Polygonum multiflorum starch-stilbene derivative compound as well as preparation method and application thereof

Abstract

The invention provides a polygonum multiflorum starch-stilbene derivative compound as well as a preparation method and application thereof, and belongs to the technical field of starch compounds. The fleece-flower root starch and the stilbene derivative are compounded (or fleece-flower root starch, polysaccharide and the stilbene derivative are compounded), the stability of the stilbene derivative is improved, specifically, after the fleece-flower root starch and the stilbene derivative form a compound, amylose in the starch and the stilbene derivative are tightly wrapped, and the stability of the stilbene derivative is improved. Therefore, the hydrolysis of digestive enzyme is retarded, and the digestion performance of the polygonum multiflorum starch is influenced; and the stilbene derivative is wrapped by the starch or the starch and the polysaccharide, so that the bioavailability of the stilbene derivative in vivo is enhanced. The polygonum multiflorum starch-stilbene derivative compound has a stable structure, the compounding degree reaches 80% or above, the release rate of the stilbene derivative in simulated gastrointestinal fluid is effectively slowed down, and the biological activity of the stilbene derivative is better maintained.

Description

Technical Field

[0001] The invention relates to the technical field of starch complexes, and in particular to a Polygonum multiflorum starch-stilbene derivative complex, a preparation method thereof, and an application thereof. Background Art

[0002] Stilbene derivatives are a class of compounds based on stilbene (e.g., stilbene glycosides, resveratrol, and polydatin). They are widely found in plants and possess a variety of biological activities. In recent years, they have attracted significant attention in the pharmaceutical, food, and cosmetic fields. Stilbene derivatives have been shown to exhibit diverse pharmacological effects, including hypolipidemic effects, anti-atherosclerotic effects, and osteoporosis prevention. Although stilbene derivatives exhibit promising biological activities, they are highly oxidizing and exhibit poor stability under conditions such as light, high temperature, alkaline media, and iron ions. Therefore, improving their stability is crucial to maintain their biological activity and enhance their bioavailability. Summary of the Invention

[0003] In view of this, the object of the present invention is to provide a Polygonum multiflorum starch-stilbene derivative complex and its preparation method and application. The Polygonum multiflorum starch-stilbene derivative complex provided by the present invention can improve the stability and bioavailability of stilbene derivatives and has good blood-tonifying effect.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The present invention provides a method for preparing a Polygonum multiflorum starch-stilbene derivative complex, comprising the following steps: (1) Providing Polygonum multiflorum starch; (2) mixing the Polygonum multiflorum starch with a stilbene derivative and water to obtain a mixed emulsion; the stilbene derivative includes one or more of stilbene glycoside, resveratrol and polydatin; (3) Steam heating and drying the mixed emulsion to obtain a Polygonum multiflorum starch-stilbene derivative complex.

[0005] Preferably, the step (2) is: The polygonum multiflorum starch is mixed with a stilbene derivative, a polysaccharide and water to obtain a mixed emulsion.

[0006] Preferably, the mass of the stilbene glycoside is 1-10% of the mass of Polygonum multiflorum starch.

[0007] Preferably, the mass of the stilbene glycoside is 1-10% of the mass of the Polygonum multiflorum starch, and the mass of the polysaccharide is 10-50% of the mass of the Polygonum multiflorum starch.

[0008] Preferably, the preparation method of the Polygonum multiflorum starch comprises the following steps: Mixing Polygonum multiflorum powder with an inorganic alkali solution to carry out a precipitation reaction, washing the obtained precipitate with water and centrifuging it to obtain a white layer of Polygonum multiflorum starch; The mass concentration of the inorganic alkali solution is 0.02-0.2%.

[0009] Preferably, the polysaccharide includes Polygonum multiflorum polysaccharide; The Polygonum multiflorum polysaccharide is prepared by water extraction and alcohol precipitation method.

[0010] Preferably, the steam heating temperature is 100-200° C. and the time is 8-24 hours.

[0011] Preferably, after step (3), the obtained Polygonum multiflorum starch-stilbene derivative complex is subjected to post-treatment, and the post-treatment comprises: The polygonum multiflorum starch-stilbene derivative complex is mixed with an alcohol solvent, and the mixture is centrifuged. The lower layer precipitate after the centrifugation is dried to obtain a purified polygonum multiflorum starch-stilbene derivative.

[0012] The present invention provides a Polygonum multiflorum starch-stilbene derivative complex prepared by the above preparation method.

[0013] The present invention provides application of the above-mentioned Polygonum multiflorum starch-stilbene derivative complex in preparing blood-tonifying medicines.

[0014] The present invention provides a method for preparing a Polygonum multiflorum starch-stilbene derivative complex, comprising the following steps: (1) providing Polygonum multiflorum starch; (2) mixing the Polygonum multiflorum starch with a stilbene derivative and water to obtain a mixed emulsion, wherein the stilbene derivative comprises one or more of stilbene glycoside, resveratrol, and polydatin; and (3) steam heating and drying the mixed emulsion to obtain the Polygonum multiflorum starch-stilbene derivative complex. Alternatively, step (2) may also be: mixing the Polygonum multiflorum starch with a stilbene derivative, a polysaccharide, and water to obtain a mixed emulsion.

