Laboratory sample analyzer, liquid chromatography cartridge, and method for heating a sample

Abstract

According to the present invention, there is provided an analyzer comprising: an interface for receiving a cartridge (3) comprising one or more liquid chromatography columns (4, 5) for performing an analytical task and connecting it to the analyzer; a heater (11) for heating the chromatography columns (5, 6); the cartridge (3) comprising a cartridge casing (6) for accommodating the liquid chromatography columns (4, 5); a heating block (7) thermally connected to the liquid chromatography columns (5); and a thermal interface (8) for thermally connecting the heating block (7) to the heater (11).

Description

[Technical field]

[0001] The present invention relates to the preparation and analysis of laboratory samples on an analyzer utilizing replaceable liquid chromatography cartridges. More specifically, the present invention relates to liquid chromatography columns and the heating of samples therein. [Background technology]

[0002] Liquid chromatography (LC) is well known in the fields of chemical separation, compound purification, and chemical analysis. The central component of a liquid chromatography system is the chromatography column. The column comprises a tube, e.g., a capillary tube, packed with a permeable solid material that is itself a chromatographic stationary phase or that contains or supports a chromatographic stationary phase. A fluid mixture containing both the compound of interest for purification or separation, as the chromatographic mobile phase, is flowed under pressure through the column from the inlet to the outlet end.

[0003] One or more liquid chromatography columns may be placed in a replaceable cartridge that can be changed at the end of its useful life or when the type of column needs to be changed. One such system is described in US2017 / 0023536. Another document disclosing a chromatography cartridge is US Patent Application Serial No. 13882116.

[0004] Further documentation relating to liquid chromatography and related systems and analyzers is disclosed in the documents set forth in the citation list at the end of this specification. Summary of the Invention

[0005] The invention is defined by the features of the independent claims. Some particular embodiments are defined in the dependent claims.

[0006] According to a first aspect of the present invention, there is provided an analyzer comprising: an interface for receiving a cartridge comprising one or more liquid chromatography columns and connecting it to the analyzer to perform an analytical task; a heater for heating the chromatography columns; a cartridge comprising a cartridge casing for housing the liquid chromatography columns; a heating block thermally connected to the liquid chromatography columns; and a thermal interface for thermally connecting the heating block to the heater.

[0007] According to a second aspect of the present invention, there is provided a liquid chromatography cartridge comprising a cartridge casing for housing a liquid chromatography column, a heating block thermally connected to the liquid chromatography column, and a thermal interface for thermally connecting the heating block to a heater.

[0008] According to a third aspect of the invention there is provided an analyser, wherein the heater comprises a thermistor for measuring temperature, the thermistor being thermally connectable to a thermal interface of the cartridge.

[0009] According to a fourth aspect of the invention there is provided an analyser, wherein the heater comprises a heat transfer part movably mounted to the heater to provide alignment with a thermal interface of the cartridge.

[0010] According to a fifth aspect of the invention there is provided an analyser, wherein the heater comprises a pressing element for pressing the heat transfer part against the thermal interface.

[0011] According to a sixth aspect of the present invention there is provided an analyser, wherein the heater comprises a thermistor for measuring temperature and a resistor for generating heat, the thermistor and the resistor being thermally connectable to a thermal interface of the cartridge. [Brief description of the drawings]

[0012] [Figure 1] 1 illustrates an analyzer in accordance with at least some embodiments of the present invention. [Diagram 2] The analyzer of FIG. 1 is illustrated from another perspective. [Diagram 3] FIG. 3 is an enlarged view of the device of FIGS. 1 and 2. [Figure 4] 1 illustrates an exemplary apparatus capable of implementing at least some embodiments of the present invention. [Diagram 5] 2 is a cross-sectional view of FIG. 1 illustrating an exemplary apparatus in accordance with at least some embodiments of the present invention. [Figure 6] 6 illustrates the analyzer of FIG. 5 showing the heater in an open position. [Figure 7] 6 illustrates the heater portion of the analyzer of FIG. 5 with the heater shown in an open position and shown in partial cross-section. [Figure 8] 8 illustrates some portions of the heater section of the analyzer of FIGS.

