Study On The Extraction And Purification Process Of Angiotensin-I Convert Enzyme Inhibitor In Grifola Frondosa

Grifola frondosa is a kind of delicious mushroom abundant in physiological functions, its active compositions can resist hypertension and adiposity, prevent anaemia, scurvy, arteriosclerosis, cephalitis , and counteract caducity and cancer. The anti-hypertension composition in Grifola frondosa had been proved a peptide which can inhibit angiotensin-I concert enzyme(ACE) . This thesis studied the extraction and purification of ACE inhibitor from Grifola frondosa.The procedure to analyze the activity of ACE inhibitor with HPLC was developed. The activity of ACE inhibitor was assayed by Dikma DiamonsilTM C18 column (250mm×4.6mm, 5μm) with acetonitrile-0.2% phosphoric acid (30:70) as mobile phase and detected with UV 228nm and a flow rate of 1.000mL min-1. Results showed that is a novel, prompt, concise and useful method for estimating the angiotensin-I converting enzyme inhibitory activity.The procedure to analyze the ACE inhibitor with HPLC was set up. The contents of ACE inhibitor in the plant of Grifola frondosa were assayed by Dikma DiamonsilTM C18 column (250mm×4.6mm,5μm) andgradient eluted with acetonitrile-water system(30-%70B/15min) and detected with UV 280nm and a flow rate of 1.000mL·min-1. Results showed that is a prompt and concise method for estimating the angiotensin-I converting enzyme inhibitory.The optimal extracting condition was obtained. The crude product was obtained with a content of 2.49% and a yield of 93.6% at the condition of that the raw materials was extracted two times for 6h at 30℃ with 40% deionized water and the ratio (solvent: raw materials) of 20:1.The purification process was investigated and different kinds of products (8.0%, 25.0%, 35.1%, above 97.5% in content) were obtained by the means of adsorption and desorption with SP700 resin, extraction by gel filter chromatography and HPLC chromatography.The technology conditions for adsorbing and separating ACEI with SP700 resin were as follows: the sample of Grifola frondosa extracts were centrifuged at 4℃ at 1000×g, the supernatants were adsorpted by SP700 at 50ml/h and also eluted with deionized water. ACE inhibitor was not adsorbed by SP700 at all, the yield of product was 100%, ACE inhibitor showed filemot and a content of 8.0%.The liquid after SP700 was applied to Sephadex G-50 column and eluted with Tris-HCl buffer at 12ml/h, the yield was 67.5% and the content of ACE inhibitorby was 25.0%. The active fractions obtained were then subjected to column chromatography on a Sephadex LH-20 in 15%...