Extraction Of Cordyceps Sinensis Matrix Polysaccharide And Its Application In Tobacco Liqui

China is the world’s largest producer and consumer of tobacco.The upper leaves account for about 40%of the total tobacco production in the country.However,the quality of upper leaves is generally low,characterized by strong pungency,heavy off-flavour,and insufficient aroma.Due to the lower utility of upper leaves in cigarette formulations,there has been a buildup and waste of these leaves.Enhancing the usability of upper leaves has become a focal point in the tobacco industry.Moisturizers and flavorings,as key additives,effectively enhance the quality of tobacco leaves by improving the smoking experience and modifying the aromatic style.In this study,polysaccharides were extracted from the Cordyceps cicadae culture medium culture medium.These polysaccharides were then applied as moisturizers sprayed onto the tobacco leaves,resulting in an improvement in the smoking quality of the leaves.However,their impact on aroma enhancement was not significant.Flavoring were prepared using the Maillard reaction.The polysaccharides were mixed with the flavoring and sprayed onto the upper tobacco leaves.This treatment led to improvements in the aroma,pungency,and smoking experience of the leaves,with a significant increase in the content of volatile compounds responsible for the aroma.The specific details of the study are presented below.1.Extraction and preliminary identification of polysaccharides from Cordyceps cicadae culture medium.Firstly,through the design of single-factor experiments and orthogonal optimization experiments involving different ultrasonic times,solid-liquid ratios,extraction times,extraction temperatures,and grinding degrees,the optimal process parameters for extracting polysaccharides from the Cordyceps cicadae culture medium were determined.The best conditions were found to be a temperature of 80°C,mesh size of 100,and a solid-liquid ratio of 1:25.Under these conditions,the extraction yield of polysaccharides was 1.39%.Secondly,the crude polysaccharides were subjected to separation and purification using the Sevag protein removal method,DEAE-52 column chromatography,and cellulose dialysis membrane.This process primarily yielded two types of polysaccharides,CP-1 and CP-2.Finally,the purity of the polysaccharides was assessed using ultraviolet spectroscopy,and the results indicated that both CP-1 and CP-2 were free from proteins and nucleic acids.Furthermore,FTIR was employed for preliminary identification of the polysaccharide structures,confirming that the polysaccharides from the Cordyceps cicadae culture medium exhibit the fundamental characteristics of polysaccharides.2.Preparation of perfum.Firstly,sensory evaluations were conducted through a single-factor experiment to assess the impact of sugars and amino acids on the quality of tobacco leaves.Based on the scores obtained,fructose and arginine were chosen as substrates for the preparation of flavorings.Using the fructose-arginine system,flavorings were prepared using the Maillard reaction.As the carbonyl-to-amino ratio,heating temperature,and heating time increased,the degree of browning and the content of MRPs（Maillard reaction products）gradually increased.Subsequently,an orthogonal optimization experiment was designed to determine the optimal reaction conditions,with the content of MRPs measured by OD₂₉₄ as the indicator.The best reaction conditions were determined to be a carbonyl-to-amino ratio of 2.5:1,a heating temperature of 120°C,and a heating time of 150 minutes.3.Exploration of conditions for polysaccharide fermentation of tobacco leaves.This study explored the fermentation temperature,additive ratio,and fermentation time as three factors affecting the fermentation of polysaccharides in tobacco leaves.Within the temperature range of 25-40°C,an increase in temperature gradually worsened the pungency and aftertaste of the tobacco leaves.Within the range of 5-15‰for the polysaccharide additive ratio,an increase in the ratio resulted in a gradual reduction of pungency and volatile gases in the upper leaves.During the fermentation period of 0-60 days,fermenting the upper leaves for 30 days improved the aftertaste,pungency,volatile gases,and aroma quality.Based on the overall sensory evaluation scores,the optimal conditions were determined as follows:fermentation temperature of30°C,additive ratio of 15‰,and fermentation time of 30 days.4.Analysis of chemical components and volatile substances in tobacco leavesThere were no significant differences observed among the tobacco leaves from different treatments in terms of starch content,nicotine content,and disaccharide differential.However,the application of polysaccharides and mixed liquid significantly improved the sugar content proportion in the upper leaves and increased the sugar-to-nicotine ratio.The increase in the sugar-to-nicotine ratio led to an improvement in the sensory evaluation of the upper leaves.The HS-SPME-GC-MS method was used to detect differentially volatile substances in tobacco leaves,and a total of 271 volatile substances were detected.224 volatile compounds were detected in the CK group,218volatile compounds were detected in the GDT group,and 232 volatile compounds were detected in the HH group.Among them,there are 12 types of differential substance between the CK and GDT groups,and 15 types of differential substance between the CK and HH groups,accounting for 4.42%and 5.53%of the total volatile components,respectively.There was a significant increase in the content of aroma inducing substances in the HH group’s up-regulated differential substance,and the relative content of Megastigmatrienone,Benzaldehyde,Phenylethyl Alcohol,Butyl propionate,as typical aroma inducing substances,was significantly or extremely significantly increased.