CN107382760B - Separation and purification method of levodopa - Google Patents
Separation and purification method of levodopa Download PDFInfo
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- CN107382760B CN107382760B CN201710711008.2A CN201710711008A CN107382760B CN 107382760 B CN107382760 B CN 107382760B CN 201710711008 A CN201710711008 A CN 201710711008A CN 107382760 B CN107382760 B CN 107382760B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/38—Separation; Purification; Stabilisation; Use of additives
- C07C227/44—Stabilisation; Use of additives
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/38—Separation; Purification; Stabilisation; Use of additives
- C07C227/40—Separation; Purification
- C07C227/42—Crystallisation
Abstract
The invention provides a separation and purification method of levodopa, which comprises the steps of firstly adjusting a solution to be alkaline under the action of an ascorbic acid antioxidant, dissolving the levodopa, and then adjusting the solution to be acidic, so that the levodopa is crystallized and separated out. The method uses the ascorbic acid as the antioxidant in the reaction process, so that the levodopa is prevented from being oxidized in the separation and purification process, and the yield of the levodopa is improved; according to the invention, the levodopa is precipitated by using acetic acid, so that the condition that the acidity is too strong to influence the growth speed of the levodopa precipitate is avoided, the growth speed of the levodopa precipitate is controlled, and the precipitation amount of the levodopa precipitate is increased; and the introduction of acidic substances with oxidability can be avoided, and the possibility of oxidation in the separation and purification process of levodopa is reduced, so that the yield of levodopa is improved. The results of the examples show that the product yield of the method provided by the invention can reach 88-91%, the purity is more than 99.8%, and the method meets the requirements of CP/USP/EP.
Description
Technical Field
The invention relates to the field of medicinal chemistry, in particular to a separation and purification method of levodopa.
Background
Levodopa, also known as 3, 4-dihydroxyphenylalanine, is an important bioactive substance. Levodopa is a precursor of neurotransmitter dopamine, can enter brain circulation through a blood brain barrier to reach a central nervous system, and is converted into dopamine under the action of central nerve decarboxylase, so that the content of the dopamine in brain tissues is increased, and the Parkinson disease is treated. Besides, levodopa has the effect of treating amblyopia and heart failure, and although there are some side effects after long-term use, no better alternative drugs are available at present over the past forty years.
According to statistics, the total global sales of levodopa reaches $ 3.3 billion in 2015, the total yield reaches more than 2000 tons in 97 th-ranking worldwide popular drug, and the total global sales is expected to reach $ 10 billion in 5-10 years in the future. In the prior art, the levodopa extraction method is generally obtained by performing two-step crystallization on a conversion solution, or performing membrane filtration on the conversion solution, adding hydrochloric acid for dissolution, and then performing crystallization. However, the yield of levodopa products obtained by the two methods is low.
Disclosure of Invention
In view of this, the present invention aims to provide a method for separating and purifying levodopa to increase the yield of levodopa.
The invention provides a separation and purification method of levodopa, which comprises the following steps:
1) mixing a raw material solution containing levodopa with ascorbic acid to obtain a first mixed solution;
2) adjusting the pH value of the first mixed solution obtained in the step 1) to be alkaline to obtain an alkaline mixed solution;
3) carrying out solid-liquid separation on the alkaline mixed solution obtained in the step 2) to obtain a supernatant;
4) mixing the supernatant obtained in the step 3) with ascorbic acid to obtain a second mixed solution;
5) adjusting the pH value of the second mixed solution obtained in the step 4) to 3-5 by adopting acetic acid, and precipitating levodopa.
Preferably, the concentration of levodopa in the raw material liquid in the step 1) is not lower than 30 g/L.
Preferably, the concentration of the ascorbic acid in the first mixed solution in the step 1) and the concentration of the ascorbic acid in the second mixed solution in the step 4) are 0.5-2 g/L independently.
Preferably, the pH value of the alkaline mixed solution in the step 2) is 10-13.
Preferably, the pH value adjustment in the step 5) sequentially comprises a first acidity adjustment and a second acidity adjustment;
the first acidity adjustment comprises: adjusting the pH value of the second mixed solution obtained in the step 4) to 6-7 by adopting first acetic acid;
the second acidity adjustment includes: and adjusting the pH value of the mixed solution after the first acidity adjustment to 3-5 by adopting second acetic acid.
Preferably, the step 5) further comprises, after pH adjustment: and crystallizing the adjusted acidic mixed solution.
