Yiyuankang Square In The Ganoderma Lucidum Total Polysaccharides, Total Three Terpenestudy On Extraction And Purification Process

Objective: Study Kang Yi Yuan Fang total polysaccharide Ganoderma lucidum immunomodulatory effects, the total triterpenoids, extraction, selection, isolation and purification, preferably optimal parameters, lay the foundation for further studies using cell models in vitro immunization.Methods:(1)Ganoderma lucidum polysaccharides total, total triterpenoids, Ganoderma lucidum acid A as index, collect different origins, different batches of the fungus, screened total polysaccharides, total triterpenoids, Ganoderma acid A relatively high content of Ganoderma lucidum and Ganoderma lucidum acid a, G lucidum acid Ganoderma herbs for the establishment of a reference fingerprint and cluster analysis;(2)Research total polysaccharides by orthogonal experiment and alcohol precipitation extraction process to determine the optimum process parameters; single factor experiments and uniform design to study the total triterpenes extraction process to determine the total triterpene extract optimal parameters;(3) Different types of macroporous adsorption resin adsorption properties investigated, the optimum resin, combined with orthogonal design, uniform design to determine the total polysaccharides, triterpenoids total purification process parameters and basis of single factor experiments.Results:(1)Determination of the content of Ganoderma herbs: establishing GTS, Ganoderma acid A determination method..(2)Ganoderma herbs Fingerprints: 10 batches of qualified Ganoderma herbs fingerprints, Ganoderma lucidum acid A, G lucidum acid as reference, a total of 31 common peaks calibrated total peak area of the total peak area ratio of the average of 71.77 percent; poly class analysis is divided into 10 batches of medicines Ganoderma four categories of different origin Ganoderma significant differences.(3)Ganoderma polysaccharides, extraction technology of total triterpenoids: Extraction of Ganoderma polysaccharide: 12 times the amount of water, extracted three times, each time 2h, the extraction rate could reach 83.87%; alcohol precipitation total polysaccharide technology: the relative density of 1.05 containing 80 percent alcohol content, alcohol precipitation 24 h, alcohol precipitation after mealin polysaccharide content of 25.76%; total triterpenoids extraction process: 8 times the amount of 90% ethanol, extracted three times, each time 100 min, extraction rate of 88.39 %.(4) Ganoderma polysaccharides, Purification of total triterpenoids: Select AB-8 as the best resin purification Ganoderma polysaccharides, optimized process parameters: the sample concentration of 2.0mg · ml-1, the sample solution PH = 7, the sample flow rate of 1.0ml · min-1, the sample diameter and height ratio of 1: 8, the sample volume 2.0BV; eluent of 30% ethanol, the elution amount 3BV, eluent PH = 8, elution flow rate 1.0ml · min-1, the degree of enrichment of the purified polysaccharide was 47.21% of the total increase of 23.95 percent; select HPD-400 as a purification GTS best resins, optimization of process parameters: the sample concentration 10.51 mg · ml-1, the sample solution PH = 3, the sample flow rate of 1.0ml · min-1, the sample diameter to height ratio of 1: 7, the sample volume 2.5BV; eluent selection ethyl acetate elution flow rate 1.0ml · min-1, the amount of 3.4BV, the enrichment of the total triterpenoids after purification was 54.84%.Conclusion:Through the study of Ganoderma polysaccharides, extraction and purification of total triterpenoids, the extraction and purification process is feasible to obtain total polysaccharides, stable and controllable total triterpenoids quality.