阅读说明：本技术 对流体和样品分离单元独立地进行热影响 (Independent thermal influence on fluid and sample separation units ) 是由 乔斯-安杰尔·莫拉 古韦·埃费尔斯贝格 莉娜·霍宁格 于 2021-06-18 设计创作，主要内容包括：本发明涉及对流体和样品分离单元独立地进行热影响,具体提供了一种用于样品分离设备(10)的热影响组件(100),该样品分离设备(10)用于通过样品分离单元(30)分离流动相中的流体样品,其中,热影响组件(100)包括热影响装置(80、82)和控制单元(70),热影响装置(80、82)配置为对流体样品和/或流动相和样品分离单元(30)进行热影响,控制单元(70)配置为控制热影响装置(80、82)来彼此独立地一方面对流体样品和/或流动相进行热影响而另一方面对样品分离单元(30)进行热影响。(The present invention relates to independently thermally influencing a fluid and a sample separation unit, in particular to a thermally influencing assembly (100) for a sample separation device (10), which sample separation device (10) is used for separating a fluid sample in a mobile phase by a sample separation unit (30), wherein the thermally influencing assembly (100) comprises thermally influencing means (80, 82) and a control unit (70), the thermally influencing means (80, 82) being configured to thermally influence the fluid sample and/or the mobile phase and the sample separation unit (30), the control unit (70) being configured to control the thermally influencing means (80, 82) to thermally influence the fluid sample and/or the mobile phase on the one hand and the sample separation unit (30) on the other hand independently of each other.)

1. A heat affected assembly (100) for a sample separation device (10), the sample separation device (10) being for separating a fluid sample in a mobile phase by a sample separation unit (30), wherein the heat affected assembly (100) comprises:

a heat influencing device (80, 82) configured to thermally influence the fluid sample and/or the mobile phase and the sample separation unit (30); and

a control unit (70) configured to control the heat influencing means (80, 82) to thermally influence the fluid sample and/or the mobile phase on the one hand and the sample separation unit (30) on the other hand independently of each other.

2. The heat affected assembly (100) according to claim 1, wherein the heat affected device (80, 82) comprises a first heat affected unit (80) configured to thermally affect the fluid sample and/or the mobile phase and comprises a second heat affected unit (82) configured to thermally affect the sample separation unit (30).

3. The heat-affected assembly (100) of claim 2, comprising at least one of the following features:

wherein the first heat-influencing unit (80) is thermally and/or functionally decoupled from the second heat-influencing unit (82);

wherein the control unit (70) is configured to separately control the first heat influencing unit (80) and the second heat influencing unit (82), in particular by separate control signals.

4. The heat affected assembly (100) according to any of claims 2 to 3, wherein the fluid sample and/or the mobile phase is controlled to be tempered by the first heat affected unit (80) and further by the second heat affected unit (82).

5. The heat-affected assembly (100) of claim 4, comprising at least one of the following features:

wherein the fluid sample and/or the mobile phase is arranged to be tempered directly by the first heat-influencing unit (80) and indirectly by the second heat-influencing unit (82);

wherein the fluid sample and/or the mobile phase are arranged to be heated by the second heat influencing unit (82) and selectively further heated or cooled by the first heat influencing unit (80).

6. The heat-affected assembly (100) according to any of claims 2 to 5, comprising at least one of the following features:

wherein the sample separation unit (30) is arranged to be tempered only by the second heat influencing unit (82);

wherein the first heat influencing unit (80) is arranged upstream of the second heat influencing unit (82);

wherein the first heat-influencing unit (80) and the second heat-influencing unit (82) are arranged in a spatially overlapping manner;

wherein the first heat influencing unit (80) is arranged within the second heat influencing unit (82);

wherein at least one of the first and second heat influencing units (80, 82) comprises at least one of the group consisting of a heatable or coolable mass, a Peltier element and a plasma heater;

wherein the second heat influencing unit (82) is configured to thermally influence the sample separation unit (30) without a gas convection directly acting on the sample separation unit (30).

7. The heat affected assembly (100) according to any of claims 2 to 6, wherein the second heat affected unit (82) is configured to thermally affect the sample separation unit (30) in case gas convection acts indirectly on the sample separation unit (30), in particular by providing:

a convection mechanism (96) for creating the gas convection (94) to facilitate thermal coupling of the sample separation unit (30); and

an at least partially thermally conductive shielding structure (88) shielding the gas convection (94) from the sample separation unit (30);

wherein optionally the at least partially thermally conductive shielding structure (88) comprises a heat exchanger (92), the heat exchanger (92) being configured to facilitate heat exchange between the gas convection (94) and the sample separation unit (30), and in particular to act as a heat source or sink.

8. The heat affected assembly (100) according to any one of claims 1 to 7, wherein the control unit (70) is configured to control the heat affected device (80, 82) such that the operation of the sample separation apparatus (10) emulates the operation of a further sample separation apparatus (110), in particular in terms of heat affecting the fluid sample and/or the mobile phase and in terms of heat affecting the sample separation unit (30), wherein preferably the control unit (70) is configured to emulate the operation of the further sample separation apparatus (110) based on the determined transfer function such that the sample separation apparatus (10) is in particular in terms of heat affecting the fluid sample and/or the mobile phase and in terms of heat affecting the sample separation unit (30) when a separation method developed for the further sample separation apparatus (110) is performed on the sample separation apparatus (10) The face behaves like the further sample separation device (110).

9. The heat affected assembly (100) according to any of claims 1 to 8, wherein the heat affected device (80, 82) is configured to heat, cool or selectively heat or cool the fluid sample and/or the mobile phase and/or the sample separation unit (30).

10. A sample separation device (10) for separating a fluid sample, the sample separation device (10) comprising:

a fluid driving unit (20) configured to drive a mobile phase and the fluid sample injected in the mobile phase;

a sample separation unit (30) configured to separate the fluid sample in the mobile phase; and

the heat affected assembly (100) according to any of claims 1 to 9 for heat affecting the fluid sample and/or the mobile phase on the one hand and the sample separation unit (30) on the other independently of each other.

11. The sample separation apparatus (10) according to claim 10, comprising a heat-affected compartment (84), the sample separation unit (30) being arranged in the heat-affected compartment (84).

12. The sample separation apparatus (10) according to claim 11, wherein a first heat influencing unit (80) configured to thermally influence the fluid sample and/or the mobile phase is arranged upstream of the heat influencing compartment (84).

13. The sample separation apparatus (10) according to any one of claims 10 to 12, comprising at least one further sample separation unit (30) connected in parallel to the sample separation unit (30) and comprising a selection valve (86) configured to select one of the sample separation units (30).

14. The sample separation apparatus (10) according to claim 12 or 13, comprising one of the following features:

wherein the first heat influencing unit (80) is integrated in the selector valve (86);

wherein the first heat influencing unit (80) comprises a metal microfluidic structure, in particular integrated in the selection valve (86);

wherein the first heat affected unit (80) is arranged between the selector valve (86) and the heat affected compartment (84);

wherein the first heat influencing unit (80) is arranged upstream of the selector valve (86).

15. The sample separation apparatus (10) according to claim 11, wherein a first heat influencing unit (80) configured to thermally influence the fluid sample and/or the mobile phase is arranged at least partially inside the heat influencing compartment (84), in particular thermally coupled to a head portion of the sample separation unit (30).

16. The sample separation device (10) according to any of claims 10 to 15, comprising a pre-processing assembly (90) for thermally pre-processing the fluid sample and/or the mobile phase upstream of the sample separation unit (30), wherein a first heat influencing unit (80) configured for thermally influencing the fluid sample and/or the mobile phase is thermally coupled to the pre-processing assembly (90).

17. The sample separation apparatus (10) according to any of claims 11 to 16, wherein a second heat influencing unit (82) configured to thermally influence the sample separation unit (30) is arranged at least partially inside the heat influencing compartment (84).

18. The sample separation apparatus (10) according to any one of claims 10 to 17, further comprising at least one of the following features:

the sample separation device (10) is configured as a chromatographic sample separation device, in particular a liquid chromatography sample separation device, a gas chromatography sample separation device or a supercritical fluid chromatography sample separation device;

an injector (40) configured to inject the fluid sample into the mobile phase;

a detector (50) configured to detect the separated fluid sample;

a fractionation unit (60) configured to collect the separated fluid sample;

a degassing device (27) for degassing at least a portion of the mobile phase.

19. A process of adjusting the temperature of a fluid sample and/or mobile phase and of a sample separation unit (30) in a sample separation device (10), wherein the process comprises:

for thermally influencing the fluid sample and/or the mobile phase and the sample separation unit (30); and

the thermal influence is controlled such that it influences the fluid sample and/or the mobile phase thermally on the one hand and the sample separation unit (30) thermally on the other hand independently of each other.

20. The process of claim 19, comprising at least one of the following features:

wherein the method comprises controlling a first heat influence unit (80) to thermally influence the fluid sample and/or the mobile phase independently of a thermal influence on the sample separation unit (30), and separately controlling a second heat influence unit (82) to thermally influence the sample separation unit (30) independently of a thermal influence on the fluid sample and/or the mobile phase;

wherein the method comprises controlling the thermal influence to perform a simulation of a separation method of a further sample separation device (110) by the sample separation device (10) such that the sample separation device (10) behaves like the further sample separation device (110), in particular with respect to the thermal influence on the fluid sample and/or the mobile phase and with respect to the thermal influence on the sample separation unit (30);

wherein the method comprises thermally influencing the fluid sample and/or the mobile phase by adjusting, in particular managing, the temperature of the fluid sample and/or the mobile phase, and/or comprises thermally influencing the sample separation unit (30) by adjusting, in particular managing, the temperature of the sample separation unit (30).

