Method for extracting and purifying dinorhydrin

Abstract

The invention relates to an extraction and purification method for double-norhydrin, and belongs to the technical field of medicine production.The extraction and purification method for double-norhydrin comprises the steps that a filter cake obtained after fermentation and filtration makes direct contact with methyl alcohol, the methyl alcohol is used for extraction, the methyl alcohol and water are mutually soluble, a solvent makes contact with a product more easily, and the extraction efficiency of the product is improved; protein extracted in the extraction process can be hydrolyzed, the influence of the protein on filtration can be solved, when alkali is not adopted, the filtration process cannot be normally carried out, and filtration can be difficultly completed only by continuously replacing a filtration medium, and when alkali is adopted, a crude product can be easily obtained through one-time filtration, and a sodium hydroxide solution is adopted for impurity removal; the moisture content of a final wet product is 30-40%. And the final product drying efficiency is effectively improved.

Description

Technical Field

[0001] The invention belongs to the technical field of drug production, and in particular relates to a method for extracting and purifying dihydrochloride. Background Art

[0002] Steroid drugs are a large class of drugs with many varieties and wide applications. Their clinical demand is second only to antibiotics. They are mainly used to treat rheumatism, cardiovascular, collagen diseases, lymphatic leukemia, tumors, bacterial encephalitis, skin diseases, endocrine disorders, senile diseases, etc. The English name of bispregnol (21-hydroxy-20-methylpregn-4-en-3-one); CAS number 60966-36-1; molecular formula: C 22 H 34 O2; molecular weight: 330.50; structural formula: Dihydrotestosterone is the starting material of steroid hormones and is used as the starting material of raw materials such as progesterone and ursocholic acid. Dihydrotestosterone is an important intermediate of steroid hormone drugs and has important commercial value. In the prior art, the preparation of dihydrotestosterone generally requires drying the filter cake and then extracting it with solvents such as methanol and ethyl acetate. The operation is complicated, and the yield of dihydrotestosterone is not high. The solvent used to extract dihydrotestosterone cannot be effectively recovered, resulting in a waste of resources. Summary of the invention

[0003] In order to overcome the problems in the background technology, the present invention discloses a method for extracting and purifying double-decanoic acid alcohol. In the method for extracting and purifying double-decanoic acid alcohol, the filter cake after fermentation filtration is directly contacted with methanol, and methanol is used for extraction. Methanol and water are miscible, and the solvent is more likely to contact the product, thereby improving the extraction efficiency of the product. By adding alkali to the crude product concentrate, the protein extracted in the extraction process can be hydrolyzed, and the influence of the protein on the filtration can also be solved. When the alkali is not used, the filtration process cannot be carried out normally, and the filtration medium needs to be continuously replaced to complete the filtration with difficulty. When the alkali is used, the crude product can be easily obtained by one-time filtration.

[0004] To achieve the above object, the present invention is implemented through the following technical solutions: The method for extracting and purifying the double-decanoic acid alcohol is characterized in that it comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution; (2) Inoculate the seed solution with Mycobacterium neoaureum at a rate of 1%-3% for cultivation, and keep the qualified ones for future use; (2) Prepare transformation medium; (3) Inoculate the qualified seed liquid into the transformation medium at an inoculum rate of 5%-15%, add 30 g / L of phytosterols to the medium, and carry out transformation and fermentation for 3-4 days. When the phytosterol conversion rate is ≥95%, stop the fermentation and obtain the dihydrogen ester fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The filter cake is obtained by hydraulic filtration of the double alcohol fermentation; (2) Extraction: After the filter cake is added to the extraction tank, it is extracted with methanol and filtered to obtain the extract; (3) Decolorization of the extract: After the extract is added to the decolorization tank, activated carbon is added for decolorization and filtration to obtain a decolorized liquid; (4) Distillation of decolorizing liquid: The decolorizing liquid is distilled at normal pressure, and the distillate is collected in sections, and the residual distillate is left; (5) treating the distilled residual liquid with alkali to obtain a crude dihydrochloride; 3) Purification after extraction of the crude product of dihydrochloride: the crude product purification comprises the following steps: (1) Purification of crude dihydrochloride: The crude dihydrochloride is put into a purification tank, and methanol aqueous solution is added for purification to obtain a purified dihydrochloride crude product liquid; (2) Treatment of the purified liquid of crude dihydrochloride: adding alkali to the purified liquid of crude dihydrochloride to adjust the solution, obtain crystals and filter them; (3) Crystallization treatment: washing with drinking water to obtain a purified product of dihydrochloric acid; (4) re-purification: re-purifying the first purified product of dihydrochloride according to the methods of steps (1), (2) and (3) to obtain a second purified product of dihydrochloride; 4) Post-processing: The secondary purified product of dihydrochloride is dried and then crushed and packaged.

