[1]汪小君,岳盼盼,杨子颖,等.老龙皮多糖的分离纯化及体外活性研究[J].林业工程学报,2019,4(04):71-77.[doi:10.13360/j.issn.2096-1359.2019.04.011]
 WANG Xiaojun,YUE Panpan,YANG Ziying,et al.Isolation, purification and antioxidant activity determination of polysaccharides from Lobariakoor kauae Yoshim[J].Journal of Forestry Engineering,2019,4(04):71-77.[doi:10.13360/j.issn.2096-1359.2019.04.011]
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老龙皮多糖的分离纯化及体外活性研究()
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《林业工程学报》[ISSN:1001-8081/CN:32-1160/S]

卷:
4
期数:
2019年04期
页码:
71-77
栏目:
林产化学加工
出版日期:
2019-07-09

文章信息/Info

Title:
Isolation, purification and antioxidant activity determination of polysaccharides from Lobariakoor kauae Yoshim
文章编号:
2096-1359(2019)04-0071-07
作者:
汪小君岳盼盼杨子颖熊金衫孙玲玲彭锋*
北京林业大学材料科学与技术学院,林木生物质化学北京市重点实验室,北京 100083
Author(s):
WANG Xiaojun YUE Panpan YANG Ziying XIONG Jinshan SUN Lingling PENG Feng*
College of Materials Science and Technology, Beijing Key Laboratory of Forest Biomass Chemistry, Beijing Forestry University, Beijing 100083, China
关键词:
老龙皮 多糖 分级沉淀 结构鉴定 抗氧化活性
Keywords:
Lobariakoor kauae Yoshim polysaccharides fractional precipitation structural characteristics antioxidant activity
分类号:
R285.5
DOI:
10.13360/j.issn.2096-1359.2019.04.011
文献标志码:
A
摘要:
地衣植物老龙皮富含多糖和三萜皂苷类化合物,是一种资源丰富的具有抗炎、抗氧化等活性的中草药。以老龙皮为原料,通过甲醇抽提脱脂、水浸提、三氯乙酸(TCA)法脱蛋白等方式粗提,然后分别采用梯度浓度乙醇分级沉淀法和DEAE-52纤维素柱层析的方法对老龙皮多粗多糖进行分级纯化。梯度浓度乙醇(乙醇质量浓度分别为15%,30%,50%和70%)分级沉淀得到4种多糖组分,以DEAE-52纤维素柱层析法洗脱得到5种老龙皮多糖组分。采用高效阴离子色谱(HPAEC)与红外光谱法(FT-IR)分析了老龙皮多糖组分的单糖组成和结构。研究结果表明,老龙皮多糖的单糖组成主要有阿拉伯糖、半乳糖、葡萄糖、甘露糖和木糖。其中,甘露糖含量最高,其次是半乳糖和葡萄糖。老龙皮多糖的红外光谱特征峰为3 417,2 911,1 659,1 400,1 026和831 cm-1。利用乙醇分级沉淀和DEAE-52纤维素柱层析洗脱所得的9种老龙皮多糖组分进行体外抗氧化活性研究,结果显示老龙皮多糖具有较强的清除DPPH自由基能力和总还原力。其中:50%质量浓度乙醇沉淀的老龙皮多糖组分的清除能力最强,清除率达80.10%; 15%质量浓度乙醇沉淀的老龙皮多糖组分的还原能力最强,其吸光值最高为0.74。
Abstract:
Being rich in polysaccharides and triterpene saponins, Lobariakoor kauae Yoshim is a resource-rich Chinese herbal medicine with anti-inflammatory, anti-oxidant and other medical functions.In this research, the polysaccharides were extracted with water at 80 ℃ from dewaxed Lobariakoor kauae Yoshim.The protein in the obtained polysaccharides was removed by the treatment of trichloroacetic acid.Then the polysaccharides were fractionated by the graded ethanol precipitation and diethyl-aminoethanol(DEAE)-cellulose-52 chromatography, four(P15, P30, P50 and P70)and five(Z1, Z2, Z3, Z4 and Z5)polysaccharide components were obtained, respectively.The sugar composition, structural properties of nine polysaccharide fractions were characterized by the high-performance anion exchange chromatography and Fourier transform infrared spectroscopy(FT-IR).The results showed that the main monosaccharide components were arabinose, galactose, glucose, mannose and xylose.The mannose was the predominant sugar composition, followed by glucose and galactose.The FT-IR results showed that the characteristic polysaccharides peaks were 3 417, 2 911, 1 659, 1 400, 1 026 and 831 cm-1.The infrared spectra of polysaccharides had the same contour and the structure of the polysaccharides were similar.The intensities of peaks were slightly different, which made contents of the acetyl and uronic acid groups slightly different.The antioxidant activities of 9 polysaccharides was obtained by the ethanol fractional precipitation and the DEAE-52 cellulose column chromatography was studied in vitro.The results showed that the polysaccharides had good ability to scavenge 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radicals and total reducing ability.Among them, the polysaccharide fraction precipitated by 50% ethanol had the strongest scavenging DPPH free radicalability, with a scavenging rate of 80.10%.The polysaccharide fractions precipitated by 15% ethanol had the strongest reducing ability, and its absorbance was up to 0.74.In addition, among the five polysaccharides fractionated by column chromatography, the absorbance of Z5 was significantly higher than that of the other four types.It can be seen that Z5 had the strongest antioxidant activity, which was consistent with the results of the DPPH.

