Small molecular weight Fritillaria thunbergii polysaccharide, Fritillaria thunbergii polysaccharide nano-selenium complex, preparation method thereof and application

Abstract

The present invention provides a small-molecular-weight Fritillaria thunbergii Miq. polysaccharide, which comprises galactose, arabinose, galacturonic acid, glucose and rhamnose; the molecular weight of the small-molecular-weight Fritillaria thunbergii Miq. polysaccharide is 50-100 kDa. Compared with the prior art, the small-molecular-weight Fritillaria thunbergii Miq. polysaccharide provided by the present invention has a smaller molecular weight, is more easily absorbed and utilized by the body, and has a better anti-inflammatory effect than the high-molecular-weight Fritillaria thunbergii Miq. polysaccharide.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and particularly relates to small molecule fritillary bulb polysaccharide, fritillary bulb polysaccharide nano-selenium complex, its preparation method and application. Background Art

[0002] There are a large number of pathogenic bacteria in daily life. Under normal circumstances, these pathogenic bacteria will not invade the human body and cause harm to the human body. However, when the external environment changes, such as being ill, having low immunity, having a wound, being infected, etc., the pathogenic bacteria will take advantage of the weakness to invade and cause a series of inflammatory reactions in the body, prolong the disease treatment time, bring unnecessary pain to the patient, and even endanger life seriously in severe cases.

[0003] Common pathogenic bacteria in life include staphylococcus, streptococcus, escherichia coli, etc. These bacteria can cause symptoms such as wound infection and suppuration, diarrhea, etc. Strengthening protection can greatly reduce the infection probability of pathogenic bacteria and protect the health of the body.

[0004] Fritillary bulb polysaccharide is an important nutrient component in the bulbs of fritillary thunbergii. Fritillary bulb polysaccharide is mainly composed of glucose, galactose, arabinose, and galacturonic acid, and contains structural domains such as amyloid glucan, polyrhamnogalacturonan-type pectin, and arabinogalactan. Fritillary bulb polysaccharide has various effects such as enhancing immunity, anti-tumor, and inhibiting inflammation. Natural selenized polysaccharides often exist in plants, but the content is very low. Selenized polysaccharides have various biological activities such as antioxidant, antibacterial, and anti-tumor, and are easily absorbed and utilized by the body, and can be added to products as active ingredients.

[0005] Because fritillary bulb polysaccharide contains a large number of sugar residues such as glucose and galactose, the structure of the polysaccharide itself is relatively stable. The reported molecular weights of fritillary bulb polysaccharide are relatively large at present, which are not easily absorbed by the body, and the selenization reaction is not easy to carry out and the generated products are unstable, which hinders the application of selenized fritillary bulb polysaccharide. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a small molecule fritillary bulb polysaccharide, a fritillary bulb polysaccharide nano-selenium complex, its preparation method and application that are easy to absorb and have good anti-inflammatory effects.

[0007] The present invention provides a small molecular weight fritillary bulb polysaccharide, including galactose, arabinose, galacturonic acid, glucose and rhamnose; the molecular weight of the small molecular weight fritillary bulb polysaccharide is 50-100 kDa.

[0008] Preferably, the molar ratio of galactose, arabinose, galacturonic acid to glucose is (28-34):(16-22):(10-14):(32-38).

[0009] The present invention also provides a method for preparing small molecular weight fritillary bulb polysaccharide, comprising the following steps:

[0010] S1) Mix fritillary bulb, compound enzyme and water, then heat for extraction and filter to obtain a filtrate; the compound enzyme includes papain, pectinase and amylase;

[0011] S2) Concentrate the filtrate and centrifuge to obtain a supernatant;

[0012] S3) Adjust the pH value of the supernatant to weakly acidic for acid hydrolysis reaction, and then adjust the pH value to weakly alkaline for alkaline hydrolysis reaction to obtain a hydrolysis solution;

[0013] S4) Filter and dialyze the hydrolysis solution to obtain a dialysate;

[0014] S5) Concentrate the dialysate and add ethanol for precipitation to obtain small molecular weight fritillary bulb polysaccharide.

[0015] Preferably, the mass ratio of papain, pectinase and amylase is 1:(2-4):(1-3); the mass ratio of fritillary bulb to compound enzyme is (100-300):1.

[0016] Preferably, the temperature for heating extraction in step S1) is 50°C - 70°C;

[0017] In step S2), it is concentrated to 1 / 20 - 1 / 5 of the volume of the filtrate;

[0018] In step S3), the pH value for acid hydrolysis reaction is 5 - 6; the temperature for acid hydrolysis reaction is 2°C - 6°C; the time for acid hydrolysis reaction is 8 - 15 h;

[0019] The pH value for alkaline hydrolysis reaction is 7.8 - 8.5; the temperature for alkaline hydrolysis reaction is 2°C - 6°C; the time for alkaline hydrolysis reaction is 8 - 15 h;

[0020] In step S3), 0.01 - 0.5 mol / L trifluoroacetic acid aqueous solution is used to adjust the pH value of the supernatant to weakly acidic;

[0021] 0.1 - 0.3 mol / L sodium bicarbonate aqueous solution is used to adjust the pH value to weakly alkaline;

[0022] In step S4), the cut-off molecular weight of the membrane used for filtration and dialysis is 1200 - 1800 Da;

[0023] In step S5), it is concentrated to 1 / 4 - 1 / 2 of the volume of the dialysate; the amount of ethanol added makes the ethanol concentration in the system 75% - 85%.

[0024] The present invention also provides a fritillary bulb polysaccharide nano-selenium complex, which is obtained by selenizing the above-mentioned low-molecular-weight fritillary bulb polysaccharide.

[0025] The present invention also provides a preparation method of a fritillary bulb polysaccharide nano-selenium complex, comprising the following steps:

[0026] A1) Mix the low-molecular-weight fritillary bulb polysaccharide described in claim 1 or 2 or the low-molecular-weight fritillary bulb polysaccharide prepared by the preparation method described in any one of claims 3 to 5, sodium selenite and a reducing agent, and heat for a reduction reaction to obtain a nano-selenium sol;

[0027] A2) Remove the reducing agent from the nano-selenium sol by high-speed centrifugation or dialysis to obtain a fritillary bulb polysaccharide nano-selenium complex.

[0028] Preferably, the molar ratio of sodium selenite to the reducing agent is 1:(3-25); the concentration of the low-molecular-weight fritillary bulb polysaccharide in the nano-selenium sol is 300-1200 mg / L.

