Study On Preparation Of Arctiin And Arctigenin

Fructus Arctii is the dried ripe seeds of Compositae-- Arctium lappl L, which is one kind of traditional chinese medicine. Many studies, such as pharmacokinetics research and so on, indicated that arctii and arctigenin is the major active chemical constituent of Fructus Arctii. But the content of Arctii and arctigenin in Fructus Arctii are very low, and the preparetion of them was difficult. How to improve effectively arctii and arctigenin content is the seeking aim. In this study, Arctii and arctigenin were prepared by microorganism fermentation, ethanol reflux extraction, and silica-column chromatography. The research mainly includes:Firstly, study on the method of extraction of Fructus Arctii and purification of arctii and arctigenin. The study of extraction of Fructus Arctii using ethanol refluxing method showed that the optimum extraction conditions is using 80% ethanol,1 hour of reflux,8-fold solvent and 3 times extraction. Arctii andarctigenin were isolated and purified by silica-column chromatography, and eluted by CH2Cl2: CH3OH(20:1), CH2Cl2:C4H8O2:CH3OOH (4:4:1), CH2Cl2:C4H8O2 (5:4) successively. The purity of arctii and arctigenin was attained 95.5% and 94% respectively. Secondly, a high-yield strain of producingβ-glucosidase is isolated from musty Fructus Arctii using microbial screening techniques. The optimal conditions of producingβ-glucosidase were obtained in this study. That was that the rotation speed was 200 r/min, culture temperature was 30℃, and culture time was 7 d. The enzyme activity reached 15 U/ml.Thirdly, the preparation of arctigenin by hydrolysis of arctii byβ-glucosidase which was screened was studied. The optimum conditions of hydrolytic arctii by P-glucosidase were:hydrolytic temperature 50℃, hydrolytic time 36 h, with the addition of 200 mmol/L Mg2+ at 6.0 pH, rotation speed 200 r/min. The yield of arctigenin was reached 94%.At last, the fermentaion preparation of arctigenin was studied by fermentation media which using Fructus Arctii as only nutrient. The optimizing of fermentation condition was attained by the direction of L9 (34) orthogonal experimental design:solid to liquid ratio was 1:10(g/ml), initial pH of medium was 6, rotation speed was 220 r/min, culture temperature was 30℃, culture time 7 d. The yield of arctigenin was reached 78.6%.