A more complete appreciation of the present invention and many attendant advantages thereof will be readily obtained as the same becomes better understood by reference to the following detailed description when considered in connection with the accompanying drawings, wherein:
There is a need for high throughput LC-MS analysis using high-resolution MS instruments. In the present invention, multiple LC columns are interfaced to a multi-channel MS. According to one embodiment of the present invention, a QqTOF(or QTOF) MS instrument provides the multi-channel MS. QTOF in this application offers high mass accuracy (2-3 ppm), reasonable size, and cost efficiency due to a high degree of sharing major subsystems between all channels.
QqTOF-MS offers the highest mass accuracy comparable to that of FTICR-MS (2-3 ppm), high throughput (due to pulsing ions into oaTOF with high frequency up to 10 kHz), reasonable size (desktop QqTOF systems are commercially available) and cost efficiency. In one embodiment of the present invention, subsystems of QqTOF-MS are shared between channels to reduce cost and complexity even more.
In another embodiment of the present invention, a multi-channel ESI-QqTOF mass spectrometer having many common channel parts/subsystems (i.e., shared between the channels) provides a MS system with reasonable size without compromising on instrument performance including mass resolution, mass accuracy, and dynamic range (including channel cross-talk). Sample throughput in these units will be proportional to the number of the channels. In one embodiment of the present invention, each channel operation can be completely independent of other channels, thus making the MS system operation the same as if a single-channel QqTOF-MS was used.
Referring now to the drawings, wherein like reference numerals designate identical, or corresponding parts throughout the several views, and more particularly to
While the configurations of the multi-channel QqTOF-MS are shown in
Briefly illustration, in operation of the MS systems 10, 50 of the present invention, ions are provided from separate ion source 12, 52 such as for exampled separate ESI ion sources or any other sources like MALDI, atmospheric (AP) MALDI, AP chemical ionization (APCI), or AP photoionization (APPI). The ions enter the MS systems 10, 50 by inlets 14, 54 such as for example separate heated capillaries of 0.5-1 mm diameter. The ions enter into the first vacuum region 16, 56 (e.g., a 1 Torr vacuum region) in which a first set of quadrupoles 18, 58 operating for example at the 1 Torr pressure focus and direct ions further into a second vacuum region 20, 60 (e.g, a 0.1 Torr vacuum region). While not shown in
After collisional cooling in the quadrupoles in the second vacuum region 20, 60, a second set of quadrupoles 22, 62 operating for example at the 1 Torr pressure in the second vacuum regions 20, 60 can focus and direct ions further into a third vacuum region 24, 64 (e.g, a 1 mTorr vacuum region). The ions upon being transferred into the third vacuum region 22, 62 are acted on by a third set of quadrupoles 26, 66. Ions in the third vacuum region 24, 64 are further acted on by conductance limits/diaphragms (shown in
An ion isolation quadrupole and collisional chamber 40 for ion fragmentation permitting MS/MS spectra analysis can be included in one embodiment of the present invention as shown in
The way to apply both trapping RF and broadband (AC) excitation waveforms to the isolation quadrupole Q1 is shown in
After leaving the quadrupole section, the ion beam is focused and slightly steered by ion optical lenses 42 (like the Einzel lens in
The pulser device 32, 72 can utilize high repetition and fast front edge high voltage pulses to push ions into the orthogonal reflectron (typically 5-10 kHz, typically up to 1 kV amplitude or more). The pulser device 32, 72 can be shared between all channels. Instead of periodic push-out pulses, the pulses in the accelerator can be pseudo-randomly spread in time (i.e., the so called Hadamard oa-reTOF design).
The design of the vacuum system in one embodiment of the present invention serves to minimize cross-talk between the channels and permits a more dynamic range to be achieved. The vacuum system design in one embodiment of the present invention is shown in
The first vacuum region 16 in the multi-channel system 10 can be pumped using for example a roots blower/rotary vane pump system 110 (e.g., Model 2033+RSV 301 B by Adixen-Alcatel, France,) offering for example a pumping speed up to 300 m3/hr at 1 Torr pressure). To minimize cross-talk in the high pressure areas where a substantial gas and ion cross flow may take place, the first and second vacuum regions 16 and 20 can be separated into subsections pumped by the same pumps but through separate vacuum hoses (e.g., connecting between ports 1, 2, 3, and 4 as shown in
The gas load on the third vacuum region 24 and main vacuum region 28 (e.g., the 1 mTorr and 10−7 Torr regions) can be minimized by using quadrupoles instead of multipole ion guides with a smaller rod diameter to reduce the conductance limit between the high vacuum regions and, thus reducing the pumping load on the turbo pumps. The region of the pulser device and the reflectron (i.e. the 10−7 Torr region) may require an additional turbo pump to maintain the required vacuum.
