JP5553325B2 - Natural polymer compound extraction and purification system - Google Patents

Description

  The present invention relates to a system for extracting and purifying natural polymers (eg, polysaccharides, proteins) from natural materials.

  Polysaccharides play various roles in the body. Among these, mucopolysaccharides such as chondroitin sulfate, hyaluronic acid, and heparin are recently attracting attention. For example, chondroitin sulfate has been recognized for its effects on arthritis and neuralgia, and is used in health foods, supplements and pharmaceuticals. Hyaluronic acid is often used in cosmetics and the like because it has a moisture retention effect. Heparin has an anticoagulant action and is used in the medical field. These substances have been conventionally used as pharmaceuticals, and recently, the demand for health foods and supplements has increased dramatically. These polysaccharides are obtained using natural tissue as a raw material. Many methods have been developed, but it is not efficient because it takes time especially for the dialysis step, ultrafiltration step, precipitation step with organic solvent, etc., and it is large, complicated and expensive. Many facilities are required (see Patent Documents 1 to 3).

  In addition, useful polysaccharides such as mucopolysaccharides are mostly derived from animal (including terrestrial and aquatic) and fungal tissues, and in most cases are obtained from animal tissues that humans eat. Yes. Recently, from the viewpoint of effective use of resources, attempts have been made to obtain useful polysaccharides from inedible parts (parts not suitable for cooking) of food animals that have been discarded. Moreover, the amount of inedible parts of edible animals is enormous.

In view of such circumstances, there is a need for a method for obtaining a large amount of useful polysaccharides from a large amount of raw materials in a short time by a simple and efficient method.
JP 2000-273102 A JP 2003-268004 A JP 2004-189825 A

  An object of the present invention is to provide a system for automatically acquiring a large amount of natural polymer (useful polysaccharide) from a large amount of natural raw material in a short time.

Specifically, the extraction / purification system (1) according to the present invention extracts the natural polymer compound contained in the natural material (160) while stirring the mixture of the natural material (160) and the extraction solvent (162). An extraction device (2) and a purification device (4) for purifying the natural polymer compound extracted by the extraction device (2) are provided. The extraction device (2) includes an outer tub (10) for containing the extraction solvent (162) and a plurality of communication holes (64) through which the extraction solvent (162) can pass, and contains the natural material (160). The inner tank (46) fixed concentrically with the outer tank (10) inside the outer tank (10) and the inner tank (46) fixed to the outer tank (10) are reversed. A rotating means (40) for rotating and an opening ratio adjusting means (66) for adjusting the opening ratio of the inner tank (46) are provided. The rotating means (40) includes a motor drive shaft (42) protruding inward of the outer tub (10) and detachably connected to the inner tub (46) . The opening ratio adjusting means (66) comprises (a) the inner tub (46) comprising an outer basket (48) and an inner basket (50) disposed inside the outer basket (48), b) A plurality of outer openings (56) are formed in the outer basket (48), a plurality of inner openings (62) are formed in the inner basket (50), and (c) the inner basket (50) is formed. In a state of being arranged inside the outer basket (48), at least a part of the outer opening (62) overlaps at least a part of the inner opening (56) to form the plurality of communication holes (64). And (d) the opening ratio of the inner tub (50) can be adjusted by changing the relative position of the inner basket (50) with respect to the outer basket (48). Location (4) comprises a column (100) for gel filtration chromatography.

  According to the extraction / purification system having such a configuration, a large amount of useful polysaccharides can be obtained from a large amount of raw materials in a short time simply and efficiently.

  Hereinafter, an embodiment of a natural polymer compound (polysaccharide) extraction / purification system according to the present invention will be described with reference to the drawings.

[1. System overview)
As shown in FIG. 1, the extraction / purification system 1 generally includes an extraction device 2 that extracts a natural polymer (for example, polysaccharide) from a natural material using an extraction solvent, and a natural high content extracted by the extraction device 2. A purification device 4 for purifying the molecule, a transport device 6 for transporting the extraction solvent and the natural polymer from the extraction device 2 to the purification device 4, and an automatic control device 8 for automatically controlling the extraction device 2 and the transport device 6 are provided.

