DE102018114150A1 - Multi-dimensionally operable sample separator with common fluid drive - Google Patents

Abstract

A sample separation device (10) operable to separate at least two-dimensionally a fluidic sample, said sample separation device (10) establishing a fluid drive (20) for driving the fluidic sample in a first separation dimension and for driving the separated fluidic sample in a second separation dimension a first sample separator (30) for separating the fluid sample in the first separation dimension, means (32; 30, 70) for separating the fluid sample separated in the first separation dimension in the second separation dimension, and a detector (50) for detecting the having separate sample.

Description

TECHNICAL BACKGROUND

The present invention relates to a sample separation apparatus and a method for separating a fluidic sample.

In an HPLC, a liquid (mobile phase) is typically run at a very precisely controlled flow rate (for example in the range of microliters to milliliters per minute) and at a high pressure (typically 20 to 1000 bar and beyond, currently up to 2000 bar). , in which the compressibility of the liquid can be felt, through an adsorption medium, a so-called stationary phase (for example, in a chromatographic column), moved to separate individual components of a sample liquid introduced into the mobile phase from each other. Such an HPLC system is known, for example from the EP 0,309,596 B1 same Applicant, Agilent Technologies, Inc.

In the case of one-dimensional chromatography, it is possible for two or more different substances to be separated from the substances to be separated, moving at the same or approximately the same rate, and thus being found at the same or at approximately the same point in a chromatogram. For their further separation, two-dimensional chromatography can be used, in which in a second operation, other separation conditions such as another stationary phase and / or another mobile phase, or even just another state (for example, another temperature of the separation column), for renewed Chromatography of fractions from the first separation can be used, so that the difficult to distinguish after the first operation substances can now also be separated from each other. Orthogonality desired for two-dimensional separation can be accomplished by two sample separators or by using different mobile phases. In the latter case, a rinse between the two separations can be performed.

In two-dimensional liquid chromatography, a run in a second dimension (corresponding to a second separation step) can be started after a fraction in a sample section of the fluidic sample to be separated has been detected in the first dimension (corresponding to a first separation step) or recognized in the approach. If another fraction in another sample section of the fluidic sample to be separated is detected in the first separation stage, while a preceding fraction in the second separation stage is still being analyzed, this further fraction can be temporarily stored. A conventional approach is to buffer the volume of the fraction (ie the associated sample separation portion) in one of several liquid containers, such as capillaries between fluid ports of a fluidic valve, where the sample volume associated with, for example, another fraction, by switching the flow path to another valve position such capillary is led to the intermediate storage. Typically, two capillaries associated with the fluidic valve are used for caching to maintain caching uninterrupted, i. without loss of continuously pumped sample liquid to allow. Once the analysis has been made in the second dimension, the capillary with the next cached sample portion (eg, with a next fraction) is introduced into the second dimension flow path and a new separation in the second dimension is started.

Two-dimensional sample separation devices often have a high complexity.

EPIPHANY

It is an object of the invention to make a multi-dimensionally operable sample separation device compact. The object is achieved by means of the independent claims. Further embodiments are shown in the dependent claims.

In accordance with an exemplary embodiment of the present invention, there is provided a sample separation apparatus operable to separate at least two-dimensionally a fluidic sample, the sample separation apparatus comprising a fluid drive capable of driving the fluidic sample in a first separation dimension and driving the separated fluidic sample in a second separation dimension a first sample separator for separating the fluidic sample in the first separation dimension, means for separating the separated in the first separation dimension fluidic sample in the second separation dimension and a detector for detecting the separated sample.

According to another exemplary embodiment of the present invention, a method for separating a fluidic sample by means of a sample separation device (in particular having the features described above) is provided for separating the fluidic sample in a first separation dimension and for separating the separated fluidic sample in a second separation dimension wherein the method comprises operating a common fluid drive to drive the mobile phase fluid sample into separating the fluidic sample by means of a first sample separator in the first separation dimension, separating the fluidic sample separated in the first separation dimension (in particular by means of a second sample separator) in the second separation dimension, and detecting the separated one fluidic sample by means of a detector.

In the context of this description, the term "fluidic sample" is understood in particular to mean a liquid and / or gaseous sample, optionally with solid constituents.

According to an exemplary embodiment of the invention, there is provided a sample separation apparatus comprising two-dimensionally separating a fluidic sample into fractions and then into sub-fractions using a common fluid drive to drive a mobile phase and the fluidic sample therein. By skillful switching fluidic valves or other switching elements (for example, fluidic restrictions), the fluidic sample first in a first sample separator according to a first separation dimension and subsequently in a second device (for example, another second sample separator) are separated according to a second separation dimension, For both separation dimensions, the same fluid drive can be used to drive the fluidic sample and the mobile phase. This sharing of only one fluid drive for the promotion of mobile phases and the fluidic sample in both (or all) separation dimensions results in a compact sample separation device with two- or multi-dimensional separation functionality. Conventional two-dimensional sample separation devices typically implement a separate fluid drive per separation dimension, i. a total of several fluid drives.

