JPH0438290Y2 - - Google Patents

Description

[Detailed description of the invention] [Industrial application field] This invention is suitable for analysis and separation and purification, especially quantitative and qualitative analysis of biological substances, natural organic compounds, etc., and purification of them from crude samples. This invention relates to a centrifugal countercurrent distribution chromatography device.

[Conventional technology]

When performing countercurrent partition chromatography continuously, mixing and separating the stationary phase liquid and mobile phase liquid under a centrifugal acceleration field can significantly shorten the separation and analysis time and eliminate the limitations imposed by the partition liquid system. It also disappears. Various centrifugal countercurrent distribution chromatograph devices have been devised based on this idea, but all of these devices are coiled separation tubes installed inside a centrifuge.
Since the separation is performed by a method, if you want to increase the amount of sample to be separated and analyzed, you have to increase the inner diameter of the separation tube. However, if the separation tube is made thicker, the distribution of substances between the mobile phase liquid and the stationary phase liquid will not be sufficient, and the spread of the separation peak due to diffusion will also increase.Therefore, the method using a coiled separation tube However, there was a limit to the amount of sample that could be separated.

Therefore, the same applicant as the present applicant devised a centrifugal countercurrent distribution chromatography device that solved the above-mentioned problems at once.
No.) has already been filed. The present invention is characterized by a cassette that includes one continuous separation tube and is detachably attached to the outer circumference of a rotating body. To explain this cassette with reference to FIGS. 8 to 11, this cassette 1 is composed of a main body 2 made of synthetic resin, and metal side plates 3 screwed to both sides of the main body 2 through sealing material 4. . The main body 2 is provided with two rows of large diameter holes 5a, 5b and small diameter holes 6a, 6b, respectively.
The large diameter holes 5a, 5b are inclined inwardly at equal angles, and the small diameter holes 6a, 6b are parallel to each other. The large-diameter holes 5a, 5b and the small-diameter holes 6a, 6b communicate with each other through communication grooves 7 alternately at their inner ends, and there is one continuous separation line from the inlet 8 to the outlet 9. A tube is formed. Furthermore, 10,
Reference numeral 11 denotes a tube attachment collector provided at the inlet 8 and the outlet 9.

As shown in FIG. 7, when installing, for example, two cassettes 1a and 1b on the rotating body 12 of the centrifugal countercurrent distribution chromatography device, first the insertion The cassettes 1a and 1b are inserted into the hole 13 at 180° intervals, and the cassettes 1a and 1b are communicated with each other through the connecting tube 14, and the inlet 8 of one cassette 1a is connected to the cassettes 1a and 1b.
and the outflow port 9 of the other cassette 1b are connected to the supply pipe and the outflow pipe via rotary joints (not shown) provided at the upper and lower ends of the rotating shaft 17, respectively. be done.

The above device has two types of holes 5, large and small, in the cassette 1.
A, 5b, 6a, 6b are provided in large numbers and their ends are connected alternately to form one continuous separation tube, so it is possible to increase the number of effective theoretical plates per unit separation tube length. , the stationary phase liquid is retained in the large-diameter holes 5a and 5b, and the volume of the stationary phase liquid can be increased and the volume ratio of the stationary phase liquid and the mobile phase liquid can be increased.As a result, many samples can be Can be separated in a short time. Also, cassette 1
If a and 1b are attached to and detached from the rotating body 12 as appropriate,
This provides an extremely significant effect in that the length of the separation tube can be easily changed.

[Problem that the invention attempts to solve]

By the way, by using the above-mentioned cassette, it is possible to obtain a centrifugal countercurrent distribution chromatography device which exhibits an epoch-making effect, but there are some problems when putting it into practical use. That is, the cassette 1 has a block-like main body 2 with a large number of holes 5a, 5b, 6a, 6.
However, it is difficult to precisely position the large number of holes 5a, 5b, 6a, and 6b and drill them to a predetermined diameter through machining, and the large diameter holes 5a, 5b are inclined inwardly at equal angles, and each hole 5a, 5b,
Since 6a and 6b must be communicated through the communication groove 7, extremely complicated and highly accurate machining is required. For this reason, this cassette 1 is very expensive and has a poor yield, which in turn becomes a factor in increasing costs.

