Patent Grant
8115167
This application claims priority to U.S. utility application Ser. No. 11/441,766, filed May 26, 2006 and to U.S. provisional application Ser. No. 60/685,247, filed on May 27, 2005.
The present invention relates generally to instrumentation and methodology for characterization of chemical samples based on ion mobility spectrometry (IMS) and mass spectrometry (MS). Specifically, the invention relates to improved IMS, using the concepts of multi-beam ion pre-selection from a single sample, independent yet coordinated separation of each ion beam by both ion mobility and by single or multiple time-of-flight mass spectrometers (TOFMS) (independently for different beams) each with controllable fragmentation of ion mobility separated ions, and multi-channel data recording in one or a few TOFMS. Specifically, the improvements include simultaneously producing and extracting positive and negative ions from the sample (including the accumulation and post-ionization of neutral species), trapping ions in multiple traps, improved transportation of ions against drift gas flow, a new design of mobility cell/TOFMS interface comprising well collimated supersonic cooled gas flows and an original multi-channel RF-ion guide. Additionally, the technique of multi-beam data recording provides methods for increasing the dynamic range of the measurements and for obtaining additional shape analysis beyond that available from ion mobility alone. These improvements may be used to increase throughput from a sample to downstream instruments and methods. The resulting instruments and methods are useful for qualitative and/or quantitative chemical and biological analysis.
An ion mobility spectrometer typically comprises an ionization source, a drift cell, and an ion detector. Examples of an ion detector include a sampling plate, an electron multiplier, or a mass spectrometer. Ion mobility spectrometry separates ions in terms of their mobility in a drift/buffer gas by measuring the ion equilibrium drift velocity. When gaseous ions in the presence of the drift gas experience a constant electric field, they accelerate until the occurrence of a collision with a neutral atom or molecule within the drift gas. This acceleration and collision sequence is repeated continuously. Over time, this microscopic scenario averages the instantaneous velocities over the macroscopic dimensions of the drift tube resulting in the measurement of a constant ion velocity based upon ion size, charge and drift gas pressure. The ratio of the ion velocity to the magnitude of the electric field is defined as ion mobility. In other words, the ion drift velocity (νd) is proportional to the electric field strength (E), where the ion mobility K=νd/E is a function of ion volume/charge ratio. Thus IMS is a separation technique similar to mass spectrometry. IMS is generally known to have high sensitivity with moderate resolving power. Separation efficiency is compromised when “bands” of ions spread apart as opposed to arriving together at the end of the IM drift tube in a tight, well-defined spatial region.
The resolving power for a uniform or quasi-uniform ion mobility electric field increases as a square root of voltage applied along mobility cell. It would seem that there is not much freedom to increase the resolution. However, the situation may be improved if the ion drift in a gas flow is considered. Ions move against the gas flow only if the field is stronger than a certain value specific for the mobility of the ions. Ions with lower mobility may be stationary or even move in the negative direction (with the gas flow). Therefore, better ion separation can be expected where the time of this separation can be chosen suitable for specific applications and compatible with the time diagram of the ion detector operation. The problem is how to efficiently organize ion mobility separation using gas counter-flow. Most often an ion mobility separation is used with ion sources working under elevated pressure and the source pressure is often used when these ions are introduced into a mobility cell. There may be no gas counter-flow in such an application. On the other hand, drift gas counter-flow is inevitable when IMS is used for analysis of ions created in high vacuum ion sources such as a secondary ion source where secondary ions are created from a surface maintained in high vacuum and must then be moved against a counter-flow of gas into the ion mobility spectrometer. The main problem then is how to overcome the strong counter-flow and preserve ion throughput. It is quite natural to use a time varying electric field to gradually move ions from a pulsed ion formation region against the gas flow and into the IMS. Small ions need a relatively small field to overcome the gas flow without decomposing whereas larger ions can come to the entrance orifice later under the action of a stronger field. At the time of application of the larger field necessary to move the heavier ions, small ions are already inside the mobility cell and are not subjected to the strong field which would otherwise cause their fragmentation. Some separation of ions in addition to the usual mobility separation is achieved in this case, however, it is often rather small, because of the diffusion broadening during the initial ion cloud formation. The gas counter-flow itself is also useful because it prevents neutral species from getting into the mobility cell and degrading its performance by forming non-conductive deposits on the mobility cell electrodes. One of the crucial points for the present invention is the organization of the weaker counter-flow for the low pressure ion sources and purposeful creation of the counter-flow for high pressure ion sources for their interfacing with ion mobility cells.
The combination of an ion mobility spectrometer (IMS) with a mass spectrometer (MS) is well known in the art. In 1961, Barnes et al. were among the first to combine these two separation methods. Such instruments allow for separation and analysis of ions according to both their mobility and mass, which is often referred to as two-dimensional separation or two-dimensional analysis. Young et al. realized that an orthogonal time-of-flight mass spectrometer (oTOFMS) is the most preferred mass spectrometer type to be used in such combination because of its ability to detect simultaneously and very rapidly (e.g. with high scan rate) all masses emerging from the mobility spectrometer. The combination of a mobility spectrometer with an oTOFMS is referred to as an Ion Mobility-oTOFMS. This prior art instrument comprised means for ion generation, a mobility drift cell, an oTOFMS, and a small orifice for ion transmission from the mobility cell to the oTOFMS.
