JPS617466A - Method and apparatus for filling of column in liquid chromatograph - Google Patents

Abstract

PURPOSE:To improve separation efficiency of a subjected material, by fitting under pressure a movable plug mounted on a column inside the column with a mechanical transmission mechanism and charging the column with a filling material under pressure. CONSTITUTION:After pouring concentration adjusted slurry filling material into a column 1 fixed from above onto a supporting bench 6, a movable plug 2 is fit and further, an electric motor 5 is driven to a constant speed by setting a travelling speed of the movable plug, allowing the plug 2 to drop through reduction gear 7, rotating shafts 4A, 4B. Thus, the poured slurry of the filling material is filled under a constant load and when the set load is found in excess of the set value in the neighborhood of the completed state of filling, the motor 5 is brought to a stand still to finish up the filling operation. As a reduction ratio of the reduction gear 7 is set in a wide range for the movable plug 2 of the column, the plug 2 is not subjected to any movement by fluid pressure at the time of analysis or blelding, etc., and consequently, the column can be used as a backed column even if the pressure is approximately 200kg/cm<2>G.

Description

DETAILED DESCRIPTION OF THE INVENTION 3.1 Technical Field The present patent relates to a method and apparatus for filling a column, which is a main part of a liquid chromatograph, with a packing material suitable for separating target component substances.

3.2 Prior art and its problems Conventional liquid chromatographs use columns with limited packing material and limited method as separation columns, mostly pre-packed commercially available punctured columns. However, in recent years, there has been a trend toward bulk separation of biochemical substances and pharmaceuticals based on liquid chromatography separation technology, and in order to do this more efficiently, we have developed packing materials that are optimal for separating the target substances. Liquid chromatographs equipped with packing devices that can increase separation efficiency by using columns packed in an appropriate manner have come into use.

However, the filling devices that have been used so far have the following problems, making them inconvenient for users.

(1) This method uses fluid pressure to pressurize the column containing the packing material from the outside to increase the packing density of the contents.In relation to the balance with the working pressure, the low pressure side should be filled with a movable stopper. The entire filling material is moved by a movable stopper and is filled under pressure against a fixed wall.

In this case, all the packed particles of the packing material move, causing friction between the particles and the column wall or between particles, which deteriorates the adsorption performance and extremely reduces the number of theoretical plates.

(2) For fluid pressure, a movable stopper is used on the low-pressure side when the column is used; however, in analysis or preparative separation, the sample or eluent is flowed under pressure, so force is applied to the movable stopper from within. Therefore, in the filling device using this method, it is necessary to constantly apply fluid pressure, which is the filling pressure, during use.

(3) In the situations described in F (+) and (2), the moving load of the movable stopper becomes uneven or unreasonable, making it impossible to fill with a constant load. Further, in order to prevent movement of the movable stopper due to fluid pressure, additional pressure must be applied from the outside [7]. Therefore, it is necessary to limit the working pressure to 20 kg/cm'G or less. However, 20Kg/cm'
Sufficient functionality cannot be achieved with a certain amount of pressure. That is, the packing density of the filler cannot be increased, and the number of theoretical plates cannot be improved either.

3.3 Purpose of the present invention The method and apparatus presented here solve these problems, and are suitable for use in liquid chromatographs, allowing even inexperienced people to easily fill the packing material, and also leading to an increase in the number of theoretical plates. A method and apparatus for column packing.

3.4 Structure of the present invention The basic feature of the present invention is that the packing material is pressurized by pressurizing a movable plug attached to the - side inlet of the column into the inside of the column using a mechanical conduction mechanism. A method for filling a column for liquid chromatography is provided, further comprising a movable stopper, a mechanical conduction mechanism for press-fitting the movable stopper into the inside of the column, and a pedestal for fixing the column. A packing device for a liquid chromatography column is provided.

The device provided by the present invention is equipped with a highly efficient filling mechanism using a movable plug system or a movable column system, and the movable plug part has a sealing part made of a material and shape that is chemical-resistant, wear-resistant, and preferably has a scraper structure, and has a pressure-resistant structure. 200Kg/cm'G
This structure has a high sealing performance. In addition, the movable plug part is designed to be removable for injecting the filler and maintaining the mechanism, and furthermore, it is possible to ensure a constant load during filling.

The present invention will now be described in detail with reference to the drawings. For convenience of explanation, a specific example will be shown, but of course the configuration of the present invention is not limited to this.

FIG. 1 is a schematic configuration diagram showing a packing material filling apparatus according to the present invention, in which a liquid chromatography column 1 is mounted and fixed on a column holder 8. In FIG. In this state, the movable stopper 2 has been removed and the movable stopper moving base 3 has risen to the top, and at this time, the limit switch is activated and electrical and mechanical considerations are taken to prevent it from rising any further. ing. The slurry-like packing material that has been adjusted once is injected into the column 1 from the top of the column 1, and the movable stopper 2 is attached.
Fixed to. Then, the movable stopper moving load is set and the electric motor 5
The movable stopper 2 and the movable stopper moving base 3 are rotated at a constant speed through the deceleration mechanism 7, and the movable stopper 2 and the movable stopper moving base 3 are rotated along the rotation shafts 4^, 4B in accordance with the rotation of the electric motor 5.
It rises or falls by rotating. The injected filler slurry is filled with a constant load. Near the completion of filling, when the preset load is exceeded, the electric motor 5 is driven out due to the excess of the limit load, and the filling is completed. The movable stopper 2 of the wooden column has a large reduction ratio of the reduction mechanism 7, so that the movable stopper 2 does not move due to fluid pressure during analysis or preparative use, so it weighs 200 kg.
It can be used in the same way as a backed column even at a pressure of about /cm'G.

