CN105603018B - Method for indirectly preparing D-sodium erythorbate - Google Patents
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Abstract
The invention discloses a method for indirectly preparing D-sodium erythorbate, belonging to the technical field of biochemical separation and synthesis. The invention takes fermentation liquor of Serratia marcescens (Serratia marcocens) SDSPY-136 strain as raw material, and crude sodium D-isoascorbate is obtained by fermentation liquor pretreatment, 2-keto-D-gluconic acid methyl esterification reaction and conversion reaction; and refining and purifying to obtain the D-sodium erythorbate product. The method takes serratia marcescens as a production strain to ferment and obtain the 2-keto-D-calcium gluconate, combines a plurality of impurity removal modes to convert the treatment fluid into the D-sodium erythorbate, and has the advantages of high yield and conversion rate of the obtained fermentation liquor 2-keto-D-calcium gluconate, high purity of the D-sodium erythorbate crude product, easy refining, improved total yield and good popularization and application prospect.
Description
Technical Field
The invention belongs to the technical field of biochemical separation and synthesis, and particularly relates to a method for indirectly preparing D-sodium erythorbate.
Background
D-isoascorbic acid (EA), also known as Erythorbic acid or araboascorbic acid, is an optical isomer of sodium L-ascorbate, abbreviated as eryvc. D-isoascorbic acid and its Sodium salt (Sodium erythorbate, EN) have very strong reducibility, the antiseptic and antioxidant effects are greatly superior to vitamin C, the heat stability is also stronger than vitamin C, and the price is only 1/2 of vitamin C. The isoascorbic acid and the sodium salt thereof are safe and effective food additives and are widely applied to various foods. Sodium isoascorbate is white or yellowish needle crystal, has no odor, slightly salty taste, stability in air, and color darkening when exposed to light, is easily soluble in water, and slightly soluble in methanol and hexanol. The preparation method of the D-sodium erythorbate mainly comprises an indirect fermentation method, a direct fermentation method and an enzyme method. At present, the industrial production adopts an indirect fermentation method, namely, the fermentation and the synthesis are combined for production.
2-keto-D-gluconic acid (2-keto-D-gluconic acid, 2-KGA) is a key intermediate in the industrial production process of a food additive (antioxidant) D-erythorbic acid and sodium salt thereof. The molecular formula of 2-KGA is C6H10O7And has a molecular weight of 194. The 2-KGA is white fine crystal in appearance, slightly salty, odorless, easily soluble in water and slightly acidic in aqueous solution, has optical activity, and has specific optical activity of [ alpha ]]25D = -88 °. The 2-KGA producing strains are widely distributed, and most of the strains are wild types and belong to Pseudomonas (Pseudomonas), Erwinia (Erwinia), Serratia (Serratia) and Acetobacter (Acetobacter).
At present, 2-KGA producing bacteria at home and abroad usually adopt pseudomonas strains, and the yield and purity of D-sodium erythorbate obtained by processing fermentation liquor of the pseudomonas strains through esterification and the like are lower. The method for developing the D-sodium erythorbate with high yield and high purity is significant.
The invention discloses a Chinese invention patent ' CN 104830712A ' for producing high-purity 2-keto-D-gluconic acid Serratia marcescens strain ' and the method adopts Serratia marcescens (Serratia marcocens) SDSPY-136 strain to ferment and produce high-purity 2-keto-D-gluconic acid, the strain is preserved in China general microbiological culture Collection center (CGMCC), the preservation number is N0.10548, the preservation time is: year 2015, 2 months and 9 days.
Disclosure of Invention
In order to make up the defects of the prior art, the invention provides a method for preparing D-isoascorbic acid by using fermentation liquor of Serratia marcescens (SDSPY-136) strain which can produce high-purity 2-keto-D-gluconic acid.
