KR20190043030A - Sample Introduction System of Liquid Chromatography - MALDI Mass Spectroscopy Manufactured by 3D Print - Google Patents

Abstract

The present invention relates to a sample introduction system of a liquid chromatography-MALDI mass spectrometer (LC-MALDI MS). More particularly, the present invention relates to a sample introduction system of an LC-MALDI MS including a component with a polymeric material manufactured by using a 3D printer, and a manufacturing method thereof.

Description

MALDI mass spectrometry (MALDI) mass spectrometry (MALDI) mass spectrometry (MALDI) mass spectrometry (MALDI)

More particularly, the present invention relates to a liquid chromatography-mercury-mass spectrometer (LC-MALDI-MS) which can be produced more simply and economically by using a 3D printer, And a method of manufacturing the same.

Mass spectrometers are generally designed to measure the mass of a compound and are designed to determine the molecular weight of a compound by ionizing the analyte and measuring the mass-to-charge (m / z) of the analyte . As methods for ionizing a compound, there are known an electron ionization method using an electron beam, a method of colliding atoms at a high speed, a method using a laser, and a method of spraying a sample in an electric field.

As a method for mass analysis of a biochemical substance having a macromolecular mass such as a protein and a nucleic acid, a MALDI-TOF MS (Laser-Assisted Laser Desorption Ionization Time of Flight Mass Spectroscopy ), And a mass spectrometer using the mass spectrometer has recently been developed and widely commercialized. Mali mass spectrometry is very useful for analyzing various materials including proteins, peptides, sugar chains, lipids, and nucleic acids.

The mass spectrometry using Maldis is described in detail. First, a small amount of a matrix solution is applied to a metal plate, and the sample solution to be analyzed is dropped and dried. Later, when the matrix and the crystallized sample are irradiated with a laser, the sample can be desorbed / ionized with the aid of the matrix.

When an electric field is applied between a metal plate on which the sample is placed and a sensor for analyzing the mass, the ionized sample moves to the sensor due to the potential difference of the electric field. When the amount of charge of the sample is known, The mass of the sample can be analyzed with the parameters.

On the other hand, liquid chromatography is a method of separating complex analytes from a liquid phase using the physical properties of the analytes, and is used as a pretreatment process for analyzing analytes such as samples extracted in vivo. Such liquid chromatography (MALDI) mass spectrometer can be obtained by combining the above-described mass spectrometry method with a Mali mass spectrometry method.

In a liquid chromatography (LC) -maldi (MALDI) mass spectrometer using the liquid chromatography as a pretreatment process, a sample and a matrix solution continuously flowing in a liquid chromatography are mixed to analyze an analyte with the Mali mass spectrometer It should be loaded onto a specially coated sample plate, and then a sample plate is placed in a Mali mass spectrometer and irradiated with a laser to ionize it. Due to the discontinuous procedure inherent in this Maldisphere mass spectrometer itself, a special device is required to load the sample and matrix solvent continuously flowing in the liquid chromatography on the sample plate for a volume of several μL.

That is, in order to load the sample plate, a commercially available microsyringe pump is generally used as a part of the apparatus, in order to mix a sample and a matrix and inject a sample / matrix mixture of several μL per minute. The microsyringe pump included in the LC-MALDI mass spectrometer is capable of controlling the speed according to the microsyringe standard, but its price is very high, (LC) -Maldi mass spectrometer from a manufacturer of a specific liquid chromatography (LC) -Maldi mass spectrometer to obtain a proper A / S It has limitations.

On the other hand, as a technique related to such liquid chromatography-MALDI mass spectrometry, Japanese Patent Publication No. 5958288 (2016.07.01) discloses that a sample containing a substance to be analyzed is added to liquid chromatography using a mobile phase containing salt, A technique relating to liquid chromatography-mass spectrometry including obtaining an eluate and adhering the resulting eluate to a matrix-assisted laser desorption ionization (MALDI) mass spectrometry using an organic compound having a phosphonic acid group as a matrix additive without desalting No. 10-2017-0042610 (Apr. 19, 2017) relates to a super-clean autosampler for mass spectrometry including syringe delivery, and more particularly to a super-clean autosampler for fluid communication with a first valve and a first valve A valve assembly including a second valve, wherein the valve assembly is actuated by one or more syringe pumps Through the flow of the working fluid it has been described with respect to the description of the system for delivering samples, chemicals and one or more of the standard.

