CN1322761A - Animal hematoglobin and its extraction process and use - Google Patents
Animal hematoglobin and its extraction process and use Download PDFInfo
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- CN1322761A CN1322761A CN01114651A CN01114651A CN1322761A CN 1322761 A CN1322761 A CN 1322761A CN 01114651 A CN01114651 A CN 01114651A CN 01114651 A CN01114651 A CN 01114651A CN 1322761 A CN1322761 A CN 1322761A
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- FWZTTZUKDVJDCM-CEJAUHOTSA-M disodium;(2r,3r,4s,5s,6r)-2-[(2s,3s,4s,5r)-3,4-dihydroxy-2,5-bis(hydroxymethyl)oxolan-2-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol;iron(3+);oxygen(2-);hydroxide;trihydrate Chemical compound O.O.O.[OH-].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[Na+].[Na+].[Fe+3].[Fe+3].[Fe+3].[Fe+3].[Fe+3].O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 FWZTTZUKDVJDCM-CEJAUHOTSA-M 0.000 description 1
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Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The electrostatic adsorption and separation process of extracting hematoglobin from animal erythrocyte to replace traditional centrifugal separation process includes addition of proper amount of Ca ion at low temp environment of 2-8 deg.c, chromatographic separation of erythrocyte from blood plasma, extracting, erythrocyte in the lower layer while eliminating blood plasma in the upper layer and acid hydrolysis of eryrhrocyte to extract hematoglobin. The hematoglobin is used as main material or additive in preparing medicine and health food for preventing and treating anaemia.
Description
The present invention relates to a kind of animal blood haemoglobin and extracting method thereof and purposes.Be meant the purposes of from animal red blood corpuscle (red corpuscle), extracting oxyphorase and this oxyphorase with special method.Belong to medicine and health product technology field
Anaemia is a kind of common disease, the all weary color of light disease anaemia patient face lip and mucous membrane, cardinal symptom is a pale complexion: significant anaemia patient is except that the symptom with patient with slight symptoms, xerasia also appears, nail is a spoon coelonychia, and liver, splenic lymph nodes enlargement can appear, appetite stimulator, symptom such as palpitaition, weak, fidgety, lassitude, absent minded, poor memory, dizziness, tinnitus.In the prior art, the methods of treatment of anaemia is had two kinds, a kind of is to take food and the vitamins C that is rich in irony; Another kind is pharmacological agent, is divided into oral ferrous sulfate, glucose is ferrous and intravenous injection saccharated iron oxide or glucoferrum.Wherein oral ferrous sulfate or ammonium ferric citrate are bigger to gastric stimulation, symptoms such as nausea,vomiting,diarrhea can appear behind the clothes, though take the stimulation that can reduce chalybeate after meal, but because food waters down the alienization that hydrochloric acid in gastric juice lowers iron, phosphide in the food combines with iron and generates undissolved phosphoric acid salt, thereby influence the absorption of human body to iron, unsatisfactory curative effect; Intravenous therapy only is applicable to following several situation: severe reaction all takes place in (1) oral various iron, and it is still invalid to change dosage and medicine time.(2) gastrointestinal tract disease influences the absorption of iron.(3) secondary hypoferric anemia after the gastrointestinal surgery.(4) use oral chalybeate treatment blood picture and do not have the changer.Though blood transfusion can make the patient come back to life, blood transfusion is to be lower than the critical patient in 3% when gram at oxyphorase just to use on one's body, and blood transfusion can only solve temporary anaemia, can not thoroughly effect a radical cure anaemia.
Discover, sophisticated red blood corpuscle (red corpuscle) contains abundant oxyphorase in the animal blood, its sophisticated red corpuscle is more smaller than human red corpuscle, major diameter is 6.2～6.5 microns, shaped flat, two sides depression, disk-shaped, central authorities' minor axis is 1.6～2.0 microns, extracts the oxyphorase a part oxyphorase that obtains and contain 4 iron porphyrins from red corpuscle, and these iron porphyrins are the natural medicine of treatment anaemia and the protective foods of enriching blood.Therefore, extracting oxyphorase from animal blood, take to the anaemia patient these animal blood haemoglobins, is to replenish the red proteic preferable approach of blood in human body in.
