枣中功能性糖的研究
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摘要
枣(Ziziphus jujuba Mill.)隶属于鼠李科枣属,主要分布于亚洲和北美洲的亚热带和热带地区。枣果中营养成分丰富,主要包含糖、多酚、环磷酸腺苷、环磷酸鸟苷、三萜及其苷类、生物碱、矿质元素、维生素及其它活性物质,是传统的药食兼用佳品。对枣果中的化学成分研究远不及同类的传统中药,糖作为枣果中含量最大的组分,在其制备、测定及生物学活性方面的研究仍不够深入,为了明确果中主要化学成分,确定枣中糖类成分的组成及生物学活性,以利于今后对枣的合理开发利用,本研究对二十种具有代表性的枣中主要营养成分进行分析,选择其中栽培范围较广的金丝小枣为研究目标,对其多糖、低聚糖和单糖的制备及组成进行研究,并对低聚糖进行双歧杆菌体外增殖实验。此外,还将金丝小枣与其它七种枣中水溶性多糖的单糖组成及其摩尔比进行了比较。
从国家红枣良种繁育基地选取具有代表性的二十个品种,对其进行总糖、还原糖、多糖、脂肪、蛋白质、矿质元素、Vc等10项营养指标测定。实验结果表明:鲜食型品种、制干型品种和兼用型品种在水分、还原糖、粗纤维和Ca含量上存在差异较大。
金丝小枣是河北省的优势栽培品种,以其为研究对象,通过水提醇沉得到枣多糖和小分子糖两种组分。以Sevage法和反复醇洗法除去多糖中的蛋白质和色素,干燥后备用。小分子糖通过超滤与纳滤相结合的方式进行分级,得到500-1000Da之间的低聚糖组分以及小于500D的单糖组分。
建立了柱前衍生化HPLC法测定枣多糖中单糖组成的方法,以1-苯基-3-甲基-5-吡唑啉酮为衍生剂,20mmol·L~(-1)磷酸盐缓冲液(pH 6.7)-乙腈(83∶17)为流动相。枣多糖用三氟乙酸水解后与1-苯基-3-甲基-5-吡唑啉酮反应,生成具有紫外吸收的物质,对其进行HPLC分析,确定金丝小枣中水溶性多糖的单糖组成及其摩尔比,并将金丝小枣与赞皇大枣、梨枣、宜城圆枣、无核小枣、晋枣、骏枣、大荔龙枣七个品种的枣中水溶性多糖进行比较。试验结果表明:八种枣多糖中均含有葡萄糖、阿拉伯糖、半乳糖、木糖、甘露糖、半乳糖醛酸六种单糖,但各种枣多糖中单糖组成比例上存在很大差异,甘露糖在各多糖中所占比例均为最小,金丝小枣中半乳糖醛酸所占比例最大,其鼠李糖、半乳糖醛酸以及阿拉伯糖的含量均高于另外七种枣。
建立了离子色谱方法,将得到的500-1000Da范围和小于500Da范围的两个组分分别进行离子色谱分析。结果表明:低聚糖主要包含5个均一性组分构成,其聚合度约为3-7。分子量小于500Da的组分由葡萄糖、果糖和蔗糖组成,其构成比例为Glu: Suc: Fru= 4.48: 0.32: 4.66。
通过双歧杆菌体外增殖试验对分离得到的金丝小枣低聚糖的活性进行研究。结果表明:低聚糖对动物双歧杆菌体外生长具有促进作用。基础培养基中分别添加0.1%、0.5%、1%的低聚糖对双歧杆菌体外生长均具有促进作用,当低聚糖添加浓度为0.5%时,促进生长效果效果最明显。
本研究选取具有代表性的二十种枣作为研究对象,对其基本营养成分进行分析比较;建立了测定枣多糖中单糖组成的高效液相色谱方法,对金丝小枣中多糖进行分析,并将其与其它栽培范围较广的七种枣中多糖进行比较,研究发现:不同品种的枣多糖中单糖在组成比例上存在很大差异,由此推断其生物学活性上可能也存在一定差异;采用膜分离方式得到金丝小枣低聚糖,首次采用离子色谱法对其组成进行了分析,并通过双歧杆菌体外促生长实验,验证了金丝小枣低聚糖对双歧杆菌体外生长具有促进作用。为枣的综合利用及功能性糖分进一步研究提供了科学依据。
Chinese jujube (Ziziphus jujuba Mill.) belongs to the Ziziphus mill.(Rhamnaceae), which is mainly distributed in the subtropical and tropical regions of Asia and America. Date-the fruit of jujube can serves as both traditional Chinese medicine and invigorant since date is rich in nutrients, including sugars, polyphenols, cyclic adenosine monophosphate (cAMP), cyclic Ukraine monophosphate (cGMP), triterpenoids and their glycosides, alkaloids, flavonides, Phenitic acids inorganic elements, vitamins and other nutritional active ingredients. But the chemical components research of date, compared with the other traditional Chinese medicines, is still at a lower level. In order to develop and utilize the date reasonably, this study analyzed the major nutrients of 20 kinds of representative dates to identify the chemical components in jujube, determine the composition and biological activity of water-soluble sugar. Taking the Zizyphus jujuba cv. Jinsixiaozao, which is a widely cultivated kind of date, as the research objective, this research investigate the preparation and composition of polysaccharides, oligosaccharides and monosaccharides as well as the vitro experiments of Bifidobacterium Proliferation. In addition, this research makes a comparison between Zizyphus jujuba cv. Jinsixiaozao and the other seven varieties of jujube in the monosaccharide composition and its molar ratio of the water-soluble polysaccharides.
