A green production process of diosgenin

Abstract

The present invention provides a green production process of diosgenin, which belongs to the field of biological extraction technology and includes the following steps: S1. crushing yellow ginger, adding water, mixing and filtering to obtain a filtrate; centrifuging the filtrate and collecting the centrifugal precipitate; adding water to the centrifugal precipitate for natural fermentation to obtain a fermentation liquid; S2. hydrolyzing the fermentation liquid with acid, adjusting the pH solution to neutrality and centrifuging, collecting the precipitate; extracting the precipitate by reflux with an organic solvent, filtering and concentrating and crystallizing; S3. further recrystallizing and purifying the crystal, then adding water and refluxing, filtering and drying to obtain diosgenin. The present invention provides a green production process of diosgenin, which can achieve the purpose of reducing costs and reducing the amount of organic solvents and acids and alkalis.

Description

Technical Field

[0001] The invention relates to the technical field of biological extraction, and in particular to a green production process of diosgenin. Background Art

[0002] Diosgenin is an organic intermediate and raw material for synthesizing many adrenocortical hormone drugs. It has many biological activities such as antibacterial, anti-inflammatory, and lipid-lowering, and is widely used in medicine. Turmeric is the plant with the most diosgenin in the world, and there is medical evidence that plant saponins are difficult to obtain through chemical synthesis. Therefore, in recent years, more and more companies have used turmeric as a raw material to produce diosgenin. At present, the industrial methods of producing diosgenin using turmeric as a raw material have their own advantages and disadvantages. The traditional diosgenin production method, such as the organic solvent extraction method, is to extract diosgenin with organic solvents such as petroleum ether and gasoline, and then use a large amount of acid or biological enzymes to break the glycosidic bond to free diosgenin, and finally achieve the effect of extracting diosgenin. This process is currently the most used process in production, but each ton of diosgenin production consumes a large amount of acid and alkali, and is difficult to recycle, and a large amount of industrial wastewater is discharged, which puts considerable pressure on the environment; if it is produced with biological enzymes, it will greatly increase the production cost.

[0003] The patent document with publication number CN107501384A and titled "Method for Clean Production of Diosgenin Using Turmeric" provides a method for large-scale production of turmeric saponin from turmeric rhizomes using extraction, acid hydrolysis, crystallization and recrystallization. At the same time, the waste is used to produce a large amount of modified starch, and cellulose is effectively used to produce water-retaining materials. However, organic substances such as N-methylmorpholine-N-oxide, acrylic acid, and glutaraldehyde are used in the process, which makes the final product carry the risk of excessive solvent residues, and there are certain challenges in product quality control.

[0004] The patent document with publication number CN105925559A and titled "Preparation of an immobilized enzyme and a method for hydrolyzing diosgenin" discloses that pullulanase, fungal α-amylase, rhamnosidase, β-glucosidase and other biological enzymes are immobilized on a carrier using dextran or macroporous adsorption resin to prepare an immobilized enzyme preparation composition. Diosgenin is then enzymolyzed with the immobilized enzyme preparation composition to produce diosgenin. This method uses expensive biological enzymes, and the immobilization of enzymes requires special equipment, which increases the production cost of diosgenin and restricts its large-scale production.

[0005] Therefore, it is necessary to provide a green production process of diosgenin to solve the problems existing in the above-mentioned prior art. Summary of the invention

[0006] In view of this, the present invention provides a green production process of diosgenin, which can reduce the cost and the amount of organic solvents, acids and alkalis used.

[0007] To achieve the above object, the present invention provides a green production process of diosgenin, comprising the following steps: S1. Grind the turmeric, add water, mix well and filter to obtain a filtrate; centrifuge the filtrate and collect the centrifugal precipitate; add water to the centrifugal precipitate and ferment it naturally to obtain a fermentation liquid; S2. The fermentation broth is hydrolyzed by acid, the pH of the solution is adjusted to neutral, and then centrifuged to collect the precipitate; the precipitate is extracted by reflux with an organic solvent, filtered, concentrated and crystallized; S3. The crystals are further recrystallized and purified, then water is added for reflux, filtered and dried to obtain diosgenin.

