Use of elaeagnus umbellate polysaccharide in preparation of medicine for treating intestinal barrier related diseases

Abstract

The application provides application of Elaeagnus conferta polysaccharide in preparation of a medicine for treating intestinal barrier related diseases, and relates to the technical field of natural medicine chemistry. The intestinal protection effect of the Elaeagnus conferta polysaccharide on a dextran sodium sulfate induced intestinal injury model of mice is investigated, and the research result shows that the Elaeagnus conferta polysaccharide can treat clinical symptoms such as diarrhea, enteritis and weight loss caused by intestinal barrier injury, significantly reduces disease scores, inhibits colonic tissue goblet cell necrosis, promotes secretion of mucin, increases expression of tight junction proteins, and enhances intestinal mucosal barrier function. As a widely planted fruit and a fruit loved by the public, the Elaeagnus conferta has the advantages of high safety and easy acquisition. It is expected to become an effective active ingredient source for resisting intestinal injury diseases, and has a broad market development prospect.

Description

Technical Field

[0001] This invention relates to the field of natural medicinal chemistry, and in particular to the application of goat milk fruit polysaccharide in the preparation of drugs for treating intestinal barrier-related diseases. Background Technology

[0002] With socio-economic development, modern people no longer need to worry about food scarcity. However, new problems have arisen. The overuse of food additives and irregular eating habits have made people's intestines fragile and vulnerable. These unhealthy eating habits irritate the gastrointestinal mucosa, causing most people to suffer from gastritis of varying degrees. As the body's "second brain," the health of the digestive system directly determines people's quality of life. Maintaining intestinal health is a major challenge facing modern people.

[0003] The gut is a crucial interface between the body and the external environment, responsible not only for nutrient absorption but also for defending against toxins and microbial invasion. Therefore, the integrity of the intestinal mucosal barrier is a key indicator of gut health. The intestinal mucosal barrier, composed of gut microbiota, mucus layer, antimicrobial molecules, intestinal epithelial cells, and immune cells, is the "first line of defense" preventing pathogens from contacting the intestinal epithelium and is essential for maintaining intestinal homeostasis. When the barrier is damaged, bacteria and endotoxins in the gut are more likely to translocate, leading to enterogenic infections and even inducing systemic inflammatory responses and multiple organ failure. Studies have shown that damage to the intestinal mucosal barrier can participate in the pathological processes of various diseases, including inflammatory bowel disease, colorectal cancer, diabetes, non-alcoholic fatty liver disease, obesity, and stroke, through the brain-gut axis and liver-gut axis. Protecting the intestinal mucosal barrier helps maintain the body's internal environment and reduces the incidence of disease.

[0004] Currently, clinical treatment options for diseases related to intestinal barrier damage are limited, primarily focusing on symptomatic relief. This includes using aminosalicylic acid derivatives, glucocorticoids, and immunosuppressants to control inflammatory responses, or using probiotics to regulate gut microbiota balance. However, these treatments generally have several drawbacks: aminosalicylic acid derivatives have limited efficacy and are prone to drug resistance with long-term use; glucocorticoids and immunosuppressants have significant side effects, potentially leading to serious adverse reactions such as osteoporosis, liver and kidney damage, and weakened immune function; the efficacy of probiotic preparations is greatly affected by the strain, dosage, and individual patient differences, and their mechanisms of action are relatively simple, making it difficult to fundamentally repair the damaged intestinal barrier. Therefore, developing a novel therapeutic drug that is safe, effective, has few side effects, and can significantly repair intestinal barrier function has become a research hotspot and urgent need in the biomedical field.

[0005] Goat milk fruit ( Elaeagnus sarmentosa*Rehd.*, belonging to the Elaeagnaceae family, is used medicinally for its roots, fruits, and leaves. Other names include Milk Pepper, Cow Lice Fruit, Sheep Mountain Pepper Tree, Long-creeping Elaeagnus pungens, and Goat's Milk Head. It is mainly distributed in Yunnan, Guangxi, Guangdong, Sichuan (Bazhong), Hubei, and southern Henan. It has a sour taste and its main functions include relieving cough and asthma, and astringing and stopping diarrhea. It is used to treat asthma, chronic cough, chronic bronchitis, traumatic swelling and pain, rheumatic pain, and jaundice. It can also treat hematemesis, hemoptysis, sore throat, colds, infantile convulsions, and sores. *Rehd.* is rich in sugars. Studies have shown that polysaccharides are biologically active substances that can activate immune cells, improve the body's immune function, and some also have anti-aging and antioxidant effects, making it a promising product in the health food market.

[0006] However, to date, there have been no reports on the role of goat milk fruit polysaccharide in intestinal barrier protection, and no research or technical solutions have been disclosed regarding its application in the preparation of drugs for treating diseases related to intestinal barrier damage.

[0007] In addition, traditional polysaccharide extraction processes have significant drawbacks. For example, traditional water extraction and alcohol precipitation have low extraction rates. The three-phase partitioning (TPP) extraction process currently used typically employs tert-butanol, which is volatile, flammable, and toxic. Therefore, it is also necessary to optimize and improve it. Summary of the Invention

[0008] In view of this, the purpose of the present invention is to provide an application of goat milk fruit polysaccharide in the preparation of a drug for treating intestinal barrier-related diseases, so as to solve the problems existing in the prior art.

[0009] To achieve the above objectives, the technical solution of the present invention is as follows: In a first aspect, the present invention provides the use of goat milk fruit polysaccharide in the preparation of a medicament for treating intestinal barrier-related diseases.

[0010] Furthermore, the intestinal barrier-related diseases include intestinal damage.

[0011] Furthermore, the symptoms of the intestinal injury include at least one of weight loss, loose stools, fecal occult blood, shortened colon, goblet cell necrosis of colonic tissue, and mucosal edema and sloughing.

[0012] In a second aspect, the present invention provides a method for preparing the above-described *Acer buergerianum* polysaccharide, the process comprising homogenizing *Acer buergerianum* and then freeze-drying it to obtain *Acer buergerianum* powder. The *Acer buergerianum* powder is then extracted in a three-phase distribution system containing an organic solvent and a salting-out solution under a set extraction temperature and ultrasonic assistance. After extraction, the aqueous phase is concentrated and dried to obtain *Acer buergerianum* polysaccharide. The organic solvent used is a eutectic solvent, and the salting-out solution is an aqueous solution of ammonium sulfate.

