American ginseng flower polysaccharide with the efficacy of reducing weight, lipid and blood sugar, its preparation method and application

Abstract

The present invention discloses a polysaccharide from American ginseng flower with the functions of reducing weight, lipid and blood sugar, and its preparation method and application. The preparation method includes mixing the powder of American ginseng flower with an ethanol aqueous solution, heating, extracting with water as the extractant, concentrating, precipitating with ethanol, then dissolving with water, dialyzing, concentrating, drying, mixing with water, centrifuging, adding the supernatant to a DEAE Sepharose FF column, and performing gradient elution successively with pure water and 0.1 M and 0.3 M NaCl aqueous solutions, collecting the eluate obtained by gradient elution with 0.3 M NaCl aqueous solution, and drying to obtain the polysaccharide from American ginseng flower with the functions of reducing weight, lipid and blood sugar. The present invention uses the hot water extraction method to extract the polysaccharide from American ginseng flower and uses a DEAE Sepharose Fast Flow column to separate the polysaccharide from American ginseng flower. This polysaccharide has the functions of reducing cholesterol, regulating lipid metabolism, reducing blood sugar, and regulating intestinal flora.

Description

Technical Field

[0001] The present invention relates to the technical field of extracts, and particularly relates to a Panax quinquefolius flower polysaccharide with the effects of reducing weight, lipid and blood sugar, and its preparation method and application. Background Art

[0002] Obesity is a major risk factor for many cardiovascular diseases. Currently, in addition to strengthening exercise, drug treatment is also a common option, but these therapies are still challenging and sometimes cause serious adverse reactions. The gut microbiota plays an important role in host health, protecting the host from harmful microorganisms and maintaining immune homeostasis. Currently, gut dysbiosis is also considered a major hallmark of obesity and other metabolic disorders.

[0003] Panax quinquefolius is a perennial herb of the genus Panax in the Araliaceae family, native to North America, and now cultivated in the United States, China and Canada. Panax quinquefolius has been used to treat various diseases, can improve immunity, and has many pharmacological activities similar to ginseng. Currently, a large number of studies have shown that polysaccharides have activities such as immunomodulation, antioxidant, anti-tumor and anti-inflammatory. It is reported that Panax quinquefolius polysaccharide has significant anti-inflammatory effects and significant inhibitory effects on inflammatory factors. For example, Kang Zhongyu et al. disclosed a study on the optimization of the flash extraction process of Panax quinquefolius flower polysaccharide and its antioxidant activity (Kang Zhongyu, Zhao Daqing, Yao Jiajing, et al. Study on the optimization of the flash extraction process of Panax quinquefolius flower polysaccharide and its antioxidant activity [J]. Science and Technology of Food Industry, 2024, 45(7): 184-190. doi: 10.13386 / j.issn1002-0306.2023050278). They optimized the optimal extraction process of Panax quinquefolius flower polysaccharide by the response surface method, and the optimal extraction process of Panax quinquefolius flower polysaccharide was: extraction voltage: 130V, liquid-solid ratio: 30:1 mL / g, extraction time: 100s. At the same time, three verification experiments were carried out under the experimental conditions of the optimal process, and finally the average value of the polysaccharide yield was 11.12% ± 0.23%, which was close to the predicted value. In addition, Panax quinquefolius flower polysaccharide has good effects on the scavenging of DPPH free radicals, hydroxyl free radicals and total reducing power, which indicates that Panax quinquefolius flower has good antioxidant activity. However, it has not been reported that Panax quinquefolius flower polysaccharide can prevent obesity, and the interaction between polysaccharide and gut microbiota has not been disclosed. Summary of the Invention

[0004] The technical problem to be solved by the present invention is how to provide a Panax quinquefolius flower polysaccharide that can prevent obesity.

[0005] The present invention solves the above technical problem by the following technical means:

[0006] A preparation method of a Panax quinquefolius flower polysaccharide with the effects of reducing weight, lipid and blood sugar, comprising the following steps:

[0007] S1. Mix the American ginseng flower powder with an aqueous ethanol solution, heat-treat it, and then dry to obtain a pretreated powder.

