A polygonum multiflorum starch-stilbene derivative complex, a preparation method and application thereof

Abstract

The present application provides a polygonum multiflorum starch-stilbene derivative complex, a preparation method and application thereof, and belongs to the technical field of starch complexes. The polygonum multiflorum starch is compounded with the stilbene derivative (or the polygonum multiflorum starch, polysaccharide and stilbene derivative are compounded), which is beneficial to improve the stability of the stilbene derivative. Specifically, after the polygonum multiflorum starch and the stilbene derivative form the complex, the amylose in the starch and the stilbene derivative are tightly wrapped, so that the hydrolysis of the digestive enzyme is slowed down, and the digestibility of the polygonum multiflorum starch is affected. The starch or starch + polysaccharide wrapped stilbene derivative can enhance the bioavailability of the stilbene derivative in the body. The polygonum multiflorum starch-stilbene derivative complex has a stable structure, the compounding degree is more than 80%, the release rate of the stilbene derivative in the simulated gastrointestinal fluid is effectively slowed down, and the biological activity of the stilbene derivative is better maintained.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of starch complexes, in particular to a polygonum multiflorum thunb starch-stilbene derivative complex and a preparation method and application thereof. BACKGROUND

[0002] Stilbene derivatives are a class of compounds with stilbene as the basic structure (such as stilbene glucoside, resveratrol, and polyphenols, etc.), which are widely present in plants and have various biological activities. In recent years, stilbene derivatives have attracted widespread attention in the fields of medicine, food, and cosmetics. Stilbene derivatives have been proven to have multiple pharmacological effects such as lowering blood lipids, anti-atherosclerosis, and preventing osteoporosis. Although stilbene derivatives have good biological activity, they have poor stability under conditions such as light, high temperature, alkaline medium, and iron ions due to their strong oxidizing property. Therefore, it is necessary to improve the stability of stilbene derivatives to maintain their biological activity and enhance their bioavailability. SUMMARY

[0003] Therefore, the present application aims to provide a polygonum multiflorum thunb starch-stilbene derivative complex and a preparation method and application thereof. The polygonum multiflorum thunb starch-stilbene derivative complex provided by the present application can improve the stability and bioavailability of stilbene derivatives and has good blood tonifying effect.

[0004] To achieve the above-mentioned application purposes, the present application provides the following technical solutions:

[0005] The present application provides a preparation method of a polygonum multiflorum thunb starch-stilbene derivative complex, comprising the following steps:

[0006] (1) providing polygonum multiflorum thunb starch;

[0007] (2) mixing the polygonum multiflorum thunb starch with stilbene derivatives and water to obtain a mixed emulsion; the stilbene derivatives include one or more of stilbene glucoside, resveratrol, and polyphenols;

[0008] (3) performing steam heating and drying on the mixed emulsion to obtain a polygonum multiflorum thunb starch-stilbene derivative complex.

[0009] Preferably, the step (2) is:

[0010] mixing the polygonum multiflorum thunb starch with stilbene derivatives, polysaccharides, and water to obtain a mixed emulsion.

[0011] Preferably, the mass of the stilbene glucoside is 1-10% of the mass of the polygonum multiflorum thunb starch.

[0012] Preferably, the mass of the stilbene glucoside is 1-10% of the mass of the polygonatum sibiricum red. The mass of the polysaccharide is 10-50% of the mass of the polygonatum sibiricum red.

[0013] Preferably, the preparation method of the polygonatum sibiricum red comprises the following steps:

[0014] The polygonatum sibiricum red powder is mixed with inorganic alkali liquor to perform a precipitation reaction, and the obtained precipitate is washed with water and centrifuged, and the white layer is the polygonatum sibiricum red.

[0015] The mass concentration of the inorganic alkali liquor is 0.02-0.2%.

[0016] Preferably, the polysaccharide comprises polygonatum sibiricum polysaccharide.

[0017] The polygonatum sibiricum polysaccharide is prepared by water extraction and alcohol precipitation.

[0018] Preferably, the temperature of the steam heating is 100-200 DEG C, and the time is 8-24 h.

[0019] Preferably, after the step (3), the obtained polygonatum sibiricum red-stilbene derivative complex is further subjected to post-treatment, and the post-treatment comprises:

[0020] The polygonatum sibiricum red-stilbene derivative complex is mixed with an alcohol solvent to perform centrifugation, and the lower layer precipitate after centrifugation is dried to obtain the purified polygonatum sibiricum red-stilbene derivative.

[0021] The polygonatum sibiricum red-stilbene derivative complex prepared by the preparation method is provided.

[0022] The polygonatum sibiricum red-stilbene derivative complex is provided for the preparation of blood tonifying drugs.

[0023] The polygonatum sibiricum red-stilbene derivative complex is provided for the preparation of blood tonifying drugs.

