CN112999897A - Method and process for extracting heme from blood meal by using reverse osmosis membrane - Google Patents
Method and process for extracting heme from blood meal by using reverse osmosis membrane Download PDFInfo
- Publication number
- CN112999897A CN112999897A CN202110299597.4A CN202110299597A CN112999897A CN 112999897 A CN112999897 A CN 112999897A CN 202110299597 A CN202110299597 A CN 202110299597A CN 112999897 A CN112999897 A CN 112999897A
- Authority
- CN
- China
- Prior art keywords
- membrane
- reverse osmosis
- acetone
- heme
- osmosis membrane
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
- B01D71/06—Organic material
- B01D71/08—Polysaccharides
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/02—Reverse osmosis; Hyperfiltration ; Nanofiltration
- B01D61/025—Reverse osmosis; Hyperfiltration
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
- B01D67/0002—Organic membrane manufacture
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
- B01D69/12—Composite membranes; Ultra-thin membranes
- B01D69/125—In situ manufacturing by polymerisation, polycondensation, cross-linking or chemical reaction
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
- B01D71/06—Organic material
- B01D71/56—Polyamides, e.g. polyester-amides
Landscapes
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Water Supply & Treatment (AREA)
- Nanotechnology (AREA)
- Manufacturing & Machinery (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
Abstract
The invention discloses a method for extracting heme from blood powder by using reverse osmosis technology, which comprises the steps of using animal blood powder as a raw material, particularly pig blood powder, dissolving the raw material by using purified water, adding a protein precipitator into a mixed solution, collecting precipitates after filtering, adding acidic acetone into the precipitates, performing suction filtration after dissolving, collecting filtrate, filtering the filtrate by using a reverse osmosis membrane to obtain concentrated heme, and drying to obtain high-purity heme. The method is characterized in that a protein precipitator is used for accelerating the separation of protein, and the use amount of acid acetone is reduced by half compared with the method for extracting heme by an acid acetone method; the reverse osmosis membrane used is a double-layer membrane of a polyamide composite reverse osmosis membrane and a high molecular polymer chitosan reverse osmosis membrane, the separation of polypeptide in the solution is enhanced through the action of chitosan, the purity of the obtained heme is improved, the removal rate of foreign matters in a system is improved through a double-layer membrane structure, and the chlorine resistance, oxidation resistance and pollution resistance of the membrane are improved.
Description
Technical Field
The invention relates to a method for producing heme, in particular to a method for preparing high-purity heme by utilizing a reverse osmosis membrane concentration technology.
Background art:
【1】 With the improvement of living standard of people, the demand of meat food is increasing day by day, the slaughter quantity of livestock and poultry is increasing, and simultaneously, a large amount of blood is generated, if the blood can not be utilized in time, the blood is not only waste of resources, but also causes pollution to the environment.
【2】 According to statistics, 60% of people in the society suffer from anemia, heme can be used as organic iron, can be well absorbed by human bodies, has no side effect, is a good iron-supplementing medicine, and has a good treatment effect on iron-deficiency anemia. The heme can be used as food additive, and can be modified to obtain pigment as edible pigment. The production of heme is therefore of increasing interest.
【3】 The existing methods for extracting heme comprise glacial acetic acid (Zhongguang: research on extracting heme by utilizing glacial acetic acid [ J ]. food science 2004, 20 (4): 90-95), acid acetone (Songyenarmy, etc.: new technology research on extracting and purifying heme [ J ] Guangzhou food industry science and technology, 2004, 20 (1): 6-11) and enzymolysis (Yanxinhong, summer water, enzymatic hydrolysis of hemoglobin to prepare heme peptide [ J ]. food and machinery, 2005(3): 314-:
firstly, a large amount of organic solvent is consumed and is difficult to recover, so that the production cost is increased and the environment is polluted;
secondly, the fresh blood is not easy to store and transport, and the production cost for preparing the heme by using the fresh blood as the raw material is higher. ③ the purity and the yield of the prepared heme are lower.
Disclosure of Invention
In order to solve the problems, the invention discloses a method for extracting heme from blood powder by using a reverse osmosis membrane, which specifically adopts the following technical scheme:
(1) adding 10-20 times of purified water into animal blood powder, and stirring for 15-30 to dissolve the blood powder completely.
