Ion source for inductively coupled plasma mass spectrometry

Abstract

An ICP source (100), which uses inductively coupled plasma to generate ions, is configured to be connected to a mass spectrometer (200). The sample is introduced into the plasma vertically downward (G) under the action of gravity. In this way, the sample can reach the plasma regardless of its state, e.g., droplet or particle size. Transport efficiencies up to 100% can be achieved. The ICP source can be supplied with a continuous sample-containing stream.

Description

[Technical field]

[0001] The present invention relates to an inductively coupled plasma (ICP) ion source configured for connection to a mass spectrometer. Additionally, the present invention relates to an ionization system including the ICP source, an ICP mass spectrometry system, and a corresponding method of ICP mass spectrometry. [Background technology]

[0002] Inductively Coupled Plasma Mass Spectrometry (ICP-MS) is an analytical technique that detects and quantifies trace elemental and / or isotopic ratios in a sample. This is accomplished by using an inductively coupled plasma to dissociate and ionize the sample components in an ICP source, and then separating and detecting the ions using a mass spectrometer. ICP-MS systems were first commercially introduced in the early 1980s.

[0003] ICP sources typically comprise a plasma torch with a directed gas flow to confine the plasma discharge and transport the sample aerosol to the center of the plasma discharge for dissociation and ionization. The sample aerosol is typically transported to the plasma discharge through a central injection tube of the torch. In nearly all prior art ICP-MS systems, the torch axis is oriented horizontally, whereby the sample aerosol is transported to the plasma discharge using a carrier gas that flows horizontally through a horizontal injection tube. The carrier gas flow must be carefully adjusted to optimize sample transport to the plasma discharge and ensure that the sample reaches a position within the plasma that allows it to be extracted into the mass spectrometer system with the highest possible ion yield.

[0004] Even with optimized carrier gas flow rates, the transport efficiency can be very low in prior art ICP-MS systems. This is especially true for aerosols containing relatively large objects such as large droplets, particles, or cells. In this context, it should be noted that prior art sample introduction systems for liquid samples and suspensions generally comprise a pneumatic nebulizer that provides a wide droplet size distribution, which is often followed by a nebulizer chamber that removes large droplets, e.g., droplets with a diameter larger than 10 μm, from the aerosol (see, for example, Non-Patent Document 1). With such systems, the efficiency of sample transfer into the ICP is generally in the range of 3% to 30%, depending on the nebulizer used. Certain nebulizers operating at very low liquid flow rates (less than 50 μL / min) can achieve higher efficiencies, but are prone to clogging and are therefore only useful for dilute solutions and suspensions with particles or cells in the low μm range.

[0005] Various attempts have been made to improve sample introduction into ICP-MS systems. For example, sample introduction systems using monodisperse microdroplet generators were disclosed in Non-Patent Document 2, Non-Patent Document 3, and Non-Patent Document 4. Although these documents disclose that droplets can pass vertically through a so-called drop tube, the axis of the plasma torch is still oriented horizontally, and the droplets are finally transported horizontally to the plasma using a horizontal carrier gas flow.

[0006] Non-Patent Document 5 discloses a microfluidic dispenser that introduces microdroplets into an ICP-MS. The chip is connected to a cyclone adapter, a heated steel tube, a membrane desolvator, and an ICP-MS unit. The droplets fall vertically through the steel tube, but the ICP-MS unit is oriented horizontally in the usual way, and the droplets are introduced horizontally into the ICP-MS unit.

[0007] US Patent No. 5,399,633 discloses a flow cytometer coupled to an ICP-MS system. The plasma torch of the ICP source is oriented horizontally in the usual way.

[0008] An exception to the usual horizontal orientation of the plasma torch in an ICP-MS system has been reported in [6]. The authors propose a vertically upwards plasma torch orientation. Ions from the plasma are analyzed using a vertically upwards oriented mass spectrometer with an interface connected to the plasma torch. This configuration was reported to achieve better signal stability and reproducibility for the droplet signal than prior art systems. However, the vertically upwards configuration requires very careful adjustment of the operating conditions to correctly transport the droplets to the plasma.

[0009] In the context of inductively coupled plasma optical emission spectroscopy (ICP-OES), different orientations of the plasma torch have been proposed. For example, an upwardly oriented plasma torch is disclosed in Non-Patent Document 7. The plasma torch is employed only for optical emission spectroscopy and is not compatible with mass spectrometry. Patent Documents 2, 3, and 4 also disclose upwardly oriented plasma torches, with an emphasis on applications in optical emission spectroscopy.

[0010] 8, No. 1, pp. 1111-1123, 2002, discloses a nitrogen-maintained ICP source operating at microwave frequencies and interfaced with a time-of-flight mass spectrometer. The plasma source is called a "Microwave Inductively Coupled Atmospheric Pressure Plasma (MICAP) source". It comprises an annular dielectric resonator made of technical ceramics, which, when placed in a microwave field, generates a bulk polarization current around the ring, which vibrates at microwave frequencies. The ring of the dielectric resonator has no significant net potential, and the energy coupling to the plasma is purely inductive. The plasma torch of the MICAP source is oriented horizontally. JP 2002-2003636 A discloses the MICAP source in more detail.

[0011] US Patent No. 6,399,633 discloses a microwave plasma monitoring system, in which a plasma flame is generated by microwaves in a waveguide and the light emitted from the flame is investigated by a spectrometer. In one embodiment, the plasma flame is oriented vertically downwards. Different types of samples, such as droplets or particles, are introduced using a suction pump that regulates the gas flow. The plasma source is not an ICP source and the system is designed for optical spectroscopy only and is not compatible with a mass spectrometer.

[0012] No. 5,399,633 discloses a downwardly oriented microwave plasma source for material processing, e.g. for applying surface coatings or for powder production. Downwardly oriented plasma sources for material processing are also disclosed in US Pat. No. 5,499,663, US Pat. No. 5,499,673, US Pat. No. 5,599,703, and US Pat. No. 5,599,703. These configurations are designed for industrial applications in material processing and are obviously not suitable for elemental analysis of samples. Nor are these configurations suitable for connection to a mass spectrometer.