[0029] Preferably, the temperature of the reduction reaction is 40°C - 85°C; the time of the reduction reaction is 0.2 - 5 h;

[0030] The rotation speed of the high-speed centrifugation separation is 7500 - 12000 r / min; the time of the high-speed centrifugation is 10 - 35 min;

[0031] The cut-off molecular weight of the dialysis membrane used for dialysis is 3 - 10000 kDa; the time of dialysis is 6 - 48 h.

[0032] The present invention also provides the application of the low-molecular-weight fritillary bulb polysaccharide and / or the fritillary bulb polysaccharide nano-selenium complex in the preparation of antibacterial and anti-inflammatory drugs and / or antibacterial and anti-inflammatory daily chemical products;

[0033] The small-molecule fritillary bulb polysaccharide is the above-mentioned low-molecular-weight fritillary bulb polysaccharide;

[0034] The fritillary bulb polysaccharide nano-selenium complex is the above-mentioned fritillary bulb polysaccharide nano-selenium complex.

[0035] The present invention provides a low-molecular-weight fritillary bulb polysaccharide, which includes galactose, arabinose, galacturonic acid, glucose and rhamnose; the molecular weight of the low-molecular-weight fritillary bulb polysaccharide is 50 - 100 kDa. Compared with the prior art, the low-molecular-weight fritillary bulb polysaccharide provided by the present invention has a smaller molecular weight, is more easily absorbed and utilized by the body, and has a better anti-inflammatory effect than the high-molecular-weight fritillary bulb polysaccharide.

[0036] Furthermore, the present invention provides a preparation method of a low-molecular-weight fritillary bulb polysaccharide, which is obtained by acid-enzyme complex hydrolysis, and the preparation method is simple.

[0037] Furthermore, the present invention provides a fritillary bulb polysaccharide nano-selenium complex. Through the selenization reaction of small molecular weight fritillary bulb polysaccharide and selenium solution under specific conditions, this selenization reaction is easier to carry out and the reaction is relatively complete. The obtained product is stable and the process route is feasible.

[0038] Furthermore, the present invention confirms through in vivo and in vitro experiments that small molecular weight fritillary bulb polysaccharide and its nano-selenium complex can inhibit the proliferation of pathogenic bacteria and regulate inflammatory factors, indicating that small molecular weight fritillary bulb polysaccharide and its nano-selenium complex have the effects of antibacterial and anti-inflammatory. And this small molecular weight fritillary bulb polysaccharide and its nano-selenium complex are derived from the medicinal plant fritillary bulb, which is safe to use and has no toxic and side effects when used to inhibit pathogenic bacteria and reduce inflammation. Description of the Drawings

[0039] Figure 1 It is the infrared spectrogram of the small molecular weight fritillary bulb polysaccharide obtained in Example 1 of the present invention;

[0040] Figure 2 It is the scanning electron micrograph of the small molecular weight fritillary bulb polysaccharide obtained in Example 1 of the present invention;

[0041] Figure 3 It is the high performance liquid chromatogram of the monosaccharide composition of the small molecular weight fritillary bulb polysaccharide obtained in Example 1;

[0042] Figure 4 It is the molecular weight distribution diagram of the small molecular weight fritillary bulb polysaccharide obtained in Example 1 of the present invention;

[0043] Figure 5 It is the result diagram of the influence of the small molecular weight fritillary bulb polysaccharide obtained in Example 1 of the present invention on inflammatory factors. Detailed Embodiments

[0044] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than 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 work fall within the scope of protection of the present invention.

[0045] The present invention provides a small molecular weight fritillary bulb polysaccharide, which includes galactose, arabinose, galacturonic acid, glucose and rhamnose; the molecular weight of the small molecular weight fritillary bulb polysaccharide is 50-100 kDa.

[0046] Among them, the molar ratio of the galactose, arabinose, galacturonic acid to glucose is preferably (28 - 34):(16 - 22):(10 - 14):(32 - 38), more preferably (30 - 32):(17 - 21):(11 - 13):(34 - 36), still more preferably 31:(18 - 20):12:35, and most preferably 31:19:12:35.

[0047] The content of rhamnose in the small molecular weight fritillary bulb polysaccharide is preferably less than 3%.

[0048] In the present invention, the small molecular weight fritillary bulb polysaccharide further includes mannose and glucuronic acid; the contents of both mannose and glucuronic acid are lower than 0.5%.

[0049] The small molecular weight fritillary bulb polysaccharide provided by the present invention has a smaller molecular weight, is more easily absorbed and utilized by the body, and has a better anti-inflammatory effect than the high molecular weight fritillary bulb polysaccharide.

[0050] The present invention also provides a preparation method of the above-mentioned small molecular weight fritillary bulb polysaccharide, comprising the following steps: S1) Mix fritillary bulb, complex enzyme and water, then heat and extract, and filter to obtain a filtrate; the complex enzyme includes papain, pectinase and amylase; S2) Concentrate the filtrate and centrifuge to obtain a supernatant; S3) Adjust the pH value of the supernatant to weakly acidic for acid hydrolysis reaction, and then adjust the pH value to weakly alkaline for alkaline hydrolysis reaction to obtain a hydrolysis solution; S4) Filter and dialyze the hydrolysis solution to obtain a dialysate; S5) Concentrate the dialysate and add ethanol for precipitation to obtain the small molecular weight fritillary bulb polysaccharide.

[0051] The small molecular weight fritillary bulb polysaccharide obtained by acid-enzyme complex hydrolysis in the present invention has a simple preparation method.

[0052] Among them, the present invention does not have any special restrictions on the sources of all raw materials, and they can be commercially available.

[0053] Mix fritillary bulb, complex enzyme and water; in the present invention, it is preferred to soak the washed fritillary bulb in water and then add the complex enzyme; the ratio of fritillary bulb to water is preferably (0.1 - 1) kg:10 L, more preferably (0.3 - 0.8) kg:10 L, still more preferably 0.5 kg:10 L; the complex enzyme includes papain, pectinase and amylase; the mass ratio of papain, pectinase and amylase is preferably 1:(2 - 4):(1 - 3), more preferably 1:(2.5 - 3.5):(1.5 - 2.5), still more preferably 1:3:2; the mass ratio of fritillary bulb to complex enzyme is preferably (100 - 300):1, more preferably (150 - 250):1, still more preferably 200:1.

