Four gentlemen soup compound active oligosaccharide and application thereof
Patent Information
- Application Number
- CN202311720036.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-12-14
- Publication Date
- 2026-09-15
- Estimated Expiration
- 2043-12-14
Smart Images
Figure CN117717560B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of natural product development and relates to oligosaccharides, specifically to an active oligosaccharide in the Sijunzi Decoction compound and its use in improving spleen deficiency syndrome. Background Technology
[0002] Oligosaccharides are low-polymerization-degree (DP) carbohydrates composed of monosaccharides linked by glycosidic bonds, with five- or six-carbon sugars as the main building blocks, including glucose, fructose, xylose, and mannose. Studies have shown that oligosaccharides possess various pharmacological and physiological activities, including antidepressant, antioxidant, blood glucose-regulating, immune-regulating, and gut microbiota-regulating effects. Given the abundance and wide range of pharmacological activities of traditional Chinese medicine (TCM) resources, research on TCM oligosaccharides is of great significance and promising prospects, and some progress has already been made. For example, Morinda officinalis oligosaccharides can improve depressive-like behavior in mice by regulating 5-HT levels and HPA axis function; its Morinda officinalis oligosaccharide capsules are now one of the recommended TCM preparations for clinical use in treating depression. Rehmannia glutinosa oligosaccharides can improve glucose metabolism disorders in diabetic and hyperglycemic rats. Arctium lappa oligosaccharides can enhance the immune function of immunocompromised mice and promote the secretion of IL-2 and IFN-γ from the spleen tissue of normal mice. Summary of the Invention
[0003] This invention proposes an active oligosaccharide in the Sijunzi Decoction compound and its application. The active oligosaccharide component SJZD-OGS (SiJunZi Decoction Oligosaccharide) was prepared by multiple separations and purifications from the Sijunzi Decoction compound, a classic formula for clinically treating spleen deficiency. It exhibits significant anti-inflammatory activity and can be used in the preparation of immunomodulatory drugs. In vivo animal experiments have confirmed that SJZD-OGS can improve spleen deficiency syndrome, and can be further developed into a clinical treatment for spleen deficiency. SJZD-OGS can also regulate intestinal flora and its metabolites, short-chain fatty acids (SCFAs), and can be further developed into drugs that regulate the intestinal flora.
[0004] This invention is achieved through the following technical solution:
[0005] This invention relates to an active oligosaccharide component (SJZD-OGS) isolated from the Sijunzi Decoction compound, the effective component of which, by mass percentage, is 44.35% sucrose oligosaccharide, specifically comprising: 5.06% sucrose trisaccharide, 4.83% sucrose pentasaccharide, 6.10% sucrose hexasaccharide, 7.45% sucrose heptasaccharide, 7.33% sucrose heptasaccharide, 6.95% sucrose octasaccharide, and 6.63% sucrose nonasaccharide.
[0006] The SJZD-OGS has a sugar content of 75.0% to 95.0%; a degree of polymerization (DP) of 2 to 14; and a monosaccharide composition including glucose (Glu) and fructose (Fru).
[0007] This invention relates to a method for preparing the above-mentioned active oligosaccharide SJZD-OGS from Sijunzi decoction. The method involves extracting the non-polysaccharide (SJZD-NPSs) component of Sijunzi decoction from the concentrated Sijunzi decoction, eluting it with macroporous resin to obtain the oligosaccharide component APS-1, and then purifying it by activated carbon adsorption filtration.
[0008] The Sijunzi Decoction concentrate mentioned above is produced using, but is not limited to, the methods recorded in the Taiping Huimin Heji Jufang.
[0009] The aforementioned Sijunzi decoction non-polysaccharide (SJZD-NPSs) component is obtained by centrifuging the concentrated Sijunzi decoction solution after alcohol precipitation, then concentrating to remove the solvent, reconstitute with water, evaporate to dryness until the ethanol is completely removed, reconstitute with water, and freeze-dry under reduced pressure.