[0013] US Patent No. 5,999,633 discloses an upwardly oriented plasma source. Droplets are delivered to a plasma torch by a single-droplet microdispenser (SDMD) with an upwardly oriented vertical orientation. The document mentions that the SDMD can also be oriented horizontally or downwardly. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] International Publication No. 2008 / 080224 [Patent Document 2] U.S. Patent No. 4,766,287 [Patent Document 3] US Patent Publication No. 2015 / 255262 [Patent Document 4] US Patent Publication No. 2016 / 0270201 [Patent Document 5] US Patent Publication No. 2016 / 0025656 [Patent Document 6] International Publication No. 97 / 13141 [Patent Document 7] US Patent Publication No. 2014 / 0287162 [Patent Document 8] US Patent Publication No. 2005 / 0211018 [Patent Document 9] US Patent Publication No. 2014 / 0217630 [Patent Document 10] US Patent Publication No. 2013 / 0270261 [Patent Document 11] US Patent Publication No. 2013 / 0118304 [Non-patent literature]

[0015] [Non-Patent Document 1] J. Olesik and L. Bates, “Characterization of aerosols produced by pneumatic nebulizers for inductively coupled plasma sample introduction: effect of liquid and gas flow rates on volume based drop size distributions”, Spectrochimica Acta Part B: Atomic Spectroscopy, 1995, 50, 285-303, DOI 10.1016 / 0584-8547(94)00151-K [Non-Patent Document 2] S. Gschwind et al., “Capabilities of inductively coupled plasma mass spectrometry for the detection of nanoparticles carried by monodisperse microdroplets”, J. Anal. At. Spectrom., 2011, 26, 1166-1174, DOI: 0.1039 / C0JA00249F Non-Patent Document 3 S. Gschwind et al., “Mass quantification of nanoparticles by single droplet calibration using inductively coupled plasma mass spectrometry”, Anal. Chem. 2013, 85, 5875-5883, DOI: 10.1021 / ac400608c Non-Patent Document 4 K. Shigeta et al., “Application of a micro-droplet generator for an ICP-sector field mass spectrometer - optimization and analytical characterization", J. Anal. At. Spectrom. 2013, 28, 646-656, DOI: 10.1039 / c2ja30207a Non-Patent Document 5 P. Verboket et al., “A New Microfluidics-Based Droplet Dispenser for ICPMS”, Analytical Chemistry 2014, 86, p. 6012, DOI: 10.1021 / ac501149a Non-Patent Document 6 II Stewart and JW Olesik, “Time resolved measurements with Single Droplet Introduction to Investigate Space-Charge Effects in Plasma Mass Spectrometry”, J. Am. Soc. Mass Spectrom. 1999, 10, 159-174 [Non-Patent Document 7] A. Murtazin, S. Groh, and K. Niemax, “Measurement of element mass distributions in particle ensembles applying ICP-OES”, J. Anal. At. Spectrom. 2010, 25, 1395-1401, DOI: 10.1039 / c004946h [Non-Patent Document 8] M. Schild et al., “Replacing the Argon ICP: Nitrogen Microwave Inductively Coupled Atmospheric-Pressure Plasma (MICAP) for Mass Spectrometry”, Anal. Chem. 2018, 90(22), 13443-13450, DOI: 10.1021 / acs.analchem.8b03251 Summary of the Invention

[0016] In a first aspect, it is an object of the present invention to provide an ICP source configured to be connected to a mass spectrometer, the ICP source enabling improved sample introduction into the plasma discharge.

[0017] This object is achieved by an ICP source according to claim 1. Further embodiments of the invention are defined in the dependent claims.

[0018] The present invention provides an ICP source for generating ions from a sample using an inductively coupled plasma. The ICP source is configured for connection to a mass spectrometer. According to the present invention, the ICP source is configured to introduce the sample vertically downward into the plasma.

[0019] For this reason, the sample introduction into the plasma is performed from the top. The sample is introduced into the plasma in the direction of the gravity vector. This makes the sample introduction less sensitive to the carrier gas flow rate. It is also conceivable to introduce the sample without any carrier gas flow at all, the trajectory of the sample before its entry into the plasma being determined only by gravity. This allows to save a large amount of carrier gas, which is quite important especially when the carrier gas is an expensive noble gas such as argon or a mixture of argon and helium. The size of the droplets or objects (e.g. cells) that can reach the plasma is practically limited only by the dimensions of the sample introduction system. Since the collision of the droplets or objects against the walls of the sample introduction system can be avoided, the proposed configuration can achieve a transport efficiency of 100%. Samples of any type and in any state can be investigated, for example aerosols or single droplets, or objects such as solid particles, cells, etc. A high degree of flexibility can be obtained with respect to sample size, aerosol humidity, and sample amount. For example, eukaryotic cells, which are typically about 10 μm to 150 μm in diameter, can reach the plasma regardless of their size or mass. Droplets reach the plasma regardless of how much solvent has evaporated during transport in the carrier gas. Sample throughput through the ICP source can be significantly increased relative to prior art devices.

[0020] Extraction of ions from the plasma can also be performed vertically downwards. To this end, the ICP source can have an extraction aperture that opens towards the bottom and is located below the region in the ICP source where the plasma is formed. After extraction, the ions can be transferred to a mass analyzer with any orientation using known ion optics such as electrostatic mirrors and / or deflection magnets.

[0021] Typically, an ICP source comprises a plasma torch, i.e. an arrangement of (usually at least three concentric) tubes that supply gas to the region of the plasma discharge. One of these tubes usually serves to supply the sample to the plasma. This tube is often called the injection tube. The injection tube is usually located centrally in the plasma torch and is surrounded by other tubes. In the present invention, the injection tube has a longitudinal axis, if any, which is oriented vertically, the upper end of the injection tube being configured to receive the sample and the lower end being configured to introduce the sample into the plasma. An extraction opening for extracting ions from the plasma can then be advantageously located below the lower end of the injection tube.

[0022] Since in the present invention the plasma torch is located above the area of ​​the plasma discharge, components such as connections supplying gas to the plasma torch can be located above the plasma discharge. To protect such components from the convection heat generated by maintaining the plasma discharge, the ICP source can include a metal cold plate that at least partially surrounds the plasma torch and thereby protects these components from the heat due to the plasma discharge. The cold plate preferably extends in a horizontal plane. The cold plate can have a cutout configured to receive the plasma torch when the cold plate is installed.

[0023] The ICP source may be equipped with a cooling system that actively cools the metal cold plate by forced flow of a fluid, e.g., a cooling gas such as air or a cooling liquid such as water, through the metal cold plate and / or through additional components connected to the metal cold plate, e.g., through one or more metal panels connecting the cold plate to a torch box surrounding the plasma torch. To this end, the cold plate and / or the components to which the cold plate is connected may be provided with one or more perforations extending into the face of the cold plate or additional components for passing a flow of cooling fluid (e.g., air or water).

