Qinba selenium mushroom extract FA-2-b-beta and extraction method

Abstract

The invention relates to the technical field of fungus extracts, in particular to a Qinba selenium mushroom extract FA-2-b-beta and an extraction method. The method comprises the following steps: decocting a Qinba selenium mushroom raw material for multiple times to obtain a crude extract, and performing alcohol-water extraction on the crude extract for multiple times; wherein the temperature of each time of decoction is 80 to 120 DEG C, and the time of each time of decoction is 0.5 to 2 hours; the alcohol is an ethanol solution with multiple volume percentages. According to the method, the traditional decoction treatment and alcohol-water extraction methods are effectively combined, so that the Qinba selenium mushroom extract FA-2-b-beta can be effectively extracted. The extraction method does not need to use a complex organic solvent for extraction, and has the advantages of simple steps, mild conditions, easiness in operation and low cost. According to the extraction method, extraction of the FA-2-b-beta is realized for the first time, and a good method is provided for subsequent research and application of the FA-2-b-beta.

Description

Technical Field

[0001] The present invention relates to the technical field of fungus extracts, and in particular to a Qinba selenium mushroom extract FA-2-b-β and an extraction method thereof. Background Art

[0002] Qinba selenium mushroom, also known as Agaricus blazei and Agaricus chinensis, belongs to the Basidiomycetes class and is rich in nutritional and medicinal value. It is a rare fungus that can be used as both medicine and food. The polysaccharides in Qinba selenium mushroom have a wide range of application prospects. Studies have confirmed that the polysaccharides extracted from Qinba selenium mushroom have the effects of inhibiting tumor cell proliferation, regulating tumor cell growth and aging, and improving the body's immunity.

[0003] However, in the prior art, the research on Qinba Selenium Mushroom mainly focuses on the selenium content therein, while the research on the polysaccharides contained therein is still relatively small. At the same time, there is no specific extraction method for the polysaccharide extracts in Qinba Selenium Mushroom. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a Qinba selenium mushroom extract FA-2-b-β and an extraction method.

[0005] The technical solution of the present invention to solve the above technical problems is as follows:

[0006] The invention provides a method for extracting FA-2-b-β of Qinba selenium mushroom extract. The method comprises the following steps: firstly subjecting Qinba selenium mushroom raw materials to multiple decoction treatments to obtain a crude extract, and then subjecting the crude extract to multiple alcohol-water extractions; wherein the decoction temperature each time is 80-120°C, and the decoction time each time is 0.5-2h; and the alcohol is an ethanol solution with various volume percentages.

[0007] Based on the above technical solution, the present invention can also be improved as follows.

[0008] Further, the following steps are included:

[0009] S1, obtaining the Qinba selenium mushroom raw material, adding water and decocting for 2 to 5 times, filtering and combining the decoctions obtained after each decoction, and then concentrating to obtain the crude extract;

[0010] S2, adding a first ethanol solution to the crude extract, mixing and filtering to obtain a first precipitate;

[0011] S3, mixing the first precipitate with water and centrifuging for multiple times, and mixing the supernatants obtained from each centrifugation to obtain a first mixed solution;

[0012] S4, dialyzing the first mixed solution to obtain a dialysate;

[0013] S5, adding a second ethanol solution to the dialysate, mixing, standing and filtering to obtain a second precipitate;

[0014] S6. Wash the second precipitate with a third ethanol solution and then dry it to obtain the Qinba selenium mushroom extract acidic RNA protein complex FA-2-b-β.

[0015] Further, the volume percentage of ethanol in the first ethanol solution is less than the volume percentage of ethanol in the second ethanol solution, and the volume percentage of ethanol in the second ethanol solution is less than the volume percentage of ethanol in the third ethanol solution.

[0016] Further, in step S2, the volume percentage of ethanol in the first ethanol solution is 75%.

[0017] Furthermore, in step S2, the volume ratio of the crude extract to the first ethanol solution is 1:7-9.

[0018] Furthermore, in step S5, the volume percentage of the second ethanol solution is 90-95%.

[0019] Further, in step S5, the volume ratio of the dialysate to the second ethanol solution is 1:8-9.

[0020] Furthermore, in step S6, the third ethanol solution is anhydrous ethanol.

[0021] Furthermore, the Qinba selenium mushroom raw material is Qinba selenium mushroom powder, and the mesh number of the Qinba selenium mushroom powder is 80-120 meshes.

