| Kumquat which belongs to rutaceae Fortunella is a quite potential medical resource due to its rich in easy volatile, flavones, limonin and pectin, and etc. In this study, the technical conditions of Microwave Extraction and Ultrasonic Extraction on flavones and limonin were discussed. Furthermore, the activities of antioxidation on oil and DNA, free radical scavenging and bacteriostasis of kumquat crude extracts were also identified through the experiment in vitro. The whole research was to prove up the chemical constituents and biological activities of kumquat so as to provide basis for exploiting the physiological and pharmacological function of kumquat. The main study contents and results are given as follows:The volatile composition of fresh kumquat (F. Crassifolia Swingle, F. Japonica Swingle and F. margarita Swingle) separated from Liuyang, Suichuan and Gui Lin was analyzed by Solid Phase Micro Extraction (SPME) coupled with GC/MS/DS. The optimal experiment parameters for SPME were:5.0g juice,25 min at 45℃. The experiment identified forty-five volatile components in total in the three kumquat samples. It contained alkenes, alcohols, ketones, esters and etc. Most of volatile components were Alkenes.In the research of flavones which come from kumquat by Microwave Extraction, the reduced regression equation of extraction process was determined by four factors quadratic regression rotation combination design. All the tested factors affected the yield significantly, with the order as ethanol concentration> extraction time> extraction temperature> ratio of solvents and material. Predicting the model through the Aprogram solution of Excel, and according to practical situation, the optimal process conditions for Microwave Extraction were as follows:ethanol concentration 76.7%, extraction temperature 57℃, extraction time 37 min, and ratio of solvents and material 30:1. The yield of kumquat flavones was 2.181±0.007%. The retrieving rate was 98.46±1.05%, RSD=1.07%(n=5).In the study of limonin which come from kumquat by Ultrasonic Extraction, the reduced regression equation of extraction process was determined through the quadratic regression general rotation design with four factors. All the tested factors affected the yield significantly, with the order as ultrasonic treatment time> ultrasonic treatment power> soaking temperature> ratio of solvents and material. Predicting the model through the Aprogram solution of Excel, and according to practical situation, the optimal process conditions for Ultrasonic Extraction were as follows:ultrasonic treatment time 20 min, ultrasonic treatment power 440 W, soaking temperature 59℃, ratio of material and solvents 30:1, and soaking time 60 min. The yield of kumquat limonin was 3.262±0.028%. The retrieving rate was 98.03±1.55%, RSD=1.58%(n =5).Experiments on in-vitro biological activity were made using the crude extracts of flavones and limonin as materials showed the results as follows:The experimental results of study on the antioxidation effect on DNA showed that the crude extracts of flavones had antioxidation on DNA, while the test sample was at concentration of 0.16 mg/mL. When the concentration of crude extracts of limonin was more than 2.50 mg/mL, it showed that crude extracts had antioxidation on DNA.In the experiment of scavenging on free radical in vitro, the crude extracts of flavones showed scavenging effect on hydroxyl radical(·OH), superoxide anion radical (O2·) and DPPH. The scavenging order was DPPH>·OH> O2·The half scavenging concentration (IC50) were 0.45 mg/mL,0.53 mg/mL, and 41.28 mg/mL respectively. It showed that the extracts had scavenging effect on DPPH and hydroxyl radical (·OH). The IC5o were 1.76 mg/mL and 18.37 mg/mL respectively.The results of experimental study on the antioxidation effect on oil showed that kumquat crude extracts had antioxidation characteristic. The crude extracts of flavones had synergistic action with Vc, VE The crude extracts of limonin showed weak antioxidation on oil, when the dosage was 0.08%. The crude extracts of flavones had not synergistic action with VC, VE.The antibacterial experiment in vitro showed that the crude extracts of flavones had inhibitory effect on Staphyloclccus aureus, Bacillus subtilis, Excherichia coli and Mucor racemosus. The inhibiting order of the crude extracts of kumquat flavones was S. aureus> B. subtilis> E. coli> M. racemosus. The minimum inhibition concentrations (MIC) were 0.63 mg/mL,1.25 mg/mL,1.25 mg/mL and 2.50 mg/mL respectively. The half inhibiting time (IT50) was given as below:4.65 h,5.70 h,6.74 h and 9.71 h. The crude extracts of limonin also had inhibitory effect on Staphyloclccus aureus, Bacillus subtilis, Excherichia coli, and Aspergillus niger. The inhibiting order was B. subtilis> S. aureus> E. coli> A.s niger. MICs were given as below:1.25 mg/mL,2.50 mg/mL,2.50 mg/mL and 2.50 mg/mL respectively. IT50 were 4.20 h, 5.61 h,6.38 h and 7.65 h.In this paper, research was also made which used crude flavones extracts from kumquat as materials while mice was used as experimental objects, in order to discussing the effect of different locality kumquat (Liuyang and Suichuan the breed is Fortunella Crassifolia Swingle) to the physiological effects of kumquat total flavones. The main study contents and results are given as follows:Compared with model group, the experimental groups could obviously prolong the life-span of mice. The 80 mg/kg·bw doses had markness (P<0.05) effect to SOD, MDA and average life-span; the 160 mg/kg·bw doses had greatly markness (P<0.01) effect.The 20 mg/kg·bw doses of Kumquat had markness effect (P<0.05) in lowering blood lipid; and the 80 mg/kg·bw doses of Kumquat had greatly markness effect (P<0.01) in lowering blood lipid.The regulating blood sugar effect of kumquat total flavones was as below: compared with model group, after 6 days, the 160 mg/kg·bw doses had markness effect (P<0.05) to the diabetes mice's blood sugar; after 17 days, this doses had great markness effect (P<0.01). The 80 mg/kg·bw doses had markness effect (P<0.05) to the diabetes mice's blood sugar after 17 days,however it had no obvious effect to the normal mice.The effect of kumquat total flavones on the immunity system was tested, through evaluating the phagocytic function of monocyte macrophage, the ear tumefaction reduced by DNFB, swimming time, capacity of lacking of Oxygen, the index of thymus and spleen. The results were showed as below:compared with model group, the two middle doses had markness effect (P<0.05); the two higher doses had great markness effect (P<0.01).The effect of kumquat total flavones on the gastrointestinal absorption function was studied, the 80 mg/kg·bw doses had markness effect (P<0.05) to the diabetes mice's blood sugar doses had markness effect (P<0.05) while the 160 mg/kg-bw doses had great markness effect (P<0.01) on improving the gastrointestinal absorption function.
|
PDF Full Text Request