CN100521064C - Matrix auxiliary laser decomposition-absorption-particle bundle collision decomposition flying time mass spectrograph - Google Patents
Matrix auxiliary laser decomposition-absorption-particle bundle collision decomposition flying time mass spectrograph Download PDFInfo
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- CN100521064C CN100521064C CNB2007100099214A CN200710009921A CN100521064C CN 100521064 C CN100521064 C CN 100521064C CN B2007100099214 A CNB2007100099214 A CN B2007100099214A CN 200710009921 A CN200710009921 A CN 200710009921A CN 100521064 C CN100521064 C CN 100521064C
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Abstract
A base material assisting laser desorption-particle beam collision dissociation time-of-flight mass spectrometer relates to a mass spectrograph and provides a time-of-flight mass spectrometer obviously decreasing noise disturbance and improving mass resolution. The invention is provided with a cavity, a sample introduction probe rod, a preposition and a postposition mesh units, a particle beam generator, a multi-polar rod, a particle beam detector, a microchannel plate electron multiplier, a laser and a focusing lens. The sample introduction probe rod, the preposition mesh unit, the postposition mesh unit, the particle beam generator, the multi-polar rod, the particle beam detector, the microchannel electron multiplier are equipped inside the cavity in turn. The cavity and the sample introduction probe rod are coaxial. The axle wire of the postposition mesh unit is coaxial with the particle beam generator, the multi-polar rod and the particle beam detector. The cavity is provided with a quartz window for laser in, a vacuum opening for extraction and a valve for sample introduction probe rod in and out on the wall. The sample introduction probe rod is provided with a sampling fixing component on the top end. The laser and the focusing lens are equipped outside the cavity.
Description
Technical field
The present invention relates to a kind of mass spectrometer, especially relate to a kind of substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates.
Background technology
At present, technology commonly used comprises gel electrophoresis and mass spectrum etc. in the large biological molecule analysis.Mass spectrum has high-resolution, high precision and highly sensitive characteristics.Wherein, substance assistant laser desorpted ion source (MALDI) and electric spray ion source (ESI) have obtained extensive use in the large biological molecule analysis.For daltonian large biological molecules up to a million, though the MALDI technology can be carried out ionization to it, its signal value very a little less than, be difficult to detect.In the ESI source, large biological molecule exists with the form of multi-charge, and resulting collection of illustrative plates is very complicated, is difficult to resolve.And most mass analyzer as quadrupole rod, sector field and ion trap etc., all has the quality upper limit, is generally several kilodaltons.
Flight time mass spectrum is more special, and theoretically, it does not have the quality upper limit, and has the resolution capability up to 10ppm.Yet, because the large biological molecule of super high molecular weight is slower at the flight time mass spectrum medium velocity, on microchannel plate electron multiplier detector, be difficult to detect, thus the detection sensitivity of greatly reducing.Therefore, flight time mass spectrum is subject to and analyzes 200,000～300, the large biological molecule of 000Da.Fuerstenau S.D. etc. attempted once that flight time mass spectrum was used for molecular weight and reached daltonian macromolecular detections up to a million, though this molecule can demonstrate very weak peak at mass spectrogram, still can't reach the analysis requirement, did not have practical value.Larson etc. study the signal difference of different-energy ion at the mass spectral microchannel plate electron multiplier of line time, and the result shows that energy is ion (5kDa, 1.5 * 10 of 6kV
4M/s) it is ion (5kDa, 3 * 10 of 25kV that signal value has only energy
4M/s) half.The corresponding signal of this explanation microchannel plate electron multiplier is finally determined by the speed of ion.Therefore, be 1,000,000 daltonian big molecule for molecular weight, when its speed is 3 * 10
4During m/s, need apply about 5 * 10
6The accelerating voltage of V just might make it obtain detecting.Consider the restriction of factors such as vacuum degree, instrument, can not realize under the actual conditions up to 5 * 10
6The accelerating voltage of V.Therefore, carrying out qualitatively by determining molecular weight to the super large molecule, method is infeasible.Theoretically, can carry out qualitative accurately to it by the fragment information of super large molecule.At present, Chang Yong inducing molecule dissociation technique comprises mainly that collision auxiliary dissociate (CAD), photodissociation, X ray dissociate, electron excitation and spatial induction dissociate etc.Wherein, the auxiliary dissociation technique of collision is the highest for the dissociation efficiency of large biological molecule, therefore is most widely used.In the collision pond, neutral gaseous molecular is not subjected to the control of ion lens, freely diffuses out the collision pond, and near detector is produced very strong noise jamming.
