Isolation, Purification And Structural Identification Of Organicgermanium Compounds In Germanium-enriched Ganoderma Lucidum

Ganoderma lucidum is one of high-worthy pharmaceutical fungi. Apart from its rich contents of polysaccharide, amino acids, gaoderic acid and sterol, organicgermanium is believed to be an effective component of the plant. In this work, organicgermanium compounds in germanium-enriched Ganoderma lucidum were isolated and purified by high-performance liquid chromatographyfHPLC). Pure organicgermanium compounds were obtained and their chemical structures were analysed. The goal of this experiment was to explore the form existing in germanium-enriched Ganoderma lucidum and provide some theoretical basis for researching and exploring new high-performance and low-toxicity organicgermanium anti-cancer drug.Studies on the extract method of organicgermanium compounds in germanium-enriched Ganoderma lucidum showed that the extract efficiency was different by using different extract solvent: the extract efficiency for the germanium-enriched fruit body was listed as follows: lmol/LNaOH>2mol/LNaOH>water(90癈)>95%ethanol>90%methanol, on the other hand, the extract efficiency for the germanium-enriched mycelium was : water(90癈)>2mol/LNaOH籰mol/LNaOH>90%methanol>95%ethanol. In this paper, the organicgermanium compounds was extracted from the mycelium by using water(90癈) because the mycelium contains much larger amount of organic germanium compounds than the fruit body. Following the extraction , four times volume ethanol was added in the extract solution in order to precipitate the biological macromolecular. The determination of Ge with graphite furance atomic absorption spectroscopy provided that about 50% germanium was contained in the ethanol precipitate fraction and about another 50% germanium was still retained in the fraction of water-ethanol solution. In other words, about 50% of germanium combined possibly with biological macromolecular such as polysaccharides and protein, about another 50 percent possibly combined with smallmolecular compounds such as triterpene, amino acids and peptide. Germanium-enriched mycelium and normal mycelium ofGanoderma lucidum had the similar results.Water-ethanol solution fraction was used to prepare the pure germanium compounds, five compouds were obtained through isolation, purification and collection for many times by HPLC. The results of the purity test by HPLC showed single peak respectively. In other words, these five compounds were so pure to identify their chemical structure. Through the determination of germanium content by graphite furance atomic absorption spectroscopy, five compounds contained germanium, namely, these five compounds were organicgermanium compounds.Through synthetical analysis of wave spectrum including proton nuclear magnetic resonance ('H NMR), two-dimensional (2-D) 'H-'H shift correlation NMR, two-dimensional (2-D) 'H-'3C shift correlation NMR, infrared absorption spectrum, mass specfrometry, the results were given as follows:(l)Fraction 1 was a compound whose molecular mass was 279, containingbenzene ring, carboxy, methyl, -C=C-, nitrogen and oxygen hetero atom. (2)Fraction 6 was a compound whose molecular mass was 358, containing carbonyl,-C=C-, oxygen hetero atom.(3)Fraction 8 was a compound whose molecular mass was 283, containing aldehydegroup, carboxy, -C=C-, nitrogen and oxygen hereo atom, furthermore, aldehydegroup was combined with -C=C-. This compound contained possibly cyclicstructure.(4)Fraction 9 was a compound whose molecular mass was 205, containing benzenering, carboxy and -N-H bond.(5)Fraction 10 was a compound whose molecular mass was 267, containing benzene ring, -C=C-, carbonyl .