Heparin extraction method

Abstract

Provided are: a heparin extraction method enabling extraction of heparin from an organ while maintaining an edible state of the organ; an edible organ with a low heparin content and a method for producing the same; an organ treatment method; and an organ treatment system.　The present invention makes it possible to extract heparin from an organ while maintaining an edible state of the organ by subjecting the organ to an alkali treatment at a low temperature. The extracted heparin can be utilized as a medicine or the like, and the organ after the extraction can be utilized as food.

Description

Heparin extraction method

[0001] This invention relates to a heparin extraction method, edible organs with low heparin content and methods for producing the same, organ processing methods and organ processing systems.

[0002] Heparin is a substance used in pharmaceuticals and other products, such as those for preventing blood coagulation. Heparin is known to be contained in organs such as the small intestine and liver of animals such as pigs and cattle, and the current mainstream method for producing heparin involves treating animal organs such as pigs and cattle with enzymes or alkalis (Patent Documents 1-3, Non-Patent Document 1).

[0003] Comparative Biochemistry and Physiology Part C 136 (2003) 357-365

[0004] JP-A-55-164201 JP-A-55-164202 Patent No. 4455680

[0005] Conventional heparin production methods involve dissolving most of the organ used as raw material while extracting heparin. As a result, the organs after heparin extraction cannot be used as food and are either used as animal feed or discarded. This invention aims to enable both heparin extraction from organs and the maintenance of the organs in an edible state, in order to effectively utilize the organs after heparin extraction as food.

[0006] As a result of diligent research, the inventors discovered that the above problems can be solved by alkaline treatment of organs at low temperatures, and have completed the following invention.

[0007] This application provides the following inventions: (1) An edible organ characterized by having a heparin content of 10 mg / kg or less per organ. (2) A heparin extraction method characterized by comprising the step of extracting heparin from an organ obtained from a non-human animal by alkaline treatment at a temperature of 10°C or lower. (3) A method for producing an edible organ, characterized by comprising the step of producing an edible organ by alkaline treatment at a temperature of 10°C or lower from an organ obtained from a non-human animal. (4) A method for processing an organ, characterized by comprising the steps of: providing heparin, which includes extracting heparin from an organ obtained from a non-human animal by alkaline treatment at a temperature of 10°C or lower; and providing an edible organ, which includes obtaining the organ after the alkaline treatment as an edible organ. (5) The method according to (3) or (4), characterized in that the heparin content of the edible organ obtained by the above method is 10 mg / kg or less per organ. (6) An organ processing system comprising: a heparin providing means for extracting and providing heparin from organs obtained from non-human animals by alkaline treatment at a temperature of 10°C or lower; and an edible organ providing means for obtaining the organs after the alkaline treatment as edible organs. (7) Edible organs obtained by the method described in any one of (3) to (5) or by the system of (6).

[0008] This invention makes it possible to extract heparin from organs and maintain the organs in an edible state. The extracted heparin can be used as a medicine, and the organs after extraction can be used as food.

[0009] Figure 1 is a photograph showing the appearance of the small intestine before and after heparin extraction in Example 4-2. Figure 2 shows the shear force (N) of the small intestine before and after heparin extraction in Example 4-3. Figure 3 is a photograph showing the appearance of the small intestine after extraction with each extract in a 90°C bath for 30 minutes in Example 5-1. Figure 4 is a photograph showing the appearance of the small intestine before extraction and after extraction overnight at 5°C with 0.13 mol / L, 0.23 mol / L, and 0.3 mol / L NaOH aqueous solutions in Example 5-2. Figure 5 shows the weight of the small intestine before extraction and after extraction at 16 hours, 24 hours, 48 ​​hours, and 72 hours in Example 5-3-4. Figure 6 shows the weight change of the small intestine after each extraction time in Example 5-3-4, with the weight before extraction set to 100%. Figure 7 is a photograph showing the appearance of the small intestine before extraction and after extraction at 16 hours, 24 hours, 48 ​​hours, and 72 hours in Example 5-3-4. Figure 8 shows the appearance of the small intestine after extraction at 16 hours, 24 hours, 48 ​​hours, and 72 hours in Example 5-3-4. Figure 9 shows the weight of the small intestine before extraction and after extraction at 16 hours, 24 hours, 48 ​​hours, and 72 hours in Example 5-3-5, along with the results for Example 5-3-4. Figure 10 shows the appearance of the small intestine after extraction at 16 hours, 24 hours, 48 ​​hours, and 72 hours in Example 5-3-5, along with the results for Example 5-3-4. Figure 11 shows the appearance of the small intestine after extraction using the extract at each temperature in Example 5-3-4.

[0010] As mentioned above, heparin is mainly produced by treating organs such as the small intestine and liver of animals such as pigs and cattle with enzymes or alkalis (Patent Documents 1-3, Non-Patent Document 1). However, when these conventional methods are used, the organs are severely damaged by enzyme treatment or high-temperature alkali treatment, resulting in dissolution, fragmentation, discoloration, liquefaction, and solification, making them unsuitable for consumption. In many cases, the organs remaining after heparin extraction cannot be effectively utilized and must be discarded. Furthermore, even if they are used, their use is limited to low-value-added applications such as animal feed.

[0011] However, according to the present invention, it is possible to extract heparin from organs and maintain the organs in an edible state after heparin extraction. The heparin extracted from the organs can be used as a pharmaceutical, etc., while the organs after heparin recovery can be used as a high-value-added food. In this specification, "pharmaceutical, etc." refers to all heparin-containing compositions such as anticoagulants and other pharmaceuticals, and quasi-drugs.