Preferably, the temperature of the crystallization treatment is 2-7 ℃.
Preferably, the time of the crystallization treatment is 2 to 3 hours.
Preferably, the method further comprises the following steps before mixing the supernatant obtained in step 3) with ascorbic acid: and (3) decoloring the supernatant by using activated carbon.
Preferably, after the levodopa is precipitated in the step 5), the solid-liquid separation is performed on the precipitated feed liquid, and the obtained solid substance is the levodopa.
The invention provides a separation and purification method of levodopa, which comprises the steps of firstly adjusting a solution to be alkaline under the action of an ascorbic acid antioxidant, dissolving the levodopa, and then adjusting the solution to be acidic, so that the levodopa is crystallized and separated out. The method uses the ascorbic acid as the antioxidant in the reaction process, so that the levodopa is prevented from being oxidized in the separation and purification process, and the yield of the levodopa is improved; and then the mixed solution of the levodopa and the ascorbic acid is adjusted to be alkaline, so that the levodopa can be dissolved to the maximum extent, and the yield of the levodopa can be improved. According to the invention, the levodopa is precipitated by using acetic acid, so that the condition that the acidity is too strong and the growth speed of the levodopa precipitate is influenced is avoided, the growth speed of the levodopa precipitate is controlled, and the precipitation amount of the levodopa precipitate is increased; and the introduction of acidic substances with oxidability can be avoided, so that the possibility of oxidation in the separation and purification process of the levodopa is reduced, and the yield of the levodopa is further improved. The results of the examples show that the product yield of the method provided by the invention can reach 88-91%, the purity is more than 99.8%, and the method meets the requirements of CP/USP/EP.
Drawings
FIG. 1 is a flow chart of the separation and purification method provided by the present invention.
Detailed Description
The invention provides a separation and purification method of levodopa, which comprises the following steps:
1) mixing a raw material solution containing levodopa with ascorbic acid to obtain a first mixed solution;
2) adjusting the pH value of the first mixed solution obtained in the step 1) to be alkaline to obtain an alkaline mixed solution;
3) carrying out solid-liquid separation on the alkaline mixed solution obtained in the step 2) to obtain a supernatant;
4) mixing the supernatant obtained in the step 3) with ascorbic acid to obtain a second mixed solution;
5) adjusting the pH value of the second mixed solution obtained in the step 4) to 3-5 by adopting acetic acid, and precipitating levodopa.
The levodopa has a three-dimensional structure shown in formula I:
the method comprises the step of mixing raw material liquid containing levodopa with ascorbic acid to obtain a first mixed solution.
In the present invention, the concentration of levodopa in the raw material liquid containing levodopa is not less than 30g/L, and more preferably 30 to 200 g/L. The source of the raw material liquid containing levodopa is not particularly required in the present invention, and the raw material liquid for separation and purification known to those skilled in the art can be used. In the embodiment of the invention, the raw material liquid containing levodopa is specifically a conversion liquid obtained by biocatalysis of a substrate by using a tyrosine phenol lyase; the substrate includes catechol, pyruvate, and ammonia. The invention has no special requirements on the biological catalysis mode of the tyrosine phenol lyase, and the method is well known by the technical personnel in the field.
In the present invention, the biocatalysis of a substrate by the tyrosol lyase preferably comprises the following steps:
(1) providing a substrate solution, wherein the substrate solution comprises, by mass, 8-12 parts of catechol, 10-14 parts of sodium pyruvate, 40-50 parts of ammonium acetate, 1-3 parts of sodium sulfite, 0.5-2 parts of ethylenediamine tetraacetic acid, 0.05-0.2 part of pyridoxal phosphate and 700-800 parts of water;
(2) adjusting the pH value of the substrate solution to 8.15-8.2 by adopting ammonia water to obtain an alkaline substrate solution;
(3) and carrying out a biocatalytic reaction on the obtained alkaline substrate solution by adopting a tyrosine phenol lyase solution to obtain a raw material solution containing levodopa.
In the present invention, the substrate solution is preferably obtained by mixing catechol, sodium pyruvate, ammonium acetate, sodium sulfite, ethylenediaminetetraacetic acid, and pyridoxal phosphate in water; the mixing temperature is preferably 20-22 ℃.