[0054] After mixing, perform heating extraction; the temperature for the heating extraction is preferably 50°C to 70°C, more preferably 55°C to 65°C, and even more preferably 60°C; the time for the heating extraction is preferably 20 to 80 min, more preferably 40 to 60 min, and even more preferably 50 min; in the present invention, it is preferred to perform heating extraction multiple times; the number of times of heating extraction is preferably 2 to 3 times.

[0055] After the heating extraction, filter to obtain a filtrate; the mesh size of the filter for the filtration is preferably 80 to 150 mesh, more preferably 100 mesh; the filter can be a filter well-known to those skilled in the art, and there is no special limitation. In the present invention, it is preferably gauze.

[0056] Concentrate the filtrate and centrifuge to obtain a supernatant; the concentration is preferably to 1 / 20 to 1 / 5 of the volume of the filtrate, more preferably to 1 / 20 to 1 / 10 of the volume of the filtrate; the method of concentration can be a method well-known to those skilled in the art, and there is no special limitation. In the present invention, it is preferably constant-temperature water bath concentration; the temperature for the concentration is preferably 50°C to 70°C, more preferably 60°C; the rotation speed for the centrifugation is preferably 3000 to 6000 rpm, more preferably 4000 to 5000 rpm, and even more preferably 4500 rpm; the time for the centrifugation is preferably 10 to 20 min, more preferably 15 min.

[0057] Adjust the pH value of the supernatant to weakly acidic for acid hydrolysis reaction; in the present invention, it is preferably to adjust the pH value of the supernatant to weakly acidic with a 0.01 to 0.5 mol / L trifluoroacetic acid aqueous solution; the concentration of the trifluoroacetic acid aqueous solution is preferably 0.05 to 0.3 mol / L, and even more preferably 0.1 to 0.2 mol / L; the pH value of the weakly acidic is preferably 5 to 6, more preferably 5.4 to 5.8, and even more preferably 5.6; the temperature for the acid hydrolysis reaction is preferably 2°C to 6°C, more preferably 4°C; the time for the acid hydrolysis reaction is preferably 8 to 15 h, more preferably 10 to 14 h, and even more preferably 12 h.

[0058] Then adjust the pH value to weakly basic for base hydrolysis reaction to obtain a hydrolysis solution; in the present invention, it is preferably to adjust the pH value to weakly basic with a 0.1 to 0.3 mol / L sodium bicarbonate aqueous solution; the pH value of the sodium bicarbonate aqueous solution is preferably 0.2 mol / L; the pH value of the weakly basic is preferably 7.8 to 8.5, more preferably 8 to 8.5, and even more preferably 8.2; the temperature for the base hydrolysis reaction is preferably 2°C to 6°C, more preferably 4°C; the time for the base hydrolysis reaction is preferably 8 to 15 h, more preferably 10 to 14 h, and even more preferably 12 h.

[0059] Filter and dialyze the hydrolyzed solution to obtain a dialysate; the molecular weight cut-off of the membrane used for filtration and dialysis is preferably 1200-1800 Da, more preferably 1400-1600 Da, and still more preferably 1500 Da.

[0060] Concentrate the dialysate and then add ethanol for precipitation; the concentration is preferably to 1 / 4-1 / 2 of the volume of the dialysate, more preferably 1 / 3-1 / 2; the temperature of the concentration is preferably 50°C-70°C, more preferably 55°C-65°C, and still more preferably 60°C; the amount of ethanol added makes the concentration of ethanol in the system preferably 75%-85%, more preferably 80%; the temperature of the precipitation is preferably 2°C-6°C, more preferably 4°C; the time of the precipitation is preferably 8-15 h, more preferably 10-14 h, and still more preferably 12 h.

[0061] After precipitation, preferably centrifuge, wash, and dry to obtain small molecular weight Fritillaria thunbergii Miq. polysaccharide; the rotation speed of the centrifugation is preferably 3000-6000 rpm, more preferably 4000-5000 rpm, and still more preferably 4500 rpm; the time of the centrifugation is preferably 10-20 min, more preferably 15 min; the washing is preferably carried out with ethanol; the number of washing times is preferably 2-3 times; the temperature of the drying is preferably 50°C-70°C, more preferably 55°C-65°C, and still more preferably 60°C.

[0062] The present invention also provides a Fritillaria thunbergii Miq. polysaccharide nano-selenium complex, which is obtained by selenizing the above-mentioned small molecular weight Fritillaria thunbergii Miq. polysaccharide.

[0063] The present invention also provides a preparation method of a Fritillaria thunbergii Miq. polysaccharide nano-selenium complex, comprising the following steps: A1) Mix the above-mentioned small molecular weight Fritillaria thunbergii Miq. polysaccharide, sodium selenite and a reducing agent, and heat for a reduction reaction to obtain a nano-selenium sol; A2) Remove the reducing agent from the nano-selenium sol by high-speed centrifugation separation or dialysis separation to obtain a Fritillaria thunbergii Miq. polysaccharide nano-selenium complex.

[0064] Among them, the present invention has no special restrictions on the sources of all raw materials, and they can be commercially available.

[0065] Mix small molecular weight fritillary bulb polysaccharide, sodium selenite and a reducing agent, and heat them to carry out a reduction reaction to obtain nano-selenium sol; the reducing agent can be a reducing agent well-known to those skilled in the art without special limitations, and vitamin C is preferably used in the present invention; the molar ratio of sodium selenite to the reducing agent is preferably 1:(3-25), more preferably 1:(5-25), and still more preferably 1:(15-25); the ratio of the small molecular weight fritillary bulb polysaccharide to sodium selenite is preferably (100-1000) mg:1 mmol, more preferably (300-800) mg:1 mmol, still more preferably (400-600) mg:1 mmol, and most preferably (450-500) mg:1 mmol; in the present invention, the small molecular weight fritillary bulb polysaccharide, sodium selenite and the reducing agent are preferably mixed in the form of their respective aqueous solutions; the concentration of the small molecular weight fritillary bulb polysaccharide aqueous solution is preferably 500-1000 mg / L, more preferably 600-900 mg / L, still more preferably 700-900 mg / L, and most preferably 800 mg / L; the concentration of the sodium selenite solution is preferably 1-10 mmol / L, more preferably 3-8 mmol / L, still more preferably 5-6 mmol / L; the concentration of the reducing agent solution is preferably 100-500 mmol / L, more preferably 200-400 mmol / L, still more preferably 300 mmol / L; the temperature of the reduction reaction is preferably 40°C-85°C, more preferably 50°C-70°C, still more preferably 50°C-60°C, and most preferably 55°C; the time of the reduction reaction is preferably 0.2-5 h, more preferably 1-4 h, still more preferably 2-3 h, and most preferably 2.5 h; the reduction reaction is preferably carried out under static conditions; the concentration of the small molecular weight fritillary bulb polysaccharide in the obtained nano-selenium sol is preferably 300-1200 mg / L.