Second-order focusing upon an initial spatial distribution of ions across the ion beam in the pulser device 30 permits, in one embodiment of the present invention, high (e.g., usually, more than 10,000) resolving power in the Qq-reTOF system. There are several approaches in the present invention to achieve second order space focusing. In one approach, for the oe-reTOF configuration, a single-stage extraction is used in conjunction with a two-stage ion mirror (reflectron). Alternatively, a two-stage extraction and a single-stage reflectron can be used. Both configurations permit adjusting two experimental parameters (i.e., the electric field strengths in two reflectron stages in the former approach and the extraction field and the reflecting field in the latter case) to zero the first and second expansion term in the dependence of the ion time-of-flight over the initial ion position in the pulser region.
In one embodiment of the present invention, the dependence of the total time-of-flight of an ion over the initial ion position in the pulser region is expanded into a series having the expansion terms of first, second, third, etc. orders over the deviation of the initial ion position from the ion beam axis. Then, by tuning the experimental parameters (usually by tuning the voltages as described above) the first and second expansion terms are zeroed, thus, achieving minimal dependence of the ion flight time on the initial ion position in the beam. Such techniques available to the present invention are referred to as a first (if the first term is zeroed only) or second order space focusing (if both the first and second terms are zeroed).
In principle, in the multi-channel schemes shown in the present invention, it is possible to obtain the second order space focusing simultaneously in all channels if all channels are substantially identical. This is because the geometry of time-of-flight section is the same for all channels. In practice, one can expect slight deviations from the ideal geometry due to manufacturing tolerances, accuracy of assembly and alignment. To tolerate small imperfections in the channel fabrication and accuracy of assembly, one embodiment of the present invention permits the control of at least one parameter separately for each channel.
Since both tunable parameters in the single-stage extraction/double-stage reflectron approach are related to the reflectron which is shared between all channels, one embodiment of the present invention uses a double-stage extraction/single-stage reflectron scheme shown in
One method for adjustment is based on experimentally observed relative insensitivity of the space focusing conditions on the tune parameters (−GiVa and Vref). This is due to presence of other factors limiting the mass resolution in QqTOF-MS, among which is the so-called “turn-around” effect due to a small transverse velocity of ions in the beam entering the pulser region. The “turn-around-time” actually determines the final system resolution (after the second-order space focusing is achieved). For this reason, typically no channel-specific adjustment of the grid voltage −GiVa is required.
In one embodiment of the present invention, ions can be detected using microchannel plate (MCP) 180 (as shown in
If the time interval between the arrivals of two or more ions is shorter than the dead time, the peak shape obtained by summation of all ion hit events may be distorted and so mass accuracy may be compromised. The ADC systems (digitizers) do not have inherent dead time. Modern 8-bit ADC's with real-time on-board averaging have digitizing rate of several gigahertz (for example, a FASTFLIGHT Digital Signal Averager from Signal Recovery, Oak Ridge, Tenn.).
While described in detail above, the present invention can be considered more generally to include the following embodiments not limited to the specific features described above.
In one embodiment of the present invention, the parallel mass spectroscopy sampling of the present invention is directed to protemics applications which can benefit from the present techniques providing high mass accuracy and high throughput.
In one embodiment of the present invention, the parallel mass spectroscopy sampling of the present invention is provided for in a novel mass spectrometry system in which components of the system are shared in common between the multi-channels.
In one embodiment of the present invention, the parallel mass spectroscopy sampling of the present invention is provided for in a novel system including an orthogonal acceleration time-of-flight mass analyzer having at least two channels for ion introduction into the mass analyzer, a field-free section configured to mass-separate ions during flight time, and an ion detector for detection of ions transiting the field-free region. The two channels are configured to receive respective groups of ions in separate ones of the channels for mass analysis of the respective groups of ions. At least a part of the field-free section is shared between the two channels.
In one embodiment of the present invention, the parallel mass spectroscopy sampling of the present invention is provided for in a novel multi-channel oa-reTOF mass spectrometry system in which the reflectron is shared, as it typically represents one of the largest components in the oa-reTOF mass spectrometer. However, other components of the oa-reTOF mass spectrometer can be shared as well.