In addition, additional embodiments of the sample separation device and the method will be described.

According to one embodiment, the device may comprise a second sample separation device (in particular a second chromatographic separation column, which may be different from the first chromatographic separation column, in particular another stationary phase) for separating the separated fluidic sample in the second separation dimension. In a corresponding manner, the separation of the separated fluidic sample in the second separation dimension can be effected by means of a second sample separation device. The effluent of the separated fluidic sample from the first separation dimension may then flow into the second chromatographic separation column or the like. Alternatively or additionally, the sample separation device may comprise a control device which is operated to separate the separated fluidic sample in the first sample separation device with a different mobile phase and / or at a different temperature than in the separation of the fluidic sample in the first separation dimension. Correspondingly, the separation in the second separation dimension can take place by means of another mobile phase or at a different temperature than during the separation of the fluidic sample in the first separation dimension. For example, in the last described embodiment, the first sample separator (i.e., in particular a first chromatographic separation column) may also be used a second time to separate the already separated fluid sample by returning the effluent from the first sample separator to an inlet of the first sample separator. Then, for example, using a different mobile phase compared to the first separation dimension in the second separation dimension by the different solvent composition of the mobile phase, a continued separation of the already pre-separated fluidic sample can be carried out.

According to one embodiment, the sample separation device can only have exactly the one fluid drive, in particular designed as the only high-pressure pump of the sample separation device. Such a high-pressure pump can promote mobile phase and fluidic sample, for example, with a pressure of at least 600 bar, in particular at least 1200 bar, more particularly at least 1800 bar. Such a fluid drive can be formed, for example, from one or more piston-cylinder pairs, which can be connected in fluidic parallel and / or fluidic series, for example, and can be driven coordinated to convey fluid at the most constant flow rate possible.

According to one embodiment, the sample separation device can only have exactly one detector, in particular designed as a single fluorescence detector of the sample separation device or as a detector of another type. By using only a single detector, the compactness of the sample separator can be further reduced. For example, the detector may detect outflow from the second separation dimension. However, it is also alternatively or additionally possible to switch the detector such that it detects the result of this first separation after the outflow of the fluidic sample from the first separation dimension. A fluorescence detector can detect fluorescent light from separate fractions of the fluidic sample with which they are intrinsically provided or specially labeled.

According to one embodiment, the detector may be designed as a destructive detector. Frequently, detectors are divided into destructive and non-destructive devices. In the former, the analyte is consumed for detection (for example, in a chemical reaction). In this context, a destructive detector can therefore be understood as meaning in particular a detector which destroys the separate and to be detected fluidic sample during the detection process. An example of this is a mass spectrometer.

It is also possible to use several detectors.

According to one embodiment, the sample separation device may be formed as a modular arrangement of a plurality of modules, which can be arranged as a stack, in particular as exactly one stack. By allowing a user to freely assemble and combine the individual modules of the sample separation device (eg, fluid drive, respective sample separator, detector, injector, fractionator, switching unit, etc.), the flexibility in operating the sample separation device is high. Due to the high compactness of the three-dimensionally operable sample separation device, it is possible to arrange the required modules on a single stack. This allows a space-saving arrangement of the sample separation in a laboratory.

According to one exemplary embodiment, the sample separating device may be configured such that, after separating a first section of the fluidic sample in the first separation dimension, it is temporarily stored in a first sample receiving volume, after a subsequent separation of a second section of the fluidic sample in the first separating dimension being temporarily stored in a second sample receiving volume and thereafter, the first portion and the second portion are successively subjected to the separation in the second separation dimension from the respective first and second sample receiving volumes, respectively. This is for example in 10 to 13 shown.

According to one embodiment, the sample separation device may be configured to perform the separation in the first separation dimension and the separation in the second separation dimension alternately, in particular only alternately. It is thus possible to operate the sample separation device in such a way that a first-dimensional or a second-dimensional separation is performed alternately. An alternating instead of parallel operation enables a compact construction of the sample separation device.

According to one embodiment, the sample separation device may comprise a first fluid valve with fluid connections, one of which is connected to the fluid drive, another to the first sample separation device, another to the second sample separation device and yet another to the detector. The first fluid valve may be, for example, a rotor valve. Such a rotor valve may comprise a stator and a rotor, which are rotatable relative to each other. The plurality of fluid connections (ports) can be provided on the stator, to which a respective fluidic module or fluidic component can be connected via a capillary line or the like. In addition, fluid lines (for example grooves) can be formed on the rotor and / or on the stator, which can fluidically couple the individual fluid connections to one another or can fluidically decouple from one another by rotating between the rotor and the stator. The different fluid ports may fluidly couple or fluidly decouple the first sample separator, the second sample separator and the detector to perform the described different modes of operation.

