CN107056884B - Analytic process for purifying glutathione metal salt - Google Patents
Analytic process for purifying glutathione metal salt Download PDFInfo
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Abstract
The invention belongs to the technical field of medicines, and relates to an analytic process for purifying reduced glutathione metal salt. The process adopts a sodium sulfide mixture containing 10-50% of sodium chloride as an analytical agent to replace hydrogen sulfide gas, and the sodium sulfide mixture reacts with glutathione metal salt to analyze the glutathione. Compared with the process of adding hydrogen sulfide, the process has the following advantages: the safety is improved, and huge potential safety hazards in the transportation, storage and use processes of the hydrogen sulfide are avoided; the resolving agent in the process can effectively improve the reaction yield; compared with hydrogen sulfide, the sodium sulfide and sodium chloride mixed solid is cheaper in price and the production cost is reduced.
Description
Technical Field
The invention belongs to the technical field of medicines, and relates to an analytic process of glutathione metal salt.
Background
Glutathione (GSH) is a tripeptide containing gamma-amide bonds and sulfhydryl groups, and is composed of glutamic acid, cysteine and glycine. Present in almost every cell of the body. Glutathione can help to maintain normal immune system function, and has antioxidant and antidotal effects.
At present, glutathione is produced by fermentation in the prior art, and after production is finished, yeast cells are broken and then glutathione is obtained by extraction. The existing glutathione purification technology is that glutathione in a multi-impurity extracting solution is converted into glutathione metal salt by using metal salt, the glutathione metal salt is separated and washed, the glutathione metal salt is dispersed in water, hydrogen sulfide with stronger bonding force with metal ions is added to generate water-soluble glutathione and water-insoluble metal sulfide precipitate, and the glutathione aqueous solution is obtained after filtration.
Taking glutathione cuprous salt as an example, the analytical reaction equation is as follows:
2CuGS+H2S→Cu2S↓+2GSH
the prior art analysis agent is hydrogen sulfide gas, and patents JP52-73821 and JP62-283994 describe the analysis of reduced glutathione by hydrogen sulfide gas. The hydrogen sulfide is a 2.1-class flammable gas, and safety accidents easily occur in the production process; compared with solid materials, the material is particularly easy to leak; meanwhile, the gas is 2.3-class toxic gas, and has strong toxicity and occasionally poisoning. Particularly, in the process of large-scale production, hydrogen sulfide gas has huge potential safety hazards in the transportation, storage and use processes.
Hydrogen sulfide is a binary weak acid. At 20 ℃, 1 volume of water can dissolve 2.6 volumes of hydrogen sulfide, and the resulting aqueous solution is called hydrosulfuric acid and has a concentration of 0.1 mol/L. Therefore, the reaction of hydrogen sulfide with the complex is limited, the final yield of the reaction is about 85% of the theoretical value, about 10% of reduced glutathione is not resolved, and a certain amount of waste is caused.
Meanwhile, hydrogen sulfide is used as a resolving agent, and the price is higher in the using process, so that the large production cost is higher; because the yield of the glutathione is only 85 percent when the hydrogen sulfide is used for the analytic reaction, the production cost is further promoted, and the product cost is greatly increased in the mass production with more key production cost.
The sodium sulfide is solid, and can be used as an analytical agent, so that the defects of hydrogen sulfide gas, such as difficult storage, high toxicity and the like, can be solved. According to the mechanism of chemical reaction, the problem of poor safety, low yield and high cost of the hydrogen sulfide can be solved by considering that sodium sulfide is directly used for replacing hydrogen sulfide to solve the defects in the production process of the hydrogen sulfide, however, the sodium sulfide is a strong base substance and is added into a complex to greatly increase the pH value of the solution, if the reaction speed is not fast enough, the generated product reduced glutathione can be catalyzed by dissolved oxygen under the catalysis of unreacted metal ions such as copper ions to generate oxidized glutathione, so that the yield and the purity are reduced, so that the analytic method for replacing the hydrogen sulfide by the sodium sulfide has not been reported and patented so far, and the invention needs to invent a solid reagent to accelerate the reaction speed, so that the analyzed metal ions can be quickly complexed by the sulfur ions in a very short time and cannot play a role in catalysis.