[0012] This invention makes it possible to make effective use of raw materials. Organs from pigs, cows, etc. (for example, the digestive tract such as the small intestine), which are the raw material for heparin, are widely used as ingredients in dishes such as offal stew, offal hot pot, and grilled offal. The edible organs after heparin extraction according to this invention (edible organs with a low heparin content) can maintain their shape from before extraction, making it possible to distribute them to the market without compromising their value as food ingredients. Furthermore, ordinary organs from pigs, cows, etc. are highly elastic and difficult to chew, resulting in a hard texture, making them difficult to use for consumption without pre-treatment such as boiling them several times in hot water. However, the edible organs after heparin extraction according to this invention change to an appropriate elasticity through the heparin extraction process, resulting in a texture suitable for consumption. In addition, there is no need to discard the organs after heparin recovery, which is advantageous in terms of the effort, time, and cost required for disposal.

[0013] One aspect of the present invention relates to a heparin extraction method characterized by including the step of extracting heparin from an organ obtained from an animal by alkaline treatment at a low temperature; a heparin production method characterized by including the step of extracting heparin from an organ obtained from an animal by alkaline treatment at a low temperature; a method for producing an edible organ characterized by including the step of producing an edible organ by alkaline treatment at a low temperature from an organ obtained from an animal; and an organ processing method characterized by including the step of providing heparin, which includes extracting heparin from an organ obtained from an animal by alkaline treatment at a low temperature, and the step of providing an edible organ, which includes obtaining the organ after the alkaline treatment as an edible organ (in this specification, these methods are collectively referred to as the methods of the present invention).

[0014] Furthermore, one aspect of the present invention relates to an organ processing system (in this specification, this method is referred to as the system of the present invention) characterized by comprising a heparin providing means for extracting and providing heparin from organs obtained from animals by alkaline treatment at low temperature, and an edible organ obtaining means for obtaining the organs after the alkaline treatment as edible organs.

[0015] The organ processing method or system of the present invention provides a completely new and unprecedented business that combines two aspects: a pharmaceutical-related business, on the one hand, providing heparin useful as a medicine, and a food-related business, on the other hand, providing heparin extraction residue, which would conventionally be discarded after heparin extraction, as a food product.

[0016] The animals used in the method or system of the present invention are not limited to animals that produce heparin. Examples include non-human animals such as pigs, cattle, horses, monkeys, and sheep, and fish such as sharks. As an example, pigs or cattle can be used because they allow for the extraction of heparin at a level usable as a pharmaceutical and are frequently used as food.

[0017] The organ is not limited to any organ that contains heparin. For example, heparin is present in the intestinal mucosa of the small and large intestines, the lungs, liver, and spleen, and is stored in mast cells in muscles, etc., so organs such as the small and large intestines, lungs, liver, and spleen, as well as muscles, etc., can be used. As an example, the small intestine can be used because it has a high heparin content and is used as food. The organ may be pre-processed by washing, boiling, cutting, etc., and may be used in a raw state or thawed frozen state. However, the pre-processing of the small intestine for producing natural casing (processing to remove specific tissues such as the muscular layer and leave only the submucosa) is not included in the pre-processing of the small intestine for producing natural casing. Therefore, the pre-processed small intestine that becomes the organ of the present invention represents multiple layers of small intestinal mesoplastic tissue (mucosa, submucosa, muscular layer, and serosal layer) including at least the mucosa. It is preferable to use the organ in a low temperature state suitable for low-temperature alkaline treatment before heparin extraction. The temperature can be any temperature that is equivalent to the temperature of the alkaline treatment or the temperature of the extract used in the alkaline treatment, for example, 0°C to 25°C, 0°C to 20°C, 0°C to 19°C, 2°C to 18°C, 3°C to 17°C, 4°C to 16°C, 0°C to 15°C, 5°C to 15°C, 6°C to 11°C, 7°C to 10°C, 7°C to 11°C, 9°C to 10°C, 7°C to 9°C, 7°C to 8°C, 8°C to 10°C, etc. From the viewpoint of suppressing organ damage or discoloration, or increasing the recovery rate of heparin, examples include 25°C or below, less than 25°C, 20°C or below, less than 20°C, 15°C or below, less than 15°C, 14°C or below, 13°C or below, 12°C or below, 11°C or below, 10°C or below, 9°C or below, 8°C or below, etc. On the other hand, from the perspective of enabling heparin extraction, suitable temperatures include 0°C or higher, 1°C or higher, 2°C or higher, 3°C or higher, 4°C or higher, 5°C or higher, 6°C or higher, 7°C or higher, etc.

[0018] In heparin extraction, the solution used for alkaline treatment is not limited as long as it is a solution that can extract heparin while maintaining the organ in an edible state. Examples include any alkaline solution containing one or more of the following: NaOH, KOH, NH3・H2O, Na2CO3, NaHCO3, Ca(OH)2, etc. The solvent is also not limited as long as it is a solution that can extract heparin while maintaining the organ in an edible state. Any solvent such as water can be used. For example, an aqueous NaOH solution can be used because it facilitates heparin extraction and causes little damage or discoloration to the organ.

[0019] The concentration of the solution used for alkaline treatment can be adjusted as appropriate depending on the type of alkaline solution used, its alkalinity, temperature, etc. For example, 0.001 mol / L to 100.0 mol / L, 0.001 mol / L to 10.0 mol / L, 0.005 mol / L to 5.0 mol / L, 0.01 mol / L to 0.1 mol / L, 0.05 mol / L to 0.3 mol / L, 0.05 mol / L to 0.25 mol / L, 0.05 mol / L to 0.2 mol / L, 0.1 mol / L to 0.3 mol / L. Examples include 0.1 mol / L, 0.1 mol / L to 0.25 mol / L, 0.1 mol / L to 0.2 mol / L, approximately 0.05 mol / L, approximately 0.09 mol / L, approximately 0.1 mol / L, approximately 0.2 mol / L, approximately 0.3 mol / L, approximately 0.5 mol / L, approximately 1.0 mol / L, approximately 2.0 mol / L, approximately 3.0 mol / L, approximately 5.0 mol / L, approximately 10.0 mol / L, etc.