According to one embodiment, the sample separator may include a sample injector connected to still another of the fluid ports of the first fluid valve. A fluidic sample can be injected into the sample separation device via the sample injector, in particular into a separation path between the fluid drive and the respective sample separation device for the first separation dimension or for the second separation dimension. The injector may be connected to a further fluid port of the first fluid valve. As a result, the first fluid valve can operate a large number of fluidic components in a compact manner and bring them into operative connection with one another.

In one embodiment, the sample separation device may include a second fluid valve connected to one or more of the fluid ports of the first fluid valve. The second fluid valve may in particular be connected between two fluid ports of the first fluid valve.

According to one embodiment, the sample separation device can have at least one intermediate storage volume, which is designed for the temporary buffering of a fluid portion, in particular of a fluidic sample. In particular, the at least one intermediate storage volume can be connected between fluid connections of at least one intermediate storage valve, in particular of two intermediate storage valves, wherein the at least one intermediate storage valve can be connected to the second fluid valve. The buffer volumes may, for example, be sample loops formed between fluid ports of or of the respective buffer valve. In the figures, a vividly described as a parking deck arrangement of many Buffer volumes, each shown between fluid ports of one of two intermediate storage valves (see for example 1 ). In this way, separate fluid portions may be temporarily stored in the first (or nth) separation dimension and subsequently transferred for separation into the second (or (n + 1) th) separation dimension.

In one embodiment, in the two-dimensional sample separation in a first fluid valve switching state, the fluidic sample may be feasible through a first of the at least one latch volume and in another fluid valve switching state be feasible through a second of the at least one latch volume. In the two-dimensional sample separation operation, the buffer volumes can thus be used for temporarily storing fluid portions which have been separated in the first separation dimension.

According to one embodiment, after separation in the second separation dimension, the method may comprise returning the fluidic sample separated in the second separation dimension (in particular, returning an outflow of the detector) to the first sample separation device for separating the already twice separated sample in a third separation dimension. If, after separation in the second separation dimension, the outflow of the second separation dimension is returned to the first sample separation device, three-dimensional sample separation is also possible. The sample separation device according to an exemplary embodiment is thus very flexible for all single- or multi-dimensional separation processes can be used and yet compact.

According to one embodiment, after separation in the third separation dimension, the method may comprise feeding the already three times separated sample to the second sample separation device to separate the already three times separated sample in a fourth separation dimension. Thus, a separation in a fourth separation dimension is also possible if the sample flowing out of the third separation dimension is returned to the second sample separation device. There, the separation of the fourth separation dimension is performed and has already been carried out the separation in the second separation dimension.

The sample separation device can be a microfluidic measuring device, a liquid chromatography device or an HPLC or UHPLC system. However, other applications are possible.

The sample separation device may have in common for all separation stages only one pump for moving the respective mobile phase. Such a pump may, for example, be adapted to convey the mobile phase through the system at a high pressure, for example from a few hundred bars up to 1000 bars or more.

Alternatively or additionally, the sample separation apparatus may include a sample fractionator for fractionating the separated components. Such a fractionator may carry the various components, for example, into different liquid containers. The analyzed fluid sample can also be discarded.

Preferably, the sample separation device in one or each of the separation stages or for all separation stages together have a detector for the detection of the separated components. Such a detector may generate a signal which can be observed and / or recorded and which is indicative of the presence and amount of sample components in the fluid flowing through the system.

list of figures

• 1 bis 4 zeigen unterschiedliche Betriebsmodi eines Probentrenngeräts gemäß einem exemplarischen Ausführungsbeispiel der Erfindung.
• 5 zeigt einen einzigen Stapel aus Modulen, die ein Probentrenngerät gemäß einem exemplarischen Ausführungsbeispiel der Erfindung bilden.
• 6 bis 9 zeigen unterschiedliche Betriebsmodi eines Probentrenngeräts gemäß einem anderen exemplarischen Ausführungsbeispiel der Erfindung.
• 10 bis 13 zeigen unterschiedliche Betriebsmodi eines Probentrenngeräts gemäß einem anderen exemplarischen Ausführungsbeispiel der Erfindung, das dem Prinzip gemäß 1 bis 4 entspricht.
• 14 bis 17 zeigen unterschiedliche Betriebsmodi eines Probentrenngeräts gemäß einem anderen exemplarischen Ausführungsbeispiel der Erfindung, das ähnlich dem Prinzip gemäß 6 bis 9 ist.
• 18 und 19 zeigen unterschiedliche Betriebsmodi eines Probentrenngeräts gemäß einem anderen exemplarischen Ausführungsbeispiel der Erfindung, das mit besonders geringem Aufwand ausgebildet werden kann.
• 20 zeigt ein Probentrenngerät gemäß einem anderen exemplarischen Ausführungsbeispiel der Erfindung als Alternative zu jenem gemäß 18 und 19.
• 21 zeigt ein Probentrenngerät gemäß einem anderen exemplarischen Ausführungsbeispiel der Erfindung, das mit geringem Aufwand ausgebildet werden kann.
• 22 und 23 zeigen unterschiedliche Betriebsmodi eines Probentrenngeräts gemäß einem anderen exemplarischen Ausführungsbeispiel der Erfindung.
• 24 und 25 zeigen unterschiedliche Betriebsmodi eines Probentrenngeräts gemäß noch einem anderen exemplarischen Ausführungsbeispiel der Erfindung mit einer gemeinsamen Probentrenneinrichtung für beide Trenndimensionen.
• 26 bis 32 zeigen unterschiedliche Betriebsmodi von Probentrenngeräten gemäß exemplarischen Ausführungsbeispielen der Erfindung, die auch ein Fluidventil in Pump-Purge-Position zeigen.