Disclosure of Invention
The inventor adopts mixed solid of sodium sulfide and sodium chloride to replace hydrogen sulfide for analysis through research, development and innovation, wherein the weight percentage of sodium chloride in the analysis agent is 10% -50%, and the optimal weight percentage of sodium chloride in the analysis agent is 15%. Sodium sulfide contains sulfide ions and is a good resolving agent, but sodium sulfide is a strong alkaline substance, and the resolved product reduced glutathione is a strong reducing substance, so that oxidized glutathione is easily generated by using sodium sulfide alone, thereby reducing the yield.
After sodium chloride is added into sodium sulfide, the unexpected discovery can be through the complex of the dissolved effect of neutral strong electrolyte sodium chloride, increase reaction rate to reduce the proportion of oxidized glutathione, improved product purity and improved yield. After the desorption agent is replaced by the mixed solid of sodium sulfide and sodium chloride, the solid has small toxic and side effects, so that high cost and high risk in the transportation and storage processes are avoided, and the yield of the desorption reaction is obviously higher than that of the existing process using hydrogen sulfide as the desorption agent.
The analytic process flow chart of the invention is as follows:
the process is described below, using glutathione cuprous as an example, but it applies to all glutathione metal salts:
2CuGS+Na2S+2H+→2GSH+Gu2S↓+2Na+
and filtering and centrifuging to remove copper sulfide to obtain glutathione resolution solution.
The analytic process of the invention adopts sodium sulfide solid added with sodium chloride as an analytic agent, wherein the glutathione metal salt is not limited to any metal, is especially suitable for the heavy metal salt in pharmacy, such as silver, lead, iron, mercury and the like, and can be used for analyzing glutathione according to the reaction principle of the glutathione metal salt and cuprous glutathione, thereby obtaining the reduced glutathione.
The proportion of sodium chloride in the resolving agent is important, generally 10% -50%, glutathione can be resolved, but the yield of the resolving agent is affected by the proportion of sodium chloride, and the results obtained by a plurality of tests are shown in the following table:
influence of different ratios of sodium chloride on the analytical yield:
| sodium chloride ratio in the resolving agent | Analytical yield |
| 5％ | 91.7％ |
| 10％ | 95.0％ |
| 15％ | 97.2％ |
| 50％ | 95.5％ |
| 60％ | 90.2％ |
The application of the mixture of sodium sulfide and sodium chloride in the preparation of the glutathione metal salt resolving agent is used as a new resolving agent, plays a role in the resolving process of the glutathione metal salt, and has the characteristics of high safety, high yield, low cost and the like, and comprises the following steps:
(1) the mixed solid of sodium sulfide and sodium chloride is added into glutathione metal salt for reaction, so that the safety is improved. Compared with hydrogen sulfide gas, the mixed solid of sodium sulfide and sodium chloride can not generate highly toxic hydrogen sulfide gas in the storage process, can not be ignited, occupies less storage space, replaces flammable, explosive and highly toxic hydrogen sulfide gas, and improves the overall safety in the aspects of transportation, storage, use and the like.
(2) The mixed solid of sodium sulfide and sodium chloride is added into glutathione metal salt for reaction, so that the reaction yield is improved. Hydrogen sulfide is a binary weak acid. 1 volume of water at 20 ℃ can dissolve 2.6 volumes of hydrogen sulfide, and the resulting aqueous solution is called hydrosulfuric acid and has a concentration of 0.1 mol/L. Therefore, the reaction of hydrogen sulfide with the complex is limited, and the final yield of the reaction is about 85% of the theoretical value. The mixed solid of sodium sulfide and sodium chloride is easy to dissolve in water, and the solubility is more than 4mol/L and far more than hydrogen sulfide under the same condition. The final reaction yield of the mixed solid of sodium sulfide and sodium chloride can reach about 95-97%, which is improved by about 10% compared with the prior art, the solution is almost complete, and the cost is greatly reduced in industrial production.
(3) The mixed solid of sodium sulfide and sodium chloride is added into glutathione metal salt for reaction, and compared with hydrogen sulfide, the price is cheaper, and the production cost is low.