[0013] Embodiment definition In the present context, the term "liquid chromatography" refers to the process of selectively retaining one or more components of a fluid solution as the fluid permeates uniformly through a column of fine material, or a capillary passage.

[0014] Some LC methods are very sensitive to temperature. As an example, how temperature sensitivity is currently addressed is to use a pre-heater sleeve to heat the liquid before exposing it to the analytical column. The temperature of the liquid in the column is too sensitive to the environmental temperature and the environmental airflow must be controlled to ensure it is sufficiently stable. More stable and controllable heating and temperature control can be achieved by heating the analytical column via a thermal connection to a heating block. An exemplary heating block has a thermal interface to a heater component contained in the analytical instrument. The column and heating block are housed in a cartridge casing. The cartridge is replaceable and can be installed and removed from the analyzer. The analyzer includes a heater that can be thermally connected and disconnected from the cartridge.

[0015] FIG. 1 illustrates some portions of an analyzer that can support at least some embodiments of the present invention. The analyzer includes equipment for handling and processing analytes, reagents, and samples, performing analyses, and other elements required for its operation. A general operational description of such an analyzer is disclosed below. FIG. 1 specifically illustrates an analyzer that includes an interface for receiving a cartridge that includes one or more liquid chromatography columns.

[0016] The analyzer comprises a frame 1, which includes an interface 2 for receiving a cartridge 3. The interface may comprise electrical, data and fluid connections as well as mechanical connectors for removably coupling the cartridge to the analyzer. The cartridge comprises a casing 6. The casing 6 houses a first column 4 for liquid chromatography and a second column 5 for liquid chromatography. The second column 5 is partially enclosed in a heating block 7. The heating block 7 is made of a material with good thermal conductivity capabilities and is in contact with the surface of the second column to ensure good heat transfer between the contact surfaces and good heat transfer capacity. The heating block 7 has a surface that extends outside the casing 6. This surface forms a thermal interface 8 for connecting the cartridge to a heater of the analyzer.

[0017] 2 illustrates the heater in an open position, where the thermal interface 8 can be seen exposed from the casing 6 of the cartridge 3, and the heater 11 is visible on the heater 11 facing upwards, with the heater 11 having an outwardly extending surface for connection to the thermal interface 8. The thermal interface 8 is covered with thermal tape 14.

[0018] The heater 11 is rotatably mounted on an axis 9. The heater 11 can be operated by rotating a handle 10 on the axis so that the heater 11 is lifted from the cartridge for removal of the cartridge and pressed against it for connecting the heater 11 to the cartridge 3. Another version of the heater can have a handle integrated with the casing of the electronic components, whereby rotation of the heater can be accomplished.

[0019] The heat transfer interfaces between the elements of the device, from the resistor 15 to the second column 5, are depicted in FIG. 3. The first interface is between the resistor 15 and the heat transfer part 13 of the heater 11. The resistor 15 is used to heat the heat transfer part 13 of the heater 11 and can be permanently and securely attached to the heat transfer part to ensure good heat transfer. In the device of FIG. 3, the resistor 15 is attached to the heat transfer part 13 by a screw to allow replacement of the resistor 15. The second heat transfer interface is formed between the heat transfer part 13 and the heat tape 14, and the third heat transfer interface is formed between the heating block 7 and the heat tape 14. At these interfaces the heat tape 14 is used to ensure good heat transfer. The fourth interface is between the heating block 7 and the second column 5. All other interfaces are configured to be fixed when the device is in operation and are only removed when repair or maintenance is required. One of the interfaces between the heater 11 (heat block), the thermal tape 14 and the heating block 7 is configured to be openable to allow replacement of the cartridge 3 .

[0020] Figures 6 and 7 show the heater 11 in an open position, where the cartridge can be easily replaced, and it can be seen from Figures 5 and 6 that the heater movement by rotation ensures that the heater 11 is accurately and securely positioned over the cartridge 3.

[0021] Figure 8 shows the heater 11 without the lid or cover. In this view, the spring 12 can be seen to press the heat transfer section 13 against the thermal interface 8. A resistor 15 and a thermistor 16 are also visible on the heat transfer section 13. There is also a "thermal tape" 14 between the heating block and the heat transfer section. This makes the heater element connection (or heat transfer interface) more robust against dirt, foreign objects (e.g. even a small piece of paper can cause a large loss of heat transfer capability), heater element level errors, and also prevents metal-to-metal wear.