Chinese patent CN85104052 provides a kind of time-of-flight mass spectrometer, and it comprises the analyzer that is provided with many ring electrodes along same axle, and wherein, to used a voltage that is inversely proportional to distance by measured ion, this voltage produces electric field force.
Publication number provides a kind of series connection linear ion hydrazine and time-of-flight mass spectrometer for the application for a patent for invention of CN1926657, and wherein, ion trap has the straight line central shaft with mass spectrometric flight track quadrature.Ion trap comprises: one group of electrode (401,403,402,404), and at least one described electrode has the opening that is used for to the mass spectrometer emitting ions; One group of dc voltage power supply (+V ,-V, V1, V2), be used to provide discrete DC level, and the high-speed electronic switch of some (409), be used to make the DC power supply to be connected/to disconnect with at least two described electrodes; Neutral gas is filled ion trap; And digitial controller, be provided for the handoff procedure of ion trap, operation ion, cooling, and comprise state from ion trap to mass spectrometer that launch all ions from.
Summary of the invention
The object of the present invention is to provide a kind ofly can obviously reduce noise jamming, the substance assistant laser desorpted-particle bundle collision of the resolution that improves the quality dissociates time-of-flight mass spectrometer.
Purpose of the present invention is achieved through the following technical solutions: substance assistant laser desorpted-particle bundle collision time-of-flight mass spectrometer that dissociates is provided with cavity, sample introduction feeler lever, preposition aperture plate group, rearmounted aperture plate group, particle beam generator, multipole bar, particle beams detector, microchannel plate electron multiplier, laser and condenser lens.
Sample introduction feeler lever, preposition aperture plate group, rearmounted aperture plate group, particle beam generator, multipole bar, particle beams detector and microchannel plate electron multiplier are located at inside cavity successively, and the axis of the axis of cavity and sample introduction feeler lever is same axis; The aperture plate parallel interval of preposition aperture plate group is also vertical with the cavity axis; The aperture plate parallel interval of rearmounted aperture plate group is also vertical with the cavity axis, and the axis of the axis of rearmounted aperture plate group and particle beam generator, multipole bar and particle beams detector is same axis; The chamber body lumen wall is provided with quartz window, the vacuum pumping port that confession vacuumizes usefulness and the valve that passes in and out for the sample introduction feeler lever that confession laser is injected; Sample introduction feeler lever top is provided with the sample fixed part; The condenser lens of laser and corresponding laser is located at the cavity outside.
Cavity is preferably T-shaped cylindric stainless steel cavity, and the wall thickness of cavity is preferably 1.5mm, and quartz window and vacuum pump are taken out the bottom that mouth preferably is located at cavity.The sample introduction feeler lever is preferably the stainless steel cylinder, and the diameter of sample introduction feeler lever is preferably 5～20mm, is located at the sample fixed part on sample introduction feeler lever top and establishes at least 1 sample holddown groove.The vacuum degree of cavity is preferably 1～2 * 10
-6Torr.Laser is preferably selected wavelength 157～1100nm, average power for use〉1W, peak power〉100kW, pulsewidth 100fs～10ns, pulse energy 100 μ J～500mJ, pulse frequency 10Hz～10kHz, beam diameter 1～10mm.The technical parameter of condenser lens can be matched according to actual needs.Particle beam generator can be ion gun, electronic generator or X ray transmitter, is preferably ion gun, and energy is 10～10000eV, and ion gun can adopt inert gas ion sources such as He, Ne, N
2, O
2Isoreactivity gas ion source or Cs
+, Li
+, Na
+, K
+, Rb
+In the alkali metal ion source, particle beams energy 5～1000eV, electric current are 1nA～20 μ A, and particle beams diameter is 0.5～20mm, and operating distance is 5～300mm.Preposition aperture plate group is made of 3 aperture plates, and spacing is 1～5mm, and voltage is 1～30kV, and is vertical with the cavity axis.Rearmounted aperture plate group is made of 2 aperture plates, 2 aperture plates and cavity axis normal, and the spacing of 2 aperture plates is 1～5mm, while ground connection.The shape of aperture plate is the stainless steel substrates of rectangle (30～200) mm * (40～220) mm, and the centre is a rectangular hole (15～50) mm * (40～150) mm.The netting twine of aperture plate adopts molybdenum filament, and the diameter of molybdenum filament is 20～80 μ m.Multipole bar can be quadrupole rod, sextupole bar, ends of the earth bar, 12 utmost point bars or ten sextupole bars, and multipole bar is by single solid steel bar construction, and the diameter of every bar of multipole bar is 2～20mm, and bar length is 30～250mm.Particle beams detector can adopt Faraday cup or outsourcing, and the size of Faraday cup is preferably diameter 5～50mm, high 20～75mm.The major parameter of microchannel plate electron multiplier is preferably slab-thickness 0.5～1mm, distance 10～50 μ m between the passage, the resistance 10 between the dull and stereotyped two sides
