The inhibitory activity of Flos Sophorae Immaturus extract and its major flavonoid components on pancreatic lipase.

Inhibition of pancreatic lipase (PL) is a strategy to prevent obesity. The inhibitory effects of Flos Sophorae Immaturus (FSI) extract and its main flavonoid components, rutin and quercetin, on PL were investigated. The contents of rutin and quercetin in FSI extract were 44.10 ± 1.33 % and 6.07 ± 1.62 %, respectively. The IC50 values of FSI extract, rutin and quercetin on PL were 322, 258 and 71 µg/mL, respectively. Rutin and quercetin inhibited PL in a reversible and noncompetitive manner. The combination of rutin and quercetin exhibited synergistic inhibitory effects at low concentration. The binding of rutin/quercetin with PL caused the fluorescence quenching of protein. Fluorescence titration showed the binding affinity of quercetin with PL protein was stronger than that of rutin. Circular dichroism analysis showed the binding changed the secondary structure of PL with an increase in random coil and a decrease in α-Helix and ß-Sheet. Molecular docking revealed that rutin and quercetin could interact with the amino acid residues around the catalytic site through multiple secondary interactions. In vivo studies showed that FSI extract can reduce fat absorption and promote fecal fat excretion through inhibition of PL activity, and the effects were mainly due to rutin and quercetin.

Biosynthesized Silver Nanoparticles Inhibit Osteoclastogenesis by Suppressing NF-κB Signaling Pathways.

Osteoclasts overactivity plays a critical role in the progress of inflammatory bone loss. In addition, ROS can facilitate the formation and function of osteoclasts. Silver nanoparticles (Ag NPs) with ROS scavenging activity are potential candidates for inflammatory bone loss. In this regard, the biosynthetic Ag NPs with low toxicity and high stability by using Flos Sophorae Immaturus extract as the reducing and capping agents are reported. The inflammatory bone loss model is established by injecting LPS. Quantitative reverse transcription-polymerase chain reaction and Western Blot are utilized to determine the expression level of target biomarkers related to osteoclast formation. Ag NPs can significantly reduce the number of TRAP-positive (TRAP+ ) cells. In addition, Ag NPs down-regulate the expression of biomarkers relevant to osteoclast formation. Interestingly, Ag NPs can effectively suppress osteoclast formation via down-regulating ROS-mediated phosphorylation of NF-κB pathways. The in vivo study shows that Ag NPs can ameliorate bone density and decrease osteoclast number. Due to these benefits, the constructed Ag NPs can delay the progression of inflammatory bone loss. These findings suggest that Ag NPs are a potential therapeutic agent in the treatment of inflammatory bone loss.

A new strategy for browning regulation: Flos Sophorae Immaturus extract and thermal treatment modulates nitric oxide and reactive oxygen species network in fresh-cut potatoes.

Browning discoloration is a critical issue that negatively affects the quality of fresh-cut products and their industrial growth. Although many individual anti-browning technologies have been adopted, very few reports on the combination use of natural product extracts and physical methods exist. This study investigated the use of Flos Sophorae Immaturus extract in conjunction with thermal treatment and discovered that the combination effectively retarded browning in fresh-cut potatoes. Accordingly, the activities of polyphenol oxidase, peroxidase, and phenylalanine ammonia-lyase, as well as phenol accumulation, were effectively regulated. Meanwhile, this combination treatment also allowed for the modulation of nitric oxide synthase, superoxide dismutase, and catalase activities, while also regulating the concentrations of nitric oxide, superoxide anion, and hydrogen peroxide. Furthermore, the duplex treatment also regulated the antioxidant capacity and malondialdehyde concentrations. In addition, 39 phytoactive compounds, including protocatechuic acid, quercetin, (-)-alpha-pinene, and matrine, were identified in the extract, which may function as the anti-browning composition. These findings suggest that the combination technology modulated the dynamic equilibrium of production and clearance of nitric oxide and reactive oxygen species, thereby reducing browning deterioration. This is, to our knowledge, the first report of the combined application of Flos Sophorae Immaturus and thermal treatment, which may offer a novel option for fresh-cut preservation. PRACTICAL APPLICATION: The feasibility of integrating a novel highly efficient, safe, environmentally friendly, and easy-to-operate anti-browning alternative, with the ability to integrate into the existing processing line was investigated. The color of sliced potato chips was significantly improved (73.4%) by dipping them in a 0.01% Flos Sophorae Immaturus solution for 5 min and then in 55°C water for 2 min. In this regard, superior browning alleviation may depend on the regulation of the browning reaction and the NO-ROS network. This method has a promising future for making fresh-cut products more appealing to consumers and may provide guidance for fresh-cut producers and related industries.