In 2003, Loboda (U.S. Pat. No. 6,630,662) described a method for improving ion mobility separation by balancing ion drift motions provided by the influence of DC electric field and counter-flow of the gas. Using this balance, ions are at first accumulated inside an ion guide, preferably an RF-ion guide, and then, by changing the electric field or gas flow, the ions are gradually eluted from the ion guide to the mass spectrometer. Such type of ion accumulation is restricted to collecting relatively small number of ions due to space-charge effect. It also has some limitation in ion mass-to-charge (m/z) range because RF-focusing for a given RF-voltage has decreasing efficiency for larger mass ions. Increasing RF-voltage in this case is limited due to the possibility of glow discharge at high voltages. For at least these reasons, this method has significant resolving power limitations, particularly for large mass ions. The time of ion accumulation and their storage in RF-ion guide should not be too long, otherwise ions would be partially lost due to diffusion into rods or walls confining the gas flow. The instrumental improvements disclosed below eliminate these drawbacks.
Use of MS as a detector enables separation based on mass-to-charge (m/z) ratio after the separation based on ion mobility. Shoff and Harden pioneered the use of Mobility-MS in a mode similar to tandem mass spectrometry (MS/MS). In this mode, the mobility spectrometer is used to isolate a parent ion and the mass spectrometer is used for the analysis of fragment ions (also called daughter ions), which are produced by fragmentation of parent ions. Below this specific technique of operating a Mobility-MS is referred to as Mobility/MS, or as Mobility-TOF if the mass spectrometer is a TOFMS-type instrument. Other prior art instruments and methods using sequential IMS/MS analysis have been described (see, e.g., McKight, et al. Phys. Rev., 1967, 164, 62; Young, et al., J. Chem. Phys., 1970, 53, 4295; U.S. Pat. Nos. 5,905,258 and 6,323,482 of Clemmer et al.; PCT WO 00/08456 of Guevremont) but none combine the instrumental improvements disclosed here. When coupled with soft ionization techniques and the sensitivity improvements obtained through the use of the drift cell systems disclosed herein, the IMS/MS systems and corresponding analytical methods of the present invention offer significant analytical advantages over the prior art, particularly for the analysis of macromolecular species, such as biomolecules.
One challenge when building a Mobility-MS system is to achieve high ion transmission from the mobility region into the MS region. It is at this interface that earlier uses of linear fields appear incongruous with the goal of maximizing ion throughput across the IMS/MS interface. The mobility section operates at typical pressures between 1 mTorr and 1000 Torr whereas the MS typically operates at pressures below 10−4 Torr. In order to maintain this difference in pressure it is necessary to restrict the cross-section of the exit orifice of the IM drift cell so that the region between the IM and the MS can be differentially pumped. Typically this orifice cross section is well below 1 mm2. Hence it is desirable to focus the ions into a narrow beam before they reach the interface. Another important property of ion beam coming into MS is the beam divergence, or the kinetic energy of ion motion in the plane orthogonal to the direction of their travel. This is the main factor responsible for the quality of mass spectra obtained in the orthogonal TOFMS. It is a subject of the present invention to achieve good ion beam properties by using a thin dielectric coating of the electrodes followed by controlled charging of this coating. It allows the use of a channel instead of an exit orifice with sharp edges for the IM drift cell and to form low divergent supersonic gas flow where ions could be significantly cooled to have an average energy of their side motion corresponding to a few ° K.
In 1997, Brittain, et al. (U.S. Pat. No. 5,633,497) described the coating of the interior surfaces of an ion trap or ionization chamber with an inert inorganic non-metallic insulator or semiconductor material for the passivation of the surfaces in order to minimize absorption, degradation or decomposition of a sample in contact with the surface
U.S. Pat. No. 6,600,155 to Andrien et al., teaches the coating of a surface in time-of-flight pulsing region with a dielectric film (among other types of films) for improving ion beam properties before orthogonal extraction of ions into the drift region of a time-of-flight mass spectrometer
Whitehouse (U.S. Pat. No. 6,707,037) proposed the extraction of ions of both signs from a MALDI target directly located inside gas-filled RF-multi-pole ion guide, to concentrate them along the axis of the guide, and send them in opposite directions under the influence of an axial electric field for subsequent mass analysis.
Park (US Patent Application No. 2004/0149902 A1) proposed the use of a multi-pole RF-ion guide to insert ions from a number of ion sources into analytical devices including mass spectrometers and mobility spectrometers. In 2002 Moini and Jiang in U.S. Pat. No. 6,465,776 described the insertion of ions from multiple electrospray capillaries through one quadrupole RF-ion guide where ion beams are mixed into TOFMS. However, multi-channel data recording was not disclosed.
U.S. Pat. No. 5,763,865 to Kaersdorf et al. disclosed a method and apparatus for quantitative non-resonant photoionization of neutrals. A time-of-flight mass spectrometer with novel ion mirror for separation of different ion beams is described. Eriksson, in U.S. Pat. No. 6,683,302 described an electrospray ion source wherein heating of droplets emerging from the electrospray capillary under the influence of strong electric field is provided by a microwave field between the spray tip and mass analyzer. U.S. Patent Application No. 20030226750 of Fenn suggests the use of AC voltage to produce charged droplets from the solution emerging from a conducting capillary instead of DC voltage used in conventional electro-spray ion sources. It was disclosed that the flow of the droplets for 60 Hz 5 kV AC voltage is similar to that of a conventional electrospray (ESI) ion source. In 2003 Ranasinghe et al. (US Patent Application No. 2003/0001090) proposed to split the liquid flow from some separation device into two approximately equal streams and direct them into two ion spray sources—the first one producing positive ions and the second one producing negative ions. Two TOFMS systems are used to record positive and negative ions separately.
In 2002, Berggren, et al. (US Patent Application No. 2002/0166961) described a charged droplet source for mass spectrometer with the focusing of droplets and ions using an aerodynamic lens. This lens is a sequence of coaxial apertures where the gas flow comes through orifices with decreasing diameter so that charged particles are focused to some extent.
