TWI452601B - Thin layer chromatography spray ionization device and mass spectrometer - Google Patents

Description

Thin layer chromatography electrospray ionization device and mass spectrometry system

The present invention relates to a method for analyzing a sample component, and more particularly to a thin layer chromatography electrospray free device. The invention also relates to a mass spectrometry system comprising the thin layer chromatography electrospray free device.

By mass spectrometry, one can know the molecular weight of the analytes in a sample, and then cooperate with further comparison to confirm the true identity of the analyte. Therefore, since the early 20th century, the mass spectrometry was performed. The technical mass spectrometer has become an identification tool widely used in various fields because of its advantages of easy operation and quick detection results.

The applicant has been working on the field of mass spectrometry. In 1999, Direct Electrospray Probe (DEP) was developed, which is a copper wire wound around a tipped copper coil that will be analyzed. The liquid sample is dropped directly onto the copper coil and a high voltage is applied to cause the sample to electrospray at the tip of the copper coil, thereby freeing the analyte ( Rapid Commun. Mass Spectrom. 1999 , 13 , 21-25). In the same year, the above technology was improved by arranging two fibers side by side, winding the wire at the end, and placing the liquid sample on the wire. The sample will adhere to the wire due to the surface tension of the liquid. And applying a high voltage causes the sample to be driven by the electric field force to advance through the tube formed by the two fibers, and finally an electric spray is generated at the end and the analyte is released. In 2003 and 2005, the applicant also proposed paper research related to electrospray free ( Rapid Commun. Mass Spectrom. 2003; 17: 1709-1713; and Anal. Chem. 2005, 77, 8170-8173).

In recent years, there have been papers published on the use of leaves and toothpicks as DEP probes to analyze small molecules, protein standards or small molecules of drugs in plant leaves, or to use paper as a direct electrospray free source probe. It is applied to the analysis of dry blood on the surface of paper ( Chem. Commun., 2011, 47 , 2811-2813), but paper can only be used as a carrier for sample solutions, and it is impossible to separate complex samples. To analyze a complex sample, the sample must be separated and then extracted before the free analysis can be performed. It requires complicated front-end work and is quite inconvenient to use.

Accordingly, a first object of the present invention is to provide a thin layer chromatography electrospray free device in combination with thin layer chromatography and direct electrospray free method.

Therefore, the thin layer chromatography electrospray free device of the present invention is used for separating a mixture and performing electrospray free action to form an analyte ion, and the analyte ion is further moved toward an inlet of a mass spectrometer for mass spectrometry. The thin layer chromatography electrospray free device comprises an electrode unit and a carrying unit.

The electrode unit includes a fixing base, and a voltage supply source electrically connected to the fixing base and applying a high voltage.

The carrying unit comprises a thin layer chromatography piece detachably disposed on the fixing base, and an infusion tube for providing a liquid extracting layer to the thin layer chromatography piece, wherein the thin layer chromatography piece has a tip toward the entrance of the mass spectrometer and a nip opposite the tip.

A second object of the present invention is to provide a mass spectrometry system comprising A mass spectrometer having an inlet, and a thin layer chromatography electrospray free device as described in the first object of the present invention.

After the mixture is formed into a solution, the thin layer chromatography piece is absorbed in a spotted manner to form a sample spot, and the thin layer chromatography piece is placed on the holder with the tip facing the inlet of the mass spectrometer. The infusion tube applies a flushing liquid between the sample point and the clamping portion, and applies a high voltage to the voltage supply source to move the extracting liquid toward the tip end of the thin layer chromatography sheet and drive the test. The mixture in the sample moves, and the composition of the mixture is separated according to the difference of the force with the mobile phase and the stationary phase. When the extract reaches the tip, it is affected by the high voltage electric field and is electrically sprayed at the tip, and the mixture is The component that moves the extract to the tip forms a plurality of charged analyte ions and enters the inlet of the mass spectrometer for analysis.

The utility model has the advantages that: by using the above device, the mixture can be subjected to thin layer chromatography to separate the components, and then directly electrosprayed and analyzed by the mass spectrometer, and the separated components can be directly and quickly subjected to mass spectrometry, Traditional extraction and purification steps are required to streamline the process.

The above and other technical contents, features and advantages of the present invention will be apparent from the following detailed description of the preferred embodiments.

Referring to Figure 1, a preferred embodiment of a mass spectrometry system of the present invention comprises a mass spectrometer 2 having an inlet 21 and a thin layer chromatography electrospray free device 3. The structure of the mass spectrometer 2 is capable of those of ordinary skill in the art. It is understood that the description will not be repeated, and the thin layer tomographic electrospray free device 3 comprises an electrode unit 4 and a carrying unit 5.

The electrode unit 4 includes a fixing base 41, and a voltage supply source 42 electrically connected to the fixing base 41 and applying a high voltage. The carrying unit 5 includes a thin layer chromatography piece 51 detachably disposed on the fixing base 41, and an infusion tube 52 for supplying a liquid to the thin layer chromatography piece 51, wherein the thin tube The layer chromatography sheet 51 has a tip end 511 facing the inlet 21 of the mass spectrometer 2, and a holding portion 512 opposite to the tip end 511.