In one embodiment of the present invention, ions can be produced in at least two ion sources and thereafter can be directed to different channels of the multi-channel orthogonal extraction (oe) reflectron TOF (reTOF) mass spectrometer in which the two channels share for example a common reflectron in their operation.
In another embodiment of the present invention, the two channels can share an orthogonal accelerator of ions.
In another embodiment of the present invention, a common orthogonal accelerator can extract ions into the common reflectron with a frequency in the range from 10 Hz to 100 kHz.
In another embodiment of the present invention, the mass spectrometer can include a data system which acquires mass spectra corresponding to each of the two channels.
In another embodiment of the present invention, the data system can include at least one microchannel plate (MCP) ion detector which operates in an ion counting or mocke on ion current measurement mode.
In another embodiment of the present invention, the ions can be time-focused on the microchannel plate detector using at least one of a first order space focusing and a second order space focusing.
In another embodiment of the present invention, the microchannel plate detector can have multiple anodes, used for detecting ions directed to different channels.
In another embodiment of the present invention, the two channels can include atmospheric pressure (AP) interfaces and ion guides to deliver ions to the common accelerator and reflectron and the atmospheric pressure interfaces. The ion guides can share common DC and RF power supplies.
In another embodiment of the present invention, the mass spectrometer can be placed into a multi-section vacuum chamber with differentially-pumped sections and there sections can be shared between the two channels.
In another embodiment of the present invention, the mass spectrometer can include at least one of a single stage oe-reTOF mass analyzer, a double stage oe-reTOF mass analyzer, a multipass oe-reTOF mass analyzer, and a Hadamard oe-reTOF mass analyzer.
In another embodiment of the present invention, the mass spectrometer can include at least one of an ion isolation means and an ion fragmentation means.
In another embodiment of the present invention, the ion isolation device can include at least one of a quadrupole RF/DC mass filter, a broad-band excitation mass filter, a stored waveform isolation Fourier transform (SWIFT) isolation, and a notched broad-band excitation.
In another embodiment of the present invention, the ion fragmentation device can include at least one of a collision-induced dissociation, a surface-induced dissociation, an electron capture dissociation, an electron transfer dissociation, and a metastable atom-induced dissociation.
One feature of the present invention is the utilization of the pulser device to re-direct ions from for example a horizontal direction to a vertical direction, as shown in the figures. In one embodiment of the present invention, once the ions are re-directed, the ions are then mass analyzed to identify the constituents of those ions that originated from the respective ion sources. While a reflectron time-of-flight analyzer has been shown for the mass analyzer, the mass analyzer in one embodiment of the present invention could be a linear time-of-flight analyzer. In other embodiments of the present invention, the mass analyzer receiving ions from pulser could be a quadrupole ion trap (QIT), a toroidal ion trap, a linear quadrupole ion trap, an orbitrap, a Fourier transform mass spectrometer (FT-MS), an ion mobility spectrometer (IMS), a high-field asymmetric, waveform ion mobility spectrometer (FAIMS), and their combination(s).
At step 610, the ion sources can be one of an electrospray ionization (ESI) source, an atmospheric pressure chemical ionization (APCI) source, an atmospheric pressure photo-ionization (APPI) source, a matrix-assisted laser desorption/ionization (MALDI) source, and a atmospheric pressure (AP) MALDI source. At 600,
At 620, the ions can be directed into a reflection shared by at least two ion introduction channels in the mass analyzer. At 620, the ions can be directed into an orthogonal accelerator is shared between the at least two channels. At 620,
At 630, a data acquisition system can acquire mass spectra corresponding to the group of ions introduced in the channels. At 630, the data acquisition system can operate an ion counting mode or an ion current measurement mode. At 630, mass spectra can be obtained from a tandem (MS/MS or MSn) mass selection. At 630, a quadrupole RF/DC mass filter, a broad-band excitation mass filter, a stored waveform isolation Fourier transform (SWIFT) isolation mass filter, and/or a notched broad-band excitation mass filter can be used for ion isolation in the mass analyzer. At 630, the ions can be fragments in a collision-induced dissociation unit, a surface-induced dissociation unit, an electron capture dissociation unit, an electron transfer dissociation unit, a fast atom bombardment dissociation unit, and a metastable atom-induced dissociation unit.
Numerous modifications and variations on the present invention are possible in light of the above teachings. It is, therefore, to be understood that within the scope of the accompanying claims, the invention may be practiced otherwise than as specifically described herein.