The research select twenty representative varieties from National jujube seed breeding base to measure the ten nutrition indicators including total sugars, reducing sugars, polysaccharides, fats, proteins, mineral elements, Vc and the others. The result shows that fresh varieties, dried varieties and compatible varieties are mainly different in water, reducing sugar fiber and Calcium content.
Taking Zizyphus jujuba cv. Jinsixiaozao which is an advantage cultivar of Hebei province as the study object, the researchers extract polysaccharide and Small molecular sugar through Alcohol sedimenting technique. The water-soluble polysaccharides were obtained by ethanol precipitation, use Sevage method and alcohol repeated method remove protein and depigment in polysaccharides. Using a combination of ultrafiltration and nanofiltration obtain Oligosaccharides and monosaccharide.
Establish a high performance liquid chromatographic method of pre-column derivatization for analysis monosaccharide composition of water-soluble polysaccharides from jujube. The separation was carried out on a RP-C18 column using pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) and UV detection at 250 nm with a mobile phase consist of 0.02M phosphate buffer (PH=6.7) and acetonitrile in the ratio of 82:18. Polysaccharides hydrolysis by trifluoroacetic acid and derivate by PMP, Application of the HPLC method determined the monosaccharide composition and their molar ratio of polysaccharides from Zyzysphys jujuba cv. Jinxixiaozao. Than compared with other seven common cultivars. The result shows that each of the sample consisted of mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose, but monosaccharide composition in deferent molar ratio. The proportion of Mannose is the smallest in each of the sample. Zizyphus jujuba cv. jinsixiaozao content galacturonic acid with the largest proportion, rhamnose, galacturonic acid and arabinose which content in Zizyphus jujuba cv. Jinsixiaozao are higher than other seven cultivars of jujube.
Establish ion chromatography method to analysis each component classification by membrane was analysis by ion chromatography, respectively. The results showed that oligosaccharides distribution 500D-1000D membrane retention range, there are five main components constitute. Degree of polymerization is about three to seven. The component with molecular weight less than the 500D is composed of glucose, sucrose and fructose in ratio of 4.48: 0.32: 4.66.
The biological activity of oligosaccharides from Zizyphus jujuba cv. Jinsixiaozao was determined by growth promoting effect on bifidobacteria in Vitro. The results showed that oligosaccharides in Zizyphus jujuba cv. Jinsixiaozao can promote Bifidobacterium Animals growth in vitro. By adding 0.1%, 0.5%, 1.0% of oligosaccharides in the basal medium are all have promoting growth effect to Bifidobacterium Animals growth in vitro. When the concentration of adding oligosaccharides is 0.5%, the promote effect is the best.