(2) Slowly adding 0.5-2 times of protein precipitant into the blood powder solution, stirring for 0.5-1h, and precipitating completely, wherein the protein precipitant is acetone and water 1: 3-1: 5, and (c) a mixed solution.
(3) The solution was centrifuged at 3000r/min to collect the red precipitate. Acetone and water for precipitation 1: 3-1: 5, stirring and washing for 3-5 times, wherein the acetone aqueous solution used each time is 2-3 times of the precipitation volume, and after washing, performing suction filtration to collect precipitates.
(4) Adding acidic acetone solution with pH less than 3 and with precipitation mass of 3-5 times into the precipitate, stirring and extracting for 0.5-1h, vacuum filtering, collecting filtrate, washing the filtrate with acidic acetone for 2-3 times, and mixing the filtrates.
(5) Filtering the filtrate with reverse osmosis membrane concentrating device to obtain concentrated heme crude product, and oven drying to obtain high purity heme.
In a further embodiment of the present invention, the reverse osmosis membrane comprises two membranes, wherein the first membrane is a chitosan reverse osmosis membrane and the second membrane is a polyamide reverse osmosis membrane.
As a further scheme of the invention, the preparation process of the chitosan reverse osmosis membrane comprises the following steps:
(1) 2g to 10g of chitosan is put into a 100ml beaker, added with 3 percent to 6 percent aqueous acetone solution of acetic acid (acetone: water = 2: 4-8) and stirred fully for 0.5 to 1 hour.
(2) And adding 1-5g of triethylene glycol into the uniformly mixed solution, fixing the volume to 100mL, adding a cross-linking agent with the mass of 0.02 time of that of the chitosan, and stirring for 0.5-2h by using a stirrer. The cross-linking agent is one of glutaraldehyde, adipic dialdehyde, adipic acid and diepoxy propane.
(3) Pouring the mixed liquid on a clean glass plate, flattening the film by using a glass rod, placing the film in air for drying, and soaking the film into 2-5% diluted alkali solidification liquid of 278K for 0.5-2h after the fluidity is lost.
(4) Finally, the membrane was soaked with acetone until it was separated from the glass plate, washed clean with deionized water and stored in a moist environment.
As a further scheme of the invention, the preparation scheme of the polyamide reverse osmosis composite membrane comprises the following steps:
(1) preparing m-phenylenediamine aqueous solution with the concentration of 1.7-3.0%;
(2) soaking a polysulfone ultrafiltration membrane (with a film forming concentration of 16%) with PET non-woven fabric as a liner in the m-phenylenediamine aqueous solution in the step (1) for 1-5min to form an aqueous phase liquid layer on the surface of the porous base membrane to obtain a wet membrane;
(3) contacting the wet film with naphtha organic solution containing 0.02% -0.04% of trimesoyl chloride for interfacial polycondensation reaction for 1-5 min;
(4) drying in an oven at 110 ℃ for 2-10 minutes;
(5) and washing with clean water for 10 minutes to obtain the polyamide reverse osmosis membrane.
As a further scheme of the invention, the cross-linked chitosan reverse osmosis membrane and the polyamide composite reverse osmosis membrane are used by simultaneously loading into a reverse osmosis device, wherein the chitosan reverse osmosis membrane is a first layer, and the polyamide composite reverse osmosis membrane is a second layer.
The technical scheme provided by the invention has the beneficial effects that:
(1) the invention uses the animal blood powder as the raw material to extract the heme, and the dried blood powder has long shelf life, is convenient for production and utilization, and reduces the environmental pollution.
(2) The method uses the protein precipitator to accelerate the separation of the protein, thereby reducing the usage amount of the acidic acetone, reducing the usage amount of the acidic acetone by half compared with the method for preparing the heme by the acidic acetone method, and reducing the environmental pollution because the acetone used in the experimental process is filtered out by a reverse osmosis membrane and has high recoverability.
(3) The invention uses the cross-linked chitosan reverse osmosis membrane, and the hydroxyl and the amino contained in the molecular structure of the cross-linked chitosan can form hydrogen bond action with O and N atoms in polypeptide molecules, thereby enhancing the separation of the polypeptide in the solution and improving the purity of the obtained heme.