[0005] Preferably, the seed solution in step 1) is qualified when the seed OD value reaches 10-12.

[0006] Preferably, in step 2), the extraction operation is to add methanol 3 to 8 times the mass of the precursor phytosterol, stir at room temperature for 1 to 3 hours for extraction, and then filter using the filter medium in the extraction tank to obtain the extract.

[0007] Preferably, the extraction operation in step 2) is repeated at least four times until the filter cake is completely extracted, and the degree of extraction of the filter cake is detected by TLC. Preferably, the decolorization operation in step 2) is to add activated carbon with a mass of 0.05-0.2 times of the mass of phytosterol to the extract, raise the temperature to 50-60°C, stir and decolorize for 1-3 hours, and distill after the decolorization is completed to an internal temperature of 72-74°C.

[0008] Preferably, in step 2), the distilled solvent is recovered and reused, and the solvent with a concentration lower than 40% is separately collected and processed to obtain methanol that can be recovered and reused.

[0009] Preferably, in step 2), the distillation residue is added with alkali to adjust the pH value of the distillation residue to 8.5-9.0 using 40% sodium hydroxide solution, the temperature is raised and refluxed for 1-3 hours, the temperature is lowered to 15-20° C. and stirred for crystallization for 2-10 hours, and the crude dihydrochloride is obtained by filtration.

[0010] Preferably, in step 3), a 40%-70% methanol aqueous solution of 1.5-3 times the mass of phytosterol is added to purify the crude product of bis-decanoic acid, and the mixture is stirred and heated to 60-70°C to purify the crude product to obtain a bis-decanoic acid crude product purified solution; the bis-decanoic acid crude product purified solution is treated by adjusting the pH value to 8.5-9.0 with a 40% sodium hydroxide solution, heating and refluxing for 1-3 hours, cooling to 15-20°C, stirring and crystallizing for 2-10 hours, and filtering to obtain a crystal; Preferably, in step 3), the crystals are washed with drinking water having a pH value of 6.0-6.5, and washing is stopped when the outlet pH value reaches the pH value of drinking water, and the filtration is continued until there are no droplets, and the crystals are dried for 1-3 hours to obtain a purified product of dihydrochloride; Preferably, in step 4), the post-treatment is to put the secondary purified product of dihydrogen sulfoxide into a single cone dryer at a temperature of 90-100° C. for 12-24 hours, then cool it to room temperature, crush it with a universal crusher, and then pack it in a fixed amount.

[0011] Beneficial effects of the present invention: The double alcohol extraction and purification method directly contacts the filter cake after fermentation filtration with methanol, and uses methanol for extraction. Methanol and water are mutually soluble, and the solvent is more likely to contact the product, thereby improving the extraction efficiency of the product. By adding alkali to the crude product concentrate, the protein extracted in the extraction process can be hydrolyzed, and the influence of protein on filtration can also be solved. When alkali is not used, the filtration process cannot be carried out normally, and the filtration medium needs to be continuously replaced to complete the filtration with great difficulty. When alkali is used, the crude product can be easily obtained by one-time filtration, and sodium hydroxide solution is used for impurity removal. The moisture content of the final product wet product is 30-40%. The drying efficiency of the final product is effectively improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 Detection spectrum of dihydrochloride DETAILED DESCRIPTION

[0013] In order to make the above-mentioned objectives, technical solutions and beneficial effects clearer, the present invention is described in detail below in conjunction with embodiments.

