KR101740596B1 - System for filling column of high performance liquid chromatography - Google Patents

Abstract

The present invention relates to a high performance liquid chromatography column filling system. More specifically, provided is a high performance liquid chromatography column filling system which comprises: a tank part storing a solvent; a pump including an inlet and an outlet and filling the column with the solvent through the outlet by driving a cam; and a controlling part taking control to linearly raising filling pressure for the solvent by the pump in proportion to a time.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a high performance liquid chromatography column charging system,

The present invention relates to a filling system for high performance liquid chromatography columns, and more particularly to a filling system for high performance liquid chromatography columns capable of uniformly filling the filler throughout the column.

High Performance Liquid Chromatography (HPLC) is a type of liquid chromatographic reagent in which the moving phase of a sample is liquid, which is widely used because of its high efficiency and resolution. Liquid chromatography is a technique for injecting a sample into an HPLC column and flowing a liquid mobile phase to separate each component of the sample using the difference in affinity between the stationary phase and the mobile phase.

The HPLC column is prepared by high pressure injection of a chromatographic medium (or filler), which is a stationary phase in slurry state, using a pump in a metal tube (or an empty column).

Specifically, in the step of filling the filler into the common column, a filler is injected into the pressurized solvent, and then a substitution solvent is further injected to replace the pressurized solvent with the mobile phase solution.

Such a column filling process is, for example, filling a 3 占 퐉 or 5 占 퐉 filler uniformly at a high density in a SUS tube having an inner diameter of 2 to 4.6 mm. When the filling density of the column is low, the sample spreads in the pores in the column, Even if a column having a lower number of columns is formed and a column is packed at a high density and a sufficient theoretical number of columns is obtained, the shape of the eluted peak will be poor unless the packed bed in the column is uniform. In other words, the leading part of the peak is pulled up, and the rear part is attracted to tailing.

On the other hand, the pump used in the column filling process is an air drive constant-pressure pump or an electric drive constant-pressure pump. Although the air drive constant-pressure pump can feed high pressure, since the pressure is not constantly transmitted due to the large pulsation, , An electric drive constant pressure pump capable of delivering a constant pressure is often used.

However, when the pressure of the conventional static pressure pump is raised, the charging pressure is increased as shown in the graph of FIG. That is, although the constant pressure is increased step by step, if the method is followed, the defect rate of the column can not be predicted. That is, there is a difference in the defect rate depending on the number of step-up steps, the time required to raise the pressure step by step, and the skill level of the work.

SUMMARY OF THE INVENTION Accordingly, it is an object of the present invention to provide a high-performance liquid chromatography column filling system capable of uniformly filling a column filler, .

According to an aspect of the present invention, there is provided a system for filling a high performance liquid chromatography column, the system comprising a tank portion containing a solvent, an inlet port and a discharge port, the chamber being filled with the solvent through the discharge port by driving a cam And a controller for controlling the charge pressure of the solvent through the pump to rise linearly with time. The present invention also provides a system for charging a high performance liquid chromatography column.

According to one embodiment, the tank portion is disposed in a first tank containing a pressurized solvent, a second tank containing a substitution solvent, and a pipe between the first and second tanks and the inlet, And a switching valve for switching the flow path formed between any one of the first and second tanks and the suction port.

According to one embodiment, the control unit may transmit a flow path switching control signal to the switching valve when a predetermined valve switching time is reached.

According to one embodiment, the apparatus further comprises a pressure gauge for measuring the pressure of the pipe between the discharge port and the column, wherein the control unit is configured to feed back the measured pressure in real time from the pressure gauge, The pump can be controlled to rise linearly.

According to one embodiment, the pair of plungers disposed on both sides of the rotation axis of the cam reciprocally move reciprocally, so that the pump can have a suction port and a discharge port on each side.

According to one embodiment, the pressure gauge may be installed in only one of the pipes connected to each of the two outlets.

Further, the present invention is a method for filling a high performance liquid chromatography column, wherein the pump having the inlet and the outlet includes the step of filling the column with the solvent contained in the tank through the outlet by driving the cam, The filling step may control the control part to linearly increase the filling pressure of the solvent through the pump with time.

According to one embodiment, the pump comprises a step of filling the column with a pressurized solvent contained in a first tank during a pressurized solvent charging time, and the pump further comprises the step of supplying a displacement solvent, contained in the second tank, Wherein a switching valve is formed between an inlet of the pump and an outlet of the first tank between a step of filling the pressurized solvent and a step of filling the displacement solvent, in accordance with a control signal of the control unit And changing the flow path from the first flow path to the second flow path formed between the inlet of the pump and the outlet of the second tank.