Technical Field

The present invention relates to a heat affected assembly, a sample separation device and a process for adjusting the temperature of a fluid sample and/or mobile phase and a sample separation unit in a sample separation device.

Background

In liquid chromatography, a fluid (such as a mixture between a fluid sample and a mobile phase, etc.) may be pumped through a conduit and a column comprising a material capable of separating different components of the fluid sample (the stationary phase). Such a material, i.e. so-called beads, which may comprise silica gel, may be packed into a column, which may be connected to other elements (such as a sampling unit, a flow cell, a container containing a sample and/or a buffer) via a conduit.

To operate the sample separation apparatus, the fluid may be preheated by a preheater assembly located downstream of the injector for injecting the fluid sample into the mobile phase and upstream of the column.

US 2015/0196855a1 discloses an apparatus for mounting components in a heating chamber of a fluid separation device for heating a fluid, wherein the apparatus comprises a mounting plate having at least one mounting recess, each mounting recess being configured to receive at least one component, and the at least one component each being configured to be mountable in and/or on the at least one mounting recess.

WO 2010/025777a1 discloses an apparatus for conducting an operating mode out of a first fluid device to a second fluid device, wherein the first fluid device has a first target operational mode representing a desired behavior of the first fluid device and has a first real operational mode representing an actual behavior of the first fluid device, wherein the second fluid device has a second target operational mode representing a desired behavior of the second fluid device and has a second real operational mode representing an actual behavior of the second fluid device, the device comprises a first determination unit adapted to determine a first real operation mode based on a first target operation mode and based on a pre-known parameterization of the first fluid device, and a second determination unit adapted to determine a second target operation mode based on the determined first real operation mode and based on a pre-known parameterization of the second fluid device.

US 2009/0076631a1 discloses an apparatus for determining an operation mode of a device, wherein the device is capable of adjusting physical conditions at a source location to correspondingly influence physical conditions at a destination location, the apparatus comprising a determining unit adapted to determine the operation mode by defining a time dependency of the physical conditions at the source location to obtain a target time dependency of the physical conditions at the destination location, the target time dependency representing a resultant change of the physical conditions over time.

Disclosure of Invention

The object of the present invention is to enable a sample separation device for separating a fluid sample in a mobile phase to be operated in a flexible manner. This object is solved by the independent claims. Further embodiments are shown by the dependent claims.

According to an exemplary embodiment of the present invention, there is provided a heat-affected assembly for a sample separation apparatus, wherein the heat-affected assembly includes: a heat influencing device configured to thermally influence the fluid sample to be separated and/or a mobile phase through which the fluid sample may be transported or in which the fluid sample is transported, and to thermally influence the sample separation unit; and a control unit configured to control the heat-influencing device to influence the heat on the fluid sample and/or the mobile phase on the one hand and the sample separation unit on the other hand independently of each other.

According to another exemplary embodiment of the present invention, a sample separation device for separating a fluid sample is provided, wherein the sample separation device comprises a fluid driving unit configured to drive a mobile phase and the fluid sample injected in the mobile phase, a sample separation unit configured to separate the fluid sample in the mobile phase, and a heat influencing assembly having the above mentioned features for thermally influencing the fluid sample and/or the mobile phase on the one hand and the sample separation unit on the other hand independently of each other.

According to a further exemplary embodiment, a process for adjusting the temperature of a fluid sample and/or a mobile phase and of a sample separation unit in a sample separation device is provided, wherein the process comprises: thermally influencing the fluid sample and/or the mobile phase and the sample separation unit; and controlling the thermal influence so as to influence the thermal influence on the fluid sample and/or the mobile phase on the one hand and on the sample separation unit on the other hand independently of each other.

In the context of the present application, the term "sample separation device" may particularly denote any device capable of separating different components of a fluid sample by applying a certain separation technique, in particular liquid chromatography.

In the context of the present application, the term "fluid sample" may particularly denote any liquid and/or gaseous medium to be analyzed, optionally also comprising solid particles. Such a fluid sample may comprise various fractions of molecules or particles to be separated, e.g. small mass molecules or large mass biomolecules such as proteins etc. The separation of a fluid sample into fractions may involve certain separation criteria (such as mass, volume, chemical properties, etc.) according to which the separation is performed.

In the context of the present application, the term "mobile phase" may particularly denote any liquid and/or gaseous medium that may be used as a fluid carrier for a fluid sample during separation. The mobile phase may be a solvent or a combination of solvents (e.g., consisting of water and an organic solvent such as ethanol or acetonitrile, etc.). In an isocratic separation mode of a liquid chromatography apparatus, the mobile phase may have a composition that is constant over time. However, in the gradient mode, the composition of the mobile phase may change over time, in particular so as to desorb a fraction of the fluid sample that has previously been adsorbed to the stationary phase of the sample separation unit.

In the context of the present application, the term "fluid driving unit" may particularly denote an entity capable of driving a fluid (i.e. a liquid and/or a gas, optionally comprising solid particles), in particular a fluid sample and/or a mobile phase. For example, the fluid driver may be a pump (e.g., in the form of a piston pump or peristaltic pump) or other high pressure source. For example, the fluid driver may be a high pressure pump, e.g. capable of driving the fluid at a pressure of at least 100bar, in particular at least 500 bar.

The term "sample separation unit" may particularly denote a fluidic member through which a fluid sample is transferred and which is configured such that upon guiding the fluid sample through the separation unit, the fluid sample will be separated into different groups of molecules or particles. One example of a separation unit is a liquid chromatography column, which is capable of collecting or retaining and selectively releasing different fractions of a fluid sample.

The term "heat influencing assembly" may particularly denote a device configured to thermally influence or temper the fluid (in the form of the fluid sample and/or the mobile phase) and the sample separation unit. Thermal influence, thermal change or thermal manipulation may mean in particular a change of the temperature in a controlled or even regulated manner. In particular, the thermal influence may be achieved by heating (i.e. by supplying thermal energy) and/or by cooling (i.e. by removing thermal energy).

The term "heat influencing means" may particularly denote a means which may be suitably controlled to thermally influence the fluid and the sample separation unit, respectively. Such a heat-influencing device may comprise a plurality of heat-influencing units, each of which is separately controlled by the control unit and each of which is capable of heating or cooling a respectively assigned destination. The destination may for example be a fluid (in particular a fluid sample or a mobile phase) which may for example be heated while flowing through the conduit or while being surrounded by the pre-heater assembly. The destination may also be a sample separation unit, which may be directly heated or cooled, e.g. when arranged in a compartment.

The term "independently controlling the thermal influence" may particularly denote that a controlled thermal influence on the fluid sample and/or the mobile phase may be performed without the process being required to be forcibly limited or influenced by another process for controlled thermal influence on the sample separation unit. Thus, one thermal influence can be controlled regardless of the other thermal influence. Although the results of the controlled tempering process may have a certain influence on each other due to thermal interactions between the mobile phase and/or the fluid sample flowing through the sample separation unit, the external adjustments to the two heat affected processes may be performed separately or independently from each other, e.g. using different control signals for the two heat affected processes. Thus, there may be independence on the control side.

According to an exemplary embodiment, the temperature control of the mobile phase and/or the fluid sample on the one hand and the temperature control of the sample separation unit for separating the fluid sample on the other hand may be decoupled from each other on the control side. By taking this measure, an additional degree of freedom or additional design parameters may be provided compared to the case of preheating the fluid sample, the mobile phase and the sample separation unit by controlling all mentioned elements in the same way by one common control process. According to an exemplary embodiment, a functional separation or independent configuration on the control side with a thermal influence on the fluid on the one hand and on the sample separation unit on the other hand may allow an improved or more precise pre-heating (or more generally: thermal conditioning) in the sample separation. Furthermore, the independent adjustability of the fluid temperature and the temperature of the sample separation unit may be a highly suitable basis for transferring separation methods developed for conventional sample separation devices to another sample separation device according to an exemplary embodiment of the present invention. The operating parameters for independent temperature adjustment of the fluid and sample separation unit may be adjusted such that a sample separation apparatus according to an exemplary embodiment may be flexibly configured and reconfigured to behave like many different conventional sample separation apparatuses in terms of temperature management. This may enable operating the sample separation apparatus according to exemplary embodiments of the present invention to separate a fluid sample in a mobile phase in a highly flexible manner.

Further embodiments of the heat affected assembly, sample separation apparatus and process will be explained below.

In an embodiment, the heat influencing device comprises a first heat influencing unit (which may be operable independently of a second heat influencing unit described below) configured to thermally influence the fluid sample and/or the mobile phase, and comprises a second heat influencing unit (which may be operable independently of the first heat influencing unit) configured to thermally influence the sample separation unit. The heat-influencing units may operate independently of each other. The two structurally separate heat influencing units may form a suitable hardware basis for functionally independent temperature adjustment of the fluid and sample separation units separately. At least one third heat influencing unit may also be provided in order to further improve the tempering and/or to further increase the degree of freedom to emulate the separation behavior of another conventional sample separation device by means of a sample separation device according to an exemplary embodiment of the present invention.