[0014] Example 1 A method for extracting and purifying bisphenol A comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution: mix 20 g / L glycerol, 8 g / L (NH4)2HPO4, 0.5 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1 g / L corn steep liquor powder (Angel), and 3 g / L defoamer (GPE), add distilled water to make up to 1000 mL, and then sterilize. After sterilization, the pH value is 7.0. Inoculate 2% of the volume of the seed solution with Mycobacterium neoaureum for cultivation for 30 h. When the OD value of the seed solution reaches 12, it is ready for use. (2) Prepare transformation medium: 20 g / L glucose, 8 g / L ammonium citrate tribasic, 0.7 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1.5 g / L Tween 80, 3 g / L lecithin (PC40), 3 g / L defoaming agent (GPE), 30 g / L phytosterol (800 mesh). After the above materials are prepared, sterilize them for use. The pH after sterilization is 7.0. (3) Inoculate the qualified seed liquid into the transformation medium at a rate of 15%, and carry out transformation and fermentation for 4 days. When the phytosterol conversion rate reaches ≥95%, stop the fermentation and obtain the phytosterol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The fermentation liquid of the double alcohol fermentation is filtered through a box filter press to obtain a filter cake containing bacteria and products. The filtrate enters the wastewater treatment network as general fermentation wastewater; (2) Extraction: After the filter cake obtained by filtration is added to an extraction tank, methanol 3 times the mass of the precursor substance phytosterol is added to the extraction tank for extraction, and the extract is obtained by filtration; the extraction is carried out by stirring at room temperature for 1 hour, and then the extraction is carried out by filtering using the filter medium in the extraction tank to obtain an extract. The extraction operation is repeated at least four times until the filter cake is completely extracted, and the extracts of multiple times are combined; (3) Decolorization of the extract: add activated carbon in an amount 0.05 times the mass of the phytosterols to the extract, raise the temperature to 50°C, stir and decolorize for 1 hour to obtain a decolorized solution, and distill the decolorized solution at normal pressure until the internal temperature reaches 72°C. Collect the distillate in sections, and leave the residual distillate. (4) cooling the residual liquid to 15-20°C, stirring and crystallizing for 2-10 hours, and filtering to obtain a crude product of dihydrochloride; 3) Extracting and purifying the crude product of dihydrochloride: (1) Purification of crude bisphenol A: The crude bisphenol A was put into a purification tank, and methanol-water solution was added for purification to obtain a crude bisphenol A purified solution; a 40% methanol-water solution with a mass of 1.5 times that of phytosterol was added, and the mixture was stirred and heated to 60°C for purification to obtain a crude bisphenol A purified solution; the purified bisphenol A was filtered until there were no droplets, and dried for 1 hour to obtain a bisphenol A extract. The bisphenol A extract was put into a single cone dryer at 90°C for 12 hours, and then cooled to room temperature. It was crushed with a universal grinder and packaged in a fixed amount.

[0015] Example 2 A method for extracting and purifying bisphenol A comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution: mix 20 g / L glycerol, 8 g / L (NH4)2HPO4, 0.5 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1 g / L corn steep liquor powder (Angel), and 3 g / L defoamer (GPE), add distilled water to make up to 1000 mL, and then sterilize. After sterilization, the pH value is 7.0. Inoculate 2% of the volume of the seed solution with Mycobacterium neoaureum for cultivation for 30 h. When the OD value of the seed solution reaches 12, it is ready for use. (2) Prepare transformation medium: 20 g / L glucose, 8 g / L ammonium citrate tribasic, 0.7 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1.5 g / L Tween 80, 3 g / L lecithin (PC40), 3 g / L defoaming agent (GPE), 30 g / L phytosterol (800 mesh). After the above materials are prepared, sterilize them for use. The pH after sterilization is 7.0. (3) Inoculate the qualified seed liquid into the transformation medium at a rate of 15%, and carry out transformation and fermentation for 4 days. When the phytosterol conversion rate reaches ≥95%, stop the fermentation and obtain the phytosterol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The fermentation liquid of the double alcohol fermentation is filtered through a box filter press to obtain a filter cake containing bacteria and products. The filtrate enters the wastewater treatment network as general fermentation wastewater; (2) Extraction: After the filter cake obtained by filtration is added to an extraction tank, methanol 3 to 8 times the mass of the precursor substance phytosterol is added to the extraction tank for extraction, and the extract is obtained by filtration; the extraction is carried out by stirring at room temperature for 2 hours, and then the extraction is carried out by filtering using the filter medium in the extraction tank to obtain an extract. The extraction operation is repeated at least four times until the filter cake is completely extracted, and the multiple extracts are combined; (3) Decolorization of the extract: add activated carbon with a mass of 0.1 times the mass of the phytosterol to the extract, raise the temperature to 55°C, stir and decolorize for 2 hours to obtain a decolorized liquid, and distill the decolorized liquid at normal pressure until the internal temperature reaches 73°C. Collect the distillate in sections, and leave the residual distillate; (4) Cooling the distilled residual liquid to 17°C, stirring and crystallizing for 6 hours, and filtering to obtain a crude product of dihydrochloride; 3) Extracting and purifying the crude product of dihydrochloride: (1) Purification of crude bisphenol A: The crude bisphenol A was put into a purification tank, and methanol-water solution was added for purification to obtain a crude bisphenol A purified solution; 55% methanol-water solution with a mass twice that of phytosterol was added, and the mixture was stirred and heated to 65°C for purification to obtain a crude bisphenol A purified solution; the purified bisphenol A was filtered until there were no droplets, and dried for 2 hours to obtain a bisphenol A extract; the bisphenol A extract was put into a single cone dryer at 95°C for 20 hours, and then cooled to room temperature, crushed with a universal grinder, and packaged in a fixed amount.