The technical scheme of the invention is as follows:
a method for indirectly preparing D-sodium erythorbate comprises taking fermentation liquor of Serratia marcescens (Serratia marcescens) SDSPY-136 strain as raw material, pretreating the fermentation liquor, performing 2-keto-D-methyl gluconate esterification reaction, and converting to obtain crude sodium D-erythorbate; refining and purifying to obtain a D-sodium erythorbate product; the Serratia marcescens (Serratia marcescens) SDSPY-136 strain is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, and the preservation number is CGMCC No: 10548, the preservation time is: year 2015, 2 months and 9 days.
As a preferred scheme, the preparation method for indirectly preparing the D-sodium erythorbate comprises the following specific steps:
1) fermentation of bacterial species
Inoculating the seed liquid of the Serratia marcescens (Serratia marcescens) SDSPY-136 strain into a fermentation culture medium according to the inoculation amount of 8-12%, and performing fermentation culture for 30-40h under the conditions of 29-31 ℃ and 400-600r/min to obtain 2-keto-D-gluconic acid fermentation liquid;
2) pretreatment of fermentation liquor
Adding concentrated sulfuric acid into the clear solution obtained after the centrifugation of the 2-keto-D-gluconic acid fermentation liquor, then adding active carbon into the acidified clear solution under the water bath condition of 50-95 ℃, and stirring and decoloring; filtering to remove active carbon after decolorization, passing the obtained filtrate through a cation exchange resin column, removing impurities by the cation exchange resin column, and distilling the collected treated liquid under reduced pressure to obtain a 2-keto-D-gluconic acid concentrated solution with the solid content of 70-90%;
3) methyl esterification reaction
Adding methanol into the 2-keto-D-gluconic acid concentrated solution, reacting for 5-10h at 60-100 ℃ by using sulfuric acid as a catalyst, concentrating after the reaction is finished, crystallizing at 4 ℃, filtering, and drying to obtain 2-keto-D-gluconic acid methyl ester salt;
4) conversion reaction
Dissolving the 2-keto-D-methyl gluconate in methanol, adding methanol-sodium hydroxide mixed solution, reacting at 55-70 deg.C for 13-20min, crystallizing at 4 deg.C, and filtering to obtain D-isoascorbic acid crude sodium salt;
5) refining of D-sodium erythorbate
Dissolving the obtained D-isoascorbic acid crude sodium salt in water, adding active carbon and oxalic acid, heating to 50-70 ℃, keeping the temperature and stirring for 3-8min, and then carrying out suction filtration while the solution is hot to obtain a filtrate; and cooling, crystallizing, filtering, washing with 90-100% ethanol, and drying to obtain D-sodium erythorbate product.
Preferably, the seed culture medium used for obtaining the seed liquid in the step 1) comprises 5g/L of peptone, 5g/L of sodium chloride and 3g/L of beef extract, and the pH value is 7.0; the fermentation medium comprises 180g/L of glucose, 9.01 g/L of corn steep liquor, 1 g/L of potassium dihydrogen sulfate, 0.4 g/L of magnesium sulfate, 0.037 g/L of manganese chloride, 0.036 g/L of ferrous sulfate and 65 g/L of calcium carbonate.
Preferably, the adding amount of the activated carbon in the step 2) is 5-15% of the solid content, and the decoloring time is at least 0.5 h.
Preferably, in the step 2), the cation exchange resin column is a 732# cation exchange resin column, the height-diameter ratio of the cation exchange resin column is 9:1-10:1, and the flow rate is 1-1.1 times of the column volume per hour.
Preferably, the adding amount of the methanol in the step 3) meets the following requirements: adding 5-1.5mL of methanol into each gram of 2-keto-D-gluconic acid concentrated solution.
Preferably, in the step 4), the adding amount of the methanol in the 2-keto-D-gluconic acid methyl ester salt satisfies the following conditions: adding 10-10.5mL of methanol into each gram of 2-keto-D-gluconic acid methyl ester salt; the addition amount of the methanol-sodium hydroxide mixed solution meets the following requirements: adding 4-4.5mL of methanol-sodium hydroxide mixed solution into each gram of 2-keto-D-gluconic acid methyl ester salt.