[0020] For example, in the case of NaOH aqueous solution, the concentration of NaOH is 0.01 mol / L to 5.0 mol / L, 0.02 mol / L to 4.0 mol / L, 0.03 mol / L to 3.0 mol / L, and 0.04 mol / L to 2.0 mol / L. , 0.05 mol / L to 1.0 mol / L, 0.05 mol / L to 0.3 mol / L, 0.05 mol / L to 0.25 mol / L, 0.05 mol / L to 0.2 mol / L, 0.1 mol / L to 0.3 mol / L, 0.1 m ol / L ~ 0.25 mol / L, 0.1 mol / L ~ 0.2 mol / L, 0.06 mol / L ~ 0.3 mol / L, 0.07 mol / L ~ 0.25 mol / L, 0.08 mol / L ~ 0.2 mol / L, 0.09 mol / L ~0.2 mol / L, about 0.05 mol / L, about 0.07 mol / L, about 0.09 mol / L, about 0.1 mol / L, about 0.2 mol / L, about 0.3 mol / L, about 0.4 mol / L, about 0.5 mol / L, etc. can be adopted. For example, from the viewpoint of increasing the heparin recovery rate while suppressing organ damage and discoloration, ranges such as 0.05 mol / L or more and less than 0.3 mol / L, 0.05 mol / L or more and 0.28 mol / L or less, 0.05 mol / L or more and 0.25 mol / L or less, 0.05 mol / L or more and 0.2 mol / L or less, 0.1 mol / L or more and less than 0.3 mol / L, and 0.1 mol / L or more and 0.2 mol / L or less can be preferably adopted. From the viewpoint of suppressing organ damage, upper limits can be 0.3 mol / L or less, less than 0.3 mol / L, less than 0.28 mol / L or less, less than 0.28 mol / L, less than 0.25 mol / L or less, less than 0.25 mol / L, less than 0.2 mol / L, less than 0.2 mol / L, and 0.1 mol / L or less. On the other hand, from the perspective of increasing the heparin recovery rate, the lower limit can be defined as greater than 0.01 mol / L, 0.01 mol / L or more, 0.02 mol / L or more, 0.03 mol / L or more, 0.04 mol / L or more, 0.05 mol / L or more, greater than 0.05 mol / L, 0.06 mol / L or more, 0.07 mol / L or more, 0.08 mol / L or more, 0.09 mol / L or more, etc.

[0021] The pH of the solution used for alkaline treatment is not limited as long as it is a pH that can extract heparin while keeping the organs in an edible state, and can be adjusted as appropriate depending on the type of alkaline solution used, temperature, treatment time, etc. Examples include 8-14, 9-14, 10-14, 11-14, 12-14, 10-13.8, 10-13.5, 10-13.3, 11-13.6, 11.3-13.8, 11.5-13.5, 11.7-14, 12-14, 12-13.5, 12.5-13.5, 12.7-13.5, 13-13.5, etc. From the perspective of minimizing damage to organs, examples include 13.8 or less, 13.7 or less, 13.6 or less, 13.5 or less, 13.4 or less, 13.3 or less, etc. On the other hand, from the perspective of increasing the heparin recovery rate, values ​​such as 10 or more, 11 or more, 11.5 or more, 12 or more, 12.1 or more, 12.2 or more, 12.3 or more, 12.4 or more, 12.5 or more, 12.7 or more, and 13 or more are mentioned.

[0022] It is preferable to use an alkaline solution that is at a low temperature suitable for low-temperature alkaline treatment before heparin extraction. The temperature should be approximately the same as the temperature of the alkaline treatment or the temperature of the organ used for the alkaline treatment, for example, 0°C to 25°C, 0°C to 20°C, 0°C to 19°C, 2°C to 18°C, 3°C to 17°C, 4°C to 16°C, 0°C to 15°C, 5°C to 15°C, 6°C to 11°C, 7°C to 10°C, 7°C to 11°C, 9°C to 10°C, 7°C to 9°C, 7°C to 8°C, 8°C to 10°C, etc. From the viewpoint of suppressing organ damage or discoloration, or increasing the heparin recovery rate, suitable temperatures include 25°C or below, less than 25°C, 20°C or below, less than 20°C, 15°C or below, 14°C or below, 13°C or below, 12°C or below, 11°C or below, 10°C or below, 9°C or below, 8°C or below, 7°C or below, etc. On the other hand, from the perspective of enabling heparin extraction, suitable temperatures include 0°C or higher, 1°C or higher, 2°C or higher, 3°C or higher, 4°C or higher, 5°C or higher, 6°C or higher, 7°C or higher, etc.

[0023] Alkaline treatment is performed at low temperatures. In this specification, low temperature refers to a temperature at which heparin can be extracted while maintaining the organ in an edible state, and can be appropriately adjusted according to the type, strength, concentration, temperature, pH of the solution, treatment time, desired edible use and texture of the organ after treatment. For example, temperatures such as 0°C to 25°C, 0°C to 20°C, 0°C to 19°C, 1°C to 18°C, 1°C to 17°C, 2°C to 16°C, 2°C to 15°C, 3°C to 10°C, 3°C to 9°C, 4°C to 10°C, 4°C to 9°C, 5°C to 9°C, 5°C to 10°C, 6°C to 10°C, 7°C to 10°C, 7°C to 11°C, 9°C to 10°C, 7°C to 9°C, 7°C to 8°C, 8°C to 10°C, etc., can be arbitrarily adopted. From the perspective of suppressing organ damage or increasing the recovery rate of heparin, suitable temperatures include 25°C or below, less than 25°C, 20°C or below, less than 20°C, 15°C or below, 14°C or below, 13°C or below, 12°C or below, 11°C or below, 10°C or below, 9°C or below, and 8°C or below. On the other hand, from the perspective of enabling heparin extraction, suitable temperatures include 0°C or above, 1°C or above, 2°C or above, 3°C or above, 4°C or above, and 5°C or above.

