Enrichment Of N-feruloylserotonin And N-(p-coumaroyl)Serotonin In Safflower(Carthamus Tinctorius L.) Seed Meal

The N-feruloylserotonin (FS) and N-(p-coumaroyl)serotonin (CS) in safflower (Carthamus tinctorius L.) seed meal exhibited anti-oxidation, anti-inflammatory and anti-tumor effect, can be used in food, medicine, cosmetics and other fields. They have broad development prospects. In this paper, enrichment process of N-feruloylserotronin (FS) and N-(p-coumaroyl)serotonin (CS) in safflower (Carthamus tinctorius L.) seed meal has been researched systematically.Through single factor experiment and orthogonal experiment, the optimal technology of extracting the total phenylpropanoid amides of 5-hydroxytryptamine (PAHA) from meal was determined as follow: 60% ethanolas extraction solvent, extracted at 65℃for 90min, solid-liquid ratio of 1:20, grinding granularity of 35 mesh. The yield of PAHA was 0.784%, and the purity was 7.87%; The yield of the crude extraction was 9.96%, and the purity of FS, CS, FSG and CSG in the crude extraction were 3.82, 2.58, 0.53 and 0.94% respectively. The shrinking core model on PAHA extraction of safflower seed meal by 60% ethanol was established, based on the Fick's first law. According to the experimental data, the model was solved and validated. The extraction rate-determining step of the process was the diffusion of solute through the extracted residual layer. And The extraction time t was in the good linear relationship with . The results proved that the model could be used to analyze and predict the process of extracting PAHA from safflower seeds by ethanol.The optimal enzymolysis conditions of PAHA glucoside were detemined by using the single factor experiment and response surface analysis:The crude extract concentration of PAHA 3.3g/L, pH=5.9, enzymolysis with 3.00% (g/g crude extraction)β-D-glucosidase under 47℃f or 10.5 hr. The conversation rate of N-(p-coumaroyl)serotonin-mono-β-D- glucopyranoside (CSG) and (N-feruloylserotronin)-mono-β-D-glucopyranoside (FSG) were 94.6 and 90.0% respectively, and the FS and CS content in the conversation product were 3.42 and 4.30%.After enzymatic hydrolysising PAHA crude extraction byβ-D-glucosidase, enrichment processes of FS and CS with D818 weak alkaline macropore anion ion-exchanger resin were studied. The optimal adsorption and desorption experiments on D818 resin were conducted to establish parameters as: FS and CS concentration in sample solution 30-50 and 25-41mg/mL, respectively, pH=9.1(for adsorption), sample flow rate 1.5 mL/min; eluent 80% ethanol, pH=3.2, flow rate 1 mL/min (for desorption). After one run of adsorption and desorption, the contents of FS and CS were increased from 3.42%, 4.30% to 23.6% and 19.4%, and the recoveries were 86.3% and 85.6%, respectively.Then used XDA-1 macroporous resin column chromatography for purifying poduct desorpted by D818 resin, the FS and CS crude mixture was received, and the contents of FS and CS were increased to 32.2% and 25.9%, and the recoveries were 85.4% and 83.3%, respectively. The process of gel column chromatography was researched as follow: eluted by petroleum ether/acetone=6:4,the flow rate 0.074cm/min,sample quality 0.012g/g gel,finally the contents of FS and CS in the product were 49.3% and 40.2%, and the recoveries were 91.3% and 92.7%, respectively.The monomers of FS and CS were detected respectively with HPLC-ELSD, IR and EIS-/MS, suggesting the chemical structure of CS and FS are unchanging after extraction, enzymolysis and purifying in enrichment process.