[0066] Remove the reducing agent from the nano-selenium sol by high-speed centrifugation or dialysis; the rotation speed of the high-speed centrifugation is preferably 7500-12000 r / min, more preferably 8000-11000 r / min, and still more preferably 9000-10000 r / min; the time of the high-speed centrifugation is preferably 10-35 min, more preferably 10-30 min, and still more preferably 10-20 min; the temperature of the high-speed centrifugation is preferably 2°C-6°C, more preferably 4°C; the cut-off molecular weight of the dialysis membrane used for dialysis is preferably 3-10000 kDa; the time of the dialysis is preferably 6-48 h.

[0067] Finally, freeze-drying is preferably carried out to obtain fritillary bulb polysaccharide nano-selenium complex.

[0068] The present invention also provides the application of the above-mentioned small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano-selenium complex in the preparation of antibacterial and anti-inflammatory drugs and / or antibacterial and anti-inflammatory daily chemical products.

[0069] In this invention, in vivo and in vitro experiments have confirmed that small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano-selenium complex can inhibit the proliferation of pathogenic bacteria and regulate inflammatory factors, indicating that small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano-selenium complex have the effects of antibacterial and anti-inflammatory. Moreover, the small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano-selenium complex provided by this invention are derived from the medicinal plant fritillary bulb, are safe to use, and have no toxic or side effects when used to inhibit pathogenic bacteria and reduce inflammation.

[0070] In this invention, the antibacterial and anti-inflammatory drug achieves its effects by inhibiting the proliferation of bacteria, regulating the levels of inflammatory factors, and enhancing the body's immunity.

[0071] The pathogenic bacteria targeted by the antibacterial and anti-inflammatory drug are common pathogenic bacteria in daily life, such as staphylococcus, streptococcus, and escherichia coli, etc.

[0072] Preferably, the antibacterial and anti-inflammatory drug further includes pharmaceutically acceptable excipients; the excipients preferably include solvents, cosolvents, emulsifiers, colorants, binders, fillers, lubricants, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, antiadhesives, chelating agents, penetration enhancers, pH regulators, buffers, surfactants, thickeners, humectants, absorbents, diluents, flocculants and deflocculants, filter aids, etc.

[0073] The dosage form of the drug can be any dosage form well-known to those skilled in the art without special limitations. In this invention, it can be: sprays, aerosols, powder aerosols, lotions, ointments, liniments, nasal sprays, effervescent tablets, gargles, powders, emulsions, suspensions, solutions.

[0074] In a specific embodiment provided by this invention, as an effective antibacterial and anti-inflammatory agent, small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano-selenium complex can inhibit the proliferation of bacteria in the respiratory tract, reduce the levels of inflammatory factors, promote the recovery of the immunity of respiratory epithelial cells, and alleviate the inflammatory reaction caused by upper respiratory tract infection when administered by nasal drops or nasal spray or orally.

[0075] In a specific embodiment provided by the present invention, the drug for inhibiting bacteria and reducing inflammation is an inhalant for inhibiting bacteria and reducing inflammation; preferably, 35-55 grams of sodium chloride, 60-120 grams of citric acid, 60-80 grams of sodium citrate, 50-100 grams of small molecular weight fritillaria thunbergii polysaccharide and / or fritillaria thunbergii polysaccharide nano selenium complex are included in 15-25 L of the inhalant; in the present invention, preferably 1-2.5 grams of a preservative is further included; the preservative can be a preservative well-known to those skilled in the art, and there is no special limitation. In the present invention, benzalkonium chloride is preferably used; the above-mentioned inhalant for inhibiting bacteria and reducing inflammation is preferably prepared according to the following method: sodium chloride, citric acid, and sodium citrate are stirred and dissolved with 5-40 times the amount of water respectively; the small molecular weight fritillaria thunbergii polysaccharide and / or fritillaria thunbergii polysaccharide nano selenium complex are stirred and dissolved with 5-10 times the amount of water; the above solutions are combined, fixed in volume with water, filtered and sterilized to obtain the inhalant for inhibiting bacteria and reducing inflammation; the pore diameter of the filter membrane used for the filtration and sterilization is preferably 0.5 microns.

[0076] In a specific embodiment provided by the present invention, the drug for inhibiting bacteria and reducing inflammation is an oral liquid for inhibiting bacteria and reducing inflammation; the oral liquid for inhibiting bacteria and reducing inflammation preferably includes water extracts of wolfberry fruits, water extracts of lilies, small molecular weight fritillaria thunbergii polysaccharide and / or fritillaria thunbergii polysaccharide nano selenium complex, saccharide substances, citric acid and table salt; by raw material mass, 8-12 L of the oral liquid for inhibiting bacteria and reducing inflammation includes: 0.5-3 kg of wolfberry fruits, 0.3-1 kg of lilies, 150-450 grams of small molecular weight fritillaria thunbergii polysaccharide and / or fritillaria thunbergii polysaccharide nano selenium complex, 1000-2100 grams of saccharide substances, 30-60 grams of citric acid and 2-20 grams of table salt; the saccharide substances preferably include honey and white sugar; the mass ratio of the honey to the white sugar is preferably (7-11):(3-10); the water extracts of wolfberry fruits and lilies are preferably prepared according to the following method: the wolfberry fruits and lilies are boiled with water, and extraction is carried out while keeping the fire low, and after filtration, the water extracts of wolfberry fruits and lilies are obtained; the ratio of the wolfberry fruits to water is preferably (0.5-3) kg:(15-25) L; the extraction time is preferably 1-3 h; after filtration, concentration is preferably further carried out; the concentration ratio is preferably 1 / 3-2 / 3 of the volume of the filtrate; the oral liquid for inhibiting bacteria and reducing inflammation is preferably prepared according to the following method: small molecular weight fritillaria thunbergii polysaccharide and / or fritillaria thunbergii polysaccharide nano selenium complex and honey are added to the water extracts of wolfberry fruits and lilies, and after natural cooling, standing is carried out. After complete precipitation, filtration is carried out, then white sugar, citric acid and table salt are added, and after pressure filtration, filtration through a microporous membrane is carried out. Finally, the filtrate is pasteurized to obtain the oral liquid for inhibiting bacteria and reducing inflammation; the standing time is preferably 1-5 h; the filtration through the microporous membrane is preferably two-stage filtration through a microporous membrane; the pore diameter of the first-stage microporous membrane is preferably 3-8 microns, more preferably 5 microns; the pore diameter of the second-stage microporous membrane is preferably 0.5 microns; the temperature of the pasteurization is preferably 80°C-95°C; the time of the pasteurization is preferably 40-60 min.

