CN110858532A - Sample introduction device, sample introduction system and sample introduction method of mass spectrometer - Google Patents
Sample introduction device, sample introduction system and sample introduction method of mass spectrometer Download PDFInfo
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- CN110858532A CN110858532A CN201810967541.XA CN201810967541A CN110858532A CN 110858532 A CN110858532 A CN 110858532A CN 201810967541 A CN201810967541 A CN 201810967541A CN 110858532 A CN110858532 A CN 110858532A
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/02—Details
- H01J49/04—Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components
- H01J49/0431—Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components for liquid samples
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/26—Mass spectrometers or separator tubes
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Abstract
The invention provides a sample introduction device of a mass spectrometer, which comprises a carrier, wherein the carrier comprises a bearing disc and at least one nozzle structure, the nozzle structure is made of a conductive material, the nozzle structure is arranged on the bearing disc, the nozzle structure is provided with a nozzle, a sample groove is formed in the nozzle structure, the sample groove is communicated with the nozzle, and a gas channel communicated with two surfaces of the nozzle structure is arranged at a position, close to the nozzle, of the nozzle structure. When the ion source type micro-droplet analyzer is used, a sample to be detected is injected into the sample tank, gas is provided by the gas supply device and passes through the gas channel to form negative pressure at the nozzle, so that the sample to be detected in the sample tank is atomized into micro-droplets and is sprayed out from the nozzle, and meanwhile, a high voltage difference is formed between the nozzle structure and the free substance injection port through the ion source, so that analyte molecules existing in the micro-droplets form ions when leaving the nozzle structure, and the analyte ions enter the analysis device to perform mass spectrometry.
Description
Technical Field
The invention relates to the field of mass spectrometers, in particular to a sample introduction device, a sample introduction system and a sample introduction method of a mass spectrometer.
Background
The mass spectrometer mainly comprises a sample introduction device, an ion source, a mass analyzer and a detector, wherein the sample introduction device introduces a sample to be detected into the ion source, the sample to be detected is ionized in the ion source to generate ions, the ions enter the mass analyzer under the action of an accelerating electric field, the mass analyzer separates the ions with different charge-mass ratios, and the separated ions enter the detector to obtain a mass spectrogram, so that the molecular weight of target molecules in the sample to be detected is analyzed.
When the sample to be detected is liquid, the sample feeding device may include a pump and a capillary, and the liquid sample is pushed by the pump to be introduced into the ion source through the capillary. The sample introduction device of the conventional mass spectrometer usually has a capillary with a long path (for example, tens of centimeters) to form a sample introduction channel. Since the inner diameter of the capillary is small (e.g., less than 150 μm), the liquid sample is pre-treated to remove impurities, which would otherwise cause clogging of the capillary, however, the pre-treatment step is complicated to affect the analysis efficiency, and the volume of the liquid sample is required to be large (e.g., more than 1mL), resulting in limitation on analysis of a trace amount of the liquid sample. Furthermore, the cleaning of the capillary tubing after use is also time consuming and consumes a large amount of solvent.
Disclosure of Invention
In view of the above situation, the present invention provides a sample introduction device, a sample introduction system, and a sample introduction method for a mass spectrometer, which are suitable for analyzing a trace amount of liquid sample.
The sample introduction device comprises a carrier, wherein the carrier comprises a bearing disc and at least one nozzle structure, the nozzle structure is made of conductive materials, the nozzle structure is arranged on the bearing disc, the nozzle structure is provided with a nozzle, a sample groove is formed inside the nozzle structure, the sample groove is communicated with the nozzle, a gas channel communicated with two sides of the nozzle structure is arranged at a position, close to the nozzle, of the nozzle structure, and the gas channel is provided with a gas inlet and a gas outlet.
The sample introduction system comprises a sample introduction hole arranged on an ion source and the sample introduction device, wherein a carrier of the sample introduction device is inserted into the sample introduction hole.
The sample introduction method is applied to the sample introduction device, and comprises the following steps: injecting a sample to be detected into a sample groove; target molecules in the sample to be detected react and combine with the functional groups of the functional group coating, and non-target molecules in the sample to be detected flow out of the nozzle; injecting a washing buffer solution into the sample tank; a cleaning solution is injected into the sample cell.