[0024] The extraction time by alkaline treatment can be appropriately adjusted according to the type, strength, concentration, and temperature of the alkaline solution, as well as the desired edible use and texture of the organ after treatment. For example, any time can be adopted, such as 1 to 70 hours, 2 to 48 hours, 2 to 36 hours, 4 to 36 hours, 6 to 36 hours, 16 to 36 hours, 2 to 24 hours, 4 to 24 hours, 6 to 24 hours, 2 to 20 hours, 4 to 20 hours, 6 to 20 hours, 16 to 24 hours, 2 to 16 hours, 4 to 16 hours, 6 to 16 hours, 2 to 12 hours, 4 to 12 hours, 6 to 12 hours, 2 to 10 hours, 2 to 8 hours, 2 to 6 hours, 2 to 4 hours, 4 to 11 hours, 6 to 10 hours, 6 to 8 hours, 4 to 8 hours, etc. For example, from the standpoint of suppressing organ damage, maintaining an edible state, or preserving suitability for processing, examples include less than 72 hours, 48 ​​hours or less, less than 48 hours, 24 hours or less, 20 hours or less, 16 hours or less, 12 hours or less, and 10 hours or less. On the other hand, from the standpoint of increasing the heparin recovery rate, examples include 2 hours or more, 3 hours or more, 4 hours or more, 5 hours or more, and 6 hours or more.

[0025] As described above, the alkaline treatment is not limited as long as it is possible to extract heparin while maintaining the organs in an edible state. For example, an aqueous NaOH solution of less than 0.3 mol / L, such as about 0.05 mol / L, about 0.1 mol / L, or about 0.2 mol / L, can be used to treat the organs at a temperature of about 10°C or lower, such as 3 to 10°C, for a period of 2 to 36 hours, 6 to 24 hours, 6 to 20 hours, 6 to 16 hours, or 6 to 12 hours, for a period of 2 hours or more but less than 48 hours.

[0026] The method or system of the present invention may include a step or means of pre-treating the organ, such as washing, boiling, or cutting, before alkaline treatment. It may also include a step or means of recovering, measuring, and / or purifying heparin after heparin extraction. For example, heparin can be recovered and its amount measured by the following method: The heparin extract can be decomposed using an enzyme such as alcalase, filtered by suction, then an ion exchanger is added and sieved, washed, and eluted to recover the heparin. The amount of heparin thus obtained can be measured, for example, by liquid chromatography. For the purification of heparin, any method can be employed, such as precipitating crude heparin by adding ethanol or the like, removing impurities using an ion exchanger, eluting from the exchanger at a specific ion concentration or pH, filtration, ultrafiltration, dialysis, or sterilization. However, the pre-treatment of the organ, measurement of heparin amount, recovery, and purification are not limited to these methods, and any method can be used.

[0027] The method or system of the present invention may include steps or means for extracting heparin from an organ. For example, steps or means may be employed to appropriately monitor and control the temperature, pH, etc., of the alkaline treatment to an optimal state, or to check the heparin extraction state during treatment, such as the heparin concentration in the alkaline solution or the hardness of the organ during treatment, and to perform treatment according to that state. Such steps or means may be performed by training a neural network with data.

[0028] The method or system of the present invention may use data relating to reference values ​​such as the heparin-containing site or region in an organ, the heparin content contained in the organ, and the stiffness of the organ. By comparing such data with the subject being processed, it becomes possible to obtain heparin and / or organs with desired requirements such as desired quantity, quality, or time. Such data may be obtained by training a neural network.

[0029] The method or system of the present invention may include steps or means for neutralizing the organ after alkali treatment, steps or means for washing the organ after alkali treatment or neutralization, steps or means for processing it to make it suitable for food, etc. Neutralization is sufficient as long as it can neutralize the alkaline solution used for heparin extraction. For example, any acidic solution containing one or more of the following can be used: HCl, CH3COOH, C6H8O7, H2CO3, etc., and can be appropriately selected depending on the concentration of the alkaline solution used for heparin extraction, the type of alkali, the desired pH of the solution after neutralization, etc. For example, aqueous solutions of HCl with concentrations of 1.0 mol / L to 10.0 mol / L, 2.0 mol / L to 9.0 mol / L, 4.0 mol / L to 8.0 mol / L, approximately 6.0 mol / L, etc. can be used. The pH of the solution after neutralization is not limited and can be appropriately adjusted depending on the intended use of the organ as food, etc. For example, values ​​such as 2-10, 3-9, 4-8, 5-8, 2-7, 4-7, 7-9, 6-7, 7-8, 6-8, 6.5-7.5, 6.5-7, 7-7.5, approximately 6.6, approximately 6.7, approximately 6.8, approximately 6.9, approximately 7.0, approximately 7.1, approximately 7.2, approximately 7.3, approximately 7.4, etc., can be arbitrarily adopted. For washing, any washing solution and washing method such as water or salt water can be used. Processing to make it suitable for consumption includes, for example, the addition of any additives such as seasonings, fragrances, flavorings, colorings, preservatives, thickeners, pH adjusters, etc., to make it suitable for use by secondary processors or consumers, cutting, boiling, blanching, simmering, steaming, baking, roasting, smoking, freezing, refrigeration, retort processing, pouch packaging, canning, bottling, etc.

[0030] For example, steps or means may be employed to appropriately monitor the type, pH, concentration, and temperature of the alkaline solution and manage the type, pH, concentration, and temperature of the acidic solution to neutralize it to the optimal state; to check the turbidity of the solution during treatment, the degree of contamination of the organ after alkaline treatment, neutralization, or washing, and manage the type of washing solution, washing method, and time according to the condition; to check the hardness, elasticity, hardness, gum-like properties, and chewiness of the organ after alkaline treatment, neutralization, or washing, and manage the processing such as boiling, simmering, or simmering, and the number of times such processing is performed according to the condition; and to remove specific tissues such as the muscle layer.

[0031] In steps or means of neutralizing organs after alkaline treatment, washing organs after alkaline treatment or neutralization, processing organs to make them suitable for consumption, etc., data on standard values ​​such as the type of alkaline or acidic solution, pH, concentration, temperature, turbidity of the solution, degree of organ contamination, type of washing solution, washing method, time, elasticity, hardness, gumminess, chewiness of the organs, and type and number of processing steps may be used. By comparing such data with the target during processing, it becomes possible to obtain edible organs with desired requirements such as desired quantity and quality or time. Such data may be obtained by training a neural network.

