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1 6 8 Vol. 6 No Journal of Food Safety and Quality Aug., 2015 * 茹翌, 徐光勋, 赵欢, 陈红漫 (, ) 摘要 : 目的, 方法 (Momordica charantia polysaccharide, MCP), DEAE-52 4 (MCP Ⅰ, MCP Ⅰ, MCP Ⅰ, MCP Ⅰ) MCPⅠ CMMCPⅠ MCPⅠ-Ca( Ⅰ), Fenton OH 结果 MCPⅠ MCPⅠ-Ca( Ⅰ ) CMMCPⅠ, 0.7 mg/ml, OH 58% 82% 93%, SC mg/ml,, 结论关键词 : ; ; ; Modification of polysaccharide from Momordica charantia L. and its antioxidant activities RU Yi, XU Guang-Xun, ZHAO Huan, CHEN Hong-Man * (College of Bio-Science Techonlogy, Shenyang Agricultural University, Shenyang , China) ABSTRACT: Objective To investigate the antioxidant activities of modified polysaccharides, which was embellished by carboxy methylated and metal ions. Methods Water soluble crude polysaccharide was extracted from Momordica charantia L. with hot water and precipitated by ethanol, then purified by DEAE-cellulose ion-exchange chromatography with NaCl gradient elution to obtain 4 kinds of polysaccharide components (MCP Ⅰ, MCP Ⅰ, MCPⅠ, MCP Ⅰ). The antioxidant activities of MCPⅠ were tested by determining scavenging rates of the fraction on OH in vitro. The calcium-polysaccharide complex was synthesized when chelated with Ca 2+, coded as MCPⅠ- Ca( Ⅰ ). The carboxy methylated polysaccharide was named as CMMCPⅠ. Results The MCP Ⅰ, MCPⅠ-Ca( Ⅰ) and CMMCP Ⅰ showed obvious antioxidant activities to OH. Their scavenging rates were 58%, 82% and 93% at a mass concentration of 0.7 mg/ml, respectively. The SC 50 values in the study were less than 0.6 mg/ml. Furthermore, CMCPⅠ exhibited the strongest antioxidant activities among them. Conclusion The current study suggests that carboxy methylated and metal ions can improve polysaccharide bearing excellent antioxidant activities. KEY WORDS: Momordica polysaccharide; metal complexation; carboxy methylation; antioxidant activity 基金项目 : ( ) ( ) Fund: Supported by the Natural Science Foundation of China ( ) and the College Student Innovation Foundation of Shenyang Agricultural University ( ) * 通讯作者 :,, hongmanc@hotmail.com *Corresponding author: CHEN Hong-Man, Associate Professor, Shenyang Agricultural University, No.120, Dongling Road, Shenhe District, Shenyang , China. hongmanc@hotmail.com

2 引言 (Momordica charantia L.),,,,,,,, [1-4], [5], [6,7],,,, [8,9],,, [12-14], 2, 2 材料与方法 2.3 实验方法苦瓜多糖的分离流程 50 g (,, 40 ) ( 1:40, 3 mg/g, 50 5 h) (3500 r/min, 10 min) ( 1/3, 3 95% ) (3500 r/min, 10 min) Sevage 苦瓜多糖的纯化 0.1 g, 50 ml (50, 30 min), 10 min, (3500 r/min, 10 min),, DEAE-52, NaCl mol/l 多糖羧甲基化修饰及取代度的测定 0.1 g MCPⅠ, 10 ml, 30 min, 20% NaOH 2 ml, 50 min 25 mg 30 min, 60 3 h,, ph 5.0; 3 d, MCP Ⅰ, CMCPⅠ [5] CMCPⅠ10 mg, 0.2 mol/l HCl (70% )4 ml, 3 h, 80%, 0.01 mol/l NaOH, 2.1 仪器与设备 0.01 mol/l HCl, TGL-10C ( [6,7] ); HH-6 ( ); CNaOHVNaOH CHClVHCl 0.162A A W A N-1001 ( ); : C NaOH =0.01mol/L; C HCl =0.01mol/L ; W SpectrAA-220 ( JENA (mg); A NaOH ), Nexus670FT-IR Nicolet 苦瓜多糖钙络合物的制备及取代度测定 ; ALPHA 2-4LD ( CHRIST ); 40 mg MCPⅠ, 20 ml Unic7200 ( ) ),, 5:1 2.2 材料与试剂, 60, 1 h,, 1/3, 3 ( );,, DEAE-Cellulose(DE-52, Sigma ), (DPPH, Bio Basic ); 48 h, 48 h( 95% ( ), 3~5 ),,,, MCPⅠ-Ca(II)

