Production method of hemin

Abstract

The invention discloses a production method of chlorhematin, which is high in purification rate, high in production utilization rate, high in production safety and free of pollution. The production method is characterized by comprising the following steps: heating reaction, precipitation and crystallization, fractionation and recovery, primary refining and secondary refining. According to the invention, volatilization of acid gas is effectively prevented, and equipment corrosion and environmental pollution are reduced; the reaction temperature in the reaction kettle can be rapidly increased and kept, and the reaction is sufficient; on the premise that the washing effect is guaranteed, the washing and precipitating time is shortened, time is saved, and cleanness and thoroughness are achieved; impurities wrapped in the crystals are effectively removed, and the purification rate is high; the yield of each parallel production line single reaction finished product is increased by dozens of times, and the content of the finished product can reach 97% or above.

Description

Technical Field

[0001] The invention relates to the technical field of biological preparations, in particular to a method for producing hemin. Background Art

[0002] Hemin is an in vitro purified form of natural heme, which is generally separated and purified from animal blood. Heme crystals are widely used in medicine, food, chemical industry, health products, construction and cosmetics industries. In the food industry, hemin can replace nitrite and artificial pigments in meat products. In the cosmetics industry, it is an important raw material. At the same time, heme is also an important raw material for anti-anemia and anti-tumor drugs. In the pharmaceutical industry, it can be used as a semi-synthetic bilirubin raw material, and can be used to prepare anti-cancer special drugs; in clinical practice, it can be made into heme iron supplements. As an iron enhancer, hemin has the advantages of high bioavailability, no adverse reactions such as iron accumulation poisoning and gastrointestinal irritation. This is because hemin is absorbed in molecular form and can be directly taken up by the intestinal mucosa. Hemin is recognized by modern medicine as a biological iron source with high absorption rate and good effect for the prevention and treatment of iron deficiency anemia. It has no iron smell and does not irritate the gastrointestinal tract. It is the first choice for iron and blood supplements for infants and pregnant women.

[0003] Hemoglobin is generally extracted from fresh pig blood, and pig blood cells and plasma are separated by high-speed centrifugation, and then hemoglobin is produced from pig blood cells. Fresh pig blood is not easy to preserve and is easy to deteriorate. When the production volume is small, the plasma produced by separation is also very small, and the transportation utilization value is not high. Generally, it can only be discarded, causing environmental pollution. Then, in order to reduce the loss of raw materials, blood powder, which is relatively easier to preserve, is used. However, since high-concentration volatile acids need to be heated during the production process, this high-temperature and high-concentration volatile acid will corrode the production container equipment and easily cause perforation. For example, the boiling point of glacial acetic acid is 117.9℃, and it is easy to volatilize when heated. When the temperature reaches 108℃, the volatile acid gas not only corrodes the container and the conveying pipeline, but also inevitably destroys the airtightness, escapes and pollutes the air, causing damage to the health of workers and the environment. However, due to the volatilization of the acid due to heat, the temperature rise is limited. Even if the reaction concentration of the acid is increased (to more than 80%), the extraction effect is still not ideal, but the corrosion and loss of the equipment are accelerated. In fact, the content of the hemin obtained in the end can generally only reach 80%, and it is difficult to reach the theoretical content of more than 95% under ideal conditions. The purity of the hemin obtained is difficult to improve because the impurities of the hemin crystals obtained are relatively large. In the preparation process, washing solvents are usually used for purification, and in order to wash more cleanly and thoroughly, a large amount of washing solvents are needed for washing. However, the more washing solvents are used during washing, the longer the time required for precipitation, which seriously affects production efficiency. Summary of the invention

[0004] The purpose of the present invention is to provide a method for producing hemin with high purification rate, high production utilization rate, high production safety and no pollution.