However, in manufacturing the parts of the sample introduction device of the liquid chromatography (LC) -maldi (MALDI) mass spectrometer, it is possible to make the result not different according to the skill of the person and the performance of the equipment, Liquid chromatography (LC) -molded, which is manufactured using a method of minimizing the adhesive to minimize the manufacturing error while using the parts printed on the 3D printer without using any post-processing and to facilitate self-maintenance, It is necessary to develop a device capable of introducing a mixed solution obtained by mixing a sample and a matrix from liquid chromatography (LC) in a mass spectrometry (MALDI) mass spectrometer.

Japanese Patent Publication No. 5958288 (Jul. 2016) Published Japanese Patent Application No. 10-2017-0042610 (Apr. 19, 2017)

The present invention relates to a method of mixing a sample and a matrix from a liquid chromatography (LC) in a liquid chromatography (LC) -maldi mass spectrometer used to inject a matrix solution using a component made using a 3D printer And to a device capable of introducing the obtained mixed liquid.

The present invention also relates to a method of producing an apparatus capable of introducing a mixed liquid obtained by mixing a sample and a matrix from liquid chromatography (LC) in a liquid chromatography (LC) -maldi (MALDI) mass spectrometer.

In order to achieve the above object, the present invention provides a syringe pump; A syringe mountable on the syringe pump; A first sample route pipe connected to the syringe to introduce a MALDI mattress solution (A); A second sample root tube into which a solution (B) containing a sample separated and eluted in liquid chromatography (LC) is introduced; A mixer for mixing the solution of the first sample route tube with the solution of the second sample route tube; A sample solution for discharging the mixed solution of the MALDI mattress solution (A) and the solution (B) containing a sample eluted and separated in liquid chromatography (LC) from the mixer to a sample plate of a MALDI mass spectrometer was introduced tube; A sample plate operating part for adjusting the position of the sample plate; And a sample plate into which the mixed liquid is introduced, wherein the syringe pump includes a syringe support for supporting a syringe, a lead screw nut support for supporting a lead screw nut pushing a piston end of the syringe, Wherein the upper end of the syringe support and the upper end of the syringe support are made of a polymer material, which is manufactured by a 3D printer, and the lower end of the upper end of the syringe support is recessed to surround the outer surface of the syringe Wherein the upper surface of the central portion of the lower end of the syringe support includes a recess to surround an outer surface of the syringe, and at least a portion of the lower surface of the upper portion of the syringe support and at least a portion of the upper surface of the syringe support include a magnet The upper end of the syringe support and the syringe support (LC) -maldi (MALDI) mass spectrometer according to the present invention is characterized in that the lower end of the mass spectrometer can be desorbed by the magnet.

In one embodiment, the lead strap nut support and the motor support may be made of a 3D print and made of a polymer material.

In an embodiment, the upper end of the syringe support and the lower end of the syringe support base may be removable by mounting a plastic block having a magnet therein, respectively.

In one embodiment, the lower end of the syringe support includes a plurality of polishing bar through holes and a plurality of polishing bar supporting wings on both side surfaces thereof. The polishing bar through hole is a portion through which the polishing bar passes, And the one end of the polishing bar through-hole includes an opening that is open to the outside, and the polishing bar supporting wing portion extends from both sides of the lower end of the syringe support base, and the opening portion included in the polishing bar through- The upper end of the polishing bar support vane and the lower end of the polishing bar support vane may include fastening holes that can be fastened with bolts and nuts, respectively.

In one embodiment, the plastic block at the upper end of the syllable support and at the lower end of the syllable support may be a plastic block in which the magnet powder is dispersed inside the plastic surface or a magnet is attached.

In one embodiment, the motor support includes a plurality of polishing bar through-holes and a plurality of polishing bar supporting wings on both side surfaces, one side of the polishing bar through-hole includes an opening that is open to the outside, The supporting wing portion is divided into upper end portions of the polishing bar supporting vanes separated from the opening portions included in the polishing bar through-holes and lower end portions of the polishing bar supporting vanes, extending from both sides of the lower end portion of the motor support, The lower ends of the support vanes may include fastening holes that can be fastened with bolts and nuts, respectively.

In one embodiment, a capillary tube may be mounted at the end of the sample introduction tube.

In one embodiment, the sample plate manipulating portion may be movable in two axes, X and Y, respectively.

In one embodiment, the motor supported by the motor support can determine the movement of the spotting number according to the repetitive motion using the Quotient & Remainder function.