[0022] The present invention also provides a Qinba selenium mushroom extract FA-2-b-β, which is extracted by the above method. The structural formula of the Qinba selenium mushroom extract FA-2-b-β is shown in formula (1):

[0023]

[0024] The beneficial effects of the present invention are:

[0025] The extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention effectively combines the traditional decoction treatment and the alcohol-water extraction method, so that the Qinba selenium mushroom extract FA-2-b-β can be effectively extracted. The extraction method does not need to use a complex organic solvent for extraction, has simple steps, mild conditions, and has the advantages of easy operation and low cost. Since FA-2-b-β has multiple functions such as anti-tumor, immune regulation, and health care, the extraction method realizes the extraction of the effective ingredient of FA-2-b-β for the first time and determines its chemical formula, which provides a good method and experimental basis for the subsequent research and application of FA-2-b-β. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The total ion current diagram of the D-generation sample obtained in Example 3 is the extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention;

[0027] Figure 2 The total ion current diagram of the H-generation sample obtained in Example 3 is the extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention;

[0028] Figure 3 The extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention, the obtained 1 H-NMR spectrum;

[0029] Figure 4 The extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention, the obtained 13 C-NMR spectrum;

[0030] Figure 5 The COSY spectrum obtained in Example 4 is the extraction method of FA-2-b-β of the Qinba selenium mushroom extract of the present invention;

[0031] Figure 6 The NOESY spectrum obtained in Example 4 is the extraction method of FA-2-b-β of the Qinba selenium mushroom extract of the present invention;

[0032] Figure 7 The HSQC spectrum obtained in Example 4 is the extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention;

[0033] Figure 8 This is the HMBC spectrum obtained in Example 4 of the extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention. DETAILED DESCRIPTION

[0034] The principles and features of the present invention are described below in conjunction with the accompanying drawings. The examples given are only used to explain the present invention and are not used to limit the scope of the present invention.

[0035] The extraction method of the Qinba selenium mushroom extract FA-2-b-β of the present invention comprises the following steps: firstly subjecting the Qinba selenium mushroom raw material to multiple decoction treatments to obtain a crude extract, and then subjecting the crude extract to multiple alcohol-water extractions; wherein the temperature of each decoction is 80-120°C, and the time of each decoction is 0.5-2h; and the alcohol is an ethanol solution with various volume percentages.

[0036] The extraction method of the FA-2-b-β of the Qinba selenium mushroom extract of the present invention effectively combines the traditional decoction treatment and the alcohol-water extraction method, so that the FA-2-b-β of the Qinba selenium mushroom extract can be effectively extracted. Among them, the decoction temperature and decoction time are specifically for the Qinba selenium mushroom, which can effectively extract the FA-2-b-β therein, and multiple extractions are combined to achieve a good extraction effect.

[0037] The method of the present invention comprises the following steps:

[0038] S1. Obtain Qinba selenium mushroom raw material, add water and boil for 2 to 5 times, filter the decoctions obtained after each boiling respectively, combine them, and then concentrate them to obtain a crude extract.

[0039] Preferably, the decoction temperature is 100° C. each time, and the decoction time is 1 hour each time.

[0040] S2. Add the first ethanol solution to the crude extract, mix and filter to obtain a first precipitate.

[0041] Preferably, in the first ethanol solution, the volume percentage of ethanol is 75%.

[0042] Preferably, the volume ratio of the crude extract to the first ethanol solution is 1:7-9; further preferably, in the mixture of the crude extract and the first ethanol solution, the volume percentage of ethanol is 60%-65%.

[0043] S3. Mix the first precipitate with water and centrifuge for multiple times, and mix the supernatants obtained from each centrifugation to obtain a first mixed solution.

[0044] S4. Dialyze the first mixed solution to obtain a dialysate.

[0045] S5. Add the second ethanol solution to the dialysate, mix, let stand and filter to obtain a second precipitate.

[0046] Preferably, the volume percentage of the second ethanol solution is 90-95%.

[0047] Preferably, the volume ratio of the dialysate to the second ethanol solution is 1:8-9; further preferably, in the mixture of the dialysate and the second ethanol solution, the volume percentage of ethanol is 80%-85%.