8～10
10Ω, dull and stereotyped area＜30cm
2
The course of work of the present invention is as follows: whole mass spectrometer all is in high vacuum environment, adopts substance assistant laser desorpted ion source to be used for the ionization of sample molecule.Whole system is divided into 3 zones: be ionization and introducing district in sample introduction feeler lever and the preposition aperture plate group between last 1 aperture plate, be the field-free flight district between the most preceding 1 aperture plate in last 1 aperture plate and the rearmounted aperture plate group in the preposition aperture plate group, separate the abscission zone for particle bundle collision between the most preceding 1 aperture plate and last 1 aperture plate in the rearmounted aperture plate group.Sample is fixed on the sample introduction feeler lever, stretches into cavity by valve.After the laser beam line focus lens focus that laser penetrates, the quartz window by the ion source cavity is mapped to sample surfaces.Substrate molecule absorbs laser energy and gasifies rapidly, and involved sample molecule also is brought into gas phase.The substrate molecule of being excited simultaneously, produces ionization with proton translocation to sample molecule.Sample ions is introduced into the field-free flight district under the electric field action of preposition aperture plate group, separate according to its mass-to-charge ratio.Separate the abscission zone by particle bundle collision successively through the sample ions of separating.The particle beams that particle beam generator produces is vertically injected cavity after multipole bar focuses on, bump with sample ions.Sample ions is dissociated into less fragment ion, and it is detected to strike the microchannel plate electron multiplier.
Operation principle of the present invention is as follows: introduce the field-free flight district after the sample molecule ionization under electric field action, separate preferably according to mass-to-charge ratio.When its successively when the particle bundle collision dissociation, bump with particle in the particle beams, dissociating produces the feature fragment ion, it is detected to strike the microchannel plate electron multiplier.Can realize macromolecular qualitative by measuring these feature fragment ions.Adopt the particle beams to replace neutral gas to realize dissociating of sample ions.In the particle bundle collision dissociation process, the density of impingment particle is The key factor.Ion beam is a most important class particle beams.Can provide enough ion cloud density to guarantee desirable dissociation effect as the ion gun of ion beam generator.Theoretical Calculation proves, for daltonian super large molecules up to a million, total collision frequency of ion and sample ions is equivalent to even more than the collision frequency of neutral gas and sample ions in the conventional tandem mass spectrum collision pond in the particle bundle collision pond.The ion motion that molecular weight is bigger is slow, and is longer by the time in particle bundle collision zone, therefore can obtain more fragment ions.By these fragment information, can carry out observational measurement to big molecule, thereby overcome the quantitative limitation of conventional sense device molecule.Because the ion of big molecular refraction is more, also causes the increase of signal on the detector.By the particle beams detector of vertically placing with ion gun, can also observe the variation of particle beams intensity.The variation of particle beams detector signal value is only relevant with collision process, and is irrelevant with the size of sample molecule.But adopt the quick response electron multiplier of handle high current to improve detection resolution greatly.Because collision area is 1cm only, comparing its length with the field-free flight district of 1m can ignore, and collision and the process of dissociating occur in after the mass separation, so mass resolution can remain on the higher level, and the parsing to spectrogram simultaneously can not cause extra interference.