[0022] The thermistor 16 is for measuring the temperature and is thermally connected to the thermal interface of the cartridge. Furthermore, the heater 11 of Fig. 8 comprises a heat transfer part, which is movably mounted on the heater 11 to provide alignment with the thermal interface of the cartridge. The heater 11 also comprises a pressing element for pressing the heat transfer part against the thermal interface. A resistor 15 is provided for generating heat, the thermistor and the resistor being thermally connectable to the thermal interface of the cartridge.

[0023] The cartridge may include, for example, a Quick Connect Cartridge H (Thermo Fisher Scientific). The temperature of the second column (or the first column, if desired) of the cartridge is changed and controlled with a heater called a Column Heater. The Column Heater and the Quick Connect Cartridge H are components that effectively transfer heat to the liquid chromatography column and are thermally connected. Accurate connection of these heat transfer parts is essential for accurate temperature measurement and heat transfer. -The Column Heater has a resistor that heats the column of the Cartridge through a heat transfer section. -The column heater has a thermistor for measuring temperature and is connected to the heat transfer section. -The heat transfer parts have some freedom of movement to ensure accurate positioning when the heat transfer parts are locked together. The heat transfer parts have a spring 12, which ensures a constant force between the heat transfer parts.

[0024] Users can easily remove the Column Heater from the Quick Connect Cartridge H to replace the cartridge. The position of the Column Heater is measured with a sensor. Thermal tape between the moving heat transfer parts reduces the risk of an improper heat transfer connection.

[0025] The heater 11 is not in the cartridge housing, but is part of the analyzer. The problem with cartridges is that they are consumables and may be replaced up to once a week, which makes them more costly than external heaters, makes cartridges with heaters more difficult to recycle, and makes quality control of large volume assemblies more difficult. In cartridges according to at least some embodiments of the invention, only the simplest mechanical parts are located in the cartridge, while electronic components and moving mechanical parts are located on the external heater. The heater elements and parts can be easily (user friendly) removed and reinstalled to the column through the cartridge for reliable (counter design features against user error) and accurate (counter design features against wear and tear of the mechanism) heat transfer. At least some embodiments of the invention answer the problem of how to mechanically make a removable external heater that can make a sufficiently good contact through the heat transfer part.

[0026] In the column heater mechanism described above, the column is located in a cartridge casing (described in US2017 / 0023536A1 and its parent application, U.S. Patent Application No. 13 / 882,116, filed as Application No. PCT / US11 / 58229), heat is transferred to the column surface, and temperature is measured from the column surface via contacting heat transfer parts, not air. These mechanical parts come into contact when the cartridge is inserted (Clamp Heater concept) or when the column heater casing (which houses the heat transfer parts, temperature measurement sensor, and heat generating components) is moved to its locked position. The primary direction of movement is rotational or linear, not sliding or lateral, and brings two opposing flat surfaces into direct flush alignment. Also, there are either: - moving the heater casing (containing the heating and sensing components) into contact with the stationary column portion; or - The column portion comes into contact with the fixed heating and sensing components during installation of the cartridge by the user. Possibility to use a locking handle to secure the position of the cartridge or the heating / sensor part.

[0027] Some known solutions have the problem that heat is transferred to the column and the temperature is measured through the air. This makes the temperature transfer slow and the measurement sensitive to environmental influences (such as outside temperature and air movement) and inaccurate. It is important to have good mechanical contact during the heat transfer to provide fast heat transfer and fast and accurate temperature measurement. These challenges can be addressed by at least some embodiments of the present invention.

[0028] Also, some known solutions do not account for washing the column compartment between column exchanges and during rotational movement, thus posing a risk of cross contamination, and if the ends of the columns are not individually sealed with their respective seals when the columns are not in use, there is a risk of contamination. The cartridges (with columns therein) of the present disclosure are easily exchanged and each have a sealing cap when not in use.

[0029] FIG. 4 illustrates one example of an apparatus capable of implementing at least some embodiments of the present invention.