[2. Extraction device)
Referring to FIGS. 2 and 3, the extraction device 2 has an outer tub (constant temperature bath) 10. In the embodiment, the outer tub 10 is a cylindrical container having an opening at the top, and is fixed to a floor 12 of a building (not shown). As shown in detail in FIG. 2, the outer tub 10 includes an outer cylindrical portion 16 and an inner cylindrical portion 18 that are arranged on a circumference around the central axis 14, and a bottom portion of the outer cylindrical portion 16 and the inner cylindrical portion 18. Bottom plates 20 and 22, respectively, and an annular plate portion 24 connecting the upper ends of the outer cylindrical portion 16 and the inner cylindrical portion 18, and a hot water circulation chamber (temperature adjustment medium) in a space surrounded by these members. The fluid which is the temperature adjusting medium 28 can be sealed and accommodated in the (accommodating chamber) 26.

  The temperature of the temperature adjusting medium 28 accommodated in the hot water circulation chamber 26 is optimally adjusted according to the natural material to be processed. Therefore, the extraction device 2 according to the embodiment includes a temperature detector 30 that detects the temperature of the temperature adjustment medium 28 in the hot water circulation chamber 26, and temperature adjustment if necessary according to the temperature detected by the temperature detector 30. A heater 32 for heating the medium 28 and circulation means 34 for circulating the temperature adjustment medium 28 in the hot water circulation chamber 26 are provided. For example, as shown in FIGS. 1 and 2, the circulation means 34 includes a circulation pipe 36 that connects the bottom and the top of the hot water circulation chamber 26, and a pump 38 provided in the circulation pipe 36, and drives the pump 38. As a result, the temperature adjusting medium 28 in the hot water circulation chamber 26 is circulated through the circulation pipe 36.

  A motor 40 that can rotate forward and reverse is fixed to the bottom of the outer tub 10. The drive shaft 42 of the motor 40 is arranged coaxially with the central axis 14 of the outer tub 10, passes through the bottom plates 20 and 22 of the outer tub 10, and protrudes inside the outer tub 10. As shown in the drawing, the periphery of the drive shaft 42 penetrating the hot water circulation chamber 26 is surrounded and sealed by a cylindrical or annular partition wall 44.

  An inner tank (material storage tank) 46 is detachably connected to a motor drive shaft 42 protruding to the inside of the outer tank 10. As shown in the drawing, the inner tank 46 connected to the motor drive shaft 42 is formed of a cylindrical body, and the central axis of the cylindrical body is made to coincide with the central axis 14 of the outer tank 10. As shown in detail in FIG. 3, the inner tub 46 includes a cylindrical outer basket 48 and an inner basket 50. The outer basket 48 is in the form of a cylindrical container with an open top, and has a cylindrical portion 52 and a bottom portion 54. The cylindrical portion 52 and the bottom portion 54 regularly have a large number of openings (outside openings) 56 that communicate between the inside and the outside. Similarly, the inner basket 50 is in the shape of a cylindrical container with an open top and has a cylindrical portion 58 and a bottom portion 60. The cylindrical portion 58 and the bottom portion 60 regularly have a large number of openings (inner openings) 62 that communicate between the inside and the outside.

  The inner diameter of the outer basket cylindrical portion 52 is substantially equal to the outer diameter of the inner basket cylindrical portion 58. Accordingly, when the inner basket 50 is inserted into the outer basket 48, the outer surface (outer cylindrical surface and outer bottom surface) of the inner basket 50 is flush with or substantially flush with the inner surface (inner circumferential cylindrical surface and inner bottom surface) of the outer basket 48. In addition, a substantial gap is not formed between the cylindrical portions 52 and 58. The cylindrical portions 52 and 58 are substantially equal in height, and in a state where the inner basket 50 is inserted in the outer basket 48, the upper ends of both coincide with each other, or the upper end of the inner basket 50 is slightly higher than the upper end of the outer basket 48. It is preferable to protrude.

  In the embodiment, the shapes of the openings 56 and 62 formed in the outer and inner baskets 48 and 50 are circular, but the shape is arbitrary (for example, a square shape, other rectangular shapes, a slot shape, a star shape). Specifically, a known stainless steel punching metal can be suitably used as a material constituting the baskets 48 and 50 (cylindrical portion and bottom plate). As is well known, the punching metal has openings 56 and 62 regularly formed in the baskets 48 and 50 at regular intervals in the vertical direction, the horizontal direction, and the diagonal direction. Accordingly, when the inner basket 50 is rotated or moved up and down relative to the outer basket 48, the opening 56 of the outer basket 48 and the opening 62 of the inner basket 50 are formed to coincide with each other. The area (opening area or opening ratio of the inner tank) changes.