Preferably, in the step 5), the adding amount of the activated carbon and the oxalic acid is 0.5-0.7% of the solid content.
The invention has the beneficial effects that:
the method takes serratia marcescens as a production strain to ferment and obtain the 2-keto-D-calcium gluconate, combines a plurality of impurity removal modes to convert the treatment fluid into the D-sodium erythorbate, and has the advantages of high yield and conversion rate of the obtained fermentation liquor 2-keto-D-calcium gluconate, high purity of the D-sodium erythorbate crude product, easy refining, improved total yield and good popularization and application prospect.
Drawings
The invention will be further described with reference to the accompanying drawings.
FIG. 1 is a sample of sodium D-erythorbate obtained in example 1 of the present invention;
FIG. 2 is a liquid chromatogram of a D-sodium erythorbate sample obtained in example 1 of the present invention;
FIG. 3 is a liquid chromatogram of a standard D-sodium erythorbate sample.
Serratia marcescens (Serratia marcocens) SDSPY-136 strain, which is preserved in China general microbiological culture Collection center (CGMCC), and the preservation address is as follows: the Xilu No. 1 Hospital, Chaozhou, Chaoyang, Beijing, has a preservation number of N0.10548, and a preservation time: year 2015, 2 months and 9 days.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited thereto.
The seed culture medium comprises the following components: 5g/L of peptone, 5g/L of sodium chloride and 3g/L of beef extract, and the pH value is 7.0.
The fermentation medium comprises the following components: 180g/L glucose, 9.01 g/L corn steep liquor, 1 g/L potassium dihydrogen sulfate, 0.4 g/L magnesium sulfate, 0.037 g/L manganese chloride, 0.036 g/L ferrous sulfate and 65 g/L calcium carbonate.
Example 1
The preparation method for indirectly preparing the D-sodium erythorbate comprises the following specific steps:
1) fermentation of bacterial species
The strain of Serratia marcescens (Serratia marcescens) CGMCC N0.10548 is grafted to a slant for activation.
Preparing a seed culture medium according to the composition of the liquid seed culture medium, wherein the liquid loading capacity of a 500ml triangular flask is 50ml, steam sterilization is carried out for 20 minutes at the temperature of 121 ℃, cooling to room temperature is carried out, 2 rings of slant strains are inoculated, the slant strains are placed on a shaking bed with the rotating speed of 200r/min and the rotating radius of 40mm, culture is carried out for 12 hours at the temperature of 30 ℃, a seed solution is obtained, and the light absorption value of the seed solution is measured to be 0.262 at the wavelength of 600 nm;
preparing fermentation medium with a 5L fermentation tank according to 3L liquid loading amount, adjusting pH to 7.0, sterilizing at 115 deg.C for 20min, cooling with circulating water to 30 deg.C, inoculating 300 ml of cultured seed solution, immediately sampling after inoculation, and determining glucose concentration to be 185 g/L.
The initial fermentation tank parameters are set as that the stirring speed is 350r/min, and the ventilation rate is 0.7 ~ 0.8.8 Nm3H, the pressure in the tank is 0.065 MPa. The fermentation temperature is controlled to be 30 +/-0.2 ℃ in the fermentation process, and the dissolved oxygen is controlled to be 35% by adjusting the stirring speed and the ventilation quantity.
Sampling every 4 hours in the fermentation process, measuring the contents of glucose and 2-keto-D-gluconic acid in the fermentation broth and the concentration of thalli, when the content of the glucose is lower than 10 g/L, sampling every 2 hours to measure the contents of the glucose and the 2-keto-D-gluconic acid in the fermentation broth, ending the fermentation when the content of the 2-keto-D-gluconic acid in the fermentation broth is not increased, wherein the fermentation period is 36 hours.