In an embodiment, the first heat influencing unit is thermally decoupled from the second heat influencing unit. Such thermal decoupling may be achieved, for example, by sandwiching a thermally insulating material between the first and second heat-influencing units. The mentioned thermal decoupling may facilitate a functional decoupling between the preheating of the fluid and the preheating of the sample separation unit prior to the sample separation process.

In an embodiment, the control unit is configured to control the first and second heat influencing units separately. In particular, this may be achieved by supplying different and independent control signals from the control unit to the first heat influencing unit on the one hand and to the second heat influencing unit on the other hand.

In an embodiment, the fluid sample and/or the mobile phase is arranged to be tempered by the first heat influencing unit and additionally by the second heat influencing unit. In particular, the fluid sample and/or the mobile phase may be arranged to be directly tempered by the first heat influencing unit and indirectly tempered by the second heat influencing unit. For example, the fluid sample and/or the mobile phase may be arranged to be heated by a second heat influencing unit (e.g. a heated heating plate or other mass) and selectively further heated or cooled by a first heat influencing unit (e.g. in the form of a peltier unit). For example, the sample separation unit may be arranged to be tempered only by the second heat influencing unit. Such an embodiment is shown, for example, in fig. 7. In such a configuration, it is for example possible that a large part of the thermal energy provided for thermally influencing or changing both the fluid and the sample separation unit is provided by a sufficiently strong second thermal influencing unit which directly heats the sample separation unit and indirectly heats the fluid. The first heat influencing unit may then be used for improving the temperature control of the fluid, i.e. may be configured to be small and accurate.

Alternatively, it is also possible to directly thermally influence or regulate (in particular heat) the fluid by only one thermal influencing unit, while the sample separation unit may be tempered by both the first thermal influencing unit and the second thermal influencing unit.

In an embodiment, the first heat influencing unit is arranged partially or completely upstream (in the flow direction of the mobile phase and the fluid sample) of the second heat influencing unit. In other words, the preheating of the fluid may be performed before the fluid reaches the sample separation unit.

In an embodiment, the first and second heat influencing units are arranged in a spatially overlapping manner. Alternatively, the first heat-influencing unit may be arranged entirely within (i.e., inside) the second heat-influencing unit. In both configurations, it is possible, for example, that the second heat-affected cell may heat the entire sample separation cell(s), while the first heat-affected cell thermally controls only a portion (preferably the head portion) of the sample separation cell(s).

In an embodiment, at least one of the first and second heat influencing units comprises at least one of the group consisting of a heatable or coolable mass (such as a heating plate or the like), a peltier element and a plasma heater. Heating or cooling the block may be achieved, for example, by cooling a liquid (such as cold water or the like) or heating a liquid (such as hot water or the like). Heating the mass may also be achieved by ohmic heating, i.e. by applying a current that heats the mass through ohmic losses. The peltier element may be a thermoelectric cooler comprising different semiconductors in contact with each other, wherein the application of a current causes heating or-when the direction of the current is reversed-causes cooling. The plasma heater may, for example, be an arc heater, which may be a low temperature plasma generator, wherein an arc discharge acts as a heat release element. Plasma heating can also be used in the manufacturing process of ohmic heaters, as it allows the deployment of a sandwich structure of a mixture of dielectric and conductive layers in, for example, a planar structure, such as a metal microfluidic structure, thereby achieving high energy density in a small space.

In an embodiment, the second heat influencing unit is configured to thermally influence the sample separation unit without causing a convective gas flow to impinge on the sample separation unit via a direct gas flow directed onto the sample separation unit. Avoiding such gas flows to directly affect the sample separation unit may improve the separation performance, in particular the chromatographic separation performance, of the sample separation unit(s). In one aspect, gas flow or gas convection is a powerful mechanism to facilitate heat exchange. On the other hand, it has been demonstrated that applying a gas flow directly to a sample separation unit (such as a chromatography separation column or the like) in terms of heating may result in a significant temperature distribution over the radial extension of the sample separation unit. This may deteriorate the separation performance. It has been found that excellent results in terms of pre-heating and separation performance can be achieved by using indirect gas convection to facilitate heat exchange while protecting the sample separation unit from direct impact of the gas convection.

In an embodiment, the second heat influencing unit is configured to thermally influence the sample separation unit in case gas convection acts only indirectly on the sample separation unit. This may be achieved, for example, by providing a convection mechanism for generating a gas convection to facilitate thermal coupling of the sample separation unit and an at least partially thermally conductive shielding structure shielding or mechanically spacing the gas convection from the sample separation unit. An air flow mechanically decoupled but thermally coupled to the sample separation unit may provide improved temperature stability, enhanced environmental rejection and rapid thermal equilibrium, while achieving high separation performance. Illustratively, gas convection acting only indirectly on the sample separation unit may facilitate thermal coupling of the sample separation unit during operation and increase thermal uniformity. Optionally but advantageously, the at least partially thermally conductive shielding structure comprises a heat exchanger configured to promote heat exchange between the convective gas flow and the sample separation unit. For example, the heat exchanger may also be used as a heat source (i.e., heat may be supplied for heating) or a heat sink (i.e., heat may be removed for cooling). In such embodiments, the one or more sample separation cells may be partially or completely surrounded by a shielding structure that shields gas convection from directly impacting the sample separation cell(s). At the same time, the convection of gases around the outer surface of the shielding structure (and preferably inside a heat-affected compartment or cavity such as a column furnace or the like) may also facilitate heat exchange inside the shielding structure and may thus have a positive influence on the thermal controllability of the sample separation unit(s). In an embodiment, the actual heating or cooling source may form part of a heat exchanger.

In an embodiment, the control unit is configured to control the heat influencing means such that the operation of the sample separation device emulates the operation of another sample separation device. In particular, such an emulation may be performed in terms of a thermal influence on the fluid sample and/or the mobile phase and in terms of a thermal influence on the sample separation unit. Advantageously, the additional degree of freedom or increased number of design parameters in the form of two (rather than one) tempering entities may allow for adjusting the tempering parameters such that the heat affected assembly of the sample separation apparatus according to an exemplary embodiment of the present invention behaves like a heat affected assembly of a conventional or another sample separation apparatus when performing the separation method.

In an embodiment, the emulation of the tempering behavior of the further sample separation device may be combined with an emulation of the further sample separation device with respect to at least one further aspect, in particular with respect to a time-dependent emulation of a solvent composition of the mobile phase during sample separation. For example, the behavior of the further sample separation device may be emulated by the sample separation device according to an exemplary embodiment of the present invention with respect to the gradient profile during gradient run.

In an embodiment, the control unit is configured to emulate the operation of the further sample separation device based on the determined transfer function (e.g. by the control unit) such that the sample separation device behaves like the further sample separation device, in particular with respect to a thermal influence on the fluid sample and/or the mobile phase and with respect to a thermal influence on the sample separation unit, when a separation method developed for the further sample separation device is performed on the sample separation device. In the context of the present application, the term "separation method" may particularly denote an instruction for a sample separation device on how to separate a fluid sample, which instruction is to be performed by the sample separation device to perform a separation task associated with the separation method. Such a separation method may be defined by a set of parameter values (e.g. temperature, pressure, properties of the solvent composition, etc.) and hardware components of the sample separation apparatus (e.g. the type of separation column used) and an algorithm with the processes performed when performing the separation method. A corresponding set of technical parameters for operating the sample separation apparatus during sample separation may be known, for example stored in a database or memory accessible by a control unit controlling the operation of the sample separation apparatus. The physical properties or operating parameters characterizing the separation method may comprise transport characteristics, which may comprise values of physical parameters like volume, dimensions, like pressure or temperature, and/or physical effects like a friction model occurring in the fluid conduit (which may be modeled, for example, according to the harbourne's law), etc. More particularly, the parameterization may take into account the size of the sample separation device (e.g. the size of the fluid channel), the volume of the fluid conduit of the sample separation device (such as the dead volume, etc.), the pump performance of the sample separation device (such as the pump power and/or pump capacity, etc.), a delay parameter for operating the sample separation device (such as the delay time after switching on the sample separation device, etc.), a friction parameter for operating the sample separation device (e.g. characterizing the friction between the wall of the fluid conduit and the fluid flowing through the conduit), the flushing performance of the sample separation device (in particular the performance related to cleaning or flushing the sample separation device before operating the sample separation device or between two consecutive operations), and/or the cooperation of different components of the sample separation device (e.g. the performance of the gradient applied to the chromatography column). By calculating a transfer function that can be applied to transfer a separation method developed for a conventional sample separation apparatus to be used by the sample separation apparatus according to an exemplary embodiment of the present invention, a numerically simple manner of transferring a separation method from one sample separation apparatus to another sample separation apparatus can be accomplished.

In an embodiment, the sample separation apparatus comprises a heat-affected compartment in which the at least one sample separation unit is arranged. Such a heat-affected compartment may be a column oven for preheating fluid and sample separation unit(s) in preparation for sample separation.

In an embodiment, the above-mentioned first heat influencing unit configured to thermally influence the fluid sample and/or the mobile phase is located upstream of the heat influencing compartment. The described geometrical configuration may further contribute to a proper functional separation between the first and second heat influencing units when the second heat influencing unit is arranged inside the heat influencing compartment.