The sample introduction device, the sample introduction system and the sample introduction method of the mass spectrometer provide that the liquid sample can be directly injected into the sample groove, and the carrier is inserted into the sample introduction hole of the ion source to complete the sample introduction step, so that the operation is easy, the analysis efficiency is improved, a large amount of liquid samples are not required to be pretreated, and the sample introduction device, the sample introduction system and the sample introduction method are suitable for analyzing trace liquid samples.
Drawings
Fig. 1 is a schematic perspective view of a sample injection device according to an embodiment of the present invention.
FIG. 2 is a schematic cross-sectional view of a nozzle configuration according to an embodiment of the present invention.
Fig. 3 and 4 are schematic sectional views of a nozzle structure according to an embodiment of the present invention in a use state.
Fig. 5 is a schematic side view of the sample injection device according to an embodiment of the present invention.
Fig. 6 is a schematic perspective external view of a sample injection device according to another embodiment of the present invention.
FIG. 7 is a schematic cross-sectional view of a nozzle configuration according to another embodiment of the present invention.
Fig. 8A to 8D are schematic cross-sectional views illustrating a use state of a nozzle structure according to another embodiment of the present invention.
Fig. 9 is a flow chart of a sample injection method of a sample injection device according to another embodiment of the invention.
FIG. 10 is a schematic cross-sectional view of a nozzle configuration according to another embodiment of the present invention.
Fig. 11 and 12 are mass spectra of samples analyzed using a nozzle structure according to another embodiment of the present invention.
Description of the main elements
Detailed Description
The following detailed description will further illustrate the invention in conjunction with the above-described figures.
As shown in fig. 1, a sample introduction device 100 of a mass spectrometer of the present invention includes a carrier 1 and a hand-held plate 2.
The carrier 1 comprises a carrier tray 11 and at least one nozzle structure 12, the nozzle structure 12 is made of a conductive material (such as metal), the nozzle structure 12 is arranged on the carrier tray 11, as shown in fig. 2, the nozzle structure 12 is provided with a spout 121, a sample groove 122 is formed inside the nozzle structure 12, the sample groove 122 is funnel-shaped, the sample groove 122 is communicated with the spout 121, the notch of the sample groove 122 is larger than the spout 121, the notch of the sample groove 122 is larger so as to facilitate the injection of a liquid sample, and the spout 121 is smaller so as to enable the liquid sample to be free from external force and from downward leakage under the action of surface tension of the liquid sample. The nozzle structure 12 is provided with a gas channel 123 adjacent to the nozzle opening 121 and communicating the top surface and the bottom surface of the nozzle structure 12, the gas channel 123 is annular and surrounds the side surface of the sample chamber 122, the gas channel 123 has a gas inlet 1231 and a gas outlet 1232, and the gas outlet 1232 is annular and surrounds the nozzle opening 121. The present embodiment is illustrated by using the annular gas channel 123, but the present invention is not limited thereto, and persons skilled in the art can substitute one-sided gas channel, semi-annular gas channel, or multiple elongated gas channels. The number of nozzle structures 12 may be set as desired, and the top surface of each nozzle structure 12 may be provided with an identification feature 124, such as a one-dimensional barcode, a two-dimensional barcode or an identification circuit, for use in mass spectrometry identification to identify the sample 200 to be tested and its target molecules in the sample well 122 of the nozzle structure 12 currently being processed, and the ion source 300 may adjust control parameters, such as voltage or gas pressure, accordingly. In another embodiment, the sample slot 122b is used to carry a functional group coating 125b (as shown in fig. 7) attached to the surface of the sample 200 to be tested, and the functional group of the functional group coating 125b can react and combine with the target molecule in the sample 200 to be tested, so as to achieve the purposes of cleaning and concentrating the analyte, and thus the sensitivity and accuracy of mass spectrometry can be improved.
The hand-held plate 2 is fixed on one side of the carrier 1 so as to be convenient for an analyst to hold and operate, and the hand-held plate 2 and the carrier plate 11 are made of an insulating material (e.g., plastic).
