| Semen Euryales is the dry mature seed kernel of Euryale ferox Salisb.It was firstly occurred in "Shen Nong ’s Herbal Classic",and belonged to the top grade.The main producing areas of it are Jiangsu,Shandong,Hunan,Hubei,Anhui province.There are two variants of Semen Euryales classified with thorn or not.Because of the Yishen Gujing,spleen diarrhea,Qushi zhidai effect,Semen Euryales is commonly used in clinic.As a basis of National 12th five-year science and technology support plan,the project is"quality evaluation improvement and multipurpose use of Semen Euryales".With the guideline of traditional Chinese medicine theory,the study focus on quality evaluation improvement and non-traditional medicinal parts property,in the hope of multipurpose use of the staple aquatic medicinal materials——Semen Euryales.The main research includes:1 Resources,classification,nutrition,effective substance foundation,pharmacology,clinical efficacy,quality assessment method,high-yield cultivation,palynology,development and resource protection were cleared systematically by the literature research.Therefore,a new research methods for aquatic medicinal materials could be established and the mechanism for influence of water environmental variation on aquatic plants quality formation could be made out.2 The early resource findings showed that the main domestic cultivation of Semen Euryales were among Jiangsu,Shandong,Guangdong and Hubei province.Jiangsu took the good amourt of plantation part,especially by Taihu Lake Basin.There were four famous cultivation spots:Suzhou Songzeyuan aquatic vegetable base,Suzhou Loufeng Town Qunli Village,Suzhou vegetable research institute and east Lake Taihu.However,yield and quality of Semen Euryales were destroyed by the fast urbanization and water environmental deterioration.The price gone up obviously,as the fresh product could get 160 RMB/kg,and the dry one was 270 RMB/kg.The solution for this problem was to expand the cultivation area,increase productivity and improve quality.3 According to the systematical pre-trial of chemical composition,there were amino acids,proteins,sugars,phenols,organic acids and any other major categories compositions in Semen Euryales.The paper took the water-soluble compositions in depth.Semen Euryales collected from fifteen regions was analyzed for their amino acid composition and nutritional evaluation using automatic amino acid analyzer.The results showed the total amino acid content averaged in Semen Euryales was 103.33mg/g while the free amino acid was 0.98mg/g.Among human essential amino acids,the first limited amino acid was Lys,and the SRC was 66.98%.Glu,Leu were the principal components.Regions varieties showed significant differences.As the average value was 0.4577 mg/g,water-soluble protein content were low among regions.Regions varieties also showed significant differences,from high to low principle,there were Yangtze River valley,south ones,north ones,representively.Using water extraction and alcohol precipitation method to extract the Semen Euryales polysaccharide,sevage method to deproteinization,improved phenol sulfuric acid method to determine polysaccharides content,coomassie brilliant blue method to determine protein content.The result showed that purity degree of crude polysaccharides was 73.25%,while refined one was 76.22%,the protein content of crude polysaccharides was 6.27%,while refined one was 1.39%.EPJ-A1、EPJ-B1、EPJ-B2、EPJ-B3、EPJ-B4、EPJ-C1、EPJ-D1、EPJ-D2、EPJ-D3 were the fractions separated from DEAE-52,and EPJ-A1-1 和 EPJ-A1-2 were the fractions separated from Sephadex G-100.All fractions were water-soluble and in white or yellow coulor,which had no other composition except glucide refer to the physical and chemical identification.Purity test indicated that fractions separated from DEAE-52 were all impure,while EPJ-A1-1 和 EPJ-A1-2 separated from Sephadex G-100 were on equal molecular weight.90%of Semen Euryales polysaccharide were below 15 kDa molecular weight.It was made up of glucose and rhamnose,and mostly the former one,which could surmise that maybe glucoside keys constitute the complex polysaccharide.It had no cytotoxicity tested by MTT assay.So it could be raw material for food,medicines,health products research.The tannin rich in seed capsule could be economic industrial material.The optimum ultrasonic extraction technic tested by response surface methodology was:acetone concentration of 57%,Liquid ratio of 57mL/g and ultrasonic time of 23min.Collected water part and 10%ethanol part as purified tannin extract after D101 macroporous resin.Obey to analytical test methods of "Forestry industry standards of the People’s Republic