3 8, : 2919 MCPⅠ-Ca(II) 20 mg,, 500~550 2 h,,, [8] 羟基清除率的测定 Fenton OH, 4.5 mmol/l FeSO mmol/l mol/l,, MCPⅠ H 2 O 2 1 ml, H 2 O 2, 37 1 h,, 510 nm P%=(A 0 -(A x -A x0 ))/A 0 *100% : A 0 : ; A x : ; A x0 : [9] DPPH 清除率的测定 2 ml 75% DPPH 2 ml, 1 h, 510 nm (%)=(1-(Ai-Aj)/ A 0 )*100% : Ai DPPH ; Aj ; A 0 DPPH [10] 红外光谱分析 MCPⅠ-Ca(II) CMMCPⅠ 1 mg, KBr, : 4000~500 cm -1 3 结果与分析 3.1 多糖的分离纯化 Sevage, 100 mg, DEAE-52, 1,,, MCP Ⅰ; 0.1 mol/l NaCl, MCP Ⅰ; 0.3 mol/l NaCl, MCPⅠ; 0.5 mol/l NaCl, MCPⅠ MCPⅠ ( OH), 65%, 3 1 MCPs DEAE- Fig. 1 Elution curve of MCPs on DEAE-cellulose column 2, 500~4000 cm ~3100 cm -1 -OH, COOH C=O ; COO C=O ; COOH C-O ; COO C=O ; OH ; C-O-C ; C-O ; β-d- β ; β-d, [11,12] C H ; 苦瓜多糖羧甲基化修饰研究 MCPⅠ, CMCPⅠ MCPⅠ, 3, CH 2 -CH, cm cm -1, COO-,, [13] 950~1200 cm -1 C-O-C C-O-H [14] 苦瓜多糖金属络合物制备, MCPⅠ-Ca( Ⅰ), 4 4

4 Fig. 2 2 MCPⅡ IR spectrum of the polysaccharide MCPII Fig. 3 3 CMCPⅡ IR spectrum of the polysaccharide CMCPII cm -1 (O-H),, ( cm cm -1 ),, [15],, 3.4 苦瓜多糖及其抗氧化活性

5 8, : 2921, Fenton DPPH 5, Fenton, 0.7 mg/ml, MCPⅠ-Ca( Ⅰ) MCPⅠ 28%, CMMCPⅠ 40% SC 50 : MCPⅠ 0.592, MCPⅠ-Ca( Ⅰ) 0.548, CMMCPⅠ 0.358, : MCP Ⅰ > MCPⅠ-Ca( Ⅰ)>CMMCPⅠ DPPH 6, 0.7 mg/ml, MCPⅠ-Ca( Ⅰ) DPPH MCPⅠ 14%, CMMCPⅠ DPPH 24% SC 50 : MCPⅠ 0.507, MCPⅠ-Ca( Ⅰ) 0.450, CMMCPⅠ 0.413, MCP Ⅰ>MCPⅠ-Ca( Ⅰ )> CMMCPⅠ 4 Fig. 4 The IR spectra of MCPⅡ-Ca(Ⅱ) Fig. 5 5 MCPⅡ CMMCPⅡ, MCPⅡ-Ca(Ⅱ) Hydroxyl scavenging ( OH) effect of MCPⅡ, CMMCPⅡand MCPⅡ-Ca(Ⅱ)

