• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    There is provided an improved method, suitable for commercial use, for separating from a liquid containing blood substances the iron protoporphyrin and dirivatives or decomposition products thereof, the so-called iron component, from globin, characterized in, that the liquid is cooled, preferably to a temperature of from 0.degree. to 20.degree.C, that the pH of the liquid is adjusted to a value lower then 4.5, preferably to a value of from 2,5 to 4.5, that an organic solvent with dehydrating properties is added to said liquid, for example ethanol or mixtures of ethanol with glycol, in an amount liquid, preferably at least 75 per cent of volume, and thereby maintaining the above indicated temperature, that the precipitated iron component is separated from said liquid, preferably by addition of salts, such as potassium chloride or sodium citrate, whereafter the globin is precipitated from the liquid.
    公开号:SU910135A3
    申请号:SU752143704
    申请日:1975-06-13
    公开日:1982-02-28
    发明作者:Геран Сигвард Линдросс Пауль
    申请人:за витель Иностранец Пауль Геран Сигварп Лин росс (Швеци )；
    IPC主号:
    专利说明:

    (54) METHOD OF SEPARATION OF HEMO-IRON FROM GLOBIN
    one
    This invention relates to hematology and concerns the treatment of fluids containing blood constituents.
    HinecreH is a method for the separation of hemoglobin in the blood, including the step-by-step treatment of raS 5 solution containing hemoglobin, salts and organic solvents {.
    However, an honest method does not provide a high yield and preserve the native properties of globin.
    The goal and celebration is to increase the yield and preserve the native properties of globin.
    The goal is achieved by adjusting the pH of the initial solution to 2.5-4.5, then treating it with ethanol at a concentration of 40% by volume and sodium chloride, precipitated hemin is removed, and globin is precipitated from the solution. and separated by centrifugation.
    The method is carried out as follows. 20
    First, a part of the iron-containing component is separated, which exists in the form of aggregation in concentrated solutions or spores on the progestin component: JIO oi.iijiciiiii ocvHioi
    trifugation. Sodium chloride is added, and then globin is deposited in the resulting upper layer by adding a substance to this layer that can increase ionic strength or dehydration while the iron-containing component remains in the liquid phase in a liquid nln dispersed state.
    The medium created by the high content of solvent, for example ethanol, and low water content, with a certain ionic strength and low pH value, is favored by the fact that hemin and its derivatives, as well as its decomposition products, are retained in the liquid states so that globin can be separated by centrifugation.
    权利要求:
    Claims (2)
    [1]
    Example 1. To a hemoglobin solution with a dry matter content of 15% and ethanol of 33% at -8 ° C is added dropwise while moving and cooling a mixture of 1.1 ml of 1 M hydrochloric acid and 5 ml of water. After this addition, the temperature is -7 ° C, pH 2.5. 80 ml of 94% ethanol is added dropwise to this mixture, cooling it to -10 ° C. Globin precipitates upon adding 0.3 ml of ammonium sulfate to the mixture in the form of a 20% aqueous solution and centrifuged for 10 minutes at 8000 d. The surface liquid layer has a bright brown color and the precipitated gray color. This precipitate is washed in 85% ethanol at -10 ° C and pH 3.1, then centrifuged, resulting in a gain of 2.4 g of mass with a dry matter content of 23%. Example
    [2]
    2. In a solution of 36 ml of 94% ethanol, 6 ml of water and 0.55 ml of 1 M hydrochloric acid at -12 ° C are added dropwise while stirring and cooling 2.5 g of hemoglobin solution, the temperature remains -12 ° C, pH 3.0. Then 45 ml of 94% ethanol is added to the mixture and the liquid is centrifuged for 1 min at 8000 d, a thick mass of 0.1 g is obtained. Globin precipitates by adding 0.4 ml of a 20% ammonium sulfate solution. The pH was adjusted to 3.0. By centrifuging in the same manner as in Example 1, 45 g of a thick mass with a dry matter content of 36% was obtained. Further precipitation from the surface liquid layer is carried out by adding 0.4 ml of ammonium sulfate, 9.40 g of mol are obtained with a dry matter content of 42%. The resulting thick mass of light gray, the surface layer of bright brown, does not form a precipitate upon the introduction of trichloroxy acid. Example 3. To a