Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/26—Conditioning of the fluid carrier; Flow patterns
  • G01N30/38—Flow patterns
  • G01N30/46—Flow patterns using more than one column
  • G01N30/461—Flow patterns using more than one column with serial coupling of separation columns
  • G01N30/463—Flow patterns using more than one column with serial coupling of separation columns for multidimensional chromatography
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/26—Conditioning of the fluid carrier; Flow patterns
  • G01N30/28—Control of physical parameters of the fluid carrier
  • G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/26—Conditioning of the fluid carrier; Flow patterns
  • G01N30/38—Flow patterns
  • G01N30/46—Flow patterns using more than one column
  • G01N30/461—Flow patterns using more than one column with serial coupling of separation columns
  • G01N30/465—Flow patterns using more than one column with serial coupling of separation columns with specially adapted interfaces between the columns
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/26—Conditioning of the fluid carrier; Flow patterns
  • G01N30/28—Control of physical parameters of the fluid carrier
  • G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
  • G01N2030/322—Control of physical parameters of the fluid carrier of pressure or speed pulse dampers

Definitions

• the flow coupler is configured as a fluidic T-piece, a fluidic Y-piece, or a fluidic X-piece, In case of a fluidic T piece and a fluidic Y piece, two flow streams are combined at one bifurcation point into a single outlet path. In the case of a fluidic X piece, there may be one further fluid conduit. This further fluid conduit can be a second fluid outlet conduit or a third fluid inlet conduit. Other kinds of flow couplers are possible as well.
• the sample separation apparatus comprises a detector for detecting the separated fluidic sample and being arranged in the fluid outlet terminal downstream of the second separation unit.
• a detector for detecting the individual fractions and sub-fractions may be arranged downstream of the second separating unit.
• Such a detector may operate on the basis of an electromagnetic radiation detection principle.
• an electromagnetic radiation source may be provided which irradiates the sample passing through a flow cell with primary electromagnetic radiation (such as optical light or ultraviolet light). In response to this irradiation with primary electromagnetic radiation, there will be an interaction of this electromagnetic radiation with the fluidic sample so that resulting secondary electromagnetic radiation may be detected being indicative of the concentration and kind of fluidic fractions.
• the sample separation apparatus comprises a sample injector for injecting the fluidic sample into a mobile phase and being arranged between the first fluid drive and the first separation unit.
• a sample injector for injecting the fluidic sample into a mobile phase and being arranged between the first fluid drive and the first separation unit.
• an injection needle may suck a metered amount of fluidic sample into a connected loop. After driving and inserting such an injection needle in a corresponding seat and upon switching a fluid injection valve, the fluidic sample may be injected into the path between first fluid drive and first separating unit.
• a mobile phase transported by the fluid drive and constituted by a solvent composition may be mixed with the fluidic sample.
• the first fluid drive is operable with a first flow rate (pumped fluid volume per time interval) being smaller than a second flow rate (pumped fluid volume per time interval) according to which the second fluid drive is operable. Due to the two-dimensional separation procedure, the amount of solvent per time interval pumped by the first fluid drive may be significantly smaller than another solvent composition pumped by the second fluid drive. Also a pressure (for instance a pressure value in a range between 50 bar and 400 bar, e.g. 200 bar) applied by the first fluid drive may be smaller than a pressure (for instance a pressure value in a range between 500 bar and 1500 bar, e.g. 800 bar) applied by the second fluid drive.
• a pressure for instance a pressure value in a range between 50 bar and 400 bar, e.g. 200 bar
• a pressure for instance a pressure value in a range between 500 bar and 1500 bar, e.g. 800 bar
• the first separation unit and the second separation unit are configured so as to execute the respective sample separation in accordance with different separation criteria, particularly in accordance with at least partially but not completely orthogonal separation criteria.
• the term "orthogonal" may particularly denote the conventional but not very accurate understanding that two different separation criteria in a two- dimensional liquid chromatography system relate to completely decoupled parameters. This is not the case in practice, since for instance a separation with regard to mass and a separation with regard to volume of particles such as molecules are not completely decoupled. Exemplary embodiments of the invention make benefit of this cognition and propose to adjust the parameters under a consideration of the fact that the separation criteria of the two separation units are not completely independently from one another.