Other objects and many of the attendant advantages of embodiments of the present invention will be readily appreciated and become better understood by reference to the following more particular description of embodiments taken in conjunction with the accompanying drawings. Features that are substantially or functionally the same or similar are given the same reference numerals.

• 1 to 4 show different modes of operation of a sample separator according to an exemplary embodiment of the invention.
• 5 shows a single stack of modules forming a sample separation apparatus according to an exemplary embodiment of the invention.
• 6 to 9 show different operating modes of a sample separation device according to another exemplary embodiment of the invention.
• 10 to 13 show different operating modes of a sample separation device according to another exemplary embodiment of the invention, which according to the principle 1 to 4 equivalent.
• 14 to 17 show different operating modes of a sample separator according to another exemplary embodiment of the invention, which is similar to the principle according to FIG 6 to 9 is.
• 18 and 19 show different modes of operation of a sample separation device according to another exemplary embodiment of the invention, which can be formed with very little effort.
• 20 shows a sample separation device according to another exemplary embodiment of the invention as an alternative to that according to FIG 18 and 19 ,
• 21 shows a sample separation device according to another exemplary embodiment of the invention, which can be formed with little effort.
• 22 and 23 show different operating modes of a sample separation device according to another exemplary embodiment of the invention.
• 24 and 25 show different modes of operation of a sample separation apparatus according to yet another exemplary embodiment of the invention with a common sample separation device for both separation dimensions.
• 26 to 32 show different operating modes of sample separation devices according to exemplary embodiments of the invention, which also show a fluid valve in pump purge position.

The illustration in the drawing is schematic.

Before the figures are described in detail, some more general considerations of the present inventors of exemplary embodiments of the invention are presented.

According to an exemplary embodiment of the invention, a sample separation device is provided which, for example, can be designed as a single stack of modules and permits a multi-dimensional sample separation with a common fluid drive.

Thus, according to an exemplary embodiment, it is possible to use a single pump and a detector (possibly a single detector) while still performing two-dimensional liquid chromatography (2D-LC). For example, the detector may be a destructive detector for detection after the first separation dimension and without using a splitter. By allowing a single stack instead of two stacks for the design of the two-dimensionally operable sample separator, a compact configuration is provided. The flexible operation of the sample separation device in a compact manner with only one fluid drive (and possibly only one detector) can be achieved by the combination of, for example, four fluid valves (namely, the first and second fluid valves described above, and preferably two further intermediate storage valves). The detector may be used alternately for both separation dimensions and may be a destructive detector. Such a sample separation device can also be advantageously used for a heart-cutting operating state. It is possible to connect a separate detector with a splitter.

The embodiment described makes it possible to carry out a further analysis, which can be repeated as often as desired, in a respective new separation dimension with the outflow from a preceding separation dimension. As a result, an unlimited number of liquid chromatography separation dimensions can be implemented.

According to an exemplary embodiment, a one-dimensional sample separator of pump, sample injector, sample separator, and sequential two-dimensional sample separation detector may be configured, for example merely by adding fluid valves and adjusting a control of the sample separator. Clearly, in a first separation dimension, a fractionation of a fluidic sample can take place and the fractionation result can be fed to a second separation dimension in order to further separate the sample. In particular, in an exemplary embodiment of the invention, separation in the first dimension and in the second dimension is not simultaneous, but sequentially. The preferably single pump therefore promotes clearly at any time either in a first dimension or in a second dimension.

For example, according to one exemplary embodiment of the invention, first-dimension separation may occur until all of the sample receiving volumes of an associated sample receiving valve are filled with sample portions. Now the operation can be stopped or interrupted, so that no flow from the first dimension out is more possible. It is now possible to carry out a processing of the already separated fluidic sample in the second separation dimension, whereby the sample receiving volumes of the associated sample receiving valve are emptied. As soon as a corresponding execution is completed, an outflow from the first sample separation dimension can be reactivated.

1 to 4 show different operating modes of a designed as a HPLC device sample separator 10 according to an exemplary embodiment of the invention. In 1 is the sample separator 10 in a first position of shown below in more detail fluidic valves belonging to a first separation dimension of a two-dimensional sample separation. In 2 is the sample separator 10 according to 1 shown in a second position of the valves, also belonging to the first separation dimension of the two-dimensional sample separation. 3 shows a first switching state of the fluid valves associated with a second separation dimension of the two-dimensional sample separation. In 4 a second position of the valves is shown in the context of the second separation dimension of the two-dimensional sample separation. The respective allowed river paths are 1 to 4 refer to.