(4) The invention uses the double-layer membrane structure to greatly improve the removal rate of foreign matters in the solvent in reverse osmosis and improve the chlorine resistance, oxidation resistance and pollution resistance of the membrane. And the double-layer reverse osmosis membrane structure is suitable for solution purification of various systems.
Detailed description of the invention
The present invention will be further described below by way of specific examples.
The reverse osmosis concentration devices used in the following examples are hpml1812-2 reverse osmosis concentrators from Jilin sea technology, Inc. The operating parameters were: pressure: 80 PSI (temperature of 25 deg.C)
And the filtering flow rate is 3.5L/h.
Example 1:
preparation of acidic acetone heme solution:
(1) adding 15 times volume of purified water into 1kg of pig blood powder, and stirring for 30min to dissolve the blood powder completely.
(2) Slowly adding a 1.5-time volume of a mixed solution of acetone and water in a ratio of 1:5 into the blood powder solution, and fully stirring for 1h to fully precipitate the blood powder solution.
(3) The solution was centrifuged at 3000r/min to collect the red precipitate. And stirring and washing the precipitate for 3 times by using a mixed solution of acetone and water in a ratio of 1:5, wherein the acetone aqueous solution used in each time is 2 times of the volume of the precipitate, and filtering and collecting the precipitate after washing.
(4) Adding 2L of acidic acetone solution with pH of 3 into the precipitate, stirring and extracting for 0.5h, vacuum filtering, collecting filtrate, washing the filtrate with acidic acetone for 3 times, and mixing the filtrates to obtain acidic acetone heme solution.
Example 2:
the method comprises the following steps: preparing a cross-linked chitosan reverse osmosis membrane:
(1) 8g of chitosan was put in a 100ml beaker, and 3% acetic acid in acetone aqueous solution (acetone: water = 2: 5) was added thereto, followed by well stirring for 0.5 h.
(2) Adding 3g of triethylene glycol into the uniformly mixed solution, fixing the volume to 100mL, adding glutaraldehyde with the mass of 0.02 time of that of the chitosan, and stirring for 1 hour by using a stirrer.
(3) Pouring the mixed liquid on a clean glass plate, flattening the film by using a glass rod, placing the film in air for drying, and soaking the film in 3% diluted alkali solidification liquid of 278K for 2 hours after the fluidity is lost.
(4) Finally, the membrane was soaked with acetone until it was separated from the glass plate, washed clean with deionized water and stored in a moist environment.
Step two: preparing a polyamide composite reverse osmosis membrane:
(1) preparing m-phenylenediamine aqueous solution with the concentration of 2 percent;
(2) soaking a polysulfone ultrafiltration membrane (with a film forming concentration of 16%) with PET non-woven fabric as a liner in the m-phenylenediamine aqueous solution in the step (1) for 1min to form an aqueous phase liquid layer on the surface of the porous base membrane to prepare a wet membrane;
(3) contacting the wet film with naphtha organic solution containing 0.03% of trimesoyl chloride for interfacial polycondensation reaction for 5 min;
(4) oven drying at 110 deg.C for 10 min;
(5) and washing with clean water for 10 minutes to obtain the polyamide reverse osmosis membrane.
Step three: and (2) loading the cross-linked chitosan reverse osmosis membrane and the polyamide composite reverse osmosis membrane into a reverse osmosis concentration device, concentrating 100g of filtrate obtained in example 1 by using the reverse osmosis device, and drying to obtain 1.05g of finished heme product with the purity of 98.22% and the iron element content of 8.36%.
Example 3:
the method comprises the following steps: preparing a cross-linked chitosan reverse osmosis membrane:
(1) 6g of chitosan was put in a 100ml beaker, and added to a 4% aqueous acetone solution of acetic acid (acetone: water = 2: 8) and stirred well for 0.5 h.
(2) Adding 2g of triethylene glycol into the uniformly mixed solution, fixing the volume to 100mL, adding glutaraldehyde with the mass of 0.02 time of that of the chitosan, and stirring for 1 hour by using a stirrer.
(3) Pouring the mixed liquid on a clean glass plate, flattening the film by using a glass rod, placing the film in air for drying, and soaking the film in 3% diluted alkali solidification liquid of 278K for 2 hours after the fluidity is lost.
(4) Finally, the membrane was soaked with acetone until it was separated from the glass plate, washed clean with deionized water and stored in a moist environment.