[0008] Optionally, the centrifugal conditions are a rotation speed of 4800-5000 r / min and a time of 12-20 min.

[0009] Optionally, S1 includes cutting fresh turmeric into thin slices and then drying, grinding into fine powder, adding water and mixing evenly, sieving, collecting liquid, adding water to the solid residue and stirring evenly, sieving again, combining the two liquids to obtain a filtrate; centrifuging the filtrate and collecting the centrifugal precipitate; adding water to the centrifugal precipitate for natural fermentation to obtain a fermentation liquid.

[0010] Optionally, the ratio of turmeric to water when adding water and mixing evenly is 1:4-5, the ratio of solid residue to water when adding water and mixing evenly again is 1:0.2-0.6, and the ratio of centrifugal precipitation to water during natural fermentation is 1:1-2.

[0011] Optionally, the sieving is a 80-120 mesh sieve, and the natural fermentation time is 45-50 hours.

[0012] Optionally, hydrochloric acid is used for acid hydrolysis in S2, and lime water is used for adjusting pH.

[0013] Optionally, the hydrochloric acid concentration is 0.2 mol / L, and the hydrolysis condition is to maintain the reflux state for 4.5 to 5.5 hours.

[0014] Optionally, the organic solvent reflux extraction conditions in S2 are: solid-liquid ratio of 1:10, extraction time of 4 hours, extraction temperature of 85° C., and extraction twice; the organic solvent is petroleum ether or 120# gasoline.

[0015] Optionally, when water is added to S3 for reflux, the temperature is 100° C., the material-liquid ratio is 1:9-11, and the time is 1.5-2.5 h.

[0016] In order to achieve the above object, the present invention also provides diosgenin prepared by the green production process of the above diosgenin.

[0017] The above technical solution of the present invention includes at least the following beneficial effects: The technical solution provided by the present invention can save 40% of the acid usage, thereby reducing the usage of alkali. According to liquid chromatography detection, the diosgenin prepared by the present invention has high purity and little residual organic solvent.

[0018] The technical solution provided by the present invention reduces the amount of acid and alkali used, and the raw materials used have low process costs compared to biological enzymes in the prior art. At the same time, the discharge of wastewater is reduced, the process is environmentally friendly and simple, and it is easy to expand scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is a high performance liquid chromatogram of diosgenin, a product in Example 1 of the present invention; Figure 2 The high performance liquid chromatogram of the product diosgenin in Comparative Example 1 of the present invention. DETAILED DESCRIPTION

[0020] To make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the following will be combined with the appended drawings of the embodiments of the present invention. Figure 1~2 , the technical scheme of the embodiment of the present invention is clearly and completely described. Obviously, the described embodiment is a part of the embodiment of the present invention, not all of the embodiments. Based on the described embodiment of the present invention, all other embodiments obtained by ordinary technicians in this field belong to the scope of protection of the present invention.

[0021] Embodiment 1 (1) Cut fresh turmeric into thin slices, dry at 80°C, grind into fine powder, weigh 100g of turmeric powder, add 400mL of water, stir evenly, pass through an 80-mesh sieve, collect the liquid, add 50mL of water to the solid residue, stir evenly, pass through an 80-mesh sieve, and combine the two liquids for later use; (2) Centrifuge the above liquid at 4800 r / min for 12 min in a high-speed centrifuge, collect the centrifuged precipitates and combine them for later use; (3) Add water to the centrifugal precipitate, with the ratio of precipitate to water being 1:2, and place it in a constant temperature box at 40°C for 2 days for fermentation.

