An instrument and method for extreme ultraviolet photodissociation spectroscopy mass spectrometry integrated spectroscopy

Abstract

The application relates to an instrument and method of extreme ultraviolet photodissociation spectroscopy mass spectrum integration spectrum, wherein a through hole is arranged on a mass spectrum system, a first on-off control valve is arranged on the through hole, a vacuum connection pipeline system comprises a five-way joint, and each interface of the five-way joint is connected with a joint assembly, a second on-off control valve, an observation window, a laser high-transparency window and a mirror system; the lower end of the joint assembly is connected with a vacuum pump through a pipeline; the second on-off control valve is connected with an ultraviolet laser pipeline; laser is transmitted through a laser transmission pipeline and is shot into the laser high-transparency window; and the mirror in the mirror system is adjustably arranged in the five-way joint. The application can couple the functions of vacuum ultraviolet laser and atmospheric pressure ultraviolet laser under the condition that the mass spectrum does not stop, can introduce adjustable wavelength extreme ultraviolet laser into a mass spectrum dissociation ion trap to realize the ultrafast photodissociation of biological molecules such as proteins, and further realizes a multi-dimensional spectrum technology.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of spectral and mass spectrometry, in particular to an instrument and method for integrating spectrum of extreme ultraviolet photodissociation spectroscopy and mass spectrometry. BACKGROUND

[0002] With the vigorous development of mass spectrometry, mass spectrometry has become a key analytical tool for analyzing proteins and metabolites, and tandem mass spectrometry often includes one or more ion dissociation stages for analyzing samples, which generates product ions by dissociating ions generated by samples, and measures the intensity and mass-to-charge ratio of product ions to clarify or quantify highly specific target or non-target analytes. Current methods for obtaining product ions 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 emitted by an ultraviolet laser to irradiate precursor ion groups to fragment them and generate product ions. This technology can promote more extensive and deep fragmentation of proteins, providing high sequence detection range, and is very suitable for high-throughput proteomics. However, 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 a spectrum with both sample spectral information and mass spectral information, which severely limits the development of ultraviolet photodissociation (UVPD) technology. SUMMARY

[0004] The purpose of the present application is to provide an instrument and method for integrating spectrum of extreme ultraviolet photodissociation spectroscopy and mass spectrometry, which can couple the functions of vacuum ultraviolet laser and atmospheric pressure ultraviolet laser under the condition of uninterrupted mass spectrometry, and can introduce 10nm-180nm tunable wavelength extreme ultraviolet laser into the mass spectrometry dissociation ion trap, realizing ultrafast photodissociation of different wavelength ultraviolet laser on proteins and other biological molecules.

[0005] The purpose of the present application is achieved by the following technical solutions:

[0006] An instrument of extreme ultraviolet photodissociation spectroscopy mass spectrum integration spectrum, comprising a mass spectrum system, a mass spectrum dissociation ion trap is arranged in the mass spectrum system, further comprising an adjusting platform system, a laser system, a vacuum connection pipeline system and a mirror adjusting system, wherein the mass spectrum system and the laser system are arranged on the upper end of the adjusting platform system, a through hole coaxial with the center of the annular electrode of the mass spectrum dissociation ion trap is arranged on the vacuum sealing plate of the mass spectrum system, a first on-off control valve is arranged on the through hole, the vacuum connection pipeline system comprises a five-way joint, the first interface of the five-way joint is connected with the first on-off control valve through a joint assembly, the lower end of the joint assembly is connected with a vacuum pump through a pipeline, the second interface of the five-way joint is connected with an ultraviolet laser pipeline through a 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 transmitted into the five-way joint through a transmission mirror on a laser transmission support pipeline and then enters the five-way joint through the laser high-transmittance window, the mirror adjusting system comprises an adjusting mirror and a mirror adjusting mechanism, the adjusting mirror is arranged in the five-way joint, and the laser entering the five-way joint is reflected into the mass spectrum system through the adjusting mirror, and the mirror adjusting mechanism is installed on the fifth interface of the five-way joint, and the adjusting mirror adjusts the posture through the mirror adjusting mechanism.