The cost of the resolving agent is compared with that of hydrogen sulfide and sodium sulfide resolving agents:
through a plurality of experiments of the inventor, the mixture of sodium sulfide and sodium chloride can be applied to the analysis agent of the reduced glutathione metal salt. The metal salt is preferably a pharmaceutically-mentioned heavy metal salt, which has lower activity than hydrogen ions and can be replaced by the hydrogen ions, and the copper ions are most commonly used in the pharmaceutical industry. Meanwhile, in the mixture of the sodium sulfide and the sodium chloride, the weight percentage of the sodium chloride is 10-50%, and the preferred weight percentage is 15%.
Detailed Description
The present invention will now be described in detail by way of specific examples, which are to be construed as merely illustrative, and not a limitation of the present invention.
Example 1: and (3) when hydrogen sulfide is used as a resolving agent, resolving reduced glutathione cuprous:
the cuprous glutathione is dispersed in the purified water, the total volume is 1100L, the glutathione content is 1.33 percent, and the total amount of the glutathione is 14.63 kg. Hydrogen sulfide gas is introduced into the solution at the temperature of between 15 and 25 ℃ under stirring, the flow rate is about 2kg/h, and the color of the solution is changed from milky white to black. After 30 minutes, the glutathione content in the analytical solution was 1.15% by sampling. Stopping introducing hydrogen sulfide, sealing and stirring for 1 hour, adjusting the pH of the analysis solution to 4.0 +/-0.2 by using 10% sodium hydroxide solution or hydrochloric acid, sampling and detecting that the content of glutathione in the analysis solution is 1.17%, the purity is 93.7% and the yield is 85.3%. The weight reduction method measures that the hydrogen sulfide is actually used in a hydrogen sulfide steel cylinder by 0.97kg, and the cost of the analysis agent is 97.00.
Example 2: when sodium sulfide solid is used as an analysis agent, analyzing the cuprous glutathione:
the cuprous glutathione is dispersed in the purified water, the total volume is 1100L, the glutathione content is 1.38 percent, and the total amount of the glutathione is 15.18 kg. 2.03kg of sodium sulfide solid is added under stirring at 15-25 ℃. After 10 minutes, the solution turns black, after stirring for 30 minutes, the pH value of the analysis solution is adjusted to 4.0 +/-0.2 by using 10% sodium hydroxide solution or hydrochloric acid, the content of glutathione in the analysis solution is 1.23% through sampling detection, the purity is 85.2%, and the yield is 84.1%. 2.03kg of sodium sulfide was added in total, the cost of the desorption agent was 30.25, and the purity was lower than that of hydrogen sulfide as the desorption agent.
Example 3: when a mixed solid of sodium sulfide and sodium chloride (10%) is used as an analytical agent, the cuprous glutathione is analyzed:
the cuprous glutathione is dispersed in the purified water, the total volume is 1100L, the glutathione content is 1.40 percent, and the total amount of the glutathione is 15.40 kg. 2.29kg of mixed solid of sodium sulfide and sodium chloride is added under stirring at 15-25 ℃, and the sodium chloride accounts for 10 percent. After 5 minutes, the solution turns black, after stirring for 30 minutes, the pH value of the analysis solution is adjusted to 4.0 +/-0.2 by using 10% sodium hydroxide solution or hydrochloric acid, the content of glutathione in the analysis solution is 1.33% through sampling detection, the purity is 92.4%, and the yield is 95.0%. The amount of sodium sulfide charged was 2.061kg, the amount of sodium chloride was 0.229kg, and the cost of the analytical agent was about 30.78.
Example 4: when a mixed solid of sodium sulfide and sodium chloride (15%) is used as an analytical agent, glutathione cuprous is analyzed:
the cuprous glutathione is dispersed in the purified water, the total volume is 1100L, the glutathione content is 1.44%, and the total amount of the glutathione is 15.84 kg. 2.49kg of mixed solid of sodium sulfide and sodium chloride is added under stirring at 15-25 ℃, wherein the sodium chloride accounts for 15 percent. After 5 minutes, the solution turns black, after stirring for 30 minutes, the pH value of the analysis solution is adjusted to 4.0 +/-0.2 by using 10% sodium hydroxide solution or hydrochloric acid, the content of glutathione in the analysis solution is 1.40% through sampling detection, the purity is 94.5%, and the yield is 97.2%. The amount of sodium sulfide charged was 2.12kg, the amount of sodium chloride charged was 0.37kg, and the cost of the analyzing agent was about 31.72.