[0024] Polygonum multiflorum Thunb has the characteristic of "powdery enough", that is, the starch content is relatively high, but the research on the starch of Polygonum multiflorum Thunb is less. The starch of Polygonum multiflorum Thunb is compounded with stilbene derivatives (or the starch of Polygonum multiflorum Thunb, polysaccharide and stilbene derivatives are compounded), which is beneficial to improve the stability of stilbene derivatives. Specifically, after the starch of Polygonum multiflorum Thunb is compounded with stilbene derivatives, the amylose (or amylose + polysaccharide) in the starch is tightly wrapped with stilbene derivatives, so as to slow down the hydrolysis of digestive enzymes and reduce the digestibility of the starch of Polygonum multiflorum Thunb. The starch or starch + polysaccharide wrapping stilbene derivatives can enhance the bioavailability of stilbene derivatives in the body.

[0025] In the present application, from the molecular structure, stilbene derivatives have a destructive effect on the structure of the starch of Polygonum multiflorum Thunb. The main reasons for this phenomenon are two-fold. On the one hand, a large number of hydroxyl groups in stilbene derivatives can interact with water molecules in the starch of Polygonum multiflorum Thunb, retaining more water molecules; in the later drying process, more water molecules are evaporated, which is easy to form a more compact structure of the compound. On the other hand, stilbene derivatives can interact with starch chains through hydrogen bonds, hydrophobic and other non-covalent interactions, hinder the ordered interaction between starch chains, destroy the integrity of the surface of starch chains, make stilbene derivatives enter the interior of starch particles, and promote the starch to wrap stilbene derivatives, so as to increase the stability of stilbene derivatives and increase the bioavailability in the body. Therefore, the starch of Polygonum multiflorum Thunb-stilbene derivative compound obtained by the present application has a stable structure, the compounding degree is more than 80%, the release rate of stilbene derivatives in simulated gastrointestinal fluid is effectively slowed down, and the bioactivity of stilbene derivatives is better maintained.

[0026] The present application adopts the mode of steam heating to compound the starch of Polygonum multiflorum Thunb (or the starch of Polygonum multiflorum Thunb + polysaccharide) with stilbene derivatives, which can simulate the processing process of the medicinal material of Polygonum multiflorum Thunb itself and ensure the efficacy of the obtained compound. Meanwhile, steam heating also has the advantages of simplicity, high efficiency, green environmental protection and strong applicability.

[0027] In addition, the preparation method provided by the present application has the advantages of simple operation, short processing period, green environmental protection and strong applicability. The starch of Polygonum multiflorum Thunb-stilbene derivative compound obtained by the present application has good blood tonifying effect and can significantly improve the blood cell level and organ index, so it has high market promotion value.

[0028] Further, the preparation raw materials used in the present application, the starch of Polygonum multiflorum Thunb, stilbene glycoside and Polygonum multiflorum Thunb polysaccharide, are all derived from Polygonum multiflorum Thunb, which is beneficial to realize the resource utilization of Polygonum multiflorum Thunb. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1SEM 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;

[0030] Figure 2 XRD curves of the 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;

[0031] Figure 3 FTIR spectra of the 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;

[0032] Figure 4 Thermogravimetric analysis curves of the 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;

[0033] Figure 5 Results of the simulated gastrointestinal fluid digestion experiment 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;

[0034] Figure 6 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;

[0035] Figure 7 Blood supplementing 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. DETAILED DESCRIPTION

[0036] The present application provides a preparation method of a Polygonum multiflorum starch-stilbene derivative complex, comprising the following steps:

[0037] (1) providing Polygonum multiflorum starch;

[0038] (2) mixing the Polygonum multiflorum starch with a stilbene derivative and water to obtain a mixed emulsion; the stilbene derivative comprises one or more of stilbene glucoside, resveratrol and polydatin;

[0039] (3) steam heating and drying the mixed emulsion to obtain a Polygonum multiflorum starch-stilbene derivative complex.

[0040] In the present application, the preparation method of the Polygonum multiflorum starch preferably comprises the following steps:

[0041] The polygonum multiflorum powder is mixed with inorganic alkali liquor to carry out precipitation reaction, and the obtained precipitate is washed with water and centrifuged, and the white layer obtained is the polygonum multiflorum starch.

[0042] In the present application, the polygonum multiflorum powder is preferably from the rhizome part of polygonum multiflorum. In the present application, the particle size of the polygonum multiflorum powder is preferably 200 mesh.

[0043] In the present application, the inorganic alkali liquor is preferably one or more of sodium hydroxide solution, potassium hydroxide solution, sodium carbonate solution and sodium bicarbonate solution, and more preferably sodium hydroxide solution. In the present application, the mass concentration of the inorganic alkali liquor is preferably 0.02-2%, and more preferably 0.1-1%; the mass of the polygonum multiflorum powder to the volume of the inorganic alkali liquor is preferably 1:4. In the present application, the mixing method is preferably to soak the polygonum multiflorum starch in the inorganic alkali liquor. In the present application, the precipitation reaction is preferably carried out at room temperature under static conditions, and the time of the precipitation reaction is preferably 8-12h, and more preferably 10h. In the present application, the inorganic alkali liquor can destroy the cell wall and cell membrane of plant cells, so that the starch in the cells is released. At the same time, the inorganic alkali in the inorganic alkali liquor can also make the starch granules swell, increase its solubility in water, and facilitate subsequent separation and purification.

[0044] In the present application, the water used for washing is preferably deionized water, and the number of washing is preferably 3 times. In the present application, the speed of centrifugation is preferably 3000-4000 rpm, and the time is preferably 10-15 min. After centrifugation, the supernatant and the upper non-white layer containing cell wall are removed, and the white layer obtained is the polygonum multiflorum starch layer.

