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Ethanol volume fraction of 80% was widely applied for polysaccharides precipitation from Codonopsis pilosula water-leach liquor, while few researches focused on the dissolved components in 80% alcohol solution. Selenium nanoparticles presented strong biological activities with low toxicity, but the poor stability limited the practical applications. This study isolated a C. pilosula polysaccharide from the supernate after 80% ethanol precipitation, which was used as surface decorator for nano-composites preparation, and then the structural performance, storage stability and inhibitory mechanism on HepG2 cells were investigated. Results showed that the C. pilosula polysaccharide was mainly composed of fructose and glucose (molar ratio of 2.68:1.00) with average molecular weight of 3.21 × 103 Da. Additionally, the nano-composites were successfully prepared under Na 2 SeO 3 to polysaccharide (2.0 mg/mL) ratio (w:w) of 1:20, and exhibited homogeneous distribution with an average particle size of ~ 75 nm and superior stability at 4 °C for at least 35 days. Besides, the nano-composites effectively inhibited HepG2 cells proliferation dose-dependently, and induced the mitochondrial dysfunction via improving Bax expressions and descending Bcl-2 expressions, resulting in the release of mitochondrial cytochrome c to cytoplasm and the activation of caspase-9 / − 3, thereby promoting cells apoptosis. These results would effectively improve utilization rate of C. pilosula and provide theoretical foundation for clinical applications of the composite in hepatoma-bearing patients. • The novel fructose-enriched polysaccharide was prepared from Codonopsis pilosula. • The composites were prepared by selenium nanoparticles and the polysaccharide. • The prepared composites exhibited good dispersity and high stability. • The composites could effectively inhibit the proliferation of HepG2 cells. • The HepG2 cells were induced apoptosis mediated by caspase-9 and caspase-3. [ABSTRACT FROM AUTHOR]

The Codonopsis pilosula polysaccharide have been widely extracted through water dissolution and ethanol precipitation, and presented immunoregulatory and antioxidant activities. However, the polysaccharide from the supernatant was always discarded and rarely reported. In the present study, the extraction process, preliminary structural characteristics, immunoregulatory and antioxidant activities in vivo of C. pilosula polysaccharide prepared from the supernatant after 80% ethanol precipitation were evaluated and analyzed. Results showed that the optimized conditions for fructose-rich polysaccharide (Fru/Glc ratio of 2.73:1.00 with average molecular weight of 3.24 × 103 Da) extraction were determined as follows: liquid to material ratio of 40 mL/g, extraction time of 60 min, ultrasonic power of 370 W, and extraction temperature of 60 °C, and the yields reached to 6.52 ± 0.36%. Animal experiments demonstrated that the polysaccharide could effectively protect the immune organs, enhance the immune cells activities, and improve the leukocytes amounts and the expression levels of immune-related cytokines dose-dependently in CTX-treated mice, which was mainly attributed to the enhancement of antioxidant enzymatic activities and suppression of oxidative damage. These results suggested that the polysaccharide had the potential to be a natural antioxidant supplement due to the feasibility of industrial production and practical application. • The Codonopsis pilosula polysaccharide was isolated from 80% ethanol supernatant. • The maximum polysaccharide yield was 6.52 ± 0.36% optimized by Box-Behnken design. • The polysaccharide was a fructose-rich polysaccharide with molecular weight of 3.24 × 103 Da. • The polysaccharide exhibited strong immunoregulatory and antioxidant effects in vivo. [ABSTRACT FROM AUTHOR]

Cynanchum paniculatum has been applied for various diseases treatments, however, the polysaccharide components especially extracted using ethanol were rarely researched. In this study, ethanol solution with volume fraction of 75% was used for C. paniculatum polysaccharide preparation, and the extraction process optimization, structural characteristics, and bioactivities analysis were investigated. Results showed that extraction yield reached to 4.79 ± 0.24% under extraction temperature of 60 °C, extraction time of 4.5 h, liquid/solid ratio of 30 mL/g. Chemical constitutions and monosaccharide compositions, molecular weight determination results showed that the polysaccharide was mainly composed of glucose and xylose with a molar ratio of 1.00:0.63 (carbohydrate content of 94.91%, molecular weight of 5.1 kDa). Periodate oxidation, smith degradation, methylation, fourier transform infrared spectra and nuclear magnetic resonance analysis demonstrated that the glycosidic bonds configurations were mainly comprised of (1→2)-α- D -Glc p (39.86%), (1→2,4)-α- D -Glc p (10.32%) and (1→4)-β- D -Xyl p (29.58%). The bioactivities determination experiments results suggested that the polysaccharide exhibited strong antioxidant activities and antitumor effects both in vitro and in vivo. These data indicated that the ethanol-extracted polysaccharide from C. paniculatum holds great application prospect as a novel bioactive adjuvant in the future due to the feasibility of industrial production. • The polysaccharide was extracted from Cynanchum paniculatum using 75% ethanol. • The maximum extraction yield was 4.79 ± 0.24% optimized by Box-Behnken design. • The polysaccharide was composed of glucose and xylose (molar ratio of 1.00:0.63). • The polysaccharide was comprised of (1→2)-Glc p (39.86%) and (1→4)-Xyl p (29.58%). • The polysaccharide exhibited strong antioxidant and antitumor effects. [ABSTRACT FROM AUTHOR]

• The novel polysaccharides were extracted from Salvia miltiorrhiza by 60 % ethanol. • The proper extraction conditions for this crude polysaccharides were optimized. • The polysaccharides (6.50 × 103 Da) was composed of Gal/Glc/Fru (1.00:0.45:0.28). • The polysaccharides could notably improve immune levels of immunocompromised mice. Salvia miltiorrhiza polysaccharides were generally extracted using distilled water as solvent and showed various bioactivities in vivo , however, the preparation, structure, and immunomodulatory effects of polysaccharides with 60 % ethyl alcohol extraction were rarely reported. In the present study, S. miltiorrhiza polysaccharides was prepared using 60 % ethanol and the extraction process was optimized, then the structural characteristics and immunomodulation activity were investigated. Results showed that optimal extraction conditions for crude polysaccharides were determined as follows: liquid/solid ratio of 22 mL/g, extraction temperature of 60 °C, ultrasonic power of 205 W, ultrasonic time of 125 min, and the yields reached to 5.99 ± 0.09 %. Structural determination results demonstrated the purified polysaccharides showed a average molecular weight of 6.50 × 103 Da with Galactose/Glucose/Fructose ratio of 1.00:0.45:0.28. Animal experiments indicated that the polysaccharides could effectively enhance immune cells activities, and improve lymphocytes subsets proportions and immune-related cytokines expression levels dose-dependently in cyclophosphamide-treated mice. These results would provide a theoretical basis for industrial production and practical application of the novel polysaccharides as an immunoregulation adjuvant. [ABSTRACT FROM AUTHOR]