[0015] Polygonum multiflorum is known for its high starch content, but research on this starch is limited. The present invention combines Polygonum multiflorum starch with a stilbene derivative (or combines Polygonum multiflorum starch, a polysaccharide, and a stilbene derivative), which helps improve the stability of the stilbene derivative. Specifically, after the Polygonum multiflorum starch and the stilbene derivative form a complex, the amylose (or amylose + polysaccharide) in the starch tightly encapsulates the stilbene derivative, slowing hydrolysis by digestive enzymes and reducing the digestibility of the Polygonum multiflorum starch. Furthermore, encapsulating the stilbene derivative with the starch or starch + polysaccharide complex enhances its bioavailability in the body.

[0016] In the present invention, from the perspective of molecular structure, the diphenylethylene derivatives have a destructive effect on the structure of Polygonum multiflorum starch. There are two main reasons for this phenomenon. On the one hand, the large number of hydroxyl groups present in the diphenylethylene derivatives can interact with the water molecules in the Polygonum multiflorum starch, retaining more water molecules; in the later drying process, more water molecules are evaporated, which makes it easy for the complex to form a denser structure. On the other hand, the diphenylethylene derivatives can interact with the starch chains through non-covalent interactions such as hydrogen bonds and hydrophobicity, hindering the orderly interaction between the starch chains, destroying the integrity of the starch chain surface, allowing the diphenylethylene derivatives to enter the interior of the starch granules, prompting the starch to wrap around the diphenylethylene derivatives, increasing the stability of the diphenylethylene derivatives, and increasing the bioavailability in the body. Therefore, the Polygonum multiflorum starch-diphenylethylene derivative complex obtained by the present invention has a stable structure, a composite degree of more than 80%, effectively slowing down the release rate of the diphenylethylene derivatives in simulated gastrointestinal fluid, and better maintaining the biological activity of the diphenylethylene derivatives.

[0017] The present invention uses steam heating to compound Polygonum multiflorum starch (or Polygonum multiflorum starch + polysaccharide) with a stilbene derivative, mimicking the processing process of Polygonum multiflorum itself and ensuring the efficacy of the resulting compound. Steam heating also offers the advantages of simplicity, efficiency, and environmental friendliness.

[0018] In addition, the preparation method provided by the present invention is simple to operate, has a short processing cycle, is green and environmentally friendly, and has strong applicability. The obtained Polygonum multiflorum starch-stilbene derivative complex has a good blood-tonifying effect, can significantly improve blood cell levels and organ indexes, and has high market promotion value.

[0019] Furthermore, the raw materials used in the present invention, such as Polygonum multiflorum starch, stilbene glycosides, and Polygonum multiflorum polysaccharide, are all derived from Polygonum multiflorum, which is conducive to the resource utilization of Polygonum multiflorum. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 These are SEM images of the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1 and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2; Figure 2 XRD curves of Polygonum multiflorum starch, TSG, the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1, and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2; Figure 3 FTIR spectra of Polygonum multiflorum starch, TSG, the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1, and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2; Figure 4Thermogravimetric analysis curves of Polygonum multiflorum starch, TSG, the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1, and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2; Figure 5 These are the results of simulated gastrointestinal fluid digestion experiments on the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1 and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2; Figure 6 The blood concentration-time curves of the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1 and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2 in vivo; Figure 7 The blood-tonifying effects of the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1 and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2 are shown. DETAILED DESCRIPTION

[0021] The present invention provides a method for preparing a Polygonum multiflorum starch-stilbene derivative complex, comprising the following steps: (1) Providing Polygonum multiflorum starch; (2) mixing the Polygonum multiflorum starch with a stilbene derivative and water to obtain a mixed emulsion; the stilbene derivative includes one or more of stilbene glycoside, resveratrol and polydatin; (3) Steam heating and drying the mixed emulsion to obtain a Polygonum multiflorum starch-stilbene derivative complex.

[0022] In the present invention, the preparation method of Polygonum multiflorum starch preferably comprises the following steps: Mixing Polygonum multiflorum powder with an inorganic alkali solution to carry out a precipitation reaction, washing the obtained precipitate with water and centrifuging it to obtain a white layer of Polygonum multiflorum starch; In the present invention, the Polygonum multiflorum powder is preferably derived from the rhizome of Polygonum multiflorum. In the present invention, the particle size of the Polygonum multiflorum powder is preferably under 200 sieves.