In 2003 Cornish et al. (U.S. Pat. No. 6,580,070) suggested to combine several relatively simple coaxial TOFMS systems with MALDI or laser ablation ion sources. One array located inside a vacuum chamber provides high throughput analysis of several samples or one large sample in different points. In 2004 Hobbs, et al. (US Patent application No. 20040217279) described multianalyzer mass-spectrometer for the parallel analysis of multiple samples preferably coupled with fluid phase separation techniques
All of the above-referenced U.S. patents and published U.S. patent applications are incorporated by reference as though fully described herein.
Although much of the prior art resulted in improvements in ion focusing, separation and in ion throughput from ion source to the mobility cell and to the mass spectrometer in tandem instruments, there is room for additional improvement in all these directions. The inventors describe herein a concept and designs of multi-beam ion mobility and mass separations with multi-channel data recording which result in variety of instrumental embodiments to provide improved ion production from investigated samples, their separation and measurements.
The present invention is directed to a system and method for mass spectrometric analysis of samples. The present invention mainly deals with systems and methods using ion mobility drift cells for transporting ions through a high pressure gas to a TOF MS. The following concepts are described in various embodiments of the present invention: (i) multi-beam ion pre-selection from a single sample, (ii) coordinated mobility and mass separation of ions by single or plural TOFMS (independently for different beams) with controllable, on-demand ion fragmentation, and (iii) multi-channel data recording to increase the efficiency of sample use in the analysis or to obtain as much useful information as possible about the sample in a reasonably short time. Specifically, the improvements lie in (i) extracting or producing both positive and negative ions from the sample (ii) accumulation and post-ionization of neutrals, (iii) trapping of ions in a plurality of traps, (iv) improved ion transportation against the gas flow into mobility cells, and (v) a new design for a mobility cell/TOFMS interface comprising well collimated supersonic cooled gas flows into a unique multi-channel RF-ion guide. In comparison to conventional methods, improvement (v) provides significantly lower divergence of the ion beam entering the TOF MS which in turn improves the mass resolution and mass accuracy of the TOFMS measurement. Because of this new approach, the divergence of the ion beam is low enough to use a simple linear TOFMS instead of more complicated reflectron type TOFMS and still obtain mass resolution in the range of several thousands. It is possible also to decrease further the divergence of the ion beam by using parabolic (or quasi-parabolic) ion mirror made of conductor coated by a variety of non-conducting (or poorly conducting) thin films. Parabolic mirrors include parabolic mirrors, cylindrical parabolic mirrors, and other parabolic variations. It is possible in this case to achieve resolving powers of more than 10000 for small (˜20 cm length) linear time-of-flight mass spectrometer. The multi-channel data recording system not only acquires separate data from each ion mobility channel but also provides an increased dynamic range of for ion intensity measurements within the TOFMS. It also allows the determination of additional characteristics of ions such as velocity and divergence, which in conditions close to equilibrium with gas flow, provides a direct estimation of the charge state. These advantages of multi-channel data recording may also be implemented in a simple way in a linear TOFMS. These improvements may be used to increase throughput from an ion source to downstream instruments and they may also provide additional information about the samples beyond merely summing of the data for different ion beams. The resulting instruments and methods are useful for qualitative and/or quantitative chemical and biological analysis.
In the present invention it is taught to accumulate both negative and positive ions from MALDI target or from other sources (secondary ion sources, fast atom bombardment sources, electrospray source, atmospheric pressure ionization sources) in RF multi-pole ion traps, which gives a possibility to collect ions from the target while the previous portion of ions is moving and being recorded. It may give better and more effective conditions for producing and collecting ions and thus higher sensitivity. After ion accumulation the operation of the RF-multi-pole ion traps is altered so that ions of both sign ions can be inserted against the buffer gas flow into the two opposing arrays of multi-channel ion mobility cells. After the insertion of the largest desired ions into the entrance plane of a desired ion mobility channel, the cycle of ion accumulation is repeated once again. The main advantage of such ion insertion against the gas flow compared to prior art is the significant increase of mobility resolution and better control over the time of mobility separation.
In one aspect of the present invention, there is an apparatus for analyzing a flow of gaseous ions or mixture of gaseous ions and gaseous neutral species, the apparatus comprising an ion mobility assembly comprising an ion trapping region comprising a plurality of ion traps to receive the flow of gaseous ions or mixture of gaseous ions and gaseous neutral species; a plurality of parallel mobility tubes, each of the mobility tubes fluidly coupled to the ion trapping region; optionally, a plurality of parallel CID tubes, each of the CID tubes fluidly coupled to at least one of the mobility tubes; optionally, a plurality of parallel exit tubes, each of the exit tubes comprising at least one electrode and being fluidly coupled to at least one of the CID tubes or to at least one of the mobility tubes; and, at least one multi-channel RF ion guide fluidly coupled to at least one of the exit tubes, at least one of the CID tubes, or at least one of the mobility tubes; the ion mobility assembly having a separation axis in a first direction; and, at least one TOFMS fluidly coupled to the ion mobility assembly, the TOFMS comprising a position sensitive detector. In some embodiments, the ion trapping region is operable under a pressure of about 0.1 to about 10 Torr. In some embodiments, the plurality of ion traps comprises are least one RF ion trap. In some embodiments, the plurality of ion traps comprises at least one DC field trap. In some embodiments, the apparatus further comprises voltage grids between the ion traps and the first mobility tubes. In some embodiments, the apparatus further comprises at least one collimating electrode between the ion traps and the first mobility tubes. In some embodiments of the apparatus, the mobility tubes comprise a front element operable at variable voltage biases. In some embodiments, the front element comprises a discrete section of capillaries, a microchannel plate section, or a combination thereof. In some embodiments, the one or more of the mobility tubes and/or CID tubes comprises a surface which is at least partially coated with a thin dielectric film. In some embodiments, the at least one of the multi-channel RF ion guides comprises multiple pairs of rods and