The Extraction And Separation Of The Isoflavone In The Root Of Amorpha Fruticosa L. And The Study On α-Glucosidase Inhibitory Activity

Amorpha fruticosa L. belongs to Leguminosae and Amorpha. The flowers of Amorpha fruticosa L.can clear heat, cooling blood, bleeding; leaves are slightly bitter, cold and has the effect of dampness and swelling, and Amorpha fruticosa L. can heal the carbuncles, eczema, burns. This paper studied the root of Amorpha fruticosa L. Systematicly including extraction process, purification process, the separation of chemical constituentsof the total isoflavones, α-glucosidase inhibiting activity and other fields to provide references for further development and utilization.By reviewing the relevant literature at home and abroad, this paper summarized chemical constituents and pharmacological effects of Amorpha fruticosa L. to provide references for further study.As an indicator to calycosin7-O-β-D-glucopyrano-side, isoflavones of the extract were determined by UV spectrophotometry. And by single factor experiment, the main factors of the extraction rate of total isoflavones were defined, and the optimum conditions were optimized on total isoflavones of the root of Amorpha fruticosa L. using Box-Behnken response surface method. The optimum conditions: ethanol concentration was 70%, solid-liquid ratio was 1:13, extraction time was 3 h, extracted for 3 times, and extraction rate of total isoflavones was 5.59%. By examining the different types of macroporous resin, D101 macroporous resin was detemined to purificate the total isoflavones, and the content of total isoflavones purificated were more than 70%.This reserch established HPLC characteristic spectrum of total isoflavones, and isolated and purified the total isoflavones, furthure the experiment got 6 compound which were calycosin7-O-β-D-glucopyrano-side, 7,3’-dihydroxy-4’-methoxyisoflavone, Dalbinol, amorphigenin, 7-dihydroxy-4’-methoxyisoflavone, 4’,7- dihydroxy-7-O- glucopyranoside.In the end, this paper screened the inhibition of α-glucosidase in vitro in the ethanol extract of the root of Amorpha fruticosa L.、purificated total isoflavones and 6 compounds using PNPG for the substrate, α-glucosidase from the small intestine of rats for reaction enzyme. By calculated IC50 using logarithmic regression curve, the resules showed that the purificated total isoflavone exceeded the the positive control drug acarbose on α-glucosidase inhibitory activity, Dalbinol、amorphigenin were similar to acarbose.Innovation:1. The first time to study the active ingredients of the root of Amorpha fruticosa L. on α-glucosidase Systematicly;2. The first time to determine the active ingredientsof the root of Amorpha fruticosa L. on α-glucosidase;3. The yield of the purificated total isoflavone is 5.4%, the contents are more than 70%, conforming the requirements of traditional Chinese Ⅴnew drugs and laying the foundation for the development of new drugs;4. Through studying, the root of Amorpha fruticosa L. which are new medicinal resources are identified, and preparating α-glucosidase inhibitor medicine.