[0048] S6. Wash the second precipitate with a third ethanol solution and then dry it to obtain the Qinba selenium mushroom extract acidic RNA protein complex FA-2-b-β.

[0049] Preferably, the third ethanol solution is anhydrous ethanol.

[0050] Preferably, the Qinba selenium mushroom raw material used in the present invention is Qinba selenium mushroom powder, and the mesh number of the Qinba selenium mushroom powder is 80-120 meshes.

[0051] The Qinba selenium mushroom extract FA-2-b-β of the present invention has been experimentally verified to have a structural formula as shown in formula (1):

[0052]

[0053] The present invention is described in detail below through specific examples.

[0054] Example 1

[0055] This example uses the method of the present invention to extract FA-2-b-β, and the specific steps are:

[0056] S1. Obtain Qinba selenium mushroom powder with a mesh size of 100 mesh, add water and boil for 3 times, filter the decoctions obtained after each boiling respectively, combine them, and then concentrate them to obtain a crude extract. The temperature of each boiling is 100° C., and the time of each boiling is 1 hour.

[0057] S2, adding the first ethanol solution to the crude extract, mixing and filtering to obtain a first precipitate. The volume percentage of ethanol in the first ethanol solution is 75%, and the volume percentage of ethanol in the mixed solution of the crude extract and the first ethanol solution is 65%.

[0058] S3. Mix the first precipitate with water and centrifuge for multiple times, and mix the supernatants obtained from each centrifugation to obtain a first mixed solution.

[0059] S4. Dialyze the first mixed solution to obtain a dialysate.

[0060] S5, adding a second ethanol solution to the dialysate, mixing, standing and filtering to obtain a second precipitate. The volume percentage of the second ethanol solution is 95%, and the volume percentage of ethanol in the mixed solution of the dialysate and the second ethanol solution is 85%.

[0061] S6. Wash the second precipitate with anhydrous ethanol and then dry it to obtain an extract containing FA-2-b-β.

[0062] Example 2 Preliminary verification of the extract

[0063] (1) The extract obtained in Example 1 was dissolved in water, a sample to be tested was prepared, and its optical rotation was measured. After extraction and measurement by the method of the present invention, the optical rotation of the extract obtained in Example 1 was [a]D-23 (c1.00, 0.3M NaOH), containing (1-6)-β-Dglucopyranan. After the sample to be tested was purified by affinity chromatography, ultraviolet spectrum analysis was performed, and the test results showed that: characteristic absorption peaks were located at 228.5nm and 255.8nm, indicating that it contained protein and nucleic acid substances, wherein the molecular weight of the nucleic acid was 1000, and the base components were A, Ap, Cp, Up and other modified bases.

[0064] Among them, the affinity chromatography method is UPLC-MS / MS analysis, and the specific process is as follows:

[0065] a. Loading: Load the purified and concentrated extract onto the affinity chromatography column.

[0066] b. Elution: The eluent is used to elute the chromatographic column to achieve separation based on the difference in affinity between the target component and the ligand.

[0067] c. Collect the eluate: Collect the eluate containing the target component during the elution process.

[0068] d. Preliminary analysis: Preliminary analysis of the components in the eluate is performed by UV spectrophotometry, mass spectrometry, etc.

[0069] Chromatographic conditions: Water Acquity UPLCHSS T3 Co l umn (2.1mm×100mm, 1.8μm). Mobile phase with injection temperature of 25℃, acetonitrile (A) and 0.1% formic acid aqueous solution (B). Elution gradient: 0%-95% A, flow rate: 0.3mL / min, injection volume: 5μL.

[0070] Mass spectrometry conditions: ES I ion source; m / z scanning range: 100-1000; capillary voltage: 4000 V, drying gas flow rate: 10 L / min, drying gas temperature: 350 °C, nebulization pressure: 30 psi, decomposition voltage: 80 V, collision energy: 90 V, and data were collected using Xcalibur 4.0 software.

[0071] The specific process of UV spectrum analysis is:

[0072] a. Place the sample solution in a cuvette, place it in the instrument light path, and start the scanning program to obtain the spectrum.

[0073] b. Observe spectral characteristics: observe the shape, peak position, peak shape and other characteristics of the spectrum to determine the functional groups or structural information that may exist in the sample.

[0074] c. Calculate absorbance: Measure the absorbance value at the characteristic wavelength for subsequent qualitative identification and quantitative determination.