[0005] The present invention adopts the following technical scheme to achieve its invention object, a method for producing hemin, comprising the following steps: ⑴ Heating reaction: Add acetic acid and sodium chloride with a concentration of 70% to 75% into the reaction kettle in sequence according to the ratio, seal it, raise the temperature to 106°C to 108°C, then quickly add pig blood cell powder according to the ratio, keep warm and stir to react for 16 minutes to 18 minutes; Take a sample from the precipitate after the reaction and observe the crystallization under a microscope (xsp-02-640 times). When a rhombus-shaped crystal morphology is formed and blue light can be observed under a darkroom lamp, the reaction is complete. (2) Precipitation and crystallization: After the reaction in step (1) is completed, the reactants are pumped into a precipitation tank for precipitation to allow crystals to precipitate. The solid precipitate obtained is the crude hemin; (3) Fractional distillation recovery: the upper layer liquid after the precipitation separation in step (2) is pumped into a fractionation tank to recover acetic acid; (4) First purification: The crude hemin obtained in step (2) is washed 2 to 3 times with 45% to 50% acetic acid heated to 80°C to 90°C; after cooling, centrifugation is performed to recover the acetic acid liquid, and the separated solid precipitate is washed 2 to 3 times with distilled water and centrifuged to dry; (5) Secondary refining: The dried product obtained in step (4) (i.e., hemin crystals with a content of 93% to 95%) is crushed and passed through a 100-120 mesh sieve; then, it is washed with 30% to 40% acetic acid, distilled water, and 80% alcohol heated to 70° C. to 80° C., and centrifuged and dried 1 to 2 times to promote solubility, further remove impurities in the crude hemin crystals, increase the content, and obtain the hemin finished product.

[0006] The ratio in step (1) of the present invention is based on weight: acetic acid: sodium chloride: pig blood cell powder = 900 kg: 4-5 kg: 75 kg.

[0007] The precipitation time in step (2) of the present invention is 26 to 30 hours; the content of the crude hemin is 70% to 80%.

[0008] The sedimentation tank in step (2) of the present invention comprises a tank body, the bottom of which is inclined (8 to 10 degrees) to facilitate the concentration of sediment; In order to maintain the continuity and efficiency of production, preferably, in the step (2), less than 13 sedimentation tanks are arranged in parallel production lines, and the sedimentation tanks of each parallel production line are connected in series by a pipeline connected to the top of the tank body; The fractionation tank described in step (3) of the present invention is provided with a negative pressure fan, and the negative pressure fan is connected to a collection tower via a pipeline; the acetic acid volatile gas in the fractionation tank is transported to the collection tower through the negative pressure fan through the side pipe pipeline, and a sodium hydroxide solution is sprayed in the collection tower to carry out a neutralization reaction to obtain a sodium acetate solution for preparing industrial-grade sodium acetate.

[0009] In step (4) of the present invention, the crude hemin is washed with acetic acid to obtain hemin crystals with a content of 85% to 90%, and then washed with distilled water to obtain hemin crystals with a content of 93% to 95%; The amount of distilled water used for washing in step (4) is 5 to 8 times the weight of the solid precipitate separated by centrifugation.

[0010] The water content of the hemin finished product in step (5) of the present invention is ≤0.4%; the water content of the hemin finished product obtained in step (5) is 98% to 99%.

[0011] In step (3) of the present invention, the precipitate after fractionation in the fractionation tank is adjusted to a pH value of 6.8 to 7.0 (using sodium hydroxide solution), and then dried to obtain globulin as feed; The sodium hydroxide solution is a sodium hydroxide solution with a pH value of 7 to 9 and a concentration of 3% to 4%.

[0012] Due to the adoption of the above technical scheme, the present invention achieves the purpose of the invention better, which effectively prevents the volatilization of acidic gases, reduces equipment corrosion and environmental pollution; can quickly raise the reaction temperature in the reactor to maintain heat preservation, and fully react; under the premise of ensuring the washing effect, shortens the washing and precipitation time, saves time, and is clean and thorough; effectively removes impurities wrapped in crystals, and has a high purification rate; the amount of finished products obtained in a single reaction of each parallel production line increases by dozens of times, and the content of the finished product can reach more than 97%. DETAILED DESCRIPTION

[0013] The present invention will be further described below in conjunction with the embodiments.