[0030] The heat transfer portion 13 of the heater 11 is spring loaded on all four corners for even leveling and thermal contact pressure against the heater block in the cartridge 3. This feature is for robustness against manufacturing tolerances and wear that may occur with use. Thermal tape 14 between the heat transfer portion 13 of the heater 11 and the heating block 7 provides enhanced robustness against thermal contact. This feature is for robustness against manufacturing tolerances and small particles that may occur between the heater elements.

[0031] The column temperature can be precisely controlled (to ±2° C.). The heater element is located outside the Quick Connect Cartridge but is rigidly connected to a heater block that surrounds the column itself. This allows the heater to control the required temperature in the column as required by the assay while allowing easy replacement of the Quick Connect Cartridge.

[0032] The heater block movement can be done in several ways or mechanisms. The described example provides the correct functionality (heat transfer and temperature measurement). It is important that the connection of the column to the heater block, the heater block to the heat transfer part and the heat transfer part to the thermistor are tight and always the same heat transfer cross section at the connection, but it is also relevant that the temperature measurement is done exactly near the column and the same every time the cartridge is connected in order to control the heat provided.

[0033] In the following, some aspects of sample processing and analysis methods related to liquid chromatography are described.

[0034] The analyzer can provide a fully automated random access workflow for the analysis of clinical patient samples. WO2012 / 058632 discloses an automated system for sample preparation and analysis that is particularly suitable for use in connection with at least some embodiments of the present invention. The system incorporates the following features online: sample preparation, TurboFlow and high performance liquid chromatography (HPLC) technologies, multiplexing technology, and tandem mass spectrometry detection in selected reaction monitoring (SRM) mode. The multiplexing technology is a configuration that combines two LC channels that can run simultaneously into one mass spectrometer, maximizing sample throughput. SRM is a highly selective technique used to detect specific analytes using tandem mass spectrometry.

[0035] Patient samples are introduced to the analyzer by placing sample tubes or cups in a sample rack and inserting the sample rack into the analyzer. Similarly, reagents are introduced to the analyzer by placing them in a reagent rack and inserting the reagent rack into the analyzer. Samples and reagents in the rack are automatically identified from their respective barcodes. If barcodes are not used, samples can be identified manually using the analyzer software. Samples are defined via the LIS or locally using the analyzer software. The analyzer accepts most common types of sample tubes and barcodes. Sample material can be, for example, whole blood, plasma, or serum.

[0036] Biological samples contain various endogenous matrix compounds, such as proteins and lipids. These low volatility compounds can interfere with the delicate ion formation process in the ion source of the mass spectrometer, i.e., cause ion suppression or ion enhancement. Therefore, proper pretreatment is necessary before introducing biological samples into the LC / MS system. During the sample preparation process, the amount of interfering compounds from the sample matrix is ​​significantly reduced, which in turn enhances the detection sensitivity and improves the instrument performance by reducing the expected LC column blocking and MS contamination.

[0037] Although many of the proteins present in a given matrix may precipitate out of solution during sample preparation, many components still remain in solution that may cause interference during analysis. These are removed using TurboFlow technology, an on-line clean-up technique used to separate analytes from interfering compounds. TurboFlow technology provides rapid and efficient separation of complex sample matrices and analytes.

[0038] In TurboFlow technology, the sample is introduced into a TurboFlow column with a high velocity solvent flow. The column is packed with large column particles that interact with small molecules, while large molecules, such as proteins, flow through the column and are discarded. Separation of molecules is based on the different diffusion properties between large and small molecules and the chemical binding affinity of the diffused small molecules to the column particles.

[0039] High performance liquid chromatography (HPLC) is a chemical and physical separation used to separate a mixture in solution into its individual components. The analyzer utilizes partition chromatography, where the separation is based on the components of the sample distributing between a mobile phase and a stationary phase. The mobile phase is made up of various ratios of solvents, and the stationary phase is the solid packing of the analytical column in the quick-connect cartridge.

[0040] Components with a composition ratio that favors the stationary phase will interact more, or more strongly, with the active groups of the column packing and therefore will require longer to pass through the LC channel. To elute the analytes and other compounds bound to the stationary phase, the solvent composition of the mobile phase must be changed so that the analytes separate from the stationary phase into the mobile phase by a change in polarity. The time it takes for an analyte to pass through the analytical column is called the retention time. Retention time depends on the analyte, the strength of the interaction with the stationary phase, the composition of the solvent used, the temperature, and the flow rate of the mobile phase.

