Study On The Processing Technology Of Radix Aconiti Carmichaeli Without Brine And Its Polysaccharide Extraction And Structure Characterization

Radix aconiti carmichaeli is a traditional Chinese medicinal material processed from the roots of Aconitum carmichaeli Debx.Aconite itself is a toxic medicinal material,and its clinical application must be processed to reduce its own toxicity.Traditional processing requires the addition of a large amount of brine,and the residual brine will have toxic side effects on the human body.While removing the brine residue,it will also lead to the reduction of other effective ingredients in aconite.Fuzi-polysaccharide(FPS)is one of the main active ingredients of aconite,and plant polysaccharides are currently the research hotspot of plant active substances.Therefore,research on the extraction process and structural characterization of Fuzi polysaccharide can clarify the structural basis of the active substance in aconite.In order to solve the brine residue and study the structural basis of the effective active ingredients of aconite,this research developed a brine-free aconite processing technology,and carried out experimental research on the in vitro biological activity of the brine-free aconite,and further studied the extraction process of the brine-free Fuzi polysaccharide.After optimization,the structure of Fuzi polysaccharide was characterized.The main research results are as follows:(1)The single factor effect of the processing technology of the brine-free aconite processing technology shows that when the ratio of dry raw aconite slices to the infiltration water is 1:0.4,the content of the active ingredient monoester alkaloid in the brine-free aconite is the largest,which is 0.0337%.When the drying temperature is 100℃ the content of the monoester type alkaloid of the brine-free aconite is 0.0305%.Combining the results of single-factor investigation and orthogonal experiment,the optimal processing technology for the selection of brine-free aconite is:the ratio of dry raw mulberry to infiltration water volume is 1:0.4,the steaming temperature is 120℃,the steaming time is 50 min,and the drying temperature is 100℃.At this time,the content of monoester alkaloids in the brine-free aconite is 0.3506%,and the content of diester alkaloids is 0.0148%,and it meets the requirements of the 2020 version of the Chinese Pharmacopoeia for the detection of aconite.(2)The results of the study on the in vitro biological activity of brine-free aconite have shown that the brine-free aconite has a certain in vitro antioxidant activity,and its scavenging effect on DPPH free radicals can reach 98.0%,which is much higher than that of unprocessed aconite.Brine-free aconite also has a certain in vitro hypoglycemic activity.Taking acarbose as a control,the inhibitory effect of brine-free aconite on α-amylase activity can reach 98.875%,and the inhibitory effect on α-glucosidase activity can reach 96.484%.In terms of blood lipid-lowering activity in vitro,when the concentration of brine-free aconite solution is 4 mg/mL,its binding capacity with sodium glycocholate can reach 86.66%.The influence of different concentrations of brine-free aconite liquid on the growth of intestinal probiotics Lactococcus lactis,Enterococcus faecium and Bacillus coagulans.The OD600 absorbance value of the intestinal probiotics increases first and then decreases.When cultured for 0-12 h,the microbial OD600 absorbs light The value of OD600 increased,and the OD600 absorbance value decreased when cultured for 12-28h.(3)Using ultrasonic-assisted water extraction and alcohol precipitation method,taking the extraction rate of Fuzi polysaccharide as an indicator,single factor screening and response surface analysis method were used to optimize the extraction process of Fuzi polysaccharide.When the solid-liquid ratio was 1:50,the extraction rate of Fuzi polysaccharide was 6.75%.When the ultrasound duration was 30 min,the extraction rate of Fuzi polysaccharide was 7.48%.When the extraction temperature was 90 ℃,the extraction rate of Fuzi polysaccharide was 8.33%.In addition,the extraction rate of polysaccharides obtained by alcohol precipitation with different volume fractions of ethanol is quite different,and the extraction rate of Fuzi polysaccharides obtained by alcohol precipitation with a volume fraction of 70%-90%is above 6.0%.The Box-Behnken response surface method was further used to optimize the optimal extraction process of polysaccharides from Aconite:extracting material-liquid ratio of 1:50,ultrasonic time 20 min,extraction time 2 h,alcohol precipitation concentration 80%,and the final test of the verification experiment was to test the polysaccharide extraction rate.It was 12.47%.(4)Purification and separation of the crude Fuzi polysaccharide by Sevage method and DEAE cellulose chromatography to obtain the Fuzi polysaccharide components FPS-a,FPS-b and FPS-c.Furthermore,the structure of Fuzi polysaccharide was characterized from the aspects of molecular weight,structural characteristic functional groups,glycosidic bond configuration analysis,monosaccharide composition and microstructure.The results show that:Fuzi polysaccharide contains α and β configuration of pyranose ring structure,the three components of Fuzi polysaccharide do not have a triple helix structure,and its monosaccharide composition mainly includes glucose,and a small amount of arabinose,xylose,etc.Among them,glucose accounted for a relatively high proportion of FPS-a,FPS-b and FPS-c,which were 72.6%,40.93%,and 56.65%,respectively.The weight average molecular weight of Fuzi polysaccharides is different in different components.The weight average molecular weights of FPS-a,FPS-b and FPS-c are Mw268345,Mw105007,and Mw200028,respectively.The microstructure surface of Fuzi polysaccharide is a smooth sheet with some debris;when the microscopic magnification is 3500 times,it can be observed that the surface structure of the polysaccharide is loose and there are many small particles attached to the surface.In this study,the processing technology of brine-free aconite was developed,and the anti-oxidant,hypoglycemic and blood-lipid-lowering activities of the brine-free aconite in vitro and the effect on the growth of microorganisms were studied.Furthermore,the extraction process of brine-free Fuzi polysaccharide was optimized and the structure of Fuzi polysaccharide was analyzed.This research develops a new method of aconite processing,and preliminarily analyzes the basic structure of the active ingredients in aconite,laying a solid foundation for the comprehensive development and utilization of aconite medicinal materials.