Studies On The Constituents Of Sclerotia And Fermented Mycelia Of Polyporus Umbellatus And The Quality Analysis

Polyporus umbellatus(Pers.) Fries belongs to the family of Polyporaceae,whose sclerotia are used as a folk medicine for diuresis and other kidney diseases for centuries and its bio-activities on immune system,anti-cancer and liver-protection are discovered in the past decades.Nowadays,studies on P.umbellatus are mainly focused on the polysaccharides,while our research is carried out on the minor components in order to illuminate its chemical background.Systematic isolation of the 95%alcohol extract of the sclerotia of P umbellatus on the basis of modern chromatography and methods gave twenty four compounds,twenty two of which were identified by the spectroscopic analysis(including IR,¹H-NMR, ¹³C-NMR,HMQC,HMBC,NOESY,EI-MS,FAB-MS and HREIMS ),with two new compounds and eleven isolated from the fingus for the first time.Two new ones were(20S,22R,24R)- 16,22-epoxy-3β,14α,23β,25-tetrahydroxyergost-7-en-6-one (P-2) and(23R,24R,25R)-23,26-epoxy-3β,14α,21α,22α-tetrahydroxyergost-7-en-6-one (P-4).The others were known to be ergosterol(P-1),22,23-epoxy-3β,14α,20β, 24β-tetrahydroxyergost-7-en-6-one(P-3),polyporusterone B(P-5),polyporusterone A (P-6),N-(2'-hydroxytetracosanoyl)-1,3,4-trihydroxyl-2-octodecanine(P-7),5-hydroxy-methylfurfuraldehyde (P-8),mannitol(P-9),arabinitol(P-10),α-hydroxytetracosanoic ethyl ester(P-11),friedelin(P-12),physcion(P-13),chrysophanol(P-14),ergosterol peroxide(P-15),ergosta-7,22-dien-3-one(P-16),ergosta-7,22-dien-3,5,6-triol(P-17), 2-acetyl-1,3,8-trihydroxyxanthone(P-18),nicotinic acid(P-19),uridine(P-20), adenosine(P-21),uracil(P-23). Chemical constituents of the fermented mycelia of P.umbellatus were systematically studied for the first time with variou adsordent and methods.Fifteen compounds were isolated from this material and twelve of them were identified by spectra analysis(including IR,UV,¹H-NMR,¹³C-NMR,EI-MS and FAB-MS).Apart from the compound butanedioic acid(MP-E-3),the rest were also found in the sclerotia, which were ergosterol(MP-P-1),ergosta-7,22-dien-3,5,6-triol(MP-P-2),ergosta-7,22-dien-3-one (MP-P-3),α-hydroxytetra-cosanoic ethyl ester(MP-P-4),polyporusterone B (MP-D-1),poolyporusterone A(MP-D-2),N-(2'-hydroxytetmcosanoyl)-1,3,4-trihydroxyl-2-octodecanine (MP-D-4),adenosine(MP-E-I),nicotinic acid(MP-E-2),uracil (MP-B-1),uridine(MP-B-2).Based on the studies on the chemical constituents of the selerotia of P.umbellatus, five polyporusterones isolated from the sclerotia were selected as the marker compounds, and a HPLC-DAD method was firstly developed for simultaneous quantitative determination of these five polyporusterones in the sclerotia of P.umbellatus.Varian Polaris C₁₈-A column(250 min×4.6 mm i.d.,5μm) was used.The mobile phase consisted of acetonitrile-water(28:72) with flow rate of 1.0 mL/min,and the column was kept at 30℃.The UV detective wavelength was set at 247.4 nm and the injected volume was 20μL.A HPLC fingerprint analytical method for P.umbellatus was established.Varian Polaris C₁₈-A column(250 mm×4.6 mm i.d.,5μm) was used,the mobile phase consisted of acetonitrile-water with gradient elution,with the flow rate 1.0 mL/min and UV detective wavelength 247.4 nm.The developed method can be used as an efficient meghod for quality control of P.umbellatus.Furthermore,the cultured sclerotia and fermented mycelia were analyzed compared with the wild sclerotia with this method, finding 16 common peaks in the cultured sclerotia with 10 ones in the fermented mycelia.Chemical constituents and pharmacological activities of fingi with sclerotia were viewed.