[0041] Analytes are first extracted from the sample matrix in an automated sample preparation process. The injected sample extract (supernatant) is then purified with TurboFlow technology and the analytes are chromatographically separated from interfering compounds with HPLC. Selected analytes are then detected and measured with a mass spectrometer. The results are quantified and analyte concentration values ​​are displayed to the user.

[0042] The user inserts a sample rack with a homogenous, thoroughly mixed sample and a reagent rack with the required reagents into the analyzer. Whole blood samples in tubes are automatically mixed on-board using mixing tips before dispensing. The extraction vessel is brought to the vessel disc and opened for dispensing. Reagents including internal standards and samples are dispensed using assay specific parameters.

[0043] The internal standard solution in an organic solvent causes protein precipitation. In the case of a whole blood sample, the reagent ruptures the red blood cells in the sample releasing the analyte, which then precipitates the protein. The contents of the extraction vessel are mixed on a shaker. Shaking enhances protein precipitation and lysis of the whole blood. The sample and reagent are mixed to form a homogenous mixture / solution. The supernatant is injected into the injection loop. The injector loop is brought online and the sample is mixed with the LC solvent stream. The sample stream enters the TurboFlow column. Small molecules such as the analyte and internal standard are separated from larger molecules such as proteins. Large (interfering) molecules are washed away to waste, and then the analyte and internal standard are flushed back from the TurboFlow column to the analytical column. The analyte and internal standard concentrate in sharp bands as they enter the analytical column. The analyte and internal standard are separated from other compounds in the sample according to their respective polarities and affinities for the stationary phase. By increasing the organic solvent flow rate, the analytes and internal standards are released and eluted from the analytical column into the mass spectrometer.

[0044] An example of a cartridge exchange is as follows.

[0045] The cartridge 3 can be accessed by opening the LC door. The cartridge 3 can be released by opening the handle 10 of the quick connect clamp, releasing the quick connect cartridge. The column heater 11 can be opened by pressing the lock button on the handle 10 and pulling the handle. A dialogue box opens indicating the quick connect clamp is open. The old quick connect cartridge 3 can be extracted from the clamp. The used cartridge 3 can be discarded or stored for later use. A new cartridge 3 can now be inserted into the clamp by inserting the USB connector of the cartridge 3 into the USB port of the clamp and pressing the cartridge 3 into place. Close the column heater handle 10 by pressing the handle 10 towards the analyzer until a click is heard. The handle and door of the quick connect clamp are closed.

[0046] It is to be understood that the disclosed embodiments of the invention are not limited to the particular structures, process steps, or materials disclosed herein, but extend to equivalents thereof as recognized by those skilled in the art. It is also to be understood that the terminology used herein is used only for the purpose of describing particular embodiments, and is not intended to be limiting.

[0047] References throughout this specification to "one embodiment" or "an embodiment" mean that a particular feature, structure, or characteristic described in connection with that embodiment is included in at least one embodiment of the invention. Thus, appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment.

[0048] As used herein, a plurality of items, structural elements, compositional elements, and / or materials may be presented in a common list for convenience. However, these lists should be construed as if each member of the list were individually identified as a separate and unique member. Thus, the individual members of such lists should not be construed as de facto equivalents to other members of the same list solely based on presentation in a common grouping, absent indication to the contrary. Furthermore, various embodiments and examples of the invention may be referenced herein along with alternatives for the various components thereof. It is understood that such embodiments, examples, and alternatives should not be construed as de facto equivalents to one another, but should be considered as separate and autonomous manifestations of the invention.

[0049] Furthermore, the described features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. In the following description, numerous specific details are provided, such as examples of lengths, widths, shapes, etc., to provide a thorough understanding of embodiments of the invention. However, one of ordinary skill in the relevant art will recognize that the invention can be practiced without one or more of the specific details, or with other methods, components, materials, etc. In other instances, well-known structures, materials, or operations are not shown or described in detail to avoid obscuring aspects of the invention.

