To obtain the optimal process for the enzyme−assisted aqueous extraction of polysaccharides from Acanthopanax senticosus, and study the physicochemical properties of polysaccharides of different molecular weights, the extraction of Acanthopanax polysaccharides was optimized using the BBD response surface test. The polysaccharides with different molecular weights were obtained by ethanol−graded precipitation at 40%, 60%, and 80%, which were presented as ASPS40, ASPS60, and ASPS80. The polysaccharides were analyzed by HPGPC, ion chromatography, FT−IR, UV, SEM, TGA, XRD, Congo red, and I₂−KI tests. The antioxidant assay was used to evaluate their antioxidant properties in vitro. The findings demonstrated that the recovery rate of Acanthopanax polysaccharide was 10.53 ± 0.682%, which is about 2.5 times greater compared to the conventional method of hot water extraction. Based on FT−IR, TGA, polysaccharides with different molecular weights did not differ in their structure or thermal stability. The XRD suggests that the internal structure of ASPSs is amorphous. Congo red and I₂−KI showed that all three polysaccharides had triple helix structures with longer branched chains and more side chains. Furthermore, the antioxidant results showed the antioxidant activity of polysaccharides is not only related to the molecular weight size but also can be related to its composition and structure. These studies developed a green, and scalable method to produce polysaccharides from Acanthopanax senticosus and evaluated the properties of Acanthopanax polysaccharides of different molecular weights.

In order to improve the aqueous solubility and enhance the bioavailability of Hyperoside (Hyp), three inclusion complexes (ICs) of Hyp with 2-hydroxypropyl-β-cyclodextrin (2H-β-CD), β-cyclodextrin (β-CD), and methyl-β-cyclodextrin (M-β-CD) were prepared using the ultrasonic method. The characterization of the inclusion complexes (ICs) was achieved using Fourier-transform infrared spectroscopy (FTIR), scanning electronic microscopy (SEM), X-ray powder diffraction (XRPD), thin-layer chromatography (TLC), and ¹H nuclear magnetic resonance (¹H NMR). The effects of the ICs on the solubility and antioxidant activity of Hyp were investigated. A Job’s plot revealed that the Hyp formed ICs with three kinds of cyclodextrin (CD), all at a 1:1 stoichiometric ratio. The FTIR, SEM, XRPD, TLC, and ¹H NMR results confirmed the formation of inclusion complexes. The water solubility of the IC of Hyp with 2-hydroxypropyl-β-cyclodextrin was enhanced 9-fold compared to the solubility of the original Hyp. The antioxidant activity tests showed that the inclusion complexes had higher antioxidant activities compared to free Hyp in vitro and the H₂O₂–RAW264.7 cell model. Therefore, encapsulation with CDs can not only improve Hyp’s water solubility but can also enhance its biological activity, which provides useful information for the potential application of complexation with Hyp in a clinical context.

A green extraction method was developed using deep eutectic solvent extraction for the polysaccharide from Acanthopanax senticosus (A. senticosus). Among the eight types of DES prepared, the DES with a ratio of 1:4 L-malic acid to L-proline was found to be a suitable extraction solvent based on the extraction efficiency. The extraction parameters were optimized by Plackett–Burman and response surface methodology (RSM). The best extraction conditions were found for L-malic acid. Under the conditions of an L-malic acid/L-proline ratio of 1:4, ultrasonic power of 240 W, material–liquid ratio of 31.068 g/mL, water content of 32.364%, extraction time of 129.119 min, and extraction temperature of 60 °C, the extraction rate of A. senticosus polysaccharides was 35.452 ± 0.388 mg-g⁻¹. This rate was higher than that of polysaccharides obtained by hot water extraction (13.652 ± 0.09 mg-g⁻¹). The experimental results were best fitted by the quasi-secondary kinetic model when compared to two other kinetic models. Electron microscopic observations showed that DESs were more destructive to plant cells. The polysaccharide extracted from DESs had more monosaccharide components, a lower molecular weight, a higher antioxidant capacity, and superior anti-glycation activity compared to polysaccharides extracted from water (ASPS-PW). This study demonstrates the effectiveness of DESs in obtaining polysaccharides from A. senticosus.

