7 3 Vol. 7 No. 3 2016 3 Journal of Food Safety and Quality Mar., 2016 陈梅春 1, 张海峰 2, 陈峥 1, 潘志针 1, 朱育菁 1, 刘波 (1., 350003; 2., 350003) 1* 摘要 : 目的,, DPPH 方法 (WTPS-1), ; ; - (GC-MS) ; DPPH 结果 WTPS-1 β-, 71.9%,, 2.57:3.67:0.57:1:2.16:9.65 WTPS-1 DPPH 91.6% 60.7% 9.20% 结论 WTPS-1, 关键词 : ; ; ; DPPH Isolation purification and composition analysis of Oolong tea polysaccharides CHEN Mei-Chun 1, ZHANG Hai-Feng 2, CHEN Zheng 1, PAN Zhi-Zhen 1, ZHUYu-Jing 1, LIU Bo 1* (1. Agricultural Bio-resources Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China; 2. Fujian Academy of Agricultural Sciences, Fuzhou 350003, China) ABSTRACT: Objective To isolate, purify and analyze the composition of Oolong tea polysaccharides, and to evaluate the DPPH radical scavenging activity of polysaccharides. Methods Tea polysaccharids fraction (WTPS-1) was isolated and purified from Oolong tea using ion exchange chromatography, the neutral sugar of WTPS-1 was measured by phenol-sulfuric acid method, the structure of WTPS-1 was analyzed by infrared spectroscopy (IR), the monosaccharide composition of WTPS-1 was measured by gas chromatography-mass spectrometer (GC-MS), and the antioxidant activity of WTPS-1 was evaluated by DPPH assay. Results The polysaccharide WTPS-1 contained β-glycoside bond type pyranose, and the content of neutral sugar was 71.9%. The monosaccharide compositions were Rha, Ara, Xyl, Man, Glu and Gal with a ratio of 2.57:3.67:0.57:1:2.16: 9.65. DPPH scavenging activity of crude polysaccharide, deproteinized polysaccharide and WTPS-1 were 91.6%, 60.7% and 9.20%, respectively. Conclusion The crude polysaccharide exhibited the relative stronger DPPH scavenging activity than deproteinized polysaccharide and WTPS-1, and the results would provide useful references for the development of Oolong tea. KEY WORDS: polysaccharide; structure composition; monosaccharides components; DPPH scavenging activity 基金项目 : (2015NZ0003) (YC2015-11) Fund: Supported by the Fujian Key Science and Technology Special Projects (2015NZ0003) and the Youth Fund of Fujian Academy of Agricultural Sciences (YC2015-11) * 通讯作者 :,, E-mail: fzliubo@163.com *Corresponding author: LIU Bo, Researcher, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China. E-mail: fzliubo@163.com
3, : 1299 1 引言,,, [1-4],,,,,,,,,, [5] OH O.- 2, ; Chen [6],, ; Chen [7], ; Chen [8], ; Yang [9],,,,,,,, Sevag,, 2 材料与方法 2.1 仪器与试剂 DK-8D ( ) GL10MD ( ) ABZ1-1001-P ( ) UV-2550 ( Shimadzu ) Nicolet Magana750 FT-IR ( Nicolet ) 7890A-5975C ( Agilent ) ( ) ; 1, 1- (DPPH) ; ( ) G-250 DEAE DE-52 : 2.2 茶多糖提取, 20 ml, 50 1 h,,,, 3 95%,, 2.3 茶多糖脱蛋白工艺, Sevag Sevag ( : =4:1, V:V), 20 min, 5 min,,,, 3, 3 95% 2.4 乌龙茶多糖的分离纯化 DEAE-52, φ2.6 cm 50 cm, 0.1 mol/l 0.2 mol/l 0.3 mol/l NaCl, 1 ml/min,, 3 min 1, -,,,, 2.5 中性糖含量的测定 -, 3 2.6 茶多糖清除 DPPH 自由基能力的测定 0.2 mg/ml, 0.2 mmol/l DPPH 2 ml,, 30 min, 50%
1300 7, 525 nm A i, 2.0 ml 60% 525 nm A j, 2.0 ml DPPH 2.0 ml 60% 525 nm A 0, : (%)= [1 (A i A j )/A 0 ] 100 3 200~400 nm, 2 200~400 nm 2.7 紫外可见光谱 1.0 mg/ml DEAE-52,, 200~400 nm 2.8 红外光谱 DEAE-52 KBr,, 4000~400 cm -1 2.9 茶多糖的气相色谱 - 质谱分析 : WTPS-1 20 mg 2 ml 2 mol/l (TFA),, 100 6 h, TFA : 20 mg 1 ml, 90 30 min, 1 ml, 90 30 min,,, 0.5 ml GC-MS : HP-5MS, 270, 17.8 kpa, 10:1, 3 ml/min, 1 μl, : 150 6 min, 2 /min 220, 2 min : EI; ; 6 min; : 25.00~550.00 amu; EMV ; MS 230, MS 150 3 结果与分析 3.1 茶多糖的 DEAE-52 纤维素柱层析 DEAE-52, 1, 0.1 0.2 0.3 mol/l NaCl, 3,,, (WTPS-1), 3.2 乌龙茶 WTPS-1 多糖的紫外和红外光谱分析 0.4 mg/ml WTPS-1, 1 DEAE-cellulose Fig. 1 Chromatographic isolation of tea polysaccharides by DEAE-cellulose column Fig. 2 2 WTPS-1 UV spectrum of Oolong tea polysaccharides WTPS-1 3 3600~3200 3200~2800 2000~1400 1200~1000 cm 1 4 3430 cm -1, ; 2945 cm 1 C-H ; 1615 cm 1 ; 1328 cm 1 C-N, ; 1153 cm 1 C-O ; 1418 cm 1 =CH 2 ; 1099 cm 1 1010 cm 1 ; 887 cm 1 β-d- β- C-H ; 773 cm 1 D- C-O-C, 887 cm 1, 844 cm 1
3, : 1301 Fig. 3 3 WTPS-1 IR spectrum of Oolong tea polysaccharides WTPS-1, β- ; 810 cm 1, WTPS-1 β- 3.3 乌龙茶 WTPS-1 多糖的单糖组成 - WTPS-1 71.9% WTPS-1, GC-MS, 4 : 1- Rha(16.453 min) 2- Ara(17.297 min) 3- Xyl(17.980 min) 4- Man(27.490 min) 5- Glu(28.005 min) 6- Gal(29.119 min) WTPS-1, WTPS-1, 2.57:3.67:0.57:1:2.16:9.65 3.4 乌龙茶多糖清除 DPPH 自由基 0.2 mg/ml Sevag DEAE WTPS-1, DPPH 91.6% 60.7% 9.20%, DPPH WTPS-1 DPPH 4 (a) WTPS-1 (b) GC-MS 1- (Rha); 2- (Ara); 3- (Xyl); 4- (Man); 5- (Glu); 6- (Gal) Fig. 4 GC-MS total ion chromatograms of acetylated aldononitriles of standard monosaccharides and WTPS-1 polysaccharides hydrolysate 1-Rha; 2- Ara; 3-Xyl; 4-Man; 5-Glu; 6-Gal 4 讨论 10,, WTPS-1
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