[0022] (4) Add hydrochloric acid to the fermentation broth to adjust the concentration to 0.2 mol / L, heat and hydrolyze under reflux for 5 h, cool to room temperature, add lime water to adjust the pH of the mixture to neutral, collect the solid precipitate by centrifugation, dry in an oven at 80°C, and grind into fine powder; (5) Add petroleum ether to the above powder at a solid-liquid ratio of 1:10, reflux extract at 85°C for 2 times, each time for 4 hours, filter and collect the liquid, concentrate and crystallize, filter the crystals, and dry at 80°C; (6) further recrystallization, filtering out the crystals, drying and crushing; (7) The crystals were added to water at a solid-liquid ratio of 1:10, refluxed at 100°C for 2 h, filtered while hot, and dried to obtain the final product, diosgenin. The product was tested by high performance liquid chromatography (HPLC), and the purity of the product was 98%.

[0023] Embodiment 2 (1) Cut fresh turmeric into thin slices and dry at 80°C. Weigh 500g of turmeric powder, add 2500mL of water, stir evenly, pass through a 120-mesh sieve, collect the liquid, add 100mL of water to the solid residue, stir evenly, pass through a 120-mesh sieve, and combine the two liquids for later use; (2) Centrifuge the above liquid at 5000 r / min for 15 min in a high-speed centrifuge, collect the centrifuged precipitates and combine them for later use; (3) Add water to the centrifugal precipitate, with the precipitate to water ratio being 1:1, and place in a 40°C constant temperature box for fermentation for 45 hours.

[0024] (4) Add hydrochloric acid to the fermentation broth to adjust the concentration to 0.2 mol / L, hydrolyze under reflux for 4.5 h, cool to room temperature, add lime water to adjust the pH of the mixture to neutral, collect the solid precipitate by centrifugation, dry in an oven at 80°C, and grind into fine powder; (5) Add petroleum ether to the above powder at a solid-liquid ratio of 1:10, reflux extract at 85°C for 2 times, each time for 4 hours, collect the liquid by filtration, concentrate and crystallize, filter the crystals by suction, and dry at 80°C; (6) further recrystallization, filtering out the crystals, drying and crushing; (7) The crystals were added to water at a solid-liquid ratio of 1:10, refluxed at 100°C for 2.5 h, filtered while hot, and dried to obtain the final product, diosgenin. The product purity was 98% as determined by HPLC.

[0025] Embodiment 3 (1) Cut fresh turmeric into thin slices, dry at 80°C, grind into fine powder, weigh 1000g of turmeric powder, add 4500mL of water, stir evenly, pass through a 100-mesh sieve, collect the liquid, add 600mL of water to the solid residue, stir evenly, pass through a 100-mesh sieve, and combine the two liquids for later use; (2) Centrifuge the above liquid at 4900 r / min for 18 min in a high-speed centrifuge, collect the centrifuged precipitates and combine them for later use; (3) Add the above centrifugal precipitate to water with a solid-liquid ratio of precipitate to water of 1:2, and ferment in a constant temperature box at 40°C for 50 hours.

[0026] (4) Add hydrochloric acid to the fermentation broth to adjust the concentration to 0.2 mol / L, heat and hydrolyze under reflux for 5.5 h, cool to room temperature, add lime water to adjust the pH of the mixture to neutral, collect the solid precipitate by centrifugation, dry in an oven at 80°C, and grind into fine powder; (5) Add 120# gasoline to the above powder with a solid-liquid ratio of 1:10, reflux extract twice at 85°C, each time for 4 hours, filter and collect the liquid, concentrate and crystallize, filter the crystals, and dry at 80°C; (6) further recrystallization, filtering out the crystals, drying and crushing; (7) The crystals were added to water at a solid-liquid ratio of 1:11, refluxed at 100°C for 2 h, filtered while hot, and dried to obtain the final product, diosgenin. The purity of the product was 99% as determined by HPLC.

[0027] Embodiment 4 (1) Cut fresh turmeric into thin slices and dry at 80°C, grind into fine powder, weigh 10 kg of turmeric powder, add 40 L of water, stir evenly, pass through an 80-mesh sieve, collect the liquid, add 5 L of water to the solid residue, stir evenly, pass through an 80-mesh sieve, and combine the two liquids for later use; (2) Centrifuge the above liquid at 4800 r / min for 20 min in a high-speed centrifuge, collect the centrifuged precipitates and combine them for later use; (3) Add water to the centrifugal precipitate, with the precipitate to water ratio being 1:1, and place it in a constant temperature box at 40°C for 2 days for fermentation.

