| Objectives:Inonotus obliquus mainly contains polysaccharides,triterpenes,and polyphenols.triterpenes are one of the important biologically active components in Inonotus obliquus.A large number of studies have shown that triterpenes have functions such as anti-oxidation,anti-virus,anti-inflammatory,diabetes prevention and diabetes treatment.The target of this study was to extract and purify triterpenoids from Inonotus obliquus and study its antioxidant activity in vitro,which provides a certain theoretical basis and reference for the widespread use of Triterpenoids from Inonotus obliquus in natural food and medicine.Methods:(1)The content of total triterpenes was determined by UV-vis spectrophotometry.The effects of extraction solvent,reflux temperature,reflux time,ratio of material to liquid and extraction times were investigated by single factor experiments.One-factor-at-a-time design and response surface methodology(RSM)were used to determine optimal conditions for the extraction of total triterpenoid through regression analysis of the relationship between the extraction efficiency of total triterpenoid and reflux temperature,reflux time,solid-liquid ratio.(2)In order to obtain triterpenoids with high purity from Inonotus obliquus,macro-porous resin was used for purification.To establish optimized sample loading and the elution conditions,five different macroporous adsorption resins were evaluated for theiruse to purify triterpenoids from Inonotus obliquus.Macroporous resin AB-8 was selected as the most suitable one.And the influence of sample concentration,flow rate as well as eluent concentration,volume and flow rate on the adsorption and desorption efficiencies was investigated by single factor.(3)In this study,Through DPPH and ABTS free radical experiments,this paper investigates the antioxidant activity in vitro of Triterpenoids enriched by Inonotus obliquus,which provides a reference basis for the development of drugs and health foods from Inonotus obliquus.Results:(1)In this study,to optimize the reflux extraction process of total triterpenoids from Inonotus obliquus.The optimum process conditions was extraction solvent isopropanol,reflux temperature 80?,reflux time 2.5 h,ratio of material to liquid 1:22,and the extraction times once.And under the optimal process conditions,the total triterpenoid content of Inonotus obliquus was 13.15±0.076 mg/g,which was close to the theoretical value(13.12 mg/g),the fit between the actual value and the predicted value is good.(2)In this study,macro-porous resin was used for purification of triterpenoids from Inonotus obliquus.The optimal conditions determined as follows: sample concentration,0.2mg/mL,flow rate,1.0 mL/min;and 60% ethanol is used to remove impurities,15 BV of90% ethanol as eluent at a flow rate of 2.0 mL/min.Under these conditions,The purity of triterpenoids purified by AB-8 macro-porous resin was 85.13±2.658%.(3)The results showed that the crude triterpenes of Inonotus obliquus had better antioxidant activity.The anti-oxidation capacity of total triterpenes extracted from Inonotus obliquus showed a significant dose-effect relationship.Within the experimental range(0 ~1.0 mg / mL),the ability of crude triterpenes extracted from Inonotus obliquus to scavenge DPPH and ABTS free radicals increases as its concentration increases.Conclusion:In this experiment,the contents of total triterpenes were taken as the research object,and the influence of various factors on the total triterpenes content was investigated by response surface method.The optimal process was obtained,and the total triterpenes content in the Inonotus obliquus was up to 13.15±0.076 mg/g.Through the adsorption of macroporous resin to purify the crude extract of Inonotus obliquus,the optimal process was obtained.The purification rate of triterpenoids was up to 85.13 ± 2.658%.The scavenging ability of DPPH and ABTS free radicals by triterpenoids was investigated,which indicated that triterpenoids had certain antioxidant activity,and provided theoretical basis and reference value for the expansion of the research and application of Inonotus obliquus. |
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