6 MCPⅡ CMMCPⅡ, MCPⅡ-Ca(Ⅱ) DPPH Fig. 6 DPPH radical scavenging effect of MCPⅡ, MCPⅡ-Ca(Ⅱ) and CMMCPⅡ 4 讨论,,,,,,,,,, [8],,,,,, 参考文献 [1] Ahmed I, Lakhani MS, Gillett M, et al. Hypotriglyceridemic and hypocholesterolemic effects of anti-diabetic Momordica charantia (karela) fruit extract in streptozotocin-induced diabetic rats [J]. Diabetes Res Clin Pr, 2001, 51(3): [2] Wu SJ, Li TN. Antioxidant and free radical scavenging activities of wild bitter melon (Momordica charantia Linn. var. Abbreviata Ser.) in Taiwan [J]. LWT Food Sci Technol, 2008, 41: [3] Sandhya LS, Shewade Y, Bhonde R. Role of bitter gourd fruit juice in stz-induced diabetic state in vivo and in vitro [J]. J Ethnopharmacol, 2000, 73: [4] Seham AE, Souad EE, Osama AE. Some toxicological studies of Momordica charantia L. on albino rats in normal and alloxan diabetic rats [J]. J Ethnopharmacol 2006, 108: [5],,,. [J]. ( ), 2012, 33(6): Qu SL, Wang YC, Luo L, et al. Carboxymethyl modification effect on the anti oxidant activity of jujube polysaccharide [J]. J Henan Univ Technol (Nat Sci Ed), 2012, 33(6): [6] Eyler RW, Klug ED, Diephuis F. Determination of degree of substitution of sodiu m carboxymethylcellulose [J]. Anal Chem, 1947, 19 (l): [7]. [D]. :, Liang J. Study on the chemical modification of tea polysaccharide and its anti coagulation effect in vitro [D]. Hefei: Anhui Agricultural University, [8] Yang LQ, Zhao T, Wei H, et al. Carboxymethylation of

7 8, : 2923 polysaccharides from Auricularia auricula and their antioxidant activities in vitro [J]. Inter J Biol Macromol, 2011, 49: [9] Niu YG, Wang HY, Xie ZH, et al. Structural analysis and bioactivity of a polysaccharide from the roots of Astragalus membranaceus (Fisch) Bge. var. mongolicus (Bge.) Hsiao [J]. Food Chem, 2011, 128: [10] Parikh A, Madamwar D. Partial characterization of extracellular polysaccharide from cyanobacteria [J]. Biores Technol, 2006, 97: [11] Coimbra MA, Goncalves F, Barros AS, et al. Fourier transform infrared spectroscopy and chemometric analysis of white wine polysaccharide extracts [J]. J Agric Food Chem, 2002, 50: [12] Manrique GD, Lajolo FM. FT-IR spectroscopy as a tool for measuring degree of methyl esterification in pectins isolated from ripening papaya fruit [J]. Posth Biol Technol, 2002, 25: [13] acuráková M, Capek P, Sasinková V, et al. FT-IR study of plant cell wall model compounds: pectic polysaccharides and hemicelluloses [J]. Carbohyd Poly, 2000, 43: [14]. [J]., 2011, 38(2), 78, 91. Zhong Z. Summary of momordica charantia polysaccharides [J]. Guangdong Chem Ind, 2011, 38(2), 78, 91. [15],,,. [J]., 2000, 20(2): Yang LM, Weng SP, Yang LQ, et al. Several mono and disaccharides and some sugar solid metal complexation [J]. Spectr Spectr Anal, 2000, 20(2): 作者简介 ( 责任编辑 : 杨翠娜 ) 茹翌, 本科生, 主要研究方向为生物活性大分子 @qq.com 陈红漫, 博士, 副教授, 研究生导师, 主要研究方向为生物活性分子的结构与功能, 分子酶学与酶工程 hongmanc@htomail.com

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核农学报２０１６３０５０９２００９２５ＪｏｕｒｎａｌｏｆＮｕｃｌｅａｒＡｇｒｉｃｕｌｔｕｒａｌＳｃｉｅｎｃｅｓ９２０文章编号１０００８５５１２０１６０５０９２００６羊栖菜活性组份多糖的体外抗氧化能力比较丁浩淼１２李伟强１汪财生 1 顾筱筱１谢琰１钱国英１王忠华１１浙江万里学院生物与环境学院浙江宁波３１５１００２浙江海洋高效健康养殖协同创新中心