[0010] The oligosaccharide component APS-1 was obtained by dissolving and loading the non-polysaccharide (SJZD-NPSs) component of Sijunzi decoction into AB-8 macroporous resin, eluting with water, detecting the eluent using the phenol-sulfuric acid method, combining the eluents according to their sugar content, concentrating and drying them to obtain the crude oligosaccharide component APS-1 of Sijunzi decoction.
[0011] The activated carbon adsorption method is as follows: APS-1 is prepared into a 10 mg / mL solution with distilled water. Activated carbon (sample to activated carbon mass ratio 1:4–8) is added and mixed evenly. The mixture is stirred on a magnetic stirrer for 30–60 min, then filtered. The filtrate is the monosaccharide component (water fraction, designated as fraction A). The APS-1-activated carbon mixture is then dissolved, stirred, and filtered sequentially with 3%–8% ethanol, 10%–15% ethanol, and 45%–50% ethanol. The filtrates contain disaccharides (3%–8% ethanol, designated as fraction B; 10%–15% ethanol, designated as fraction C) and oligosaccharides (45%–50% ethanol, designated as fraction D). Fraction D is concentrated and freeze-dried to obtain the Sijunzi decoction oligosaccharide SJZD-OGS active component.
[0012] This invention relates to the application of the oligosaccharide active component SJZD-OGS in Sijunzi Decoction prepared by the above method, which is used to prepare immunomodulatory drugs or health products or drugs to improve spleen deficiency syndrome.
[0013] The aforementioned health products or drugs use Sijunzi decoction oligosaccharide SJZD-OGS as the active component, along with other natural extracts and pharmaceutically acceptable excipients or excipients, to prepare pharmaceutical compositions, including but not limited to tablets, granules, pills, capsules, oral liquids, etc. Attached Figure Description
[0014] Figure 1 This is a schematic diagram of the degree of polymerization analysis of SJZD-OGS;
[0015] Figure 2 This is a schematic diagram of the IC analysis for monosaccharides in SJZD-OGS hydrolysis products;
[0016] Figure 3 The BPI graph for the mixed standard sample;
[0017] Figure 4 BPI plot for SJZD-OGS;
[0018] Figure 5 Secondary fragment diagram of fructotriose;
[0019] Figure 6 Secondary fragment diagram of fructosaccharide;
[0020] Figure 7 A schematic diagram showing the effect of different sites on LPS-induced NO release rate in RAW264.7 cells;
[0021] Note: A: Water fraction; B: 5% ethanol fraction; C: 10% ethanol fraction; D: SJZD-OGS; Compared with the model group, ***P<0.001;
[0022] Figure 8 A schematic diagram illustrating the effect of SJZD-OGS on GAS in rats with spleen deficiency syndrome;
[0023] Figure 9 This is a schematic diagram illustrating the effect of SJZD-OGS on the richness of intestinal flora in rats with spleen deficiency syndrome.
[0024] Figure 10 A schematic diagram illustrating the effect of SJZD-OGS on the diversity of gut microbiota in rats with spleen deficiency syndrome;
[0025] Figure 11 A schematic diagram showing the effect of SJZD-OGS on the total short-chain fatty acid content in rats with spleen deficiency syndrome.
[0026] In the figure: CON: blank control group; SDS: model group; SDS+SJZD: Sijunzi decoction group; SDS+OGS: Sijunzi decoction oligosaccharide group. Compared with the blank group, #P<0.05, ##P<0.01, ###P<0.001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001. Detailed Implementation Example 1
[0027] This embodiment relates to the separation and preparation of oligosaccharide SJZD-OGS based on the traditional Chinese medicine compound Sijunzi Decoction, specifically including:
[0028] Step 1: Take 9g of ginseng (without the root tip), 9g of atractylodes macrocephala, 9g of poria cocos (without the peel, crush them into coarse particles before decoction), and 6g of prepared licorice root. Soak the compound medicinal slices in 10 times the amount of water until thoroughly soaked (soaking time > 2 hours). Then, use a multi-stage electric stove to control the heat for decoction. Each time, bring the water to a boil quickly over high heat, then reduce to low heat and maintain a gentle boil for 30 minutes. Filter the decoction through three layers of gauze while it is still hot. Decoction is repeated twice. Combine the decoctions and concentrate them under reduced pressure at 60℃ to about 33mL. Finally, adjust the concentration of the concentrated decoction with water to 1g of compound medicine / mL to obtain the concentrated Sijunzi Decoction.