In an embodiment, the sample separation device comprises at least one further sample separation unit connected in parallel with the above-mentioned sample separation unit, and comprises a fluid selection valve configured to select one of the sample separation units. Preferably, the first heat influencing unit may be integrated in the selection valve. This configuration is highly compact as it allows for thermal influence on the fluid before it is divided into a plurality of paths, wherein each path comprises one of the sample separation units. At the same time, this configuration may ensure that preheating takes place spatially close to the location of the sample separation unit for separating the fluid sample.

In an embodiment, the first heat influencing unit is configured as a metal microfluidic structure, in particular integrated in the selection valve. In particular, the Metal Microfluidic (MMF) heater may advantageously be integrated into a fluid selection valve, which may also be referred to as a channel selection valve. Microfluidics focuses on the behavior of small-sized liquids and gases, which may be very different from that of macroscopic fluids, since at this scale negligible effects at the macroscopic scale may dominate. The fluid selection valve may be produced based on a metal structure which may be manufactured by thermal bonding from a stainless steel foil at high pressure and temperature. Thus, the channel select valve may be heated or cooled to complete the valve temperature control. In particular, a pre-column liquid regulator (in particular a heater and/or cooler) may be provided, which may be embedded in the column selection valve. In other words, heating and/or cooling capacity may be integrated into the selection valve.

In an embodiment, the first heat affected unit is arranged between the selector valve and the heat affected compartment. This may allow preheating of the fluid very close to the separation location in the sample separation unit.

In an embodiment, the first heat influencing unit is arranged upstream of the selector valve. The selection of the desired sample separation unit can then be made using the already preheated fluid.

In an embodiment, the first heat influencing unit configured to thermally influence the fluid sample and/or the mobile phase is at least partially arranged inside the heat influencing compartment, in particular thermally coupled to a head portion of the sample separation unit. The head portion of the sample separation unit may be the portion at which the mobile phase and the fluid sample enter the sample separation unit during a separation run. Such a configuration allows for a suitable pre-heating of the fluid sample and/or the mobile phase, especially at the separation location. Thus, no significant undesired cooling of the preheated sample due to temperature equilibration phenomena occurs in such embodiments.

In an embodiment, the sample separation device comprises a pre-processing assembly for thermal pre-processing (in particular pre-heating) the fluid sample and/or the mobile phase upstream of the sample separation unit, wherein a first heat influencing unit configured for thermally influencing the fluid sample and/or the mobile phase is thermally coupled to the pre-processing assembly. The pre-treatment assembly may be a thermally conductive structure surrounding a fluid-carrying conduit for promoting uniform heating of the fluid by the first heat influencing unit. Illustratively, the first heat-influencing unit may supply or remove thermal energy distributed by the pre-processing assembly along the fluid-carrying conduit.

In an embodiment, a second heat influencing unit configured for heat influencing the sample separation unit is arranged at least partially inside the heat influencing compartment. The second heat influencing unit may be arranged downstream of the first heat influencing unit.

In an embodiment, the fluid sample and/or the mobile phase may be tempered by adjusting, in particular managing, the temperature of the fluid sample and/or the mobile phase. Accordingly, the sample separation unit may be thermally influenced by adjusting, in particular managing, the temperature of the sample separation unit. Thus, the heat influencing unit may be configured to bring the fluid and the sample separation unit to respective target temperatures.

The sample separation unit may be filled with a separation material. Such a separation material, which may also be referred to as a stationary phase, may be any material that allows an adjustable degree of interaction with a fluid sample so as to be able to separate different components of such a sample. The separation material may be a liquid chromatography column packing material or packing material comprising at least one of the group consisting of: polystyrene, zeolite, polyvinyl alcohol, polytetrafluoroethylene, glass, polymer powder, silica and silica gel, or any of the above materials with a chemically modified (coated, capped, etc.) surface. However, any packing material having material properties that allow separation of analytes passing through the material into different components (e.g., due to differences in the type or affinity of interaction between the packing material and the analyte fraction) may be used.

At least a portion of the sample separation unit may be filled with a fluid separation material, wherein the fluid separation material may comprise beads having a size in a range of substantially 1 μm to substantially 50 μm. Thus, the beads may be small particles that may be packed inside the separation section of the microfluidic device. The beads may have pores of a size in the range of substantially 0.01 μm to substantially 0.2 μm. The fluid sample may pass through the cavity, wherein an interaction may occur between the fluid sample and the cavity.

The sample separation unit may be a chromatography column for separating components of the fluid sample. Thus, exemplary embodiments may be implemented particularly in the context of a liquid chromatography apparatus.

The fluid separation system may be configured to direct the liquid mobile phase through the separation cell. As an alternative to a liquid mobile phase, a fluid separation system may be used to treat a gaseous mobile phase or a mobile phase comprising solid particles. The exemplary embodiments can also be used to process materials that are mixtures of different phases (solid, liquid, gas). The sample separation device, in particular the fluid driving unit thereof, may be configured to direct the mobile phase through the system at a high pressure, in particular at a high pressure of at least 600bar, more in particular at least 1200 bar.

The sample separation apparatus may be configured as a microfluidic device. The term "microfluidic device" may particularly denote a sample separation apparatus as described herein, which allows for transporting a fluid through a microchannel having a size in the order of magnitude of less than 500 μm, in particular less than 200 μm, more in particular less than 100 μm or less than 50 μm or less.

The exemplary embodiments may be implemented by a sample injector of a liquid chromatography apparatus that may take a fluid sample from a fluid container and may inject such a fluid sample into a conduit for supply to a separation column. During such a process, the fluid sample may be compressed from, for example, a normal pressure to a higher pressure, for example, several hundred bar or even 1000bar or more. The autosampler may automatically inject a fluid sample from a vial into the sample loop. The tip or needle of the autosampler may be immersed in a fluid container, fluid may be drawn into a capillary tube, then may be driven back into the seat, and then a fluid sample may be injected toward a sample separation section of the liquid chromatography apparatus, e.g., via a switchable fluid valve.

The sample separation device may be configured to analyze at least one physical, chemical and/or biological parameter of at least one component of the fluid sample in the mobile phase. The term "physical parameter" may particularly denote the size or temperature of the fluid. The term "chemical parameter" may particularly denote the concentration of a fraction of the analyte, an affinity parameter, etc. The term "biological parameter" may particularly denote the concentration of proteins, genes etc., the biological activity of a component etc. in a biochemical solution.

The sample separation apparatus may be implemented in various technical environments, such as a sensor device, a testing device, a device for chemical, biological and/or pharmaceutical analysis, a capillary electrophoresis device, a liquid chromatography device, a gas chromatography device, an electronic measurement device or a mass spectrometry device. In particular, the sample separation device may be a High Performance Liquid Chromatography (HPLC) apparatus by which different fractions of an analyte may be separated, examined and analyzed.

Embodiments of the invention include a sample separation device configured to separate a complex of a fluid sample in a mobile phase. The sample separation device comprises a mobile phase driver, such as a pumping system or the like, configured to drive the mobile phase through the sample separation device. A sample separation unit, which may be a chromatography column, is arranged to separate complexes of the sample fluid in the mobile phase. The sample separation device may further comprise a sample injector configured to introduce the fluid sample into the mobile phase, a detector configured to detect the separated complexes of the fluid sample, a collector configured to collect the separated complexes of the fluid sample, a control unit or data processing unit configured to process data received from the sample separation device, and/or a degassing device for degassing the mobile phase.

Embodiments of the present invention may be presented based on commonly available HPLC systems such as agilent 1290 series infinite systems, agilent 1200 series fast resolution LC systems, or agilent 1100HPLC series (all provided by applicant agilent technologies-see www.agilent.com-which should be incorporated herein by reference).

One embodiment includes a pumping apparatus having a piston for reciprocating in a pump working chamber to compress a liquid in the pump working chamber to a high pressure at which the rate of compression of the liquid becomes apparent. One embodiment comprises two pumping devices coupled in series (as disclosed for example in EP309596a 1) or in parallel.

The mobile phase (or eluent) can be a pure solvent or a mixture of different solvents. The choice may be made, for example, to minimize the retention of the target complex and/or the amount of mobile phase to run the chromatography. The mobile phase may also be chosen such that different complexes can be separated efficiently. The mobile phase may comprise organic solvents like methanol or acetonitrile, often diluted with water. For gradient runs, water and organics are delivered in separate bottles, and a gradient pump delivers the planned blend from the bottle to the system. Other commonly used solvents may be isopropanol, THF, hexane, ethanol and/or any combination thereof, or any combination of these with the aforementioned solvents.

The fluid sample may comprise any type of process liquid, a natural sample such as blood plasma, if juice, or it may be the result of a reaction such as from a fermentation broth.

The fluid is preferably a liquid, but may also be or include a gas and/or a supercritical fluid (as used in supercritical fluid chromatography-SFC as disclosed, for example, in US4,982,597A).

Drawings

Other objects and many of the attendant advantages of this invention will be readily appreciated as the same become better understood by reference to the following more detailed description of the embodiments, when taken in conjunction with the accompanying drawings. Features that are substantially or functionally equivalent or similar will be referred to by the same reference numerals.

Fig. 1 shows a sample separation apparatus according to an embodiment of the present invention, in particular for High Performance Liquid Chromatography (HPLC), wherein the thermal influence on the fluid sample in the mobile phase is performed independently of the thermal influence on the sample separation unit for separating the fluid sample.

Fig. 2 is a schematic diagram of a heat-affected assembly for a sample separation apparatus according to an exemplary embodiment, wherein a first heat-affected unit is integrated in a selection valve and a second heat-affected unit is arranged inside a heat-affected compartment.