This invention was devised in view of the above-mentioned conventional problems and to solve them.
This attempt was made to provide a centrifugal countercurrent distribution chromatography apparatus in which a cassette having multiple separation tubes can be manufactured by a relatively simple process.

[Means for Solving the Problems] In order to achieve the above-mentioned problems, this invention has a structure in which a large number of large-diameter holes and small-diameter holes are formed by laminating separation members of the same shape that are easy to process through a seal plate. The large-diameter holes and the small-diameter holes were alternately communicated at each end to form a rotating body having one continuous separation tube. That is, the centrifugal countercurrent distribution chromatography device according to this invention forms one continuous separation tube in the rotating body, and connects the external supply pipe and outflow via each rotary joint provided at the center of the rotating body. In an apparatus in which a tube and both ends of the separation tube are respectively connected and a sample is continuously supplied from the supply tube while rotating the rotating body to perform countercurrent distribution, a disk is provided with:
A plurality of elongated holes penetrating from one side to the other side are radially bored, and between a pair of adjacent elongated holes on one side of the disk, a radial direction of the one elongated hole is formed. A supply groove is formed in which the outer end and one end communicate with each other, and between the pair of adjacent elongated holes on the other side of the disk, the radially inner end and one end of the other elongated hole are formed. communicate with each other to form an outflow groove whose other end corresponds to the position of the other end of the supply groove, and between each elongated hole other than the pair of adjacent elongated holes on both sides or one side of the disc. A plurality of disk separators are formed by forming separation grooves each having a small width extending in the radial direction, and connecting each of these elongated holes and each separation groove alternately through a communication groove at each end. Configured with the necessary features. The plurality of disc separators and the plurality of seal plates each having a communication hole are arranged alternately so that each disc separator faces the same direction and the other end of each of the supply grooves is arranged alternately. and stacked and joined so that the other end of each of the outflow grooves and the communication hole of each seal plate communicate with each other, and
Seal plates in which communication holes are formed are interposed between the disc separators located at both ends of the laminate, so that the side plate in which the supply port is formed and the side plate in which the outflow port is formed are connected to the supply port and the side plate in which the outflow port is formed, respectively. The rotating body was constructed by joining and fixing the outlet, one end of the supply groove, and one end of the outflow groove so that they communicated with each other. Further, the opening surface of each of the supply grooves, both opening surfaces of each of the elongated holes, the opening surface of each of the communication grooves, the opening surface of each of the separation grooves, and the opening surface of each of the outflow grooves are connected to each of the seal plates. 1 leading from the supply port to the outflow port by each supply hole, each large diameter hole, each communication hole, each small diameter hole, each outflow hole, and each communication hole of each seal plate, which are each formed by being closed. The gist of the present invention is that the separation tube is made up of a series of books.

[Effect]

With the above configuration, a plurality of disk separators having a relatively simple shape and formed radially with long holes and separation grooves arranged alternately on disks can be stacked with a sealant interposed therebetween. So, the aforementioned special public service 1386-1986
A rotary body having the same function as the rotary body described in the above publication and having one continuous separation tube is constructed.

〔Example〕

Hereinafter, a preferred embodiment of this invention will be described.
This will be explained in detail with reference to the drawings.

As shown in FIG. 1, the rotating body 18 includes a plurality of (five in this embodiment) disk separation bodies 1 having the same shape.
9 are laminated with a seal plate 20 in between, and metal side plates 21 are joined to each side via the seal plate 20 and fixed by a threaded rod 22.

