CN1779457A - Gas-phase molecular ion reactor based on radio-frequency quard-polar rod, its realization and use - Google Patents
Gas-phase molecular ion reactor based on radio-frequency quard-polar rod, its realization and use Download PDFInfo
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- CN1779457A CN1779457A CN 200510100350 CN200510100350A CN1779457A CN 1779457 A CN1779457 A CN 1779457A CN 200510100350 CN200510100350 CN 200510100350 CN 200510100350 A CN200510100350 A CN 200510100350A CN 1779457 A CN1779457 A CN 1779457A
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Abstract
A method for realizing gas phase molecule-ion reaction based on radio frequency quadrupole rod includes imposing electric field on said quadrupole rod to let ion analyzed obtain kinetic energy and to let it generate molecule-ion reaction with background gas molecule to generate subion of ion to be analyzed, using mass analyzer to measure subion for realizing cascade mass spectrum analysis of ion to be analyzed. A device of gas phase molecule-ion reactor based on radio frequency quadrupole rod is also disclosed.
Description
Technical field
The present invention relates to be used for the molion reaction technology in tandem mass spectrometry field, particularly a kind of gas molecule in space ionic reaction apparatus and implementation method and application based on radiofrequency quadrupole.
Background technology
In the mass spectrophotometry field, in order to analyze the composition of lower content in the biased sample, the background ions of high concentration composition need be removed, at this moment need the ion filter technology.Simultaneously, increase along with molecular weight analyte to be analyzed, isomers is very common, the structural information of molecule is more and more showed its importance, as adduction position in the body metabolism product of the structure of protein, amino acid sequence of polypeptide, medicine, medicine, poison pollutant in the adduction position of dna fragmentation, DNA base sequence etc., and the molion reaction can provide the structural information of ion, is one of most important parts of tandem mass spectrometry.
The molion reaction of tandem mass spectrometry method general using in gas phase causes analyte ions (parent ion) division and obtains daughter ion, thereby and distributes by the mass spectrum of measuring daughter ion and to obtain the structural information of analyte ions.In the present existing gas phase molion reactor technology by the reaction energy size can be divided into energetic encounter (～keV) and low energy collision (～40eV), these collisions generally occur in the ion cyclotron resonance device, also can occur in ion trap or in quadrupole rod.The energy presentation mode can be that straight line quickens, the axial field in the quadrupole rod quickens and quadrupole rod in two utmost points or four utmost points excite etc.
These existent method respectively have its relative merits, and different ranges of application is arranged.The straight line accelerated method parent ion accelerate to certain energy again with the background gas molecular collision, but owing in accelerator, there is not the quality selectivity, need before collision, add a quadrupole rod that air pressure is different, select interested parent ion and make it to enter reactor, otherwise all ions of different mass-to-charge ratioes enter reactor together, dissociation reaction takes place simultaneously, the spectrum unscrambling difficulty.Because rf electric field or magnetic field can fetter daughter ion effectively improves sensitivity significantly, general molion reaction all occurs in ion trap and the quadrupole rod.The commodity cascade mass spectrometer is general at present adopts a quadrupole mass spectrometer as the one-level mass spectrum, the ion of selecting a kind of mass-to-charge ratio by and straight line accelerate to certain energy and enter a quadrupole rod collision cell, the dissociate daughter ion that obtains of collision is collected by a quadrupole rod and is delivered to the second order ms analyzer.Like this, the air pressure of quadrupole rod collision cell is all higher than former and later two quadrupole rods, needs differential pumped vacuum systems.
Summary of the invention
The objective of the invention is to overcome the shortcoming of prior art, a kind of gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole is provided; This device can optionally excite the ion of certain mass-to-charge ratio, makes it to obtain to collide required kinetic energy, and ionic dissociation rate controllability is good, highly sensitive.
Another object of the present invention is to provide the implementation method of above-mentioned gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole.
It is above-mentioned based on the gas molecule in space ionic reaction apparatus of radiofrequency quadrupole and the application of implementation method thereof that a further object of the present invention is to provide.