[0045] After obtaining the white layer, the present application resuspends the obtained white layer in distilled water, and sequentially carries out centrifugation, washing, drying and sieving to obtain purified polygonum multiflorum starch.

[0046] In the present application, the speed of centrifugation is preferably 3000 rpm, and the number of centrifugation is preferably 3 times. In the present application, the reagent used for washing is preferably ethanol; the drying temperature is preferably 30-40℃, and the time is preferably overnight. In the present application, the sieving is preferably 200 mesh.

[0047] The invention mixes the polygonum multiflorum starch with a stilbene derivative, water to obtain a mixed emulsion. In the invention, the stilbene derivative preferably includes one or several of stilbene glucoside, resveratrol and polydatin, preferably stilbene glucoside. In the invention, the mass of the stilbene glucoside is preferably 1-10% of the mass of the polygonum multiflorum starch, more preferably 2-8%, further preferably 3-5%. As a specific embodiment of the invention, the mass ratio of the polygonum multiflorum starch to water is preferably 3g:10mL. The invention does not have special requirements for the mixing method, and the mixing method known to those skilled in the art can be used, such as stirring mixing.

[0048] Alternatively, the invention mixes the polygonum multiflorum starch with a stilbene derivative, polysaccharide and water to obtain a mixed emulsion. In the invention, the stilbene derivative preferably includes one or several of stilbene glucoside, resveratrol and polydatin, preferably stilbene glucoside. In the invention, the mass of the stilbene glucoside is preferably 1-10% of the mass of the polygonum multiflorum starch, more preferably 2-8%, 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%, which can be 10%, 20%, 30%, 40% or 50%.

[0049] In the invention, the polysaccharide is preferably polygonum multiflorum polysaccharide. In the 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:

[0050] Boil the polygonum multiflorum medicinal material with water, mix the obtained supernatant with ethanol to obtain an alcohol precipitation solution;

[0051] Place the alcohol precipitation solution, remove the supernatant, and evaporate the ethanol in the precipitate to obtain a residual precipitate;

[0052] Mix the residual precipitate with hot water and freeze-dry to obtain polygonum multiflorum polysaccharide.

[0053] In the invention, the polygonum multiflorum medicinal material is preferably the rhizome part of polygonum multiflorum. In the invention, the mass ratio of the polygonum multiflorum medicinal material to water is preferably 1:8. In the invention, the polygonum multiflorum medicinal material is boiled with water for 2 times, and the single boiling time is preferably 1-3h. In the invention, the volume ratio of the supernatant to ethanol is preferably 1:2-4, more preferably 1:3.

[0054] In the invention, the temperature of the standing is preferably 4℃, and the time is preferably 8-12h; the method of evaporating the ethanol in the precipitate is preferably room temperature evaporation.

[0055] In the present application, the temperature of the hot water is preferably 100℃. The present application does not have special requirements for the freeze-drying method, and the freeze-drying method well known to those skilled in the art can be used.

[0056] After obtaining the mixed emulsion, the present application performs steam heating and drying on the mixed emulsion to obtain the polygonum multiflorum starch-stilbene derivative complex. In the present application, the temperature of the steam heating is preferably 100-200℃, more preferably 120-180℃, and specifically can be 100℃, 120℃, 140℃, 160℃, 180℃ or 200℃, and the time is preferably 8-24h, more preferably 12-20h, and specifically can be 8h, 12h, 16h, 20h or 24h.

[0057] In the present application, the temperature of the drying is preferably 30-40℃, more preferably 35℃; the present application does not have special requirements for the time of the drying, and the solid is dried to a constant weight.

[0058] In the present application, after the drying, the present application also preferably includes post-treatment of the obtained polygonum multiflorum starch-stilbene derivative complex, and the post-treatment includes:

[0059] The polygonum multiflorum starch-stilbene derivative complex is mixed with an alcohol solvent, centrifuged, and the lower precipitate after centrifugation is dried to obtain the purified polygonum multiflorum starch-stilbene derivative.

[0060] In the present application, the alcohol solvent is preferably ethanol; the mass of the polygonum multiflorum starch-stilbene derivative complex to the volume of the ethanol is preferably 1:20. In the present application, the rate of the centrifugation is preferably 1000rpm, and the time is preferably 5 minutes. The present application can remove the excess stilbene derivative in the polygonum multiflorum starch-stilbene derivative complex by mixing with the alcohol solvent and centrifugation. In the present application, the drying is preferably room temperature drying.

[0061] The present application provides the polygonum multiflorum starch-stilbene derivative complex prepared by the above preparation method. In the present application, the polygonum multiflorum starch-stilbene derivative complex is a complex of polygonum multiflorum starch and stilbene derivative, or a complex of polygonum multiflorum starch, polysaccharide and stilbene derivative.

[0062] The present application provides the application of the above polygonum multiflorum starch-stilbene derivative complex in the preparation of blood tonifying drugs.

[0063] The polygonum multiflorum starch-stilbene derivative complex, the preparation method and the application thereof provided by the present application will be described in detail below in conjunction with examples, but they should not be understood as limiting the scope of protection of the present application.