[0013] The three-phase partitioning method (TPP) involves adding inorganic salts and organic solvents to the original extract, resulting in the formation of three distinct phases under various conditions. The top phase is an organic phase, primarily composed of low-polarity substances such as lipids; the middle layer is a protein layer; and the bottom phase is an aqueous phase, mainly containing water-soluble substances such as polysaccharides. This method is characterized by its simplicity, low cost, and rapid operation.

[0014] Deep eutectic solvents (DES) are solutions of ionic complexes formed by hydrogen bond acceptors and hydrogen bond donors, with freezing points significantly lower than any of their constituent components. DES is an economical and environmentally friendly solvent due to its simple preparation, low cost, and biodegradability. Currently, DES is widely used in biomedicine, organic synthesis, and food safety testing, and its development prospects are promising.

[0015] Compared to traditional extraction and separation techniques, TPP technology has many advantages, but it also has some shortcomings. The main problem is that the TPP system typically uses tert-butanol, which is highly volatile, flammable, and toxic. This invention utilizes DES instead of tert-butanol, which not only solves the pollution and toxicity problems of the original system, but also improves the extraction rate of polysaccharides.

[0016] Furthermore, in the eutectic solvent used, the hydrogen bond acceptor is selected from any one of decanoic acid, terpineol, and lactic acid, and the hydrogen bond donor is selected from any one of lauric acid, betaine, and glucose.

[0017] Furthermore, the molar ratio of hydrogen bond donor to hydrogen bond acceptor is 1:1 to 2:1.

[0018] Furthermore, the hydrogen bond donor is lauric acid, the hydrogen bond acceptor is decanoic acid, and the molar ratio of lauric acid to decanoic acid in the system is 1:1.

[0019] Furthermore, the mass fraction of ammonium sulfate in the aqueous solution of ammonium sulfate used is 15-30%.

[0020] Furthermore, the moisture content of the DES used is 15%–35%; Furthermore, the ratio of DES to ammonium sulfate solution (volume ratio of upper and lower phases) used is 0.5:1 to 2.5:1; The extraction temperature is 40℃~80℃, and the ultrasonic extraction time is 40min~80min.

[0021] Furthermore, the extraction parameters of the method are as follows: extraction temperature 60℃, ultrasonic time 60min, organic solvent is a mixture of lauric acid and decanoic acid in a molar ratio of 1:1 (water content is 30%); salting-out solution is an ammonium sulfate aqueous solution with a mass fraction of 25%; and the volume ratio of organic solvent to salting-out solution is 2:1.

[0022] It should be noted that the goat milk fruit polysaccharide in the above-mentioned applications can also be obtained by the traditional water extraction and alcohol precipitation method, that is, by extracting with hot water and then precipitating with alcohol to obtain goat milk fruit polysaccharide. This method is well known in the industry and will not be described in detail here.

[0023] In the above-described applications of the present invention, the goat milk fruit polysaccharide obtained by the extraction process provided by the present invention is preferred.

[0024] In a third aspect, the present invention provides a structure of a goat milk fruit polysaccharide used in the above-described applications, wherein the monosaccharide composition of the goat milk fruit polysaccharide comprises, in molar ratio, 35-40% galacturonic acid, 15-20% glucose, 15-20% galactose and 10-15% arabinose; and the molecular weight of the goat milk fruit polysaccharide is 8.0-535 kDa.

[0025] Preferably, the monosaccharide composition of the goat milk fruit polysaccharide includes hemiuronic acid (38.77%), glucose (18.43%), galactose (16.52%) and arabinose (12.71%), with a molecular weight of 2.15 kDa.

[0026] Furthermore, the drug contains goat milk fruit polysaccharide as an active ingredient, and also contains pharmaceutically acceptable excipients; the dosage form of the drug is solid, liquid or semi-solid.

[0027] In this invention, in vivo activity experiments were used to investigate the protective activity of goat milk fruit polysaccharide solution against the intestinal barrier.

[0028] An animal model of intestinal injury was established by gavage with sodium dextran sulfate and a goat milk fructose solution was administered. The results showed that goat milk fructose had a good therapeutic effect on intestinal injury induced by sodium dextran sulfate, with higher doses showing better effects and exhibiting a certain dose dependence. Specifically, (1) goat milk fructose could improve the clinical symptoms of loose stools, weight loss, and colonic atrophy caused by intestinal injury in mice and reduce disease scores; (2) goat milk fructose could inhibit goblet cell necrosis and mucosal edema and shedding in colonic tissue and enhance the intestinal mucosal barrier function. The above research results indicate that goat milk fructose can play an important role in the preparation of preparations that protect the intestinal barrier.

[0029] The beneficial effects of this invention include at least the following: (1) This invention verifies through a mouse intestinal injury model induced by dextran sulfate sodium that goat milk fructose can effectively improve clinical symptoms such as loose stools, weight loss, and colonic atrophy caused by intestinal barrier damage, significantly reduce disease activity scores, and show dose-dependent effects, with higher doses showing better effects.

[0030] Its mechanism of action is clear: it can inhibit goblet cell necrosis and mucosal edema and shedding in colonic tissue, and maintain the integrity of the intestinal mucus barrier by upregulating mucin (MUC2) expression and promoting acidic mucin secretion; at the same time, it can regulate the balance of inflammatory factors, reduce the levels of pro-inflammatory factors such as TNF-α and IL-1β, and increase the content of anti-inflammatory factor IL-10; it can also improve colonic oxidative stress, increase SOD and GSH activity, reduce MPO and MDA levels, and upregulate the expression of tight junction proteins such as ZO-1 and Occludin, comprehensively repairing the damaged intestinal mucosal barrier. Its therapeutic effect is superior to that of the positive control drug mesalazine, providing a highly effective treatment direction for diseases related to intestinal barrier damage.

[0031] (2) The optimized method for preparing goat milk fruit polysaccharide of the present invention uses a low eutectic solvent instead of tert-butanol, avoiding the use of toxic and harmful reagents in traditional processes, and has the advantages of being non-toxic, pollution-free, and recyclable. This process has excellent extraction efficiency, with a goat milk fruit polysaccharide extraction rate of 54%, providing a high-quality source of active ingredients for subsequent drug development, and is both environmentally friendly and practical.