Aforementioned analysis as can be known, oral chalybeate medicine easily causes toxic, and intravenous injection can only be eliminated temporary transient symptom, neither is the ideal methods of treatment therefore.Have only and take in natural oxyphorase in right amount in the food process on the feed, take for a long time and do not have toxic side effects, could effect a radical cure hypoferric anemia.
The classical in the past method of extracting oxyphorase has two kinds: a kind of is to adopt human blood or animal blood to add high-potassium ferricyanide, removes Hbs and Hbc, can obtain cyanmethemoglobin; Another kind is to extract purified chlorination oxyphorase with nonionic washing agent alkali solution (AHD reagent) from blood.But the shortcoming that the oxyphorase that these methods are extracted exists has: (1) complex process, and the cost height causes valuable product; (2) the deleterious chemical residue of Zhi Bei oxyphorase can not only can be used as chemical reagent and use as medicine and food uses.
Though adopted centrifugal separation method open now from the technology that animal plasma extracts oxyphorase, for example: the disclosed a kind of name of Chinese patent communique is called " active iron and production method thereof ", number of patent application is the application for a patent for invention of " 93105261.0 ", a kind of method of extracting oxyphorase from animal plasma is disclosed, be characterized in: get animal plasma and put into whizzer and isolate oxyphorase or serum, serum is put into the electrophoretic separation machine again isolate wherein ionic trace element.This is the technical scheme that a kind of blood plasma that utilizes animal blood partly extracts oxyphorase, though can replenish iron to the patient with the form of food, but have following shortcoming: the raw material that adopt (1) is the blood plasma that is positioned at the upper strata after centrifugation, the content of hemoglobin of this blood plasma is lower, therefore the oxyphorase that extracts is a kind of free hemoglobin, and its purity has only about 60%, oxyphorase recovery rate less than 10%, the cost height, yield poorly, be not suitable for suitability for industrialized production; (2) used antithrombotics, great majority are sodium citrates, this class antithrombotics can not suppress the oxygenizement of oxyphorase, and the oxyphorase of extraction is lost activity; (3) adopt the oxyphorase goods difficulty of the open production of whizzer to give the hygienic standard that reaches national drug and food, do not have DEVELOPMENT PROSPECT.
As everyone knows, blood or be called whole blood.After blood exsomatized, through clot occurring after a while, blood clot retraction can form some clear liquid, promptly is called serum.When getting fresh blood, if add antithrombotics in advance, after blood and antithrombotics mixing, the blood of obtaining is the suspendible shape, does not produce grumeleuse, but through the long period or after centrifugation, supernatant layer and sinking blood cell layer also can appear in this anticoagulation, wherein supernatant layer medically is called blood plasma, and it is that (or claim: red corpuscle), the tunica albuginea that both intersections form is called white cell and platelet layer to red blood corpuscle that the layer that sinks is called blood cell.
The objective of the invention is for a kind of oxyphorase and extracting method and purposes of extracting from animal red blood corpuscle (red corpuscle) is provided.
Purpose of the present invention can reach by taking following measure: a kind of animal blood haemoglobin is characterized in: adopt animal red blood corpuscle (red corpuscle), extract oxyphorase after acid hydrolysis.