[0064] Example 1

[0065] (1) Preparation of Polygoni Multiflori Radix starch, using the following steps:

[0066] Polygoni Multiflori Radix was crushed and passed through a 200-mesh sieve. 200 g of the powder was soaked in 1600 mL of a 0.02% NaOH solution overnight. After the starch was precipitated, the supernatant was removed, and the precipitate was washed in deionized water to obtain a slurry containing starch.

[0067] The slurry containing starch was centrifuged at 3000 rpm for 10 minutes in a wide-mouth cup to remove the supernatant and the upper non-white layer containing cell walls. The white layer (starch layer) was resuspended in distilled water and centrifuged three more times and washed with ethanol. The resulting starch sample was dried overnight at 30-40°C, collected, and passed through a 200-mesh sieve to obtain Polygoni Multiflori Radix starch, denoted as S.

[0068] (2) Preparation method of Polygoni Multiflori Radix starch-diphenystilbene glucoside complex, using the following steps:

[0069] To 3 g of Polygoni Multiflori Radix starch, 4 wt% of diphenystilbene glucoside (TSG) based on the dry weight of the starch was added. The mixed sample was then dispersed in 10 mL of distilled water to obtain a mixed emulsion.

[0070] The mixed emulsion was placed in high-temperature steam at 100°C for 8 h and dried at 30-40°C for 8 h, crushed, ground, and passed through a 65-mesh sieve to obtain a Polygoni Multiflori Radix starch-diphenystilbene glucoside complex, denoted as ST.

[0071] Example 2

[0072] (1) Preparation of Polygoni Multiflori Radix polysaccharide, using the following steps:

[0073] 100 g of Polygoni Multiflori Radix was boiled in 800 mL of water twice for 1 hour each time. The supernatant was added to 4 times the volume of anhydrous ethanol, and the mixture was placed in a 4°C refrigerator for 12 hours. The supernatant was discarded, and the precipitate was dried. The precipitate was dissolved in hot water at 80°C and freeze-dried to obtain Polygoni Multiflori Radix polysaccharide.

[0074] (2) Preparation method of Polygoni Multiflori Radix starch-diphenystilbene glucoside complex, using the following steps:

[0075] To 3 g of Polygoni Multiflori Radix starch obtained in Example 1, 50 wt% of Polygoni Multiflori Radix polysaccharide based on the dry weight of the starch and 4 wt% of TSG based on the dry weight of the starch were added. The mixed sample was then dispersed in 20 mL of distilled water to obtain a mixed emulsion.

[0076] The mixed emulsion was placed in high-temperature steam at 100°C for 8 h and dried at 30-40°C for 8 h, crushed, ground, and passed through a 65-mesh sieve to obtain a Polygoni Multiflori Radix starch-diphenystilbene glucoside-Polygoni Multiflori Radix polysaccharide complex, denoted as STP.

[0077] Performance testing

[0078] (1) The degree of complexation of the polygonum multiflorum starch-stilbene glucoside complex and the polygonum multiflorum starch-stilbene glucoside-polygonum multiflorum polysaccharide complex

[0079] The method for testing the degree of complexation is performed using the following steps:

[0080] The degree of complexation (CI) of the polygonum multiflorum starch-stilbene glucoside complex was determined using the heating-refluxing-extraction method with ethanol as the solvent. 0.2 g of the polygonum multiflorum starch-stilbene glucoside complex was accurately weighed and placed in a conical flask with a stopper. Then, 25 mL of ethanol was added and heated to reflux for 30 minutes. After cooling, the supernatant was taken, filtered, and transferred to a syringe bottle. Then, the sample was analyzed on an Agilent 1260 high-performance liquid chromatography system (Agilent, San Jose, CA, USA) equipped with a quaternary pump, an automatic sampler, a constant-temperature column chamber, and a diode array detector. The separation was performed using an Agilent SB C18 chromatographic column (4.6 mm x 250 mm, 5 μm). The mobile phase was water (75%) and acetonitrile (25%) at a flow rate of 1.0 mL / min. The detection wavelength was 320 nm, and the injection volume was 10 µL. Using TSG as the standard, the content of free TSG in the complex was calculated. The degree of complexation (CI) = 100 x (M0-M1) / M0; where M0 represents the content of TSG originally added, and M1 represents the content of TSG extracted by ethanol. Each sample was run three times, and the average result was reported.

[0081] The degree of complexation 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 is shown in Table 1.

[0082] Table 1. Results of the degree of complexation test

[0083]

[0084] As can be seen from Table 1, the complex degree of the polygonum multiflorum starch-stilbene glucoside complex obtained in Example 1 is 95.39%, indicating that the polygonum multiflorum starch-stilbene glucoside complex is well formed, and the crosslinking of stilbene glucoside and starch in the complex system is almost saturated. The complex degree of the polygonum multiflorum starch-stilbene glucoside-polygonum multiflorum polysaccharide complex obtained in Example 2 is 80.69%, indicating that the complex is well formed. However, the complex degree of the polygonum multiflorum starch-stilbene glucoside-polygonum multiflorum polysaccharide complex is slightly lower than that of the polygonum multiflorum starch-stilbene glucoside complex, which may be due to the more complex structure of the polysaccharide, the larger molecular weight and functional group size. These characteristics may interfere with the interaction between the polysaccharide and the starch and stilbene glucoside.