[0032] One aspect of the present invention is an edible organ characterized by a low heparin content, or an edible organ characterized by being obtained by the method or system of the present invention (in this specification, these edible organs are collectively referred to as the edible organs of the present invention).

[0033] A low heparin content means that it is lower than that of an organ in which heparin extraction is not performed using the method of the present invention. For example, compared to the heparin content per organ in which heparin extraction is not performed, the heparin content in the organ according to the present invention may be 1% to 95%, 2% to 90%, 3% to 80%, 4% to 70%, 5% to 60%, 6% to 50%, 7% to 40%, 8% to 30%, 9% to 25%, 10% to 20%, or as an upper limit, 90% or less, 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, etc. While there is no lower limit to the heparin content, the present invention only applies alkali treatment to an extent that maintains the organ in an edible state. Therefore, the heparin content may be slightly higher compared to cases where strong alkali treatment is performed, such as prolonged or high-temperature treatment that renders the organ unsuitable for consumption. From this perspective, for example, the heparin content in the organ of the present invention may be 0.5% or more, 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, etc., compared to the heparin content per organ without heparin extraction. For example, when using pig small intestines, the heparin content per small intestine is usually around 110 mg / kg to 180 mg / kg, and around 150 mg / kg to 200 mg / kg after fat has been removed. However, the heparin content in the small intestine of the present invention may be, for example, 1 mg / kg to 100 mg / kg, 2 mg / kg to 90 mg / kg, 3 mg / kg to 80 mg / kg, 4 mg / kg to 70 mg / kg, 5 mg / kg to 60 mg / kg, 6 mg / kg to 50 mg / kg, 7 mg / kg to 40 mg / kg, 8 mg / kg to 30 mg / kg, 9 mg / kg to 25 mg / kg, 10 mg / kg to 20 mg / kg, or as an upper limit, 100 mg / kg or less, 50 mg / kg or less, 20 mg / kg or less, 10 mg / kg or less, 9 mg / kg or less, 8 mg / kg or less, etc. While the lower limit is not limited, for the same reasons as described above, when using pig small intestines, the heparin content in the small intestine after heparin extraction according to the present invention may be, for example, 1 mg / kg or more, 2 mg / kg or more, 3 mg / kg or more, 4 mg / kg or more, 5 mg / kg or more per small intestine.The heparin content in an organ can be determined by decomposing the organ using an enzyme such as alcalase, eluting and recovering the heparin, and then measuring the amount of recovered heparin, for example, by liquid chromatography.

[0034] The edible organs of the present invention are advantageous in that they maintain an edible state while having a crisper texture compared to organs that have not undergone heparin extraction. For example, when preparing offal such as small intestines, it is often necessary to boil them two to three times during the preparation process to make them easier to chew. However, by using the edible organs of the present invention, it becomes possible to reduce or eliminate boiling and other processes that improve crispness, ease of chewing, and softness. By using edible organs that have a crisp texture while maintaining an edible state, they can be used not only as animal feed as in the conventional technology, but also as highly palatable ingredients that can be used in dishes for human consumption. Furthermore, in this specification, edible organs are a different concept from natural casings used as packaging materials for filling with meat fillings when manufacturing processed products such as sausages. Natural casings utilize hollow organs such as animal intestines, and are produced by scraping off and washing away the fat, muscle layer, mucous layer, etc. attached to the organ, thereby utilizing the tough submucosal layer (for example, "Science of Meat and Meat Products," March 31, 1992, 1st edition, Gakusosha, p. 171, and "Meat Processing Handbook," November 20, 1980, Tsuneo Kamata, p. 358, etc.). On the other hand, unlike such natural casings, the edible organs of the present invention retain their tissue structure (for example, in the case of the small intestine, the mucous layer, submucosa, muscle layer, and serosal layer, etc.) and remain edible unless pre-processing, steps, or means are performed to remove specific tissues. Therefore, the edible organs in the present invention do not include casings.

[0035] Maintaining an edible state means that the organ is not severely damaged, such as dissolution, fragmentation, discoloration, liquefaction, solification, or expansion, and that the shape and tissue structure of the organ remain intact, and that it retains its texture. For example, even if the organ has suffered damage such as surface dissolution, if its shape, tissue structure, and texture are still present, it can be said that it is still edible. For example, in the case of the small intestine, even if there is damage to the surface, if the mucosa, submucosa, muscular layer, and serosa remain intact, the shape and tissue structure can be said to remain intact. Shape and tissue structure can be measured by visual inspection or image analysis, and texture can be measured by arbitrary indicators such as elasticity, hardness, gum-likeness, and chewiness. On the other hand, if the texture is too hard, it is difficult to chew, so it is also important that it has a certain degree of softness, ease of chewing, or crispness. Softness, ease of chewing, or crispness can be measured, for example, by sensory evaluation or shear force. The edible organs of the present invention have a lower shear force compared to organs that have not undergone heparin extraction. In one embodiment, the edible organs of the present invention have lower shear force values ​​of 5N to 35N, 8N to 25N, 10N to 20N, 10N to 30N, approximately 10N, approximately 20N, and approximately 30N compared to organs that have not undergone heparin extraction. Examples of appropriate shear force values ​​for an edible state include 5N to 60N, 6N to 55N, 7N to 50N, 8N to 40N, 10N to 40N, 10N to 30N, 30N to 40N, 5N to 15N, and 20N to 40N. However, the shear force value varies depending on the intended use of the desired organ as food and is not limited to the above ranges. The shear force value can be measured using any device, such as a texture analyzer or universal tester. Sensory evaluation can be performed using any index, for example, by trained panelists. In the case of the small intestine, it is preferable that some fat remains from the viewpoint of palatability. The fat content can be measured using any indicator, such as visual inspection, ether extraction, or acid hydrolysis. Maintaining suitability for processing means that the organ remains edible even after further processing such as freezing, thawing, or reboiling following heparin extraction.