[0008] S2. Use water as an extracting agent to heat-extract the pretreated powder, concentrate it, precipitate with ethanol, dissolve the obtained precipitate in water, dialyze, concentrate, and dry to obtain crude American ginseng flower polysaccharide.

[0009] S3. Mix the obtained crude American ginseng flower polysaccharide with water, centrifuge, and add the supernatant to a DEAE Sepharose Fast Flow column. Gradient elution is carried out successively using pure water, 0.1M NaCl aqueous solution, and 0.3M NaCl aqueous solution as eluents. Collect the eluate obtained by gradient elution with 0.3M NaCl aqueous solution, and dry to obtain the American ginseng flower polysaccharide with the efficacy of reducing weight, lipid, and blood sugar.

[0010] Preferably, in S1, the weight-to-volume ratio of the American ginseng flower powder to the aqueous ethanol solution is 1g:10ml; the volume fraction of ethanol in the aqueous ethanol solution is 85%; the heating temperature is 70°C; the treatment includes three treatments, and the time for each treatment is 8h; in S2, during the extraction process, the solid-to-liquid ratio is 1:10g / ml; the extraction temperature is 90°C.

[0011] Preferably, in S3, during the process of mixing the obtained crude American ginseng flower polysaccharide with water, the weight-to-volume ratio of the crude American ginseng flower polysaccharide to water is 1g:100mL; the elution flow rate is 2mL / min; the elution time for each eluent is 200min.

[0012] Preferably, in S3, it further includes mixing the product after drying the eluate with water, centrifuging to take the supernatant, using a Sepharose CL 6B column, eluting with 0.1M NaCl aqueous solution as the eluent, dialyzing, concentrating, and freeze-drying to obtain the American ginseng flower polysaccharide with the efficacy of reducing weight, lipid, and blood sugar.

[0013] Preferably, in S1 and S3, the specification of the dialysis bag used for dialysis is 3500KDa.

[0014] The present invention also provides an American ginseng flower polysaccharide with the efficacy of reducing weight, lipid, and blood sugar, which is prepared by using the preparation method of the American ginseng flower polysaccharide with the efficacy of reducing weight, lipid, and blood sugar.

[0015] Preferably, for the American ginseng flower polysaccharide with the efficacy of reducing weight, lipid, and blood sugar, its total sugar mass content is 85.60±0.55%, the uronic acid mass content is 58.30±1.18%, and the protein mass content is <0.10%.

[0016] Preferably, the American ginseng flower polysaccharide with the efficacy of reducing weight, lipid and blood sugar has a monosaccharide composition including rhamnose, arabinose, galactose, galacturonic acid and glucuronic acid.

[0017] Preferably, the molar ratio of galacturonic acid, arabinose, galactose, rhamnose and glucuronic acid is 75.56:9.39:7.93:5.32:1.80.

[0018] The present invention also provides an application of the American ginseng flower polysaccharide with the efficacy of reducing weight, lipid and blood sugar in the preparation of products for reducing weight and / or lipid and / or cholesterol and / or blood sugar and / or improving intestinal flora disorder and / or improving blood lipid metabolism.

[0019] The present invention also provides a product with the efficacy of reducing weight, lipid and blood sugar, containing the American ginseng flower polysaccharide with the efficacy of reducing weight, lipid and blood sugar.

[0020] Preferably, the product is one of food, medicine and health care products.

[0021] Taking American ginseng flower as raw material, after separating crude polysaccharide therefrom, the main component AGFPS-3 is obtained by purification through a chromatography column, and its effect on improving lipid metabolism and its influence on intestinal microorganisms of mice are explored. Subsequently, gel filtration is used to preliminarily identify the structure of AGFPS-3, providing a theoretical basis for further exploring the active ingredients in American ginseng flower and expounding the active action mechanism of American ginseng flower.