[0024] An ICP source generally comprises an electromagnetic coupling element that supplies energy to the plasma by inductive coupling so as to sustain the plasma discharge. The electromagnetic coupling element may in particular be an induction coil connected to an RF generator via an electrical conductor. Such induction coils usually operate in the frequency range between 10 MHz and 100 MHz, for example at 27 MHz or 40 MHz. In another embodiment, the ICP source may employ a ring-shaped dielectric resonator placed in a microwave field, which may be generated for example by a magnetron. The microwave field may be for example in the frequency range between 1 GHz and 10 GHz, with a typical frequency of 2.45 GHz. Examples include the so-called MICAP sources disclosed in Non-Patent Document 8 and Patent Document 5, see the "Prior Art" section.

[0025] Regardless of the type of electromagnetic coupling element, the coupling element advantageously defines a longitudinal axis extending vertically. The coupling element may surround the lower end of the plasma torch. If a metal cooling plate is present, the coupling element may be disposed below the cooling plate.

[0026] The ICP source may further comprise an RF generator that provides energy to the electromagnetic coupling element. The term "RF generator" is broadly understood to encompass any AC generator operating within at least a frequency range of 10 MHz to 10 GHz.

[0027] If the electromagnetic coupling element is an induction coil, it is advantageous to connect the induction coil to the RF generator via at least one horizontally extending metal bar (preferably two parallel metal bars). This allows the induction coil to be placed at a large horizontal distance from the RF generator. The metal bar can be made of a highly conductive material, in particular copper, silver or gold, to minimize ohmic losses between the RF generator and the induction coil.

[0028] The ICP source can be combined with an aerosolization device to form an ionization system for obtaining ions from a bulk sample. Advantageously, the ICP source is connected to the aerosolization device such that the aerosolized sample is transferred vertically downward from the aerosolization device to the ICP source. This preserves the above-mentioned advantages of the vertical design of the proposed ICP source during the transfer of the aerosolized sample to the ICP source.

[0029] Depending on the type of sample, the aerosolization device can be configured in different ways. In some embodiments, the aerosolization device can include a droplet generator that aerosolizes the sample by creating droplets. A vertically oriented drop tube can be connected to the droplet generator. The drop tube is adapted to send the droplets downward under the action of gravity. As it passes through the drop tube, the droplets can reduce in size due to evaporation of the solvent or can dry out completely. To that end, the drop tube can have a sample inlet, a gas inlet, and an outlet, and the drop tube can be configured to receive droplets from the droplet generator at its sample inlet and a gas flow at its gas inlet, so that the droplets can move downward under the action of gravity while reducing in size due to evaporation of the solvent into the gas flow. The outlet of the drop tube can be directly or indirectly connected to the inlet of the ICP source such that the aerosolized sample is transferred vertically downward from the drop tube to the ICP source without the need for horizontal sample transport by a carrier gas flow as required in the prior art.

[0030] In other embodiments, the aerosolization device can include a nebulizer for aerosolizing a sample and a spray chamber connected to the nebulizer, the spray chamber having a downward facing outlet, the outlet of the spray chamber connected to an inlet of the ICP source such that the aerosolized sample is transported vertically downward from the spray chamber to the ICP source.

[0031] In yet another embodiment, the aerosolization device can include a laser ablation cell having a downward facing outlet connected to the inlet of the ICP source such that the aerosolized sample is transferred vertically downward from the laser ablation cell to the ICP source. Various configurations of the laser ablation cell are contemplated.

[0032] Other types of aerosolization devices may also be employed.

[0033] In yet other embodiments, a separate aerosolization device is not employed: instead, the injection tube of the plasma torch can constitute a direct injection nebulizer that breaks up the bulk sample liquid into droplets within the plasma torch, i.e., just before the sample enters the plasma.

[0034] The ICP source of the present invention can be combined with a flow cytometer. In general, in a flow cytometer, a sample is injected into a flow of sheath liquid. A vibrating nozzle breaks up the flow, with each droplet ideally containing one cell or particle. The flow cytometer then analyzes the optical properties of the cells or particles just before or just after the droplets are released from the nozzle. The ICP source can be connected to the flow cytometer so that the sample is transferred vertically downwards from the flow cytometer to the ICP source. This ensures maximum transport efficiency from the flow cytometer to the ICP source. Cells / particles of all sizes can fall into the ICP source and be analyzed by ICP-MS. This prevents the analysis of cells or particles from being biased towards size or mass. The flow cytometry results for each individual cell or particle can be correlated with the mass spectrometry results for that same cell or particle, maximizing the information that can be obtained from a single cell or particle. To that end, the ICP-MS system may include a correlator configured to receive flow cytometry information from the flow cytometer and mass spectrometry information from the mass spectrometer, and to correlate the flow cytometry information with the mass spectrometry information to obtain combined flow cytometry and mass spectrometry information for individual cells or particles.

[0035] Optionally, the flow cytometer can be connected to a cell sorter. In this case, objects (particularly cells or nanoparticles) that have passed through the cell sorter are transferred vertically downwards from the cell sorter to the ICP source. In particular, the cell sorter can be an electrostatic cell sorter, where droplets are given a charge depending on the result of the flow cytometry analysis. The charged droplets then fall through an electrostatic deflection system that redirects the droplets into different containers depending on the charge. Instead of collecting droplets with a particular charge state, these droplets can be directly passed to the ICP source by dropping them vertically into the inlet of the ICP source.

[0036] The flow cytometer or cell sorter can be directly connected to the ICP source. Optionally, a drop tube, as described above, can be provided between the flow cytometer or cell sorter and the ICP source to reduce the size of the droplets before they enter the plasma.

[0037] In contrast to the prior art devices, the ICP source of the present invention allows the transfer of the sample to the sample introduction system of the ICP source in the form of a continuous fluid flow. In particular, the sample introduction system of the ICP source of the present invention can be connected to any type of continuous flow liquid sample delivery system that provides a continuous flow of sample, such as a pump or a continuous flow sample preparation device. In the case of a pump, the pump can be a peristaltic pump. In the case of a sample preparation system, the preparation system can be a device for separating chemical species in a sample, such as a chromatography device or an electrophoresis device, or a device for separating populations of biological cells, such as a cell sorter connected to a flow cytometer. In contrast to the prior art devices, the ICP source of the present invention allows the transfer of the sample from the continuous liquid sample delivery system to the ICP source in the form of a continuous fluid flow, without the need for intermediate offline storage.