The extracting method of foregoing a kind of animal blood haemoglobin is characterized in:
Adopt " electrostatic adhesion partition method " to replace tradition habitual " centrifugal separation ", add proper C a
+Ion also is under 2 ℃～8 ℃ low temperature environments, makes blood plasma and red blood corpuscle (red corpuscle) chromatographic separation, draws and discard upper plasma again, extracts lower floor's red blood corpuscle (red corpuscle), utilizes red blood corpuscle (red corpuscle) to extract oxyphorase after acid hydrolysis.The concrete step is poly-as follows:
1) gets the healthy animal fresh blood of butchering immediately, directly add in the container of the solution of preparing with 1.25% charcoal acid calcium, 0.9%NaCl, 0.2% Calcium Disodium Versenate EDTA.Na (or ethylene dinitrilotetra-acetic acid dipotassium EDTA.K) and aseptic deionized water, the limit edged stirs, left standstill about 1 hour after stopping to stir, the freezer of sending into 2 ℃～8 ℃ then left standstill 12 to 24 hours, through Ca
+Ion electrostatic adhesion, low temperature chromatography, siphon separates blood plasma with red blood corpuscle (red corpuscle), abandon blood plasma, gets red blood corpuscle (red corpuscle);
2) leave standstill in the process positively charged Ca at 2 ℃～8 ℃ freezer
+Ion makes mutually adsorpting aggregation of electronegative red blood corpuscle (red corpuscle), and descends rapidly, makes blood plasma and red blood corpuscle (red corpuscle) layering, and blood plasma is positioned at the upper strata, and red blood corpuscle (red corpuscle) is positioned at lower floor, and the siphon slurry of dehematizing is got red blood corpuscle (red corpuscle);
3) calculate red blood corpuscle (red corpuscle) total amount, add the equivalent aseptic deionized water, the Glacial acetic acid 30% of adding 10% under stirrer fully stirs, continue to add the EDTA.K of 0.1M and 0.5% an amount of sodium sulphite, continue to stir 30 to 60 minutes, make the abundant cracking of red blood corpuscle (red corpuscle) and discharge oxyphorase, 180～200 rev/mins of stirring velocitys;
4) in 3 ℃～5 ℃ freezer, placed 8 to 16 hours, take out back 1M NaHCO
3Potential of hydrogen is adjusted to PH7.0, selects 0.3 μ cellulose acetate film and filter paper plate two times of ultrafiltration, get filtrate, add 1.25% potassium aluminium sulfate, get precipitation with the Cai Shi funnel, concentrating under reduced pressure, vacuum-drying obtains purity and reaches hemoglobin crystal more than 95%.
The extracting method of foregoing a kind of animal blood haemoglobin, its characteristics also have:
The animal that is used to gather blood comprises pig, ox, sheep, horse, chicken, duck, goose, dog, rabbit.
Adopt " electrostatic adhesion partition method " that blood plasma is separated with red blood corpuscle (red corpuscle), abandon blood plasma, get red blood corpuscle (red corpuscle).
Adopt electrostatic adhesion, low temperature chromatography and siphon that blood plasma is separated with red blood corpuscle (red corpuscle), abandon blood plasma, get red blood corpuscle (red corpuscle).
The Ca of electrostatic separation blood plasma and oxyphorase (red corpuscle)
+Ion adopts calcified material, is respectively charcoal acid calcium, calcium hydroxide, calcium lactate or calcium chloride.
The present invention adopts " electrostatic adhesion partition method " to replace tradition habitual " centrifugal separation ", extracts oxyphorase, receives the effect of getting twice the result with half the effort.Its principle is the principle according to the same sex on the physics is repelled each other, there is a natural attraction between the sexes.Because electronegative polarized state and the static membrane potential difference of red blood corpuscle (red corpuscle) in the blood, so under normal circumstances, make the red blood corpuscle that is flat concave surface can not produce aggegation or overlapping, maintain a certain distance between the blood cell, be difficult for natural subsidence, but add proper C a
+Ion (for example charcoal acid calcium) also is under 2 ℃～8 ℃ low temperature environments, owing to changed electrolytical ionic concn in the blood (positive and negative ionization change), therefore red blood corpuscle (red corpuscle) but assemble mutually and rapid subsidence, blood plasma separates naturally with red blood corpuscle (red corpuscle), red blood corpuscle (red corpuscle) is in lower floor, blood plasma floats over the upper strata, with siphon the upper plasma sucking-off is discarded again, takes off a layer red blood corpuscle (red corpuscle) and directly extracts oxyphorase.
