Instrument and method for integrating extreme ultraviolet dissociation spectrum-mass spectrum with spectroscopy

Abstract

The invention relates to an extreme ultraviolet dissociation spectrum-mass spectrum integrated spectroscopy instrument and method, a mass spectrum system is provided with a through hole, the through hole is provided with a first on-off control valve, a vacuum connection pipeline system comprises a five-way joint, and the five-way joint is provided with a second on-off control valve. Interfaces of the five-way joint are respectively connected with a joint assembly, a second on-off control valve, an observation window, a laser high-transmittance window and a reflector system, the lower end of the joint assembly is connected with a vacuum pump through a pipeline, and the second on-off control valve is connected with an ultraviolet laser pipeline; the laser is transmitted through the laser transmission pipeline and then enters from the laser high-transmittance window, and a reflector in the reflector system is adjustably arranged in the five-way connector. According to the invention, vacuum ultraviolet laser and atmospheric pressure ultraviolet laser can be coupled in a non-stop state of a mass spectrum, and wavelength-adjustable extreme ultraviolet laser can be introduced into a mass spectrum dissociation ion trap to carry out ultrafast photodissociation on biomolecules such as protein and the like, so that a multi-dimensional spectroscopy technology is realized.

Description

Technical Field

[0001] The invention relates to the technical field of spectrum and mass spectrum analysis, in particular to an instrument and method for extreme ultraviolet light dissociation spectrum-mass spectrum integrated spectroscopy. Background Art

[0002] With the rapid development of mass spectrometry technology, mass spectrometry has become a key analytical tool for analyzing proteins and metabolites. The analysis of samples by tandem mass spectrometry often includes one or more ion dissociation stages, which obtain product ions by dissociating ions generated by the sample, and measure the intensity and mass-to-charge ratio of the product ions to clarify or quantify target or non-target analytes with high specificity. The current methods for obtaining product ions mainly include collision-induced dissociation (CID), collision-activated dissociation (CAD), electron transfer dissociation (ETD) and ultraviolet photodissociation (UVPD).

[0003] Ultraviolet photodissociation (UVPD) is a basic process that uses ultraviolet laser light emitted by an ultraviolet laser to irradiate precursor ion groups to fragment them and generate product ions. This technology can promote more extensive and in-depth fragmentation of proteins, can provide a high sequence detection range, and is very suitable for high-throughput proteomics. However, the current ultraviolet photodissociation (UVPD) technology is limited to certain specific atmospheric pressure ultraviolet laser bands, which cannot continuously cover the vacuum ultraviolet band, and cannot provide spectra with both sample spectral information and mass spectral information, which seriously limits the development of ultraviolet photodissociation (UVPD) technology. Summary of the invention

[0004] The purpose of the present invention is to provide an instrument and method for extreme ultraviolet photodissociation action spectrum mass spectrometry integrated spectroscopy, which can couple the functions of vacuum ultraviolet laser and atmospheric pressure ultraviolet laser without stopping the mass spectrometer through the joint action and conversion of a first on-off control valve, a second on-off control valve, a reflector adjustment system and an adjustment platform system, and can introduce 10nm-180nm adjustable wavelength extreme ultraviolet laser into the mass spectrometry dissociation ion trap to achieve ultrafast photodissociation of biological molecules such as proteins by ultraviolet lasers of different wavelengths.

[0005] The objective of the present invention is achieved through the following technical solutions:

[0006] An instrument for extreme ultraviolet light dissociation action spectroscopy mass spectrometry integrated spectroscopy, comprising a mass spectrometry system, wherein a mass spectrometry dissociation ion trap is arranged inside the mass spectrometry system, and further comprising an adjustment platform system, a laser system, a vacuum connection pipeline system and a reflector adjustment system, wherein the mass spectrometry system and the laser system are both arranged at the upper end of the adjustment platform system, a through hole coaxial with the center of the mass spectrometry dissociation ion trap annular electrode is arranged on a vacuum sealing plate of the mass spectrometry system, and a first on-off control valve is arranged on the through hole, the vacuum connection pipeline system comprises a five-way joint, and a first interface of the five-way joint is connected to the first on-off control valve through a joint assembly, the lower end of the joint assembly is connected to a vacuum pump through a pipeline, and the five-way joint is provided with a first on-off control valve. The second interface is connected to the ultraviolet laser pipeline through the second on-off control valve, the third interface of the five-way joint is provided with an observation window, the fourth interface of the five-way joint is provided with a laser high-transmittance window, and the laser emitted by the laser system is reflected and transmitted by a transmission reflector on a laser transmission support pipeline and then shot into the five-way joint through the laser high-transmittance window. The reflector adjustment system includes an adjusting reflector and a reflector adjustment mechanism. The adjusting reflector is arranged inside the five-way joint, and the laser shot into the five-way joint is reflected by the adjusting reflector and shot into the mass spectrometry system. The reflector adjustment mechanism is installed on the fifth interface of the five-way joint, and the adjusting reflector adjusts its posture through the reflector adjustment mechanism.