solution of 20 ml of 94% ethanol and 1.1 ml of 1 M hydrochloric acid at -15 ° C, 5 g of a hemoglobin solution, pH 2.9, are added dropwise with simultaneous stirring and cooling. The mixture was centrifuged at 27,000 g for 20 minutes, a thick black mass of 0.9 g was formed with a dry matter content of 8.3%. With stirring and cooling, 50 ml of 94% ethanol is added to the superficial brown layer; upon addition of 10 ml of water and 1.2 ml of 20% ammonium sulfate, a precipitate is formed. The precipitate was separated by centrifugation at 8000-9 for 10 minutes and the resulting thick mass was suspended in 94% ethanol at -15 ° C and again separated by centrifugation under the same conditions. 1.6 g of a light gray mass are obtained with a dry matter content of 37.6%. Example 4. To a solution of 40 ml of 94% ethanol and 1.1 ml of 1M hydrochloric acid at -15 ° C, 5 g of hemoglobin solution, pH 2.0, are added dropwise with stirring and cooling. To this mixture is added dropwise 5 ml of 20% sodium chloride and centrifugal liquid at 27,000 g for 20 minutes, to obtain 1.4 g of a black solid with a dry matter content of 11%. 12 ml of water and 1 ml of a 20% ammonium sulphate solution are added to this surface layer, and the liquid is then centrifuged at 8000 g for 10 minutes. Get a gray mass in the amount of 3.8 g with a dry matter content of 14.6%. Example5. K5ml of the hemoglobin solution at -8 ° C is added dropwise while stirring and cooling a mixture of 11 ml of water, 4 ml of ethanol and 1 ml of 1 M hydrochloric acid, the temperature of the mixture is -6 ° C. After the addition, the temperature is -4 ° C, pH 2.9. Adding 7 ml of a 0.1 M solution of sodium hydroxide pH 3.9. Adding 0.2 g of ammonium sulfate precipitates as a brown mass. The precipitate is separated by centrifugation at 300 d. The liquid mass is suspended several times in small amounts of 94% ethanol at pH 3.0 and a temperature of -12 ° C and separated by centrifugation. Get 3.05 g of gray mass with a dry matter content of 18.8%. Example 6. From 5 g of hemoglobin solution, hemoglobin is precipitated by adding 10 ml of 94% ethanol to this solution with stirring and precipitating} shi at -6 ° C. The precipitate formed is separated by centrifugation for 5 minutes at 2000 g, 6 g of a thick red mass are obtained. A black precipitate is formed at the bottom of the centrifuge. The red mass is added to -70 ml of a solution of 94% ethanol and 1.1 ml of 1 M hydrochloric acid at -12 ° C with stirring and cooling. After adding the pH of the solution is 2.9, the temperature is -12 ° C. The mixture is centrifuged for 10 minutes at 27,000 g, a small amount of black mass is formed. A non-precipitated layer of liquid is added to 2 ml of a 40% ammonium sulfate solution and 10. ml of water. The temperature of this mixture is -8 ° C, pH 3.4. The deposited precipitate is separated by centrifugation for 10 minutes at 800 d. 1.5 g of a thick gray matter with a dry matter content of 93.3% are obtained. PRI and MER 7. To 20 ml of a 94% ethanol solution and 1.1 ml of 1 M hydrochloric acid at -12 ° C, 5 g of hemoglobin solution is added dropwise with stirring and cooling. After adding pH 2.9, the temperature is -12 ° C. The resulting mixture of sterno-black measles is centrifuged for 10 min at 27000 g, 2.2 g of the flesh is obtained with an ear content of 2.7%; emerging on top of Hocri., the light brown liquid acquires gel gel: 1 consistency. To the non-precipitated liquid layer are added dropwise with stirring and cooling 70 ml of cold ethanol, and then 0.8 ml of 40% ammonium sulphate solution and 8 ml of water. Thereafter, the temperature is -6 ° C. The precipitated precipitate is separated by centrifugation for 10 min at 8000 d, 1.7 g of a white solid with a dry matter content of 36% is obtained. The proposed method provides a high yield of the product while maintaining the native properties of globin. 6. The claims of the method of separating hemo iron from globin, including the step-by-step processing of the solution containing hemoglobin with salts and organic solvents, characterized in that, in order to increase the yield and preserve the native properties globally, the pH of the initial solution is adjusted to 2 , 5-4.5, then it is treated with ethanol in a concentration of 40–91% by volume and sodium chloride, precipitated hemin is removed, and globin is precipitated by the solution and separated by centrifugation .® Information sources taken in Attention during examination 1. Bresler S. E. Vve in molecular biology. 1973, p. 130-131.
