JP4431009B2 - Supercritical fluid chromatography column - Google Patents

Description

  The present invention relates to a column used for supercritical fluid chromatography, and particularly to a column suitably used for fractionation of a target substance in supercritical fluid chromatography.

  Supercritical fluid chromatography is usually performed in a high pressure system several to several tens of times higher than that of high performance liquid chromatography. Accordingly, the column used for supercritical fluid chromatography is required to have higher pressure resistance and air tightness than the column used for high performance liquid chromatography.

  In supercritical fluid chromatography, a supercritical fluid having excellent diffusibility and low viscosity is used for the mobile phase. For this reason, supercritical fluid chromatography is used for fractionation of substances that are usually difficult to separate, such as optical isomers.

  As a column used for the supercritical fluid, for example, a separation tube for containing a separation agent, a carrier tube containing the separation tube, and an adapter provided at the end of the carrier tube to hold the separation tube in the carrier tube There is known a column including a PEEK sleeve for fitting the adapter member to the end of the separation tube and closing the gap between the separation tube and the carrier tube (for example, Patent Documents). 1).

  This column is a so-called packed column, and is excellent from the viewpoint of enhancing versatility mainly in analytical applications, but there remains room for examination as a preparative column.

  In general, a column having a large inner diameter is suitably used for fractionation of substances from the viewpoint of increasing the fractionation efficiency. For this reason, the column for fractionation has a problem that it is difficult to keep the packed state of the particulate separation agent packed normally. As a technique for solving such a problem, a movable stopper that is movable in the hollow tube along the axial direction of the hollow tube is employed as at least one of the stoppers provided at the end of the hollow tube. A movable stopper type column is known.

  The movable stopper type column is, for example, a movable stopper type liquid chromatograph in which a movable stopper having an openable / closable flow path is slidably inserted into a separating agent filling shell having an openable / closable flow path at one end. In a chromatography column, a movable stopper-type liquid chromatograph characterized in that a nut-shaped hub is screwed onto a bolt-shaped piston rod connected to the back of a movable stopper, and the hub is fixed to a flange of a shell by a locking means. A chromatography column is known (for example, see Patent Document 2).

  In addition, the movable stopper column is, for example, a column used in a simulated moving bed chromatography separation apparatus, in which one of both ends of the column is a movable stopper and the other is a stationary stopper. A column for a bed type chromatography separation apparatus is known (for example, refer to Patent Document 3).

The above-described movable stopper column can be fixed in a state where the movable stopper is pressed toward the separating agent accommodated in the hollow tube, and is excellent from the viewpoint of keeping the separating agent packed normally. However, in supercritical fluid chromatography, a supercritical fluid excellent in diffusibility and low viscosity is used for the mobile phase, and the pressure of the mobile phase is high. In the case of using for the supercritical fluid in the mobile phase may leak from the movable stopper.
Japanese Patent Laid-Open No. 9-111992 JP-A-8-43373 JP-A-8-155207

  An object of the present invention is to provide a column capable of preventing leakage of a mobile phase and maintaining a packed state of a particulate separating agent in supercritical fluid chromatography.

  The present invention uses a seal for preventing leakage of a supercritical fluid in addition to a separating agent and a solvent, as a seal provided on the movable stopper.

  That is, the present invention includes a hollow tube for containing a separating agent for separating a target substance, and a stopper provided at both ends of the hollow tube for allowing a fluid to flow without leaking the separating agent. In a column used for supercritical fluid chromatography using a fluid containing a supercritical fluid and a solvent as a mobile phase, at least one of the stoppers is movable in the hollow tube along the axial direction of the hollow tube The column further includes a fixing means for fixing the movable plug in a pressed state along the axial direction of the hollow tube, and when the movable plug is accommodated in the hollow tube, A fluid flow path that penetrates the movable stopper along the axial direction of the empty pipe, and a seal that closes a gap between the movable stopper and the inner peripheral wall of the hollow pipe, and the seal is in the axial direction of the hollow pipe. Including a first seal and a second seal, the first seal being divided Agent and a seal to prevent leakage of the solvent, the second seal provides a column is a seal to prevent leakage of the supercritical fluid.

  According to the above configuration, in the movable range of the movable stopper, it is possible to press the separating agent accommodated in the hollow tube and fix the movable stopper in that state. Can be kept normal. Moreover, according to the said structure, leakage of the supercritical fluid from a movable stopper is prevented by said 2nd seal, and it becomes possible to prevent the leakage of a mobile phase.

  In the present invention, the hollow tube further includes a first screw provided around the outer peripheral surface of the end portion of the hollow tube, and the fixing means includes a peripheral wall portion surrounding the outer peripheral surface of the hollow tube, It is preferable to have a second screw provided on the inner peripheral wall of the peripheral wall portion and screwed into the first screw, and a flange portion extending from the inner peripheral wall of the peripheral wall portion toward the central axis of the peripheral wall portion. According to such a configuration, the flange portion comes into contact with the movable plug when the first and second screws are screwed together, and the movable plug is pressed and fixed toward the separating agent accommodated in the hollow tube. . Therefore, the separating agent can be easily pressed and the movable stopper can be fixed.

