Extract Isolation Purification and Identification of Polysaccharides from Exocarp of Unripe Fruits of Juglans mandshurica

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1 DOI /j. issn D101 DEAE Cellulose 52 Sephacryl S PJP - 1a PJP - 3a Da Da - PJP - 3a PJP - 1a PJP - 3a β PJP - 1a R A Extract Isolation Purification and Identification of Polysaccharides from Exocarp of Unripe Fruits of Juglans mandshurica YANG Hongxia 1 REN Xiaolei 1 HUO Jinhai 1 WANG Weiming Insititute of Chinese Materia Medica Heilongjiang Academy of Traditional Chinese Medicine Harbin Heilongjiang China 2. Center of Forect Resources Research and Development of Heilongjiang Harbin Heilongjiang China Abstract The purpose of this study was to extract isolate and purify polysaccharide from exocarp of unripe fruits of Juglans mandshurica analyze the monosaccharide composition and preliminarily analyze its structure of the obtained homogeneous molecular weight polysaccharide. A purified polysaccharide was obtained by hot water extractionethanol ZX RFQYJ zyyjy@ 163. com 10.. M M M M M M. 22. M M M M. 23. M M M M. 26. M M. 27. M M. 28. J

2 precipitation deproteinization and decolor. The purified polysaccharide then was isolated with DEAE Cellulose 52 column chromatography and Sephacryl S column chromatography. The purity identification and molecular weight determination were adopted by HPGPC. The monosaccharide composition and structure were analyzed by GC - MS and IR. The result showed that both the two obtained polysaccharides were homogeneous polysaccharide. The molecular weights of PJP - 1a and PJP - 3a were Da and Da respectively. PJP - 3a was mainly composed of rhamnose arabinose xylose mannose glucose and galactose at a molar ratio of IR suggested that both PJP - 1a and PJP - 3a had pyran ring structure and glycosidic bonds of PJP - 1a and PJP - 3a were both β type and PJP - 1a had acetamido structure. Key words Juglans mandshurica polysaccharide isolate purify analysis 120 Jusland emandehurica Maxim. 1 land emandehurica Maxim. 2 > 200 U mg - 1 Jus- 3 D101 DE- AE Cellulose 52 Whatman 4 5 Sephacryl S Pharmacia MG MG MG P D - B D - G D - M L - A L - S > 15 98% kg 14 1 h 14 1 h 4 95 % r /min 15 min Viskase - 100MG MG SIGMA - ALDRICH D g ml ph 6 3% 60 2 h 5 min D101 1 D101 1 h 2 BV h UV KD - 2KY DEAE Cellulose 52 RE - 52AA JM ml 3 ml DEAE Cellulose mol L - 1 NaCl LC - 10AS 2. 5 ml min ml / - Alltech ELSD 2000ES 620 nm PL aquagel - OH MIXED - H 8 μm GPC Spectrum Two 3500 Da 12 h Perkin Elmer Clarus SQ8T - 12 h PJP - 1 PJP - Perkin Elmer 3500 Da 2 PJP - 3 PJP - 4 PJP Sephacryl S mg 2. 5 ml Sephacryl S - 300

3 ml min - 1 min ml / nm 1. 2 ml min EI 70 ev m /z 43 ~ scan s - 1 PJP - 1a PJP - 3a mg 200 mg KBr ~ 4000 cm - 1 HPLC - GPC 2 mg 1 ml 2 mg 2 ml μm 2. 1 DEAE Cellulose 52 LC - 10AS HPLC aquagel - OH MIXED - H 8 μm GPC Alltech ELSD 2000ES 5 PJP mg ml μl mol L - 1 NaCl PJP mol L - 1 ml min L min - 1 NaCl PJP mol L - 1 NaCl PJP ml 2 mg ml μm 10 μl 2 mg ml - 1 T - PJP - 1 PJP - 3 logmw 2. 2 Sephacryl S PJP - 1 PJP - 3 Sephacryl S PJP - 1a PJP - 3a mg 4. 0 ml 2. 0 mol L - 1 TFA PJP - 1a PJP - 3a 3 h 4000 r /min 10 min PJP - 1a PJP - 3a TFA HPLC - GPC PJP - 1a PJP - 3a 15 min 2 PJP - 1a PJP ml 3a 60 mg 3 h PJP - 1a PJP - 3a 110 y = x R 2 = min ml 1. 0 ml h Da Da 5. 0 ml PJP - 3a μm GC - MS PJP - 3a L min L min D GC - MS 1 DB m mm μm min D min min5 min min min DEAE Cellulose mg 0. 5 mol L - 1 NaCl PJP PJP - 1 PJP - 3 PJP - 1a PJP - 3a HPLC - GPC min min min D min D PJP - 3a GC - MS 3 ~ DEAE Cellulose 52

4 PJP - 1a PJP - 3a HPLC - GPC 3 GC - MS PJP - 3a GC - MS PJP - 1a PJP - 3a 5 PJP 3-1a PJP - 3a 3400 cm - 1 Sephadex G100 Sephacryl S cm OH C - H PJP - 1a 1615 cm NH 2 COCH 3 C = O 27 PJP - 1a PJP - 3a cm - 1 PJP - 1a 2 mg ml μl 115 PJP - 3a 0. 2 mol L - 1 NaCl 1. 8 L min ml min - 1 C = O 1600 cm - 1 PJP - 1a cm - 1 β cm PJP GC - MS IR - 3a cm - 1 C - O - C 29 PJP - 3a cm - 1 β PJP - 1a PJP - 3a β PJP - 1a PJP - 3a

5 PJP - 1a PJP - 3a β PJP a J J J Smith J M. J J. J M. J Liu L Li W Sasaki T et al. Juglanone a novel α - tetralonyl derivative with potent antioxidant activity from Juglans mandshurica 21. J J. J Nat Med J J J J J. J Ruijun W Shi W Yijun X et al. Antitumor effects and immune 25. regulation activities of a purified polysaccharide extracted from Juglan regia J. Int J Biol Macromol LNT 2 - carboxylate interactions in metal - alginate complexes studied J with FTIR spectroscopy J. Carbohydrate Research J J J J Sup NF - κb J J J Papageorgiou S. K. Kouvelos E. P. Favvas E. P. et al. Metal. 29. J

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