Also, as an embodiment of the present invention, liquid chromatography (LC) capable of separating and separating a sample to be separated in a mixed sample in a liquid state into a dissolved state; A sample introduction device of the liquid chromatography-Maldø mass spectrometer; And a MALDI mass spectrometer. The MALDI mass spectrometry system comprises:

In addition, in an embodiment of the present invention, there is provided an image forming apparatus comprising: a) a top support part supported by a 3D printer and a bottom support part of a syringe support, wherein a central bottom part of the top end part of the syringe support includes a concave part to surround an outer surface of the syringe, The upper surface of the central portion of the lower end of the syringe supporter is manufactured by printing a cylinder support including a recess to cover the outer surface of the syringe with a polymer material; b) inserting a magnet on at least a lower surface of at least a portion of the upper end of the syringe support and at least a portion of the upper surface of the lower end of the syringe support so that the upper end of the syringe support and the lower end of the syringe support can be removed by the magnet; c) manufacturing a motor support supporting a lead screw nut support and a step motor supporting a lead screw nut pushing the piston end of the syringe; d) Combine the syringe support, the motor support supporting the leadscrew nut support and the step motor Manufacturing a syringe pump; And e) coupling the syringe pump with an LC maldispoter, wherein the LC maldispotter is connected to a syringe to introduce a MALDI mattress solution (A); A second sample root tube into which a solution (B) containing a sample separated and eluted in liquid chromatography (LC) is introduced; A mixer for mixing the solution of the first sample route tube with the solution of the second sample route tube; A sample solution for discharging the mixed solution of the MALDI mattress solution (A) and the solution (B) containing a sample eluted and separated in liquid chromatography (LC) from the mixer to a sample plate of a MALDI mass spectrometer was introduced tube; A sample plate operating part for adjusting the position of the sample plate; And a sample plate into which the mixed liquid is introduced. The liquid chromatography (LC) -maldi mass spectrometer is provided with a 3D printer.

At least one of a cylinder support, a leadscrew nut support and a motor support in parts of a sample introduction device of a liquid chromatography (LC) -maldi (MALDI) mass spectrometer is manufactured using a 3D printer, It is advantageous in terms of user convenience such as easy fixing to parts and easiness of fabrication, and it is also advantageous in that the manufacturing cost is low because the parts made of 3D printer and the parts easy to obtain are used.

In addition, the syringe pump of the present invention has been produced with a specific purpose for simultaneously injecting a matrix solution and a sample in a sample introduction device of a liquid chromatography (LC) -maldi (MALDI) mass spectrometer. However, There is an additional advantage that some designs of the part can be changed and easily applied to the syringes of other shapes and sizes.

In addition, the sample introduction device of the liquid chromatography (LC) -maldi (MALDI) mass spectrometer according to the present invention can control not only the nano-liquid chromatograph but also the analytical microfluidic device by controlling the inner diameter of the root tube, It is also compatible with liquid chromatography.

1 is a schematic diagram of a system for connecting a sample introduction device of a liquid chromatography (LC) and a liquid chromatography (LC) -maldi (MALDI) mass spectrometer.
Fig. 2 is a schematic diagram showing parts and structure of a sample introduction device of a liquid chromatography (LC) -maldi (MALDI) mass spectrometer.
3 is a photograph showing the sample plate and the sample plate operating portion.
4 is a schematic view showing a part and an assembling model of the syringe pump.
5 is a schematic view showing the parts and the structure of a syringe supporter made of a polymer material manufactured by a 3D printer.
6 is a schematic view showing the structure of a motor support made of a polymer material manufactured by a 3D printer.
FIG. 7 is a view showing an interface screen including a start position, a total elution time, and a time per spotting as input values for spotting through movement of a sample plate manipulating part according to the present invention.

Hereinafter, the present invention will be described in detail. But may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

In addition, the present invention is capable of various modifications and various forms, and specific embodiments are illustrated in the drawings and described in detail in the following description. It is to be understood, however, that the invention is not intended to be limited to the particular forms disclosed, but on the contrary, is intended to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

Also, in this application, the terms "comprises" or "having", etc. are intended to specify the presence of stated features, integers, steps, operations, elements, parts or combinations thereof, But do not preclude the presence or addition of one or more other features, integers, steps, operations, components, parts, or combinations thereof.