The relative prior art of the present invention has following outstanding advantage and effect: formed little fragment ion by quick particle bundle collision after 1) big molecule separates through mass-to-charge ratio, can realize qualitative analysis by measuring fragment information.2) adopt the quick particle beams to replace neutral gas to dissociate, reduced background noise to realize collision.3) can be observed the variation of particle beams intensity by particle beams detector.
Description of drawings
Fig. 1 is the structural representation of the embodiment of the invention.
Embodiment
The invention will be further described below in conjunction with drawings and Examples.
The embodiment of the invention is provided with cavity 1, sample introduction feeler lever 2, preposition aperture plate group 3, rearmounted aperture plate group 4, particle beam generator 5, multipole bar 6, particle beams detector 7, microchannel plate electron multiplier 8, laser 9 and condenser lens 10.Sample introduction feeler lever 2, preposition aperture plate group 3, rearmounted aperture plate group 4, particle beam generator 5, multipole bar 6, particle beams detector 7 and microchannel plate electron multiplier 8 are located at cavity 1 inside successively, and the axis of the axis of cavity 1 and sample introduction feeler lever 2 is same axis; Preposition aperture plate group 3 is made of 3 parallel interval and the aperture plate vertical with cavity 1 axis; Rearmounted aperture plate group 4 is made of 2 parallel interval and the aperture plate vertical with cavity 1 axis, and the axis of the axis of rearmounted aperture plate group 3 and particle beam generator 5, multipole bar 6 and particle beams detector 7 is same axis; Cavity 1 chamber wall is provided with quartz window 11, the vacuum pumping port 12 that confession vacuumizes usefulness and the valve 13 that passes in and out for sample introduction feeler lever 2 that confession laser is injected; Sample introduction feeler lever 2 tops are provided with the sample fixed part; The condenser lens 10 of laser 9 and corresponding laser is located at cavity 1 outside.Cavity 1 is T-shaped cylindric stainless steel cavity, and the wall thickness of cavity 1 is 1.5mm, and quartz window 11 and vacuum pump are taken out the bottom that mouth 12 is located at cavity 1.
Sample introduction feeler lever 2 is the stainless steel cylinder, and the diameter of sample introduction feeler lever 2 is 5～20mm, is located at the sample fixed part on sample introduction feeler lever 2 tops and establishes at least 1 sample holddown groove.The vacuum degree of cavity 1 is 1～2 * 10
-6Torr.Laser 9 is selected wavelength 157～1100nm, average power for use〉1W, peak power〉100kW, pulsewidth 100fs～10ns, pulse energy 100 μ J～500mJ, pulse frequency 10Hz～10kHz, beam diameter 1～10mm.The technical parameter of condenser lens 10 can be matched according to actual needs.
Preposition aperture plate group 3 is made of 3 aperture plates, and spacing is 1～5mm, and voltage is 1～30kV, and is vertical with the cavity axis.Rearmounted aperture plate group 4 is made of 2 aperture plates, 2 aperture plates and cavity axis normal, and the spacing of 2 aperture plates is 1～5mm, while ground connection.The shape of aperture plate is the stainless steel substrates of rectangle (30～200) mm * (40～220) mm, and the centre is a rectangular hole (15～50) mm * (40～150) mm.The netting twine of aperture plate adopts molybdenum filament, and the diameter of molybdenum filament is 20～80 μ m.
Multipole bar 6 can be quadrupole rod, sextupole bar, ends of the earth bar, 12 utmost point bars or ten sextupole bars, and multipole bar is by single solid steel bar construction, and the diameter of every bar of multipole bar is 2～20mm, and bar length is 30～250mm.
Particle beams detector 7 adopts Faraday cup or outsourcing, and Faraday cup is of a size of diameter 5～50mm, high 20～75mm.