  It is preferable to include an aperture ratio adjusting means 66 that can adjust the aperture ratio of the inner tank 46 to a plurality of predetermined values. In the embodiment, a method of adjusting the aperture ratio by rotating the inner basket 50 with respect to the outer basket 48 is employed. Therefore, as shown in FIG. 6, in the embodiment, as the opening ratio adjusting means 66, the annular plates 68 and 70 are fixed to the center of the basket bottom portions 54 and 60, and one annular plate 70 of the inner basket 50 is provided. A through hole 72 is formed, and a plurality of screw holes 74 a to 74 e are formed in the annular plate 68 of the outer basket 48, and a plurality of screws of the outer basket annular plate 68 are passed through the through holes 72 of the inner basket annular plate 70. The opening ratio can be adjusted to 100%, 80%, 70%, 60%, and 50% by screwing the screw 76 into any of the holes 74a to 74e.

  Instead, the opening ratio can be adjusted by raising the inner basket 50 relative to the outer basket 48. In this case, although not shown, means (for example, a height adjusting jig, a bolt and screw hole, or a combination of a bolt and a nut) capable of fixing the inner basket 50 at a plurality of positions with respect to the outer basket 48 in the height direction is provided. The height of the inner basket 50 is adjusted according to the required opening ratio.

  Different openings may be provided in the outer and inner baskets 48,50. The openings 56 and 62 are not necessarily formed in the entire baskets 48 and 50, and may be formed only in at least a part of the baskets 48 and 50 as long as the opening ratio of the inner tank 46 can be adjusted. Further, when adjusting the opening ratio, when the inner basket 50 is rotated or raised with respect to the outer basket 48, the overlapping area of all the openings 56, 62 need not change, and at least one of the openings 56, 62 does not change. What is necessary is just to make it the aperture ratio of the inner tank 46 change as a result by changing the overlapping area of a part.

  2 and 4, the inner basket 50 is preferably provided with a stirring blade 78 on the inner surface of the cylindrical portion 58, the inner surface of the bottom portion 60, or both. The shape and size of the stirring blade 78 can be appropriately changed according to the type of natural material to be processed.

  As shown in FIGS. 2 and 3, in order to accurately and concentrically position the inner tub 46 with respect to the outer tub 10, it is preferable that guide means 80 be arranged uniformly in the vicinity of the upper end of the outer tub 10. For example, in the embodiment, each guide means 80 includes a bracket 82 fixed to the inner surface of the outer tub 10 and a roller 86 rotatably supported via a vertical shaft 84 fixed to the bracket 82. Of course, in order to accurately position the inner tub 46 inside the plurality of rollers 86, a circle (not shown) inscribed in the plurality of rollers 86 may coincide with or substantially coincide with the outer peripheral surface of the inner tub 46. preferable.

  Referring to FIG. 2, the inner tank 46 configured as described above is inserted into the outer tank 10. At this time, the motor drive shaft 42 is inserted into a central through hole 88 (see FIG. 6) of the annular plate 70 provided at the center of the inner basket bottom portion 60, and an upper end external screw portion (not shown) of the motor drive shaft 42 is inserted. The inner basket 50 is fixed to the motor drive shaft 42 by screwing a cap 90 having an inner thread. As shown in the drawing, in this state, a space with a substantially constant thickness is formed between the outer tub 10 and the inner tub 46, and around the motor drive shaft 42 protruding into the outer tub 10. An annular adjusting member 92 is preferably provided.

[3. (Purification equipment)
Returning to FIG. 1, the purification device 4 has a column 100 for gel filtration chromatography. As will be described later, a column 102 for ion exchange chromatography may be additionally provided. Since the configuration of the columns 100 and 102 is well known, a description thereof will be omitted.