[0014] Embodiment 1: A method for producing hemin comprises the following steps: (1) Heating reaction: add acetic acid with a concentration of 70% to 75% (70% acetic acid in this embodiment) and sodium chloride into the reaction kettle in sequence according to the ratio, seal it, raise the temperature to 106°C to 108°C (106°C in this embodiment), then quickly add pig blood cell powder according to the ratio, keep warm and stir to react for 16 minutes to 18 minutes (16 minutes in this embodiment); Take a sample from the precipitate after the reaction and observe the crystallization under a microscope. When a rhombus-shaped crystal morphology is formed and blue light can be observed under a darkroom lamp, the reaction is complete. (2) Precipitation and crystallization: After the reaction in step (1) is completed, the reactants are pumped into a precipitation tank for precipitation to allow crystals to precipitate. The solid precipitate obtained is the crude hemin; (3) Fractional distillation recovery: the upper layer liquid after the precipitation separation in step (2) is pumped into a fractionation tank to recover acetic acid; (4) First purification: The crude hemin obtained in step (2) is heated to 80°C to 90°C (80°C in this embodiment) with 45% to 50% acetic acid (45% acetic acid in this embodiment) and washed 2 to 3 times (3 times in this embodiment); after cooling, centrifugation is performed to recover the liquid, and the separated solid precipitate is washed 2 to 3 times (2 times in this embodiment) with distilled water, and centrifuged and dried; (5) Secondary refining: The dried product obtained in step (4) is crushed and sieved; then, 30% to 40% acetic acid (30% acetic acid in this embodiment), distilled water, and 80% alcohol heated to 70° C. to 80° C. (70° C. in this embodiment) are used for washing and centrifugal drying for 1 to 2 times (once in this embodiment) to promote solubility, further remove impurities in the crystals of the crude hemin, increase the content, and obtain the finished hemin.

[0015] The ratio in step (1) of the present invention is based on weight, acetic acid: sodium chloride: pig blood cell powder = 900kg: 4-5kg: 75kg (900kg: 4kg: 75kg in this embodiment).

[0016] The precipitation time in step (2) of the present invention is 26 to 30 hours (26 hours in this embodiment); the content of the crude hemin is 70% to 80% (70% in this embodiment).

[0017] The sedimentation tank in step (2) of the present invention comprises a tank body, and the bottom of the tank body is inclined; The fractionation tank described in step (3) of the present invention is provided with a negative pressure fan, and the negative pressure fan is connected to a collection tower via a pipeline; the acetic acid volatile gas in the fractionation tank is transported to the collection tower through the negative pressure fan through the side pipe pipeline, and a sodium hydroxide solution is sprayed in the collection tower to carry out a neutralization reaction to obtain a sodium acetate solution for preparing sodium acetate.

[0018] In step (4) of the present invention, the crude hemin is washed with acetic acid to obtain a content of 85% to 90% (85% in this embodiment), and then washed with distilled water to obtain a content of 93% to 95% (93% in this embodiment); The distilled water used for washing in step (4) is 5 to 8 times (5 times in this embodiment) the weight of the solid precipitate separated by centrifugation.

[0019] The water content of the hemin finished product in step ⑸ of the present invention is ≤0.4% (0.4% in this embodiment); the water content of the hemin finished product obtained in step ⑸ is 98% to 99% (98% in this embodiment).

[0020] In step (3) of the present invention, the pH value of the precipitate after distillation in the fractionation tank is adjusted to 6.8-7.0 (6.8 in this embodiment), and then dried to obtain globulin as feed. Embodiment 2:

[0021] In step (1) of this embodiment, acetic acid with a concentration of 70% to 75% (75% acetic acid in this embodiment) and sodium chloride are added to the reaction kettle in sequence according to the ratio, sealed, and heated to 106° C. to 108° C. (108° C. in this embodiment), and then pig blood cell powder is quickly added according to the ratio, and the reaction is stirred and kept warm for 16 minutes to 18 minutes (18 minutes in this embodiment); In step (4), the crude hemin obtained in step (2) is heated to 80° C. to 90° C. (90° C. in this embodiment) with 45% to 50% acetic acid (50% acetic acid in this embodiment) and washed 2 to 3 times (twice in this embodiment); after cooling, centrifugation is performed to recover the liquid, and the separated solid precipitate is washed 2 to 3 times (three times in this embodiment) with distilled water, and centrifuged and dried; In step (5), the dried product obtained in step (4) is crushed and sieved; then, 30% to 40% acetic acid (40% acetic acid in this embodiment), distilled water, and 80% alcohol heated to 70° C. to 80° C. (80° C. in this embodiment) are used for washing and centrifugal drying for 1 to 2 times (2 times in this embodiment) to obtain a finished hemin.