In the invention, the mass ratio of the volume of the tyrosine phenol lyase solution to the catechol is preferably 95-105 mL: 8-12 g; the mass concentration of the tyrosine phenol lyase solution is 5-10%. In the present invention, the biocatalytic reaction is preferably carried out under a nitrogen atmosphere; the temperature of the biological catalytic reaction is preferably 15-18 ℃; the biocatalytic reaction is preferably carried out under the condition of stirring, and the stirring speed is preferably 200-230 rpm.
The invention has no special requirement on the time of the biocatalytic reaction, and the condition is that the levodopa is not generated in the mixed solution any more.
The invention preferably determines whether levodopa is generated in the mixed solution by detecting the residual quantity of the substrate. Specifically, the invention samples and measures the residual quantity of catechol and sodium pyruvate every hour, and when the residual quantity of catechol and sodium pyruvate is not reduced any more, the levodopa is not generated in the mixed solution.
The invention preferably adopts an HPLC detection method to detect the contents of the catechol and the sodium pyruvate.
Sampling every hour to measure the residual quantity of catechol and sodium pyruvate, and adding catechol when the concentration of the catechol is lower than 5-7 g/L to maintain the concentration of the catechol at 5-7 g/L; and when the concentration of the sodium pyruvate is lower than 7-9 g/L, adding the sodium pyruvate to maintain the concentration of the sodium pyruvate at 7-9 g/L. Thus ensuring that the substrate is fully converted into the levodopa under the action of the tyrosine phenol lyase.
After the biocatalytic reaction is completed, the mixed solution obtained by the biocatalytic reaction is preferably mixed with a sulfuric acid solution to obtain a raw material solution containing levodopa. The mass concentration of the sulfuric acid solution is preferably 50% -55%, and the mass ratio of the volume of the sulfuric acid solution to the catechol in the substrate solution is preferably 20-30 mL: 8-12 g.
In the present invention, the raw material solution containing levodopa may be derived from another solution having a levodopa concentration of 30g/L or more.
The present invention does not require any special mixing method for the raw material solution containing levodopa and ascorbic acid, and a solution mixing method known to those skilled in the art can be used.
In the present invention, the concentration of the ascorbic acid in the first mixed solution is preferably 0.5 to 2g/L, and more preferably 1 to 1.5 g/L.
After the first mixed solution is obtained, the pH value of the first mixed solution is adjusted to be alkaline, and an alkaline mixed solution is obtained. According to the invention, strong alkali is preferably adopted to adjust the pH value of the first mixed solution; the strong base is preferably sodium hydroxide and/or potassium hydroxide. In the present invention, the strong base preferably adjusts the pH of the first mixed solution in the form of a strong base solution; the mass-volume concentration of the strong alkali solution is preferably 300-500 g/L.
In the present invention, the adjustment of the pH value is preferably to add the strong alkali solution to the first mixed solution; the addition rate of the strong alkali solution is preferably: 20-40 mL/min.
The invention has no special requirement on the using amount of the strong base so as to realize the adjustment of the pH value of the first mixed solution and obtain the alkaline mixed solution. In the present invention, the pH value of the alkaline mixed solution is preferably 10 to 13, and more preferably 11 to 12. According to the invention, the pH value of the first mixed solution is adjusted to a target range, so that levodopa can be dissolved to the maximum extent, and the yield of levodopa can be improved.
In the present invention, the adjustment of the pH is preferably performed under stirring conditions; the stirring speed is preferably 300-500 rpm.
After the alkaline mixed solution is obtained, the invention carries out solid-liquid separation on the alkaline mixed solution to obtain the supernatant. In the present invention, the solid-liquid separation is preferably centrifugal separation; the temperature of the centrifugal separation is preferably 0-8 ℃, and further preferably 3-5 ℃; the rotation speed of the centrifugal separation is preferably 3000-5000 rpm, and more preferably 3500-4500 rpm; the time for centrifugal separation is preferably 10-60 min, and more preferably 20-50 min.
After the supernatant is obtained, the present invention mixes the supernatant with ascorbic acid to obtain a second mixed solution.
In the invention, the concentration of the ascorbic acid in the second mixed solution is preferably 0.5-2 g/L.
In the present invention, the mixing manner of the supernatant and the ascorbic acid is not particularly limited, and the supernatant and the ascorbic acid may be sufficiently mixed.
