American ginseng flower polysaccharide with effects of losing weight, lowering lipid and lowering blood sugar as well as preparation method and application of American ginseng flower polysaccharide

Abstract

The invention discloses American ginseng flower polysaccharide with weight losing, lipid lowering and blood sugar lowering effects and a preparation method and application thereof.The preparation method comprises the steps that American ginseng flower powder and an ethanol water solution are mixed, water is used as an extracting agent for extraction after heating treatment, ethanol precipitation is conducted after concentration, then water is used for dissolution, dialysis, concentration and drying, and the American ginseng flower polysaccharide is mixed with water; the method comprises the following steps: dissolving American ginseng flower in water, centrifuging, adding supernate into a DEAE sepharose gel FF column, sequentially carrying out gradient elution by using pure water, 0.1 M NaCl aqueous solution and 0.3 M NaCl aqueous solution, collecting eluent obtained by carrying out gradient elution by using the 0.3 M NaCl aqueous solution, and drying to obtain American ginseng flower polysaccharide with the effects of losing weight, reducing lipid and reducing blood sugar. The American ginseng flower polysaccharide is extracted by adopting a hot water extraction method, the American ginseng flower polysaccharide is separated by adopting a DEAE Sepharose Fast Flow column, and the American ginseng flower polysaccharide has the functions of reducing cholesterol, regulating blood fat metabolism, reducing blood sugar and regulating intestinal flora.

Description

Technical Field

[0001] The invention relates to the technical field of extracts, and in particular to American ginseng flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar, and a preparation method and application thereof. Background Art

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

[0003] American ginseng is a perennial herbaceous plant of the genus Panax in the Araliaceae family. It is native to North America and is now grown in the United States, China, and Canada. American ginseng 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 immunomodulatory, antioxidant, anti-tumor and anti-inflammatory activities. It is reported that American ginseng polysaccharides have significant anti-inflammatory effects and have a significant inhibitory effect on inflammatory factors. For example, Kang Zhongyu et al. disclosed a method named "Optimization of flash extraction process of American ginseng flower polysaccharide and study of antioxidant activity" (Kang Zhongyu, Zhao Daqing, Yao Jiajing, et al. Optimization of flash extraction process of American ginseng flower polysaccharide and study of antioxidant activity [J]. Food Industry Science and Technology, 2024, 45(7): 184-190. doi: 10.13386 / j.issn1002-0306.2023050278). They optimized the optimal extraction process of flash extraction of American ginseng flower polysaccharide by response surface methodology, and obtained the optimal extraction process of American ginseng flower polysaccharide as follows: extraction voltage: 130V, liquid-to-solid ratio: 30:1mL / g, extraction time: 100s. At the same time, three verification experiments were carried out on the experimental conditions of the optimal process, and finally the average polysaccharide yield was 11.12%±0.23%, which was close to the predicted value. In addition, American ginseng flower polysaccharides have good effects on the scavenging of DPPH free radicals, hydroxyl free radicals and total reducing power, which indicates that American ginseng flowers have good antioxidant activity. However, it does not report that American ginseng flower polysaccharides can prevent obesity, nor does it disclose the interaction between polysaccharides and intestinal flora. Summary of the invention

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

[0005] The present invention solves the above technical problems through the following technical means:

[0006] A method for preparing American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar comprises the following steps:

[0007] S1, mixing American ginseng flower powder with ethanol aqueous solution, heating and drying to obtain pretreated powder;

[0008] S2, extracting the pretreated powder by heating with water as an extractant, concentrating and precipitating with ethanol, dissolving the obtained precipitate with water, dialyzing, concentrating and drying to obtain crude American ginseng flower polysaccharide;

[0009] S3. Mix the obtained American ginseng flower crude polysaccharide with water, add the supernatant to a DEAE agarose gel FF (DEAE Sepharose Fast Flow) column after centrifugation, and use pure water, 0.1M NaCl aqueous solution and 0.3M NaCl aqueous solution as eluents for gradient elution, collect the eluate obtained by gradient elution with 0.3M NaCl aqueous solution, and obtain the American ginseng flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar after drying.

