JPS5879155A - Gradient device for liquid chromatograph - Google Patents

Abstract

PURPOSE:To perform liquid transportation and mixing exactly in a specified mixing ratio by installing a pump the liquid discharge of which is exact in each flow channel of solvents more than two kinds. CONSTITUTION:With the start of a programmer 5, the pistons of low-pressure pumps 3 and 4 operate in the direction of sucking, and each solvent of solvent tanks 1 and 2 is sucked until it fills the capacity of the cylinder. Next, with the movements of the pistons to extruding, supply of the solvent to the mixer is started. After starting exact transportation of each solvent, a high-pressure pump 8 is driven to transport liquid. Then, when the conditions permit measurement, analysis specimen is introduced by a specimen injection device 11, and the program under the gradient condition that is set up in the programmer 5 is started, and an introduced specimen is separated into components of a plural number by means of a separation column 12. The separated components are introduced into a recorder 14 and data processing device 15 via a detector 13, and there the chromatogram and component concentration are displayed.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is particularly applicable to the fields of chemistry, medicine, biology, etc.
By changing the mixing ratio of different solvents over time, the components in the sample introduced from the syringe were pumped to the separation column using seven high-pressure pumps. The present invention provides a gradient device for liquid chromatography that dispenses .

Recently, high-performance liquid (4C) chromatography has been applied to a wide variety of fields, and fields where gas chromatography has traditionally been used are gradually transitioning to this method.

Depending on the type of sample, separation and analysis may take a long time. Such a sample is a.

In the isocratic elution method, that is, the elution method with an eluent having a uniform composition, the elution of substances that are strongly retained in the separation column is delayed, the peak broadens, the detection sensitivity is reduced, and analysis requires a long time.

Particularly in the case of biological samples, the amount of sample is small, and the molecular weight of the component to be analyzed is comparatively large, so elution from the separation column is delayed, so detection sensitivity must be increased. One possible solution to this problem is to increase the sensitivity of the detector, but it is technically difficult to further improve the performance of currently commercially available detectors.

If a more sensitive detector is applied, However, it is not possible to reduce the time required for wing separation.

For this reason, in recent years, the gradient elution method, in which the composition of the eluent is changed over time to hasten the bathing time of sample components, has come into widespread use, and many manufacturers are already offering elution methods that have a gradient function. Liquid chromatographs are on sale.

There are many of these gradient installations fd: /)
, two or more high-pressure pumps are used to pump each solvent constituting the eluent one by one and collect it in one place,
Method of sending the liquid to the separation column after mixing and stirring, 2) 7
From the moment the high-pressure pump enters the suction operation to the discharge operation, two or more solvent suction lines are connected.
- The on-off valve installed in the column is controlled by a programmer (*Ia L), and a predetermined separation and condensation liquid is prepared (2) and then sent to the separation column.

In the former case, multiple high-pressure pumps provide peace of mind, and the equipment is expensive. Predetermined 11; is not inhaled, and the composition of the solution M fluctuates. It is doubtful that a similar compositional change would occur due to V.

This is a major factor in the analysis.

The present invention has the following features: /F Small/Small Advantages/Points, One Corner, One Eraser, One Solution with a Predetermined Mixing Ratio for Hunting, L, /11.
We provide 7 systems and a means for sending liquid to 1 rice cake and 2 pieces of rice cake.

That is, the present invention uses a programmer to control the liquid feeding -111 of a low-pressure pump installed in the suction line of each *vll liquid solvent to accurately prepare a predetermined +f511f ratio (+f511f) fusing liquid, which is then transferred to seven high-pressure The pump performs suction and discharge to send liquid to the sample injector, column 11, and detector.
(j, damono desu.

3- FIG. 1 is an explanatory diagram showing an example of the configuration of the apparatus of the present invention, and the configuration and operation thereof will be explained below.

/1.2 is the solvent tank, 3 is the low pressure pump, j is the programmer, O is the mixer, 7 is the deaeration tank, is the high pressure pump, ri is the pressure filter, /θ is the drain valve, // is the sample injection /2ff separation column, /3 detector, /4t is recorder, /j day solvent tank/, 2 (A: (Each solvent that makes up the ri bath separation liquid is input. , Rikeso h
The flow rate range is about θ, θ/~2θ7/mj, n,
A reciprocating piston pump having a discharge pressure range of about θ to 2/:, 9/ca is preferable, and other pumps such as a squeezing pump, a diaphragm pump, a centrifugal pump, and a gear pump are also used.