[0022] The advantages of the present invention are as follows:

[0023] (1) The present invention uses hot water extraction method to extract American ginseng flower polysaccharide, and uses DEAE Sepharose Fast Flow column to separate American ginseng flower polysaccharide. Its main component is AGFPS-3. The intervention of this polysaccharide can significantly inhibit obesity induced by high-fat diet, relieve impaired glucose tolerance, and improve the intestinal flora composition of high-fat diet mice. It has the functions of reducing cholesterol, regulating blood lipid metabolism, reducing blood sugar and regulating intestinal flora.

[0024] (2) The present invention uses Sepharose CL 6B column to purify AGFPS-3, and for the first time obtains a homogeneous polysaccharide component, purified AGFPS-3, which is a highly branched acidic heteropolysaccharide.

[0025] (3) The present invention preliminarily identifies the structure of purified AGFPS-3 through monosaccharide composition, infrared spectrum, methylation and nuclear magnetic resonance analysis, which is beneficial to further study its activity. Description of the Drawings

[0026] Figure 1Elution curve of the crude polysaccharide from American ginseng flowers prepared in Example 1 of the present invention on a DEAE Sepharose Fast Flow column;

[0027] Figure 2 Schematic diagram of the body weight changes of mice in each group fed for eight weeks in the examples of the present invention;

[0028] Figure 3 Schematic diagram of the body fat rate levels of mice in each group after eight weeks of feeding in the examples of the present invention;

[0029] Figure 4 Schematic diagram of the liver weight levels of mice in each group in the examples of the present invention;

[0030] Figure 5 Schematic diagram of the epididymal fat weight levels of mice in each group in the examples of the present invention;

[0031] Figure 6 Schematic diagram of the perirenal fat weight levels of mice in each group in the examples of the present invention;

[0032] Figure 7 Schematic diagram of the subcutaneous fat weight levels of mice in each group in the examples of the present invention;

[0033] Figure 8 Schematic diagram of the blood glucose curve of mice in each group after oral administration of glucose in the examples of the present invention;

[0034] Figure 9 Schematic diagram of the area under the blood glucose curve of mice in each group after oral administration of glucose in the examples of the present invention;

[0035] Figure 10 Schematic diagram of the serum total cholesterol (TC) levels of mice in each group in the examples of the present invention;

[0036] Figure 11 Schematic diagram of the serum triglyceride (TG) levels of mice in each group in the examples of the present invention;

[0037] Figure 12 Schematic diagram of the serum high-density lipoprotein (HDL-C) levels of mice in each group in the examples of the present invention;

[0038] Figure 13 Schematic diagram of the serum low-density lipoprotein (LDL-C) levels of mice in each group in the examples of the present invention;

[0039] Figure 14 Venn diagram based on OTU of mice in each group in the examples of the present invention;

[0040] Figure 15 Box plot of the inter-group differences of Shannon index based on OTU in the examples of the present invention;

[0041] Figure 16 This is the box plot of the difference in Chao1 index between groups based on OTU in the embodiments of the present invention;

[0042] Figure 17 This is the stacked bar chart of the relative abundances of species at the phylum level based on OTU in the embodiments of the present invention;

[0043] Figure 18 This is the stacked bar chart of the relative abundances of species at the family level based on OTU in the embodiments of the present invention;

[0044] Figure 19 This is the stacked bar chart of the relative abundances of species at the genus level based on OTU in the embodiments of the present invention;

[0045] Figure 20 This is the schematic diagram of the relative abundance levels of Firmicutes in each group of mice in the embodiments of the present invention;

[0046] Figure 21 This is the schematic diagram of the relative abundance levels of Bacteroidetes in each group of mice in the embodiments of the present invention;

[0047] Figure 22 This is the schematic diagram of the relative abundance ratio (F / B) of Firmicutes / Bacteroidetes in each group of mice in the embodiments of the present invention;

[0048] Figure 23 This is the elution curve of the prepared AGFPS-3 on a Sepharose CL 6B column in Example 1 of the present invention;

[0049] Figure 24 This is the GPC chromatogram of the purified AGFPS-3 prepared in Example 1 of the present invention;

