Subcritical Fluid Extraction Of Seed Oil And Prevention Of Hyperuricemia By Flavonoid Extract From Chaenomeles Sinensis(Thouin)Koehne

Chaenomeles sinensis(Thouin)Koehne(C.sinensis)was originated in China as a processing resource for both medicine and food dual purpose.It is widely planted in China with a high yield of fruits and high percentage of seeds and therefore has broad prospect of application.In order to find a new way to develop and utilize C.sinensis,the C.sinensis seeds and fruits were investigated respectively in this paper.The results showed follows: 1.Study on methods for extraction of C.sinensis seed oil and flavonoids of C.sinensis fruitsDifferent extraction methods(ultrasonic-assisted extraction,microwave-assisted extraction,subcritical fluid extraction and cold-pressed extraction)were applied to extract oil from C.sinensis seeds.The effect of extraction methods on oil yield was investigated and the fatty acid compositions and relative contents were analyzed by GC-MS.The results showed that the order of the extraction yield: ultrasonic extraction(cyclohexane)> microwave extraction(ethyl acetate)> subcritical fluid extraction > cold-pressed extraction.Subcritical fluid extraction led to better oil quality with 88.75% unsaturated fatty acids,it's the better extraction method for extracing high quality oil from C.sinensis seeds.The extraction technics of flavonoids of C.sinensis fruits was studied by single-factor experiment.The best conditions of extraction methods were as follows: the extraction time 90 min,the concentration of ethanol 70%,material/solution rate 1:20.AB-8 is the best macroporous for purificating flavonoids of C.sinensis fruits.The optimum conditions are: the concentration of sample is 0.7mg/mL;the rate of desorption agent is 75% ethanol.The content of flavonoids is 78% after purification with AB-8 type resin(CSF),and its yield was 38.9%.The purity of ethyl acetate extraction of CSF(CSF-E)was 97%,the yields of CSF-E was 5.5%.2.Study on the extraction process of C.sinensis seed oil using subcritical fluid extraction and preparation of the O/W skin care cream with C.sinensis seed oilSubcritical fluid extraction technology was applied to the oil extraction from C.sinensis seeds.Based on the single-factor experiment and response surface methodology(Box-Behnken design)was used to optimize the extraction conditions of oil from C.sinensis seeds.The result indicated that the maximal oil yield recovery(29.56%)was obtained under the optimum conditions(35oC,35 min,3times,40mesh).The physical-chemical properties of C.sinensis seed oil extracted by SbFE is as following: acid value 0.53 mg KOH /g;iodine value 113.58 g/100g;peroxide value 0.02 mmol/kg;saponification value 185.82 mg/g,relative density 0.85.Moreover,C.sinensis seed oil was with rich in total phenolic compounds(527mg/kg),total flavonoids(15.32mg/kg)and Ve(0.957mg/kg).And the content of heavy metals was low.This part fermented C.sinensis seed oil as raw material,according to a large number of single factor experiment and after a series of orthogonal experiment to choose other components and its most appropriate amount.The result showed the best formula of the O/W skin care cream with C.sinensis seed oil is 5% C.sinensis seed oil,5% octadecanol,0.5% liquid paraffin,0.75% lanolin,2% simethicone,5% glyceryl monostearate,6% stearic acid,3% glycerinum,2.5% triethanolamine and 69.5% deionized water.3.Identification of components and evaluation of biological activity of CSF-ECSF-E was with rich in total flavonoids(973.1 mg RE/g)and total phenolic compounds(460.6 mg GAE/g).To identify the major chemical constitunents of CSF-E,we carried out HPLC-DAD-ESI-MS/MS by using the eatablished analysis method.8 flavonoids compounds are identified including B-type procyanidin dimer,B-type procyanidin trimer,neohesperidin,apigenin-7-O-glucuronide,quercitrin,ombuin and so on.Moreover,CSF-E showed excellent antioxidant activity and significantly scavenging activities on DPPH·,O2·?,OH· and H2O2 respectivity,and in concentration-dependent manners.And CSF-E showed strong xanthine oxidase inhibitory activities in vitro(IC50=207.656?g/ mL).Lineweaver-burk analysis of the enzyme kinetics data showed that the inhibition of xanthine oxidase by CSF-E was of a competitived type.4.Effects and meachanisms of CSF-E on hyperuricemic mice model by potassium oxonateThe hyperuricemia mice model was induced by intraperitoneal injection of potassium oxonate(250 mg/kg bw).The results showed the model was successful after 2h intraperitoneal injection of potassium oxonate for inducing hyperuricemia in mice.After 7-day oral administration,CSF-E at 60mg/kg,120mg/kg,180mg/kg could reduce serum uric acid,creatinine and BUN levels;serum and hepatic XOD activities were significantly suppressed compared to model group mice.Moreover,CSF-E could down-regulating renal mURAT1 and up-regulating renal mOAT1 expression to decrease urate reabsorption and enhance urate excretion.CSF-E at middle dose could cause a significant increase in the activities of SOD,GSH-PX and a significant decrease in the accumulation of MDA.These findings suggested that CSF-E produced dual hypouricemic actions to prevention and cure hyperuricemia and gout by suppressing XOD activity and regulating renal transport protein expression.