[0007] The joint assembly includes a connecting bellows, a three-way joint A, a three-way joint B and a vacuum gauge, wherein the first interface of the three-way joint A is connected to the first on-off control valve through the connecting bellows, the second interface of the three-way joint A is connected to the first interface of the five-way joint, the third interface at the lower end of the three-way joint A is connected to the first interface of the three-way joint B, the second interface of the three-way joint B is connected to the vacuum gauge, and the third interface of the three-way joint B is connected to the vacuum pump through a pipeline.

[0008] The reflector adjustment mechanism includes a moving device, a rotating device, a supporting bellows and a connecting rod, wherein the rotating device is driven to move by the moving device, the lower end of the supporting bellows is sleeved on the output shaft of the rotating device and is fixedly connected to the rotating device housing, the upper end of the supporting bellows is fixedly connected to the fifth interface of the five-way joint, the lower end of the connecting rod extends into the supporting bellows and is fixedly connected to the output shaft of the rotating device, and the upper end of the connecting rod is fixedly connected to the adjusting reflector.

[0009] The adjustment platform system includes a translation adjustment mechanism, a lifting electric cylinder, a lifting plate, a rotation drive device and a mass spectrometry system support plate, wherein each lifting electric cylinder is arranged on the translation adjustment mechanism, each corner end of the lifting plate is respectively hinged to the upper end of the corresponding lifting electric cylinder, the rotation drive device is arranged on the lifting plate, the mass spectrometry system support plate is arranged on the rotation drive device, and the mass spectrometry system support plate is driven to rotate by the rotation drive device, the mass spectrometry system is arranged on the mass spectrometry system support plate, and the laser system is arranged on one side of the lifting plate.

[0010] A method for an instrument using the extreme ultraviolet light dissociation action spectrum mass spectrometry integrated spectroscopy, specifically: the first on-off control valve and the second on-off control valve are closed and the adjustment reflector is not raised, the vacuum pump is first turned on to evacuate the vacuum connecting pipeline system, when the vacuum degree inside the vacuum connecting pipeline system reaches the requirement, the second on-off control valve is opened, the vacuum ultraviolet laser is injected into the five-way joint along the ultraviolet laser pipeline, and then the first on-off control valve on the mass spectrometry system is opened, and the lifting electric cylinder in the platform system is adjusted to fine-tune the inclination angle of the lifting plate, and the rotary drive device fine-tunes the horizontal angle of the mass spectrometry system so that the vacuum ultraviolet laser enters the mass spectrometry system and accurately acts on the biological sample to be dissociated.

[0011] A method for an instrument using the extreme ultraviolet light dissociation action spectrum mass spectrometry integrated spectroscopy, specifically: the first on-off control valve and the second on-off control valve are closed, and the adjusting reflector is driven to rise and adjust the angle by the reflector adjustment mechanism, and then the vacuum pump is turned on to evacuate the vacuum connection pipeline system. When the vacuum degree inside the vacuum connection pipeline system reaches the requirement, the first on-off control valve on the mass spectrometry system is opened, and the ultraviolet laser emitted by the laser system is reflected and transmitted by the transmission reflector on the laser transmission support pipeline and then shot onto the adjusting reflector through the laser high-transmittance window at the fourth interface of the five-way joint, and then enters the mass spectrometry system after being reflected by the adjusting reflector, the lifting electric cylinder in the platform system is adjusted to fine-tune the inclination angle of the lifting plate, and the rotary drive device fine-tunes the horizontal angle of the mass spectrometry system so that the vacuum ultraviolet laser enters the mass spectrometry system and accurately acts on the biological sample to be dissociated.

[0012] The ultraviolet laser pipeline is connected to the second laser system, and through the time pulse delay setting, another laser beam with a different wavelength emitted by the second laser system and the laser beam emitted by the laser system are alternately introduced into the mass spectrometry system respectively, and the adjusting reflector cooperates to adjust the angle so that the laser can accurately act on the biological sample to be dissociated.