[0032] (3) This invention is the first to discover that goat milk fruit polysaccharide has the biological activity of protecting the intestinal barrier, which breaks through the traditional medicinal scope of goat milk fruit and expands its application to the treatment of intestinal barrier damage-related diseases, enriches the library of active ingredients of natural drugs, and provides new options for the treatment of related clinical diseases. Attached Figure Description

[0033] Figure 1 For glucose standard curve; Figure 2 The graph shows the effect of DES type on the extraction rate of goat milk fruit polysaccharide. Figure 3 Figure 1 shows the effect of ultrasound time on the extraction rate of polysaccharides from goat milk fruit. Figure 4 Figure 1 shows the effect of ultrasonic temperature on the extraction rate of polysaccharides from goat milk fruit. Figure 5 Figure 1 shows the effect of the volume ratio of the upper and lower phases on the extraction rate of polysaccharides from *Aegilops spp.* Figure 6 The effect of water content of DES-1 (lauric acid: decanoic acid) on the extraction rate of goat milk fruit polysaccharide is shown in the figure. Figure 7 The graph shows the effect of ammonium sulfate mass fraction on the extraction rate of goat milk fruit polysaccharides. Figure 8 shows the response surface and contour plots. In the figure, (A and D) show the effect of the interaction between ammonium sulfate mass fraction and DES moisture content on ECFP extraction rate, (B and E) show the effect of ammonium sulfate mass fraction and upper and lower phase volume ratio on extraction rate, and (C and F) show the effect of DES moisture content and upper and lower phase volume ratio on ECFP extraction rate. Figure 9 shows the molecular weight distribution of goat milk fruit polysaccharide; Figure 10 The diagram shows the monosaccharide composition of *Fructus spp.* (also known as *Fructus spp.*) polysaccharide. In the diagram, 1 represents mannose; 2 represents rhamnose; 3 represents glucuronic acid; 4 represents galacturonic acid; 5 represents glucose; 6 represents galactose; 7 represents xylose; 8 represents arabinose; and 9 represents fucose. Figure 11 The results of the effect of goat milk fruit polysaccharide on body weight; Figure 12 The results of the disease score for goat milk fruit polysaccharide; Figure 13 Results regarding the effect of goat milk fructose on colon length; Figure 14 The results of the effect of goat milk fruit polysaccharide on spleen coefficient; Figure 15 Results of H&E staining of mouse colon tissue; Figure 16 The results are from the detection of inflammatory factors in mouse colon tissue; Figure 17 The results are from the detection of oxidative stress in mouse colon tissue; Figure 18 Results of AB-PAS staining of mouse colon tissue; Figure 19 Quantitative results of immunohistochemical staining of MUC2 in mouse colon tissue; Figure 20 Quantitative results of immunohistochemical staining for ZO-1 in mouse colon tissue; Figure 21 Quantitative results of immunohistochemical staining for Occludin in mouse colon tissue; Figure 22 To demonstrate the effect of goat milk fruit polysaccharide prepared by traditional methods on the survival rate of fruit flies; Figure 23 To demonstrate the effect of goat milk fruit polysaccharide prepared by traditional methods on the intestinal permeability of Drosophila and to present a bar chart; Figure 24 To demonstrate the effect of goat milk fructose prepared by traditional methods on the intestinal morphology of Drosophila and to present a bar chart. Detailed Implementation

[0034] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0035] Furthermore, the technical solutions of the various embodiments of the present invention can be combined with each other, but only if they are feasible for those skilled in the art. If the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention.

[0036] The following specific embodiments illustrate the solution proposed in this invention: Example 1: Optimization of the extraction process of goat milk fruit polysaccharides 1. Preparation of DES Hydrogen bond acceptors (decanoic acid, lactic acid, terpineol) and hydrogen bond donors (lauric acid, betaine, glucose) were placed in 15 mL centrifuge tubes at specific molar ratios and heated in a water bath until dissolved into a clear, homogeneous liquid. DES-1: Decanoic acid + Lauric acid (1:1), DES-2: Betaine + Lactic acid (1:4), DES-3: Lauric acid + Terpineol (2:1), DES-4: Glucose + Lactic acid (1:5).

[0037] 2. Optimal DES selection Fresh *Gynura procumbens* fruit was homogenized and freeze-dried to obtain *Gynura procumbens* powder. 0.05 g of the powder was added to 1 mL of 25% ammonium sulfate solution at a 1:2 solid-liquid ratio, followed by 1 mL of DES solution in a centrifuge tube to form a three-phase distribution system. Each three-phase distribution system was replicated three times. Polysaccharides were extracted in an ultrasonic water bath at 60 ℃ for 30 min at 100 W. The lower ammonium sulfate layer was aspirated with a pipette, and the supernatant was collected after centrifugation.

[0038] 3. Calculation of the extraction rate of polysaccharides from goat milk fruit Weigh 300 mg of D-anhydrous glucose and place it in a 100 mL volumetric flask. Dissolve the glucose in a small amount of deionized water and dilute to the final volume to obtain a glucose standard stock solution. Dilute the stock solution with deionized water to obtain six groups of glucose standard solutions with concentrations of 1.5, 0.75, 0.375, 0.1875, 0.09375, and 0.046875 mg / mL. Then, pipette 30 μL of each glucose standard solution into a 96-well plate, add 30 μL of 5% α-phenol solution and 160 μL of concentrated sulfuric acid, and allow the reaction to proceed for 15 min. Measure the absorbance at 490 nm using a microplate reader. Perform the measurements three times in parallel to obtain the glucose standard curve.

[0039] The content of polysaccharides in goat milk fruit was determined according to the polysaccharide determination method in the glucose standard curve, and the calculation formula is as follows: In the formula: W: polysaccharide extraction rate (%); C: polysaccharide concentration (mg / mL); V: volume of polysaccharide solution (mL); N: dilution factor; M: mass of *Gnaphalium affine* medicinal material (g).

[0040] 4. Single-factor analysis Single-factor experiments were conducted using the polysaccharide content of *Gnaphalium affine* as the evaluation index to investigate the effects of DES type (DES-1, DES-2, DES-3, DES-4), ultrasonic extraction temperature (40℃~80℃), DES water content (15%~35%), ammonium sulfate mass fraction (15%~30%), ultrasonic extraction time (10 min~50 min), and the ratio of upper and lower phases (DES:ammonium sulfate = 0.5:1~2.5:1) on polysaccharide extraction.