And after the fermentation is finished, centrifuging the fermentation liquor for 10 minutes at 4000r/min, taking supernate to determine that the contents of glucose and 2-keto-D-gluconic acid are respectively 0.02g/L and 188.9g/L, wherein the conversion rate of converting the glucose into the 2-keto-D-gluconic acid is 102.11%.
2) Pretreatment of fermentation broths
Determination of Ca after centrifugation of fermentation broth2+The content of the Ca is 0.66mol/L, and 0.66mol/L concentrated sulfuric acid is added to lead the Ca to be2+The precipitate is converted into calcium sulfate, and 2.7L of acidified clear liquid is obtained after centrifugation; 19.2 percent of solid content, adding active carbon accounting for 5 percent of the solid content under the condition of 80 ℃ water bath, continuously stirring, decoloring for 0.5h, and filtering after decoloring; the filtrate passes through 732# cation exchange resin, and 3.1L of effluent liquid is obtained; the treated liquid collected by removing impurities from the column was concentrated at 55 ℃ under reduced pressure until the solid content became 80%, and the total amount was 501.3 g.
3) Methyl esterification reaction
Taking 100g of 2-keto-D-gluconic acid concentrated solution, adding 500ml of methanol, taking concentrated sulfuric acid as a catalyst, reacting for 7 hours in a water bath at 80 ℃, and supplementing methanol into a reactor. Concentrating, standing at 4 deg.C, vacuum filtering, washing with methanol, and mixing the washing solution with the clear liquid. The precipitate was dried at 40 ℃ to obtain 64.12g of methyl ester salt.
4) Conversion reaction
Adding 60g of methyl salt into 600ml of methanol solution, stirring to completely dissolve the methyl salt, adding 250ml of methanol-NaOH solution, reacting in a water bath at 65 ℃ for 17min, standing at 4 ℃ for a period of time, and performing suction filtration to obtain 50.82g of crude sodium salt with the purity of 98.1%. Wherein the mass ratio of methanol to sodium hydroxide in the methanol-NaOH solution is 10: 1.
5) Refining of D-sodium erythorbate
Dissolving 50.5g of crude sodium salt in water, adding 0.3g of oxalic acid and 0.3g of activated carbon, keeping the temperature of the mixture in a water bath to 60 ℃, keeping the temperature and stirring the mixture for 5min, and filtering the mixture while the mixture is hot to obtain 150ml of filtrate. Cooling, crystallizing, filtering, washing crystal with 95% ethanol, drying at 50 deg.C to obtain 48.82g of D-sodium erythorbate product with yield of 95.6% and purity of 99.5%. Liquid phase analysis of the product shows that the product is sodium erythorbate.
Example 2
The preparation method for indirectly preparing the D-sodium erythorbate comprises the following specific steps:
1) fermentation of bacterial species
The strain of Serratia marcescens (Serratia marcescens) CGMCC N0.10548 is grafted to a slant for activation.
Preparing a seed culture medium according to the composition of the liquid seed culture medium, wherein the liquid loading capacity of a 500ml triangular flask is 50ml, steam sterilization is carried out for 20 minutes at the temperature of 121 ℃, cooling to room temperature is carried out, 2 rings of slant strains are inoculated, the slant strains are placed on a shaking bed with the rotating speed of 200r/min and the rotating radius of 40mm, culture is carried out for 12 hours at the temperature of 30 ℃, a seed solution is obtained, and the light absorption value of the seed solution is measured to be 0.265 at the wavelength of 600 nm;
preparing a fermentation culture medium by a 10L fermentation tank according to the liquid loading amount of 7L, adjusting the pH value to 7.0, sterilizing the solid tank at 115 ℃ for 20 minutes, cooling the solid tank to 30 ℃ by circulating water, inoculating 700 ml of cultured seed liquid, immediately sampling after inoculation, and determining the glucose concentration to be 180 g/L.