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

[0008] The mirror adjusting mechanism comprises a moving device, a rotating device, a support bellows and a connecting rod, wherein the rotating device is driven to move through the moving device, the lower end of the support bellows is sleeved on the output shaft of the rotating device and is fixedly connected with the rotating device shell, the upper end of the support bellows is fixedly connected with the fifth interface of the five-way joint, the lower end of the connecting rod extends into the support bellows and is fixedly connected with the output shaft of the rotating device, and the upper end of the connecting rod is fixedly connected with the adjusting mirror.

[0009] The adjusting platform system comprises a translation adjusting mechanism, lifting electric cylinders, a lifting plate, a rotating drive device and a mass spectrum system support plate, wherein each lifting electric cylinder is arranged on the translation adjusting mechanism, each corner end of the lifting plate is hingedly connected with the upper end of the corresponding lifting electric cylinder, the rotating drive device is arranged on the lifting plate, the mass spectrum system support plate is arranged on the rotating drive device and is driven to rotate through the rotating drive device, the mass spectrum system is arranged on the mass spectrum system support plate, and the laser system is arranged on one side of the lifting plate.

[0010] A method for using the instrument of the extreme ultraviolet photodissociation spectroscopy mass spectrum integration spectrum, specifically: the first on-off control valve and the second on-off control valve are closed, and the adjusting mirror is not raised, the vacuum pump is first started to vacuumize the vacuum connection pipeline system, when the internal vacuum degree of the vacuum connection pipeline system reaches the requirement, the second on-off control valve is opened, the vacuum ultraviolet laser is shot into the five-way joint along the ultraviolet laser pipeline, then the first on-off control valve on the mass spectrum system is opened, and the lifting cylinder in the adjusting platform system is finely adjusted to tilt the angle of the lifting plate, the horizontal angle of the mass spectrum system is finely adjusted by the rotary drive device, so that the vacuum ultraviolet laser is shot into the mass spectrum system and accurately acts on the biological sample to be dissociated.

[0011] A method for using the instrument of the extreme ultraviolet photodissociation spectroscopy mass spectrum integration spectrum, specifically: the first on-off control valve and the second on-off control valve are closed, and the adjusting mirror is not raised, the vacuum pump is first started to vacuumize the vacuum connection pipeline system, when the internal vacuum degree of the vacuum connection pipeline system reaches the requirement, the second on-off control valve is opened, the vacuum ultraviolet laser is shot into the five-way joint along the ultraviolet laser pipeline, then the first on-off control valve on the mass spectrum system is opened, and the lifting cylinder in the adjusting platform system is finely adjusted to tilt the angle of the lifting plate, the horizontal angle of the mass spectrum system is finely adjusted by the rotary drive device, so that the vacuum ultraviolet laser is shot into the mass spectrum system and accurately acts on the biological sample to be dissociated.

[0012] The ultraviolet laser pipeline is connected with the second laser system, and the time pulse delay setting is set to make another laser beam of different wavelength shot by the second laser system and the laser beam shot by the laser system be introduced into the mass spectrum system alternately, and the adjusting mirror is adjusted to make the laser accurately act on the biological sample to be dissociated.

[0013] The application of an instrument integrating mass spectrometry and spectroscopy according to the extreme ultraviolet photodissociation spectrum, specifically: introducing a biological sample into a mass spectrometry system through a sample inlet of the mass spectrometry system, selecting a specific wavelength extreme ultraviolet laser in a 10nm-180nm wave band to irradiate the biological sample in a mass spectrometry dissociation ion trap through the vacuum connection pipeline system, and detecting the products of extreme ultraviolet photodissociation and ionization through mass spectrometry; scanning different laser wavelengths in a 10nm-180nm wave band, and recording the products of photodissociation and ionization under different wavelengths through mass spectrometry to obtain multi-dimensional spectroscopy information covering the extreme ultraviolet photodissociation spectrum information and fragment ion mass spectrometry information of biological molecules, and realizing single pulse or multiple pulse photodissociation of proteins and / or ester biological molecules in a biological sample through a 10nm-180nm adjustable wavelength extreme ultraviolet laser, wherein the single pulse duration is 100fs-10ns.