Fig. 3 is a schematic diagram of a heat-affected assembly for a sample separation apparatus according to an exemplary embodiment, wherein a first heat-affected unit is disposed in a head portion of the sample separation unit and a second heat-affected unit is disposed inside a heat-affected compartment.

Fig. 4 is a schematic diagram of a heat-affected assembly for a sample separation apparatus according to an exemplary embodiment, wherein a first heat-affected unit is disposed between a selection valve and a heat-affected compartment, and a second heat-affected unit is disposed inside the heat-affected compartment.

Fig. 5 is a schematic diagram of a heat-affected assembly for a sample separation apparatus according to an exemplary embodiment, wherein a first heat-affected unit is disposed upstream of a selection valve and a second heat-affected unit is disposed inside a heat-affected compartment.

Fig. 6 is a schematic diagram of a heat-affected assembly for a sample separation apparatus according to an exemplary embodiment, wherein a first heat-affected unit is disposed inside the heat-affected compartment and a second heat-affected unit is also disposed inside the heat-affected compartment but downstream of the first heat-affected unit.

Fig. 7 is a schematic view of a heat influencing assembly for a sample separation apparatus according to an exemplary embodiment, wherein a first heat influencing unit for thermally influencing only the fluid sample and/or the mobile phase in the pre-treatment assembly and a second heat influencing unit for thermally influencing the fluid sample and/or the mobile phase and for thermally influencing the sample separation unit in the heat influencing compartment are provided.

FIG. 8 is a schematic view of a portion of a heat-affected assembly in a heating compartment of a sample separation device, wherein the sample separation unit is heated by a convection mechanism that operates only indirectly, according to an exemplary embodiment.

FIG. 9 is a schematic diagram of a heat-affected component of a sample separation device according to an exemplary embodiment, wherein operation of the heat-affected component emulates the temperature-regulating behavior of another sample separation device.

Fig. 10 is a three-dimensional view of a heat-affected unit (or a portion thereof) for a heat-affected assembly of a sample separation device according to an exemplary embodiment, wherein the heat-affected unit is configured as a metal microfluidic structure for heating or cooling a mobile phase and/or a fluid sample and is provided integrated in a channel selection valve.

The illustration in the drawings is schematically.

Detailed Description

Before referring to the drawings, exemplary embodiments will be explained in more detail, some basic considerations will be explained based on the exemplary embodiments that have been developed.

According to an exemplary embodiment of the present invention, a heat affected component (such as a sample in a mobile phase and a separation column pre-heater) for a sample separation apparatus (such as a liquid chromatography apparatus or the like) is provided which enables tempering (in particular temperature control or temperature adjustment) of a fluid sample to be separated and/or a mobile phase for carrying the fluid sample on the one hand and a sample separation unit (such as a chromatography separation column or the like) on the other hand. In other words, the preheating of the sample/mobile phase may be done independently of the preheating of the sample separation unit for separating the sample. The gist of an exemplary embodiment is thus to use separate heating sources for heating the mobile phase (which may be performed in a preheater) on the one hand and the separation column on the other hand.

Typically, the preheater and the separation column may be tempered together, for example via a common heating block and by implementing one or more heat exchangers. According to an exemplary embodiment, it may be advantageous that the heating source for the mobile phase and the sample is separated with respect to the heating source of the sample separation unit. In particular, it may be advantageous that by separating the heating source, other column furnace types (or more generally other heat affected compartments) may be emulated or simulated. By this positive concept with two heat influencing sources, it is possible to simulate another column furnace with the negative concept of only one heating source. Illustratively, the functional and logical separation between the mobile phase tempering in the sample separation device and the tempering of the sample separation unit provides an additional degree of freedom, which can be used as a design parameter for simulating the operation of another sample separation device by enabling a thermal influence on the mobile phase/fluid sample and the sample separation unit(s) independently of each other. For example, the operation of an independently adjustable tempering mechanism of a sample separation device according to an exemplary embodiment of the present invention may be set to mimic, emulate or simulate the function of another sample separation device in terms of preheating.

In an advantageous embodiment, the heat affected compartment for heat affecting the one or more sample separation units (which may also be referred to as a column compartment) may be regulated by two independently controlled heat affected units (which may be heaters and/or coolers), one dedicated to regulating the liquid temperature of the fluid sample and/or mobile phase, the other regulating the temperature inside the heat affected compartment (and thereby regulating the temperature of the one or more sample separation units in the heat affected compartment).

When designing a column compartment according to an exemplary embodiment of the present invention, it may be advantageous to achieve reproducible operating conditions of the column, maintaining backward compatibility with existing separation methods operating in other instruments (e.g. conventional instruments). Maintaining backward compatibility may have an impact on improving the performance of the new model. In general, when a separation method developed for one sample separation device is run on another sample separation device, the same performance may not be exhibited under the same operating conditions (such as flow rates and/or temperatures of mobile phase and fluid sample, gradients associated with changing solvent composition of the mobile phase, etc.) in the new sample separation device, which may be a disadvantage. To overcome these disadvantages, exemplary embodiments of the present invention may use two independently controlled heat affected units (such as heaters and/or coolers, etc.) to adjust a heat affected compartment (particularly a chromatography column compartment). In this case, one of the heat influencing units may be dedicated to regulating the liquid temperature of the mobile phase and/or the fluid sample, and the other heat influencing unit may be arranged to regulate the temperature inside the heat influencing compartment. Advantageously, such an embodiment may ensure backward compatibility and may improve separation performance.

Thus, independent or separate control of the thermal influence on the mobile phase and the fluid sample on the one hand and the thermal influence on one or more sample separation units of the sample separation device on the other hand may make the sample separation device backwards compatible and adaptable to conventional separation methods. Furthermore, taking such measures may allow the design of sample separation devices that achieve significant improvements in performance. Furthermore, the use of independent heat influencing units (which may comprise independently controllable heating and/or cooling units) for the liquids (i.e. the mobile phase and the fluid sample) may reduce the number of pre-column heaters to one, thereby reducing hardware effort. Thus, providing separate or independent heat influencing units for the mobile phase and the fluid sample may increase the flexibility of operation. For example, such independently controllable heat-affected units (i.e., pre-column heaters and/or coolers) may be integrated into the selection valve, for example using one or more peltier coolers and/or one or more plasma heaters. Such a selection valve may be configured to select one of a plurality of parallel connected sample separation units, for example, according to the requirements of a particular application. Because such a selection valve may be arranged directly upstream of the sample separation unit and thus directly upstream of the heat-affected compartment, the independent control or adjustment of the temperature of the mobile phase and the fluid sample may be spatially very close to the adjustment of the temperature of the sample separation unit in the heat-affected compartment. Thus, the undesired temperature equilibration process may be kept small without compromising the independent adjustability of the temperature regulation characteristics of the fluid and sample separation unit.

Thus, exemplary embodiments of the present invention may enable thermal conditioning of the liquid prior to entering the heat-affected compartment having the sample separation unit(s) therein, which may avoid internal condensation problems and temperature instability.

Exemplary embodiments of the present invention may incorporate a first heat influencing unit (which may be a heater and/or a cooler) that brings the temperature of the liquid (i.e., the mobile phase and the fluid sample) to a set point. A second heat influencing unit (which may also be a heater and/or a cooler) may be arranged to independently control the temperature of the heat influencing compartment (comprising one or more sample separation units), e.g. by means of control logic to achieve optimal performance of the separation as a separation degree of freedom that may be used for developing separation methods. Furthermore, this may make it possible to make the heat affected compartment backwards compatible with conventional sample separation methods and/or conventional sample separation devices. For example, the pre-column conditioner in the form of an independently controllable first heat influence unit may be located inside or outside a section in which one or more chromatography separation columns are assigned and in which heat influence by the second heat influence unit may occur.

Another aspect of exemplary embodiments of the present invention is an HPLC column furnace having a hybrid configuration with respect to gas convection, i.e., a hybrid configuration with and without air circulation. In particular, a column compartment may be provided which is conditioned by an air flow directed around the column area. In conventional HPLC column compartments, no active air flow is provided at all (resulting in a more adiabatic environment), or forced air flow may be provided to the compartment (resulting in a more isothermal environment). In contrast to these approaches, a post compartment according to an exemplary embodiment of the present invention may provide a forced air flow around the area of the positioning post, while the forced air flow at the position of the post itself may be reliably prevented, e.g. by shielding. It has been shown that compartments with low (i.e. no forced) air flow around the column may allow to obtain better chromatographic results. Forced rapid air flow can result in better temperature stability, better environmental phenomenon suppression, and faster equilibration. According to an exemplary embodiment of the invention, the forced air flow may be directed around the post location by the flow redirector shield, but is preferably not directed to the post location. Due to the smaller temperature difference, the area around the column may have a significantly reduced air flow. This may result in higher temperature stability, may reduce the need for thick isolation, and may retain good chromatographic results.

Referring now in more detail to the drawings, FIG. 1 depicts a general schematic diagram of a liquid separation system as an example of a sample separation apparatus 10 according to an exemplary embodiment of the present invention. As will be described in further detail below, embodiments include performing a thermal influence on the fluid sample in the mobile phase independently of a thermal influence on the sample separation unit 30 for separating the fluid sample.