[0085] (2) SEM study of polygonum multiflorum starch-stilbene glucoside complex and polygonum multiflorum starch-stilbene glucoside-polygonum multiflorum polysaccharide complex

[0086] The morphology of the polygonum multiflorum starch-stilbene glucoside complex and the polygonum multiflorum starch-stilbene glucoside-polygonum multiflorum polysaccharide complex in Examples 1 and 2 was characterized by scanning electron microscopy, and the results are shown in Figure 1 Figure 1 In the figure, A is polygonum multiflorum starch, B is stilbene glucoside, C is polygonum multiflorum starch-stilbene glucoside complex, D is polygonum multiflorum starch-stilbene glucoside-polygonum multiflorum polysaccharide complex, and a, b, c, d are the corresponding magnified images of A, B, C, D.

[0087] It can be seen that the morphology of the polygonum multiflorum starch-stilbene glucoside-polygonum multiflorum polysaccharide complex is neither the same as the original smooth spherical particles of polygonum multiflorum starch (A, a in Figure 1 ), nor the rod-like morphology of stilbene glucoside (B, b in Figure 1 ​B, b) in FIG. 1A, but present a blocky structure with wrinkled surface. The morphology of the polygonum multiflorum starch-stilbene glycoside complex changed greatly, the starch granules and rod-like TSG disappeared, the starch swelled and gelatinized, forming a block with holes on the surface. This shows that during the heating process, the starch gelatinizes, wraps TSG, and forms a complex. The polygonum multiflorum starch-stilbene glycoside complex forms a lump, showing a reticular structure, indicating that the starch absorbs water at high temperature, expands in volume, and causes the original granular structure to be destroyed. The results show that TSG has a destructive effect on the structure of the original starch. There are two main reasons for this phenomenon. On the one hand, the large number of hydroxyl groups in TSG can interact with water molecules in the polygonum multiflorum starch-stilbene glycoside complex, retaining more water molecules. Therefore, during the later drying process, more water molecules evaporate, forming a more compact structure. On the other hand, TSG can interact with starch chains through hydrogen bonds, hydrophobic and other non-covalent interactions, hinder the ordered interaction between starch chains, destroy the integrity of the surface of the starch chain, and cause the appearance of some fragments and irregular cavities. Although the polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex in FIG. 1C, Figure 1 D, d) in FIG. 1A is similar to the morphology of the polygonum multiflorum starch-stilbene glycoside complex in FIG. 1B, Figure 1 C, c) in FIG. 1A, and upon magnification, the structure of the polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex is more compact.

[0088] (3) XRD study of the polygonum multiflorum starch-stilbene glycoside complex and the polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex

[0089] Figure 2 XRD curves of the 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. Figure 2 In FIG. 2, S represents polygonum multiflorum starch, ST represents the polygonum multiflorum starch-stilbene glycoside complex, STP represents the polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex, and T represents TSG.

[0090] The formation of the starch-lipid complexes was confirmed by X-ray diffraction. The typical B-type crystal structure was observed for the Rhei starch, with the main diffraction peaks at 15°, 17°, 22° and 24° (2q). Compared with the Rhei starch, the Rhei starch-stilbene glycoside complex had very strong diffraction peaks at 10° and 20° (2q), showing a typical V-type structure, which also confirmed the formation of the complex. For the same starch, the relative crystallinity increased with the increase of the TSG content, indicating that the Rhei starch and TSG formed more ordered and stable crystals through hydrogen bonding. Unlike the Rhei starch-stilbene glycoside complex, the Rhei starch-stilbene glycoside-Rhei polysaccharide complex was formed by the interaction of starch, polysaccharide and stilbene glycoside. In this complexing process, the starch not only formed a complex with stilbene glycoside, but also formed a separate complex with polysaccharide. This was mainly because the heating process destroyed the structure of the Rhei starch molecules and the hydrogen bonds within the molecular chain, promoting the interaction of the Rhei starch and TSG.

[0091] (4) FTIR study of the Rhei starch-stilbene glycoside complex and the Rhei starch-stilbene glycoside-Rhei polysaccharide complex

[0092] Figure 3 FTIR spectra of the Rhei starch, TSG, the Rhei starch-stilbene glycoside complex obtained in Example 1 and the Rhei starch-stilbene glycoside-Rhei polysaccharide complex obtained in Example 2. Compared with stilbene glycoside and Rhei starch, the -OH stretching vibration in the Rhei starch-stilbene glycoside complex and the Rhei starch-stilbene glycoside-Rhei polysaccharide complex was red-shifted, indicating the formation of hydrogen bonds. At 1600 cm -1 , a broad weak absorption peak was observed, which could be attributed to the stretching vibration of the trans double bond (CH=CH) in stilbene glycoside; while the broad weak absorption peak at 2930 cm -1 was related to the asymmetric stretching vibration of the C-H group in the starch backbone. With the formation of the Rhei starch-stilbene glycoside complex and the Rhei starch-stilbene glycoside-Rhei polysaccharide complex, these two absorption signals were red-shifted to 1630 cm -1 and 2935 cm -1 , respectively, indicating the presence of hydrophobic interactions between stilbene glycoside and starch. Therefore, the FT-IR data showed that hydrogen bonding and hydrophobic interactions played a key role in promoting the formation of the Rhei starch-stilbene glycoside complex and the Rhei starch-stilbene glycoside-Rhei polysaccharide complex.