[0022] In this embodiment, the ratio in step (1) is based on weight, acetic acid: sodium chloride: pig blood cell powder = 900kg: 4-5kg: 75kg (900kg: 5kg: 75kg in this embodiment).

[0023] In this embodiment, the precipitation time in step (2) is 26 hours to 30 hours (30 hours in this embodiment); the content of the crude hemin is 70% to 80% (80% in this embodiment).

[0024] In this embodiment, the crude hemin in step (4) is washed with acetic acid to obtain a content of 85% to 90% (88% in this embodiment), and then washed with distilled water to obtain a content of 93% to 95% (94.5% in this embodiment); The distilled water used for washing in step (4) is 5 to 8 times (6 times in this embodiment) the weight of the solid precipitate separated by centrifugation.

[0025] In this embodiment, the water content of the hemin finished product in step ⑸ is ≤0.4% (0.2% in this embodiment); the content of the hemin finished product obtained in step ⑸ is 98% to 99% (98% in this embodiment).

[0026] In this embodiment, in the step (3), the pH value of the precipitate after distillation in the distillation tank is adjusted to 6.8-7.0 (7.0 in this embodiment), and then dried to obtain globulin as feed.

[0027] The rest is the same as in Example 1. Embodiment 3:

[0028] In step (1) of this embodiment, acetic acid with a concentration of 70% to 75% (73% acetic acid in this embodiment) and sodium chloride are added to the reaction kettle in sequence according to the ratio, sealed, and heated to 106°C to 108°C (107°C in this embodiment), and then pig blood cell powder is quickly added according to the ratio, and the reaction is stirred and kept warm for 16 minutes to 18 minutes (17 minutes in this embodiment); In step (4), the crude hemin obtained in step (2) is heated to 80° C. to 90° C. (85° C. in this embodiment) with 45% to 50% acetic acid (47% acetic acid in this embodiment) and washed 2 to 3 times (3 times in this embodiment); after cooling, centrifugation is performed to recover the liquid, and the separated solid precipitate is washed 2 to 3 times (3 times in this embodiment) with distilled water, and centrifuged and dried; In step (5), the dried product obtained in step (4) is crushed and sieved; then, 30% to 40% acetic acid (35% acetic acid in this embodiment), distilled water, and 80% alcohol heated to 70° C. to 80° C. (75° C. in this embodiment) are used for washing and centrifugal drying for 1 to 2 times (twice in this embodiment) to obtain a finished hemin.

[0029] In this embodiment, the ratio in step (1) is based on weight, acetic acid: sodium chloride: pig blood cell powder = 900kg: 4-5kg: 75kg (900kg: 4.5kg: 75kg in this embodiment).

[0030] In this embodiment, the precipitation time in step (2) is 26 to 30 hours (28 hours in this embodiment); the content of the crude hemin is 70% to 80% (75% in this embodiment).

[0031] In this embodiment, the crude hemin in step (4) is washed with acetic acid to obtain a content of 85% to 90% (90% in this embodiment), and then washed with distilled water to obtain a content of 93% to 95% (95% in this embodiment); The distilled water used for washing in step (4) is 5 to 8 times (8 times in this embodiment) the weight of the solid precipitate separated by centrifugation.

[0032] In this embodiment, the water content of the hemin finished product in step ⑸ is ≤0.4% (0.16% in this embodiment); the content of the hemin finished product obtained in step ⑸ is 98% to 99% (98.6% in this embodiment).

[0033] In this embodiment, in the step (3), the pH value of the precipitate after distillation in the distillation tank is adjusted to 6.8-7.0 (6.9 in this embodiment), and then dried to obtain globulin as feed.