confining plates. In some embodiments, the apparatus further comprises at least one ion mirror between the at least one multi-channel RF ion guide and the at least one TOFMS. In some embodiments comprising at least one mirror, the at least one ion mirror comprises a conductor coated by a dielectric film. In some embodiments comprising at least one mirror comprising a conductor coated by a dielectric film, the at least one ion mirror comprises a parabolic mirror, a cylindrical parabolic mirror, or a quasi-parabolic mirror; and, a flat mirror. In some embodiments comprising at least one mirror comprising a conductor coated by a dielectric film, the ion mirror comprises a surface that is charged. In some embodiments comprising at least one mirror comprising a conductor coated by a dielectric film wherein the ion mirrors comprise thin metallic strips, the strips being alternately biased by positive and negative voltages, the voltages being RF voltages or DC voltages. In some embodiments comprising at least one mirror comprising a conductor coated by a dielectric film, the ion mirrors comprise electrode strips on a piezoelectric film surface. In some embodiments of the apparatus, the apparatus further comprises an ion interface assembly located at a high pressure/high vacuum interface between the ion mobility assembly and the at least one TOFMS. In some embodiments wherein the apparatus further comprises an ion interface assembly located at a high pressure/high vacuum interface between the ion mobility assembly and the at least one TOFMS, the ion interface assembly comprises an entry electrode, one or more focusing electrode assemblies, at least one ion mirror with a dielectric coating, a sectioned tube coated with a dielectric film, and a field-free tube coated with a dielectric film and having a larger diameter than the sectioned tube. In some embodiments of the apparatus, the TOFMS comprises a multi-channel detector. In some embodiments, the TOFMS is an oTOFMS. In some embodiments, the TOFMS is a LoTOFMS. In some embodiments, the TOFMS is a gridless LoTOFMS. In some embodiments, the ion mobility assembly comprises more than one ion mobility assembly and wherein one or more pairs of the more than one ion mobility assembly are opposed pairs. In some embodiments, the apparatus further comprises repeller electrodes divided into strips, the repeller electrodes located between the ion traps and the mobility tubes. In some embodiments, the apparatus further comprises an orthogonal collection region. In some embodiments comprising an orthogonal collection region, the orthogonal collection region comprises one or more voltage grids. In some embodiments, the apparatus further comprises a source for the generation of the flow of gaseous ions or mixture of gaseous ions and gaseous neutral species, the source being fluidly coupled to the ion mobility assembly, and the source being configured to generate the flow in a direction that is orthogonal to the first direction. In some embodiments of that apparatus having an orthogonal source and ion mobility assembly, the source further comprises means for post-ionization of gaseous ions or gaseous mixtures of ions. In some embodiments of that apparatus having an orthogonal source and ion mobility assembly, the source is selected from the group consisting of a laser desorption source, a cluster bombardment source, a secondary ion source, an electrospray ionization source, photoionization source, a pneumo-spray source, an atmospheric pressure ionization source, and any combination thereof. In some embodiments wherein the source is a laser desorption source, the laser desorption source is a matrix assisted laser desorption ionization source. In some embodiments of the apparatus wherein a source is present, the source is a secondary ion source. In some embodiments employing a secondary ion source, the apparatus further comprises an electrically conducting input tube having an entrance region and an exit region, the entrance region fluidly coupled to the source and the exit region fluidly coupled to the ion trapping region. In some embodiments employing a secondary ion source and further comprising an electrically conducting input tube having an entrance region and an exit region with the entrance region fluidly coupled to the source and the exit region fluidly coupled to the ion trapping region, the electrically conducting input tube has an internal surface comprising a dielectric film coating. In some embodiments employing a secondary ion source and further comprising an electrically conducting input tube having an entrance region and an exit region with the entrance region fluidly coupled to the source and the exit region fluidly coupled to the ion trapping region, the ion trapping region comprises a plurality of electrode triads and a plurality of RF quadrupoles. In some embodiments employing a secondary ion source, the apparatus further comprises a laser source positioned to provide laser radiation to the ion trapping region.
In one aspect of the present invention, there is a method of analyzing a sample comprising the steps of creating a flow of gaseous ions or mixture of gaseous ions and gaseous neutral species from the sample, the flow having an axis of flow in a first direction; injecting the flow into an ion mobility assembly, the ion mobility assembly comprising: an ion trapping region comprising a plurality of ion traps to receive the flow; a plurality of parallel mobility tubes, each of the mobility tubes fluidly coupled to at least one of the ion traps; optionally, a plurality of parallel CID tubes, each of the CID tubes fluidly coupled to at least one of the mobility tubes; optionally, a plurality of parallel exit tubes, each of the exit tubes comprising at least one electrode and being fluidly coupled to at least one of the CID tubes or to at least one of the mobility tubes; and, at least one multi-channel RF ion guide fluidly coupled to at least one of the parallel exit tubes, at least one of the CID tubes, or at least one of the mobility tubes; the ion mobility assembly having a separation axis that is orthogonal to the first direction; and, thereafter injecting the flow into at least one TOFMS fluidly coupled to the ion mobility assembly, the TOFMS comprising a position sensitive detector. In some embodiments, the step of creating comprises the step of applying a collimated stream of gas to the flow. In some embodiments, the method further comprises the step of maintaining a pressure of about 0.1 to about 10 Torr in the ion trapping region. In some embodiments, the method further comprises the step of applying a DC electric field to the ion trapping region. In some embodiments, the method further comprises the step of applying an RF voltage to the ion trapping region. In some embodiments, the step of creating further comprises the step of applying an RF voltage to the flow before the step of injecting the flow through the ion mobility assembly, wherein the RF voltage applied to the flow before the step of injecting through the ion mobility assembly is shifted by π/2 with respect to the RF voltage applied in the ion trapping region. In some embodiments, the method further comprises the step of cooling the flow before the step of injecting the flow through the at least one TOFMS. In some embodiments of the method, the step of cooling comprises cooling with supersonic gas flows. In some