JP2012520467A - Centrifugal liquid chromatography method and apparatus - Google Patents

Abstract

An apparatus and method for centrifugal liquid chromatography are described. Multiple chromatographic enclosures can be given angular velocity simultaneously. Centrifugal force allows the mobile phase fluid containing the sample to move through the stationary phase within the chromatographic enclosure and perform chromatographic separation on the sample components. By using centrifugation as the driving force, much smaller stationary phase particles can be used compared to high performance liquid chromatography (HPLC). Moreover, when performing equivalent chromatographic separation, separation efficiency becomes very high compared with HPLC by using a centrifugal separation technique.
[Selection] Figure 5A

Description

[Description of related applications]
This application is incorporated by reference in its entirety for all purposes, with the title of the invention “Centrifugal Column Chromatograph System”, March 13, 2009 by Kerr et al. Claims priority under 35 U.SC §119 (e) to filed US Provisional Patent Application No. 61 / 210,118.

  Aspects of the invention relate to a system for performing liquid chromatography. Specifically, this aspect relates to a method and apparatus for performing liquid chromatography in a column subject to centrifugal force.

  Chromatography is a tool used not only by chemists but also by people who apply chemistry to their area of specialization. Chromatography is widely used and essential in the petroleum industry, food industry, pharmaceutical industry, medicine (eg diagnostics) and various other fields. Chromatography is one of the separation processing techniques. The components in the mixture can be physically separated from each other by chromatographic processing. Each physically separated component can be analyzed, for example, for identification purposes. It is also possible to collect the separated components and use them for other purposes. For example, it can also be used as a component that forms other chemical compositions.

  Two important factors in chromatography are separation efficiency and throughput. In chromatographic processing, separation efficiency is related to the ability to separate each component of the mixture, distinguishing each component from each other, and determining whether or not they are chemically the same, if necessary. Ingredients can also be collected. If the separation efficiency in chromatographic processing is inappropriate, it becomes impossible to identify or collect specific components of the mixture that is the subject of the chromatographic processing. On the other hand, the throughput is related to the time required for chromatographic processing, the timing of collecting each separated component, and the time required to collect a specific amount of components. Typically, increasing throughput time increases the cost of chromatographic processing and associated chromatographic systems.

  A common method for performing chromatographic separation is high performance liquid chromatography (HPLC). At present, in the field of liquid chromatography, a chromatographic system using HPLC is excellent in separation efficiency and throughput time. In HPLC, a liquid containing a mixture of components to be separated is moved in a particle bed using pressure as a driving force. As the liquid moves in the particle bed, the components in the liquid are separated from each other by the interaction between the particle bed and the liquid.

  In order to increase the separation efficiency, it is desirable to reduce the particle size of the particles in the particle bed. However, if the liquid does not move sufficiently within the particle bed, the throughput time increases as the particle size decreases. In HPLC, since pressure is used as a driving force, there is a limit to further improvement in separation efficiency. In order to obtain a sufficient throughput time using particles having a particle diameter of about 2 micrometers in diameter, a pressure of about 10,000 PSI is required in HPLC. At higher pressure levels, the particles are more likely to break, and the construction and maintenance of a system that provides higher pressure levels can be very expensive, so increasing the pressure beyond this level has only a limited effect. I can't. Therefore, there is a need for a method and apparatus for performing liquid chromatography that is free from the limitations of HPLC and that can improve separation efficiency and throughput.

  This specification details various embodiments related to systems, methods and apparatus for providing centrifugal liquid chromatography. A rotor is provided that holds one or more chromatographic enclosures. Each chromatographic enclosure contains a chromatographic stationary phase and is configured to provide a flow path through the chromatographic stationary phase. Centrifugal force allows the mobile phase fluid containing the sample to pass through a chromatographic stationary phase within the chromatographic enclosure to perform chromatographic separation processes on each component of the sample. The sample introduction may be controlled so that the flow on the rotor reaches a steady state condition prior to sample introduction. By using centrifugal force as the driving force, it becomes possible to use a very small stationary phase fluid as compared with high performance liquid chromatography (HPLC). Moreover, even when performing equivalent chromatographic separation processing, separation efficiency can be significantly improved by using centrifugal force as compared with HPLC.

  One aspect of the present invention is a centrifugal chromatograph system comprising: 1) a chromatographic enclosure; 2) a rotor that holds the chromatographic enclosure and rotates the chromatographic enclosure; and 3) A sample introduction mechanism fluidly coupled to the chromatographic enclosure. A sample introduction mechanism is configured to introduce sample fluid into the chromatographic enclosure during rotation of the rotor. Further, the sample introduction mechanism receives a sample introduction signal and starts introducing the sample fluid in response to the reception of the sample introduction signal. In a specific embodiment, a control device connected to the sample introduction mechanism may be provided to automatically start the introduction of the sample fluid. In various embodiments, the sample introduction mechanism and / or control device may be retained on the rotor.

  Another aspect of the present invention is a centrifugal column chromatograph system for 1) a rotor configured to rotate about an axis, and 2) for a plurality of chromatographs held in said rotor. A column enclosure; 3) a sample introduction mechanism fluidly coupled to at least a selected one of the chromatographic column enclosures; and 4) fluidly coupled to the plurality of chromatographic column enclosures held by the rotor. And an eluent reservoir containing fluid eluted from the plurality of chromatographic column enclosures.

  In various embodiments, each chromatographic column enclosure contains a corresponding chromatographic stationary phase and allows fluid movement through the chromatographic stationary phase contained within the chromatographic column enclosure. Configured to facilitate. The fluid is moved in the chromatographic stationary phase in the axial direction of the chromatographic column enclosure by centrifugal force generated by rotation of the rotor. A sample introduction mechanism introduces sample fluid into the selected chromatographic column enclosure as the rotor rotates.

  Yet another aspect of the present invention is a centrifugal chromatograph system comprising 1) a rotor configured to rotate about an axis, and 2) at least one chromatograph held by said rotor. An enclosure, and 3) a mixing chamber that is held in the rotor and mixes the mobile phase fluid with the sample on the rotor to produce a mixed fluid. The mixing chamber facilitates the mixing of the sample and the mobile phase fluid by utilizing the rotation of components held by the rotor. Also, the mixing chamber is usually arranged on the upstream side of the chromatographic enclosure. In various embodiments, the centrifugal chromatographic system further comprises a mobile phase fluid reservoir fluidly coupled to the mixing chamber and a sample introduction mechanism fluidly coupled to the mixing chamber, the mobile phase fluid reservoir, The sample introduction mechanism may be disposed either on the rotor or outside the rotor.

  Yet another aspect of the invention is a chromatographic system comprising 1) a chromatographic enclosure containing a corresponding chromatographic stationary phase, the chromatographic immobilization contained within the chromatographic enclosure. A chromatographic enclosure configured to facilitate movement of fluid through the phase, and 2) a rotor holding the chromatographic enclosure, wherein the chromatographic enclosure is rotated at a predetermined angular velocity; A rotor configured to move fluid in the chromatographic enclosure including the chromatographic stationary phase by centrifugal force; and 3) a first portion that is stationary when the rotor rotates. And held on the rotor and shared with the rotor. And a second portion which rotates, and a fluid enclosure in fluid coupled to the chromatographic enclosure.

  In one embodiment, the chromatographic system may include a fluid supply mechanism that is fluidly coupled to the chromatographic enclosure and facilitates the supply of mobile phase fluid to the chromatographic enclosure. The fluid supply mechanism includes a first portion that is stationary when the rotor rotates, and a second portion that is held on the rotor and rotates together with the rotor. The fluid supply mechanism is configured to move fluid from the first portion of the fluid supply mechanism to the second portion when the rotor rotates.

  Another aspect of the present invention is a centrifugal chromatographic system comprising 1) a rotor comprising a chromatographic enclosure held by the rotor, and 2) a gas bearing proximate to the rotor. And a gas bearing configured to stabilize the rotor during rotation of the rotor. The system may further include a containment structure surrounding the rotor and a plurality of rotor support structures supported by the containment structure. Each rotor support structure may include a gas bearing, and the gas bearing may cooperate to stabilize the rotor while the rotor is rotating.

  Another aspect of the present invention is a chromatographic system comprising 1) a plurality of chromatographic enclosures held on a rotor, each chromatographic enclosure having a corresponding chromatographic stationary phase. And is configured to facilitate fluid movement through the chromatographic stationary phase contained within the chromatographic enclosure, and the centrifugal force generated by rotation of the rotor causes A plurality of chromatographic enclosures that move the fluid toward the outer peripheral surface of the rotor, and 2) a plurality of links disposed around the outer peripheral surface of the rotor, each link having two Connected with other links to form a continuous chain ring around the outer peripheral surface of the rotor, A plurality of channels comprising: a link configured to i) contain fluid moving toward the outer peripheral surface, and ii) redirect the fluid inwardly away from the outer peripheral surface A link.

  Another aspect of the present invention is a centrifugal chromatograph system comprising 1) a rotor configured to rotate about an axis and 2) a chromatographic enclosure held on said rotor. A corresponding chromatographic stationary phase and configured to facilitate fluid movement through the chromatographic stationary phase contained within the chromatographic enclosure and caused by rotation of the rotor A chromatographic enclosure for moving the fluid in the chromatographic stationary phase by centrifugal force; and 3) a flow cell fluidly connected to the chromatographic enclosure and comprising a flow window, the chromatographic enclosure comprising: A flow path through the flow cell, the chromatograph Beginning from the predetermined position of the chromatographic within stationary phase down closure, inner cross-sectional area of the flow path through the flow window is substantially constant, comprising a flow cell, a.

  Another aspect of the present invention is a method of operating a centrifugal chromatograph system comprising 1) rotating a rotor holding a chromatographic column, and 2) mobile phase fluid passing through the chromatographic column. A flow is formed, the mobile phase is passed through the chromatographic column by centrifugal force, and 3) the sample fluid is introduced into the chromatographic column by introducing the sample fluid into the flow of the mobile phase fluid. The sample fluid is introduced while the rotor rotates. The method further determines whether or not the interior of the chromatographic column has reached a steady flow condition of the mobile phase fluid, and determined that the interior of the chromatographic column has reached the steady flow condition of the mobile phase fluid. Later, the sample fluid may be released. In addition, the method further introduces a sample fluid into the chromatographic column after it is determined that a steady flow condition of the mobile phase fluid has been reached, thereby facilitating centrifugal chromatographic separation of the sample fluid. It may be.

  Yet another aspect of the present invention is a method of operating a centrifugal chromatograph system comprising 1) rotating a rotor holding a chromatographic column containing a chromatographic stationary phase, and 2) rotating the rotor. In the meantime, the mobile phase fluid is supplied to the chromatographic column, and the mobile phase fluid is passed through the chromatographic stationary phase by centrifugal force. 3) Timing of introducing the sample fluid into the chromatographic column To facilitate centrifugal chromatographic separation of the sample fluid.

  Other aspects and advantages of the present invention will become apparent from the following detailed description, taken in conjunction with the accompanying drawings, illustrating by way of example the principles of the invention.

  The invention may be further understood by the following detailed description with reference to the accompanying drawings. In the following description, the same reference numerals indicate the same structural elements.

1 is a block diagram illustrating a chromatographic system including a rotating element.

FIG. 3 is a side view illustrating an example of a rotor assembly configured for a chromatographic system.

FIG. 3 is a top view showing an example of a rotor assembly configured for a chromatographic system.

FIG. 3 is a top perspective view of a manifold assembly configured for a chromatographic system.

Bottom view of a manifold assembly configured for a chromatographic system.

FIG. 3 is a vertical cross-sectional view of a rotor assembly including a flow path illustration.

FIG. 3 is a top perspective cross-sectional view of a dish-like assembly, a chromatographic column, a flow cell, a return segment link, and a return channel including flow path illustrations.

The perspective view of a return path segment link.

FIG. 3 is a cross-sectional view of a reservoir and a side view of a rotor assembly.

FIG. 3 is a side view of a gas bearing support assembly.

FIG. 3 is a side view of a gas bearing support assembly.

FIG. 3 is a perspective view of a gas bearing support assembly.

The side view which shows the rotor assembly and gas bearing assembly which are arrange | positioned inside a containment structure and are provided with an instrument attachment part.

FIG. 3 is a top view showing an example of a rotor assembly configured for a chromatographic system.

FIG. 3 is a side view illustrating an example of a rotor assembly configured for a chromatographic system.

The block diagram which shows the structure with which the reaction chamber was provided between the column edge part and the flow cell.

FIG. 2 is a front view and side view of a bucket assembly comprising a plurality of columns for performing chromatographic separation.

FIG. 3 is a top view showing a rotor assembly including a plurality of buckets in rotation.

FIG. 3 is a top view of a rotor assembly that includes a plurality of buckets at rest.

FIG. 3 is a top view showing a rotor assembly including a plurality of buckets in rotation.

The front view of the column set for the chromatographic processing before centrifugation and after centrifugation.

The figure which shows the chromatogram which isolate | separated three types of components.

The flowchart which shows the method for performing the chromatographic separation method.

  Embodiments of the present invention will be described in detail with reference to the accompanying drawings. The following description does not limit the embodiments of the present invention to preferred examples. Various changes and modifications can be made within the scope of the present invention described in the claims, and equivalents thereof are also included in the scope of the present invention.

  Chromatography means the process of physically separating a chemical mixture into its components. In chromatographic processing, a mixture of chemicals is dissolved (or mixed) into a carrier stream (gas or liquid). The carrier stream containing the mixture is passed through a bed of particles. The carrier stream moves through the bed of particles at a certain flow rate. In chromatography, the carrier stream is often called the “mobile phase” and the bed of particles is called the “stationary phase”.

  The stationary phase particles are selected such that the components of the mixture dissolved in the mobile phase interact to a different extent than the stationary phase particles. The difference in the extent to which the components of the mixture interact with the particles affects how quickly each component of the mixture passes through the stationary phase. Thus, the components of the mixture pass through the stationary phase at different rates. When the two components of the mixture pass through the mobile phase at different speeds, the component that passes through the stationary phase faster than the component that passes through the mobile phase more slowly during a certain period of time, Because it moves a lot, it is separated.

  The purpose of liquid chromatography is to provide a system that includes a selected mobile phase and a selected stationary phase such that the components of the mixture pass through the mobile phase at different rates. When the components of the mixture are separated at least to some extent, various types of measuring devices are used to characterize the nature of the separated components and in some cases to quantify the components of the mixture. In addition, if necessary, components separated from the mixture are collected for further processing or analysis.

  In liquid chromatography, as described above, the mobile phase passes through a stationary phase containing particles such as spherical particles having a certain radius. The particles can have pores and other molecules bind to the particles. The particle radius, pore size, and binding molecules can be selected to have a different type of physical interaction with the components dissolved in the mobile phase.

  In the earliest examples of liquid chromatography, gravity was used as the driving force for the mobile phase to pass through the stationary phase. For example, the mobile phase was loaded on top of an open column packed with particles of relatively uniform particle size distribution forming a stationary phase. As the mobile phase passes through the mobile phase under gravity, the components of the mixture added to the mobile phase are separated. In some cases, since the separated components have different colors, it can be visually observed that the components are separated. When the column is made of a transparent material such as glass, it can be observed that different colored bands descend the column. The column used to pack the particles and thus separate the components in the mixture is referred to as a “chromatographic column”.

  Since liquid chromatography is a developed field, the ability to separate components in a mixture using a chromatographic column has been found to increase as the size of the stationary phase packed particles decreases. The use of smaller particle size improves the packing efficiency of the stationary phase and increases the surface area in the volume where interaction occurs between the components of the mixture and the stationary phase particles. Therefore, a method has been developed for producing smaller spherical particles for use in the stationary phase of chromatographic columns.

  In a chromatographic column driven by gravity, the particle size in the mobile phase cannot be reduced indefinitely. Gravity has a particle size limit that no longer allows a mixture of mobile phase and components to pass through the stationary phase in a practical time. To overcome the limitations of using gravity as the driving force, other driving forces, such as pressurization, are used to pass the mobile phase through the chromatographic column.

  High performance liquid chromatography (HPLC) has been developed so that particles of smaller size can be used for the stationary phase. This increased the separation performance of the chromatographic column. In HPLC, high pressure is used as a driving force for passing the mobile phase through the chromatographic column. In HPLC, the pressure required to pass a mobile phase through a chromatographic column is inversely proportional to the square root of the particle diameter. In order to use a particle size of a few microns, the HPLC system needs to be configured to operate at a pressure of approximately 10,000 PSI, thus requiring a high performance and expensive instrument. Currently, pressure resistance, the use of smaller particles to improve separation efficiency, is a limiting factor for further development of HPLC.

  In order to improve the drawbacks related to the pressure resistance of the HPLC system, it has been studied to use another driving force to pass the mobile phase through the stationary phase in the chromatographic column. As already mentioned, gravity was used as the driving force in early chromatographic systems. The effect of gravity on the object can be simulated and the effect of gravity can be increased by placing the object on a platform such as a disk that rotates about the axis of rotation. As an object moves in a rotating coordinate system, the object receives a force along a line perpendicular to the axis of rotation that acts like gravity.

The force received by the object in the rotating coordinate system can be called “centrifugal force”. Centrifugal force is often expressed as a multiple of gravity. For example, Relative Centrifugal Force (RCF) is a measure of acceleration applied to an object that rotates about an axis of rotation and is expressed in units of “gravity” or g.
RCF = r (2πN) ² / g
Here, r is the radius of rotation, that is, the distance from the rotation axis to the object, N is the rotation speed expressed in number of rotations per unit time, and g is the gravitational acceleration. The RCF can also be written as
RCF = 1.118x10 ^-5 r _cm N ² _RPM
Where r is the radius of rotation measured in cm and N ² _RPM is the rotational speed expressed in revolutions per minute.

  As will be shown below, RCF can be used as a driving force for passing a mobile phase through a stationary phase packed in a column for a carmatograph. In order to use RCF, the mobile phase passes through the stationary phase of the column for the chromatograph, whereas the column for the column is rotating. For example, a column for a carmatograph is installed on a rotating disk, and a chromatographic operation is performed while rotating the column. An apparatus and method for using RCF in a chromatographic system is shown in the following figure.

  Initially, a chromatographic system having one or more rotating elements is shown in FIG. Next, details of the various forms of the chromatographic system are shown in FIGS. In particular, specific examples of rotating parts for a chromatographic system are shown in FIGS. Specific examples of rotating parts support and supply the flow paths of many chromatographic enclosures during rotation. The “hanging bucket” type is shown in FIGS. 10A to 10D. In the hanging bucket type, the chromatographic enclosure can be detached from the rotating body more easily than the embodiment shown in FIGS. 2 to 9C. An example of a chromatographic enclosure before and after centrifugation is shown in FIG. The chromatogram is shown in FIG. Finally, FIG. 13 shows a method for performing the chromatographic separation process.

Chromatographic System FIG. 1 is a block diagram of a chromatographic system 100 that includes a rotating element 132. In many embodiments, the rotating element is also referred to as a “rotor”. The rotating element 132 can be used to apply RCF to the components of the chromatographic system. Before discussing the use of RCF in the chromatographic system 100 and the effects associated therewith, some components of the chromatographic system 100 will be described. The description of the chromatographic system and the number of components included in the chromatographic system are not intended to limit the invention to these examples in any way, but only for illustrative purposes. An embodiment of a chromatographic system having a rotating element may include other components than those shown in FIG. 1, or may include each component in a different arrangement.

  The chromatographic system 100 may include a solvent management unit 108. The solvent management unit 108 allows different solvents to be used for various chromatographic operations and can be changed for each operation of the chromatographic system. The solvent management unit 108 may include a solvent reservoir such as 102. The solvent reservoir can be used as a source for supplying a mobile phase in which the sample is dissolved for chromatographic operations. Depending on what type of sample is analyzed, a single solvent or a combination thereof is used. Furthermore, the solvent used can be changed for each measurement. That is, the first solvent may be used for chromatographic analysis of the first sample in the first measurement, and the second solvent may be used for analyzing the second sample in the second measurement. . A flow control mechanism such as a valve allows different solvents to be used at different times depending on the individual measurement requirements.

  A solvent supply system such as 104 can be used to move the solvent from the solvent reservoir to form a mobile phase. Typically, one pump or some type of pump can be used to provide the driving force to move the solvent from the solvent reservoir 102. The gradient forming unit 106 can be used to generate a mobile phase composed of one or a plurality of solvents. Two-component solvent mixtures are usually used, but more complex solvent mixtures can be used in the examples shown. If the mobile phase is formed from a combination of organic solvents during a single run of the chromatograph, the concentration of each solvent can be varied as a function of time. The gradient forming unit 106 can control the concentration of the components of the mobile phase as a function of time.

  The chromatographic system 100 may include a sample management unit 114. The sample manager 114 introduces the sample into the mobile phase before the mobile phase is introduced into the chromatographic system. The sample management unit 114 has a mechanism for holding various samples 110 and filling the sample injector 112 with the samples. A sample injector is one example of a mechanism for introducing a sample. Sample injectors such as 112 can be used to introduce specific samples into the mobile phase. For example, a sample injector can inject a sample into a mobile phase stream moving in a conduit.

  The chromatographic system 100 may include a column management unit 117. The column manager 117 can be used to control conditions relating to the chromatographic column such as temperature. The temperature of the chromatographic column can be controlled using a device that heats or cools the column, for example by a thermoelectric element (Peltier element) or some other method of adding or removing thermal energy from the column. . A temperature gradient along the column will change the viscosity along the column, resulting in a change in velocity along the column. Typically, as a result of heat conduction through the wall, the temperature is highest near the center of the column and decreases towards the column wall. By heating the wall of the column, the temperature change from the center of the column toward the wall can be reduced, and the temperature of the entire column can be made uniform. Temperature is known to dramatically affect chemical equilibrium, such as the equilibrium between analyte, mobile phase and stationary phase. In order to obtain highly reproducible chromatographic results, it is important to keep the inside of the column at a constant temperature.

  The column management unit 117 may include software that holds the characteristic history of each column. Its properties include 1) when it was packed, 2) what is the composition of the packing, ie what is the stationary phase, 3) how many times the column has been used, and 4) the column like the composition of the mobile phase solvent. However, the present invention is not limited to these types of chromatographic operations.

  The chromatographic system 100 may include a detection management unit 120. The detection management unit 120 can control various devices for characterizing the components separated from the mobile phase by the chromatographic processing inside the chromatographic column. For example, one or more to detect changes in the amount of light emitted from a light source (UV to visible region, 190 to 700 nm) as light passes through a flow cell window through which eluate from the column passes. A spectrophotometric detector is used. The components physically separated from the mixture are still dissolved in the mobile phase but pass through the flow cell. There, the light emitted from the first light source passes through the first window of the flow cell and interacts with the components. Light can exit the flow cell through the second window of the flow cell where it is collected. The collected light can be used to determine whether there has been an interaction between the light and the separated component.

