原桃胶粗多糖单糖组成分析及其活性研究
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摘要
采用热水提取、乙醇沉淀的方法获得原桃胶粗多糖,并用蒽酮-硫酸法测定该粗多糖中糖含量为93.28%,考马斯亮蓝法测定其蛋白质含量为0.10%。原桃胶粗多糖在2 mol/L硫酸中完全水解后测定其单糖组成,建立了HPLC-ELSD测定原桃胶粗多糖中单糖组成方法。色谱柱:Waters XBridge~(TM) Amide,柱温30℃,进样量10μL,流动相乙腈-水(82:18),流速0.8 mL/min,漂移管110℃,气体流量2.5 L/min。实验证明:该方法精密度、重复性、稳定性、加样回收率良好。测定结果显示,该粗多糖由木糖、阿拉伯糖、甘露糖、葡萄糖、半乳糖组成,其摩尔比为13.97:55.60:1.37:11.94:17.12。同时原桃胶多糖具有较好的抗氧化活性和促巨噬细胞增殖能力。自由基清除实验中原桃胶粗多糖对ABTS~+、OH~—自由基均有一定的清除能力,在6.7 mg/mL时对ABTS~+自由基的清除率达50%,13.7 mg/mL时对OH~—自由基的清除率达50%。在对巨噬细胞的增殖影响中,当多糖浓度为50μg/mL时,与空白组比较,其促进巨噬细胞生长的增殖率达67.16%,能起到增强免疫力的作用。
The crude polysaccharide was obtained by hot water extraction with ethanol precipitation. The content of polysaccharide was 93.28% by anthrone-sulfuric acid method, and the content of protein was 0.10% by Coomassie brilliant blue method. The monosaccharide composition was determined after complete hydrolysis in 2 mol/L sulfuric acid. A HPLC-ELSD method was established to determine the monosaccharide composition in the crude polysaccharide. Chromatogram column: Waters XBridge~(TM) Amide, column temperature 30 ℃, sample injection amount 10 μL, acetonitrile-water(82:18), flow rate 0.8 mL/min, drift tube temperature 110 ℃, gas flow rate 2.5 L/min. The results showed that the crude polysaccharide was composed of xylose, arabinose, mannose, glucose and galactose. The molar ratio was 13.97:55.60:1.37:11.94:17.12. Meanwhile, in the free radical scavenging experiment, the crude polysaccharide of the original peach gum has a good antioxidant activity and an outstanding ability to promote macrophages proliferation. In addition, it has a certain scavenging capacity for ABTS~+ and OH~—free radicals. The clearance rate of ABTS~+ free radicals at 6.7 mg/mL and hydroxyl radicals at 13.7 mg/mL were up to 50%. In the effect on the proliferation of macrophages, compared with the blank group, the proliferation rate was up to 67.16%, when the polysaccharide concentration is 50 μg/mL, which can enhance immunity.
The crude polysaccharide was obtained by hot water extraction with ethanol precipitation. The content of polysaccharide was 93.28% by anthrone-sulfuric acid method, and the content of protein was 0.10% by Coomassie brilliant blue method. The monosaccharide composition was determined after complete hydrolysis in 2 mol/L sulfuric acid. A HPLC-ELSD method was established to determine the monosaccharide composition in the crude polysaccharide. Chromatogram column: Waters XBridge~(TM) Amide, column temperature 30 ℃, sample injection amount 10 μL, acetonitrile-water(82:18), flow rate 0.8 mL/min, drift tube temperature 110 ℃, gas flow rate 2.5 L/min. The results showed that the crude polysaccharide was composed of xylose, arabinose, mannose, glucose and galactose. The molar ratio was 13.97:55.60:1.37:11.94:17.12. Meanwhile, in the free radical scavenging experiment, the crude polysaccharide of the original peach gum has a good antioxidant activity and an outstanding ability to promote macrophages proliferation. In addition, it has a certain scavenging capacity for ABTS~+ and OH~—free radicals. The clearance rate of ABTS~+ free radicals at 6.7 mg/mL and hydroxyl radicals at 13.7 mg/mL were up to 50%. In the effect on the proliferation of macrophages, compared with the blank group, the proliferation rate was up to 67.16%, when the polysaccharide concentration is 50 μg/mL, which can enhance immunity.
引文
[1]《中华本草》编委会.中华本草(第10卷)[M].上海:上海科学技术出版社,1998:86.
[2]姚可成.食物本草[M].北京:人民卫生出版社,1994:441,444.
[3]Wang Y,Lin D,Wang X,et al.The impact of a novel peach gum-derived polysaccharide on postprandial blood glucose control in streptozotocin-induced diabetic mice[J].Int J Biol Macromol,2017,98:379-386.
[4]Yao XC,Cao Y,Wu SJ.Antioxidant activity and antibacterial activity of peach gum derived oligosaccharides[J].Int J Biol Macromol,2013,62:1-3.
[5]郑依玲,董鹏鹏,梅全喜.桃胶特性化学成分药理作用及临床应用研究进展[J].时珍国医国药,2017,28(7):1728-1730.
[6]王飞,丁婷,谷豪.桃胶多糖降糖机制研究[J].中医药导报,2012,18(8):85-88.
[7]吕佳杰,蔡景竹,赵轶凡.桃胶枸杞新型营养果冻的研制[J].安徽农业科学,2016,44(11):86-88.