[0014] The multi-dimensional spectroscopy information is a three-dimensional spectrum, which contains spectroscopy information of three dimensions of laser wavelength, fragment molecular weight and fragment intensity, and characterizes one or more than two of sequence composition, molecular structure, interaction and dynamic higher structure of proteins and / or ester biological molecules.

[0015] When protein ions are dissociated, the dissociation fragments include one or more than two of radical fragments in a-, b-, c-, x-, y- and z-, and the protein sequence and site modification are determined by matching the fragment ions with the theoretical fragments of the protein.

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

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

[0018] The application can couple the functions of vacuum ultraviolet laser and atmospheric pressure ultraviolet laser under the condition that the mass spectrum does not stop, and can introduce 10nm-180nm tunable wavelength extreme ultraviolet laser into the ion dissociation well of the mass spectrum, realize the ultrafast photodissociation of biological molecules such as proteins by different wavelength ultraviolet laser, and detect the photodissociation fragments by the mass spectrum, finally realize that the spectral information of three dimensions of wavelength, fragment molecular weight and fragment intensity is obtained at the same time that the action spectrum of extreme ultraviolet photodissociation and the dissociation fragment mass spectrum characterization data are obtained, the extreme ultraviolet photodissociation action spectrum-mass spectrum can characterize the sequence composition, molecular structure, interaction and dynamic higher structure of biological molecules such as proteins and esters, and can further realize a multi-dimensional spectral technology, and has wide application prospect. BRIEF DESCRIPTION OF DRAWINGS

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

[0020] Figure 2 It is an enlarged view of A in Figure 1

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

[0022] Figure 4 It is an enlarged view of B in Figure 3

[0023] Wherein, 1 is a mass spectrum system, 101 is a vacuum sealing plate, 102 is a first on-off control valve, 2 is an adjusting platform system, 201 is a translation adjusting mechanism, 2011 is a Y-direction moving frame, 2012 is an X-direction moving frame, 202 is a lifting electric cylinder, 203 is a lifting plate, 2031 is a laser system mounting plate, 204 is a rotary driving device, 205 is a horizontal inclination sensor, 206 is a mass spectrum system support plate, 3 is a laser system, 301 is a laser transmission support pipeline, 302 is a transmission mirror, 4 is a vacuum connection pipeline system, 401 is a connecting bellows, 402 is a three-way joint A, 403 is a three-way joint 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 joint, 5 is a mirror adjusting system, 501 is a moving device, 502 is a rotating device, 503 is a support bellows, 504 is a connecting rod, 505 is an adjusting mirror, 6 is a vacuum pump, and 7 is a sealing window piece.​​ Detailed Implementation

[0024] The invention will now be described in further detail with reference to the accompanying drawings.

[0025] like Figures 1 to 4 As shown, the present invention includes a mass spectrometry system 1, an adjustment platform system 2, a laser system 3, a vacuum connection pipeline system 4, and a reflector adjustment system 5, wherein, as... Figure 1 As shown, both the mass spectrometry system 1 and the laser system 3 are located on the upper end of the adjustment platform system 2, and the mass spectrometry system 1 is equipped with a mass spectrometry dissociation ion trap, which is a well-known technology in the art. In this invention, a through hole coaxial with the center of the annular electrode of the mass spectrometry dissociation ion trap is provided on the vacuum sealing plate 101 of the mass spectrometry system 1, and a first on / off control valve 102 is provided on the through hole. Figure 2 As shown, the vacuum connection pipeline system 4 includes a five-way connector 409. The first port of the five-way connector 409 is connected to the first on / off control valve 102 via a connector assembly. The lower end of the connector assembly is connected to the vacuum pump 6 via a pipeline. The second port of the five-way connector 409 is connected to the ultraviolet laser pipeline 408 via a second on / off control valve 407. The ultraviolet laser pipeline 408 is connected to a laser generator. The third port of the five-way connector 409 is provided with an observation window 405, and the fourth port of the five-way connector 409 is provided with a high-transmittance laser window. Figures 2 to 3 As shown, the laser emitted by the laser system 3 is reflected and transmitted through a transmission reflector 302 on a laser transmission support pipe 301, and then enters the five-way connector 409 through the high-transmission laser window. The reflector adjustment system 5 includes an adjustment reflector 505 and a reflector adjustment mechanism. The adjustment reflector 505 is located inside the five-way connector 409, and the reflector adjustment mechanism is installed on the fifth interface of the five-way connector 409. The adjustment reflector 505 is adjusted in attitude by the reflector adjustment mechanism. The various on / off control valves are known in the art and are commercially available products. In this embodiment, each on / off control valve is a plug-in valve. Other suitable pipeline on / off control valves can also be selected according to actual needs.