Purpose of the present invention is achieved through the following technical solutions: a kind of gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole, be connected with mass analyzer, comprise radiofrequency quadrupole, it is characterized in that: described radiofrequency quadrupole is arranged in the higher gas reaction cavity of air pressure, every bar of radiofrequency quadrupole is formed by three sections, make radiofrequency quadrupole form three grades, the first order is a mass filter, the second level is the molion reactor, the third level is the ion focus transmitter, is provided with the gap between three grades of radiofrequency quadrupoles.Between per two bars of radiofrequency quadrupole, be provided with the axial field electrode that forms axial electric field; The quantity of described axial field electrode is four, and is arranged evenly axisymmetricly.
Gap between three grades of the described radiofrequency quadrupoles is less than 2mm.
The inscribed circle diameter d that four bars of every shank diameter D of described radiofrequency quadrupole and radio frequency form
1Between ratio be about 1.147.The axial field electrode is mounted to a low-angle vertically, i.e. four inscribed circle diameter d that electrode forms
2Littler at the ion inlet end than the endpiece of ion, d
2With d
1Ratio be about 1.3～1.5.
Described gas reaction cavity is provided with air guide port; Described air guide port can be connected with kapillary, and reacting gas can be introduced by kapillary.
A kind of method of utilizing above-mentioned gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole to realize, it is characterized in that: on the radiofrequency quadrupole under certain air pressure, apply exciting voltage, make the analyte ions in the radiofrequency quadrupole obtain kinetic energy, and react with background gas molecule generation molion, produce the daughter ion of analyte ions, and utilize mass analyzer to measure daughter ion, thereby realize cascade mass spectrometry to analyte ions.
Described background gas can be introduced the gas reaction cavity with analyte ions, also can introduce the gas reaction cavity by kapillary.
Apply radio-frequency voltage between two adjacent bars of described radiofrequency quadrupole, three grades of added radio-frequency voltages of quadrupole rod are identical; And apply exciting voltage between the two relative bars, and just wherein relative one group adds sine voltage, and another group adds cosine wave (CW), and two groups of electric voltage frequencies are identical, and amplitude is identical, 90 ° of phase differential.
Described exciting voltage has two groups, is respectively sinusoidal and acts on the two relative bars with cosine wave (CW).Use sinusoidal and cosine excites simultaneously, under identical machinery and electrical quantity, can produce and double two utmost points and excite the energy that excites with four utmost points.
The dc offset voltage of quadrupole rod has two DC voltage, and one is axial field voltage, and one is the quadrupole rod bias voltage.Quadrupole rod bias voltage, radio-frequency voltage and exciting voltage are added on the quadrupole rod by after transformer coupled.
Described background gas is generally the good dry pure gas of chemical stability, as: other stable or inert gases such as helium, nitrogen, argon gas; Background gas air pressure is for being generally 0.1Pa～10Pa.
Described mass analyzer is vertical introduction-type high-resolution flying time mass spectrum analysis device, also can be the mass analyzer of other kinds.
Described analyte ions is baroque organic compound ion (as medicine), life macromolecule (as protein, polypeptide, gene segment) plasma.
Above-mentioned gas molecule in space ionic reaction apparatus and implementation method thereof based on radiofrequency quadrupole can be used for selective excited ion, makes it to obtain to collide required kinetic energy, promptly has only the ion of certain mass-to-charge ratio can obtain daughter ion, and other ions then intactly pass through; And the parent ion of big quality that can dissociate; All right impurity screening ion or high concentration component ion, promptly described first order mass filter not only can only allow a kind of ion of mass-to-charge ratio pass through, and can be to stop ions a certain or a few kinds of mass-to-charge ratioes to pass through.
Action principle of the present invention is: establish m
i/ q, V
Rf, ω, r
0, V
ExcAnd ω
ExcBe respectively radio-frequency voltage, rf frequency, radiofrequency quadrupole inscribed circle radius, exciting voltage and the stimulating frequency of parent ion mass-to-charge ratio, radiofrequency quadrupole.When exciting electric field frequency to be
The time, parent ion obtains maximum kinetic energy.Because the kinetic energy that obtains in quadrupole rod of ion is corresponding one by one with the frequency of exciting voltage, selects different stimulating frequencies, just can select the parent ion of different mass-to-charge ratioes to make it to obtain kinetic energy and also dissociate at last, thereby this device has the quality selectivity; If selected ion obtains enough big kinetic energy within a short period of time, then get on the wall, can not pass through, at this moment play massenfilter.
