| Diabetes has become the third most serious threat to human health in noncommunicable diseases.Common hypoglycemic drugs were generally synthetic,which effects were significant but side effects were also very obvious with long-term use,so more and more scholars tended to search natural plant resources with hypoglycemic effect.Anoectochilus was the famous Chinese herbal medicine,which mainly was used for the treatment of diabetes,nephritis embolism in folk.However,the study of Anoectochilus was still in its infancy,its development and utilization were far from enough.The main study contents of this paper were as follow:(1)Using the detection method of the national standard of the People’s Republic of China,the medicinal parts of Anoectochilus were determined and the relative content of physiological actives were analyzed,providing the basic data for the development and utilization of Anoectochilus.(2)The physiological active of Anoectochilus extract was studied,including the half inhibitory rate of α-glucosidase,the mechanism of inhibition of α-glucosidase and the antioxidant capability in vitro with the method of the DPPH·,·OH and O2-· eliminating experiments.(3)The composition and material structure of hypoglycemic actives of Anoectochilus were analyzed preliminarily with the use of advanced spectral instruments,and the relationship between microcosmic chemical composition,material structure and macroscopic hypoglycemic capability was studied.(4)The optimal extraction method and extraction conditions of hypoglycemic actives were studied by using the inhibition rate of α-glucosidase activity as an index,and the extraction process was optimized.(5)The quality evaluation system of Anoectochilus was established in order to optimize the variety and the cultivation method of Anoectochilus.(6)The scheme of recycling of Anoectochilus resources was designed which provided an alternative route for the recycling of Anoectochilus from the perspective of process to achieve the high value utilization of Anoectochilus resourcesThe main conclusions of this paper were as follow:(1)The results showed that the main components of the dry Anoectochilus are as following: ash 10.52%,reducing sugar 15.5%,polysaccharide 7.18%,protein 16.94%,flavonoids 1.61%,total dietary fiber 44.76%,the other 3.49%,which showed that Anoectochilus was with a large number of active ingredients and was worth in-depth development and utilization.(2)The hypoglycemic physiological capability and the antioxidant ability of Anoectochilus extract were studied in vitro.The results showed that Anoectochilus extract possessed an inhibitory capability against α-glucosidase and the usage amount of extract is 19.01 mg with the inhibition rate of α-glucosidase reaching 50%.The inhibitory mechanism of α-glucosidase was determined by the kinetic curve,which was mixed competitive inhibition mechanism,and the mechanism of noncompetitive inhibition was dominant.Anoectochilus extract possessed antioxidant capability and the usage amount of extract is 5.6mg,7.4mg,4.5mg with the elimination rate of DPPH·,·OH,O2-· reaching 50%.However,the results of parallel experiment showed that the physiological capability of extract was only 2.5% of the ability of acarbose,and the antioxidant capability was only 4%,7% and 8% of the ability of Vc against ·OH,DPPH· and O2-·.(3)The results showed that there were at least 15 kinds of components in Anoectochilus extract by the use of UPLC-MS.Two of the higher proportions are oleanolic acid and quercetin-3-O-glucoside.The molecular structure of quercetin-3-O-glucoside contained a large number of phenolic hydroxyl groups,which are recognized as radical inhibitors.The infrared spectrum of Anoectochilus extract was also confirmed the existence of polyhydroxy structures.These were consistent with the results of the evaluation of the antioxidant capability of Anoectochilus extract.(4)The optimal extraction method and extraction conditions of hypoglycemic actives were studied by using the inhibition rate of α-glucosidase activity as an index,and the extraction process was optimized.The results showed that the best extracting reagent was water and the optimum extraction temperature,extraction time,extraction solid-liquid ratio and extraction times were 60℃,90 min,1:15(g:mL)and 1.The results showed that extraction temperature had the greatest effect on the extraction efficiency,the extraction time was the second,and the solid-liquid ratio almost had little effect.Under these conditions,the inhibition rate of α-glucosidase reached 32.3%.(5)According to the quality evaluation system of Anoectochilus established in this paper,the inhibition rate of α-glucosidase and the antioxidant capability were used as indexes to optimize the varieties and cultivation methods of Anoectochilus to control the quality of the medicinal herbs and achieve the GAP specification management.Based on the results of four cultivars and three ways of the cultivation of Anoectochilus,from the perspective of improving hypoglycemic capability and antioxidant capability,the optimized cultivation scheme of Anoectochilus was as follows: in the sterile bottle breeding room for 6 months,and then transplanted to the forest to grow 8 months.Under this condition,the inhibitory rate of α-glucosidase reached more than 30%,and the comprehensive index of antioxidant was about 23% higher than that of the bottle.(6)The results showed that there was a positive correlation between the two data,the data of the inhibition rate of α-glucosidase and the data of polysaccharide content,while the inhibition rate did not show a positive relationship with the flavonoid content.It could be primarily proved that the inhibitory effect of Anoectochilus on α-glucosidase was mainly caused by polysaccharides.However,the inhibitory capability was not consistent with its antioxidant ability.In other words,the former high,the latter was not necessarily high,and the two physiological capabilities were independent of each other.(7)In this paper,the design scheme of recycling of Anoectochilus was put forward and verified experimentally.With the method of 100 g Anoectochilus(dry matter)as raw material,25.86 g of hypoglycemic extract,0.87 g of flavonoid extract and 39.65 g of dietary fiber were obtained.The hypoglycemic actives extraction yield was 25.86% and the inhibitory rate of α-glucosidase was 29.5%.The utilization rate of dietary fiber was 88.6% and the application performance indexes were as follow: the water retention capacity 7.68g/g,the oil holding capability 4.51g/g,the expansive force 6.12 m L/g,which were doubled better than wheat bran and were not far behind soybeans dietary fiber.The innovations of this paper were as follow:(1)From the perspective of in vitro physiological capability evaluation to study the medicinal ingredients of Anoectochilus,the relationship between structure and performance was primarily established.(2)Using the inhibition rate of α-glucosidase as an index,the extraction method and extraction conditions of hypoglycemic actives from Anoectochilus were optimized.(3)In order to optimize the variety and the cultivation method of Anoectochilus,the quality evaluation system of Anoectochilus was established. |