The purposes of foregoing a kind of animal blood haemoglobin is characterized in:
Be used for making prevention and treatment anaemia medicine and protective foods, perhaps as the additive of making aforementioned medicine and protective foods.
Described prevention and treatment anaemia medicine and protective foods comprise: oxyphorase oral liquid, oxyphorase medicinal extract, oxyphorase electuary, oxyphorase capsule, oxyphorase tablet; Oxyphorase drink, oxyphorase candy, oxyphorase biscuit, oxyphorase cake, oxyphorase ice cream, oxyphorase snow batch (ice-lolly), the sweet letter of oxyphorase.
Compared with prior art, the present invention has following outstanding advantage:
(1) " centrifugal separation " of the prior art, the red blood corpuscle of separating are chaotic deadlock, therefore, can only utilize the blood plasma that is positioned at the upper strata to extract oxyphorase.The present invention adopts " electrostatic adhesion partition method ", and the layer of red blood cells after the separation is the sequence beds of precipitation, blood plasma and the red blood corpuscle separate easily of rule.The red blood corpuscle (red corpuscle) that utilization is in lower floor extracts oxyphorase, and the oxyphorase purity of extraction can reach more than 95%, and the recovery rate of oxyphorase reaches more than 90%, and cost is low, output is big, is fit to suitability for industrialized production.
(2) the present invention makes blood anticoagulant at collection animal blood, selection and adding EDTA.K or EDTA.Na, both can suppress blood coagulation, be convenient to separated plasma and separation of red blood cells (red corpuscle), the oxygenizement that also can suppress oxyphorase guarantees the active stability of oxyphorase.
(3) the present invention adopts the red blood corpuscle (red corpuscle) of animal blood, work workshop closed under the sanitary condition (GMP) of strict ultra-clean is produced oxyphorase, its finished product meet the food hygienic standard and the international hygiene standard of national regulation, product can export, can be used as medicine and food and use, for prevention and treatment anaemia purposes.
(4) it is simply nontoxic that production of the present invention utilizes the oxyphorase technology of animal bloods such as pig, ox, sheep, horse, chicken, duck, goose, dog, rabbit, and no chemical residue can supply medicinal use and food prepared therefrom, and is safe and effective.
As everyone knows, oxyphorase is a kind of iron-protein, and this iron-protein occupies critical positions in blood, and therefore the supply to iron-protein is extremely important in the activity of life.The intake of child Shi Tie must surpass excretion, otherwise children are easy to iron deficiency, cause anemia.General baby's body iron content is 0.5 gram, adult's 3～5 grams, so children's must be from external 2.5～4.5 gram iron that obtain at least in growth and development process, be 6～16 milligrams of iron of per day needs, women's requirement is more than the male sex again, if normal red corpuscle iron deficiency can influence red corpuscle normal existence in blood circulation, and easily suffers from iron deficiency sexual cell property anaemia.Storage iron in the normal human, if the normovolemia of former storage, when being about as much as three/for the moment of total amount in the whole blood in the body, even if additionally do not replenish chalybeate, blood picture also can recover rapidly normally anaemia can not take place.If but long-term chronic blood loss whenever loses blood 4 milliliters and be roughly equal to 1 milligram of iron, though lose blood every day seldom, the iron blood volume of actual consumption above the several times of normal consumption amount, is accumulated over a long period, causes anaemia naturally.Must replenish iron this moment from external absorption.
Extracting highly purified oxyphorase from animal blood is a kind of albumen of pure natural, and as health care medicine additive or foodstuff additive, the health care medicine of manufacturing or food are pure natural diet articles for use, are easy to be absorbed by the patient with this oxyphorase.Form with food is taken to the patient, replaces traditional medication with dietotherapy, not only can improve the particularly interest of child patient of user, and do not take the toxic side effects of other drug, need not consider the danger of overdose, can effect a radical cure anaemia, have additional nutrients, safe and efficient.