  In the present invention, of the first and second seals, the seal provided near the center of the hollow tube is fitted to the outer peripheral surface of the movable plug and gradually moves from the center to the end of the hollow tube. A first inner seal that forms a tapered portion with a reduced cross-sectional area, and a tapered portion that is fitted to the inner peripheral wall of the hollow tube and gradually increases in cross-sectional area from the center to the end of the hollow tube. The first outer seal, which is provided near the end of the hollow tube, is fitted to the movable stopper and is directed from the center to the end of the hollow tube. A second inner seal that forms a taper portion that gradually increases in cross-sectional area, and a taper portion that is fitted to the inner peripheral wall of the hollow tube and gradually decreases in cross-sectional area from the center to the end of the hollow tube. An inner seal and an outer seal in the first and second. It is preferable to contact a tapered portions. According to such a configuration, it is possible to further improve the sealing performance by the seal.

In the present invention, the supercritical fluid is carbon dioxide, the first seal is a seal made of polytetrafluoroethylene (PTFE), and the second seal is a seal made of polyvinylidene fluoride (PVDF). Preferably there is. Such a configuration is preferable from the viewpoint of preventing leakage of the mobile phase when carbon dioxide is used as the supercritical fluid.

  According to the present invention, the movable plug that is movable in the hollow tube along the axial direction of the hollow tube for containing the separating agent is used, and further, the gap between the movable plug and the inner peripheral wall of the hollow tube is closed. Since the seal prevents separation agent and solvent leakage and supercritical fluid leakage, the leakage of the mobile phase from the column is prevented in supercritical fluid chromatography, and the particulate separation agent is packed in the column. The state can be kept normal.

  Further, according to the present invention, when a fixing means that is threadably engaged with the first screw provided on the outer peripheral surface of the hollow tube and is movable along the axial direction of the hollow tube is used, In addition, the movable stopper can be easily fixed, which is more effective from the viewpoint of easily maintaining the charged state of the particulate separating agent in a normal state.

  According to the invention, when the first and second seals are composed of an inner seal and an outer seal that form tapered portions inclined in a predetermined direction and the tapered portions are in close contact with each other, the sealing performance of the seal is further increased. This is more effective from the viewpoint of preventing leakage of the mobile phase from the column.

  According to the present invention, when a seal made of polyvinylidene fluoride is used for the supercritical fluid of carbon dioxide, it is more effective from the viewpoint of preventing leakage of the supercritical fluid from the column.

  The column of the present invention is used for supercritical fluid chromatography using a fluid containing a supercritical fluid and a solvent as a mobile phase.

  The supercritical fluid is a substance that is in one or both of a state of a critical pressure or higher and a critical temperature or higher (that is, a supercritical state). Examples of substances used as the supercritical fluid include carbon dioxide, ammonia, sulfur dioxide, hydrogen halide, nitrous oxide, hydrogen sulfide, methane, ethane, propane, butane, ethylene, propylene, halogenated hydrocarbons, water, and the like. Can be mentioned.

  As the solvent, one kind or two or more kinds are selected from various known solvents according to the kind of the target substance, the kind of the supercritical fluid, and the like. Examples of the solvent include methanol, lower alcohols such as ethanol and 2-propanol, ketones such as acetone, acetonitrile, ethyl acetate, and water.

  The supercritical fluid chromatography is not particularly limited as long as it uses a fluid containing the supercritical fluid and the solvent as a mobile phase. The supercritical fluid chromatography may be used for analysis or for preparative use. The column of the present invention is preferably applied to preparative supercritical fluid chromatography. Preparative supercritical fluid chromatography is not particularly limited as long as it includes a step of separating the mobile phase after passing through the column by a fraction collector according to the target substance separated by the column. .

  The column of the present invention includes a hollow tube that contains a separating agent for separating a target substance, and plugs provided at both ends of the hollow tube so that a fluid can flow without leaking the separating agent.

The hollow tube is not particularly limited as long as it has a volume and an inner diameter sufficient to accommodate the separating agent and has a wall thickness sufficient to withstand the high pressure to which the mobile phase is supplied. Stainless steel can usually be used as the material.

  The inner diameter and volume of the hollow tube vary depending on the use of the column, the type of separation agent, the target production quantity of the target substance, etc., but when the target substance is separated, for example, the inner diameter is 20 to 600 mm. Preferably, it is 30 to 450 mm, and the volume of the part that accommodates the separating agent is 0.078 to 77 L, more preferably 0.18 to 40 L. The wall thickness of the hollow tube varies depending on the type and pressure of the mobile phase, the column material and the column size, etc., but in the case of a stainless steel hollow tube for fractionating the target substance, For example, it is preferably 4 to 55 mm.

  The stopper is not particularly limited as long as it is a stopper that allows fluid to flow without leaking the separating agent. The plug can be composed of, for example, a plug main body, a fluid flow path that penetrates the plug main body along the axial direction of the hollow tube, and a filter provided at the separating agent side outlet of the fluid flow path.