[0075] d. Identify the ingredients in traditional Chinese medicine samples based on ultraviolet spectral characteristics.

[0076] d-1. Compare with standard spectrum: Compare the UV spectrum of the sample with the standard spectrum of the known component to determine whether the sample contains the component.

[0077] d-2. Characteristic peak identification: Based on the position, shape and other information of the characteristic peak, further confirm the type of components in the sample.

[0078] d-3. Establish a standard curve: Use a series of standard solutions of known concentrations to perform spectral scanning and draw a standard curve.

[0079] d-4. Sample determination: Determine the absorbance value of the sample solution under the same conditions and calculate the content of the target component based on the standard curve.

[0080] Example 3 Polysaccharide Methylation Analysis

[0081] Methylation analysis is one of the most powerful means of structural analysis of polysaccharides and oligosaccharides. It can obtain partially methylated sugar alcohol acetyl derivatives, and use gas chromatography-mass spectrometry (GC-MS) for qualitative and quantitative analysis to determine the connection mode of each monosaccharide residue. The experimental methods for derivatization of different types of polysaccharides are different, in order to more accurately analyze the type of sugar residues in the sample. The polysaccharide samples are divided into two categories: neutral sugars and acidic sugars, so as to ensure the accuracy of the polysaccharide bonding structure results. This project mainly uses GC-MS to analyze the polysaccharide bonding structure.

[0082] The methylation analysis of this embodiment is as follows:

[0083] (1) Sample pretreatment

[0084] Weigh about 5 mg of the extract obtained in Example 1, add 1 ml of pure water to dissolve, add 1 ml of 100 mg / ml 1-cyclohexyl-2-morpholineethylcarbodiimide methyl p-toluenesulfonate, and react for 2 hours. Add 1 ml of 2M imidazole, divide the sample into two parts, add 1 ml of 30 mg / ml NaBH4 and 1 ml of 30 mg / ml NaBD4 respectively, and react for 3 hours.

[0085] Add 100μ glacial acetic acid to terminate the reaction. After dialysis for 48h, freeze-dry the sample. Add 500μ DMSO to the sample to dissolve, add 1mg NaOH, incubate for 30min, and add 50μ iodomethane solution to react for 1h.

[0086] Add 1ml water and 2ml dichloromethane, vortex to mix, centrifuge, and discard the water phase. Repeat the water wash 3 times. Aspirate the lower dichloromethane phase and blow dry with nitrogen. Add 1ml 2M imidazole, divide the sample into two equal parts, add 1ml 30mg / ml NaBH4 and 1ml 30mg / ml NaBD4 respectively, and react for 3h. Add 100μ2M TFA, react at 121℃ for 90min, and evaporate to dryness at 30℃.

[0087] Add 50μ2M ammonia water and 50μ1M NaBD4, mix well, and react at room temperature for 2.5h. Add 20μ acetic acid to terminate the reaction, blow dry with nitrogen, wash twice with 250 methanol, blow dry with nitrogen. Add 250μ acetic anhydride, vortex mix well, and react at 100℃ for 2.5h. Add 1ml water and let stand for 10min. Add 500μ dichloromethane, vortex mix well, centrifuge, discard the water phase, and repeat washing with water 3 times. Take the lower dichloromethane phase and detect it on GC-MS.

[0088] (2) Instrument parameters

[0089] The analytical instrument used in this embodiment is a 7890A-5977B gas spectrometer / mass spectrometer produced by Agilent Technologies Inc. CA, UAS, and the automatic sampler model is G4567A.

[0090] (3) Chromatographic parameters: The chromatographic system of this embodiment adopts Agile Ent gas chromatography system (Agiile Ent 7890A; Agile Ent Technologies, USA) with chromatographic column: BPX70 (30mx0.25 mmx0.25μm, SGE, Australia). The injection volume is 1μ, the split ratio is 10:1, and the carrier gas is high-purity helium; the initial temperature of the column oven is 140°C for 2.0 min, and the temperature is raised to 230°C at 3°C / min and maintained for 3 min.

[0091] (4) Mass spectrometry parameters: The mass spectrometry system used in this embodiment is a quadrupole mass spectrometry detection system (Agilent 5977B; Agile Technologies, USA) of American Aigent Company, equipped with an electron impact ion source (EI) and a MassHunter workstation. The electron impact ion source (EI) was used, and the analytes were detected in the full scan (SCAN) mode, with a mass scan range (m / z): 50-350.

