JPS629270A - Distribution column cartridge to be used for centrifugal liquid-liquid partition chromatographic device - Google Patents

Abstract

PURPOSE:To improve distribution performance by laminating distribution plates constituted by successively connecting the respective ends, on one side, of distribution pipes to the opposite ends of the adjacent distribution pipes by liquid connecting passages, further laminating sealing plates to both distribution plates on the outermost layer and opening the extreme ends of the liquid connecting passages of both distribution plates on the outermost layer to the side faces of the sealing plates in the same direction through the sealing plates. CONSTITUTION:Each distribution plate 1 is a rectangular plate-shaped body and is formed with plural pieces of groove-shaped distribution pipes 2 juxtaposed perpendicularly to the longitudinal direction thereof an equal intervals. The respective ends 4, on one side, of the pipes 2 are successively connected to the opposite ends 5 of the adjacent distribution pipes by the liquid connecting passages 3. The distribution pipe 2F at the extreme end is opened through the plate 1 to the bottom surface thereof by the liquid passage 3F at the extreme end. A stainkess steel is preferable for the material of the plate 1 in the case of metal and ethylenetrifluoride is preferable for said material in the case of plastics. The distribution performance is improved by using such distribution pipe column cartridge.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a distribution tube column cartridge used in a centrifugal liquid-liquid distribution chromatography apparatus.

A centrifugal liquid-liquid distribution chromatography device using the distribution tube column cartridge of the present invention is a device intended for large-scale fractionation and purification of physiologically active natural organic compounds used as pharmaceuticals, agricultural chemicals, cosmetics, foods, and feed additives. be.

(Prior art) Substances used for this purpose are extracted and purified from biological materials such as microbial culture media, microbial cells, animal and plant tissues, but in order to obtain the purified target substance, Conventionally, normal phase or reversed phase partition chromatography is often used. Partition chromatography usually uses silica gel, alumina gel, celite, diatomaceous earth, porous polystyrene resin, etc. as a carrier, and either physically coats the surface with a liquid phase or uses a column filled with chemically bonded particles. will be carried out. However, among the components extracted from biological materials, in addition to the target physiologically active substances, there are large amounts of pigments,
Due to the presence of impurities, the used carrier becomes irreversibly contaminated and must be frequently replaced and discarded. In addition, if the target physiologically active substance is strongly retained in the liquid phase (stationary phase) on the carrier surface or adsorbed to the carrier surface, the target substance will not be recovered from the column, and the conditions that allow it to elute from the column may be Under these conditions, physiologically active substances may be deactivated or denatured. The stationary phase adsorbed or bonded to the surface of a solid support is a very thin molecular layer and has an extremely small sample holding capacity. In order to fractionate and purify a large amount of samples, it is necessary to use a large diameter column filled with a large amount of carrier, which is not an advantageous method from both a technical and economic standpoint.

The difference in the partition coefficient of a substance between the two phases when one phase is the stationary phase and the other phase is the mobile phase in a mixture of two or more solvents that are not freely miscible with each other and separate into two phases. The method of separating and purifying each component from a mixed substance by multi-stage partition equilibrium using chromatography is known as countercurrent partition chromatography. Countercurrent partition chromatography does not use a solid support as a separation and purification medium, so it is easy to collect samples and clean the equipment, and separation is performed under mild conditions.
It has the advantage of stably recovering 100% of each component in the sample and having a large sample holding capacity, making it suitable for large-scale separation and purification. Flave countercurrent distribution devices and droplet countercurrent distribution devices are known as devices used for this purpose, but these have drawbacks such as complicated handling and operation and long time required for separation and purification. be.

The inventor of the present application previously developed countercurrent partition chromatography in a short time with high precision, and as a method suitable for separating and purifying a large amount of samples, one of the two-phase separated liquids is held as a stationary phase by centrifugal force. At the same time, he invented a centrifugal liquid-liquid distribution chromatography device in which the sample injected into the mobile phase was continuously collected in fractions by passing the other phase through the stationary phase continuously, and applied for a patent. (Special application 1986-1698
No. 00).

The principle of this device is shown in FIG. A stationary phase liquid 3 is held by centrifugal force in a large number of distribution pipes 1 arranged parallel to the direction of centrifugal acceleration and interconnected by thin tubes 2. A mobile liquid phase 4 is introduced from one end of the arranged distribution pipes through the thin tubes 2, and passes through the distribution pipe 1 in the form of fine droplets under the buoyancy generated by the action of centrifugal force. While the mobile phase liquid 4 continuously passes through the sequentially arranged distribution pipes 1,
Each component of the sample introduced into the mobile phase liquid 4 is
Multi-stage distribution is repeated between the liquid and the stationary phase liquid 3, and the liquid is separated according to the difference in distribution coefficient, flows out continuously from the other end of the arranged distribution pipe 1, and is collected as a fraction. The figure shows the liquid feeding direction when a liquid phase with a higher specific gravity of the two-phase separated liquid is used as a stationary phase and a liquid phase with a lower specific gravity is used as a mobile phase.

This apparatus, that is, a centrifugal liquid-liquid distribution chromatography apparatus using the distribution tube column cartridge of the present invention will be explained with reference to FIG.