In this embodiment, the fixing base 41 is exemplified by an alligator clip. The thin layer chromatogram 51 is exemplified by a shape of being cut into a triangle, but not limited thereto, as long as the tip 511 is formed. can. It should be particularly noted that the infusion tube 52 is a capillary tube and can be fixed at a predetermined position in any manner. However, the fixing method is not the focus of the present invention, and thus is not particularly depicted in the drawings. Further, in addition to the capillary tube, the infusion tube 52 may be a dropper or any means for providing a liquid through capillary action, and is not limited to the one disclosed in the embodiment.

When the mixture is to be analyzed, after the mixture is formed into a solution, the thin layer chromatography sheet 51 is sucked by a pipette (not shown) to absorb a mixture solution to form a sample point 6, and The thin layer chromatography piece 51 is disposed on the fixing base 41, and the tip end 511 faces the inlet 21 of the mass spectrometer 2. The method of spotting the suction device is understood by those having ordinary knowledge in the field of thin layer chromatography. ,No longer.

The infusion tube 52 applies an elution (or mobile phase) between the sample point 6 and the clamping portion 512, and applies a high voltage to the voltage supply source 42. Electric, moving the liquid toward the tip 511 and driving the mixture in the sample point 6 to move, so that the composition of the mixture is separated according to the difference between the moving phase and the stationary phase, and the force of the stationary phase The smaller the force and the greater the force with the mobile phase, the faster the movement speed. When the extract reaches the tip 511, it is subjected to a high-voltage electric field and is electrically sprayed off at the tip 511 to form a charged droplet, and the composition of the mixture that moves to the tip 511 as the extract moves is merged with the charged droplet. A plurality of charged analyte ions 7 are formed and enter the mass spectrometer 2 via the inlet 21, and analyzed by the mass spectrometer 2 to generate a mass spectrogram.

2 and 3 are experimental examples of testing by the mass spectrometry system of the present invention for analyzing whether or not the blood contains methyl amphetamine. A blood sample is taken and analyzed by the mass spectrometer 2 after the steps of spotting, separating, electrospraying, etc. by the mass spectrometry system. Figure 2 (A) is the total ion chromatogram (TIC) of the blood sample, Figure 2 (B) is the ion chromatogram of methyl amphetamine (EIC of m / z 150), Figure 3 is the methyl amphetamine Mass spectrum. It should be noted that when a blood sample is moved by the extract liquid on the thin layer chromatography sheet 51, macromolecules such as blood cells and proteins in the blood sample can be retained in the thin layer chromatography because the moving speed is slow. The side of the sheet 51 close to the clamping portion 512 is such that the analysis result of the small molecules in the blood sample is not interfered by the signals of macromolecules such as blood cells and proteins, and the matrix interference effect from the whole blood can be effectively reduced.

In addition, it should be particularly noted that the thin layer chromatography sheet 51 has a triangle which is tapered by the clamping portion 512 toward the tip end 511 as shown in FIG. 1, or may be as shown in FIG. Having a tip 511 to the The hem 513 extends in the direction of the clamping portion 512 and is bent, and the hem 513 is pleated at 180 degrees. The reason why the hem 513 is formed is that, after the thin layer chromatography piece 51 is cut, although the cut edge is straight from the naked eye, it is possible to generate a plurality of sharp-cone structures under the microscopic shape. The shape of the conical structure also causes the phenomenon of electrospraying to be released, which affects the intensity of the analysis result of the mass spectrometer 2. Therefore, the formation of the hem 513 allows the electrospray to concentrate on the tip 511. Of course, as shown in FIG. 5, both sides of the thin layer chromatography sheet 51 are formed with hem 513, and the pleats of the hem 513 are about 90 degrees. It should be noted that the pleat angle of the hem 513 may vary depending on actual needs, and is not limited to those disclosed in the embodiment. Further, as shown in FIG. 6, the thin layer chromatography piece 51 may have a shape in which the holding portion 512 is equal in width and is tapered from the end of the holding portion 512 toward the tip end 511. With this design, when the liquid is moved in the direction of the tip end 511, the flow rate is not easily affected by the change in the area of the holding portion 512, and the flow rate of the liquid is more even and stable.

In summary, the present invention utilizes the above-described apparatus and operation steps, and allows the mixture to be subjected to thin layer chromatography to separate the components, and then directly electrosprayed and analyzed by the mass spectrometer 2, and the separated components can be directly and quickly separated. By performing the analysis, it is not necessary to carry out the conventional extraction and purification steps, and the operation flow is simplified, so that the object of the present invention can be achieved.

The above is only the preferred embodiment of the present invention, and the scope of the invention is not limited thereto, that is, the simple equivalent changes and modifications made by the scope of the invention and the description of the invention are All remain within the scope of the invention patent.