[4. (Transport equipment)
The transport device 6 is a pipe that connects the inside of the outer tank 10, more precisely, the bottom of the space (processing chamber) surrounded by the inner cylindrical portion 18 and the inner bottom plate 22 and the upper portion (inlet) of the column 100 for gel filtration chromatography. 104, a pipe 106 that connects the lower end (outlet) of the column 100 for gel filtration chromatography and the upper end (inlet) of the column 102 for ion exchange chromatography. The former pipe 104 is provided with two solenoid valves 108 and 110 and one pump 112. The latter pipe 1096 is provided with two solenoid valves 114 and 116 and one pump 118. The lower end (exit) of the ion exchange chromatography column 102 and the electromagnetic valve 116 are connected by a pipe 120, and another electromagnetic valve 122 is provided in the pipe 120. And solenoid valve 108,110,114,122 is connected to the container 132,134,136,138 corresponding to piping 124,126,128,130, respectively.

[5. Automatic control unit)
The automatic control device 8 includes a central processing unit (CPU) 140, a ROM 142, a RAM 144, a timer 146, an input unit 148, and an output unit 150. The configuration and operation of each unit will be specifically described in the operation of the system described below.

[6. System operation)
A method for obtaining a polysaccharide, which is a polymer compound, from a natural material using the system 1 having the above configuration will be described. Examples of natural tissues rich in useful polysaccharides include those derived from mycelium and fruit bodies of basidiomycetes such as Agaricus, cartilage and internal organs of fish and animals, and the head of squid. For example, chondroitin sulfate can be obtained from fish (including ray and shark) cartilage. Hyaluronic acid can be obtained from chicken tosaka and the like. Alternatively, heparin can be obtained from porcine small intestinal mucosa. In addition, inedible parts of edible organisms, for example, livestock such as cattle, pigs and chickens, fish such as salmon and sardines, molluscs such as squid and octopus, shellfish such as shrimps and crabs, etc. It is enormous and useful polysaccharides can be obtained from it. Polysaccharides can also be obtained from tissues such as jellyfish. Further, for example, mucopolysaccharides such as chondroitin sulfate, hyaluronic acid and heparin are included. For example, chondroitin sulfate can be obtained from fish (including ray and shark) cartilage, hyaluronic acid can be obtained from chicken crest, etc., or heparin can be obtained from porcine small intestinal mucosa. In addition, for example, squid, such as green squid, squid, cuttlefish, squid, squid, squid, squid, squid, squid, squid, squid, etc. Other inedible parts), cattle, pigs, horses, goats, sheep and other livestock and poultry organs (eg liver, intestines, airways) and other parts (eg nasal cartilage cartilage, eyeballs, skin, etc.) In particular, chondroitin sulfate and other polysaccharides can also be obtained from chicken cartilage (eg breast cartilage, articular cartilage, etc.) or fish cartilage. Furthermore, for example, polysaccharides such as beta glucan can be obtained from basidiomycetes such as Agaricus and Sarnococcus.

[6-1. Preparation process)
In the extraction device 2, the temperature adjustment medium 28 is filled in the hot water circulation chamber 26 of the outer tub 10. As the temperature adjusting medium 28, water or ethylene glycol can be suitably used. Further, the inner tank 46 is charged into the outer tank 10, and the inner tank 46 is fixed to the outer tank 10. At this time, the inner tub 46 fixes the position of the inner basket 50 with respect to the outer basket 48 according to a desired opening ratio. Next, a natural material (processing material) 160 to be processed is put into the inner tank 46. The size of the natural material 160 to be charged is larger than the communication hole (opening area) 64 of the inner tank 46 and the openings 56 and 62 of the baskets 48 and 50, and the natural material 160 is externally connected through the communication hole 64 and the openings 56 and 62. Will not be leaked. Further, the extraction solvent 162 is put inside the inner tank 46 and the outer tank 10. As the extraction solvent 162, an aqueous solvent is preferably used. For example, water, a buffer near neutrality (for example, an acetate buffer having a pH of about 5 to about 9, a potassium phosphate buffer, a sodium phosphate buffer, etc.) is used. Although illustrated, it is not restricted to these. Alternatively, a mixture of an aqueous solvent and an organic solvent (for example, ethanol, acetone, etc.) may be used for extraction.