In the present invention, the supernatant and the ascorbic acid preferably further include, before mixing: and (3) decoloring the supernatant by using activated carbon. In the present invention, the decoloring treatment is preferably performed by mixing the supernatant with activated carbon. In the invention, the volume ratio of the mass of the activated carbon to the supernatant is preferably 0.02-0.04 g: 1L of the compound. In the present invention, the mixing is preferably performed under stirring conditions; the stirring speed is preferably 100 to 200rpm, and more preferably 150 to 180 rpm. The invention realizes the adsorption of the color in the supernatant by the activated carbon by carrying out the decoloring treatment in the mixing process of the supernatant and the activated carbon. In the invention, the temperature of the decolorization treatment is preferably 30-50 ℃, and more preferably 35-45 ℃; the time of the decoloring treatment is preferably 2 to 3 hours.
After the decolorization treatment, the present invention preferably removes activated carbon. The specific implementation mode of the activated carbon removal method is not particularly required, and the solid removal method in the solution is well known to those skilled in the art. The invention preferably adopts a centrifugal separation or filtration mode to remove the activated carbon.
After the second mixed solution is obtained, the pH value of the second mixed solution is adjusted to 3-5 by adopting acetic acid, and levodopa is separated out. In the process of adjusting the pH value to 3-5, the levodopa is separated out from the solution. The invention has no special requirement on the usage amount of the acetic acid, so that the pH value of the second mixed solution can be adjusted. In the present invention, the acetic acid is preferably added in the form of an acetic acid solution, and the mass concentration of the acetic acid is preferably: 90 to 99 percent. The source of the acetic acid is not particularly required in the present invention, and the acetic acid known to those skilled in the art may be used.
In the present invention, the adjustment of the pH is preferably performed under stirring conditions; the stirring speed is preferably 100 to 200rpm, and more preferably 150 to 180 rpm.
In the present invention, the pH adjustment is preferably performed by adding the acetic acid to the second mixed solution at a rate of preferably: 15-20 mL/min. According to the invention, the acetic acid is added into the second mixed solution, and a mixing mode of adding the second mixed solution into the acetic acid is not adopted, so that the levodopa is favorably crystallized as much as possible, and the product yield is favorably improved.
In the invention, the pH value adjustment by using acetic acid preferably comprises a first acidity adjustment and a second acidity adjustment in sequence; the first acidity adjustment further preferably comprises: adjusting the pH value of the second mixed solution to 6-7 by adopting first acetic acid; the second acidity adjustment preferably comprises: and adjusting the pH value of the mixed solution after the first acidity adjustment to 3-5 by adopting second acetic acid.
In the present invention, the first acidity adjustment and the second acidity adjustment are independently preferably performed under stirring conditions; the stirring speed of the first acidity adjustment and the stirring speed of the second acidity adjustment are preferably 100-200 rpm, and more preferably 120-180 rpm independently; the first acidity-adjusted stirring time and the second acidity-adjusted stirring time are independently preferably 1 to 2.5 hours, and more preferably 1.5 to 2 hours.
In the present invention, the first acetic acid and the second acetic acid are independently preferably added in the form of an acetic acid solution; the mass concentrations of the first acetic acid solution and the second acetic acid solution are preferably: 90 to 99 percent.
According to the invention, the pH value of the second mixed solution is adjusted to 6-7 by adopting the first acetic acid, and then the pH value is continuously adjusted to 3-5 by adopting a step-by-step pH value adjusting mode of the second acetic acid, so that the crystal growth of the levodopa is facilitated, the dissolved levodopa can be fully precipitated and crystallized, and the yield of the levodopa is further improved.
After the adjustment of the pH value of the acetic acid is completed, the present invention preferably further includes a crystallization treatment of the mixed solution after the pH value of the acetic acid is adjusted. In the invention, the temperature of the crystallization treatment is preferably 2-7 ℃, and more preferably 3-5 ℃; the time of the crystallization treatment is preferably 2 to 3 hours. In the present invention, the crystallization treatment is preferably: the mixed solution is allowed to stand at the crystallization temperature. The crystallization treatment of the mixed solution can promote the levodopa in the solution to be fully precipitated, and further improve the yield of the levodopa. The method can obtain the levodopa with higher purity and yield by only one-step crystallization, and compared with two-step crystallization, the whole extraction process is simple and feasible, and the product loss is less.
After the levodopa is precipitated, the invention preferably performs solid-liquid separation on the precipitated feed liquid to obtain a solid substance which is the levodopa. The method for separating solid from liquid has no special requirements, and the method for separating solid from liquid can be realized by adopting a solid-liquid separation method which is well known by the technical personnel in the field. The invention preferably adopts a centrifugal separation or filtration separation method to collect solid levodopa.