This in 1 Sample separator shown 10 is operable to two-dimensionally separate a fluidic sample. The sample separator 10 is formed, the operating mode with the separation in the first dimension ( 1 . 2 ) and the other operating mode with the separation in the second dimension ( 3 . 4 ) alternately.

For this purpose, the sample separation device 10 exactly one designed as a high-pressure pump fluid drive 20 configured to drive the fluidic sample in a first separation dimension and to drive the separated fluidic sample in a second separation dimension. The sample separator 10 only has exactly the one fluid drive 20 on, therefore, as the only high pressure pump of the sample separator 10 acts. For example, the fluid drive 20 be designed as a piston pump or serial and / or parallel arrangement of piston pumps and serves both separation dimensions.

In addition, the sample separator shows 10 a first sample separation device 30 in the form of a first chromatographic separation column for separating the fluidic sample in the first separation dimension. Furthermore, the sample separator contains 10 a second sample separator 32 for separating the separated fluidic sample in the second separation dimension.

In addition, a detector 50 (For example, a quadrupole time-of-flight (Q-TOF) detector or a fluorescence detector with a flux line) for detecting the separated sample. In the illustrated embodiment of the sample separation device 10 is just the one detector 50 intended. In other embodiments, a supplemental detector may be provided.

Furthermore, the sample separator 10 a first fluid valve 84 with fluid connections 86 one of which is attached to the fluid drive 20 , another to the first sample separator 30 , another to the second sample separation device 32 and another to the detector 50 are connected.

In addition, in the sample separator 10 a sample injector 40 or autosampler for injecting a fluidic sample to be separated provided to yet another of the fluid ports of the first fluid valve 84 connected. A second fluid valve 88 is at two of the fluid ports of the first fluid valve 84 connected.

Furthermore, the sample separation device 10 a sample fractionator (not shown in U.S. Pat 1 to 4 ) for fractionating the separated fractions.

In addition, according to 1 to 4 Between storage volumes 96 implemented for temporary caching a fluid portion of fluidic sample. The buffer volumes 96 are between fluid connections 86 a respective one of two intermediate storage valves 94 connected. In addition, the two intermediate storage valves 94 to the second fluid valve 88 connected. The two intermediate storage valves 94 connected to the second fluid valve 88 are connected, have a plurality of buffer volumes 96 in the form of capillary sections between two respective fluid ports 86 the intermediate storage valves 94 , They serve as parking decks, that is to say for the temporary storage of sample sections which have been separated in the first separation dimension. In two-dimensional sample separation in a first fluid valve switching state, the fluidic sample is through a first of the buffer volumes 96 feasible. In another fluid valve switching state, the fluidic sample is through a second of the buffer volumes 96 feasible.

After separation in the second separation dimension can be a return of the already twice separated fluidic sample to the first sample separation device 30 to separate the already twice separated sample in a third separation dimension. After separation in the third separation dimension, it is again possible to feed the sample, which has already been separated three times, to the second sample separation device 32 be carried out to separate the already three times separated sample in a fourth separation dimension. In this way, with the sample separator 10 also a three-dimensional, four-dimensional, etc. (generally n-dimensional) sample separation can be performed.

Referring to 1 to 4 So it's a sample separator 10 according to an exemplary embodiment of the invention, the only a single fluid drive 20 needed. This allows multiple heart cutting and High-resolution two-dimensional liquid chromatography separation analyzes are performed. In addition, there is only one detector 50 which can be used both for the analysis in the first separation dimension and for the analysis in the second separation dimension. Given the implementation of only a single high-pressure pump as a fluid drive 20 becomes the first-dimensional analysis ( 1 . 2 ) and the second-dimensional analysis ( 3 . 4 ) are not performed in parallel, but successively. This may be advantageously assisted by providing two additional base positions of the valve setup, besides the setups for temporarily caching or parking sample sections (each corresponding to a chromatographic peak) in the buffer volumes 96 the intermediate storage valves 94 , So it's with a single detector 50 (In particular, a destructive detector) an analysis possible without an additional split is implemented. In the periods in which the peaks are cut out of the first-dimensional analysis, no first-dimensional separation data are determined. However, high sensitivity and full flow rate are ensured in both separation dimensions.

1 to 4 also show a control device 70 for controlling the fluid drive 20 or even the entire sample separation device 10 , The control device 70 may be used to separate the fluidic sample separated in the first separation dimension in the second separation dimension by means of the first sample separation device 30 be formed with a different mobile phase than in the separation of the fluidic sample in the first separation dimension. A desirable orthogonality between the two separation dimensions is then not achieved by two different sample separators 30 . 32 but accomplished by different solvent compositions, which by means of the first sample separation device 30 perform the separations. The sample separator 32 can be omitted in such an embodiment.