[0029] Step 2: Add 4 times the volume of 95% ethanol (1:4, v / v) to the concentrated Sijunzi decoction obtained in Step 1 for alcohol precipitation (slow and fast stirring), place in a refrigerator at 4°C for 48 hours, and centrifuge at 4500 rpm at room temperature for 15 minutes to obtain the supernatant.
[0030] Step 3: Concentrate the supernatant to remove the solvent, add water to reconstitute, evaporate to dryness until the ethanol is completely removed, add water to reconstitute, and freeze-dry under reduced pressure to obtain the non-polysaccharide component of Sijunzi Decoction.
[0031] Step 4: Dissolve the non-polysaccharide component of Sijunzi Decoction in an appropriate amount of water and load it into AB-8 macroporous resin. Elute with water and test the eluent using the phenol-sulfuric acid method. Combine the eluents according to their sugar content, concentrate and dry to obtain the crude oligosaccharide component APS-1 of Sijunzi Decoction.
[0032] Step 5: Prepare a 10 mg / mL solution of 5.67 g APS-1 with distilled water, add 31.02 g activated carbon (sample to activated carbon mass ratio, 1:6) and mix evenly. Stir on a magnetic stirrer for 30 min, filter, and the filtrate is the monosaccharide component (water fraction, named fraction A, yield 39.08%).
[0033] Step 6: Dissolve, stir and filter the APS-1 activated carbon mixture sequentially with 5% ethanol, 10% ethanol and 50% ethanol. The filtrates contain disaccharides (5% ethanol, named part B, yield 16.28%; 10% ethanol, named part C, yield 8.24%) and oligosaccharides (50% ethanol, named part D, yield 5.37%).
[0034] Step 7: Concentrate and freeze-dry the above-mentioned D fraction to obtain Sijunzi decoction oligosaccharide SJZD-OGS. Table 1 shows the yield of each fraction and the sample recovery rate.
[0035] Table 1 Summary of sample yield and recovery rate from four locations
[0036] The active oligosaccharide component SJZD-OGS of the prepared compound Sijunzi decoction was subjected to component analysis and structural characterization, specifically including:
[0037] Total sugar content analysis: The sugar content of SJZD-OGS was determined using the phenol-sulfuric acid method. First, D-(+)-glucose reference standard (pre-dried in a 50℃ oven for 1 h) was accurately weighed and prepared into a standard solution with a concentration of 100 μg / mL. Accurately pipette 0, 0.2, 0.4, 0.6, 1.2, and 1.6 mL of the standard solution into 20 mL stoppered glass tubes, add distilled water to a final volume of 2.0 mL, add 1 mL of 5% phenol solution and shake well. Finally, add 5 mL of concentrated sulfuric acid and mix well. Heat in a boiling water bath for 20 min, remove and cool to room temperature, and measure the absorbance at 490 nm using ultraviolet spectrophotometry. Accurately weigh the sample and prepare a sample solution with a concentration of 50 μg / mL. Accurately pipette 2.0 mL of the sample solution into a stoppered test tube, repeat the above steps, and measure the absorbance of the sample. The results showed that the total sugar content of SJZD-OGS was 89.72%.