[8]巢莹,吴平,邓斌,等.天然桃胶软糖的研制[J].安徽农业科学,2015,43(30):240-241,276.
[9]沈洁,周光明,于璐.离子色谱-脉冲安培检测法测定原桃胶中的多糖含量[J].食品工业科技,2017,37(05):298-307.
[10]冯纪南,雷华平,杨德俊,等.电感耦合等离子体原子发射光谱测定桃胶的灰分元素[J].湘南学院学报,2013,37(5):17-19.
[11]董碧莲,吕莉,朱盛山,等.桃胶加工工艺研究概况[J].广东药科大学学报,2018,34(03):403-406.
[12]汪兴杰,张予林,刘吉彬.基于氨基柱的高效液相色谱-示差折光检测器方法分析媚丽葡萄香气糖苷的糖基组成[J].分析化学,2017,45(5):681-686.
[13]李德旺,邓芳,汪胜.蒽酮-硫酸显色法测定栽培白及的白及多糖含量[J].中国药业,2018,27(7):5-7.
[14]杨静华.考马斯亮蓝法测定苦荞麦中可溶性蛋白的含量[J].山西医药杂志,2018,47(2):206-207.
[15]丁成丽,刘力,王强.柱前衍生高效液相色谱法分析新疆桃胶的多糖组成[J].分析科学学报,2012,28(3):319-322.
[16]胡坪,乔晚芳,沈骏文.麦冬多糖单糖组成的分析方法研究[J].药物分析杂志,2013,33(1):50-56.
[17]Han N,Wang L,Song Z,et al.Optimization and antioxidant activity of polysaccharides from Plantago depressa[J].Int J Biol Macromol,2016,93(Pt A):644-654.
[18]Kardosova A,Machova E.Antioxidant activity of medicinal plant polysaccharides[J].Fitoterapia,2006,77(5):367-373.
[19]蔡延渠,邓剑壕,朱志东,等.不同产地原桃胶的多糖含量研究[J].广东药学院学报,2018,34(1):25-28.
[20]王春雨,郭衍俊,张乃强,等.桃胶的制备及性质研究[J].泰山医学院学报,2018,39(1):44-46.
[2]姚可成.食物本草[M].北京:人民卫生出版社,1994:441,444.
[3]Wang Y,Lin D,Wang X,et al.The impact of a novel peach gum-derived polysaccharide on postprandial blood glucose control in streptozotocin-induced diabetic mice[J].Int J Biol Macromol,2017,98:379-386.
[4]Yao XC,Cao Y,Wu SJ.Antioxidant activity and antibacterial activity of peach gum derived oligosaccharides[J].Int J Biol Macromol,2013,62:1-3.
[5]郑依玲,董鹏鹏,梅全喜.桃胶特性化学成分药理作用及临床应用研究进展[J].时珍国医国药,2017,28(7):1728-1730.
[6]王飞,丁婷,谷豪.桃胶多糖降糖机制研究[J].中医药导报,2012,18(8):85-88.
[7]吕佳杰,蔡景竹,赵轶凡.桃胶枸杞新型营养果冻的研制[J].安徽农业科学,2016,44(11):86-88.
[8]巢莹,吴平,邓斌,等.天然桃胶软糖的研制[J].安徽农业科学,2015,43(30):240-241,276.
[9]沈洁,周光明,于璐.离子色谱-脉冲安培检测法测定原桃胶中的多糖含量[J].食品工业科技,2017,37(05):298-307.
[10]冯纪南,雷华平,杨德俊,等.电感耦合等离子体原子发射光谱测定桃胶的灰分元素[J].湘南学院学报,2013,37(5):17-19.
[11]董碧莲,吕莉,朱盛山,等.桃胶加工工艺研究概况[J].广东药科大学学报,2018,34(03):403-406.
[12]汪兴杰,张予林,刘吉彬.基于氨基柱的高效液相色谱-示差折光检测器方法分析媚丽葡萄香气糖苷的糖基组成[J].分析化学,2017,45(5):681-686.
[13]李德旺,邓芳,汪胜.蒽酮-硫酸显色法测定栽培白及的白及多糖含量[J].中国药业,2018,27(7):5-7.
[14]杨静华.考马斯亮蓝法测定苦荞麦中可溶性蛋白的含量[J].山西医药杂志,2018,47(2):206-207.
[15]丁成丽,刘力,王强.柱前衍生高效液相色谱法分析新疆桃胶的多糖组成[J].分析科学学报,2012,28(3):319-322.
[16]胡坪,乔晚芳,沈骏文.麦冬多糖单糖组成的分析方法研究[J].药物分析杂志,2013,33(1):50-56.
[17]Han N,Wang L,Song Z,et al.Optimization and antioxidant activity of polysaccharides from Plantago depressa[J].Int J Biol Macromol,2016,93(Pt A):644-654.
[18]Kardosova A,Machova E.Antioxidant activity of medicinal plant polysaccharides[J].Fitoterapia,2006,77(5):367-373.
[19]蔡延渠,邓剑壕,朱志东,等.不同产地原桃胶的多糖含量研究[J].广东药学院学报,2018,34(1):25-28.
[20]王春雨,郭衍俊,张乃强,等.桃胶的制备及性质研究[J].泰山医学院学报,2018,39(1):44-46.