  The chromatograph system 100 may include a data management unit 115. A data manager 115 can be provided to collect, analyze and store data obtained from one or more detectors. A data manager 115 can also be provided to output data obtained from the detector. For example, the data management unit 115 can be set to output the chromatogram to the display of the carmatograph system. The data management unit 115 may be configured to be able to track and store information collected from different chromatographic columns for data relating to various parameters of individual chromatographic measurements.

  The chromatographic system 100 may include a collection management unit 124. The collection management unit 124 may include a fraction collector 122 for collecting components separated by passing through the column 116. Some components of interest elute the column at different times and the collection manager 124 can be set to direct each of the two or more components to the fraction collector. Some components that elute from the mobile phase (referred to as eluates) are not considered and may be considered “waste”. For example, the solvent can be passed through the column before introducing the sample. The collection management unit 124 can collect the solvent eluted from the column before introducing the sample into a collection device such as the waste liquid collector 123.

  In the embodiment described herein, the chromatographic system may comprise one or more rotating elements, such as rotating element 132. The rotating element is controlled by the rotor management unit 140. The rotor management unit 140 controls the rotation speed of the rotation element 132 with respect to time, for example, an increase in the rotation speed, a steady rotation, a decrease in the rotation speed, and the like. The rotor management unit 140 can monitor whether or not the rotating element 132 is operating correctly, and can perform a process associated with the operation of the rotating element 132 such as automatic adjustment. The rotor manager 140 can also monitor and control the power supply to many elements operating on the rotating element 132, such as an electronic control valve on the rotating element.

  The system manager 103 can be configured to monitor and control the overall functionality of the system during various operating modes of the chromatographic system 100, such as initialization mode, operating mode, and stop mode. it can. The system management unit 103 communicates with the flow management unit 101, the solvent management unit 108, the sample management unit 114, the data management unit 115, the column management unit, the detection management unit 120, the collection management unit 124, and the rotor management unit 140. Can be set to send commands. The system management unit 103 can also be set to exchange information with other devices and systems such as other chromatograph systems and remote computers.

  The chromatographic system 100 can manage one or more flows. The flow management unit 101 can be set to control many valves and pumps in the system 100 directly or by communication using other device components. For example, the flow management unit 101 sends a command to the solvent management unit 108 to supply a specific solvent at a predetermined flow rate, and one or more logic devices related to the solvent management unit 108 are supplied at a predetermined flow rate. Devices such as valves and pumps may be controlled as is done. Alternatively, the flow rate management unit 101 may directly control valves and pumps related to the solvent management unit 108.

The flow path management and flow analysis flow management unit 101 establishes and holds a plurality of flow paths so that the number of flow paths can be changed for each system or even within one system. The flow in the chromatographic system 100 may begin at the beginning 126 of the flow path. Specifically, it may start from the solvent reservoir 102. Fluid can be moved from the reservoir to the gradient former 106 via the solvent supply system 104. At 134 points in the flow path, the flow is moved to the rotating element 132. Flow movement occurs when the rotating element 132 is rotating in the direction of rotation 138, which is a certain rotational speed. In different embodiments, the rotational speed and direction of rotation can be varied. In other embodiments, the flow rate varies but the rotating element is stationary.

  On the rotating element 132, the flow at various locations is either outward from the center of the rotating element 132, or otherwise toward the center of the rotating element 132. Along the flow path on the rotating body, the flow can also be changed according to the level of the rotating element. For example, in the first stage the flow moves through the chromatographic column and in the next stage, for example, it flows to a reservoir under the chromatographic column.

  In certain embodiments, the flow may enter near the center of the rotating element, flow out of the center, and enter chromatographic columns such as 116a, 116b and 116c. For example, the flow starts from a common source near the center, such as a mixing chamber located near the center, and branches into a plurality of channels. For example, the flow may be branched when passing through the sample management unit 114, and different samples may be introduced into different flow paths. This example of a branch location is for illustration only. In certain embodiments, the branching point at which the flow is divided into a plurality of flows is defined inside the solvent management unit 108, that is, before or inside the reservoir 102, the solvent supply system 104, before or inside the gradient forming unit 106, and the sample management unit. It may be placed at any position in the flow path such as before or inside 114, before or inside column 116, before or inside detection manager 120, or before or inside collection manager 124. Furthermore, the flow management unit 101 can be configured to change the branch point for each measurement of the chromatograph system.

  As an example of the branching of the flow path, the flow path may start from one flow path before the sample management unit 114. In the sample management unit 114, the flow path is branched into a plurality of flow paths 130, for example, three flow paths. If necessary, a different sample may be injected into each flow path and pass through one of the chromatographic columns 116a, 116b, and 116c.

  In another example, the flow can branch after the sample manager 114. The single channel enters the sample management unit 114, and the sample is injected into the single channel. After the sample management unit 114, it is also possible to divide the flow path into a plurality of channels and to process the branched flows in parallel by a plurality of chromatographic columns. For example, one flow path containing a sample can be generated by the sample management unit 114, branched into three flow paths, and processed by the chromatographic columns 116a, 116b, and 116c.

  In one embodiment, the flow management unit 101 of the chromatographic system 100 can be configured to change or control the number of flow paths and branch locations. The flow management unit 101 may include a pipe line in which many flow paths are formed at the same time. Furthermore, the flow management unit may be provided with a switch mechanism such as a valve that can be opened and closed at different positions and can change the number of flow paths formed at specific positions.

  For example, the flow management unit 101 can control a pipeline in the sample management unit 114 that can create up to three flow paths. The flow management unit can control branching mechanisms located before and after the sample management unit. In one manner, three channels are created before the flow reaches the sample manager 114 (each channel is joined to a chromatographic column such as 116a, 116b and 116c). In each of the three flow paths, the sample management unit 114 can inject a different sample, and the sample proceeds to a chromatographic column for analysis.

  In another aspect, the flow manager turns off the branching mechanism before the sample manager 114, so that one flow enters the sample manager 114 and one sample is injected. After the sample is injected into one channel, the branching mechanism is activated and one flow is branched into a plurality of channels and passes through a plurality of columns. For example, one flow path may be branched into three flow paths so as to be processed by three columns 116a, 116b, and 116c.

  The flow management unit 101 controls the flow switch mechanism and different combinations of flow paths at various points in the chromatographic system 100, for example, in the solvent management unit, in the sample management unit 114, in the column management unit 117. The detector management unit 120 or the collection management unit 124 can be integrated or branched. For example, when three flow paths are possible, the flow management unit can control the switch mechanism so that a single flow path, three flow paths, or two flow paths can be generated at different times in the chromatographic system 100. 101 can be set. Further, at another location within the flow path, the flow can be changed from a single flow path to a two flow path, from a single flow path to a three flow path, or from a two flow path to a three flow path.

  After the flow path has passed through each column, it is analyzed using one or more different detectors 118. For example, the flow cell can be placed near the end of the column and its transparent window allows the light source to be seen through the flow cell. While the flow cell passes through the column eluate (the component in which the initial sample mixture is physically separated from the solvent mixture), the light exiting the flow cell is detected using a detector 118 such as a high electron multiplier. Be captured. In another example, after passing through the chromatographic column, a portion of the stream is directed to a device such as a mass analyzer for additional analysis.

  In one embodiment, a single detector may analyze a plurality of flow paths. With one light source and one photomultiplier tube, it is possible to analyze a flow passing through a plurality of flow cells such as three flow cells provided in each of the chromatographic columns 116a, 116b and 116c. Costs can be reduced by sharing the apparatus with multiple flow paths. Further details regarding device sharing are shown in FIGS. 10A-10D.

  At another point in the flow path, such as 136, the flow exits the rotating element 132. While the rotating element 132 is rotating and the rotating element is stopped, the outlet 136 from the rotating element functions. As shown, the flow can enter the waste reservoir or fraction collector after leaving the rotating element 132. The flow entering the waste reservoir and / or fraction collector ends at the end 128 of the flow path.

  In addition to the branching of the flow path, the flow path also merges. FIG. 1 shows an example of flow path merging in which a plurality of flow paths merge into one flow path. For example, if the sample is not collected, all the outflow channels from the chromatographic column merge into one channel and go to a common waste reservoir. As in the case of the branching of the flow path, the chromatograph system 100 has a flow path switching mechanism, and a plurality of different flow paths may be joined at different positions for each measurement. The flow path management unit 101 can be set to perform flow path control related to merging, such as controlling a place where merging occurs.

One aspect of the column condition control unit chromatograph system 100 is that the conditions can be established repeatedly inside the chromatographic column. That is, the system 100 can be set to establish and maintain specific conditions for a chromatographic column for a specific flow path. Conditions to establish and maintain include: 1) the flow rate inside the column, 2) the solvent composition that changes over time in the chromatographic process, 3) the column temperature, 4) the solvent temperature, and 5) the column's angular velocity. Although there are rotation conditions, it is not limited to these.

  In certain embodiments, the chromatographic system 100 can be configured to establish an initial steady state condition within the chromatographic column, such as a constant flow rate, before injecting the sample into the chromatographic column. Establishing an initial steady state refers to bringing each selected combination of column parameters into a state that changes within a certain tolerance over a time range. One reason for reaching a steady state before injecting a chromatographic sample is reproducibility. Chromatographic experiments can be repeated for a variety of reasons, and it is desirable that the experiments be performed under the same conditions each time.

  For example, chromatographic experiments are repeated to collect fractions. Under certain conditions, a sample component in a chromatographic column may depend on the interaction within the chromatographic column, and the length of time that the sample component remains inside the column that remains inside the column for a certain amount of time is Depending on the column conditions (Chromatograph column conditions are intentionally selected so that the difference in time during which one component and another component are held inside the column is large. Reflects the chromatographic separation efficiency of the column). The components that elute from the column can be collected in a certain time. A part of the flow collected from the column at a certain time is called a fraction. The advantage of establishing steady state conditions in the chromatographic column prior to sample introduction is that it is possible to set up a repetitive operation to collect fractions at a fixed time after sample introduction.

In another example, chromatographic processing can be used to determine the presence and amount of specific components in a sample. The chromatographic operation can be repeated many times to account for statistical errors associated with the measurement, such as the amount of components in the sample. As a guide, the sampling error is proportional to 1 / N ^1/2 (eg, 100 samples are required for a 10% error). Therefore, by repeating the chromatographic operation for a specific sample component many times, it is possible to suppress an error associated with the measurement, such as the amount of substance of the component, within a reasonable range. The advantage of establishing a steady state in the chromatographic column prior to sample introduction is that it minimizes errors associated with transient effects that occur with each run.

  In connection with FIG. 1, many different factors are involved in establishing a steady flow condition in a column for a particular flow path. In some embodiments, this function can be controlled by the flow manager 101. To establish a steady state, the flow manager 101 receives data and provides instructions to many elements of the chromatographic system 100.

  In one example, the system 100 initializes operation to establish a steady state flow, and then a rotating element such as 132 increases the rotational speed to a constant angular rotational speed according to the angular rotational speed profile. . The flow management unit 101 starts the flow on the rotating element 132 while increasing the rotation speed or reaching a constant speed. To initiate a flow, the flow manager directs the solvent supply system. The solvent supply system is above or otherwise on the rotating element 132 and begins to introduce solvent into the flow path. The solvent can then begin flowing through the system 100, such as chromatographic columns such as 116a, 116b and 116c.

  In one embodiment, to determine whether the mobile phase has reached steady state velocity, the flow manager 101 instructs the solvent manager 108 of the proportion of the first component of the solvent that changes in a known manner. To do. Thereafter, for example, the proportion of the first component increases or decreases as a function of time. As the solvent component that changes, a solvent component that does not interact with the chromatographic column and that can be detected by one of the devices of the detection management unit 120 is selected. Using the information on how the first component changes and the information received from the detection manager 120, the flow manager measures the mobile phase velocity and its change over time.

  In another example, a mass flow meter can be placed in a flow path, such as after a flow cell. A mass flow meter can be used to measure the flow rate. Based on the information received from the mass flow meter, it is determined whether system components such as the flow management unit 101 have reached the speed of the steady state mobile phase. If the mobile phase velocity and other column conditions are within acceptable limits and the change over time is determined to be within acceptable limits, then the column is determined to be ready for sample introduction.

  As described above, the rotating element 132 may comprise a number of chromatographic columns such as 116a, 116b and 116c. The determination of whether a steady state flow and / or column state has been reached before sample injection can be made for each column. At least flow / column conditions determined prior to sample injection include mobile phase velocity, solvent composition, channel pressure (eg before and after column), flow temperature (eg before and after column), column temperature (eg outside column) And combinations thereof, but are not limited to these. As described above, information about each column, such as whether or not the steady state of the column has been reached, is stored by one or more system components such as the column management unit 117.

  Measurement of flow and column conditions can be measured as a function of time after the sample is introduced. For example, the flow rate can be measured while the sample travels through the chromatographic column. In another example, one or more outside column temperatures can be recorded along the column as the sample travels through the chromatographic column.

  In certain embodiments, the chromatographic system 100 can be configured to determine whether the flow and / or column conditions during the chromatographic operation are within acceptable limits. A certain period of time can be taken during the chromatographic operation, and the column state can be observed during that time. Before or after introducing the sample, unacceptable conditions may occur, such as not reaching steady state. For example, one or more columns may deviate from acceptable ranges at some point in the mobile phase velocity or temperature or in the course of parameter changes. A flow cell defect associated with one of the columns (eg, one of the windows is contaminated), a temperature cell defect associated with one of the columns, a pressure sensor defect associated with one of the columns, or one of the flow paths. One or more parameters may be determined to have exceeded an acceptable range due to a number of factors, such as a leak at (a pressure sensor is used to determine if there is a leak).

  Based on the flow and column conditions determined during each measurement, the system 100 may distinguish between acceptable and unacceptable columns. A predetermined column may be determined as an unacceptable column before sample introduction. For example, when it is determined that a certain column is not in a steady state, the column may be determined to be unacceptable. Another column may be determined as an unacceptable column after sample introduction. For example, when it is detected that the column pressure exceeds the allowable range while the sample passes through a certain column, it may be determined that the column is not allowable.

  In some embodiments, if some of the columns used by the system 100 are determined to be unacceptable, the data collected from the columns determined to be unacceptable is ignored and the data collected from the columns determined to be acceptable is ignored. You may make it use only. For example, the first chromatographic column such as 116a is determined to be unacceptable in any of the following cases. 1) The initial steady state condition is not within the acceptable range. 2) A steady state condition is met, but a parameter is outside the acceptable range. 3) When the sample passes through the chromatographic column, the flow condition or column condition deviates from the allowable range. On the other hand, the second chromatographic column, such as 116b, satisfies the steady-state condition, and all values are acceptable both before and after sample introduction, ie, throughout the chromatographic process associated with a particular measurement. If it is within the range, it is determined as an acceptable column. Similar to determining whether steady-state conditions are met, when determining whether each column is acceptable or not for the purpose of using data collected from the column, the determination should be made for each column. Can do.

System Functions on or Outside the Rotating Element In various embodiments , the functions of the chromatographic system 100 may be performed by components disposed on the rotating element, such as 132, or It may be performed by a component arranged outside the rotating element. Whether a particular function is performed on a rotating element or outside a rotating element can vary depending on the system and can vary from measurement to measurement within the same chromatographic system. The rotating and non-rotating elements of the system shown are for illustration only and are not limited to the example shown in FIG.

  In FIG. 1, a part of the flow path and a part of the system accompanying it are arranged on the rotating element 132, and a part of the flow path and a part of the system accompanying it are arranged outside the rotating element 132. For example, the solvent management unit 108 and the collection management unit 124 are arranged outside the rotation element 132, and the sample management unit 114, the column management unit 117, and the detector management unit 120 are arranged on the rotation element 132. In another embodiment, the entire chromatographic system 100 including the solvent manager 108, sample manager 114, column manager 117, detector manager 120, and collection manager 124 is placed on a rotating element such as 132. It may be done. Furthermore, a part of the chromatographic system 100 may be installed on both the rotating element and the non-rotating element in an overlapping manner. For example, among the solvent management units provided in the chromatographic system 100, a first solvent reservoir may be provided on the rotating element, and a second solvent reservoir may be provided on the non-rotating element. In some cases, a first solvent reservoir disposed on the rotating element 132 may be used, and in other cases, a second solvent reservoir disposed outside the rotating element 132 may be used. In yet another embodiment, both a first solvent reservoir disposed on the rotating element 132 and a second solvent reservoir disposed outside the rotating element may be used.

  In another example, the detection management unit 120 includes a first device such as a mass analyzer on the rotating element 132 and a second device such as a light source and a photomultiplier tube outside the rotating element 132. An apparatus may be provided. Furthermore, even if the light source and photomultiplier tube are located outside the rotating element 132, the flow cell necessary to use this device can be retained on the rotating element 132. In some embodiments, only a first device disposed on the rotating element 132 is used. In another embodiment, only a second device located away from the rotating element is used. In still another embodiment, a combination of devices disposed on and outside the rotating element, such as the first device and the second device, may be used.

As described on the stationary-rotating interface , the chromatographic system may include one or more rotating elements such as 132. At various times during the operation of the chromatographic system, one or more rotating elements such as 132 can be stopped or rotated. It is desirable to provide an interface that allows some amount of movement between the stationary and rotating elements when the rotating element is rotating (during rotation). Examples of such quantities include fluid (eg, gas or liquid), power and data. One rotating element is connected to the stationary element via a plurality of stationary-rotating interfaces. For example, one rotating element is connected to many different stationary elements via multiple fluid interfaces.

  In certain embodiments, the interface can be designed to move the amount in one direction, for example, from a stationary element to a rotating element or from a rotating element to a stationary element. For example, the first interface can be configured to deliver fluid from the stationary element to the rotating element, and the second interface can be configured to deliver fluid from the rotating element to the stationary element. In other embodiments, the interface can be set to be bi-directional and the fluid can be moved from a stationary element to a rotating element and from a rotating element to a stationary element at the same time or at different times.

  For example, one fluid interface can be configured to move fluid from a stationary element to a rotating element while simultaneously receiving fluid from the rotating element and moving it to the stationary element. In this example, the flow can be moved between a stationary element and a rotating element using a separate conduit. In another embodiment, one conduit initially moves fluid from the stationary element to the rotating element, and in the second stage, the conduit receives fluid from the rotating element and moves it to the stationary element.

Mode of Operation Like the system 100 of FIG. 1, the illustrated chromatographic system can be set to operate in many different modes. During different operating modes, the chromatographic system can perform different functions. Examples of different operating modes include initialization, operation, spin up processing, spin down processing, chromatographic processing and data collection between each measurement, and malfunction.

  During initialization, the chromatographic system can self-diagnose the status of many system elements. Examples of conditions to be self-diagnosed include: device status, fluid reservoir height status, gas pressure status, pump status, valve status, motor status, balance check, device energy status, and The state of information communication between devices can be mentioned, but is not limited thereto. These self-diagnostics include other system elements, for example, flow management unit 101, solvent management unit 108, sample management unit 114, data management unit 115, column management unit 117, detection management unit 120, collection management unit 124, and It may be related to the rotor management unit 140. After the self-diagnosis is made, it is determined whether the chromatographic system enters an operating mode or whether one or more chromatographic system components need modification. The chromatographic system can indicate whether correction is necessary, or it can indicate that the chromatographic system is in operation.

  Once the chromatographic system is in operation, settings may be made for individual measurements. In some embodiments, the chromatographic system allows the user to specify one or more adjustment parameters for operation, such as via the system manager 103. Once the parameters for an individual measurement are specified, the chromatographic system performs an operation using the selected parameters. For example, the chromatograss system may spin up one or more rotating elements, such as element 132, from a rest state to a predetermined target angular speed according to a predetermined angular velocity profile. Here, the “spin-up process” includes a rotation management function such as a balance check and adjustment of the system, and a flow management function such as initial fluid introduction to the system and leak detection.

  After the spin-up process, the system determines whether a steady flow state or the like has been reached and the chromatographic operation can be started. When the chromatographic operation is ready, functions such as sample introduction and data collection can be started. The system records the chromatographic progress until it is determined that the operation is complete. When the run is complete, the chromatographic system begins the spin down process.

  During the spin down process, the rotating element 132 decelerates from a specific angular velocity to a stopped state according to a certain angular rotation profile. Prior to or during deceleration, the flow manager 101 can change the flow function such as stopping the flow in the rotating element 132. After the spin down process, the rotation is at rest, while the discharge from the solvent reservoir on the rotating element and the removal of the fraction collected on the rotating element works. During operation, functions such as flushing fluid from the column of rotating elements for cleaning work. Column outflow includes spinning up and spinning down rotating element 132 to expel fluid from the system.

  Any operating mode can cause malfunctions. When a malfunction is detected, an inherent malfunction mode is entered depending on the type of malfunction. For example, if a leak is detected during the spin-up process, or if the balance suddenly changes while the rotating element is rotating, the spin-down malfunction mode is activated. In the spin down malfunction mode, the rotating element can be decelerated faster than it is decelerated by normal operation.

Rotating Elements FIGS. 2-8 show details of one embodiment of a rotating element used in a chromatographic system. Another embodiment of the rotating element is shown in FIGS. 9A-9C. In particular, FIGS. 2 and 3 show a rotating assembly that includes a number of chromatographic columns. With reference to FIGS. 4A, 4B and 6, the various elements of the rotating assembly will be discussed. 5A-5C show further details of the rotating assembly, including a discussion of the flow path through the rotating element. In FIGS. 7A-7C and FIG. 8, many elements are discussed in which the rotating assembly can be utilized as part of a chromatographic system. 9A, 9B, and 9C show an example of a rotor assembly that includes sample injection, column conditioning, power generation, off-rotor communication, fraction collection, reaction chambers, and the like. 10A to 10D show another embodiment of centrifugal liquid chromatography including a hanging bucket type.

  2 and 3 show side and top views of one embodiment of a rotating assembly used in a chromatographic system as shown in FIG. A rotating assembly is one embodiment of a rotating element. As shown in FIG. 1, the elements above or outside the rotating element vary from one chromatographic system to another. Accordingly, the embodiment shown in FIGS. 2-9C is merely illustrative.

  The rotating assembly 200 includes a liquid introduction assembly that includes a pan assembly 202, an adapter plate assembly 214, and a mixing chamber 228 located in the center of the pan assembly 202. Manifold assembly 204 includes a disk portion 206 and a column portion 208. The liquid introduction assembly can be secured to the pan assembly 202 with a number of fasteners. The dish-like assembly 202 is secured to the disk portion 206 of the manifold assembly with fasteners. The bottom of the column portion 208 of the manifold assembly can be secured to the adapter plate assembly 214 via an adapter plate (see FIG. 6 for details of the adapter plate assembly 214 and adapter plate).

