Process method for preparing D-chiro-inositol by taking buckwheat as raw material
By combining ultrasound-assisted ethanol extraction, Saccharomyces cerevisiae fermentation, and α-galactosidase hydrolysis with resin column and membrane filtration technologies, the problem of low yield and purity of D-chiral inositol prepared from buckwheat has been solved, achieving efficient and environmentally friendly preparation of high-purity D-chiral inositol.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- ZHUCHENG HAOTIAN PHARMA CO LTD
- Filing Date
- 2026-01-27
- Publication Date
- 2026-05-15
AI Technical Summary
Existing technologies for preparing D-chiral inositol from buckwheat have problems such as low yield and low purity, and there are safety risks associated with genetically modified starch conversion and halogenated benzene chemical synthesis methods.
Ultrasonic-assisted ethanol extraction was used to disrupt cell structure, followed by resin column treatment to remove pigments and impurities. D-chiral inositol was converted by fermentation with Saccharomyces cerevisiae and enzymatic hydrolysis with α-galactosidase. Clarification and fractionation were performed by combining ceramic membranes and ultrafiltration membranes. Finally, high-purity products were obtained by desalting with anion and cation exchange resins and crystallizing with ethanol precipitation.
It achieves high-yield, high-purity D-chiral inositol preparation with a green and environmentally friendly process suitable for large-scale production.
Smart Images
Figure CN122038487A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of D-chiral inositol technology, and more particularly to a process for preparing D-chiral inositol from buckwheat. Background Technology
[0002] Currently, common methods for obtaining D-chiral inositol include: natural plant extraction, artificial synthesis via genetically modified starch conversion, and chemical synthesis via halogenated benzene. Among these, the artificial synthesis via genetically modified starch conversion and the chemical synthesis via halogenated benzene use raw materials and solvents that pose safety risks, potentially introducing substances harmful to human health. Natural plant extraction commonly uses buckwheat as a raw material, but this method suffers from low yield and low purity. Therefore, to address these technical issues, it is necessary to develop a process for preparing D-chiral inositol using buckwheat as a raw material. Summary of the Invention
[0003] The technical problem to be solved by the present invention is to provide a process for preparing D-chiral inositol from buckwheat as raw material, which can achieve high yield, high purity and high output of D-chiral inositol product, in order to address the shortcomings of the prior art.
[0004] To solve the above-mentioned technical problems, the technical solution of the present invention is as follows:
[0005] A process for preparing D-chiral inositol from buckwheat, the process comprising the following steps:
[0006] (1) Take the buckwheat grains after removing impurities, crush and sieve them, add ethanol to the collected buckwheat flour, heat and ultrasonically extract, filter, and use the collected filtrate for later use.
[0007] (2) Take the filtrate from step (1), concentrate it, dilute the collected concentrate with purified water, and put the resulting liquid into the resin column. Collect the effluent for later use.
[0008] (3) Take the effluent from step (2), concentrate it, adjust the pH of the collected concentrate to acidic, add brewing yeast for fermentation, sterilize and cool it after fermentation, and use the fermentation liquid for later use.
[0009] (4) Take the fermentation liquid described in step (3), add purified water to dilute it, and then add α-galactosidase to perform enzymatic hydrolysis. The resulting enzymatic hydrolysate is then ready for use.
[0010] (5) Take the enzymatic hydrolysate described in step (4), filter it through a ceramic membrane, collect the first filtrate, filter it through an ultrafiltration membrane, collect the second filtrate, first enter the cation exchange resin column, and then enter the anion exchange resin column to obtain the second filtrate for later use.
[0011] (6) Take the second effluent from step (5), concentrate it through nanofiltration membrane to obtain the first concentrate, then concentrate it under vacuum, add ethanol to the second concentrate, cool, filter and dry to obtain D-chiral inositol product.
[0012] As an improved technical solution, in step (1), the buckwheat flour and ethanol are added at a ratio of 1:10-16, and the volume concentration of the ethanol is 30-70%.
[0013] As an improved technical solution, in step (1), the temperature is heated to 30-60℃, the ultrasonic power is 300-500W, and the ultrasonic treatment time is 1-2h.
[0014] As an improved technical solution, in step (2), the filtrate is concentrated until there is no alcohol odor, and purified water of 1-2 times the volume of the concentrated solution is added to the obtained concentrated solution; the feed solution enters the resin column at a flow rate of 1-2 BV / h.