参考文献/References:

[1] 中药辞海(第一卷)[M].北京: 中国医药科技出版社, 1993: 1-899.
[2] 杜远东.老龙皮的药效物质及质量标准研究[D].咸阳: 陕西中医学院, 2012.
DU Y D.Study on the pharmacodynamic substances and quality standards of Lobariakoor kauae Yoshim[D].Xianyang: Shaanxi University of Chinese Medicine, 2012.
[3] MACMILLAN J G, BROWNE J L.Aregiospecific synthesis of hematommic acid[J].The Journal of Organic Chemistry, 1977, 42(14): 2526-2528.DOI: 10.1021/jo00434a047.
[4] 李志东.两种地衣植物的化学成分及其抗肿瘤作用研究[D].广州: 广州中医药大学,2015.
LI Z D.Chemical constituents of two lichens and their anti-tumor effects[D].Guangzhou: Guangzhou University of Chinese Medicine, 2015.
[5] 陈晓白, 蒋夏荣, 杨秋元, 等.山银花多糖提取工艺优化及其抗氧化活性研究[J].中国食品添加剂, 2018(11): 155-161.DOI: 10.3969/j.issn.1006-2513.2018.11.019.
CHEN X B, JIANG X R, YANG Q Y, et al.Optimization on extraction process of polysaccharide from Lonicerae flos and its antioxidant activity[J].China Food Additives, 2018(11): 155-161.
[6] 金黎明, 陈苛蒙, 包艳春, 等.核桃楸皮多糖的抗氧化及抗菌作用研究[J].食品科技, 2018, 43(10): 252-255.DOI: CNKI:SUN:SSPJ.0.2018-10-043.
JIN L M, CHEN K M, BAO Y C, et al.Antioxidant and antibacterial activity of polysaccharide from Juglans mandshurica Maxim bark[J].Food Science and Technology, 2018, 43(10): 252-255.
[7] 杨柠, 张赫, 张凤娟, 等.黑木耳多糖对TM3细胞增殖的影响[J].吉林医药学院学报,2019, 40(1): 1-4.DOI: 10.13845/j.cnki.issn1673-2995.2019.01.001.
YANG N, ZHANG H, ZHANG F J, et al.Effects of auricularia auricula polysaccharide on the proliferation of TM3 cells[J].Journal of Jilin Medical University, 2019, 40(1): 1-4.
[8] 杨晓莉, 罗晶, 郭耀武, 等.老龙皮水和乙醇提取部位的抗炎和体外抑菌实验研究[J].陕西中医, 2012, 33: 1434-1435.DOI: 10.3969/j.issn.1000-7369.2012.10.117.
YANG X L, LUO J, GUO Y W, et al.Anti-inflammatory and in vitro antibacterial experiments of Lobariakoor kauae Yoshim extracted by water and ethanol extraction sites[J].Shaanxi Journal of Traditional Chinese Medicine, 2012, 33: 1434-1435.
[9] 王晓梅, 李健, 李宗孝, 等.老龙皮提取物的体外抗氧化活性研究[J].现代中医药, 2010(5): 79-80.DOI: 10.13424/j.cnki.mtcm.2010.05.036.
WANG X M, LI J, LI Z X, et al.In vitro antioxidant activity of Lobariakoor kauae Yoshim[J].Modern Chinese Medicine, 2010(5): 79-80.
[10] 张红, 孙婷婷, 王海峰, 等.老龙皮多糖分离纯化及糖组成研究[J].中国药业, 2016, 25(13): 40-43.
ZHANG H, SUN T T, WANG H F, et al.Separation, purification and sugar composition of Lobariakoor kauae Yoshim[J].Chinese Pharmaceuticals, 2016, 25(13): 40-43.