The rotational speed of the motor of these pumps is controlled according to the eluent flow setting, but the flow rates of the low-pressure pumps 3 and 3, which pump the solvent, are controlled by the eluent ratio set in the programmer 3. The sum of the flow rates is usually constant.

"Programmer" is for controlling the low pressure pump 3, and it sets the flow of solution H1 solution 111, the solution H1 addition fIi at the beginning and end of the gran end, error 111, the time required for the gradient, and the gradient method. It has gradient start, gradient hold and reset functions that keep the concentration constant for a certain period of time during the gradient, and an eluent concentration display function.

Mixer otsu iqJ: , , 2 flli j so, mouth α
) It is used to prepare the eluent by mixing solvents.
2 +'lJ and an example is shown in FIG. Fig. 2 r Partial introduction of an example of an example of a mixer used one by one t1 [Top view;
t, le. The body of the mixer 6 is l'N: cylindrical with a volume of θ0.2 to Δ1lI11!, excluding the volume of the stirrer/ton at medium heat. a mixing chamber /9, a plurality of solvent inlets / and /t' communicating with m pieces of the chamber /9, # syneresis outlet 2θ leading to the other side;
and a plug 22 for closing off the mixing chamber, which may be made of synthetic resin, metal, glass, ceramic, or the like.

The main body/B is installed so that the central axis of the cylindrical mixing chamber/9 is horizontal.

Stirring 1,000/2 is a magnetic (+ coated with synthetic resin, glass, ceramic, etc.) - shaped material with center 41 (1
It is preferable that one end of the direction has a fin shape (which increases the stirring effect of the medium), and the fin part is arranged on the solvent introduction 11 side of the migizer.
Rotated by f.

Degassing N*7 is for separating bubbles generated by mixing the solvents from the eluent.In the device of the present invention, the solubility angle of the solvent vapor is changed to It may change and bubbles may occur. An example is shown in FIG.

The eluent mixed and stirred by the mixer B (d gas 7U and the TRC 1 resin tube 23 (r) is passed through the degassing tank 2
Enter.

At this time, it is more effective during gas-liquid separation if the tip of the resin tube 23 is brought into contact with the inner wall of the degassing tank 2 and the solvent is gently introduced into the degassing tank.

The eluent flows along the inner wall of the degassing tank 2 and exceeds the capacity of the liquid chamber 2. Ili:
It is discharged as waste liquid from the pipe inlet. ”] Koki/fg frl
Easy to exhaust, exhausted from 2f.

There is only one in the liquid reservoir 2B. In order to prevent the degree of a of the mixed solution from changing due to pinching, it is desirable that the volume is small, <, usually θ, 3 to .2
Me is preferable Ll2), Kikaga is crow, synthetic resin,
Metal etc.

A high-pressure pump is usually a high-speed liquid pump, and is used as a built-in reciprocating screw. is 3θ~jθθ/(9/ c, H is t+, id:'JL<, 1ζ1! Formula is d single plunger pop, double plunger pump or ia: l ripple grunge i - It can be done without using any pumps etc.

Pressure filter 21 is used to reduce pulsation during liquid transfer with the high-pressure pump, and has an internal volume of θ, 3~jml.
A flat tube type, a plutono tube type, a bellows type, a liquid compression type, etc. are used, and the type of pressure filter 90'i detector and the concentration V of the target substance a to be analyzed (therefore, it may not be necessary to use it in some cases).

Drain valve/θ is a three-way valve that facilitates the extraction of the eluent flowing through the system.When changing the type of solvent, the pressure in the high pressure pump and subsequent channels becomes abnormal (e.g., when the pressure becomes high). Open the valve and pour out the liquid.

For the sample injector//after, various known ones are used.

For example, sample injectors include a rotary valve type, a bypass flow path type, and the like. t as shunt column/3
7: j distribution force, l! , adsorption type, ion exchange type, etc.

The detector/3 includes an ultraviolet detector, a refractive index detector, a fluorescence detector, an electrochemical detector, etc., and is used depending on the form of the substance to be measured in the sample.