[0092] (5) Qualitative and quantitative analysis: The principle of gas chromatography is that the sample is vaporized and carried into the chromatographic column by the carrier gas. Since the distribution coefficients of the components in the sample between the carrier gas and the chromatographic column stationary liquid are different, their adsorption or solubility is different, so the running speed and retention time of the components in the chromatographic column are different, and they are separated from each other and enter the detector in a certain order.

[0093] The qualitative principle of mass spectrometry is that after the components to be tested are vaporized and separated by gas chromatography, they are bombarded by ions or electrons in the ion source and lose a valence electron to become a positively charged molecular ion, which is further broken into fragment ions. Under the combined action of the electric field and magnetic field of the mass analyzer, they are separated according to the m / z size, detected and recorded, thereby obtaining characteristic fragments after polysaccharide methylation, and compared with the existing database to confirm its bonding mode.

[0094] The total ion current diagram (D generation) of the sample obtained in this embodiment is as follows Figure 1 As shown, H generation Figure 2 shown.

[0095] This embodiment mainly calculates the relative molar ratio of various glycosidic bonds in the sample by calculating the ratio of the chromatographic peak area to the molecular weight of the corresponding derivative, which approximately represents the relative molar amount of the compound. The sample measurement results are shown in Table 1.

[0096] Table 1 Results of bonding structure analysis of polysaccharide samples

[0097]

[0098] Example 4 NMR analysis

[0099] In this example, the extract obtained in Example 1 was measured and the results were as follows: 1 H-NMR and 13 C-NMR spectrum was used for structural identification and the identification results were obtained.

[0100] (1) Sample pretreatment:

[0101] Take an appropriate amount of purified polysaccharide and fully dissolve it in D20 to prepare a polysaccharide solution with a concentration greater than or equal to 40 mg / mL. Transfer the dissolved solution to an NMR tube and add 0.5 mL. Place the NMR tube in an NMR spectrometer and scan it in one dimension. 1 H spectrum, 13 C spectrum, two-dimensional COSY, HSOC, HMBC, and NOESY spectra.

[0102] (2) Instrument parameters:

[0103] This project uses a Bruker (Germany) 600MHz nuclear magnetic resonance spectrometer to quantitatively analyze the target, with a scanning temperature of 25°C. Liquid probe QXI 1H / 31P / 13C / 15N 5mm quadruple resonance reverse detection probe (Z-gradient, ATMAcc), technical parameters: signal-to-noise ratio (1H): 888: resolution (Hz): 0.32 (rotation) BBFO 1H-19F, 31P-15N, 1H decoupling / observe multi-nuclear forward detection probe (Z-gradient, ATM) technical parameters: signal-to-noise ratio (1H): 798; resolution (Hz): 0.26 (rotation); signal-to-noise ratio (13C): 328; resolution (Hz): 0.1.

[0104] (3) Results:

[0105] 1 H-NMR spectrum Figure 3 One-dimensional hydrogen spectrum mainly solves the configuration problem of glycosidic bonds in polysaccharide structure. 1 The signals in H-NMR are concentrated at 3-6ppm. Usually, the anomeric hydrogen signals of β-glycosidic bond configuration are mainly distributed at 84.3-4.8ppm, and the anomeric hydrogen signals of a-glycosidic bond configuration are mainly distributed at 84.8-5.8ppm. The hydrogen spectrum signals of this sample are mainly concentrated between 83.0 and 5.5ppm. Multiple coupling signal peaks are identified in the δ4.3-5.5ppm anomeric signal area, indicating that this sample contains a variety of sugar residues, and the corresponding chemical shifts of anomeric hydrogen are 65.33, 5.26, 5.0, 4.92, 4.92, 466 and 4.46ppm respectively. The non-anomeric hydrogen signals are mainly concentrated in the 63.2-4.2ppm area. Due to the serious overlap of individual signals, it is necessary to combine COSY and HSOC spectra to attribute the H2-H6 chemical shifts of each sugar residue. Among them, the strong signal peak near 64.71ppm is the solvent peak.