2‧‧‧Mass Spectrometer

21‧‧‧ entrance

3‧‧‧Thin layer chromatography electrospray free device

4‧‧‧Electrode unit

41‧‧‧ Fixed seat

42‧‧‧Voltage supply

5‧‧‧ Carrying unit

51‧‧‧Thin-layer chromatography

511‧‧‧ cutting-edge

512‧‧‧ gripping department

513‧‧‧Frills

52‧‧‧Infusion tube

6‧‧‧sample points

7‧‧‧Test object ions

1 is a schematic view showing a preferred embodiment of the mass spectrometry system of the present invention; FIG. 2 is an ion chromatogram illustrating the analysis results of the experimental example of the present invention; and FIG. 3 is a mass spectrum diagram, and the experimental example of the present invention is also illustrated. Figure 4 is a schematic view showing a thin layer of chromatograms having a hem appearance in the preferred embodiment; and Figure 5 is a schematic view showing the thin layer in the preferred embodiment The strip has two hem patterns; and Fig. 6 is a schematic view showing another aspect of the thin layer chromatogram in the preferred embodiment.

2‧‧‧Mass Spectrometer

21‧‧‧ entrance

3‧‧‧Thin layer chromatography electrospray free device

4‧‧‧Electrode unit

41‧‧‧ Fixed seat

42‧‧‧Voltage supply

5‧‧‧ Carrying unit

51‧‧‧Thin-layer chromatography

511‧‧‧ cutting-edge

512‧‧‧ gripping department

52‧‧‧Infusion tube

6‧‧‧sample points

7‧‧‧Test object ions

Claims (8)

A thin layer chromatography electrospray free device is used for separating a mixture and performing electrospray free action, and then moving toward an inlet of a mass spectrometer for mass spectrometry. The thin layer chromatography electrospray free device comprises: The electrode unit includes a fixing base, and a voltage supply source electrically connected to the fixing base and applying a high voltage; and a carrying unit comprising a thin layer chromatography piece detachably disposed on the fixing base, And an infusion tube for providing an extract to the thin layer chromatography sheet, wherein the thin layer chromatography sheet has a tip end facing the inlet of the mass spectrometer, and a clamping portion opposite to the tip end; After the mixture is formed into a solution, the thin layer chromatography piece is absorbed in a spotted manner to form a sample point, and the thin layer chromatography piece is placed on the fixing seat, and the tip end faces the inlet of the mass spectrometer, and the infusion tube is A liquid is applied between the sample point and the clamping portion, and the voltage supply source is applied with high voltage, and the liquid is moved toward the tip end of the thin layer chromatography sheet to drive the sample point. The mixture moves to separate the components of the mixture, When the extract reaches the tip of the thin layer chromatography, it is electrically sprayed at the tip by the high voltage electric field, and a plurality of charged analytes are formed in the mixture as the extract moves to the tip. Ions enter the inlet of the mass spectrometer for analysis. The thin layer chromatography electrospray free apparatus according to claim 1, wherein the thin layer chromatography sheet further has at least one hem extending from the tip end toward the nip portion and being bent and formed. Thin layer chromatography electrospray free loading according to item 1 of the patent application scope And wherein the thin layer chromatography piece is a triangle which is tapered by the clamping portion toward the tip end. The thin layer chromatography electrospray free apparatus according to claim 1, wherein the nip portion of the thin layer chromatography sheet is of equal width and is tapered from the end of the nip portion toward the tip end. . A mass spectrometry system for separating a mixture and performing mass spectrometry, the mass spectrometry system comprising: a mass spectrometer having an inlet; and a thin layer chromatography electrospray ionization apparatus comprising: an electrode unit comprising a fixing seat, And a voltage supply source electrically connected to the fixing base and applying a high voltage; and a carrying unit comprising a thin layer chromatography piece detachably disposed on the fixing base, and a thin layer for the thin layer The chromatogram provides an infusion tube for the extract, wherein the thin layer chromatography has a tip toward the inlet of the mass spectrometer and a nip opposite the tip; the mixture forms a solution and is spotted The thin layer chromatography piece is absorbed to form a sample point, and the thin layer chromatography piece is disposed on the fixing seat, the tip end faces the inlet of the mass spectrometer, and the infusion tube is at the sample point and the clamping part Applying a liquid between the voltage supply and applying the high voltage to the voltage supply, moving the liquid toward the tip end of the thin layer chromatography and driving the mixture in the sample point to move the composition of the mixture Separation, when the extract reaches the thin layer chromatography When the tip is influenced by high-voltage electric field is generated in the sprayed free tip, and the mixture as an eluent movement The components to the tip form a plurality of charged analyte ions and enter the inlet of the mass spectrometer for analysis. The mass spectrometry system of claim 5, wherein the thin layer chromatography sheet further has at least one hem extending from the tip end toward the nip portion and being bent and formed. The mass spectrometry system according to claim 5, wherein the thin layer chromatography sheet has a triangular shape which is tapered from the nip portion toward the tip end. The mass spectrometry system of claim 5, wherein the nip portion of the thin layer chromatography sheet is of equal width and tapers in the direction of the tip end.