[0015] As an improved technical solution, the solid content of the concentrate in step (3) is 25-35wt%, and the pH is adjusted to 5.0-5.5 using glacial acetic acid; the brewing yeast and the concentrate are added at a dosage of 0.3-0.7mg / ml, and the activity of the brewing yeast is 10^8-10^11 CFU / g; the fermentation temperature is 30-37℃, and the fermentation time is 2-3 days.
[0016] As an improved technical solution, in step (4), 2-3 times the volume of purified water is added to the fermentation liquid, the enzyme activity of the α-galactosidase is 5000-8000U / g, and the α-galactosidase is added at a dosage of 1-2g / ml.
[0017] As an improved technical solution, the temperature of the enzymatic hydrolysis treatment in step (4) is 40-45℃ and the time of the enzymatic hydrolysis treatment is 5-8h.
[0018] As an improved technical solution, the pore size of the ceramic membrane in step (5) is 150-300nm, and the molecular weight cutoff of the ultrafiltration membrane is 5000-10000Da; the second filtrate enters the cation exchange resin column at a flow rate of 1-2BV / h, and the first effluent enters the anion exchange resin column at a flow rate of 0.5-1.0BV / h.
[0019] As an improved technical solution, the molecular weight cutoff of the nanofiltration membrane in step (6) is 150-300 Da; the solid content of the second concentrate is 40-50 wt%, and 0.7-0.8 times the volume of the second concentrate of ethanol is added, and the temperature is lowered to 20-25℃.
[0020] After adopting the above technical solution, the beneficial effects of the present invention are:
[0021] In the process of this invention, step (1) uses ultrasound-assisted ethanol extraction to effectively destroy cell structure and improve the dissolution efficiency of target components; in step (2), resin column treatment can initially remove pigments and some flavonoids, reducing the burden of subsequent purification; in step (3), fermentation with Saccharomyces cerevisiae and enzymatic hydrolysis with α-galactosidase can promote the conversion of D-chiral inositol in the form of galactosyl derivatives into free D-chiral inositol, which is dissolved in the solvent. Moreover, Saccharomyces cerevisiae cells have high sugar tolerance and can grow and ferment in a high sugar concentration environment to produce ethanol and carbon dioxide, which greatly improves the extraction rate of free D-chiral inositol. In step (4), the ceramic membrane and ultrafiltration membrane are used together to achieve efficient clarification and fractionation, ensuring the stable operation of subsequent resin and nanofiltration systems; the combined desalting and impurity removal of anion and cation resins significantly improves the purity of the product; in step (5), nanofiltration concentration not only achieves selective enrichment of low molecular weight substances, but also effectively removes inorganic salts and small molecule impurities; finally, combined with ethanol precipitation and low-temperature crystallization, a high-purity, highly crystalline chiral inositol product is obtained. The entire process is green and environmentally friendly, with mild conditions, and is suitable for large-scale production, enabling the efficient and sustainable extraction of high-value-added chiral inositol from natural plant raw materials. Attached Figure Description
[0022] Figure 1 The image shows the liquid chromatography chromatogram of D-chiral inositol in Example 2. Detailed Implementation
[0023] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.
[0024] Example 1
[0025] A process for preparing D-chiral inositol from buckwheat includes the following steps:
[0026] (1) Take the buckwheat grains after removing impurities, crush them, pass them through a 60-mesh sieve and collect 100 kg of buckwheat flour. Add 30% ethanol at a ratio of 1:10, heat to 30°C, ultrasonically extract at 300 W power for 1 h, filter, and collect 1000 L of filtrate for later use.
[0027] (2) Take 1000L of the filtrate from step (1), concentrate it under vacuum until there is no alcohol smell, collect 700L of the concentrate, dilute it with purified water equal to the volume of the concentrate, and enter the resulting liquid into a 700L resin column (filler is D101-resin) at a flow rate of 1BV / h, and then use 600L of purified water to top the liquid. The collected 2000L of effluent is reserved for later use.
[0028] (3) Take 2000L of the effluent from step (2), and after vacuum concentration, obtain 400L of concentrate with a solid content of 25wt%. Adjust the pH to 5.0 with glacial acetic acid, and add 120g of brewing yeast (10^8-10^11 CFU / g) at a dosage of 0.3mg / ml for fermentation (30℃, fermentation time is 2 days). After fermentation, sterilize at 90℃ for 30min and cool to room temperature to obtain 450L of fermentation liquid for later use.