In Vitro Inhibitory Effects of Polyphenols from Flos sophorae immaturus on α-Glucosidase: Action Mechanism, Isothermal Titration Calorimetry and Molecular Docking Analysis.

Flos sophorae immaturus (FSI) is considered to be a natural hypoglycemic product with the potential for a-glucosidase inhibitory activity. In this work, the polyphenols with α-glucosidase inhibition in FSI were identified, and then their potential mechanisms were investigated by omission assay, interaction, type of inhibition, fluorescence spectroscopy, circular dichroism, isothermal titration calorimetry and molecular docking analysis. The results showed that five polyphenols, namely rutin, quercetin, hyperoside, quercitrin and kaempferol, were identified as a-glucosidase inhibitors with IC50 values of 57, 0.21, 12.77, 25.37 and 0.55 mg/mL, respectively. Quercetin plays a considerable a-glucosidase inhibition role in FSI. Furthermore, the combination of quercetin with kaempferol generated a subadditive effect, and the combination of quercetin with rutin, hyperoside and quercitrin exhibited an interference effect. The results of inhibition kinetics, fluorescence spectroscopy, isothermal titration calorimetry and molecular docking analysis showed that the five polyphenols were mixed inhibitors and significantly burst the fluorescence intensity of α-glucosidase. Moreover, the isothermal titration calorimetry and molecular docking analysis showed that the binding to α-glucosidase was a spontaneous heat-trapping process, with hydrophobic interactions and hydrogen bonding being the key drivers. In general, rutin, quercetin, hyperoside, quercitrin and kaempferol in FSI are potential α-glucosidase inhibitors.

Influence of Ultrasound-Assisted Vacuum Drying on Physicochemical Characteristics, Antioxidant Activity, and α-Glucosidase Inhibition Activity of Flos Sophorae Immaturus.

Flos Sophorae Immaturus (FSI) contains a large number of bioactive substances with antioxidant and hypoglycaemic activity. However, a feasible drying process plays an important role in the retention of its biological activity. The present work investigated the effects of ultrasound-assisted vacuum drying (UAVD) on FSI samples in terms of drying time, colour, microstructure, and total flavonoid content (TFC). Meanwhile, the antioxidant activity and α-glucosidase inhibition activity were also evaluated. The results show that the drying time of UVAD samples was decreased by 40% compared to that of the single vacuum-dried (VD) samples (600 W for 10 min). The cellular porous structures of FSI tissue were formed by UAVD, which promoted the migration of water from the inside to the outside. Furthermore, samples treated by UAVD exhibited better antioxidant activities and α-glucosidase and α-amylase inhibition capacities, with DPPH (81.86%), ABTS (88.61%), FRAP (83.05%), α-glucosidase inhibition capacity (89%), α-amylase (85%), drying time (3 h), and total aberration (ΔE) (1.63) being the highest characteristic traits. In this condition, the highest levels of total flavonoid content (TFC), rutin, quercetin, kaempferol, isorhamnetin, and genistein were obtained with 266.94, 239.46, 35.56, 8.54, 10.37, and 5.64 mg/g DW, respectively. The results confirm that UAVD is a novel method that significantly reduced the VD time and promoted the release of the bioactive substances of FSI.

Hydrothermal treatment improves xanthine oxidase inhibitory activity and affects the polyphenol profile of Flos Sophorae Immaturus.