[0077] The daily chemical products mentioned above can be those well-known to those skilled in the art, such as body lotion, hand lotion, hand sanitizer, mouthwash, disinfectant, etc.

[0078] The antibacterial and anti-inflammatory daily chemical products can be obtained by adding fritillaria thunbergii polysaccharide with an average molecular weight and / or fritillaria thunbergii polysaccharide nano-selenium complex during the preparation process of the daily chemical products.

[0079] In a specific embodiment provided by the present invention, the antibacterial and anti-inflammatory daily chemical product is preferably an antibacterial and anti-inflammatory body lotion; the antibacterial and anti-inflammatory body lotion preferably comprises an oil phase and an aqueous phase; by weight, the oil phase comprises 3-6 parts of stearic acid, 0.5-5 parts of monoglyceryl stearate, 1-4 parts of isopropyl palmitate, 1-4 parts of petrolatum, 2-8 parts of white mineral oil, 1-4 parts of cetyl alcohol, 0.1-0.6 parts of essence, 0.1-0.5 parts of methyl paraben and 0.1-0.3 parts of propyl paraben; by weight, the aqueous phase comprises 2-10 parts of glycerol, 0.1-1 part of allantoin, 0.1-0.5 part of 2,4-dichlorophenol, 0.1-0.8 part of triethanolamine, 0.2-10 parts of fritillaria thunbergii polysaccharide with a small molecular weight and / or fritillaria thunbergii polysaccharide nano-selenium complex and 50-80 parts of water; in the present invention, the antibacterial and anti-inflammatory body lotion is preferably prepared according to the following method: heating and stirring the oil phase (stearic acid, monoglyceryl stearate, isopropyl palmitate, petrolatum, white mineral oil, cetyl alcohol, methyl paraben, propyl paraben) until dissolved; dissolving the fritillaria thunbergii polysaccharide with a small molecular weight and / or fritillaria thunbergii polysaccharide nano-selenium complex in water, then adding glycerol and triethanolamine, and heating and stirring until dissolved; then slowly pouring the aqueous phase into the oil phase under vigorous stirring; after treatment with a homogenizer, start heating and stirring, add allantoin, 2,4-dichlorophenol and essence when cooled to 45°C - 55°C, and continue stirring and cooling to 25 - 35°C before discharging; let it stand for 2 - 3 days without change, and package after passing the inspection.

[0080] In a specific embodiment provided by the present invention, the daily chemical product with antibacterial and anti-inflammatory effects is preferably an antibacterial and anti-inflammatory hand sanitizer; calculated by weight, the antibacterial and anti-inflammatory hand sanitizer preferably comprises 30-85 parts of water, 1.1-1.6 parts of sodium dodecyl sulfate, 0.5-1.2 parts of sodium sulfite, 1.0-1.5 parts of sodium chloride, 2-6 parts of AES, 0.1-0.8 parts of glycerol, 1.2-4.0 parts of coconut oil fatty acid diethanolamide, and 1.0-10 parts of small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano selenium complex; in the present invention, the antibacterial and anti-inflammatory hand sanitizer is preferably prepared according to the following method: place water in a beaker, put it in a water bath pot, keep it at a constant temperature of 45°C, add sodium dodecyl sulfate, sodium sulfite, and sodium chloride, stir and dissolve, then add AES, continue to stir and dissolve, add glycerol and coconut oil fatty acid diethanolamide, after the liquid becomes viscous, take out the beaker, cool it to room temperature, stir evenly, and divide it into portions.

[0081] The small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano selenium complex provided by the present invention has antibacterial, moisturizing, and repair effects when used externally, and oral administration of fritillary bulb polysaccharide has effects such as enhancing immunity, antioxidant, antibacterial, and anti-inflammatory, and both external use and oral administration are beneficial and harmless to the human body.

[0082] The small molecular weight fritillary bulb polysaccharide and / or fritillary bulb polysaccharide nano selenium complex provided by the present invention can be prepared into an inhalant (nasal spray), enter the nasal cavity, trachea, and lungs to prevent bacterial growth and be used for antibacterial and anti-inflammatory; adding fritillary bulb polysaccharide and its nano selenium complex to the body lotion formula can increase the effect of the body lotion antibacterial and anti-inflammatory product and prevent bacterial infection; the oral liquid prepared from fritillary bulb polysaccharide and its nano selenium complex can inhibit the invasion of pathogenic bacteria from the digestive system and play a role in preventing and treating bacterial infection and eliminating inflammation.

[0083] In order to further illustrate the present invention, the following examples are used to describe in detail a small molecular weight fritillary bulb polysaccharide, fritillary bulb polysaccharide nano selenium complex, its preparation method, and application provided by the present invention.

[0084] The reagents used in the following examples are all commercially available. The CAS numbers of papain, pectinase, and amylase used in the examples are: 9001-73-4, 9032-75-1, and 9000-90-2 respectively, and they are all purchased from Shanghai Yuanye Bio-Technology Co., Ltd.