The blood resource of animal pig, ox, sheep, horse, chicken, duck, goose, dog, rabbit etc. is abundant, it is simple to extract blood red egg technical qualification and technology certainly from the red blood corpuscle (red corpuscle) of animal, be easy to grasp and implement, therefore the present invention relates to animal blood haemoglobin and have quality guarantee with the medicine and the protective foods of this oxyphorase manufacturing, with low cost.
Describe the present invention below in conjunction with embodiment:
Embodiment 1:
1) removes ionized water 2500ml, add 0.9%NaCl, 0.2% Calcium Disodium Versenate and 1.25% charcoal acid calcium, be sealed in the container and sterilize, on 4 ℃ cold condition, left standstill at least 2 hours, fresh pig blood is directly injected said vesse, stirring while annotating, is 10000ml until mixing total blood volume, leaves standstill 1 hour again;
2) change in 2 ℃～8 ℃ the freezer and leave standstill, adopt siphon suction to be positioned at the blood plasma on upper strata and discard, get red blood corpuscle (red corpuscle) to 24 hours;
3) described red blood corpuscle (red corpuscle) is sent into the GMP workshop;
4) total amount of metering red blood corpuscle (red corpuscle), add the equivalent aseptic deionized water, fully stir, add 10% Glacial acetic acid 300ml, sodium sulfate 50 grams, EDTA.K 10 grams subsequently, continue to stir 1 hour, make fully cracking and discharge intracellular oxyphorase of red blood corpuscle (red corpuscle), place 4 ℃ freezer 12 hours, take out and use 1M NaHCO
3Hydrogen ion concentration is transferred to PH7.0, select 0.3 μ cellulose acetate film and twice positive press filtration of filter paper plate, get filtrate, add 1.25% potassium aluminium sulfate 125ml, abandon supernatant liquor, get precipitation with the Cai Shi funnel; Thin film concentration is to the medicinal extract sample, and vacuum-drying obtains the red-brown hemoglobin crystal, and purity reaches more than 95%; Measure oxyphorase with cyanmethemoglobin (HiCN) method;
5) in the quantitative packing in GMP workshop, every bag 10mg makes electuary, as natural medicine or healthcare products purposes.
Embodiment 2:
1) removes ionized water 5000ml, add 0.9%NaCl, 0.2% Calcium Disodium Versenate and 1.25% charcoal acid calcium, be sealed in the container and sterilize, on 4 ℃ cold condition, left standstill at least 2 hours, fresh sheep blood is directly injected said vesse, stirring while annotating, is 20000ml until mixing total blood volume, leaves standstill 1 hour again;
2) change in 2 ℃～8 ℃ the freezer and leave standstill, adopt siphon suction to be positioned at the blood plasma on upper strata and discard, get red blood corpuscle (red corpuscle) to 24 hours;
3) described red blood corpuscle (red corpuscle) is sent into the GMP workshop;
4) total amount of metering red blood corpuscle (red corpuscle), add the equivalent aseptic deionized water, fully stir, add 10% Glacial acetic acid 600ml, sodium sulfate 100 grams, EDTA.K 20 grams subsequently, continue to stir 1 hour, make fully cracking and discharge intracellular oxyphorase of red blood corpuscle (red corpuscle), place 4 ℃ freezer 12 hours, take out and use 1M NaHCO
3Hydrogen ion concentration is transferred to PH7.0, select 0.3 μ cellulose acetate film and twice positive press filtration of filter paper plate, get filtrate, add 1.25% potassium aluminium sulfate 250ml, abandon supernatant liquor, get precipitation with the Cai Shi funnel; Thin film concentration is to the medicinal extract sample, and vacuum-drying obtains the red-brown hemoglobin crystal, and purity reaches more than 95%; Measure oxyphorase with cyanmethemoglobin (HiCN) method;
5) quantitatively be pressed into tablet in the GMP workshop, every 0.3 gram is as natural medicine or healthcare products purposes.
