党参残渣酸性多糖的制备、结构特征及抗氧化活性研究
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摘要
为了提高党参的利用率,缓解工业废弃物党参残渣带来的生态压力,作者从党参残渣中通过DEAE-32阴离子交换柱分离和Sepharose CL-4B凝胶柱纯化手段得到了一种具有生物活性的大分子多糖--党参残渣多糖CERP2(Codonopsis pilosula extracted residue polysaccharide 2)。通过PMP衍生化法测定其单糖组成,高效凝胶色谱、红外光谱和核磁共振波谱测定其相对分子质量和糖苷键连接方式,采用体外羟基阴离子、超氧阴离子和过氧化氢清除实验,铁离子螯合能力实验,还原力测定等实验检测其抗氧化活性。结果表明党参残渣多糖CERP2是由甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖以1.00∶4.88∶32.40∶2.35∶13.86∶19.25的比例组成的酸性多糖,其相对分子质量为1.81×106,主要由1,4-β-D-Galp、1,4-α-D-Galp A-6-OMe、1,5-α-L-Araf、1,3-α-DManp、1,6-β-D-Glcp和1,2-α-L-Rhap等糖苷键组成,并具有一定的抗氧化能力。该工作为探索党参残渣的再利用提供了新的思路。
Codonopsis pilosula [( Franch.) Nannf. ],a member of radix codonopsis,is the root of codonopsis pilosula and a perennial herb,which has immunoregulation and the spleen and lung protection effect. Because of its various pharmacological activities,it has been made into a variety of products which are mainly made from the supernatant of ethanol-precipitated Codonopsis pilosula extracted. In order to improve the utilization of the resource of Codonopsis pilosula and alleviate the ecological pressure from industrial waste of the residue of Codonopsis pilosula,a high quality polysaccharide coded as CERP2 which had specific activity was extracted and purified from the residue of Codonopsis pilosula by ethanol precipitation,DEAE-32 column chromatography,and Sepharose CL-4 B gel chromatography. The monosaccharide composition,relative molecular weight,and glycosidic bond of CERP2 were determined by PMP derivatization,HPGPC,FT-IR,and NMR,respectively. The scavenging effect on hydroxyl radical,superoxide anion and on hydrogen peroxide as well as the chelating effect with Fe3 +and the reducing power detection were used to evaluate the antioxidant activity of CERP2. The results showed that CERP2 was an acidic heteropolysaccharide composed of mannose,rhamnose,galacturonic acid,glucose,galactose,and arabinose in the ratio of 1. 00 ∶ 4. 88 ∶ 32. 40 ∶ 2. 35 ∶ 13. 86 ∶ 19. 25,which indicated that Galacturonic acid was the majority monosaccharide in CERP2. HPGPC showed that CERP2 was homogeneous with a relative molecular weight of 1. 81 × 106. The result of infrared spectra confirmed that both α-configuration and β-configuration were existing in CERP2.The results of NMR indicated that 1,4-linked β-D-Galp,1,4-linked α-D-Galp A-6-OMe,1,5-linked α-L-Araf,1,3-linkedα-D-Manp,1,6-linked β-D-Glcp,and 1,2-linked α-L-Rhap were the main linkages in CERP2. The antioxidant experiments in vitro indicated that CERP2 had potential antioxidant ability especially in the scavenging effect on hydroxyl radical,hydrogen peroxide and the chelation with Fe3 +. This work provides new thoughts and data support for exploring the residue of Codonopsis pilosula.
Codonopsis pilosula [( Franch.) Nannf. ],a member of radix codonopsis,is the root of codonopsis pilosula and a perennial herb,which has immunoregulation and the spleen and lung protection effect. Because of its various pharmacological activities,it has been made into a variety of products which are mainly made from the supernatant of ethanol-precipitated Codonopsis pilosula extracted. In order to improve the utilization of the resource of Codonopsis pilosula and alleviate the ecological pressure from industrial waste of the residue of Codonopsis pilosula,a high quality polysaccharide coded as CERP2 which had specific activity was extracted and purified from the residue of Codonopsis pilosula by ethanol precipitation,DEAE-32 column chromatography,and Sepharose CL-4 B gel chromatography. The monosaccharide composition,relative molecular weight,and glycosidic bond of CERP2 were determined by PMP derivatization,HPGPC,FT-IR,and NMR,respectively. The scavenging effect on hydroxyl radical,superoxide anion and on hydrogen peroxide as well as the chelating effect with Fe3 +and the reducing power detection were used to evaluate the antioxidant activity of CERP2. The results showed that CERP2 was an acidic heteropolysaccharide composed of mannose,rhamnose,galacturonic acid,glucose,galactose,and arabinose in the ratio of 1. 00 ∶ 4. 88 ∶ 32. 40 ∶ 2. 35 ∶ 13. 86 ∶ 19. 25,which indicated that Galacturonic acid was the majority monosaccharide in CERP2. HPGPC showed that CERP2 was homogeneous with a relative molecular weight of 1. 81 × 106. The result of infrared spectra confirmed that both α-configuration and β-configuration were existing in CERP2.The results of NMR indicated that 1,4-linked β-D-Galp,1,4-linked α-D-Galp A-6-OMe,1,5-linked α-L-Araf,1,3-linkedα-D-Manp,1,6-linked β-D-Glcp,and 1,2-linked α-L-Rhap were the main linkages in CERP2. The antioxidant experiments in vitro indicated that CERP2 had potential antioxidant ability especially in the scavenging effect on hydroxyl radical,hydrogen peroxide and the chelation with Fe3 +. This work provides new thoughts and data support for exploring the residue of Codonopsis pilosula.