[0041] 5. Response surface optimization experiment Based on the results of the single-factor experiments, three independent variables were selected: ammonium sulfate mass fraction, DES moisture content, and the ratio of upper and lower phases. The Box-Behnken response surface methodology software was used to conduct response surface analysis with the extraction rate of goat milk fruit polysaccharide as the response value to determine the optimal extraction conditions for goat milk fruit polysaccharide.

[0042] 6. Determination of molecular weight The molecular weight of polysaccharides was analyzed using high-performance liquid chromatography-gel chromatography (HPGPC). Dextran series dextran standard solutions (Dextran T70, T20, T10, T5, T2, T1) at a concentration of 4 mg / mL were prepared, filtered through a 0.22 μm microporous membrane, and then analyzed. The detection conditions were: HPLC system equipped with a TSKgel G3000PWXL gel column, mobile phase of 0.02 M KH₂PO₄ solution, and flow rate of 0.5 mL / min. A molecular weight standard curve was plotted with dextran retention time on the x-axis and the logarithm of dextran molecular weight on the y-axis. A 5 mg / mL aqueous solution of *Gnaphalium affine* polysaccharide was prepared, filtered through a 0.22 μm microporous membrane, and injected. The molecular weight of *Gnaphalium affine* polysaccharide was calculated based on the standard curve.

[0043] 7. Determination of monosaccharide composition Take 5 mg of goat milk fruit polysaccharide, treat the sample with strong acid hydrolysis, perform PMP derivatization reaction on the sample and monosaccharide standard, filter the aqueous layer through a 0.45 μm aqueous microporous membrane, and then perform HPLC analysis.

[0044] 8. Experimental Results and Analysis 8.1 Glucose Standard Curve The glucose standard curve is shown below. Figure 1 .

[0045] The results showed that within the range of 0–1.5 mg / mL, the glucose solution concentration and absorbance had a good linear relationship (R0). 2 =0.992), the regression equation of the curve is: A = 2.52617C + 0.14282, which can be used to calculate the content of polysaccharides.

[0046] 8.2 Optimal DES Selection The effect of DES type on the extraction rate of goat milk fructose is shown in [reference needed]. Figure 2 .

[0047] Experiments revealed that the highest content of goat milk fruit polysaccharides was obtained when the DES system had a lauric acid:decanoic acid ratio of 1:1. Therefore, the DES-1 system with a lauric acid to decanoic acid molar ratio of 1:1 was determined to be the optimal DES system for extracting goat milk fruit polysaccharides.

[0048] 8.3 Effect of ultrasound time The effect of ultrasound time on the extraction rate of goat milk fruit polysaccharides is shown in the figure. Figure 3 .

[0049] Experimental results showed that the polysaccharide content of *Gnaphalium affine* increased sharply from 40 to 60 min, and gradually decreased from 60 to 80 min. This is mainly because the polysaccharides were not completely dissolved in the solvent before 60 min. However, when the extraction time exceeded 60 min, some polysaccharides were degraded due to excessive sonication, resulting in a decrease in polysaccharide content. Furthermore, the longer the extraction time, the higher the experimental cost. To control costs and reduce consumption, the sonication time should be minimized. In conclusion, the optimal sonication time for *Gnaphalium affine* polysaccharides is 60 min.

[0050] 8.4 Effect of Ultrasonic Temperature The effect of ultrasonic temperature on the extraction rate of goat milk fruit polysaccharides is shown in the figure. Figure 4 .

[0051] Experimental results showed that the extraction rate of ECFP increased with increasing temperature from 40℃ to 60℃; however, when the ultrasonic temperature varied within the range of 60℃ to 80℃, the extraction rate did not show a significant change (P>0.05). Higher temperatures accelerate molecular motion in the three-phase partitioning system. As the temperature increases, more hydroxyl groups are exposed, promoting hydrogen bond formation and enhancing the hydrophilicity of the polysaccharide. This facilitates the entry of ECFP into the lower aqueous phase, thereby increasing the extraction rate. However, when the temperature exceeds a certain value, high temperatures can damage the polysaccharide structure. Furthermore, temperatures exceeding 60℃ lead to higher energy consumption. To reduce experimental costs and energy consumption, a lower extraction temperature should be used. Therefore, the optimal ultrasonic temperature for ECFP extraction is 60℃.

[0052] 8.5 The Influence of Upper and Lower Phase Proportions The effect of the ratio of upper and lower phases on the extraction rate of goat milk fruit polysaccharides is shown in the figure. Figure 5 .

[0053] Experimental results showed that the polysaccharide content of *Gnaphalium affine* increased from a phase ratio of 0.5:1 to 2:1, and decreased from 2:1 to 2.5:1. This phenomenon is mainly due to the fact that the leaching rate of polysaccharides gradually increases with the increase of solute volume. When the extraction solvent reaches a certain volume, the polysaccharides become saturated, and the leaching rate decreases. Furthermore, excessive solvent leads to waste of experimental reagents. In conclusion, the optimal phase ratio for *Gnaphalium affine* is 2:1.

[0054] 8.6 Effect of DES Moisture Content The effect of DES on the extraction rate of goat milk fructose is shown in [reference]. Figure 6 .

[0055] Experimental results showed that the polysaccharide content gradually increased when the water content of the DES-1 solvent was between 15% and 30%, reaching a maximum at 30%, and then gradually decreased. The main reason is likely that when the water content is too low, the solution system is too viscous, making polysaccharide precipitation more difficult; while as the water content increases, the solution polarity increases accordingly; when the water content exceeds a certain value, the solution polarity becomes too high, reducing the interaction force between the DES solvent and the polysaccharide, thus decreasing the polysaccharide content. In conclusion, the optimal DES water content for extracting *Gnaphalium affine* polysaccharides is 30%.

[0056] 8.7 Effect of ammonium sulfate mass fraction The effect of ammonium sulfate mass fraction on the extraction rate of goat milk fruit polysaccharides is shown in the figure. Figure 7 .

[0057] Experimental results showed that the polysaccharide content gradually increased when the ammonium sulfate mass fraction was 15% to 20%, reaching a maximum at 20%, and then gradually decreased between 20% and 30%. The main reason for this was NH4+. + and SO4 2- As the mass fraction of ammonium sulfate increases, its effect on stabilizing macromolecules also gradually increases, making the extraction system more stable. However, when the mass fraction increases to a certain value, strong salting-out occurs, reducing free water and consequently the amount of free water available to dissolve the polysaccharides, thus decreasing the polysaccharide content. In conclusion, the optimal ammonium sulfate mass fraction for extracting *Gnaphalium affine* polysaccharides is 20%.

