Background: Polygonatum cyrtonema, one of origins of Polygonata Rhizoma (HuangJing in Chinese), is traditionally steamed repeatedly before being used as herbal medicine in China. However, there had no standardization of steaming of HuangJing. Therefore, comprehensive study for effects of steam on polysaccharide from Polygonatum cyrtonema based on saccharide mapping, a powerful method developed for polysaccharides analysis, and pharmacological activity are still necessary in order to explore the effect of steam on the physiochemical and biological activities of its polysaccharides and determine the standard for Polygonatum cyrtonema.Methods: To explore the effect of steam on the physiochemical and biological activities of P. cyrtonema polysaccharides (PCP), six polysaccharides named PCP0, PCP1, PCP2, PCP3, PCP4 and PCP5 were extracted from the herb consecutively steamed for 0 to 5 times, respectively. Their molecular weight distribution, monosaccharide composition and PACE fingerprints were investigated through HPSEC-MALLS-RID, HPAEC-PAD and saccharide mapping based on polysaccharide analysis by using carbohydrate gel electrophoresis (PACE) and HPTLC, respectively. In addition, their antioxidant ability and immunostimulatory activities on RAW 264.7 cells in term of NO production and phagocytosis were compared.Results: Results suggested that molecular weights could be changed during steam, which increased by first steaming and then decreased with further steaming though all polysaccharides molecular weight were 105-107 Da. They all showed irregularly spherical conformation in aqueous solution based on AFM imaging. Monosaccharide composition and PACE fingerprints was significantly different after steaming, i.e., galactose increased while glucose and mannose decreased, and β-1,4-Galp appeared while β-1,4-manp increased after steaming. Steamed PCP significantly increased scavenging activity against ABTS radicals, while PCP0 had better immunostimulatory effect on RAW 264.7 in terms of NO production and phagocytosis.Conclusions: In summary, steam significantly affected the chemical composition and bioactivities of polysaccharides from P. cyrtonema. Considering the conflict results of steaming on antioxidant and immunopotentiation activities of PCP, 2 times of continuously steam is the optimal choice under the modern process condition.
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