引文
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[13]Zhu W,Xue X,Zhang Z.Structural,physicochemical,antioxidant and antitumor property of an acidic polysaccharide from Polygonum multiflorum[J].Int J Biol Macromol,2017,96:494-500.
[14]Zhang BW,Xu JL,Zhang H,et al.Structure elucidation of a polysaccharide from umbilicaria esculenta and its immunostimulatory activity[J].PLo S One,2016,11(12):e0168472.
[15]郗红娟,靳岚.核磁共振波谱法在药物分析中的应用[J].药物生物技术,2013,20(5):453-457.
[16]朱恩圆,贺庆,王峥涛,等.党参化学成分研究[J].中国药科大学学报,2001,32(2):2.
[17]周茜,吴梦颖,王唯霖,等.多糖型扇贝沙司对小鼠免疫功能和抗氧化活性的影响[J].中国食品学报,2018,2:30-36.
[2]Tang Q,Yang S,Tong J,et al.Hemostasis and uterine contraction promoting effect of the extract from drugs in the Zi-Yin-Tiao-Jing granule,a traditional Chinese compound preparation[J].J Ethnopharmac,2018,211:278-284.
[3]Qi FH,Zhao L,Zhou AY,et al.The advantages of using traditional Chinese medicine as an adjunctive therapy in the whole course of cancer treatment instead of only terminal stage of cancer[J].Biosci Trends,2015,9(1):16-34.
[4]Liu W,Liu YM,Zhu R,et al.Structure characterization,chemical and enzymatic degradation,and chain conformation of an acidic polysaccharide from Lycium barbarum L[J].Carbohyd Polym,2016,147:114-124.
[5]Liu W,Lu WS,Yin C,et al.Preliminary structural characterization and hypoglycemic effects of an acidic polysaccharide SERP1from the residue of sarcandra glabra[J].Carbohyd Polym,2017,176:140-151.
[6]Kang MC,Kim SY,Min YT,et al.In vitro and in vivo antioxidant activities of polysaccharide purified from aloe vera(Aloe barbadensis)gel[J].Carbohyd Polym,2014,99:365-371.
[7]Kan YJ,Chen TQ,Wu YB,et al.Antioxidant activity of polysaccharide extracted from Ganoderma lucidum using response surface methodology[J].Int J Biol Macromol,2015,72:151-157.
[8]Gao J,Zhang T,Jin ZY,et al.Structural characterisation,physicochemical properties and antioxidant activity of polysaccharide from Lilium lancifolium Thunb[J].Food Chem,2015,169:430-438.
[9]Chen L,Zhang YP,Jin LX,et al.Preparation,characterization and antioxidant activity of polysaccharide from Fallopia multiflora(Thunb.)Harald[J].Int J Biol Macromol,2018,108:259-262.
[10]Sun YX,Liu JC.Structural characterization of a water-soluble polysaccharide from the roots of Codonopsis pilosula and its immunity activity[J].Int J Biol Macromol,2008,43(3):279-282.
[11]Zhang YJ,Zhang LX,Yang JF,et al.Structure analysis of watersoluble polysaccharide CPPS3 isolated from Codonopsis pilosula[J].Fitoterapia,2010,81(3):157-161.
[12]Zou YF,Chen XF,Malterud KE,et al.Structural features and complement fixing activity of polysaccharides from Codonopsis pilosula Nannf.var.modesta L.T.Shen roots[J].Carbohyd Polym,2014,113:420-429.
[13]Zhu W,Xue X,Zhang Z.Structural,physicochemical,antioxidant and antitumor property of an acidic polysaccharide from Polygonum multiflorum[J].Int J Biol Macromol,2017,96:494-500.
[14]Zhang BW,Xu JL,Zhang H,et al.Structure elucidation of a polysaccharide from umbilicaria esculenta and its immunostimulatory activity[J].PLo S One,2016,11(12):e0168472.
[15]郗红娟,靳岚.核磁共振波谱法在药物分析中的应用[J].药物生物技术,2013,20(5):453-457.
[16]朱恩圆,贺庆,王峥涛,等.党参化学成分研究[J].中国药科大学学报,2001,32(2):2.
[17]周茜,吴梦颖,王唯霖,等.多糖型扇贝沙司对小鼠免疫功能和抗氧化活性的影响[J].中国食品学报,2018,2:30-36.