[0058] 8.8 Results of Response Surface Experiment The results of the response surface methodology experiment are shown below. Figure 8 .

[0059] Using ECFP yield as the response value, and ammonium sulfate mass fraction (A), DES moisture content (B), and upper and lower phase ratio (C) as factors, Design-Expert software was used to draw three-dimensional response surface plots and two-dimensional response contour plots to analyze the interaction between the two independent variables in order to optimize the experimental process.

[0060] Using ammonium sulfate mass fraction (A), DES moisture content (B), and the ratio of upper and lower phases (C) as independent variables, and goat milk fruit polysaccharide content (Y) as the response value, Design-Expert software was used to plot three-dimensional response surface plots and two-dimensional response contour plots to analyze the interaction between the two independent variables and optimize the experimental process. Figure 8 As shown in Figures A and D, the contour lines are nearly circular, indicating a weak interaction between the ammonium sulfate mass fraction and the DES water content (AB); Figure 8 As shown in Figures B and E, the contour lines are elliptical, indicating a significant interaction between the ammonium sulfate mass fraction and the upper / lower phase volume ratio (AC), which greatly affects the extraction rate; Figure 8 As shown in C and F, the contour lines are elliptical, indicating a significant interaction between the moisture content of DES and the volume ratio of the upper and lower phases (BC). Based on the analysis, the optimal conditions for ultrasound-assisted extraction of *Gnaphalium affine* were determined to be: ammonium sulfate mass fraction 20.26%, DES moisture content 29.40%, and upper and lower phase ratio 1.94:1, with a theoretical maximum polysaccharide content of 56.94%. Validation experiments were conducted under these optimal conditions: ammonium sulfate mass fraction 20%, DES moisture content 30%, and upper and lower phase volume ratio 2:1. The experiment was repeated three times, and the average extraction rate of *Gnaphalium affine* polysaccharides was 54%. This indicates a high degree of agreement between the results and the predicted extraction rate of *Gnaphalium affine* polysaccharides, demonstrating that the regression model is satisfactory and highly suitable for predicting the extraction of *Gnaphalium affine* polysaccharides.

[0061] 8.9 Molecular weight of goat milk fructose The molecular weight distribution characteristics of goat milk fruit polysaccharides are shown in [reference needed]. Figure 9 .

[0062] The polysaccharide from *Gnaphalium affine* exhibits two main peaks with retention times of 11.81 min and 16.35 min. Substituting these retention times into the equation of the molecular weight standard curve, y = 14.83865 - 1.30621x + 0.06122x, further supports this finding. 2 -0.00135x 3 , R 2 =0.9994, the result shows that the molecular weight range of goat milk fruit polysaccharide is 8.53-533.70 kDa.

[0063] 8.10 Monosaccharide composition of goat milk fruit polysaccharide HPLC chromatograms of PMP pre-column derivatization of goat milk fruit polysaccharide and PMP pre-column derivatization of standard monosaccharide are shown below. Figure 10 .

[0064] Depend on Figure 10It is known that the polysaccharide of goat milk fruit is mainly composed of four monosaccharides, including galacturonic acid, glucose, galactose, and arabinose. According to the molar ratio, the content of each monosaccharide is the highest, reaching 38.77%, followed by glucose (18.43%), galactose (16.52%), and arabinose (12.71%).

[0065] Example 2: Experiment on the protective effect of goat milk fructose on the intestinal barrier 1. Grouping and processing of laboratory animals Experimental animals: 42 male SPF-grade C57BL / 6 mice, 8 weeks old, weighing 22±2g, provided by Guangdong Provincial Experimental Animal Center; Experimental grouping: The subjects were randomly divided into 6 groups, including normal group (NC group), model group (DSS group), positive drug mesalazine (5-ASA) group (150mg / kg / d), and goat milk fruit polysaccharide (ECFP) group (200mg / kg / d).

[0066] 2. Preparation of a mouse model of intestinal injury Mice were acclimatized for one week. Three days after preventative administration, a 2% sodium dextran sulfate (DSS) model was induced in the mice, with the drug administered via gavage during the modeling process. Normal mice were given distilled water to drink. The intestinal injury model was induced in mice by drinking the 2% sodium dextran sulfate solution for seven days. Simultaneously, the goat milk fructose group and mesalazine group were administered the corresponding doses of the drug via gavage, while the normal and model groups were administered an equal volume of physiological saline via gavage.

[0067] 3. The preventive and therapeutic effects of goat milk fructose on sodium dextran sulfate-induced intestinal injury in mice. Throughout the experiment, the mice's weight, defecation, and rectal bleeding were observed and recorded daily. Fecal samples were taken from each group of mice for fecal occult blood testing. Disease activity scores were calculated according to Table 1. On day 8, blood was collected via the orbital sinus, and the mice were then euthanized. The length of the colon from the proximal rectum to the ileocecal junction was measured, and the distal colon tissue was immediately fixed with 4% paraformaldehyde solution or preserved at -80°C for further analysis. The spleen was dissected and weighed using the same method.

[0068] The formula for calculating the spleen index is as follows: Histopathological analysis of colon tissue was performed. A portion of the colon was fixed in 4% paraformaldehyde, embedded in paraffin, sectioned, and stained with hematoxylin and eosin to assess the severity of intestinal damage.

[0069] Table 1 Disease Activity Scoring Criteria

[0070] 4. Experimental Results and Analysis 4.1 Effects of goat milk polysaccharide on body weight changes like Figure 11 As shown, the weight change rate in the NC group gradually increased with the number of days, showing a clear upward trend. In contrast, the weight change rate in the DSS group gradually decreased with the number of days, showing an overall downward trend, indicating that the UC mouse model was successfully established. Although the 5-ASA group showed weight loss on days 6 and 7, the decrease was significantly smaller than that in the DSS group, and the weight change rate resumed its upward trend on day 8, indicating that 5-ASA played a therapeutic role in the UC mouse model. Although the weight change rate in the goat milk fructose group (ECFP) showed a downward trend, the decrease was significantly smaller than that in the DSS group, and the downward trend on days 5 and 6 was more gradual than that in the 5-ASA group, indicating that goat milk fructose has a therapeutic effect on ulcerative colitis in mice. The above results demonstrate that the goat milk fructose of the present invention has a good protective effect against weight loss and colonic atrophy symptoms caused by intestinal damage.