of China" LY/T 1083-2008、LY/T 1082-2008,seed capsule was gone with other requirements.It needed to decolor and refine to extend the use range.4 From the whole composition point of view,it was significant to establish fingerprint of Semen Euryales.From the HPLC fingerprint,seventeen common peaks were identified in chromatograms with reference to naringenin peak from the 14 batches of the samples,which found three determined peaks and classified into five categories.Peak 1、3、4、5、13 were with strong absorption.Relative retention time was suitable,while relative peak area was varied greatly.The similarity was 0.654～0.998 compared to the mutual mode.and 0.654～0.998 compared to the control medicinal material.It was propositional to establish HPLC fingerprint beyond the similarity of 0.9,since the quality variation among origins.The naringenin content was only 1.9539±0.0143μg/g.So identification of other characteristic peaks needs more attention.On the basis of characteristic of IR fingerprint,two indexes,common peak ratio and variant peak ratio,were applied and calculated respectively,which set up a new method called dual-index sequence analysis in distinguishing samples from different regions.It showed sample S13 and S12 S14,sample S2 and S3,which from the closer regions were the most similar samples with higher common peak ratio(≥82.4%)and lower variant peak ratio(≤21.4%).Sample S9 and S1 S15,sample S4 and S6 from different regions but of same specie,were average with similar common peak ratio(≤52.4%)and variant peak ratio(≥33.3%).However,sample S16 and S8 S6,sample S17and S4 S8 from the farther regions,were of significant disparity with lower common peak ratio(≤47.6%)and higher variant peak ratio(≥50.0%).In addition,sample S10 was in different region and at closer collecting time from other samples,which resulted in much lower common peak ratio(≤35.7%)and much higher variant peak ratio(≥1 00.0%).NIR waveform and absorbance among origins were similar,as Semen Euryales had steady quality and little variation trend.1196-2332nm showed obvious characteristics which called fingerprint region could be the model of NIR spectral prediction.2331.017nm、2313.353nm、2101.412nm、1928.765nm、1761.000nm、1549.000nm、1468.118nm、1196.000nm were the mutual peaks,and 1549.000nm、1468.118nm、1761.000nm were the principal constituent peaks.5 Previous researches have filtered polysaccharide as an effective part and seed capsule polyphenol as a numerous part,the two parts could be the targets for geriatrics therapy.Anti-fatigue and hypoxia tolerance test indicated that compared to VC,VE group,poly saccharose group and testa polyphenols group of high,medium,low dose could extend mice swimming time and hypoxia tolerance time,enhance anti-stress ability evidently,and tell dose-effect relationship.In addition,activity,appetite,mental state,average daily weight increase,organ coefficient of medication administration teams were better than control group and positive groups.They could increase hepatic glycogen,muscle glycogen,serum LDH content,tissue and serum NO content,decrease serum BUN content.All above confirmed that polysaccharose and testa polyphenols did have the function of nutritions and physiological and biochemical regulation.Polysaccharose and testa polyphenols could do something good to D-galactose induced mouse aging model,which could raise tissue and serum SOD、CAT、GSH-Px、Hyp activity,reduce MAO activity,interdict Lipid peroxidation process,decrease MDA production,effectively eliminate ROS,and ease the decline of immune organs coefficient.As natural antioxidants,polysaccharose and testa polyphenols were conveniently extracted and widely exploit.In vitro antioxidant tests confirmed that polysaccharose and testa polyphenols could effectively erase·OH,O2-·,impact on Nitrosamines synthesis and Nitroso elimination,be sensitive to DPPH·and helpful to auto-oxidation of fat food,also show stronger educing power than VC and TBHQ,which may guide the therapy for diseases caused by oxygen free radicals.6 Investigated the accumulation change of active ingredients caused by environmental factors and filtered the most important one,compared the quality of each origin by grey relational model,which could determine the quality of herbal materials externally and internally and found the integrated system.Temperatures,sunshine hours and average wind speed were the influence environmental factors on naringenin,the total amino acids,water-soluble protein respectively.Semen Euryales from Zhaoqing Guangdong was the best one for its high relative relation of 0.657 measured by gray relational grade. |
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