Identification Of Different Sanghuang Strains And Extraction Process And Function Of Total Flavonoids From Solid Cultures

Sanghuang is a very precious medicinal fungus in China,rich in metabolites,with antioxidant,anti-tumor,hypolipidemic and other effects.At present,most of the research on Sanghuang has focused on polysaccharides,and less attention has been paid to flavonoids.In this study,nine strains of Sanghuang were isolated and purified from wild Sanghuang fruiting bodies collected in different regions,extracted and purified from solid-state culture mycelium,and the active components and functional differences of flavonoids of different mulberry Sanghuang were compared,which provided a theoretical basis for the evaluation and screening of mulberry strain resources.The main findings are as follows:1.After isolating fresh mycelium from fruiting bodies of different sources,9 strains（S1-S9）were identified according to the results of phylogenetic analysis of strain morphology and ITS sequence.The matrix transformation ratio of flavonoids obtained by solid culture of 9 strains of Sanghuang strains was higher than that obtained in liquid culture.2.The optimal extraction conditions of Sanghuang flavonoids were determined by single factor experiment and orthogonal test as extraction temperature 80°C,extraction time:4 h,ethanol volume fraction:50%,material-liquid ratio:1:80.The flavonoid content reached 18.17 mg/g.Macroporous resin（DM301）was selected to purify flavonoids,the purification conditions were optimized,and the purification conditions were finally determined to be flavonoids at a concentration of 1 mg/m L;The flavonoid loading solution has a p H of 7;Flavonoid loading volume 35 m L;Wash volume 50 m L;The ethanol elution volume was 120 m L and the final purity of the flavonoids was 75.17%.3.Compared with the DPPH free radical scavenging ability,iron reduction ability and inhibition ofα-amylase ability of 9 Sanghuang strains flavonoids under solid-state fermentation culture.Under the same flavonoid concentration treatment,the flavonoid scavenging ability and iron reduction ability of S2,S6,S7 and S9 strains were relatively strong.The flavonoids of S1,S3,S5 and S8 strains have relatively strong ability to inhibitα-amylase.4.Through HPLC condition optimization,determine the best HPLC condition as mo bile phase:acetonitrile-formic acid water;Flow rate:1 m L/min;Column oven temperatur e:30°C;Wavelength:355 nm.The liquid phase maps of 9 strains of Sanghuang flavones were introduced into the liquid fingerprint similarity evaluation system to establish the liq uid fingerprint map,and the similarities of the 9 strains of Sanghuang flavonoids were cal culated to be 0.60,0.90,0.89,0.88,0.79,0.94,0.91,0.91,0.97,respectively.The results o f cluster analysis showed that the difference in flavonoid composition between mulberry s trains was obviously related to geographical origin and host species.5.Principal component analysis,grey correlation analysis,partial least squares regres sion analysis were used to study the spectral relationship between Sanghuang flavone com ponents and function.The results showed that when the response was DPPH radical scave nging IC₅₀,peaks 3,7,12 and 14 were the main pharmacodynamic peaks.When the respo nse is iron reducing capacity,peaks 3,7,12,and 14 are the main pharmacodynamic peaks.When the response isα-amylase IC₅₀,peaks 1,3,11,13 and 15 are the main pharmacokin etic peaks that exert the efficacy of inhibitingα-amylase.