BACKGROUND: Flos Sophorae Immaturus (FSI) is rich in polyphenols and a potential uric acid-lowering food. However, the processing of FSI is greatly restricted due to the heat sensitivity and low solubility of polyphenols. In this study, hydrothermal treatment - an effective strategy - was applied to FSI processing. The variation of xanthine oxidase (XO) inhibitory effect and polyphenol composition of FSI during hydrothermal treatment were recorded. RESULTS: The XO inhibition rate of FSI increased from 32.42% to 89.00% after hydrothermal treatment at 220 °C for 30 min, as well as total polyphenols (from 0.66 to 1.11 mg mL-1 ) and flavonoids (from 1.21 to 1.58 mg mL-1 ). However, high thermal temperature (>160 °C) and extended thermal time (>90 min) caused the degradation of polyphenols. Rutin, kaempferol-3-O-rutinoside and narcissoside rapidly degraded and converted to quercetin, kaempferol and isorhamnetin when the temperature exceeded 160 °C. The maximum yields of quercetin, kaempferol and isorhamnetin were at 220 °C for 30 min, 90 min and 90 min, respectively. Meanwhile, the conversion kinetics conformed to the first-order model. Interestingly, these newly formed polyphenols possessed better XO inhibitory effects than their derivatives with 3-O-rutinoside. CONCLUSION: Polyphenol conversion during hydrothermal treatment was the main reason for enhancing XO inhibitory activity. Therefore, hydrothermal treatment is an appropriate method for improving the XO inhibitory effect of FSI. © 2022 Society of Chemical Industry.

The prebiotic properties of polysaccharides obtained by differentiated deproteinization methods from Flos Sophorae Immaturus on Lactobacillus fermentum.

The polysaccharides derived from various deproteinization methods were prepared from Flos Sophorae Immaturus (FSI) to investigate the prebiotic efficacy of Lactobacillus fermentum (L.f ). The implications of polysaccharides from FSI (PFSI) gained after purification performed by non-deproteinization and different deproteinization processes (Savage method, papain method, and TCA method) via one-factor optimization were firstly investigated for the influences on the growth of L.f. The utilization of carbohydrate sources and the synthesis of protein and lactate during its growth were analyzed, as well as the variations of LDH, SOD, and GSH- Px enzyme dynamics. The results showed that the one-factor optimization of the deproteinization process with the protein removal rate and polysaccharide retention rate as the indexes led to the optimal methods of the Sevage method with 5 elution times, papain method with 80 U/mL concentration, and TCA method with 2.5 ratio, respectively. In addition, the PFSI obtained with or without deproteinization purification had a certain effect on promoting L.f proliferation. Moreover, the PFSI gained by the third deproteinization purification, at a concentration of 10 g/L, significantly elevated L.f biomass and growth rate compared with the blank control, and the utilization of reducing sugars and the synthesis of protein and lactic acid were higher than the control (P < 0.05); improved LDH, SOD, and GSH-Px activity in L.f (P < 0.05), and the TCA method could be effectively applied to eliminate the proteins affecting FSI in probiotics, and PFSI may be a potentially beneficial prebiotic and intestinal reinforcer.

The Chemical Profiling and Anticancer Potential of Functional Polysaccharides from Flos Sophorae Immaturus.

Polysaccharides from Flos Sophorae Immaturus (FSI) are one of its pharmacological compounds that can perform effective activities. Aiming to extract the most effective polysaccharides against hepatocellular carcinoma (HCC), the polysaccharides were separated from FSI through ultrasonic microwave extraction, and the first comparison was carried out on the characterization of the structure and its cytotoxic properties on HCC SMMC 7721 cells of undeproteinized purified polysaccharides (PFSI-1) and papain-deproteinized polysaccharides (PFSI-2) from FSI. The findings indicated that PFSI-1 and PFSI-2 had characteristic absorption peaks of polysaccharides; PFSI-1 contained three monosaccharides and PFSI-2 contained ten; and SEM, AFM, and NMR were consistent with the verification of IR polysaccharide characteristics, suggesting probable additional latent activities. The pharmacotoxic effects of both PFSI-1 and PFSI-2 on SMMC 7721 cells (p < 0.05), attenuated the migration ability of SMMC 7721 cells (p < 0.05) and promoted apoptosis (p < 0.05), with an increase in G0/G1-phase cells and decrease in S-phase cells in the PFSI-1 as well as a decrease in G0/G1-phase cells, increase in S-phase cells, and decrease in apoptosis in the PFSI-2 (p < 0.05). The significant cytotoxic effect of PFSI-2 on SMMC 7721 cells (p < 0.05) and its protective effect on human hepatic L02 cells (HL-7702) at low concentrations (p > 0.05) could indicate its potential as a new drug for the treatment of HCC.

Biosynthesis and antibacterial activity of silver nanoparticles using Flos Sophorae Immaturus extract.