[0050] Figure 25 This is the infrared spectrum of the purified AGFPS-3 prepared in Example 1 of the present invention;

[0051] Figure 26 This is the monosaccharide composition of the purified AGFPS-3 prepared in Example 1 of the present invention (a: monosaccharide mixed standard; b: purified AGFPS-3);

[0052] Figure 27 This is the methylation spectrum of the purified AGFPS-3 prepared in Example 1 of the present invention (a: NaBH4 reduction; b: NaBD4 reduction);

[0053] Figure 28 This is the 1H NMR spectrum of the purified AGFPS-3 prepared in the embodiments of the present invention;

[0054] Figure 29 This is the 13C NMR spectrum of the purified AGFPS-3 prepared in the embodiments of the present invention;

[0055] Figure 30 It is the heteronuclear single quantum coherence (HSQC) spectrum of the purified AGFPS-3 prepared in the embodiment of the present invention;

[0056] Figure 31 It is the correlation spectroscopy (COSY) spectrum of the purified AGFPS-3 prepared in the embodiment of the present invention. Detailed implementation manners

[0057] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of them. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0058] The test materials, reagents, etc. used in the following embodiments can be obtained from commercial channels without special instructions.

[0059] For those not specifying specific technologies or conditions in the embodiments, they can all be carried out according to the technologies or conditions described in the literature in this field or according to the product specifications.

[0060] Example 1

[0061] (1) Preparation of crude polysaccharide from Panax quinquefolius L. flowers

[0062] The dried Panax quinquefolius L. flowers were ground into powder, passed through a 60-mesh sieve. 100 g of the Panax quinquefolius L. flower powder sample was added to 1000 mL of 85% ethanol (v / v), and treated in a water bath at 70 °C for 8 h three times to remove small molecule substances such as polyphenols, pigments, monosaccharides and oligosaccharides. After the obtained sample was dried to a constant weight, it was extracted 3 times with distilled water at a solid-liquid ratio of 1:10 (w / v) g / ml, a temperature of 90 °C, and an extraction time of 3 h. The extraction solutions were combined and concentrated to one-tenth of the original volume, then three volumes of absolute ethanol were added and allowed to stand for 12 h for precipitation, centrifuged (4000 rpm, 15 min), and dissolved in distilled water. After dialysis (the specification of the dialysis bag is 3500 kDa) in ultrapure water, concentration, and freeze-drying, crude polysaccharide from Panax quinquefolius L. flowers (AGFPS) was obtained.

[0063] (2) Separation of crude polysaccharide from Panax quinquefolius L. flowers

[0064] The obtained crude American ginseng flower polysaccharide (AGFPS) was dissolved in distilled water at a solid-liquid ratio of (1:100 g / mL), and centrifuged at 5000 rpm for 5 min. The supernatant was taken and added to a DEAE Sepharose Fast Flow column (2.6×50 cm). Then, gradient elution was carried out with pure water and 0.1, 0.3, and 0.5 M NaCl aqueous solutions at a flow rate of 2 mL / min. 10 mL was collected in each tube, and 40 tubes were collected for each gradient concentration. Different components were collected and freeze-dried to obtain AGFPS-1, AGFPS-2, AGFPS-3, and AGFPS-4. The polysaccharide yields calculated by weighing were 6.54±1.28%, 5.89±1.79%, 40.26±1.92%, and 7.12±1.33% respectively. As Figure 1 shown, among them, AGFPS-3 is the main component.

[0065] (3) The separated AGFPS-3 was dissolved in distilled water at a solid-liquid ratio of (1:100 g / mL), centrifuged at 5000 rpm for 5 min, and the supernatant was taken and passed through a 0.45 μm aqueous filter membrane. A Sepharose CL 6B column (1.6×90 cm) was used, and 0.1 M NaCl aqueous solution was used as the eluent for purification at a flow rate of 0.5 mL / min. 5 mL was collected in each tube, and 40 tubes were collected. The polysaccharide components were collected by measuring A490 by the phenol-sulfuric acid method. After dialysis in ultrapure water (the specification of the dialysis bag is 3500 KDa), concentration, and freeze-drying, purified AGFPS-3 was obtained, and its elution curve is as Figure 23 shown.