In FIG. 2, one of the upper and lower phases of the two-phase separated distribution liquid is filled in advance as a stationary phase liquid into column cartridges 7 arranged radially around the rotor 8, and then the rotor 8 is rotated. While rotating the rotor 8, the other liquid is transferred from the solution tank 1 as a mobile phase liquid, and is sent through the upper recirculation liquid joint 5 from the rotating shaft 6 to the column cartridge 7. At the same time, a sample solution is injected from the sample injector 3. The mobile liquid phase is mixed with the stationary phase liquid in the column cartridge 7 and separated by centrifugal acceleration many times, during which components with different distribution coefficients in the sample liquid are separated. Each separated sample component passes through the center of the rotating shaft 6 and flows out through the lower rotating joint 5 together with the mobile phase liquid. The sample components in the mobile phase liquid that continuously flows out are detected by the ultraviolet/ray absorption monitor 14 (recorded by the recorder 15, and at the same time are fractionated in fixed amounts by the fraction collector 16. Collect both parts of the desired substance from.

(Problems to be Solved by the Invention) An object of the present invention is to provide a distribution tube column cartridge for use in the above-mentioned centrifugal liquid-liquid distribution chromatography apparatus, which has improved distribution performance compared to conventional ones, and which is suitable for mass production. An object of the present invention is to provide a distribution tube column cartridge that can be manufactured more easily.

(Means for Solving the Problems) As a result of intensive research, the inventor of the present application found that the rectangular plate-shaped body of the present invention has a plurality of pieces arranged in a single row or in multiple rows in parallel at equal intervals at right angles to the longitudinal direction of the rectangular plate-shaped body of the present invention. Laminated distribution plates each having a groove-shaped distribution pipe and connecting one end and the opposite end of each of the distribution pipes sequentially with a liquid passage, and further laminated with a seal plate on both distribution plates in the outermost layer. ,
The liquid passages at each end of each distribution plate are connected to the liquid passages at the ends of the adjacent distribution plate, and the ends of the liquid passages of both distribution plates in the outermost layer pass through the seal plate and are connected to the liquid passages at the ends of the adjacent distribution plates. Invented a distribution tube column cartridge for use in a centrifugal liquid-liquid distribution chromatography device, which is characterized by openings in the same direction, and furthermore, a plurality of column cartridges installed radially are tightly integrated and connected alternately. This objective was achieved by inventing a distribution tube column car ridge, which is characterized by forming an annular disk containing a large number of distribution tubes and liquid passages.

Next, the distribution tube column cartridge of the present invention will be specifically explained with reference to the drawings.

FIG. 3 is a schematic explanatory diagram of the distribution plate 1, which is one component of the distribution pipe column cartridge of the present invention, and FIG. 4 is a plan view and a side view thereof. In the figure, a distribution plate 1 is a rectangular plate-like body, and a plurality of groove-shaped distribution pipes 2 are formed in parallel at regular intervals perpendicular to the longitudinal direction of the distribution plate 1, and each of the distribution pipes 2 has one end thereof. 4 is sequentially connected to the opposite end 5 of the adjacent distribution pipe through a liquid passage 3, and the distribution pipe 2F at the final end penetrates the distribution plate 1 through a liquid passage 3' at the final end and opens on the lower surface thereof. ing. The material of the distribution plate 1 may be metal, plastic, or ceramic, and in the case of metal, SUS is preferable, and in the case of plastic, trifluoroethylene is preferable.

Figure 5 shows the steps to configure the distribution tube column cartridge.
It is an explanatory view showing the mode of lamination of the above-mentioned distribution plate. In the figure, the final end liquid passage 3' of the first distribution plate communicates with the liquid passage 31 to the first distribution pipe of the second distribution plate arranged in the opposite direction, and the final end passage 3' of the second distribution plate The liquid passage 3' communicates with the liquid passage 3 to the first distribution pipe of the third distribution plate arranged in the opposite direction thereto, and in this way, the liquid passage 3' connects to the final end of the last distribution plate. Several distribution plates are stacked one on top of the other such that the channels 3' are located at the ends of the same side of the channels 3' of the first distribution plate to the first distribution tube. The stacked body composed of a plurality of distribution plates constructed in this way is sealed and fixed by sandwiching the upper and lower surfaces between seal plates 7. At this time, the liquid passage 31 of the first distribution plate and the final end liquid passage 3F of the last distribution plate penetrate the upper and lower seal plates 7 and open in the same direction 8 and 9 on the side surface of the seal plate.

As a means for fixing the distribution plate stack with the seal plates, an appropriate number of holes may be made through the periphery of both seal plates and each distribution plate, and the holes may be tightened with bolts and nuts. A sheet 6 is usually inserted between the distribution plates and between the air supply plate and the seal plate, and Teflon is preferably used as the sheet.