[0038] Degree of polymerization analysis of SJZD-OGS by MALDI-TOF / TOF MS: SJZD-OGS and 2,5-dihydroxybenzoic acid (DHB) matrix were mixed at a 1:1 ratio on a MALDI plate. The degree of polymerization of SJZD-OGS was analyzed by matrix-assisted laser desorption / ionization tandem time-of-flight mass spectrometry (MALDI-TOF / TOF 7090). The instrument used MALDI-7090 ionization (Tuning Linear, Power 73, P.Extat 3000.00 (bin 189), Lon Gate Blanking: 500.00, Laser Diameter: 200) to introduce ions generated by laser desorption / ionization into the TOF analyzer, and mass spectrometry was performed in the m / z range of 100-5000. The results are as follows: Figure 1 As shown, the degree of polymerization (DP) of SJZD-OGS is 3-14.
[0039] Ion chromatography (IC) analysis of the monosaccharide composition of SJZD-OGS: First, 3 mg of SJZD-OGS was accurately weighed and dissolved in 1 mL of trifluoroacetic acid (TFA, 2 mol / L). The mixture was reacted at 100 °C for 6 h to achieve complete hydrolysis, yielding the hydrolysis product. After drying under nitrogen, methanol was added three times to remove residual TFA. The final sample concentration was then dissolved in water to achieve a final concentration of 300 μg / mL. Ion chromatography was used to analyze the monosaccharide components in the SJZD-OGS hydrolysis product. Fucose (Fuc), rhamnose (Rha), galactosamine hydrochloride (GalN), arabinose (Ara), glucosamine (GlcN), galactose (Gal), glucose (Glu), mannose (Man), xylose (Xyl), fructose (Fru), and glucuronic acid (GluA) were accurately weighed and prepared into mixed standard solutions of different concentrations using ddH2O. After centrifugation, the supernatant was transferred to a liquid chromatography vial for later use. The chromatographic conditions were as follows: separation was performed using an ICS-5000+ ion chromatography system (Thermofisher, USA) with a Thermo CarboPac PA100 column (9 mm × 250 mm, Thermofisher). Gradient elution was performed using 50 mM sodium hydroxide solution (A), 1 M sodium acetate solution (B), 400 mM sodium hydroxide solution (C), and aqueous solution (D) as eluents. The column temperature was 30 °C, the flow rate was 0.5 mL / min, and the detection wavelength was 250 nm, detected by an amperometric detector. Results are as follows: Figure 2 As shown in Table 2, the hydrolysis products of SJZD-OGS are mainly composed of glucose (Glu) and fructose (Fru), with a molar ratio of Glu:Fru = 1.00:0.0039.
[0040] Table 2 Summary of Monosaccharide Composition Analysis of SJZD-OGS Hydrolysis Products
[0041] Qualitative Analysis of SJZD-OGS by UPLC-IMS-QTOF / MS: Related studies have shown that Sijunzi Decoction may contain oligosaccharide components of inulin-type fructans. Therefore, we performed qualitative analysis of SJZD-OGS by UPLC-IMS-QTOF / MS in conjunction with sucrose oligosaccharide standards. First, accurately weighed SJZD-OGS and prepared a sample solution with acetonitrile-water (1:1, v / v) to a concentration of 2 mg / mL. The solution was centrifuged at 12000 rpm at room temperature for 10 min, and the supernatant was transferred to a liquid chromatography vial for later use. Accurately weighed sucrose trisaccharides to sucrose nonaconitoses and prepared a mixed standard solution with acetonitrile-water (1:1, v / v) to a concentration of 100 μg / mL. The solution was centrifuged at 12000 rpm at room temperature for 10 min, and the supernatant was transferred to a liquid chromatography vial for later use. Chromatographic conditions were as follows: Separation was performed using an Acquity UPLC I-class (Waters Ltd., USA) column, with a Waters XBridge HILIC (2.1 × 100 mm, Waters) and a flow rate of 0.4 mL / min. The mobile phase consisted of gradient elution with different ratios of 0.1% formic acid-water and 0.1% formic acid-acetonitrile, with an injection volume of 5 μL and a column temperature