月季果实多糖的纯化及部分特性研究

Xuejiao Zhang, Chenzhong Jin, Yihong Hu, Yunyun Zhou, Wang Shuanghui
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引用次数: 0

摘要

对中药月桂多糖的提取优化、纯化及部分特性进行了研究。蔷薇属蔷薇科,其果实在中国、日本等亚洲国家被广泛用作保健食品[1]。本研究采用响应面法优化了枸杞子水溶性多糖的提取工艺。采用中心组合设计优化提取工艺参数。最佳提取条件为:提取温度95℃,提取时间2.5 h,水料比22:1,提取次数3次。在最佳条件下,实验产率为9.55±0.1%,与预测产率吻合较好。采用二乙胺乙醇-Sepharose柱层析法和Sepharose CL-4B柱层析法分别对果实粗多糖进行纯化,得到了6个主要部分(RLP-I-1、RLP-I-2、RLP-I-3、RLP-II-1、RLP-II-2和RLP-II-3)。主要组分的分子量采用高效凝胶渗透色谱法测定,采用差示折射率检测器(Waters2410,美国)和G5000 PWxl柱(7.8 mm×300 mm, TOSOH,日本)串联G3000 PWxl柱(7.8 mm×300 mm, TOSOH,日本)。RLP-I-1、RLP-I-2、RLP-I-3、RLP-II-1、RLP-II-2、RLP-II-3的平均分子量分别为8930 Da、9498 Da、9662 Da、7673 Da、8558 Da和8323 Da。6种多糖的傅里叶变换红外光谱相同,-OH基团在3400 cm 1处出现宽的拉伸强度特征峰,而在2935 ~ 2939 cm处出现弱的C-H拉伸带。1616 cm处的峰来自于-OH基团的弯曲振动吸收。1460 ~ 1200cm范围内的吸收峰为碳-氢的变角振动。1285 ~ 1020 cm范围内的峰值对应于C-O拉伸振动。880 cm左右的吸收带是由α型糖苷键形成的。峰分布在3600 ~ 3200 cm和1655 ~ 1615 cm,表明这些化合物为碳水化合物。此外,在1715 cm附近没有检测到峰,表明果聚糖不含糖醛酸[2]。在本研究中,用清除率来评价6种多糖对超氧阴离子、羟基和1,1 -二苯基-2-苦味酰肼(DPPH)自由基的体外抗氧化活性。结果表明,RLP-II-1、RLP-II-2和RLP-II-3对超氧阴离子、羟基和DPPH自由基具有较强的清除活性。结果表明,响应面法是一种有效的提取枸杞多糖的方法,可开发枸杞多糖作为功能性食品的抗氧化剂。
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Purification and Partial Characteristics of Polysaccharides from Rosa Laevigata Michx Fruits
Extended Abstract Extraction optimization, purification and partial characteristics of polysaccharides from the traditional Chinese herb Rosae Laevigatae Michx were investigated. R. Leavigata belongs to the Rosa genus and Rosaceae family, and its fruits are widely used as health food in China, Japan and the other Asian countries [1]. In this study, the response surface methodology was used to optimize the extraction conditions of water-soluble polysaccharides from R. Laevigatae fruits. The central composite design was used to optimize the extraction processing parameters. The optimum extraction conditions were extraction temperature 95C, extraction time 2.5 h, water to raw material ratio 22:1, and extraction frequency 3. Under the optimum conditions, the experimental yield was 9.55±0.1%, which is in good agreement with the predicted yield. Six major fractions (RLP-I-1, RLP-I-2, RLP-I-3, RLP-II-1, RLP-II-2 and RLP-II-3) were successfully obtained by purifying the crude polysaccharides extracted from the fruits by using diethylaminoethanol-Sepharose column chromatography and Sepharose CL-4B column chromatography respectively. The molecular weights of major fractions were determined by the high performance gel permeation chromatography with a differential refractive index detector (Waters2410, USA) and a G5000 PWxl column (7.8 mm×300 mm, TOSOH, Japan) connected in series with a G3000 PWxl column (7.8 mm×300 mm, TOSOH, Japan). The average molecular weight of the RLP-I-1, RLP-I-2, RLP-I-3, RLP-II-1, RLP-II-2, RLP-II-3 were estimated to be 8930 Da, 9498 Da , 9662 Da, 7673 Da, 8558 Da and 8323 Da, respectively. The fourier transform infrared spectra of the six polysaccharides were identical, and the broad stretching intense characteristic peak was shown at 3400 cm 1 for the -OH group, whereas a weak C-H stretching band was observed from 2935-2939 cm. The peaks at 1616 cm were from the bending vibration absorption of -OH group. Absorption peaks ranged from 1460-1200 cm were the variable angle vibrations of C-H. The peaks from 1285 cm to 1020 cm corresponded to C-O stretching vibrations. The absorption band round 880 cm was due to the α-type glycosidic bond. Peaks ranged from 3600-3200 cm and 1655-1615 cm revealed that these compounds were carbohydrates. Moreover, no peaks were detected near 1715 cm suggesting that the fructan had no uronic acid [2]. In the present study, the anti-oxidative activities of the six polysaccharides toward super-oxide anion, hydroxyl, and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals in vitro were evaluated in terms of scavenging rate. The assays suggested that RLP-II-1, RLP-II-2 and RLP-II-3 presented higher scavenging activity toward superoxide anions, hydroxyl and DPPH radicals. The results indicated that the response surface methodology was an effective method for the extraction of polysaccharides from the R. Laevigatae fruits, and the polysaccharides could be explored as a potential antioxidant agent for use in functional foods.
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