[0038] Generally, an ionization system is provided, the ionization system comprising: a continuous flow sample delivery system; A sample introduction system; The ICP source as described above, Equipped with the sample introduction system is connected to the continuous flow sample delivery system such that a continuous sample flow containing the sample is transferred from the continuous flow sample delivery system to the sample introduction system; The sample introduction system is configured to introduce the sample vertically downward into the ICP source.

[0039] The continuous flow sample delivery system can include a continuous flow sample preparation device having an inlet configured to receive a continuous incoming stream comprising a sample and an outlet configured to provide a continuous outgoing stream comprising the sample, at least a portion of the continuous outgoing stream comprising a continuous sample stream, and the continuous flow sample preparation device configured to perform at least one of an action on the continuous incoming stream, analyzing and / or modifying its composition.

[0040] As used herein, analysis can be performed by any known means, for example, fluorescence detection, absorption measurements, spectral analysis, etc. Similarly, alteration of the composition of the incoming sample stream can be performed by any means, such as filtration, retention by adsorption, modification by application of an electromagnetic field, etc. Importantly, no intermediate off-line storage is required between sample preparation and introduction into the ICP source.

[0041] In particular, the continuous flow sample preparation device may comprise a separation device, in particular a chromatography device, an electrophoresis device, or a cell sorter, configured to separate chemical species or particulate matter, in particular cells, in a continuous inlet stream and to elute the chemical species or particulate matter after separation into a continuous outlet stream.

[0042] Each of the inlet, outlet and sample streams may be a flow of carrier fluid carrying the sample in dispersed or resolved form. The fluid may be a liquid or a gas.

[0043] The output stream from the separation device may have a flow rate significantly greater than the maximum flow rate that the sample introduction system can accommodate. To accommodate this situation, the ionization system may include a splitter configured to continuously split the continuous output stream into a continuous sample stream and a residue stream.

[0044] The ionization system may further include a correlator configured to receive sample separation information from the separation device and mass spectrometry information from a mass spectrometer connected to the ICP source, and to correlate the sample separation information with the mass spectrometry information to obtain combined information of the separation information and the mass spectrometry information, e.g., LC / MS information.

[0045] The invention also provides an ICP-MS system comprising an ICP source of the invention and a mass spectrometer connected to the ICP source to receive ions from the ICP source, the mass spectrometer may have any orientation in space, and one or more ion optical elements may be optionally provided to deflect ions into the mass spectrometer after extraction from the ICP source.

[0046] In another aspect, the present invention provides a method of mass spectrometry comprising the steps of: maintaining a plasma using an ICP source; introducing a sample into the plasma and generating ions from the sample; analyzing the mass-to-charge spectrum of the ions extracted from the plasma using a mass spectrometer connected to the ICP source; Including, A method is provided in which the sample is introduced vertically downward into the plasma.

[0047] In a preferred embodiment, ions are similarly extracted vertically downward from the plasma to transmit the ions to a mass spectrometer.

[0048] The method is: aerosolizing the sample using an aerosolization device; transferring the aerosolized sample vertically downward from the aerosolization device to an ICP source; It may further include.

[0049] In particular, the method can include aerosolizing the sample using a droplet generator, passing the aerosolized sample vertically downward through a vertically oriented drop tube, and introducing the aerosolized sample vertically downward into an inlet of an ICP source.

[0050] In other embodiments, the method can include aerosolizing the sample with a nebulizer, passing the aerosolized sample through a nebulizer chamber having a downwardly facing outlet, and introducing the aerosolized sample vertically downward into an inlet of an ICP source.

[0051] In yet another embodiment, the method can include aerosolizing the sample using a laser ablation cell having a downward facing exit and transporting the aerosolized sample vertically downward from the exit of the laser ablation cell to the plasma.

[0052] In yet another embodiment, the method comprises: Analyzing at least one characteristic of the sample using a flow cytometer; transferring the sample vertically downward from the flow cytometer into an ICP source; may include.

[0053] The method may further include receiving flow cytometry information from the flow cytometer and mass spectrometry information from the mass spectrometer, and correlating the flow cytometry information with the mass spectrometry information to obtain combined flow cytometry and mass spectrometry information for individual cells or particles.

[0054] In yet another embodiment, the method comprises: Sorting the sample droplets according to at least one property, in particular according to an optical property, using a cell sorter; transferring a sample droplet vertically downward from the cell sorter to an ICP source; may include.

[0055] In yet another embodiment, the method comprises: providing a continuous sample flow comprising the sample from a continuous flow sample delivery system to a sample introduction system; The sample is introduced vertically downward into the plasma by a sample introduction system.

[0056] As outlined above, the continuous flow sample delivery system can include a continuous flow sample preparation device having an inlet and an outlet. The method then includes: receiving a continuous inlet flow containing a sample at an inlet; performing at least one of analyzing and / or modifying the composition of the continuous input stream; providing a continuous output stream containing the sample at an outlet; forming a continuous sample stream on at least a portion of the continuous output stream; may include.

[0057] As outlined above, the continuous flow sample preparation device may comprise a separation device, in particular a chromatography device, an electrophoresis device or a cell sorter, where the method may comprise using the separation device to separate chemical species or particulate matter, in particular cells, in a continuous incoming stream and eluting the chemical species or particulate matter after separation into a continuous outgoing stream.

[0058] The method can include continuously splitting a continuous effluent stream into a continuous sample stream and a residue stream. [Brief description of the drawings]