[0013] An application of an instrument for integrating spectroscopy based on the extreme ultraviolet photodissociation action spectrum and mass spectrometry, specifically comprising: introducing a biological sample into the mass spectrometry system from a sample inlet of the mass spectrometry system, selecting an extreme ultraviolet laser with a specific wavelength in the 10nm-180nm band to irradiate the biological sample in the mass spectrometry dissociation ion trap through the vacuum connection pipeline system, and detecting the products of extreme ultraviolet photodissociation and ionization by mass spectrometry; scanning different laser wavelengths in the 10nm-180nm band, and recording the products of photodissociation and ionization at different wavelengths by mass spectrometry, to obtain multidimensional spectral information covering both extreme ultraviolet photodissociation spectrum information of biological molecules and mass spectrum information of fragment ions, and realizing single pulse or multiple pulse photodissociation of protein and / or ester biological molecules in the biological sample by the 10nm-180nm adjustable wavelength extreme ultraviolet laser, wherein the duration of a single pulse is 100fs-10ns;

[0014] The multidimensional spectral information is a three-dimensional spectrum, which contains spectral information in three dimensions: laser wavelength, fragment molecular weight, and fragment intensity, and characterizes one or more of the sequence composition, molecular structure, interaction, and dynamic high-level structure of proteins and / or ester biomolecules.

[0015] When protein ions are dissociated, the dissociated fragments include one or more of the free radical fragments a-, b-, c-, x-, y-, and z-. The protein sequence and site modification determination are achieved by matching the fragment ions with the protein theoretical fragments.

[0016] When ester molecules are dissociated, the key information of the dissociated fragments includes one or more of the alkyl chain, double bond, head group, and side chain, so as to accurately identify the molecular structure of the ester isomers.

[0017] The advantages and positive effects of the present invention are:

[0018] The present invention cooperates with the conversion through the joint action of the first on-off control valve, the second on-off control valve, the reflector adjustment system and the adjustment platform system. The laser can be injected from the ultraviolet laser pipeline, or can be injected through the laser high-transmittance window on the five-way joint after being transmitted through the laser transmission support pipeline. At the same time, the vacuum pump can cooperate to evacuate the vacuum vacuum connection pipeline system to balance the vacuum pressure difference with the mass spectrometry system. Therefore, the present invention can couple the functions of vacuum ultraviolet laser and atmospheric pressure ultraviolet laser without stopping the mass spectrometry, and can introduce 10nm-180nm adjustable wavelength extreme ultraviolet laser It is inserted into the mass spectrometry dissociation ion trap to realize ultrafast photodissociation of proteins and other biological molecules by violet lasers of different wavelengths, and the photodissociation fragments are detected by mass spectrometry. Ultimately, while obtaining the action spectrum of extreme ultraviolet light dissociation and the mass spectrum characterization data of dissociation fragments, it is possible to obtain spectral information in three dimensions: wavelength, fragment molecular weight, and fragment intensity. This extreme ultraviolet light dissociation action spectrum-mass spectrometry can characterize the sequence composition, molecular structure, interaction and dynamic high-level structure of biological molecules such as proteins and esters, and can further realize a multidimensional spectroscopy technology with broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is a structural schematic diagram of an application example of the present invention,

[0020] Figure 2 for Figure 1 A in the enlarged view,

[0021] Figure 3 is a structural schematic diagram of another application example of the present invention,

[0022] Figure 4 for Figure 3 Enlarged view of point B in .

[0023] Among them, 1 is a mass spectrometry system, 101 is a vacuum sealing plate, 102 is a first on-off control valve, 2 is an adjustment platform system, 201 is a translation adjustment mechanism, 2011 is a Y-axis moving frame, 2012 is an X-axis moving frame, 202 is a lifting electric cylinder, 203 is a lifting plate, 2031 is a laser system mounting plate, 204 is a rotation drive device, 205 is a horizontal inclination sensor, 206 is a mass spectrometry system support plate, 3 is a laser system, 301 is a laser transmission support pipeline, 302 is a transmission reflector, 4 is a vacuum connecting pipeline system, 401 is a connecting bellows, 402 is a three-way connector A, 403 is a three-way connector B, 404 is a vacuum gauge, 405 is an observation window, 406 is a laser high-transmittance window, 407 is a second on-off control valve, 408 is an ultraviolet laser pipeline, 409 is a five-way connector, 5 is a reflector adjustment system, 501 is a moving device, 502 is a rotating device, 503 is a supporting bellows, 504 is a connecting rod, 505 is an adjusting reflector, 6 is a vacuum pump, and 7 is a sealing window. DETAILED DESCRIPTION

[0024] The present invention will be further described below in conjunction with the accompanying drawings.