[0071] 4.2 Effect of goat milk fructose on disease activity score DAI scores were negatively correlated with the severity of colitis lesions in mice; higher scores indicated more severe conditions. Figure 12 As shown, the DAI index in the NC group remained largely unchanged, consistently hovering near 0. The DAI index in the DSS group, however, showed a continuous upward trend, indicating successful UC mouse modeling. The DAI indices in both the 5-ASA and goat milk fruit polysaccharide groups fluctuated around 1, showing an overall downward trend. However, the DAI index in the goat milk fruit polysaccharide group was slightly lower than that in the 5-ASA group, indicating that the therapeutic effect of goat milk fruit polysaccharide was slightly better than that of 5-ASA, and that the mice's weight loss, fecal formation, and degree of fecal occult blood were all treated.

[0072] 4.3 Effect of goat milk fructose on colon length Ulcerative colitis in mice alters the length and shape of their intestines, such as... Figure 13 As shown, compared to the NC group, the DSS group not only had a shorter intestinal length but also exhibited multiple ulcers and bleeding points. The intestinal wall also thickened and became more fragile and prone to breakage, indicating successful UC mouse model establishment. The intestinal length of the 5-ASA group was between that of the NC and DSS groups, with a reduced number of ulcers and bleeding points, and a lessened intestinal wall thickening. The intestinal length of the goat milk fructose group was also between that of the NC and DSS groups, similar to the 5-ASA group, with almost no ulcers or bleeding points and no intestinal wall thickening. This indicates that goat milk fructose has a significant therapeutic effect on ulcerative colitis in mice, and the effect is similar to that of 5-ASA.

[0073] 4.4 Effect of goat milk fruit polysaccharide on spleen coefficient like Figure 14As shown, the spleen coefficient in the normal group was at a normal physiological level. Compared with the normal group, the spleen coefficient in the model group was significantly increased, indicating that the UC mouse model was successfully established. The spleen coefficients in both the goat milk fruit polysaccharide group and the positive control group were significantly decreased compared with the model group, and the goat milk fruit polysaccharide and the positive control group showed comparable effects. The results indicate that goat milk fruit polysaccharide can significantly reduce the DSS-induced increase in spleen coefficient and effectively alleviate systemic inflammation in UC mice.

[0074] 4.5 Pathological histological observation of goat milk fruit polysaccharide like Figure 15 As shown, H&E staining sections of NC group mice revealed relatively intact colonic crypt structures, a complete intestinal mucosal surface, clear and distinct structures, and plump goblet cells. In the DSS group, crypt structures were deformed, atrophied, or even disappeared; the overall intestinal mucosal layer was thinner, with some areas even showing complete disappearance of the mucosal layer; goblet cells atrophied, indicating successful UC mouse model establishment. Both the 5-ASA group and the goat milk fruit polysaccharide group demonstrated repair functions on the intestinal mucosa, with the intestinal crypt structure essentially restored to normal and goblet cells plump. The goat milk fruit polysaccharide group showed more complete structural repair compared to the 5-ASA group, demonstrating superior therapeutic efficacy against ulcerative colitis in mice. These results indicate that goat milk fruit polysaccharide can enhance intestinal mucosal barrier function, inhibit colonic mucosal edema, and inhibit the occurrence and development of intestinal damage induced by sodium dextran sulfate.

[0075] 4.6 Effects of goat milk fructose on serum inflammatory factors like Figure 16 As shown, compared with the NC group, the DSS group showed significantly increased levels of inflammatory factors TNF-α and IL-1β (P < 0.001, P < 0.0001), while the anti-inflammatory factor IL-10 was significantly decreased (P < 0.0001), indicating successful establishment of the UC mouse model. The levels of inflammatory factors TNF-α, IL-1β, and IL-10 in the goat milk fruit polysaccharide group were significantly lower than those in the modeling group (P < 0.05, P < 0.001), while the level of the anti-inflammatory factor IL-10 was significantly increased (P < 0.001). Furthermore, the levels of inflammatory factors in the goat milk fruit polysaccharide group were essentially the same as those in the 5-ASA group. Therefore, goat milk fruit polysaccharide has a therapeutic effect on colonic lesions.

[0076] 4.7 Effects of goat milk fructose on colonic oxidative stress like Figure 17 As shown, compared with the NC group, the MPO level in the DSS group was significantly increased (P < 0.001), indicating that the UC mouse model was successfully established. The MPO level in the goat milk fruit polysaccharide group was significantly decreased compared with the DSS group (P < 0.01), and was basically the same as that in the 5-ASA group.

[0077] The SOD level in the DSS group was significantly lower than that in the NC group (P < 0.001), indicating that the UC mouse model was successfully established. The SOD level in the goat milk fruit polysaccharide group was significantly higher than that in the DSS group (P < 0.001), and was basically the same as that in the 5-ASA group.

[0078] The GSH content in the DSS group was significantly lower than that in the NC group (P < 0.001), indicating that the UC mouse model was successfully established. The GSH content in the goat milk fruit polysaccharide group was significantly higher than that in the DSS group (P < 0.05), and its content was close to that in the NC group. At the same time, the GSH content in the 5-ASA group was also similar to that in the goat milk fruit polysaccharide group.

[0079] The MDA content in the DSS group was significantly higher than that in the NC group (P < 0.0001), indicating that the UC mouse model was successfully established. The content of goat milk fruit polysaccharide in the DSS group was significantly lower than that in the DSS group (P < 0.001), and similar to that in the NC group, while being basically the same as that in the 5-ASA group.

[0080] In conclusion, goat milk fruit polysaccharide can increase the content and activity of SOD and GSH in the colon of UC mice, while simultaneously reducing the content of MPO and MDA, indicating that goat milk fruit polysaccharide has a significant therapeutic effect on colitis in mice.