of 40 °C. Mass spectrometry detection conditions were as follows: Data acquisition was performed using a VION IMS QTOF mass spectrometer (Waters Ltd., USA) in negative ion (ESI-) mode, with an m / z acquisition range of 100-2000. The baseline chromatogram and BPI chromatogram of the oligosaccharide mixed standard are shown below. Figure 3 The BPI chart of SJZD-OGS is as follows: Figure 4 The results showed that the retention times of the ion peaks in SJZD-OGS were consistent with those of the oligosaccharide standard. Further comparison of ion fragment information and literature review (Table 3) identified disaccharides, fructotrioses, and fructononoses in SJZD-OGS. Taking fructotriose (1-Ketose) as an example (secondary fragment information is shown in Table 3)... Figure 5 Fragment ions were detected at m / z 503.1614, 341.1093, 191.0538, 173.0452, and 161.0453, respectively, in negative ion mode. Among these, m / z 503.1614 represents the molecular ion peak [MH] of 1-Ketose. - The ion fragment produced at m / z 341.1093 is [MH-C6H]. 10 O5] - When the molecular ion peak loses two C6H atoms... 10 After O5 and an H2O molecule, at m / z 161.0453 [MH-2C6H 10 O5-H2O] - Fragment ions are formed at the site. Simultaneously, secondary fragments of nystose (secondary fragment information is as follows) are formed. Figure 6 It contains fragment ions at m / z 665.2123, 503.1614, 341.1070, 191.0555, and 179.0563, with m / z 665.2123 being the molecular ion peak of Nystose [MH]. - When the molecular ion peak loses a C6H... 10 Following the O5 molecule, with m / z 503.1614 [MH-C6H] 10 O5] - Fragment ions are formed at the site; two C6H atoms are lost. 10 Following the O5 molecule, with m / z 341.1070 [MH-2C6H] 10 O5] - Fragment ions are formed at the site; three C6H atoms are lost. 10 Following the O5 molecule, with m / z 179.0563 [MH-3C6H] 10 O5] - Fragment ions are formed at the site. These results indicate that sucrose oligosaccharides may be one of the main oligosaccharide components in SJZD-OGS.
[0042] Table 3. Information on oligosaccharide components and ion fragments.
[0043] Quantitative Analysis of SJZD-OGS by HPLC-ELSD: Based on the qualitative analysis results of SJZD-OGS above, the oligosaccharide components in SJZD-OGS were further quantitatively analyzed by HPLC-ELSD. Accurately weighed SJZD-OGS was prepared into a sample solution with a concentration of 2 mg / mL using acetonitrile-water (1:1, v / v). The solution was centrifuged at 12000 rpm for 10 min at room temperature, and the supernatant was transferred to a liquid chromatography vial for later use. Accurately weighed fructotriose to fructononose was prepared into mixed standard solutions of different concentrations using acetonitrile-water (1:1, v / v). These solutions were centrifuged at 12000 rpm for 10 min at room temperature, and the supernatant was transferred to a liquid chromatography vial for later use. Chromatographic conditions were as follows: An ALLIANCE e2695 (Waters Ltd., USA) column was used for separation. An XBridge BEH Amide Column (3.5 μm, 4.6 × 150 nm, Waters) was selected, with a flow rate of 0.5 mL / min. The mobile phase consisted of gradient elution with water and acetonitrile in different proportions. The injection volume was 10 μL, and the column temperature was 40 °C. An evaporative light scattering (ELSD, Waters) detector was used, with a nitrogen pressure of 2.5 Bar and a gain of 7. The linear correlation coefficient R0 for the seven sucrose oligosaccharides was calculated. 2All values were greater than 0.99, indicating that the oligosaccharide standard had a good linear relationship, with a linear range between 12.5 and 800.0 μg / mL, as shown in Figure 4. The quantitative analysis results of the seven sucrose oligosaccharides in SJZD-OGS are shown in Table 5. The total content of the seven sucrose oligosaccharides in SJZD-OGS accounted for 44.35%.