  The reservoir 210 is attached to the adapter plate assembly 214. The adapter plate assembly includes an interface to the shaft, such as a tapered shaft 212. The shaft includes a mechanism that attaches to a motor (not shown) that transmits angular rotational speed to the rotating assembly. The angular rotational speed of the rotating assembly can be varied as a function of time based on operational commands sent to the motor. In one embodiment, the motor is a Sorvall model RC5, the rotational speed is 300 to 22,000 rev / min, and the centrifugal force is 55,000 G. Further details of the manifold assembly 204 and adapter plate assembly 214 and their attachment are shown in FIGS. 5A and 6. In general, any suitable mechanism for driving the rotor and transmitting angular velocity to the rotor can be used. For example, a system using a compressed gas such as compressed air can be used to drive the rotor.

  As shown in FIG. 3, the dish assembly includes many columns such as 230. The chromatographic column has a cavity inside to be filled with a substance used for performing a chromatographic process. During operation, the internal cavity provides a conduit for the flow path (see, eg, FIG. 5A). When the chromatographic material is packed, chromatographic separation occurs along some channels in a column such as 230, for example.

  In the embodiment shown in FIG. 3, 24 columns are arranged around the dish-like assembly 202. As shown, in some embodiments, columns such as 230 may have a common length, a common outer diameter, and a common inner diameter. Columns can be made from common materials such as metals and alloys. Further, the columns are arranged around the outer periphery of the dish-like assembly 202 so that the columns are equally spaced. In another embodiment, a flexible material such as soft plastic is used for the conduit used in the rotating assembly.

  In other embodiments, the number of columns on the pan assembly can be varied and can be greater or less than 24. In addition, column length, column profile, and column inner diameter can vary from column to column on a single dished assembly. Furthermore, the composition of the chromatographic substance packed in the column can be changed for each column. Moreover, the space between columns does not need to be the same, and the space between columns can change for every column. Furthermore, the substances forming the column can also be changed from column to column (for example, the first column can be made of ceramics and the second column can be made of alloy).

  In other embodiments, the rotating assembly may comprise a number of dish-like assemblies. The dish-like assemblies can be arranged in layers, such as one on top of another. In some embodiments, a transmission and / or mechanism is attached to the dish assembly, which can be connected to or disconnected from the rotating shaft and rotated at different speeds. The number of columns can be varied by the dish-like assembly. In addition, column parameters such as length, outer shape, inner diameter, material and packing can be varied from one dished assembly to another.

  In the embodiment shown in FIG. 3, a flow cell such as 232 is installed at the end of each column. The flow cell 232 has a window portion and an internal cavity. The internal cavity serves as a flow path for fluid that elutes the chromatographic process from a column or the like. Light passes through the flow cell window.

  In one embodiment, the flow cells located at the end of each column can be common in similar designs, such as having windows at the top and bottom of the flow cells and having the same size interior. In another embodiment, the flow cell associated with a column can vary from column to column. For example, the flow cell associated with the first column on the pan assembly has windows on the side, whereas the flow cell associated with the second column has windows on the top and bottom. May be. Further, when the column parameters such as the column diameter are different for each column, the flow cell parameters such as the size inside the pipe line can be changed.

  In other embodiments, a flow cell need not necessarily be associated with each column. For example, the first column is associated with a flow cell, but the second flow cell is associated with a mass analyzer instead of a flow cell. In another embodiment, a flow cell and mass analyzer are associated with the column. That is, in general, the devices associated with the column for the carmatograph may be different for each column on one dish-like assembly, or may be different for each dish-like assembly. Further details of the apparatus used for the column performing the chromatographic processing are described in the “Apparatus” section below.

  As shown in FIGS. 2 and 3, a column such as 230 is secured in place by a column fastener 216. Column fasteners 216 are attached to the dished assembly by a plurality of fasteners, such as 236. In one embodiment, each column is installed using a retention mechanism, such as a preload nut 234. The preload nut 234 is visible through the column fastener 216. The preload nut can be secured to the pan assembly.

  During rotation, the column is subjected to large centrifugal forces such as hundreds to thousands G. As shown in FIG. 3, when the flow cell is located at the end of the column at the outer end of the dish-like assembly 202, the centrifugal force experienced by the column is transmitted to the flow cell, such as 232 for example. If too much force is transmitted to the flow cell, the shape of the flow cell, such as the shape of the flow cell window 242, may be distorted. Deformation of the flow cell, such as window distortion, degrades the optical properties of the flow cell and consequently degrades measurements using the flow cell. A load support mechanism such as a preload nut reduces the load on the flow cell such as 232 by the column during rotation. A preload nut associated with each column is mechanically secured to the pan assembly 202 to allow a portion of the load on the column to escape to a pan assembly or other support other than the flow cell.

  The liquid introduction assembly including the mixing chamber 228 is located at the center 238 a of the rotating assembly 200. In one embodiment, liquid enters the mixing chamber 228 from a stationary liquid reservoir while the rotating assembly 200 is rotating. For example, one or more conduits are inserted into the mixing chamber 228, and the mixing chamber 228 rotates while the conduits remain fixed. During operation, the fluid exits the conduit and enters the mixing chamber 228 to provide a continuously controlled flow to the rotating assembly 200.

  An assembly, including a conduit, that provides an interface to the rotor assembly 200 supports the conduit. The rotor assembly 200 includes a seat 240 for the gas bearing. The gas bearing is at rest on the seat 240 while the rotor assembly rotates. Further details of a gas bearing support assembly including a stationary liquid reservoir and conduit support are shown in FIGS. 7A-7C.

  The fluid enters the mixing chamber 228 and flows along the flow path from the center 238a of the pan assembly 202 to its distal end 238b. In one embodiment, the column is positioned along a radial line (ie, a straight line through the center of the dish assembly 202) and the fluid flows along one of the radial lines. In another embodiment, one or more columns are arranged along a non-radial straight line that does not pass through the center of the dish assembly. Nevertheless, centrifugal force components along a non-radial straight line can drive a flow through a non-radially arranged column.

  In some embodiments, fluid components from one or more reservoirs enter the mixing chamber 228 and are mixed. Mixing occurs by rotation of the mixing pin in the mixing chamber. The mixed liquid component exits the mixing chamber 228 through a number of ports, such as the ports associated with each column. Thus, in certain embodiments, two or more columns can receive fluid from a common reservoir. Further details of the mixing chamber 228 are shown in FIG. 5A.

  The fluid mixture passes through each column and enters the flow cell near the end 238b of the pan assembly 202. After passing through the flow cell, the fluid enters the return segment link 220 beyond the distal end 238 b of the pan assembly 202. The return segment link 220 includes a flow channel that routes the fluid exiting the flow cell along the dished assembly 202. A return segment link such as 220 is connected to the return channel next to the column. Once the flow enters the return channel, it leaves the end 238b toward the center 238.

  In this embodiment, fluid enters from the column tip located in the center of the dish-like assembly 202 and exits from the column end located outside the dish-like assembly 202. The column contains a packing such that the components are chromatographically separated in the fluid mixture as the stream travels from the top of the column to the end. As the fluid in the column moves toward the end of the dish assembly 202, the centrifugal force of the fluid in the column increases. Due to such orientation, as the chromatographic process proceeds, the centrifugal force increases and the pressure increases along the length of the column. The pressure gradient along such a column is different from chromatographic systems such as high performance liquid chromatography (HPLC). In HPLC, pressure drives the fluid from the tip to the end of the chromatographic column. Thus, the pressure decreases rather than increases from the top to the end of the column.

  In one embodiment, the maximum operating pressure of the rotor assembly reaches approximately 100 PSI. For example, when the pressure increases from the center of the rotor to the end of the rotor, the maximum pressure is 100 PSI or higher. In another embodiment, the maximum operating pressure is 50 PSI or higher. Maximum pressure levels, such as 100 PSI or less, are much less than the pressure of 1000 PSI for HPLC using smaller particle sizes.

  In one embodiment, return segment links such as 220 are connected to each other via connectors such as 241. The inner surface of each return segment link matches the outer surface of the dish-like assembly 202. For example, if the outer surface of the dish-shaped assembly 202 is curved, the return segment link matches the curvature of the outer surface of the dish-shaped assembly 202. The return segment link is secured to the dish assembly via a fastener such as 222. Further details of the return segment link are shown in FIGS. 5A-5C.

  After entering the return flow channel through the return segment link at the dish-like assembly 202, the flow travels to the circumferentially arranged channels at the disk portion 206 of the manifold assembly 204. Circumferentially arranged channels can drain fluid into a manifold block such as 225. Each manifold block is connected to an adjustable tube 226 that is connected to the column portion of the manifold assembly by a securing nut 224. The flow passes through the column portion of the manifold assembly and eventually enters the reservoir 210. Fluid is periodically drained from reservoir 210 via reservoir drain tubes 218a and 218b. Details such as the flow path through the interior of the rotor assembly 200 are shown in FIGS. 5A and 5B.

  4A and 4B are top and bottom views of the manifold assembly 204. FIG. There are circumferentially arranged drain channels 248 in the disk portion of the manifold assembly. An outer channel 242a and an inner channel 242b surround the drain channel 248. The outer channel 242a and the inner channel 242b support a circular gasket (not shown). When the top of the manifold assembly is connected to the bottom of the pan assembly 202, the circular gasket pushes the bottom of the pan assembly 202 and prevents fluid from leaking from the drain channel 248 during operation of the rotor assembly 202.

  Drain channel 248 includes a number of drain holes, such as 246. In FIG. 4A, six drain holes are shown. Each drain hole is connected to a manifold block such as 225. The manifold disk portion 206 has a number of holes, such as 251 for fasteners such as 254, which secure a manifold block such as 225 to the disk portion.

  An adjustable tube is connected to each manifold block 225. An adjustable tube end 250 can be inserted through the column 208 into a hole 252 in the column portion 208 of the manifold assembly and is connected to the central cavity portion of the column 208. A 250 like end of an adjustable tube such as 225 is found in the central drain column 244 of the manifold column 208.

  The manifold assembly 204 is connected to an adapter plate assembly 214 that includes an adapter plate (FIG. 6) through a locking hole, such as 258, in its bottom annulus 256. Fasteners are threaded through the mounting holes to secure the manifold assembly 204 to the adapter plate assembly 214. The bottom port 245 of the drain column 244 is connected to the drain channel of the adapter plate assembly 214 so that fluid exits the drain column 244 and enters the reservoir 210 through the adapter plate.

  As shown in FIG. 2, the reservoir 210 includes a number of drain tubes such as 218a and 218b. As previously mentioned, the reservoir 210 is periodically drained through the drain tube. In one embodiment, the drain tube passes through the bottom of the disk portion 206 of the manifold assembly 204 through a hole such as 255. The dish-like assembly has holes (not shown in the figure) through which drain tubes can be passed.

  As shown in FIG. 1, in the embodiment described herein, the flow branches and merges again. For example, in the rotor assembly 200, one flow path starts from the mixing chamber 228 and branches into 24 flow paths (see FIGS. 2 and 3). As shown in FIGS. 4A and 4B, the 24 flow paths merge into one drain channel 248. One drain channel 248 branches into six through a manifold block such as 225 and merges into a common drain column 244.

  In FIG. 4B, the diameter of the manifold assembly pan 206 is smaller than the diameter of the pan assembly 202. Since the diameter of the disk portion 206 is small, a window at the bottom of the flow cell such as 232a and 232b can be arranged. Thus, even if the light source is near the top of the flow cell, the light emitted from the light source, through the flow cell, and emitted via the bottom window can be collected. An apparatus platform for supplying light sources and collecting light emerging from the flow cell while the rotor assembly is rotating is shown in FIG.

  In FIG. 4B, the dish-like assembly 202 is partially transparent. As a result, a portion of the return channel tube associated with each flow cell is visible. For example, return flow channels 252a and 252b connected to flow cells such as 232a and 232b are visible. A fastener like 222 is also visible. A fastener such as 222 joins the return segment link to the dish-like assembly 202.

  In one embodiment, the return flow channel is formed by perforating a portion of the dish-like assembly 202. As already mentioned, the return flow channel, such as 252a or 252b, allows fluid to exit the flow cell and enter the return segment link to drain channels 248 arranged circumferentially at the disk portion of the manifold assembly 204. Moving away from the end of the dish-like assembly 202.

  The return flow channel is located beside each flow cell (not below), so that access to the bottom window of the flow cell is possible. In another embodiment incorporating different devices, the return flow channel takes different paths. Further details of the return flow channel are shown in FIG. 5B.

  FIG. 5A is a cross-sectional side view of the rotor assembly 200 showing the flow path. The flow path begins with a solvent reservoir 285 that allows one or more solvents to flow through the flow path. In one embodiment, many solvents are fed into the flow path simultaneously, where the fraction of the solvent composition can be changed over time. In this embodiment, the solvent reservoir is not on the rotor assembly 200. As already mentioned, in other embodiments, one or more solvent reservoirs can be placed on the rotor assembly 200. For example, the solvent reservoir can be connected to a column such as 230a or 230b of the dish-like assembly 202. One or more solvents are supplied by a pump (not shown). An example of a solvent reservoir on a rotating assembly is shown in detail in FIGS. 10A-10D.

  Returning to FIG. 5A, as already described in FIGS. 2 and 3, the gas bearing support assembly includes one or more flow paths (see FIGS. 7A-7C and 8 for details of the gas bearing support assembly 400). The one or more channels may be part of a channel such as 260. The flow path of the gas bearing support assembly 400 is used, for example, to supply solvent to the mixing chamber 228. In some embodiments, the flow path of the gas bearing support assembly 400 is also used to supply the sample used in the chromatographic process to the mixing chamber 228. In doing so, the sample is mixed with one or more solvents in the mixing chamber 228.

  In the mixing chamber 228, many mixing pins, such as 275, mix one or more fluids entering the mixing chamber. The mixing pin rotates according to the angular speed of the rotor assembly 200. At typical angular velocities, the mixing pin can remove air from the fluid. Thus, even when one fluid is fed into the mixing chamber and no fluid mixture is formed, the mixing chamber 228 has the advantage that air can be removed before the fluid enters a column such as 230a. .

  The mixing chamber 228 has a number of holes, such as 276, through which fluid can be drained from the mixing chamber. In one embodiment, each hole is in fluid communication with a chromatographic column containing one chromatographic stationary phase. In other embodiments, a single hole, such as 276, in the mixing chamber 228 may be fluidly connected to multiple chromatographic columns. For example, in the case of a tube-in-tube type, a large number of tubes may be provided inside the hollow cylinder, and each tube may contain a separate chromatographic stationary phase. The inlet of each tube is provided near the upper end of the hollow cylinder and is fluidly connected to one of the holes in the mixing chamber. During operation, fluid exits the mixing chamber and enters each tube via a hole. The cross section of the tube may be circular or non-circular.

  In another embodiment, the valve is set by the hole. The valve can control the rate at which fluid enters the hole. For example, a valve can be used to control the opening size of each hole. In some cases, the valve can be closed to prevent fluid from exiting the hole. For example, if a leak is detected downstream of a hole, the valve in that hole can be moved so that no more fluid exits through the hole.

  Many columns, such as 230 a and 230 b, extend from the mixing chamber to the end of the dish assembly 202. In one embodiment, the columns are arranged in symmetrical pairs to balance the rotor assembly 200 as it rotates. For example, columns 230a and 230b are a pair of symmetrical columns having similar masses. However, as already mentioned, the mass properties of the column pairs may be different for each column pair.

  In certain embodiments, the column may extend along a line perpendicular to the axis of rotation 265 that passes through the center of the rotor assembly 200. In other embodiments, the columns extend along a line that is not perpendicular to the axis of rotation (eg, each column can define two angles to the axis of rotation, such as the two angles used in a spherical coordinate system). But you can. For example, each column may be tilted downward or upward from the central axis 265 to the end of the dish-like assembly 202. To support a column that extends above or below the pan assembly 202, the pan assembly 202 may be thickened, or the pan assembly 202 may include additional support structures.

  As another example, instead of one column 230a extending toward the end of the rotor, the dish-like assembly 202 has two columns in this position, one tilted downward and the other tilted upward. May be connected to the mixing chamber and connected to the end of the dish-like assembly. In yet another example, the dish assembly 202 comprises three columns, the column 230a is straight across the dish assembly, the second column is tilted upward above the column 230a, and the third column is You may make it incline below 230a. Each column may be connected to the mixing chamber 228 or connected to a separate fluid reservoir.

  In yet another embodiment, the interior through which fluid flows in the flow path between the mixing chamber 228 and the dish-like assembly 202 is constant. For example, the inside of the column 230a and the flow cell 232 is approximately constant. In one example, the interior of the flow path, such as flow cell 232 or column 230a, is constant, but the shape of the periphery of the inner surface can vary along the flow path. For example, the internal shape of the flow path can be changed from a square with a certain area to a circle with the same area. At this time, when the shape changes between the two, the area is constant.

  In another embodiment, the interior of the flow path between the mixing chamber and the dish assembly 202 can vary between the mixing chamber and the end of the dish assembly 202. For example, a flow resistance that is narrower in the interior than other parts of the flow path can be placed at the end of the column 230a or behind the flow cell 232 to reduce the flow rate.

  During operation, rotation of the rotor assembly 200 can cause fluid to scoop up the sidewall 266 of the mixing chamber 228. In some embodiments, some of the sidewalls are parallel to the rotational axis of the rotating assembly, while other portions can be angled with respect to the rotational axis. It is also possible to change the shape of the mixing chamber. For example, the lower part of the mixing chamber 228 can be curved to form a bowl shape.

  As fluid is added to the mixing chamber 228 while the rotating assembly is rotating, a thickness of liquid head 268 is formed on the sidewall of the mixing chamber 228. The body head boundary is a free boundary that extends to some extent (eg, fluid head thickness) within the top, bottom, sidewalls, and mixing chamber 228 of the mixing chamber. The free boundary can approximate a cylindrical wall parallel to the axis of rotation of the rotating assembly.

  As shown in FIG. 3, the top of the mixing chamber is partially covered but has a hole. In some embodiments, the holes are circular, but other shapes are possible. As mentioned above, one or more flow paths pass through the holes so that fluid can be fed into the mixing chamber 228. As described in detail in FIGS. 7A to 7C, the upper portion of the mixing chamber may be covered by a gas bearing.

  The mixing chamber is sealed with a gas bearing to prevent release of the fluid contained in the mixing chamber 228 and its vapor. The gas bearing is kept stationary while the rotor assembly 200 including the mixing chamber 228 rotates. Thus, in operation, a portion of the rotating component and a portion of the non-rotating component form an enclosure that seals the mixing chamber 228. In another embodiment, an enclosure is formed that seals between two components rotating at different speeds. For example, a gas bearing located at the top of the mixing chamber may relate to a component that remains stationary or may relate to a component that rotates at a different speed than the rotor assembly.

  Also, fluid enclosures with fixed and non-rotating components or components rotating at different speeds are not limited to mixing chambers. For example, the mixing pins can be removed from the mixing chamber to form a fluid enclosure used as a fluid reservoir. Similar enclosures can be used to drain fluid from the rotor assembly. In this example, the bottom of the rotor assembly 200 may be placed on a gas bearing. The gas bearing may include a chamber covered by the rotor assembly 200 in a steady state. A channel that rotates with the rotor assembly 200 may extend from the rotor assembly 200 into the chamber to supply fluid into the chamber while the rotor assembly 200 is rotating. It is also possible to remove fluid from this chamber.

  Further details are shown in FIGS. 5A, 5B, 5C, 7A, 7B and 7C. The fluid flow is confined along a flow path formed on the rotor assembly from the mixing chamber 228 to a predetermined location, such as the reservoir 210, or via an interface that creates a flow out of the rotor assembly 200. . One advantage of confining fluid in various enclosures along the path through the rotor assembly is safety. Many fluids utilized in the devices described herein can be dangerous. For example, one fluid may be flammable and another fluid may be harmful to health, such as being carcinogenic. It can be operated more safely by confining the fluid inside the rotor assembly 200 along the flow path as compared to an open type interface in which the fluid and its vapor are easily released from the rotor assembly 200 (the fluid used in the rotor is If safe, the rotor assembly can be designed to include an interface that does not completely contain the fluid, such as an open interface where the fluid moves through at least a partially unsealed space).

  Returning to FIG. 5A, the rotor assembly 200 can be set such that the thickness of the fluid head in the mixing chamber can be adjusted. For example, the thickness of the fluid head can be adjusted by changing the flow path arrangement of the rotor assembly 200, and the fluid head does not extend until it enters the inner periphery of the upper hole in the mixing chamber. As a method of adjusting the thickness of the fluid head 268, first, a flow path that passes through the mixing chamber 228 and is connected to the outlet 250 of the adjustable tube 226 will be described. Second, an equilibrium condition is described so that the forces applied to the fluid in the two parts of the flow path are balanced. Finally, the setting adjustment of the rotor assembly 200 that affects the equilibrium conditions, including the fluid head thickness, will be discussed. Under equilibrium conditions, the thickness of the fluid head in the mixing chamber is a specific value. By changing the equilibrium conditions, the thickness of the fluid head in the mixing chamber 228 can be changed.

  In the rotor assembly 200, a straight line 262 is drawn at a constant radius 264 from the central axis 265 of the rotor assembly. The flow path is shown by the central axis 265 of the rotor assembly. The straight line 262 intersects the flow path through the rotor assembly 200 in the mixing chamber 228 near the outlet 250 of the adjustable tube 226. The outlet 250 can drain fluid from the adjustable tube to the central drain column 244 of the manifold assembly 204.

  When force is transmitted from the motor (not shown) to the rotor assembly 200, the rotor assembly rotates at an angular velocity about its central axis 265. As previously mentioned, the centrifugal force increases as the distance from the center axis of rotation 265 increases and as the angular velocity of the rotor assembly increases. This centrifugal force moves the fluid through the rotor assembly 200.

  For discussion purposes, two segments are defined in the flow path through the rotor assembly 200. The first flow path segment begins with the fluid wall on the free interface of the fluid head 268 in the mixing chamber 228 and moves outward (increases in radius) to the end in the return link. The second channel segment begins at the outlet 250 and moves outward (increases in radius) to the end within the channel segment link 220. The first flow path segment and the second flow path segment merge at the flow path segment link 220.

  In one embodiment, the rotor may be spun up from a resting state to a constant angular velocity to initialize the flow in the two flow path segments. Fluid introduction begins during the spin-up process (before reaching the target angular velocity) or after the rotor assembly 200 reaches the target angular velocity. In some embodiments, residual fluid from previous measurements may remain in the rotor assembly. The residual fluid may be contained in all or part of the first flow path segment and the second flow path segment. Therefore, another part may be dried by the initialization process in a state where the residual fluid is contained in a part of the first flow path segment and the second flow path segment.