[0081] 4.8 Effects of goat milk fructose on intestinal mucoprotein like Figure 18 As shown, the NC group exhibited intact intestinal structure, abundant goblet cells, and sufficient mucus secretion, with no obvious inflammatory infiltration. Compared to the NC group, the DSS group showed a significant reduction in both the number of goblet cells and mucin, indicating that DSS-induced inflammatory responses may disrupt the integrity of the intestinal mucus barrier by consuming acidic mucin in goblet cells. The goat milk fructose administration group showed an increase in goblet cell number and restored mucus secretion, suggesting that goat milk fructose can restore goblet cell function and alleviate DSS-induced acidic mucin consumption, indicating its protective effect on the intestinal mucus barrier. Figure 19 As shown, compared with the control group, the level of mucin (MUC2) secreted by intestinal goblet cells in the DSS group mice was significantly reduced, while treatment with goat milk fruit polysaccharide significantly upregulated the protein expression of MUC2. In summary, goat milk fruit polysaccharide exerts a therapeutic effect in the UC mouse model by promoting the secretion of acidic mucin and MUC2 by goblet cells and maintaining the integrity of the intestinal mucus barrier.

[0082] 4.9 Effects of goat milk fructose on the intestinal mucosal barrier Tight junction proteins (TJs) are an important component of the intestinal mucosal barrier, effectively preventing harmful substances from leaking out of the intestinal lumen. For example... Figure 20 and 21As shown, the expression of ZO-1 and Occludin was significantly reduced in the DSS-induced UC mouse model, further confirming the disruption of intestinal barrier function. Treatment with *Elephantopus scabra* polysaccharide significantly upregulated the expression of these TJ proteins, thereby repairing the damaged intestinal barrier. These results indicate that *Elephantopus scabra* polysaccharide can significantly increase the expression level of TJ proteins, suggesting that it exerts its therapeutic effect on ulcerative colitis by repairing the integrity of the intestinal barrier.

[0083] Example 3: Experiment on the protective effect of goat milk fructose prepared by the traditional water extraction and alcohol precipitation method on the intestinal barrier. 1. Preparation of goat milk fructose polysaccharides The traditional water extraction and alcohol precipitation method was used to obtain *Cynanchum paniculatum* polysaccharide, which involves extraction with hot water followed by alcohol precipitation. 0.5 kg of dried *Cynanchum paniculatum* powder was added to 2.5 L of water at a material-to-liquid ratio of 1:5, and extracted three times at 80℃. The extracts were combined. The extracts were concentrated under reduced pressure to 1 / 10 of their original volume, and anhydrous ethanol was slowly added until the concentration reached 80%. The mixture was allowed to stand at 4℃ for 24 h, and the precipitate was collected. The precipitate was then dissolved again in an appropriate amount of deionized water, and freshly prepared Sevag reagent (dichloromethane: n-butanol = 4:1, volume ratio) was added. The mixture was vigorously shaken in a separatory funnel, allowed to stand for separation, and the supernatant was collected. This process was repeated three times until there was virtually no flocculent matter at the interface. The supernatant was placed in a 1000 Da dialysis bag and dialyzed with running water for 48 h. The dialysate was concentrated under reduced pressure and freeze-dried (48 h) to obtain crude *Cynanchum paniculatum* polysaccharide (ECFP-W).

[0084] 2. Establishment of a Drosophila model of intestinal injury Drosophila that had emerged 3-5 days prior were randomly collected and divided into a control group (5% sucrose), a model group (5% sucrose + 3.5% DSS), and a treatment group (containing 5% sucrose + 3.5% DSS). Each group had 3-4 parallel tubes, with 30 rosophila per tube. The rosophila were first starved in empty tubes for 2 hours, then transferred to tubes with three circular filter papers at the bottom. 200 μL of the treatment solution was added to each filter paper to thoroughly wet it, and the filter papers were replaced every 24 hours.

[0085] 3. Fruit fly survival rate experiment Wild-type fruit flies 2-3 days after emergence were captured and divided into 5 experimental groups: normal group (NC), model group (DSS), and three treatment groups (ECFP-W) with 1%, 2%, and 5% concentrations of fructooligosaccharide. Each group had 4 parallel tubes, with 30 fruit flies per tube. After starvation for 2 hours, the flies were transferred to tubes with 3 circular filter papers at the bottom. 200 μL of 5% sucrose solution, 3.5% DSS solution (containing 5% sucrose), and the three treatment groups (1%, 2%, and 5% fructooligosaccharide, containing 5% sucrose + 3.5% DSS) were added to each tube, respectively. The filter papers were changed every 24 hours. The number of fruit flies that died each day was recorded for 7 days. The concentration of the treatment group with the highest survival rate was obtained, and this concentration was used in subsequent experiments.

[0086] 4. Detection of intestinal barrier integrity in Drosophila (Smurf test) Wild-type female fruit flies that had emerged 2-3 days prior were randomly divided into three groups: normal group (NC), model group (DSS), and goat milk fruit polysaccharide-treated group (BCFP-W). Each group had three parallel tubes. 200 μL of brilliant blue mixed solution (i.e., the solution contained 4% brilliant blue) was added to each tube. After 4 days of culture, the fruit flies were anesthetized with isoflurane, and the surface of the fruit flies was washed with PBS to remove the surface stain. Frontal and side photographs were taken under a microscope, and the number of fruit flies stained with blue was recorded.

[0087] 5. Changes in the intestinal morphology of fruit flies Wild-type female Drosophila melanogaster (W1118) 2-3 days after emergence were captured and divided into three experimental groups: NC, DSS, and BCFP-W. After starvation for 2 hours, the flies were transferred to Drosophila tubes with three circular filter papers at the bottom. 200 μL of 5% sucrose solution, 5% sucrose solution + 3.5% DSS solution, and 5% sucrose solution + 3.5% DSS solution + 2% BCFP-W solution were added to each tube, respectively. The filter papers were changed every 24 hours for a total of 4 days. Ten to fifteen Drosophila intestines were dissected under a microscope and placed on slides containing 70% glycerol solution. The length of the intestines was observed under a microscope. Each experiment was repeated three times.

[0088] 6. Results and Analysis 6.1 Effects of goat milk polysaccharide on the survival rate of fruit flies The effect of goat milk fruit polysaccharide on the survival rate of fruit flies is shown in Figure 22 .