  In the purification device 4, the gel filtration chromatography column 100 is filled with a gel filtration carrier. Conditions such as the type of carrier used for gel filtration, column size, eluate composition, flow rate, etc. are factors such as the origin of living tissue, the type of polysaccharide required (and molecular weight, charged state, etc.), quantity, and purity. Depending on the situation, those skilled in the art can appropriately select and determine. In selecting and determining these conditions, the necessary polysaccharides are eluted in the flow-through fraction or polymer fraction with little or no trapping on the gel filtration support, and other components are added to the gel filtration support. Be trapped. Examples of the gel filtration carrier used in the method of the present invention include dextran, agarose, polyacrylamide, and glass beads. Specific examples include various types of Sepharose, various types of Sephadex, various types of Sephacryl, various types of Toyopearl, various types of Superdex, various types of Superrose, and the like. Moreover, the developing solution used for the gel filtration process is filled in the container 134. Examples of the developing solution include, but are not limited to, water, a neutral buffer solution (for example, an acetate buffer solution having a pH of about 5 to about 9, a potassium phosphate buffer solution, a sodium phosphate buffer solution, and the like). Further, a salt such as sodium chloride may be added to the developing solution in order to adjust the interaction between the gel filtration carrier and the polysaccharide.

  Further, the ion exchange chromatography column 102 is filled with an ion exchanger. The ion exchanger can be appropriately selected in consideration of the type of polysaccharide required, the required purity, the type and amount of raw material, the required yield of polysaccharide, and the like. Various ion exchangers are commercially available, and can be selected from them. Examples of anion exchangers include anion exchange resins such as Dowex, Amberlite and Muromac, hydrotalcite, chitopearl “basic”, DEAE-chitopearl, Capto Q, Mono Q, SAX, and Exe. Examples include anion-exchange cellulose such as DEAE-cellulose and DEAE-Sephadex, which are also commercially available. A weak anion exchanger can be used for separation and purification of a polysaccharide having a sulfate group, and for example, an anion exchanger such as DEAE-cellulose, DEAE-Sephadex, and Dowex can be used. In order to purify the target polysaccharide in the ion exchange chromatography step, the target polysaccharide is usually adsorbed on the ion exchanger, and then the ionic strength of the eluent is increased. The desired polysaccharide is eluted from the ion exchanger. Examples of the method for increasing the ionic strength include a stepwise method and a gradient method. Moreover, the purity of the target polysaccharide can be further increased by using a chromatography step with an ion exchanger having an opposite charge that does not adsorb the target polysaccharide. For this purpose, a cation exchanger is preferably used. Examples of the cation exchanger include cation exchange resins such as Dowex, Amberlite and Muromac, and cation exchangers such as zeolite, carboxylated chitopearl, sulfonated chitopearl, Capto S, Mono S, SCX, and Exe. It is also commercially available. The ion exchange chromatography step used in the present invention may be performed by either a column type or a batch type. Selection of the ion exchanger used in the ion exchange chromatography process, the type of eluent, the composition, the concentration of each component, pH, temperature, flow rate, and other conditions are within the normal range of ordinary skill in the art. .

[6-2. (Extraction process)
Depending on the natural material (process material) 160 to be processed, the set temperature of the temperature adjusting medium 28 stored in the hot water circulation chamber 26 of the outer tub 10 and the forward / reverse rotation time of the motor 40 through the input unit 148 in the control device 8. The drive time of the motor 40 is set (rotation time in the reverse direction with respect to the forward rotation time). The set conditions are stored in the RAM 144. The set temperature, rotation time, and drive time are determined according to each processing material, the determined setting temperature and the like are stored in the ROM 142 of the control device 8, and the type of the processing material is simply input from the input unit 148. The set temperature or the like may be set automatically.

  When the input of necessary conditions is completed, the CPU 140 of the control device 8 drives the heater 32 and the circulation pump 38 via the output unit 150 based on the input conditions, and the hot water circulation chamber 26 of the outer tub 10. While the temperature adjusting medium 28 is circulated through the circulation pipe 36, the temperature of the temperature adjusting medium 28 is adjusted based on the output of the temperature detector 30 (the temperature of the temperature adjusting medium 28). Note that the temperature of the temperature adjustment medium 28 in the extraction process does not need to be constant. For example, the set temperature may be changed at each stage based on a temperature management program (not shown) input to the ROM 142 or the RAM 144. .