[0085] Example 1

[0086] Wash 0.5 kg of Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia, add it to 10 L of water and soak, add 2.5 g of the self-made composite enzyme in the laboratory (the mass ratio of papain, pectinase, and amylase is 1:3:2), boil and extract twice with 60 °C hot water, filter with four layers of 100-mesh gauze, concentrate the filtrate to 1 L, centrifuge (4000 rpm, 10 min, room temperature), and take the supernatant; add 0.01 mol / L trifluoroacetic acid aqueous solution to the supernatant, adjust the pH to 5.6, stir magnetically at 4 °C for 12 h to carry out acid hydrolysis reaction; add 0.2 mol / L sodium bicarbonate aqueous solution, adjust the pH to 8.2, stir magnetically at 4 °C for 12 h to carry out alkali hydrolysis; the solution after acid and alkali hydrolysis is filtered and dialyzed with a membrane (molecular weight 1500 Da), after the dialysate is concentrated to 500 mL at 60 °C, add anhydrous ethanol solution until the final ethanol concentration is 80%, place at 4 °C for 12 h, centrifuge (4000 rpm, 10 min, room temperature) to take the precipitate; wash the precipitate with anhydrous ethanol 2 to 3 times, then centrifuge (4000 rpm, 10 min, room temperature), take the precipitate, and dry at 60 °C to obtain small molecular weight Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia polysaccharide.

[0087] The small molecular weight Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia polysaccharide solution (800 mg / L) is successively mixed with sodium selenite solution (60 mM) and vitamin C (300 mM) solution, with a volume ratio of 170:5:25, and left standing in a water bath at 55 °C for 2.5 h, separated by high-speed centrifugation (10000 rpm, 10 min, room temperature) to remove vitamin C, and a nano selenium suspension is obtained. After freeze-drying (-40 °C, 36 h, vacuum), a solid powder-like Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia polysaccharide nano selenium complex is obtained.

[0088] This example may also include pretreatment steps such as solution preparation and preparation of ultrapure water. Example 2

[0089] Determine the structural characteristics and composition of the small molecule Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia polysaccharide and its nano selenium complex prepared in Example 1. The specific method is as follows:

[0090] Use infrared spectroscopy to detect the structural characteristics of the small molecule Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia polysaccharide, and the results Figure 1 are shown.

[0091] Use electron microscopy scanning to detect the spatial conformation of the small molecule Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia polysaccharide, and the results Figure 2 are shown. The monosaccharide composition of the small molecule Fritillaria thunbergii Miq. var. chekiangensis Hsiao et K. C. Hsia polysaccharide obtained in Example 1 is determined by pre-column PMP derivation-high performance liquid chromatography (PMP-HPLC), and its high performance liquid chromatography diagram is as Figure 3As shown below. The specific method is as follows: Weigh 2 mg of the polysaccharide sample and perform complete acid hydrolysis. Add 0.5 mL of 2 M hydrochloric acid methanol solution, seal the tube with N2, hydrolyze at 80 °C for 16 h. After air drying, add 0.5 mL of 2 M trifluoroacetic acid, hydrolyze at 120 °C for 1 h, then transfer it to an evaporating dish, place it in a 45 °C water bath, repeatedly add anhydrous ethanol to remove trifluoroacetic acid and then dry. Add 0.5 mL each of 0.5 M 1-phenyl-3-methyl-5-pyrazolone (PMP) methanol solution and 0.3 M NaOH solution to the hydrolyzed monosaccharide sample. After complete dissolution, take 0.1 mL and react in a 70 °C water bath for 30 min for derivatization. After derivatization is completed, add 0.05 mL of 0.3 M hydrochloric acid, mix well, and then extract 3 times with chloroform to remove PMP. Transfer it through a 0.22 μm microporous filter membrane to a liquid phase bottle for detection.

[0092] Preparation of standard: Weigh 2 mg of the monosaccharide standard and directly perform derivatization. The treatment method is the same as that of the polysaccharide sample.

[0093] HPLC conditions: Use a Shimadzu HPLC system (LC-16 pump, SPD-M20A detector). The chromatographic column is Hypersil ODS2 C18 (4.6 mm × 250 mm, 5 μm). The mobile phase is 82% PBS (0.1 M, pH 7.0) and 18% acetonitrile (v / v). The flow rate is 1.0 mL·min -1 , the injection volume is 20 μL, and the detection wavelength is 245 nm.

[0094] Analyze the small molecular weight Fritillaria thunbergii polysaccharide obtained in Example 1 by gel permeation chromatography, and obtain its molecular weight distribution diagram as Figure 4 shown below.

[0095] The results show that the small molecular weight Fritillaria thunbergii polysaccharide in Example 1 contains galactose, arabinose, galacturonic acid, glucose and rhamnose, with a molecular weight > 50 kDa; the molar ratio of galactose, arabinose, galacturonic acid, and glucose is 31:19:12:35, and also contains a small amount of rhamnose (< 3%), mannose (< 0.5%) and glucuronic acid (< 0.5%). The small molecular weight Fritillaria thunbergii polysaccharide contains structural domains such as amyloid glucan, arabinogalactan, and polyrhamnogalacturonan. The spatial structure presents a combination of powdery and spherical structures. The molecular weight of the Fritillaria thunbergii polysaccharide nano-selenium complex > 50 kDa, and the monosaccharide composition is basically the same as that of the Fritillaria thunbergii polysaccharide.

[0096] Example 3

[0097] In vitro experiment to evaluate the antibacterial effect of the small molecule fritillaria thunbergii polysaccharide and its nano selenium complex obtained in Example 1. Experimental method: Take 30 g of agar, add 1.0 L of distilled water, dissolve it, place it in an autoclave for sterilization for 30 min, then make the sterilized culture medium into a plate. After solidification, drop 0.1 mL of the bacterial suspension with a concentration of 10 11 ~10 12 cfu / L, and spread it evenly. Prepare a series of concentration polysaccharide solutions of the small molecule fritillaria thunbergii polysaccharide and its nano selenium complex. Take 0.1 mL and add it into the Oxford cup, cover the petri dish, and culture it in an incubator at 37 °C for 24 h, and measure the diameter of the antibacterial circle formed in each Oxford cup.

[0098] Table 1 Inhibitory effects of different concentrations of small molecule fritillaria thunbergii polysaccharide

[0099]

[0100] Table 2 Inhibitory effects of different concentrations of fritillaria thunbergii polysaccharide nano selenium complex

[0101]

[0102] In this experiment, the outer diameter of the Oxford cup is 7.8 mm. Therefore, when the concentration of fritillaria thunbergii polysaccharide is lower than 1 mg / mL, and when the concentration of fritillaria thunbergii polysaccharide nano selenium complex is lower than 0.5 mg / mL, there is no inhibitory effect on the two tested strains. When the concentration of fritillaria thunbergii polysaccharide ≥ 2.5 mg / mL and the concentration of nano selenium complex ≥ 1 mg / mL, good antibacterial effects are shown on the two tested strains.