In practical use, as shown in fig. 3, the sample introduction device 100 of the present invention firstly injects a sample 200 to be measured into the sample tank 122, as shown in fig. 5, then, the carrier 1 is inserted into the ion source 300 through the sample inlet 301, and a high voltage is applied by the ion source 300 to form a high voltage difference between the nozzle structure 12 and the free substance inlet 400, so that ions are formed when the sample 200 to be tested in the sample tank 122 leaves the nozzle structure 12 as shown in fig. 4, while gas is supplied through the gas passage 123 through the gas inlet 1231 by the gas supply means 302, so that a negative pressure is formed at the nozzle 121 and the gas exiting from the gas outlet 1232 accelerates the ions to exit the nozzle structure 12, and, in addition, ions exiting from the nozzle structure 12 are drawn into the free species injection port 400 for analysis by creating a pressure differential between atmospheric pressure within the ion source 300 and vacuum pressure within the analysis apparatus 500.
The high voltage difference between the nozzle structure 12 and the free substance injection port 400 is about 4.5-5.5 kV, but the actual voltage difference is adjusted according to the distance between the nozzle structure 12 and the free substance injection port 400. The distance between the nozzle structure 12 and the free substance injection port 400 is about 0.5-2 cm, preferably 1-1.5 cm. The ion source 300 is provided with one or more sample inlets 301 on the front or side thereof.
In one embodiment, when the sample 200 to be tested forms positive ions upon exiting the nozzle structure 12, the nozzle structure 12 is connected to ground potential and the ion source 300 applies a negative high voltage to the dissociated species injection port 400, or alternatively, the dissociated species injection port 400 is connected to ground potential and the ion source 300 applies a positive high voltage to the nozzle structure 12.
The sample introduction device 100 provided by the invention has the advantages that the sample 200 to be detected can be directly injected into the sample groove 122, the carrier 1 is inserted into the sample introduction hole 301 of the ion source 300, the sample introduction step is completed, the operation is easy, the analysis efficiency is improved, the sample 200 to be detected only needs to be taken by 1-5 mu L, the pretreatment of a large amount of samples 200 to be detected is not needed, and the sample introduction device is suitable for analyzing trace liquid samples. Since the volume of the sample cell 122 is fixed, mass spectrometry can be performed not only for qualitative analysis but also for quantitative analysis.
Referring to fig. 1 to 5, a sample injection device 100 according to an embodiment of the present invention is shown, wherein a nozzle structure 12 is disposed on a carrier tray 11.
Fig. 6 shows a sample injection device 100a according to another embodiment of the present invention, wherein a plurality of nozzle structures 12a are arranged on a carrier tray 11a at intervals. In the present embodiment, six nozzle structures 12a are provided at intervals on the carrier tray 11 a. An automated device (not shown) may be disposed within the ion source 300 to sequentially inject different samples 200 to be tested into the sample wells 122a of the plurality of nozzle structures 12 a. A scanning device 303 (shown in fig. 5) may also be disposed in the ion source 300, and may store information of the sample 200 to be measured in the sample slot 122a of each nozzle structure 12a according to the identification feature 124a of each nozzle structure 12a and transmit the information to the mass spectrometer, or store information of the sample 200 to be measured in the sample slot 122a of each nozzle structure 12a through a storage device (not shown) disposed on the carrier tray 11a, and read the stored information through a reading device (not shown) when the carrier 1a is inserted into the ion source 300.
Referring to fig. 7 to 8D, which illustrate a nozzle structure 12b according to another embodiment of the present invention, a different functional group coating 125b is attached to the surface of the sample groove 122b of the nozzle structure 12b, the functional group coating 125b may include nanoparticles, and the functional group coating 125b may be a hydrophobic C8 or C18 alkyl coating, a hydrophilic nitrile (nitrile) or amide (amide) coating, or other coating having intermolecular forces. The functional groups of the coating 125b can react and combine with the target molecules 201b in different samples 200b to be tested, so as to improve the sensitivity and accuracy of mass spectrometry.