[0066] Example 2

[0067] The AGFPS-3 prepared in Example 1 was used for animal experiments, which specifically included the following steps:

[0068] A total of 24 5-week-old SPF-grade male healthy C57BL / 6 mice (body weight 19.17±1.06 g) were used in this experiment. The mice were purchased from Shanghai SLAC Laboratory Animal Co., Ltd. (Shanghai, China). All mice were housed in a climate-controlled room, maintained at a relative humidity of 55%-65% and a temperature of 20-24 °C, with a light-dark cycle of 12 h:12 h. After one week of adaptive feeding with free access to food and water, the mice were randomly divided into three groups (n = 8): the NC group was fed a normal diet (10% cal from fat); the HFD group was fed a high-fat diet (60% cal from fat); the AGFPS-3 group was intragastrically administered AGFPS-3 (400 mg / kg / day) on the basis of a high-fat diet. The intervention was continuous for eight weeks. The body weight of the mice was measured and recorded once a week, and the body fat rate of the mice was measured at the end of the eighth week.

[0069] In the eighth week, the mice were fasted (for 12 - 14 h) but allowed water ad libitum on a selected day, and then intragastrically administered a glucose solution at a dose of 1.5 g / kg body weight (solution concentration: 0.4 g / mL). At 0, 30, 60, 90, and 120 min after intragastric administration of the glucose solution, the blood glucose levels of the mice were measured and recorded using a blood glucose meter, and a blood glucose - time change curve was plotted. After the test, the mice's diet was restored.

[0070] After the mice were raised for eight weeks, fresh feces were collected and immediately placed in liquid nitrogen and stored at -80 °C. Before the end of the experiment, the mice were fasted for 12 h. After the experiment ended, blood samples were collected, and the mice were sacrificed by cervical dislocation. The serum was obtained by centrifugation, and samples of liver, epididymal fat, perirenal fat, and subcutaneous fat were collected and stored in a -80 °C refrigerator. Then, mouse body weight analysis, detection of the levels of total cholesterol (TG), triglyceride (TC), high - density lipoprotein cholesterol (HDL - C), and low - density lipoprotein cholesterol (LDL - C) in the serum, non - targeted metabolism detection in the feces, and 16S rDNA amplicon sequencing analysis of the intestinal microbiota genome were performed.

[0071] The body weight parameters of the mice in each group in the eighth week of feeding are as Figure 2 shown. The body fat percentage levels of the mice in each group are as Figure 3 shown. The liver weight levels of the mice in each group are as Figure 4 shown. The epididymal fat weight levels of the mice in each group are as Figure 5 shown. The perirenal fat weights of the mice in each group are as Figure 6 shown. The subcutaneous fat weight levels of the mice in each group are as Figure 7 shown. The blood glucose curves of the mice in each group after oral administration of glucose are as Figure 8 shown. The areas under the blood glucose curves of the mice in each group after oral administration of glucose are as Figure 9 shown. The serum total cholesterol (TC) levels of the mice in each group are as Figure 10 shown. The serum triglyceride (TG) levels of the mice in each group are as Figure 11 shown. The serum high - density lipoprotein (HDL - C) levels of the mice in each group are as Figure 12 shown. The serum low - density lipoprotein (LDL - C) levels of the mice in each group are as Figure 13 shown. Figure 14 is the Venn diagram based on OTU for each group of mice. Figure 15 is the box plot of the inter - group differences in the Shannon index based on OTU. Figure 16 is the box plot of the inter - group differences in the Chao1 index based on OTU. Figure 17 is the stacked bar chart of the relative abundances of species at the phylum level based on OTU. Figure 18 is the stacked bar chart of the relative abundances of species at the family level based on OTU. Figure 19 is the stacked bar chart of the relative abundances of species at the genus level based on OTU. Figure 20Schematic diagram of the relative abundance level of Firmicutes in each group of mice. Figure 21 Schematic diagram of the relative abundance level of Bacteroidetes in each group of mice. Figure 22 Schematic diagram of the relative abundance ratio (F / B) of Firmicutes / Bacteroidetes in each group of mice.