Selecting twenty representative varieties of Jujube as research object, this research makes an analysis and comparison of their basic nutrients; establish a high performance liquid chromatography method to determinate of the monosaccharide composition of polysaccharides. After analyzing to Polysaccharides in Zizyphus jujuba cv. Jinsixiaozao and comparing to the other seven widely cultivated jujubes, the research result shows that monosaccharide composition of each polysaccharide in deferent molar ratio. Therefore, the biological activity of different varieties of jujube polysaccharides may have large difference. Oligosaccharides purified by membrane. For the first time, the composition of oligosaccharide in Jujube was determined by ion chromatography and its biological activity was measured by microbiology experiment. The experiment verifies the significance of oligosaccharides on promote the Bifidobacterium growth in vitro. This research provides scientific basis to further study of date’s Comprehensive use and functional sugar analysis.
从国家红枣良种繁育基地选取具有代表性的二十个品种,对其进行总糖、还原糖、多糖、脂肪、蛋白质、矿质元素、Vc等10项营养指标测定。实验结果表明:鲜食型品种、制干型品种和兼用型品种在水分、还原糖、粗纤维和Ca含量上存在差异较大。
金丝小枣是河北省的优势栽培品种,以其为研究对象,通过水提醇沉得到枣多糖和小分子糖两种组分。以Sevage法和反复醇洗法除去多糖中的蛋白质和色素,干燥后备用。小分子糖通过超滤与纳滤相结合的方式进行分级,得到500-1000Da之间的低聚糖组分以及小于500D的单糖组分。
建立了柱前衍生化HPLC法测定枣多糖中单糖组成的方法,以1-苯基-3-甲基-5-吡唑啉酮为衍生剂,20mmol·L~(-1)磷酸盐缓冲液(pH 6.7)-乙腈(83∶17)为流动相。枣多糖用三氟乙酸水解后与1-苯基-3-甲基-5-吡唑啉酮反应,生成具有紫外吸收的物质,对其进行HPLC分析,确定金丝小枣中水溶性多糖的单糖组成及其摩尔比,并将金丝小枣与赞皇大枣、梨枣、宜城圆枣、无核小枣、晋枣、骏枣、大荔龙枣七个品种的枣中水溶性多糖进行比较。试验结果表明:八种枣多糖中均含有葡萄糖、阿拉伯糖、半乳糖、木糖、甘露糖、半乳糖醛酸六种单糖,但各种枣多糖中单糖组成比例上存在很大差异,甘露糖在各多糖中所占比例均为最小,金丝小枣中半乳糖醛酸所占比例最大,其鼠李糖、半乳糖醛酸以及阿拉伯糖的含量均高于另外七种枣。
建立了离子色谱方法,将得到的500-1000Da范围和小于500Da范围的两个组分分别进行离子色谱分析。结果表明:低聚糖主要包含5个均一性组分构成,其聚合度约为3-7。分子量小于500Da的组分由葡萄糖、果糖和蔗糖组成,其构成比例为Glu: Suc: Fru= 4.48: 0.32: 4.66。
通过双歧杆菌体外增殖试验对分离得到的金丝小枣低聚糖的活性进行研究。结果表明:低聚糖对动物双歧杆菌体外生长具有促进作用。基础培养基中分别添加0.1%、0.5%、1%的低聚糖对双歧杆菌体外生长均具有促进作用,当低聚糖添加浓度为0.5%时,促进生长效果效果最明显。
本研究选取具有代表性的二十种枣作为研究对象,对其基本营养成分进行分析比较;建立了测定枣多糖中单糖组成的高效液相色谱方法,对金丝小枣中多糖进行分析,并将其与其它栽培范围较广的七种枣中多糖进行比较,研究发现:不同品种的枣多糖中单糖在组成比例上存在很大差异,由此推断其生物学活性上可能也存在一定差异;采用膜分离方式得到金丝小枣低聚糖,首次采用离子色谱法对其组成进行了分析,并通过双歧杆菌体外促生长实验,验证了金丝小枣低聚糖对双歧杆菌体外生长具有促进作用。为枣的综合利用及功能性糖分进一步研究提供了科学依据。
Chinese jujube (Ziziphus jujuba Mill.) belongs to the Ziziphus mill.(Rhamnaceae), which is mainly distributed in the subtropical and tropical regions of Asia and America. Date-the fruit of jujube can serves as both traditional Chinese medicine and invigorant since date is rich in nutrients, including sugars, polyphenols, cyclic adenosine monophosphate (cAMP), cyclic Ukraine monophosphate (cGMP), triterpenoids and their glycosides, alkaloids, flavonides, Phenitic acids inorganic elements, vitamins and other nutritional active ingredients. But the chemical components research of date, compared with the other traditional Chinese medicines, is still at a lower level. In order to develop and utilize the date reasonably, this study analyzed the major nutrients of 20 kinds of representative dates to identify the chemical components in jujube, determine the composition and biological activity of water-soluble sugar. Taking the Zizyphus jujuba cv. Jinsixiaozao, which is a widely cultivated kind of date, as the research objective, this research investigate the preparation and composition of polysaccharides, oligosaccharides and monosaccharides as well as the vitro experiments of Bifidobacterium Proliferation. In addition, this research makes a comparison between Zizyphus jujuba cv. Jinsixiaozao and the other seven varieties of jujube in the monosaccharide composition and its molar ratio of the water-soluble polysaccharides.