[0034] The rest is the same as in Example 1. Embodiment 4:

[0035] In this embodiment, step (1) adds acetic acid with a concentration of 70% to 75% (73% acetic acid in this embodiment) and sodium chloride into the reaction kettle in sequence according to the ratio, seals it, raises the temperature to 106° C. to 108° C. (107° C. in this embodiment), then quickly adds pig blood cell powder according to the ratio, and keeps the temperature and stirs to react for 16 minutes to 18 minutes (17 minutes in this embodiment); In step (4), the crude hemin obtained in step (2) is heated to 80° C. to 90° C. (85° C. in this embodiment) with 45% to 50% acetic acid (48% acetic acid in this embodiment) and washed 2 to 3 times (twice in this embodiment); after cooling, centrifugation is performed to recover the liquid, and the separated solid precipitate is washed 2 to 3 times (three times in this embodiment) with distilled water and centrifuged to dry; In step (5), the dried product obtained in step (4) is crushed and sieved; then, 30% to 40% acetic acid (40% acetic acid in this embodiment), distilled water, and 80% alcohol heated to 70° C. to 80° C. (75° C. in this embodiment) are used for washing and centrifugal drying for 1 to 2 times (2 times in this embodiment) to obtain a finished hemin.

[0036] In this embodiment, the ratio in step (1) is based on weight, acetic acid: sodium chloride: pig blood cell powder = 900kg: 4-5kg: 75kg (900kg: 4.5kg: 75kg in this embodiment).

[0037] In this embodiment, the precipitation time in step (2) is 26 to 30 hours (28 hours in this embodiment); the content of the crude hemin is 70% to 80% (75% in this embodiment).

[0038] In this embodiment, the crude hemin in step (4) is washed with acetic acid to obtain a content of 85% to 90% (90% in this embodiment), and then washed with distilled water to obtain a content of 93% to 95% (95% in this embodiment); The distilled water used for washing in step (4) is 5 to 8 times (8 times in this embodiment) the weight of the solid precipitate separated by centrifugation.

[0039] In this embodiment, the water content of the hemin finished product in step ⑸ is ≤0.4% (0.12% in this embodiment); the content of the hemin finished product obtained in step ⑸ is 98% to 99% (99% in this embodiment).

[0040] In this embodiment, in the step (3), the pH value of the precipitate after distillation in the distillation tank is adjusted to 6.8-7.0 (6.9 in this embodiment), and then dried to obtain globulin as feed.

[0041] The rest is the same as in Example 1.

[0042] Test example 1. Experimental treatment methods and grouping 1. Control group (A): S1. Take 100kg of fresh pig blood and 0.5kg of sodium citrate, mix and stir, cool for 10 minutes, centrifuge at 5000rpm to separate fresh pig blood cells, and store at 3°C ​​for use; S2. Take 1200L of 75% acetic acid and heat it to 90°C, add 9kg of sodium chloride and completely dissolve it to obtain an acetate solution for use; S3. Add acetate solution to fresh pig blood cell solution at a ratio of fresh pig blood cell solution to acetate solution = 0.4:1, and stir and react at 90°C for 50 minutes; then S4. Stirring and vacuum filtering to obtain a solid solid for solid-liquid separation as crude hemin; S5. 200 L of 60% acetic acid was added to the crude hemin for washing, and then pumped into a vacuum filter and stirred for 45 minutes, and then vacuum filtered for solid-liquid separation; the obtained solid was washed three times with 100 L of deionized water, and each washing was stirred for 10 minutes, and then vacuum filtered for solid-liquid separation; The obtained solid was added with 60% acetic acid and this step was repeated 3 times; S6. Finally, the hemin obtained by vacuum filtration is dried to obtain the hemin finished product.

[0043] ⒉ (B) to (E) are respectively embodiments 1 to 4 of the present invention.

[0044] 2. Comparison of test results The purity of the hemin products obtained by (A) to (E) was tested, and the test results are as follows: Table 1. Purity test of hemin

[0045] 3. Product performance testing and evaluation Samples (100g / bag) were taken from component E and tested according to the evaluation criteria of "National Food Safety Standard Food Nutrition Fortifier Hemin Chloride". The test results are as follows: Table 2. Evaluation of hemin

[0046] The above experiments prove that the present invention can improve the product purity of hemin well, and the evaluation results are qualified and the product quality is high.