[0016] Example 3 A method for extracting and purifying bisphenol A comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution: mix 20 g / L glycerol, 8 g / L (NH4)2HPO4, 0.5 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1 g / L corn steep liquor powder (Angel), and 3 g / L defoamer (GPE), add distilled water to make up to 1000 mL, and then sterilize. After sterilization, the pH value is 7.0. Inoculate 2% of the volume of the seed solution with Mycobacterium neoaureum for cultivation for 30 h. When the OD value of the seed solution reaches 12, it is ready for use. (2) Prepare transformation medium: 20 g / L glucose, 8 g / L ammonium citrate tribasic, 0.7 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1.5 g / L Tween 80, 3 g / L lecithin (PC40), 3 g / L defoaming agent (GPE), 30 g / L phytosterol (800 mesh). After the above materials are prepared, sterilize them for use. The pH after sterilization is 7.0. (3) Inoculate the qualified seed liquid into the transformation medium at a rate of 15%, and carry out transformation and fermentation for 4 days. When the phytosterol conversion rate reaches ≥95%, stop the fermentation and obtain the phytosterol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The fermentation liquid of the double alcohol fermentation is filtered through a box filter press to obtain a filter cake containing bacteria and products. The filtrate enters the wastewater treatment network as general fermentation wastewater; (2) Extraction: After the filter cake obtained by filtration is added to an extraction tank, methanol 3 to 8 times the mass of the precursor substance phytosterol is added to the extraction tank for extraction, and the extract is obtained by filtration; the extraction is carried out by stirring at room temperature for 3 hours, and then the extraction is carried out by filtering using the filter medium in the extraction tank to obtain an extract. The extraction operation is repeated at least four times until the filter cake is completely extracted, and the multiple extracts are combined; (3) Decolorization of the extract: add activated carbon 0.2 times the mass of the phytosterol to the extract, raise the temperature to 60°C, stir and decolorize for 3 hours to obtain a decolorized solution, and distill the decolorized solution at normal pressure until the internal temperature reaches 74°C. Collect the distillate in sections, and leave the residual distillate. (4) Cooling the residual liquid to 20°C, stirring and crystallizing for 10 hours, and filtering to obtain a crude product of dihydrochloride; 3) Extracting and purifying the crude product of dihydrochloride: (1) Purification of crude bisphenol A: The crude bisphenol A was put into a purification tank, and methanol-water solution was added for purification to obtain a crude bisphenol A purified solution; 70% methanol-water solution with a mass of 3 times that of phytosterol was added, and the mixture was stirred and heated to 70°C for purification to obtain a crude bisphenol A purified solution; the purified bisphenol A was filtered until there were no droplets, and dried for 3 hours to obtain a bisphenol A extract. The bisphenol A extract was put into a single cone dryer at 100°C for 24 hours, and then cooled to room temperature. It was crushed with a universal grinder and packaged in a fixed amount.

[0017] Example 4 A method for extracting and purifying bisphenol A comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution: mix 20 g / L glycerol, 8 g / L (NH4)2HPO4, 0.5 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1 g / L corn steep liquor powder (Angel), and 3 g / L defoamer (GPE), add distilled water to make up to 1000 mL, and then sterilize. After sterilization, the pH value is 7.0. Inoculate 2% of the volume of the seed solution with Mycobacterium neoaureum for cultivation for 30 h. When the OD value of the seed solution reaches 12, it is ready for use. (2) Prepare transformation medium: 20 g / L glucose, 8 g / L ammonium citrate tribasic, 0.7 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1.5 g / L Tween 80, 3 g / L lecithin (PC40), 3 g / L defoaming agent (GPE), 30 g / L phytosterol (800 mesh). After the above materials are prepared, sterilize them for use. The pH after sterilization is 7.0. (3) Inoculate the qualified seed liquid into the transformation medium at a rate of 15%, and carry out transformation and fermentation for 4 days. When the phytosterol conversion rate reaches ≥95%, stop the fermentation and obtain the phytosterol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The fermentation liquid of the double alcohol fermentation is filtered through a box filter press to obtain a filter cake containing bacteria and products. The filtrate enters the wastewater treatment network as general fermentation wastewater; (2) Extraction: After the filter cake obtained by filtration is added to an extraction tank, methanol in an amount 3 to 8 times the mass of the precursor substance phytosterol is added to the extraction tank for extraction, and the extract is obtained by filtration; the extraction is carried out by stirring at room temperature for 2.5 hours, and then the extraction is carried out by filtering using the filter medium in the extraction tank to obtain an extract. The extraction operation is repeated at least four times until the filter cake is completely extracted, and the extracts of multiple times are combined; (3) Decolorization of the extract: add activated carbon in an amount 0.15 times the mass of the phytosterols to the extract, raise the temperature to 58°C, stir and decolorize for 2.5 hours to obtain a decolorized solution, and distill the decolorized solution at normal pressure until the internal temperature reaches 70°C. Collect the distillate in sections, and leave the residual distillate. (4) cooling the residual liquid to 18°C, stirring and crystallizing for 8 hours, and filtering to obtain a crude product of dihydrochloride; 3) Extracting and purifying the crude product of dihydrochloride: (1) Purification of crude bisphenol A: The crude bisphenol A was put into a purification tank, and methanol-water solution was added for purification to obtain a crude bisphenol A purified solution; 65% methanol-water solution with a mass of 2.5 times that of phytosterol was added, and the mixture was stirred and heated to 67°C for purification to obtain a crude bisphenol A purified solution; the purified bisphenol A was filtered until there were no droplets, and dried for 2.5 hours to obtain a bisphenol A extract. The bisphenol A extract was put into a single cone dryer at 98°C for 22 hours, and then cooled to room temperature. It was crushed with a universal grinder and packaged in a fixed amount.