[0026] like Figure 2As shown in the embodiment, 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 the first interface of the three-way joint A 402 is connected with the first on-off control valve 102 through the connecting bellows 401, the second interface of the three-way joint A 402 is connected with the first interface of the five-way joint 409, the third interface of the lower end of the three-way joint A 402 is connected with the first interface of the three-way joint B 403, the second interface of the three-way joint B 403 is connected with the vacuum gauge 404, the third interface of the three-way joint B 403 is connected with the vacuum pump 6 through a pipeline, the vacuum pump 6 is used for vacuumizing, and the vacuum gauge 404 is used for detecting the vacuum degree inside the pipeline, which is a commercially available product.

[0027] As shown in the embodiment, Figure 2 As shown in the embodiment, the mirror adjusting 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 with the housing of the rotating device 502, the upper end of the supporting bellows 503 is fixedly connected with the fifth interface of the five-way joint 409, the supporting bellows 503 can be adaptively telescoped with the movement of the adjusting mirror 505, the lower end of the connecting rod 504 is inserted into the supporting bellows 503 and is fixedly connected with 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 with the adjusting mirror 505.

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

[0029] As shown in the embodiment, Figure 1 As shown in the embodiment, the adjusting platform system 2 comprises a translation adjusting mechanism 201, lifting cylinders 202, a lifting plate 203, a rotating driving device 204, and a mass spectrum system support plate 206, wherein each lifting cylinder 202 is arranged on the translation adjusting mechanism 201, each corner end of the lifting plate 203 is hingedly connected with the upper end of the corresponding lifting cylinder 202, the rotating driving device 204 is arranged on the lifting plate 203, the mass spectrum system support plate 206 is arranged on the rotating driving device 204 and is driven to rotate by the rotating driving device 204, the mass spectrum system 1 is arranged on the mass spectrum system support plate 206, and the laser system 3 is arranged on one side of the lifting plate 203.

[0030] As shown in the embodiment, Figure 1As shown, in this embodiment, the translation adjustment mechanism 201 includes a Y-axis moving frame 2011, a Y-axis driving device, an X-axis moving frame 2012, and an X-axis driving device. The Y-axis moving frame 2011 is driven to move along the Y-axis by the Y-axis driving device. The X-axis moving frame 2012 and the X-axis driving device are both mounted on the Y-axis moving frame 2011. The lower side of the X-axis moving frame 2012 slides with a slide rail mounted on the Y-axis moving frame 2011 via a slider. Driven by the X-axis drive device, the device moves along the X-axis. Four lifting cylinders 202 are mounted on the X-axis moving frame 2012, and the power shaft end of each lifting cylinder 202 is hinged to the corresponding end of the lifting plate 203. Thus, when the four lifting cylinders 202 extend and retract synchronously, they can drive the lifting plate 203 to rise and fall; when the four lifting cylinders 202 extend and retract asynchronously, they can drive the lifting plate 203 to adjust its tilt angle. The lifting plate 203 is used by a horizontal tilt sensor 205 to detect its tilt angle. In this embodiment, both the Y-axis drive device and the X-axis drive device can adopt a motor and lead screw / nut structure, where the lead screw is driven to rotate by the motor, and the lead screw nut is fitted onto the lead screw and fixedly connected to the Y-axis moving frame 2011 or the X-axis moving frame 2012. This is a well-known technology in the art.

[0031] like Figure 1 As shown, in this embodiment, the rotary drive device 204 can be a geared servo motor, and the mass spectrometry system support plate 206 is mounted on the power shaft of the geared servo motor on its lower side. This 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 pipe 301 are both provided on the laser system mounting plate 2031. The moving device 501 of the reflector adjustment mechanism can be fixed to a bracket. Figure 1 (Not shown in the image), the lower end of the bracket is also fixed to the laser system mounting plate 2031. Additionally, as shown... Figure 3 As shown, in this embodiment, both the upper and lower ends of the laser transmission support pipe 301 are provided with transmission reflectors 302. The light emitted by the laser system 3 is reflected by each transmission reflector 302 and then enters the five-way connector 409 through the high-transmission laser window.