[0036] One aspect of the present invention is heparin characterized by being obtained by the heparin extraction method of the present invention, the step of providing heparin in the method for producing an edible organ, or by the heparin providing means in an organ processing system (in the present specification, these heparins are collectively referred to as heparin obtained by the method or system of the present invention).

[0037] Although the recovery rate of heparin obtained by the method or system of the present invention is not limited, for example, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, etc. can be mentioned. Although a higher recovery rate of heparin is preferable from the viewpoint of heparin provision, the type, strength, concentration, temperature, type, treatment temperature, pH, etc. of the alkaline solution may be appropriately adjusted in consideration of the desired edible use of the processed organ.

[0038] In the present specification, "about" includes errors and variations within the range that those skilled in the art understand to be acceptable for achieving the effects of the present invention with respect to the described numerical values, concentrations, amounts, times, temperatures, shear force values, and other measured values. Usually, this range is within ±10% of the described value, more preferably within ±5%. However, if the appropriate variation range recognized by those skilled in the art differs depending on the measurement item and implementation conditions, "about" shall be interpreted within the corresponding range.

[0039] Next, the present invention will be described in more detail with reference to examples. Note that the present invention is not limited thereby.

[0040] Example 1: Heparin extraction was performed using pig small intestine after washing and boiling. 1 kg of small intestine was immersed in 6.6 times the amount of 0.2 N NaOH aqueous solution and subjected to alkali treatment at 10°C or lower overnight (about 16 hours) to extract heparin. The extract and the residual small intestine were separated. The extract was used in Example 2, and the residue was used in Example 3.

[0041] Example 2: Purification and Measurement of Heparin in the Extract To the extract obtained in Example 1, 2.4 LFG of alkaline protease (Novozyme) at 1% by weight of the extract was added, and enzymatic treatment was carried out at 50 °C overnight. Then, the enzyme was inactivated by boiling at 90 °C for 30 minutes, and purification was performed using an anion exchange resin (PA308, Mitsubishi Chemical Aqua Solution). The purified liquid was used to precipitate heparin with ethanol (99.5% by weight), and after drying, the obtained powder was obtained as heparin powder.

[0042] The yield of the obtained heparin was measured by the following HPLC analysis. The HPLC method described in the confirmation test and purity test (7) of Japanese Pharmacopoeia Heparin Sodium was carried out under the following conditions. A calibration curve was created using a heparin sodium standard product, and the amount of heparin was determined by quantifying the amount of heparin sodium. HPLC equipment: Shimadzu LC-2030 Column used: TSK-Gel DEAE-5PW (φ2 mm × 7.5 cm, particle size 10 μm) Column temperature: 35 °C Flow rate: 0.2 ml / min Detection wavelength: 202 nm Sample injection volume: 20 μl Mobile phase A: A liquid obtained by dissolving 0.4 g of sodium dihydrogen phosphate dihydrate in 1 L of water, adjusting the pH to 3.0 with phosphoric acid, and passing through a 0.5 μm PTFE filter was used as mobile phase A. Mobile phase B: A liquid obtained by dissolving 0.4 g of sodium dihydrogen phosphate dihydrate and 106.4 g of lithium perchlorate in 1 L of water, adjusting the pH to 3.0 with phosphoric acid, and passing through a 0.5 μm PTFE filter was used as mobile phase B. Calibration curve: A calibration curve was created by diluting a 1 mg / ml heparin sodium standard product 2, 4, 6, 8, and 16 times.

[0043] As a result, it was found that 120 mg / kg of heparin could be extracted per 1 kg of small intestine.

[0044] Example 3: Measurement of Heparin Content in the Residue of Small Intestine after Heparin Extraction The small intestine remaining as a residue after heparin extraction in Example 1 was immersed in a solution containing 2.4 LFG of alkaline protease (Novozyme) at 1% by weight at 50 °C for 21 hours to dissolve it, and the heparin content of the obtained solution was measured in the same manner as in Example 2.

[0045] As a result, it was confirmed that the residual small intestine after extraction contained 7.8 mg / kg of heparin per small intestine. Therefore, it was found that heparin was extracted with a recovery rate of 93.9% by the method of Example 1.

[0046] Example 4: Characterization and processing of residual small intestine after heparin extraction Example 4-1: Neutralization The residual small intestine after heparin extraction in Example 1 was neutralized by adding 6N HCl in an equimolar amount to the NaOH used for extraction. After solid-liquid separation, it was washed with water, frozen and stored at approximately -18°C, thawed, and then subjected to Examples 4-2 to 4-4 below.

[0047] Example 4-2: Shape Evaluation The appearance of the residual small intestine before heparin extraction in Example 1 and after neutralization in Example 4-1 was visually inspected. As a result, the small intestine after heparin extraction had an appearance, shape, and tissue structure that was almost the same as before heparin extraction, although the surface was slightly smoother (Figure 1).

[0048] Example 4-3: Evaluation of Shear Force Normally, in order to make offal easier to chew, it is necessary to boil it two to three times during the preparation of offal. Therefore, in order to investigate whether the boiling process can be shortened by using the small intestine residue after heparin extraction, the shear force of the small intestine after heparin extraction was evaluated using the following method.

[0049] The small intestine residues from Example 1 (before heparin extraction) and Example 4-1 (after neutralization) were boiled at 90°C for 20 minutes and allowed to return to room temperature. This process was repeated three times for the small intestine residues from Example 1 (before treatment) and once for the small intestine residues from Example 4-1 (after neutralization). After each process, the shear force was evaluated using an Instron® shear force measuring device (N=10).

[0050] The results are shown in Figure 2. As shown in Figure 2, the small intestine after heparin extraction had a 46.5% reduction in shear force compared to the small intestine before heparin extraction, even without boiling. Furthermore, it was confirmed that the residual small intestine after heparin extraction had a lower shear force than the small intestine before heparin extraction when boiled two or three times, and was considerably easier to chew, even with just one boiling.