[0018] Example 5 A method for extracting and purifying bisphenol A comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution: mix 20 g / L glycerol, 8 g / L (NH4)2HPO4, 0.5 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1 g / L corn steep liquor powder (Angel), and 3 g / L defoamer (GPE), add distilled water to make up to 1000 mL, and then sterilize. After sterilization, the pH value is 7.0. Inoculate 2% of the volume of the seed solution with Mycobacterium neoaureum for cultivation for 30 h. When the OD value of the seed solution reaches 12, it is ready for use. (2) Prepare transformation medium: 20 g / L glucose, 8 g / L ammonium citrate tribasic, 0.7 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1.5 g / L Tween 80, 3 g / L lecithin (PC40), 3 g / L defoaming agent (GPE), 30 g / L phytosterol (800 mesh). After the above materials are prepared, sterilize them for use. The pH after sterilization is 7.0. (3) Inoculate the qualified seed liquid into the transformation medium at a rate of 15%, and carry out transformation and fermentation for 4 days. When the phytosterol conversion rate reaches ≥95%, stop the fermentation and obtain the phytosterol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The fermentation liquid of the double alcohol fermentation is filtered through a box filter press to obtain a filter cake containing bacteria and products. The filtrate enters the wastewater treatment network as general fermentation wastewater; (2) Extraction: After the filter cake obtained by filtration is added to an extraction tank, methanol 3 to 8 times the mass of the precursor substance phytosterol is added to the extraction tank for extraction, and the extract is obtained by filtration; the extraction is carried out by stirring at room temperature for 3 hours, and then the extraction is carried out by filtering using the filter medium in the extraction tank to obtain an extract. The extraction operation is repeated at least four times until the filter cake is completely extracted, and the multiple extracts are combined; (3) Decolorization of the extract: add activated carbon 0.2 times the mass of the phytosterol to the extract, raise the temperature to 60°C, stir and decolorize for 3 hours to obtain a decolorized solution, and distill the decolorized solution at normal pressure until the internal temperature reaches 74°C. Collect the distillate in sections, and leave the residual distillate. (4) adding alkali to the distillation residue, adding 40% sodium hydroxide solution, adjusting the pH value of the distillation residue to 9.0 using 40% sodium hydroxide solution, heating and reflux for 3 hours, cooling to 20° C., stirring and crystallizing for 10 hours, and filtering to obtain a crude product of dihydrogen alcohol; 3) Extracting and purifying the crude product of dihydrochloride: (1) Purification of crude bis-aminobutyric acid alcohol: crude bis-aminobutyric acid alcohol is put into a purification tank, and methanol aqueous solution is added for purification to obtain crude bis-aminobutyric acid alcohol purified liquid; 70% methanol aqueous solution of 3 times the mass of phytosterol is added, and the mixture is stirred and heated to 70°C for purification to obtain crude bis-aminobutyric acid alcohol purified liquid; the treatment of crude bis-aminobutyric acid alcohol purified liquid is to adjust the pH value to 9.0 with 40% sodium hydroxide solution, heat and reflux for 3 hours, cool to 20°C, stir and crystallize for 10 hours to obtain crystals and filter, then wash the crystals with drinking water with a pH value of 6.5, stop washing when the outlet pH value reaches the pH value of drinking water, continue filtering until there is no droplet, and dry pump for 3 hours to obtain a first purified bis-aminobutyric acid alcohol. The first purified bis-aminobutyric acid alcohol is purified again according to the above method to obtain a second purified bis-aminobutyric acid alcohol, and the second purified bis-aminobutyric acid alcohol is put into a single cone dryer at a temperature of 100°C and dried for 24 hours, then cooled to room temperature, crushed with a universal grinder and packaged in a fixed amount.