As shown in FIGS. 3 and 4, in the disc separator 19 described above, a mounting hole 19b into which the rotation shaft 23 is fitted is formed in the center of a disc 19a made of synthetic resin. A large number of long holes 19c are drilled radially from the long holes 19c, and separating grooves 19d having a width sufficiently smaller than the long holes 19c are formed on both sides of the disk 19a between the long holes 19c. The ends of the and separation grooves 19d are alternately connected to the communication groove 1.
9e to communicate with each other. As shown in Fig. 3, in some separation groove formation locations, there are supply separation grooves 1 on both sides with approximately half the length.
9f and an outflow separation groove 19g are formed, and a large number of insertion holes 31 for the threaded rod 22 are arranged along both ends of the elongated hole 19c and the separation groove 19d.

As shown in FIG. 2, this disk separator 19 is
The respective supply separation grooves 19f and outflow separation grooves 19g are positioned at the same position and stacked, and the communication holes 20' of each seal plate 20 and the supply ports 21' and outflow ports 21'' of both side plates 21 supply The elongated hole 19
A large diameter hole 24 in which c is closed by the seal plate 20 from both sides, and a small diameter hole 25 in which the separation groove 19d is closed by the seal plate 20 are connected to the communication groove 19e.
The communication hole 26 is closed by the seal plate 20.
As a result, the respective ends are alternately communicated with each other, and the outflow separation groove 19g and the supply separation groove 19f of each disk separator 19 are communicated with each other through the communication hole 20' of the seal plate 20, and the supply port 21' Outlet 2
A continuous separation tube extending to 1" is formed.
Collects 27 and 28 for attaching tubes are attached to the supply port 21' and the outlet 21'', respectively.

The operation of the embodiment apparatus having the above-described configuration will be described with reference to FIGS. 5 and 6.

First, all the holes 24, 25, and 26 are filled with stationary phase liquid A having a high specific gravity. Then, while rotating the rotating body 18 by a motor (not shown), a mobile phase liquid B with a small specific gravity is pumped through a supply pipe 32 shown in FIG. 5 by a pump (not shown), and the mobile phase liquid B The inlet 2 passes through the tube 29 connected via the rotary joint 33 of the rotary shaft 23.
1' to begin countercurrent distribution. That is, the mobile phase liquid B flowing from the inlet 21' into the supply separation groove 19f of the uppermost disk separator 19, that is, the small diameter hole 25, flows as shown in the double-dashed line in FIG. 3 while pushing the stationary phase liquid A. As shown, the water passes through the small diameter hole 25 and flows into the large diameter hole 24 from the end of the large diameter hole 24, that is, the end farthest from the rotating shaft 23 of the rotating body 18. At this time, a larger centrifugal force acts on the stationary phase liquid A, which has a higher specific gravity, than the mobile phase liquid B, which has a lower specific gravity, so the mobile phase liquid B becomes small particles and is centrifuged, as shown in Figure 6. The liquid moves to the opposite side to the direction in which the force is applied, that is, to the right in FIG. 6, and distribution separation is performed between the mobile phase liquid B and the stationary phase liquid A. At this time, since the large-diameter hole 24 is provided in the radial direction of the rotating body 18, the particles of the mobile phase liquid B pass through the large-diameter hole 24 without being biased to one side. And stationary phase liquid A
The mobile phase liquid B that has passed through the inside accumulates on the end face of the large-diameter hole 24, is collected at the inner end by the action of centrifugal force, and flows into the next small-diameter hole 25 via the communication hole 26. In this way, countercurrent distribution is repeated, and when a steady state is reached, the stationary phase liquid A is held in the large diameter hole 25, and the mobile phase liquid B is filled in the small diameter hole 25, and the total volume is filled as a whole. Stationary phase liquid A occupies 4/5. The mobile phase liquid B that has passed through the separation tube and has been fractionated and purified is transferred to the tube 30 and the rotary joint 34.
It flows out from the outflow pipe 35 through.

Note that this invention is not limited to the above description and drawings, and various modifications can be made without departing from the scope of the claims. For example, the separation groove may be formed only on one side of the disk. Good too.

〔effect〕

As explained above, in the centrifugal countercurrent distribution chromatography device according to this invention, a large number of long holes are arranged radially in the disk, and a small width separation groove is provided between each long hole. By stacking multiple disc separators with a seal plate in between, the ends of the large-diameter holes and the small-diameter holes are alternately connected with each other through communication grooves. By configuring the rotating body to have a single separation tube connected to it, it is possible to manufacture a rotating body that can separate many samples in a short time using an extremely simple molding process, which in turn leads to significant cost reductions. Connect.