embodiments of the method, the step of creating comprises extracting and ionizing gaseous neutral species from a gaseous sample plume. In some embodiments of the method wherein the step of creating comprises extracting and ionizing gaseous neutral species from a gaseous sample plume, the said step of extracting comprises passing a collimated stream of gas through the gaseous sample plume. In some embodiments of the method wherein the step of creating comprises extracting and ionizing gaseous neutral species from a gaseous sample plume, the step of ionizing comprises ionizing with laser radiation. In some embodiments of the method wherein the step of creating comprises extracting and ionizing gaseous neutral species from a gaseous sample plume, the step of ionizing comprises fragmentation of neutral zwitterions. In some embodiments of the method, one or more of the plurality of parallel mobility tubes comprise a front element and the method further comprises the step of applying a constant or variable voltage bias to the front element. In some embodiments of the method, the at least one multi-channel RF ion guide comprises one or more multi-pole RF ion guides. In some embodiments wherein the at least one multi-channel RF ion guide comprises one or more multi-pole RF ion guides, the multi-pole RF ion guides comprises two or more rods and the method further comprises the step of applying a voltage of varying phase and amplitude to the rods. In some embodiments of the method, the step of creating the flow comprises creating droplets with a component selected from the group consisting of an electrospray source, a pneumo-spray source, an atmospheric pressure ionization source, and any combination thereof. In some embodiments of the method comprising the step of creating droplets, the method further comprises the step of splitting the droplets into positively and negatively charged droplets by a quasi-resonant sound frequency field or an ultrasound frequency electric field. In some embodiments of the method comprising the step of creating droplets, the method further comprises the step of drying the droplets by ambient gas heating, microwave absorption, or a combination thereof. In some embodiments of the method comprising the step of creating droplets, the method further comprises the step of applying an electric field to the droplets. In some embodiments of the method, the step of creating the flow comprises creating the flow with a component selected from the group consisting of a laser desorption source, a cluster bombardment source, a secondary ion source, an electrospray ionization source, photoionization source, and any combination thereof. In some embodiments of the method, the method further comprises passing said flow through a differentially pumped low pressure region between the ion mobility assembly and the TOFMS. In some embodiments of the method, the step of injecting the flow through an ion mobility assembly comprises injecting the flow through more than one ion mobility assembly and wherein one or more pairs of the more than one ion mobility assembly are opposed pairs. In some embodiments of the method, the step of injecting the flow through at least one TOFMS comprises injecting the flow into at least one TOFMS comprising a multi-channel detector. In some embodiments of the method, the at least one TOFMS is at least one oTOFMS. In some embodiments, the at least one TOFMS is at least one LoTOFMS. In some embodiments wherein the at least one TOFMS is at least one LoTOFMS, the at least one LoTOFMS is at least one gridless LoTOFMS. In some embodiments of the method, the step of creating comprises creating with a secondary ion source. In some embodiments of the method comprising creating the flow of gaseous ions or mixture of gaseous ions and gaseous neutral species with a secondary ion source, the method further comprises the step of passing the flow through an electrically conducting input tube having an entrance region and an exit region prior to the step of injecting the beams through said ion mobility assembly In some embodiments of the method, the ion trapping region comprises a plurality of electrode triads and a plurality of RF quadrupoles. In some embodiments of the method, the method further comprises the step of post-ionizing neutral species in the ion trapping region with laser radiation.
The foregoing has outlined rather broadly the features and technical advantages of the present invention in order that the detailed description of the invention that follows may be better understood. Additional features and advantages of the invention will be described hereinafter which form the subject of the claims of the invention. It should be appreciated by those skilled in the art that the conception and specific embodiment disclosed may be readily utilized as a basis for modifying or designing other structures for carrying out the same purposes of the present invention. It should also be realized by those skilled in the art that such equivalent constructions do not depart from the spirit and scope of the invention as set forth in the appended claims. The novel features which are believed to be characteristic of the invention, both as to its organization and method of operation, together with further objects and advantages will be better understood from the following description when considered in connection with the accompanying figures. It is to be expressly understood, however, that each of the figures is provided for the purpose of illustration and description only and is not intended as a definition of the limits of the present invention.
For a more complete understanding of the present invention, reference is now made to the following descriptions taken in conjunction with the accompanying drawing, in which:
As used herein, “a” or “an” means one or more, unless otherwise expressly indicated or obvious from the context. This is particularly true when reference is made to instrumental apparatuses or individual components of the same.
As used herein, a “plurality” means two or more (i.e., more than one).
As used herein, an “electrode triad” is a distinct group or cluster of three electrodes.
As used herein, a “mobility cell assembly” is defined as a single or multi-channel device which performs mobility separation of ions and comprises a first mobility tube, collision induced dissociation (CID) tubes wherein collision-induced ionization occurs, and final ion transport with cooling gas flow through “exit tubes” into multi-channel RF-ion guide. In the multi-channel embodiment, the mobility cell assembly comprises a plurality of first mobility tubes, CID tubes, exit tubes and RF-ion guides, preferably with each of the aforementioned component in series with one another and each series in parallel with at least one other series.
As used herein, “mobility tube” is an ion mobility cell or other cell for the transport of ions; the terms ion mobility cell and mobility tube are synonymous herein.
As used herein “collision induced dissociation tube” or “CID tube” is a tube in which high electric fields may be created sufficient to provide collision-induced dissociation of ions. In the present invention, the CID tube, when present can be used for collision-induced dissociation or alternatively, the collision-induced dissociation mode may be disabled and the CID tube may be used for cooling such as gas cooling and/or RF cooling.