[0029] (4) Take 450L of fermentation liquid from step (3), add twice the volume of purified water to dilute it, and then add 450g of α-galactosidase (enzyme activity of 5000-8000U / g) at a dosage of 1g / L. After enzymatic hydrolysis (enzymatic hydrolysis temperature of 40℃, enzymatic hydrolysis time of 5h), 1400L of enzymatic hydrolysate (containing the water washing equipment) is obtained for later use.
[0030] (5) Take 1400L of the enzymatic hydrolysate from step (4), filter it through a ceramic membrane (pore size of 150-300nm), and then filter the first filtrate collected through an ultrafiltration membrane (molecular weight cutoff of 5000-10000Da). The collected 1600L of the second filtrate is first fed into an 800L cation exchange resin column (packing material is macroporous strong acid cation exchange resin-D001) at a flow rate of 1BV / h, and then fed with 1300L of purified water. The resulting 2900L of the first effluent is then fed into a 1000L anion exchange resin column (packing material is macroporous weak basic anion exchange resin-D201) at a flow rate of 0.5BV / h, and then fed with 1100L of purified water. The resulting 4000L of the second effluent is reserved for use.
[0031] (6) Take 4000L of the second effluent from step (5), concentrate it through a nanofiltration membrane (with a molecular weight cutoff of 150-300Da) to obtain the first concentrate, and then concentrate it under vacuum to obtain 200L of the second concentrate (with a solid content of 40wt%). Add 0.7 times the volume of the second concentrate to the second concentrate, cool it to 20°C, filter and dry it to obtain the D-chiral inositol product.
[0032] Example 2
[0033] A process for preparing D-chiral inositol from buckwheat includes the following steps:
[0034] (1) Take the buckwheat grains after removing impurities, crush them, pass them through a 60-mesh sieve and collect 100 kg of buckwheat flour. Add 50% ethanol at a ratio of 1:13, heat to 50°C, and ultrasonically extract at 420 W for 1.5 h. Filter and collect 1300 L of filtrate for later use.
[0035] (2) Take 1300L of the filtrate from step (1), concentrate it under vacuum until there is no alcohol smell, collect 600L of the concentrate, dilute it with 1.5 times the volume of the concentrate with purified water, and enter the resulting liquid into a 700L resin column (filled with D101-resin) at a flow rate of 1.5BV / h, and then use 1000L of purified water to top the liquid. The collected 2500L of effluent is reserved for later use.
[0036] (3) Take 2500L of the effluent from step (2), and after vacuum concentration, obtain 350L of concentrate with a solid content of 30wt%. Adjust the pH to 5.3 with glacial acetic acid, and add 175g of brewing yeast (10^8-10^11 CFU / g) at a dosage of 0.5mg / ml for fermentation (35℃, fermentation time is 2.5 days). After fermentation, sterilize at 93℃ for 35min and cool to room temperature to obtain 400L of fermentation liquid for later use.
[0037] (4) Take 400L of fermentation liquid from step (3), add 2.5 times the volume of purified water to dilute it, and then add 600g of α-galactosidase (enzyme activity of 5000-8000U / g) at a dosage of 1.5g / L. After enzymatic hydrolysis (enzymatic hydrolysis temperature of 43℃, enzymatic hydrolysis time of 7h), 1450L of enzymatic hydrolysate (containing the water washing equipment) is obtained for later use.
[0038] (5) Take 1450L of the enzymatic hydrolysate from step (4), filter it through a ceramic membrane (pore size of 150-300nm), and then filter the first filtrate collected through an ultrafiltration membrane (molecular weight cutoff of 5000-10000Da). The collected 2050L of the second filtrate is first fed into an 800L cation exchange resin column (packing material is macroporous strong acid cation exchange resin-D001) at a flow rate of 1.5BV / h, and then fed with 1100L of purified water. The resulting 3150L of the first effluent is then fed into a 1000L anion exchange resin column (packing material is macroporous weak basic anion exchange resin-D201) at a flow rate of 0.8BV / h, and then fed with 1500L of purified water. The resulting 4650L of the second effluent is ready for use.