  In the present invention, a movable stopper that is movable in the hollow tube along the axial direction of the hollow tube is used for at least one of the stoppers. When the movable plug is housed in the hollow tube, a fluid passage that passes through the movable plug along the axial direction of the hollow tube, and a gap between the movable plug and the inner peripheral wall of the hollow tube And a seal for closing.

  The movable stopper has a fluid flow path for circulating a mobile phase, and a known movable stopper used in high performance liquid chromatography, simulated moving bed chromatography, or the like can be used.

  The seal may be any seal that can prevent leakage of the separating agent contained in the hollow tube and the mobile phase, that is, the supercritical fluid and the solvent. As the seal, a seal formed of a material having an appropriate material can be used according to the type of supercritical fluid, the pressure of the mobile phase, the type of solvent, and the like. The type of seal material and the number of seals are not particularly limited. That is, the seal may be composed of one seal formed of one kind of material, may be composed of one seal formed of a plurality of kinds of materials, or formed of a plurality of kinds of materials. A plurality of seals may be used.

  More specifically, the seal can be composed of a first seal and a second seal provided along the axial direction of the hollow tube. The first seal is a seal that prevents leakage of the separating agent and the solvent, and the second seal is a seal that prevents leakage of the supercritical fluid.

  The first seal includes, for example, a ring-shaped packing for preventing leakage of a mobile phase from the movable stopper in a movable stopper column used in high performance liquid chromatography or simulated moving bed chromatography. A known seal can be used. As the material for the first seal, a material having low mechanical permeability and suitable mechanical properties for closing the gap is used. Examples of such a material include materials known as the packing such as polytetrafluoroethylene (PTFE).

The second seal is not particularly limited as long as it can prevent leakage of the supercritical fluid. Such a second seal is appropriately selected according to the seal material, the type of supercritical fluid, and the like. For example, when the supercritical fluid is carbon dioxide, the material of the second seal is a material that has low carbon dioxide permeability to the supercritical fluid and has appropriate mechanical properties to close the gap. It is done. An example of such a material is polyvinylidene fluoride (PVDF). Moreover, as a form of a 2nd seal, seals, such as a ring-shaped packing, are mentioned similarly to a 1st seal. The form of the first and second seals may be the same or different.

  When the movable plug is accommodated in the hollow tube, the first and second seals are fitted to the movable plug or fitted into the hollow tube. It is not limited, Any may be provided near the center part of a hollow tube, and may be provided near the edge part of a hollow tube. The number of the first and second seals is not particularly limited, and each of them may be a single seal or a plurality of seals.

  If each of the first and second seals is composed of a plurality of seals in an appropriate form, it is possible to improve the adhesion of the seals. In the present invention, such a seal can be adopted as the seal.

  For example, of the first and second seals, the seal provided near the center of the hollow tube is fitted to the outer peripheral surface of the movable stopper and gradually has a cross-sectional area from the center to the end of the hollow tube. A first inner seal that forms a taper portion that shrinks, and a first taper portion that is fitted to the inner peripheral wall of the hollow tube and gradually increases in cross-sectional area from the center to the end of the hollow tube Among the first and second seals, the seal provided near the end of the hollow tube is fitted to the movable stopper and gradually has a sectional area from the center to the end of the hollow tube. A second inner seal that forms a taper portion that expands, and a second inner seal that is fitted to the inner peripheral wall of the hollow tube and forms a taper portion that gradually decreases in cross-sectional area from the center to the end of the hollow tube And the inner and outer seals in the first and second are tapered. When forming the first and second seal to be in close contact with each other, it is possible to enhance the sealing performance by the seal further.

  That is, when the seal near the center of the hollow tube receives pressure from the center near the hollow tube, the taper of the first inner seal is biased toward the taper of the first outer seal. Thus, the adhesion between the inner seal and the outer seal at both tapered portions is enhanced. The seal near the end of the hollow tube is also changed by the force corresponding to the separation agent contained in the hollow tube and the pressure from the mobile phase, so that the tapered portion of the second inner seal becomes the tapered portion of the second outer seal. It is urged | biased toward, and the adhesiveness of the inner side seal | sticker and outer side seal | sticker in both taper parts increases.

  The column of the present invention further has a fixing means for fixing the movable stopper in a pressed state along the axial direction of the hollow tube.

  The fixing means is not particularly limited as long as it can be fixed in a state where the movable stopper is pressed along the axial direction of the hollow tube. Known means can be employed as the fixing means. Examples of the fixing means include a screw-type cap, a plurality of bolts for tightening the movable stopper toward the center of the hollow tube, an oil pump, a manual jack, and the like.

  In the present invention, it is preferable that the fixing means is the screw-type cap from the viewpoint of ease of pressing and fixing, simplification of the configuration, and the like. That is, the hollow tube further includes a first screw provided around the outer peripheral surface of the end portion of the hollow tube, and the fixing means includes a peripheral wall portion surrounding the outer peripheral surface of the hollow tube, and an inner wall of the peripheral wall portion. It is preferable to have a second screw provided on the peripheral wall and screwed into the first screw, and a flange portion extending from the inner peripheral wall of the peripheral wall portion toward the central axis of the peripheral wall portion.