[0023] In the present invention, the inorganic alkali solution is preferably one or more of sodium hydroxide solution, potassium hydroxide solution, sodium carbonate solution and sodium bicarbonate solution, more preferably sodium hydroxide solution. In the present invention, the mass concentration of the inorganic alkali solution is preferably 0.02~2%, more preferably 0.1~1%; the volume ratio of the mass of the Polygonum multiflorum powder to the inorganic alkali solution is preferably 1:4. In the present invention, the mixing method is preferably to soak the Polygonum multiflorum starch in the inorganic alkali solution. In the present invention, the precipitation reaction is preferably carried out at room temperature and under static conditions, and the precipitation reaction time is preferably 8~12h, more preferably 10h. In the present invention, the inorganic alkali solution can destroy the cell walls and cell membranes of plant cells, thereby releasing the starch in the cells. At the same time, the inorganic alkali in the inorganic alkali solution can also swell the starch granules, increase its solubility in water, and facilitate subsequent separation and purification.

[0024] In the present invention, the water used for the water washing is preferably deionized water, and the number of water washings is preferably three. In the present invention, the centrifugation speed is preferably 3000-4000 rpm, and the time is preferably 10-15 minutes. After the centrifugation, the present invention removes the supernatant and the upper non-white layer containing the cell walls, and the resulting white layer is the Polygonum multiflorum starch layer.

[0025] After obtaining the white layer, the present invention resuspends the obtained white layer in distilled water, and sequentially performs centrifugation, washing, drying and sieving to obtain purified Polygonum multiflorum starch.

[0026] In the present invention, the centrifugation speed is preferably 3000 rpm, and the number of centrifugation cycles is preferably 3 times. In the present invention, the washing agent is preferably ethanol; the drying temperature is preferably 30-40°C, and the drying time is preferably overnight. In the present invention, the sieving is preferably through a 200-mesh sieve.

[0027] The present invention mixes the Polygonum multiflorum starch with a diphenylethylene derivative and water to obtain a mixed emulsion. In the present invention, the diphenylethylene derivative preferably includes one or more of diphenylethylene glycosides, resveratrol and polydatin, preferably diphenylethylene glycosides. In the present invention, the mass of the diphenylethylene glycosides is preferably 1 to 10% of the mass of the Polygonum multiflorum starch, more preferably 2 to 8%, and further preferably 3 to 5%. As a specific embodiment of the present invention, the mass ratio of the Polygonum multiflorum starch to water is preferably 3g:10mL. The present invention has no special requirements for the mixing method, and a mixing method familiar to those skilled in the art can be used, such as stirring and mixing.

[0028] Alternatively, the present invention mixes the Polygonum multiflorum starch with a stilbene derivative, a polysaccharide and water to obtain a mixed emulsion. In the present invention, the stilbene derivative preferably includes one or more of stilbene glycosides, resveratrol and polydatin, preferably stilbene glycosides. In the present invention, the mass of the stilbene glycosides is preferably 1-10% of the mass of the Polygonum multiflorum starch, more preferably 2-8%, and further preferably 3-5%; the mass of the polysaccharide is preferably 10-50% of the mass of the Polygonum multiflorum starch, more preferably 20-40%, and can be 10%, 20%, 30%, 40% or 50%.

[0029] In the present invention, the polysaccharide is preferably Polygonum multiflorum polysaccharide. In the present invention, the Polygonum multiflorum polysaccharide is preferably prepared by water extraction and alcohol precipitation. Specifically, the preparation method of the Polygonum multiflorum polysaccharide preferably includes the following steps: Boil the Radix Polygoni Multiflori with water, and mix the obtained supernatant with ethanol to obtain an alcohol precipitate; The alcohol precipitation solution is allowed to stand, the supernatant is removed, and the ethanol in the precipitate is volatilized to obtain a residual precipitate; The residual precipitate is mixed with hot water and freeze-dried to obtain Polygonum multiflorum polysaccharide.

[0030] In the present invention, the Radix Polygoni Multiflori medicinal material is preferably the rhizome of Radix Polygoni Multiflori. In the present invention, the mass ratio of the Radix Polygoni Multiflori medicinal material to water is preferably 1:8. In the present invention, the Radix Polygoni Multiflori medicinal material is preferably boiled twice with water, and the single boiling time is preferably 1 to 3 hours. In the present invention, the volume ratio of the supernatant to ethanol is preferably 1:2 to 4, more preferably 1:3.

[0031] In the present invention, the standing temperature is preferably 4° C., and the standing time is preferably 8 to 12 hours; and the method of volatilizing the ethanol in the precipitate is preferably room temperature volatilization.

[0032] In the present invention, the temperature of the hot water is preferably 100° C. The present invention has no special requirements for the freeze-drying method, and any freeze-drying method well known to those skilled in the art can be used.

[0033] After obtaining the mixed emulsion, the present invention steam-heats and dries the mixed emulsion to obtain a Polygonum multiflorum starch-stilbene derivative complex. In the present invention, the steam heating temperature is preferably 100-200°C, more preferably 120-180°C, specifically 100°C, 120°C, 140°C, 160°C, 180°C, or 200°C, and the steam heating time is preferably 8-24 hours, more preferably 12-20 hours, specifically 8 hours, 12 hours, 16 hours, 20 hours, or 24 hours.