Embodiment 3:
1) removes ionized water 3750ml, add 0.9%NaCl, 0.2% Calcium Disodium Versenate and 1.25% charcoal acid calcium, be sealed in the container and sterilize, on 4 ℃ cold condition, left standstill at least 2 hours, fresh bovine blood is directly injected said vesse, stirring while annotating, is 15000ml until mixing total blood volume, leaves standstill 1 hour again;
2) change in 2 ℃～8 ℃ the freezer and spend the night, leave standstill, adopt siphon suction to be positioned at the blood plasma on upper strata and discard, get red blood corpuscle (red corpuscle) to 24 hours;
3) described red blood corpuscle (red corpuscle) is sent into the GMP workshop;
4) total amount of metering red blood corpuscle (red corpuscle), add the equivalent aseptic deionized water, fully stir, add 10% Glacial acetic acid 450ml, sodium sulfate 75 grams, EDTA.K 30 grams subsequently, continue to stir 1 hour, make fully cracking and discharge intracellular oxyphorase of red blood corpuscle (red corpuscle), place 4 ℃ freezer 12 hours, take out and use 1M NaHCO
3Hydrogen ion concentration is transferred to PH7.0, select 0.3 μ cellulose acetate film and twice positive press filtration of filter paper plate, get filtrate, add 1.25% potassium aluminium sulfate 375ml, abandon supernatant liquor, get precipitation with the Cai Shi funnel; Thin film concentration is to the medicinal extract sample, and vacuum-drying obtains the red-brown hemoglobin crystal, and purity reaches more than 95%, measures oxyphorase with cyanmethemoglobin (HiCN) method;
5) add an amount of essence and granulated sugar 300 grams, quantitatively be distributed into oral liquid in the GMP workshop, every 1 gm/10ml is as natural medicine or healthcare products purposes.
Embodiment 4:
1) removes ionized water 7500ml, add 0.9%NaCl, 0.2% Calcium Disodium Versenate and 1.25% charcoal acid calcium, be sealed in the container and sterilize, on 4 ℃ cold condition, left standstill at least 2 hours, fresh horse blood is directly injected said vesse, the limit edged stirs, and is 30000ml until mixing total blood volume, leaves standstill 1 hour again;
2) change over to and cross liquid in 2 ℃～8 ℃ the freezer, leave standstill, adopt siphon suction to be positioned at the blood plasma on upper strata and discard, get red blood corpuscle (red corpuscle) to 24 hours;
3) described red blood corpuscle (red corpuscle) is sent into the GMP workshop;
4) total amount of metering red blood corpuscle (red corpuscle), add the equivalent aseptic deionized water, fully stir, add 10% Glacial acetic acid 900ml, sodium sulfate 150 grams, EDTA.K 30 grams subsequently, continue to stir 1 hour, make fully cracking and discharge intracellular oxyphorase of red blood corpuscle (red corpuscle), place 4 ℃ freezer 12 hours, take out and use 1M NaHCO
3Hydrogen ion concentration is transferred to PH7.0, select 0.3 μ cellulose acetate film and twice positive press filtration of filter paper plate, get filtrate, add 1.25% potassium aluminium sulfate 375ml, abandon supernatant liquor, get precipitation with the Cai Shi funnel; Thin film concentration is to the medicinal extract sample, and vacuum-drying obtains the red-brown hemoglobin crystal, and purity reaches more than 95%; Measure oxyphorase with cyanmethemoglobin (HiCN) method;
5) be processed into micro mist in the GMP workshop, be distributed into the pulvis of every bag 10 grams, as natural medicine and healthcare products and additive, for material outlet to 500 grams.
The characteristics of embodiments of the invention 5 are: the temperature of freezer is 2 ℃～8 ℃, the Ca of electrostatic separation blood plasma and oxyphorase (red corpuscle)
+Ion adopts calcium hydroxide, and all the other are with arbitrary embodiment among the embodiment 1 to embodiment 4.