As used herein “exit tube” is a tube with a diameter less than a preceding tube to which it is fluidly coupled, said exit tube being used as a gas collimating and ion transport tube.
As used herein, the term “separation axis” as it relates to an ion mobility assembly or any individual component of an ion mobility assembly is the axis defining the direction of travel of ions and/or neutral species traversing or being transported through the ion mobility assembly or any individual component of the ion mobility assembly.
As used herein, a “ion trapping region” or “trapping region” is part of the entrance interface between the ion source and the mobility cell where ions preferably of both signs and possibly neutrals are accumulated. Neutrals are ionized and the resulting ions are trapped in multiple traps before penetrating a mobility cell.
As used herein, an “orthogonal collection region” is defined by the volume between at least one electrode and/or voltage grid pair through which ions (possibly of both signs) and neutrals which are mixed with a carrier gas pass orthogonally in front of the entrance of at least one IM tube. Neutrals which are formed within the gas flow through this region are transformed into ions by an ionization or fragmentation process (such as by a laser) within this region.
As used herein, “MALDI” means matrix assisted laser desorption ionization.
As used herein, “SIMS” means secondary ion mass spectrometry.
As used herein, “FAB” means fast atom bombardment mass spectrometry.
As used herein, the term “TOFMS” is defined as a time-of-flight mass spectrometer including both the linear or reflectron type; as used herein, “oTOFMS” is defined as a time-of-flight mass spectrometer both linear or reflectron type configured orthogonally to the analytical axis of a preceding instrumental platform such as, for example, the separation axis of an ion mobility cell; “LoTOFMS” is specifically defined as a linear oTOFMS. The oTOFMS and the LoTOFMS are examples of a TOFMS.
As used herein IM-oTOFMS refers to a combination of an Ion mobility spectrometer with an orthogonal time of flight mass spectrometer.
In applications which use ion mobility cells filled with a few Torr of buffer gas as a volume/charge separation stage in front of a mass spectrometer, the cooled ions exit through a small aperture into a differentially pumped low pressure region before the high vacuum region of the mass spectrometer. To minimize transmission losses through the small aperture, the ion beam inside the mobility cell should be focused. Ion beams should be as narrow and parallel as possible in the region between the mobility cell and TOFMS to allow the use of small differential pumping apertures (enabling lower gas flow) and to achieve higher mass resolution for TOFMS operation. Therefore the beam should be cooled as much as possible to obtain low divergence. If this divergence is small in directions orthogonal to the direction of the initial ion beam, then multiple ion beams may be kept isolated all the way from the ion source to the multichannel detector plane in the TOFMS in order to increase the instrument throughput (proportional to the number of ion beams). Such an approach is feasible because: (i) devices for multi-channel data recording (multi-channel time-to-digital converter (“TDC”)) are available and widely used; (ii) it is possible to transport ions inside small tubes without losses by coating conducting surfaces with thin dielectric films as described in details below; (iii) it is possible to organize mobility separation of ions in a gas counter-flow in a controllable time scale so that the use of different channels of TDC for position sensitive detection may be used for recording of plurality of ion beams; (iv) gas flow through narrow tubes from high pressure to vacuum can have divergence corresponding to a gas temperature of 1° K or less; and (v) ions hundreds of times heavier than gas atoms and traveling in such a gas flow may have a divergence corresponding to about 10° K temperature.
In the present invention, a multichannel IM-oTOFMS detection unit (400) shown schematically in
It should be noted that the instrument of
The ions may be transported to one or more intervening mobility tubes in series with an exit tube. At certain time intervals high electric field may be created inside or between sections to provide CID (collision induced dissociation) of specific ions. Alternatively, the front section of the exit tube element (16) can either be used for CID or can be used for cooling applications without collision induced dissociation. After passing through the middle region of mobility cell (5) ions penetrate the region of exit tubes (7). The exit tube (16) diameter is less than the preceding tube (15) to which it is fluidly coupled and is used as a gas collimating tube. The mobility carrier gas flows through each exit tube (16) along with mobility separated ions. The exit tubes (16) must also have electrodes for creating electric fields which will move ions since the gas flow at the beginning of these tubes is relatively slow. By choosing and controlling the pressures in region (7) and region (8) it is possible to create a highly directed gas beam which emerges from the exit tubes into vacuum with low divergence (corresponding to the absolute temperature of about 1K or lower). Ions moving with gas in these tubes are only partially cooled since they do not have quite enough time to come close to equilibrium with the helium gas flow, so their divergence is somewhat higher. It is at this point that a multi-channel RF-ion guide (8) is used to focus and cool each ion beam (123) so that the ions attain the velocity close to that of cooled IM buffer gas beam (Helium in the example). This ion guide (8) comprises multiple pairs of rods (18) (in the horizontal plane) each pair of which is separated by confining plates (17) (in the vertical plane). RF-voltage of the same phase is applied to the rods (18). Confining plates (17) have the same voltage as oTOFMS (10). These plates provide focusing of ions between rods of each pair. This apparatus provides low divergent ion beams which may be inserted through small orifices (19) directly into an oTOFMS or alternatively (as shown in
Reference to
In the preferred embodiment of the present invention, charged thin film coatings of conducting surfaces are used to increase the mobility cell ion transmission. This coating was described in U.S. Pat. No. 6,992,284 to Schultz, et. al. where the coating is used to increase the mobility cell ion transmission. The result of computer calculation of potential distribution near such coated surface is shown in
In another embodiment of the present invention, there is a significant cooling of gas emerging from the tube connecting regions of differential pressures. Corresponding measurements were performed using experimental setup which is shown in
Data shown in
One embodiment of the present invention enables extraction of ions of both signs from several spots on a MALDI sample. This is shown in
Another way to maximize sample use is to collect neutrals from the sample plume and generate ions in addition to the ions initially formed in the MALDI plume. This is illustrated in