[0010] Preferably, 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, each treatment lasting 8 hours; in S2, during the extraction process, the solid-liquid ratio is 1:10 g / ml; and the extraction temperature is 90°C.

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

[0012] Preferably, in S3, the process further comprises mixing the dried product of the eluate with water, centrifuging to obtain the supernatant, eluting with a Sepharose CL 6B column using a 0.1 M NaCl aqueous solution as an eluent, dialyzing, concentrating, and freeze-drying to obtain the American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids, and lowering blood sugar.

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

[0014] The present invention also provides a quinquefolius flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar, which is prepared by adopting the preparation method of the quinquefolius flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar.

[0015] Preferably, the American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar has a total sugar content of 85.60±0.55%, a uronic acid content of 58.30±1.18%, and a protein content of <0.10%.

[0016] Preferably, the American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar comprises 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 proposes the use of the American ginseng flower polysaccharide with weight loss, lipid lowering and blood sugar lowering effects in the preparation of products for weight loss and / or lipid lowering and / or cholesterol lowering and / or blood sugar lowering and / or improving intestinal flora disorder and / or improving blood lipid metabolism.

[0019] The present invention also provides a product with the effects of reducing weight, lowering blood lipids and lowering blood sugar, which contains the American ginseng flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar.

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

[0021] The present invention uses American ginseng flower as raw material, separates crude polysaccharides from it, and then purifies the main component AGFPS-3 through a chromatography column to explore its effect on improving lipid metabolism and its effect on mouse intestinal microorganisms. Subsequently, AGFPS-3 is preliminarily identified by gel molecular sieve, providing a theoretical basis for in-depth exploration of 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:

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

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

[0025] (3) The present invention conducted preliminary structural identification of the purified AGFPS-3 through monosaccharide composition, infrared spectroscopy, methylation and nuclear magnetic resonance analysis, which is conducive to further studying its activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1This is the elution curve of the crude polysaccharide of American ginseng flower prepared in Example 1 of the present invention on a DEAE Sepharose Fast Flow column;

[0027] Figure 2 This is a schematic diagram of the weight changes of mice in each group fed for eight weeks in an embodiment of the present invention;

[0028] Figure 3 This is a schematic diagram of the body fat percentage levels of mice in each group after the eighth week of feeding in the embodiment of the present invention;

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

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

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

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

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

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

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

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

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

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

[0039] Fig.14 This is a Venn diagram based on OTU for each group of mice in the embodiment of the present invention;

[0040] Fig.15 This is a box plot of the inter-group differences of the Shannon index based on OTU in an embodiment of the present invention;

[0041] Fig.16 This is a box plot of the Chao1 index group differences based on OTU in an embodiment of the present invention;

[0042] Fig.17 It is a stacked bar chart of relative abundance of species at the phylum level based on OTU in an embodiment of the present invention;

[0043] Fig.18 It is a stacked bar chart of relative abundance of species at the family level based on OTU in an embodiment of the present invention;

[0044] Fig.19 It is a stacked bar chart of relative abundance of species at the genus level based on OTU in an embodiment of the present invention;

[0045] Fig. 20 Schematic diagram of the relative abundance levels of Firmicutes in each group of mice in the embodiment of the present invention;

[0046] Fig.21 Schematic diagram of the relative abundance levels of Bacteroidetes in each group of mice in the embodiment of the present invention;

[0047] Fig. 22 Schematic diagram of the relative abundance ratio (F / B) of Firmicutes / Bacteroidetes in each group of mice in the embodiment of the present invention;

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

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

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

[0051] Fig.26 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] Fig. 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] Fig.28 is the hydrogen spectrum of the purified AGFPS-3 prepared in the example of the present invention;

[0054] Fig.29 This is the carbon spectrum of the purified AGFPS-3 prepared in the example of the present invention;

[0055] Fig.30 1 is a heteronuclear single quantum correlation (HSQC) diagram of the purified AGFPS-3 prepared in an example of the present invention;

[0056] Fig.31 This is a correlation magnetic resonance spectrum (COSY) diagram of the purified AGFPS-3 prepared in the example of the present invention. DETAILED DESCRIPTION

[0057] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0058] Unless otherwise specified, the test materials and reagents used in the following examples can be obtained from commercial sources.