Claims

1. A method for producing hemin, Features The following steps are involved: ⑴ Heating reaction: Add acetic acid and sodium chloride with a concentration of 70% to 75% into the reaction kettle in sequence according to the ratio, seal it, raise the temperature to 106°C to 108°C, then quickly add pig blood cell powder according to the ratio, keep warm and stir to react for 16 minutes to 18 minutes; Take a sample from the precipitate after the reaction and observe the crystallization under a microscope. When a rhombus-shaped crystal morphology is formed and blue light can be observed under a darkroom lamp, the reaction is complete. (2) Precipitation and crystallization: After the reaction in step (1) is completed, the reactants are pumped into a precipitation tank for precipitation to allow crystals to precipitate. The solid precipitate obtained is the crude hemin; (3) Fractional distillation recovery: the upper layer liquid after the precipitation separation in step (2) is pumped into a fractionation tank to recover acetic acid; (4) First purification: The crude hemin obtained in step (2) is washed 2 to 3 times with 45% to 50% acetic acid heated to 80°C to 90°C; after cooling, centrifugation is performed to recover the liquid, and the separated solid precipitate is washed 2 to 3 times with distilled water and centrifuged to dry; (5) Secondary refining: The dried product obtained in step (4) is crushed and sieved; then, it is washed with 30% to 40% acetic acid, distilled water, and 80% alcohol heated to 70°C to 80°C, respectively, and centrifuged and dried 1 to 2 times to obtain the finished hemin.

2. A method for producing hemin according to claim 1, Features The ratio in step (1) is by weight: acetic acid: sodium chloride: pig blood cell powder = 900 kg: 4-5 kg: 75 kg.

3. A method for producing hemin according to claim 1, Features The precipitation time in step (2) is 26 to 30 hours; the content of the crude hemin is 70% to 80%.

4. A method for producing hemin according to claim 2, Features The precipitation time in step (2) is 26 to 30 hours; the content of the crude hemin is 70% to 80%.

5. A method for producing hemin according to claim 3, Features The precipitation tank in step (2) comprises a tank body, the bottom of which is inclined; The fractionation tank described in step (3) is provided with a negative pressure fan, and the negative pressure fan is connected to a collection tower via a pipeline; the acetic acid volatile gas in the fractionation tank is transported to the collection tower through the negative pressure fan through the side pipe pipeline to the collection tower, and a sodium hydroxide solution is sprayed in the collection tower to carry out a neutralization reaction to obtain a sodium acetate solution for preparing sodium acetate.

6. A method for producing hemin according to claim 4, Features The precipitation tank in step (2) comprises a tank body, the bottom of which is inclined; The fractionation tank described in step (3) is provided with a negative pressure fan, and the negative pressure fan is connected to a collection tower via a pipeline; the acetic acid volatile gas in the fractionation tank is transported to the collection tower through the negative pressure fan through the side pipe pipeline to the collection tower, and a sodium hydroxide solution is sprayed in the collection tower to carry out a neutralization reaction to obtain a sodium acetate solution for preparing sodium acetate.

7. A method for producing hemin according to claim 1 or 2 or 3 or 4 or 5 or 6, Features In the step (4), the crude hemin is washed with acetic acid to obtain a content of 85% to 90%, and then washed with distilled water to obtain a content of 93% to 95%; The amount of distilled water used for washing in step (4) is 5 to 8 times the weight of the solid precipitate separated by centrifugation.

8. A method for producing hemin according to claim 1 or 2 or 3 or 4 or 5 or 6, Features The water content of the hemin finished product in step (5) is ≤0.4%; the water content of the hemin finished product obtained in step (5) is 98% to 99%.

9. A method for producing hemin according to claim 7, Features The water content of the hemin finished product in step (5) is ≤0.4%; the water content of the hemin finished product obtained in step (5) is 98% to 99%.

10. A method for producing hemin according to claim 9, Features In the step (3), the pH value of the precipitate after distillation in the distillation tank is adjusted to 6.8-7.0, and then dried to obtain globulin as feed.

Images