| Chaenomeles speciosa(Sweet) Nakai, a plant which could be used both as herb and fruit, contains abundance of biological active components. In this paper, fresh fruits was used as experimental materials. SOD was extracted from aqueous extract, oleanolic acid and flavonoids were extracted from residue comprehensively. The extraction and purification process of SOD was optimized. The extraction and purification process of both oleanolic acid and flavonoids were optimized, raw material as experimental material. Under optimized extraction process, the amount of active components from residue was compared with that extracted from raw material, thus investigating the exploitation value of residue. The conclusions were made as follows.The water content of Chaenomeles speciosa(Sweet) Nakai fresh fruit was 85.47%. The weight of raw material and dry residue occupied 14.53% and 7.71% respectively of corresponding fresh fruit.The best extraction conditions of SOD: water as solvent, with 1:2 material-liquid ratio, extracting for 1h. The seperation and purification of SOD contains thermal denaturation, ultrafiltration, acetone precipitation and Sephadex G100 column chromatography. The absorption peak of SOD product was at 276 nm, and H2O2 inhibited the very likely being Fe-SOD. Raw material of Chaenomeles speciosa(Sweet) Nakai with granularity 60 was used as experimental material, and ethe optimized extraction process of oleanolic acid was confirmed: ethanol as solvent, 1:20 material-liquid ratio, 60 ℃ for 20 min under ultrasonic power of 200 W using ultrasonic-assisted extraction technology. The optimized extraction process of total flavonoids was confirmed: 70% ethanol as solvent, 1:20 material-liquid ratio, 50 ℃ for 20 min under ultrasonic power of 200 W using ultrasonic-assisted extraction technology. ADS-17 showed a better performance in adsorption and desorption properties. Optimal adsorption of flavonoids was achieved with the sample flavonoids concentration of 2.04 mg/ml, 2 ml/min flow rate and eluting with 30% and 50% ethanol, 1.5 ml/min flow rate, 4 BV and 3 BV in volume respectively. The flavonoids products was obtained from 30% and 50% ethanol eluent.The comprehensive extraction process of active components from Chaenomeles speciosa(Sweet) Nakai was confirmed: SOD was extracted from juice; oleanolic acid was extracted from dry residue using ethanol as extraction solvent; quadratic residue was remained, from which total flavonoids were extracted using 70% ethanol. Processes above were all under optimum extraction processes. The specific activity of SOD from Chaenomeles speciosa(Sweet) Nakai was 689.98U/mgpr, or 422.61U/g fresh fruit. The recovery ratio of total SOD activity was 7.89%, purification degree was 17.36 through purification process. The amounts of oleanolic acid in dry residue and raw material were 1.63% and 1.46% respectively. Oleanolic acid crystal was obtained after purification of crude extraction solution. Purity of crystal from raw material and dry residue were both above 90% under HPLC. The amounts of flavonoids in dry residue and raw material were 7.63% and 8.64% respectively. Flavonoids product was obtained after macroporous resin purification, with yield between 1.5% and 2%, in purity of about 95%. SOD of 42261 U, oleanolic acid of 125.67 mg and flavonoids of 588.27 mg could be obtained from every 100 g fresh fruit in the comprehensive extraction process.In this study, extraction and utilization of multiple objective active components were explored and proved to be practicable. Considerable active components were remained in processing residue of Chaenomeles speciosa(Sweet) Nakai, showing high exploitation value, thus providing theoretical supports to utilization of residue abandoned after deep processing in industrial production. |
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