[0106] 13 C-NMR spectrum Figure 4 As shown. 1 Compared with H-NMR, polysaccharides 13 The chemical shift signals in C-NMR are widely distributed. 13 The anomeric carbon signal in C-NMR is concentrated at 95-110 ppm. Multiple signal peaks were identified in the anomeric carbon region of this sample. 13The cross peaks in the anomeric region of the C-NMR spectrum and the HSQC spectrum determined that the anomeric signals present in the sample were: 64.46 / 103.04, 85.26 / 99.79.65.33 / 99.7, 64.92 / 97.93, 64.72 / 102.77, 64.66 / 103.04 and 65.0 / 1015 ppm, which were recorded as sugar residues A, B, C, D, E, F and G, respectively. Combining the bonding structure (methylation) information of the samples, the anomeric signal and the comprehensive reports in the literature, it was inferred that the sugar residue A was →6)-β-DG l cp-(1→, B was →4)-aDG l cp-(1→, C was aDG l cp-(1→, D was →6)-aD-Ga lp-(1→, E was →3)-β-DG l cp-(1→, residue F was →3,6)-β-DG l cp-(1→, G was →4,6)-aD-Ga lp-(1→, and its residues were 1 H and 13 The chemical shifts were assigned and the results are shown in Table 2.

[0107] Hydrogen-hydrogen correlation spectroscopy (COSY) Figure 5 As shown. Hydrogen-hydrogen correlation spectrum (COSY) is the most commonly used homonuclear displacement correlation spectrum. It generally reflects the coupling relationship between adjacent carbon and hydrogen, and thus can know the coupling relationship between protons in the same spin system. It is a powerful tool for assigning spectral lines and deducing structures. 1 The H signals will intersect on the diagonal line, and the intersection is called the diagonal peak. The two points on both sides of the diagonal line on the graph are symmetrically distributed and are called cross peaks (or correlation peaks). 1 The H signals will intersect at the relevant peaks, and the main analysis is of adjacent or interphase proton related signals.

[0108] Hydrogen-hydrogen correlation spectroscopy (NOESY) Figure 6 As shown. NOESY is a two-dimensional nuclear magnetic resonance spectrum of the NOE type. NOE is mainly used to determine whether two protons are close to each other in the three-dimensional structure of a molecule. If NOE exists, it means that the two are close. The larger the NOE value, the closer the distance between the two in space. NOE plays an important role in determining the structure, configuration and conformation of organic compounds.

[0109] Carbon-hydrogen correlation spectroscopy (HSQC) Figure 7 HSQC spectrum is the CH heteronuclear single quantum correlation spectrum. 1 H nucleus and its directly connected 13 C nucleus, the information and spectra provided are related to 13 C- 1The HCOSY spectrum is the same. Like HMBC, it is also measured by high sensitivity 1 H nuclei to detect 13 C- 1 The sensitivity of the long-range coupling information between H is higher than that of the traditional long-range coupling 13 C- 1 H COSY is much higher.

[0110] Carbon-hydrogen correlation spectroscopy (HMBC) Figure 8 As shown. HMBC spectrum is a CH heteronuclear multiple bond correlation spectrum. HMBC can detect the long-range coupling signal of carbon and hydrogen with high sensitivity, that is, the coupling of C and H between two adjacent sugar residues. Through the coupling of anomeric hydrogen (carbon) with the carbon (hydrogen) connected to another sugar residue, the connection fragment and sequence between monosaccharide residues can be inferred.

[0111] Combining the one-dimensional NMR and two-dimensional NMR spectra, the sugar residue signals of the samples were assigned, as shown in Table 2:

[0112] Table 2 Sugar residues 1 H and 13 Chemical shift of C

[0113]

[0114]

[0115] nd: Abbreviation for “not detected”, meaning not identified.

[0116] According to the sugar residues in the sample 13 C and 1H chemical shift, combined with HMBC spectrum analysis of the structure and connection mode in the polysaccharide: sugar residue A-H1 and residue B-C6 have a cross peak of 64.46 / 74.99ppm, and a cross peak of 64.46 / 69.55ppm with residue D-C6. Sugar residue C-H1 and residue G-C4 have a cross peak of 65.33 / 76.86ppm; sugar residue C-C1 and residue G-H4 have a cross peak of 699.7 / 3.59ppm. Sugar residue E-C1 and residue A-H6 have a cross peak of δ102.77 / 3.79ppm. Combined with NOESY spectrum, the connection order of each residue in the polysaccharide was further judged and verified, and sugar residue A-H1 and residue D-H6 had a cross peak of δ4.46 / 3.41ppm. Sugar residue B-H1 has a cross peak with residue A-H6 at δ5.26 / 3.79ppm, and with residue G-H6 at 65.26 / 3.86ppm. Sugar residue C-H1 has a cross peak with residue F-H6 at 65.3313.57ppm. Sugar residue D-H1 has a cross peak with residue A-H6 at 64.92 / 3.79ppm. Sugar residue F-H1 has a cross peak with residue E-H3 at 64.66 / 3.68ppm. Sugar residue G-H1 has a cross peak with residue F-H3 at δ5.0 / 3.74ppm.