The research select twenty representative varieties from National jujube seed breeding base to measure the ten nutrition indicators including total sugars, reducing sugars, polysaccharides, fats, proteins, mineral elements, Vc and the others. The result shows that fresh varieties, dried varieties and compatible varieties are mainly different in water, reducing sugar fiber and Calcium content.
Taking Zizyphus jujuba cv. Jinsixiaozao which is an advantage cultivar of Hebei province as the study object, the researchers extract polysaccharide and Small molecular sugar through Alcohol sedimenting technique. The water-soluble polysaccharides were obtained by ethanol precipitation, use Sevage method and alcohol repeated method remove protein and depigment in polysaccharides. Using a combination of ultrafiltration and nanofiltration obtain Oligosaccharides and monosaccharide.
Establish a high performance liquid chromatographic method of pre-column derivatization for analysis monosaccharide composition of water-soluble polysaccharides from jujube. The separation was carried out on a RP-C18 column using pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP) and UV detection at 250 nm with a mobile phase consist of 0.02M phosphate buffer (PH=6.7) and acetonitrile in the ratio of 82:18. Polysaccharides hydrolysis by trifluoroacetic acid and derivate by PMP, Application of the HPLC method determined the monosaccharide composition and their molar ratio of polysaccharides from Zyzysphys jujuba cv. Jinxixiaozao. Than compared with other seven common cultivars. The result shows that each of the sample consisted of mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose, but monosaccharide composition in deferent molar ratio. The proportion of Mannose is the smallest in each of the sample. Zizyphus jujuba cv. jinsixiaozao content galacturonic acid with the largest proportion, rhamnose, galacturonic acid and arabinose which content in Zizyphus jujuba cv. Jinsixiaozao are higher than other seven cultivars of jujube.
Establish ion chromatography method to analysis each component classification by membrane was analysis by ion chromatography, respectively. The results showed that oligosaccharides distribution 500D-1000D membrane retention range, there are five main components constitute. Degree of polymerization is about three to seven. The component with molecular weight less than the 500D is composed of glucose, sucrose and fructose in ratio of 4.48: 0.32: 4.66.
The biological activity of oligosaccharides from Zizyphus jujuba cv. Jinsixiaozao was determined by growth promoting effect on bifidobacteria in Vitro. The results showed that oligosaccharides in Zizyphus jujuba cv. Jinsixiaozao can promote Bifidobacterium Animals growth in vitro. By adding 0.1%, 0.5%, 1.0% of oligosaccharides in the basal medium are all have promoting growth effect to Bifidobacterium Animals growth in vitro. When the concentration of adding oligosaccharides is 0.5%, the promote effect is the best.
Selecting twenty representative varieties of Jujube as research object, this research makes an analysis and comparison of their basic nutrients; establish a high performance liquid chromatography method to determinate of the monosaccharide composition of polysaccharides. After analyzing to Polysaccharides in Zizyphus jujuba cv. Jinsixiaozao and comparing to the other seven widely cultivated jujubes, the research result shows that monosaccharide composition of each polysaccharide in deferent molar ratio. Therefore, the biological activity of different varieties of jujube polysaccharides may have large difference. Oligosaccharides purified by membrane. For the first time, the composition of oligosaccharide in Jujube was determined by ion chromatography and its biological activity was measured by microbiology experiment. The experiment verifies the significance of oligosaccharides on promote the Bifidobacterium growth in vitro. This research provides scientific basis to further study of date’s Comprehensive use and functional sugar analysis.
引文
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