[0028] (4) Add hydrochloric acid to the fermentation broth to adjust the concentration to 0.2 mol / L, heat and hydrolyze under reflux for 5 h, cool to room temperature, add lime water to adjust the pH of the mixture to neutral, collect the solid precipitate by centrifugation, dry in an oven at 80°C, and grind into fine powder; (5) Add 120# gasoline to the above powder with a solid-liquid ratio of 1:10, reflux extract twice at 85°C, each time for 4 hours, filter and collect the liquid, concentrate and crystallize, filter the crystals, and dry at 80°C; (6) further recrystallization, filtering out the crystals, drying and crushing; (7) Add 10 times the amount of water to the crystals and reflux at 100°C for 2 h. Filter while hot and dry to obtain the final product, diosgenin. The purity of the product was 98% as determined by HPLC.

[0029] Comparative Example 1 (1) Cut fresh turmeric into thin slices, dry at 80°C, grind into fine powder, weigh 100g of turmeric powder, add 400mL of water, stir evenly, pass through an 80-mesh sieve, collect the liquid, add 50mL of water to the solid residue, stir evenly, pass through an 80-mesh sieve, and combine the two liquids for later use; (2) Centrifuge the above liquid at 4800 r / min for 12 min in a high-speed centrifuge, collect the centrifuged precipitates and combine them for later use; (3) Add twice the amount of water to the above centrifugal sediment and ferment it in a constant temperature box at 40℃ for 2 days.

[0030] (4) Add hydrochloric acid to the fermentation broth to adjust the concentration to 0.2 mol / L, heat and hydrolyze under reflux for 5 h, cool to room temperature, add lime water to adjust the pH of the mixture to neutral, collect the solid precipitate by centrifugation, dry in an oven at 80°C, and grind into fine powder; (5) Add petroleum ether to the above powder and reflux extract at 85°C for 2 times, each time for 4 hours, collect the liquid by filtration, concentrate and crystallize, filter the crystals by suction, and dry at 80°C; (6) Further recrystallization, filtering out crystals, drying and crushing to obtain the final product diosgenin. HPLC detection showed that the purity of the product was 95%.

[0031] Comparative Example 2 (1) Cut fresh turmeric into thin slices, dry at 80°C, grind into fine powder, weigh 100g of turmeric powder, add 400mL of water, stir evenly, pass through an 80-mesh sieve, collect the liquid, add 50mL of water to the solid residue, stir evenly, pass through an 80-mesh sieve, and combine the two liquids for later use; (2) Centrifuge the above liquid at 4800 r / min for 12 min in a high-speed centrifuge, collect the centrifuged precipitates and combine them for later use; (3) adding hydrochloric acid to the above centrifuged precipitate to a hydrochloric acid concentration of 0.2 mol / L, heating and hydrolyzing under reflux for 5 h, cooling to room temperature, adding lime water to adjust the pH of the mixture to neutral, centrifuging and collecting the solid precipitate, drying in an oven at 80°C, and grinding into fine powder; (4) Add petroleum ether to the above powder at a solid-liquid ratio of 1:10, reflux extract twice at 85°C, each time for 4 hours, filter and collect the liquid, concentrate and crystallize, filter the crystals, and dry at 80°C; (5) further recrystallization, filtering out the crystals, drying and crushing; (6) The crystals were added to water at a solid-liquid ratio of 1:10, refluxed at 100°C for 2 h, filtered while hot, and dried to obtain the final product, diosgenin. The purity of the product was 90% as determined by HPLC.

[0032] The diosgenin prepared in Examples 1 to 4 and Comparative Examples 1 to 2 was tested by HPLC, and the purity of the product is shown in Table 1. The HPLC chromatograms of Example 1 and Comparative Example 1 are shown in Figure 1 , Figure 2 .