[0117] Therefore, based on the analysis of one-dimensional and two-dimensional nuclear magnetic resonance information, it is inferred that the main chain of the polysaccharide is mainly composed of →6)-β-D-Gl cp-(1→4)-aDG l cp-(1→, →6)-aD-Ga lp-(1→ and a small amount of →3)-β-DG l cp-(1→, and the main branch chain is composed of aDG l cp-(1→ connected to the 0-6 position of the sugar residue →3,6)-β-DG l cp-(1→ and the 0-4 position of →4,6)-aDG l cp-(1→. Therefore, the structure of the polysaccharide chain is inferred to be:

[0118]

[0119] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for extracting FA-2-b-β from Qinba selenium mushroom extract, characterized in that: First, the Qinba selenium mushroom raw material is decocted for multiple times to obtain a crude extract, and then the crude extract is subjected to multiple alcohol-water extractions; wherein, the decocting temperature for each time is 80-120°C, and the decocting time for each time is 0.5-2h; the alcohol is an ethanol solution with various volume percentages.

2. The method for extracting FA-2-b-β from the Qinba selenium mushroom extract according to claim 1, characterized in that: The following steps are involved: S1, obtaining the Qinba selenium mushroom raw material, adding water and decocting for 2 to 5 times, filtering and combining the decoctions obtained after each decoction, and then concentrating to obtain the crude extract; S2, adding a first ethanol solution to the crude extract, mixing and filtering to obtain a first precipitate; S3, mixing the first precipitate with water and centrifuging for multiple times, and mixing the supernatants obtained from each centrifugation to obtain a first mixed solution; S4, dialyzing the first mixed solution to obtain a dialysate; S5, adding a second ethanol solution to the dialysate, mixing, standing and filtering to obtain a second precipitate; S6. Wash the second precipitate with a third ethanol solution and then dry it to obtain the Qinba selenium mushroom extract acidic RNA protein complex FA-2-b-β.

3. The method for extracting FA-2-b-β from the Qinba selenium mushroom extract according to claim 2, characterized in that: The volume percentage of ethanol in the first ethanol solution is less than the volume percentage of ethanol in the second ethanol solution, and the volume percentage of ethanol in the second ethanol solution is less than the volume percentage of ethanol in the third ethanol solution.

4. The method for extracting FA-2-b-β from the Qinba selenium mushroom extract according to claim 3, characterized in that: In step S2, the volume percentage of ethanol in the first ethanol solution is 75%.

5. The method for extracting FA-2-b-β from the Qinba selenium mushroom extract according to claim 4, characterized in that: In step S2, the volume ratio of the crude extract to the first ethanol solution is 1:7-9.

6. The method for extracting FA-2-b-β from the Qinba selenium mushroom extract according to claim 3, characterized in that: In step S5, the volume percentage of the second ethanol solution is 90-95%.

7. The method for extracting FA-2-b-β from the Qinba selenium mushroom extract according to claim 6, characterized in that: In step S5, the volume ratio of the dialysate to the second ethanol solution is 1:8-9.

8. The method for extracting FA-2-b-β from the Qinba selenium mushroom extract according to claim 3, characterized in that: In the step S6, the third ethanol solution is anhydrous ethanol.

9. The method for extracting FA-2-b-β from the Qinba Selenium Mushroom extract according to any one of claims 1 to 8, characterized in that: The Qinba selenium mushroom raw material is Qinba selenium mushroom powder, and the mesh number of the Qinba selenium mushroom powder is 80-120 meshes.

10. A Qinba selenium mushroom extract FA-2-b-β, characterized in that The extract is obtained by the method according to any one of claims 1 to 9, wherein the structural formula of the Qinba selenium mushroom extract FA-2-b-β is as shown in formula (1):

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