Purification And Preparation Of Oleuropein

Oleuropein is a non-toxic iridoid glycoside compounds derived from the leaves of olive tree with molecular formula C₂₅H₃₂O₁₃;Studies have shown that oleuropein has many effects,such as anti-inflammatory,antifungal,antiviral,antioxidant,anticancer and hypoglycemic.In this paper,the purity of olivine was used as an index,and the conditions of purification by silica gel column chromatography,dextran gel chromatography and preparative chromatography were studied in detail.The purification technology parameters were optimized by response surface methodology,and the high-purity products were obtained.The ¹H-QNMR value and antioxidant capacity of oleuropein in vitro were further studied.Firstly,the ultrasonic extraction conditions of oleuropein were investigated in detail.The optimal extraction conditions were determined as follows:45 min,45℃,200 W;The HPLC method for analysis of oleuropein was determined.The process parameters of silica gel column chromatography were studied by response surface analysis.The optimal conditions were determined as follows:column diameter of 3cm,ratio of diameter:height 1:10,sample weight of 5 g,eluent of ethyl acetate:methanol:water（19:1:0.3）;Under these conditions,the purity of oleuropein reached 85%（area normalization method）.In order to improve the purity of the product,secondary column chromatography was studied.The purity of the product was 96%（area normalization method）,which was higher than the purity reported in literature.In order to obtain higher purity of oleuropein,preparative chromatography was used.The optimal conditions were as follows:mobile phase was methanol:water（40:60）,flow rate was 15 m L/min,injection volume was 2 ml.The purity of oleuropein product could reach 99.9%（area normalization method）.The purity of oleuropein purified by secondary column chromatography was studied by 1H NMR.The experimental parameters were optimized and the purity was determined to be 93.75%.Finally,the antioxidant activity of oleuropein was studied in vitro.The results indicated that oleuropein had good antioxidant activity in vitro.