-κB, with free binding energies of -9.8 kcal/mol, -11.0 kcal/mol, -10.0 kcal/mol, and -8.1 kcal/mol, respectively. To study the protective effect of EBF on the nephrotoxicity of CTX, 40 male Kunming mice (weight 18 ± 22) were injected with CTX (80 mg/kg) for 7 days to establish the nephrotoxicity model and were treated with EBF (50 mg/kg, 100 mg/kg) for 8 days by gavage. After CTX administration, MDA, BUN, Cre, and IL-6 levels in serum increased, MDA increased in kidney, GPT/ALT and IL-6 increased in liver, and IL-6 increased in spleen and was significant ((p < 0.05 or (p < 0.01)). Histopathological observation showed that renal cortex glomerular atrophy necrosis, medullary inflammatory cell infiltration, and other lesions. After administration of EBF, CTX-induced increase in serum level of related indexes was reduced, and MDA in kidney, GPT/ALT and IL-6 in liver, and IL-6 in spleen were increased. At the same time, histopathological findings showed that the necrosis of medullary and corticorenal tubular epithelium was relieved at EBF (50 mg/kg) dose compared with the CTX group, and the glomerular tubular necrosis gradually became normal at EBF (100 mg/kg) dose. Western blot analysis of Keap1 and Nrf2 protein expression in kidney tissue showed that compared with model CTX group, the drug administration group could alleviate the high expression of Keap1 protein and low expression of Nrf2 protein in kidney tissue. Conclusion: After the optimal extraction of total flavonoids from the stems and leaves of Epimedium, the molecular docking technique combined with animal experiments suggested that the effective component of the total flavonoids of Epimedium might activate the Keap1-Nrf2 signaling pathway after treatment to reduce the inflammation and oxidative stress of kidney tissue, so as to reduce kidney damage and improve kidney function. Therefore, EBF may become a new natural protective agent for CTX chemotherapy in the future.

Choline chloride-urea-based natural deep eutectic solvent for highly efficient extraction of polyphenolic antioxidants from Pluchea indica (L.) Less leaves

Pluchea indica leaves (PIL) is a polyphenolic-rich plant. Polyphenols, such as phenolics and flavonoids, are natural antioxidants. This study aims to evaluate the ability of natural deep eutectic solvent (NADES) based on ultrasonic-assisted extraction (UAE) for highly efficient extraction of the polyphenolic antioxidant compounds from PIL. Choline chloride-urea (ChCl-U) was selected based on screening of 17 types of NADES. This study uses Response Surface Methodology (RSM) to help determine optimal extraction conditions. The selected variables include molar ratio, temperature, and the water addition in the NADES. The responses used for the RSM study were total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and reducing power (RP) methods with microtiter analysis. The best condition for polyphenolic antioxidants extraction using ChCl-U NADES was a 1:3 molar ratio, temperature of 52 °C, and 26% of water addition in NADES. Under these conditions, the extraction of total phenolics and flavonoids from PIL with antioxidant activity increased up to 1.2 folds and 3.1 folds compared with 50%-ethanol and methanol, respectively. The morphology of the leaves before and after extraction was also evaluated with a scanning electron microscope (SEM) to observe the effects of ultrasonic vibrations with different solvents. Furthermore, the compound di-caffeoylquinic acid (DCQA) in PIL was selectively found in ChCl-U extract through quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS/MS) analysis. Thus, ChCl-U combined with UAE can be developed for polyphenolic extraction from PIL for extract production as an antioxidant.

Guadua angustifolia Kunth leaves as a source for bioactive phenolic compounds: Optimization of ultrasound-assisted extraction using response surface methodology and antioxidant activities