The present invention has following advantage and effect with respect to prior art:
(1) compares to that two utmost points excite or four utmost points excite and can make parent ion obtain bigger kinetic energy.
(2) ionic dissociation rate controllability is good, and is highly sensitive.
(3) whole device does not need extra differential vacuum system in same cavity.
(4) have the quality selectivity, can selecting necessarily as required, the ion of mass-to-charge ratio carries out the molion reaction.
(5) can impurity screening ion or high concentration component ion, also can only allow certain ion pass through.
Description of drawings
Fig. 1 is the structural representation of apparatus of the present invention.
The cross-sectional view of Fig. 2 device shown in Figure 1.
Fig. 3 is direct current, radio frequency and exciting voltage coupling synoptic diagram.
Fig. 4 is the amino acid sequence synoptic diagram of polypeptide sample to be analyzed.
Fig. 5 is the typical polypeptide mass spectrogram that is obtained by the present invention.
Fig. 6 is that the major component ion is filtered and the spectrogram that obtains in the mass spectrogram 5 (a).
Embodiment
The present invention is described in further detail below in conjunction with embodiment and accompanying drawing, but embodiments of the present invention are not limited thereto.
Fig. 1 shows structure of the present invention, and as seen from Figure 1, this links to each other with mass analyzer 8 by outlet 7 based on gas molecule in space ionic reaction apparatus of radiofrequency quadrupole, is placed on together in the difference grade vacuum system 9; This device comprises radiofrequency quadrupole, and described radiofrequency quadrupole is arranged in the reaction cavity 2, and the vacuum tightness in the reaction chamber can be entered the mouth by ion and 1 regulate, and reacting gas can be introduced from air guide port 3 by kapillary.Every bar of radiofrequency quadrupole is formed by three sections, makes radiofrequency quadrupole form three grades, and the first order 4 is a mass filter, and the second level 5 is the molion reactor, and the third level 6 is provided with the gap for the ion focus transmitter between three grades of radiofrequency quadrupoles; Fig. 2 is shown sectional view of the present invention, is provided with the axial field pole piece 10 that forms axial electric field between per two bars of radiofrequency quadrupole 11; Described axial field pole piece quantity is four, and is arranged evenly axisymmetricly; The diameter D of the every bar of radiofrequency quadrupole and four inscribed circle diameter d that bar forms
1Between ratio be about 1.147; The inscribed circle diameter d that the axial field electrode forms
2With d
1Ratio be about 1.5.
Between two adjacent bars of described radiofrequency quadrupole, apply radio-frequency voltage, and apply exciting voltage between the two relative bars.Described exciting voltage has two groups, is respectively sinusoidal and acts on the two relative bars with cosine wave (CW), and two groups of electric voltage frequencies are identical, and amplitude is identical, but 90 ° of phase differential.Fig. 3 (a) has represented the coupling scheme of quadrupole rod radio-frequency voltage, dc offset voltage, and three kinds of voltages are coupled by three coil transformers respectively; DC voltage is added in the central point of four utmost point radio-frequency transformer (RFT) secondary coils, and exciting voltage and radio-frequency voltage are coupled by other two groups of coil transformers, obtain four output terminals 4 (a), 4 (b), 4 (c) 4 (d), these four output terminals are added in respectively on four bars, and the relative position of four bars is shown in Fig. 3 (b).
The method of utilizing said apparatus to realize is: apply exciting voltage on the radiofrequency quadrupole under certain air pressure, make the analyte ions in the radiofrequency quadrupole obtain kinetic energy, and react with background gas molecule generation molion, produce the daughter ion of analyte ions, and utilize mass analyzer to measure daughter ion, thereby realize cascade mass spectrometry to analyte ions; Specifically: the ion sudden from ion gun at first enters filtering medium quadrupole rod (mass filter), by applying certain frequency, and can be with the major impurity ion remaval; When analyte ions enters into second quadrupole rod (molion reactor), apply the stimulating frequency of this ion, make it to take place the molion reaction, form fragmention; Fragmention forms an ion beam that phase space is less again after entering the 3rd quadrupole rod (ion focus transmitter), by drawing in this device, obtains mass spectrogram by the mass analyzer analysis at last.