[0019] Example 6 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution: mix 20 g / L glycerol, 8 g / L (NH4)2HPO4, 0.5 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1 g / L corn steep liquor powder (Angel), and 3 g / L defoamer (GPE), add distilled water to make up to 1000 mL, and then sterilize. After sterilization, the pH value is 7.0. Inoculate 2% of the volume of the seed solution with Mycobacterium neoaureum for cultivation for 30 h. When the OD value of the seed solution reaches 12, it is ready for use. (2) Prepare transformation medium: 20 g / L glucose, 8 g / L ammonium citrate tribasic, 0.7 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1.5 g / L Tween80, 3 g / L lecithin (PC40), 3 g / L defoaming agent (GPE), 30 g / L phytosterol (800 mesh). After the above materials are prepared, sterilize them for use. The pH after sterilization is 7.0. (3) Inoculate the qualified seed liquid into the transformation medium at a rate of 15%, and carry out transformation and fermentation for 4 days. When the phytosterol conversion rate reaches ≥95%, stop the fermentation and obtain the phytosterol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The fermentation liquid of the double alcohol fermentation is filtered through a box filter press to obtain a filter cake containing bacteria and products. The filtrate enters the wastewater treatment network as general fermentation wastewater; (2) Extraction: After the filter cake obtained by filtration is added to an extraction tank, methanol 3 to 8 times the mass of the precursor substance phytosterol is added to the extraction tank for extraction, and the extract is obtained by filtration; the extraction is carried out by stirring at room temperature for 1 hour, and then the extraction is carried out by filtering using the filter medium in the extraction tank to obtain an extract. The extraction operation is repeated at least four times until the filter cake is completely extracted, and the multiple extracts are combined; (3) Decolorization of the extract: add activated carbon in an amount 0.05 times the mass of the phytosterols to the extract, raise the temperature to 50°C, stir and decolorize for 1 hour to obtain a decolorized solution, and distill the decolorized solution at normal pressure until the internal temperature reaches 72°C. Collect the distillate in sections, and leave the residual distillate. (4) adding alkali to the distillation residue, adding 40% sodium hydroxide solution, adjusting the pH value of the distillation residue to 8.5 using 40% sodium hydroxide solution, heating and reflux for 1 hour, cooling to 15°C, stirring and crystallizing for 2 hours, and filtering to obtain a crude product of dihydrogen iodide; 3) Extracting and purifying the crude product of dihydrochloride: (1) Purification of crude bis-aminobutyric acid alcohol: crude bis-aminobutyric acid alcohol is put into a purification tank, and methanol aqueous solution is added for purification to obtain crude bis-aminobutyric acid alcohol purified liquid; 40% methanol aqueous solution with a mass of 1.5 times the mass of phytosterol is added, and the mixture is stirred and heated to 60°C for purification to obtain crude bis-aminobutyric acid alcohol purified liquid; the treatment of crude bis-aminobutyric acid alcohol purified liquid is to adjust the pH value to 8.5 with 40% sodium hydroxide solution, heat and reflux for 1 hour, cool to 15°C and stir for crystallization for 2 hours to obtain crystals and filter, and then wash the crystals with drinking water with a pH value of 6.0, stop washing when the outlet pH value reaches the pH value of drinking water, continue filtering until there is no droplet, and dry for 1 hour to obtain a first purified bis-aminobutyric acid alcohol. The first purified bis-aminobutyric acid alcohol is purified again according to the above method to obtain a second purified bis-aminobutyric acid alcohol, and the second purified bis-aminobutyric acid alcohol is put into a single cone dryer at a temperature of 90°C and dried for 12 hours, and then cooled to room temperature, crushed with a universal grinder and packaged in a fixed amount.