5 shows a single stack 82 from modules 80 containing a sample separator 10 form according to an exemplary embodiment of the invention. 5 shows the high compactness and simplicity of the modular sample separation device 10 , which can be used for two-dimensional sample separation. The sample separator 10 according to 1 to 4 For example, as a modular arrangement of a plurality of modules 80 (according to reference numerals 20 . 30 . 40 . 50 . 70 , etc.), which are preferably exactly one stack 82 can be arranged (see 5 ), more precisely stackable on each other.

6 to 9 show different operating modes of a sample separator 10 according to another exemplary embodiment of the invention. In 6 is the sample separator 10 shown in a first position of the fluidic valves, associated with a first separation dimension of a two-dimensional sample separation. In 7 is the sample separator 10 according to 6 shown in a second position of the valves, also belonging to the first separation dimension of the two-dimensional sample separation. 8th shows a first switching state of the fluid valves associated with a second separation dimension of the two-dimensional sample separation. In 9 a second position of the valves is shown in the context of the second separation dimension of the two-dimensional sample separation. The respective allowed river paths are 6 to 9 refer to.

The injector 40 in turn can be configured as an autosampler. A first detector 50 may be a Q-TOF detector, whereas a second detector 51 may be a DAD (diode array detector) detector. Again, the intermediate storage valves 94 designed as parking decks. The embodiment according to 6 to 9 also uses only a common high-pressure pump as a single fluid drive 20 for driving fluidic sample and mobile phase. The first-dimensional analysis ( 6 . 7 ) and the second-dimensional analysis ( 8th . 9 ) are performed one after the other. This corresponds to two additional base positions of the valve setup, apart from the further setups for parking out the sample sections corresponding to the individual peaks from the first separation dimension in the intermediate storage volumes 96 the intermediate storage valves 94 (clearly referred to as parking decks).

In the illustrated embodiment, a non-destructive one-dimensional detector 51 be implemented, with the parking decks behind the detector 51 can be arranged. The first-dimensional data is continuously recorded without losing data. An additional destructive detector 50 , such as a mass spectrometer, can be placed in front of the detector 51 be connected via a split. Multiple parking positions can be used multiple times in succession, and a virtually unlimited number of liquid chromatography separation dimensions can be implemented.

10 to 13 show different operating modes of a sample separator 10 according to another exemplary embodiment of the invention, which according to the principle 1 to 4 equivalent.

In addition or alternatively to the components described above has the Sample separation device 10 according to 10 to 13 a first sample loop 71 for receiving a given volume of fluidic sample and a second sample loop 73 to receive a given volume fluidic sample and a first fractionator 75 for fractionating the separated fluidic sample and a second fractionator 77 for fractionating the separated fluidic sample. A respective trial loop 71 . 73 can also be through a buffer valve 94 with buffer volumes 96 replaced or implemented. In the fractionators 75 . 77 For example, fractions of the fluidic sample can be taken up in separate sample containers, fed to separation columns, etc.

The operating mode according to 10 corresponds to a separation in the first separation dimension. The result of this first separation may be in the first sample loop 71 be cached or parked.

The operating mode according to 11 also corresponds to a separation in the first separation dimension. The result of this first separation may be in the second sample loop 73 be cached or parked.

After going through the operating modes according to 10 and 11 are both sample loops 71 . 73 filled with separated in the first separation dimension fluidic sample.

According to 12 will be the one in the first sample loop 71 cached fluidic sample in the second sample separation device 32 and from there into the detector 50 promoted, whereby the separation in the second separation dimension is accomplished.

According to 13 will be in the second sample loop 73 cached fluidic sample in the second sample separation device 32 and from there into the detector 50 promoted, whereby the separation in the second separation dimension is accomplished.

14 to 17 show different operating modes of a sample separator 10 according to another exemplary embodiment of the invention, which according to the principle 6 to 9 similar.

In contrast to 10 to 13 is according to 14 to 17 the detector 50 upstream of the (also serving as fractionators or with fractionators 75 . 77 cooperating) sample loops 71 respectively. 73 arranged. In other words, the fluidic sample first flows into the detector 50 and only then into the respective sample loop 71 respectively. 73 , This can be done in the detector 50 be recognized, what later in the respective test loop 71 respectively. 73 is filled. This possibly an improved control. The effluent of the second separation dimension can be fractionated.

The operating mode according to 14 corresponds to that according to 10 , and the operating mode according to 15 corresponds to that according to 11 , However, according to 16 the second sample loop 73 with the result of the second sample separation of the fluidic sample from the first sample loop 71 filled. According to 17 then becomes the fluidic sample, which has already been separated twice and in the second sample loop 73 contained in a third separation dimension of the second sample separator 32 further separately. By repeating this procedure, n-dimensional separation (n = 3, 4, 5, ...) is possible.

18 and 19 show different operating modes of a sample separator 10 according to another exemplary embodiment of the invention, which can be formed with very little effort.

According to 18 and 19 only a single sample separator 30 is implemented. However, in this embodiment, it is possible to achieve two-dimensional orthogonality by using different mobile phases for a first separation and for a second separation.