This device and method is applied to the polypeptide sample is carried out analyzing and processing, measures the peptide molecule structure, Fig. 4 has represented amino acid sequence of polypeptide figure to be analyzed, and it is arranged in order twice and be combined into ring-type by 5 seed amino acids and constitutes; O, V, P, F, L represent ornithine (Ornithine), valine (Valine), proline (Proline), phenylalanine (Phenylalanine) and leucine (Leucine) respectively among the figure.After utilizing electron spray with its ionization, recording the doubly charged ion mass-to-charge ratio of this polypeptid belt at (shown in Fig. 5 (a)) in the one-level mass spectrogram is 571 ([M+2H]
2+); According to added radio-frequency voltage V on the radiofrequency quadrupole
Rf(170V
P-p), rf frequency ω (2*1.18MHz), the inscribed circle radius r of radiofrequency quadrupole
0(7mm), promptly can calculate the pairing excitation frequency omega of this ion
ExcBe 59.9kHz; With amplitude V
ExcBe adjusted in about 0.8V
P-pAnd be added on second quadrupole rod, when this ion during, under the effect of exciting voltage, obtain kinetic energy through this quadrupole rod, and with the background gas molecular collision, dissociating at last obtains daughter ion, the quadrupole rod air pressure in the present embodiment is approximately 0.16Pa; Shown in Fig. 5 (b), because the various possible fracture fragmentions of parent ion all are focused quadrupole rod and collect, and record by second order ms, by analyze each daughter ion mass spectra peak and with the amino acid masses table relatively, can obtain the amino acid sequence of each fragment ion peak representative.For example: owing to detect mass-to-charge ratio is 197 daughter ion, can find that it is made up of two amino acid of PV; Owing to detect mass-to-charge ratio is 311 daughter ion, can find that it is made up of three amino acid of PVO; Again because to detect mass-to-charge ratio be 424 daughter ion, can find that it is made up of four amino acid of PVOL; So just can affirm, contain PVOL four seed amino acids in this polypeptide, and have the sequence of PVOL.In like manner can detect it and contain other kinds combination, infer the amino acid sequence that this polypeptide is complete at last.
The device of present embodiment is with embodiment 1, handled polypeptide sample is also identical with embodiment, utilize this device and implementation method mainly to carry out the analysis of foreign ion, because this polypeptide ion is lost interest in, want to analyze its impurity, and polypeptide is the principal ingredient of sample, and is influential to the analysis of impurity, wishes before entering mass analyzer its removal; Concrete operations are: make among Fig. 1 three quadrupole rod 4,5,6 added radio-frequency voltages identical, but quadrupole rod 5,6 does not add exciting voltage, at this moment quadrupole rod 5,6 only plays ion transfer and focuses on; And first order quadrupole rod 4 is as massenfilter, and selecting mass-to-charge ratio is 571 ion excitation frequency, and amplitude is higher, as 5V, makes the ion that is excited because the too big kinetic energy of acquisition is got on the wall of quadrupole rod disappears.Mass spectrogram after parent ion is filtered is compared with Fig. 5 (a) as shown in Figure 6, and other all foreign ions are detected, and does not pass through quadrupole rod and mass-to-charge ratio is 571 ion.Select different stimulating frequencies can filter the ion of one or more mass-to-charge ratioes.
Claims (9)
1, a kind of gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole, link to each other with mass analyzer, comprise radiofrequency quadrupole, it is characterized in that: described radiofrequency quadrupole is arranged in the same gas reaction cavity, and every bar of radiofrequency quadrupole is formed by three sections, makes radiofrequency quadrupole form three grades, the first order is a mass filter, the second level is the molion reactor, and the third level is the ion focus transmitter, is provided with the gap between three grades of radiofrequency quadrupoles.
2, the gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole according to claim 1 is characterized in that: the gap between three grades of the described radiofrequency quadrupoles is less than 2mm.
3, the gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole according to claim 1 is characterized in that: be provided with the axial field electrode that forms axial electric field between per two bars of radiofrequency quadrupole; The quantity of described axial field electrode is four, and is arranged evenly axisymmetricly; The inscribed circle diameter d that the axial field electrode forms
2Inscribed circle diameter d with four bars formation of radio frequency
1Ratio be about 1.3～1.5, near the ion end diameter d that enters the mouth
2Less, near ion outlet one end diameter d
2Bigger.