Step two: preparing a polyamide composite reverse osmosis membrane:
(1) preparing a m-phenylenediamine aqueous solution with the concentration of 1.5 percent;
(2) soaking a polysulfone ultrafiltration membrane (with the film forming concentration of 16%) with PET non-woven fabric as a liner in the m-phenylenediamine aqueous solution in the step (1) for 3min to form an aqueous phase liquid layer on the surface of the porous base membrane to prepare a wet membrane;
(3) contacting the wet film with naphtha organic solution containing 0.04% of trimesoyl chloride to carry out interfacial polycondensation reaction for 3 min;
(4) oven drying at 110 deg.C for 10 min;
(5) and washing with clean water for 10 minutes to obtain the polyamide reverse osmosis membrane.
Step three: and (2) loading the cross-linked chitosan reverse osmosis membrane and the polyamide composite reverse osmosis membrane into a reverse osmosis concentration device, concentrating 100g of filtrate obtained in example 1 by using the reverse osmosis device, and drying to obtain 1.15g of finished heme product with the purity of 98.14% and the iron element content of 8.34%.
Example 4:
the method comprises the following steps: preparing a cross-linked chitosan reverse osmosis membrane:
(1) 10g of chitosan was put in a 100ml beaker, and 3% acetic acid in acetone aqueous solution (acetone: water = 2: 5) was added thereto, followed by well stirring for 0.5 h.
(2) Adding 4g of triethylene glycol into the uniformly mixed solution, fixing the volume to 100mL, adding glutaraldehyde with the mass of 0.02 time of that of the chitosan, and stirring for 1.5 hours by using a stirrer.
(3) Pouring the mixed liquid on a clean glass plate, flattening the film by using a glass rod, placing the film in air for drying, and soaking the film in 3% diluted alkali solidification liquid of 278K for 2 hours after the fluidity is lost.
(4) Finally, the membrane was soaked with acetone until it was separated from the glass plate, washed clean with deionized water and stored in a moist environment.
Step two: preparing a polyamide composite reverse osmosis membrane:
(1) preparing a 3% m-phenylenediamine aqueous solution;
(2) soaking a polysulfone ultrafiltration membrane (with the film forming concentration of 16%) with PET non-woven fabric as a liner in the m-phenylenediamine aqueous solution in the step (1) for 3min to form an aqueous phase liquid layer on the surface of the porous base membrane to prepare a wet membrane;
(3) contacting the wet film with naphtha organic solution containing 0.03% of trimesoyl chloride to carry out interfacial polycondensation reaction for 5 minutes;
(4) oven drying at 110 deg.C for 5 min;
(5) and washing with clean water for 10 minutes to obtain the polyamide reverse osmosis membrane.
Step three: and (2) loading the cross-linked chitosan reverse osmosis membrane and the polyamide composite reverse osmosis membrane into a reverse osmosis concentration device, concentrating 100g of filtrate obtained in example 1 by using the reverse osmosis device, and drying to obtain 1.27g of finished heme product with the purity of 97.93% and the iron element content of 8.28%.
Example 5:
the method comprises the following steps: preparing a cross-linked chitosan reverse osmosis membrane:
(1) in a 100ml beaker, 7.5g of chitosan was added to a 3% aqueous solution of acetic acid in acetone (acetone: water = 2: 4) and stirred well for 0.5 h.
(2) Adding 3.5g of triethylene glycol into the uniformly mixed solution, fixing the volume to 100mL, adding glutaraldehyde with the mass of 0.02 time of that of the chitosan, and stirring for 1.5 hours by using a stirrer.
(3) Pouring the mixed liquid on a clean glass plate, flattening the film by using a glass rod, placing the film in air for drying, and soaking the film in 3% diluted alkali solidification liquid of 278K for 2 hours after the fluidity is lost.
(4) Finally, the membrane was soaked with acetone until it was separated from the glass plate, washed clean with deionized water and stored in a moist environment.