  The flange portion only needs to extend toward the central axis of the hollow tube sufficiently to abut directly or indirectly on the movable stopper. As such a screw-type cap fixing means, a known screw-type cap used in a movable stopper column used in high performance liquid chromatography or simulated moving bed chromatography can be used.

The hollow tube and the fixing means may further have a structure suitable for pressing and fixing by the fixing means. As such a configuration, for example, from the outer peripheral surface of the hollow tube or the fixing means, a pair of parallel surfaces (nut portions) formed by sandwiching these central axes, or from the outer peripheral surface of the hollow tube or the fixing means Examples include a protruding fixing flange portion.

  Moreover, the column of this invention may have other components other than the component mentioned above in the range which does not impair the effect of this invention. Such other components include, for example, the filter described above and the mobile phase that is provided at the hollow tube side outlet of the fluid flow path and flows through the fluid flow path in the cross-sectional direction of the hollow tube. Examples include a dispersion plate to be dispersed.

  As the filter, a known filter used in a movable stopper column used in high performance liquid chromatography or simulated moving bed chromatography can be used. The dispersion plate may be, for example, a disc provided with a plurality of holes, a disc provided with a plurality of concentric grooves, a disc provided with a plurality of radial grooves, or the like.

  The column of the present invention contains a separating agent for separating the target substance. The separating agent is not particularly limited as long as it is a separating agent that can separate a target substance. Since the column of the present invention is a column for supercritical fluid chromatography, it is preferably used for separation of optical isomers that are difficult to separate by chromatography such as high performance liquid chromatography using a liquid as a mobile phase. . Accordingly, the type of separating agent is appropriately selected according to the type of target substance. For example, if the target substance is an optical isomer, examples of the separating agent include optically active polysaccharides and polysaccharide derivatives. It is done.

  The polysaccharide may be any of a synthetic polysaccharide, a natural polysaccharide, and a natural product-modified polysaccharide as long as it is an optically active polysaccharide. The polysaccharide is preferably a polysaccharide with a high regularity of the binding mode, and is preferably a chain polysaccharide. Examples of the polysaccharide include various polysaccharides, and cellulose and amylose are particularly preferable.

  The polysaccharide derivative is not particularly limited as long as it is a polysaccharide derivative that can be used for separation of optical isomers. As such a polysaccharide derivative, for example, a polysaccharide derivative containing the polysaccharide as a skeleton, wherein at least a part of the hydroxyl group and amino group of the polysaccharide is substituted with a functional group that acts on an optical isomer in a sample. Can be mentioned. Examples of the polysaccharide derivative include various polysaccharide derivatives, and at least one selected from an ester derivative of cellulose, a carbamate derivative of cellulose, an ester derivative of amylose, and a carbamate derivative of amylose is particularly preferable. Specific examples of the polysaccharide derivative include various polysaccharide derivatives described in International Publication No. 95/23125 pamphlet.

  The separating agent may be in the form of particles, or in the form of particles such as the form supported on the particulate carrier, or supported on an integral carrier accommodated in the hollow tube. It may be in the form of a molded product that is integrally accommodated in a hollow tube, such as a molded product or a molded product made of a separating agent. In the present invention, the separating agent is preferably in the form of particles from the viewpoint of maintaining a normal state of the separating agent without forming a cavity between the separating agent and the inner peripheral wall of the hollow tube.

  Among the separating agents in the particulate form, the separating agent in the form of particles is, for example, as described in Japanese Patent No. 2783819, using the polysaccharide or polysaccharide derivative as the separating agent as a solvent. Dissolve the resulting solution dropwise into an insoluble solvent that does not dissolve the separating agent such as water, preferably an insoluble solvent containing a dispersing agent such as an anionic surfactant, with stirring. It is possible to manufacture by.

Further, the separating agent in the form of the particles can form a porous body. Such a separating agent, for example, disperses bubbles or polygen in the solution of the separating agent, disperses the dispersed solution in the insoluble solvent, distills or replaces the solvent from the dispersed separating agent solution, A method of washing the resulting separating agent particles with a washing solvent that dissolves polygen, if necessary, may be mentioned. As the polygen, known particles such as a resin and a salt, which are more soluble in the washing solvent than the particles of the separating agent to be produced, are used.

  Of the particulate separation agents, a separation agent in a form supported on a particulate carrier can be produced, for example, by supporting the separation agent on a carrier by physical adsorption or chemical adsorption. The separation agent is supported on the carrier by immersing the carrier in a solution containing the separation agent and, if necessary, another compound, reacting with the other compound as necessary, and distilling off the solvent in the solution, It can be carried out by a method of replacing the solvent in the solution with another solvent.

  Examples of the carrier include porous organic carriers such as polystyrene, polyacrylamide, polyacrylate, and derivatives thereof; porous materials such as silica, alumina, magnesia, glass, kaolin, titanium oxide, silicate, and hydroxyapatite. Inorganic carrier; and the like.

