Preparation Of Polysaccharides From Chlorella Vulgaris And Studies On Bioactive Components From The Phragmites Australis

In this paper, the chemical constitutes and biological activity from Chlorellavulgaris and the aerial of Phragmites australis were studied by using variouschromatography analysis and recrystallation methods. The extraction processes ofactive constituents were optimized.After optimizing the extraction methods of polysaccharides from Chlorellavulgaris, flash extraction combined with enzymolysis method was the best for theextraction of chlorella polysaccharides for the highest extraction ratio of digestionauxiliary by flash extraction, and trypsin was chosen from six kinds of enzymes.The optimal process conditions were determined as1.5%trypsinase, pH7.5,40℃and2h for enzyme dosage, pH value, enzymolysis temperature and time. Besides,the protein was removed and fractional precipitation with ethanol, among which thepart ethanol precipitation was80%has been sepatated and purificated by DEAE-52.At last, chlorella polysaccharide was purified at a rate of3.75‰. The chlorellapolysaccharides had some inhibition of the migration of induced human umbilicalvein endothelial cells at the concentration of50and100μg/mL.13compounds were separated from petroleum ether extract, ethyl acetateextract and n-butanol extract of phragmites australis by a variety of chromatographictechniques, physical and chemical constant determination, spectroscopic dataanalysis,7compounds were identified as β-sitosterol (L-1),4-Hydroxybenzaldehyde(L-2), Tricin (5,7,4’-trihydroxy-3’,5’-dimethoxy flavone,L-3), Tricin7-O-β-glucopyranoside (L-4), Glycerol alpha-monopalmitate (L-5),n-butyl-O-β-D-fructopuranoside (L-6) and daucosterol (L-7). The compounds L-3,L-4, L-5and L-6were isolated from phragmites australis for the first time.The ACE enzyme inhibitory activity was determined on the alcohol extract,petroleum ether extract, ethyl acetate extract and n-butanol extract from phragmitesaustralis. The inhibitory activities of each extract increased with the increase of theirconcentration. Among them, the n-butanol extract had a better inhibitory activity as awhole. The inhibition rate can reach71.6%at the concentration of100mg/mL. Thepetroleum ether extract had also some inhibitory activity.To get the total flavonoids from phragmites australis, it was extracted with95%ethanol by using heat reflux extraction combining with flash extraction. The bestconditions were material to liquid was0.7:100g/mL,5minutes of flash extractionand then2hours of heat reflux extraction. Due to a large number of tricin wasseparated, which was as standard when measuring the flavonoids content. Finally,5 kinds of macroporous resins were tested to purify the flavonoid parts, amongthem AB-8and HPD100had high ratios at both adsorption and desorption for totalflavonoids. The sample concentration and its pH were studied for HPD100, when theconcentration was0.53mg/mL and pH at6, the adsorption ratio was21.1%and32.6%, respectively.It might be provide some theoretical foundation for researching Chlorellavulgaris and Phragmites australis fully.