The initial fermentation tank parameters are set as that the stirring speed is 350r/min, and the ventilation rate is 0.8 ~ 0.9.9 Nm3H, the pressure in the tank is 0.065 MPa. The fermentation temperature is controlled to be 30 +/-0.2 ℃ in the fermentation process, and the dissolved oxygen is controlled to be 35% by adjusting the stirring speed and the ventilation quantity.
Sampling every 4 hours in the fermentation process, measuring the contents of glucose and 2-keto-D-gluconic acid in the fermentation broth and the concentration of thalli, when the content of the glucose is lower than 10 g/L, sampling every 2 hours to measure the contents of the glucose and the 2-keto-D-gluconic acid in the fermentation broth, ending the fermentation when the content of the 2-keto-D-gluconic acid in the fermentation broth is not increased, wherein the fermentation period is 35 hours.
And after the fermentation is finished, centrifuging the fermentation liquor for 10 minutes at 4000r/min, taking supernate to determine that the contents of glucose and 2-keto-D-gluconic acid are respectively 0.03g/L and 183.12g/L, wherein the conversion rate of converting the glucose into the 2-keto-D-gluconic acid is 101.73%.
2) Pretreatment of fermentation broths
Determination of Ca after centrifugation of fermentation broth2+The content of the Ca is 0.67mol/L, and 0.67mol/L concentrated sulfuric acid is added to lead the Ca to be2+The precipitate is converted into calcium sulfate, and 6.5L of acidified clear liquid is obtained after centrifugation; 19.5 percent of solid content, adding active carbon accounting for 5 percent of the solid content under the condition of 80 ℃ water bath, continuously stirring, decoloring for 0.5h, and filtering after decoloring; passing the filtrate through 732# cation exchange resin column, and discharging 7.2L of liquid; the treated liquid collected by removing impurities from the column was concentrated at 55 ℃ under reduced pressure until the solid content was 79.5%, and 1.1kg in total.
3) Methyl esterification reaction
Taking 100g of 2-keto-D-gluconic acid concentrated solution, adding 500ml of methanol, taking concentrated sulfuric acid as a catalyst, reacting for 7 hours in a water bath at 80 ℃, and supplementing methanol into a reactor. Concentrating, standing at 4 deg.C, vacuum filtering, washing with methanol, and mixing the washing solution with the clear liquid. The precipitate was dried at 40 ℃ to obtain 64.10g of methyl ester salt.
4) Conversion reaction
Adding 60g of methyl salt into 600ml of methanol solution, stirring to completely dissolve the methyl salt, adding 250ml of methanol-NaOH solution, reacting in a water bath at 65 ℃ for 17min, standing at 4 ℃ for a period of time, and performing suction filtration to obtain 50.93g of crude sodium salt with the purity of 98.0%. Wherein the mass ratio of methanol to sodium hydroxide in the methanol-NaOH solution is 9: 1.
5) Refining of D-sodium erythorbate
Dissolving 50.0g of crude sodium salt in water, adding 0.3g of oxalic acid and 0.3g of activated carbon, keeping the temperature of the mixture in a water bath to 60 ℃, keeping the temperature and stirring the mixture for 5min, and filtering the mixture while the mixture is hot to obtain 155ml of filtrate. Cooling, crystallizing, filtering, washing crystal with 95% ethanol, drying at 50 deg.C to obtain 48.79g of D-sodium erythorbate product with yield of 96.1% and purity of 99.6%. Liquid phase analysis of the product shows that the product is sodium erythorbate.
Comparative example 1
Serratia marcescens (a), (b), (c), (d) and (d)Serratia marcescensSDSPY-136) was replaced with a fermentation broth of a strain of pseudomonas, the remainder being the same as in example 1.
The comparative example D-sodium erythorbate gave a yield of 90.2 and a purity of 93.3.