8- Next, using the device of the present invention configured in this way,
Cases in which liquid chromatography analysis is performed will be explained.

Two types of solvents A,] 33, 2, 2, 3, 5, 7, 7, 7, 7, 7, 7, 7, Volume medium A1 is a fast elution solvent, and the gas density ratio is 1 (
′υ) The small solvent is 13, and the solvent tank /(/′−M 4
To! :' A, clear medium, 2 solvent B chopsticks - change.

Low pressure pops 3 and 0, 7 and 1, 3, 11, 3, 3, 3, 3, 3, 3, 1, 2, 1, 2, 1, 2, 1, 1, 2, 3, 3, 1, 2, 3, 3, 3, 3, 1, 2, 1, 2, 1, 2, 1, 2, 1, 2, 1, 2, 1, 2, 3, 1, 2, 1, 2, 3, 1, 2, 3, 1, 2, 1, 2, 3, 1, 2, 1, 2, 1, 2, 3, 3, 3, 3, 3, 3, 3, 3, 3, 1, 1, 2, and 1 in them, respectively, and 1 to 1 to 1 to 1.
゛C yo 1 noi: (J'lE;iζ''"3 JL・"
,0・gσ) Pikutton moves in the direction of pull-, the solvent,
hllH/ , :l (N) Each dG/A1 is cylinder capacity -!it k 1If77 minus 1 and J'+
Ru.

After that, the pistons of the low-pressure pops 3 and 3 move in the pushing direction 5, and the supply of solvent to the millimeter B is started, but the liquid composition in the miller 6 does not change from the predetermined concentration.
V (, while one pump is sucking the solvent, the other pump is
1) Stop the pump as well. Koh is single-edged (7
] If one pump is suctioning solvent and the other pump is pumping normally, the solvent concentration inside the mixer will temporarily drop below the set value, making it impossible to perform proper gradient elution. be.

Ratio to the set flow rate of the high-pressure pump that sends liquid to /θ~20
It is desirable to increase the amount by %, and the excess eluent should be degassed + @
It is discharged as waste liquid from one end of 2.

After the low-pressure pump 3 and the pump begin to accurately feed the solvent, the high-pressure pump and the high-pressure pump are driven to carry out liquid feeding.

After the entire system is ready for measurement, the sample injector//Vc
Therefore, introduce the analysis sample and start the program with the gradient conditions set in the programmer at the appropriate time.

The concentration of solvent B gradually increases from the initial concentration, e.g. /θ volume %, until it reaches a final concentration, e.g. Return to initial concentration.

The sample introduced from the sample injector // is separated into individual or multiple components in the separation column /2, exits the separation column /2 and enters the detector /3. Detector/
3.i) The detection signal of the radar measurement component is recorded by the recorder/g:l
', .J: is introduced into the data processing device 7, and displayed as a chromatogram and component concentration.

As detailed above, according to the present invention, by installing a pump with an accurate flow rate in the flow path of J1 for the two solvents constituting the eluent, When changing the mixing ratio of the eluent over time and continuously feeding the eluent, it is possible to accurately feed and mix the eluent at the predetermined mixing ratio, regardless of the viscosity of each solvent. It is extremely useful as a gradient device for liquid chromatography.

[Brief explanation of drawings]

FIG. 1 is an explanatory diagram showing an example of the configuration of the apparatus fi of the present invention. FIG. 1 is a vertical cross-sectional view of an example of a deaeration tank. /, 2: Solvent tank, 3, Guni low pressure pump,! : Programmer, 3: Mixer, 2: Deaeration tank, Ryoji High Pressure Pump Applicant Mitsubishi Chemical Corporation Representative Patent Attorney Hase - and 7 other people 12-- Sen) Mi 2/d Isamu 3 Figure 5

Claims (1)

[Claims] A storage tank for separately storing one or more different solvents for each of the solvents, two or more low-pressure pumps for sucking and delivering the respective solvents, a device for controlling the discharge amount of each of the low-pressure pumps, and discharge from the low-pressure pumps. a mixer with a stirrer for mixing the respective solvents; a deaeration tank for storing the mixed liquid and removing air bubbles generated by the mixing; and a deaeration tank for sucking the mixed liquid stored in the deaeration tank. A gradient device for liquid chromatographs that consists of a high-pressure pump that sends liquid to the column of a liquid chromatograph.