The Study Of Ganoderma Lucidum Spores Oil’s Chemical Composition And Its Antitumor Activity

Ganoderma Lucidum (Glucidum) spore oil is the lipid extracted from sporoderm-broken spores of Ganoderma lucidum by supercritical fluid with carbon dioxide.The marketing products of Glucidum spore oil are aimed at immunity-improving and anticancer effect. Pharmacological experiment study also shows that Glucidum spore has many kinds of biological activities,such as anti-cancer, immunity-improving,liver-protecting,anti-oxidant.But the levels of Terpene and polysaccharide which are functional components in Glucidum spores, are too low to make a scientific and effective evaluation of Glucidum spore oil’s quality.So It’s vital to study the active components of Glucidum spore oil for quality control of Glucidum spore oil.In this paper,we study the active components of Glucidum spore oil by it’s unsaponifiables(mainly for steroids).The main research content is divided into the following several parts:一、 The study of Glucidum spore oil’s saponification conditionsSaponification alkali concentration,saponification alkali amount,saponification time and temperature was investigated through single factor experiment.Combined with the experimental results and practical operability,determined the optimal conditions of saponification were alkali concentration lmol/L,0.22g alkali for per gram spores oil of Glucidum,saponification time1.5h,saponification temperature80℃.二、 The purification and structure identification of Glucidum spore oil’s unsaponifiable matters.8compounds was obtained through separation and purification by Silica gel column and preparative high performance liquid chromatography.By spectral analyses,physical and chemical properties and literature reported data comparison,they were identified as:(22E,24R)-ergosta-7,22-dien-3β-ol(1),ergosta-7-en-3β-ol(2),ergosta-4,6,8,(14),22-tetran-3-one(3),β-sitosterol(4),isofucosterol(5),ergosta-7,22-diene-3β,5a,6β-triol(6),ergosta-7,22-diene-3β,5a,6a-triol(7).5α,6α-epoxy-(22E,24R)-ergosta-8(14),22-diene-3p,7a-diol(8).Compound2,4,6,7are obtained from Glucidum spores oil for the first time and Compound5,8from Glucidum for the first time.三、 A preliminary study on antitumor activity of Glucidum spore oil’s unsaponifiable matters.1. The anti-tumor effect of UMGSO on S-180tumor-bearing miceThe antitumor effect of UMGSO was studied on S-180tumor-bearing mice.The result proves that UMGSO of high (700mg/kg)and medium(140mg/kg) dose can significantly inhibited tumor growth of S180tumor-bearing mice(p<0.05).Compared with model control group,the antitumor rate was45.09%、43.40%respectively,and each group has no effect on spleen index of mice.2. The effect of UMGSO on HepG2and MCF-7cell growthThe proliferation inhibition rate of UMGSO on HepG2and MCF-7cells was determined by using the MTT.The result indicates that UMGSO can notably inhibit the growth of two kinds of cells and took on dose dependently,Its IC50was14.516.8μg/mL and16.8μg/mL respectively. The above results show that UMGSO plays an important role in antitumor efficacy of Glucidum spores oil.四、 The content analysis of ergosta-4,6,8(14),22-tetraen-3-one in Glucidum spore oil.Ergosta-4,6,8(14),22-tetraen-3-one(EG) is one of the main steroidal components of UMGSO and is the particular component of fungi plant oil.What’s more,EG was effictive to various tumor cells according to present literatures,So the levels of EG in Glucidum spores oil was studied this chapter,using YMC-Pack ODS column,100%CH3OH as the mobile phase. the column temperature was300C, and the flow rate was1mL．min-1, as well as detection wavelength was350nm. We developed an HPLC method for the determination of EG in Glucidum spores oil.The results shows that the linear range of EG was L39～33.34μg-mL-1, and the average recoveries were100.70%(RSD=0.75%).The method is simple, accurate and reproducible. It can be applied to determine the contents of EG in Glucidum spores oil.