In another embodiment, the surface of the sample well 122b of the nozzle structure 12b is adhered with a silicon-containing coating (not shown), which may comprise a glass, quartz, or silica gel composition. To analyze the sample 200b, a solution containing functional groups is first injected into the sample chamber 122b, and after the functional groups react with silicon atoms and are bonded for a period of time, the solution is removed. For example, when the target molecule 201b in the sample 200b to be analyzed is protein and the C18 alkyl coating is needed, the solution containing the C18 alkyl is injected into the sample container 122b, and after adding the appropriate reagent and controlling the ph, the C18 alkyl is bonded to the silicon atom, and the solution is removed, so that the C18 alkyl coating is attached to the sample container 122 b.
Please refer to fig. 8A to 8D, which are schematic diagrams illustrating steps of a sample injection method according to the present invention. In one embodiment, the steps of fig. 8A-8D are performed by automated equipment within the ion source 300, in another embodiment, the steps of fig. 8A are performed manually outside of the ion source 300, and the steps of fig. 8B-8D are performed by automated equipment within the ion source 300. As shown in FIG. 9, the sample injection method is as follows.
Step 901: as shown in fig. 8A, a sample 200b to be measured is injected into the sample well 122 b.
Step 902: as shown in fig. 8B, the target molecules 201B in the sample to be tested 200B are reacted and bonded with the functional groups of the functional group coating 125B, and the non-target molecules 202B in the sample to be tested 200B slowly flow out from the nozzle 121B. After a predetermined reaction time, gas is supplied through the gas channel 123b by the gas supply device 302, so that a negative pressure is formed at the jet 121b to accelerate the non-target molecules 202b out of the jet 121 b.
Step 903: as shown in fig. 8C, a wash buffer 600b is injected into the sample well 122b to wash away residual non-target molecules 202 b. Gas is supplied through the gas channel 123b by the gas supply 302 so that a negative pressure is created at the jet 121b to carry the remaining non-target molecules 202b out of the jet 121 b.
Step 904: as shown in fig. 8D, a cleaning solution (solution) 700b is injected into the sample well 122b to break the force between the target molecule 201b and the functional group of the functional group coating 125 b. The target molecules 201b are ionized by applying a high voltage to the ion source 300 to form a high voltage difference between the nozzle structure 12b and the free substance inlet 400, and the gas is supplied through the gas channel 123b by the gas supply device 302, so that a negative pressure is formed at the nozzle 121b to send the free molecules with the target molecules 201b to the analysis device 500, and the analysis device 500 performs mass spectrometry, and the related operations can be described with reference to fig. 4.
For example, when it is desired to confirm whether the sample 200b to be tested contains protein, the functional group coating 125b is a C18 alkyl coating. In the step of fig. 8B, if the sample 200B contains proteins, the proteins and the C18 alkyl groups are bonded by intermolecular forces, and after the remaining sample 200B flows out from the nozzle 121B, inorganic or organic salts may be attached to the surface of the sample well 122B, so that in the step of fig. 8C, unnecessary salts are washed away by using the washing buffer 600B to reduce interference of the salts during mass spectrometry. In the step of fig. 8D, a suitable wash 700b is used to break the forces between the protein and the C18 alkyl groups.
As shown in fig. 10, a nozzle structure 101 according to another embodiment of the present invention is provided, the nozzle structure 101 includes a base 102, a funnel 103 and an air supply pipe 104, the base 102 has a top plate 1021 and a ring side wall 1022 connected to a bottom surface of the top plate 1021, the ring side wall 1022 is tapered and has an air outlet channel 1023, a gas chamber 1024 is formed between the top plate 1021 and the ring side wall 1022, the gas chamber 1024 is communicated with the air outlet channel 1023, a tube 1032 is connected to a bottom end of the funnel 103 having a carrier 1031 and the carrier 1031, a sample slot 1033 is formed inside the carrier 1031, a nozzle 1034 is formed at a bottom end of the tube 1032, the funnel 103 penetrates through the top plate 1021 of the base 102 through the tube 1032, the tube 1032 is located in the gas chamber 1024, the air outlet channel 1023 is annular and surrounds the tube 1032, the air supply pipe 104 penetrates through the top plate 1021 of the base. The annular side walls 1022 and funnel 103 are made of a conductive material.
In practical use, the mass spectrometry scan parameters are set, the voltage is set to 5.5kV, the gas pressure is set to 20psi, the volume is 100 μ L, and the concentration is 1.0 × 10-4The sample to be measured of M is injected into the sample tank 1033, ions are generated in the ion source 300 by the sample to be measured, and the ions are sent to the analyzing apparatus 500, thereby obtaining a mass spectrum.