[0051] Example 4-4: Sensory Evaluation The small intestine residue from Example 1 before heparin extraction and from Example 4-1 after neutralization was boiled at 90°C for 20 minutes (either once or without this step), and then grilled over medium heat for about 5 minutes. Six trained panelists performed a sensory evaluation using the small intestine before heparin extraction as the standard, with the following five-point scale: 1: Poor, 2: Somewhat poor, 3: Equivalent to the standard, 4: Somewhat good, 5: Good. The average of the comments and scores from the six panelists are shown in Table 2.

[0052] These results show that using organs from heparin extraction residue can result in organs with an improved texture that are softer and easier to chew. Furthermore, it was found that adopting this invention makes it possible to reduce or eliminate the boiling process required to make the organs easier to chew.

[0053] Example 5: Examination of Extraction Conditions Based on the results of Examples 1 to 4, it was found that the present invention makes it possible to provide heparin useful as a medicine, etc., while also providing heparin extract residue as a food product. Next, the extraction conditions were examined using the following method.

[0054] Example 5-1: Solvent Examination A raw, frozen pig small intestine was used. The extract contained 1% by weight of Na ２ CO ３ Aqueous solution, 0.1 mol / L carbonate buffer (Na ２ CO ３ and NaHCO ３ Alkaline solutions were used, consisting of equimolar aqueous solutions (pH 10), a 0.05 mol / L NaOH aqueous solution, and a 0.01 mol / L NaOH aqueous solution.

[0055] 100 g of small intestine was immersed in three times the volume of each of the above extracts and kept in a 90°C bath for 30 minutes. After cooling with water, the extracts and residual small intestine were separated. The extracts were measured using the same method as in Examples 2 and 3, and the amount of heparin and recovery rate per 1 kg of small intestine were calculated. The residual small intestine was washed with water and then visually inspected. The results are shown below.

[0056]

[0057] As shown in Fig. 3, the residual small intestine, which was softened by water cooling after being held in a warm bath at 90 °C for 30 minutes with each extract, was softened, and a red discoloration was also observed in 1% Na ２ CO ３ , 0.1 mol / L carbonate buffer (pH 10), and 0.05 mol / L NaOH. The recovery rate of heparin from the small intestine was a maximum extraction efficiency of 17% with 0.05 mol / L NaOH, 6.7% with 0.1 mol / L carbonate buffer, and 11% with 1% Na ２ CO ３ . Therefore, although alkaline solutions such as NaOH, carbonate buffer, and Na ２ CO ３ can be used for heparin extraction, when treated at a high temperature of 90 °C, the extraction efficiency is poor, and although the shape and tissue structure of the residual small intestine remain, it is suggested that the damage to the residual small intestine such as softening and discoloration is also large, which is not preferable.

[0058] Example 5-2: Examination of Alkaline Concentration Since it was suggested from Example 5-1 that high-temperature treatment is not preferable, treatment was carried out at a low temperature, and the recovery rate of heparin and the appearance of the residual small intestine were confirmed for each alkaline concentration. The pig small intestine after washing and boiling was immersed in 3.3 times the amount of 0.1 mol / L, 0.2 mol / L, and 0.3 mol / L aqueous NaOH solutions, and heparin was extracted by performing alkaline treatment at 5 °C overnight. The extract and the residual small intestine were separated. After separation, the extract was measured in the same manner as in Examples 2 and 3, and the amount of heparin per 1 kg of small intestine and the recovery rate were calculated. The appearance of the small intestine was visually confirmed and evaluated according to the following three-level evaluation. 1: There is significant damage to the shape. 2: There is no significant damage to the shape. 3: There is little or no damage to the shape. The results are shown in Table 4 and Fig. 4 below.

[0059]

[0060] As shown in Figure 4, up to a NaOH concentration of 0.2 mol / L, there was no significant damage to the shape and tissue structure of the residual small intestine. However, when the concentration was increased to 0.3 mol / L, although the shape and tissue structure of the residual small intestine remained, the damage became significant. The results from Examples 5-1 and 5-2 suggest that, from the viewpoint of minimizing damage, an alkali concentration range of 0.05 mol / L to less than 0.3 mol / L, for example, 0.05 mol / L to 0.2 mol / L, can be adopted.

[0061] Example 5-3: Examination of extraction time Example 5-3-3: After removing the lower limit fat, washing and boiling, 90 g of pig small intestine was immersed in 3.3 times the volume of 0.1 mol / L NaOH aqueous solution, placed in a sealed container, and subjected to alkaline treatment with shaking for 2 or 6 hours to extract heparin. After the predetermined time had elapsed, 6N HCl was added to the NaOH used in equimolar amounts, neutralized with shaking, and the extract and residual small intestine were separated. After separation, the extract was measured in the same manner as in Examples 2 and 3, and the amount of heparin and recovery rate per 1 kg of small intestine were calculated. The results are shown in Table 5 below.

[0062]

[0063] As shown in Table 5, extraction was possible with a recovery rate of nearly 100% after a 6-hour treatment. Therefore, it was found that a sufficient amount of heparin can be recovered in 2 hours or more, and a considerable amount can be recovered in 6 hours or more. Thus, it can be concluded that the time required for heparin extraction is preferably 2 hours or more, and more preferably 6 hours or more.

[0064] Example 5-3-4: 150 g of porcine small intestine, after upper limit washing and boiling, was immersed in 3.3 times the volume of 0.2 mol / L NaOH aqueous solution. The solution was placed in a sealed container and subjected to alkaline treatment with shaking for 16, 24, 48, or 72 hours to extract heparin. After the predetermined time, 6N HCl was added to the NaOH used in equimolar amounts, and neutralized with shaking to separate the extract from the residual small intestine. The weight of the small intestine before extraction and the residual small intestine after extraction and separation were measured, and the increase rate was calculated. The appearance was visually confirmed.