The current study proposes a green synthesis method for silver nanoparticles (Ag NPs) using various concentrations of Flos Sophorae Immaturus extract as reducing and capping agents. The UV-Visible (UV-Vis) spectroscopy, X-ray Diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR), X-ray Photoelectron Spectroscopy (XPS), Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM) were used to characterize resulting brown nanopowder. The as-prepared Ag NPs had a high negative zeta potential value of ∼ -38 mv, indicating the existence of electrostatic stabilization. The average sizes of ∼27.8 nm, 28.5 nm, 34.3 nm and 36.5 nm were measured by TEM. Moreover, FTIR and XPS analyses validated the production and chemical composition of Ag NPs from silver nitrate. The antibacterial activity of Ag NPs was examined against E. coli, P. aeruginosa, and S. aureus using agar well diffusion and the minimum inhibitory concentration (MIC) method. The antibacterial activity of the as-prepared Ag NPs from 4 mL extract was excellent against E. coli, P. aeruginosa, and S. aureus and the MIC values were 31.250, 15.625, and 31.250 mg/L, respectively. Based on these results, this study proposes a practical approach for the synthesis of Ag NPs in the industry and medical fields.

Styphnolobium japonicum (L.) Schott Flower Extract Alleviates Oxidative Stress and Inflammatory Factors in the Adjuvant-Induced Arthritis Rat Model.

INTRODUCTION: This research was to evaluate the beneficial effects of Styphnolobium japonicum (L.) Schott flower extract (SJF) on the adjuvant-induced arthritis rat model. METHODS: Arthritis was evoked by injection of complete Freund's adjuvant (CFA) in the hind paw. SJF (150 or 300 mg/kg/day) or Celecoxib (5 mg/kg/day) were administered intragastrically from the 0th day to the 28th day. The arthritis symptoms (paw edema, arthritic scores, mechanical hyperalgesia, and thermal hyperalgesia), inflammation biomarkers (RT and CRP), related enzymes (MMP1 and MMP13), oxidative stress markers (CAT, SOD, GPx, and MDA), and inflammatory cytokines (IL-6, IFN-γ, TNF-α, and IL-1ß) of SJF-treated CFA rats were evaluated. RESULTS: CFA rats exhibited severe arthritis symptoms, increased oxidative stress, and inflammatory cytokines. Interestingly, SJF treatment relieving arthritis symptoms and restored body weight gain compared with those in the CFA group. SJF treatment decreased the levels of CRP, RF, MMP1, and MMP13 in the CFA group. Besides, SJF treatment increased the activities of antioxidant enzymes and decreased the MDA content and inflammatory cytokines compared with those in the CFA group. Moreover, SJF could increase the mRNA expression of GPx-1 and CAT and inhibit the mRNA expression of IL-6 and TNF-α in the ankle tissue of CFA rats.

The Potential of Flos sophorae immaturus as a Pigment-Stabilizer to Improve the Monascus Pigments Preservation, Flavor Profiles, and Sensory Characteristic of Hong Qu Huangjiu.

Hong Qu Huangjiu (HQW) is distinguished by its inclusion of Monascus pigments, meaning that photosensitivity strongly affects the sensory quality of the wine. In this study, the effects of Flos sophorae immaturus (FSI) on the stability of Monascus pigments, the flavor profiles, and the sensory characteristics of HQW were investigated. After sterilization, the addition of FSI increased the preservation rate of Monascus pigments in HQW by up to 93.20%, which could be accounted for by the synergy of rutin and quercetin in FSI. The total content of the volatile flavor compounds in HQW increased significantly as the added amounts of FSI were increased, especially 3-methyl-1-butanol, 2-methyl-1-propanol, and short-chain fatty acid ethyl esters (SCFAEE). Sensory evaluation and partial least-squares regression revealed that the concentration of FSI significantly affected the aroma characteristics of HQW but had little effect on the mouthfeel. The addition of 0.9 mg/mL FSI yielded a satisfactory HQW with high scores in terms of mouthfeel and aroma. The strong correlation between fruit-aroma, full-body, and SCFAEE suggests that FSI might alter the aroma of HQW by enhancing the synthesis of SCFAEE. Summarily, treatment with FSI represents a new strategy for improving the stability of photosensitive pigments and thus adjusting the aroma of HQW or similar beverages.

Key aroma compounds and metabolic profiling of Debaryomyces hansenii L1-1-fermented Flos Sophorae.