[0039] (6) Take 4650L of the second effluent from step (5), concentrate it through a nanofiltration membrane (with a molecular weight cutoff of 150-300Da) to obtain the first concentrate, and then concentrate it under vacuum to obtain 230L of the second concentrate (with a solid content of 45wt%). Add 0.75 times the volume of the second concentrate to the second concentrate, cool it to 23°C, filter and dry it to obtain the D-chiral inositol product.
[0040] Example 3
[0041] A process for preparing D-chiral inositol from buckwheat includes the following steps:
[0042] (1) Take the buckwheat grains after removing impurities, crush them, pass them through a 60-mesh sieve and collect 100 kg of buckwheat flour. Add 70% ethanol at a ratio of 1:16, heat to 60°C, ultrasonically extract at 500 W for 2 h, filter, and collect 1600 L of filtrate for later use.
[0043] (2) Take 1600L of the filtrate from step (1), vacuum concentrate it until there is no alcohol smell, collect 500L of the concentrate, dilute it with purified water twice the volume of the concentrate, and enter the resulting liquid into a 700L resin column (filled with D101-resin) at a flow rate of 2BV / h, and then use 500L of purified water to top the liquid. The collected 2000L of effluent is reserved for later use.
[0044] (3) Take 2000L of the effluent from step (2), and after vacuum concentration, obtain 300L of concentrate with a solid content of 35wt%. Adjust the pH to 5.5 with glacial acetic acid, and add 220g of brewing yeast (10^8-10^11 CFU / g) at a dosage of 0.7mg / ml for fermentation (37℃, fermentation time is 3 days). After fermentation, sterilize at 95℃ for 40min and cool to room temperature to obtain 400L of fermentation liquid for later use.
[0045] (4) Take 400L of fermentation liquid from step (3), add 3 times the volume of purified water to dilute it, and then add 800g of α-galactosidase (enzyme activity of 5000-8000U / g) at a dosage of 2g / L. After enzymatic hydrolysis (enzymatic hydrolysis temperature of 45℃, enzymatic hydrolysis time of 8h), 1600L of enzymatic hydrolysate is obtained for later use.
[0046] (5) Take 1600L of the enzymatic hydrolysate from step (4), filter it through a ceramic membrane (pore size of 150-300nm), and then filter the first filtrate collected through an ultrafiltration membrane (molecular weight cutoff of 5000-10000Da). The collected 2200L of the second filtrate is first fed into an 800L cation exchange resin column (packing material is macroporous strong acid cation exchange resin-D001) at a flow rate of 2BV / h, and then fed with 1000L of purified water. The resulting 3200L of the first effluent is then fed into an 1000L anion exchange resin column (packing material is macroporous weak basic anion exchange resin-D201) at a flow rate of 1.0BV / h, and then fed with 1400L of purified water. The resulting 4600L of the second effluent is ready for use.
[0047] (6) Take 4600L of the second effluent from step (5), and concentrate it through a nanofiltration membrane (with a molecular weight cutoff of 150-300Da) to obtain the first concentrate. Then concentrate it under vacuum to obtain 220L of the second concentrate (with a solid content of 50wt%). Add 0.8 times the volume of the second concentrate to the second concentrate, cool it to 25°C, filter and dry it to obtain the D-chiral inositol product.
[0048] To better demonstrate that the process of the present invention can yield high-yield, high-purity D-chiral inositol products, the following comparative examples are given with reference to Example 2. The D-chiral inositol products obtained in Examples 1-3 and the comparative examples are detailed in Table 1.
[0049] Comparative Example 1
[0050] Unlike Example 2, ultrasonic extraction was not used in step 1, but the rest of the operations were the same.
[0051] Comparative Example 2
[0052] Unlike Example 2, steps (3) and (4) are missing, and the effluent collected in step (2) is processed in steps (5) and (6).
[0053] Comparative Example 3
[0054] Unlike Example 2, the pH was adjusted to 4.5 in step (3), while the rest of the operation was the same.
[0055] Comparative Example 4
[0056] Unlike Example 2, the pH is adjusted to 6 in step (3), while the rest of the operations are the same.
[0057] Comparative Example 5
[0058] Unlike Example 2, the fermentation temperature in step (3) is 28°C, while the rest of the operation is the same.
[0059] Comparative Example 6
[0060] Unlike Example 2, the fermentation temperature in step (3) is 40°C, while the rest of the operation is the same.
[0061] Comparative Example 7
[0062] Unlike Example 2, the temperature of the enzymatic hydrolysis in step (4) is 35°C, while the rest of the operation is the same.