Acanthopanax senticosus polysaccharide–nano-selenium (ASPS-SENPS) and A. selenopanax selenized polysaccharides (Se-ASPS) were synthesized, and their characterization and biological properties were compared. The acid extraction method was used to extract the polysaccharides of A. selenopanax, followed by decolorization using the hydrogen peroxide method and deproteinization based on the Sevage method, and the purification of A. senticosus polysaccharides (ASPS) was carried out using the cellulose DEAE-52 ion column layer analysis method. An A. senticosus polysaccharide–nano-selenium complex was synthesized by a chemical reduction method using ASPS as dispersants. The selenization of polysaccharides from A. selenopanax was carried out using the HNO₃-Na₂SeO₃ method. The chemical compositions, scanning electron microscopy images, infrared spectra, and antioxidant properties of ASPS-SENPS and Se-ASPS were studied, and they were also subjected to thermogravimetric analysis. The results indicated that the optimal conditions for the synthesis of ASPS-SENPS include the following: when ASPS accounts for 10%, the ratio of ascorbic acid and sodium selenium should be 4:1, the response time should be 4 h, and the reaction temperature should be 50 °C. The most favorable conditions for the synthesis of Se-ASPS were as follows: m (Na₂SeO₃):m (ASPS) = 4:5, response temperature = 50 °C, and response time = 11.0 h. In the in vitro antioxidant assay, when the mass concentration of Se-ASPS and ASPS-SENPS was 5 mg/mL, the removal rates for DPPH free radicals were 88.44 ± 2.83% and 98.89 ± 3.57%, respectively, and the removal rates for ABTS free radicals were 90.11 ± 3.43% and 98.99 ± 1.73%, respectively, stronger than those for ASPS. The current study compares the physiological and bioactivity effects of ASPS-SENPS and Se-ASPS, providing a basis for future studies on polysaccharides.

In order to optimize the process of solid-state fermentation and understand the changes of active ingredients during fermentation of A.senticosus, we used response surface methodology to optimize the solid-state fermentation process of A.senticosus and analyzed the effect of fermentation on the active components of A.senticosus by nontargeted metabolomics techniques. The effects of fermentation conditions on polyphenol content were studied in terms of fermentation time, fermentation temperature, water content, and the inoculum level through PlackettBurman design and response surface experiments. The optimized fermentation conditions were a fermentation time of 60h, a fermentation temperature of 37°C, a water content of 55, and an inoculum level of 12.5, and the polyphenol content reached 10.25±0.05mg·g⁻¹, which was 40 higher than that before fermentation. Furthermore, the metabolites and metabolic pathways of A. senticosus were analyzed before and after fermentation using nontargeted metabolomics techniques, and differences in metabolites were investigated based on the analysis of the principal components. The results showed that A.senticosus contained 57 phenolics, and a total of 12 polyphenolic metabolites were significantly different before and after fermentation (including phenols and isoflavonoids), among which the content of caffeic acid and its derivatives with antioxidant capacity was significantly increased, which may improve the antioxidant capacity of fermented A.senticosus. Therefore, solid-state fermentation opens new avenues for the clinical development and application of A. senticosus.

To enhance the nutritional value of Acanthopanax senticosus leaves (AL), a fermentation process was conducted using a probiotic Bacillus mixture, and the changes in chemical constituents and biological activities before and after fermentation were compared. A response surface methodology was employed to optimize the liquid fermentation conditions of AL based on their influence on polyphenol content. Non-targeted metabolomics analysis was performed using LC-MS/MS to reveal the differing profiles of compounds before and after fermentation. The results indicated that Bacillus subtilis LK and Bacillus amyloliquefaciens M2 significantly influenced polyphenol content during fermentation. The optimal fermentation conditions were determined to be a fermentation time of 54 h, a temperature of 39.6 °C, and an inoculum size of 2.5% (v/v). In comparison to unfermented AL, the total polyphenol and flavonoid contents, as well as the free radical scavenging capacities measured by DPPH and ABTS assays, and the activities of β-glucosidase and endo-glucanase, were significantly increased. The non-targeted metabolomics analysis identified 1348 metabolites, of which 829 were classified as differential metabolites. A correlation analysis between the differential metabolites of polyphenols, flavonoids, and antioxidant activity revealed that 13 differential metabolites were positively correlated with antioxidant activity. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis of the differential metabolites identified 82 pathways, with two of the top 25 metabolic pathways related to flavonoids. This study explores the potential for enhancing the active ingredients and biological effects of AL through probiotic fermentation using Bacillus strains.