[0072] Experimental results

[0073] It can be seen from Figure 2 that after 8 weeks of dietary intervention, the body weight of mice in the AGFPS-3 group gradually decreased, while the body weight of mice in the HFD group continued to increase. At the 8th week, the body weight of mice in the AGFPS-3 group was significantly lower than that in the HFD group. It can be seen from Figure 3 that the body fat rate of mice in the AGFPS-3 group was also significantly lower than that in the HFD group. This indicates that AGFPS-3 can inhibit the body weight gain induced by high-fat diet. It can be seen from Figures 4 - 7 that the liver weight, epididymal fat, perirenal fat and subcutaneous fat weight of mice in the AGFPS-3 group were significantly lower than those in the HFD group, indicating that AGFPS-3 treatment can effectively improve the increase in liver and adipose tissue weights induced by HFD.

[0074] As Figures 8 - 9 shown, after AGFPS-3 intervention, abnormal glucose tolerance can be improved. As Figures 10 - 13 shown, compared with the NC group, in the HFD group, the levels of TC, TG and LDL-C were significantly increased, while the level of HDL-C was significantly decreased, indicating that excessive energy intake induced lipid metabolism dysfunction, and oral administration of AGFPS-3 could significantly improve the disorder of lipid metabolism function, mainly manifested as significantly reducing the levels of TC, TG and LDL-C. Among them, oral administration of AGFPS-3 could effectively improve TG to a level similar to that of the NC group. In addition, oral administration of AGFPS-3 could also significantly increase the level of HDL-C. It can be seen that AGFPS-3 intervention can regulate the glucose tolerance and blood lipid levels of HFD mice, and has a positive effect on mouse metabolism.

[0075] AGFPS-3 improves the diversity and abundance of gut microbiota

[0076] It can be seen from Figures 14 - 16 the Venn diagram based on OTU of each group of mice that there were significant differences in the composition of the gut microbiota in the feces of the three groups of mice, and the AGFPS-3 group significantly increased the gut microbiota diversity and abundance in mice. To more specifically evaluate the specific changes in the gut microbiota structure of each experimental group of mice, the relative abundances of the microbiota in the fresh feces of each group of mice were detected at the phylum, family and genus levels. The results are as Figures 17 - 19 shown, AGFPS-3 can shift the gut microbiota structure of HFD mice towards the gut microbiota structure of NC mice. Figures 20 - 21It was shown that compared with the NC group, the relative abundance of Firmicutes in the HFD group increased significantly, while the relative abundance of Bacteroidetes decreased significantly. At the same time, as Figure 22 shown, the ratio of Firmicutes to Bacteroidetes in the HFD group was significantly higher than that in the NC group, and AGFPS-3 treatment could significantly reduce the increase in the ratio of Firmicutes to Bacteroidetes induced by HFD. This indicates that AGFPS-3 may alleviate HFD-induced intestinal flora disorder by improving the abundance of related bacteria at the phylum level, thereby regulating the health of the body.

[0077] Physicochemical properties and structural identification of purified AGFPS-3

[0078] Determination of physicochemical properties of polysaccharides: The sugar content of polysaccharide components was determined by the phenol-sulfuric acid method, the uronic acid content was determined by the m-hydroxybiphenyl method, and the protein content was determined by the Coomassie brilliant blue method. Monosaccharide composition was detected by high performance liquid chromatography. Infrared spectrum was analyzed by Fourier transform infrared spectroscopy. Methylation analysis was used. Nuclear magnetic resonance analysis was used.