The extract from Debaryomyces hansenii L1-1-fermented Flos Sophorae has a unique aroma and could be used as a natural spice. The influence of yeast growth and culture medium pH on organoleptic properties of fermented substrates, as well as on the content of volatile aromatic compounds, total sugars, polysaccharides, reducing sugars, total proteins, and amino acids, were analyzed. Metabolic pathways were annotated to compare and contrast key aromatic compounds and metabolic profiles of water and ethanol extracts of D. hansenii L1-1-fermented Flos Sophorae. We found that cells grew most rapidly, pH values changed significantly, and the largest consumption of sugars and amino acids occurred within 48 hr, producing bouquet-like, fruity, and sweet odors, as well as the highest content of volatile aromatic compounds in the extracts. The main aroma metabolites were 2-phenylethanol, linalool, and α-terpineol. The sensory quality of the ethanol extracts was superior to that of the water extracts. Five aromatic compounds, isoamyl alcohol, 2-methylbutan-1-ol, isobutyric acid, 2,3-hexanedione, and 1-hexanol, were positively correlated with the water extract group and negatively correlated with the ethanol extract group, whereas 13 aromatic compounds, styrene, acetophenone, 2-octen-1-ol, linalool, naphthalene, α-terpineol, dihydrocarveol, (-)-myrtenol, methyl anthranilate, eugenol, γ-nonanolactone, jasmone, and ß-ionone, showed the converse trend. Although 2-phenylethanol displayed the highest concentration in the extracts, it did not significantly contribute to the separation of ethanol and water extracts. In Flos Sophorae medium, D. hansenii mainly produces 2-phenylethanol from phenylalanine by the Ehrlich reaction, whereas it produces linalool and α-terpineol by the terpenoid backbone and monoterpenoid biosynthetic pathways; the variable contents of proline, arginine, and glutamate could respond to the arginine and proline metabolic pathways. PRACTICAL APPLICATIONS: Flos Sophorae, a collection of buds of Sophora japonica L., is a traditional Chinese medicinal and edible plant for its good aroma, taste, and nutritional value. Debaryomyces hansenii is a common, aroma-producing yeast. D. hansenii L1-1-fermented Flos Sophorae had a unique, bouquet-like aroma, slightly softer than the typical Flos Sophorae-like aroma. This study enriches our understanding of predominant aroma components and determines their contribution to the profiles of Flos Sophorae ferments obtained using D. hansenii L1-1. Researchers and manufacturers specializing in spices making can use these data to improve the aromatic profiles of natural spices produced by microorganisms, thereby obtaining unique aromas.

Combination of Flos Sophorae and chili pepper as a nitrite alternative improves the antioxidant, microbial communities and quality traits in Chinese sausages.

The main issue remains finding a nitrite alternative able to provide its multiple functions. Flos Sophorae exerts antioxidant and prebiotic actions, chili pepper has potent coloring capacity, thus this study investigated whether combination of Flos Sophorae and chili pepper could address the multiple activities of nitrite in Chinese sausages. Dry-fermented sausages were prepared: control and four treatments added with 150 mg/kg sodium nitrite (Nit), 0.2% Flos Sophorae (FS), 1% chili pepper (CP), and combination of 0.2% Flos Sophorae and 1% chili pepper (FS + CP). Results indicated that FS, CP and FS + CP had higher moisture, antioxidant activity and numbers of beneficial Staphylococcus and yeasts Candida, and lower numbers of Escherichia coli and harmful fungi, while FS had lower redness and harder texture than control. Their combination inhibited the declines of capsanthin and antioxidant capacity with ripening time, further improved microbiological communities compared with CP, and resulted in higher redness, similar color score and bacterial community, less lipid oxidation and softer texture compared with Nit. These results suggested that Flos Sophorae in combination with chili pepper could replace the nitrite's contribution to red curing color and microbiological communities, and effectively hinder lipid oxidation in Chinese sausages.

Evaluation of the synergetic effect of Yupingfeng san and Flos Sophorae Immaturus based on free radical scavenging capacity.