[0025] like Figures 1 to 4 As shown, the present invention includes a mass spectrometer system 1, an adjustment platform system 2, a laser system 3, a vacuum connection pipeline system 4 and a reflector adjustment system 5, wherein Figure 1 As shown, the mass spectrometer system 1 and the laser system 3 are both arranged at the upper end of the adjustment platform system 2, and a mass spectrometer dissociation ion trap is arranged inside the mass spectrometer system 1, which is a well-known technology in the art. In the present invention, a through hole coaxial with the center of the mass spectrometer dissociation ion trap annular electrode is arranged on the vacuum sealing plate 101 of the mass spectrometer system 1, and a first on-off control valve 102 is arranged on the through hole, as shown in FIG. Figure 2 As shown, the vacuum connection pipeline system 4 includes a five-way joint 409, and the first interface of the five-way joint 409 is connected to the first on-off control valve 102 through a joint assembly, the lower end of the joint assembly is connected to the vacuum pump 6 through a pipeline, the second interface of the five-way joint 409 is connected to the ultraviolet laser pipeline 408 through the second on-off control valve 407, the ultraviolet laser pipeline 408 is connected to a laser generator, the third interface of the five-way joint 409 is provided with an observation window 405, the fourth interface of the five-way joint 409 is provided with a laser high-transmittance window, and as shown Figures 2-3 As shown, the laser emitted by the laser system 3 is reflected and transmitted by a transmission reflector 302 on a laser transmission support pipeline 301, and then is shot into the five-way joint 409 through the laser high-transmittance window. The reflector adjustment system 5 includes an adjustment reflector 505 and a reflector adjustment mechanism, wherein the adjustment reflector 505 is arranged inside the five-way joint 409, the reflector adjustment mechanism is installed on the fifth interface of the five-way joint 409, and the adjustment reflector 505 adjusts its posture through the reflector adjustment mechanism. Each on-off control valve is a well-known technology in the art and is a commercially available product. In this embodiment, each on-off control valve adopts a plug-in valve. The present invention can also select other suitable pipeline on-off control valves according to actual needs.

[0026] like Figure 2As shown, in this embodiment, the joint assembly includes a connecting bellows 401, a three-way joint A402, a three-way joint B403 and a vacuum gauge 404, wherein the first interface of the three-way joint A402 is connected to the first on-off control valve 102 through the connecting bellows 401, the second interface of the three-way joint A402 is connected to the first interface of the five-way joint 409, the third interface at the lower end of the three-way joint A402 is connected to the first interface of the three-way joint B403, the second interface of the three-way joint B403 is connected to the vacuum gauge 404, the third interface of the three-way joint B403 is connected to the vacuum pump 6 through a pipeline, the vacuum pump 6 is used for vacuuming, and the vacuum gauge 404 is used to detect the vacuum degree inside the pipeline. This is a commercially available product.

[0027] like Figure 2 As shown, in this embodiment, the reflector adjustment mechanism includes a moving device 501, a rotating device 502, a supporting bellows 503 and a connecting rod 504, wherein the rotating device 502 is driven to move by the moving device 501, the lower end of the supporting bellows 503 is mounted on the output shaft of the rotating device 502 and is fixedly connected to the housing of the rotating device 502, the upper end of the supporting bellows 503 is fixedly connected to the fifth interface of the five-way joint 409, and the supporting bellows 503 can be adaptively extended and retracted as the adjusting reflector 505 moves and adjusts, the lower end of the connecting rod 504 is inserted into the supporting bellows 503 and is fixedly connected to the output shaft of the rotating device 502, so that the connecting rod 504 can be driven to rotate by the rotating device 502, and the upper end of the connecting rod 504 is fixedly connected to the adjusting reflector 505.

[0028] In this embodiment, the moving device 501 can be a rodless cylinder, the rotating device 502 is arranged on the moving slide of the rodless cylinder, the rotating device 502 can be a micro servo motor, and the lower end of the supporting bellows 503 is installed on the servo motor housing. The rodless cylinder and the micro servo motor are both well-known technologies in the art and are commercially available products.

