Studies On The Quality Control And Metabolism Of Ganoderma Lucidum And Sunrecome

Ganoderma lucidum (Leyss. ex Fr.) Karst (Polyporaceae) is a well-known Chinese herbal medicine, which has long been used in traditional Chinese medicine for the promotion of longevity and maintenance of vitality. Owing to its satisfactory clinical effect, Ganoderma lucidum has been widely used as the major component of healthy foods and drugs in China. The Shanghai Green Vally Pharmaceutical Co. cultivated G. lucidum in GAP conditions and developed an anticancer complex prescription comprised with fruit body extract and sporoderm-broken spores of G. lucidum, namely SunRecome(?). The results of pharmacological research on SunRecome(?) showed that it not only has the anticancer activity but also has the benefit of reducing chemotherapy-induced side effects. In this thesis, the quality control, metabolism of total triterpenoids in SD rats, metabolic fingerprinting were systematically studied.The chemical constituents of SunRecome(?) were systematically studied and 7 ganoderic acids were purified by silca gel, pre-TLC and pre-HPLC methods. Utilizing spectroscopic method (UV, NMR, MS), the structures of 7 compounds were fully characterized. These seven compounds were suitable as standard substances for quantitative purposes.The method of HPLC fingerprinting analysis was established for the quality control of G. lucidum and its preparation - SunRecome(?), and the LC/MS method was applied for the identification of peaks in fingerprinting chromatograms. The developed fingerprinting methods were then used to identify and access the consistency of different samples, which were collected from GAP base and different drug stores in China. The fingerprinting chromatograms have two characteristic regions. The contents of triterpenoids in region I are higher than those in region II. The six major active ganoderic acids of G. lucidum (ganoderic acids C₂, B, K, AM₁, H and D) were simultaneously quantified by HPLC/DAD method. These results showed that the content and composition of triterpenoids differed significantly in different species due to the difference in genetic source, cultivating conditions, or processing procedure. Both G. lucidum and the G. sinense are the official two species of Lingzhi recorded in Chinese Pharmacopoeia (2005 edition) and they were considered to have the same therapeutic effects. But the average content of total triterpenoids in G. lucidum was 10 times higher than that in G. sinense, which indicated that the therapeutic effects of these two species might be quite different. Therefore they are recommended to use as two different herbs in clinics.The four triterpenoids, ganoderic acids C₁, B, K and H, were qualitatively analyzed by HPLC/DAD in different bio-matrices, such as plasma, bile, urine and tissues in SD rats. The absorption, distribution, metabolism and excretion of ganoderic acids in SD rats after oral administration of G. lucidum extract were preliminarily investigated. The results indicated that the four triterpenoids were rapidly absorbed into the body fluid from the gastrointestinal tract after oral administration and widely distributed in the organs. The triterpenoids were mainly excreted from inner-body through the bile fluid.HPLC/DAD and LC/MS techniques were used to establish the metabolic fingerprinting of triterpinoids in rat serum after oral administration of G. lucidum extract, and the serum pharmacology was also applied to evaluate the pharmacological effects of serum. The mathematical statistics method was applied to analyze the relationship between metabolic fingerprinting and serum pharmacology studies. The 5 peaks in metabolic fingerprinting chromatograms have the significant relationship with the values of inhibition ratio of HeLa cell, and the 5 peaks could be chosen as the marker compound in quality control of crude drugs and their finished products.