[0050] While the foregoing examples illustrate the principles of the present invention in one or more particular applications, it will be apparent to those skilled in the art that numerous changes in form, use, and details of implementation may be made therein without the exercise of inventive faculty and without departing from the principles and concepts of the invention. Accordingly, it is not intended that the present invention be limited, except as by the scope of the claims which follow.

[0051] The verbs "comprise" and "include" are used in this specification as open limitations which neither exclude nor require the presence of unrecited features. Features recited in the dependent claims are mutually freely combined unless expressly stated otherwise. Furthermore, it is to be understood that the use of "a" or "an", i.e. the singular, throughout this specification does not exclude the plural.

[0052] Industrial Applicability The present invention can be utilized in chemical analyzers and in the manufacture of analyzers. [Table 1] [Table 2]

[0053] Citation List Patent Literature US2017 / 0023536, US13882116, US15284411, US2008 / 017925A1, US2009 / 0014373A1, WO2012 / 058632

Claims

1. 1. An analyzer comprising: a frame (1) having an interface (2) for receiving an exchangeable cartridge (3) and fluidly connecting said exchangeable cartridge (3) to an analyzer for performing an analytical task; The replaceable cartridge (3) comprises: A liquid chromatography column (4, 5); a cartridge casing (6) for accommodating the liquid chromatography columns (4, 5); a heating block (7) thermally connected to the liquid chromatography columns (4, 5); A thermal interface (8), a heater (11) attached to the frame (1) included in the analyzer for heating the liquid chromatography columns (4, 5) of the replaceable cartridge (3); The heater (11) is movable between an open position for installing and removing the cartridge (3) in and from the interface (2) and a locked position for heating the liquid chromatography columns (4, 5).

2. 2. The analyzer of claim 1, wherein the heater (11) comprises a thermistor (15) for measuring temperature, the thermistor (15) being thermally connectable to the thermal interface (8) of the cartridge (3).

3. 3. The analyzer of claim 1 or 2, wherein the heater comprises a heat transfer part (13) movably mounted to the heater (11) to provide alignment with the thermal interface (8) of the cartridge (3) after installation of the cartridge.

4. 4. The analyzer according to claim 3, wherein the heater (11) comprises a pressing element (12) for pressing the heat transfer part (13) against the thermal interface (8).

5. An analyzer as described in any one of claims 1 to 4, wherein the heating block (7) has a surface extending outside the casing (6).

6. A liquid chromatography cartridge (3), comprising: a cartridge casing (6) for accommodating liquid chromatography columns (4, 5); a heating block (7) thermally connected to said liquid chromatography columns (4, 5); and a thermal interface (8) for thermally connecting said heating block to a heater (11) of an analyzer, The heater (11) is movable between an open position for placing the cartridge (3) on an interface (2) of the analyzer and a locked position for heating the liquid chromatography columns (4, 5) of the cartridge (3) placed on the interface (2).

7. 7. The liquid chromatography cartridge (3) of claim 6, wherein the thermal interface (8) is covered with a thermal tape (14).

8. 8. A liquid chromatography cartridge (3) according to claim 6 or 7, wherein the heating block (7) at least partially envelops and is set against a surface of the liquid chromatography columns (4, 5).

9. A liquid chromatography cartridge (3) described in any one of claims 6 to 8, wherein the heating block (7) has a surface extending outside the casing (6).

10. A method for heating a sample in a liquid chromatography column (4, 5) disposed in a casing (6) of a cartridge (3), comprising the steps of: - providing, inside said cartridge casing (6), a heating block (7) in contact with the surfaces of said columns (4, 5) and having a thermal interface (8); - bringing said heating block (7) into contact with a heat transfer element (13) of a heater (11) of an analyzer, said analyzer having an interface (2) for receiving said cartridge (3) and movable between an open position for placing said cartridge (3) on said interface (2) and a closed position for heating said liquid chromatography columns (4, 5) of said cartridge (3) placed on said interface (2); - heating said heating block (7) by said heater (11) for conducting thermal energy to said columns (4, 5).

11. - measuring the temperature of said heating block (7); The method of claim 10, further comprising: - controlling the temperature of the heating block (7) by means of the heater (11), thereby controlling the temperature of the samples in the columns (4, 5).

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