OBJECTIVE: This study aimed to determine the optimal extraction process and examine whether the combination of Flos Sophorae Immaturus (FSI) and Yupingfeng san (YPS) has a synergistic effect on free radical scavenging capacity. DESIGN AND METHODS: The time of immersion and extraction and the ratios (material/solvent) of the combination of YPS and FSI were optimized on the basis of polysaccharide and flavonoid yields via orthogonal design. The optimal result was used in the 1,1-diphenyl-1-picrylhydrazyl (DPPH) assay and animal experiments to test the antioxidant activity, which is reflected by superoxide dismutase, malondialdehyde, glutathione peroxidase, and total antioxidant capacity serum levels. The optimal extraction process was determined using various ingredients to obtain complex extracts with high active ingredient content and antioxidant activity. DPPH assay results showed that the optimized ingredients have antioxidant effects, and the combination had better antioxidation function than YPS in vitro. The combination also showed synergistic antioxidant activity compared with YPS in vivo. CONCLUSIONS: The combination of YPS and FSI had a synergistic antioxidant effect in vitro. The optimized extracts had antioxidant effects in vivo. These results indicated that YPS could be used with FSI to improve its antioxidant capacity in the body on the basis of free radical scavenging capacity.

Optimization of Ultrasound-Assisted Extraction Using Response Surface Methodology for Simultaneous Quantitation of Six Flavonoids in Flos Sophorae Immaturus and Antioxidant Activity.

Ultrasound-assisted extraction (UAE) was applied to extract rutin (RU), nicotiflorin (NI), narcissoside (NA), kaempferol (KA), isorhamnetin (IS), quercetin (QU), and total flavonoids of Flos Sophorae Immaturus (TFFSI) from Flos Sophorae Immaturus (FSI). Through single factor test and response surface methodology (RSM), the optimal extraction conditions were concluded as follows: ethanol concentration 70%, time 30 min, temperature 61 °C, and liquid/solid ratio 15.30 mL/g, respectively. The actual extraction rates of RU, NI, NA, KA, IS, QU, and TFFSI were 14.6101%, 2.9310%, 7.1987%, 0.1041%, 0.4920%, 2.7998%, and 26.4260%, respectively. The experimental results demonstrated that the extraction method with accuracy and efficiency could be used for the comprehensive evaluation quality control of extracts from FSI. The antioxidant activities of hydroalcoholic extraction from FSI on 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), superoxide anion (•O2-) free radicals, and ferric reducing/antioxidant power (FRAP) were assessed. The results showed that the antioxidation activities of extracts on DPPH, ABTS•+, and •O2- free radicals were reached 89.29%, 97.86%, and 56.61%, and 81.4% in FRAP at 1.0 mg/mL, respectively. The antioxidant capacity of FSI extract was positively correlated with the amount of total flavonoids.

Microwave-Assisted Extraction Combined with In-Capillary [Fe(ferrozine)3]2+-CE-DAD to Screen Active Components with the Ability to Chelate Ferrous Ions from Flos Sophorae Immaturus (Flos Sophorae).

An efficient microwave-assisted extraction (MAE) combined with in-capillary [Fe(ferrozine)3]2+-capillary electrophoresis-Diode Array Detector (in-capillary [Fe(ferrozine)3]2+-CE-DAD) was developed to screen active components with the ability to chelate ferrous ions and determine the total antioxidant activity. The MAE conditions, including methanol concentration, extraction power, extraction time, and the ratio of material to liquid, were optimized by an L9(34) orthogonal experiment. Background buffer, voltage, and cartridge temperature that affect the separation of six compounds were optimized. It was found that rutin and quercetin were the main components chelating ferrous ions in Flos Sophorae Immaturus (Flos Sophorae) by the in-capillary [Fe(ferrozine)3]2+-CE-DAD. The recoveries were ranged from 95.2% to 104%. It was concluded that the MAE combined with in-capillary [Fe(ferrozine)3]2+-CE-DAD method was a simple, reliable, and efficient tool for screening active components from the complex traditional Chinese medicine samples and evaluating their ability to chelate ferrous ions.

Efficient extraction of flavonoids from Flos Sophorae Immaturus by tailored and sustainable deep eutectic solvent as green extraction media.