[0059] [Figure 1] FIG. 1 is a schematic diagram of an ICP-MS system according to an embodiment of the present invention. [Diagram 2] FIG. 2 is an enlarged view of a portion of the ICP-MS system of FIG. 1 accompanied by a highly schematic diagram of a flow cytometry-based cell sorter. [Diagram 3] FIG. 2 is a plan view of the cooling plate of the ICP source of FIG. [Figure 4]FIG. 1 illustrates a plasma torch configured as a direct injection atomizer. [Diagram 5] FIG. 1 shows a plasma torch connected to a drop tube. [Figure 6] FIG. 1 shows a plasma torch connected to a total consumption type sample introduction system. [Figure 7] FIG. 2 shows a plasma torch connected to a laser ablation cell according to a first embodiment. [Figure 8] FIG. 13 shows a plasma torch connected to a laser ablation cell according to a second embodiment. [Figure 9] FIG. 2 shows a TOF mass spectrometer suitable for connection to the ICP source of FIG. 1. [Figure 10] FIG. 2 illustrates a MICAP source according to an embodiment of the present invention. [Figure 11] FIG. 1 shows time-resolved ICP-MS peak traces of monodisperse droplets containing 133Cs. [Figure 12] FIG. 1 shows time-resolved ICP-MS peak traces of monodisperse droplets containing 232Th. [Figure 13] FIG. 13 shows a calibration curve illustrating the dependence of ion signal on analyte mass for droplets containing 133Cs. [Figure 14] FIG. 13 shows a calibration curve illustrating the dependence of ion signal on analyte mass for droplets containing 232Th. [Figure 15] FIG. 1 shows time-resolved ICP-MS peak traces for Cd-enriched Chinese hamster ovary (CHO) cell suspensions in phosphate buffered saline. [Figure 16] FIG. 1 shows time-resolved ICP-MS peak traces for a Pb-enriched aqueous suspension of Chinese Hamster Ovary (CHO) cells. [Figure 17] FIG. 2 is an expanded view of a portion of the ICP-MS system of FIG. 1, with highly schematic illustrations of the sample preparation system, flow splitter, and sample introduction system. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0060] Preferred embodiments of the invention are described below with reference to the drawings, which are intended to illustrate presently preferred embodiments of the invention and not to limit it.

[0061] 1 illustrates in highly schematic form an ICP-MS system according to an exemplary embodiment of the present invention, comprising an ICP source 100 generating an argon plasma and a mass spectrometer 200 connected to the outlet end of the ICP source.

[0062] In this example, the ICP source 100 comprises an RF generator 110 mounted on an elevator 150 so as to be adjustable at a height h relative to the mass spectrometer 200. The RF generator 110 supplies an alternating current, typically in the frequency range of 10 MHz to 100 MHz, e.g., 27 MHz or 40 Mhz, to a resonant circuit comprising a capacitor 112 and an induction coil 115. The capacitor 112 and one end of the induction coil 115 are electrically connected via a vertical metal bar 113 and a horizontal metal bar 114. The metal bar 113 and the metal bar 114 are preferably made of a highly conductive metal such as copper, silver, or gold, and are supported by a PTFE support 116. Similarly, the other end of the induction coil 115 is electrically connected to the RF generator via another horizontal metal bar 114 and another vertical metal bar 113. The horizontal metal bar 114 allows the induction coil 115 to be positioned at a sufficiently large lateral distance from the RF generator 110 that the mass spectrometer 200 can be positioned below the induction coil 115 without interfering with the RF generator 110. A typical length for the horizontal metal bar 114 is between 10 cm and 15 cm, in particular about 20 cm.

[0063] The ICP source 100 further comprises a torch box 120. The torch box 120 comprises a housing made of a conductive metal to electromagnetically protect its interior. A plasma torch 130, which will be described in more detail below in conjunction with FIG. 2, is disposed within the torch box 120 along with an induction coil 115, a horizontal metal bar 114, and a vertical metal bar 113. The induction coil 115 surrounds the lower portion of the plasma torch 130. A horizontal cooling plate 140 is disposed above the induction coil 115. The cooling plate 140 is supported within the torch box 120 by two vertical fluid-cooled metal panels 141 (cooled, for example, by water or a cooling fluid such as a mixture of ethylene glycol and water). The cooling plate 140 protects the components within the torch box 120 disposed above the induction coil 115 from convection heat generated during operation of the ICP source. A vent grid 121 allows gases and heat to escape from the torch box 120 .

[0064] At the exit end 122, the torch box 120 opens downwards and forms an extraction opening for connecting the ICP source 100 to a mass spectrometer 200. The mass spectrometer comprises a vacuum interface 210, which will be described in more detail below in conjunction with FIG. 2, followed by an ion optics unit 215, an inlet baffle 220, a quadrupole mass analyzer 230, an outlet baffle 240, and a detector 250. A mechanical pump 261 evacuates the area of ​​the vacuum interface 210. A turbomolecular pump 262 evacuates the area of ​​the ion optics unit 215, the mass analyzer 230, and the detector 250. Such types of mass spectrometer configurations are known in the art. However, whereas extraction of ions from an ion source usually occurs horizontally, the axis of ion extraction in the configuration of FIG. 1 is oriented vertically downwards.

[0065] FIG. 2 shows the plasma torch 130 and the interface 210 to the mass spectrometer 200 in more detail. The plasma torch 130 is constructed in a manner known in the art. It comprises three concentric tubes 131, 132, 133. A flow of gas (typically argon) that forms the plasma passes between the outer tube 133 and the middle tube 132 at a flow rate typically between 12 l / min and 17 l / min. A second gas flow passes between the middle tube 132 and the central tube 131 at a flow rate of 1 l / min and is used to maintain the plasma and to adjust the position of the base of the plasma relative to the ends of these tubes. The central tube 131, often called the injection tube, is used to feed the sample into the plasma.

[0066] The plasma torch 130 is centrally positioned within the induction coil 115. When RF power is applied to the induction coil 115, a strong electromagnetic field is generated inside the induction coil 115. A high voltage spark can be used to ionize a portion of the gas inside the induction coil 115, and the RF electromagnetic field accelerates the electrons and ions to sustain the plasma. The coupling between the coil and the plasma is inductive, i.e., the energy transfer is mostly due to the magnetic field generated by the oscillating current in the induction coil, and only slightly due to the electric field along the induction coil.

[0067] The sample is introduced into the plasma under the action of gravity through a central tube (injection tube) 131. The plasma is hot enough to progressively vaporize the sample, dissociate the sample, and ionize the elements contained within the sample.

[0068] The interface 210 to the mass spectrometer 200 is formed between a first baffle 211 with a small opening in a so-called sampler cone 212 and a second baffle 213 with a small opening in a so-called skimmer cone 214. A mechanical pump is connected to evacuate the space between the first baffle 211 and the second baffle 212. Ions that pass through both the sampler cone 212 and the skimmer cone 214 enter a space that is now under high vacuum and are focused into the mass analyzer by an ion optics unit 215.