[0059] If no specific techniques or conditions are specified in the examples, they can be carried out according to the techniques or conditions described in the literature in the art or according to the product instructions.

[0060] Example 1

[0061] (1) Preparation of crude polysaccharide from American ginseng flower

[0062] The dried American ginseng flower was ground into powder, passed through a 60-mesh sieve, and 100 g of the American ginseng flower powder sample was added to 1000 mL of 85% ethanol (v / v), and treated three times in a water bath at 70°C for 8 h to remove small molecules such as polyphenols, pigments, monosaccharides and oligosaccharides. The obtained sample was dried to constant weight, extracted three 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 extracted solutions were combined and concentrated to a volume of one-tenth of the original volume, and then three times the volume of anhydrous ethanol was added and kept standing for 12 h for precipitation, centrifuged (4000 rpm, 15 min), and dissolved with distilled water. After dialysis in ultrapure water (the specification of the dialysis bag was 3500 KDa), concentration, and freeze-drying, American ginseng flower crude polysaccharide (AGFPS) was obtained.

[0063] (2) Isolation of crude polysaccharides from American ginseng flowers

[0064] The obtained American ginseng flower crude polysaccharide (AGFPS) was dissolved in distilled water according to the solid-liquid ratio (1:100 g / mL), and centrifuged at 5000 rpm for 5 minutes. The supernatant was added to a DEAE Sepharose Fast Flow column (2.6×50 cm), and gradient eluted 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 obtained by weighing and calculation were 6.54±1.28%, 5.89±1.79%, 40.26±1.92% and 7.12±1.33%, respectively. Figure 1 As shown, 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, the supernatant was taken, filtered through a 0.45 μm water filter, and purified using a Sepharose CL 6B column (1.6×90 cm) with a 0.1 M NaCl aqueous solution as the eluent at a rate of 0.5 mL / min. 5 mL was collected from each tube, and 40 tubes were collected. The polysaccharide components were collected by determining A490 using the phenol-sulfuric acid method, and the purified AGFPS-3 was obtained after being dialyzed in ultrapure water (the specification of the dialysis bag was 3500 KDa), concentrated, and freeze-dried. The elution curve is shown in FIG. Fig.23 shown.

[0066] Example 2

[0067] The AGFPS-3 prepared in Example 1 was subjected to an animal experiment, specifically comprising the following steps:

[0068] A total of 24 5-week-old SPF-grade male healthy C57BL / 6 mice (weight 19.17±1.06g) were used in this experiment. The mice were purchased from Shanghai Slake 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℃, with a light-dark cycle of 12h:12h. 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 with a normal diet (10% cal from fat); the HFD group was fed with a high-fat diet (60% cal from fat); and the AGFPS-3 group was gavaged with AGFPS-3 (400mg / kg / day) on the basis of a high-fat diet. The intervention lasted for eight consecutive weeks. The body weight of the mice was measured and recorded once a week, and the body fat percentage of the mice was measured at the end of the eighth week.

[0069] In the eighth week, the mice were fasted (12-14 hours) but not watered on a selected day, and then gavaged with 1.5 g / kg body weight of glucose solution (solution concentration was 0.4 g / mL). At 0, 30, 60, 90, and 120 minutes after gavage with glucose solution, the blood glucose levels of the mice were measured and recorded using a blood glucose meter, and a blood glucose-time curve was drawn. After the test, the mice were allowed to resume their diet.

[0070] After the mice were raised for eight weeks, fresh feces were collected, immediately placed in liquid nitrogen, and stored at -80°C. The mice were fasted for 12 hours before the end of the experiment, and blood samples were collected after the end. The mice were killed by dislocation of the neck, centrifuged to obtain serum, and liver, epididymal fat, perirenal fat, and subcutaneous fat samples were collected and stored in a -80°C refrigerator. Then, the mouse weight was analyzed, serum total cholesterol (TG), triglyceride (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels were detected, non-targeted metabolic detection in feces, and 16S rDNA amplicon sequencing analysis of the intestinal microbiota genome was performed.