The present invention relates to a syringe pump; A syringe mountable on the syringe pump; A first sample route pipe connected to the syringe to introduce a MALDI mattress solution (A); A second sample root tube into which a solution (B) containing a sample separated and eluted in liquid chromatography (LC) is introduced; A mixer for mixing the solution of the first sample route tube with the solution of the second sample route tube; A sample solution for discharging the mixed solution of the MALDI mattress solution (A) and the solution (B) containing a sample eluted and separated in liquid chromatography (LC) from the mixer to a sample plate of a MALDI mass spectrometer was introduced tube; A sample plate operating part for adjusting the position of the sample plate; And a sample plate into which the mixed liquid is introduced, wherein the syringe pump includes a syringe support for supporting a syringe, a lead screw nut support for supporting a lead screw nut pushing a piston end of the syringe, Wherein the upper end of the syringe support and the upper end of the syringe support are made of a polymer material, which is manufactured by a 3D printer, and the lower end of the upper end of the syringe support is recessed to surround the outer surface of the syringe Wherein the upper surface of the central portion of the lower end of the syringe support includes a recess to surround an outer surface of the syringe, and at least a portion of the lower surface of the upper portion of the syringe support and at least a portion of the upper surface of the syringe support include a magnet The upper end of the syringe support and the syringe support (LC) -maldi (MALDI) mass spectrometer according to the present invention is characterized in that the lower end of the mass spectrometer can be desorbed by the magnet.

The use of the syringe pump is as follows. In order to ionize an analyte using a MALDI mass spectrometer, a mixed solution of a sample and a specific matrix solution is injected to provide a role of loading on a sample plate.

In the present invention, a sample introduction device of a liquid chromatography (LC) -maldi (MALDI) mass spectrometer was manufactured using a component made using a 3D printer, and a sample introducing device Manufacturing and construction of the syringe pump will be described in detail below.

Some parts of the syringe pump are manufactured using a 3D printer. Particularly, 3D printing is used to support parts (stepper motor, polishing rod, lead screw, lead screw nut, etc.) for manipulating syringes and mechanical movements And fixed it to make a single object. Specifically, parts manufactured using a 3D printer are a syringe support for supporting a syringe, a leadscrew nut support for supporting a lead screw nut pushing a piston end of a syringe, and a motor support for supporting a stepper motor. FIG. 4 shows components and an assembly model of the syringe pump. Syringe supporters 121 and 124, lead screw nut supporter 130, and motor supporter 140, which are components output to the 3D printer, are shown in gray.

The syringe support is made for supporting and fixing the syringe.

The upper end of the syringe support includes an upper end portion 121 and a lower end portion 124. The lower end of the upper end of the syringe support includes a recess to surround the outer surface of the syringe, And a syringe is inserted through the through hole formed by coupling the upper end and the lower end of the syringe support base.

Generally, when a syringe pump is used, it is necessary to construct a structure that can easily attach and detach from the syringe because injecting and discharging analytes are frequently performed in the syringe. In a conventional syringe pump, a screw is used to fix the upper and lower ends of the syringe support, and a spring device is used to control the detachment of the two parts.

However, according to the present invention, a method of fixing a component manufactured using a 3D printer using a screw or a spring device is relatively difficult compared to the case of using a metal-based component, and the material of the 3D printer is relatively weak, Friendly printing method was adopted.

Specifically, the upper and lower ends of the syringe supporter are respectively formed by combining magnets, and the upper and lower ends of the syringe supporter are designed to be removable by the magnets 122 and 125.

The upper end of the syringe supporter and the lower end of the syringe supporter may be attached and detached from the upper end of the syringe supporter and the lower end of the syrin supporter, respectively. The plastic block at the upper end of the syllable support and the lower end of the syllable support may be a plastic block in which the magnet powder is dispersed inside the plastic surface or a magnet is attached.

A handle 123 can be attached to the upper end of the syringe support base to enhance the user's convenience. The handle can be easily used by purchasing an acrylic handle or the like.

The lower end of the syringe support base may include a polishing bar through-hole 126 and a plurality of polishing bar supporting wings 128 and 128 'to facilitate detachment of the polishing bar. The polishing bar through-hole is a portion through which the polishing bar passes and is preferably a cylindrical hole whose diameter is 1 to 3 mm larger than the diameter of the polishing bar.