[0029] like Figure 1 As shown, in this embodiment, the adjustment platform system 2 includes a translation adjustment mechanism 201, a lifting electric cylinder 202, a lifting plate 203, a rotation drive device 204 and a mass spectrometry system support plate 206, wherein each lifting electric cylinder 202 is arranged on the translation adjustment mechanism 201, each corner end of the lifting plate 203 is respectively hinged to the upper end of the corresponding lifting electric cylinder 202, the rotation drive device 204 is arranged on the lifting plate 203, the mass spectrometry system support plate 206 is arranged on the rotation drive device 204, and the mass spectrometry system support plate 206 is driven to rotate by the rotation drive device 204, the mass spectrometry system 1 is arranged on the mass spectrometry system support plate 206, and the laser system 3 is arranged on one side of the lifting plate 203.

[0030] like Figure 1As shown, in this embodiment, the translation adjustment mechanism 201 includes a Y-direction moving frame 2011, a Y-direction driving device, an X-direction moving frame 2012 and an X-direction driving device. The Y-direction moving frame 2011 is driven by the Y-direction driving device to move along the Y-direction. The X-direction moving frame 2012 and the X-direction driving device are both arranged on the Y-direction moving frame 2011, and the lower side of the X-direction moving frame 2012 is slidably matched with a slide rail arranged on the Y-direction moving frame 2011 through a slider. The X-axis drive device drives the lifting plate 203 to move in the X-axis direction. Four lifting electric cylinders 202 are arranged on the X-axis moving frame 2012, and the power shaft ends of each lifting electric cylinder 202 are respectively hinged with the corresponding ends of the lifting plate 203, so that the four lifting electric cylinders 202 can drive the lifting plate 203 to move up and down when they are synchronously extended and retracted, and can drive the lifting plate 203 to adjust the tilt angle when the four lifting electric cylinders 202 are not synchronously extended and retracted. The lifting plate 203 is used to detect the horizontal tilt angle sensor 205 of its tilt angle. In this embodiment, the Y-axis drive device and the X-axis drive device can both adopt a motor and a lead screw nut structure, wherein the lead screw is driven to rotate by the motor, and the nut is sleeved on the lead screw and fixedly connected to the Y-axis moving frame 2011 or the X-axis moving frame 2012, which is a well-known technology in the art.

[0031] like Figure 1 As shown, in this embodiment, the rotation driving device 204 can adopt a reduction servo motor, and the lower side of the mass spectrometry system support plate 206 is installed on the power shaft of the reduction servo motor, which is a well-known technology in the art.

[0032] like Figure 1 As shown, in this embodiment, a laser system mounting plate 2031 is provided on one side of the lifting plate 203, and the laser system 3 and the laser transmission support pipeline 301 are both provided on the laser system mounting plate 2031, and the moving device 501 of the reflector adjustment mechanism can be fixed on a bracket ( Figure 1 The lower end of the bracket is also fixed on the laser system mounting plate 2031. Figure 3 As shown, in this embodiment, the upper and lower ends of the laser transmission support pipeline 301 are both provided with transmission reflectors 302, and the light emitted by the laser system 3 is reflected by each transmission reflector 302 and then emitted into the five-way joint 409 through the laser high-transmittance window.

[0033] The working principle of the present invention is:

[0034] like Figures 1-2As shown, in an application example of the present invention, the second on-off control valve 407 is closed, and then the vacuum pump 6 is turned on to evacuate the vacuum connecting pipeline system 4. After the vacuum degree of this part reaches the vacuum degree required by the ultraviolet laser pipeline 408 and the mass spectrometry system 1, the second on-off control valve 407 is opened, and the vacuum ultraviolet laser generated by the laser generator is injected into the five-way joint 409 via the ultraviolet laser pipeline 408. At this time, the reflector 505 is adjusted not to rise, and then the operator controls to open the first on-off control valve 102 on the mass spectrometry system 1. At this time, the vacuum degree of the mass spectrometry system 1 is balanced with the vacuum degree of the vacuum connecting pipeline system 4, and then the soft on-off valve 102 of the mass spectrometry system 1 is turned on. The control system controls the vacuum ultraviolet laser to enter the mass spectrometer, wherein the adjustment platform system 2 can fine-tune the inclination angle of the lifting plate 203 through each lifting electric cylinder 202 according to the test needs, and fine-tune the horizontal angle of the mass spectrometer system 1 through the rotary drive device 204 so that the vacuum ultraviolet laser can accurately act on the precursor ion group, further dissociate and ionize the precursor ions, and can also complete the automatic scanning of the multi-degree-of-freedom combination within the specified interval according to the laser dissociation and ionization signal intensity and find the best position for laser dissociation and ionization. After the best position is confirmed, the control system controls and adjusts the locking of various parts of the adjustment platform system 2 to ensure the reliability of the instrument during operation. This application example is used for vacuum ultraviolet photodissociation, and the pressure difference between the ultrahigh vacuum in the mass spectrometer system 1 and the vacuum of the ultraviolet laser pipeline 408 is balanced by evacuating the vacuum connection pipeline system 4.

