Study On Separation Technology Of Efficient Extraction Of Active Ingredients From Jerusalem Artichoke

Object:Helianthus tuberosus L., known as the Jerusalem artichoke, is a perennial herbaceous plant which belongs to the genus of Asteraceae of family Helianthus.This thesis focused onthe high efficient extract and separation technique s tudies of active component phenolic acid from the aerial part of Jerusalem artichoke.Methods: A simple process combining ultrasonic extract and novel aqueous two phases partition was successfully developed and technological parameter was optimized.1. HPLC method was first developed to detect phenolic acid in sample. The HPLC separation was carried out on a cosmosil C18(250×4.6mm) column. andacetonitrile(A)-0.1% formic acid(B) as mobile phase with a gradient elution. 0-10 min,10~15%A;10-25 min,15~45%A;25-50 min,45~100%A. flow rate was 1 ml/min and column was kept under 30°C. Detected wavelength was set as 327 nm.2. Ultrasonic wave extract was used to extract active component from Helianthus tuberosus L. The total content of chlorogenic acid(CQA) and 1,5-dicaffeoyquinic acid(DiCQA) in extract was chose as evaluation index to optimized extract condition. The orthogonal experiment L 9(34) was arranged with four factors, which were extraction time, extraction times,ethanol content and solvent multiple, and were arranged into three levels respectively.3. Novel aqueous two phases partition was applied to enrich active component in sample. The distribution ratio of CQA and DiCQA in partition solvent was chose as evaluation index to optimized fraction partition condition. Content of ethanol, water, and ammonia sulfate was optimized and best condition was chose to enrich active component in sample.4. Chemical and antioxidant activity of sampleThe active component content in final product was determinated with HPLC method and antioxidant activity of sample was evaluated with DPPH and ABTS method。Results:1. HPLC method was successfully developed to detect phenolic acid in sample. Active components were baseline separation in 30 running time. Compared with standard sample. CQA and DiCQA was the two main component in Helianthus tuberosus L and the content of CQA and DiCQA was 0.48 and 2.19%, respectively.2. The orthogonal experiment result showed the best extract parameters were 15 time 60% ethanol extract two times,10 min per time.3. The partition system of ethanol, water, andammonia sulfate(30:49:21,W/W/W)was the best solvent to enrich active component in sample. Partition coefficient of chlorogenic acid and 1,5-dicaffeoyquinic acid was 7.43 and 13.75.4. The optimizedextract and partition method was applied to product active components from aerial part o f Helianthus tuberosus L. The product contained 13.37% CQA and 7.1% DiCQA. The recovery of them was 88.44% and 94.47%, respectively. The of Jerusalem artichoke extract and active extracts for antioxidant activity compare, which obtained by a novel aqueous two-phase extraction of the active site of antioxidant capacity significantly increased to 25.75 μg/ml 4.91 μg/ml DPPH and ABTS method.