[0093] (5) TGA study of the Rhei starch-stilbene glycoside complex and the Rhei starch-stilbene glycoside-Rhei polysaccharide complex

[0094] Figure 4 Why the heat analysis curve 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, Figure 4 In the figure, A is the TGA curve and B is the DTG curve. The TGA curve shows that the first stage of weight loss peaks appears between 30℃ and 200℃, which is mainly due to the evaporation of free water and crystal water in the starch and the complex. The second stage of weight loss peaks appears between 200℃ and 360℃, which is mainly due to the degradation process of the starch and the guest molecules. The peak value on the DTG curve represents the maximum weight loss rate, and the peak temperature reflects the thermal stability of the sample. As can be seen from the DTG curve, the decomposition temperature of stilbene glycoside is 304.5℃, the thermal decomposition temperature of polygonum multiflorum starch is 318.3℃, and the decomposition temperature of the polygonum multiflorum starch-stilbene glycoside complex and the polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex is slightly higher, which is 321.8℃ and 324.9℃ respectively, which indicates that the thermal stability of the polygonum multiflorum starch-stilbene glycoside complex has been improved. This result is consistent with the subsequent stability experiment results, indicating that the stability of stilbene glycoside has been significantly improved, which is of great significance. The third stage of mass loss from 360℃ to 700℃ is due to the thermal degradation of carbon residue.

[0095] (6) Simulated gastrointestinal fluid digestion experiment of polygonum multiflorum starch-stilbene glycoside complex and polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex

[0096] The present application studies the release behavior of ST and STP in simulated gastric fluid (SGF, ingredients are 2.0 g NaCl, 10.0 g pepsin and 7.0 mL concentrated HCl, pH=1.2) and simulated intestinal fluid (SIF, ingredients are 10.0 g trypsin, 5.0 g pig bile salt, 6.0 g KH2PO4 and 0.2 M NaOH, pH=6.8). T (stilbene glycoside), ST and STP are respectively put into SGF and SIF, and incubated for 10 h under magnetic stirring. Then aliquots are taken at different time intervals. The release amount of T in the supernatant is determined by high performance liquid chromatography, and the release rate is calculated according to formula (1). All experiments are repeated 3 times.

[0097] Release rate (%) = W1 / W0×100% Formula (1);

[0098] In formula (1), W1 and W0 are respectively the release amount of T in the supernatant and the total amount of T in the ST and STP complex.

[0099] The simulated gastrointestinal fluid digestion experiment results 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 followsFigure 5 The results are shown in Figure 6. Figure 5 In the figure, A is the release behavior in simulated gastric fluid, and B is the release behavior in simulated intestinal fluid. As can be seen, after 8 hours of digestion in simulated intestinal fluid, only about 50% of free tetrahydroxystilbene glucoside remains, while about 60% of tetrahydroxystilbene glucoside in the polygonum multiflorum starch-tetrahydroxystilbene glucoside complex remains, and about 70% of tetrahydroxystilbene glucoside in the polygonum multiflorum starch-tetrahydroxystilbene glucoside-polygonum multiflorum polysaccharide complex remains. In simulated gastric fluid, after 8 hours of dissolution, the remaining amount of free tetrahydroxystilbene glucoside is 80%, while about 90% of tetrahydroxystilbene glucoside in the polygonum multiflorum starch-tetrahydroxystilbene glucoside complex remains, and about 95% of tetrahydroxystilbene glucoside in the polygonum multiflorum starch-tetrahydroxystilbene glucoside-polygonum multiflorum polysaccharide complex remains. These results show that the degradation rate of tetrahydroxystilbene glucoside in simulated intestinal fluid is significantly higher than that in simulated gastric fluid. Previous studies have also reached similar conclusions: tetrahydroxystilbene glucoside is relatively stable in acidic environments, but extremely unstable in alkaline environments. Therefore, the polygonum multiflorum starch-tetrahydroxystilbene glucoside-polygonum multiflorum polysaccharide complex has stronger stability than the polygonum multiflorum starch-tetrahydroxystilbene glucoside complex, which indicates that tetrahydroxystilbene glucoside is better encapsulated in the complex. This result shows that the polygonum multiflorum starch-tetrahydroxystilbene glucoside-polygonum multiflorum polysaccharide complex is relatively stable in gastrointestinal fluid, can preferentially release in the stomach environment, thereby effectively prolonging the residence time of tetrahydroxystilbene glucoside in the gastrointestinal fluid and promoting its absorption in the stomach.