[0034] In the present invention, the drying temperature is preferably 30-40° C., more preferably 35° C.; the present invention has no special requirements for the drying time, and the solid can be dried to a constant weight.

[0035] In the present invention, after the drying, the present invention further preferably includes post-processing the obtained Polygonum multiflorum starch-stilbene derivative complex, and the post-processing includes: The polygonum multiflorum starch-stilbene derivative complex is mixed with an alcohol solvent, and the mixture is centrifuged. The lower layer precipitate after the centrifugation is dried to obtain a purified polygonum multiflorum starch-stilbene derivative.

[0036] In the present invention, the alcohol solvent is preferably ethanol; the ratio of the mass of the Polygonum multiflorum starch-stilbene derivative complex to the volume of ethanol is preferably 1:20. In the present invention, the centrifugation rate is preferably 1000 rpm, and the time is preferably 5 minutes. The present invention can remove excess stilbene derivatives from the Polygonum multiflorum starch-stilbene derivative complex by mixing with the alcohol solvent and centrifuging. In the present invention, the drying is preferably performed at room temperature.

[0037] The present invention provides a Polygonum multiflorum starch-stilbene derivative complex prepared by the above preparation method. In the present invention, the Polygonum multiflorum starch-stilbene derivative complex is a complex of Polygonum multiflorum starch and a stilbene derivative, or a complex of Polygonum multiflorum starch, a polysaccharide and a stilbene derivative.

[0038] The present invention provides application of the above-mentioned Polygonum multiflorum starch-stilbene derivative complex in preparing blood-tonifying medicines.

[0039] The following examples will describe in detail the Polygonum multiflorum starch-stilbene derivative complex provided by the present invention, its preparation method and application, but they should not be construed as limiting the scope of protection of the present invention.

[0040] Example 1 (1) Preparation of Polygonum multiflorum starch, using the following steps: The Polygonum multiflorum was ground and passed through a 200-mesh sieve. 200 g of the powder was soaked in 1600 mL of a 0.02% NaOH solution overnight to precipitate starch. The supernatant was removed and the precipitate was washed in deionized water to obtain a starch-containing slurry.

[0041] Centrifuge the starch-containing slurry at 3000 rpm in a wide-mouthed cup for 10 minutes. Remove the supernatant and the upper non-white layer containing the cell walls. Resuspend the white layer (starch layer) in distilled water, centrifuge three times, and wash with ethanol. Dry the resulting starch sample overnight at 30-40°C, collect it, and pass it through a 200-mesh sieve to obtain Polygonum multiflorum starch, designated as S.

[0042] (2) A method for preparing a Polygonum multiflorum starch-stilbene glycoside complex comprises the following steps: To 3 g of Polygonum multiflorum starch was added 4 wt% of stilbene glucoside (TSG) based on the dry weight of the starch, and the mixed sample was dispersed in 10 mL of distilled water to obtain a mixed emulsion.

[0043] The mixed emulsion was placed in a high-temperature steam heating at 100°C for 8 hours, dried at 30-40°C for 8 hours, crushed, ground, and passed through a 65-mesh sieve to obtain a Polygonum multiflorum starch-stilbene glycoside complex, which was marked as ST.

[0044] Example 2 (1) Preparation of Polygonum multiflorum polysaccharide, using the following steps: 100 g of Polygonum multiflorum was added to 800 mL of water and boiled twice for 1 hour each time. Four times the volume of anhydrous ethanol was added to the supernatant and the mixture was allowed to stand in a refrigerator at 4°C for 12 hours. The supernatant was discarded and the precipitate was evaporated to dryness, dissolved in 80°C hot water and freeze-dried to obtain Polygonum multiflorum polysaccharide.

[0045] (2) A method for preparing a Polygonum multiflorum starch-stilbene glycoside complex comprises the following steps: To 3 g of the Polygonum multiflorum starch obtained in Example 1, 50 wt % of the Polygonum multiflorum starch dry weight of Polygonum multiflorum polysaccharide and 4 wt % of the starch dry weight of TSG were added. The mixed sample was then dispersed in 20 mL of distilled water to obtain a mixed emulsion.

[0046] The mixed emulsion was placed in a high-temperature steam heating at 100°C for 8 hours, dried at 30-40°C for 8 hours, crushed, ground, and passed through a 65-mesh sieve to obtain a Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex, which was recorded as STP.