In order to fasten the polishing rod to the polishing bar through-hole, a lower end of the cylinder support has an opening 127 opened to the outside at one side of the polishing bar through-hole, a coupling groove 129 separated by the opening, And an upper end portion and a lower end portion of the polishing-bar supporting blade portion. It is possible to fix the polishing bar to the polishing bar through-hole by tightening bolts and nuts in the fastening groove of the upper end and lower end of the polishing bar supporting wing portion. The coupling grooves of the upper and lower portions of the grinding bar support vane are designed to be easier than the bolt size.

In addition, the lower end of the syringe support may include a bottom support portion that can be fixed to the bottom, and may be formed into a elliptical hole of a relaxed size in consideration of the possibility of errors that may occur during the printing process and the fixing process of the 3D printer. .

In addition, it is preferable that the lead screw nut support and the motor support are made of 3D printing material and made of a polymer material.

The lead screw nut support 130 serves to fix the polishing rod 150 and the lead screw nut 162 and push the piston of the syringe. The leadscrew nut support should slide along the shaft as the lead screw turns by the motor. For this purpose, a linear ball bushing 151 was used for the leadscrew nut support, and a snap ring 152 was inserted between the thin grooves on both sides of the linear ball bushing in order to fix it. This also has the advantage of being able to attach and detach easily so that parts repairing process can be simplified. Then, to fix the leadscrew nut, a hole was made in the 3D printed part to match the position of the three holes in the leadscrew and fixed with a bolt nut.

The motor support is used to fix a stepper motor connected by a coupling rod 161 and a polishing rod and a lead screw used as a movement shaft. The motor support part of the motor support is made to be able to fix the hole to fit the cylindrical protruding part in front of the motor and the four bolt holes that fit the four bolt holes of the motor are designed and firmly fixed.

The polish bar supporting portion of the motor supporter may include a polish bar through hole 141 and a plurality of polish bar supporting wings 143 and 143 'so as to facilitate removal and attachment of the polish bar, The configuration of the rod support vane portion and the method of fastening the polishing rod are the same as those of the support portion of the lower end of the syringe support described above.

In addition, the motor support may include a bottom support portion 145 that can be fixed to the floor, and may be formed into a elliptical hole of a relaxed size, taking into consideration the possibility of errors that may occur during the 3D printer printing process and the fixing process .

The syringe 400 is mounted on a syringe pump and is designed to discharge the matrix solution by a stepper motor.

The first root tube 210 connects the syringe 400 and the mixer 240 to introduce the MALDI mattress solution A from the syringe and discharge the mixed solution to the mixer.

The second root tube 220 serves to introduce a solution B containing a sample separated and eluted in liquid chromatography (LC) and discharge it to the mixer 240.

The MALDI mattress solution discharged from the first sample route pipe may be injected into a mixer by a syringe pump together with a solution containing samples eluted in a liquid chromatograph (LC) connected with a mixer .

The mixed solution of the solution containing the sample and the matrix solution is loaded on the sample plate 310 by the sample introduction pipe 230 connected in the mixer. A capillary may be mounted at the end of the sample introduction tube.

If the mixer includes a first root tube and a second root tube into which the first solution and the second solution can be respectively injected and a third root tube through which a mixed solution of the first solution and the second solution is discharged, The shape is not limited. Specifically, it may be a T shape, a Y shape, or the like. In the present invention, it is preferable to use a T-shaped mixer.

In order to be able to load the sample on the sample plate, one of the sample introduction tube and the sample plate from which the sample comes out should be moved. In order to stably load a small volume of sample, it is preferable that the sample plate is moved rather than moving the sample introduction tube through which the sample is discharged. Thus, the sample plate manipulating portions 310 and 320 are designed in a manner to move the sample plate.

The sample plate manipulating portions 310 and 320 are movable in two axes, X and Y, respectively. As shown in Fig. 3, there is a method in which a lead screw connected to a stepper motor is rotated to move a fixed axis of a lead screw nut.

The movement of the sample plate operating part can be spotted according to the number of spotting cases using a combination of a case structure, a quotient & remainder function, and the like.

As shown in FIG. 7, the values to be inputted for spotting are 'Start Position', 'Total Elution time (min)', 'Time per spot (s)' And a case structure was used to set the starting position of the above input.