[0100] (7) In vitro digestion experiment of polygonum multiflorum starch-tetrahydroxystilbene glucoside complex

[0101] The contents of fast digestible starch, slow digestible starch and resistant starch in the polygonum multiflorum starch and the polygonum multiflorum starch-tetrahydroxystilbene glucoside complex obtained in Example 1 were tested as follows:

[0102] First, 0.2 g (wet basis) of the test substance was suspended in 2 mL of distilled water and soaked in a boiling water bath for 20 minutes to ensure that the starch was completely gelatinized before enzymatic hydrolysis. The glucose content in the hydrolysate was determined at a specific time point using a GOPOD kit.

[0103] The contents of fast digestible starch (RDS), slow digestible starch (SDS) and resistant starch (RS) were calculated as shown in equations (2) to (4):

[0104] RDS (%) = (G 20 –G0)×0.9×100 equation (2);

[0105] SDS (%) = (G 120 –G 20 )×0.9×100 equation (3);

[0106] RS (%) = 100%–RDS (%)–SDS (%) equation (4);

[0107] wherein G0, G 20 , G 120 are the free glucose content, the glucose content released after 20 min, the glucose content released after 120 min, respectively.

[0108] The results obtained are shown in Table 2.

[0109] Table 2 Content of rapidly digestible starch, slowly digestible starch and resistant starch in polygonum multiflorum starch and polygonum multiflorum starch-stilbene glycoside complex

[0110]

[0111] As shown in Table 2, the content of rapidly digestible starch, slowly digestible starch and resistant starch in polygonum multiflorum starch was 38.29%, 16.04% and 45.66%, respectively. The digestibility of polygonum multiflorum starch-stilbene glycoside complex was affected by the content of stilbene glycoside, which showed that the content of rapidly digestible starch was significantly reduced, the content of slowly digestible starch increased by 17.05%, and the content of resistant starch increased by 10.65%. This significantly increased the resistance of starch to digestion. The content of slowly digestible starch in polygonum multiflorum starch-stilbene glycoside complex was 33.09%. The resistance to enzymes was attributed to the helical structure of polygonum multiflorum starch-stilbene glycoside complex, which prevented the dispersion of amylose and the combination of amylose and enzymes. The greater the swelling power, the higher the digestibility. Polygonum multiflorum starch-stilbene glycoside complex formed a highly stable microcrystalline structure, reducing the contact between starch molecules and water molecules, thereby inhibiting the hydration and swelling of amylopectin molecules. This would further hinder the entry of digestive enzymes into the starch molecules, thereby weakening the enzymatic action and greatly reducing the digestibility of starch.

[0112] (8) Pharmacokinetic study of polygonum multiflorum starch-stilbene glycoside complex and polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex

[0113] The pharmacokinetic study of polygonum multiflorum starch-stilbene glycoside complex and polygonum multiflorum starch-stilbene glycoside-polygonum multiflorum polysaccharide complex was carried out as follows:

[0114] Twelve adult male SD rats were randomly selected and divided into three groups, four rats in each group, namely T group, ST group and STP group. Before the experiment, the animals were kept in cages and not allowed to eat, but could drink water freely for 12 hours. Each 200 g rat was given 75 mg T, ST (containing 75 mg T) or STP (containing 75 mg T). Subsequently, 300 μL of orbital venous plexus blood was taken at 5, 15, 30 minutes and 1, 2, 4, 8, 12, 24 hours after administration, and placed in an edta-coated tube. The plasma was separated by centrifugation at 3500 rpm for 10 min, and was prepared for HPLC-QQQ-MS analysis.

[0115] The blood concentration-time curves of the F. ulmicaulis starch-stilbene glycoside complex of Example 1 and the F. ulmicaulis starch-stilbene glycoside-F. ulmicaulis polysaccharide complex of Example 2 in vivo are shown in FIG. 2, and the pharmacokinetic parameters are shown in Table 3. The pharmacokinetic parameters include the time of maximum concentration (T Figure 6 max ), the maximum concentration (C max ), the area under the concentration-time curve (AUC), and the mean residence time (MRT).

[0116] Table 3. Pharmacokinetic parameters of stilbene glycoside, F. ulmicaulis starch-stilbene glycoside complex, and F. ulmicaulis starch-stilbene glycoside-F. ulmicaulis polysaccharide complex (n = 6)

[0117]

[0118] The results show that, after oral administration of the F. ulmicaulis starch-stilbene glycoside complex, the plasma concentration reached a peak of 181.438 ± 7.109 ng / mL at 1 hour; and after oral administration of the F. ulmicaulis starch-stilbene glycoside-F. ulmicaulis polysaccharide complex, the plasma concentration reached a peak of 356.384 ± 105.674 ng / mL at 4 hours. The prolongation of the residence time of stilbene glycoside can be attributed to the sustained-release properties of F. ulmicaulis starch and F. ulmicaulis polysaccharide. Compared with administration of stilbene glycoside alone, the C max of the F. ulmicaulis starch-stilbene glycoside complex and the F. ulmicaulis starch-stilbene glycoside-F. ulmicaulis polysaccharide complex were 1.15 and 1.96 times higher, respectively, and the AUC （0-t） were 1.04 and 4.46 times higher, respectively. In summary, the pharmacokinetic study results show that F. ulmicaulis starch and F. ulmicaulis polysaccharide can prolong the pharmacological effect of stilbene glycoside and significantly improve its bioavailability in vivo. These findings show that the formation of the complex not only significantly enhances the bioavailability of stilbene glycoside, but also shows that the stability of the F. ulmicaulis starch-stilbene glycoside-F. ulmicaulis polysaccharide complex is superior to that of the F. ulmicaulis starch-stilbene glycoside complex, which is consistent with the results of the in vitro simulated gastrointestinal fluid experiment and has important application significance.