According to the present invention, the filler is uniformly charged by linearly filling the filler into the HPLC column over time, thereby improving the performance of the high performance liquid chromatography column, ensuring the reproducibility of the HPLC column, Can be lowered.

In addition, it is possible to charge the two columns in parallel by driving one cam, so that a column having the same performance can be manufactured.

1 is a graph showing a charging pressure rise of a conventional static pressure pump.
2 is a configuration diagram of a high performance liquid chromatography column charging system according to an embodiment of the present invention.
3 is a graph showing a charge pressure rise of a static pressure pump according to an embodiment of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

HLPC Column  Charging system

2 is a configuration diagram of a charging system for an HLPC column according to an embodiment of the present invention.

2, the HLPC column filling system 100 according to an embodiment of the present invention includes tank portions 121 and 122, a pump 110, a controller 170, switching valves 131 and 132, And a pressure gauge (160).

However, it is needless to say that the components shown in FIG. 2 are not essential, so that a charging system of an HLPC column having more or fewer components can be implemented.

Hereinafter, each component will be described.

The tank portions 121 and 122 may contain substances to be introduced into the columns 151 and 152 by means of the pump 110 as means for accommodating various substances therein. That is, it is possible to store the liquid substance to be filled in the columns 151 and 152.

According to one embodiment, the tank portions 121, 122 may include a plurality of tanks to contain different materials. 3, the tank portions 121 and 122 may include first tanks 121a and 122a containing a pressurized solvent and second tanks 121b and 122b containing a replacement solvent.

In the present invention, a pressurizing solvent or a substitution solvent is not particularly limited, but the pressurizing solvent may be methanol, dichloromethane, acetone or the like, and a substitution solvent may be a mixture of dichloromethane and methyl alcohol.

When the filling system 100 of the HLPC column according to an embodiment of the present invention includes a plurality of tanks, a branch portion on the pipes P11 and P12 connecting between the outlets of the respective tanks and the suction port of the pump 110 And may include switching valves 131 and 132.

The switching valves 131 and 132 are disposed in the pipes P11 and P12 between the first tanks 121a and 122a and the second tanks 121b and 122b and the suction port of the pump 110, The flow path can be changed so that the target substances introduced into the columns 151 and 152 are switched to each other according to the signal. Specifically, the first flow path formed by the suction port of the pump 110 and the first tanks 121a and 122a and the second flow path formed by the suction port and the second tanks 121b and 122b are switched .

The pump 110 is a device for transporting fluid through a pipe using a pressure action. The pump 110 may include a suction port for sucking the fluid to be transported and a discharge port for discharging the sucked fluid. Accordingly, the suction port is connected to the outlets of the tank portions 121 and 122 through the pipe, and the fluid contained in the tank portions 121 and 122 can be drawn into the suction port by the suction force generated by the pump 110. [ The discharge port is also connected to the inlet of the column 151, 152 (or the packer 141, 142) through the pipe and the sucked fluid pressurized by the toe output generated by the pump 110 flows into the column 151 , 152 (or packers 141, 142).

At this time, the pump 110 can perform the pumping action in various ways, but it is preferable to use a plunger, a piston or the like (in this specification, the plunger, Is called a reciprocating motion in the cylinder. That is, according to one embodiment, as shown in Fig. 2, three cam plates which are eccentric in different directions on one rotation axis can be combined, and three plungers Are driven to have a phase difference of 120 DEG with each other, so that pulsation is minimized.

According to an embodiment of the present invention, the plunger reciprocating by the driving of the cam may be disposed on both sides of one rotation axis. That is, as shown in Fig. 2, a pair of two plungers disposed on both sides (a side and b side) by rotation of one cam plate can reciprocate simultaneously, so that a total of six plungers are one It is possible to reciprocate by the rotation of the rotating shaft of the motor. With the symmetrical cam structure, two pump heads 112 and 113 that perform a pump action by reciprocating in a phase difference of 120 degrees can be provided on both sides of the pump 110, and the pump heads 112 and 113, 113 have an inlet and a discharge port so that the same solvent contained in one tank can be charged in parallel to two columns 151, 152 (or packagers 141, 142 at the same time) Columns 151 and 152 may be fabricated.