When the filled packing material is used up, it is extracted from the bottom of the column 1, and when the filling is finished, the movable stopper 2 is removed from the movable stopper moving base 3 to prepare for the next filling.

Fig. 2 shows that in the apparatus explained in Fig. 1, the internal slurry is filled with a constant load or a constant load even though the movable stopper and the column (or the movable column and the fixed column) move relative to each other. This figure shows the configuration of the seal part and scraper to make this possible.

The seal structure adopted in the present invention has the problem that in the conventional method, particulate matter in the slurry gets between the O-ring and the column wall, deteriorating the sealing performance and shortening the life of the O-ring. On the other hand, it has a structure in which an O-ring and a scraper ring are combined, and two sets are arranged above and below with a spacer in between to ensure sealing performance when moving upward or downward.

In FIG. 2, when the movable stopper 2 descends and applies pressure to the screw in the column to uniformly fill the particulate matter, the liquid discharged due to the pressure applied to the slurry in the column is separated by the filter 16. , are discharged to the outside from the pipe 18 via the joint 17.

The 5f movable stopper tip 12 moves downward while applying pressure to the slurry, and the slurry moves between the column tube wall and the movable stopper tip 12.
However, the liquid is not milled by the O-ring 14' and the particulate matter is not milled by the scraper 15', and then packed into the column.

In this case, since the scraper 15° is disposed below the 0-ring 14°, it is possible to completely prevent particulate matter from entering between the 0-ring and the column wall.

On the other hand, in order to ensure the sealing performance of the O-ring 14 even when the movable stopper is raised, the scraper 15 is placed above.
It is now possible to maintain perfect sealing performance.

3.5 Example according to the present invention Next, using the column packing apparatus according to the present invention, a resin gel whose main performance is ion exchange (gel for Ronpak KS-802) was packed into a liquid chromatography column, and an oligosaccharide was sampled. An example will be described in which various types of sugars are separated.

3.5.1 Number of Theoretical Plate When the gel was packed into each of the columns of 20 mmφ and 50 mmφ, the number of theoretical plates was measured while changing the packing load in the range of 20 to 100 ml/min. The results are shown in FIG. 3.

In Figure 3, a 5haφ column has a packing load of 25mm/min and a packing load of 13,000 plates/m, and a 20mmφ column has a packing load of 90mm/win and a packing load of 22,000 plates/m.
The number of theoretical plates is .

Incidentally, in the conventional method using the gas pressurization method, there is a limit to the filling pressure, which is about 4,000 plates/plate.

3.5.2 Oligosaccharide separation/preparation performance Figure 4 shows the preparative separation results when gel was packed into an 80 mmφ column using the conventional method.

In this case, since the upper limit of the working pressure is 20 kg/cm'G, the maximum flow rate of the solution solution is 100 m1/11 in, and the number of theoretical plates is 7,000 plates/ffl, and it takes about 10 minutes to complete the fractionation.

FIG. 5 is a chromatograph obtained by filling an 80 mmφ column with the same gel according to the method of the present invention and performing fractionation under the same conditions as the conventional method. Eluent flow rate 1001/
When the pressure is 30 kg/cm'G, the number of theoretical plates is 20,000 plates/I11, and it can be said that the separation performance is improved compared to the conventional method.

FIG. 6 is a chromatograph obtained when the same column as in FIG. 5 was pressurized to a pressure of 90 kg/cm'G, which cannot be used in the conventional method, and the eluent flow rate was set to 300+++ I/min.

In this case, it takes about 3 minutes to complete the separation, and the separation performance is the same as in the case of FIG. 5, that is, a considerable improvement over the conventional method.

3.6 Effects of the present invention The method and apparatus of the present invention make it extremely easy to pack a column for liquid chromatography, and at the same time increase the number of theoretical plates in a column packed in this way.

[Brief explanation of drawings]

Figure 1 is a schematic configuration diagram of the packing device according to the present invention, Figure 2 is a configuration diagram of the seal section and scraper, Figure 3 is a diagram showing the relationship between column packing load and the number of theoretical plates, and Figure 4 is a diagram of the relationship between column packing load and the number of theoretical plates. A diagram showing the results of fractionation using a column packed according to the conventional method, and FIGS. 5 and 6 are diagrams showing the results of fractionation using the column packed according to the present invention. 1-column, 2-movable stopper, 3-movable stopper moving base, 4A
, 4B-rotating shaft, 5-electric motor, 8-column pedestal, 7-
Reduction mechanism, 11-movable stopper shaft, 12-movable stopper tip, 13
-Spacer, 14. +4°-0-ring, 15.15
-Scraper, 16-Filter, 1? -771 moves,
18-Manufacturing patent applicant Showa Denko Co., Ltd. Representative Patent attorney Sei Kikuchi- Figure 3 Figure 4 ELuent: H2C FIOwrate + 100 ”'/=nPress
ure: 20 kg/cn2Q Sarnpte:
OLigosCLcchcLrtde50 2
．． 5.5 7.5 10 (mm) (min) (ffLLrL) P system amendment (method) 1985 lρ month f day

Claims (1)

[Claims] 1. Packing of a column for liquid chromatography, characterized in that a movable stopper attached to one inlet of the column is pressurized into the inside of the column by a mechanical conduction mechanism to pressurize the packing material. Method. 2. A liquid chromatography column packing device characterized by having a movable stopper, a mechanical conduction mechanism for press-fitting the movable stopper into the column, and a pedestal for fixing the column. 4. The method for packing a column for liquid chromatography according to claim 1, wherein the press-fitting load is constant. 5. The packing device for a column for liquid chromatography according to claim 2, characterized in that the movable stopper has a scraper.