  The separation agent in the form of a molded product integrally accommodated in the hollow tube is prepared by, for example, storing the solution of the separation agent in a container having an appropriate shape in the production of the separation agent in the particulate form described above, and then adding the solvent. It can be produced by distilling or substituting to form the separating agent in the form of a molded product, or by supporting the separating agent on an integral carrier in the manner described above.

  Hereinafter, embodiments of the present invention will be described more specifically. FIG. 1 shows a column according to an embodiment of the present invention.

  As shown in FIG. 1, the column of the present embodiment includes a hollow tube 1, a fixed plug 2 provided on one end side of the hollow tube 1, and a movable plug 3 provided on the other end side of the hollow tube 1. And a fixing means 4 for fixing the movable plug 3 while being pressed along the axial direction of the hollow tube 1.

  The hollow tube 1 is a stainless steel tube, and screws corresponding to the first screw are provided on the outer peripheral surfaces of both ends. The hollow tube 1 has, for example, an inner diameter of 30 mm, a wall thickness (excluding an end portion where a screw is formed) of 6 mm, and a length of 300 mm. The outer peripheral surface excluding the surface and the screw is finished by # 400 buffing. A pair of parallel surfaces are formed on a part of the outer peripheral surface of the hollow tube 1.

  The fixing plug 2 includes a female screw cap 2a that is screwed into a screw provided on one end of the outer peripheral surface of the hollow tube 1, a liquid collecting plate 2b that is accommodated in the female screw cap 2a, and a female screw cap 2a. And a filter 2c provided on the liquid collecting plate 2b so as to face the inside of the hollow tube 1.

  The female screw cap 2 a includes a cylindrical fixing peripheral wall portion, a fixing flange portion extending from one end edge of the inner peripheral wall of the fixing peripheral wall portion toward the central axis of the fixing peripheral wall portion, and an outer periphery of the hollow tube 1. A screw provided on the inner peripheral surface of the fixing peripheral wall portion so as to be screwed to a screw provided on one end of the surface, and a fluid flow passing through the fixing flange portion along the central axis of the fixing peripheral wall portion. It is comprised from the path | route 2d.

  The liquid collecting plate 2b has, for example, a disk-like shape having a through hole at the center, and a plurality of concentric grooves and a plurality of radial grooves extending from the through holes provided on the surface on the filter 2c side. It is a member.

As the filter 2c, a filter having an appropriate opening is used according to the particle size of the separating agent accommodated in the hollow tube 1. The filter 2 c is formed in a disk shape having a diameter slightly larger than the outer diameter of one end of the hollow tube 1.

  In the fluid flow path 2d, a screw is provided on the distal end side of the female screw cap 2a so that a pipe through which a mobile phase flows can be connected. Further, the outer peripheral surface of the fixing flange portion of the female screw cap 2a is a nut portion 2e in which three parallel pairs of nuts are formed so that the cross-sectional shape is a hexagon.

  The movable plug 3 includes a movable plug main body 3a accommodated in the hollow tube 1, a fluid flow path 3b passing through the movable plug main body 3a along the central axis of the movable plug main body 3a, and the movable plug main body 3a being hollow. Seals 3c to 3f for closing gaps between the movable plug body 3a and the inner peripheral surface of the hollow tube 1 when accommodated in the tube 1, and an end surface of the movable plug body 3a facing the inside of the hollow tube 1 The dispersion plate 3g to be arranged and the regulating member 3h for suppressing the seal from the fixing means 4 side of the movable plug main body 3a are provided.

  The movable plug body 3a is formed in a shape in which a large-diameter portion and a small-diameter portion that are two cylinders having different diameters are connected. The large-diameter portion is formed with a diameter smaller than the inner diameter of the hollow tube 1, and the small-diameter portion is formed with a smaller diameter than the large-diameter portion.

  In the fluid flow path 3b, a screw is provided on the distal end side (the end side of the small diameter portion) of the movable plug body 3a so that a pipe through which the mobile phase flows can be connected.

  The seal 3c is a seal that is fitted on the movable plug main body 3a and forms a tapered portion in which the cross-sectional area gradually decreases from the center portion to the end portion of the hollow tube 1. The seal 3d is a seal that is fitted into the hollow tube 1 and forms a tapered portion in which the cross-sectional area gradually increases from the center portion to the end portion of the hollow tube 1. The tapered portions of the seal 3c and the seal 3d are in close contact with each other.

  The seal 3 e is a seal that is fitted in the hollow tube 1 and forms a tapered portion in which the cross-sectional area gradually decreases from the center portion to the end portion of the hollow tube 1. The seal 3f is a seal that is fitted on the movable plug main body 3a and forms a tapered portion in which the cross-sectional area gradually increases from the center portion to the end portion of the hollow tube 1. The tapered portions of the seal 3e and the seal 3f are in close contact with each other.

  The seal 3c and the seal 3d are made of polytetrafluoroethylene (PTFE), and the seal 3e and the seal 3f are made of polyvinylidene fluoride (PVDF). Note that the seal 3d and the seal 3e are integrally configured through, for example, a metal ring.