[0035] like Figures 1-2As shown, another application example of the present invention is an ultraviolet laser propagating under atmospheric pressure. The laser system 3 can be a laser light source such as 193nm, 248nm, 266nm or 355nm. Many different types of ultraviolet or other wavelength lasers can also be used, or multiple beams of lasers with different wavelengths can be introduced into the mass spectrometer through a time pulse delay device, which is not limited to lasers in the ultraviolet band. When the application example is working, the first on-off control valve 102 and the second on-off control valve 407 are both closed, and the adjusting reflector 505 is driven to rise and adjust the angle by the reflector adjustment mechanism, and then the vacuum pump 6 is turned on to evacuate the vacuum connection pipeline system 4. After reaching the set vacuum degree, the first on-off control valve 102 on the mass spectrometry system 1 is slowly opened, and the ultraviolet laser emitted by the laser system 3 is reflected and transmitted through each transmission reflector 302 passing through the laser transmission support pipeline 301 under atmospheric pressure, and then is shot into the adjusting reflector 505 through the laser high-transmittance window at the fourth interface of the five-way joint 409, and then enters the mass spectrometry system 1 after being reflected by the adjusting reflector 505 to dissociate biological molecules such as proteins. In addition, the application example can also add another second laser system to the ultraviolet laser pipeline 408 according to the experimental needs, and introduce another laser beam with a different wavelength emitted by the second laser system into the mass spectrometry system 1 through the time pulse delay setting, and the wavelength of this laser beam is not limited to the ultraviolet band. This application example is used for UV photodissociation, which enables coupling of UV lasers of different wavelengths under atmospheric pressure without stopping the mass spectrometer to dissociate biological molecules such as proteins, greatly improving the efficiency of the experiment.

[0036] like Figures 3-4 As shown, in another application example of the present invention, the outer port of the first on-off control valve 102 can be directly provided with a sealing window 7, and the ultraviolet laser (emitted by the laser system 3) under atmospheric pressure is directly injected through the sealing window 7. The sealing window 7 is a well-known technology in the art. In this application example, the sealing window 7 is installed on the first on-off control valve 102 by screw compression to form a laser vacuum sealing entrance. When this application example is working, the first on-off control valve 102 is slowly opened, and the software system controls the laser emitted by the atmospheric pressure ultraviolet laser system to be directly injected from the sealing window 7 into the mass spectrometer to dissociate biological molecules such as proteins. At the same time, the system can also introduce another laser beam with a different wavelength into the mass spectrometer system 1 through a time pulse delay device, and the wavelength of this laser beam is not limited to the ultraviolet band.

[0037] The present invention can realize a multi-dimensional spectroscopy technology, specifically: introducing a biological sample into the mass spectrometry system 1 through the sample inlet of the mass spectrometry system 1, selecting a specific wavelength extreme ultraviolet laser in the 10nm~180nm band to irradiate the biological molecule ions such as proteins in the mass spectrometry ion trap, and detecting the products of extreme ultraviolet photodissociation and ionization by mass spectrometry; scanning different laser wavelengths in the 10nm~180nm band, and recording the products of photodissociation and ionization at different wavelengths by mass spectrometry, to obtain multi-dimensional spectroscopy information covering both the extreme ultraviolet photodissociation spectrum information of biological molecules and the mass spectrum information of fragment ions, and the 10nm~180nm adjustable wavelength extreme ultraviolet laser can realize single pulse or multiple pulse photodissociation of biological molecules such as proteins, wherein the duration of a single pulse can be 100fs~10ns.

[0038] The multidimensional spectral information is a three-dimensional spectrum, which contains spectral information in three dimensions: laser wavelength, fragment molecular weight, and fragment intensity, and can characterize one or more of the sequence composition, molecular structure, interaction, and dynamic high-level structure of biological molecules such as proteins and / or esters.

[0039] The present invention can efficiently dissociate protein ions. The dissociated fragments are a-, b-, c-, x-, y-, z- and other types of fragments, including one or more free radical fragments. Protein sequence and site modification determination can be achieved by matching the fragment ions with protein theoretical fragments.

[0040] The present invention can efficiently dissociate ester molecules such as phospholipids, and the dissociated fragments contain one or more of the key information such as alkyl chains, double bonds, head groups, and side chains, and can accurately identify the molecular structures of ester isomers.