In the face of the many shortcomings of conventional organic solvents in the age of green chemistry, deep eutectic solvents (DESs) appear under the spotlight of natural product extraction because of its outstanding advantages. In this study, the extraction of six compounds from Flos Sophorae Immaturus (FSI) with DES-5 (choline chloride/1,4-butanediol) as its topgallant solvent was determined by screening nine DESs. After single factor test and BBD experiment, the optimum conditions of deep eutectic solvent-based microwave-assisted extraction (DES-MAE) were: choline chloride/1,4-butanediol (molar ratio of 1:2) and water content (25%, v/v), time 20 min, microwave power 600 W, temperature 62 ℃, liquid/solid ratio 26 mL/g. The extraction yields of rutin, nicotiflorin, narcissin, quercetin, kaempferol and isorhamnetin were 116.78, 15.01, 23.85, 27.59, 3.09 and 3.33 mg/g, respectively. The kinetic experiment results showed that DES-MAE has significant advantages in the extraction of six compounds. The experimental results showed that DES-MAE could obtain higher yields of target components in a short time than other methods (DES-HRE, DES-UAE and Ethanol-MAE). In addition, the target components were separated from the DES extraction solution, and the recoveries of the target compounds were in the range of 75.5%-84.1%. Therefore, this paper provides a strategy for extraction and separation, the aim of which is to obtain flavonoids from FSI efficiently. Meanwhile, this study can also be used as an alternative to the traditional methods for obtaining bioactive components from plant sources in biochemistry, food industry and pharmaceutical fields.

[Effect of nitrogen, phosphorus and potassium fertilizers on Flos Sophorae Immaturus yield and quality in hilly area of Chongqing].

With Sophora japonica at the flowering stage as the object, the effect of nitrogen, phosphorus and potassium fertilizers on the yield composition factors, yield and quality of Flos Sophorae Immaturus (FSI) was studied. The results indicated that in early spring, nitrogen, phosphorus and potassium fertilizer on the amplification rate of S. japonica, FSI yield composition, yield and quality were different significantly, middle to high nitrogen (1.5-2.0 kg/plant) significantly increased the level of panicled clusters, raceme and flower bud number and yield. Phosphorus (1.5-2.0 kg/plant) could significantly increase the total buds of flower number and yield, potassium showed no significant increase in yield and yield components. Comprehensively considering yield and quality of FSI, nitrogen 1.5-2.0 kg/plant, phosphorus 1.5-2.0 kg/plant and potassium 0.6-0.9 kg/plant are appropriate.

Response Surface Optimized Infrared-Assisted Extraction and UHPLC Determination of Flavonoid Types from Flos Sophorae.

Single-factor experiment and Box-Behnken design were applied to optimize the infrared-assisted extraction (IRAE) of rutin, quercetin, kaempferol, and isorhamnetin from Flos sophorae. Four factors (extract solvent, solid-liquid ration, extraction time, infrared power) affecting the extraction yield of flavonoids were studied. Under optimized conditions the extraction yield was 33.199 ± 0.24 mg/g, which substantially improved, compared with heating reflux extraction (HRE) and ultrasonic-assisted extraction (UAE), while extraction time was only 9 min. The eluents were rich in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azobis (2-methyl-propionamidine) dihydrochloride radical scavenging potential (IC50 of DPPH: 53.44 ± 0.01 µg/mL, oxygen radical absorbance capacity (ORAC): 3785.83 ± 52 µmol/g) than the extracts obtained by HRE and UAE. In addition, an ultra-high performance liquid chromatography method was optimized for the identification and quantification of the tested flavonoids, and the method was validated based on its correlation coefficient (r), reproducibility (RSD, n = 5), and recovery values, which were 0.9994-0.9998, 0.74-1.83%, and 97.78-102.94%, respectively. These results confirmed that high extraction yield of flavonoids results in stronger antioxidant values and response surface methodology optimization of IRAE is a promising alternative to traditional extraction techniques for flavonoids from medicinal plants.

Bioactive flavonoids from Flos Sophorae.

Three new flavonoid glycosides-soyaflavonosides A (1), B (2), and C (3)-together with 23 known ones were obtained from the 70% EtOH extract of Flos Sophorae (Sophora japonica, Leguminosae). Their structures were elucidated by chemical and spectroscopic methods. Among the known isolates, 14, 18, 20, 22, and 26 were isolated from the Sophora genus for the first time; 12, 19, 24, and 25 were obtained from the species firstly. Moreover, NMR data for compounds 18 and 26 are reported for the first time here. Meanwhile, compounds 4, 8-13, 15, 16, 19, 21, and 22 presented obvious inhibitory effects on TG accumulation in HepG2 cells. Analysis of the structure-activity relationship indicated that all of the quercetin glycosides examined in this study possess significant activity that is not significantly influenced by the amount of glycosyl present, whereas increasing the amount of glycosyl reduced the activities of isorhamnetin glycosides and orobol. In addition, a high dose (30 µmol/l) of kaempferol was found to inhibit HepG2 cell growth, while a low dose (10 µmol/l) was observed to decrease TG accumulation.