Studies On Active And Nutritional Components In Artificial Fermentation Cordyceps

Cordyceps sinensis is among the most valued medical fungi in traditional Chinese medicine which grows in the many provinces such as Sichuan, Yunnan, Qinghai, Xizang, Gansu and so on. It is commonly used in China to replenish the kidney, soothe the lung for the treatment of fatigue, night sweating, hypo-sexualities, hyperglycemia, hyperlipidemia, asthenia after sever illness, respiratory disease, renal dysfunction and renal failure, arrhythmias and other heart, and liver disease. In this thesis, an artificial fermentation Cordyceps, the substitute of Cordyceps sinensis, was chosen as the object of research, and the extraction and determination of active components in the artificial fermentation Cordyceps and its elemental speciation analysis were studied systematically, the results obtained will provide theoretical and applied foundations. The main research results and conclusions are shown as following:· Increasing the extraction percent of polysaccharide was taken as the object, different kinds of extraction technologies of polysaccharides in the artificial fermentation Cordyceps were compared, including water refluxing, microwave, ultrasonic and enzyme methods. The results showed that the extraction method with microwave was the optimum extraction technology, then it was investigated by the orthogonal optimization on microwave firepower, ratio of solid and liquid, extracting period. The optimum technical parameters were obtained: firepower 80, ratio of [S]/[L] with 1:17 for 60s. The Cordyceps polysaccharide extracted by microwave could inhibit the a-amylase activity strongly.· A method was developed for the determination of the contents of polysaccharides in Cordyceps. Polysaccharides in Cordyceps samples were extracted by water back-flowing method, after the interference components such as monosaccharide, disaccharide, oligosaccharide, polyphenol and amino acids in the samples were removed by back-flowing with 80% ethanol. The contents of the polysaccharides were determined by anthrone-H₂SO₄ colorimetry. To minimize the error of determination, a corrected factor was used. The results obtained showed that themethod used in this paper was simple and available, and the color of the treated samples could be stable within 8h.The average value of the recovery for the determination of the polysaccharides was 99.7% with the RSD of 1.0%(n=3 ).The contents of polysaccharides in the fermentation Cordyceps, Cordyceps from Xinjiang and Cordyceps from Qinghai samples were determined with the percent of 8.29%,11.90%,7.69%, respectively. The results showed there was no special difference among them on the contents of polysaccharides.? A method of reverse-phase high performance liquid chromatography (RP-HPLC) was developed for the determination of nucleotides (uridine, guanosine, adenosine and adenine) in Cordyceps. The nucleotides were well separated and determined on an Alltima C18 column (250mmx4.6mm, 5/nn) by using a mixture of acetonitrile-water (V/V=4:96) at 260nm with the flow rate of l.OmL/min. The results showed that the developed method was sensitive, accurate, reproducible and suitable for analysis of nucleotides in Cordyceps. Besides mannitol, cordycepin and ergosterol in Cordyceps were determined by HPLC, respectively. The results showed that the contents of all eight active components in the artificial fermentation Cordyceps were higher than the natural Cordyceps except of cordycepin.? The amino acids in Cordyceps were determined by HPLC. The contents of seventeen amino acids except for Ser were analyzed. The results showed that there were all of seventeen amino acids in fermentation Cordyceps. The total contents of amino acid in fermentation Cordyceps was 236.5mg/g which was lower than natural Cordyceps, with 39.7% of EAA/TAA higher than natural Cordyceps .? The elemental transference characteristics and primary speciation analysis of 18 elements in the artificial fermentation Cordyceps were first investigated by the inductively coupled plasma mass spectrometry (ICP-MS) and HPLC-ICP/MS. The elemental distribution in the artificial fermentation Cordyceps was just the same as that in the natural Cordyceps. The contents of As, Pb in the artificial fermentation Cordyceps were lower than that in the natural Cordyceps.Except that the percent of dissolved estate of P, Cr, As were higher than suspending estate, the suspending estate were the most existing form for the others. It may have something with the low water-solubility of inorganic salt and compounds which are formed as it reacts with organic molecule The polarity of extraction solvents could highly affect the solvent possibility of the analyzed elements. The transference characteristics of kinds of elements including Mg, Ca, P, Zn, As, Pb were in linearity proportion with the polarity of extraction solvents.The speciation of multi-elements were analyzed by HPLC and on-line determined with ICP-MS. The results indicated that elements were differentiated as inorganic and organic species. Inorganic species of Zn, Sr, Mn were the most species in fermentation Cordyceps, however, Al, Mg, Ca were on the contrary, and there were no distinct difference between inorganic and organic species of Ni, Cu, Fe.