4, a kind of each described method that realizes based on the gas molecule in space ionic reaction apparatus of radiofrequency quadrupole of claim 1～3 of utilizing, it is characterized in that: on the radiofrequency quadrupole under certain air pressure, apply exciting voltage, make the analyte ions in the radiofrequency quadrupole obtain kinetic energy, and react with background gas molecule generation molion, produce the daughter ion of analyte ions, and utilize mass analyzer to measure daughter ion, thereby realize cascade mass spectrometry to analyte ions.
5, the method that realizes based on the gas molecule in space ionic reaction apparatus of radiofrequency quadrupole according to claim 4, it is characterized in that: three grades of quadrupole rods are in the same reaction cavity, and air pressure is identical.
6, the method that realizes based on the gas molecule in space ionic reaction apparatus of radiofrequency quadrupole according to claim 4 is characterized in that: apply radio-frequency voltage between two adjacent bars of described radiofrequency quadrupole, and apply exciting voltage between the two relative bars.
7, the method that realizes based on the gas molecule in space ionic reaction apparatus of radiofrequency quadrupole according to claim 6, it is characterized in that: described exciting voltage has two groups, is respectively sinusoidal and acts on the two relative bars with cosine wave (CW).
8, the method that realizes based on the gas molecule in space ionic reaction apparatus of radiofrequency quadrupole according to claim 4, it is characterized in that: described analyte ions is baroque organic compound ion, life macromolecule plasma.
9, the application of the method that realizes according to each described gas molecule in space ionic reaction apparatus based on radiofrequency quadrupole of claim 4～8, it is characterized in that: be used for selective excited ion, make it to obtain to collide required kinetic energy, or impurity screening ion or high concentration component ion.
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| CN102856153A (en) * | 2012-10-08 | 2013-01-02 | 复旦大学 | Ion optical off-axis transmission system |
| CN106066359A (en) * | 2015-04-24 | 2016-11-02 | 萨默费尼根有限公司 | For the method using the extensive stability property component analysis of quadrupole mass filter |
| CN106872601A (en) * | 2017-03-01 | 2017-06-20 | 暨南大学 | A kind of electron impact ion source carrier gas ion removes device |
| CN109580759A (en) * | 2017-09-28 | 2019-04-05 | 成都飞机工业（集团）有限责任公司 | A kind of quadrupole mass spectrometer |
| CN112944764A (en) * | 2019-12-11 | 2021-06-11 | 博西华电器（江苏）有限公司 | Gas purification device, purification method by gas purification device and refrigerator |
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| JP2004192844A (en) * | 2002-12-09 | 2004-07-08 | Hitachi High-Technologies Corp | Plasma ion source three-dimensional quadrupole mass spectroscope |
| CN2589978Y (en) * | 2002-12-27 | 2003-12-03 | 华南理工大学 | High-resolution quadrupole mass spectrometer |
| JP2005276451A (en) * | 2004-03-23 | 2005-10-06 | Shimadzu Corp | Mass spectrometry device |
| CN1305101C (en) * | 2004-10-27 | 2007-03-14 | 东南大学 | Micro-chamber four-electrode spectrum tube |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102856153A (en) * | 2012-10-08 | 2013-01-02 | 复旦大学 | Ion optical off-axis transmission system |
| CN106066359A (en) * | 2015-04-24 | 2016-11-02 | 萨默费尼根有限公司 | For the method using the extensive stability property component analysis of quadrupole mass filter |
| CN106066359B (en) * | 2015-04-24 | 2020-10-02 | 萨默费尼根有限公司 | Method for broad stability mass analysis using quadrupole mass filters |
| CN106872601A (en) * | 2017-03-01 | 2017-06-20 | 暨南大学 | A kind of electron impact ion source carrier gas ion removes device |
| CN109580759A (en) * | 2017-09-28 | 2019-04-05 | 成都飞机工业（集团）有限责任公司 | A kind of quadrupole mass spectrometer |
| CN112944764A (en) * | 2019-12-11 | 2021-06-11 | 博西华电器（江苏）有限公司 | Gas purification device, purification method by gas purification device and refrigerator |
| CN114242560A (en) * | 2021-11-02 | 2022-03-25 | 中国原子能科学研究院 | Laser photolysis device and method for removing isobaric elements |
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| CN100454020C (en) | 2009-01-21 |
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