Step two: preparing a polyamide composite reverse osmosis membrane:
(1) preparing m-phenylenediamine aqueous solution with the concentration of 2 percent;
(2) soaking a polysulfone ultrafiltration membrane (with the film forming concentration of 16%) with PET non-woven fabric as a liner in the m-phenylenediamine aqueous solution in the step (1) for 3min to form an aqueous phase liquid layer on the surface of the porous base membrane to prepare a wet membrane;
(3) contacting the wet film with naphtha organic solution containing 0.03% of trimesoyl chloride to carry out interfacial polycondensation reaction for 3 minutes;
(4) oven drying at 110 deg.C for 5 min;
(5) and washing with clean water for 10 minutes to obtain the polyamide reverse osmosis membrane.
Step three: and (2) loading the cross-linked chitosan reverse osmosis membrane and the polyamide composite reverse osmosis membrane into a reverse osmosis concentration device, concentrating 100g of filtrate obtained in example 1 by using the reverse osmosis device, and drying to obtain 1.35g of finished heme product with the purity of 98.31% and the iron element content of 8.38%.
Example 6:
the method comprises the following steps: preparing a cross-linked chitosan reverse osmosis membrane:
(1) 9g of chitosan was put in a 100ml beaker, and 3% acetic acid in acetone aqueous solution (acetone: water = 2: 5) was added thereto, followed by well stirring for 0.5 h.
(2) Adding 4.25g of triethylene glycol into the uniformly mixed solution, fixing the volume to 100mL, adding glutaraldehyde with the mass of 0.02 time of that of the chitosan, and stirring for 1.5 hours by using a stirrer.
(3) Pouring the mixed liquid on a clean glass plate, flattening the film by using a glass rod, placing the film in air for drying, and soaking the film in 278K 4% diluted alkali solidification liquid for 2 hours after the fluidity is lost.
(4) Finally, the membrane was soaked with acetone until it was separated from the glass plate, washed clean with deionized water and stored in a moist environment.
Step two: preparing a polyamide composite reverse osmosis membrane:
(1) preparing m-phenylenediamine aqueous solution with the concentration of 2.5 percent;
(2) soaking a polysulfone ultrafiltration membrane (with the film forming concentration of 16%) with PET non-woven fabric as a liner in the m-phenylenediamine aqueous solution in the step (1) for 3min to form an aqueous phase liquid layer on the surface of the porous base membrane to prepare a wet membrane;
(3) contacting the wet film with naphtha organic solution containing 0.03% of trimesoyl chloride to carry out interfacial polycondensation reaction for 4 minutes;
(4) oven drying at 110 deg.C for 5 min;
(6) and washing with clean water for 10 minutes to obtain the polyamide reverse osmosis membrane.
Step three: and (2) loading the cross-linked chitosan reverse osmosis membrane and the polyamide composite reverse osmosis membrane into a reverse osmosis concentration device, concentrating 100g of filtrate obtained in example 1 by using the reverse osmosis device, and drying to obtain 1.27g of finished heme product with purity of 97.96% and iron element content of 8.30%.
Example seven:
the acidic acetone method was used to extract hemoglobin from the solution (100 g) prepared in example 1 according to the literature (Songyun et al: New technology for extraction and purification of hemoglobin [ J ]. Guangzhou food industry science and technology, 2004, 20 (1): 6-11). The method comprises the following steps: and (3) taking 100ml of the filtrate in the example 1, adjusting the pH value to 6-7 by using 1.0mol/l NaOH solution, separating out a precipitate, and centrifuging for 10min at 3000r/min to obtain the precipitate. Dissolving the precipitate with 30ml of 0.1mol/l NaOH solution, adjusting the pH value to 5-6, centrifuging at 3000r/min for 10min, and finally washing the precipitate with distilled water to neutrality to obtain 0.87g of heme product with purity of 93.14% and iron content of 7.89%.
Example eight:
100g of the solution prepared in example 1 were worked up according to the seventh protocol of example, comprising the steps of: the pH of the filtrate in the example 1 is adjusted to 6-7 by using 1.0mol/l NaOH solution, precipitate is separated out, and the precipitate is separated by centrifuging at 3000r/min for 20 min. Dissolving the precipitate with 25ml of 0.2mol/l NaOH solution, adjusting the pH value to 6-7, centrifuging at 3000r/min for 20min, and finally washing the precipitate with distilled water to neutrality to obtain 0.86g of heme product, wherein the purity is 93.07%, and the content of iron element is 7.86%
The purity, yield and iron content of heme in each of the above examples were measured as follows:
(1) and (3) measuring the content of heme:
the scheme for measuring the content of the heme in the patent is measured by using a high performance liquid chromatography (Cynanchum, Xiandaqiang. HPLC for measuring the content of the hemin in Shuyue capsules. Xinjiang traditional Chinese medicine, 2005,23(6): 6-7), and the specific scheme is as follows:
selection of a mobile phase:
acetonitrile, methanol and water are respectively used as mobile phases, and the influence of the flow of the components on the separation effect of the heme is examined when the flow rate is 0.6mL/min and the column temperature is 30 ℃.
② drawing of HPLC standard curve
20.0mg of heme standard dried to constant weight at 105 ℃ is precisely weighed. Placing the mixture into a 100m1 volumetric flask, adding 0.1mol/L NaOH solution, diluting to the scale, and shaking up to obtain the product. Accurately measuring l.0, 2.0, 3.0, 4.0 and 5.0mL of heme standard solution, using 0.1mol/L NaOH solution to be fixed in a 100mL volumetric flask, and shaking up. Passing the prepared solution through a 0.22um microfiltration membrane, performing ultrasonic degassing, and performing machine detection.
Measuring the content of the heme sample:
taking a sample dried to constant weight at 105 ℃, and grinding into fine powder; an appropriate amount of the fine powder was weighed and placed in a 100mL volumetric flask. Dissolving with 0.1mol/L sodium hydroxide solution, diluting to scale, shaking, filtering, placing 10m1 filtrate in 100m1 volumetric flask, diluting with 0.1mol/L sodium hydroxide solution to scale, shaking, passing through membrane, degassing, and detecting on machine. Substituting the obtained value into a regression equation to calculate the content.
Purity = mass of heme x 100%/mass of product obtained
(2) Heme yield = (mass of produced heme/mass of blood powder used) × 100%
(3) Determination of iron ion content the determination was carried out using the method in the literature (evaluation of two determination methods of iron ion content in Yuan, Yuan. heme. Histra pharmaceutical industry, 2005,17(4): 63-65):
phenanthroline colorimetry. First, a standard curve is drawn. Then accurately weighing a proper amount of the sample in a 100mL beaker, adding 2mL of concentrated hydrochloric acid, and after completely dissolving, fixing the volume with distilled water. 10mL of the solution was added to a separatory funnel, and 1mL of 1:1 sulfuric acid solution and 2mL of 20% potassium thiocyanate solution were added and mixed. Then 30mL of anhydrous ether is added, ferric thiocyanate formed by ferric iron is extracted by shaking for a plurality of times until the ether layer is colorless, and the ether layer is removed. Transferring the residual water layer into a volumetric flask for constant volume. The absorbance is measured, and the ferrous content is calculated by a standard curve.
The heme yields, purities, iron contents of examples 2-8 were compared:
| item | Example 2 | Example 3 | Example 4 | Example 5 | Example 6 | Example 7 | Example 8 |
| Purity% | 98.22 | 98.14 | 97.93 | 98.31 | 97.96 | 93.14 | 93.07 |
| Yield% | 2.1 | 2.30 | 2.54 | 2.7 | 2.54 | 1.74 | 1.72 |
| Iron element content% | 8.36 | 8.34 | 8.28 | 8.38 | 8.30 | 7.89 | 7.86 |
Examples 2-6 use reverse osmosis membrane concentration to extract heme, and examples 7-8 use acid acetone method to extract heme, and it can be seen from the data that the method and process for extracting heme from blood powder using reverse osmosis membrane according to the present invention are significantly improved from aspects of heme yield, purity, and iron element content compared with the conventional process.
Claims (13)
1. The invention discloses a method for extracting heme from blood powder by using reverse osmosis technology, which comprises the steps of dissolving animal blood powder, particularly pig blood powder, by using purified water, adding a protein precipitator into a mixed solution, collecting precipitates after filtration, adding acidic acetone into the precipitates, performing suction filtration after dissolution, collecting filtrate, concentrating the filtrate by using a modified reverse osmosis membrane to obtain a concentrated heme crude product, and drying to obtain high-purity heme.
2. The method of claim 1, wherein the raw material is animal blood powder selected from one of pig blood powder, duck blood powder, and cattle blood powder.
3. The method of claim 1, wherein the amount of purified water is 10-20 times the mass of the dried blood.
4. The method according to claim 1, wherein the protein precipitant is selected from ethanol, methanol, acetone, dimethylformamide, dimethyl sulfoxide, acetonitrile, 2-methyl-2, 4-pentanediol, saturated ammonium sulfate, and 33% sodium chloride salting-out solution.
5. The process according to claims 1 to 4, wherein the protein precipitant used is acetone and water 1: 3-1: 5, and (c) a mixed solution.
6. A process according to claims 1 to 5, wherein the amount of the acetone/water mixture of the protein precipitant is 0.5 to 2 times the amount of the purified water-soluble blood powder solution.
7. The process according to claim 1, wherein the acidic acetone used has a pH of less than 3.
8. The method according to claims 1 to 7, characterized in that the amount of acidic acetone used is 3-5 times the amount of precipitate obtained by filtration with the addition of protein precipitant.
9. The method of claim 1, wherein the reverse osmosis membrane used comprises two membranes, the first membrane being a chitosan reverse osmosis membrane and the second membrane being a polyamide composite reverse osmosis membrane.
10. The method of claims 1 to 9, wherein the reverse osmosis membrane is prepared by placing 2g to 10g of chitosan in a 100ml beaker, adding 3% -6% acetic acid in acetone water solution (acetone: water = 2: 4-8), mixing well, adding 1-5g triethylene glycol, fixing the volume to 100mL scale, adding a certain amount of cross-linking agent, stirring with a stirrer for 1-2h, pouring the mixed liquid on a clean glass plate, and flattening the membrane by using a glass rod, drying the membrane in the air, soaking the membrane into 278K 2-5% diluted alkali solidification liquid for 0.5-2h after the fluidity is lost, soaking the membrane by using acetone until the membrane is separated from a glass plate, cleaning the membrane by using deionized water, storing the membrane in a humid environment, and putting the membrane into a reverse osmosis concentration device when the membrane is used.
11. The method as claimed in claims 1 to 9, wherein the cross-linking agent used by the chitosan reverse osmosis membrane is one of glutaraldehyde, adipic dialdehyde, adipic acid and diepoxy propane.
12. The method of claims 1-9 wherein the second membrane is a conventional polyamide reverse osmosis composite membrane.
13. The method as claimed in claim 1, wherein a protein precipitant is used to accelerate the separation of protein, and the amount of acidic acetone used is reduced by half compared with the acidic acetone method for extracting hemoglobin; the reverse osmosis membrane used is a double-layer membrane of a polyamide composite reverse osmosis membrane and a high molecular polymer chitosan reverse osmosis membrane, the separation of polypeptide in the solution is enhanced through the action of chitosan, the purity of the obtained heme is improved, the removal rate of foreign matters in the system is improved through a double-parameter membrane structure, and the chlorine resistance, oxidation resistance and pollution resistance of the membrane are improved.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110299597.4A CN112999897A (en) | 2021-03-22 | 2021-03-22 | Method and process for extracting heme from blood meal by using reverse osmosis membrane |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110299597.4A CN112999897A (en) | 2021-03-22 | 2021-03-22 | Method and process for extracting heme from blood meal by using reverse osmosis membrane |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112999897A true CN112999897A (en) | 2021-06-22 |
Family
ID=76403806
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110299597.4A Withdrawn CN112999897A (en) | 2021-03-22 | 2021-03-22 | Method and process for extracting heme from blood meal by using reverse osmosis membrane |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112999897A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115475525A (en) * | 2022-10-06 | 2022-12-16 | 中国矿业大学 | Method for purifying biological dust suppressant by two-stage ultrafiltration |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101085777A (en) * | 2007-07-10 | 2007-12-12 | 青岛波尔旺肉业股份有限公司 | Method for producing sheep blood hemachrome |