For example, according to the initial position and the starting position, there is a case in which movement is required in the row or column direction and a case in which movement is not required. In each case, a case is assigned to the case structure. For example, in order to move from the initial position (1, A) to the start position (1, B), it is not necessary to move in the column direction but move one space in the row direction. That is, if there is a need to move (false) or not (true) for each row and column, then there are four cases. The number of cells to be moved is based on the difference between the initial position and the starting position for each row or column. So, in the above example, the columns are 1-1 = 0, and the rows are 2-1 = 1, so that the column and row are shifted by 0 and 1, respectively.

We used Quotient & Remainder function and case structure to spotting after setting start position. The direction of spotting on the MALDI plate was set to move right, downward, left, and down to minimize movement time during spotting. On the MALDI plate, there are a total of 24 cells in one row. In consideration of the change of direction, two rows, that is, 48 cells, are set as the minimum motions to be repeated, starting from the end column of the start row to the bottom, left, It is a moving motion. The starting line is excluded from repeated motion because the number of spaces to be spotted depends on the starting position.

The quotient of dividing the total number of spotting calculated by the input value by 48 is Quotient (Q), and the rest is Remainder (R). First, the number of spaces to be moved from the starting position to the right is calculated and the spotting for the first row is completed. After that, you decide how many times you want to repeat the repeat motion according to the quotient value. If Q = 0, there are four cases depending on the number of remainders. For example, if the remainder is 1, the spotting ends by moving the lower one, and if the remainder is 25, it ends up spotting one row of MALDI plates and then going down the lower one. In other words, R = 1, R24, R = 25, and 26R47. If the quotiemt is greater than 1, repeat motion as many times as the quotient number, then spotting using remainder in the same way as above. The number of each case is assigned to case structure.

Meanwhile, according to the present invention, liquid chromatography (LC) capable of separating and separating a sample to be separated in a liquid mixed sample into a dissolved state; A sample introduction device of the liquid chromatography-Maldø mass spectrometer; And a MALDI mass spectrometer to construct a liquid chromatography-MALDI mass spectrometry system.

The present invention also provides an image forming apparatus comprising: a) a top support portion supported by a 3D printer and a bottom support portion of a syringe support, wherein a central lower surface of the top portion of the syringe support includes a concave portion to surround an outer surface of the syringe, Fabricating the upper surface of the central portion by printing a cylinder support including a recessed portion to cover the outer surface of the syringe with a polymer material; b) inserting a magnet on at least a lower surface of at least a portion of the upper end of the syringe support and at least a portion of the upper surface of the lower end of the syringe support so that the upper end of the syringe support and the lower end of the syringe support can be removed by the magnet; c) manufacturing a motor support supporting a lead screw nut support and a step motor supporting a lead screw nut pushing the piston end of the syringe; d) Combine the syringe support, the motor support supporting the leadscrew nut support and the step motor Manufacturing a syringe pump; And e) coupling the syringe pump with an LC maldispoter, wherein the LC maldispotter is connected to a syringe to introduce a MALDI mattress solution (A); A second sample root tube into which a solution (B) containing a sample separated and eluted in liquid chromatography (LC) is introduced; A mixer for mixing the solution of the first sample route tube with the solution of the second sample route tube; A sample solution for discharging the mixed solution of the MALDI mattress solution (A) and the solution (B) containing a sample eluted and separated in liquid chromatography (LC) from the mixer to a sample plate of a MALDI mass spectrometer was introduced tube; A sample plate operating part for adjusting the position of the sample plate; And a sample plate into which the mixed liquid is introduced. The liquid chromatography (LC) -maldi mass spectrometer is provided with a 3D printer.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. It will be apparent to those skilled in the art, however, that these examples are provided to further illustrate the present invention, and the scope of the present invention is not limited thereto.

1) Parts manufactured using 3D printer

Some of the structural parts required for the preparation of the sample introduction device of liquid chromatography (LC) -Maldi mass spectrometer were made using a 3D printer (Moai clone, FDM-type, Samdimol, Seoul, Korea). This 3D printer has a heating bed and auto leveling function. Polylactic acid (PLA, Aplus, China) was used as the filament. The 3D printing output was designed with AutoCAD 2017, one of the computer aided design (CAD) software programs, and converted to .STL (Standard Tessellation Language or STEREO-Lithography) files. The .STL file was transformed into G-code using CURA 14.09 and output to a 3D printer.