[0089] Depend on Figure 22It was found that the survival rate of Drosophila in the NC group remained at a high level, while the survival rate of DSS group dropped below 50% on day 5 and below 20% on day 7, indicating that DSS can significantly induce intestinal damage and death in Drosophila, successfully constructing a Drosophila lethal model related to UC. On day 7, the survival rate of Drosophila in the 1% goat milk fructooligosaccharide group was slightly higher than that in the DSS group; the survival rate of Drosophila in the 2% goat milk fructooligosaccharide group was 52%, significantly higher than that in the DSS group, with a clear protective effect; the survival rate of Drosophila in the 5% goat milk fructooligosaccharide group was also significantly higher than that in the DSS group, but the overall protective effect was weaker than that of 2% (approximately 49% on day 7), suggesting that 2% is the optimal effective concentration of goat milk fructooligosaccharide, therefore, 2% goat milk fructooligosaccharide was used in subsequent experiments.

[0090] 6.2 Effects of goat milk fructose on intestinal permeability in fruit flies The effect of goat milk fructose on intestinal permeability in fruit flies is shown in [reference needed]. Figure 23 .

[0091] Depend on Figure 23 It was observed that the normal control group (NC) showed only a small amount of blue staining in the abdomen, with no overall blue staining, indicating an intact intestinal barrier and almost no leakage. Compared to the NC group, the DSS group showed significant overall blue staining, and the Smurf positivity rate was significantly higher than that of the NC group, indicating that DSS successfully induced intestinal barrier damage in Drosophila. After the addition of goat milk fructose, the Drosophila in the treatment group showed only localized blue staining, and the positivity rate was significantly lower than that in the DSS group. The results indicate that goat milk fructose can significantly improve DSS-induced intestinal barrier damage in Drosophila, reduce the incidence of intestinal leakage, and has clear intestinal barrier protective activity.

[0092] 6.3 Effects of goat milk fructose on intestinal morphology in fruit flies The effects of goat milk fructose on the intestinal morphology of fruit flies are shown in [reference needed]. Figure 24 .

[0093] Depend on Figure 24 It can be seen that the Drosophila in the NC group had the longest intestine, while the intestine in the DSS group was significantly shorter, with a length significantly shorter than that in the NC group. This indicates that DSS successfully induced intestinal atrophy in Drosophila, and the model was successfully established. However, the intestinal length of Drosophila in the goat milk fructose-treated group was significantly increased compared to the DSS group. ** P<0.01). The results showed that goat milk fructose could significantly improve DSS-induced intestinal atrophy in Drosophila, restore intestinal length, and had clear intestinal protective and repair activities.

[0094] The difference between this invention and the prior art lies in: (1) Innovation of the research object: The research object of this invention is goat milk fruit polysaccharide. At present, no scholars have studied the intestinal protective effect of goat milk fruit polysaccharide, while this invention is the first to discover that goat milk fruit polysaccharide has the function of protecting the intestinal barrier, which can greatly expand the scope of use of goat milk fruit.

[0095] (2) Innovation in Research Methods: Currently, the extraction of plant polysaccharides both domestically and internationally mainly employs methods such as water extraction and alcohol precipitation, ultrasound-assisted extraction, and microwave-assisted extraction. The three-phase partition method is a widely used method for extracting multiple components such as oils and polysaccharides, offering many advantages but also some drawbacks. The main drawback is that the three-phase partition system typically uses volatile, flammable, and toxic tert-butanol. The goat's milk fruit polysaccharide prepared in this invention is obtained using a three-phase partition method based on a eutectic solvent, a method different from traditional methods. This invention utilizes a eutectic solvent instead of tert-butanol, solving problems such as contamination and toxicity in the original system. Furthermore, the extraction rate of goat's milk fruit polysaccharide obtained after the process optimization method of this invention reaches as high as 54%.

[0096] (3) Innovation in the research field: This invention evaluates the protective effect of goat milk fruit polysaccharide on the intestinal barrier by establishing a mouse model and a fruit fly model of intestinal barrier damage induced by dextran sulfate sodium. This efficacy is the first study on goat milk fruit polysaccharide, which is different from other literature. It also provides a new treatment method for the medical field to treat intestinal barrier-related diseases.

[0097] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0098] The sequence numbers of the above embodiments of the present invention are for descriptive purposes only and do not represent the superiority or inferiority of the embodiments.

[0099] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. Application of goat milk fruit polysaccharide in the preparation of drugs for treating intestinal barrier-related diseases.

2. The application according to claim 1, characterized in that, The intestinal barrier-related diseases include intestinal damage.

3. The application according to claim 2, characterized in that, Symptoms of intestinal damage include at least one of the following: weight loss, loose stools, fecal occult blood, shortened colon, goblet cell necrosis of colonic tissue, and mucosal edema and sloughing.

4. The method for preparing the goat milk fruit polysaccharide according to any one of claims 1 to 3, characterized in that, The process involves drying the powdered goat's milk fruit, extracting the powdered goat's milk fruit in a three-phase distribution system containing an organic solvent and a salting-out solution under a set extraction temperature and ultrasonic assistance. After extraction, the aqueous phase is concentrated and dried to obtain goat's milk fruit polysaccharide. The organic solvent used is a eutectic solvent, and the salting-out solution is an aqueous solution of ammonium sulfate.

5. The method according to claim 4, characterized in that, The eutectic solvent is composed of a hydrogen bond acceptor and a hydrogen bond donor reagent. The hydrogen bond acceptor is selected from any one of decanoic acid, terpineol, and lactic acid, and the hydrogen bond donor is selected from any one of lauric acid, betaine, and glucose.

6. The method according to claim 5, characterized in that, The molar ratio of the hydrogen bond donor to the hydrogen bond acceptor is 1:1 to 2:

1.

7. The method according to claim 5, characterized in that, The hydrogen bond donor is lauric acid, the hydrogen bond acceptor is decanoic acid, and the molar ratio of lauric acid to decanoic acid is 1:

1.

8. The method according to claim 4, characterized in that, The mass fraction of ammonium sulfate in the aqueous solution used is 15-30%.

9. The method according to claim 4, characterized in that, The organic solvent has a water content of 15-35%; the volume ratio of the organic solvent to the salting-out solution is 0.5:1 to 2.5:

1. The extraction temperature is 40℃~80℃, and the ultrasonic extraction time is 40min~80min.

10. The goat milk fruit polysaccharide prepared by the method according to any one of claims 4-9, characterized in that, The monosaccharide composition of the goat milk fruit polysaccharide, in molar ratio, includes 35-40% galacturonic acid, 15-20% glucose, 15-20% galactose, and 10-15% arabinose; the molecular weight of the goat milk fruit polysaccharide is 8.0-535 kDa.

Images