[0047] Performance Testing (1) Degree of complexation of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex The composite degree test method adopts the following steps: The free TSG content and the degree of complexation (CI) of the Polygonum multiflorum starch-stilbene glucoside complex were determined using a heating-reflux-extraction method using ethanol as the solvent. 0.2 g of the Polygonum multiflorum starch-stilbene glucoside complex was accurately weighed and placed in a stoppered Erlenmeyer flask. 25 mL of ethanol was then added and the mixture was extracted under heating and reflux for 30 minutes. After cooling, the supernatant was filtered and transferred to an injection vial. The sample was then analyzed on an Agilent 1260 HPLC system (Agilent, San Jose, CA, USA) equipped with a quaternary pump, autosampler, thermostatted column compartment, and diode array detector. Separation was performed on an Agilent SB C18 column (4.6 mm × 250 mm, 5 μm). The mobile phase consisted of water (75%) and acetonitrile (25%) at a flow rate of 1.0 mL / min. Detection was performed at a wavelength of 320 nm, and the injection volume was 10 μL. The free TSG content in the complex was calculated using TSG as a standard. Composite index (CI) = 100 × (M0 - M1) / M0, where M0 represents the amount of TSG added initially and M1 represents the amount of TSG extracted with ethanol. Each sample was run three times, and the average result was reported.

[0048] The complexation degrees of the Polygonum multiflorum starch-stilbene glucoside complex (ST) obtained in Example 1 and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex (STP) obtained in Example 2 are shown in Table 1.

[0049] Table 1 Composite degree test results

[0050] As can be seen from Table 1, the degree of complexation of the Polygonum multiflorum starch-stilbene glycoside complex obtained in Example 1 is 95.39%, indicating that the Polygonum multiflorum starch-stilbene glycoside complex is well formed and the cross-linking of stilbene glycoside and starch in the composite system is almost saturated. The degree of complexation of the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex obtained in Example 2 is 80.69%, indicating that the complex is well formed. However, the degree of complexation of the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex is slightly lower than that of the Polygonum multiflorum starch-stilbene glycoside complex, which may be due to the fact that the polysaccharide has a more complex structure and its molecular weight and functional group size are larger. These characteristics may interfere with the interaction between the polysaccharide and starch and stilbene glycoside.

[0051] (2) SEM study of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex The morphology of the Polygonum multiflorum starch-stilbene glycoside complex and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex in Examples 1 and 2 was characterized by scanning electron microscopy. The results are shown in FIG. Figure 1 shown. Figure 1 In the figure, A is Polygonum multiflorum starch, B is diphenyl glycoside, C is Polygonum multiflorum starch-diphenyl glycoside complex, D is Polygonum multiflorum starch-diphenyl glycoside-Polygonum multiflorum polysaccharide complex, and a, b, c, and d are the enlarged pictures corresponding to A, B, C, and D.

[0052] It can be seen that the morphology of the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex is different from the original smooth spherical particles of Polygonum multiflorum starch ( Figure 1 A, a), and also with the rod-shaped form of stilbene glycosides ( Figure 1 Unlike (B and b) in Figure 2, the structure exhibits a blocky, wrinkled surface. The morphology of the Polygonum multiflorum starch-stilbene glycoside complex undergoes significant changes. The starch granules and rod-shaped TSG disappear, and the starch swells and gelatinizes, forming a blocky structure with pores on the surface. This indicates that during heating, the starch gelatinizes, encapsulating the TSG and forming a complex. The Polygonum multiflorum starch-stilbene glycoside complex forms clumps and a network-like structure, indicating that the starch absorbs water at high temperatures, causing volume expansion and disruption of the original granular structure. These results indicate that TSG disrupts the native starch structure. Two main reasons contribute to this phenomenon. First, the numerous hydroxyl groups in TSG interact with water molecules in the Polygonum multiflorum starch-stilbene glycoside complex, retaining more water molecules. Consequently, more water molecules evaporate during the subsequent drying process, resulting in a denser structure. Second, TSG can interact with starch chains through non-covalent interactions such as hydrogen bonding and hydrophobic interactions, hindering the orderly interactions between starch chains and disrupting the integrity of the starch chain surface, leading to the appearance of fragments and irregular cavities. Although the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex ( Figure 1 The morphology of D, d) is similar to that of the Polygonum multiflorum starch-stilbene glycoside complex ( Figure 1 C, c in the figure) Magnified observation shows that the structure of the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex is relatively dense.

[0053] (3) XRD study of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex Figure 2 XRD curves of Polygonum multiflorum starch, TSG, the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1, and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2. Figure 2 In the table, S represents Polygonum multiflorum starch, ST represents Polygonum multiflorum starch-stilbene glucoside complex, STP represents Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex, and T represents TSG.

[0054] X-ray diffraction confirmed the formation of starch-lipid complexes. Polygonum multiflorum starch exhibits a typical B-type crystal structure, with major diffraction peaks at 15°, 17°, 22°, and 24° (2θ). Compared to Polygonum multiflorum starch, the Polygonum multiflorum starch-stilbene glycoside complex exhibits strong diffraction peaks at 10° and 20° (2θ), demonstrating a typical V-shaped structure. The appearance of this V-shaped structure also confirms the formation of the complex. For the same starch, the relative crystallinity increases with increasing TSG content, indicating that Polygonum multiflorum starch and TSG form more ordered and stable crystals through hydrogen bonding. Unlike the Polygonum multiflorum starch-stilbene glycoside complex, the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex is formed by the interaction of starch, polysaccharide, and stilbene glycoside. During this complexation process, starch can form complexes not only with stilbene glycoside but also with the polysaccharide itself. This is primarily because the heating process disrupts the molecular structure and intramolecular hydrogen bonds of Polygonum multiflorum starch, promoting the interaction between Polygonum multiflorum starch and TSG.