  During the spin-up process, fluid introduction can begin in the mixing chamber 228. Not all rotor assemblies 200 must include a mixing chamber 228. In an embodiment, instead of using a common inlet such as 228, the rotor assembly 200 may include multiple fluid inlets, such as separate inlets for each column. Thus, this example is merely illustrative.

  Typically, in a chromatographic separation process performed in a column such as 230a or 230b, fluid passes from one end of the column to another. In the embodiment shown in FIG. 5, fluid is introduced from the head of the column closest to the mixing chamber 228 and passes to the tail 280. Many channels can be set to flow to the head of the column 270. For example, in the flow path shown in FIG. 5A, the flow begins near the center of the rotor in the mixing chamber and continuously moves from the center along the path to the column head. In another embodiment, the flow begins at a radius greater than the radius of the column head, moves toward the center of the rotor until it reaches the column inlet, and enters the column inlet.

  In another embodiment, a chromatographic column such as 230a or 230b can be installed on the return segment. Initially, the flow moves from the center 265 of the rotor assembly 200. At a predetermined radial distance from the center of the rotor, it turns to the center of the rotor and enters the return segment. A chromatographic column is installed where the flow moves in the direction of the center of the rotor, ie in the return segment. For example, the flow path can be set so that the flow moves from the end of the column 230a toward the head 270 of the column. In this example, a device such as a flow cell or a mass analyzer is installed on the head 270 of the column 230a.

  In another embodiment, the chromatographic column need not be linear. The column may be bent in any way from the rotor center to the rotor end. In general, the flow path following the path in the rotor assembly 200 is used to perform a chromatographic operation. The chromatographic flow path may be straight or bent. The chromatographic flow path may flow toward the center of the rotor assembly or may flow away from the center of the rotor assembly. Flows with different radial distances from the center of the rotor assembly, such as increasing, decreasing, constant or a combination of radial distances along the flow path before reaching the starting point of the chromatographic flow path where the flow enters. It flows along the road. Further, a flow that is longer or shorter than the radial distance of the portion where the flow flows into the chromatographic flow path may move in the radial direction. After reaching the end of the chromatographic flow path and discharging, the radial distance from the center of the rotor assembly is different, such as increasing, decreasing, constant or a combination of radial distances along the flow path. It may flow along the road.

  A reverse flow can also be set while operating the rotor assembly 200 in various modes. For example, when the flow on the return segment from the end of the rotor assembly 200 toward the center 265 is blocked by the action of the valve at the outlet, at a position that is radially away from the valve by a distance that is the position of the valve, A reverse flow toward the end of 202 begins and travels through the flow cell 232 and column 230a toward the center 265 of the pan assembly 202. Thus, in some embodiments, the flow path can be set so that the flow is in one direction at one point and the opposite direction at another point.

  Returning to FIG. 5A, during flow initialization, fluid introduced into the mixing chamber 228 exits the mixing chamber and enters a column such as 230a or 230b. Each column has two detachable caps such as 272 and 274 in the column 230b. In one embodiment, the two removable caps are threaded and secured to the end of each column. You may make it form a flow path in each cap. In one embodiment, the removable cap channel has the same cross-sectional area as the column.

  The first removable cap 272 provides an interface for connecting the column to a liquid introduction assembly that includes a mixing chamber 228. In some embodiments, the first removable cap includes, for example, a seat or groove for installing certain gaskets. The gasket is used to seal between the mixing chamber assembly and the first removable cap. Similarly, a second removable cap such as 274 provides an interface between each column and a flow cell such as 232. The second removable cap has an interface with a flow cell, such as a protrusion, inserted into the flow cell 232 in addition to one or more sealing seats or grooves, such as a gasket.

  If fluid is first introduced into the mixing chamber during the spin-up process, the angular velocity of the rotor assembly must exceed the threshold before fluid enters the column 230 or the like. If the angular velocity exceeds the threshold, the fluid in the mixing chamber 228 enters the column 230a and travels from the column head 270 toward the tail 280, forming a flow path 278 in the column 230a. The threshold of the angular velocity depends on the properties of the chromatographic packing material, such as the size of the packing material used for the chromatographic column 230. Typically, smaller diameter packed particles require greater RCF to move the flow compared to larger diameter packed particles. Since RCF is proportional to the square root of the angular velocity, a larger RCF can be produced by increasing the angular velocity of the rotor assembly.

  After the flow path 278 is created in the column 230 a, the fluid exits the column and forms a flow path 286 to the flow cell 232. The flow cell 232 includes an optical window, such as 282, and a hole that allows access to the optical window. The flow then exits flow cell 232 and forms flow path 288 in return segment link 220. As the flow proceeds through the return segment link 220, the first flow path segment from the mixing chamber 228 to the return segment link 220 is filled with fluid and the flow begins to enter the second flow path segment. The flow goes from the return segment link 220 to the center of the rotating assembly 200 and consequently reaches the outlet 250 of the adjustable tube 226.

  In the second flow path segment, the flow is directed from the end of the dish assembly 202 toward the central axis 265 and exits the return segment link 220 to form a flow path 290 in the return channel (in the return segment link 220 and the return channel). The details of the flow path are shown in FIGS. 5B and 5C). The flow exits the return flow channel, fills the drain channel 248, and forms a flow path 292 in the drain channel. As shown in FIG. 4A, the flow path 292 receives the flow of many return flow channels associated with different columns. Two gaskets are used to prevent fluid from leaking from the drain channel 248. A cross-sectional view of a gasket such as 275a or 275b is shown in FIG. 5A.

  In other embodiments, the second flow path segment only receives fluid from the first flow path segment. For example, the second flow path segment only accepts fluid from the flow through column 230a and flow cell 232, and does not accept fluid from other columns. In this embodiment, another return path (not shown) may be used. For example, one manifold block 225 and one adjustable tube 226 may be connected to one return channel from each column such as 230a, and the drain channel 248 may be omitted. As previously mentioned, the flow from multiple columns can be merged in the drain channel to reduce the amount of flow path used in the rotor assembly 200. Manufacturing costs can be reduced by reducing the amount of flow paths.

  After the flow path enters the drain channel 248, it exits through one or more holes into one of the manifold blocks, such as 225, to form a flow path 294 in the manifold block 225. The manifold block 225 is connected to the adjustable tube 226. The flow exits the manifold block 225 and forms a flow path 295 in the adjustable tube 226. The flow path in the adjustable tube 226 ends at the outlet 250.

  Fluid is introduced into the mixing chamber 228 until the thickness of the fluid head reaches the same radial distance 264 of the outlet 250. At this point, if no more fluid is introduced into the mixing chamber, the fluid head thickness 268 reaches equilibrium at approximately the same location as the outlet 250 so as to satisfy Bernoulli's incompression equation. Due to viscosity and other non-linear effects such as the rotor assembly precessing relative to the central axis 265, the radial position of the outlet 250 and the thickness of the fluid head are not exactly the same, but approximately At the same distance. If more fluid is introduced into the mixing chamber 228, the fluid head thickness reaches a radial position that is less than the outlet 250. If the fluid head thickness 268 is greater than the equilibrium position, the flow in the rotor assembly 200 will attempt to return to equilibrium. As a result, the flow moves past the rotor assembly 200 in a direction out of the mixing chamber 228. Excess fluid spills out of outlet 250.

  When the introduction of fluid into the mixing chamber stops at some point, fluid spills out of the outlet 250 and enters the drain column until the fluid head thickness 268 is approximately in the radial position with the outlet 250. When fluid is continuously introduced into the mixing chamber drain column 244, the flow enters the drain column 244 via an adjustable tube and forms a flow path 296 in the drain column. The flow exits the drain column 244, passes through the bottom of the manifold assembly 204, and eventually reaches the interior of a reservoir such as 210a. Details after the flow exits the manifold assembly 204 are shown in FIG.

  The radial position of the fluid interface associated with the fluid head and outlet 250 can vary. In the embodiment of FIG. 5A, during rotation, a flow is created against the centrifugal force by the centrifugal force during rotation and is adjusted from the bottom of the adjustable tube across each outlet, such as 250, from the bottom of the mixing chamber to the top of the mixing chamber. Flows to the top of the tube. As a result of surface tension and gravity, the fluid head interface from the top to the bottom of the mixing chamber or the fluid interface across the outlet 250 is curved and the radial distance from the central axis of rotation 265 varies with the location of the fluid interface. . As the angular rotation speed of the rotor assembly increases, the radial change along the liquid interface decreases, i.e., the interface becomes more straight in the vertical direction.

  In one embodiment, when equilibrium is reached such that the radial position of the fluid head thickness 268 is approximately the same as the outlet (ie, the fluid head thickness 268 is approximately the same as the 262 line). The rotor assembly 200 increases the rotational speed to a constant angular velocity (as described above, fluid can also be introduced into the mixing chamber while the angular rotational speed of the rotor assembly increases with time. ). After the rotational speed of the rotor assembly 200 reaches a predetermined target angular velocity, fluid is added to the mixing chamber 228 and the fluid passes through the rotor assembly 200 until it reaches the outlet 250 and begins to flow out of the adjustable tube 226. Flowing. By adjusting the rate at which fluid is added to the mixing chamber 228, the flow rate in the rotor assembly 200, including the flow rate in columns such as 230a and 230b, can be controlled.

  As described above with reference to FIG. 1, after determining that the steady flow rate has been reached in the columns such as 230 a and 230 b, the sample is introduced into a predetermined position of the rotor assembly 200. For example, a sample is added to the mixing chamber 228. From this mixing chamber, the sample is introduced into each column. In another embodiment, shown in detail in FIG. 9A, the sample is injected near the head of a column such as 230a or 230b. If the column contains a chromatographic filler, the sample is chromatographed and analyzed by measurement using a flow cell 232 at the end of each column.

  The fluid head equilibrium thickness 268 can be adjusted by changing the distance 264 from the axis of rotation to the outlet 250. For example, the fluid head is adjusted to the equilibrium thickness of the fluid head so that the fluid head does not enter the hole in the seat 240 of the mixing chamber 228 during operation. As already noted, the holes in the mixing chamber seat 240 provide an inlet for one or more flow paths and can supply fluid to the mixing chamber 228.

  In general, in various embodiments, the rotor assembly 200 includes: 1) In the first flow path segment, the flow is along a flow path and the radial distance from the center is at the end of the flow path Then it is larger than the initial distance. 2) In the second flow path segment connected to the first flow path segment, the radial distance from the center is smaller than the initial distance at the end of the flow path. An outlet is placed at the final radial distance of the second flow path segment, and the fluid exits the second flow path segment. The distance from the rotation axis 265 where the outlet is located in the second flow path segment determines the equilibrium position of the first flow path segment. Fluid is introduced into the first flow path segment at a rate. The speed at which fluid is introduced into the first flow path segment is used to control the flow rate through the first flow path segment and the second flow path segment. As described below, the fluid introduction speed and the associated flow rate are relatively insensitive to changes in the angular rotation speed of the rotor assembly 200.

  Returning to FIG. 5, in another embodiment, the radius 264 of the outlet 250 of each adjustable tube 226 can be adjusted. An outlet, such as 250 for each adjustable tube, can be installed at the same or different distance from the central axis 265 of the rotor assembly 200. An adjustable tube such as 226 has a groove that fits the manifold assembly 204 and a groove that fits the manifold block 225. The adjustable tube is screwed into the manifold block 225 at various depths and the radial distance of the outlet 250 changes. The adjustable tube 226 is fixed by the fixing nut 224 so that the radial position of the outlet 250 is constant during the rotation of the rotor assembly 200.

  In one embodiment, the rotor assembly 200 has a mechanism for dynamically changing the radial distance of the outlet 250. For example, an elastically extensible flexible end part is attached to the end of an adjustable tube such as 226. During operation, force is applied to the flexible terminal part, changing the radial distance of the outlet 250 connecting to that part. For example, when no force is applied to the flexible end piece, the outlet 250 remains near the wall of the drain column 244, but when the force is applied, the outlet 250 extends in the direction of the center of the drain column 244.

  The advantage of setting the rotor assembly 200 with the first flow path segment to the end of the rotor connected to the second flow path segment is that once the equilibrium condition is established, the first flow path is established via the mixing chamber 228, for example. The flow rate can be controlled by changing the speed of introduction into the segment. As ideal conditions are approached, the rate of fluid introduction into the first flow path segment required to obtain a constant flow rate in the column is essentially independent of angular velocity at wide angular velocity operating conditions.

  Furthermore, the flow rate in the mixing chamber 228 required to obtain a specific flow rate in the rotor assembly 200 is less dependent on angular velocity compared to other flow settings returning from the end of the rotor. For example, the flow exits from the rotor end, such as the return segment link 220, rather than 250. In one embodiment, a fluid recovery ring, such as 210, surrounds the rotor end so that, for example, fluid exiting the return segment link may be recovered (in this example, the return segment link causes flow to flow to the center of the rotor assembly. 265). Compared to a configuration in which the flow is diverted back to the rotating shaft 265 as shown in FIG. 5A, in this configuration, the outflow velocity at the rotor end and the velocity through the column are far greater than the change in angular velocity. Greatly affected. Typically, for this type of configuration, as the angular velocity of the rotor assembly increases, the RCF also increases, so the outflow rate also increases.

  5B and 5C show further details of the flow near the rotor end. In particular, FIG. 5B shows a cross section of a dish-like assembly 202, a column 230 (also referred to as stationary phase material) that contains a packing material carried in the chromatographic process, a flow cell 232, a return segment link 220, and a return channel such as 298a and 298b. Is a view seen through from above, showing the flow path. The column 230 may be placed on the dish-like assembly 202. The dish-like assembly 202 may include a groove in which a column is accommodated.

  As described above, a removable cap 274 can be combined with the end of the column 300. In one embodiment, the removable cap 274 has a protrusion that is set to fit the opening of the flow cell 232. A gasket 274a surrounds the protrusion and maintains a seal between the removable cap 274 and the flow cell. The dish-like assembly 202 has a recess in which the flow cell 232 is placed.

  The flow cell 232 has an opening that allows fluid to flow toward the return segment link 220 (see 252a and 252b in FIG. 5C). The flow path exits return segment link 220 and enters return channels 298a and 298b. A gasket 220 a is attached between the return segment link 220 and the dish-like assembly 202. The gasket 220a prevents liquid leakage between the return segment link 220 and the flow cell 232 and between the return segment link 220 and the return channel.

  The dish-like assembly 202 has a receiving portion for the fastener 222a. The fastener is inserted into a hole in the return segment link 220. By tightening the fastener, the force applied to the gasket 220a between the return path segment link 220 and the dish-like assembly 202 can be increased. Due to the force applied to the gasket 220a, the sealing performance is increased and leakage can be prevented. The return flow channel has a portion 298a that enters the pan assembly 202 through the outer end of the pan assembly 202 and a portion 298b that goes down through the pan assembly 202 (298a and 298b are angled with respect to each other). ). The portion of 298b is connected to the drain channel 248.

  FIG. 5C shows an example of the return segment link 220. The return segment link 220 has holes such as 300 and 306 that can be combined with one or more return segment links. One return segment link is combined with another return segment link by passing a pin through a hole such as 300 or 306. When all the return segment links are combined together, a continuous chain ring around the rotor end is created.

  The return segment link 220 has a hole 304. As shown in FIG. 5B, the return segment link and dish assembly 202 are combined by inserting a fastener into the hole. In one embodiment, the return segment link 220 has two recesses, such as 302a and 302b. This recess allows flow between the flow cell on the pan assembly 202 and the return flow channel. In one embodiment, each path segment link 220 integrates flow from two different chromatographic columns.

  The inner surface of each return path segment link 220 may be curved. When the return segment links are connected, the inner surface forms a circle and the surface is curved so that it is slightly larger than the circle surrounding the periphery of the end of the dish-like assembly 202. The surface curve 308 distributes well the forces resulting from fasteners attached through holes 304. The distributed force on the surface 308 improves the seal between the return segment link 220 and the outer end of the pan assembly 202.

  FIG. 6 shows a side view of the rotor assembly 200, with the reservoir being a cross-sectional view and the adapter plate assembly 214 being a partially transparent view. Adapter plate assembly 214 includes an adapter plate 318. The manifold assembly 204 is combined with the adapter plate 318 via a fastener such as 320. A tapered shaft 212 connected to a motor (not shown) is combined with a disk portion 214a of the adapter plate assembly 214 via a fastener 316. The taper shaft 212 is inserted into the bottom of the adapter plate assembly 214 and fixed with a fastener.

  The reservoir 210 is combined with the disk portion 214a of the adapter plate assembly 214 and the like. The adapter plate 318 has one or more flow paths. This creates a flow path from the drain column 244 in the manifold assembly 204 to the adapter plate. One or more gaskets (not shown) are used to keep the fluid interface of the manifold assembly 204 between the liquid interface on the adapter plate 318.

  Fluid enters the adapter plate 318 from the manifold assembly 204, passes through one or more flow paths, and exits the adapter plate through one or more holes 310. In one embodiment, the flow enters from the center of the rotor assembly and flows radially toward the reservoir 210. Thereafter, it flows into the reservoir 210a and the like.

  The fluid interface between the reservoir 210 and the adapter plate 318 can be sealed. In one embodiment, there is a channel outside the adapter plate 318 that is sealed with gaskets 314a and 314b. The advantage of this method is that the adapter plate 318 can be inserted inside a single reservoir drum such as 210. The reservoir drum 210 is provided with a circular protrusion 322 or many arms extending from its inner surface. The adapter plate 318 is placed on this protrusion or arm. The disk portion 214a of the adapter plate assembly is inserted through a circular protrusion or arm. A circular protrusion or arm provides a place to attach the reservoir drum to the adapter plate 318.

  In some embodiments, the reservoir drum is divided into a plurality, such as two halves or four quarters. The adapter plate 318 has a flow path that is inserted into the hole of the reservoir drum 210, or the reservoir drum 210 has a flow path that can be inserted into the adapter plate. The reservoir drum 210 is assembled from a number of parts. Such a configuration replaces the sealed channel at the end of the adapter plate.

  Different rotor assemblies can employ different manifold assemblies and different reservoir designs. For example, the inner diameter of the drain reservoir drum 210 and the outer diameter of the manifold assembly 204 can be increased. Different adapter plates 318 and reservoir drums can be used for different configurations of the manifold assembly 204, with a portion of the adapter plate assembly connected to the tapered shaft 212 being reusable.

  The rotor assembly shown in FIGS. 5A, 5B, 5C and 6 is made up of a number of modular parts. The module configuration is only given as an example and is not limiting. The advantages of modular construction are assembly, disassembly, configuration flexibility and low cost. The disadvantage of modular construction is that as the number of parts in the module increases, the fluid interface increases and the need for sealing increases, resulting in an increased possibility of liquid leakage.

  In some embodiments, several parts may be integrally formed to perform functions related to two separate parts in order to eliminate potential leak locations. For example, in some embodiments, the drainage reservoir and manifold assembly may be integrally formed so that the flow path through the adapter plate 318 is omitted. In another embodiment, the mixing chamber enclosure may be formed as a separate component from the pan assembly 202. In yet another embodiment, the mixing chamber enclosure may be a component integrally formed with the pan assembly 202. Further, the manifold block 225 and the adjustable tube 226 can be formed as one flexible conduit.

  In another embodiment, there are many grooves on the dish-like assembly 202 on which the column can be placed. Thus, for example, the column can be removed or re-installed for packing or washing. In other embodiments, the column may be integrally formed with the dish assembly 202. For example, many holes can be drilled in a disk forming a dish-like assembly and used as a column. In another example, a channel can be created inside a dish-like assembly such as 202 by pouring metal onto the channel and later removing it.

  In another embodiment, a number of open channels may be formed on the first plate, after which the channels are covered. For example, the channel may be covered by adhering the second plate. Even after covering the channels, each channel has an inlet and an outlet. For example, many channels arranged in the radial direction may be formed on a first “washer” type disk. Each channel is formed from the inner radius to the outer radius of the washer. Thereafter, a second structure such as a second washer having a flat surface with the same dimensions as the first washer may be adhered to the first washer to form a closed channel. In one embodiment, two separate plates are formed with, for example, semi-cylindrical open channels. When the two plates are brought together, a closed channel with a circular cross-section is formed.

  7A-7C show a side view and a projected view of the gas bearing support assembly 400. FIG. The gas bearing support assembly 400 may be installed on a drum-type rotation mechanism (see FIG. 8) that rotates the rotor assembly. The lid 414 functions as a drum lid. The lid is made of a hard and transparent material such as bulletproof glass. If it is made of bulletproof glass, it can be operated while looking at the rotor assembly while protecting from parts that are blown from the rotor assembly due to some mechanical trouble. In other embodiments, the lid can be formed of a non-permeable material.

  A gas bearing 418 is installed on the seat 240 during rotation of the rotor assembly 200. The rotating rotor assembly 200 can be stabilized by the weight from the gas bearing 418 to the seat 240. For example, the weight of the gas bearing support assembly 400 reduces precession that occurs while the rotor assembly rotates.

  The gas bearing support assembly 400 includes a number of flow paths. The flow path 416 passes through the hollow shaft 412 and passes through a hole in the center 426 of the gas bearing 418. As previously mentioned, when the gas bearing 418 is above the seat 240, the flow path may extend to a chamber below the seat 240, such as a mixing chamber 228, for example. In addition, the bottom of the gas bearing 418 may serve as part of the lid of the mixing chamber to help confine fluid within the mixing chamber.

  In one embodiment, there are three channels through the gas bearing support assembly. The three channels can be used to supply various fluids to the mixing chamber, such as two different solvents and sample fluids. Gradients can be easily applied by changing the concentration of different solvents over time. In other embodiments, more or fewer channels pass through the shaft 412 to the mixing chamber. In some cases, the gas bearing support assembly does not include any flow paths and is used only to stabilize the rotor assembly. In other cases, one or more flow paths are inactive and not used.

  The shaft 412 has one or more fluid interfaces such that one or more flow paths from the solvent reservoir 285 and / or the sample injector 402 lead to a flow path through the central portion of the shaft. For example, the shaft 412 is connected to the cap 406 by an interface connected to a flow path by a flexible tube from the solvent reservoir 285 or the sample injector 402. One or more pumps associated with the solvent reservoir or sample injector 402 move the fluid into the shaft.

  In one embodiment, the one or more pumps are positive displacement pumps such as piston driven pumps. In another embodiment, the one or more pumps are positive displacement pumps such as buoyancy pumps. A compressed air driven double diaphragm pump is an example of a buoyancy pump. This type of pump runs on top of the compression pump and has the least operating part. In another embodiment, a gas amplification pump is used.

  The gas bearing 418 is porous. Gas can pass through the pores in the gas bearing. When the gas bearing 418 is on the seat 240 and the rotor assembly is rotating, the gas compressed through the gas bearing 418 forms a thin layer between the gas bearing 418 and the rotating seat 240. A thin layer of gas minimizes friction between the gas bearing 418 and the rotating seat 240. Sometimes the gas bearing and the rotating seat 240 are in contact. This contact results in the gas bearing 418 being lost.