It seems at first that it is only necessary to measure positive ions created from neutrals since formation of negative ions during laser ionization is expected usually only by subsequent secondary electron attachment reactions. However, direct formation of ion pairs can occur, especially for the case of biomolecules which may exist in a preformed state within the sample in the form of zwitterions with equal number of positive and negative charges. Breaking bonds at different sites may produce such distinct positive and negative ion pairs giving valuable structure information. The energy of about 2 eV may be sufficient to break a peptide bond which thus can, depending on the particular structure of the zwitterion, create two separated ions of opposite sign. (Note, this can also be used to analyze the structure of ions where the net sum of negative and positive ions on the molecule differ by one or more charges). For direct ionization of organic molecule the energy of about 10 eV is necessary although this direct ionization by one or more photons often is accompanied by significant molecular fragmentation. A sequence where initially the accumulation of neutrals along the axis of helium flow through the trapping region based on their size related diffusion coefficient within the gas flow may be important for analysis of ions subsequently produced from these neutrals. Two ions of opposite sign which come from the zwitterions of some type should be correlated in the distributions for the ion beams (ideally should belong to the pair of corresponding ion beams) when simultaneously measured. This criterion, besides the assumption that each positive ion from a zwitterion should have the corresponding negative ion and sum of their masses should be equal to the mass of the zwitterion, may provide valuable information about the structure and sequence of these zwitterions. This could be important since often this information is difficult to obtain even when using CID or similar MS/MS techniques. Some difference in co-incident intensities of simultaneously produced ions from the zwitterion is possible due to different efficiency of ion trapping for negative and positive ions. Also secondary processes which further transform one or the other of the photofragment daughter ions are possible as well. Usually biomolecular zwitterions in the sample of natural origin have a distribution of charges and are not a single type of ions because they retain charges in different sites of the biomolecule according to a stochastic processes. So if a pair of positive and negative ions is suspected to belong to a zwitterion it is highly probable that positive and/or negative ions of the same mass (corrected for the number of attached or lost protons or, perhaps, alkali metals) would be recorded among ions coming directly from the MALDI target. Similar types of photo-fragmentation and coincidence between positive and negative structural fragments would also be expected from fragmentation of either the positive or negative ions when such ions had one or the other charge in excess in a structure which was charged at multiple locations. Thus if an ion which was for example nominally singly charged but contained a total of two positively charged sites and one negative, then the resulting fragmentation products could be either two “coincident” ions (one negative and one doubly charged positive), or one positive and one neutral fragment depending on whether the cleavage point was between the two positive charge locations or between the positive and negative charge location. Using four measuring units (81, 85, 88 and 90) provides better conditions for accumulation of neutrals around axis (83) because gas flows (89) from these units would force these neutrals to the axis from all directions. Furthermore, in addition to ions and neutral molecular species the MALDI plume can contain droplets or some large associations of molecules and ions. In such cases when the size of such particles is fairly large (1 μm or more), they cannot obtain significant velocity in the helium flow on such a short distance (about 2 cm). They may come to the right wall of the trapping region and can be accumulated there without evaporation of the solvent or significant degradation. Therefore it is reasonable to heat this wall to a temperature of about 100° C. or higher to evaporate the solvent from these droplets and release the neutrals or ions contained within. Ions and neutrals produced in such a way would be trapped in RF-traps or near the axis. It is probable in this case that ions would be trapped in the last trap (at the right of the trapping region) independently of their sizes. Detecting ions from this trap with and without wall heating would indicate the mechanism of droplet ion formation. It may also be reasonable to control the initial temperature of the gas (helium) to produce a fast narrow gas flow. In this case the divergence of the flow would not change much but the gas flow velocity would increase as thermal energy of the gas in this flow is transformed almost entirely into kinetic energy of axial motion of gas atoms. Droplets from MALDI target will be heated by the gas and may evaporate without freezing. This may produce additional ions and neutrals ready for ionization and thus enhance the sensitivity of measurements. The intensity and divergence of helium flow may be controlled by adjusting Pflow and Ppump—FIG. 8—(changing effective pumping rate) as well. Therefore, the neutral and ion trapping abilities and the distributions of trapped neutrals and ions along the axis can be varied. Switching off the helium flow removes all neutrals from the trapping region.
A possible way of re-trapping and gradually inserting ions into a multichannel structure (which might be for example microchannel plates used as mobility tubes) is shown in
Helium flow (6) introduction and collision induced dissociation of specific ions can be performed in the set of CID tubes (
Multi-Channel Exit Interface
The separated ion beams as shown in
The RF field variation would also give an opportunity to effectively focus large mass ions to the same beam width as smaller ones. This is true for the singly charged ions; moreover multi-charged ions will be focused even better—proportionally to the charge. When the CID tubes are used to form CID products, it is important to point out the following. CID is usually used to obtain structural information about ions and the most valuable information is obtained from fragments with masses close to that of the parent ion. Ions moving with gas having 1K temperature and focusing in RF-ion guide come closer and closer to thermal equilibrium with the gas and their temperature becomes low. Since the width of an ion beam focused in a RF-ion guide is proportional to the square root of the ion temperature, the ion beam width may significantly decrease. For example, ions having 3K temperature would have the final beam width 10 times smaller than that of ions at room temperature (˜300K). It is possible to obtain ion beams much less than 1 mm in diameter for room temperature RF-quadrupoles, the beam width of low temperature ions may be less than 0.1 mm. If input orifices in TOFMS (130) can be about 0.1 mm in diameter, low gas flow (122) will penetrate inside TOFMS and the desirable low gas pressure inside TOFMS may be provided by using relatively modest pumping power.