[0055] (4) FTIR study of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex Figure 3 FTIR spectra of Polygonum multiflorum starch, TSG, the Polygonum multiflorum starch-stilbene glycoside complex obtained in Example 1, and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex obtained in Example 2. Compared with stilbene glycoside and Polygonum multiflorum starch, the -OH stretching vibration in the Polygonum multiflorum starch-stilbene glycoside complex and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex red-shifted, indicating the formation of hydrogen bonds. -1 At 2930 cm -1 The broad weak absorption peaks appearing at 1630 cm-1 are related to the asymmetric stretching vibration of the CH group in the starch backbone. With the formation of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex, the two absorption signals red-shifted to 1630 cm-1, respectively. -1 and 2935 cm -1 , indicating that there is a hydrophobic interaction between stilbene glycosides and starch. Therefore, FT-IR data show that hydrogen bonding and hydrophobic interactions play a key role in promoting the formation of Polygonum multiflorum starch-stilbene glycoside complexes and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complexes.

[0056] (5) TGA study of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex Figure 4Thermogravimetric analysis curves of Polygonum multiflorum starch, TSG, the Polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1, and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex obtained in Example 2, Figure 4 In Figure 1, A is the TGA curve, and B is the DTG curve. The TGA curve shows that the first stage of weight loss peaks occurs between 30°C and 200°C, primarily due to the evaporation of free and crystalline water in the starch and the complex. The second stage of weight loss peaks occurs between 200°C and 360°C, primarily due to the degradation of starch and the guest molecule. The peak on the DTG curve represents the maximum weight loss rate, while the peak temperature reflects the thermal stability of the sample. The DTG curves show that the decomposition temperature of stilbene glycoside is 304.5°C, while the thermal decomposition temperature of Polygonum multiflorum starch is 318.3°C. However, the decomposition temperatures of the Polygonum multiflorum starch-stilbene glycoside complex and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex are slightly higher, at 321.8°C and 324.9°C, respectively. This indicates that the thermal stability of the Polygonum multiflorum starch-stilbene glycoside complex has been improved. This result is consistent with the results of subsequent stability experiments, indicating that the stability of stilbene glycoside has been significantly improved, which is of great significance. The third stage of mass loss from 360 °C to 700 °C is attributed to the thermal degradation of carbonaceous residue.

[0057] (6) Simulated gastrointestinal digestion experiments of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex The release behavior of ST and STP in simulated gastric fluid (SGF, composed of 2.0 g NaCl, 10.0 g pepsin, and 7.0 mL concentrated HCl, pH = 1.2) and simulated intestinal fluid (SIF, composed of 10.0 g trypsin, 5.0 g porcine bile salts, 6.0 g KH2PO4, and 0.2 M NaOH, pH = 6.8) was investigated. T (stilbene glycoside), ST, and STP were placed in SGF and SIF, respectively, and incubated under magnetic stirring for 10 h. Aliquots were then collected at different time intervals. The amount of T released in the supernatant was determined by high-performance liquid chromatography, and the release rate was calculated according to formula (1). All experiments were repeated three times.

[0058] Release rate (%) = W1 / W0×100% Formula (1); In formula (1), W1 and W0 are the released amount of T in the supernatant and the total amount of T in the ST and STP complexes, respectively.

[0059] The results of the simulated gastrointestinal digestion experiment on the Polygonum multiflorum starch-stilbene glycoside complex obtained in Example 1 and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex obtained in Example 2 are as follows: Figure 5 shown. Figure 5In the figure, A represents the release behavior in simulated gastric fluid, and B represents the release behavior in simulated intestinal fluid. It can be seen that after 8 hours of digestion in simulated intestinal fluid, only approximately 50% of the free stilbene glycosides remained, while approximately 60% of the stilbene glycosides in the Polygonum multiflorum starch-stilbene glycoside complex remained, and approximately 70% of the stilbene glycosides in the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex remained. After 8 hours of dissolution in simulated gastric fluid, the remaining amount of free stilbene glycosides was 80%, while approximately 90% of the stilbene glycosides in the Polygonum multiflorum starch-stilbene glycoside complex remained, and approximately 95% of the stilbene glycosides in the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex remained. These results indicate that the degradation rate of stilbene glycosides in simulated intestinal fluid is significantly higher than that in simulated gastric fluid. Previous studies have reached similar conclusions: stilbene glycosides are relatively stable in acidic environments but extremely unstable in alkaline environments. This shows that the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex has stronger stability than the Polygonum multiflorum starch-stilbene glycoside complex, indicating that the stilbene glycoside is better encapsulated in the complex. This result shows that the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex is relatively stable in gastrointestinal fluid and can be preferentially released in the gastric environment, thereby effectively prolonging the residence time of stilbene glycoside in the gastrointestinal fluid and promoting its absorption in the stomach.