[11] 朱银玲.杨桃多糖的提取及免疫活性研究[D].广州: 广东工业大学, 2008.
ZHU Y L.Extraction and immunological activity of carambola polysaccharide[D].Guangzhou: Guangdong University of Technology, 2008.
[12] 李知敏, 王伯初, 周菁, 等.植物多糖提取液的几种脱蛋白方法的比较分析[J].重庆大学学报(自然科学版), 2004, 27(8): 57-59.DOI: 10.3969/j.issn.1000-582X.2004.08.014.
LI Z M, WANG B C, ZHOU J, et al.Comparative analysis of several deproteinization methods for plant polysaccharide extracts[J].Journal of Chongqing University(Natural Science), 2004,27(8): 57-59.
[13] 王金鹏, 陈寒青, 邓力, 等.刺五加多糖提取过程中不同脱蛋白方法的比较研究[J].天然产物研究与开发,2009, 21(1): 155-158.DOI: 10.3969/j.issn.1001-6880.2009.01.036.
WANG J P, CHEN H Q, DENG L, et al.Comparative study on different deproteinization methods in the extraction process of Acanthopanax Senticosus polysaccharides[J].Natural Product Research and Development, 2009, 21(1): 155-158.
[14] 张威, 朱劲华, 王敏, 等.极大螺旋藻胞内多糖的分离纯化及其结构与功能的初步分析[J].天然产物研究与开发, 2004,16(6): 548-551.DOI: 10.16333/j.1001-6880.2004.06.017.
ZHANG W, ZHU J H, WANG M, et al.Isolation and purification of intracellular polysaccharides from Spirulina maxima and preliminary analysis of its structure and function[J].Natural Product Research and Development, 2004, 16(6): 548-551.
[15] PENG F, REN J L, XU F, et al.Fractionation of alkali-solubilized hemicelluloses from delignified Populus gansuensis: structure and properties[J].Journal of Agricultural and Food Chemistry, 2010, 58(9): 5743-5750.DOI: 10.1021/jf1003368.
[16] GUAN Y, ZHANG B, QI X M, et al.Fractionation of bamboo hemicelluloses by graded saturated ammonium sulphate[J].Carbohydrate Polymers, 2015, 129: 201-207.DOI: 10.1016/j.carbpol.2015.04.042.
[17] 王慧.4种地衣多糖的分离纯化及生物活性研究[D].西安: 西北大学, 2014.
WANG H.Isolation, purification and biological activity of four Lichen polysaccharides[D].Xi'an: Northwest University, 2014.
[18] 张华峰, 李璐, 张翔, 等.5种小檗科植物多糖含量及抗氧化活性分析[J].精细化工, 2014, 31(2): 174-178.DOI: 10.13550/j.jxhg.2014.02.001.
ZHANG H F, LI L, ZHANG X, et al.Analysis of polysaccharide content and antioxidant activity of five species of polygonaceae[J].Fine Chemicals, 2014, 31(2): 174-178.
[19] 余莹, 粟武, 魏东芝.原花青素体外清除自由基活性的研究[J].华东理工大学学报, 2002, 28(3): 318-320.DOI: 10.3969/j.issn.1006-3080.2002.03.024.