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    同族专利:
    公开号 | 公开日
    BR7503722A|1976-06-29|
    ZA753806B|1977-01-26|
    FR2274230B1|1981-03-27|
    RO63791A|1978-12-15|
    AR209770A1|1977-05-31|
    FI59195B|1981-03-31|
    FR2274230A1|1976-01-09|
    SE7407882L|1975-12-15|
    AU502954B2|1979-08-16|
    ATA456575A|1978-03-15|
    SE389596B|1976-11-15|
    AU8166875A|1976-12-02|
    HU174189B|1979-11-28|
    CA1055489A|1979-05-29|
    AT346672B|1978-11-27|
    DK150173B|1986-12-29|
    YU147475A|1982-05-31|
    DE2526596C2|1984-11-29|
    DE2526596A1|1976-01-02|
    IE49166B1|1985-08-21|
    ES438483A1|1977-02-01|
    FI751643A|1975-12-15|
    FI59195C|1981-07-10|
    DK266675A|1975-12-15|
    PL94776B1|1977-08-31|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    DE178902C|1905-10-16|
    SE348942B|1970-06-02|1972-09-18|Statens Bakteriologiska Labor|CA1126653A|1978-12-22|1982-06-29|Jan H. Luijerink|Process of preparing blood cell protein and hemefrom hemoglobin|
    SE440596B|1980-04-03|1985-08-12|Paul Goran Sigvard Lindroos|PROCEDURE FOR PREPARING A HOME CONCENTRATE FROM A MIXTURE OF HOME AND BLOOD SUBSTANCE RECOVERY BY DIVISION OF HEMOGLOBIN|
    FR2535173A1|1982-11-03|1984-05-04|Protein Sa|Products obtained from the blood of abattoir animals and method for obtaining them.|
    JPS59128337A|1983-01-11|1984-07-24|Riyoushiyoku Kenkyukai|Recovery of blood globin and haem|
    FR2548671B1|1983-07-07|1986-05-02|Merieux Inst|PROCESS FOR THE PREPARATION OF A GLOBIN FROM HEMOGLOBIN AND A GLOBIN OBTAINED BY THIS PROCESS|
    DE3608091A1|1986-03-12|1987-09-17|Basf Ag|METHOD FOR INSULATING AND CLEANING HAEMIN|
    DE68918083T2|1988-11-18|1995-01-05|Tosoh Corp|METHOD FOR PURIFYING BLOOD PLASMA.|
    ES2181046T3|1996-12-20|2003-02-16|Fraunhofer Ges Forschung|PROCEDURE FOR THE OBTAINING OF FEMALE FROM BLOOD OF KILLING.|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    SE7407882-5A|SE389596B|1974-06-14|1974-06-14|SUGGESTED TO SEPARATE GLOBIN IRON COMPOUNDS FROM A VETICAL CONTAINING BLOOD SUBSTANCES|
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