18 shows the separation in the first dimension. Upon completion of this separation, the separate fluidic sample becomes in a sample receiving volume 96 of the sample receiving valve 94 recorded and cached there. 19 shows the separation in the second dimension. For this purpose, the separate fluidic sample from the sample receiving volume 96 of the sample receiving valve 94 again through the single sample separator 30 led and separated for the second time.

The sample separator 30 can also be replaced by an array of parallel and selectable separation columns. The sampler or sample injector 40 can the fluid drive 20 be immediately downstream.

20 shows a sample separator 10 according to another exemplary embodiment of the invention as an alternative to that according to FIG 18 and 19 ,

In contrast to 18 and 19 is according to 20 the sample injector 40 upstream of the first fluid valve 84 arranged. Furthermore, according to 20 a bridge 81 between two fluid ports of the fluid valve 84 provided as additional parking space for temporary storage of fluidic sample can serve.

21 shows a sample separator 10 according to another exemplary embodiment of the invention, which can be formed with little effort.

According to 21 are two fluidic shorts 83 . 85 implemented. As a result, a Vorschaltventil can be saved.

Each sample receiving valve 94 has a fluidic short circuit 83 . 85 , In accordance with 21 upper sample receiving valve 94 this is the starting position, according to 21 lower sample intake valve 94 the end position. The pump pumps over the fluidic short circuit 83 into the column, according to 21 lower sample intake valve 94 injected until the fluidic short circuit 83 is full. Is the last position according to 21 lower sample intake valve 94 (fluidic short circuit 85 ), the fluid valve switches 84 around, and according to 21 upper sample receiving valve 94 can record further sample sections.

Third and further separation dimensions are possible according to this embodiment, if parking is provided for this purpose (clearly, no switching of the fluid valve takes place 84 during a run, reducing the capacity of the sample volumes 96 a sample receiving valve 94 minus a respective sample receiving volume 96 per dimension is usable).

The sample separator 30 can also be replaced by an array of parallel and selectable separation columns. The sampler or sample injector 40 is from the pump or the fluid drive 20 separated by the fluid valve 84. Alternatively, the sampler of the pump can be immediately downstream.

22 and 23 show different operating modes of a sample separator 10 according to another exemplary embodiment of the invention.

According to this embodiment, there is an additional fluid control valve or first fluid valve 84 which graphically illustrates a configuration according to a separation in a first separation dimension (cf. 22 ) and another configuration corresponding to a separation in a second separation dimension (cf. 23 ) switches. The fluid valve 84 on the other hand, switches to a respective fluid receiving volume 96 a respective one of the two fluid receiving valves 94 for caching a respective packet of separate fluidic sample. With the illustrated configuration, sequential execution of the separation in the first separation dimension and in the second separation dimension is enabled.

With advantage this can be done in 22 and 23 illustrated embodiment with only one intermediate storage valve 94 be formed. In particular, an operation with three or more separation dimensions is possible. Furthermore, such an embodiment can be made using modules known per se 80 be formed by such modules 80 be combined accordingly. In addition, the configuration shown, starting from a one-dimensional sample separation device ago up or retrofitted formable.

24 and 25 show different operating modes of a sample separator 10 according to another exemplary embodiment of the invention with a common sample separator 30 for both separation dimensions.

The embodiment according to 24 and 25 offers similar advantages as the embodiment according to 22 and 23 but has only a single sample separator 30 on. 24 shows a separation in the first separation dimension, whereas 25 shows a separation in the second separation dimension.

A control valve or first fluid valve designed as a 6-position 7-port valve 84 also solves potential problems associated with the occurrence of two shortcuts (compare reference numbers) 83 . 85 ).

26 to 32 show different operating modes of sample separation equipment 10 according to exemplary embodiments of the invention, which also includes the first fluid valve 84 in pump-purge position. These figures also show a waste connection 60 as well as a control block 91 for controlling an operation with a separation in a first separation dimension and a separation in a second separation dimension, respectively. This control block 91 in particular, the components according to reference numerals 88 . 94 contain.

For a control valve or first fluid valve designed as a 6-position 7-port valve 84 shows 26 a separation in a first separation dimension, 27 a separation in a second separation dimension and 28 a pump-purge operation.

For a control valve or first fluid valve designed as an 8-position 9-port valve 84 shows 29 a separation in a first separation dimension, 30 a separation in a second Separation dimension 31 a detector and sample loop rinse operation and 32 a waste operation.