After the solid-liquid separation, the present invention preferably washes and dries the obtained solid in sequence. In the present invention, the detergent is preferably ethanol and/or acetone. In the present invention, the drying is preferably vacuum drying, and the degree of vacuum of the vacuum drying is preferably-0.07 to-0.1 MPa. In the invention, the drying time is preferably 25-35 min; the drying temperature is preferably 40-50 ℃.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
The method for separating and purifying levodopa provided by the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Preparation of raw material liquid containing levodopa:
firstly, preparing a substrate solution: 10g of catechol, 12g of sodium pyruvate, 45g of ammonium acetate, 2g of sodium sulfite, 1g of ethylenediamine tetraacetic acid, 0.1g of pyridoxal phosphate and 720mL of pure water were mixed, heated to 20 ℃, and dissolved with stirring.
Adding ammonia water into the substrate solution, adjusting the pH value to 8.15, fixing the volume to 900mL, adding 100mL of tyrosine phenol lyase crude enzyme liquid with the mass concentration of 5% sold in the market, sealing the reaction container, filling nitrogen, and carrying out biocatalytic reaction under the conditions of 15 ℃, 200rpm shaking table or stirring.
Except 0min, sampling every hour to measure the residual quantity of catechol and sodium pyruvate, and adding a proper amount of catechol and sodium pyruvate when the content of catechol and sodium pyruvate is low, so that the concentration of catechol is 6g/L and the concentration of sodium pyruvate is 8 g/L.
When the levodopa content did not increase any more, 25ml of 50% sulfuric acid was added to obtain a raw material solution containing levodopa.
500L of raw material liquid containing levodopa is taken, and the concentration of the levodopa in the raw material liquid is 60 g/L. The separation and purification of the raw material liquid comprises the following steps:
1) adding 500g of ascorbic acid into the raw material liquid, uniformly stirring and fully dissolving;
2) slowly adding a sodium hydroxide solution with the mass-volume concentration of 400g/L under the condition that the raw material solution is rapidly stirred at the rotating speed of 400rpm, and adjusting the pH value of the solution to 12 to obtain an alkaline mixed solution; the adding speed of the sodium hydroxide is as follows: 30 mL/min;
3) carrying out low-temperature high-speed centrifugation on the alkaline mixed solution obtained in the step 2) at 4 ℃ and at the rotating speed of 4000rpm for 20min, and collecting the supernatant, wherein the total volume is 500L;
4) adding 15kg of powdered activated carbon into the supernatant obtained in the step 3), heating to 40 ℃, and stirring for 2 hours at the rotating speed of 150 rpm;
5) filtering to remove active carbon, and collecting supernatant;
6) adding 500g of ascorbic acid into the supernatant obtained in the step 5), and fully dissolving to obtain a second mixed solution;
7) dropwise adding the acetic acid solution under the condition of stirring the second mixed solution, wherein the stirring speed is 150rpm, the stirring time is 1 hour, and the pH value of the solution is adjusted to 7.0;
the dropping speed of the acetic acid solution is as follows: 20 mL/min.
8) Continuously dropwise adding an acetic acid solution into the solution with the pH value of 7.0 obtained in the step 7) under the stirring condition, wherein the stirring rotation speed is 150rpm, the stirring time is 1 hour, and the pH value of the solution is adjusted to 4.0;
the dropping speed of the acetic acid solution is as follows: 15 mL/min.
9) Then standing the acidic mixed solution with the pH value of 4.0 obtained in the step 8) at 4 ℃ for 2 hours; carrying out suction filtration to obtain crystals;
10) washing the crystals for 20-30 min by respectively using 300L of ethanol and 300L of acetone in sequence;
11) drying the washed crystal for 30min under vacuum condition at 45 deg.C and-0.07 Mpa to obtain levodopa powder 26.5Kg with yield of 88.3%.
The product obtained in example 1 was subjected to quality testing, and the indexes are shown in table 1 below:
table 1 results of performance test of levodopa prepared in example 1
| Purity of levodopa | 99.8％ |
| Specific rotation degree | -165° |
| Coefficient of absorption | 141 |
| Clarity of the product | Clear and colorless |
| Chloride compound | 0.01％ |
| Loss on drying | 0.4％ |
| Burning residue | 0.06％ |
| Heavy metals | 8ppm |
As can be seen from the data in Table 1, the levodopa separation and purification method provided by the invention has high product yield and purity.