• the sample separation apparatus may be configured as a fluid separation system for separating components of the sample.
• a mobile phase including a fluidic sample passes through the fluidic device, for instance by applying a high pressure, the interaction between a filling of the column and the fluidic sample may allow for separating different components of the sample, as performed in a liquid chromatography device.
• a gist of an embodiment of the invention is that, instead of coupling both dimensions in an end-to-head fashion, this approach is like stacking one dimension on top of the other. It may sound simple, but the concept is to feed the outlet of the first dimension directly into the high pressure side of the second dimension. In simple terms, if the second dimension needs 800 bar and the first dimension needs 200 bar, then the first dimension pump feeds against 1000 bar. There is a strong advantage due to the fact that now the separated peaks from the first dimension always elute under exactly the same pressure, which is matched to the actual inlet pressure of the second dimension. This now helps that there is now need to pressurize the injected volume, which is switched into the second dimension. So the modulation is a very smooth transition, which is cycling the valve to generate the impulse, which is needed to trigger the chromatographic separation.
• the reason why each of the fractions can further be split into a plurality of sub-sections by the second dimension chromatographic column 208 is that the second dimension chromatographic column 208 may be configured so as to have another separation criterion as compared to the first dimension chromatographic column 204. This may for instance achieved by different chemicals, different solvent composition, different temperature, different size or shape of the two separation systems.
• the two-dimensional liquid chromatography apparatus 200 comprises a first binary pump 202.
• the first binary pump 202 is configured for conducting the fluidic sample to be separated through the first dimension chromatographic column 204.
• the first binary pump 202 provides a mixture from a first solvent 250 (such as water) and a second solvent 252 (such as acetonitrile, ACN).
• a second binary pump 206 which has a significantly higher flow rate as compared to the first binary pump 202.
• the flow rate of the second binary pump 206 may be 4 ml/min, whereas a flow rate of the first binary pump 202 may be 100 ⁇ /min.
• the second binary pump 206 can provide a mixture of a first solvent 254 with a second solvent 256.
• the solvents 254, 256 may or may not be the same as the solvents 250, 252.
• the second binary pump 206 is configured for conducting the already separated or treated fluidic sample conducted via a fluidic valve 218 towards the second dimension chromatographic column 208 which is arranged downstream of the first dimension chromatographic column 204.
• a control unit 238 (such as a processor, for instance a microprocessor or a central processing unit, CPU) which is capable of controlling all the devices and fluidic components shown in Fig. 2. This is illustrated schematically by the dotted lines directed from the control unit 238 towards the corresponding components.
• the control device 238 is also capable of switching the fluidic valve 218.
• the fluidic valve 218 can be switched by the control device 238 so that the first binary pump 202 and the second binary pump 206 remain always in fluid communication with one another, which holds for all switching states of the fluidic valve 218.
• Fig. 2 illustrates a first switching state 260 and illustrates a second switching state 270.
• certain grooves 232 and corresponding ports 230 of the two valve members (a rotor and a stator) are aligned such that the above condition is always fulfilled:
• the binary pump 202 and the binary pump 206 remain always in fluid communication with one another, i.e. are hydraulically coupled.
• Fig. 4 shows a diagram 400 having an abscissa 402 along which a time is plotted and having an ordinate 404 along which a solvent composition as mixed by the first binary pump 202 is plotted.
• the control device 238 is now configured for controlling the first dimension separation column 204 to execute the primary separation sequence 406 as shown in Fig. 4 within a measurement time interval which is denoted with reference numeral 408 in Fig. 4.
• the measurement time interval is 30 minutes.
• the gradient run in accordance with the primary separation sequence 406 is carried out.

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• Physics & Mathematics (AREA)
• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
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• Analytical Chemistry (AREA)
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• General Health & Medical Sciences (AREA)
• General Physics & Mathematics (AREA)
• Immunology (AREA)
• Pathology (AREA)
• Treatment Of Liquids With Adsorbents In General (AREA)

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• 2011
  • 2011-06-20 WO PCT/EP2011/060263 patent/WO2012175111A1/en unknown
  • 2011-06-20 EP EP11743453.0A patent/EP2721403A1/de not_active Withdrawn

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