| Aconite is one of the traditional Chinese medicines in my country.It is widely used and has the functions of regaining yang and relieving adversity,replenishing fire and helping yang,dispelling cold and relieving pain.The medicinal substance of Aconite is based on a variety of chemical components including alkaloids,flavonoids,polysaccharides,etc.currently isolated from Aconite.Among them,Aconite polysaccharide,one of the effective ingredients,has a variety of biological activities.It is manifested in the aspects of anti-tumor,anti-oxidation,immune regulation and so on.Aconite is very poisonous,and it is often used after concocting to reduce its toxicity.Heishun tablets,Baifu tablets,and Danfu tablets are common processed products of Aconite on the market.As a new type of processed product,the steamed piece formed by direct steaming of the raw piece has the characteristics of no gallbladder and no sulfur,and has become a new type of bulk commodity appearing on the market.At present,there is still a lack of research on steamed aconite and its polysaccharides.This experiment mainly conducted a preliminary study on the chemical structure of the steamed aconite polysaccharide,separation and purification,and its anti-inflammatory and antioxidant activities.The main research contents are as follows:1.Extraction and content determination of steamed aconite polysaccharideOn the basis of single factor experiments,the factor level of the steamed aconite polysaccharide extraction was determined,combined with the D-response surface method to further optimize the extraction process of steamed aconite polysaccharide,and the number of extractions(times),time(h)and material-to-liquid ratio were established(The regression model of the effect of m L/g)on??the yield(R)of polysaccharides.Finally,the optimal water extraction process conditions for distilled aconite polysaccharides were determined as follows:time(h):times:material-to-liquid ratio(m L/g)of 4:4:30.Under these conditions,the predicted polysaccharide yield is 23.22%.The phenol sulfuric acid method,the Coomassie brilliant blue method,and the carbazole sulfate method are used to determine the polysaccharide,uronic acid,and protein content in the total polysaccharides.The results show that the glucose content is 44.87%,uronic acid content is 8.47%,protein content is3.67%.Comparing the changes of the polysaccharide,uronic acid,and protein content of Aconite before and after processing,it can be known that the content of glucose,uronic acid,and protein in Aconite polysaccharide before and after processing are 16.50±1.67%,1.47±0.78%,9.80±1.56%,respectively;Steamed aconite 42.54±3.68%,8.42±1.74%,3.22±0.50%.2.Separation and purification of steamed aconite polysaccharideThe distilled aconite polysaccharide was chromatographed through anion and cation resin exchange column,and DEAE Cellulose 52 ion exchange cellulose was eluted and purified.The elution fractions were collected by the number of elution tubes-absorbance as the elution curve,and the purity was identified by high performance liquid phase,and 7 Two purified polysaccharides APS1,APS2,APS3,APY1,APY2,APY3,APY4,of which APS1,APS2,APS3 are white powder,APY1 is light yellow powder,APY2,APY3,APY4 are brown powder.3.Chemical structure analysis of steamed aconite polysaccharideThe structure of seven purified polysaccharides in the steamed aconite polysaccharides was analyzed by a combination of ultraviolet,infrared,high-performance liquid phase,UPLC,etc.The results of ultraviolet spectroscopy showed that the seven purified polysaccharides did not contain nucleic acid and protein components.The infrared spectroscopy was used to determine the structure of the seven purified polysaccharides.The results showed that the seven purified components were polysaccharide compounds.The molecular weight of APS1 is 2.48×10~4,mainly composed of D-Rib,D-Glc A,D-Gal A,D-Glc,L-Ara,D-Xyl,and its molar ratio is 0.05:0.30:0.50:31.21:0.11:0.07;APS2molecular weight is 2.01×10~6,mainly composed of D-Man,L-Rha,D-Rib,D-Glc A,D-Gal A,D-Glc,D-Gal,L-Ara,D-Xyl,Itsmolarratiois0.07:0.05:0.08:0.40:0.58:30.50:0.27:0.67:0.31;APS3 molecular weight is 1.44×10~6,mainly composed of D-Man,L-Rha,D-Rib,D-Glc A,D-Gal A,D-Glc,D-Gal,L-Ara,D-Xyl,the molar ratio of which is 0.07:0.20:0.06:0.18:0.26:36.08:0.34:1.10:0.66;APY1 molecular weight is 1.67×10~6;APY2 The molecular weight is 1.60×10~6,mainly composed of D-Man,L-Rha,D-Rib,D-Glc,D-Gal,L-Ara,D-Xyl,L-Fuc,and its molar ratio is equal to1.82:1.95:0.58:7.77:3.93:0.68:1.79:0.47;APY3 molecular weight is 1.78×10~6,mainly composed of D-Man,L-Rha,D-Glc A,D-Glc,D-Gal,L-Ara,D-Xyl,Its molar ratio is equal to 0.71:0.78:1.19:4.02:2.99:0.86:1.90;APY4 molecular weight is 1.63×10~6,mainly composed of D-Man,L-Rha,D-Rib,D-Glc,D-Gal,L-Ara,D-Xyl,L-Fuc,the molar ratio is equal to 0.47:0.78:0.78:5.32:2.41:0.38:0.78:1.11.4.Study on the anti-inflammatory activity of each component of steamed aconite and the antioxidant activity of purified polysaccharideThe carrageenan-induced rat foot swelling model was used to explore the anti-inflammatory effects of different components of steamed aconite,and to observe the rat foot swelling degree,serum IL-1β,TNF-αand PEG2 content changes to evaluate the anti-inflammatory effect of each component of steamed aconite The results show that the components of steamed aconite can significantly reduce the swelling of the feet of rats with foot swelling,the levels of IL-1β,TNF-αin the serum,and PEG2 in the foot tissues,and the effect of the decoction group is better than that of alcohol Extract and polysaccharide group.The in vitro antioxidant capacity of purified polysaccharides was evaluated by measuring the scavenging ability of DPPH free radicals and OH free radicals.The results showed that APY3 had better antioxidant capacity than other components.In this paper,a preliminary study on steamed monkshood polysaccharides laid a foundation for the research on the spatial configuration,pharmacological effects,new clinical applications and quality control of steamed monkshood polysaccharides. |
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