[0119] (9) Blood supplementing effect of the F. ulmicaulis starch-stilbene glycoside complex and the F. ulmicaulis starch-stilbene glycoside-F. ulmicaulis polysaccharide complex

[0120] The blood supplementing effect of the F. ulmicaulis starch-stilbene glycoside complex and the F. ulmicaulis starch-stilbene glycoside-F. ulmicaulis polysaccharide complex was tested as follows:

[0121] ​Thirty male Balb / c mice, 6 weeks old and weighing 20±2 g, were housed at a temperature controlled at 24±2℃ and a relative humidity maintained at 55%±10%. Animal experiments were conducted in accordance with relevant regulations for the care and use of laboratory animals. After one week of acclimatization, the 50 mice were randomly divided into five groups: control group, model group, T group, ST group, and STP group. The control and model groups received an equal volume of physiological saline via intragastric gavage. The medication was administered once every 24 hours for 9 consecutive days. On days 2 and 5, the model group, T group (0.1 g / kg), ST group (2.6 g / kg), and STP group (3.85 g / kg) were subcutaneously injected with acetylphenylhydrazine (20 mg / kg and 40 mg / kg), respectively. Starting from day 5, cyclophosphamide (40 mg / kg) was injected intraperitoneally one hour before medication, once daily for 4 consecutive days. The control group received an equal volume of physiological saline at the same time.

[0122] 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 As shown. Figure 7 In the table, A represents the white blood cell index, B represents the red blood cell index, C represents the hemoglobin index, D represents the thymus index, and E represents the spleen index.

[0123] Figure 7 Figures A through C show that, compared to the control group, all the above parameters in the model group were significantly lower (p<0.01), indicating successful model establishment. After treatment with stilbene glycoside, Polygonum multiflorum starch-stilbene glycoside complex, and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex, the levels of white blood cells, red blood cells, and hemoglobin concentration in each treatment group were significantly higher than those in the model group, with the complexes showing better efficacy. Organ coefficients for each group are shown below. 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, the organ indices of stilbene glycoside, Polygonum multiflorum starch-stilbene glycoside complex, and Polygonum multiflorum starch-stilbene glycoside-Polygonum multiflorum polysaccharide complex were all increased, with the organ indices of the complex group being significantly increased (p<0.05). This indicates that the Polygonum multiflorum starch-stilbene glycoside complex has a good blood-tonifying effect.

[0124] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered 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, Includes the following steps: (1) Provide Polygonum multiflorum starch; (2) The Polygonum multiflorum starch is mixed with stilbene derivative and water to obtain a mixed emulsion; the stilbene derivative includes stilbene glycoside; (3) The mixed emulsion was steam-heated and dried to obtain a Polygonum multiflorum starch-stilbene derivative complex; The mass of the stilbene glycoside is 1-10% of the mass of Polygonum multiflorum starch. The steam heating temperature is 100~200℃, and the time is 8~24h.

2. A method for preparing a Polygonum multiflorum starch-stilbene derivative complex, characterized in that, Includes the following steps: (1) Provide Polygonum multiflorum starch; (2) The Polygonum multiflorum starch is mixed with stilbene derivative, polysaccharide and water to obtain a mixed emulsion; the stilbene derivative includes stilbene glycoside; the polysaccharide includes Polygonum multiflorum polysaccharide; (3) The mixed emulsion was steam-heated and dried to obtain a Polygonum multiflorum starch-stilbene derivative complex; The mass of the stilbene glycoside is 1-10% of the mass of Polygonum multiflorum starch, and the mass of the polysaccharide is 10-50% of the mass of Polygonum multiflorum starch. The steam heating temperature is 100~200℃, and the time is 8~24h.

3. The preparation method according to claim 1 or 2, characterized in that, The method for preparing Polygonum multiflorum starch includes the following steps: Polygonum multiflorum powder was mixed with inorganic alkaline solution to carry out a precipitation reaction. The precipitate was washed with water and centrifuged. The white layer obtained was Polygonum multiflorum starch. The mass concentration of the inorganic alkaline solution is 0.02~0.2%.

4. The preparation method according to claim 2, characterized in that, The Polygonum multiflorum polysaccharide was prepared by water extraction and alcohol precipitation.

5. The preparation method according to claim 1 or 2, characterized in that, Following step (3), the process further includes post-treatment of the obtained Polygonum multiflorum starch-stilbene derivative complex, the post-treatment including: The Polygonum multiflorum starch-stilbene derivative complex was mixed with an alcohol solvent, centrifuged, and the lower precipitate after centrifugation was dried to obtain the purified Polygonum multiflorum starch-stilbene derivative.

6. The Polygonum multiflorum starch-stilbene derivative complex prepared by the preparation method according to any one of claims 1 to 5.

7. The use of the Polygonum multiflorum starch-stilbene derivative complex according to claim 6 in the preparation of blood-tonifying drugs.

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