[0033] The working principle of this invention is as follows:

[0034] like Figures 1 to 2As shown, in one application example of the present application, the second on-off control valve 407 is closed, and then the vacuum pump 6 is started to pump the vacuum connection pipeline system 4 to vacuum, and after the vacuum degree of this part reaches the required vacuum degree of the ultraviolet laser pipeline 408 and the mass spectrometry system 1, the second on-off control valve 407 is opened, the vacuum ultraviolet laser generated by the laser generator is emitted into the five-way joint 409 through the ultraviolet laser pipeline 408, at this time the adjusting mirror 505 is not raised, 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 connection pipeline system 4, then the software control system of the mass spectrometry system 1 controls the vacuum ultraviolet laser to enter the mass spectrometry, wherein the adjusting platform system 2 can adjust the tilt angle of the lifting plate 203 through the micro-adjustment of each lifting cylinder 202 according to the test requirements, and the horizontal angle of the mass spectrometry system 1 is micro-adjusted through the rotary driving device 204 to make the vacuum ultraviolet laser accurately act on the precursor ion group, and the precursor ion is further dissociated and ionized, and the automatic scanning of the multi-degree-of-freedom combination in the specified interval can also be completed according to the laser dissociation and ionization signal intensity to find the best position of the laser dissociation and ionization, after the best position is confirmed, the control system controls to adjust the locking of each part of the adjusting platform system 2 to ensure the reliability of the instrument during operation. The application example is used for vacuum ultraviolet photodissociation, and the pressure difference between the ultrahigh vacuum in the mass spectrometry system 1 and the vacuum of the ultraviolet laser pipeline 408 is balanced by pumping the vacuum connection pipeline system 4 to vacuum.

[0035] As Figures 1 to 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 source such as 193nm, 248nm, 266nm or 355nm, or many different types of ultraviolet or other wavelength lasers can be used, or multiple lasers of different wavelengths can be introduced into the mass spectrum through a time pulse delay device, not limited to lasers in the ultraviolet band. In this application example, both the first on / off control valve 102 and the second on / off control valve 407 are closed. The adjusting mirror 505 is raised and its angle adjusted via the mirror adjustment mechanism. Then, the vacuum pump 6 is turned on to evacuate the vacuum connection pipeline system 4. Once the set vacuum level is reached, the first on / off control valve 102 on the mass spectrometry system 1 is slowly opened. The ultraviolet laser emitted by the laser system 3, under atmospheric pressure, is reflected by the various transmission mirrors 302 along the laser transmission support pipeline 301 and then enters the adjusting mirror 505 through the high-transmission window at the fourth interface of the five-way connector 409. After being reflected by the adjusting mirror 505, it enters the mass spectrometry system 1 to dissociate proteins and other biomolecules. Alternatively, this application example can also add a second laser system to the ultraviolet laser pipeline 408 as needed. By setting a time pulse delay, another laser beam of a different wavelength emitted by the second laser system can be introduced into the mass spectrometry system 1. This laser wavelength is not limited to the ultraviolet band. This application example uses ultraviolet photodissociation to couple ultraviolet lasers of different wavelengths under atmospheric pressure to dissociate biomolecules such as proteins without shutting down the mass spectrometer, greatly improving experimental efficiency.

[0036] like Figures 3 to 4 As shown, in another application example of the present invention, a sealing window 7 can be directly installed at the outer port of the first on / off control valve 102, allowing atmospheric pressure ultraviolet laser (emitted by laser system 3) to directly enter through the sealing window 7. The sealing window 7 is a known technology in the art. In this application example, the sealing window 7 is tightened onto the first on / off control valve 102 with screws to form a laser vacuum sealed inlet. During operation, 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 directly enter the mass spectrometer through the sealing window 7 to dissociate proteins and other biomolecules. Simultaneously, the system can also introduce another laser beam of a different wavelength into the mass spectrometer system 1 via a time pulse delay device; the wavelength of this laser beam is not limited to the ultraviolet band.