[0020] Example 7 A method for extracting and purifying bisphenol A comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution: mix 20 g / L glycerol, 8 g / L (NH4)2HPO4, 0.5 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1 g / L corn steep liquor powder (Angel), and 3 g / L defoamer (GPE), add distilled water to make up to 1000 mL, and then sterilize. After sterilization, the pH value is 7.0. Inoculate 2% of the volume of the seed solution with Mycobacterium neoaureum for cultivation for 30 h. When the OD value of the seed solution reaches 12, it is ready for use. (2) Prepare transformation medium: 20 g / L glucose, 8 g / L ammonium citrate tribasic, 0.7 g / L K2HPO4·3H2O, 3 g / L citric acid, 0.05 g / L ammonium ferric citrate, 0.5 g / L MgSO4·7H2O, 1.5 g / L Tween 80, 3 g / L lecithin (PC40), 3 g / L defoaming agent (GPE), 30 g / L phytosterol (800 mesh). After the above materials are prepared, sterilize them for use. The pH after sterilization is 7.0. (3) Inoculate the qualified seed liquid into the transformation medium at a rate of 15%, and carry out transformation and fermentation for 4 days. When the phytosterol conversion rate reaches ≥95%, stop the fermentation and obtain the phytosterol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The fermentation liquid of the double alcohol fermentation is filtered through a box filter press to obtain a filter cake containing bacteria and products. The filtrate enters the wastewater treatment network as general fermentation wastewater; (2) Extraction: After the filter cake obtained by filtration is added to an extraction tank, methanol in an amount 3 to 8 times the mass of the precursor substance phytosterol is added to the extraction tank for extraction, and the extract is obtained by filtration; the extraction is carried out by stirring at room temperature for 2.5 hours, and then the extraction is carried out by filtering using the filter medium in the extraction tank to obtain an extract. The extraction operation is repeated at least four times until the filter cake is completely extracted, and the extracts of multiple times are combined; (3) Decolorization of the extract: add activated carbon in an amount 1.18 times the mass of the phytosterols to the extract, raise the temperature to 59°C, stir and decolorize for 2.5 hours to obtain a decolorized solution, and distill the decolorized solution at normal pressure until the internal temperature reaches 74°C. Collect the distillate in sections, and leave the residual distillate. (4) adding alkali to the distillation residue, adding 40% sodium hydroxide solution, adjusting the pH value of the distillation residue to 8.8 using 40% sodium hydroxide solution, heating and reflux for 2.5 hours, cooling to 18° C., stirring and crystallizing for 8 hours, and filtering to obtain a crude product of dihydrogen alcohol; 3) Extracting and purifying the crude product of dihydrochloride: (1) Purification of crude bis-aminobutyric acid alcohol: crude bis-aminobutyric acid alcohol is put into a purification tank, and methanol aqueous solution is added for purification to obtain crude bis-aminobutyric acid alcohol purified liquid; 68% methanol aqueous solution of 2.5 times the mass of phytosterol is added, and the mixture is stirred and heated to 65°C for purification to obtain crude bis-aminobutyric acid alcohol purified liquid; the treatment of crude bis-aminobutyric acid alcohol purified liquid is to adjust the pH value to 9.0 with 40% sodium hydroxide solution, heat and reflux for 2.5 hours, cool to 18°C, stir and crystallize for 8 hours to obtain crystals and filter, and then wash the crystals with drinking water with a pH value of 6.3, stop washing when the outlet pH value reaches the pH value of drinking water, continue filtering until there is no droplet, and dry for 2 hours to obtain a first purified bis-aminobutyric acid alcohol. The first purified bis-aminobutyric acid alcohol is purified again according to the above method to obtain a second purified bis-aminobutyric acid alcohol, and the second purified bis-aminobutyric acid alcohol is put into a single cone dryer at a temperature of 95°C and dried for 20 hours, and then cooled to room temperature, crushed with a universal grinder and packaged in a fixed amount.

[0021] The finished products prepared in Example 1, Example 2, Example 3, Example 4, Example 5, Example 6 and Example 7 were tested. Figure 1 As shown, the finished product prepared in the embodiment is bis-decanoic acid, and the bis-decanoic acid prepared in Example 1, Example 2, Example 3, Example 4, Example 5, Example 6, and Example 7 are compared, and the comparison results are shown in Table 1; Table 1 In summary, Examples 1, 2, 3, and 4 did not use sodium hydroxide solution for impurity removal, resulting in a lower content of dihydrogen ethanol. After using sodium hydroxide solution for impurity removal in Examples 5, 6, and 7, the content of dihydrogen ethanol was significantly improved.

[0022] In Example 1, Example 2, Example 3, and Example 4, no sodium hydroxide solution was used for impurity removal, and the moisture content of the final product was 60-70%. In Example 5, Example 6, and Example 7, sodium hydroxide solution was used for impurity removal, and the moisture content of the final product was 30-40%, which effectively improved the drying efficiency of the final product.