YU Y, SU W, WEI D Z.Study on free radical scavenging activity of Proanthocyanidins in vitro[J].Journal of East China University of Science and Technology, 2002, 28(3): 318-320.
[20] 韦献雅, 殷丽琴, 钟成, 等.DPPH法评价抗氧化活性研究进展[J].食品科学, 2014, 35(9): 317-322.DOI: 10.7506/spkx1002-6630-201409062.
WEI X Y, YIN L Q, ZHONG C, et al.Advances in evaluation of antioxidant activity by DPPH method[J].Food Science, 2014, 35(9): 317-322.
[21] 宋丽青, 刘必谦, 王亚军, 等.氨基葡萄糖及其自然氧化产物抗氧化功能研究[J].生物学杂志, 2015, 32(3): 25-29.DOI: 10.3969/j.issn.2095-1736.2015.03.025.
SONG L Q, LIU B Q, WANG Y J, et al.Antioxidant function of glucosamine and its natural oxidation products[J].Biology Journal, 2015, 32(3): 25-29.
[22] CHEN Y, XIE M Y, NIE S P, et al.Purification, composition analysis and antioxidant activity of a polysaccharide from the fruiting bodies of Ganoderma atrum[J].Food Chemistry, 2008, 107(1): 231-241.DOI: org/10.1016/j.foodchem.2007.08.021.
[23] YE M, QIU T, PENG T W, et al.Purification, characterization and hypoglycemic activity of extracellular polysaccharides from Lachnum calyculiforme[J].Food Chemistry, 2008, 107(1): 231-241.DOI: org/10.1016/j.foodchem.2007.08.021.
[23] YE M, QIU T, PENG T W, et al.Purification, characterization and hypoglycemic activity of extracellular polysaccharides from .Carbohydrate Polymers, 2011, 86(1): 285-290.DOI: 10.1016/j.carbpol.2011.04.051.
[24] 冯书珍, 卢宇凤, 刘南英, 等.海藻多糖的单糖组成对体外抗氧化活性的影响[J].天然产物研究与开发, 2019, 169(31): 116-121.DOI: 10.16333/j.1001-6880.2019.1.018.
FENG S Z, LU Y F, LIU N Y.et al.Effect of monosaccharide composition on antioxidant activity of algal polysaccharides[J].Natural Product Research and Development, 2019, 169(31): 116-121.
[25] 倪力军, 王媛媛, 何婉瑛, 等.8种多糖的单糖组成、活性及其相关性分析[J].天津大学学报, 2014, 47(4): 326-330.DOI: 10.11784/tdxbz201207047.
NI L J, WANG Y Y, HE W Y.et al.Monosaccharide composition, activity and their correlation analysis in eight polysaccharides[J].Natural Product Research and Development, 2014, 47(4): 326-330.

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备注/Memo

备注/Memo:
收稿日期:2018-11-26 修回日期:2019-01-17
基金项目:国家自然科学基金(31470417); 北京林业大学大学生创新训练项目(S201710022048); 伽蓝(集团)股份有限公司合作项目(YF-ZX-201803-0320)。
作者简介:汪小君,女,研究方向为天然多糖的利用研究。通信作者:彭锋,男,教授。E-mail: fengpeng@bjfu.edu.cn
更新日期/Last Update: 2019-07-10