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

[0038] The multi-dimensional spectroscopy information is a three-dimensional spectrum, which contains spectroscopy information of three dimensions of laser wavelength, fragment molecular weight and fragment intensity, and can characterize one or two or more of sequence composition, molecular structure, interaction and dynamic higher-order structure of biological molecules such as proteins and / or esters.

[0039] The application can efficiently dissociate protein ions, and the dissociation fragments are one or two or more of a-, b-, c-, x-, y-, z- and other types of fragments containing free radical fragments, and the protein sequence and site modification can be determined by matching the fragment ions with the theoretical fragments of the protein.

[0040] The application can efficiently dissociate ester molecules such as phospholipids, and the dissociation fragments contain one or two or more of key information such as alkyl chain, double bond, head group and branched chain, and the ester isomer molecular structure can be accurately identified.

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 arranged inside the mass spectrometry system, characterized in that: The application relates to a mass spectrometry system, which comprises an adjusting platform system (2), a laser system (3), a vacuum connecting pipeline system (4) and a mirror adjusting system (5), wherein the mass spectrometry system (1) and the laser system (3) are arranged on the upper end of the adjusting platform system (2), a through hole coaxial with the center of a mass spectrometry dissociation ion trap ring electrode is arranged on a vacuum sealing plate (101) of the mass spectrometry system (1), a first on-off control valve (102) is arranged on the through hole, the vacuum connecting pipeline system (4) comprises a five-way joint (409), a first interface of the five-way joint (409) is connected with the first on-off control valve (102) through a joint assembly, the lower end of the joint assembly is connected with a vacuum pump (6) through a pipeline, a second interface of the five-way joint (409) is connected with an ultraviolet laser pipeline (408) through a second on-off control valve (407), a third interface of the five-way joint (409) is provided with an observation window (405), a fourth interface of the five-way joint (409) is provided with a laser high-transparency window, laser emitted by the laser system (3) is reflected by a transmission mirror (302) on a laser transmission support pipeline (301) and then enters the five-way joint (409) through the laser high-transparency window, the mirror adjusting system (5) comprises an adjusting mirror (505) and a mirror adjusting mechanism, the adjusting mirror (505) is arranged in the five-way joint (409), and the laser entering the five-way joint (409) is reflected by the adjusting mirror (505) and then enters the mass spectrometry system (1), and the mirror adjusting mechanism is installed on a fifth interface of the five-way joint (409), and the adjusting mirror (505) adjusts the posture through the mirror adjusting mechanism.

2. The instrument of claim 1, wherein: 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 with the first on-off control valve (102) through the connecting bellows (401), a second interface of the three-way joint A (402) is connected with the first interface of the five-way joint (409), a third interface at the lower end of the three-way joint A (402) is connected with a first interface of the three-way joint B (403), a second interface of the three-way joint B (403) is connected with the vacuum gauge (404), and a third interface of the three-way joint B (403) is connected with the vacuum pump (6) through a pipeline.

3. The instrument of claim 1, wherein: The mirror adjusting 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 through 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 with the shell of the rotating device (502), the upper end of the supporting bellows (503) is fixedly connected with 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 with the output shaft of the rotating device (502), and the upper end of the connecting rod (504) is fixedly connected with the adjusting mirror (505). ​ 4. The instrument of claim 1, wherein: The adjusting platform system (2) comprises a translation adjusting mechanism (201), lifting cylinders (202), a lifting plate (203), a rotary driving device (204) and a mass spectrometry system support plate (206), wherein each lifting cylinder (202) is arranged on the translation adjusting mechanism (201), each corner end of the lifting plate (203) is hingedly connected to the upper end of a corresponding lifting cylinder (202), the rotary driving device (204) is arranged on the lifting plate (203), the mass spectrometry system support plate (206) is arranged on the rotary driving device (204) and is driven to rotate by the rotary driving device (204), and 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. The method of claim 2, wherein the instrument is an extreme ultraviolet photodissociation spectroscopy mass spectrometry integrated spectroscopy instrument. The first on-off control valve (102) and the second on-off control valve (407) are closed, the adjusting mirror (505) is not raised, the vacuum pump (6) is first started to vacuumize the vacuum connection pipeline system (4), when the vacuum degree in the vacuum connection pipeline system (4) reaches the requirement, the second on-off control valve (407) is opened, the vacuum ultraviolet laser is shot 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 cylinder (202) in the adjusting platform system (2) finely adjusts the tilt angle of the lifting plate (203) and the rotary driving device (204) finely adjusts the horizontal angle of the mass spectrometry system (1) so that the vacuum ultraviolet laser is shot into the mass spectrometry system (1) and accurately acts on the biological sample to be dissociated.