[0103] The above results show that the small molecular weight fritillaria thunbergii polysaccharide and its nano selenium complex can effectively inhibit bacterial proliferation, and the antibacterial effect of the fritillaria thunbergii polysaccharide nano selenium complex is better than that of the small molecular weight fritillaria thunbergii polysaccharide.

[0104] Example 4

[0105] Anti-inflammatory effect of fritillaria thunbergii polysaccharide and its nano-selenium complex, experimental method: Establishment of a rat model of pulmonary inflammation: On the 1st and 15th days, rats were anesthetized by intraperitoneal injection of 0.3% sodium pentobarbital solution, fixed in the supine position on the operating table, the skin in the middle of the neck was incised under sterile conditions, and after bluntly separating each layer of tissue, the trachea was exposed. The needle of a 1 mL syringe was inserted into the space between the tracheal cartilage rings of the rats, and 200 μL of 1 mg / mL LPS was slowly injected into the trachea. The normal control group was injected with an equal volume of normal saline under the same conditions, and the incision was sutured and disinfected. After injection, the rats were held upright and slowly rotated left and right for 3 min to evenly distribute the drug solution in the lungs. After waking up, they were routinely fed for 3 d, and then atomized and inhaled papain at 5 g / L, 2 mL for every 5 rats, 10 min each time, repeated 5 times every other day, and the model was established for 28 d. Fifty-six rats were randomly divided into 7 groups (8 rats in each group), namely the normal control group, the model group, the low-dose group of small-molecule fritillaria thunbergii polysaccharide (50 mg / kg), the medium-dose group (100 mg / kg), and the high-dose group (150 mg / kg). After the model was established, each group was given intragastric administration once a day for 20 consecutive days. The normal control group and the model group were given intragastric administration of an equal volume of normal saline. After the experiment, after anesthesia by intraperitoneal injection of 0.3% sodium pentobarbital solution, the chest cavity was opened along the trachea to separate the lung tissue, the left bronchus was ligated, 3 mL of normal saline was injected into the right lung through the bronchus, the lung was gently kneaded by hand for 30 s, the lavage fluid was aspirated, and the process was repeated 2 times. The lavage fluid was centrifuged at 1500 r / min for 15 min at 4 °C, and the supernatant was collected to detect the level of inflammatory factors, and the results were as Figure 5 shown. As Figure 5 can be seen, the levels of inflammatory factors IL-1β, IL-6, IL-17, and IFN in the model group all increased, and low, medium, and high concentrations of small-molecule fritillaria thunbergii polysaccharide could significantly reduce the levels of inflammatory factors.

[0106] The above results indicate that small-molecule fritillaria thunbergii polysaccharide can effectively regulate the levels of inflammatory factors in rats and has a certain anti-inflammatory effect.

[0107] Example 5

[0108] In a sterile environment, 28 g of sodium chloride, 110 g of citric acid, 110 g of sodium citrate, and 1.0 g of benzalkonium chloride were stirred and dissolved with 10 times the amount of pure water respectively. 50 g of the small-molecule fritillaria thunbergii polysaccharide obtained in Example 1 was stirred and dissolved with 1000 mL of pure water. After combining the above solutions, they were diluted to 10 L with pure water, filtered through a 0.5 μm filter membrane, and canned to obtain a small-molecule fritillaria thunbergii polysaccharide nasal spray.

[0109] Example 6

[0110] The formula of the body lotion is as follows: Oil phase: 5 parts of stearic acid, 2 parts of monoglyceryl stearate, 2 parts of isopropyl palmitate, 3 parts of petrolatum, 7 parts of white mineral oil, 1 part of cetyl alcohol, 0.3 part of essence, 0.1 part of methylparaben, 0.1 part of propylparaben; Aqueous phase: 5 parts of glycerol, 0.2 part of allantoin, 0.3 part of parachlorometaxylenol, 0.6 part of triethanolamine, 4 parts of fritillaria thunbergii polysaccharide nano-selenium complex obtained in Example 1, 69.4 parts of water.

[0111] Heat and stir to dissolve the oil phase (stearic acid, monoglyceryl stearate, isopropyl palmitate, petrolatum, white mineral oil, cetyl alcohol, methylparaben, propylparaben). Dissolve the fritillaria thunbergii polysaccharide nano-selenium complex in water, then add glycerol and triethanolamine, and heat and stir to dissolve. Then, slowly pour the aqueous phase into the oil phase under vigorous stirring. After treatment with a homogenizer, start heating and stirring. When cooled to 50 °C, add allantoin, parachlorometaxylenol and essence, and continue stirring and cooling to 30 °C for discharging. Let it stand for 2 - 3 days without change. After passing the inspection, it can be packaged.

[0112] Example 7

[0113] Take 1.2 kg of wolfberries and 0.8 kg of lilies, add 18 liters of water and bring to a boil, keep it simmering for 2 hours over a low heat, filter to obtain the filtrate, concentrate it to 10 liters, add 350 g of the low-molecular-weight fritillaria thunbergii polysaccharide obtained in Example 1, 850 g of honey, stir evenly, cool naturally, and let it stand for 3 hours to complete the precipitation. Use pressure filtration to obtain the filtrate, add 500 g of white sugar, 50 g of citric acid, and 3 g of salt, mix well, filter under pressure, and then filter through two-stage microporous membranes with pore sizes of 5 μm and 0.5 μm. The filtrate is pasteurized (85 °C, 50 min), and then it can be canned to obtain the low-molecular-weight fritillaria thunbergii polysaccharide oral liquid.

[0114] Example 8

[0115] Take 500 parts of water, 12 parts of sodium dodecyl sulfate, 10 parts of sodium sulfite, 14 parts of sodium chloride, 52 parts of AES, 5 parts of glycerol, 25 parts of coconut oil fatty acid diethanolamide, 30 parts of the fritillaria thunbergii polysaccharide nano-selenium complex obtained in Example 1. Place the water in a beaker, put it in a water bath at 45 °C, keep it at a constant temperature, add sodium dodecyl sulfate, sodium sulfite, and sodium chloride, stir to dissolve, then add AES, continue stirring to dissolve, add glycerol and coconut oil fatty acid diethanolamide. After the liquid becomes viscous, take out the beaker, cool it to room temperature, stir evenly, and divide it into portions to obtain the fritillaria thunbergii polysaccharide nano-selenium complex hand sanitizer.