  The dispersion plate 3g is a disk-shaped member having, for example, through holes formed at a plurality of locations and a plurality of radial grooves extending radially from the center toward the through holes.

  The regulating member 3h is a donut-shaped member in which a through-hole having a small outer diameter is provided in the center of a disk-shaped member having an outer diameter approximately equal to the inner diameter of the hollow tube 1. The restricting member 3h is provided on the end face of the large-diameter portion opposite to the dispersion plate 3g, and is fixed to the movable plug main body 3a with a screw. The restricting member 3h is fixed to the movable stopper main body 3a so as to contact the seal 3f.

The fixing means 4 is configured in the same manner as the female screw cap 2a except that it has a through hole into which the small-diameter portion is fitted at the center of the flange portion. That is, the fixing means 4 is screwed into a peripheral wall portion 4a surrounding the outer peripheral surface of the hollow tube 1 and a screw provided at an end portion of the outer peripheral surface of the hollow tube 1 provided on the inner peripheral wall of the peripheral wall portion 4a. A screw 4b corresponding to the second screw and a flange portion 4c extending from one end edge of the inner peripheral wall of the peripheral wall portion 4a toward the central axis of the peripheral wall portion 4a. The flange portion 4c has a through hole in the center where the small diameter portion is fitted. The outer peripheral surface of the flange portion 4c is also formed in a nut shape like the female screw cap 2a.

Next, filling of the separation agent into the column according to the present embodiment will be described.
First, the separation agent is supplied to the hollow tube 1 in a state where the fixing tube 2 is provided in the hollow tube 1. As the separating agent, for example, particles comprising at least one polysaccharide derivative selected from cellulose ester derivatives, cellulose carbamate derivatives, amylose ester derivatives, and amylose carbamate derivatives, or the polysaccharide derivative is supported on particulate silica. The resulting particles are used. The separating agent is usually supplied in a slurry state.

  When the separation agent is supplied to the hollow tube 1, the separation agent is sucked from the fluid flow path 2 d using, for example, a vacuum pump so that the hollow tube 1 is filled tightly. Instead of suction from the fluid flow path 2d, or in addition, pressurization may be performed from the other end of the hollow tube 1 using, for example, a compressor, a pump, or a jack.

  When a predetermined amount of the separating agent is densely filled in the hollow tube 1, the movable plug 3 is installed from the other end side of the hollow tube 1. The movable plug 3 accommodates, for example, a movable plug main body 3a having a dispersion plate 3g and a seal 3c in the hollow tube 1 from the other end side of the hollow tube 1, and is provided with seals 3d and 3e and a seal 3f. It is installed by fixing the regulating member 3g to the movable plug body 3a.

  When the movable plug 3 is installed, the movable plug 3 is pressed along the axial direction of the hollow tube 1 by screwing the fixing means 4 to the other end side of the hollow tube 1. When the movable stopper 3 is appropriately pressed, the fastening of the fixing means 4 is stopped. Thus, the movable stopper 3 is pressed along the axial direction of the hollow tube 1 and is fixed in that state.

  The end point of pressing of the fixing means 4 may be determined by the supply amount of the separating agent and the position of the movable stopper 3 or the fixing means 4 with respect to the hollow tube 1, and the force required for tightening the fixing means 4 is a predetermined value. It is also good when it becomes.

  As shown in FIG. 1, the movable plug 3 is accommodated in at least the hollow tube 1 until the seal 3 f of the movable plug 3 is completely accommodated in the hollow tube 1. When the movable stopper 3 exists at this position, the height of the packed bed of the separating agent, that is, the distance from one end of the hollow tube 1 to the surface of the dispersion plate 3g becomes the maximum (for example, 250 mm).

Next, the usage form of the column of this Embodiment is demonstrated.
The column packed with the separating agent is installed in a supercritical fluid chromatography separation device. This supercritical fluid chromatography separation device includes, for example, a means for generating a mobile phase containing carbon dioxide and a solvent of a supercritical fluid, a fraction collector for separating the mobile phase according to the detection result of the detector, and a fraction collector And a means for recovering and reusing the carbon dioxide gas that had been formed from the mobile phase and formed the supercritical fluid. In the supercritical fluid chromatography separation device, the column of the present embodiment is installed so that the mobile phase flows from the fluid flow path 3b to the fluid flow path 2d. That is, the fluid channel 3b is connected to the upstream mobile phase channel, and the fluid channel 2d is connected to the downstream mobile phase channel following the detector.

  The mobile phase supplied to the fluid flow path 3b is evenly dispersed in the cross-sectional direction of the hollow tube 1 by the dispersion plate 3g. Therefore, the mobile phase flows through the hollow tube 1 at a uniform moving speed in the cross section of the hollow tube 1. The mobile phase that has circulated through the hollow tube 1 passes through the filter 2c. The filter 2c prevents the separation agent from flowing out. The mobile phase that has passed through the filter 2c is efficiently and evenly collected in the fluid flow path 2d by the liquid collecting plate 2b and discharged from the fluid flow path 2d.

  When the mobile phase is supplied into the hollow tube 1, the internal pressure of the hollow tube 1 is increased to the pressure of the mobile phase. At this time, the seal 3 c is pressed toward the other end of the hollow tube 1. When the seal 3c is pressed toward the other end of the hollow tube 1, the taper portion of the seal 3c is pressed toward the taper portion of the seal 3d, and the adhesion between the taper portion of the seal 3c and the taper portion of the seal 3 is increased. Is further increased. Thus, leakage of the solvent and the separating agent in the mobile phase is prevented.

  On the other hand, the seal 3f is pressed from the regulating member 3h by tightening with a screw when the regulating member 3h is fixed to the movable plug body 3a. Further, when the seal 3c is pressed toward the other end of the hollow tube 1, the seal 3f is further pressed from the regulating member 3h with a repulsive force corresponding thereto. Therefore, the taper portion of the seal 3f is pressed toward the taper portion of the seal 3e, and the adhesion between the taper portion of the seal 3e and the taper portion of the seal 3f is further enhanced. This prevents leakage of carbon dioxide supercritical fluid in the mobile phase.

  If the mobile phase is continuously supplied to the column of the present embodiment, the separating agent accommodated in the hollow tube 1 may be consumed and a cavity may be generated in the hollow tube 1. The generation of such a cavity can be detected, for example, by detection with a detector in the supercritical fluid chromatography separation apparatus or by periodically measuring the tightening force of the fixing means 4.

  When such a cavity is generated, the fixing means 4 is further tightened, the movable plug 3 is further pressed along the axial direction of the hollow tube 1, and the movable plug 3 is moved along the axial direction of the hollow tube 1. Further, the movable stopper 3 is appropriately pressed so that the cavity is filled. By repeating such position adjustment of the movable stopper 3, a normal filling state of the separating agent is maintained.

  In FIG. 2, the column of this Embodiment when the movable stopper 3 moves most by the press of the fixing means 4 is shown. When the movable plug 3 is completely accommodated in the hollow tube 1 and the movable plug 3 is present at this position, the height of the packed bed of the separating agent is minimized (for example, 245 mm).

  In the present embodiment, the configuration including the fixed plug 2 and the movable plug 3 is shown, but a movable plug may be provided in place of the fixed plug 2. According to such a configuration, it is possible to further press and fix the separating agent by a movable stopper in place of the fixed stopper 2, and it is possible to keep the separating agent filling state normal for a longer (almost twice) period. . In this case, it is preferable to provide a movable plug in place of the fixed plug 2 with a filter and a liquid collecting plate for preventing the separation agent from flowing out, as with the fixed plug 2.

  In this embodiment, when the column is installed in the supercritical fluid chromatography separation device, the column is installed so that the mobile phase flows from the fluid flow path 3b to the fluid flow path 2d. A column may be installed so that the mobile phase flows from the channel 2d to the fluid channel 3b. In this case, the movable stopper 3 is preferably provided with the liquid collecting plate instead of the dispersion plate, and further provided with a filter for preventing the separation agent from flowing out.

  According to this embodiment, when the hollow plug 1, the fixed plug 2, the movable plug 3, and the fixing means 4 are provided, and the movable plug main body 3a in the movable plug 3 is accommodated in the hollow tube 1. Since the seals 3c and 3d made of PTFE and the seals 3e and 3f made of PVDF are used for sealing the gap between the movable plug body 3a and the hollow tube 1, the supercritical fluid of carbon dioxide and the solvent In the supercritical fluid chromatography using the mobile phase containing the mobile phase, leakage of the mobile phase can be prevented and the packed state of the particulate separation agent can be kept normal. In addition, although the separation agent is usually filled at a pressure higher than that at the time of separation of the target substance, high hermeticity can be realized even under such a high pressure condition.

According to the present embodiment, a screw is formed on the outer peripheral surface of the end of the hollow tube 1, and the fixing means 4 includes
Since the fixing means including the peripheral wall portion 4a, the screw 4b, and the flange portion 4c is used, even when a cavity is generated in the filled separating agent layer, the fixing means 4 is further screwed to It is easy to keep the filling state normal.

  According to the present embodiment, the seal 3c has a tapered portion that is fitted on the outer peripheral surface of the large-diameter portion of the movable plug main body 3a and whose cross-sectional area gradually decreases from the center portion to the end portion of the hollow tube 1. The seal 3d is a seal that is fitted into the inner peripheral wall of the hollow tube 1 and forms a tapered portion that gradually increases in cross-sectional area from the center to the end of the hollow tube. 3e is a seal that is fitted into the inner peripheral wall of the hollow tube 1 to form a tapered portion that gradually decreases in cross-sectional area from the center to the end of the hollow tube, and the seal 3f is a movable plug body 3a. Is a seal that is externally fitted to the outer peripheral surface of the large-diameter portion and forms a taper portion in which the cross-sectional area gradually increases from the center portion toward the end portion of the hollow tube 1, and the taper portions of the seals 3c and 3d are respectively Since the respective taper portions of the seals 3e and 3f are in close contact with each other, the sheet during pressurization is Adhesion between 3c and 3d, and the adhesion between the seal 3e and 3f more can be further enhanced, it is more effective in terms of preventing leakage of the mobile phase.

