Optimization Of The Extraction Technology For The Active Ingredient In Anemarrhena Asphodeloides Bunge And The Study The Active Ingredient In Hypoglycemic Activity

Diabetes mellitus (DM) has become a chronic diseases after tumor and cardiovascular disease mortality rates are the highest. Anemarrhena asphodeloides Bunge has obvious therapeutic effect on diabetes, so study the chemical constituents, optimization of the extraction technology for the active ingredient in Anemarrhena asphodeloides Bunge and the active ingredient in hypoglycemic activity has important academic value and practical significance.First of all, using column chromatography separation technology and UV, IR and NMR technology, six compounds were isolated:neomangiferin, mangiferin, timosaponin AⅡ, timosaponin A Ⅲ, timosaponin B Ⅱ and timosaponin BⅢ. A fast, high sensitivity, strong specificity and accuracy RP-HPLC-ELSD method was optimized and validated for determination of four compounds (neomangiferin, mangiferin, timosaponin B Ⅱ and timosaponin BⅢ) in Anemarrhena asphodeloides Bunge. This method is suitable for quantitative analysis and quality control of Anemarrhena asphodeloides Bunge medicinal materials.Second, using the hot refluxing extraction, the content of neomangiferin, mangiferin, timosaponin B Ⅱ and timosaponin B Ⅲ in Anemarrhena asphodeloides Bunge as indicators, using single factor method examines different solvent systems, soak time, material liquid ratio, extraction time, medicinal material particle size, pH value and extraction times on the effect of active ingredients in Anemarrhena asphodeloides Bunge extract, and obtain the optimal extraction process of active ingredient in Anemarrhena asphodeloides Bunge:medicinal material particle size(60-100mesh), a certain volume of75%ethanol (pH11.0) according to the material liquid ratio1:8was added, soak20min, hot reflux extraction of3h, extracting3times, collecting the extracted liquid, evaporating the solvent, and obtain75%ethanol extract which contains the contents of neomangiferin, mangiferin, timosaponin BⅡ and timosaponin B Ⅲ were5.82%,4.33%,38.67%and15.55%, respectively.Finally, building alloxan diabetes mice model, the different amount of75%ethanol Anemarrhena asphodeloides Bunge extract and timosaponin A Ⅲ are given respectively into stomach of normal mice and glycermia mice induced by alloxan on time each day for28days. The experimental results show that75%ethanol Anemarrhena asphodeloides Bunge extract and timosaponin AⅢ can improve the antioxidant ability of normal and alloxan diabetic mice by improving the liver SOD activity and reducing the MDA content, and also play a role of insulin sensitization, promote glucose utilization, promote the synthesis of glycogen, inhibiting glycogen dysplasia, so as to regulate sugar metabolism, reduce blood sugar level, improve symptoms of diabetes.