金银花多糖分子量及单糖组成研究
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摘要
目的探讨金银花分离纯化的多糖组分LTP1、LTP2、LTP3、LTP4的分子量及单糖组成。方法通过热水提取、乙醇沉淀获得金银花粗多糖,经DEAE-52离子交换层析分离纯化,获得金银花多糖LTP1、LTP2、LTP3、LTP4,采用高效凝胶渗透色谱法测定分子量,气相色谱法测定其单糖组成。结果 LTP1重均分子量(Mw)为2016,由L-Rha、D-Ara、L-Fuc、D-Man、D-Glc和D-Gal组成,其摩尔比为1.68∶1.00∶4.42∶13.36∶7.34∶5.07。LTP2 Mw为3907,由L-Rha、D-Ara、L-Fuc、D-Man、D-Glc和D-Gal组成,其摩尔比为2.54∶6.28∶1.00∶5.38∶10.74∶10.44。LTP3 Mw为3039,由L-Rha、D-Ara、D-Glc和D-Gal组成,其摩尔比为1.00∶8.75∶2.59∶2.87。LTP4 Mw为7789,由L-Rha、D-Ara、D-Glc、D-Gal组成,其摩尔比为3.09∶1.86∶5.72∶1.00。结论本研究通过DEAE-52纤维素柱层析分离得到了纯度较好的金银花多糖组分,并确定了其单糖组成,可为金银花多糖的深入研究奠定基础。
Objective To investigate the molecular weight and monosaccharide composition of LTP1, LTP2, LTP3 and LTP4 which were isolated and purified from Lonicerae Japonicae Flos. Methods Polysaccharide from Lonicerae Japonicae Flos was extracted by water extraction and alcohol precipitation, and purified by DEAE-52 ion-exchange column chromatography. The molecular weight and monosaccharide composition of LTP1, LTP2, LTP3 and LTP4 were determined by high performance gel permeation chromatography(HPGPC) and gas chromatography(GC). Results The Mw of LTP1 was 2016, which was composed of L-Rha, D-Ara, L-Fuc, D-Man, D-Glc and D-Gal in a molar ratio of 1.68:1.00:4.42:13.36:7.34:5.07. The Mw of LTP2 was 3907, which was composed of L-Rha, D-Ara, L-Fuc, D-Man, D-Glc and D-Gal in a molar ratio of 2.54:6.28:1.00:5.38:10.74:10.44. The Mw of LTP3 was 3039, which was composed of L-Rha, D-Ara, D-Glc and D-Gal in a molar ratio of 1.00:8.75:2.59:2.87. The Mw of LTP3 was 7789, which was composed of L-Rha, D-Ara, D-Glc and D-Gal in a molar ratio of 3.09:1.86:5.72:1.00. Conclusion The pure polysaccharide is obtained by DEAE-52 cellulose column, and its monosaccharide composition is determined, which has laid the foundation for further research on polysaccharide from Lonicerae Japonicae Flos.
Objective To investigate the molecular weight and monosaccharide composition of LTP1, LTP2, LTP3 and LTP4 which were isolated and purified from Lonicerae Japonicae Flos. Methods Polysaccharide from Lonicerae Japonicae Flos was extracted by water extraction and alcohol precipitation, and purified by DEAE-52 ion-exchange column chromatography. The molecular weight and monosaccharide composition of LTP1, LTP2, LTP3 and LTP4 were determined by high performance gel permeation chromatography(HPGPC) and gas chromatography(GC). Results The Mw of LTP1 was 2016, which was composed of L-Rha, D-Ara, L-Fuc, D-Man, D-Glc and D-Gal in a molar ratio of 1.68:1.00:4.42:13.36:7.34:5.07. The Mw of LTP2 was 3907, which was composed of L-Rha, D-Ara, L-Fuc, D-Man, D-Glc and D-Gal in a molar ratio of 2.54:6.28:1.00:5.38:10.74:10.44. The Mw of LTP3 was 3039, which was composed of L-Rha, D-Ara, D-Glc and D-Gal in a molar ratio of 1.00:8.75:2.59:2.87. The Mw of LTP3 was 7789, which was composed of L-Rha, D-Ara, D-Glc and D-Gal in a molar ratio of 3.09:1.86:5.72:1.00. Conclusion The pure polysaccharide is obtained by DEAE-52 cellulose column, and its monosaccharide composition is determined, which has laid the foundation for further research on polysaccharide from Lonicerae Japonicae Flos.
引文
[1]宋亚玲,倪付勇,赵祎武,等.金银花化学成分研究进展[J].中草药,2014,45(24):3656-3664.
[2]谭政委,夏伟,余永亮,等.金银花化学成分及其药理学研究进展[J].安徽农业科学,2018,46(9):26-28,123.
[3]刘蓓,刘玉红.金银花多糖对脾淋巴细胞的增殖作用[J].中国实用医药,2013,8(11):244-245.