  The gas bearing includes an interface to a gas source 404, such as a pressurized argon container. During operation, pressurized argon passes through the gas bearing 418. The advantage of argon is that it is the third most gas and inert in the atmosphere. However, other gases may be used, and argon is merely an example.

  The gas bearing support assembly 400 is shown with the shaft 412 and gas bearing gas bearing support assembly in the raised state in FIG. 7A and lowered in FIG. 7B. In some embodiments, the gas bearing support assembly is raised and lowered by lever 408. The lever 408 is connected to the rotating shaft 410, and the rotation 420 of the shaft 410 moves up and down the shaft 412 and the gas bearing 420. The rotating shaft 410 is supported by a support block 424. Support block 424 is combined with lid 414 by one or more fasteners, such as 428.

  FIG. 8 shows a side view of the rotor assembly 200, the gas bearing support assembly 400 in the containment structure, and device fixtures such as 444 and 442. FIG. The gas bearing support assembly 400 is mounted in place and a gas bearing 418 is located on the seat 240 of the rotor assembly 200. The gas bearing lid is on the ledge of the containment structure 440.

  In one embodiment, the containment structure 440 is drum-shaped and made of metal. The containment structure 440 has a hole, and the tapered shaft 212 is connected to the motor under the rotor assembly 200. The motor can transmit the angular velocity to the rotor assembly 200.

  One or more holes are provided in the storage structure or lid 414. The hole serves as a passage for exhausting gas from the chamber 450. In some embodiments, the chamber 450 is sealed and some degree of vacuum is achieved in the chamber. A gasket is placed between the lid 414 and the containment structure 440 to improve sealing. In addition, fasteners are used to more securely combine the lid 414 and the storage structure. One or more holes in the lid 414 or the containment structure 440 are connected to the vacuum pump. When the vacuum pump is activated, a certain degree of vacuum is achieved in the chamber 450. The vacuum pump is also used for safety purposes to prevent accidental generation of dangerous gases such as flammable gases in the chamber 450.

  One or more device fixing brackets such as 442 and 444 are provided in the storage structure. In some embodiments, each device fixture, such as 442 and 444, includes a light source and a light collector. The light source and the light concentrator may be arranged in line with the flow cell window on the rotor assembly 200 as described above. In order to place the light source and light concentrator in line with the flow cell window near the end of the rotor assembly 200, the device fixture may extend above or below the rotor assembly 200.

  In one embodiment, two fiber optic cables are attached to the device fixture. The first fiber optic cable is coupled to a light source that emits photons at one or more wavelengths. The second optical fiber cable is connected to the photomultiplier tube. The second fiber optic cable receives photons emitted from the first fiber optic cable that have passed through the flow cell. The collected photons are passed to a photomultiplier tube. The photomultiplier tube and the light source are located outside the storage structure 440.

  The signal from the photomultiplier tube can analyze the component of the fluid passing through the flow path. A light source coupled to each device emits light of a different wavelength. For example, there are visible light, ultraviolet light, infrared light, and the like, but it is not limited thereto. In various embodiments, a device capable of emitting and collecting light may not be used, or one or more such devices may be used.

  In other embodiments, device fixtures such as 442 and 444 may include gas bearings. The gas bearing stabilizes the rotor assembly during rotation and prevents the rotor assembly from colliding with equipment fixtures such as 442 and 444. A flow path such as a flexible tube can be coupled with the device fixture to supply gas to the gas bearing. The flow path is connected to a gas source for use with the gas bearing.

  Device fixtures such as 442 and 444 include revolute joints that are partially rotatable. By using a rotating joint, a portion of the device fixture, for example, the upper part thereof, can be rotated, eg, rotated upward, so that the rotor assembly can be placed within the containment structure 440 and the containment structure. It is also possible to arrange outside 440.

  In one embodiment, if there is sufficient clearance between the side of the containment structure and the end of the rotor assembly, a portion of the device fixture, such as the part containing the fiber optic cable, is placed on the retractable arm. Is possible. By retracting the retractable arm, the rotor assembly can be installed and removed from the retractable structure. When the retractable arm is extended, optical fiber cables and other devices on the device fixture attached to the retractable arm can be placed above and / or below the rotor assembly.

  As the rotor assembly rotates, many rotor parts rotate repeatedly past the device fixture. An indexing system may be provided to keep track of the current position of the rotor, such as the position of the rotor where the measurement is taking place. This indexing system can uniquely identify where the measurement is taking place on the rotor assembly. The created index may be associated with the measurement using the device installed in the device fixture.

  As an example, the rotor assembly 200 may be provided with an identifier such as a marking that can identify the position of the rotor. For example, a visible marking may be provided on the dish-like assembly of the rotor assembly 200 to uniquely identify the position such as the position where each column is arranged. The marking may be, for example, a number and / or a symbol such as a character or a barcode. In another example, the RFID tag storing the identifier may be arranged at various positions around the end of the rotor assembly 200, for example, at the position where each column is installed.

  A detector may be placed on the device fixture to detect when the identifier passes through the device fixture. For example, you may make it detect identifiers, such as a symbol and a barcode, using a camera. As another example, an identifier such as a barcode may be read using a laser and a detector. As another example, an RFID tag reader may be used to receive an identifier from an RFID tag installed on a dish-like assembly.

  In operation, many measurements are possible. Measurements can also be performed on individual chromatographic columns. The amount of measurement performed depends on the angular velocity of the rotor assembly and the length of time that the measurement is made. The detected identifier may be used as an index to map a set of measurements to predetermined features of the rotor assembly, such as individual columns and associated flow cells. Indexed measurements can be used to create time-varying profiles of measurements for individual chromatographic columns.

  FIG. 9A is a top view of an embodiment of a rotor assembly used in a chromatographic system. The parts of the rotor assembly were previously shown in FIG. The power generation ring 333 can be combined with the rotor assembly. The power generation ring 333 includes power generation components such as one or more wire coils and a transformer circuit.

  When used as a generator, the power generation ring 333 passes through a magnet such as a magnet placed on the apparatus fixture shown in FIG. As the coil passes through the magnet, current is induced in the one or more coils. The generated electricity can be supplied to devices on the rotor assembly. In another embodiment, the power generation ring 333 may include a number of batteries and be arranged in a balanced and even manner around the rotor assembly. The battery may or may not be rechargeable by a generator located on the rotor assembly.

  In another embodiment, a brush interface is used to supply power to the rotor assembly 200. For example, a thin conductive tape is installed near the end of the storage structure 440 shown in FIG. One or more arms with one or more conductive brushes or other contact mechanisms are extended from the rotor assembly 200 and contact the conductive tape to which the conductive brush is secured to supply power to the rotor assembly 200. To do. Since the conductive brush is insulated, no power is supplied to the entire apparatus. In another embodiment, a portion of the rotor assembly 200, such as the tapered shaft 212, includes an insulated conductive tape. The fixed conductive tape is located within the encapsulating structure 440 and contacts the insulated conductive tape on the rotor assembly. During rotation, electricity is transferred between the metal brush and the rotor assembly, driving various devices on the rotor assembly.

  Electricity from outside the rotor and power generated on the rotor can be adjusted by a transformer circuit so that it can be used in various devices on the rotor assembly 200. One or more conductors, such as 334, for example, take leads from the power generation ring 333 and supply power to various devices. The regulated electricity charges the battery on the rotor and moves environmental conditioning devices such as electrically driven valves, communication devices, devices on the rotor, sensors on the rotor, and heating elements.

  In FIG. 9A, the conductor 334 is coupled to the heating elements 329 and 331 and the sample introduction mechanism 325. The heating element rings such as 329 and 331 may include one heating coil, or may include a plurality of heating elements to which electricity is supplied from the power generation ring 333. You may make it provide a control apparatus in any one or both of a heating element ring. In this case, both rings may be controlled by one control device, or each ring may be controlled by a separate control device. The heat output of each heating element ring may be controlled by one or more control devices. In one embodiment, either or both heating element rings may comprise one or more heating elements for each column, and control the heating elements of each column independently. A temperature sensor may be installed along the column, and the target temperature may be controlled based on data received from the temperature sensor, for example.

  In one embodiment, the sample introduction mechanism 325 may be installed on the rotor. For example, the sample introduction mechanism 325 includes a fluid interface such as an electronically operated valve, and the sample reservoir of the sample introduction mechanism is connected to each column so that the sample stored in the sample reservoir is injected into the column. May be. The sample introduction mechanism 325 may include one or more pump-type injection devices on the board and inject the fluid stored in the sample reservoir into each column. For example, the injection device may be a syringe pump.

  In addition, the sample introduction mechanism has one or more replenishment ports, such as 327, each connected to a sample reservoir. The sample reservoir can be replenished at the refill port. In some embodiments, samples can be supplied to multiple columns from a single reservoir. For example, a single replenishment port such as 327 is connected to a single sample reservoir from which it is fed to two or more columns. Furthermore, the sample in each sample reservoir can vary from reservoir to reservoir. Thus, during operation, many different samples are chromatographed simultaneously on the rotor. In other embodiments, separately controlled injectors can be used when multiple sample reservoirs are used. The injectors are controlled separately and different samples are introduced at different times. The sample introduction mechanism 325 may include a control device that controls a device such as a sample injection device or a controllable valve disposed on the sample introduction mechanism.

  The rotor may include a built-in electric circuit 337 such as a processor, a memory, a battery, and / or a communication interface. In one embodiment, the electrical circuit 337 includes a wireless communication interface to allow communication between the rotor and the system management device (shown in FIG. 1). The electric circuit 337 may include a communication bus, and data may be transmitted from the electric circuit 337 to many devices. For example, the electric circuit 337 receives a command from a remote device, and transmits the command to a specific device on the rotor assembly, for example, the power generation ring 333, the heating element ring 329 or 331, the sample introduction mechanism 325, or the adjusting valve. But you can.

  Many devices that generate data on the board are coupled to the communication bus 341 via a wire-like communication link, such as 339. In other embodiments, wireless communication links are used and used for communication between many devices on the rotor. The advantage of a wireless link is that the wire path through the rotor assembly can be minimized. For example, many devices are wireless transmitter / receivers and can communicate with the electrical circuit 337.

  Data generated by the device is sent to the electrical circuit 337 via wireless and / or wired communication. The electrical circuit 337 stores the data and / or sends it to the remote device. The rotor may have many sensors. Examples include, but are not limited to, a temperature sensor, a flow rate sensor, a water level sensor (for example, the reservoir 210 and the mixing chamber 228 have a water level sensor), a pressure sensor, and combinations thereof. Data from the sensor is communicated to the remote device via electrical circuit 337. In other embodiments, a device has its own communication interface that is not connected to the electrical circuit 337. For example, sample introduction device 325 is configured to communicate directly with a remote device without going through an intermediate device such as electrical circuit 337.

  As another example, the rotor may have a device on the board that generates measurement data. The device on the board communicates with the electrical circuit 337 to send data to the remote device. In one embodiment, data is sent in real time as it occurs. In another embodiment, one or more devices include a memory to store data and later send it to a remote device. Since the memory also serves as backup data, even if real-time data is lost due to malfunction of the transceiver, for example, it can be compensated from the memory.

  FIG. 9B shows a side view of a rotor assembly in an embodiment configured for use in a chromatographic system that includes a fraction collection mechanism 350. Generally, the fraction collection mechanism receives eluent eluted from the chromatograph. Therefore, the fraction collection mechanism is considered as an eluent reservoir. The position of this fraction collection mechanism is merely illustrative. If the rotor configuration is different, the column and flow path will be different, and the position of the fraction collection mechanism will also change.

  In some embodiments, a fraction collection mechanism may be coupled to a flow path located in the return flow channel on the dish-like assembly 202 by a fluid interface, such as a valve 352. As shown in FIGS. 5A and 5B, the return flow channel is coupled to the exit of a flow cell such as 232 via a return segment link 220. The valve 352 is opened and closed at various timings, and the fluid discharged from the flow cell 232 is sent to the fraction collection mechanism 350 or the fraction collection mechanism 350 is bypassed. Valve 352 may be opened and closed while the rotor assembly is rotating.

  In one embodiment, a command to open and close valve 352 is received from a device outside the rotor. Based on information such as measurements made in the flow cell, the remote device determines when to open and close the valve and sends a command to open and close the valve. In another embodiment, a controller on the rotor assembly makes this determination and sends an open / close command to valve 352.

  The fraction collection mechanism 350 may have one or more chambers for storing collected fractions. For example, the fraction collection mechanism 350 has four chambers having four valves 354 and can be controlled to individually flow fluids. A chamber such as 356 has a passage hole, and the collected fraction can be taken out. For example, in one embodiment, fractions can be collected from the holes when operation is terminated and the rotor assembly is at rest.

  One or more columns are attached to the fraction collection mechanism 350. With a single fraction collection mechanism, fractions can be taken from a single chromatographic column or from multiple columns. The number of fraction mechanisms on the rotor assembly will depend on the mapping between each fraction collection mechanism and the number of each column as well as the total number of columns on the rotor. Furthermore, when a fraction collection mechanism is used, it is not necessarily associated with every column on the rotor. Thus, many fraction collection mechanisms on the rotor assembly vary depending on the form of the rotor assembly.

  In yet another embodiment, a variable volume eluent collector such as 350 can be placed behind a detector such as a flow cell. There is an electric motor that can wirelessly control the volume-variable eluent collector, and the lead screw is rotated inside a cylindrical chamber. A piston-like hermetic washer is mounted on the lead screw. When the lead screw rotates, the piston-like hermetic washer moves toward the outside of the chamber, and the liquid flows at a speed that varies depending on the moving speed of the piston-like hermetic washer.

  The variable volume eluent collector can be incorporated into a reservoir such as 210 as previously described. In another embodiment, a device such as a reverse syringe pump can be used as the variable volume eluent collector. In some cases, it is necessary to place a variable volume eluent collector with multiple chambers at the end of each column for fractionation. In such a case, a multiport valve may be added between the column and the variable volume eluent collector.

  FIG. 9C is a block diagram that includes a reaction chamber located between the end of a column such as 230 and an apparatus involved in detection such as a flow cell. In some cases, after chromatographic separation, a chemical reaction is performed on the analyte exiting column 230. As a result of the reaction, the analyte can be more easily identified with a particular detector. For example, if the analyte is non-fluorescent, it cannot be detected with a fluorescence detector such as, for example, LIF (see instrument section below). By chemical reaction, it can be converted into a substance that has the property of easily detecting an analyte and can be detected by a specific detector.

  In one embodiment, a flow diverter such as 800 switches the flow from the column and flows directly to the detector, such as 232, or to a reaction chamber such as 802 before the flow cell. Reaction chamber 802 has an access port to which one or more different reagents such as 804 and 806 can be added alone. The access port is controlled by a valve such as 808 or 810. In the example shown here, all or part of the column is contained in a reaction chamber such as 802. In addition, a reaction chamber, such as 802, receives analytes from one or more different columns.

  The reaction chamber may include an ultrasonic oscillator. In some embodiments, ultrasonic vibrations can be generated using piezoelectronic elements. Ultrasonic vibration can increase the reaction rate of the analyte and complete the chemical reaction. In some embodiments, a reaction chamber 802 can be installed between the column 230 and the flow cell without using a flow diverter such as 800. In this embodiment, reagents can be added to the reaction chamber as needed or passed without adding reagents. In yet another example, reaction chambers are arranged in series to perform multistage reactions.

  In other examples, another energy source may be used to affect the reaction and increase the reaction rate in the reaction chamber 802. For example, the reaction chamber may be irradiated with microwaves using a microwave generator. In another example, a temperature control mechanism may be used to heat or cool the reaction chamber as needed.

  10A to 10D show another system for performing a hanging bucket type centrifugal liquid chromatography. FIG. 10A shows a front view and a side view of the bucket assembly. Includes multiple columns for chromatographic separation. The bucket assembly is removable. In operation, the bucket assembly 500 is rotated in combination with the rotor assembly, as shown in FIGS. 10B-10D. The rotation applies a centrifugal force to the bucket assembly. The bucket assembly is combined with the rotor assembly by a rotor support 506. During rotation, fluid in many chromatographic columns flows due to centrifugal force and chromatographic separation occurs.

  The bucket assembly 500 has a solvent reservoir in which a solvent and sample mixture are previously placed. Solvent reservoir 502 is combined with many chromatographic columns via well plate 504. During rotation, solvent and sample pass through the openings in well plate 504 by centrifugation. In one embodiment, the well plate has 96 openings and is combined into 96 columns.

  In another embodiment, each bucket assembly is combined with a central reservoir of a rotor assembly that rotates together. The central reservoir is located near the axis of rotation of the rotor assembly (see FIG. 10C). The central reservoir controls the fluid mixture such as solvent composition and sample for the elution gradient supplied to each rotor assembly. In one embodiment, a flexible fluid interface, such as a flexible hose, is attached from the central reservoir to the solvent reservoir 502 on each bucket, and fluid from the central reservoir is supplied to the solvent reservoir 502. As will be shown below, the position of the bucket assembly changes from a vertical position to a horizontal position during the spin increase. The flexible fluid interface is used to accept changes in the position of the bucket assembly during increased rotation.

  A pack 510 including a battery and a transceiver can be coupled to the bucket assembly 500. Electric power can be supplied to the flow rate control device 514 using a battery. In one embodiment, the flow rate control device 514 may be an electric stop cock. The electric stop cock can be connected to each column. When the stop cock is closed, no liquid can pass through the column 508. When the stop cock is partially opened, liquid can pass through the column 508. As the liquid begins to descend through the column, air displaced from the column is released through a vent, such as 516.

  You may make it influence the flow velocity in each column using the position of a water stop cock. When the stopcock is fully open, the flow rate increases through the column. In one embodiment, the flow rate control device 514, such as a stopcock, can be remotely controlled. The remote device sends a command and sends a request to a specific stopcock via a transceiver. Data related to the flow rate controller 514, such as the current state, is sent to the remote device via the transceiver.

  In certain embodiments, the sample reservoir is located above the solvent reservoir 502 so that the sample is initially stored away from the solvent. A mechanism such as a valve can be used to introduce the sample into the solvent reservoir. During operation, the solvent first travels through the column and forms a stream in the column. The sample is then introduced into a solvent reservoir, such as 502, and the solvent and sample mixture travels down column 508. The solvent reservoir 502 has a mixing mechanism such as an ultrasonic mixer, and mixes the solvent and the sample after introducing the sample.

  Many flow cells 512 can be installed near the end of column 508. The flow cell is used to make optical measurements on the flow leaving column 508. Details of the optical measurements that can be performed are shown in FIGS. 10B-10D. After exiting the flow cell, the waste liquid is collected in solvent reservoir 518.

  FIG. 10B is a top plan view of the rotor assembly 525 and includes a plurality of buckets such as 500. In this view, the rotor assembly 525 and bucket are rotating. Rotor assembly 525 rotates about axis 501. The rotor assembly 525 has a fastener such as 520 for attaching a bucket such as 500. In the embodiment of FIG. 10B, the rotor assembly 525 has four fasteners for attaching four buckets, such as 500, as needed. The rotor assembly 525 is surrounded by a container 534 such as a metal drum.

  In the stop state, buckets such as 500 are arranged in a straight line in the vertical direction by the gravity vector (see FIG. 10C). During operation of the rotor assembly 525, its rotational speed increases, so the bucket 500 rotates to the horizontal position and lifts as shown in FIG. 10B. In the horizontal position, the transceiver on the bucket passes the transceiver position 524 located at the bottom of the container 534. Via the transceiver location 524, data originating from each bucket is sent to the remote device, or data or instructions are sent from the remote device to each bucket. For example, a command is sent to a flow rate control device disposed on each bucket.

  In another embodiment, many 500-like buckets are attached to a dish-like assembly as shown in FIGS. 2-8 so that the buckets are always in a horizontal position. When using a dish-like assembly, the bucket is connected to a fluid reservoir, such as a mixing chamber 228, for supplying solvent and sample, and is connected to a further flow path so that waste liquid from the bucket is transferred to a reservoir, such as 210. To enter. Furthermore, since the bucket is incorporated in the flow path, the flow rate determined by the fluid introduction speed is substantially independent of the angular velocity. That is, the flow returns from the center of the pan assembly to the end and back toward the center.

  Two instrument assemblies are shown. Each instrument assembly is attached to a fixture 520, such as 530, to a retractable arm 528 with one or more sensing elements such as a retractable arm 528, 526 associated with the fixture. Things are included. While arm 528 is in the extended position, arm 532 is in the stowed position. When arm 528 is extended, detector 526 is on line 522 of the optical flow cell and measurements can be made using the flow cell. When arm 532 is in the stowed state, the device associated with the instrument assembly is not used.

  In some embodiments, multiple instrument assemblies can be utilized. However, not all instrument assemblies and devices are used for all operations of the chromatographic system. When the instrument assembly is not utilized, it can be stowed as shown in FIG. 10B.

  FIG. 10C is a top plan view of a rotor assembly that includes a plurality of buckets in a resting state. FIG. 10D is a top plan view of a rotor assembly including a plurality of buckets in a rotating state. The rotor assembly has a central reservoir 570. The central reservoir can control the fluid composition for each basket. Fluid from the central reservoir 570 is supplied to the bucket via the flexible fluid interface 572. The flexible fluid interface 572 has a detachable connector 574 that is connected to a receptacle on the bucket.

  In certain embodiments, the central reservoir 570 may include multiple chambers for storing different mobile phase fluids. Further, the reservoir may include a gradient forming part and a mixing chamber. The gradient forming unit controls the composition of the mobile phase fluid using different fluids. Further, the central reservoir may comprise a sample introduction mechanism for introducing a sample and one or more sample reservoirs. The sample may be mixed with the mobile phase fluid in a mixing chamber. The mixing chamber may be fluidly connected to each bucket via a flexible fluid interface such as 572.

  In some embodiments, a flexible power interface (not shown) such as a power ship can be combined with a flexible fluid interface. A flexible power interface is attached to a receptacle on the bucket to provide power to a device such as a control valve on the bucket. The flexible power interface is combined in the center of the rotor assembly. As shown in FIG. 9B, during rotation, a brush interface is used to supply power to the rotating element from a power source located other than the rotating element.

  A brush interface is used to provide power to the central portion of the rotor assembly during rotation. Thus, the bucket also receives power from a power source outside the rotor via a flexible power interface between the central portion of the rotor assembly and each bucket. The power source outside the rotor is also used to move the device associated with the central reservoir 572. In another embodiment, a power source on the rotor, such as a generator or storage battery, moves the central reservoir 572 and device on the bucket.