[0023] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A method for extracting and purifying bisphenol A, characterized in that: The method for extracting and purifying the double-decanoic acid alcohol comprises the following steps: 1) Fermentation: The fermentation comprises the following steps: (1) Prepare seed solution; (2) Inoculate the seed solution with Mycobacterium neoaureum at a rate of 1%-3% for cultivation, and keep the qualified ones for future use; (2) Prepare transformation medium; (3) Inoculate the qualified seed liquid into the transformation medium at an inoculum rate of 5%-15%, add 30 g / L of phytosterol to the medium, and carry out transformation and fermentation for 3-4 days. When the phytosterol conversion rate is ≥95%, stop the fermentation to obtain the double alcohol fermentation liquid; 2) Extraction of the fermentation broth of the double alcohol reduction: The extraction operation comprises the following steps: (1) Filtration: The filter cake is obtained by hydraulic filtration of the double alcohol fermentation; (2) Extraction: After the filter cake is added to the extraction tank, it is extracted with methanol and filtered to obtain the extract; (3) Decolorization of the extract: After the extract is added to the decolorization tank, activated carbon is added for decolorization and filtration to obtain a decolorized liquid; (4) Distillation of decolorizing liquid: The decolorizing liquid is distilled at normal pressure, and the distillate is collected in sections, and the residual distillate is left; (5) treating the distilled residual liquid with alkali to obtain a crude dihydrochloride; 3) Purification after extraction of the crude product of dihydrochloride: the crude product purification comprises the following steps: (1) Purification of crude dihydrochloride: The crude dihydrochloride is put into a purification tank, and methanol aqueous solution is added for purification to obtain a purified dihydrochloride crude product liquid; (2) Treatment of the purified liquid of crude dihydrochloride: adding alkali to the purified liquid of crude dihydrochloride to adjust the solution, obtain crystals and filter them; (3) Crystallization treatment: washing with drinking water to obtain a purified product of dihydrochloric acid; (4) re-purification: re-purifying the first purified product of dihydrochloride according to the methods of steps (1), (2) and (3) to obtain a second purified product of dihydrochloride; 4) Post-processing: The secondary purified product of dihydrochloride is dried and then crushed and packaged.

2. A method for extracting and purifying double-decanoic acid alcohol according to claim 1, characterized in that: The index of the qualified seed solution in step 1) is that the seed OD value reaches 10-12.

3. A method for extracting and purifying dihydrochloric acid according to claim 1, characterized in that: The extraction operation in step 2) is to add methanol 3 to 8 times the mass of the precursor phytosterol, stir at room temperature for 1 to 3 hours for extraction, and then filter using the filter medium in the extraction tank to obtain an extract.

4. A method for extracting and purifying dihydrochloric acid according to claim 1, characterized in that: The extraction operation in step 2) is repeated at least four times until the filter cake is completely extracted, and the extraction degree of the filter cake is detected by TLC detection method.

5. A method for extracting and purifying dihydrochloric acid according to claim 1, characterized in that: The decolorization operation in step 2) is to add activated carbon in an amount of 0.05 to 0.2 times the mass of the phytosterols to the extract, raise the temperature to 50-60°C, stir and decolorize for 1-3 hours, and distill after the decolorization is completed until the internal temperature is 72-74°C.

6. A method for extracting and purifying dihydrochloric acid according to claim 1, characterized in that: In the step 2), the distilled solvent is recovered and reused, and the solvent with a concentration lower than 40% is separately collected and processed to obtain methanol that can be recovered and reused.

7. A method for extracting and purifying dihydrochloric acid according to claim 1, characterized in that: In the step 2), the distillation residue is added with alkali to adjust the pH value of the distillation residue to 8.5-9.0 using 40% sodium hydroxide solution, the temperature is raised and refluxed for 1-3 hours, the temperature is lowered to 15-20° C. and stirred for crystallization for 2-10 hours, and the crude dihydrochloride is obtained by filtration.

8. A method for extracting and purifying dihydrochloric acid according to claim 1, characterized in that: In the step 3), a 40%-70% methanol aqueous solution with a mass of 1.5 to 3 times the mass of phytosterol is added to purify the crude product of bisulphonic acid, and the mixture is stirred and heated to 60-70°C to purify the crude product to obtain a bisulphonic acid crude product purified solution; the bisulphonic acid crude product purified solution is treated by adjusting the pH value to 8.5-9.0 with a 40% sodium hydroxide solution, heating and reflux for 1-3 hours, cooling to 15-20°C, stirring for crystallization for 2-10 hours, and filtering to obtain a crystal.

9. A method for extracting and purifying dihydrochloric acid according to claim 1, characterized in that: In the step 3), the crystals are treated by washing with drinking water having a pH value of 6.0-6.5, and washing is stopped when the outlet pH value reaches the pH value of drinking water, and filtering is continued until there are no droplets, and dry extraction is performed for 1-3 hours to obtain a purified product of dihydrochloride.

10. The method for extracting and purifying the dihydrochloric acid according to claim 1, characterized in that: In the step 4), the post-treatment is to put the secondary purified product of dihydrogen sulfoxide into a single cone dryer at a temperature of 90-100° C. for 12-24 hours, then cool it to room temperature, crush it with a universal crusher, and then pack it in a fixed amount.

Images