The pump or fluid drive unit 20 typically receives the mobile phase from the solvent supply 25 via a degasser 27, the degasser 27 degassing and thus reducing the amount of gas dissolved in the mobile phase. The fluid driving unit 20 drives a mobile phase through a sample separation unit 30 (such as a chromatography column, etc.) including a stationary phase. A sampling unit or injector 40 may be disposed between the fluid drive unit 20 and the sample separation unit 30 in order to inject or add (often referred to as sample introduction) the sample fluid or fluid sample into the mobile phase. The stationary phase of the sample separation unit 30 is configured to separate a complex of sample liquids. The detector 50 is arranged for detecting the separated complexes of the sample fluid. A fractionation unit 60 may be provided for outputting the separated complexes of the sample fluid. The separated sample fluid and mobile phase composite may also be transported to a waste line (not shown).

Although the mobile phase may consist of only one solvent, it may also be a mixture of a plurality of solvents. This mixing may be a low pressure mixing and is arranged upstream of the fluid drive unit 20 such that the fluid drive unit 20 has received and pumped the mixed solvent as a mobile phase. Alternatively, the fluid drive unit 20 may be comprised of a plurality of separate pumping units, wherein the plurality of pumping units each receive and pump a different solvent or mixture such that mixing of the mobile phase (received by the sample separation unit 30) occurs at high pressure and downstream of (or as part of) the fluid drive unit 20. The composition (mixture) of the mobile phase may be kept constant over time (so-called isocratic mode) or may vary over time (so-called gradient mode).

A data processing unit or control unit 70, which may be a conventional personal computer or workstation, may be coupled (as indicated by the dashed arrow) to one or more of the devices in the sample separation apparatus 10 in order to receive information and/or control operation. For example, the control unit 70 may control the operation of the fluid drive unit 20 (e.g., set control parameters) and receive information therefrom regarding actual operating conditions (such as output pressure at the outlet of the pump, flow rate, etc.). The control unit 70 may also control the operation of the solvent supplier 25 (e.g. set the solvent or solvent mixture to be supplied) and/or the operation of the degasser 27 (e.g. set control parameters such as vacuum level, etc.) and may receive information from it regarding the actual working conditions (such as solvent composition supplied over time, flow rate, vacuum level, etc.). The control unit 70 may further control the operation of the sampling unit or injector 40 (e.g., control the synchronization of sample injection or sample injection with the operating conditions of the fluid drive unit 20). The sample separation unit 30 may also be controlled by the control unit 70 (e.g., selecting a particular flow path or column, setting an operating temperature, etc.), and in turn send information (e.g., operating conditions) to the control unit 70. Accordingly, the detector 50 may be controlled by the control unit 70 (e.g. with respect to spectral or wavelength settings, setting time constants, starting/stopping data acquisition) and send information (e.g. with respect to detected sample complexes) to the control unit 70. The control unit 70 may also control the operation of the fractionation unit 60 (e.g., in conjunction with data received from the detector 50) and provide data back.

Further, a heat-affected assembly 100 is arranged downstream of the injector 40 and upstream of the detector 50 in the sample separation apparatus 10. The heat-affected assembly 100 is configured to adjust the temperature of the mobile phase and the fluid sample, and independently or separately adjust the temperature of the sample separation unit 30. The heat-affected assembly 100 comprises a heat-affected zone, which here is constituted by a controllable first heat-affected cell 80 and an independently controllable second heat-affected cell 82. The control of each heat influencing unit 80, 82 may be performed by the control unit 70, the control unit 70 supplying respective and different control signals to the heat influencing units 80, 82. The first heat influencing unit 80 is configured to thermally influence the fluid sample and/or the mobile phase flowing through the conduit thermally conductively enclosed by the pre-processing assembly 90. The second heat influence unit 82 is configured to thermally influence a heat influence compartment 84 housing the sample separation unit 30. Therefore, the second heat-affected unit 82 will also control the temperature of the sample separation unit 30. The above-mentioned control unit 70 may be configured to control the heat influencing units 80, 82 to influence the heat on the one hand on the fluid sample and/or the mobile phase and on the other hand to influence the heat on the sample separation unit 30 separately from each other. Very advantageously, the heat influencing assembly 100 may thus be configured to influence the heat on the fluid sample and/or the mobile phase on the one hand and on the other hand to influence the heat on the sample separation unit 30 separately and differently, if desired. This introduces more degrees of freedom or design parameters that can be used to improve temperature regulation. For example, other target temperatures may be set for the fluid sample and the mobile phase as compared to the sample separation unit 30. In particular, the thermal influence on the fluid sample and/or the mobile phase may be performed by adjusting (e.g. regulating) the temperature of the fluid sample and/or the mobile phase. Independently thereof, the thermal influence on the sample separation unit 30 may be accomplished by adjusting (e.g., regulating) the temperature of the sample separation unit 30.

Additionally or alternatively, this additional degree of freedom may be used to emulate the performance on the sample separation apparatus 10 of a sample separation method developed for another sample separation apparatus (not shown in fig. 1), the sample separation apparatus 10 thereby functioning or behaving as the other sample separation apparatus performs the sample separation method. In other words, the thermal influence may be controlled to perform a simulation of a separation method of another sample separation apparatus (see reference numeral 110 in fig. 9) by the sample separation apparatus 10, such that the sample separation apparatus 10 behaves like the another sample separation apparatus 110 in terms of thermally influencing the fluid sample and/or the mobile phase and the sample separation unit 30.

It should be noted that in the illustrated embodiment, the control unit 70 for controlling the heat influencing units 80, 82 may be the same control unit 70 as described above which also controls the overall operation of the sample separation apparatus 10. In other embodiments, the control unit 70 for controlling the overall operation of the sample separation apparatus 10 may alternatively be another controller than the control unit 70 that controls the heat influencing units 80, 82 independently of each other.

A detailed configuration of a temperature adjustment assembly 100 according to an exemplary embodiment of the present invention, which temperature adjustment assembly 100 may be implemented in the sample separation apparatus 10 shown in fig. 1, will be explained with reference to fig. 2 to 9.

Fig. 2 is a schematic diagram of a heat-affected assembly 100 for sample isolation apparatus 10, wherein a first heat-affected cell 80 is integrated in or with selection valve 86 and a second heat-affected cell 82 is disposed inside heat-affected compartment 84, according to an exemplary embodiment.

As indicated by corresponding reference numerals in fig. 2, the heat-affected assembly 100 according to fig. 2 is arranged downstream of the injector 40 and upstream of the detector 50. The fluid flow direction is indicated by arrows in fig. 2. As shown, the heat-affected assembly 100 includes a heat-affected device composed of a first heat-affected unit 80 and a second heat-affected unit 82. The heat influencing means is configured to temper the fluid sample and/or the mobile phase and the sample separation unit 30. More specifically, the first heat influencing unit 80 heats (or cools) the fluid sample and/or the mobile phase as it flows through the first heat influencing unit 80. Independently thereof, the second heat-affected unit 82 heats (or cools) three parallel sample separation units 30 (which may be chromatographic separation columns) located in a heat-affected compartment 84 (such as a heating furnace or the like). Those skilled in the art will appreciate that the number of three parallel sample separation units 30 is merely an example, and other exemplary embodiments may use fewer (one or two) or more (four or more) parallel sample separation units 30. Thus, the number of parallel sample separation units 30 may be any number (and may for example be only two). Thus, the thermal pretreatment solvent and the sample can be controlled or adjusted independently of the thermal influence on the separation column. Thus, independent control of the temperature of the separation column and of the temperature of the mobile phase and the fluid sample is made possible. Under the control of the control unit 70, the first heat-influencing unit 80 may supply thermal energy to the mobile phase or the fluid sample (for heating) or may remove thermal energy from the mobile phase or the fluid sample (for cooling). Accordingly and also under the control of the control unit 70, the second heat-influencing unit 82 may supply thermal energy to the sample separation unit 30 (for heating) or may remove thermal energy from the sample separation unit 30 (for cooling). Thus, each of the heat influencing units 80, 82 may be configured as a heat source and/or a heat sink. Accordingly, each of the heat influencing units 80, 82 may comprise a heat exchanger thermally coupled to the fluid sample or mobile phase (in the case of the first heat influencing unit 80) or to the sample separation unit 30 (in the case of the second heat influencing unit 82). For example, each of the heat-influencing units 80, 82 may be a heating block or a cooling block.

The control unit 70, which may for example be a correspondingly programmed or programmable processor, may be configured to separately and individually control each of the heat influencing units 80, 82 to influence the heat on the fluid sample and/or the mobile phase or the sample separation unit 30, respectively, independently of each other. In particular, the first heat influencing unit 80 in combination with the control unit 70 may be configured to set a further target temperature or temperature distribution for the fluid sample and/or the mobile phase, compared to the target temperature or temperature distribution of the sample separation unit 30, which may be defined by the second heat influencing unit 82 in cooperation with the control unit 70. Thus, the control unit 70 may be configured to separately control the first and second heat influencing units 80, 82. For this reason, the control unit 70 may apply different control signals 71, 73 to the first heat influencing unit 80 with respect to the second heat influencing unit 82.

For example, any one of the first and second heat influencing units 80, 82 may be a block (such as a heating block or a cooling block, e.g., a heating plate or a cooling plate) that is heated or cooled, e.g., by a heating or cooling fluid (such as a hot or cold gas or liquid, etc.). It is also possible that in the case of ohmic heating, either of the first and second heat-influencing units 80, 82 may be heated by an electric current. When configured as a peltier element, the first and second heat-influencing units 80 and 82 may be selectively cooled or heated according to a flow direction of current applied to the peltier element. Thus, the heat-influencing elements 80, 82 may be configured to heat, cool, or selectively heat or cool the fluid sample and/or the mobile phase and/or the sample separation unit 30.