[0069] FIG. 2 further shows, in a schematic way, how the ICP-MS system of the present invention can be connected to an electrostatic cell sorter 320 which is connected to a flow cytometer 310. Such a combination of a flow cytometer in cooperation with a cell sorter is known in the art as a fluorescence activated cell sorter. In the flow cytometer 310, a cell suspension 311 is fed to a nozzle 312. Droplets containing single cells are produced by the nozzle 312. Each droplet is interrogated by one or more laser beams produced by one or more lasers 313. The light along the direction of illumination and the fluorescence emitted from the droplets are detected by a light detection system 314 and analyzed by an analyzer 315. Depending on the results of the analysis, the analyzer 315 applies a charge of a given sign to the droplets before they are ejected from the nozzle by applying a positive or negative charge to the nozzle tip or a charging electrode. In the cell sorter 320, the droplets are deflected according to their charge state by a deflection plate 321 to which a DC voltage is applied. Depending on the charge state, the droplets end up in a collection vessel 322 or in a sample inlet 323. From the sample inlet 323, the droplets are transported directly downwards into the injection tube 131 of the plasma torch 130, where they fall vertically under the action of gravity.

[0070] The cell sorter 320 can be omitted and the droplets exiting the flow cytometer 310 can be introduced directly into the plasma by placing the inlet of the ICP source directly below the output nozzle of the flow cytometer. No additional nebulization system is required and 100% transport efficiency can be obtained.

[0071] For each individual droplet, whether or not the droplet has been sorted, the results from the flow cytometer 310 can be correlated with the results from the mass spectrometer 200. Combining flow cytometry with mass spectrometry in the currently proposed configuration increases the number of cellular parameters that can be simultaneously quantified when compared to flow cytometry alone or mass spectrometry alone. To that end, the analyzer 315 can be configured to perform the correlation between the results of flow cytometry and mass spectrometry, or a separate correlation can be provided.

[0072] 3 shows in more detail the cold plate 140 and the vertical metal panel 141. The cold plate 140 has a cutout 142 that accommodates the plasma torch when the cold plate is installed. The vertical panel is provided with a number of channels or perforations 143 that extend into each face of the panel for the passage of a cooling fluid, such as cold air or water, to actively cool the cold plate.

[0073] FIG. 4 shows a variant in which the plasma torch 130' is configured as a so-called direct injection high efficiency atomizer (DIHEN), which allows direct sample introduction into the plasma without the need for a separate aerosolization device. A high efficiency atomizer 134 replaces the injection tube in the plasma torch 130'. This allows the sample to be directly introduced into the plasma without the need for prior aerosolization. This results in a transport efficiency of 100%. Such direct injection atomizers are known per se (A. Montaser, "Inductively Coupled Plasma Mass Spectrometry", Wiley-VCH: Washington DC, 1998). However, whereas they are usually operated in a horizontal configuration, in the present invention the direct injection atomizer operates in a vertical configuration.

[0074] Figures 5-8 show four examples of aerosolization devices that are connected to the plasma torch 130 of the ICP source shown in Figure 1. These figures are schematic and the relative dimensions of the components shown in these figures are not to scale.

[0075] In FIG. 5, a droplet delivery and low-temperature desolvation system 410 is shown. The droplet dispenser 411 is connected to a flow adapter 412 mounted on the top of a so-called drop tube 413. The drop tube 413 is a 40 cm long stainless steel tube with an inner diameter of 4 mm. A mixture of argon and helium is used for desolvation of the droplets inside the drop tube 413. Desolvation of the droplets leads to a reduction in local cooling of the plasma. For further details, see "[2]". Whereas in the prior art, the aerosolized sample leaving the drop tube 413 is transported horizontally to the ICP source using a carrier gas flow, in this example the aerosolized sample is introduced directly downwards into the injection tube 131 of the plasma torch without the need for an additional carrier gas flow. This ensures that all sample droplets or particles leaving the drop tube 413 reach the plasma, regardless of their size and regardless of the gas flow rate.

[0076] In Fig. 6, aerosolization is performed by a so-called total consumption sample introduction system 420. A low-flow nebulizer 421, which typically operates at a liquid flow rate of less than 10 μL / min, is connected to a small-volume nebulizer chamber 422. The nebulizer chamber 422 pushes the droplets generated by the nebulizer 421 downward along the central axis and prevents the droplets from adhering to the walls of the nebulizer chamber 422. The nebulizer chamber 422 is open at the bottom and forms an outlet. The outlet is directly connected to the injection tube 131 of the plasma torch 130.

[0077] 7, a solid sample 432 is aerosolized in a laser ablation cell 430 using a laser 431 that directs an intense pulsed laser beam at the back side of the sample 432, facing away from the ICP source. The induced shock wave causes ejection of the sample material, which falls into the injection tube 131 of the plasma torch 130 ("back side ablation").

[0078] 8 shows aerosolization using a laser ablation cell 440 according to U.S. Pat. No. 9,922,811. A laser 441 irradiates the surface of a solid sample 442 with laser light, causing the sample material to break into smaller pieces. The smaller pieces enter a channel 443, which transports the material directly down into the inlet tube 131 of the plasma torch 130. For further details regarding the laser ablation cell 440 and its mode of operation, see U.S. Pat. No. 9,922,811. Unlike the prior art where the channel 443 is oriented horizontally and the laser beam strikes the sample surface vertically, in this example the channel 443 is oriented vertically and the laser beam is oriented horizontally.

[0079] The presently proposed ICP source can be used with any type and configuration of mass analyzer. For example, FIG. 9 shows a TOF mass analyzer 200' that can be used in conjunction with the ICP source shown in FIGS. 1 and 2. The interface 210 is configured similarly to FIGS. 1 and 2. Ions extracted from the ICP source through the interface 210 pass through a notch filter 260 and an extractor 270 into a TOF mass analyzer 280. Any other type and configuration of mass analyzer can be employed. For example, a quadrupole mass analyzer, a sector field mass analyzer, or an ion trap mass analyzer can be used instead. If necessary, electrostatic mirrors and / or deflection magnets can be used to deflect the ions into the mass analyzer.

[0080] FIG. 10 shows a Microwave Inductively Coupled Atmospheric Pressure Plasma (MICAP) source 500 configured for vertical sample introduction. The MICAP source comprises an RF generator (microwave generator) 510 in the form of a magnetron. A fan 511 provides cooling for the magnetron 510. An antenna 512 couples the microwave field to the interior of a torch box 520. An annular dielectric resonator 515 is placed within the microwave field. The resonator exhibits a bulk polarization current around the ring, which oscillates at the microwave frequency. A plasma torch 530 is placed above the resonator 515. The plasma torch 530 is configured in the same manner as the plasma torch 130 described in conjunction with FIG. 2. Nitrogen gas passes into the central region of the resonator 530. Nitrogen plasma is sustained by an electromagnetic field at the center of the resonator 515 by inductive coupling. For more details, see US Pat. No. 5,399,413 and US Pat. No. 5,433,636. In the prior art, the plasma torch of a MICAP source is oriented horizontally, whereas in this example, the plasma torch 530 is oriented vertically and the sample is introduced through a central tube 531 in the direction of the gravity vector.