Meanwhile, the control unit 170 may perform various calculations or process various information as means for controlling the operations of the components included in the HLPC column charging system 100, particularly, the pump 110. [ At this time, the control unit 170 may include a control program for controlling or driving at least one configuration included in the charging system 100 of the HLPC column, and a storage unit (or a storage medium) have.

The control unit 170 controls the operation of the pump 110 so that the solvent contained in the tank units 121 and 122 is supplied to the columns 151 and 152 by the pump action of the pump 110. [ , It is possible to control the charging pressure to rise linearly with time.

Conventionally, as shown in Fig. 1, the charging pressure is controlled so as to be input from the user or to rise stepwise to a predetermined target pressure. Specifically, after maintaining the first filling pressure for a predetermined time (until reaching a predetermined flow rate specifically), the pressure is increased from the first filling pressure to the second filling pressure, and the second filling pressure is maintained for a predetermined time The process repeats. As described above, conventionally, there has been a problem that the pressure increasing timing is not uniform because the pressure increasing reference is increased step by step as the flow rate (ml / min) is increased.

However, the controller 170 according to the present invention controls the pump 110 to linearly increase the filling pressure of the solvent with time. That is, as shown in FIG. 3, the controller 170 may generate a control signal such that the charge pressure linearly increases up to the target pressure over time.

At this time, the control unit 170 may generate the control signal based on the signal output from the pressure gauge 160 for measuring the charging pressure to control the charging pressure of the pump 110 to rise linearly. That is, the control unit 170 feedbacks the measured charging pressure from the pressure gauge 160 in real time and controls the charging pressure of the pump 110 based on the feedback, It is preferable to control so as to follow the straight line. Here, the rising slope of the charging pressure can be input from the user or preset.

Since the fillers are charged in the columns 151 and 152 by the linear booster method, the filler is constantly charged from the bottom to the upper layer, so that the filling is uniform.

Meanwhile, according to one embodiment of the present invention, as described above, the pressure gauge 160 may be included.

The pressure gauge 160 measures the pressure of the fluid flowing along the pipe. In order to control the filling pressure based on the pressure measured by the pressure gauge 160, And transmits the generated signal to the control unit 170.

The pressure gauge 160 may be of various kinds depending on the method of measuring the pressure and the like, but the kind thereof is not particularly limited.

The pressure gauge 160 is disposed in the piping P12 and P22 between the columns 151 and 152 (or the packers 141 and 142) and the pump 110 (more specifically, the discharge port) It is preferable to measure the pressure of the solvent.

If the pump 110 has the pump heads 112 and 113 having the suction port and the discharge port in parallel on both sides thereof, the pressure gauge 160 may be connected to the piping (not shown) connected to the respective discharge ports P12, and P22. However, since the solvent discharged from each of the discharge ports by the two plungers driven by one cam interlocked with each other has the same flow rate or pressure, it is installed in only one of the pipes .

Hereinafter, the operation of the HLPC column charging system according to an embodiment of the present invention will be described in detail with reference to FIG.

The charging system 100 of the HLPC column according to an embodiment of the present invention may include at least one of a target pressure, a pressurized solvent use time, a valve change time, and a substitute solvent use time through an input device (not shown) One or a combination thereof.

Accordingly, the control unit 170 controls the pump 110 to raise the filling pressure up to the target pressure, and to linearly raise the input pressure according to a predetermined input charge pressure rising slope. In this process, the control unit 170 controls the filling pressure of the pump 110 so that the filling pressure linearly increases along the slope based on the filling pressure measured in real time through the pressure gauge 160.

When the pump 110 has reached the target pressure, the target pressure is kept constant until the time of using the pressurized solvent or the valve switching time. When the valve 110 reaches the valve switching time, the control unit 170 causes the switching valves 131, Signal. The suction ports of the pump 110 are connected to the outlets of the second tanks 121b and 122b by the switching valves 131 and 132. [

Accordingly, the pump 110 operating at the target pressure discharges the replacement solvent contained in the second tanks 121b and 122b into the columns 151 and 152 during the replacement solvent use time.

Example

The columns 151 and 152 have a size of ID4.6 mm x Length 250 mm and the pressurized solvent contained in the first tanks 121a and 122a is C18, 5 탆 and 120 Å, and the displacements contained in the second tanks 121b and 122b The solvent is 60% AcCN.

The evaluation data according to the detection result using UV 254 nm when 20 나 of naphthalene was injected at a flow rate of 1 ml / min at 35 캜 is shown in Table 1 below.