The characteristics of embodiments of the invention 6 are: the temperature of freezer is 2 ℃～8 ℃, the Ca of electrostatic separation blood plasma and oxyphorase (red corpuscle)
+Ion adopts calcium chloride, and all the other are with arbitrary embodiment among the embodiment 1 to embodiment 4.
The characteristics of embodiments of the invention 7 are: make the abundant cracking of red blood corpuscle and discharge endoerythrocytic oxyphorase, adopt 4% Glacial acetic acid, all the other are with arbitrary among the embodiment 1 to embodiment 4
Embodiment.
The characteristics of embodiments of the invention 8 are: the stirring velocity that makes the abundant cracking of red blood corpuscle and discharge endoerythrocytic oxyphorase is 200 rev/mins, and all the other are with arbitrary embodiment among the embodiment 1 to embodiment 4.
Claims (9)
1. an animal blood haemoglobin is characterized in that: adopt animal red blood corpuscle (red corpuscle), extract oxyphorase after acid hydrolysis.
2. the extracting method of animal blood haemoglobin as claimed in claim 1 is characterized in that: adopt " electrostatic adhesion partition method " to replace tradition habitual " centrifugal separation ", add proper C a
+Ion also is under 2 ℃～8 ℃ low temperature environments, makes blood plasma and red blood corpuscle (red corpuscle) chromatographic separation, draws and discard upper plasma again, extracts lower floor's red blood corpuscle (red corpuscle), utilizes red blood corpuscle (red corpuscle) to extract oxyphorase after acid hydrolysis; The concrete step is poly-as follows:
1) gets the healthy animal fresh blood of butchering immediately, directly add in the container of the solution of preparing with 1.25% charcoal acid calcium, 0.9%NaCl, 0.2% Calcium Disodium Versenate EDTA.Na (or ethylene dinitrilotetra-acetic acid dipotassium EDTA.K) and aseptic deionized water, the limit edged stirs, left standstill about 1 hour after stopping to stir, the freezer of sending into 2 ℃～8 ℃ then left standstill 12 to 24 hours, through Ca
+Ion electrostatic adhesion, low temperature chromatography, siphon separates blood plasma with red blood corpuscle (red corpuscle), abandon blood plasma, gets red blood corpuscle (red corpuscle);
2) leave standstill in the process positively charged Ca at 2 ℃～8 ℃ freezer
+Ion makes mutually adsorpting aggregation of electronegative red blood corpuscle (red corpuscle), and descends rapidly, makes blood plasma and red blood corpuscle (red corpuscle) layering, and blood plasma is positioned at the upper strata, and red blood corpuscle (red corpuscle) is positioned at lower floor, and the siphon slurry of dehematizing is got red blood corpuscle (red corpuscle);
3) calculate red blood corpuscle (red corpuscle) total amount, add the equivalent aseptic deionized water, the Glacial acetic acid 30% of adding 10% under stirrer fully stirs, continue to add the EDTA.K of 0.1M and 0.5% an amount of sodium sulphite, continue to stir 30 to 60 minutes, make the abundant cracking of red blood corpuscle (red corpuscle) and discharge oxyphorase, 180～200 rev/mins of stirring velocitys;
4) in 3 ℃～5 ℃ freezer, placed 8 to 16 hours, take out back 1M NaHCO
3Potential of hydrogen is adjusted to PH7.0, selects 0.3 μ cellulose acetate film and filter paper plate two times of ultrafiltration, get filtrate, add 1.25% potassium aluminium sulfate, get precipitation with the Cai Shi funnel, concentrating under reduced pressure, vacuum-drying obtains purity and reaches hemoglobin crystal more than 95%.
3. the extracting method of a kind of animal blood haemoglobin as claimed in claim 2 is characterized in that: the animal that is used to gather blood comprises pig, ox, sheep, horse, chicken, duck, goose, dog, rabbit.
4. the extracting method of a kind of animal blood haemoglobin as claimed in claim 1 is characterized in that: adopt " electrostatic adhesion partition method " that blood plasma is separated with red blood corpuscle (red corpuscle), abandon blood plasma, get red blood corpuscle (red corpuscle).
5. the extracting method of a kind of animal blood haemoglobin as claimed in claim 1 is characterized in that: adopt electrostatic adhesion, low temperature chromatography and siphon that blood plasma is separated with red blood corpuscle (red corpuscle), abandon blood plasma, get red blood corpuscle (red corpuscle).
6. the extracting method of a kind of animal blood haemoglobin as claimed in claim 2 is characterized in that: the Ca of electrostatic separation blood plasma and red blood corpuscle (red corpuscle)
+Ion adopts calcified material, is respectively charcoal acid calcium, calcium hydroxide, calcium lactate or calcium chloride.
7. the extracting method of a kind of animal blood haemoglobin as claimed in claim 2 is characterized in that: the Ca of electrostatic separation blood plasma and red blood corpuscle (red corpuscle)
+Ion adopts charcoal acid calcium.
8. the purposes of oxyphorase as claimed in claim 1 is characterized in that: be used for making prevention and treatment anaemia medicine and protective foods, perhaps as the additive of making aforementioned medicine and protective foods.
9. the purposes of oxyphorase as claimed in claim 8 is characterized in that: prevention and treatment anaemia medicine and protective foods comprise: oxyphorase oral liquid, oxyphorase medicinal extract, oxyphorase electuary, oxyphorase capsule, oxyphorase tablet; Oxyphorase drink, oxyphorase candy, oxyphorase biscuit, oxyphorase cake, oxyphorase ice cream, oxyphorase snow batch (ice-lolly), oxyphorase ice cream cone.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100363381C (en) * | 2006-01-20 | 2008-01-23 | 南京师范大学 | Method for extracting and purifying metmyoglobin from myocardium |
| CN105092328A (en) * | 2015-07-30 | 2015-11-25 | 厦门宝太生物科技有限公司 | Erythrocyte removing treating fluid and application |
| CN110988366A (en) * | 2019-12-16 | 2020-04-10 | 浙江卫未生物医药科技有限公司 | Reagent and method for rapidly judging sample in-vitro time |
| CN112359014A (en) * | 2020-09-10 | 2021-02-12 | 吉林基蛋生物科技有限公司 | Preparation method of animal peripheral red blood cells |
| WO2021110127A1 (en) * | 2019-12-06 | 2021-06-10 | Cheer Global Limited | Method of treating inflammatory bowel disease |
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2001
- 2001-04-25 CN CN01114651A patent/CN1322761A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100363381C (en) * | 2006-01-20 | 2008-01-23 | 南京师范大学 | Method for extracting and purifying metmyoglobin from myocardium |
| CN105092328A (en) * | 2015-07-30 | 2015-11-25 | 厦门宝太生物科技有限公司 | Erythrocyte removing treating fluid and application |
| WO2021110127A1 (en) * | 2019-12-06 | 2021-06-10 | Cheer Global Limited | Method of treating inflammatory bowel disease |
| CN114786705A (en) * | 2019-12-06 | 2022-07-22 | 环喜有限公司 | Method of treating inflammatory bowel disease |
| US11535655B2 (en) | 2019-12-06 | 2022-12-27 | Cheer Global Limited | Method of treating inflammatory bowel disease |
| CN114786705B (en) * | 2019-12-06 | 2023-05-12 | 环喜有限公司 | Method for treating inflammatory bowel disease |
| CN110988366A (en) * | 2019-12-16 | 2020-04-10 | 浙江卫未生物医药科技有限公司 | Reagent and method for rapidly judging sample in-vitro time |
| CN112359014A (en) * | 2020-09-10 | 2021-02-12 | 吉林基蛋生物科技有限公司 | Preparation method of animal peripheral red blood cells |
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