  The CPU 140 also switches the rotation direction of the motor 40 alternately between the forward direction and the reverse direction, and rotates the inner tank 46 alternately in the forward direction and the reverse direction. Thereby, the natural material 160 accommodated in the inner tank 46 is stirred by the stirring blade 78 provided in the inner tank 46 and is sufficiently brought into contact with the extraction solvent 162, so that the natural polymer is extracted well.

  The rotation time in the forward direction and the rotation time in the reverse direction (rotation duration) can be arbitrarily determined. For example, it can be input through the input unit 148, or a rotation time suitable for the processing material can be stored in the ROM 142 in advance, and the CPU 140 can control the driving of the motor 40 based on the information. A predetermined pause time may be provided between the rotation in the forward direction (or the reverse direction) and the subsequent rotation in the reverse direction (or the forward direction). In this case, first, the extraction solvent 162 in the inner tank 46 is moved radially outward by centrifugal force due to rotation in the forward direction (or the reverse direction), and is moved to the outside of the inner tank 46 through the communication hole 64 of the inner tank 46. Moving. Similarly, the processing material also receives centrifugal force and gathers in the vicinity of the peripheral wall of the inner tank 46. Next, when the rotation in the forward direction (or the reverse direction) is stopped, the extraction solvent 162 that has moved to the outside of the inner tank 46 moves to the inside of the inner tank 46 through the communication hole 64 of the inner tank 46. As a result, the movement of the extraction solvent 162 disperses the natural material 160 gathered in the vicinity of the peripheral wall of the inner tank 46, and each processing material piece effectively comes into contact with the extraction solvent 162, thereby promoting the extraction of the natural polymer. Is done.

  Stirring of the natural material 160 and the extraction solvent 162 is performed for a predetermined time. This time is a time set by the input unit 148. Instead, the rotation time suitable for the processing material may be stored in the ROM 142 in advance, and the CPU 140 may control the driving time (total stirring time) of the motor 40 based on the information.

  The stirring time varies depending on the natural material and the natural polymer to be extracted. For reference, when chondroitin sulfate is extracted from squid skin, it is stirred for about 12 hours while maintaining the extraction solvent at room temperature. When extracting collagen from the scale, the mixture is stirred for about 2 to 3 hours while maintaining the extraction solvent at about 100 ° C. When extracting beta-glucan from shiitake, it is stirred for about 12 hours while maintaining the extraction solvent at about 100 ° C.

[6-3. (Discharge / drying process)
When stirring for a predetermined time is completed, the CPU 140 stops the motor 40. Next, the solenoid valves 108 and 110 are operated, the pump 112 is driven, and the extraction solvent 162 containing the natural polymer is sent to the column 100 for gel filtration chromatography. In order to completely discharge the natural polymer remaining between the natural materials 160, it is preferable to maintain the rotation of the motor 40 during the discharge of the extraction solvent 162. At this time, it is more preferable that the rotation speed of the motor 40 is set larger than the rotation speed of the motor 40 in the extraction step, and the inner tank 46 is rotated at a high speed. The high speed rotation time may be input from the input unit 148 or may be stored in the ROM 142 in advance. After the discharge is completed, the CPU 140 can drive the motor 40 for a predetermined time to dry the material remaining in the inner tank 46. When the extraction solvent 162 containing the extracted natural polymer is completely sent to the column 100, the CPU 140 stops the pump 112 and closes the solenoid valves 108 and 110. Thereafter, when the outer tub 10 and the inner tub 46 are cleaned, the electromagnetic valve 108 is operated to guide the cleaning water to the waste water container 132.

[6-4. Purification (gel filtration chromatography) step]
In the purification (gel filtration chromatography) step, the extraction solvent 162 and the natural polymer sent to the column 100 are subjected to gel filtration chromatography to obtain a pass-through fraction or a polymer fraction. At this time, the CPU 140 operates the electromagnetic valve 110, drives the pump 112, and supplies the developing solution from the container 134 to the column 100. As a result, in the column 100, the necessary polysaccharide is eluted in the pass-through fraction or polymer fraction with little or no trapping on the gel filtration carrier, and other components are trapped on the gel filtration carrier. Is done.