  In the example shown in FIGS. 10C and 10D, there are seven fixtures 550, 552, 554, 556, 558, 560 and 562 for seven different types of detectors. All or some of the detectors can be used during operation. In the example of FIG. 10D, all of the detectors are available. Accordingly, all arms involved in the detector are shown in use in FIG. 10D. In this case, for each rotation of the rotor assembly, seven different measurements are made in each flow cell and repeated for each subsequent rotation.

  In certain embodiments, detector 550 may be a visible and ultraviolet absorption detector. The detector 552 may be a fluorescence detector. Detectors 554, 556, 558, 560 and 562 are laser induced fluorescence detectors (LIF). Each LIF detector uses a different wavelength laser. Details of devices such as absorption detectors, fluorescence detectors, and LIFs are described in the next section.

One or more detectors are used to identify the eluent eluted from the column after the sample has been chromatographed, as is done while rotating one of the instrument columns. In chromatography, the fact that the retention time in the column matches the retention time of a known compound is not necessarily sufficient information for identifying the eluate. Therefore, an additional detector is required.

  In chromatography, the retention time exhibited by a substance under specific chromatographic parameters and the response characteristics to a specific detector are the basis for chemically identifying the substance. Since all substances respond differently to each detector, the more detectors used, the more certain the chemical identification of the substance. If these parameters are combined in addition to the retention time for one or more reference substances, chemical identification can be ensured.

  A detector is a system that outputs a response when an eluting substance is detected and gives a peak signal on a chromatogram. Typically, a detector is placed immediately after the stationary phase to detect when the compound elutes from the column. For example, as mentioned above, flow cells are used for many optical detections. Thus, the flow cell is typically located immediately after the column.

  The information generated by the detector can be displayed as a time variation value, for example, as a curve having peaks and valleys that change with time when different sample components are eluted from the column. The coarse and detailed adjustments associated with the detector adjust the peak width and height. Also, the sensitivity parameter associated with the detector is often controlled.

  As previously mentioned, in certain embodiments, the detector and associated elements are located on a rotating element, such as a rotor assembly. In addition, the signal associated with the detector is also processed on the rotor assembly. For example, the flow cell, associated detector, and processor for performing signal processing are located on the rotor assembly shown in FIGS. 2-9C or the bucket assembly shown in FIGS. 10A-D. In other embodiments, a portion of the detector is placed on the rotor assembly and the remaining portion is secured outside the rotor, such as the flow cell or detector shown in FIGS. 2-10D. In yet another embodiment, the detector is located completely outside the rotor. For example, the collected fraction is transported out of the rotating element by some mechanism for further analysis.

  There are many types of detectors available in the embodiment shown. The detectors available in the examples shown here are: refractive index (RI), ultraviolet (UV), visible (VIS), fluorescence, radioactivity, electrochemical, near infrared, mass spectrometry ( MS), nuclear magnetic resonance (NMR), light scattering (LS) and combinations thereof. These detectors are shown below.

  There are two types of detectors in liquid chromatography. The first type requires the detector transducer to be in direct contact with the effluent. There are various types of electrochemical detectors and mass spectrometers. The second type does not require that the physical elements of the detector be in direct contact with the effluent. This includes not only nuclear magnetic resonance detectors, but also devices that measure the electromagnetic spectrum at some or several locations.

  A refractive index (RI) detector is used as a measure of the ability of an analyte to bend and reflect light. This property of each molecule or compound is called the refractive index. In some types of refractive index detectors, especially differential refractive detectors, light travels through a two-module flow cell to the photodetector. One channel of the flow cell passes the mobile phase through the column and the other channel passes only the mobile phase. When light is bent by the sample eluting from the column, detection is done by reading the difference between the two channels. In other types of RI detectors, the degree to which light is refracted as the analyte passes through the flow cell is directly detected without reference to another flow cell.

Ultraviolet (UV) and visible (Vis) detectors are used to measure the ability of a sample to absorb light. This measurement is made at one or several wavelengths. Typical UV detector sensitivity is approximately ^10-8 or ^10-9 gm / ml. A UV detector usually requires a light source and a light collection element. Usually, a lamp having a wide wavelength range, a light-emitting line lamp, a light-emitting diode, or a laser is used as the light source. As previously mentioned, the light emitted from the light source passes through the flow cell, but is collected in the transmissive window of the flow cell using several focal points or filter elements.

  As an example, the absorption at a short wavelength such as 254 nm is measured. It is possible to measure at different wavelengths by tunable measurement. At the same time, it is a single wavelength but covers a wide range of wavelengths. For example, in an embodiment where the flow cell rotates under a fixed detector, the first wavelength of the light source is the first round of the rotor assembly, the second wavelength is the second round, and in the third week. Again, the first wavelength is used. This type of measurement can be repeated for many two or more wavelengths. In another embodiment, a diode array is used to simultaneously measure the spectrum versus wavelength. In yet another embodiment, several different light sources, with or without concentrating elements, are mounted in a fixed position relative to the rotating element and focused on the flow cell, with each rotation of the rotating element on the rotating element Measure at different wavelengths using one common flow cell.

  Using the scanning UV / Vis detector, the spectrum of the entire visible ultraviolet (UV-Vis) of the sample can be measured and detected. This instrument is very effective for the identification and analysis of sample compounds. This type of detector measures and records the response of the material passing through the flow cell to light. In order to detect a wide wavelength range, the detector proceeds in one of two ways. In the first method, the entire wavelength range is scanned by a scanning monochromator. Standard scanning monochromators use tungsten or deuterium lamps that emit light in a wide wavelength range. Light travels from the grating or prism through the slit to the sample cell. The sample is detected by a photomultiplier tube. The wavelength is adjusted by rotating the grating or prism, but one location is scanned at a time. The data for each data point is subsequently obtained at a different time.

  In the second method, all UV-Vis regions are measured simultaneously. In one method, a plurality of photomultiplier tubes are installed and used at a position where a wavelength region of interest can be detected. Another method uses a linear photodiode array (LPDA) spectrophotometer to measure the 190-1100 nm region simultaneously. Monochromatic light is not required and data is acquired in milliseconds. With this detection method, kinetic intermediates can be analyzed, and overlapping peaks can be separated and analyzed using spectrometry.

A fluorescence detector measures the ability of a compound to absorb light at one (excitation) wavelength and emit light at a slightly longer wavelength. Each compound may have a characteristic fluorescence. Excitation light reaches the photodetector through the flow cell, while a monochromator measures the emission wavelength. The sensitivity of a typical fluorescence detector is approximately 10 ^-9 to 10 ^-11 gm / ml.

  Laser induced fluorescence (LIF) is a spectroscopic technique that can be used to detect chemical species. As noted above, the embodiments described herein use one or more LIF detectors. The chemical species of interest are excited by monochromatic light from the laser. The wavelength is selected so that the chemical species has the largest absorption cross section. The excited chemical species is usually de-excited after a certain time in the nanosecond to microsecond range, and emits light having a wavelength longer than the excitation wavelength. The emitted light and fluorescence can be measured with some detector.

  In some embodiments, emission and / or excitation spectra are measured. In order to measure the emission spectrum, the laser wavelength is fixed and the generated fluorescence spectrum is measured. In order to measure the excitation spectrum, the emission wavelength is fixed, the wavelength of the laser is changed, and the fluorescence is measured.

  One advantage of a fluorescence detector is that two or dimensional images can be obtained because fluorescence occurs in all directions (ie, the fluorescence signal is isotropic). Furthermore, the fluorescence signal is highly sensitive because of its high S / N ratio. Many chemical species can be distinguished by adjusting the excitation wavelength to a wavelength unique to the target chemical species so as not to overlap with other substances.

Radiochemical detection is a method using a radioactively labeled substance such as tritium ³ H or carbon-14 ( ¹⁴ C). Fluorescence associated with ionization of beta particles is detected. One application is the study of metabolites. There are two types of detection: uniform type and non-uniform type. In a homogeneous detector, scintillation fluid is mixed with column eluent and fluorescence is generated. In the heterogeneous detector, the fluorescence from the beta particle emission and lithium oxalate interact with the detection cell. The typical detection limit for this type of detector is 10 ^-9 to 10 ^-10 gm / ml.

Electrochemical detectors are used to measure compounds that undergo oxidation or reduction reactions. Usually, the moving sample is measured by measuring the receipt or loss of electrons as it passes through an electrode between a certain potential difference. In some embodiments, electrodes are placed near the ends of one or more columns in an arrangement such as the rotor assembly or bucket assembly described above. The typical detection sensitivity of this type of detector is ^10-12 to ^10-13 gm / ml.

In mass spectrometry (MS) detectors, sample compounds or molecules are ionized. Thereafter, the ion current is detected through the mass analyzer. There are various ionization methods. In the first method, often referred to as electron impact (EI), the sample eluted from the column is ionized by a current or beam created in a high electric field. In the second method, often referred to as chemical ionization, an ionized gas is used to move electrons from the compound eluting from the column. In the third method, often referred to as fast electron impact, fast xenon atoms are used to ionize the elution chamber from the column. The detection limit of this type of detector is 10 ⁻⁸ to 10 ⁻¹⁰ gm / ml. In one embodiment, the method is performed on the rotating element during the rotation of the element.

In a nuclear magnetic resonance detector, certain nuclei having an odd number of masses, such as H and ¹³ C, randomly rotate around the axis. However, when placed in a strong magnetic field, the spins are aligned parallel or antiparallel to the magnetic field, and the parallel orientation is slightly more energetically stable. When the nucleus is irradiated with electromagnetic waves, it absorbs it and changes from parallel to high energy state and resonates. H or C shows a different spectrum depending on the position in the molecule and the adjacent molecule or element. This is because all the nuclei in the molecule are covered with an electron cloud, which changes with the surrounding magnetic field and has a different resonance frequency.

  In a light scattering (LS) detector, some light is reflected, absorbed, transmitted, or scattered as the light source emits a collimated beam that strikes particles in the solution. In the turbidimetric method, light scattered by a solution containing particles is measured, but a part of the light scattered at a certain angle is detected. Since detection is performed in the absence of background, the sensitivity depends on background light.

  Turbidity measurement is a method of measuring the decrease in light transmitted by particles in a solution, but measures light scattering as a decrease in light passing through a solution containing particles. Therefore, the amount of remaining light is measured. The sensitivity of this method depends on the sensitivity of the equipment used, but ranges from simple spectrophotometers to high-end dedicated analyzers. Thus, the reduction in transmitted light signal is limited by the accuracy of the photometer and the equipment used.

  Near-infrared detectors work by scanning compounds in a wavelength range such as 700 to 1100 nm. The stretching and bending vibration of chemical bonds in each molecule can be detected at a specific wavelength. In this detector, a plurality of analyzes can be performed from one spectrum.

Chromatographic separation As mentioned above, chromatography is the process of physically separating the individual components of a chemical mixture. In chromatographic processing, a mixture of chemicals is dissolved in a carrier stream (gas or liquid). A carrier stream containing the mixture is passed through the particle bed. The carrier stream moves through the particle bed at a certain speed. In chromatography, the carrier stream is called the “mobile phase” and the particle bed is called the “stationary phase”.

  In the examples described above, a chromatographic enclosure including a stationary phase such as a cylindrical column has been described. The chromatographic enclosure contains a chromatographic stationary phase. At least one flow path passes through the chromatographic enclosure, and fluid enters the chromatographic enclosure, passes through the chromatographic stationary phase, and exits the chromatographic enclosure.

  In one embodiment, many separate flow paths are provided in the chromatographic enclosure and there is no fluid connection between the separate flow paths. A separate chromatographic stationary phase is included in each channel. Separate flow channels allow parallel chromatographic processing, with each sample channel processing a common sample fluid or separate sample fluids in parallel.

  The flow path is secured by a structure including an internal cavity. The fluid and chromatographic stationary phase are contained within the structure containing the internal cavity. G Generally, fluid is driven axially through a structure containing its internal cavity. For example, a thin pipe is formed, and the fluid passes through the pipe in the axial direction. The length, inner cross section, area thereof, etc. are defined as the structure of the cavity portion associated with each flow path in the chromatographic enclosure. In some embodiments, the inner cross-section is constant along the length of the flow path. In other embodiments, the inner cross section varies along the length of the flow path. In another embodiment, the inner cross section is circular. In general, the inner cross section may have any shape.

  In some cases, the outer cross section of the structure may be similar to the inner cross section of the cavity portion. For example, the inner cross section and the outer cross section are circles with different diameters to form a cylinder. For example, a square block can be made by drilling a right-angle block. In one example, the chromatographic enclosure may be integrated with the rotor, or may be integrated with a case separate from the rotor.

  Each chromatographic enclosure has an inlet and an outlet. The inlet allows fluid to enter one or more flow paths. The outlet allows fluid to exit from one or more flow paths. A fluid, such as a chromatographic mobile phase, enters from the inlet, passes through the chromatographic stationary phase, and exits from the outlet to establish a fluid flow path to the chromatographic enclosure.

  The chromatographic enclosure is operated by a rotor that supplies centrifugal force to the chromatographic enclosure. For example, as described above, the chromatographic enclosure is operated on a rotor assembly that is adjusted to rotate at an angular rotational speed. Centrifugal force forces the fluid to pass through the chromatographic enclosure. The fluid includes a sample. When the fluid containing the sample passes through the stationary phase contained in the chromatographic enclosure, chromatographic separation of the sample occurs.

  As an example, FIG. 11 shows a front view of a chromatographic enclosure for chromatographic processing before and after centrifugation. A column such as a cylindrical plastic syringe 600 is held in a container such as a centrifuge tube 604. The cylindrical plastic syringe 600 has an upper opening 615 and a lower opening 616. In this example, since the upper opening 615 is larger than the lower opening 616, the cross-sectional area of the column changes in the length direction. As mentioned earlier, in other examples, the cross-sectional area of the column is constant in the length direction.

  Many elements are added to the plastic syringe 600 prior to centrifugation to set up the chromatographic process. A multi-hole filling such as glass wool 616 is added near the bottom of the syringe 600. The multi-hole packing allows fluid to exit the column without a stationary phase such as 610 exiting the column.

  In one embodiment, the stationary phase is a chromatographic adsorbent 610. The type of stationary phase used varies depending on the type of chromatographic treatment that is set. The sample interacts to varying degrees with the stationary phase material, depending on the type of solvent used and the type of stationary phase material. The types of chromatographic processing and the characteristics of the stationary phase and solvent used are shown below in line with the description of FIG.

  A sample 608 dissolved in a liquid is placed on the stationary phase. The size of the stationary phase particles is selected so that the sample does not enter the stationary phase by gravity alone. A porous separation medium such as sand 606 is placed on the sample 608. A separator separates the sample from the solvent, such as solvent reservoir 602. The solvent serves as the mobile phase for chromatographic processing. It is placed on the sample and separator 606.

  The chromatographic processing arrangement is then combined with a device that provides angular velocity to one of the rotor assemblies previously described. Under centrifugal force, the solvent in solvent reservoir 602 and sample 608 are driven to a stationary phase, such as 610, and flow downstream through the column. Sample 608 and solvent in solvent reservoir 602 pass through the stationary phase during centrifugation. After some time, the sample 608 moves the stationary phase 610 by a distance. Thus, the stationary phase is shown above and below sample 610. A portion of the mobile phase passes completely through the stationary phase 610 and exits as the eluent 618, collecting the column outlet 616 at the bottom of the centrifuge tube 604.

  This example is merely illustrative and does not limit the present invention. In the example of FIG. 11, the sample begins to move through the stationary phase before a steady flow is formed in the column. Thus, chromatographic separation begins before a steady flow is formed. In the embodiment described above, a steady flow may be formed before sample introduction.

  In the example of FIG. 11, a solvent composition that is a mixture of different solvents is applied to the solvent reservoir 602. In the previously described example, during centrifugation, the solvent reservoir 602 is not replenished and the solvent composition does not change. In another example, the solvent reservoir 602 is replenished during centrifugation and the solvent composition changes, i.e., gradient elution occurs.

  Depending on the composition of the mobile phase and the composition of the stationary phase, different types of chromatographic processing are performed. The embodiments shown here are applicable to any type of chromatographic process in which the mobile phase passes through the stationary phase of the chromatographic enclosure. Examples of chromatographic processes using the methods and apparatus described herein include partition chromatography, adsorption (liquid-solid) chromatography, ion exchange chromatography, affinity chromatography, size exclusion chromatography such as gel permeation and gel filtration chromatography. There are graphics, but this is not the case. Partition chromatography generally includes chemically bonded and adsorbed phase reversed phase chromatography in addition to conventional partition chromatography.

  In adsorption chromatography, a liquid or gaseous mobile phase is used and adsorbs on the surface of the stationary phase. Separate solutes with different equilibrium differences between the mobile and stationary phases. Partition chromatography produces a thin coating on the stationary phase surface. The thin film is fixed to the surface of the stationary phase particle by a covalent bond (chemical bond type) or adsorbed on the surface of the stationary phase (non-bonded type). In ion exchange (IEX) chromatography, oppositely charged solute ions in a mobile phase liquid are attracted to the resin (or particulate stationary phase) by electrostatic attraction, and the higher the analyte charge, It takes a long time to interact strongly with the surface and pass through the chromatographic system. IEX chromatography is also effective in determining the tertiary and quaternary structure of the produced protein. This is because the separation is performed under the condition using only the aqueous solution.

  Size exclusion chromatography does not use the attractive force between the stationary phase and the solute. Instead, a liquid or gas passes through the porous gel and the molecules are separated according to the size of the gel. The pores are usually small and exclude large solute molecules, but small molecules enter the gel and flow through a larger volume. As a result, large molecules pass through the column at a faster rate compared to small molecules.

  Affinity chromatography takes advantage of the specific interaction of one solute molecule with another molecule immobilized on a stationary phase. For example, an antibody specific to a certain protein is immobilized. As the protein mixture passes by the molecule, the specific protein reacts with the antibody and binds to the stationary phase. This protein is later removed by changing the pH of the ionic strength.

  More particularly, reverse phase partition chromatography (RPC) includes all chromatographic techniques using non-polar stationary phases. Unlike forward partition chromatography (NPC), it has a stronger affinity for highly polar compounds on unmodified silica or alumina with a hydrophilic surface. In RPC, the elution order in the case of NPC is reversed by introducing an alkyl chain with a covalent bond to the stationary phase surface. In RPC, the parish compound elutes first and retains the nonpolar compound—thus called “reverse phase”.

  In RPC, an inert and non-polar substance can be used because it can sufficiently cover the surface of the packed particles. One example is commercially available on silica 297 (USP classification L1) with attached octadecyl groups. Another example is a C8 bonded silica column (L7-166 commercially available). In addition, there are a cyano group-bonded column (L10-73 is commercially available) and a phenyl group-bonded column (L11-72 is commercially available). C18, C8 and phenyl types are used for reverse phase fillers, while cyano types are used in reverse phase mode depending on the analyte and mobile phase conditions. It should be noted at this point that not all C18 columns necessarily show the same retention characteristics. When functionalizing the surface of silica, a monomeric or polymeric reaction is carried out using organosilanes of different alkyl chain lengths to suppress the remaining silanol groups in the second stage (end-capping). Although the overall retention mechanism is the same, the surface chemistry differs slightly for each stationary phase, so a difference appears in selectivity.

  In RPC, a mixture of water (or aqueous buffer) and significant solvent is used to elute the analyte from the reverse phase column. The solvent is miscible with water and is a common solvent such as acetonitrile, methanol, or tetrahydrofuran (THF). In addition, ethanol, 2-propanol (isopropyl alcohol), or the like is used. Elution is performed isocratic (the water-solvent composition does not change during the separation) or gradient (the water-solvent composition changes during the separation). The pH of the T mobile phase has an important effect on analyte retention and changes the selectivity of certain objects. Charged objects can also be separated on a reverse phase column using ion pair generation (called ionic interaction). This method is known as reverse phase ion pair production chromatography.

The efficiency of the comparative chromatographic separation process between the example results and HPLC is often referred to as “chromatographic efficiency”. Chromatographic efficiency theory provides a relative indicator of the efficiency of many chromatographic processes to be compared. If the method and apparatus shown here are used, the chromatographic process can set a more effective chromatographic process as compared to a similar chromatographic process performed using other methods and apparatuses. Thus, in the following paragraphs, methods and apparatus for centrifugal liquid chromatography, including pressure requirements and particle size limitations, are compared to HPLC, a form commonly used in liquid chromatography. In addition, a simple description of chromatographic separation theory is provided, specifically 1) a measure of the efficiency of chromatographic separation using the apparatus and method described herein, and 2) other types of chromatographic equipment such as HPLC. And a comparison with the performance measure obtained in.

Back pressure is an amount commonly used in HPLC. Back pressure uses a calculated back pressure to determine the pressure required to pass fluid through a column with a particular size particle as a stationary phase. The pressure required for HPLC is calculated as follows.
ΔP = (ηFL) / (K ⁰ πr ² d _P ² )
Where ≡P is the pressure at the head of the column and falls in the length direction, η is the viscosity, F is the flow velocity, L is the length of the column, K ⁰ is the intrinsic permeability, r is the radius of the column and d _P Is the particle diameter. Increasing the flow rate, fluid viscosity, column length, or decreasing particle size requires higher pressure resistance. For example, fine particles of 2 micrometers or less by HPLC require high pressure resistance. For example, when using 2 micrometer particles in HPLC, a pressure of 10,000 PSI or more is required.

  In this example using centrifugal liquid chromatography, the need for back pressure and the importance of pressure are different from HPLC. For example, at certain particle size sizes, as well as the other parameters shown above, the pressure at which the chromatographic system shown is operated is much less than that required for the HPLC system. Also, the system using centrifugal liquid chromatography behaves differently from the HPLC system with respect to pressure. For example, in the example shown here, the pressure increases along the length from the head of the column, but conversely decreases in HPLC.

  Without being bound by a particular theory, the column consists of many layers and the fluid descends from column to layer. In the separation efficiency theory, as described above, each of these layers is called a “stage”. For example, when pressure is used as a driving force as in HPLC, a slight pressure is required to flow a fluid across each layer. Thus, the required pressure depends on the layers or stages in the column, and the overall required pressure is the sum of the pressures for fluid to flow across each layer. Separation efficiency usually increases as the number of layers increases. Therefore, in HPLC, separation efficiency that increases as the number of layers increases requires higher pressure.

  In centrifugal liquid chromatography, there is no pressure loss because the fluid crosses each layer by centrifugal force. Since centrifugal forces act on each layer independently of each other, there is no pressure loss across the layers as in HPLC. Thus, centrifugal liquid chromatography has a much lower pressure requirement than HPLC. Applicants believe that this feature has not been fully appreciated in the prior art.

  Furthermore, the examples described here can use much finer particles than are possible with HPLC. For example, particles of approximately 15 angstroms can be used. If even smaller particles are produced, they are also considered usable.