[0033] Table 1 Purity of diosgenin prepared in Examples 1 to 4 and Comparative Examples 1 to 2

[0034] It can be seen from Table 1 that the purity of diosgenin prepared in the embodiment of the present invention is higher than that of Comparative Examples 1 and 2.

[0035] Depend on Figure 1 , 2 Combining the analysis of Example 1 and Comparative Example 1, the difference between Comparative Example 1 and Example 1 is that the step of adding water and reflux is not performed. Figure 1 It shows that the diosgenin prepared in Example 1 has high purity and few impurities. Figure 2 It is shown that the diosgenin prepared in Comparative Example 1 contains many impurities. It can be seen that the step of adding water and refluxing the crystals after recrystallization in the present invention can reduce the impurities in the product and improve the purity of the product.

[0036] From the analysis of Table 1 in combination with Example 1 and Comparative Example 2, the only difference between Comparative Example 2 and Example 1 is that the natural fermentation step is not performed, and the purity of diosgenin prepared in Comparative Example 2 is lower than the purity of diosgenin prepared in Example 1. It can be seen that the natural fermentation step in the present invention improves the purity of the product.

[0037] The above are preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A green production process of diosgenin, characterized in that: The following steps are involved: S1. Grind the turmeric, add water, mix well and filter to obtain a filtrate; centrifuge the filtrate and collect the centrifugal precipitate; add water to the centrifugal precipitate and ferment it naturally to obtain a fermentation liquid; S2. The fermentation broth is hydrolyzed by acid, the pH of the solution is adjusted to neutral, and then centrifuged to collect the precipitate; the precipitate is extracted by reflux with an organic solvent, filtered, concentrated and crystallized; S3. The crystals are further recrystallized and purified, then water is added for reflux, filtered and dried to obtain diosgenin.

2. The green production process of diosgenin according to claim 1, characterized in that: The centrifugal conditions are a rotation speed of 4800-5000 r / min and a time of 12-20 min.

3. The green production process of diosgenin according to claim 1, characterized in that: The step S1 comprises cutting fresh turmeric into thin slices, drying the slices, grinding the slices into fine powder, adding water to mix well, sieving, collecting liquid, adding water to the solid residue, mixing well, sieving again, combining the two liquids to obtain a filtrate; centrifuging the filtrate, collecting a centrifugal precipitate; adding water to the centrifugal precipitate and fermenting it naturally to obtain a fermentation liquid.

4. The green production process of diosgenin according to claim 3, characterized in that: When adding water and mixing evenly, the ratio of turmeric to water is 1:4-5, when adding water and mixing evenly again, the ratio of solid residue to water is 1:0.2-0.6, and during the natural fermentation, the ratio of centrifugal precipitation to water is 1:1-2.

5. The green production process of diosgenin according to claim 3, characterized in that: The sieving is a 80-120 mesh sieve, and the natural fermentation time is 45-50 hours.

6. The green production process of diosgenin according to claim 1, characterized in that: Hydrochloric acid is used for acid hydrolysis in S2, and lime water is used for pH adjustment.

7. The green production process of diosgenin according to claim 6, characterized in that: The hydrochloric acid concentration is 0.2 mol / L, and the hydrolysis condition is to maintain the reflux state for 4.5 to 5.5 hours.

8. The green production process of diosgenin according to claim 1, characterized in that: The conditions for the reflux extraction of the organic solvent in S2 are: a solid-liquid ratio of 1:10, an extraction time of 4 hours, an extraction temperature of 85° C., and extractions twice; the organic solvent is petroleum ether or 120# gasoline.

9. The green production process of diosgenin according to claim 1, characterized in that: When water is added to S3 for reflux, the temperature is 100° C., the material-liquid ratio is 1:9-11, and the time is 1.5-2.5 h.

10. Diosgenin obtained by the green production process of diosgenin according to any one of claims 1 to 9.

Images