  According to the present embodiment, since the dispersion plate 3 g is used, the mobile phase supplied from the fluid flow path 3 b spreads uniformly in the cross-sectional direction of the hollow tube 1. Therefore, regardless of the inner diameter of the hollow tube 1, the mobile phase can be passed uniformly through the separation agent layer accommodated in the hollow tube 1.

  According to the present embodiment, since the liquid collection plate 2b is used, the mobile phase that has passed through the separation agent layer can be efficiently collected in the fluid flow path 2d on one end side of the hollow tube 1. Therefore, the mobile phase can be collected from the hollow tube 1 having a large inner diameter to the flow path 2d of the fluid having a small inner diameter while suppressing disturbance of the flow of the mobile phase.

  According to the present embodiment, since the filter 2c is used, the separation agent accommodated in the hollow tube 1 can be prevented from flowing out of the hollow tube 1.

  According to the present embodiment, a pair of parallel surfaces are formed on the outer peripheral surface of the hollow tube 1, and the outer peripheral surfaces of the flange portions of the fixed plug 2 and the movable plug 3 are formed in a nut shape. 2 and the movable stopper 3 can be easily screwed in using an instrument such as a monkey wrench.

  According to this embodiment, since the hollow tube 1 has an inner diameter of 30 mm, a large amount of sample solution can be passed through the separation agent layer, and manufactured by fractionating the target substance in the sample solution. It is suitable for doing.

  According to the present embodiment, since the thickness of the hollow tube 1 is 6 mm, it can withstand the supply of a high-pressure mobile phase.

  According to this embodiment, since the separating agent is a particle having at least a polysaccharide derivative on the surface, when the target substance is an optical isomer, the target substance can be separated with high separation efficiency. Therefore, even when the filling height of the separating agent accommodated in the hollow tube 1 is relatively small as 245 to 250 mm, the target substance can be fractionated, and in the case of producing optical isomers by fractionation. A highly productive manufacturing apparatus can be configured, and the size of such a manufacturing apparatus can be reduced.

It is principal part sectional drawing which shows the column of one embodiment of this invention. It is principal part sectional drawing which shows the other state of the column shown in FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Hollow tube 2 Fixed plug 2a Female screw cap 2b Liquid collection board 2c Filter 2d, 3b Fluid flow path 2e Nut part 3 Movable plug 3a Movable plug main body 3c-3f Seal 3g Dispersion plate 3h Restriction member 4 Fixing means 4a Circumferential wall part 4b Screw 4c Flange

Claims (3)

A hollow tube for containing a separating agent for separating a target substance, and stoppers provided at both ends of the hollow tube for allowing a fluid to flow without leaking the separating agent, In a column used for supercritical fluid chromatography using a fluid containing a critical fluid and a solvent as a mobile phase,
At least one of the stoppers is a movable stopper that is movable in the hollow tube along the axial direction of the hollow tube,
The column further includes a fixing means for fixing the movable stopper in a pressed state along the axial direction of the hollow tube,
When the movable plug is accommodated in the hollow tube, a fluid flow path that penetrates the movable plug along the axial direction of the hollow tube, and the movable plug and the inner peripheral wall of the hollow tube And a seal that closes the gap between them,
The seal includes a first seal and a second seal provided side by side with respect to the axial direction of the hollow tube,
Said first seal is a seal for preventing leakage of the separating agent and the solvent, said second seal Ri Ah with a seal to prevent leakage of the supercritical fluid,
Of the first and second seals, the seal provided near the center of the hollow tube is fitted to the outer peripheral surface of the movable stopper and gradually cuts from the center to the end of the hollow tube. A first inner seal that forms a tapered portion with a reduced area, and a tapered portion that is fitted to the inner peripheral wall of the hollow tube and that gradually increases in cross-sectional area from the center to the end of the hollow tube. And a first outer seal that
Of the first and second seals, the seal provided near the end of the hollow tube is fitted to the movable stopper and gradually increases in cross-sectional area from the center to the end of the hollow tube. A second inner seal that forms a tapered portion that forms a taper portion that is fitted to the inner peripheral wall of the hollow tube and that gradually decreases in cross-sectional area from the center to the end of the hollow tube. And consists of an outer seal
The inner seal and the outer seal in the first and second are in close contact with each other between the tapered portions . The hollow tube further includes a first screw provided around the outer peripheral surface of the end portion of the hollow tube,
The fixing means includes a peripheral wall portion surrounding the outer peripheral surface of the hollow tube, a second screw provided on the inner peripheral wall of the peripheral wall portion and screwed into the first screw, and a peripheral wall from the inner peripheral wall of the peripheral wall portion. The column according to claim 1, further comprising a flange portion extending toward a central axis of the portion.   The supercritical fluid is carbon dioxide, the first seal is a seal made of polytetrafluoroethylene (PTFE), and the second seal is a seal made of polyvinylidene fluoride (PVDF). The column according to claim 1.

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