Example 2
A substrate solution is prepared according to the method of the embodiment 1, and the raw material solution with the levodopa content of 70g/L is obtained through the catalysis of the tyrosine phenol lyase. 500L of raw material liquid with the levodopa content of 70g/L is taken, and the separation and purification of the raw material liquid comprises the following steps:
1) adding 500g of ascorbic acid into the raw material liquid, uniformly stirring and fully dissolving;
2) slowly adding a potassium hydroxide solution with the mass-volume concentration of 350g/L under the condition that the raw material solution is rapidly stirred at the rotating speed of 300rpm, and adjusting the pH value of the solution to 12.5 to obtain an alkaline mixed solution; the adding speed of the potassium hydroxide is as follows: 20 mL/min;
3) carrying out low-temperature high-speed centrifugation on the alkaline mixed solution obtained in the step 2) at 5 ℃ and at the rotating speed of 4500rpm for 30min, and collecting the supernatant, wherein the total volume is 500L;
4) adding 20kg of powdered activated carbon into the supernatant obtained in the step 3), heating to 40 ℃, and stirring for 3 hours at the rotating speed of 200 rpm;
5) filtering to remove active carbon, and collecting supernatant;
6) adding 500g of ascorbic acid into the supernatant obtained in the step 5), and fully dissolving to obtain a second mixed solution;
7) dropwise adding the acetic acid solution under the condition of stirring the second mixed solution, wherein the stirring speed is 200rpm, the stirring time is 1.5 hours, and the pH value of the solution is adjusted to 6.5;
the dropping speed of the acetic acid solution is as follows: 20 mL/min;
8) continuously dropwise adding an acetic acid solution into the solution with the pH value of 6.5 obtained in the step 7) under the stirring condition, wherein the stirring speed is 200rpm, the stirring time is 1.5 hours, and the pH value of the solution is adjusted to 3.5;
the dropping speed of the acetic acid solution is as follows: 15 mL/min;
9) then standing the acidic mixed solution with the pH value of 3.5 obtained in the step 8) at 5 ℃ for 3 hours; carrying out suction filtration to obtain crystals;
10) washing the crystal with 350L ethanol and 350L acetone for 30 min;
11) and drying the washed crystal for 30min under vacuum at 50 ℃ and-0.1 Mpa to obtain 31.2Kg of levodopa dry powder with yield of 89.1%.
The product obtained in example 2 was subjected to quality testing, and the indexes are shown in table 2 below:
table 2 results of performance test of levodopa prepared in example 2
| Purity of levodopa | 100％ |
| Specific rotation degree | -162° |
| Coefficient of absorption | 143 |
| Clarity of the product | Clear and colorless |
| Chloride compound | 0.02％ |
| Loss on drying | 0.3％ |
| Burning residue | 0.04％ |
| Heavy metals | 7ppm |
As can be seen from the data in Table 2, the levodopa separation and purification method provided by the invention has high product yield and purity.
Example 3
500L of raw material liquid containing levodopa is taken, and the concentration of the levodopa in the raw material liquid is 50 g/L. The separation and purification of the raw material liquid comprises the following steps:
3) adding 500g of ascorbic acid into the raw material liquid, uniformly stirring and fully dissolving;
4) slowly adding a sodium hydroxide solution with the mass-volume concentration of 380g/L under the condition that the raw material solution is rapidly stirred at the rotating speed of 400rpm, and adjusting the pH value of the solution to 13 to obtain an alkaline mixed solution; the adding speed of the sodium hydroxide is as follows: 25 mL/min;
3) carrying out low-temperature high-speed centrifugation on the alkaline mixed solution obtained in the step 2) at 4 ℃ and at the rotating speed of 4000rpm for 20min, and collecting the supernatant, wherein the total volume is 500L;
4) adding 15kg of powdered activated carbon into the supernatant obtained in the step 3), heating to 40 ℃, and stirring for 2.5 hours at the rotating speed of 150 rpm;
5) filtering to remove active carbon, and collecting supernatant;
6) adding 500g of ascorbic acid into the supernatant obtained in the step 5), and fully dissolving to obtain a second mixed solution;
7) dropwise adding the acetic acid solution under the condition of stirring the second mixed solution, wherein the stirring speed is 150rpm, the stirring time is 1 hour, and the pH value of the solution is adjusted to 6.0;
the dropping speed of the acetic acid solution is as follows: 18 mL/min;
8) continuously dropwise adding an acetic acid solution into the solution with the pH value of 6.0 obtained in the step 7) under the stirring condition, wherein the stirring rotation speed is 150rpm, the stirring time is 1 hour, and the pH value of the solution is adjusted to 3.8;
the dropping speed of the acetic acid solution is as follows: 16 mL/min;
9) then standing the acidic mixed solution with the pH value of 4.0 obtained in the step 8) at 4 ℃ for 2 hours; carrying out suction filtration to obtain crystals;
10) washing the crystals for 20-30 min by respectively using 300L of ethanol and 300L of acetone in sequence;
11) drying the washed crystal for 30min under vacuum condition at 45 deg.C and-0.07 Mpa to obtain 22.7Kg of levodopa powder with yield of 90.8%.