[Brief explanation of the drawing]

1 to 6 show one embodiment of the centrifugal countercurrent distribution chromatography device according to the invention, in which FIG. 1 is a partially cutaway front view, FIG. 2 is a partially longitudinal sectional view, and FIG. 4 is a perspective view of a part of the disk separator, FIG. 5 is a schematic explanatory view, and FIG. 6 is an explanatory view showing the state of countercurrent distribution. Furthermore, FIG. 7 and subsequent figures show the conventional device, with FIG. 7 being an exploded perspective view of the entire device, FIG. 8 being a partially cutaway front view of the cassette, and FIG. 9 being a right side view of the cassette with the side plate removed. FIG. 10 and FIG. 11 are a sectional view taken along the - line and a sectional view taken along the - line of FIG. 9, respectively. 18... Rotating body, 19... Disc separation body, 19a
...Disk, 19c...Long hole, 19d, 19f, 1
9g...Separation groove, 19e...Communication groove, 20...Seal plate, 21...Side plate, 24...Large diameter hole, 25...
...Small diameter hole, 32... Supply pipe, 33, 34... Rotary joint, 35... Outflow pipe.

Claims (1)

[Scope of utility model registration request] One continuous separation pipe is formed in the rotating body 18, and the external supply pipe 32 and the outflow pipe 35 are connected to both ends of the separation pipe through rotary joints 33 and 34 provided at the center of the rotating body 18. and the supply pipe 3 while rotating the rotating body 18.
In a centrifugal countercurrent distribution chromatography device that performs countercurrent distribution by continuously supplying a sample from 2, a plurality of elongated holes 19c are formed in the disk 19a, penetrating from one side to the other side. In addition to drilling holes in a radial pattern, one side of the disk 19a,
A supply groove 19f is formed between a pair of adjacent elongated holes 19c, 19c, and the radially outer end of one of the elongated holes communicates with the other end.
Between the pair of adjacent elongated holes 19c, 19c, one end communicates with the radially inner end of the other elongated hole, and the position of the other end corresponds to the position of the other end of the supply groove 19f. 19g of outflow grooves were formed, and the disc 1
Separation grooves 19d having a small width extending in the radial direction are formed between each of the long holes 19c, 19c on both sides or one side of the long hole 9a other than the pair of adjacent long holes, and each of these long holes 19c and each separation groove are formed. 19d are alternately communicated with each other through communication grooves 19e at their respective ends, and each of the plurality of disk separators 19 is connected to a communication hole 20'.
A plurality of seal plates 20 formed with are arranged alternately so that each disk separator 19 is oriented in the same direction, and the other end of each of the supply grooves 19f and the other end of each of the outflow grooves 19g are arranged alternately. and the communication hole 2 of each seal plate 20
0' are laminated and joined so that they communicate with each other, and a sealing plate 20 in which a communication hole 20' is formed is interposed between each of the disk separation bodies 19, 19 located at both ends of the laminated body, The side plate 21 in which the supply port 21' is formed and the side plate 21 in which the outlet port 21'' is formed are connected to the supply port 21' and the outlet 21'', one end of the supply groove 19f, and one end of the outflow groove 19g, respectively. The rotary body 18 is constructed by joining and fixing them so that they communicate with each other, and the opening surface of each of the supply grooves 19f, each of the long holes 19c
, the opening surfaces of each of the communication grooves 19e, the opening surfaces of each of the separation grooves 19d, and each of the outflow grooves 1.
Each supply hole and each large diameter hole 2 are formed by closing the opening surface of 9g with each seal plate 20.
4. Each communication hole 26, each small diameter hole 25, each outflow hole, and the communication hole 20' of each seal plate 20 form one continuous separation pipe from the supply port 21' to the outflow port 21''. A centrifugal countercurrent distribution chromatography device characterized by comprising: