Purification And Antioxidant Activities Of Polysaccharides From Matricaria Chamomilla And Chrysanthemum Morifolium

Matricaria chamomilla and Chrysanthemum morifolium were all belong to annual or perennial herbaceous plant of the compositae family.Matricaria chamomillais widely cultivated and consumed worldwide,Chrysanthemum morifolium mainly cultivated in the Zhejiang province.In this study,the extraction,purification,characterization,and antioxidant activities of polysaccharides from Matricaria chamomilla and Chrysanthemum morifolium were systematically studied.The conclusions were as follows:1.Extraction process of polysaccharidesIn order to determine the optium extraction method of polysaccarides from Matricaria chamomilla and Chrysanthemum morifolium,the effects of different extraction method were researched,using water as extraction solvent.The results indicated that the polysaccharide yields from Matricaria chamomilla extracted with ultrasonic extraction(UE),microwave-assisted extraction(ME)and hot reflux extraction(HRE)were higher than other the methods,the polysaccharide yields from Chrysanthemum morifolium extracted with ultrasonic extraction and microwave-assisted were higher than the other methods.Single factor experiment and Orthogonal Experiment Assistant were employed to optimize the corresponding extraction process of polysaccharides.And the results were as follows:(1)The optimal UE conditions of polysaccharides from Matricaria chamomilla were determined as followings: ultrasonic temperature 50?,ultrasonic time 30 min,ultrasonic power 1050 W and the liquid-solid ratio 30 mL/g.Under these conditions,the extraction yield was 5.53%.(2)The optimal ME conditions of polysaccharides from Matricaria chamomilla were determined as followings: extraction temperature 50?,extraction time 30 min,extraction power 500 W,water to material ratio 40 mL/g.Under these conditions,theextraction yield was 4.70%.(3)The optimal HRE conditions of polysaccharides from Matricaria chamomilla were determined as followings: hot reflux temperature 60 ?,hot reflux 2.0 h,extraction times 1,water to material ratio 20 mL/g,and the extraction yield was 3.58%.(4)The optimal UE conditions of polysaccharides from Chrysanthemum morifolium were determined as followings: ultrasonic temperature 60 ?,ultrasonic time 30 min,ultrasonic power 450 W,the liquid-solid ratio 40 mL/g.and the extraction yield was 12.55%.(5)The optimal ME conditions of polysaccharides from Chrysanthemum morifolium were determined as followings: extraction temperature 50?,extraction time 30 min,extraction power 300 W,water to material ratio 30 mL/g,and the extraction yield was 9.75%.2.Purification and characterization of polysaccharidesThe polysaccharides(MC and CM)were purified by DEAE-Sepharose CL-6B and Sephadex G-100 chromatography.The polysaccharide extracted by ultrasound extraction of from Matricaria chamomilla(MCU)was isolated and purified to obtain two homogeneous components,named as MCU-1-1 and MCU-2-1.The polysaccharides extracted by microwave extraction from Matricaria chamomilla(MCM)was isolated and purified to obtain two homogeneous components,named as MCM-1-1 and MCM-2-1.The polysaccharides extracted by reflux extraction from Matricaria chamomilla(MCH)was isolated and purified to obtain two homogeneous components,named as MCH-1-1 and MCH-2-1.At the same time,the polysaccharide of Chrysanthemum morifolium by ultrasound extraction(CMUP)was isolated and purified to obtain two homogeneous components,named as CMUP-1-1 and CMUP-2-1.Under the condition of microwave extraction,the Chrysanthemum morifolium polysaccharide(CMMP)was isolated and purified to obtain twohomogeneous components,which were named as: CMMP-1-1 and CMMP-2-1,respectively.The total polysaccharide contents of ten polysaccharides were MCU-1-1(88.16%),MCU-2-1(82.14%),MCM-1-1(87.52%),MCM-2-1(85.24%),MCH-1-1(82.23%),MCH-2-1(84.09%),CMUP-1-1(81.13%),CMUP-2-1(83.17%),CMMP-1-1(86.02%),CMMP-2-1(81.74%),respectively.The protein contentsof ten polysaccharides were MCU-1-1(0.75%),MCU-2-1(0.66%),MCM-1-1(0.59%),MCM-2-1(1.03%),MCH-1-1(0.58%),MCH-2-1(0.72%),CMUP-1-1(0.86%),CMUP-2-1(1.05%),CMMP-1-1(0.99%),CMMP-2-1(1.15%),respectively.They all contain higher amounts of uronic acid: MCU-1-1(5.41%),MCU-2-1(8.25%),MCM-1-1(7.42%),MCM-2-1(8.94%),MCH-1-1(4.11%),MCH-2-1(5.38%),CMUP-1-1(5.80%),CMUP-2-1(7.30%),CMMP-1-1(4.09%),CMMP-2-1(7.13%),respectively.The protein contents were relatively low.Ten polysaccharides fractions showed very high solubility,and there were no significant difference in solubility between them.The results showed that different extraction were beneficial to increase the water solubility of polysaccharides.Analyses of Congo red test indicated that there was triple-helix structure existed in the MCM-2-1,MCU-2-1,CMUP-2-1 and CMMP-2-1.And the other polysaccharides fractions were no triple-helix structure existed.Thermal characteristic results indicate that ten polysaccharides fractions displayed favorable thermal stability.The molecular weight of ten polysaccharides were determined by HPGPC,and the results were as follows: MCU-1-1(141253 Da),MCU-2-1(36307 Da),MCM-1-1(72443 Da),MCM-2-1(7413 Da),MCH-1-1(30902 Da),MCH-2-1(7244 Da),CMUP-1-1(53703 Da),CMUP-2-1(7585 Da),CMMP-1-1(52480 Da),CMMP-2-1(16595 Da),respectively.FT-IR showed that these samples all had characteristic absorption peak ofpolysaccharides.These polysaccharides were composed of rhamnose(Rha),glucuronic acid(GlcA),galactose(Gal),glucose(Glu),galacturonic acid(GalA),xylose(Xyl)and arabinose(Ara)with different molar ratio.MCU-1-1 was composed of Rha,Gal,Glc,GalA,Xyl and Ara in a mole ratio of 6.93,29.69,15.61,12.76,14.12,20.88.MCU-2-1 was composed of Gal,GalA,Xyl and Ara in a mole ratio of78.67,7.65,8.29,5.39.MCM-1-1 was composed of Rha,Gal,Glc,GalA and Ara in a mole ratio of 2.05,20.99,61.21,4.22,11.53.MCM-2-1 was composed of Gal,GalA and Ara in a mole ratio of 85.38,7.03,7.59.MCH-1-1 was composed of Rha,GlcA,Gal,Glc,GalA,Xyl and Ara in a mole ratio of 6.29,2.74,4.31,5.58,14.89,11.73,54.23.MCH-2-1 was composed of GlcA,Gal,Glc,GalA,Xyl and Ara in a mole ratio of 5.53,52.88,4.26,12.24,10.21,6.32.CMUP-1-1 was composed of Rha,Gal,Glc,GalA and Ara in a mole ratio of 1.40,50.96,11.20,10.30,16.13.CMUP-2-1 was composed of Gal,Glc,GalA and Ara a mole ratio of 67.2,5.10,10.06,2.43.CMMP-1-1 was composed of Rha,Gal,Glc,GalA and Ara in a mole ratio of 2.04,68.20,5.26,10.74,6.48.CMMP-2-1 was composed of Gal,Glc,GalA and Xylin a mole ratio of 52.60,3.98,5.82 and 4.06.3.The antioxidant activity of polysaccharidesThe antioxidant activities of ten polysaccharides were determined by scavenging?DPPH and ? OH test.And the results showed that ten polysaccharides have strong antioxidant activities.And their antioxidant activities were correlated with the concentration of polysaccharide.The results showed that the extraction method had no significant effect on the sugar bond type and conformation of polysaccharides,but it might affect the molecular weight and the content of glyoxylic acid,which indirectly affected the biological activity of polysaccharides.