oTOFMS Suitable for Multi-Beam Ion Recording after Ion Cooling in the Exit Interface
If ions coming from the exit interface (
The fact that the velocity distribution of ions is made equal to the velocity of the He by the RF cooling in region 8 gives virtually the same velocity to all ions over a wide mass range. This narrow, mass independent velocity distribution allows better use of the ion accumulation time in the extracting region. The problem usually encountered in orthogonal TOFMS is that ions with medium to larger masses are detected with better efficiency than lower mass ions simply because the low mass ions (when compared to the slower velocity large ions) move quickly through the orthogonal extraction region and are mostly lost. Also, losses of really high masses falls off in part because these ions are moving so slowly that it takes a very long time to fill the orthogonal plates and, therefore, a large fraction of these larger masses do not have time to adequately fill the region between the orthogonal extraction plates and are lost at the front region of the extractor. On the other hand, a potential problem with the present invention is that because the flight time through the analyzer is correlated to the ion velocity in the initial direction of ion motion (in case of ion acceleration by electric field) the reflected ions with axial velocity close to the drift velocity of the gas may partially or entirely miss a detector which is located coaxially with the extractor plates. To overcome this drawback it is possible to tilt the ion mirror with respect to the ion flow/extraction plates and place the detector parallel to the extraction as it is suggested, for example, in the U.S. Pat. No. 6,683,299, K. Fuhrer et. al., 2004, incorporated by reference as though fully described herein.
Another way to avoid missing the detector is to use a Linear TOFMS (right-bottom part of
To decrease the “dead” time for inserting ions into the acceleration region (131) of
A grid free operation of the instrument (
Reduced pumping requirements for achieving high vacuum inside the LoTOFMS (124) is a further advantage of this arrangement shown in
Measurement of Beam Divergence Using Multianode Data Recording
Ion beams coming into the TOFMS have the width of about 1 mm and the average divergence of about 0.01 radian. As simulations show, the axial energy for large ions from such a gas flow is gradually increasing with their size and the beam divergence is decreasing. If the orthogonal extraction duty cycle of TOFMS is about 30 μsec then for a linear TOFMS ions fill the extraction region for 15-25 μsec and then travel in the TOFMS for less than 30 μsec. If ions have an axial velocity of about 1000 m/sec they will shift in this direction by about 5 cm and the standard deviation of the ion beam width in the plane of recording (due to 0.01 divergence) will be about 0.5 mm. If the distance between ion beams is about 2 mm these beams will overlap to some extent at the end of recording plate. Thus if the detector has eight anodes and each one is used to record an individual ion beam, it will record certain amount of signal from the adjacent beams too. This apparent drawback may be turned into an important advantage. A relatively small part of each ion beam is recorded on adjacent anodes. We can use this to increase the dynamic range of measurements when the signal in the main channel is saturated. It is formally the same concept as that of using two anodes of large and small sizes to intercept different fractions of the ion flux on the detector (see our U.S. Pat. No. 6,747,271, Gonin, et al, 2004). With good m/z and mobility separation and several adjacent anodes for ion recording the problem can be solved in many cases.
Sometimes overlapping ion peaks may prevent precise measurement of the individual intensities but this situation is quite common in mass spectrometry and may be solved either completely or at least to some extent by deconvolution and curve fitting taking into account individual peak shapes and isotopic distributions. The same mass ions coming from different channels under the same starting conditions may appear at different times on the same anodes (due to mechanical inaccuracy, for example) and the corresponding peaks may not overlap. The coefficients for recovering the saturated signal in the main channel may be obtained by comparing the signals on anodes collecting the tails of mobility peaks (i.e. where the main signal is not saturated yet). These coefficients for known location and sizes of recording anodes could be converted easily into the angle divergence of ion beams if the velocities of ions in axial direction are known. It is possible to obtain velocities by using signals from several (two or three) separate anodes. These anodes should divide the expected spread into equal parts. The signal from each part with the signals from adjacent channels would allow estimation of the average axial velocity of a given ion of as well as its angular divergence or temperature. These two values (the axial velocity and the temperature of ions for specific experimental conditions) may provide additional information about ions which does not strictly depend on their m/z ratio and mobility. Ion velocity would be mainly determined by the ion mass and its cross-section for collisions but this cross-section may be different from the mobility cross-section. Since ions move in a very cold gas, they will be significantly cooled and their conformation may change. They may become more compact and have a noticeably smaller cross-section compared to that at higher temperatures. It is possible to get some additional information about the shape of these cooled ions by measuring their ion velocity and angular divergence using only eight anodes and recording eight ion beams. The corresponding anodes should be located at a certain angle relative to the direction of ion beams as shown in
In case angular divergences of ion beams on the detector are too low to provide convenient increase of the dynamic range of the measurements it is possible to either provide more anodes and/or to produce a non-uniform focusing of the ion beams. Ion divergence in the direction of ion orthogonal extraction should be as small as possible to get high resolution whereas divergence in the orthogonal direction may be significantly large. To obtain such focusing, the multi-channel RF-ion guide rods (18), as in shown at the top of
Recording of Elemental and Molecular Ions Using SIMS
Recording of Ions and Neutrals from Spray Ion Sources (Electron, Pneumo, Laser)
Recording of Ions and Neutrals from Atmospheric Pressure Ionization Ion Source
Almost the same entrance interface may be proposed for an atmospheric pressure ionization source for direct analysis of admixtures in ambient air.
The details in the
Although the present invention and its advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims. Moreover, the scope of the present application is not intended to be limited to the particular embodiments of the process, machine, manufacture, composition of matter, means, methods and steps described in the specification. As one of ordinary skill in the art will readily appreciate from the disclosure of the present invention, processes, machines, manufacture, compositions of matter, means, methods, or steps, presently existing or later to be developed that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein may be utilized according to the present invention. Accordingly, the appended claims are intended to include within their scope such processes, machines, manufacture, compositions of matter, means, methods, or steps.
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