This study evaluated the ultrasound-assisted extraction of phenolic compounds from Guadua angustifolia leaves, along with their optimization using response surface methodology. The effects of two sonication process conditions were determined using a central composite experimental design, with three levels (low, medium, and high) evaluated for time (10, 20, and 30 min) and temperature (20 °C, 35 °C, and 50 °C). A total of 12 experiments with four replicates were conducted at the central point, with the total phenol and flavonoid contents determined using the Folin-Ciocalteu colorimetric method and complexation with AlCl3, respectively. The optimized extract was analyzed using ultra-performance liquid chromatography (UPLC), and the antioxidant capacity of the optimized extract was determined by DPPH• (2,2-Diphenyl-1-Picrylhydrazyl) and ABTS•+ (2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) assays. Extraction at 50 °C for 20 min was found to favor the extraction of phenol and total flavonoids. The experimental validation of the total phenol and flavonoid content produced values of 7.39 mg gallic acid equivalents per gram of dry matter and 1.55 mg quercetin equivalents per gram of dry matter, respectively. These values suggest that the extraction process is reproducible, with a relative standard deviation of 22.9 % and 14.1 %, respectively. The chromatographic profile showed that optimization favored the visualization of phenolic compounds compared to the non-optimized extract. The optimized extract had higher antioxidant capacity than the non-optimized extract, with values of 209.23 and 144.76 μmol Trolox per gram extract for the DPPH• and ABTS•+ techniques, respectively. Thus, the conditions evaluated in the ultrasound-assisted extraction were an efficient technique capable of extracting the maximum amount of phenolic compounds with antioxidant activity from the leaves of G. angustifolia, showing its potential application in various industries.

Comparative efficacy of ultrasound-assisted and hot water extraction of papaya leaves

IntroductionThis article describes the optimal process-product characteristics of a bath type sonication and hot water system based aqueous extraction of Carica papaya leaves. MethodsUltrasound assisted extraction of papaya leaves was conducted to assess the optimal sonication time, extraction temperature and loading ratio with respect to the combinatorial extraction of optimal total polyphenols, total flavonoids and anti-oxidant activity. The experimental runs were designed with the face centred design module of the Design Expert software (Version 7.0.0) and involved 20 alternate combinations (in triplicate) of the chosen degrees of freedom and mentioned responses. Further, hot water extraction was chosen as a control with design variable values set as the optimal value data point obtained from the response surface based numerical optimisation. ResultsDuring both pulse and normal ultrasound extraction of papaya leaves, it was found that the loading ratio followed by extraction temperature and sonication time had a hierarchal critical influence on all evaluated responses. Numerical optimisation subsequently confirmed higher loading ratio at moderate combination of extraction temperature and sonication time. Compared to the reported data, higher response values were obtained in this work. These have been attributed to the sensitive optimality of process parameters and the choice of shade dried leafy samples. ConclusionThis study showed the promising scope of ultrasound-assisted extraction based papaya leafy extraction process and its complimentary role in fostering the utility of unconventional leafy systems-based aqueous extracts in commercial dry herbal supplements.

Flavonoid extract from propolis alleviates periodontitis by boosting periodontium regeneration and inflammation resolution via regulating TLR4/MyD88/NF-κB and RANK/NF-κB pathway

Ethnopharmacological relevanceIn traditional Chinese medicine, propolis has been used for treating oral diseases for centuries, widely. Flavonoid extract is the main active ingredient in propolis, which has attracted extensive attention in recent years. Aim of the studyThe objective and novelty of the current study aims to identify the mechanism of total flavonoid extract of propolis (TFP) for the treatment of periodontitis, and evaluate the therapeutic effect of TFP-loaded liquid crystal hydrogel (TFP-LLC) in rats with periodontitis. MethodsIn this study, we used lipopolysaccharide-stimulated periodontal ligament stem cells (PDLSCs) to construct in vitro inflammation model, and investigated the anti-inflammatory effect of TFP by expression levels of inflammatory factors. Osteogenic differentiation was assessed using alkaline phosphatase activity and alizarin red staining. Meanwhile, the expression of toll like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), nuclear factor-kappa B (NF-κB), receptor activator of NF-κB (RANK) etc, were quantitated to investigate the therapeutic mechanism of TFP. Finally, we constructed TFP-LLC using a self-emulsification method and administered it to rats with periodontitis via periodontal pocket injection to evaluate the therapeutic effects. The therapeutic index, microcomputed tomography (Micro-CT), H&E staining, TRAP staining, and Masson staining were used for this evaluation. ResultsTFP reduced the expression of TLR4, MyD88, NF-κB and inflammatory factor in lipopolysaccharide-stimulated PDLSCs. Meanwhile, TFP simultaneously regulating alkaline phosphatase, RANK, runt-associated transcription factor-2 and matrix metalloproteinase production to accelerate osteogenic differentiation and collagen secretion. In addition, TFP-LLC can stably anchor to the periodontal lesion site and sustainably release TFP. After four weeks of treatment with TFP-LLC, we observed a decrease in the levels of NF-κB and interleukin-1β (IL-1β) in the periodontal tissues of rats, as well as a significant reduction in inflammation in HE staining. Similarly, Micro CT results showed that TFP-LLC could significantly inhibit alveolar bone resorption, increase bone mineral density (BMD) and reduce trabecular bone space (Tb.Sp) in rats with periodontitis. ConclusionCollectively, we have firstly verified the therapeutic effects and mechanisms of TFP in PDLSCs for periodontitis treatment. Our results indicate that TFP perform anti-inflammatory and tissue repair activities through TLR4/MyD88/NF-κB and RANK/NF-κB pathways in PDLSCs. Meanwhile, for the first time, we employed LLC delivery system to load TFP for periodontitis treatment. The results showed that TFP-LLC could be effectively retained in the periodontal pocket and exerted a crucial role in inflammation resolution and periodontal tissue regeneration.