For example, the first heat-influencing unit 80 may be thermally decoupled from the second heat-influencing unit 82. This may facilitate independent control of the heat-influencing units 80, 82. Such a thermal decoupling may be achieved, for example, by a sufficient spatial distance between the first and second heat-influencing units 80, 82 and/or by arranging an insulating structure (not shown) between the first and second heat-influencing units 80, 82.

As shown, three sample separation units 30 (e.g., three different types of chromatographic separation columns) may be connected in parallel inside a heat-affected compartment 84 (e.g., a column oven, etc.). According to fig. 2, the first heat influencing unit 80 is arranged upstream of the second heat influencing unit 82. The heat-affected compartment 84 is for accommodating the second heat-affected cell 82 and the sample separation unit 30 therein. In other words, the second heat-affected cell 82 configured to thermally affect the sample separation unit 30 is disposed inside the heat-affected compartment 84.

Further, a first heat influencing unit 80 configured to thermally influence the fluid sample and/or the mobile phase is arranged upstream of the heat influencing compartment 84. As shown in fig. 2, the heat-affected assembly 100 includes a fluid selection valve 86, the fluid selection valve 86 being located upstream of the heat-affected compartment 84 and configured to select one of the sample separation units 30, for example, according to the requirements of a particular separation application. The mobile phase and/or fluid sample provided at the inlet of the selection valve 86 is directed to a selected one of the outlets of the selection valve 86 selected according to the switching state of the selection valve 86. In other words, depending on the switching position of the selection valve 86, the mobile phase and/or fluid sample flowing from the injector 40 may be directed into one of the three parallel fluid paths inside the heat affected compartment 84 to flow through a selected one of the three sample separation units 30. Advantageously, according to fig. 2, the first heat influencing unit 80 is integrated in or directly connected to a selector valve 86. Thus, column select valve 86 may be configured as a heating and/or cooling element for heating and/or cooling the mobile phase and/or fluid sample. This keeps the heat-affected assembly 100 compact and adjusts the temperature in the first and second heat-affected units 80, 82 spatially close together. Thus, according to fig. 2, artifacts caused by an undesired temperature balance of the mobile phase or fluid sample flowing through the conduit of the heat affected assembly 100 may be efficiently suppressed.

Fig. 3 is a schematic diagram of a heat-affected assembly 100 for a sample separation apparatus 10, wherein a first heat-affected cell 80 is disposed in a head portion of the sample separation cell 30 and a second heat-affected cell 82 is disposed inside the heat-affected compartment 84, according to an exemplary embodiment.

The embodiment of fig. 3 differs from the embodiment of fig. 2 in particular in that, according to fig. 3, a first heat influencing unit 80 and a second heat influencing unit 82 are arranged in a spatially overlapping manner. It is also possible that the second heat influencing unit 82 encapsulates or surrounds the first temperature regulating unit 80. According to fig. 3, both the first and second heat influencing units 80, 82 may be arranged inside the heat influencing compartment 84.

In this embodiment, a first heat influencing unit 80 configured to thermally influence the fluid sample and/or the mobile phase is thermally coupled to a head portion of the sample separation unit 30. The fluid sample and the mobile phase flow into the respective sample separation unit 30 at the head portion. In other words, the first heat influencing unit 80 heats or cools the mobile phase or fluid sample as it flows through the column head of the sample separation unit 30. It may be advantageous that the first heat-influencing unit 80 is arranged as close as possible to the column head in order to accurately control the sample temperature during separation. Therefore, the sample temperature is particularly critical at the head portion of the sample separation unit 30, since the actual separation process (absorption and desorption) takes place at this location.

Fig. 4 is a schematic diagram of a heat-affected assembly 100 for a sample separation apparatus 10, wherein a first heat-affected cell 80 is disposed between a selection valve 86 and a heat-affected compartment 84, and a second heat-affected cell 82 is disposed inside the heat-affected compartment 84, according to an exemplary embodiment.

The embodiment of fig. 4 differs from the embodiment of fig. 3 in particular in that according to fig. 4 a first heat influencing unit 80 is arranged downstream of a selector valve 86 and upstream of a heat influencing compartment 84. More specifically, the first heat-affected cell 80 may thermally affect the mobile phase and the fluid sample as the flow passes through the conduit connecting the selection valve 86 with the heat-affected compartment 84.

In the configuration according to fig. 4, the first and second heat influencing units 80, 82 are very close and close to the actual separation location of the fluid sample, while the independent controllability of the heat influencing units 80, 82 is further facilitated by their spatial separation.

Fig. 5 is a schematic diagram of a heat-affected assembly 100 for a sample separation apparatus 10, wherein a first heat-affected cell 80 is disposed upstream of a selection valve 86 and a second heat-affected cell 82 is disposed inside a heat-affected compartment 84, according to an exemplary embodiment.

The embodiment of fig. 5 differs from the embodiment of fig. 4 in particular in that, according to fig. 5, the first heat influencing unit 80 is arranged downstream of the injector 40 and upstream of the selection valve 86.

An advantage of this arrangement is that the first heat influencing unit 80 can be constructed in a highly compact manner since it acts on the mobile phase or the fluid sample before the flow path is divided into a plurality of parallel paths by the selector valve 86.

Fig. 6 is a schematic diagram of a heat-affected assembly 100 for a sample separation apparatus 10, wherein a first heat-affected cell 80 is disposed inside the heat-affected compartment 84, and a second heat-affected cell 82 is also disposed inside the heat-affected compartment 84, according to an exemplary embodiment. However, according to fig. 6, the heat-affected units 80, 82 are arranged in a non-overlapping manner.

In the embodiment of fig. 6, three pre-treatment assemblies 90 are provided for pre-heating the fluid sample and/or mobile phase. The pre-processing assembly 90 is housed in a parallel flow path inside the heat affected compartment 84. For each sample separation unit 30 and thus for each of the parallel flow paths selectable by the selection valve 86, an assigned pre-processing assembly 90 may be provided. Each pre-processing assembly 90 may tightly surround, in a thermally conductive manner, a respective capillary tube that carries a mobile phase or fluid sample within its interior. The pre-processing assembly 90 is heated or cooled by a first heat affected unit 80 under the control of the control unit 70, the first heat affected unit 80 also being disposed in the interior of the heat affected compartment 84. The pre-processing assembly 90 and the first heat-affected unit 80 are arranged upstream of the sample separation unit 30. The first heat influencing unit 80 is configured to thermally influence the fluid sample and the mobile phase and is for this purpose thermally coupled to the pre-processing assembly 90.

A second heat-affected cell 82 thermally coupled to the parallel arranged sample separation unit 30 is arranged downstream of the thermal pre-treatment assembly 90 and thus downstream of the first heat-affected cell 80, and is accommodated within a heat-affected compartment 84.

Fig. 7 is a schematic view of a heat influencing assembly 100 for a sample separation apparatus 10 according to an exemplary embodiment, wherein a first heat influencing unit 80 for thermally influencing a fluid sample and/or a mobile phase and a second heat influencing unit 82 for thermally influencing a fluid sample and/or a mobile phase and for thermally influencing a sample separation unit 30 are provided.

According to fig. 7, the fluid sample and/or the mobile phase may be tempered by a first heat influencing unit 80 and additionally and independently also by a second heat influencing unit 82. More specifically, the fluid sample and/or the mobile phase are arranged to be directly tempered by the first heat influence unit 80 (e.g., due to direct physical contact between the first heat influence unit 80 and a pre-processing assembly 90 surrounding a conduit through which the fluid sample and the mobile phase flow) and indirectly tempered by the second heat influence unit 82 (e.g., spaced apart by the first heat influence unit 80, as shown in fig. 7). For example, the fluid sample and/or the mobile phase may be heated by the second heat influencing unit 82 in terms of coarse temperature control, and may be selectively further heated or cooled by the first heat influencing unit 80 in terms of fine tuning of the temperature. In contrast, the sample separation unit 30, which may be arranged in the heat-affected compartment 84, may be tempered by the second heat-affected unit 82 only. Likewise, the control unit 70 may control the temperature conditioning functions of the first and second heat-influencing units 80, 82 independently or separately or individually.

In the embodiment of fig. 7, the first heat-influencing unit 80 may be a peltier element that may be operated by the control unit 70 to selectively heat or cool. Furthermore, the second heat influencing unit 82 may present an ohmically heatable mass, such as a heating block or the like.

As shown in fig. 7, the heat-affected compartment 84 may be directly tempered by the second heat-affected unit 82. For example, the heat-affected compartment 84, which may be present as a column oven, may be thermally coupled directly to the second heat-affected cell 82, e.g., may be mounted on a heating block.

The pre-processing assembly 90, through which the mobile phase and/or the fluid sample may flow, may be indirectly thermally coupled to the second heat influencing unit 82 (which may be in the form of a heating block). As shown, a first heat influencing unit 80 (particularly a peltier element) may be arranged sandwiched between a second heat influencing unit 82 and a pre-processing assembly 90. Thus, a majority of the thermal energy used to thermally influence the pre-processing assembly 90 may be provided by the second heat-influencing unit 82, while fine-tuning of the thermal influence of the pre-processing assembly 90 may be accomplished by the first heat-influencing unit 80. The latter may, for example, increase or decrease the temperature of the pre-processing assembly 90 by controlling the peltier elements accordingly. Thus, the described configuration and independent controllability of the heat-influencing units 80, 82 may allow for an efficient temperature control with a high flexibility.