2) Parts purchased online

In addition to parts manufactured using 3D printers, other parts were purchased online. The structure of the sample introduction device of a liquid chromatography (LC) -Maldi mass spectrometer is shown in FIG. An abrasive rod was used to manufacture the apparatus of the present invention, and the abrasive rod was used for the lower shaft, the upper shaft, and the syringe pump, each having a diameter / length of 8/26, 6/24, 6/14 (mm). The size of the linear ball bush (Aplus, China) used for the lower axis was 8 in inner diameter, 15 in outer diameter, and 45 in length. The lead screw is also used for the lower shaft, upper shaft, and syringe pump, and each diameter / length is 5/20, 5/20, 5/10 (mm). Diameter / length / step size of the stepper motor coupler (Aplus, China) is 15/20/5/5 (mm). A snap ring was used to combine bearings and parts made with 3D printers.

Microsyringes (1705RNR, volume 50 μL, needle size 22s, Hamilton, USA) were used as syringes for the injection of the matrix solution. A needle port (Rheodyne, P / N 9013, USA) was used to connect the micro syringe to the silica capillary.

3) Running software

Arduino and LabVIEW can be used to operate the syringe pump motor. Arduino is used as a digital I / O board to operate the syringe pump, and LabVIEW is used as operating software to control the Arduino Mega 3560 and the stepper motor driver A4988. LINX was used to interface LabVIEW with Arduino's digital I / O board. The software can be used to fine tune the speed and direction of the motor.

4) Composition of sample introduction device of liquid chromatography-Maldodi Mass Spectrometer

Liquid Chromatography - Maldigi There are three axes in the mass spectrometer sample introduction system. The x and y axes to move the sample plate and the Maldi mass analyzer support are the axes acting as syringe pumps. As shown in FIG. 2, the sample introduction device of the liquid chromatography-mass spectrometer of the present invention comprises a syringe pump 100, a solution mixing device 200, a sample plate device 300 and a syringe 400 Respectively. A system for injecting a preprocessed sample in liquid chromatography can be made as in FIG.

Having described specific portions of the present invention in detail, those skilled in the art will appreciate that these specific descriptions are only for the preferred embodiments and that the scope of the present invention is not limited thereto Will be blind. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

100: Syringe pump
121: Syringe support upper part
124: Lower part of syllable support
122, 125: magnets
123: Handle
126: Polishing bar through hole
127: opening
128: upper end of the polishing rod supporting blade
128 'Lower end of the polishing rod supporting blade
129: fastening hole of the polishing rod supporting blade
130: Lead screw nut support
140: Motor support
141: Polishing bar through hole
142: opening
143: upper end of the polishing bar supporting blade
143 'Lower end of the polishing rod supporting blade
145: bottom wing
150: Polishing rod
151: Linear ball bush
152: Snap ring
161: Coupler
162: Lead Screw Nut
200: solution mixing device
210: first sample route pipe
220: second sample route pipe
230: sample introduction tube
240: mixer
300: sample plate device
310: sample plate
320: X-axis mobile unit of the sample plate manipulating unit
330: Y-axis moving unit of the sample plate operating unit
400: Syringe

Claims (11)

