Arabinose

Glucose (Glu), Fructose (Fru), Mannose (Man), Galactose (Gal), Xylose (Xyl), Arabinose (Ara), Rhamnose (Rha), Fucose (Fuc), Galactose Uronic acid (GalUA), glucuronic acid (GlcUA), purity ≥ 98%, β-dextran (purity ≥ 80%), fructo oligosaccharide (purity ≥ 95%), xylan (purity ≥ 98%) and polygalacturonic acid (purity ≥ 90%) were purchased from Yuanye Biotechnology Co., Ltd. D-trehalose (purity ≥ 99%) was purchased from Shanghai McLean Biochemical Technology Co., Ltd. All of other chemicals and reagents were analytical grade.

The medicinal materials in this study were the cultivated C. pilosula, which were harvested during the harvest period. They were identified as Codonopsis pilosula (Franch.) Nannf by Professor Gu Wei of Nanjing University of Chinese Medicine. The voucher samples and digital image information were stored in the Herbarium of Nanjing University of Chinese Medicine.
Dried roots of C. pilosula were purchased from Pingshun County, Shanxi province, China. The herbs were dried at 50 °C, cut into 1 cm strips, and crushed into powder that passed through a sieve with 80 mesh. Mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, xylose, arabinose, fucose (all ≥ 98.0% purity), papain, and 1-phenyl-3-methyl-5-pyrazolone (PMP) were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. Trifluoroacetic acid (TFA) and other chemicals were purchased from Shanghai Macklin Biochemical Co., Ltd. Acetonitrile (HPLC grade) was obtained from Merck Darmstadt Ltd. (Germany). All chemicals were reagent grade or better.

Fresh pumpkin peel was obtained from Laiyang Mengyu Co., Ltd. (Yantai, China). CrCl₃·6H₂O (analytical grade) was obtained by Tianjin Kaitong chemical reagent Co., Ltd. (Shanghai, China). Trifluoroacetic acid (TFA) and insulin were provided by Sigma-Aldrich Co., Ltd. (St. Louis, MO, USA). α-Glucosidase, 4-nitrophenyl-α-D-glucopyranoside (PNPG), penicillin-streptomycin, and albumin bovine were purchased from Beijing Solarbio Technology Co., Ltd. (Beijing, China). Acarbose and monosaccharide standards (rhamnose, arabinose, galactose, glucose, mannose, xylose, fructose, galacturonic acid, and glucuronic acid) were obtained from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Sodium acetate, anhydrous (electrochemical grade), and SS254-500 sodium hydroxide solution were purchased from Thermo Fisher Scientific (Sunnyrale, CA, USA). High-glucose DMEM medium was obtained from Wuhan Servicebio Technology Co., Ltd. (Wuhan, China). Fetal bovine serum was purchased from Zhejiang Tianhang Biotechnology Co., Ltd. (Huzhou, China). Trivalent chromium standard solution was supplied by Beijing Century Aoke Biotechnology Co. Ltd. (Beijing, China). All other chemicals and reagents used were of analytical grade.

Mannose (B25303), rhamnose (YY91163), galacturonic acid (B21894), glucose (B21882), galactose (YY91011), and arabinose (B25845) were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. PMP (1-phenyl-3-methyl-5-pyrazolone) pre-column derivatization combined with high performance liquid chromatography (HPLC) were used to determine the monosaccharide compositions of CSP (Cao et al., 2019 (link)). The polysaccharide (1.0 mg/ml) was hydrolyzed using 4 M trifluoroacetic acid (TFA) at 110°C for 6 h. The residual TFA was obtained by evaporating the residue under reduced pressure in methanol (2 ml). The hydrolysate and the hybrid standard monosaccharide solution (100 μl, glucose, D-mannose, D-galactose, xylose, maltose, L-arabinose, all 1 mg/ml) were dissolved in 200 μl water; the supernatant was mixed with PMP and 0.3 M NaOH solutions. The reaction was carried out at 70°C for 45 min before being stopped using an HCl solution. The reaction product was removed thrice with chloroform, and the upper layer was centrifuged, and filtered using a 0.22 μM membrane, followed by HPLC analysis. We used the Agilent Eclipse XDB-C18 chromatographic column (30°C), equipped with a UV-detector. The mobile phase was PBS (pH 6.8) mixed with acetonitrile (87: 13) and eluted isocratically for 60 min.

Dried fruiting bodies of T. mongolicum Imai were obtained from Zhangjiakou, Hebei Province, China. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) was purchased from Solarbio Science and Technology Co., Ltd. (Beijing, China). The total antioxidant capacity assay kit for the ABTS method was purchased from Beyotime Co., Ltd. (Shanghai, China). Monosaccharide standards (mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, N-acetyl-D-glucosamine, N-acetyl-D- galactosamine, glucose, galactose, xylose, arabinose, and fucose) were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). All the reagents and chemicals used in this study were analytical or HPLC grade.

Hemerocallis citrina Borani was obtained from Qingyang, Gansu, China. Standards of glucose, mannose, fucose, rhamnose, xylose, galactose, arabinose, glucuronic acid, ribose, and galacturonic acid were purchased from Shanghai Yuanye Biotechnology (Shanghai, China). Absolute ethanol was purchased from Fuyu Fine Chemical (Tianjin, China). Trifluoroacetic acid, methanol, sodium hydroxide, hydrochloric acid, carbazole, 1-phenyl-3-methyl-5-pyrazolone, 3,5-dinitrosalicylic acid, sulfuric acid, Coomash bright blue, chloroform, potassium bromide, sodium azide, sodium acetate, anthranone, and other reagents were purchased from Solarbio Biotechnology (Beijing, China). ABTS and FRAP assay kits were purchased from Suzhou Keming Biotechnology (Suzhou, China). Distilled water was used in these experiments.

The Cell Bank of the Chinese Academy of Sciences (Beijing, China) provided the murine macrophage cell line RAW 264.7 for use in research. Trimethylchlorosilane, hexamethyldisilane, pyridine, and trifluoroacetic acid (TFA) were obtained from Aladdin Biological Technology Co., Ltd. (Shanghai, China). Rhamnose, arabinose, xylose, fucose, mannose, glucose, and galactose were purchased from YuanYe Bio-Technology Co., Ltd. (Shanghai, China). Lipopolysaccharides (LPSs), Brain Heart Infusion (BHI), Mw cut-off 8000–14,000 Da MWCO membranes (MD34), and 0.1% neutral red stain solution were bought from Solarbio Life Science and Technology Co. Ltd. (Beijing, China). Bovine calf serum was purchased from Sigma Chemical Co., Ltd. (Saint Louis, MO, USA). ELISA kits for the analysis of TNF-α and IL-6 were purchased from Neobioscience Technology Co., Ltd. (Shenzhen, China). Cell Counting Kit-8 (CCK-8) was purchased from Beyotime Biotechnology Co. Ltd. (Shanghai, China). A kit for measuring nitric oxide (NO) was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).