Example 5: when a mixed solid of sodium sulfide and sodium chloride (50%) is used as an analytical agent, glutathione cuprous is analyzed:
the cuprous glutathione is dispersed in the purified water, the total volume is 1100L, the glutathione content is 1.41 percent, and the total amount of the glutathione is 15.51 kg. 4.14kg of mixed solid of sodium sulfide and sodium chloride is added under stirring at 15-25 ℃, wherein the sodium chloride accounts for 50 percent. After 5 minutes, the solution turns black, after stirring for 30 minutes, the pH value of the analysis solution is adjusted to 4.0 +/-0.2 by using 10% sodium hydroxide solution or hydrochloric acid, the content of glutathione in the analysis solution is 1.35% through sampling detection, the purity is 94.1%, and the yield is 95.5%. The amount of sodium sulfide charged was 2.07kg, the amount of sodium chloride charged was 2.07kg, and the cost of the analyzing agent was about 32.46.
The quality and yield of the copper salt analytic products produced by various methods are shown in the table:
therefore, the yield of the hydrogen sulfide used as the resolving agent is low, and the resolving cost is high; however, sodium sulfide is not used as the resolving agent in the current industrial mass production because a large amount of oxidized glutathione is generated in the resolving process, the purity of the sample is poor, and the subsequent refining is troublesome. The mixture of sodium sulfide and sodium chloride can solve the problem of the deficiency of sodium monosulfide, the resolution cost is lower, the resolution yield is higher than that of hydrogen sulfide, and unexpected technical effects are generated.
Example 6: examples of other metallic salts of glutathione
2AgGS+Na2S+2H+→2GSH+Ag2S↓+2Na+
When a mixed solid of sodium sulfide and sodium chloride (15%) is used as an analytical agent, glutathione silver salt is analyzed:
the prepared glutathione silver salt complex is dispersed in purified water, the total volume is 1L, the glutathione content is 1.36 percent, and the total amount of glutathione is 13.6 g. 4.07g of a mixed solid of sodium sulfide and sodium chloride was added at room temperature, the sodium chloride accounting for 15%. After 5 minutes, the color of the solution turns black, the solution is continuously stirred for 30 minutes, and then the content of glutathione in the analysis solution is sampled and detected to be 1.29 percent, and the yield is 94.9 percent. The amount of sodium sulfide charged was 3.46g, and the amount of sodium chloride charged was 0.61 kg.
Claims (7)
1. A process for resolving reduced glutathione metal salt, which is characterized by comprising the following steps: the resolving agent used in the resolving process consists of sodium sulfide and sodium chloride.
2. The analytical process according to claim 1, wherein: the reduced glutathione metal salt is the heavy metal salt in pharmacy.
3. The analytical process according to claim 2, wherein: the reduced glutathione metal salt is reduced glutathione cuprous salt.
4. Analytical process according to any one of claims 1 to 3, characterised in that: the weight percentage of sodium chloride in the resolving agent is 10-50%.
5. The analytical process according to claim 4, wherein: the weight percentage of sodium chloride in the resolving agent was 15%.
6. The application of the mixture of sodium sulfide and sodium chloride in preparing the analytical agent of reduced glutathione metal salt.
7. The use of claim 6, wherein the reduced glutathione metal salt is a cuprous salt.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429229A (en) * | 2008-12-14 | 2009-05-13 | 甘肃正生生物科技有限公司 | Method for producing high-purity glutathione |
| CN106008664A (en) * | 2016-07-26 | 2016-10-12 | 上海青平药业有限公司 | Efficient separation and purification method of glutathione |
| CN106220708A (en) * | 2016-07-26 | 2016-12-14 | 上海青平药业有限公司 | A kind of method extracting separating glutathione from GSCu precipitates |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429229A (en) * | 2008-12-14 | 2009-05-13 | 甘肃正生生物科技有限公司 | Method for producing high-purity glutathione |
| CN106008664A (en) * | 2016-07-26 | 2016-10-12 | 上海青平药业有限公司 | Efficient separation and purification method of glutathione |
| CN106220708A (en) * | 2016-07-26 | 2016-12-14 | 上海青平药业有限公司 | A kind of method extracting separating glutathione from GSCu precipitates |
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