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

[0072] Experimental Results

[0073] Depend on Figure 2 It can be seen that after 8 weeks of dietary intervention, the weight of mice in the AGFPS-3 group gradually decreased, while the weight of mice in the HFD group continued to increase. In the 8th week, the weight of mice in the AGFPS-3 group was significantly lower than that in the HFD group. Figure 3 It can be seen that the body fat percentage 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 weight gain induced by a high-fat diet. Figures 4 to 7 It can be seen 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 HFD-induced increase in liver and adipose tissue weight.

[0074] like Figures 8-9 After AGFPS-3 intervention, abnormal glucose tolerance can be improved. Figures 10 to 13 As shown in the figure, compared with the HFD group and the NC group, the levels of TC, TG and LDL-C in the HFD group increased significantly, while the level of HDL-C decreased significantly, indicating that excessive energy intake induced lipid metabolism dysfunction, and oral administration of AGFPS-3 could significantly improve the lipid metabolism dysfunction, mainly manifested in a significant reduction in the levels of TC, TG and LDL-C. Oral administration of AGFPS-3 could effectively improve TG to a level similar to that of NC. 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 the metabolism of mice.

[0075] AGFPS-3 improves intestinal flora diversity and abundance

[0076] Depend on Figures 14 to 16 The OTU-based Venn diagram of each group of mice showed that the composition of the intestinal microbiota in the feces of the three groups of mice was significantly different, and the AGFPS-3 group significantly increased the diversity and abundance of intestinal microorganisms in the mice. In order to more specifically evaluate the specific changes in the intestinal microbial structure of each experimental group of mice, the relative abundance of the microbiota in the fresh feces of each group of mice was tested at the phylum, family, and genus levels. The results are shown in Figure 2. Figures 17 to 19 As shown, AGFPS-3 was able to shift the intestinal microbiota structure of HFD mice toward that of NC mice. Figures 20-21The results showed 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. Fig. 22 As shown in the figure, the ratio of Firmicutes to Bacteroidetes in the HFD group was significantly higher than that in the NC group, while AGFPS-3 treatment could significantly reduce the increase in the ratio of Firmicutes to Bacteroidetes induced by HFD. This suggests that AGFPS-3 may alleviate HFD-induced intestinal flora disorders by improving the abundance of bacteria at the phylum level, thereby regulating body health.

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

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

[0079] The total sugar content of the purified AGFPS-3 was 85.60±0.55%, the uronic acid content was 58.30±1.18%, and the protein content was <0.10%. In addition, polyphenols were negative when folin phenol was used as a standard. The purified AGFPS-3 can be used for subsequent monosaccharide, methylation and NMR analysis.

[0080] like Fig.24 As shown, the purified AGFPS-3 obtained a single, symmetrical and narrow peak after column analysis, which indicates that the purified AGFPS-3 is a uniform polysaccharide component with high purity, and its average relative molecular weight is 109.10 kDa.

[0081] Fig.25 The infrared spectrum of purified AGFPS-3 is shown. 3440 cm -1 The strong and broad absorption peak at 2924cm is caused by the stretching vibration of the OH bond, indicating that there are strong intramolecular and intermolecular interactions between the polysaccharide chains; -1 The weak absorption band at 1741cm is caused by the stretching vibration of CH(-CH2-); -1 、1618cm -1 The absorption peak at 1420 cm represents the stretching vibration of the ester carbonyl (C=O) and carboxyl (COO-). -1 The weak absorption peak at 1330 cm belongs to the stretching vibration of CO(-COOH), indicating that the purified AGFPS-3 is an acidic polysaccharide; -1 The absorption peak at 1101cm -1 、1019cm -1The stretching vibration of the pyran ring CO is induced by the α-glycosidic bond at 921 cm -1 and 833cm -1 The absorption peak at .