Although a distribution pipe column cartridge formed by stacking distribution plates in which distribution pipes are arranged in a single row has been described above, the distribution pipes can also be arranged in multiple rows on the distribution plate. FIGS. 6 and 7 are explanatory diagrams showing a plan view, a side view, and a stacking mode when the distribution pipes are arranged in two rows. It is clear that even in the case of a multi-row arrangement, the liquid passage 31 of the first distribution plate and the final liquid passage 3' of the last distribution plate must open in the same direction on the side surface of the sealing plate.

Furthermore, since the distribution tube column cartridge of the present invention is arranged radially around the rotor as described above, a plurality of distribution tube column cartridges arranged radially are closely integrated and alternately connected. It can be formed into an annular disk containing a large number of distribution pipes and liquid passages.

8 and 9 are a schematic plan view and a vertical sectional view of a disc-stacked cartridge formed in an annular disc.

In the figure, a distribution pipe 1 is formed by carving a large number of fan-shaped moats in a disc made of resin (preferably fluorinated hydrocarbon resin), metal (SUS), or ceramics, and the distribution pipes are connected to each other on one side of the disc. Connected by a groove 2 carved into the top layer disk 1
The connection groove 3 connected to the first distribution pipe of 0 is connected to the upper liquid feeding joint 7 attached to the upper holding plate 5 through the seal 9, and the connection groove 4 connected to the last distribution pipe is carved on the back side of the disk. , connects to the connecting groove connecting to the first branch pipe of the second disc through the seal, and connects the stacked discs in the same way sequentially, and the connecting groove connecting to the last dividing pipe of the lowest disc 10 is sealed. It is connected to a lower liquid feeding joint 8 attached to the lower holding plate 6 through the opening. FIG. 10 shows a configuration in which a disc-stacked distribution tube column cartridge is assembled into a rotor. In the figure, 11 is a rotation liquid joint, 12 is a rotation joint, 13 is a connection tube, 14 is a connection tube, 15 is a disk light plate, 1
6 is a bearing, and 17 is a pulley.

[Brief explanation of the drawing]

FIG. 1 is an explanatory diagram of the principle of a centrifugal liquid-liquid distribution chromatography apparatus using the distribution tube column cartridge of the present invention. FIG. 2 is an explanatory diagram of the centrifugal liquid-liquid distribution chromatography apparatus. FIG. 3 is a schematic explanatory diagram of a distribution plate which is a component of the distribution pipe column cartridge of the present invention. FIG. 4 is a plan view and a side view of the distribution plate. FIG. 5 shows an embodiment of the stacking of distribution plates for constructing the distribution tube column cartridge of the present invention. FIG. 6 is a plan view and a side view when the distribution pipes are arranged in two rows. FIG. 7 is a diagram illustrating the stacking mode when the distribution pipes are arranged in two rows. FIGS. 8 and 9 are a schematic plan view and a vertical sectional view of a disc-stacked type distribution tube column cartridge. FIG. 10 is a schematic diagram of a structure in which a disc-stacked distribution pipe column cartridge is assembled into a rotor. -X/Kokoshu 2 Zuge 4wJ 纂5I! I Oro Zujo 7U! J tail a figure low U--L” ninini ni 70 2222 = mouth q figure

Claims (1)

[Scope of Claims] 1) Consisting of a cylindrical rotor with rotating liquid joints at both the upper and lower ends of the rotating shaft and a plurality of column cartridges installed radially along the periphery; The liquid joint is connected to any one column cartridge, and the column cartridge is sequentially connected to adjacent column cartridges, and the last column cartridge is connected to a rotary transfer liquid joint at the lower end of the rotating shaft. In a graphing device, a rectangular plate-like body has a plurality of groove-shaped distribution pipes arranged in a single row or in multiple rows at equal intervals perpendicular to the longitudinal direction of the plate-shaped body, and adjacent to one end of each of the distribution pipes. Distribution plates made by sequentially connecting the opposite ends of the distribution pipes with liquid passages are stacked, and a seal plate is further laminated on both distribution plates in the outermost layer, so that the liquid passages at each end of each distribution plate are adjacent to each other. It is characterized in that it is connected to a liquid passageway at the end of the distribution plate, and the extreme ends of the liquid passageways of both distribution plates in the outermost layer penetrate the sealing plate and are opened in the same direction on the side surface of the sealing plate. Distribution tube column cartridge used in centrifugal liquid-liquid distribution chromatography equipment. 2) The distribution pipe column cartridge of item 1, in which a sheet is interposed between the stacked distribution pipes and the distribution pipes, and between the distribution pipes and the seal plate. 3) The distribution pipe column cartridge according to item 1, wherein the rectangular plate-shaped body is made of metal, plastic, or ceramic, and the seal plate is made of metal. 4) It consists of a cylindrical rotor with rotating liquid joints at both the upper and lower ends of the rotating shaft, and a plurality of column cartridges installed radially along the periphery. In a centrifugal liquid-liquid distribution chromatography device, the column cartridges are connected to adjacent column cartridges in sequence, and the last column cartridge is connected to a rotating liquid joint at the lower end of a rotating shaft. A centrifugal liquid-liquid distribution chromatograph apparatus characterized in that a plurality of installed column cartridges are closely integrated to form an annular disk containing a large number of distribution pipes and liquid passages connected alternately. Distribution tube column cartridge used for.