In a sample introduction device of a liquid chromatography (LC) -maldi (MALDI) mass spectrometer manufactured using a 3D printer,
A syringe pump;
A syringe mountable on the syringe pump;
A first sample route pipe connected to the syringe to introduce a MALDI mattress solution (A);
A second sample root tube into which a solution (B) containing a sample separated and eluted in liquid chromatography (LC) is introduced;
A mixer for mixing the solution of the first sample route tube with the solution of the second sample route tube;
A sample solution for discharging the mixed solution of the MALDI mattress solution (A) and the solution (B) containing a sample eluted and separated in liquid chromatography (LC) from the mixer to a sample plate of a MALDI mass spectrometer was introduced tube;
A sample plate operating part for adjusting the position of the sample plate; And
And a sample plate into which the mixed solution is introduced,
The syringe pump includes a syringe support for supporting a syringe, a leadscrew nut support for supporting a lead screw nut pushing a piston end of the syringe, and a motor support for supporting the step motor,
Wherein the syringe support base comprises a syringe support upper end of a polymer material manufactured by a 3D printer and a lower end of the syringe support base, the central lower end of the syringe support base includes a concave portion to surround an outer surface of the syringe,
The upper surface of the central portion of the lower end of the syringe support includes a recess to surround an outer surface of the syringe,
The upper end of the syllable support and the lower end of the syllable support are removable by the magnet because at least a lower surface of the upper end of the syllable support and at least a part of the upper surface of the lower end of the syllable support stand each include a magnet, A liquid chromatography (LC) -maldi (MALDI) mass spectrometer. The method according to claim 1,
Wherein the lead-thread nut support and the motor support are made of 3D printing material and made of a polymer material. The method according to claim 1,
Wherein the upper end of the syringe supporter and the lower end of the syringe supporter are each equipped with a plastic block containing a magnet so that the upper end of the syringe supporter and the lower end of the syringe supporter are removable. Introduction device. The method according to claim 1,
The lower end of the syringe supporter includes a plurality of polishing bar through holes and a plurality of polishing bar supporting wings on both side surfaces thereof,
The diameter of the polishing bar through-hole passing through the polishing bar is 1 to 3 mm larger than the diameter of the polishing bar,
One side of the polishing bar through-hole includes an opening that is open to the outside,
The polishing bar supporting vane portion is divided into an upper portion of the polishing bar supporting vane and a lower portion of the polishing bar supporting vane extending from both sides of the lower end portion of the syringe support base and separated by the opening portion included in the polishing bar through-
Wherein the upper end of the polishing-bar supporting vane and the lower end of the polishing-bar supporting vane include fastening holes that can be fastened with bolts and nuts, respectively. The method of claim 3,
Wherein the plastic block at the upper end of the syringe support and at the lower end of the syringe support comprises a sample block of a liquid chromatography-Maldø mass spectrometer, wherein the magnet powder is dispersed in and on the plastic surface, Device. The method according to claim 1,
Wherein the motor support includes a plurality of polishing bar through holes and a plurality of polishing bar supporting wings on both side surfaces thereof,
Wherein one side of the polishing bar through-hole includes an opening that is open to the outside,
The polishing bar supporting vane portion is divided into an upper end portion of the polishing bar supporting vane and a lower end portion of the polishing bar supporting vane extending from both sides of the lower end of the motor supporting portion and separated by the opening portion included in the polishing bar through-
Wherein the upper end of the polishing-bar supporting vane and the lower end of the polishing-bar supporting vane include fastening holes that can be fastened with bolts and nuts, respectively. The method according to claim 1,
And a capillary tube is mounted at an end of the sample introduction tube. The method according to claim 1,
Wherein the sample plate manipulation unit is movable along two axes, X and Y, respectively. The method according to claim 6,
Wherein the motor supported by the motor support determines the movement of the spotting number according to the repetitive motion using the Quotient & Remainder function. Liquid chromatography (LC) capable of separating and separating a sample to be separated in a liquid-phase mixed sample into a dissolved state;
A sample introduction device of a liquid chromatography-Maldø mass spectrometer according to any one of claims 1 to 9; And
(MALDI) mass spectrometer, comprising a MALDI mass spectrometer. In a method for manufacturing a sample introduction device of a liquid chromatography (LC) -maldi mass spectrometer using a 3D printer,
a) a top end of a support base supported by a 3D printer and a bottom end of a syringe support base, wherein a central lower surface of the upper end of the syringe support includes a recess to surround the outer surface of the syringe, and a central upper surface of the syringe support base includes a syringe Manufacturing a cylinder support including a concave portion by printing a polymer material to surround the outer surface;
b) inserting a magnet on at least a lower surface of at least a portion of the upper end of the syringe support and at least a portion of the upper surface of the lower end of the syringe support so that the upper end of the syringe support and the lower end of the syringe support can be removed by the magnet;
c) manufacturing a motor support supporting a lead screw nut support and a step motor supporting a lead screw nut pushing the piston end of the syringe;
d) Combine the syringe support, the motor support supporting the leadscrew nut support and the step motor Manufacturing a syringe pump; And
e) coupling the syringe pump with an LC mass spectrometer, the LC mass spectrometer comprising: a first sample root tube connected to a syringe to introduce a MALDI mattress solution (A); A second sample root tube into which a solution (B) containing a sample separated and eluted in liquid chromatography (LC) is introduced; A mixer for mixing the solution of the first sample route tube with the solution of the second sample route tube; A sample solution for discharging the mixed solution of the MALDI mattress solution (A) and the solution (B) containing a sample eluted and separated in liquid chromatography (LC) from the mixer to a sample plate of a MALDI mass spectrometer was introduced tube; A sample plate operating part for adjusting the position of the sample plate; And a sample plate into which the mixed solution is introduced. The method comprises: preparing a liquid chromatography (LC) -maldi mass spectrometer sample introduction device using a 3D printer;

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