[0079] The total sugar mass content of purified AGFPS-3 was 85.60 ± 0.55%, the uronic acid mass content was 58.30 ± 1.18%, and the protein mass content < 0.10%. In addition, the polyphenol showed a negative reaction when Folin-Ciocalteu reagent was used as the standard. Purified AGFPS-3 can be used for subsequent monosaccharide, methylation and NMR analysis.

[0080] As Figure 24 shown, purified AGFPS-3 obtained a single, symmetric and narrow peak by column analysis, indicating that purified AGFPS-3 is a homogeneous polysaccharide component with high purity, and its average relative molecular weight is 109.10 kDa.

[0081] Figure 25 The infrared spectrum of purified AGFPS-3 is shown. The strong and broad absorption peak at 3440 cm -1 is caused by the stretching vibration of the O-H bond, indicating strong intramolecular and intermolecular interactions between polysaccharide chains; the weak absorption band at 2924 cm -1 is caused by the stretching vibration of C-H (-CH2-); the absorption peaks at 1741 cm -1 , 1618 cm -1 represent the stretching vibrations of ester carbonyl (C=O) and carboxyl (COO-), and the weak absorption peak at 1420 cm -1 belongs to the stretching vibration of C-O (-COOH), indicating that purified AGFPS-3 is an acidic polysaccharide; the absorption peak at 1330 cm -1 corresponds to the bending vibration of C-H; the absorption peaks at 1101 cm -1 , 1019 cm -1corresponds to the stretching vibration of C-O in the pyranose ring; the α-glycosidic bond causes absorption peaks at 921 cm -1 and 833 cm -1 .

[0082] The chromatogram ( Figure 26 ) showed that the monosaccharide composition of purified AGFPS-3 was rhamnose, arabinose, galactose, galacturonic acid, and glucuronic acid. Methylation analysis was used to elucidate the glycosidic bonds of purified AGFPS-3 ( Figure 27 ). The above results indicated that purified AGFPS-3 was a highly branched heteropolysaccharide.

[0083] The configuration of purified AGFPS-3 was further elucidated by 1D (1H and 13C) and 2D (COSY and HSQC) NMR spectra ( Figures 28 - 31 ). Ten glycosidic bonds were labeled A-J for distinction (A: α-1,4-D-GalpA; B: Terminal α-D-Galp; C: β-1,4,6-D-Galp; D: β-1,3,4-D-Galp; E: Terminal α-L-Araf; F: β-1,2,4-D-Galp; G: Terminal α-L-Rhap; H: β-1,3,6-D-Galp; I: β-1,3-D-Galp; J: Terminal α-L-Arap). Figure 28 The range of the hydrogen spectrum signals in

[0084] was mainly between 4.50 - 5.30. The H signals in the ranges of 4.40 - 4.95 ppm and 4.95 - 5.30 ppm indicated the simultaneous presence of α-configuration glycosidic bonds and β-configuration glycosidic bonds in purified AGFPS-3. Among them, the signals of anomeric hydrogens mainly appeared between 4.40 - 5.30 ppm, including 5.05, 5.03, 5.01, 4.98, 4.62, 4.60, 4.57, 4.54, 4.51, and 4.48 ppm corresponding to the H-1 signals of A, B, G, E, F, I, D, H, J, and C. Among them, the anomeric carbons included 108.16, 104.70, 104.40, 104.10, 104.09, 103.79, 103.23, 100.80, 100.36, and 99.78 ppm, corresponding to E, J, H, I, C, D, F, G, A, B. The physicochemical properties of purified AGFPS-3 are as follows:

[0085] Table 1 Composition of purified AGFPS-3

[0086]

[0087] Table 2 Monosaccharide composition of purified AGFPS-3

[0088]

[0089] (GalA, galacturonic acid; Ara, arabinose; Gal, galactose; Rha, rhamnose; Glc

[0090] A, glucuronic acid)

[0091] Table 3 Results of methylation analysis of purified AGFPS-3

[0092]