[0060] (7) In vitro digestion experiment of Polygonum multiflorum starch-stilbene glycoside complex The contents of rapidly digestible starch, slowly digestible starch and resistant starch in the Polygonum multiflorum starch and the Polygonum multiflorum starch-stilbene glycoside complex obtained in Example 1 were tested as follows: First, 0.2 g (wet basis) of the test substance was suspended in 2 mL of distilled water and immersed in a boiling water bath for 20 minutes to ensure complete gelatinization of the starch before enzymatic hydrolysis. Glucose content in the hydrolyzate was determined at specific time points using a GOPOD kit.

[0061] The contents of rapidly digestible starch (RDS), slowly digestible starch (SDS) and resistant starch (RS) were calculated as shown in formulas (2) to (4): RDS (%) = (G 20 –G0)×0.9×100 Formula (2); SDS (%) = (G 120 –G 20 )×0.9×100 Formula (3); RS (%) = 100%–RDS (%)–SDS (%) Equation (4); Among them, G0, G 20 , G 120 They are free glucose content, glucose content released after 20 minutes, and glucose content released after 120 minutes.

[0062] The results are shown in Table 2.

[0063] Table 2 Contents of rapidly digestible starch, slowly digestible starch and resistant starch in Polygonum multiflorum starch and Polygonum multiflorum starch-stilbene glycoside complex

[0064] As shown in Table 2, the rapidly digestible starch, slowly digestible starch, and resistant starch contents of Polygonum multiflorum starch are 38.29%, 16.04%, and 45.66%, respectively. The digestibility of the Polygonum multiflorum starch-stilbene glycoside complex is affected by the stilbene glycoside content, with a significant decrease in the rapidly digestible starch content, an increase of 17.05% in the slowly digestible starch content, and a 10.65% increase in the resistant starch content. This significantly increases the starch's digestibility. The slowly digestible starch content of the Polygonum multiflorum starch-stilbene glycoside complex is 33.09%. This enzyme resistance is attributed to the helical structure of the Polygonum multiflorum starch-stilbene glycoside complex, which prevents the dispersion of amylose and the binding of amylose to enzymes. Greater swelling power is associated with higher digestibility. The Polygonum multiflorum starch-stilbene glycoside complex forms a highly stable microcrystalline structure, reducing the contact between starch molecules and water molecules, thereby inhibiting the hydration and swelling of amylopectin molecules. This further hinders the entry of digestive enzymes into the starch molecules, weakening enzymatic degradation and significantly reducing starch digestibility.

[0065] (8) Pharmacokinetic study of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex The pharmacokinetic study of the Polygonum multiflorum starch-stilbene glycoside complex and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex was conducted as follows: Twelve adult male Sprague-Dawley rats were randomly selected and divided into three groups of four: T, ST, and STP. Before the experiment, the animals were caged and not allowed to eat, but had free access to water for 12 hours. Each 200 g rat was administered 75 mg of T, ST (containing 75 mg of T), or STP (containing 75 mg of T). Subsequently, 300 μL of blood was collected from the orbital venous plexus at 5, 15, and 30 minutes, and 1, 2, 4, 8, 12, and 24 hours after administration and placed in EDTA-coated tubes. Plasma was separated by centrifugation at 3500 rpm for 10 minutes and prepared for HPLC-QQQ-MS analysis.

[0066] ② The blood concentration-time curves of the Polygonum multiflorum starch-stilbene glycoside complex obtained in Example 1 and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex obtained in Example 2 are as follows: Figure 6 Its pharmacokinetic parameters are shown in Table 3. Among them, the pharmacokinetic parameters include the maximum concentration time (Tmax ), maximum concentration (C max ), area under the concentration-time curve (AUC), and mean residence time (MRT).

[0067] Table 3 Pharmacokinetic parameters of stilbene glucoside, Polygonum multiflorum starch-stilbene glucoside complex, and Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex (n=6)

[0068] The results showed that after oral administration of Polygonum multiflorum starch-stilbene glycoside complex, the plasma concentration reached a peak of 181.438±7.109 ng / mL at 1 hour; while after oral administration of Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex, the plasma concentration reached a peak of 356.384±105.674 ng / mL at 4 hours. The prolonged retention time of stilbene glycoside can be attributed to the sustained-release properties of Polygonum multiflorum starch and Polygonum multiflorum polysaccharide. Compared with the administration of stilbene glycoside alone, the C max 1.15-fold and 1.96-fold higher, respectively, and AUC （0-t） The results of the pharmacokinetic study showed that Polygonum multiflorum starch and Polygonum multiflorum polysaccharide can prolong the pharmacological effects of stilbene glycosides and significantly improve their bioavailability in vivo. These findings indicate that the formation of the complex can not only significantly enhance the bioavailability of stilbene glycosides, but also show that the stability of the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex is superior to that of the Polygonum multiflorum starch-stilbene glycoside complex. This result is consistent with the results of the in vitro gastrointestinal fluid simulation experiment and has important application significance.