  In the selection of the chromatography conditions, more preferably, the target polysaccharide is more preferably eluted in a fraction of about 13 to about 33% of the total bed volume of the gel filtration carrier after the start of liquid feeding. Gel filtration chromatography conditions are selected to elute in fractions up to about 20% of the total bed volume of the gel filtration support. Therefore, in the present specification, the “through fraction or polymer fraction” refers to the viscosity of the sample, the type of polysaccharide contained in the sample, the molecular weight of the desired polysaccharide, the type of carrier used, etc. Refers to the fraction obtained in the eluate of up to about 33% of the total bed volume. For example, the fraction obtained in the eluate of about 13% to about 20% of the total bed volume of the gel filtration carrier may be collected as a “through-through fraction or polymer fraction” to obtain a polysaccharide.

  Various known methods can be used for detection and confirmation of polysaccharides in fractions from gel filtration chromatography, for example, carbazole method, dimethylphenol method, phenol sulfuric acid method, ultraviolet absorption measurement method, refractive index measurement. A method or the like may be used.

  During the purification (gel filtration chromatography) step, the CPU 140 opens the electromagnetic valve 114 and collects the natural polymer that has passed through the column 100 into the container 136. However, when performing the purification (ion exchange chromatography) step described below, the CPU 140 operates the solenoid valves 114 and 116 and drives the pump 118 to further use the natural polymer from the column 100 for ion exchange chromatography. Send to column 102.

  In experiments conducted by the present inventors, 100% pure chondroitin sulfate was obtained from chicken breast cartilage and 68% pure chondroitin sulfate was obtained from squid skin by gel filtration chromatography.

[6-5. Purification (ion exchange chromatography) process]
In the purification (ion exchange chromatography) step, in order to purify the target polysaccharide, the target polysaccharide is usually adsorbed to the ion exchanger in the column 102, and then, for example, the ionic strength of the eluent is increased. The target polysaccharide is eluted from the ion exchanger by the method described above. In experiments conducted by the present inventors, the purity of crude chondroitin sulfate derived from chicken knee cartilage increased from 40% to 100%.

  The CPU 140 operates the solenoid valves 114, 116, and 122 to guide the high purity natural polymer eluted from the ion exchanger to the container 136.

  When the columns 100 and 102 are washed after the purification process is accommodated, the electromagnetic valve 122 is operated based on a command from the input unit 148 to guide the washing liquid (waste water) to the container 138.

  Thus, according to the extraction / purification system 1 described above, the operation of all devices (motor 40, pump 38, solenoid valve) can be automated by the control device 8. In particular, since gel filtration uses a difference in molecular weight and does not require a regeneration step, the process from extraction to purification (gel filtration chromatography) can be completely automated, so a large amount of natural samples can be obtained almost continuously. A large amount of natural polymer can be obtained by treatment. Moreover, optimal extraction / purification conditions can be obtained according to the type of natural material.

  Moreover, when high purity is not required like a foodstuff or a food additive, it can fully respond only by gel filtration refinement | purification. On the other hand, when high purity is required like a pharmaceutical product, it can be purified to a high-purity product in combination with an ion exchange column.

  Furthermore, the above-described extraction / purification system can be used not only for polysaccharide extraction but also for protein extraction. In this case, the steps from extraction to purification can be performed automatically and continuously as described above. it can.

  Furthermore, in the above-described extraction / purification system, the inner tank 46 is composed of two baskets (the outer basket 48 and the inner basket 50), but an extraction / purification system composed of one basket is also included in the present invention.

The figure which shows schematic structure of the extraction / purification system which concerns on this invention. Sectional drawing of an extracting device. The top view of an extracting device. The disassembled perspective view of the inner tank of the extraction apparatus shown in FIG. The partial expanded sectional view of the inner tank shown in FIG. The perspective view which shows the means to adjust the aperture ratio of the inner tank shown in FIG.