| KR100925352B1 (en) * | 2008-08-29 | 2009-11-09 | 주식회사 에이.비.아이 | A method for preparing heme-iron polypeptide polymer having high solubility and absorption rate and heme-iron polypeptide polymer prepared from this method |
| CN101941975A (en) * | 2010-08-19 | 2011-01-12 | 四川省德阳市生化制品有限公司 | Method for preparing heme from animal blood corpuscle powder serving as raw material |
| CN102219788A (en) * | 2011-05-10 | 2011-10-19 | 苏州瑞蓝博中药技术开发有限公司 | Method for extracting heme from blood of wild yak |
| CN106554979A (en) * | 2016-11-18 | 2017-04-05 | 安徽菁硕科技有限公司 | A kind of method of the production haemachrome that combined with acid acetone extraction by enzymatic isolation method |
-
2021
- 2021-03-22 CN CN202110299597.4A patent/CN112999897A/en not_active Withdrawn
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101085777A (en) * | 2007-07-10 | 2007-12-12 | 青岛波尔旺肉业股份有限公司 | Method for producing sheep blood hemachrome |
| KR100925352B1 (en) * | 2008-08-29 | 2009-11-09 | 주식회사 에이.비.아이 | A method for preparing heme-iron polypeptide polymer having high solubility and absorption rate and heme-iron polypeptide polymer prepared from this method |
| CN101941975A (en) * | 2010-08-19 | 2011-01-12 | 四川省德阳市生化制品有限公司 | Method for preparing heme from animal blood corpuscle powder serving as raw material |
| CN102219788A (en) * | 2011-05-10 | 2011-10-19 | 苏州瑞蓝博中药技术开发有限公司 | Method for extracting heme from blood of wild yak |
| CN106554979A (en) * | 2016-11-18 | 2017-04-05 | 安徽菁硕科技有限公司 | A kind of method of the production haemachrome that combined with acid acetone extraction by enzymatic isolation method |
Non-Patent Citations (2)
| Title |
|---|
| 蔡君等: "酶法结合膜技术提取血红素的工艺研究", 《食品科技》 * |
| 陈兴凡等: "壳聚糖反渗透膜的研制", 《上海师范大学学报(自然科学版)》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115475525A (en) * | 2022-10-06 | 2022-12-16 | 中国矿业大学 | Method for purifying biological dust suppressant by two-stage ultrafiltration |
| CN115475525B (en) * | 2022-10-06 | 2024-03-05 | 中国矿业大学 | Method for purifying biological dust suppressant by double-stage ultrafiltration |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104342040B (en) | A kind of high glue melts the preparation method of temperature fish scale gelatin | |
| CN106397630B (en) | A method of Sodium Hyaluronate is extracted using membrane separation technique | |
| CN104892710B (en) | A kind of method for purifying reduced form β NADHs | |
| CN102351956B (en) | Extraction method of morindea officinalis polysaccharide | |
| CN101029077B (en) | Method for purifying gene-recombinant insulin precursor | |
| CN103627768A (en) | Fish skin collagen bioactive small peptide and preparation method thereof | |
| CN112999897A (en) | Method and process for extracting heme from blood meal by using reverse osmosis membrane | |
| CN105648008A (en) | Feeding silk antibacterial peptide preparation and preparation method thereof | |
| CN111359594A (en) | Boric acid adsorption material and preparation method thereof | |
| CN103880945A (en) | Method for preparing high-purity thymalfasin | |
| CN110144376B (en) | Nano-scale collagen peptide and preparation method thereof | |
| CN101941975B (en) | Method for preparing heme from animal blood corpuscle powder serving as raw material | |
| CN107619411B (en) | Heme extraction method | |
| CN104774827A (en) | Method for preparing alginate lyase from abalone internal organs | |
| CN104530260A (en) | Method for co-producing high-purity heparin sodium and dermatan sulfate from pig lungs | |
| CN103059159A (en) | Process for extracting mannan from beer yeast powder | |
| CN102816224B (en) | Hair keratin solution for spinning, preparation method and application thereof | |
| CN112646055A (en) | Preparation method of low-molecular-weight hyaluronic acid | |
| CN111617636B (en) | Plant extraction and purification process | |
| CN104558251B (en) | A kind of preparation method of liquaemin | |
| CN114807284A (en) | Preparation method of high-purity small-molecule fish skin collagen peptide | |
| CN109384861A (en) | A kind of method of heparin sodium pulp thickening dermatan sulfate | |
| CN102391363B (en) | Method for extracting enramycin | |
| CN105732797A (en) | Purification method for urinary gonadotropin | |
| CN112029015A (en) | Production and purification process of high-purity low-molecular-weight heparin sodium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20210622 |
|
| WW01 | Invention patent application withdrawn after publication |