[0082] Chromatogram( Fig.26 ) showed that the monosaccharide composition of purified AGFPS-3 was rhamnose, arabinose, galactose, galacturonic acid and glucuronic acid. Methylation analysis was used to clarify the glycosidic bonds of purified AGFPS-3 ( Fig. 27 ). The above results indicate that the purified AGFPS-3 is 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 spectroscopy. Figure 28-31 ). Mark AJ to distinguish 10 glycosidic bonds (A: α-1,4-D-GalpA; B: Terminalα-D-Galp; C: β-1,4,6-D-Galp; D: β-1,3,4-D-Galp; E: Terminalα -L-Araf; F: β-1,2,4-D-Galp; G: Terminalα-L-Rhap; H: β-1,3,6-D-Galp; I: β-1,3-D-Galp; J: Terminalα-L-Arap. Fig.28 The hydrogen spectrum signal range is mainly between 4.50-5.30. The H signal in the range of 4.40-4.95ppm and 4.95-5.30ppm indicates that the purified AGFPS-3 has both α-configuration glycosidic bonds and β-configuration glycosidic bonds. The signals of anomeric hydrogen mainly appear between 4.40-5.30ppm, including 5.05, 5.03, 5.01, 4.98, 4.62, 4.60, 4.57, 4.54, 4.51 and 4.48ppm corresponding to the H-1 signals of A, B, G, E, F, I, D, H, J and C. The anomeric carbons include 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, and B.

[0084] 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 Methylation analysis results 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 the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar, characterized in that: The following steps are involved: S1, mixing American ginseng flower powder with ethanol aqueous solution, heating and drying to obtain pretreated powder; S2, extracting the pretreated powder by heating with water as an extractant, concentrating and precipitating with ethanol, dissolving the obtained precipitate with water, dialyzing, concentrating and drying to obtain crude American ginseng flower polysaccharide; S3. Mix the obtained American ginseng flower crude polysaccharide with water, add the supernatant to a DEAE agarose gel FF column after centrifugation, and perform gradient elution using pure water, 0.1 M NaCl aqueous solution and 0.3 M NaCl aqueous solution as eluents in sequence, collect the eluate obtained by gradient elution with 0.3 M NaCl aqueous solution, and obtain the American ginseng flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar after drying.

2. The method for preparing the American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar according to claim 1, 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, each treatment lasting 8 hours; in S2, during the extraction process, the solid-liquid ratio is 1:10 g / ml; and the extraction temperature is 90°C.

3. The method for preparing the American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar according to claim 1, characterized in that: In S3, in the process of mixing the obtained American ginseng flower crude polysaccharide with water, the weight volume ratio of the American ginseng flower crude polysaccharide to water is 1g:100mL; the elution flow rate is 2mL / min; and the elution time of each eluent is 200min.

4. The method for preparing the American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar according to any one of claims 1 to 3, characterized in that: In S3, the dried product of the eluate is mixed with water, centrifuged to obtain the supernatant, eluted using a Sepharose CL 6B column with a 0.1M NaCl aqueous solution as an eluent, dialyzed, concentrated, and freeze-dried to obtain the American ginseng flower polysaccharide with the effects of reducing weight, lowering blood lipids, and lowering blood sugar.

5. A polysaccharide from American ginseng flower with the effects of reducing weight, lowering blood lipids and lowering blood sugar, characterized in that: The polysaccharide is prepared by the method for preparing the American ginseng flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar as described in any one of claims 1 to 4.

6. The American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar according to claim 5, characterized in that: The total sugar content is 85.60±0.55%, the uronic acid content is 58.30±1.18%, and the protein content is <0.10%.

7. The American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar according to claim 5, characterized in that: Its monosaccharide composition includes rhamnose, arabinose, galactose, galacturonic acid and glucuronic acid.

8. The American ginseng flower polysaccharide having the effects of reducing weight, lowering blood lipids and lowering blood sugar according to claim 7, characterized in that: The molar ratio of galacturonic acid, arabinose, galactose, rhamnose and glucuronic acid is 75.56:9.39:7.93:5.32:1.

80.

9. Use of the American ginseng flower polysaccharide having weight-loss, lipid-lowering and blood sugar-lowering effects as claimed in any one of claims 5 to 8 in the preparation of products for weight-loss and / or lipid-lowering and / or cholesterol-lowering and / or blood sugar-lowering and / or improving intestinal flora disorders and / or improving blood lipid metabolism.

10. A product with the effects of reducing weight, lowering blood lipids and lowering blood sugar, characterized in that: Contains American ginseng flower polysaccharide with the effects of reducing weight, lowering blood lipids and lowering blood sugar as described in any one of claims 5 to 8.

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