Explanation of symbols

1: Extraction / purification system 2: Extraction device 4: Purification device 6: Transport device 8: Automatic control device 10: Outer tub 12: Floor 14: Center shaft 16: Outer cylindrical portion 18: Inner cylindrical portion 20: Bottom plate 22: Bottom plate 24: annular plate part 26: internal space 28: temperature adjusting medium 30: temperature detector 32: heater 34: circulation means 36: circulation pipe 38: pump 40: motor 42: drive shaft 44: partition 46: inner tank 48: Outer basket 50: Inner basket 52: Cylindrical portion 54: Bottom portion 56: Outer opening 58: Cylindrical portion 60: Bottom portion 62: Inner opening 64: Communication hole 66: Opening ratio adjusting means 68: Annular plate 70: Annular plate 78: Stirring blade 80: Guide means 140: CPU
160: Natural material 162: Extraction solvent

Claims (8)

The extraction device (2) for extracting the natural polymer compound contained in the natural material (160) while stirring the mixture of the natural material (160) and the extraction solvent (162), and the extraction device (2) An extraction / purification system comprising a purification device (4) for purifying a natural polymer compound, wherein the extraction device (2) comprises:
An outer tank (10) containing the extraction solvent (162);
It has a plurality of communication holes (64) through which the extraction solvent (162) can pass, accommodates the natural material (160), and is concentrically fixed to the outer tank (10) inside the outer tank (10). An inner tank (46) to be
Rotating means (40) for rotating the inner tank (46) fixed to the outer tank (10) forward and backward ,
An opening ratio adjusting means (66) for adjusting the opening ratio of the inner tank (46);
The rotating means (40) includes a motor drive shaft (42) protruding inward of the outer tub (10) and detachably connected to the inner tub (46) .
The aperture ratio adjusting means (66)
(A) The inner tank (46) is composed of an outer basket (48) and an inner basket (50) disposed inside the outer basket (48),
(B) forming a plurality of outer openings (56) in the outer basket (48), forming a plurality of inner openings (62) in the inner basket (50);
(C) With the inner basket (50) disposed inside the outer basket (48), at least a part of the outer opening (62) overlaps at least a part of the inner opening (56), Communication hole (64) is formed,
(D) The opening ratio of the inner tub (50) can be adjusted by changing the relative position of the inner basket (50) with respect to the outer basket (48).
The said refiner | purifier (4) is equipped with the column (100) for gel filtration chromatography, The extraction and purification system characterized by the above-mentioned . The aperture ratio adjusting means (66) rotates the inner basket (50) relative to the outer basket (48) around the rotation axis (42) of the rotating means (40), The extraction / purification system according to claim 1, wherein the opening ratio of the inner tank (46) is adjustable. The aperture ratio adjusting means (66) moves the inner basket (50) relative to the outer basket (48) along the central axis of the rotating shaft (42) of the rotating means (40). The extraction / purification system according to claim 1, wherein the opening ratio of the inner tank (46) can be adjusted. The opening ratio adjusting means (66) is a fixing means (74a to 74e, 76) for fixing the inner basket (50) to the outer basket (48) corresponding to a plurality of predetermined opening ratios. The extraction / purification system according to claim 2 or 3, further comprising: The extraction / purification system includes a control means (8),
The said control means (8) is equipped with the adjustment means (140,142,148) which adjusts the time which rotates the said inner tank (46) in the forward direction and the reverse direction. Item 5. The extraction / purification system according to any one of Items 1 to 4. The extraction device also adjusts the temperature of the extraction solvent contained in the outer tank.
A temperature control medium accommodation chamber (26) formed in the outer tub;
A medium (28) accommodated in the temperature adjusting medium accommodation chamber (26);
A circulating means (38) for circulating the medium through the temperature adjusting medium storage chamber (26);
A temperature detector (30) for detecting the temperature of the medium (28);
A heater (32) for heating the medium (28);
The extraction / purification system according to any one of claims 1 to 5, further comprising a heater control means (140) for controlling the heater (32). The above extraction and purification system is also
The extraction device (2) and the purification device (4) are connected to transport the extraction solvent (162) containing the natural polymer compound extracted by the extraction device (2) to the purification device (4). Transport piping (104);
A pump (112) provided in the transport pipe (104);
After the extraction process in the extraction device (2) is completed, the pump (112) is driven to automatically transport the extraction solvent (162) from the extraction device (2) to the purification device (4). The extraction / purification system according to any one of claims 1 to 6, further comprising transport control means (140). The extraction / purification system according to any one of claims 1 to 7, wherein the purification device (4) has an ion exchange chromatography column (102).

Images