Comparative example 2
The procedure of "passing the filtrate through No. 732 cation exchange resin column" in step 2) was omitted, and the filtrate was directly concentrated, as in example 1.
The comparative example D-sodium erythorbate gave a yield of 85.2% and a purity of 90.1%.
Comparative example 3
In the step 5), only 0.3g of activated carbon is added after the D-isoascorbic acid crude sodium salt is dissolved in water, and no oxalic acid is added; the rest is the same as in example 1.
The comparative example D-sodium erythorbate gave an yield of 87.3% and a purity of 91.8%.
The glucose was measured as follows:
the measurement was carried out using a SBA-4D type membrane bioreactor manufactured by institute of biology, institute of sciences, Shandong province. The determination principle is that the immobilized glucose dehydrogenase film is used for specifically determining the glucose content.
The 2-keto-D-gluconic acid was measured by the following method:
iodometry was used. The determination principle is as follows: the 2-keto-D-gluconic acid molecule contains carboxyl and alcoholic hydroxyl which can directly generate esterification reaction and carbonyl which can generate tautomerization reaction, and the 2-keto-D-gluconic acid aqueous solution can be quantitatively converted into D-erythorbic acid by heating under the strong acid condition. Using D-erythorbic acid with I2The redox reaction of (2) can quantitatively determine the mass concentration of D-erythorbic acid, and then convert the mass concentration into the mass concentration of 2-keto-D-gluconic acid.
The conversion of 2-keto-D-gluconic acid was measured by the following method:
conversion ratio (%) of 2-keto-D-gluconic acid (= 2-keto-D-gluconic acid production (g/L)/[ glucose content in initial medium (g/L) — glucose content in post-fermentation medium (g/L) ]
The above Ca2+The measurement was carried out as follows: EDTA titration method for measuring calcium in water by GB/T7476-1987
The above D-sodium erythorbate was measured by the following method: GB8273-2008 determination of food additive D-sodium erythorbate.
The identification of the sodium D-erythorbate is carried out according to the following method: high performance liquid chromatography. The chromatographic conditions are as follows: an UltiMate 3000 series VWD-3000 variable wavelength detector; the column was a Prontosil 1202102C 18 column (10 μm, 4.6mm i.d.. times.250 mm); the mobile phase is prepared into 0.1mol/L KH by secondary distilled water2PO4Adjusting the pH value to 2.5 by phosphoric acid; the flow rate is 0.5 mL/min; the injection volume is 20 mu L; the detection wavelength is 210 nm; the column temperature was 30 ℃.
Claims (5)
1. A preparation method for indirectly preparing D-sodium erythorbate is characterized by comprising the following steps: taking fermentation liquor of Serratia marcescens (SDSPY-136) strain as a raw material, and performing pretreatment, 2-keto-D-gluconic acid methyl esterification reaction and conversion reaction on the fermentation liquor to obtain D-isoascorbic acid crude sodium salt; refining and purifying to obtain a D-sodium erythorbate product; the Serratia marcescens (Serratia marcescens) SDSPY-136 strain is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, and the preservation number is CGMCC No: 10548, the preservation time is: 2015, 2 months and 9 days;
the preparation method for indirectly preparing the D-sodium erythorbate comprises the following specific steps:
1) fermentation of bacterial species
Inoculating the seed liquid of the Serratia marcescens (Serratia marcescens) SDSPY-136 strain into a fermentation culture medium according to the inoculation amount of 8-12%, and performing fermentation culture for 30-40h under the conditions of 29-31 ℃ and 400-600r/min to obtain 2-keto-D-gluconic acid fermentation liquid;
