Study On Isolation And Biological Activities Of Hydroxycinnamoylserotonins From Safflower Seed Meal(Carthamus Tinctorius L.)

China is rich in safflower(Carthamus tinctorius L.) seed. Safflower seed meal which is the byproduct is basically the cheap fodder for animals and its utilization is not sufficient and effective. Hydroxycinnamoylserotonins(HSs), including feruloyserotonin(FS), N-(p-coumaro- yl)serotonin(CS), N-(p-coumaroyl) serotoninmono-β-D-glucopyranoside (CSG) and N-feruloyserotonin-mono-β-D- glucopyranoside(FSG), are the active compounds in it which may be biologically active in antioxidant, preventing free radical damage and inhibiting cancer growth. Therefore isolation of HSs were necessary to study their biological activities and can extract HSs to reduce the cost and offer resource for the new and native antioxidants.First, a high performance liquid chromatography (HPLC) method was developed for determination of HSs in safflower seed meal. The results showed that the method was rapid, accurate and repeaTab, which proved feasible to determine HSs in safflower seed meal.To select the proper macroporous resin used for separating HSs in safflower oil cake, the adsorption and desorption capability of sixteen different macroporous resins in static state were studied. XDA-1 was selected as the suitable resin. The dynamic character of XDA-1 was studied and the corresponding packed column chromatography was optimized: column bed volumes 20mL, CS and FS concentration prior to application onto the column 0.276 and 0.346 mg/mL, respectively, sample size 5 bed volumes (BV), sample flow rate 6BV/hr. After one run of adsorption and desorption, the contents of FS and CS were increased from 2.04%, 1.63% to 24.7% and 22.1%, and the contents of CSG and FSG were increasd from 0.29%, 0.23% to 3.28% and 2.30%, respectively.During the experiments, the macroporous resin, C18 and preparative column chromatography were used to isolate HSs and four purier compounds were acquired. They were identified as feruloyserotonin(FS), N-(p-coumaroyl)serotonin(CS), N-(p-coumaroyl) serotoninmono-β-D-glucopyranoside (CSG) and N-feruloyserotonin-mono-β-D- glucopyranoside(FSG) by HPLC-MS-DAD, 1H-NMR and 13C-NMR.Biologically activities of HSs were studied. HSs could scavenge DPPH·, O₂^·- and·OH free radical. Their activities of scavenging radical were increased with the rising concentration and the activities of CSG and FSG were over and above FS, CS. In conclusion, the activities of scavenging radical FS, CS were more higher.The anti-tumor activities of HSs isolated from safflower oil cake were investigate in vitro by MTT assay. The results showed that HSs could inhibit the growth and had significant cytotoxicity to H446, HepG-2 and B16. The half-inhibition ratio of FS, CS were 105.0μg/mL and 173.2μg/mL on HepG-2, respectly. As a whole, the anti-tumor activities of FS and CS were more higher than CSG and FSG due to their structures. The above experiments explained HSs could be as potential nature compounds with a wide range of biologically activities to study and develop and reveal their active mechanism.