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

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

Cited patent literature

• EP 0309596 B1 [0002]

Claims (20)

A sample separation device (10) operable to separate at least two-dimensionally a fluidic sample, the sample separation device (10) comprising: a fluid actuator (20) adapted to drive the fluidic sample in a first separation dimension and to drive the separated fluidic sample in a second separation dimension; a first sample separator (30) for separating the fluid sample in the first separation dimension; means (32, 30, 70) for separating the fluidic sample separated in the first separation dimension in the second separation dimension; a detector (50) for detecting the separated sample. Sample separating device (10) according to Claim 1 , having only exactly one fluid drive (20), which is designed in particular as the only high-pressure pump of the sample separation device (10). Sample separating device (10) according to Claim 1 or 2 comprising only one detector (50), especially designed as a single fluorescence detector (50) of the sample separation device (10). Sample separating device (10) according to one of Claims 1 to 3 , wherein the detector (50) is designed as a destructive detector (50). Sample separating device (10) according to one of Claims 1 to 4 formed as a modular arrangement of a plurality of modules (80), which can be arranged as a stack (82), in particular as exactly one stack (82). Sample separating device (10) according to one of Claims 1 to 5 wherein the sample separation device (10) is designed, after separation in the second separation dimension, to carry out a return of the fluid sample separated in the second separation dimension to the first sample separation device (30) in order to separate the already twice separated sample in a third separation dimension. Sample separating device (10) according to one of Claims 1 to 6 , which is designed to perform the separation in the first separation dimension and the separation in the second separation dimension alternately, in particular only alternately. Sample separating device (10) according to one of Claims 1 to 7 wherein the sample separation device (10) is configured such that, after separating a first portion of the fluidic sample in the first separation dimension, it is intermediately stored in a first sample receiving volume (71), after subsequently separating a second portion of the fluidic sample in the first separation dimension thereof a second sample receiving volume (73) is temporarily stored, and thereafter the first portion and the second portion are sequentially subjected to the separation in the second separation dimension from the respective sample receiving volume (71, 73). Sample separating device (10) according to one of Claims 1 to 8th wherein said means (32; 30, 70) comprises: second sample separation means (32) for separating said fluidic sample separated in said first separation dimension in said second separation dimension; and / or a control device (70) which is designed to separate the fluidic sample separated in the first separation dimension in the second separation dimension by the first sample separation device (30) with a different mobile phase than in the separation of the fluidic sample in the first separation dimension. Sample separating device (10) according to one of Claims 1 to 9 comprising a first fluid valve (84) having fluid ports (86), one to the fluid drive (20), another to the first sample separator (30), yet another to the detector (50), and more particularly to the second sample separator (32) is connected. Sample separating device (10) according to Claim 10 comprising a sample injector (40) for injecting the fluidic sample connected to yet another of the fluid ports (86) of the first fluid valve (84). Sample separating device (10) according to Claim 10 or 11 comprising a second fluid valve (88) connected to a plurality of the fluid ports (86) of the first fluid valve (84). Sample separating device (10) according to one of Claims 1 to 12 , comprising at least one intermediate storage volume (96), which is designed for the temporary buffering of a fluid portion, in particular of a fluidic sample. Sample separating device (10) according to Claim 12 and 13 wherein the at least one intermediate storage volume (96) is connected between fluid connections of at least one intermediate storage valve (94), in particular two intermediate storage valves (94), wherein the at least one intermediate storage valve (94) is fluidically coupled to the second fluidic valve (88). Sample separating device (10) according to one of Claims 1 to 14 comprising a sample fractionator (75, 77) for fractionating separate fractions of the fluidic sample. Sample separating device (10) according to one of Claims 1 to 15 further comprising at least one of the following features: at least one of the first sample separation device (30) and the second sample separation device (32) is designed as a chromatographic separation device, in particular as a chromatography separation column; the sample separation apparatus (10) is configured to analyze at least one physical, chemical and / or biological parameter of at least one fraction of the fluidic sample; the sample separation device (10) comprises at least one of the group consisting of a device for chemical, biological and / or pharmaceutical analysis and an HPLC device; the sample separation device (10) is configured as a microfluidic device; the sample separation device (10) is configured as a nanofluidic device. Method for separating a fluidic sample by means of a sample separating device (10), in particular according to one of the Claims 1 to 16 wherein the sample separation device (10) is operated to separate the fluidic sample in a first separation dimension and to separate the separated fluidic sample in a second separation dimension, the method comprising: operating a common fluid drive (20) to drive the mobile phase fluidic sample in the first separation dimension and in the second separation dimension; Separating the fluidic sample by means of a first sample separation device (30) in the first separation dimension; Separating the fluidic sample separated in the first separation dimension in the second separation dimension; Detecting the separated fluidic sample by means of a detector (50). Method according to Claim 17 wherein the separation of the fluidic sample separated in the first separation dimension occurs in the second separation dimension: by means of a second sample separation device (32); and / or by means of a different mobile phase than in the separation of the fluidic sample in the first separation dimension. Method according to Claim 17 or 18 wherein the method after separation in the second separation dimension comprises returning the fluidic sample separated in the second separation dimension, in particular recirculating an outflow of the detector (50) to the first sample separation device (30) for separating the already twice separated sample in a third one Has separation dimension. Method according to Claim 19 wherein the method after separation in the third separation dimension comprises feeding the already three times separated sample to the second sample separation device (32) for separating the already three times separated sample in a fourth separation dimension.

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