6. A method of an extreme ultraviolet photodissociation spectroscopy mass spectrometry integrated spectrometry instrument according to claim 2, wherein: The first on-off control valve (102) and the second on-off control valve (407) are closed, the adjusting mirror (505) is raised and adjusted in angle by the mirror adjusting mechanism, then the vacuum pump (6) is started to vacuumize the vacuum connection pipeline system (4), when the vacuum degree in the vacuum connection pipeline system (4) reaches the requirement, the first on-off control valve (102) on the mass spectrometry system (1) is opened, the ultraviolet laser emitted by the laser system (3) is reflected by the transmission mirror (302) on the laser transmission support pipeline (301) and then is shot into the adjusting mirror (505) through the laser high-transmittance window at the fourth interface of the five-way joint (409), and then is reflected by the adjusting mirror (505) and enters the mass spectrometry system (1), the lifting cylinder (202) in the adjusting platform system (2) finely adjusts the tilt angle of the lifting plate (203), and the rotary driving device (204) finely adjusts the horizontal angle of the mass spectrometry system (1) so that the vacuum ultraviolet laser is shot into the mass spectrometry system (1) and accurately acts on the biological sample to be dissociated.

7. The method of claim 6, wherein the instrument is an extreme ultraviolet photodissociation spectroscopy mass spectrometry integrated spectrometer. The ultraviolet laser pipeline (408) is connected to the second laser system, and another laser beam of different wavelength emitted by the second laser system is alternately introduced into the mass spectrometry system (1) with the laser beam emitted by the laser system (3) through time pulse delay setting, and the adjusting mirror (505) is adjusted in angle to make the laser accurately act on the biological sample to be dissociated.

8. Use of an 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) from the sample inlet of the mass spectrometry system (1), a specific wavelength extreme ultraviolet laser in the 10nm-180nm wave band is selected to irradiate the biological sample in the mass spectrometry dissociation ion trap through the vacuum connection pipeline system (4), and the products of the extreme ultraviolet photodissociation and ionization are detected by mass spectrometry; different laser wavelengths in the 10nm-180nm wave band are scanned, and the photodissociation and ionization products at different wavelengths are recorded by mass spectrometry to obtain multi-dimensional spectroscopy information covering the extreme ultraviolet photodissociation spectrum information and the fragment ion mass spectrum information of the biological molecules, and the 10nm-180nm adjustable wavelength extreme ultraviolet laser realizes single pulse or multiple pulse photodissociation of the protein and / or ester biological molecules in the biological sample, wherein the single pulse duration is 100fs-10ns. The multi-dimensional spectroscopy information is a three-dimensional spectrum, which contains spectroscopy information of three dimensions of laser wavelength, fragment molecular weight and fragment intensity, and characterizes one or more than two of the sequence composition, molecular structure, interaction and dynamic higher-order structure of the protein and / or ester biological molecules.

9. The use of an instrument for extreme ultraviolet photodissociation spectroscopy mass spectrometry hybrid spectroscopy according to claim 8, characterized in that: When the protein ion is dissociated, the dissociation fragments include one or more than two of a-, b-, c-, x-, y-, z- containing radical fragments, and the protein sequence and site modification are determined by matching the fragment ions with the theoretical fragments of the protein.

10. The use of an instrument for extreme ultraviolet photodissociation spectroscopy mass spectrometry hybrid spectroscopy according to claim 8, characterized in that: When the ester molecule is dissociated, the key information of the dissociation fragments includes one or more than two of alkyl chain, double bond, head group and branch, so as to accurately identify the molecular structure of the ester isomer.

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