[0069] (9) The blood-tonifying effect of Polygonum multiflorum starch-stilbene glycoside complex and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex The blood-tonifying effects of the Polygonum multiflorum starch-stilbene glycoside complex and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex were tested as follows: Thirty six-week-old male Balb / c mice, weighing 20 ± 2 g, were housed in an environment maintained at a temperature of 24 ± 2°C and a relative humidity of 55% ± 10%. Animal experiments were conducted in accordance with the relevant regulations for the care and use of laboratory animals. After one week of adaptive feeding, the 50 mice were randomly divided into five groups: control, model, T, ST, and STP. The control and model groups received an equal volume of normal saline intragastrically every 24 hours for nine days. On days 2 and 5, the model, T, ST, and STP groups received subcutaneous injections of acetylphenylhydrazine (20 mg / kg and 40 mg / kg), respectively. Starting on day 5, cyclophosphamide (40 mg / kg) was administered intraperitoneally one hour before drug administration, once daily for four days. The control group received an equal volume of normal saline at the same time.

[0070] The blood-tonifying effects of the Polygonum multiflorum starch-stilbene glycoside complex obtained in Example 1 and the Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex obtained in Example 2 are as follows: Figure 7 shown. Figure 7 In the chart, A is the white blood cell index, B is the red blood cell index, C is the hemoglobin index, D is the thymus index, and E is the spleen index.

[0071] Figure 7 Figures A to C show that the above parameters in the model group were significantly lower than those in the control group (p < 0.01), indicating that the model was successfully established. After treatment with stilbene glycoside, Polygonum multiflorum starch-stilbene glycoside complex, and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex, the white blood cell, red blood cell, and hemoglobin concentrations of mice in each treatment group were significantly higher than those in the model group, and the complex was more effective. Figure 7 As shown in Figures D-E. Compared with the control group, the thymus and spleen indices of rats in the model group were significantly decreased (p < 0.05). Compared with the model group, organ indices of stilbene glucoside, the Polygonum multiflorum starch-stilbene glucoside complex, and the Polygonum multiflorum starch-stilbene glucoside-Polygonum multiflorum polysaccharide complex all increased, with the complex group showing a significant increase in organ indices (p < 0.05). This suggests that the Polygonum multiflorum starch-stilbene glucoside complex has a potent blood-tonifying effect.

[0072] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A method for preparing a Polygonum multiflorum starch-stilbene derivative complex, characterized in that: The following steps are involved: (1) Providing Polygonum multiflorum starch; (2) mixing the Polygonum multiflorum starch with a stilbene derivative and water to obtain a mixed emulsion; the stilbene derivative includes one or more of stilbene glycoside, resveratrol and polydatin; (3) Steam heating and drying the mixed emulsion to obtain a Polygonum multiflorum starch-stilbene derivative complex.

2. The preparation method according to claim 1, characterized in that The step (2) is: The polygonum multiflorum starch is mixed with a stilbene derivative, a polysaccharide and water to obtain a mixed emulsion.

3. The preparation method according to claim 1, characterized in that The mass of the stilbene glycosides is 1-10% of the mass of Polygonum multiflorum starch.

4. The preparation method according to claim 2, characterized in that The mass of the stilbene glycoside is 1-10% of the mass of the Polygonum multiflorum starch, and the mass of the polysaccharide is 10-50% of the mass of the Polygonum multiflorum starch.

5. The preparation method according to claim 1 or 2, characterized in that The preparation method of the Polygonum multiflorum starch comprises the following steps: Mixing Polygonum multiflorum powder with an inorganic alkali solution to carry out a precipitation reaction, washing the obtained precipitate with water and centrifuging it to obtain a white layer of Polygonum multiflorum starch; The mass concentration of the inorganic alkali solution is 0.02-0.2%.

6. The preparation method according to claim 2, characterized in that The polysaccharide includes Polygonum multiflorum polysaccharide; The Polygonum multiflorum polysaccharide is prepared by water extraction and alcohol precipitation method.

7. The preparation method according to claim 1 or 2, characterized in that The steam heating temperature is 100-200° C. and the time is 8-24 hours.

8. The preparation method according to claim 1 or 2, characterized in that After step (3), the obtained Polygonum multiflorum starch-stilbene derivative complex is subjected to post-treatment, and the post-treatment comprises: The polygonum multiflorum starch-stilbene derivative complex is mixed with an alcohol solvent, and the mixture is centrifuged. The lower layer precipitate after the centrifugation is dried to obtain a purified polygonum multiflorum starch-stilbene derivative.

9. The Polygonum multiflorum starch-stilbene derivative complex prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the Polygonum multiflorum starch-stilbene derivative complex according to claim 9 in the preparation of blood-tonifying medicines.

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