Claims

1. An instrument for extreme ultraviolet photodissociation spectroscopy-mass spectrometry integrated spectroscopy, comprising a mass spectrometry system, wherein a mass spectrometry dissociation ion trap is provided inside the mass spectrometry system, characterized in that: The invention comprises an adjustment platform system (2), a laser system (3), a vacuum connection pipeline system (4) and a reflector adjustment system (5), wherein the mass spectrometer system (1) and the laser system (3) are both arranged at the upper end of the adjustment platform system (2), a through hole coaxial with the center of the mass spectrometer dissociation ion trap annular electrode is arranged on the vacuum sealing plate (101) of the mass spectrometer system (1), and a first on-off control valve (102) is arranged on the through hole, the vacuum connection pipeline system (4) comprises a five-way joint (409), and a first interface of the five-way joint (409) is connected to the first on-off control valve (102) through a joint assembly, the lower end of the joint assembly is connected to the vacuum pump (6) through a pipeline, the second interface of the five-way joint (409) is connected to the ultraviolet laser pipeline (408) through a second on-off control valve (407), and the five-way joint (409) is connected to the ultraviolet laser pipeline (408) through a second on-off control valve (407). The third interface of the head (409) is provided with an observation window (405), the fourth interface of the five-way joint (409) is provided with a laser high-transmittance window, and the laser emitted by the laser system (3) is reflected and transmitted by a transmission reflector (302) on a laser transmission support pipeline (301) and then injected into the five-way joint (409) through the laser high-transmittance window. The reflector adjustment system (5) includes an adjustment reflector (505) and a reflector adjustment mechanism. The adjustment reflector (505) is arranged inside the five-way joint (409), and the laser injected into the five-way joint (409) is reflected by the adjustment reflector (505) and injected into the mass spectrometry system (1). The reflector adjustment mechanism is installed on the fifth interface of the five-way joint (409), and the adjustment reflector (505) adjusts its posture through the reflector adjustment mechanism.

2. The instrument for extreme ultraviolet photodissociation spectroscopy-mass spectrometry integrated spectroscopy according to claim 1, characterized in that: The joint assembly comprises a connecting bellows (401), a three-way joint A (402), a three-way joint B (403) and a vacuum gauge (404), wherein a first interface of the three-way joint A (402) is connected to a first on-off control valve (102) via the connecting bellows (401), a second interface of the three-way joint A (402) is connected to a first interface of the five-way joint (409), a third interface at the lower end of the three-way joint A (402) is connected to a first interface of the three-way joint B (403), a second interface of the three-way joint B (403) is connected to the vacuum gauge (404), and a third interface of the three-way joint B (403) is connected to the vacuum pump (6) via a pipeline.

3. The instrument for EUV photodissociation spectroscopy-mass spectrometry integrated spectroscopy according to claim 1, characterized in that: The reflector adjustment mechanism comprises a moving device (501), a rotating device (502), a supporting bellows (503) and a connecting rod (504), wherein the rotating device (502) is driven to move by the moving device (501), the lower end of the supporting bellows (503) is sleeved on the output shaft of the rotating device (502) and is fixedly connected to the housing of the rotating device (502), the upper end of the supporting bellows (503) is fixedly connected to the fifth interface of the five-way joint (409), the lower end of the connecting rod (504) extends into the supporting bellows (503) and is fixedly connected to the output shaft of the rotating device (502), and the upper end of the connecting rod (504) is fixedly connected to the adjusting reflector (505).

4. The instrument for EUV photodissociation spectroscopy-mass spectrometry integrated spectroscopy according to claim 1, characterized in that: The adjustment platform system (2) comprises a translation adjustment mechanism (201), a lifting electric cylinder (202), a lifting plate (203), a rotation drive device (204) and a mass spectrometry system support plate (206), wherein each lifting electric cylinder (202) is arranged on the translation adjustment mechanism (201), each corner end of the lifting plate (203) is respectively hinged to the upper end of the corresponding lifting electric cylinder (202), the rotation drive device (204) is arranged on the lifting plate (203), the mass spectrometry system support plate (206) is arranged on the rotation drive device (204), and the mass spectrometry system support plate (206) is driven to rotate by the rotation drive device (204), the mass spectrometry system (1) is arranged on the mass spectrometry system support plate (206), and the laser system (3) is arranged on one side of the lifting plate (203).