At this time, the pressurized solvent and the substitution solvent were charged into the column according to the graph of FIG. Specifically, the pressurized solvent was linearly increased from 0 to 8000 psi for the first 136 seconds, then maintained at a charge pressure of 8000 psi constantly for 1382 seconds, and the changeover valve was operated at 1382 sec to change the displacement solvent from 832 psi The columns 151 and 152 were filled.

Here, N and As of the naphthalene peak in the 60% AcCN mobile phase were evaluated. The criterion for the determination was N = 25,000 or more and the peak symmetry As was 0.95 to 1.1.

As shown in Table 1, the acceptance rate is 90%.

Comparative Example

When the pressurized solvent and the substitution solvent were charged according to the graph of FIG. 1 under the same conditions as in the above example, the evaluation data are shown in Table 2 below.

Specifically, the pressure of the pressurized solvent was maintained at 220 psi until the flow rate reached 4 ml / min. The flow rate was 4 to 8 ml / min, 8 to 12 ml / min, 12 to 16 ml / The filling pressure was maintained at 2200 psi, 3300 psi, 4520 psi, 5680 psi, 6840 psi, 8000 psi in 16 to 19 ml / min and 19 to 181 ml / min, respectively. When the flow rate was 181 ml / min, The charge pressure was maintained at 8000 psi up to 381 ml / min.

As shown in Table 2, the acceptance rate is 70%, and the acceptance rate is 20% lower than the above embodiment.

That is, according to one embodiment of the present invention, filling the filler linearly with time in the HPLC column has the effect of uniformly filling the filler, thereby lowering the defect rate.

The preferred embodiments of the present invention have been described in detail with reference to the drawings. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the appended claims.

Accordingly, the scope of the present invention is defined by the appended claims rather than the detailed description, and all changes or modifications derived from the meaning, range, and equivalence of the claims are included in the scope of the present invention Should be interpreted.

100: Charging system of HLPC column 110: Pump
111: Cam 121a, 122a: First tank
121b, 122b: second tank 131, 132: switching valve
141, 142: Packer 151, 152: column
160: pressure gauge 170:

Claims (8)

In a high performance liquid chromatography column filling system,
A tank portion containing a solvent;
A pump having a suction port and a discharge port, the pump driving the cam to fill the column with the solvent through the discharge port; And
A controller for controlling the charging pressure of the solvent through the pump to rise linearly with time;
≪ RTI ID = 0.0 > 1, < / RTI > The method according to claim 1,
The tank portion
A first tank containing a pressurized solvent;
A second tank containing a displacement solvent; And
A switching valve disposed in a pipe between the first and second tanks and the suction port for switching the flow path formed between any one of the first and second tanks and the suction port according to a control signal of the control unit;
≪ RTI ID = 0.0 > 1, < / RTI > 3. The method of claim 2,
Wherein,
And when a predetermined valve switching time is reached, a flow path switching control signal is transmitted to the switching valve. The method according to claim 1,
A pressure gauge for measuring a pressure of the pipe between the discharge port and the column;
Further comprising:
Wherein the control unit feeds back the pressure measured in real time from the pressure gauge and controls the pump so that the filling pressure of the solvent linearly increases with time. 5. The method of claim 4,
Wherein the cam has a pair of plungers disposed on both sides of the rotation axis and reciprocatingly reciprocally moved so that the pump has a suction port and a discharge port on both sides thereof. 6. The method of claim 5,
The pressure gauge includes:
Characterized in that it is installed in only one of the piping connected to each of the two discharge ports. In a method for filling a high performance liquid chromatography column,
The pump having the suction port and the discharge port includes a step of filling the column with the solvent contained in the tank portion through the discharge port by driving the cam;
, ≪ / RTI &
Wherein the filling step controls the controller to linearly increase the filling pressure of the solvent through the pump with time. 8. The method of claim 7,
The pump comprising: charging the column with a pressurized solvent contained in a first tank during a pressurized solvent charging time; And
The pump comprising: charging the column with a displacement solvent in a second tank during a displacement solvent charge time;
, ≪ / RTI &
Wherein a switching valve is provided between the step of filling the pressurized solvent and the step of filling the displacement solvent in a first flow path formed between an inlet of the pump and an outlet of the first tank in accordance with a control signal of the control unit, And a second flow path formed between an inlet of the first tank and an outlet of the second tank;
≪ / RTI > further comprising the steps of:

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