Green synthesis and biophysical characterization of silver and palladium nanoparticles using Laureliopsis philippiana: A potent eco-friendly larvicide with negligible impact on zebrafish (Danio rerio)

Green nanoparticles represent a sustainable and eco-friendly approach to developing larvicides targeted at controlling mosquito populations. These nanoparticles are synthesized using natural, non-toxic materials and environmentally benign processes, emphasizing their potential to combat mosquito larvae while minimizing adverse effects on the ecosystem. This approach aligns with the growing need for sustainable solutions in vector control, providing an effective alternative to traditional larvicides with reduced environmental impact. The aim of the present investigation we have utilized the leaf from Laureliopsis philippiana (LP) to synthesize silver (Ag) and palladium (Pd) nanoparticles (NPs) and studied toxicity on deadly disease vectors larvae of Aedes aegypti and Anopheles stephensi. Biosynthesized Ag and Pd NPs were extensively characterized through physical and chemical methods. Gas chromatography–mass spectrometry analysis of L. philippiana assisted in identifying the secondary metabolites of the extract's total flavonoid and phenolic compounds, which are in the reactants of the bio-reduction process during the production of NPs. The in-silico analysis was conducted on mosquito sterol carrier protein-2 (1PZ4) and mosquito odorant binding protein (2L2C). The biosynthesized Ag and Pd NPs revealed the UV absorption peak at 417 and 370 nm, and X-ray diffraction results showed the crystalline structure of NPs. At the same time, in transmission electron microscopy representation, the Ag and Pd NPs were spherical, clustered, and exhibited a noticeable average size of Ag NPs: 13.25 ± 2.53 nm and Pd NPs: 4.07 ± 1.64 nm. The biogenic Ag and Pd NPs showed concentration-dependent larvicidal mortality at 12 and 24-hour exposure with LC50 values of 29.596, 17.804, and 23.915, 14.878 µg/mL on Ae. aegypti and 23.946, 13.938, 19.457, and 12.876 µg/mL against An. stephensi, correspondingly. In-silico studies proved that Laureliopsine A and isoterpinolene showed significant docking scores compared to permethrin on mosquito proteins (1PZ4 and 2L2C). Besides, the LP-Ag and Pd NPs non-toxicity analysis was carried out on zebrafish (Danio rerio) embryos at 72 h post-fertilization. Overall, the findings clearly demonstrated the green synthesized Ag and Pd NPs from L. philippiana to develop environmentally secure and safe larvicides.

A green approach for multivariate consecutive extraction of essential oils and flavonoids from Citrus aurantium L. var. amara Engl.: Process optimization and mechanistic insights based on machine learning methods

In this study, a green approach for the multivariate consecutive extraction of essential oils and total flavonoids from Daidaihua was proposed, which combined classical steam distillation with alkali extraction and acid precipitation. The process parameters were optimized through single-factor experiments and Box-Behnken design, and the model for evaluating and predicting the process was constructed by combining artificial neural network and genetic algorithm. The artificial neural network and genetic algorithm model demonstrated satisfactory predictive capabilities, and the following optimal parameters resulted: solid-liquid ratio 1:15.3 (g:mL), distillation time 94.1 h, NaOH concentration 5.4%, extraction temperature 46.7 ℃and extraction time 52.9 min. Under these conditions, the extraction yields of essential oils and total flavonoids reached 0.81 ± 0.07% and 5.52 ± 0.18%, respectively. Significantly, the mass transfer mechanism of the multivariate consecutive extraction process was clearly elucidated using the mass transfer theory model and physical characterization. The essential oils were vaporized from the Daidaihua cells along with water vapor, promoting the destruction of cell structure, reducing the extraction resistance of flavonoids, and speeding up the diffusion of sodium hydroxide solution rapidly into the cell interior and dissolving the flavonoids, improving the efficiency of the mass transfer of flavonoids from the inside to the outside of the cell, consequently the flavonoids extraction yield and efficiency were significantly improved. Obviously, the extraction process dovetailed neatly with environmentally friendly and low-carbon needs. Meanwhile, the composition of the essential oils and total flavonoids was identified by GC-MS and HPLC-MS analysis, respectively. Overall, this research presented a green and clean multivariate consecutive extraction process of Daidaihua, which could contribute to promoting the upgrading and sustainable development of the plant extraction industry.

Comparative analysis of flavonoids extracted from Dendrobium chrysotoxum flowers by supercritical fluid extraction and ultrasonic cold extraction

Flowers are by-products of the medicinal plant Dendrobium chrysotoxum, with high yield but low utilization rate. Extracting flavonoids from D. chrysotoxum flowers can not only reduce resource waste but also bring economic benefits. In this study, two green extraction methods, SFE (supercritical fluid extraction) and UCE (ultrasonic cold extraction) were used to extract flavonoids from D. chrysotoxum flowers, and the response surface method (RSM) was used to optimize the extraction conditions. The results showed that on the optimal conditions of SFE (time 90 min, temperature 50 °C, pressure 20 MPa) and UCE (time 30 min, power 400 W, temperature 40 °C, solid-liquid ratio 1:40 (w:v), ethanol concentration 60%), the total flavonoid extraction yield was 2.04% ± 0.02% and 0.64% ± 0.02%, respectively. In vitro, antioxidant activity analysis showed that both extracts had good antioxidant activity, and the antioxidant activity of SFE extract was higher than that of UCE extract. The quantitative determination of flavonoid compounds showed that the content of quercetin, kaempferol, and catechin in SFE extract was significantly higher than that in UCE extract, and the content of ellagic acid in the two extracts was similar. These results indicated that the two green extraction methods improved the resource utilization of D. chrysotoxum flowers, and SFE was more suitable for extracting flavonoids from D. chrysotoxum flowers than UCE.

Optimization of ultrasonic-assisted extraction and purification of total flavonoids with biological activities from Radix Puerariae

Optimization of ultrasonic-assisted extraction and purification of total flavonoids with biological activities from Radix Puerariae

Green Waste from Cucumber (Cucumis sativus L.) Cultivation as a Source of Bioactive Flavonoids with Hypolipidemic Potential

Cucumber is an important agricultural crop that is cultivated under greenhouse conditions. Cucumber cultivation generates substantial quantities of green waste that require proper disposal. The lack of data on the chemical composition of cucumber greens and their biological activity emphasizes the necessity for research on useful products resulting from this widely available waste. Our examination of the flavonoid contents in the leaves and stems of 30 cucumber cultivars revealed 6 cultivars with high flavonoid contents. In addition, the cutting time did not cause negative effects, and we observed a positive effect with 0.05–0.10% organomineral fertilizer application on the flavonoid levels in the greens. Liquid chromatography–mass spectrometry detected 38 apigenin derivatives, including acylated and non-acylated cucumerins and C-, O-, and C,O-glycosides. Among these, 12 known flavonoids and 18 novel compounds were identified. The concentrations of these compounds in the six flavonoid extracts varied at 39.85–181.53 mg/g for the non-acylated flavones, 14.67–293.31 mg/g for the cucumerins, and 401.73–892.17 mg/g for the acylated flavones. Oral administration of the total flavonoid extracts (at a dosage of 100 g/kg/day) resulted in a hypolipidemic effect in hyperlipidemic hamsters, with subsequent normalization of their serum lipid profiles, malondialdehyde levels, and liver antioxidative enzyme activities. These results substantiate the lipid-lowering potential of cucumber waste extracts. Our investigation of the selected flavonoid activity showed that isovitexin-2′′-O-glucoside-6′′-O-p-coumarate (administered at 50 g/kg/day) had the highest hypolipidemic potency. These results can contribute to the practical use of cucumber green waste and the development of novel supplements for diseases linked to high-fat consumption.

Quantitative determination of total flavonoids in Glycyrrhiza Glabra L. herbs

Licorice herb (Glycyrrhiza glabra L.) is a promising herbal raw material, which can be comprehensively used to develop drugs with an anti-inflammatory action.The aim of the article was to development a quantitative determination method of total flavonoids in Glycyrrhiza glabra L. herbs.Materials and methods. The subjects of research were 5 samples of licorice herb harvested in summer in various places of growing and cultivation. Pinostrobin was used as a standard sample. The registration of the electronic spectra was carried out with a spectrophotometer (Analytik Jena AG, Germany) by differential spectrophotometry, 96% ethanol was used as a solvent.Results. The methods for quantitative determination of total flavonoids in Glycyrrhiza glabra L. was carried out at an analytical wavelength of 310 nm equivalent to pinocembrin. The optimum parameters for the extraction of total flavonoids from Glycyrrhiza glabra L. were as follows: the extractant – 90% ethanol; the «raw material-extractant» ratio was 1:50; the extraction time was 60 min; the degree of atomization was 2 mm. The content of total flavonoids for the Glyccyrhiza glabra L. herb has been determined, it varies from 0.39±0.002 to 3.41±0.015% with the humidity of the vegetative raw material from 9.97±0.003 to 10.03±0.003% depending on the place of the vegetation, cultivation and year of the raw material collection. The error of the single determination with a 95% confidence level was ±0.73%.Conclusion. The developed methods for the quantitative determination of total flavonoids in Glycyrrhiza glabra L. herbs can be used to solve the issues of standardization of these medicinal plant raw materials.

Extraction technology, components analysis and anti-inflammatory activity in vitro of total flavonoids extract from Artemisia anomala S. Moore

Artemisia anomala S. Moore exerts many pharmacological activities, including the removing of the blood stasis, relieving of the fever and analgesia, reducing the swelling and dampness. In this study, the extraction technology, chemical compositions and anti-inflammatory effect in vitro and mechanism of total flavonoids extract from Artemisia anomala S. Moore were studied. The optimal yield rate of total flavonoids extract was optimized by single factor experiments and response surface method, and the chemical constituents were analyzed by UPLC-QTOF-MS method; and the anti-inflammatory activity of the extract was evaluated with lipopolysaccharide induced RAW 264.7 cells. The highest extraction rate was 2.02% under these conditions of the concentration of ethanol 50%, the ultrasonic extraction time 30 min, and the ratio of solvent volume to material weight 20:1 (ml/g). In addition, the main components of total flavonoid extract were preliminarily identified and deduced based on mass spectrometry information and relevant literatures, and its stronger anti-inflammatory activity was demonstrated by reducing the phagocytosis, the content of nitric oxide and the level of related cytokines (tumor necrosis factor-α, interleukin−10, interleukin-6). Furthermore, it was further revealed that the anti-inflammatory effect of the extract was closely connected with the activation of TLR4-MyD88-NF-κB signalling pathway. This study indicated that the total flavonoids extract from Artemisia anomala S. Moore may be a better candidate anti-inflammatory natural medicine.

Extraction optimization and constituent analysis of total flavonoid from Hosta plantaginea (Lam.) Aschers flowers and its ameliorative effect on chronic prostatitis via inhibition of multiple inflammatory pathways in rats

Ethnopharmacological relevanceHosta plantaginea (Lam.) Aschers flowers (HPF) are well-known for their high flavonoid content, which contribute to their widely as traditional Chinese medicine for alleviating inflammation. Despite their recognized potential, information regarding the total flavonoid (TF) of HPF and its therapeutic application in treating chronic prostatitis (CP) remains unknown. Aim of the studyWe aimed to investigate the extraction optimization, constituent analysis, and alleviating effect of TF on CP as well as its potential mechanism. Materials and methodsThe optimized extraction of TF from HPF was explored using response surface methodology with a Box-Behnken design model. The major flavonoids in TF were identified based on UHPLC-MS approach. Efficacy of TF (25 and 100 mg/kg, p.o.) on CP was evaluated in prostate antigen emulsion-induced autoimmune CP rat model by measuring prostatic index, the levels of leukocytes and lecithin bodies, as well as histopathological examination. The protein expression contents were detected by western blotting. Additionally, the antioxidant (DPPH and ABTS) and anti-inflammatory (cyclooxygenase 2, COX-2 inhibitory) effects of TF were also evaluated in vitro. ResultsThe optimized conditions for TF extraction were determined as 60% ethanol concentration, 30 mL/g liquid-to-solid ratio, 30 min extraction time, and 90 °C extraction temperature, and the extraction ratio is 65.98 ± 2.14%. A total of 15 major flavonoids in TF were characterized by comparison with reference standards. TF ameliorated the efficacy of CP in rats in a dose-independent manner, including reduced prostatic index and leukocytes levels, elevated lecithin body levels, ameliorated histopathological damage to prostate, and suppressed phosphorylated protein expressions of nuclear factor kappa-B (NF-κB) p65, inhibitor of NF-κB alpha (IκBα), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (Erk), just another kinase 1 (JAK1), signal transducer and activator of transcription 3 (STAT3), phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt). Simultaneously, the IC50 of TF to DPPH, ABTS radicals, and COX-2 were 2.02, 1.79, and 0.0838 mg/mL, respectively. ConclusionsWe first demonstrated that TF from HPF represents a promising candidate to alleviate CP through suppression of NF-κB, MAPKs, JAK-STAT, and PI3K-Akt signaling pathways.

A network pharmacology approach to decipher the total flavonoid extract of Dracocephalum Moldavica L. in the treatment of cerebral ischemia- reperfusion injury.

Cerebral ischemia-reperfusion injury (CIRI) is a major injury that seriously endangers human health and is characterized by high mortality and high disability. The total flavonoid extract of Dracocephalum moldavica L.(TFDM) in the treatment of CIRI has been proved by clinical practice. But the mechanism for the treatment of CIRI by TFDM has not been systematically revealed. The active compounds contained in TFDM were screened by literature mining and pharmacokinetic parameters, and the targets related to CIRI were collected by searching Drugbank, Genecards and OMIM databases. Cytoscape software was used to construct the protein interaction network of TFDM for the prevention and treatment of CIRI. Geneontology and signal pathway enrichment were analyzed. The key target pathway network of TFDM compounds was constructed and verified by pharmacological experiment in vitro. 21 active components were screened, 158 potential drug targets for the prevention and treatment of CIRI were obtained, 53 main targets were further screened in the protein-protein interaction network, and 106 signal pathways, 76 biological processes, 26 cell components and 50 molecular functions were enriched (P<0.05). Through the compound-target-pathway network, the key compounds that play a role in the prevention and treatment of CIRI, such as acacetin, apigenin and other flavonoids, as well as the corresponding key targets and key signal pathways, such as AKT1, SRC and EGFR were obtained. TFDM significantly decreased LDH, MDA levels and increased the NO activity levels in CIRI. Further studies have shown that TFDM increases the number of SRC proteins, and TFDM also increases p-AKT/ AKT. Molecular docking results showed that acacetin-7-O (- 6''-acetyl) -glucopyranoside, acacetin7-O-β-D-glucopyranoside, apigenin-7-O-β-D-galactoside respectively had good affinity for SRC protein. Acacetin-7-O (- 6''-acetyl) -glucopyranoside,acacetin-7-O-β-D-glucuronide, acacetin7-O-β-D-glucopyranoside had good affinity for AKT1 protein, respectively. Our research showed that TFDM had the characteristics of multi-component, multi-target and multi-channel in the treatment of CIRI. The potential mechanism may be associated with the following signaling pathways:1) the signaling pathways of VEGF/SRC, which promote angiogenesis, 2) the signaling pathways of PI3K/AKT, which inhibit apoptosis, and 3) acacetin-7-O (- 6''-acetyl) -glucopyranoside is expected to be used as a candidate monomer component for natural drugs for further development.