FIG. 8 is a schematic view of a portion of a heat-affected assembly 100 in a heating compartment 84 of a sample separation device 10, wherein the sample separation unit 30 is heated by a convection mechanism 96 that operates only indirectly, according to an exemplary embodiment.

According to fig. 8, a parallel connected sample separation unit 30 (which may be present as a chromatographic separation column extending perpendicular to the paper surface of fig. 8) is accommodated inside a heat affected compartment 84. A circumferential air flow is generated outside the heat affected compartment 84 by a schematically illustrated convection mechanism 96. However, the resulting gas convection 94 only acts indirectly on the sample separation cell 30 to thermally influence it, without imposing the sample separation cell 30 to a direct gas flow. According to fig. 8, this is done by surrounding the sample separation unit 30 with a thermally conductive enclosure separating the sample separation unit 30 from the convective gas flow 94. The thermally conductive enclosure is comprised of a heat exchanger 92 and a flow shield 88.

Thus, the embodiment of fig. 8 shows a configuration of the second heat influencing unit 82 that enables heat influencing of the sample separation units 30 connected in parallel without the gas convection 94 acting directly on the sample separation units 30. In contrast to the gas convection 94 acting directly on the sample separation unit 30, the second heat influencing unit 82 is configured to thermally influence the sample separation unit 30 in case the gas convection 94 acts indirectly on the sample separation unit 30 according to fig. 8. This may be accomplished by providing a convection mechanism 96, the convection mechanism 96 for generating a gas convection 94 for thermal coupling with the sample separation cell 30, while the thermally conductive shielding structure 88 shields or spaces the gas convection 94 from the sample separation cell 30. Further, the thermally conductive shielding structure includes a heat exchanger 92, the heat exchanger 92 configured to facilitate heat exchange between the convective gas flow 94 and the sample isolation unit 30. The heat exchanger 92 may also be used to directly heat the sample separation unit 30. In addition, indirect convection, which is shielded with respect to the sample separation unit 30, may further facilitate proper heating of the sample separation unit 30. However, it has been found that when the sample separation unit 30 is prevented from being in direct contact with the convection flow, the performance of HPLC can be improved because it can suppress the formation of a significant temperature distribution between the inside and the outside of the cylindrical sample separation unit 30. Illustratively, such shielding may calm down the airflow around the sample separation unit 30, thereby improving separation performance.

As shown, a dividing wall of the heat affected compartment 84 (which may also be referred to as a column compartment) is provided as an outer housing. Reference numeral 92 denotes a heat exchanger, a heater, and a cooler of the system. Fig. 8 shows a cross section of a sample separation unit 30 (presented as an HPLC column). Reference numeral 88 denotes an air flow turning cowl or flow turning cowl. The arrows in fig. 8 illustrate the forced air flow or gas convection 94.

Advantageously, the shielding structure 88 may be mechanically coupled with a door (not shown) of the heat affected compartment 84 such that opening of such door by a user may automatically expose the sample separation unit 30 without separately disassembling the shielding structure 88. This ensures a user-friendly operation.

The embodiment of fig. 8 may or may not be combined with an independently controllable first heat influencing unit 80, e.g. implemented as described with reference to fig. 1 to 7.

FIG. 9 is a schematic diagram of a heat-affected assembly 100 of a sample separation apparatus 10, wherein operation of the heat-affected assembly 100 emulates the temperature-regulating behavior of another sample separation apparatus 110, according to an exemplary embodiment.

For example, the sample separation apparatus 10 may be constructed as described above with reference to fig. 1 and 2.

Another sample separation apparatus 110 may be configured to have a single common heat influencing device 199 inside the column oven 184. Through the column selection valve 186, one of three parallel fluid paths can be selected, each fluid path comprising a series connection of a pre-heater assembly 190 and the assigned chromatographic separation column 130. Heat influencing device 199 thermostats the fluid sample and mobile phase flowing through the respective pre-heater assemblies 190 and thermostats the sample separation unit 30. The sample separation device 110 may be configured to perform a chromatographic separation method that accomplishes a very specific separation task and is specifically configured according to the characteristics of the sample separation device 110. Such chromatographic methods may be stored in a database 99.

It may be desirable in certain circumstances to use other sample separation devices 10 to perform chromatographic separation methods developed specifically for the sample separation device 110. However, in view of the different characteristics of the sample separation devices 10, 110, performing a chromatographic separation method developed for the sample separation device 110 may yield another separation result (in particular another chromatogram) when performed on the sample separation apparatus 10.

By specifically configuring the sample separation apparatus 10 and in particular the heat affected assembly 100 thereof, performance of the chromatographic separation method may be backward compatible. Described in detail, by appropriately controlling the heat-affected units 80, 82 of the sample separation apparatus 10 by the control unit 70, it is possible to allow the configuration of the sample separation apparatus 10 so as to behave like the sample separation apparatus 110 in terms of temperature adjustment when performing a chromatographic separation method. In other words, with respect to preheating, adjusting the additional degrees of freedom of the heat-affected units 80, 82 separately or independently in the sample separation device 10 allows operating the sample separation device 10 to perform chromatographic separation methods developed for the sample separation device 110 to emulate the behavior of the sample separation device 110.

To this end, the control unit 70 may be configured to individually control each of the heat influencing units 80, 82 such that performing the separation method on the sample separation device 10 emulates the operation of another sample separation device 110 with respect to thermally influencing the fluid sample and/or the mobile phase and the sample separation unit 30. To control the heat influencing units 80, 82, the control unit 70 may determine and apply a transfer function describing the operation of the heat influencing units 80, 82 to behave like the heat influencing device 199 of the sample separation apparatus 110 in terms of temperature control. Thus, the control unit 70 may be configured to emulate the operation of the further sample separation device 110 based on said determined transfer function such that the sample separation device 10 behaves like the further sample separation device 110, in particular with respect to thermally influencing the fluid sample and/or the mobile phase and the sample separation unit 30, when performing the separation method (originally developed for the further sample separation device 110) on the sample separation device 10. The additional degrees of freedom or design parameters in the form of the independently controllable first heat-influencing unit 80 and the independently controllable second heat-influencing unit 82 can advantageously be used to provide the exemplary functions described.

Further advantageously, when performing the developed separation method on the sample separation device 10, emulating the temperature control behavior of the sample separation device 110 by controlling the sample separation device 10 accordingly may be synergistically combined with the time-dependent emulation of the solvent composition of the mobile phase of the sample separation device 110, in particular in terms of gradient runs. To this end, the target time-dependence of the solvent composition according to the chromatographic separation method developed for the sample separation device 110 may be transformed into a modified time-dependence (by correspondingly modifying the operation of the fluid driving unit 20 in combination with the solvent supply 25) such that the sample separation device 10 also behaves like the sample separation device 110 in terms of the time-dependence of the solvent composition of the mobile phase when performing the modified or adapted separation method. By taking this measure, the method transition from sample separation system 110 to sample separation system 10 can be made highly accurate.

Fig. 10 is a three-dimensional view of a first heat-affected cell 80 of a heat-affected assembly 100 of a sample separation apparatus 10 according to an exemplary embodiment. The illustrated first heat-affected cell 80 is configured as a metal-microfluidic (MMF) structure for heating or cooling a mobile phase and/or a fluid sample, and is provided integrated in a channel selection valve, such as the fluid selection valve 86 shown in fig. 2 or 9, or the like.

The heat affected unit 80 may include a plurality of metal structures connected by thermal bonding under high pressure and high temperature, and made of, for example, stainless steel foil. More specifically, the illustrated heat affected unit 80 is an interconnected annular structure 160 of metal foil, including an MMF heater 162 and an MMF cooler 164, and having a central through hole 166. Heating or cooling of the channel select valve 86 may be performed by the annular structure 160 acting as a pre-column liquid regulator.

Conventional column compartments require a solvent heater/cooler for each column, which affects the workload of manufacturing the instrument. These conventional devices are also located inside the compartment, affecting the temperature stability of the column environment.

In contrast to these conventional methods, the embodiment of fig. 10 uses MMF technology to embed the pre-column heater in the selector valve 86. The heat-affected cell 80 of fig. 10 can be obtained by using one or more plasma heaters (named from the manufacturing technology) and one or more peltier heaters in a sandwich structure packaged together with MMF (metal micro-fluidic). Thus, it may be possible to integrate the heat influencing function and the valve function in one more capable device, reducing the number of components required in the instrument for providing the function of the pre-column heater/cooler. Further, by replacing a plurality of (e.g., eight) column front heaters with one, a reduction in manufacturing effort can be achieved. The described embodiments also provide heating capability outside the column compartment. The liquid may be thermally affected before it enters the compartment interior, thereby avoiding internal condensation problems and temperature instability. Advantageously, the space in the compartment can be significantly reduced. Preferably, according to fig. 10, a sandwich structure may be formed, encapsulating the cooler and plasma heater in an MMF structure, most preferably in the head of the column select valve 86.

It should be noted that the term "comprising" does not exclude other elements, and the "a" or "an" does not exclude a plurality. Also elements described in association with different embodiments may be combined. It should also be noted that reference signs in the claims shall not be construed as limiting the scope of the claims.