[0044] Table 4. Summary of regression equations, linear ranges, and contents of sucrose trisaccharides to nonasaccharides.
[0045] Table 5 Quantitative analysis results of SJZD-OGS
[0046] In summary, the specific components and their mass percentages of the SJZD-OGS oligosaccharide prepared in this embodiment are as follows: 5.06% fructotriose, 4.83% sucrose, 6.10% fructopentose, 7.45% fructohexaose, 7.33% fructoheptaose, 6.95% fructooctaose, and 6.63% fructononose. Example 2
[0047] This embodiment relates to the application of the oligosaccharide SJZD-OGS prepared based on the above method, specifically for the preparation of immunomodulatory drugs. This includes establishing an in vitro LPS-induced mouse mononuclear macrophage leukemia RAW 264.7 cell inflammation model and observing its anti-inflammatory effect. Specifically, RAW 264.7 cells were cultured in DMEM medium (containing 10% fetal bovine serum and 1% penicillin / streptomycin) and incubated at 37°C under constant temperature and humidity. RAW 264.7 cells in the logarithmic growth phase were harvested and cultured at a cell density of 3 × 10⁻⁶ cells. 5 Cells were seeded at a concentration of 1 / mL in 96-well plates and cultured for 24 h. The four fractions A, B, C, and D (SJZD-OGS) isolated by activated carbon adsorption in Example 1 were prepared into drug delivery solutions of 50, 100, 200, and 400 μg / mL in DMEM medium, respectively. SJZD decoction was used as a positive control. RAW 264.7 cells were incubated for 1 h, followed by induction with 1 μg / mL LPS. After 24 h, the supernatant was analyzed using a Beyotime NO detection kit. Experimental data are expressed as mean ± sem. T-tests were used to compare pairwise differences between groups; a p-value less than 0.05 was considered statistically significant.
[0048] The results are as follows Figure 7As shown, compared with the blank group, the NO release rate in the model group was significantly increased, indicating that the RAW 264.7 cell inflammation model was successfully established. Compared with the model group, the anti-inflammatory activity of the four sites A, B, C, and D was concentrated in site D, namely SJZD-OGS, which was able to reduce the LPS-induced NO release level in cells in a dose-dependent manner, and the effect was better than that of the positive control SJZD group. The above results suggest that SJZD-OGS has good immunomodulatory activity in vitro. Example 3
[0049] This embodiment relates to the application of SJZD-OGS, an oligosaccharide prepared based on the above method, specifically for the preparation of a drug to improve spleen deficiency syndrome. The specific method involves establishing a rat model of spleen deficiency syndrome using a multi-factor modeling approach, and then administering SJZD-OGS along with the SJZD-OGS compound to improve the spleen deficiency syndrome. The details are as follows: Male SD rats, 180-200g, were randomly divided into four groups after one week of acclimatization: ① Blank control group (CON); ② Spleen deficiency model group (SDS); ③ Spleen deficiency + SJZD-OGS decoction group (SDS+SJZD); ④ Spleen deficiency + SJZD-OGS group (SDS+OGS). The modeling method was as follows: Rhubarb aqueous extract was administered by gavage daily. From day 1 to day 7, the gavage dose of rhubarb was 10 g / kg, and from day 8 to day 21, the gavage dose was 12 g / kg. Simultaneously, rats were subjected to a starvation-starvation model, consisting of one day of full feeding followed by two days of starvation. On the first day of starvation, rats swam to exhaustion (water temperature approximately 20℃; the criterion for assessment was that the rat's nose was submerged for 5 seconds, and its body sank and could not float). This was repeated for 3 days as one cycle, and rat feces were collected every 3 days. During the modeling period, rats in the blank control group were simultaneously administered distilled water by gavage. After the modeling was completed on day 21, rats were treated with SJZD and SJZD-OGS for 7 days. The dosage of SJZD was 10 g / kg, and the dosage of SJZD-OGS was 250 mg / kg. Samples were collected after the treatment