The major parameter of microchannel plate electron multiplier 8 is slab-thickness 0.5～1mm, distance 10～50 μ m between the passage, the resistance 10 between the dull and stereotyped two sides
8～10
10Ω, dull and stereotyped area＜30cm
2
Claims (9)
1. substance assistant laser desorpted-particle bundle collision time-of-flight mass spectrometer that dissociates is characterized in that being provided with cavity, sample introduction feeler lever, preposition aperture plate group, rearmounted aperture plate group, particle beam generator, multipole bar, particle beams detector, microchannel plate electron multiplier, laser and condenser lens;
Sample introduction feeler lever, preposition aperture plate group, rearmounted aperture plate group, particle beam generator, multipole bar, particle beams detector and microchannel plate electron multiplier are located at inside cavity successively, and the axis of the axis of cavity and sample introduction feeler lever is same axis; The aperture plate parallel interval of preposition aperture plate group is also vertical with the cavity axis; The aperture plate parallel interval of rearmounted aperture plate group is also vertical with the cavity axis, and the axis of the axis of rearmounted aperture plate group and particle beam generator, multipole bar and particle beams detector is same axis; The chamber body lumen wall is provided with quartz window, the vacuum pumping port that confession vacuumizes usefulness and the valve that passes in and out for the sample introduction feeler lever that confession laser is injected; Sample introduction feeler lever top is provided with the sample fixed part; The condenser lens of laser and corresponding laser is located at the cavity outside;
The particle beams that particle beam generator produces is vertically injected cavity after multipole bar focuses on, and the described particle beams and sample ions bump, and sample ions is dissociated into less fragment ion.
2. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, it is characterized in that cavity is T-shaped cylindric stainless steel cavity, the wall thickness of cavity is 1.5mm, quartz window and vacuum pump are taken out the bottom that mouth is located at cavity; The vacuum degree of cavity is 1～2 * 10
-6Torr.
3. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, it is characterized in that the sample introduction feeler lever is the stainless steel cylinder, the diameter of sample introduction feeler lever is 5～20mm, is located at the sample fixed part on sample introduction feeler lever top and establishes at least 1 sample holddown groove.
4. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, wavelength 157～the 1100nm that it is characterized in that laser, average power〉1W, peak power〉100kW, pulsewidth 100fs～10ns, pulse energy 100 μ J～500mJ, pulse frequency 10Hz～10kHz, beam diameter 1～10mm.
5. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, it is characterized in that particle beam generator is ion gun, electronic generator or X ray transmitter, the energy of ion gun is 10～10000eV, and ion gun adopts He, Ne inert gas ion source, N
2, O
2Active gases ion source or Cs
+, Li
+, Na
+, K
+, Rb
+The alkali metal ion source, particle beams energy 5～1000eV, electric current are 1nA～20 μ A, and particle beams diameter is 0.5～20mm, and operating distance is 5～300mm.
6. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, it is characterized in that preposition aperture plate group is made of three aperture plates, spacing is 1～5mm, voltage is 1～30kV, and is vertical with the cavity axis.Rearmounted aperture plate group is made of two aperture plates, two aperture plates and cavity axis normal, the spacing of these two aperture plates is 1～5mm, while ground connection, the shape of aperture plate is the stainless steel substrates of rectangle (30～200) mm * (40～220) mm, the centre is a rectangular hole (15～50) mm * (40～150) mm, and the netting twine of aperture plate adopts molybdenum filament, and the diameter of molybdenum filament is 20～80 μ m.
7. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, it is characterized in that multipole bar can be quadrupole rod, sextupole bar, ends of the earth bar, 12 utmost point bars or ten sextupole bars, multipole bar is by single solid steel bar construction, the diameter of every bar of multipole bar is 2～20mm, and bar length is 30～250mm.
8. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, it is characterized in that particle beams detector adopts Faraday cup, Faraday cup is of a size of diameter 5～50mm, high 20～75mm.
9. as claimed in claim 1 substance assistant laser desorpted-the particle bundle collision time-of-flight mass spectrometer that dissociates, the parameter that it is characterized in that the microchannel plate electron multiplier is slab-thickness 0.5～1mm, distance 10～50 μ m between the passage, the resistance 10 between the dull and stereotyped two sides
8～10
10Ω, dull and stereotyped area＜30cm
2
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