Study On Seperation And Purification Active Ingredient And Its Functionality Of Kalimeris Indica (L.) Sch-Bip

Kalimeris indica (L.) Sch-Bip,asteraceae,kalimeris,perennial ratoon herb,has high nutritional and medicinal value.Kalimeris is divided into a number of varieties,and each of them has different leave shape and color of immature stem.Ten kinds of Kalimeris (M1-M10) were selected as study subjects in the present study.Their main nutrient contents were analysed preliminarily.We isolated and purified M4,, then study its active components’ inhibition effect of non-enzymatic protein glycosylation.The systematic research of M1-M10 in this study can provide theoretical foundation for Kalimeris indica processing and promote the economic benefits of Kalimeris indica planting.β-carotene were measured using GB methods; polyphenol contents were measured by Folin-Ciocalteus method; flavonoid contents were measured by sodium nitrite-aluminum nitrate-sodium hydroxide chromogenic method; saponins substances were measured by vanillin-sulfuric acid colorimetric method.And the activity about anti non-enzymatic glycosylation were analysed using fluorimetry method.The result shows that Kalimeris is rich in nutrition ingredients.β-carotene content of M2 and M5 are the highest;Polyphenol and flavonoid content of M3 and M10 are the highest;saponins content of M4 and M6 are the highest.By analysing the anti non-enzymatic glycosylation effect of these ten kinds of Kalimeris’crude extract,we come to the conclusion that all of them have good inhibition effect and the inhibition rate of GO and MGO causing non-enzymatic glycosylation are 52.2% and 61.2%.The inhibition of non-enzymatic glycosylation caused by GO has positive correlation with polyphenol content and flavonoid content, and inhibition of non-enzymatic glycosylation caused by MGO has positive correlation with saponins content. Based on all the index,M4 was selected for further study.M4 was separated and purified using AB-8 macroporous resin and Sephadex LH-20.We identified that Kalimeris contain 3,5-dicaffeoylquinic acid and 3,4,5-tris-caffeoylquinic acid by HPLC-MS, and obtained a compound eventurally by repeated purification over Sephadex LH-20 column.The compound was identified as 3,5-dicaffeoylquinic by HPLC-MS and NMR.Anti non-enzymatic glycosylation of components purified by macroporous resin were studied by fluorimetry method.They were 20% ethanol fractions eluted(F1);40% ethanol fractions eluted(F2);60% ethanol fractions eluted(F3); 80% ethanol fractions eluted(F4) and 3,5-caffeoylquinic acid using fluorimetry.The result shows that F2 is the most effective and its nhibition rate of non-enzymatic glycosylation caused by GO and MGO are 78% and 76%.3,5-dicaffeoylquinic purified from F2 is also the most effective one and its inhibition of non-enzymatic glycosylation caused by GO and MGO are 92% and 95% respectively.