period. The gastrin (GAS) level, a gastrointestinal hormone-related factor, was detected using an ELISA kit; the abundance and diversity of gut microbiota were analyzed by 16S rRNA sequencing; and the total short-chain fatty acid (SCFA) content was detected by GC-MS. Experimental data are expressed as mean ± sem. Pairwise differences between groups were compared using t-tests, with a p-value less than 0.05 considered statistically significant. The effect of SJZD-OGS on GAS levels in rats with spleen deficiency syndrome is as follows: Figure 8 Compared with the control group, the gastrin level in rats with spleen deficiency syndrome showed an increasing trend. Both SJZD and SJZD-OGS administration reduced the expression level of GAS in rats with spleen deficiency syndrome, with SJZD showing a stronger effect than SJZD-OGS. The effect of SJZD-OGS on the richness of intestinal flora in rats with spleen deficiency syndrome is as follows: Figure 9Compared with the control group, the ACE index of rats with spleen deficiency syndrome was significantly decreased. SJZD-OGS administration significantly increased the ACE index of rats with spleen deficiency syndrome, and the effect of SJZD-OGS was stronger than that of SJZD. The effect of SJZD-OGS on the gut microbiota diversity of rats with spleen deficiency syndrome is as follows: Figure 10 Compared with the control group, the Shannon index of rats with spleen deficiency syndrome was significantly reduced. Both SJZD and SJZD-OGS significantly upregulated the Shannon index in rats with spleen deficiency syndrome, with SJZD-OGS showing a stronger effect than SJZD. The effect of SJZD-OGS on the total amount of short-chain fatty acids (SCFAs) in rats with spleen deficiency syndrome is as follows: Figure 11 Compared with the blank group, the total SCFAs content of rats with spleen deficiency syndrome was significantly reduced. Both SJZD and SJZD-OGS administration significantly upregulated the total SCFAs content of rats with spleen deficiency syndrome, and SJZD was more effective than SJZD-OGS.
[0050] The above-described specific implementations can be partially adjusted by those skilled in the art in different ways without departing from the principles and purpose of the present invention. The scope of protection of the present invention is defined by the claims and is not limited to the above-described specific implementations. All implementation schemes within the scope of the claims are bound by the present invention.
Claims
1. A type of oligosaccharide SJZD-OGS for Sijunzi decoction, characterized in that, Its active ingredient, namely sucrose oligosaccharides, has a mass percentage of 44.35%, specifically including: sucrose trisaccharide 5.06%, sucrose 4.83%, sucrose pentasaccharide 6.10%, sucrose hexasaccharide 7.45%, sucrose heptasaccharide 7.33%, sucrose octasaccharide 6.95%, and sucrose nonasaccharide 6.63%. The aforementioned Sijunzi decoction oligosaccharide SJZD-OGS is obtained by extracting the Sijunzi decoction non-polysaccharide (SJZD-NPSs) component from the concentrated Sijunzi decoction, eluting it with macroporous resin to obtain the oligosaccharide component APS-1, and then purifying it by activated carbon adsorption filtration. Specifically, APS-1 is prepared into a 10 mg / mL solution with distilled water, activated carbon is added and mixed evenly, and the mixture is stirred on a magnetic stirrer for 30-60 min. After filtration, the filtrate is the monosaccharide component. Then, the APS-1-activated carbon mixture is dissolved, stirred and filtered successively with 3%-8% ethanol, 10%-15% ethanol and 45%-50% ethanol. The filtrates contain disaccharides and oligosaccharide components containing 45%-50% ethanol, respectively. The oligosaccharide components are concentrated and freeze-dried to obtain Sijunzi decoction oligosaccharide SJZD-OGS.
2. The oligosaccharide SJZD-OGS of Sijunzi decoction according to claim 1, characterized in that, The oligosaccharide active component has a sugar content of 75.0% to 95.0%; a degree of polymerization (DP) of 2 to 14; and a monosaccharide composition including glucose (Glu) and fructose (Fru).