[0063] Comparative Example 8
[0064] Unlike Example 2, the temperature of the enzymatic hydrolysis in step (4) is 50°C, while the rest of the operation is the same.
[0065] Comparative Example 9
[0066] Unlike Example 2, the enzymatic hydrolysis time in step (4) is 4 hours, while the rest of the operation is the same.
[0067] Comparative Example 10
[0068] Unlike Example 2, the enzymatic hydrolysis time in step (4) is 9 hours, while the rest of the operation is the same.
[0069] Comparative Example 11
[0070] Unlike Example 2, the ultrasonic power in step (1) is 200W, while the rest of the operation is the same.
[0071] Comparative Example 12
[0072] Unlike Example 2, the ultrasonic power in step (1) is 600W, while the rest of the operation is the same.
[0073]
[0074] The data in Table 1 show that the D-chiral inositol prepared by the process method of Example 2 of the present invention has better overall yield, purity and efficiency than other examples and comparative examples.
[0075] 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, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.
Claims
1. A process for preparing D-chiral inositol from buckwheat, characterized in that, The process includes the following steps: (1) Take the buckwheat grains after removing impurities, crush and sieve them, add ethanol to the collected buckwheat flour, heat and ultrasonically extract, filter, and collect the filtrate for later use. (2) Take the filtrate from step (1), concentrate it, dilute the collected concentrate with purified water, and put the resulting liquid into the resin column. Collect the effluent for later use. (3) Take the effluent from step (2), concentrate it, adjust the pH of the collected concentrate to acidic, add brewing yeast for fermentation, sterilize and cool it after fermentation, and use the fermentation liquid for later use. (4) Take the fermentation liquid described in step (3), add purified water to dilute it, and then add α-galactosidase to perform enzymatic hydrolysis. The resulting enzymatic hydrolysate is then ready for use. (5) Take the enzymatic hydrolysate described in step (4), filter it through a ceramic membrane, collect the first filtrate, filter it through an ultrafiltration membrane, collect the second filtrate, first enter the cation exchange resin column, and then enter the anion exchange resin column to obtain the second filtrate for later use. (6) Take the second effluent from step (5), concentrate it through nanofiltration membrane to obtain the first concentrate, then concentrate it under vacuum, add ethanol to the second concentrate, cool, filter and dry to obtain D-chiral inositol product.
2. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, In step (1), buckwheat flour and ethanol are added at a ratio of 1:10-16, and the volume concentration of ethanol is 30-70%.
3. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, In step (1), the temperature is heated to 30-60℃, the ultrasonic power is 300-500W, and the ultrasonic treatment time is 1-2h.
4. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, In step (2), the filtrate is concentrated until there is no alcohol odor, and purified water of 1-2 times the volume of the concentrate is added to the concentrate. The feed solution is fed into the resin column at a flow rate of 1-2 BV / h.
5. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, The solid content of the concentrate in step (3) is 25-35 wt%, and the pH is adjusted to 5.0-5.5 with glacial acetic acid; the brewing yeast and the concentrate are added at a dosage of 0.3-0.7 mg / ml, and the activity of the brewing yeast is 10^8-10^11 CFU / g; the fermentation temperature is 30-37℃, and the fermentation time is 2-3 days.
6. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, In step (4), 2-3 times the volume of purified water is added to the fermentation liquid. The enzyme activity of the α-galactosidase is 5000-8000 U / g, and the α-galactosidase is added at a dosage of 1-2 g / L.
7. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, The temperature of the enzymatic hydrolysis treatment in step (4) is 40-45℃, and the time of the enzymatic hydrolysis treatment is 5-8h.
8. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, The ceramic membrane in step (5) has a pore size of 150-300 nm and the ultrafiltration membrane has a molecular weight cutoff of 5000-10000 Da; the second filtrate enters the cation exchange resin column at a flow rate of 1-2 BV / h and the first effluent enters the anion exchange resin column at a flow rate of 0.5-1.0 BV / h.
9. The process for preparing D-chiral inositol from buckwheat according to claim 1, characterized in that, The nanofiltration membrane described in step (6) has a molecular weight cutoff of 150-300 Da; the solid content of the second concentrate is 40-50 wt%, and 0.7-0.8 times the volume of the second concentrate of ethanol is added, and the temperature is lowered to 20-25℃.