5. A method of the instrument for extreme ultraviolet photodissociation spectroscopy-mass spectrometry integrated spectroscopy according to claim 2, characterized in that: The first on-off control valve (102) and the second on-off control valve (407) are closed and the adjustment mirror (505) is not raised. The vacuum pump (6) is first turned on to evacuate the vacuum connection pipeline system (4). When the vacuum degree inside the vacuum connection pipeline system (4) reaches the required level, the second on-off control valve (407) is opened, and the vacuum ultraviolet laser is injected into the five-way joint (409) along the ultraviolet laser pipeline (408). Then, the first on-off control valve (102) on the mass spectrometry system (1) is opened, and the lifting electric cylinder (202) in the platform system (2) is adjusted to fine-tune the inclination angle of the lifting plate (203). The rotary drive device (204) is fine-tuned to adjust the horizontal angle of the mass spectrometry system (1) so that the vacuum ultraviolet laser is injected into the mass spectrometry system (1) and accurately acts on the biological sample to be dissociated.

6. A method of the EUV photodissociation spectroscopy mass spectrometry integrated spectroscopy instrument according to claim 2, characterized in that: The first on-off control valve (102) and the second on-off control valve (407) are closed, and the adjusting reflector (505) is driven to rise and adjust the angle by the reflector adjustment mechanism, and then the vacuum pump (6) is turned on to evacuate the vacuum connection pipeline system (4). When the vacuum degree inside the vacuum connection pipeline system (4) reaches the required level, the first on-off control valve (102) on the mass spectrometry system (1) is opened, and the ultraviolet laser emitted by the laser system (3) passes through the transmission line on the laser transmission support pipeline (301). After being reflected and transmitted by the input reflector (302), the laser is irradiated onto the adjustment reflector (505) through the laser high-transmittance window at the fourth interface of the five-way joint (409), and then enters the mass spectrometry system (1) after being reflected by the adjustment reflector (505). The lifting electric cylinder (202) in the adjustment platform system (2) fine-tunes the tilt angle of the lifting plate (203), and the rotary drive device (204) fine-tunes the horizontal angle of the mass spectrometry system (1) so that the vacuum ultraviolet laser enters the mass spectrometry system (1) and accurately acts on the biological sample to be dissociated.

7. The method of the instrument for EUV-DPDS-MS integrated spectroscopy according to claim 6, characterized in that: The ultraviolet laser pipeline (408) is connected to the second laser system, and through the time pulse delay setting, another laser beam with a different wavelength emitted by the second laser system and the laser beam emitted by the laser system (3) are respectively and alternately introduced into the mass spectrometry system (1), and the adjusting reflector (505) cooperates to adjust the angle so that the laser can accurately act on the biological sample to be dissociated.

8. An application of the instrument for extreme ultraviolet photodissociation spectroscopy-mass spectrometry integrated spectroscopy according to claim 1, characterized in that: The biological sample is introduced into the mass spectrometry system (1) through the sample inlet of the mass spectrometry system (1), and an extreme ultraviolet laser with a specific wavelength is selected in the 10nm to 180nm band to irradiate the biological sample in the mass spectrometry dissociation ion trap through the vacuum connection pipeline system (4), and the products of extreme ultraviolet photodissociation and ionization are detected by mass spectrometry; different laser wavelengths are scanned in the 10nm to 180nm band, and the products of photodissociation and ionization at different wavelengths are recorded by mass spectrometry to obtain multidimensional spectral information covering both extreme ultraviolet photodissociation spectrum information of biological molecules and mass spectrum information of fragment ions, and the 10nm to 180nm adjustable wavelength extreme ultraviolet laser realizes single pulse or multiple pulse photodissociation of protein and / or ester biological molecules in the biological sample, wherein the duration of a single pulse is 100fs to 10ns; The multidimensional spectral information is a three-dimensional spectrum, which contains spectral information in three dimensions: laser wavelength, fragment molecular weight, and fragment intensity, and characterizes one or more of the sequence composition, molecular structure, interaction, and dynamic high-level structure of proteins and / or ester biomolecules.

9. The use of the instrument for extreme ultraviolet photodissociation spectroscopy-mass spectrometry integrated spectroscopy according to claim 8, characterized in that: When protein ions are dissociated, the dissociated fragments include one or more of the free radical fragments a-, b-, c-, x-, y-, and z-. The protein sequence and site modification determination are achieved by matching the fragment ions with the protein theoretical fragments.

10. The use of the instrument for extreme ultraviolet photodissociation spectroscopy-mass spectrometry integrated spectroscopy according to claim 8, characterized in that: When ester molecules are dissociated, the key information of the dissociated fragments includes one or more of the alkyl chain, double bond, head group, and side chain, so as to accurately identify the molecular structure of the ester isomers.

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