[4]刘玉国,刘玉红,蒋海强.金银花多糖对小鼠S180肉瘤的抑制作用与机制研究[J].肿瘤学杂志,2012,18(8):584-587.
[5]贾伟,毛淑敏,张盼盼,等.金银花多糖体内抗病毒作用研究[J].辽宁中医药大学学报,2018,20(6):1-4.
[6]王露懿,朱华旭.多糖资源的开发应用现状及存在问题分析[J].药物生物技术,2018,25(1):90-94.
[7]贾伟,毛淑敏,金娟,等.薄层色谱法联合气相色谱法分析金银花多糖的单糖组分研究[J].山东中医杂志,2017,36(4):326-328.
[8]毛淑敏,许家珍,周帅飞,等.苯酚-硫酸法联合DNS法测定金银花不同花期多糖的含量[J].中南药学,2015,13(1):65-67.
[9]李启艳,胡德福,张雪梅,等.党参多糖提取纯化工艺优化及其组成研究[J].中草药,2016,47(15):2663-2667.
[10]杨树娟,姜文圣,钱如贵,等.HPGPC法测定库拉索芦荟多糖的分子量和含量[J].今日药学,2016,26(11):786-791.
[11]穆俊,邓旭坤,江善青,等.密蒙花多糖提取工艺的正交实验法优选及其单糖组成的气相色谱法分析[J].时珍国医国药,2017,28(3):538-540.
[12]赵鹏,张婷婷.响应面法优化金银花多糖梭甲基化工艺及抗氧化性研究[J].天然产物研究与开发,2015,27(1):114-119,190.
[13]李尔春.金银花多糖的分离纯化与生物活性研究[D].西安:陕西师范大学,2008.
[14]邓庆华.用正交试验法优化金银花多糖提取工艺[D].长春:东北师范大学,2008.
[15]秦建鲜,黄锁义.中药多糖相对分子质量测定方法的概述[J].中国实验方剂学杂志,2015,21(14):226-229.
[16]刘玉明,李家春,钱甜甜,等.方格星虫多糖分子量的高效凝胶渗透色谱法测定[J].时珍国医国药,2013,24(12):2857-2858.
[17]侯重文,朱梓昂,张金华,等.高效凝胶色谱法测定贻贝多糖含量及分子量[J].中国生化药物杂志,2016,36(11):9-12.
[2]谭政委,夏伟,余永亮,等.金银花化学成分及其药理学研究进展[J].安徽农业科学,2018,46(9):26-28,123.
[3]刘蓓,刘玉红.金银花多糖对脾淋巴细胞的增殖作用[J].中国实用医药,2013,8(11):244-245.
[4]刘玉国,刘玉红,蒋海强.金银花多糖对小鼠S180肉瘤的抑制作用与机制研究[J].肿瘤学杂志,2012,18(8):584-587.
[5]贾伟,毛淑敏,张盼盼,等.金银花多糖体内抗病毒作用研究[J].辽宁中医药大学学报,2018,20(6):1-4.
[6]王露懿,朱华旭.多糖资源的开发应用现状及存在问题分析[J].药物生物技术,2018,25(1):90-94.
[7]贾伟,毛淑敏,金娟,等.薄层色谱法联合气相色谱法分析金银花多糖的单糖组分研究[J].山东中医杂志,2017,36(4):326-328.
[8]毛淑敏,许家珍,周帅飞,等.苯酚-硫酸法联合DNS法测定金银花不同花期多糖的含量[J].中南药学,2015,13(1):65-67.
[9]李启艳,胡德福,张雪梅,等.党参多糖提取纯化工艺优化及其组成研究[J].中草药,2016,47(15):2663-2667.
[10]杨树娟,姜文圣,钱如贵,等.HPGPC法测定库拉索芦荟多糖的分子量和含量[J].今日药学,2016,26(11):786-791.
[11]穆俊,邓旭坤,江善青,等.密蒙花多糖提取工艺的正交实验法优选及其单糖组成的气相色谱法分析[J].时珍国医国药,2017,28(3):538-540.
[12]赵鹏,张婷婷.响应面法优化金银花多糖梭甲基化工艺及抗氧化性研究[J].天然产物研究与开发,2015,27(1):114-119,190.
[13]李尔春.金银花多糖的分离纯化与生物活性研究[D].西安:陕西师范大学,2008.
[14]邓庆华.用正交试验法优化金银花多糖提取工艺[D].长春:东北师范大学,2008.
[15]秦建鲜,黄锁义.中药多糖相对分子质量测定方法的概述[J].中国实验方剂学杂志,2015,21(14):226-229.
[16]刘玉明,李家春,钱甜甜,等.方格星虫多糖分子量的高效凝胶渗透色谱法测定[J].时珍国医国药,2013,24(12):2857-2858.
[17]侯重文,朱梓昂,张金华,等.高效凝胶色谱法测定贻贝多糖含量及分子量[J].中国生化药物杂志,2016,36(11):9-12.