  In chromatography, a plate model is used to evaluate the separation efficiency. The stage model considers many layers in a chromatographic column. This is called “theoretical stage”. It is assumed that the sample equilibrates between the stationary and mobile phases, but this occurs at each “stage”. The analyte is lowered from one stage to the next by the equilibrated mobile phase.

The term “stage” is used as an analogy to the process that functions in the column. There is actually no step in the column. The column efficiency, that is, the ability to perform chromatographic separation, is quantified as the number N of theoretical plates in the column, or a height comparable to one theoretical plate, the theoretical plate height. For a column of length L, the theoretical plate height is defined as
HETP = L / N.
Comparing the chromatographic processes that occur in two different columns, a column with a high N value or a low HETP value indicates high chromatographic efficiency.

Separation efficiency has been defined for liquid chromatography occurring in columns. The International Pure Applied Chemistry Union defined the separation efficiency as follows:
N = 16 (V _R / w _b ) ² = 16 (t _r / w) ²
Here, N is the number of theoretical plates, V _R is the volume of the mobile phase flowing into the column between the sample injection and the peak maximum appearance of the sample component, and w _b is completely from the time when the analyte starts to leave the column. The total volume of time to finish, t is the retention time (seconds), and w is the peak width (seconds).

Experimental measurements were made using the chromatographic system of the examples described herein. A column with a length of 2.3 cm and a void volume of 0.110 ml packed with 5 micron diameter (d _p ) particles was used for the separation of the three species. Chemical species 1 is FD & C RedNo. 3 (erythrosin). Chemical species 2 is FD & C YellowNo. 5 (Tatrazine). Chemical species 3 is FD & C GreenNo. 3 (First Green FCF, E143). The mobile phase is 70% water, 30% isopropyl alcohol (IPA) and 0.010 mol / liter tetrabutylammonium phosphate (TBAP). The theoretical plate height was obtained in this experiment. These experiments showed the values shown in Table 1.

The values of V _R and w _b for each species shown in relation to formula of theoretical plates. k ′ is the retention factor of each species, α is the ratio of the retention volumes of the two species, R is the resolution between the two species, N is the number of theoretical plates, N / L is the number of theoretical plates, and the column length (0 .023m). For example, the degree of separation between chemical species 1 and 2 is calculated as follows. In general, R (A, B) = 2 [V _RB− V _RA ] / [w _bB + w _bA ]. For 1 and 2, R (1,2) = [V _R-2- V _R-1 ] / [w _b-2 + w _b-1 ]. α (A, B) = k ′ _A / k ′ _B is the ratio of the retention coefficients of the two chemical species. For example, α (1,2) = k ′ ₁ / k ′ ₂ .

  The chromatograms of the three chemical species shown in Table 1 are shown in FIG. The amount shown in the table such as the theoretical plate was calculated from the data obtained in the chromatogram such as the volume of each peak. Although it looks like a line on a small scale, the peak is wide and used to calculate the volume.

  The degree of separation between the theoretical stage and the chemical species for each chemical species is extremely high. For example, a degree of separation between two chemical species of 1.5 or more is considered complete separation. The theoretical plate number and resolution are much higher than the theoretical plate number and resolution obtained by HPLC.

  In order to quantify the differences between HPLC and the methods and equipment described here, equivalent separations were performed by HPLC and centrifugal liquid chromatography. As the stationary phase, particles each having a diameter of 5 microns were used. The length of the column used in HPLC is 150 mm. The length of the column used in the centrifugal liquid chromatography is 36 mm. In both methods, similar mobile phases and samples were used. The measured values of the experiment are shown in Table 2.

  The big difference between HPLC and centrifugal liquid chromatography is unexpected. The separation efficiency of centrifugal liquid chromatography is several orders of magnitude better than HPLC. Many explanations can be given that centrifugal liquid chromatography shows high separation efficiency. Apart from the individual theories about why typical results were obtained, here are some points I noticed.

  When centrifugal force is used to pass the fluid through the chromatographic enclosure, much less diffusion occurs as a result of the concentration gradient than when pressure is used as in HPLC. Less diffusion increases the separation efficiency, resulting in a higher number of theoretical plates in the examples described here as opposed to HPLC. The sedimentation coefficient associated with centrifugation depends on the concentration. For this reason, diffusion was reduced by centrifugation.

Applied chromatography is a fundamental tool for experimental chemists and those who apply chemistry to their respective disciplines. Chromatography is widely used and essential in the petroleum industry, food industry, pharmaceutical industry, medicine (eg, medical examination), and other industries. Looking more closely at the fields of application of the examples described here to chromatographic processing,
Medical and biomedical research Drug quality control Routine clinical analysis Drug screening Space-related research and development Geochemistry research and development Pharmaceutical research and development Criminal science Food and cosmetic chemical measurements Process control in the petroleum industry Environment Monitoring and pollution control Studies on chemistry and metabolism in biological systems Some specific examples of analyzes possible using the devices described here include human plasma proteins, nucleotides and derivatives, amino acids and derivatives , But not limited to, analysis of urinary metabolites, therapeutic drug monitoring, drug abuse monitoring, and wheat and other types of protein analysis.

  The two main purposes of chromatographic processing are distinguished by analysis and preparation by their size and purpose. Preparative chromatography is used to purify and collect a few samples in large quantities. Analytical chromatography is used to determine the composition and purity of many small samples. The following are examples of sample sizes, column lengths, and column inner diameter (ID) used for various preparations and analyses.

  The examples shown here can be used for both preparation and analysis. For large samples, such as industrial processes, the equipment and methods can be scaled up accordingly. Column size, such as column length and inner diameter, can be varied depending on the purpose as indicated above, and in various embodiments ranges from 10 to 400 cm and 0.1 to 2000 mm, respectively. The column length and inner diameter can be chosen to meet the particle requirements. For example, a capillary having an inner diameter of 0.01 mm or less can be used. Accordingly, the ranges set forth above are not intended to be limiting and are for illustrative purposes only. Rotating elements such as the rotor assembly described above can also be scaled up or down to accommodate specially sized columns for special applications.

Method FIG. 13 is a flowchart illustrating a method 700 for performing a chromatographic separation process. In step 702, a plurality of components are prepared: 1) a chromatographic enclosure containing a stationary phase, 2) a mobile phase fluid source, and 3) a sample source. At step 704, the chromatographic enclosure is coupled to a rotating element such as a rotor assembly.

  In step 706, the rotating element is rotated from rest to provide angular velocity to the chromatographic enclosure. During rotation, the mobile phase fluid supplied from the mobile phase fluid source by centrifugal force moves through the stationary phase. The mobile phase fluid source may be a component that is integral with the chromatographic enclosure or may be located separately from the chromatographic enclosure.

  At step 708, it is determined whether a mobile phase fluid flow has formed through the stationary phase during rotation of the chromatographic enclosure. Specifically, it may be determined whether a steady flow through the chromatographic enclosure has been formed. In one embodiment, measurements using a flow cell located near the end of the chromatographic enclosure may be used to determine whether a flow has been formed.

  In general, a detector can be used to determine whether the column has reached a steady condition. Examples of available detectors have been described above, but for example, the detectors described in the “Apparatus” section can be used. The signal from the detector, for example the signal generated by the mobile phase fluid exiting the column, is measured over a predetermined period. The chromatogram shown in FIG. 12 is an example of a signal obtained from the detector. The signal may contain noise. The stationarity may be defined as an average value of signals whose variation is less than a predetermined amount over a target period. A plurality of detectors may be associated with one column, and signals from the plurality of detectors may be used to determine whether a steady condition has been reached.

  Another factor that can be measured is the angular velocity of the rotor assembly. In some embodiments, the rotor assembly may be spun up from a resting state to a target angular velocity during a particular measurement, or the rotor assembly may be subjected to a first angular velocity during the first measurement. In the next measurement, the angular velocity may be increased or decreased to a new target value. A detector may be used to determine whether the average angular velocity of the rotor assembly is within a predetermined range over a predetermined period. For example, a motor connected to the rotor assembly may notify the rotation speed. In some embodiments, sample injection may not be initiated until it is determined that the average angular velocity is within a predetermined range over the period of interest. Also, in some embodiments, sample injection is initiated only when both the signal from one or more detectors related to the flow through the column and the signal related to angular velocity are within a predetermined range for a predetermined period of time. You may do it.

  As an example, in a series of measurements in a chromatographic system, a mobile phase fluid is first introduced into the column. The mobile phase fluid composition may generate a first signal output from the detector. When this signal changes with time within a predetermined range, for example, when the signal value changes relatively little, a sample may be injected to perform gradient elution. Gradient elution may be required to allow a given analyte to flow out of the column. Gradient elution changes the signal output from the detector. A change in signal due to elution can be detected as a recognizable profile such as a linear profile having a specific slope. A change in signal also occurs due to the outflow of the analyte from the column. Changes in the signal due to the analyte can be detected as recognizable profiles such as peaks and valleys. The signal change profile produced by the analyte is different from the signal change profile produced by gradient elution and it is also possible to distinguish the effects on individual signal changes.

  After gradient elution is complete, the mobile phase fluid returns to the original composition prior to gradient elution. In response, the signal from the detector returns to the initial value, i.e., the value before gradient elution is started, after a predetermined period of time. In some embodiments, 10 to 20 column volumes of fluid may flow through the column. If the signal returns to the initial value and the value is within a predetermined range for a predetermined period, the next sample can be injected into the column.

  At step 710, sample fluid is introduced from the sample source and passed through the stationary phase during rotation of the chromatographic enclosure. As the sample fluid moves through the stationary phase, one or more sample components can be chromatographically separated from the sample fluid. At step 712, during the rotation of the chromatographic enclosure, the separated components of the sample fluid are detected after passing through the stationary phase. For example, the separated component may be passed through a flow cell near the end of the chromatograph enclosure, and the presence of the separated component may be detected by measurement using the flow cell. The result of such measurement may be displayed as a chromatogram on an output device connected to the chromatograph system.

  The present invention has a number of advantages. Various aspects, embodiments or examples have one or more of the advantages described below. One advantage is that smaller stationary phase particles can be utilized by passing fluid through a chromatographic column using centrifugation than other types of chromatographic techniques such as HPLC. By using small particles, chromatographic separation efficiency can be improved as compared with other types of chromatographic techniques such as HPLC. Another advantage is that the rotor assembly described herein can perform multiple chromatographic processes simultaneously. Through “parallel processing” of a large number of chromatographic processes, the throughput time is shortened, and it is possible to perform an analysis that takes too much time and costs too much for other chromatographic processes such as HPLC. Many features and advantages of the present invention are apparent from the specification, and all such features and advantages of the invention are included within the scope of the invention as set forth in the claims. It is. Further, as is obvious to those skilled in the art, various modifications and changes are possible, and the present invention is not limited to the specific configurations and operations shown and described. Accordingly, all appropriate modifications and changes, and equivalents thereof, are included within the scope of the present invention.

Claims (159)