[0081] 11 to 16 show experimental results obtained with the configuration shown in FIG.

[0082] FIG. 11 and FIG. 12 are 133 Cs (Figure 11) and 232 The droplet generator was used to generate a time-resolved ICP-MS peak trace of monodisperse droplets containing 20 ppb of Th (Figure 12). 133 Cs and 232 Droplets with a diameter of 66 μm were generated from a solution containing both Th and Zn at 20 Hz. All ejected droplets reached the plasma and were successfully detected by ICP-MS. 133 Cs has an ionization energy of 3.89 eV, 232 Th has an ionization energy of 6.31 eV. These results demonstrate that the ICP-MS system of the present invention can easily achieve 100% transport efficiency.

[0083] FIG. 13 and FIG. 14 are 133 Cs (Figure 11) and232 12 shows the dependence of ion signal on analyte mass for droplets containing Th (FIG. 12). These results are 133 Cs and 232 Both figures were obtained by investigating monodisperse droplets with different analyte concentrations of 2 ppb (left-most data point), 5 ppb, 10 ppb, and 20 ppb (right-most data point) for both Th. Both figures show a linear relationship between analyte mass and ion signal, indicating complete desolvation, evaporation, dissociation, and ionization of the droplets using the vertical plasma configuration of the present invention.

[0084] Figure 15 shows time resolved ICP-MS peak traces for a Cd-enriched suspension of Chinese Hamster Ovary (CHO) cells in phosphate buffered saline. Figure 16 shows time resolved ICP-MS peak traces for a Pb-enriched suspension of Chinese Hamster Ovary (CHO) cells in water. These traces show that cells can be successfully introduced into the plasma using the vertical configuration of the present invention. Since 100% transport efficiency is expected, the "missing" peaks are likely due to droplets that do not contain cells. On the other hand, it cannot be excluded that other droplets contain multiple cells.

[0085] 17 shows a schematic diagram of the connection of an ICP-MS system according to an embodiment of the present invention to a continuous sample delivery system with a sample pre-treatment device 610. The sample pre-treatment device 610 can be a separation device, for example a chromatography device such as an HPLC or GC device, or an electrophoresis device such as a CE device. In other embodiments, the sample pre-treatment device can be any other device that performs some analysis or treatment on an incoming fluid stream containing the sample.

[0086] The sample pretreatment device 610 has an inlet 611 and an outlet 612. The sample pretreatment device receives a continuous inlet fluid stream F at its inlet 611. in The sample preparation device 610 receives at its outlet 612 a continuous output fluid stream F out If the sample pretreatment device 610 is a separation device, the output fluid stream Fout can have a time-varying composition, and the separation device can be configured to determine information about this composition, as is known in the art.

[0087] The flow splitter 620 is connected to the output of the sample preparation device 610. The flow splitter 620 provides a continuous output fluid flow F out Let F be the sample flow. sample and residual fluid flow F res The sample flow is divided into two. sample The flow rate of the residual fluid F res Except for the different flow rates, the composition of the two fluid streams may be identical.

[0088] Sample flow F sample enters the sample introduction system, which in this example is configured as a droplet delivery and desolvation system as described above in conjunction with Figure 5. However, any other sample introduction system that allows a small continuous stream of the fluid sample to be introduced into the plasma of the ICP source can also be employed. In particular, the sample introduction system can be a total consumption system as in Figure 6 or can comprise a DIHEN nebulizer as in Figure 4. The sample introduction system can be integrated with the ICP source to form a single unit.

[0089] Some sample pretreatment devices, particularly separation devices such as HPLC devices, generally have a flow rate at the outlet that is significantly higher than the flow rate required for the sample introduction system (e.g., 0.1 μl / min to 10 μl / min using a droplet generator at 1000 Hz), and thus a flow splitter 620 is provided. In some other sample pretreatment devices, such as some CE devices, the flow splitter 620 may not be necessary and can be omitted.

[0090] The sample introduction system is positioned above the ICP source such that the sample is introduced into the ICP source in a vertically downward direction G (ie, in the direction of gravity).

[0091] The correlator 630 receives separation information for the various fractions exiting the sample preparation device 610. Additionally, the correlator 630 receives mass information for the mass distribution in the fractions from a mass spectrometer connected to the ICP source. The correlator 630 correlates the separation information with the mass information, for example in the form of LC / MS information, GC / MS information, or CE / MS information, as is known in the art.

[0092] Many modifications are possible without departing from the scope of the invention. In particular, other types of ICP sources than the two types mentioned above can be employed. The sample can be divided into small droplets or particles in other ways than those mentioned above. Any type and orientation of mass spectrometer can be employed. [Explanation of symbols]

[0093] 100 ICP sources 110 RF generator 112 Plate Capacitor 113 Vertical Metal Rod 114 Horizontal metal bar 115 Induction Coil 116 PTFE support 120 Torch Box 121 Exhaust grid 122 Outlet end 130, 130' Plasma Torch 131 Central pipe (injection pipe) 132 Middle tube 133 Outer tube 134 Sprayer 140 Cooling plate 141 Vertical Panel 142 Notch 143 Perforation 150 Elevator 200 Quadrupole mass spectrometer 200' TOF mass spectrometer 210 Interface 211 First Baffle 212 Sampler Cone 213 Second Baffle 214 Skimmer Cone 215 Ion Optical Unit 220 Inlet baffle 230 Quadrupole mass spectrometer 240 Exit baffle 250 detector 260 Notch Filter 270 Extractor 280 TOF mass spectrometer 261 Mechanical Pump 262 Turbomolecular Pump 310 Flow Cytometer 311 Slurry 312 Nozzle 313 Laser 314 Detection Systems 315 Analyzer 320 Cell Sorter 321 Deflection plate 322 Collection Container 323 Sample inlet 410 Droplet Delivery and Desolvation System 411 Droplet Dispenser 412 Flow Adapter 413 Drop tube 420 Total consumption type sample introduction system 421 Sprayer 422 Spray room 430 Laser Ablation Cell 431 Laser 432 Samples 440 Laser Ablation Cell 441 Laser 442 Samples 443 Flow Path 500 MICAP sources 510 RF generator (magnetic tube) 511 Fan 512 Antenna 515 resonator 520 Torch Box 530 Plasma Torch 531 Injection tube 610 Sample preparation equipment 611 Entrance 612 Exit 620 Diversion device h Height G Gravity direction F in inflow flow F out Outflow F sample Sample Flow F res Residual flow

Claims

1. 1. An ionization system comprising an ICP source for generating ions from a sample using inductively coupled plasma, the ICP source (100; 500) configured to be connected to a mass spectrometer (200; 200'), the ICP source (100; 500) configured to introduce the sample vertically downwards (G) into the plasma, A flow cytometer (310) and / or a cell sorter (320), The ICP source (100; 500) is connected to the flow cytometer (310) or the cell sorter (320) such that the sample is transferred in the vertical downward direction (G) from the flow cytometer (310) or the cell sorter (320) to the ICP source (100; 500).