The product obtained in example 3 was subjected to quality testing, and the indexes are shown in table 3 below:
table 3 results of performance test of levodopa prepared in example 3
As can be seen from the data in Table 3, the levodopa separation and purification method provided by the invention has high product yield and purity.
The results of the above examples show that the levodopa separation and purification method provided by the application is simple, purification and separation of levodopa can be realized by only one-step crystallization, and the levodopa separation and purification method has high product yield and purity.
Claims (9)
1. A method for separating and purifying levodopa comprises the following steps:
1) mixing a raw material solution containing levodopa with ascorbic acid to obtain a first mixed solution;
the concentration of levodopa in the raw material liquid is not lower than 30 g/L;
2) adjusting the pH value of the first mixed solution obtained in the step 1) to be alkaline to obtain an alkaline mixed solution;
3) carrying out solid-liquid separation on the alkaline mixed solution obtained in the step 2) to obtain a supernatant;
4) mixing the supernatant obtained in the step 3) with ascorbic acid to obtain a second mixed solution;
5) adjusting the pH value of the second mixed solution obtained in the step 4) to 3-5 by adopting acetic acid, and precipitating levodopa.
2. The separation and purification method according to claim 1, wherein the concentration of the ascorbic acid in the first mixed solution in the step 1) is 0.5 to 2g/L independently from the concentration of the ascorbic acid in the second mixed solution in the step 4).
3. The separation and purification method according to claim 1, wherein the pH value of the alkaline mixed solution in the step 2) is 10 to 13.
4. The separation and purification method according to claim 1, wherein the adjustment of the pH value in the step 5) comprises a first acidity adjustment and a second acidity adjustment in sequence;
the first acidity adjustment comprises: adjusting the pH value of the second mixed solution obtained in the step 4) to 6-7 by adopting first acetic acid;
the second acidity adjustment includes: and adjusting the pH value of the mixed solution after the first acidity adjustment to 3-5 by adopting second acetic acid.
5. The separation and purification method according to claim 1 or 4, further comprising, after the pH adjustment in step 5): and crystallizing the adjusted acidic mixed solution.
6. The separation and purification method according to claim 5, wherein the temperature of the crystallization treatment is 2 to 7 ℃.
7. The separation and purification method according to claim 6, wherein the time for the crystallization treatment is 2 to 3 hours.
8. The separation and purification method according to claim 1, further comprising, before mixing the supernatant obtained in step 3) with ascorbic acid: and (3) decoloring the supernatant by using activated carbon.
9. The separation and purification method according to claim 1, wherein after the levodopa is precipitated in the step 5), solid-liquid separation is performed on the precipitated feed liquid to obtain a solid substance of levodopa.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710711008.2A CN107382760B (en) | 2017-08-18 | 2017-08-18 | Separation and purification method of levodopa |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN103664669A (en) * | 2012-09-05 | 2014-03-26 | 李叶华 | Process method for preparing high-purity levodopa |
| CN106117072A (en) * | 2016-07-08 | 2016-11-16 | 长兴制药股份有限公司 | A kind of levodopa crystalline powder and manufacture method thereof |
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| CN103664669A (en) * | 2012-09-05 | 2014-03-26 | 李叶华 | Process method for preparing high-purity levodopa |
| CN106117072A (en) * | 2016-07-08 | 2016-11-16 | 长兴制药股份有限公司 | A kind of levodopa crystalline powder and manufacture method thereof |
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| 左旋多巴的强降解实验研究;巫世红等;《中国药业》;20081231;第17卷(第21期);20-21 * |
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