白术多糖表征与免疫活性研究.doc

SUPPLEMENTARY MATERIAL Characterization of oligosaccharides from Baizhu by HILIC-MS Zhe Lina?, Yan-Fang Liub?, Yang Qua, Li-Ying Shib, De-Qiang Doua,?The first two authors contributed equally to this article.? ?The first two authors contributed equally to this article. ? Corresponding author. De-Qiang Dou: deqiangdou@126.com Hai-xue Kuang: kuanghx@ a College of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600, China b Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China c Heilongjiang University of Chinese Medicine, Harbin 150000, China Abstract Polysaccharides from Baizhu was separated by prepatative hydrophilic interaction liquid chromatography (HILIC) on a XAmide column and its components was characterized as inulin-type polysaccharides with structures of α-D-glucopyranosyl-[-(1→2) -β-D-fructofuranosyl-]n-1-(1→2) -β-D-fructofuranoside (n=3-20) by a combinatory application of ESI-MS, NMR and IR, as well as the chemical analysis of monosaccharide composition. In addition, the contents of nystose and 1F-fructofranosylnystose in the crude and purified Baizhu polysaccharides were determined to be 5.81%, 4.92% and 0.70%, 0.84% (w/w) respectively. In addition, MTT assay indicated that the Baizhu polysaccharides could effectively promote spleen lymphocyte transformation for the enhancement of organism immunity. It is for the first time that inulin-type polysaccharides were discovered in Baizhu and its immunoenhancing activity was reported, which is an vigorous evidence to explain the efficicacy of Baizhu. 1. Experimental 1.1. Materials and reagents XAmide (280 mm × 50 mm I.D., 10 ?m; 250 mm × 4.6 mm I.D., 10 μm; 150 mm × 4.6 mm I.D., 5 μm ) were made from Acchrom Technology Co., Ltd., Beijing, China and the column of Agilent ZORBAX NH2 (250 mm × 4.6 mm I.D., 5 μm) was purchased from Agilent Technologies Co., Ltd. (USA). Acetonitrile (Tedia Company Inc., USA) was chromatographic grade and other reagents