[0093] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of American ginseng flower polysaccharide in the preparation of a medicament for improving lipid metabolism, characterized in that: The preparation method of the American ginseng flower polysaccharide comprises the following steps: S1. Mix the American ginseng flower powder with an ethanol aqueous solution, perform heat treatment and then dry to obtain a pretreated powder; S2. Use water as an extracting agent to heat and extract the pretreated powder, concentrate and then precipitate with ethanol, dissolve the obtained precipitate in water, dialyze, concentrate and dry to obtain the crude American ginseng flower polysaccharide; S3. Mix the obtained crude American ginseng flower polysaccharide with water, centrifuge, add the supernatant to a DEAE Sepharose FF column, and perform gradient elution successively with pure water, 0.1 M NaCl aqueous solution and 0.3 M NaCl aqueous solution as eluents, collect the eluate obtained by gradient elution with 0.3 M NaCl aqueous solution, and dry to obtain the American ginseng flower polysaccharide.

2. Use of the polysaccharide from American ginseng flower according to claim 1 in the preparation of a medicament for improving lipid metabolism, characterized in that: In S1, the weight-to-volume ratio of the American ginseng flower powder to the ethanol aqueous solution is 1 g:10 ml; the volume fraction of ethanol in the ethanol aqueous solution is 85%; the heating temperature is 70 °C; the treatment includes three treatments, and the time for each treatment is 8 h; in S2, during the extraction process, the solid-to-liquid ratio is 1:10 g / ml; the extraction temperature is 90 °C.

3. Use of the American ginseng flower polysaccharide according to claim 1 in the preparation of a medicament for improving lipid metabolism, characterized in that: In S3, during the process of mixing the obtained crude American ginseng flower polysaccharide with water, the weight-to-volume ratio of the crude American ginseng flower polysaccharide to water is 1 g:100 mL; the elution flow rate is 2 mL / min; the elution time for each eluent is 200 min.

4. Use of the American ginseng flower polysaccharide according to any one of claims 1-3 in the preparation of a medicament for improving lipid metabolism, characterized in that: In S3, it also includes mixing the product after drying the eluate with water, centrifuging to take the supernatant, using a Sepharose CL 6B column, performing elution with 0.1 M NaCl aqueous solution as the eluent, dialyzing, concentrating and freeze-drying to obtain the American ginseng flower polysaccharide.

5. Use of the American ginseng flower polysaccharide according to claim 1 in the preparation of a medicament for improving lipid metabolism, characterized in that: The total sugar mass content of the American ginseng flower polysaccharide is 85.60 ± 0.55%, the uronic acid mass content is 58.30 ± 1.18%, and the protein mass content is <0.10%.

6. Use of the American ginseng flower polysaccharide according to claim 1 in the preparation of a medicament for improving lipid metabolism, characterized in that: The monosaccharide composition of the American ginseng flower polysaccharide includes rhamnose, arabinose, galactose, galacturonic acid and glucuronic acid.

7. Use of the polysaccharide from American ginseng flower according to claim 6 in the preparation of a medicament for improving lipid metabolism, characterized in that: The molar ratio of the galacturonic acid, arabinose, galactose, rhamnose and glucuronic acid is 75.56:9.39:7.93:5.32:1.

80.

8. A medicine for improving lipid metabolism, characterized in that: Containing the American ginseng flower polysaccharide; the preparation method of the American ginseng flower polysaccharide comprises the following steps: S1. Mix the American ginseng flower powder with an ethanol aqueous solution, perform heat treatment and then dry to obtain a pretreated powder; S2. Use water as an extracting agent to heat and extract the pretreated powder, concentrate and then precipitate with ethanol, dissolve the obtained precipitate in water, dialyze, concentrate and dry to obtain the crude American ginseng flower polysaccharide; S3. Mix the obtained crude American ginseng flower polysaccharide with water, centrifuge, add the supernatant to a DEAE Sepharose FF column, and perform gradient elution successively with pure water, 0.1 M NaCl aqueous solution and 0.3 M NaCl aqueous solution as eluents, collect the eluate obtained by gradient elution with 0.3 M NaCl aqueous solution, and dry to obtain the American ginseng flower polysaccharide.

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