[0116] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A small-molecular-weight fritillary bulb polysaccharide, characterized in that, It includes galactose, arabinose, galacturonic acid, glucose and rhamnose; the molecular weight of the small-molecular-weight Fritillaria thunbergii polysaccharide is 50 - 100 kDa; The preparation method of the small-molecular-weight Fritillaria thunbergii polysaccharide includes the following steps: S1) Mix Fritillaria thunbergii, complex enzyme and water, then heat for extraction and filter to obtain a filtrate; the complex enzyme includes papain, pectinase and amylase; S2) Concentrate the filtrate and centrifuge to obtain a supernatant; S3) Adjust the pH value of the supernatant to weakly acidic for acid hydrolysis reaction, and then adjust the pH value to weakly alkaline for alkaline hydrolysis reaction to obtain a hydrolysis solution; S4) Filter and dialyze the hydrolysis solution to obtain a dialysate; S5) Concentrate the dialysate and add ethanol for precipitation to obtain the small-molecular-weight Fritillaria thunbergii polysaccharide; In step S3), the pH value of the acid hydrolysis reaction is 5 - 6; the temperature of the acid hydrolysis reaction is 2°C - 6°C; the time of the acid hydrolysis reaction is 8 - 15 h; The pH value of the alkaline hydrolysis reaction is 7.8 - 8.5; the temperature of the alkaline hydrolysis reaction is 2°C - 6°C; the time of the alkaline hydrolysis reaction is 8 - 15 h; In step S3), use 0.01 - 0.5 mol / L trifluoroacetic acid aqueous solution to adjust the pH value of the supernatant to weakly acidic; Use 0.1 - 0.3 mol / L sodium bicarbonate aqueous solution to adjust the pH value to weakly alkaline.

2. The small-molecular-weight fritillaria thunbergii polysaccharide according to claim 1, wherein The molar ratio of galactose, arabinose, galacturonic acid and glucose is (28 - 34):(16 - 22):(10 - 14):(32 - 38).

3. A preparation method of the small molecular weight fritillaria thunbergii polysaccharide according to claim 1, characterized in that, It includes the following steps: S1) Mix Fritillaria thunbergii, complex enzyme and water, then heat for extraction and filter to obtain a filtrate; the complex enzyme includes papain, pectinase and amylase; S2) Concentrate the filtrate and centrifuge to obtain a supernatant; S3) Adjust the pH value of the supernatant to weakly acidic for acid hydrolysis reaction, and then adjust the pH value to weakly alkaline for alkaline hydrolysis reaction to obtain a hydrolysis solution; S4) Filter and dialyze the hydrolysis solution to obtain a dialysate; S5) Concentrate the dialysate and add ethanol for precipitation to obtain the small-molecular-weight Fritillaria thunbergii polysaccharide; In step S3), the pH value of the acid hydrolysis reaction is 5 - 6; the temperature of the acid hydrolysis reaction is 2°C - 6°C; the time of the acid hydrolysis reaction is 8 - 15 h; The pH value of the alkaline hydrolysis reaction is 7.8 - 8.5; the temperature of the alkaline hydrolysis reaction is 2°C - 6°C; the time of the alkaline hydrolysis reaction is 8 - 15 h; In step S3), use 0.01 - 0.5 mol / L trifluoroacetic acid aqueous solution to adjust the pH value of the supernatant to weakly acidic; Use 0.1 - 0.3 mol / L sodium bicarbonate aqueous solution to adjust the pH value to weakly alkaline.

4. The preparation method according to claim 3, characterized in that The mass ratio of papain, pectinase and amylase is 1:(2 - 4):(1 - 3); the mass ratio of Fritillaria thunbergii to the complex enzyme is (100 - 300):

1.

5. The preparation method according to claim 3, wherein In step S1), the temperature of the heat extraction is 50°C - 70°C; In step S2), concentrate to 1 / 20 - 1 / 5 of the volume of the filtrate; In step S4), the molecular weight cut-off of the membrane used for filtration and dialysis is 1200 - 1800 Da; In step S5), it is concentrated to 1 / 4 to 1 / 2 of the volume of the dialysis solution; the amount of ethanol added is such that the ethanol concentration in the system is 75% to 85%.

6. A fritillaria ussuriensis maxim polysaccharide nano-selenium complex, characterized in that, It is obtained by selenization of the small molecular weight fritillary bulb polysaccharide described in claim 1 or 2 or the small molecular weight fritillary bulb polysaccharide prepared by the preparation method described in any one of claims 3 to 5.

7. A preparation method of fritillaria ussuriensis polysaccharide nano-selenium complex, characterized in that, It includes the following steps: A1) Mix the small molecular weight fritillary bulb polysaccharide described in claim 1 or 2 or the small molecular weight fritillary bulb polysaccharide prepared by the preparation method described in any one of claims 3 to 5, sodium selenite and a reducing agent, and heat for a reduction reaction to obtain a nano selenium sol; A2) Remove the reducing agent from the nano selenium sol by high-speed centrifugation or dialysis to obtain a fritillary bulb polysaccharide nano selenium complex.

8. The preparation method according to claim 7, wherein The molar ratio of the sodium selenite to the reducing agent is 1:(3 to 25); the concentration of the small molecular weight fritillary bulb polysaccharide in the nano selenium sol is 300 to 1200 mg / L.

9. The preparation method according to claim 7, characterized in that, The temperature of the reduction reaction is 40°C to 85°C; the time of the reduction reaction is 0.2 to 5 h; The rotation speed of the high-speed centrifugation separation is 7500 to 12000 r / min; the time of the high-speed centrifugation is 10 to 35 min; The molecular weight cut-off of the dialysis membrane used for dialysis is 3 to 10000 kDa; the time of dialysis is 6 to 48 h.

10. The application of the small molecular weight fritillary bulb polysaccharide and / or the fritillary bulb polysaccharide nano selenium complex in the preparation of antibacterial and anti-inflammatory drugs and / or antibacterial and anti-inflammatory daily chemical products; The small molecular weight fritillary bulb polysaccharide is the small molecular weight fritillary bulb polysaccharide described in claim 1 or 2 or the small molecular weight fritillary bulb polysaccharide prepared by the preparation method described in any one of claims 3 to 5; The fritillary bulb polysaccharide nano selenium complex is the fritillary bulb polysaccharide nano selenium complex described in claim 6 or the fritillary bulb polysaccharide nano selenium complex prepared by the preparation method described in any one of claims 7 to 9.

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