[0065] The results are shown in Figures 5-8. As shown in these figures, the weight of the small intestine residue after extraction increased by 7.8% compared to before extraction after 16 hours of extraction, and by 21.7% after 24 hours of extraction. Furthermore, the small intestine after 16 and 24 hours of treatment had almost the same appearance, shape, and tissue structure as before extraction, although it was less swollen and the surface was slightly smoother, and lipids remained. Therefore, it can be said that the edible state is maintained even after 16 or 24 hours of heparin extraction. On the other hand, the weight increased by 79.6% after 48 hours and by 92.8% after 72 hours. In addition, the small intestine after 48 hours of treatment was swollen due to water absorption, and some of the surface was eroded. The small intestine after 72 hours of treatment showed even more severe swelling and surface erosion, and in addition, dissolution of lipids was observed. Therefore, from the viewpoint of suppressing organ damage and maintaining an edible state, it is preferable that the extraction time be less than 48 hours, and more preferably 24 hours or less. In Example 5-3-3, it was found that when a 0.1 mol / L NaOH aqueous solution was used, a sufficient amount of heparin could be recovered in 2 hours or more, and almost 100% of the heparin could be recovered in 6 hours or more. Therefore, the upper limit of the extraction time is such that a sufficient amount of heparin can be recovered, for example, 2 hours or more, 4 hours or more, preferably 6 hours or more, and any time such as 20 hours or less, 16 hours or less, or 12 hours or less can be adopted, such as a time such that damage can be suppressed and an edible state can be maintained, preferably less than 48 hours, more preferably 24 hours or less.

[0066] Example 5-3-5: Confirmation of Processing Suitability Based on the results of Example 5-3-4, even with extraction for 16 to 24 hours, the organs suffered little damage and remained in an edible state. Therefore, the processing suitability was confirmed by the following method. The small intestine extracted in Example 5-3-4 was frozen at -20°C, then thawed in a 30°C water bath, and further reboiled at 90°C for 30 minutes. Similar to Example 5-3-4, the weight of the small intestine before extraction, after thawing, and after reboiling was measured, the percentage increase was calculated, and the appearance was visually confirmed.

[0067] The results of Example 5-3-5, along with the results of Example 5-3-4, are shown in Figures 9-10. As shown in these figures, even when the residual small intestine after extraction was frozen and thawed, or subsequently reboiled, there were no significant changes in appearance, shape, tissue structure, or weight after 16 hours and 24 hours of extraction, and lipids remained. Therefore, even when heparin was extracted for 16 hours or 24 hours, the damage from processing such as freezing, thawing, and reboiling was minimal, and the edible state was maintained, indicating that the suitability for processing was preserved. On the other hand, the small intestine after 48 hours and 72 hours of processing recovered from the swollen state caused by water separation from the residual small intestine after freezing and thawing, but it had a dried-up and hard appearance and was unsuitable for consumption. After reboiling, the weight decreased even further than after thawing, the surface scratches increased, the damage was greater, and the fat content also decreased. Therefore, from the viewpoint of maintaining suitability for processing, it is preferable that the extraction time be less than 48 hours, and more preferably 24 hours or less. Therefore, for the same reasons as described above, the upper limit of the extraction time can be any time, preferably less than 48 hours, more preferably 24 hours or less, if it is 2 hours or more, 4 hours or more, preferably 6 hours or more, for example, 20 hours or less, 16 hours or less, or 12 hours or less.

[0068] Example 5-4: Temperature Investigation Example 5-1 suggested that low-temperature processing is preferable. Therefore, the temperature of the extract and residual small intestine was varied during processing, and the appearance of the small intestine was observed. Using washed and boiled pig small intestine and 0.2 mol / L NaOH (pH 13.0), the temperatures of the extract and small intestine were adjusted as shown in Table 6 below. The small intestine was immersed in 3.3 times its volume of extract, and immediately afterwards, it was left in a 5°C refrigerator or left at room temperature (approximately 25°C) for 2 hours, then left to stand in a 5°C refrigerator overnight for extraction. After extraction, 6N HCl was added to neutralize the NaOH used in equimolar amounts, and the extract and residual small intestine were separated. After separation, the residual small intestine was washed with 330 ml of water, and its appearance was visually inspected and evaluated.

[0069] The results are shown in Figure 11. It was found that when the small intestine or extract was at 20°C, damage occurred to the residual small intestine after extraction. In particular, when both the small intestine or extract was at 20°C, the damage to the residual small intestine was significant, and its shape and tissue structure could not be maintained. When either the small intestine or extract was at 20°C, the shape and tissue structure of the residual small intestine remained, but damage to the residual small intestine, such as surface dissolution, was observed. On the other hand, when both the small intestine or extract was at 8°C, there was no damage to the residual small intestine, and its shape and tissue structure remained. Therefore, it was found that it is preferable to adjust the material to a low temperature beforehand before performing heparin extraction in order to maintain the physical properties of the small intestine.

Claims

1. Edible organs characterized by having a heparin content of 10 mg / kg or less per organ.

2. A method for extracting heparin, characterized by comprising the step of extracting heparin from an organ obtained from a non-human animal by alkaline treatment at a temperature of 10°C or lower.

3. A method for producing edible organs, characterized by comprising the step of producing edible organs by alkaline treatment of organs obtained from non-human animals at a temperature of 10°C or lower.

4. A method for processing organs, comprising the steps of: providing heparin, which includes extracting heparin from organs obtained from non-human animals by alkaline treatment at a temperature of 10°C or lower; and providing edible organs, which includes obtaining the organs after the alkaline treatment as edible organs.

5. The method according to claim 3 or 4, characterized in that the heparin content in the edible organs obtained by the above method is 10 mg / kg or less per organ.

6. An organ processing system characterized by comprising: a heparin providing means for extracting and providing heparin from organs obtained from non-human animals by alkaline treatment at a temperature of 10°C or lower; and an edible organ providing means for obtaining the organs after the alkaline treatment as edible organs.

7. The method described in claim 3 or 4 or An edible organ obtained by the system of claim 6.

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