the seed culture medium used for obtaining the seed liquid in the step 1) comprises 5g/L of peptone, 5g/L of sodium chloride and 3g/L of beef extract, and the pH value is 7.0; the fermentation medium comprises 180g/L of glucose, 9.01 g/L of corn steep liquor, 1 g/L of potassium dihydrogen sulfate, 0.4 g/L of magnesium sulfate, 0.037 g/L of manganese chloride, 0.036 g/L of ferrous sulfate and 65 g/L of calcium carbonate;
2) pretreatment of fermentation liquor
Adding concentrated sulfuric acid into the clear solution obtained after the centrifugation of the 2-keto-D-gluconic acid fermentation liquor, then adding active carbon into the acidified clear solution under the water bath condition of 50-95 ℃, and stirring and decoloring; filtering to remove active carbon after decolorization, passing the obtained filtrate through a cation exchange resin column, removing impurities by the cation exchange resin column, and distilling the collected treated liquid under reduced pressure to obtain a 2-keto-D-gluconic acid concentrated solution with the solid content of 70-90%; the cation exchange resin column is No. 732 cation exchange resin column, the height-diameter ratio of the cation exchange resin column is 9:1-10:1, and the flow rate is 1-1.1 times of the column volume per hour;
3) methyl esterification reaction
Adding methanol into the 2-keto-D-gluconic acid concentrated solution, reacting for 5-10h at 60-100 ℃ by using sulfuric acid as a catalyst, concentrating after the reaction is finished, crystallizing at 4 ℃, filtering, and drying to obtain 2-keto-D-gluconic acid methyl ester salt;
4) conversion reaction
Dissolving the 2-keto-D-methyl gluconate in methanol, adding methanol-sodium hydroxide mixed solution, reacting at 55-70 deg.C for 13-20min, crystallizing at 4 deg.C, and filtering to obtain D-isoascorbic acid crude sodium salt;
5) refining of D-sodium erythorbate
Dissolving the obtained D-isoascorbic acid crude sodium salt in water, adding active carbon and oxalic acid, heating to 50-70 ℃, keeping the temperature and stirring for 3-8min, and then carrying out suction filtration while the solution is hot to obtain a filtrate; and cooling, crystallizing, filtering, washing with 90-100% ethanol, and drying to obtain D-sodium erythorbate product.
2. The process for indirectly preparing sodium D-erythorbate according to claim 1, wherein: in the step 2), the adding amount of the activated carbon is 5-15% of the solid content, and the decoloring time is at least 0.5 h.
3. The method for indirectly preparing sodium D-erythorbate according to claim 1, wherein the amount of methanol added in step 3) is such that: adding 5-1.5mL of methanol into each gram of 2-keto-D-gluconic acid concentrated solution.
4. The process for indirectly preparing sodium D-erythorbate according to claim 1, wherein in step 4), the amount of methanol added to the methyl 2-keto-D-gluconate salt is such that: adding 10-10.5mL of methanol into each gram of 2-keto-D-gluconic acid methyl ester salt; the addition amount of the methanol-sodium hydroxide mixed solution meets the following requirements: adding 4-4.5mL of methanol-sodium hydroxide mixed solution into each gram of 2-keto-D-gluconic acid methyl ester salt.
5. The method for indirectly preparing sodium D-erythorbate according to claim 1, wherein in step 5), the amount of the activated carbon and the oxalic acid added is 0.5% -0.7% of the solid content.
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| CN108129427A (en) * | 2017-12-29 | 2018-06-08 | 郑州拓洋生物工程有限公司 | ISOASCORBIC ACID potassium and its preparation method and application |
| CN110294726A (en) * | 2019-07-29 | 2019-10-01 | 新拓洋生物工程股份有限公司 | The preparation method of ISOASCORBIC ACID potassium |
| CN110642817A (en) * | 2019-10-22 | 2020-01-03 | 新拓洋生物工程股份有限公司 | Isovitamin C potassium refining method |
| CN111337613B (en) * | 2020-04-18 | 2023-03-21 | 新拓洋生物工程有限公司 | High performance liquid detection method for D-isoascorbic acid potassium |
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