2. 2. The ionization system of claim 1, further comprising a correlator (315) configured to receive flow cytometry information from the flow cytometer (310) and mass spectrometry information from a mass spectrometer (200; 200') connected to the ICP source (100; 500), and to correlate the flow cytometry information with the mass spectrometry information to obtain combined flow cytometry and mass spectrometry information for individual cells or particles.

3. 1. An ionization system comprising an ICP source for generating ions from a sample using inductively coupled plasma, the ICP source (100; 500) configured to be connected to a mass spectrometer (200; 200'), the ICP source (100; 500) configured to introduce the sample vertically downwards (G) into the plasma, A continuous flow sample delivery system (610, 620); A sample introduction system (410), The sample introduction system (410) provides a continuous sample stream (F) containing the sample without the need for intermediate offline storage. ｓａｍｐｌｅ ) is connected to the continuous flow sample delivery system (610, 620) such that the sample is transferred from the continuous flow sample delivery system (610, 620) to the sample introduction system (410); The sample introduction system (410) is configured to introduce the sample into the ICP source (100; 500) in the vertically downward direction (G) without the need for intermediate offline storage.

4. The continuous flow sample delivery system (610, 620) comprises a continuous flow sample preparation device (610), which supplies a continuous inlet flow (F ｉｎ ) and a continuous output flow (F ｏｕｔ and an outlet configured to provide the continuous outlet flow (F ｏｕｔ At least a portion of the continuous sample flow (F ｓａｍｐｌｅ ), and the continuous flow sample preparation device (610) comprises the continuous inlet flow (F ｉｎ 4. The ionization system of claim 3, configured to perform at least one of the following actions on the ionizing gas: analyzing and / or modifying the composition thereof.

5. The continuous flow sample preparation device (610) is a ｉｎ ) and separating the chemical species or particulate matter in said continuous effluent stream (F ｏｕｔ 5. The ionization system of claim 4, comprising a separator configured to elute into the ionizer.

6. The ionization system of claim 4 or 5, further comprising a correlator (630) configured to receive analytical information from the continuous flow sample preparation device (610) and mass spectrometry information from a mass spectrometer (200; 200') connected to the ICP source (100; 500), and to correlate the analytical information with the mass spectrometry information.

7. The continuous flow sample delivery system (610, 620) provides the continuous output flow (F ｏｕｔ ) into a continuous sample flow (F ｓａｍｐｌｅ ) and residual flow (F ｒｅｓ The ionization system according to any one of claims 4 to 6, further comprising a flow dividing device (620) for dividing the flow continuously into the first and second ions.

8. The ionization system of any one of claims 1 to 7, wherein the ICP source is configured such that ions are extracted from the plasma in the vertically downward direction (G).

9. 9. The ionization system of claim 1, comprising a plasma torch comprising an injection tube defining a vertically oriented longitudinal axis, the injection tube having an upper end configured to receive the sample and a lower end configured to introduce the sample into the plasma.

10. 10. The ionization system of claim 1, comprising an electromagnetic coupling element for sustaining the plasma by inductive coupling, the electromagnetic coupling element defining a longitudinal axis, the longitudinal axis being oriented vertically.

11. 1. An ICP-MS system comprising: An ionization system according to any one of claims 1 to 10; a mass spectrometer (200; 200') connected to the ICP source (100; 500) to receive ions from the ICP source (100; 500); An ICP-MS system comprising:

12. 1. A method of mass spectrometry comprising: maintaining a plasma using an ICP source (100; 500); introducing a sample into the plasma and generating ions from the sample; Analysing the mass-to-charge spectrum of ions extracted from the plasma using a mass spectrometer (200; 200') connected to the ICP source (100; 500); Including, The sample is introduced into the plasma in a vertically downward direction (G); The method comprises: Analyzing at least one characteristic of the sample using a flow cytometer (310) and / or sorting sample droplets according to at least one characteristic using a cell sorter (320); transferring the sample vertically downward (G) from the flow cytometer (310) or the cell sorter (320) into the ICP source (100; 500); The method of claim 1, further comprising:

13. 1. A method of mass spectrometry comprising: maintaining a plasma using an ICP source (100; 500); introducing a sample into the plasma and generating ions from the sample; Analysing the mass-to-charge spectrum of ions extracted from the plasma using a mass spectrometer (200; 200') connected to the ICP source (100; 500); Including, The sample is introduced into the plasma in a vertically downward direction (G); The method comprises: A continuous sample stream (F) containing the sample is produced without the need for intermediate offline storage. ｓａｍｐｌｅ ) from a continuous flow sample delivery system (610, 620) to the sample introduction system (410); The method of claim 1, wherein the sample is introduced into the plasma in the vertically downward direction (G) by the sample introduction system (410) without the need for intermediate offline storage.

14. The continuous flow sample delivery system (610, 620) comprises a continuous flow sample preparation device (610) having an inlet (611) and an outlet (612); The method comprises: A continuous inlet flow (F) containing the sample is provided at the inlet (611). ｉｎ ) and The continuous inlet flow (F ｉｎ ) performing at least one of analyzing and / or modifying its composition; A continuous output stream (F) containing the sample is provided at the outlet (612). ｏｕｔ ) and The continuous outflow (F ｏｕｔ ) into at least a portion of the continuous sample flow (F ｓａｍｐｌｅ ) and The method of claim 13, comprising:

15. The continuous flow sample preparation device (610) comprises a separation device; The method comprises: The separation device is used to separate the continuous inlet stream (F ｉｎ ) and separating the chemical species or particulate matter in said continuous effluent stream (F ｏｕｔ 15. The method of claim 14, comprising eluting into

16. The continuous outflow (F ｏｕｔ ) into the continuous sample flow (F ｓａｍｐｌｅ ) and residual flow (F ｒｅｓ 16. The method of claim 14 or 15, further comprising continuously splitting the flow into

Images