When the sample to be detected is micromolecule Rhodamine 6G (Rhodamine 6G), setting the mass spectrum scanning range to be between m/z 50-500, and obtaining a mass spectrum as shown in figure 11, wherein the signal of the detected Rhodamine 6G ion is m/z 443.3.
When the sample to be detected is macromolecular cytochrome C (cytochrome C), setting the mass spectrum scanning range to be m/z 600-1500, and obtaining a mass spectrum as shown in figure 12, wherein the obtained mass spectrum shows that the measured ion signal distribution is m/z 782.1, m/z 817.4, m/z 874.5, m/z 941.9, m/z 1020.2 and m/z 1113.0, which is the protein multivalent charge ion distribution presented by cytochrome C molecules in mass spectrum analysis.
However, the above description is only a preferred embodiment of the present invention, and should not be taken as limiting the scope of the invention, which is defined by the appended claims and the description of the invention. Furthermore, it is not necessary for any embodiment or claim of the invention to address all of the objects, advantages, or features disclosed herein. In addition, the abstract and the title of the invention are provided for assisting the search of patent documents and are not intended to limit the scope of the invention.
Claims (15)
1. The utility model provides a sampling device, its characterized in that, includes the carrier, the carrier is including bearing dish and at least one nozzle structure, nozzle structure is made by conducting material, nozzle structure sets up bear on the dish, nozzle structure has the spout, nozzle structure's inside forms the sample cell, the sample cell with the spout intercommunication, nozzle structure is close to spout department is equipped with the intercommunication the gas passage on nozzle structure two sides, gas passage has air inlet and gas outlet.
2. The sample introduction device according to claim 1, wherein the sample well is funnel-shaped.
3. The sample introduction device according to claim 1, wherein the gas channel is annular and surrounds the side surface of the sample cell.
4. The sample introduction device according to claim 3, wherein the gas outlet of the gas channel is annular and surrounds the nozzle.
5. The sample introduction device according to claim 1, wherein the nozzle structure is provided with an identification feature.
6. The sample introduction device according to any of claims 1 to 5, further comprising a hand-held plate fixed to one side of the carrier.
7. The sample introduction device according to claim 6, wherein a plurality of the nozzle structures are arranged on the carrier tray at intervals.
8. The sample introduction device according to any of claims 1 to 5, wherein a functional group coating is attached to the surface of the sample well.
9. The sample introduction device according to claim 8, wherein the functional group coating comprises nanoparticles.
10. A sample introduction system comprising a sample introduction well provided on an ion source and a sample introduction device according to any of claims 1 to 9, the carrier of the sample introduction device being inserted into the sample introduction well.
11. The sample introduction system according to claim 10, further comprising a gas supply device that provides gas through the gas channel.
12. The sample introduction system according to claim 10, further comprising a scanning device that stores information of a sample to be measured in the sample well of the nozzle structure according to the identification feature of the nozzle structure.
13. The sample introduction system according to claim 12, wherein the ion source adjusts a control parameter according to information of the sample to be measured.
14. The sample introduction system according to any of claims 10 to 13, wherein the ion source forms a high voltage difference between the nozzle structure and a free species injection port.
15. A sample introduction method, applied to the sample introduction apparatus according to claim 8 or 9, the method comprising:
injecting a sample to be tested into the sample groove;
target molecules in the sample to be detected are combined with the functional groups of the functional group coating in a reaction way, and non-target molecules in the sample to be detected flow out of the nozzle;
injecting a wash buffer into the sample cell;
injecting a cleaning solution into the sample cell.
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| CN201810967541.XA CN110858532A (en) | 2018-08-23 | 2018-08-23 | Sample introduction device, sample introduction system and sample introduction method of mass spectrometer |
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| CN201810967541.XA CN110858532A (en) | 2018-08-23 | 2018-08-23 | Sample introduction device, sample introduction system and sample introduction method of mass spectrometer |
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| US20040229377A1 (en) * | 2003-04-14 | 2004-11-18 | National Cheng Kung University | Microfluidic chip system integrated with nano-electrospray interface and method using thereof |
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