A centrifugal column chromatograph system,
A rotor configured to rotate about an axis;
A plurality of chromatographic column enclosures held by the rotor, each chromatographic column enclosure containing a corresponding chromatographic stationary phase and contained within the chromatographic column enclosure; Configured to facilitate the movement of fluid through the chromatographic stationary phase, and through the chromatographic stationary phase in the axial direction of the chromatographic column enclosure by centrifugal force generated by rotation of the rotor. Move multiple chromatographic column enclosures,
A sample introduction mechanism fluidly coupled to at least a selected one of the plurality of chromatographic column enclosures, wherein the selected chromatographic column enclosure is coupled to the selected chromatographic column enclosure while the rotor is rotating; A sample introduction mechanism for introducing a sample fluid;
An eluent reservoir held in the rotor and fluidly coupled to the plurality of chromatographic column enclosures, the eluent reservoir containing fluid eluted from the plurality of chromatographic column enclosures. Centrifugal column chromatograph system. A centrifugal chromatograph system,
A rotor configured to rotate about an axis;
A plurality of chromatographic enclosures held by the rotor, each chromatographic enclosure containing a corresponding chromatographic stationary phase and the chromatographic stationary contained within the chromatographic enclosure; A plurality of chromatographic enclosures configured to facilitate movement of fluid through the phase and moving the fluid through the chromatographic stationary phase by centrifugal force generated by rotation of the rotor;
An eluent reservoir held in the rotor and fluidly connected to the plurality of chromatographic enclosures, the eluent reservoir containing fluid eluted from the plurality of chromatographic enclosures, and a centrifugal chromatograph ·system. A centrifugal chromatograph system,
A rotor configured to rotate about an axis;
At least one chromatographic enclosure held by the rotor;
A mixing chamber that is held in the rotor and mixes a mobile phase fluid with a sample on the rotor to generate a mixed fluid, and the sample and the mobile phase are utilized by rotation of components held in the rotor. A centrifugal chromatographic system comprising: a mixing chamber that facilitates mixing with a fluid and disposed upstream of the chromatographic enclosure. The centrifugal chromatograph system according to claim 3, further comprising:
A mobile phase fluid reservoir fluidly coupled to the mixing chamber;
A sample introduction mechanism fluidly connected to the mixing chamber,
A centrifugal chromatograph system, wherein the mobile phase fluid reservoir and the sample introduction mechanism are disposed either on the rotor or outside the rotor. A centrifugal chromatograph system,
A chromatographic enclosure;
A rotor for holding the chromatographic enclosure and rotating the chromatographic enclosure;
A sample introduction mechanism fluidly coupled to the chromatographic enclosure, the sample introduction mechanism configured to introduce a sample fluid into the chromatographic enclosure while the rotor is rotating, receiving a sample introduction signal, and the sample A centrifugal chromatograph system comprising: a sample introduction mechanism that starts introduction of the sample fluid in response to reception of an introduction signal. A chromatographic system,
A chromatographic enclosure;
A rotor for holding the chromatographic enclosure and rotating the chromatographic enclosure;
A sample introduction mechanism fluidly coupled to the chromatographic enclosure, wherein the sample introduction mechanism introduces sample fluid into the chromatographic enclosure while the rotor is rotating;
And a controller connected to the sample introduction mechanism and automatically starting the introduction of the sample fluid. The centrifugal chromatograph system according to claim 6,
The centrifugal chromatograph system, wherein the controller initiates introduction of the sample fluid based at least in part on a sensing condition associated with the centrifugal chromatograph system. A chromatographic system,
A chromatographic enclosure containing a corresponding chromatographic stationary phase, the chromatographic enclosure configured to facilitate movement of fluid through the chromatographic stationary phase contained within the chromatographic enclosure. An enclosure,
A rotor for holding the chromatographic enclosure, wherein the chromatographic enclosure is rotated at a predetermined angular velocity so that fluid is moved in the chromatographic enclosure including the chromatographic stationary phase by centrifugal force. A configured rotor;
A fluid enclosure having a first portion stationary upon rotation of the rotor and a second portion retained on the rotor and rotating with the rotor and fluidly coupled to the chromatographic enclosure A chromatographic system comprising: a fluid enclosure; A centrifugal chromatograph system according to claim 8,
The centrifugal chromatographic system, wherein the fluid enclosure is a mobile phase fluid reservoir configured to hold a mobile phase fluid for supply to the chromatographic enclosure on the rotor. A centrifugal chromatograph system according to claim 8,
The centrifugal chromatograph system, wherein the fluid enclosure is an eluent reservoir configured to contain an eluent that has passed through the chromatographic enclosure on the rotor. A centrifugal chromatograph system,
A rotor holding at least one chromatographic enclosure, the rotor being configured to rotate the chromatographic enclosure to move the liquid within the at least one chromatographic enclosure by centrifugal force; ,
A fluid supply mechanism fluidly connected to the chromatographic enclosure for facilitating the supply of mobile phase fluid to the chromatographic enclosure, wherein the first portion is stationary when the rotor is rotated, and on the rotor And a second portion that rotates together with the rotor, and is configured to move fluid from the first portion of the fluid supply mechanism to the second portion when the rotor rotates. A centrifugal chromatograph system comprising: a supply mechanism; A centrifugal chromatograph system,
A rotor configured to rotate about an axis;
A chromatographic enclosure, retained on the rotor, containing a corresponding chromatographic stationary phase and facilitates fluid movement through the chromatographic stationary phase contained within the chromatographic enclosure. A chromatographic enclosure configured to move the fluid within the chromatographic stationary phase by centrifugal force generated by rotation of the rotor;
A flow cell fluidly coupled to the chromatographic enclosure and provided with a flow window, the flow path passing through the chromatographic enclosure and the flow cell, wherein the predetermined channel within the chromatographic stationary phase of the chromatographic enclosure A centrifugal chromatographic system, wherein the inner cross-sectional area of the flow path starting from position and passing through the flow window is substantially constant. A flow cell suitable for use in a chromatographic system,
A flow cell inlet, a flow cell outlet, and a flow path extending between the flow cell inlet and the flow cell outlet;
A window formed along the flow path and enabling optical detection of a substance passing through the flow cell;
The sectional dimension of the entrance is different from the sectional dimension of the window,
The flow cell, wherein a cross-sectional area of the inlet, a cross-sectional area of the window, and a cross-sectional area of a region between the inlet and the window are substantially constant along the flow path. A flow cell according to claim 13,
The flow cell, wherein a cross-sectional area of the flow path is substantially constant throughout the flow cell from the flow cell inlet to the flow cell outlet. A chromatographic system,
A chromatographic column containing a chromatographic stationary phase;
A flow cell according to claim 13,
The flow cell is disposed downstream of the chromatographic column;
A chromatographic system, wherein the cross-sectional dimension and cross-sectional area of the chromatographic column are substantially the same as the cross-sectional dimension and cross-sectional area of the flow cell inlet. A chromatographic system according to claim 15,
The chromatographic system, wherein the chromatographic column is held on a rotor, and the rotor applies a centrifugal force to move a mobile phase through the stationary phase. A centrifugal column chromatograph system,
A rotor configured to rotate about an axis;
A plurality of chromatographic column enclosures retained on the rotor, each chromatographic column enclosure containing a corresponding chromatographic stationary phase and within the chromatographic column enclosure A plurality of chromatographs configured to facilitate movement of fluid through the contained chromatographic stationary phase, wherein the fluid is moved within the chromatographic stationary phase by centrifugal force generated by rotation of the rotor; A column enclosure;
A reservoir comprising a container with a cover for containing at least a mobile phase fluid, the reservoir being disposed in proximity to a rotation axis, retained on the rotor, and fluidly connected to each of the plurality of chromatographic column enclosures; A centrifugal column chromatograph system. A chromatographic system,
A rotor configured to rotate about an axis;
A chromatographic enclosure held by the rotor, wherein the first end is located at a first distance from the axis and the second end is located at a second distance greater than the first distance from the axis. A chromatographic enclosure comprising:
A flow cell fluidly coupled to a chromatographic enclosure disposed proximate to the second end held on the rotor;
A load bearing mechanism connected to the chromatographic enclosure and the rotor, which reduces the mechanical load transmitted by the chromatographic enclosure into the flow cell during rotation and reduces the mechanical load And a load bearing mechanism configured to prevent deformation of the flow cell window in the flow cell. A chromatographic system,
A rotor configured to rotate about an axis;
A chromatographic enclosure held by the rotor;
A containment structure surrounding the rotor;
A plurality of rotor support structures supported by the containment structure, each rotor support structure comprising a gas bearing, wherein the gas bearings cooperate to stabilize the rotor as the rotor rotates. ·system. A chromatographic system according to claim 19, comprising:
At least a portion of the rotor support structure functions as an instrument attachment, and each instrument attachment is discharged from the chromatographic enclosure and passes through a flow cell disposed downstream of the chromatographic enclosure. A chromatographic system configured to support a corresponding instrument suitable for detecting a characteristic. A centrifugal chromatograph system,
A rotor comprising a chromatographic enclosure held by the rotor;
A gas chromatographic system comprising: a gas bearing proximate to the rotor, the gas bearing configured to stabilize the rotor during rotation of the rotor. The centrifugal chromatograph system according to claim 21,
The centrifugal chromatograph system, wherein the gas bearing does not rotate with the rotor. The centrifugal chromatograph system according to claim 21, further comprising:
A centrifugal chromatograph system comprising a gas reservoir fluidly coupled to the gas bearing, the gas reservoir supplying gas released from the gas bearing. A chromatographic system,
A plurality of chromatographic enclosures retained on the rotor, each chromatographic enclosure containing a corresponding chromatographic stationary phase and contained within the chromatographic enclosure; A plurality of chromatographs configured to facilitate movement of fluid through the phases, wherein the fluid is moved through the chromatographic stationary phase toward an outer peripheral surface of the rotor by centrifugal force generated by rotation of the rotor;・ Enclosure,
A plurality of links arranged around the outer peripheral surface of the rotor, each link being connected to two other links to form a continuous chain ring around the outer peripheral surface of the rotor. One or more of the links are configured to 1) contain fluid moving toward the outer peripheral surface, and 2) change the direction of the fluid away from the outer peripheral surface and inwardly. A plurality of links comprising: a chromatographic system comprising: A chromatographic system according to any preceding claim,
The rotor is a chromatographic system that holds a plurality of chromatographic column enclosures. A chromatographic system according to any preceding claim,
Each chromatographic enclosure contains a corresponding chromatographic stationary phase and is configured to facilitate fluid movement through the chromatographic stationary phase contained within the chromatographic enclosure, wherein the rotor A chromatographic system in which the fluid is moved in the chromatographic stationary phase by centrifugal force generated by rotation of the. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a rotor drive system that provides angular velocity to the rotor. A chromatographic system according to any preceding claim,
The chromatographic system, wherein the chromatographic enclosure is integrally formed with the rotor. A chromatographic system according to any preceding claim,
Each chromatographic enclosure comprises a chromatographic system comprising at least one hollow inner portion containing said chromatographic stationary phase. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising at least one mobile phase fluid reservoir fluidly coupled to one or more of the chromatographic enclosures. A chromatographic system according to claim 30, comprising
The chromatographic system, wherein the at least one mobile phase fluid fluid reservoir is retained on the rotor. The chromatographic system according to any preceding claim further comprises:
At least one sample introduction mechanism fluidly coupled to one or more of the chromatographic enclosures, wherein the at least one sample is configured to introduce sample fluid into the chromatographic enclosure upon rotation of the rotor; A chromatographic system with an introduction mechanism. A chromatographic system according to claim 32, comprising:
The chromatograph system, wherein the sample introduction mechanism is held on the rotor. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a controller configured to automatically initiate introduction of sample fluid into the chromatographic enclosure. A chromatographic system according to claim 34, comprising:
The controller detects the state of the fluid held by the rotor and samples at any point during rotation of the rotor based at least in part on the detected state of the fluid held by the rotor. A chromatographic system that determines whether a fluid should be introduced. 36. A chromatographic system according to claim 35, comprising:
The chromatograph system, wherein the controller is held on the rotor. The chromatographic system according to any preceding claim further comprises:
A chromatograph comprising at least one eluent reservoir fluidly coupled to one or more of said chromatographic enclosures, wherein said eluent reservoir contains fluid eluted from said one or more chromatographic enclosures. Graph system. A chromatographic system according to claim 37, comprising:
A chromatographic system, wherein the eluent reservoir is held on the rotor. A chromatographic system according to claim 37 or 38,
The chromatographic system, wherein the eluent reservoir comprises two or more chambers. A chromatographic system according to any of claims 37, 38 or 39,
A chromatographic system, wherein the volume of the eluent reservoir is variable. The chromatographic system according to any of claims 37, 38, 39 or 40 further comprises:
A chromatographic system comprising a volume control mechanism connected to the eluent reservoir, the volume control mechanism controlling a volume of the eluent reservoir. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a sensor for detecting a liquid level in the eluent reservoir. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising at least one detection mechanism for measuring a physical property of fluid discharged from one or more of the chromatographic enclosures. The chromatographic system according to any preceding claim further comprises:
A plurality of detection mechanisms, each of which detects a physical property of the fluid discharged from the chromatographic enclosure;
A chromatographic system, wherein at least one of the detection mechanisms is held on the rotor. 45. A chromatographic system according to claim 44, comprising:
At least one of the detection mechanisms includes a laser-induced fluorescence (LIF) detector, a refractive index detector, a UV (Ultra-Violet) absorbance detector, and a visible light (Vis) visible absorbance detector. A chromatographic system selected from the group consisting of: a scanning UV-Vis detector, a radiochemical detector, an electrochemical detector, a mass spectrometry detector, a nuclear magnetic resonance detector, and a light scattering detector. A chromatographic system according to any of claims 44 or 45, comprising:
At least one of the detection mechanisms generates a signal related to a physical property of the fluid discharged from the chromatographic enclosure;
The chromatographic system further comprises a display mechanism for outputting information related to physical properties of the fluid. The chromatographic system according to any of claims 44, 45 or 46 further comprises:
Comprising a processor designed or configured to process data related to the detection mechanism;
A chromatographic system, wherein the processor is held on the rotor. The chromatographic system according to any preceding claim further comprises:
A flow cell held on said rotor and configured to contain fluid discharged from at least one chromatographic enclosure and having a flow window;
A flow path through the chromatographic enclosure and the flow cell, the inner cross-sectional area of the flow path starting from a predetermined position in the chromatographic stationary phase of the chromatographic enclosure and passing through the flow window is substantially A chromatographic system that is constant. The chromatographic system according to any preceding claim further comprises:
A containment structure surrounding the rotor;
A chromatographic system comprising: a plurality of instrument mounting portions connected to the storage structure, the plurality of instrument mounting portions not rotating together with the rotor. 50. A chromatographic system according to claim 49, comprising:
Each said instrument mounting part is a chromatograph system attached to the said storage container by a retractable arm. A chromatographic system according to any of claims 49 or 50,
Each of the instrument mountings further comprises a gas bearing, wherein the gas bearing stabilizes the rotor as the rotor rotates in proximity to the instrument mounting. The chromatographic system according to any of claims 49, 50 or 51 further comprises:
A flow cell held in the rotor and configured to contain fluid that has passed through at least a portion of at least one chromatographic enclosure and having a flow window;
Each of the instrument mounting portions further includes a light source and a light detection mechanism, and the light source and the light detection mechanism are arranged on opposite sides of the rotor, and the light source emits light toward the light detection mechanism. Configured to communicate,
A chromatographic system in which when the rotor rotates, the flow cell periodically passes between the light source and the light detection mechanism so that the light detection mechanism detects characteristics of the fluid passing through the flow cell. A chromatographic system according to claim 52, comprising:
The photodetection mechanism is a chromatographic system, which is one of a photomultiplier tube, a photodiode, a photodiode array, and a CCD (charge coupled device). A chromatographic system according to any of claims 52 or 53, wherein
The rotor holds various chromatographic column enclosures, each chromatographic column enclosure has a corresponding flow cell, and the flow cell corresponding to each chromatographic column is separated from the rotation axis of the rotor. A chromatographic system in which the light detection mechanism detects characteristics of the fluid passing through each flow cell once per rotation of the roller by being disposed at substantially the same radial distance. The chromatographic system according to any preceding claim further comprises:
A reaction chamber fluidly connected to the chromatographic enclosure and one or more reagent reservoirs for receiving fluid discharged from the chromatographic enclosure and reacting the one or more reagents with the fluid. A chromatographic system comprising: A chromatographic system according to claim 55, wherein
A chromatographic system, wherein the reaction chamber is held on the rotor. The chromatographic system according to any of claims 55 or 56 further comprises:
A chromatographic system comprising an ultrasonic vibration generator connected to the reaction chamber, the ultrasonic vibration generator for promoting a chemical reaction in the reaction chamber. The chromatographic system according to any of claims 55, 56 or 57 further comprises:
Comprising an energy source connected to the reaction chamber;
A chromatographic system that promotes a chemical reaction in the reaction chamber by applying energy from the energy source to the reaction chamber. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a power source connected to the rotor for supplying power to one or more devices held on the rotor. 60. A chromatographic system according to claim 59, comprising:
The chromatograph system, wherein the power source includes a battery. The chromatographic system according to any preceding claim further comprises:
Equipped with a generator,
The generator includes a first portion of the generator held on the rotor and a second portion of the generator disposed outside the rotor, the first portion and the second portion. A chromatographic system in which the interaction between and generates an available current on the rotor. The chromatographic system according to any preceding claim further comprises:
With an electrical interface,
The electrical interface comprises a first portion of the electrical interface held on the rotor and a second portion of the electrical interface disposed outside the rotor, and during rotation of the rotor, A chromatographic system, wherein the first part and the second part are in contact and power is transferred between the first part and the second part. The chromatographic system according to any preceding claim further comprises:
A communication interface held on the rotor for establishing communication between a first device held on the rotor and a second device arranged outside the rotor; Equipped with a chromatographic system. A chromatographic system according to claim 63, comprising:
The chromatograph system, wherein the communication interface is a wireless communication interface. A chromatographic system according to any preceding claim,
The chromatographic stationary phase is composed of a nonpolar substance that can be used for reverse phase chromatographic processing,
A chromatographic system wherein the mobile phase fluid is capable of reverse phase chromatographic processing. A chromatographic system according to any preceding claim,
The chromatographic system, wherein the chromatographic enclosure comprises a single conduit through which fluid flows along its length. A chromatographic system according to any of claims 1 to 65, wherein
The chromatographic enclosure comprises a plurality of conduits through which fluid flows along its length, each of the plurality of conduits providing a separate flow path through the chromatographic enclosure. system. A chromatographic system according to any of claims 66 or 67, wherein
A chromatographic system, wherein the internal area of each conduit is constant along the length. A chromatographic system according to any preceding claim,
The chromatographic system, wherein the chromatographic enclosure comprises a cylindrical column having a length and an inner diameter. A chromatographic system according to claim 69, comprising:
The chromatographic system wherein the column has a length in the range of 10 to 400 cm and an inner diameter in the range of 0.02 to 2000 mm. A chromatographic system according to any preceding claim,
A chromatographic system, wherein the maximum working pressure of the fluid in the flow path retained on the rotor is less than 100 PSI. A chromatographic system according to any preceding claim,
The chromatographic system, wherein the chromatographic stationary phase has a particle size of less than about 1 micron. A chromatographic system according to any preceding claim,
The chromatographic system, wherein the chromatographic stationary phase has a particle size in the range of about 10 angstroms to 1 micron. The chromatographic system according to any one of the preceding claims,
A chromatographic system comprising a sensor for detecting leakage of fluid from a flow path held on the rotor. The chromatographic system according to any preceding claim further comprises:
A reservoir comprising a covered container for containing at least a mobile phase fluid, the reservoir comprising a reservoir disposed proximate to the axis of rotation, retained on the rotor, and fluidly connected to each of a plurality of chromatographic enclosures. Graph system. A chromatographic system according to any preceding claim,
A chromatographic system in which the same chromatographic stationary phase material is used in two or more of the plurality of chromatographic enclosures. A chromatographic system according to any of claims 1 to 75,
A chromatographic system in which different chromatographic stationary phase materials are used in two or more of the plurality of chromatographic enclosures. A chromatographic system according to any preceding claim,
A chromatographic system, wherein two or more of the plurality of chromatographic enclosures are fluidly coupled to a shared reservoir and receive mobile phase fluid from the shared reservoir. A chromatographic system according to any preceding claim,
Two or more of the plurality of chromatographic enclosures are fluidly coupled to a shared sample introduction mechanism, and the shared sample introduction mechanism is configured to introduce a common sample fluid to each of the two or more chromatographic enclosures. The chromatographic system. The chromatographic system according to any preceding claim further comprises:
1) a plurality of chromatographic enclosures, and 2) a mixing chamber,
A chromatographic system, wherein each of the chromatographic enclosures is fluidly coupled to the mixing chamber, the mixing chamber being held on the rotor. A chromatographic system according to claim 80, comprising:
The mixing chamber is fluidly coupled to one or more mobile phase reservoirs, and each of the chromatographic enclosures is stored in the one or more mobile phase reservoirs after the mobile phase fluid is mixed in the mixing chamber. A chromatographic system for receiving said mobile phase fluid being applied. A chromatographic system according to any of claims 80 or 81, wherein
The chromatographic system, wherein the mixing chamber comprises a plurality of mixing pins that mix the fluid entering the mixing chamber. A chromatographic system according to any of claims 80 to 82, wherein
The chromatographic system, wherein the mixing chamber is configured to mix the sample fluid received from the sample introduction mechanism and the mobile phase fluid. A chromatographic system according to any preceding claim,
The rotor includes a disk portion extending substantially perpendicular to a rotation center axis of the rotor, and a plurality of chromatographic enclosures are held on the disk portion, and a fluid is rotated in each chromatographic enclosure. A chromatographic system that moves away from the central axis. The chromatographic system according to any preceding claim further comprises:
A coupling mechanism provided between one or more of the chromatographic enclosures and the rotor, wherein an arrangement of the one or more chromatographic enclosures with respect to the rotor is different from that of the one or more chromatographic enclosures; A chromatographic system comprising a coupling mechanism for changing from a first position where the rotor is at rest to a second position where the one or more chromatographic enclosures and the rotor are rotating. A chromatographic system according to any preceding claim,
During the rotation of the rotor, the fluid in one or more flow paths held on the rotor is pressurized and the one or more flows to prevent leakage of the fluid from the one or more flow paths. A chromatographic system that seals the road. The chromatographic system according to any preceding claim further comprises:
A fluid enclosure having 1) a fixed first portion remote from the rotor; and 2) a second portion retained on the rotor;
During rotation of the second portion, the fluid enclosure is configured to receive a mobile phase fluid or sample fluid from outside the rotor and supply the received fluid to the chromatographic enclosure. . The chromatographic system of claim 87 further comprises:
A chromatographic system comprising a fluid line connected to the fixed first portion of the fluid enclosure, the fluid line configured to supply the fluid to the fluid enclosure. The chromatographic system of claim 87 further comprises:
A plurality of fluid conduits connected to the fixed first portion of the fluid enclosure, each comprising a plurality of fluid conduits configured to supply fluid to the fluid enclosure. system. A chromatographic system according to any of claims 87 to 89, wherein
The chromatographic system, wherein the second portion of the fluid enclosure comprises one or more structures configured to mix fluid within the fluid enclosure during rotation of the second portion. A chromatographic system according to any one of claims 87 to 90, comprising:
The chromatographic system, wherein the first portion of the fluid enclosure further comprises a gas bearing. A chromatographic system according to claim 91, wherein
The chromatographic system, wherein the second portion of the fluid enclosure further comprises a surface on which the gas bearing rests upon rotation of the second portion. A chromatographic system according to any one of claims 87 to 92, comprising:
A chromatographic system wherein forces that stabilize the rotor are transmitted through the first portion of the fluid enclosure to the second portion of the fluid enclosure during rotation of the rotor. The chromatographic system according to any preceding claim further comprises:
An effluent fluid enclosure having a first portion retained on the rotor, and 2) a fixed second portion remote from the rotor,
During the rotation of the first portion, the effluent fluid enclosure is configured to receive fluid from the rotor and supply the received fluid out of the rotor. The chromatographic system according to any preceding claim further comprises:
A sample introduction mechanism configured to discharge sample fluid;
A control device connected to the sample introduction mechanism,
The control device 1) receives a signal from the detection mechanism, 2) determines that the change in the signal is within a predetermined range over a predetermined period, and 3) the signal is within the predetermined range over the predetermined period. If so, a chromatographic system that is designed or configured to instruct the sample introduction mechanism to release the sample fluid. 96. A chromatographic system according to claim 95, comprising:
The chromatographic system, wherein the signal received from the detection mechanism is related to characteristics of the fluid that has passed through the chromatographic enclosure. The chromatographic system of claim 96 further comprises:
A chromatographic system comprising a second detection mechanism configured to generate a signal related to the angular velocity of the rotor. A chromatographic system according to claim 97,
The control device further includes 1) a signal related to the angular velocity from the second detection mechanism, and 3) both the signal received from the detection mechanism and the signal received from the first detection mechanism. A chromatographic system that is designed or configured to command the sample introduction mechanism to release the sample fluid when within the predetermined range for the predetermined period of time. The chromatographic system according to any preceding claim further comprises:
A sample introduction mechanism configured to discharge sample fluid;
A control device connected to the sample introduction mechanism,
The controller 1) determines whether the interior of the chromatograph enclosure has reached a steady condition, and 2) based on the determination of whether the steady condition has been reached, based at least in part on the sample fluid. A chromatographic system designed or configured to issue instructions to the sample introduction mechanism. The chromatographic system of claim 99 further comprises:
A first detection mechanism communicably connected to the control device, the first detection mechanism generating a signal related to an operating characteristic of the chromatographic system that changes over time;
A chromatographic system, wherein the determination of whether or not a steady-state condition has been reached by the controller is made based at least in part on a signal received from the first detection mechanism. A chromatographic system according to claim 100, comprising:
The chromatographic system, wherein the operating characteristic is related to a physical characteristic of the fluid that has passed through the chromatographic enclosure. A chromatographic system according to claim 100, comprising:
The chromatographic system, wherein the operating characteristic is related to the angular velocity of the rotor. The chromatographic system according to any preceding claim further comprises:
A closed flow path containing pressurized fluid on the rotor, 1) a first portion for flowing the fluid from an inlet of the flow path and moving the fluid away from the axis; 2) a second portion fluidly connected to the first portion, moving the fluid in a direction toward the axis, and flowing out from the outlet of the flow path, wherein the outlet is a first radial distance from the shaft; And when the liquid surface in the first portion is located at a second radial distance that is smaller than the first radial distance during rotation of the rotor, the fluid is A chromatograph comprising a closed flow path comprising a second portion continuously discharged from the outlet; and 3) the chromatographic enclosure disposed between the inlet and the outlet of the flow path. Graph system. A chromatographic system according to claim 103, wherein
The flow path includes a plurality of portions,
Each of the plurality of portions causes the fluid to flow in from separate inlets and move the fluid away from the axis;
The chromatographic system, wherein the second portion is fluidly connected to each of the plurality of portions. The chromatographic system according to any preceding claim further comprises:
A flow cell in fluid communication with the chromatographic enclosure;
A flow path connecting between the flow cell and the chromatographic enclosure, starting from a predetermined position within the chromatographic stationary phase of the chromatographic enclosure, through the chromatographic enclosure and into the flow cell A chromatographic system in which the internal area of the flow path that enters, passes through the flow window of the flow cell and ends at a predetermined position in the flow cell is constant. The chromatographic system according to any preceding claim further comprises:
A load-bearing mechanism connected to the chromatographic enclosure and the rotor, by reducing a mechanical load transmitted by the chromatographic enclosure into the flow cell during rotation, and reducing the mechanical load A chromatographic system comprising a load bearing mechanism configured to prevent deformation of a flow cell window in the flow cell due to a load transmitted from the chromatographic enclosure to the flow cell. A chromatographic system according to any preceding claim,
A chromatographic system, wherein a portion of the fluid conduit retained on the rotor is flexible. A chromatographic system according to any preceding claim,
A plurality of chromatographic enclosures and a reservoir fluidly connected to the plurality of chromatographic enclosures share a common housing. A chromatographic system according to any preceding claim,
The chromatograph system, wherein the rotor includes a flow path branched into a plurality of flow paths, and each of the plurality of flow paths supplies a fluid to a separate chromatographic enclosure. A chromatographic system according to any preceding claim,
The rotor includes a plurality of flow paths that merge into one flow path on the downstream side of the chromatographic enclosure. A chromatographic system according to any preceding claim,
A chromatographic system, wherein the orientation of one or more of the chromatographic enclosures is fixed relative to the axis of rotation. A chromatographic system according to any preceding claim,
A chromatographic system, wherein one or more of the chromatographic enclosures extend radially and perpendicular to the axis of rotation. The chromatographic system according to any preceding claim further comprises:
One or more valves for controlling the fluid on the rotor;
A chromatographic system comprising one or more electronically controllable actuation mechanisms that change the state of the one or more valves. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising one or more pumps that move fluid from one position to another within the chromatographic system. A chromatographic system according to claim 114, wherein
The chromatographic system, wherein the one or more pumps are held on the rotor. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a rotation management system for controlling the angular velocity of the rotor. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising an indexing system that identifies one or more of the chromatographic enclosures while the rotor is rotating. The chromatographic system according to any preceding claim further comprises:
A gradient forming unit providing a mobile phase fluid containing two or more different fluids, the ratio of each of the two or more different fluids supplied to the one or more chromatographic enclosures varying over time A chromatographic system comprising a gradient former configured to cause The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising at least one environmental control mechanism configured to control at least a temperature of at least one chromatographic enclosure, a fluid in the chromatographic enclosure, or a combination thereof. 120. A chromatographic system according to claim 119, comprising:
A chromatographic system, wherein the environmental control mechanism is held on the rotor. A chromatographic system according to any of claims 119 or 120, wherein
The chromatographic system, wherein the environmental control mechanism is configured to maintain the temperature at a target level. The chromatographic system according to any preceding claim further comprises:
A sealed enclosure surrounding the rotor;
A chromatographic system that maintains a sub-atmospheric pressure within the sealed enclosure while the rotor is rotating. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a gas bearing configured to transmit a load to the rotor to stabilize rotation of the rotor as the rotor rotates. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a flow management system that controls at least one or more valves retained on the rotor. The chromatographic system according to any preceding claim further comprises:
A data management system connected to one or more detection mechanisms, 1) receiving data from the one or more detection mechanisms, 2) generating a chromatogram, and 3) from the one or more detection mechanisms A chromatographic system comprising a data management system configured to store the received data and the chromatogram. A chromatographic system according to any preceding claim,
The chromatographic enclosure comprises: 1) an inlet through which the fluid enters the chromatographic enclosure; and 2) an outlet through which the fluid exits the chromatographic enclosure;
A chromatographic system, wherein the fluid pressure at the inlet is less than the fluid pressure at the outlet. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a flow path fluidly connected to a chromatographic enclosure formed integrally with the rotor. The chromatographic system according to any preceding claim further comprises:
A chromatographic system comprising a temperature control mechanism configured to control the temperature of at least one chromatographic enclosure. The chromatographic system according to any preceding claim further comprises:
It has a mobile phase fluid that can handle chromatographic processing.
The chromatographic system, wherein the chromatographic processing includes one or more of reverse phase partitioning, adsorption, anion exchange, cation exchange, size exclusion, gel filtration, affinity interactions, or combinations thereof. A method for operating a centrifugal chromatograph system, comprising:
Rotate the rotor that holds the chromatographic column,
Forming a flow of mobile phase fluid through the chromatographic column and passing the mobile phase through the chromatographic column by centrifugal force;
Introducing the sample fluid into the chromatographic column by introducing the sample fluid into the mobile phase fluid stream, and introducing the sample fluid as the rotor rotates. The method of operating the centrifugal chromatograph system of claim 130 further comprises:
Starting the release of the sample fluid. The method according to any of claims 130 or 131 further comprises:
Determine whether the chromatographic column has reached a steady flow condition of the mobile phase fluid,
A method of releasing the sample fluid after it is determined that the inside of the chromatographic column has reached a steady flow condition of a mobile phase fluid. A method for operating a centrifugal chromatograph system, comprising:
Rotate the rotor that holds the chromatographic column containing the chromatographic stationary phase,
During the rotation of the rotor, mobile phase fluid is supplied to the chromatographic column from a stationary mobile phase fluid reservoir,
Passing the mobile phase through the chromatographic column containing the stationary phase by centrifugal force. A method for operating a centrifugal chromatograph system, comprising:
Rotate the rotor that holds the chromatographic column containing the chromatographic stationary phase,
During rotation of the rotor, sample fluid is supplied to the chromatographic column from a stationary sample fluid reservoir,
Passing the sample fluid through the chromatographic column containing the chromatographic stationary phase by centrifugal force. A method for operating a centrifugal chromatograph system, comprising:
Rotating a rotor holding a plurality of chromatographic columns each containing a chromatographic stationary phase;
A mobile phase fluid is passed through the chromatographic column;
A method of collecting fluid that has passed through the plurality of chromatographic columns in a common reservoir held by the rotor. A method for operating a centrifugal chromatograph system, comprising:
Rotating a rotor holding a plurality of chromatographic columns each containing a chromatographic stationary phase;
During rotation of the rotor, the sample and mobile phase fluid are mixed in a mixing chamber held by the rotor to form a mixed sample;
A method wherein the plurality of chromatographic columns process the same sample in parallel by passing different portions of the mixed sample from the mixing chamber through each of the plurality of chromatographic columns. A method for operating a centrifugal chromatograph system, comprising:
1) rotate a rotor holding a plurality of chromatographic columns each containing a chromatographic stationary phase, and 2) a sealed fluid reservoir,
Supplying a fluid from the sealed fluid reservoir to each of the plurality of chromatographic columns. A method for operating a centrifugal chromatograph system, comprising:
1) rotate a rotor holding a plurality of chromatographic columns each containing a chromatographic stationary phase, and 2) a fraction reservoir,
A mobile phase fluid is passed through the chromatographic column;
Collecting the fraction of fluid that has passed through at least one of the chromatographic columns in the fraction reservoir; A method for operating a centrifugal chromatograph system, comprising:
Prepare multiple non-rotating detectors,
1) a plurality of chromatographic columns each containing a chromatographic stationary phase; and 2) a plurality of flow cells, each of the flow cells being arranged close to the end of each column, Rotating a rotor holding a plurality of flow cells receiving fluid discharged from one of the graph columns;
A method of detecting a physical property of the fluid in each of the plurality of flow cells once for each rotation of the rotor for each of the plurality of non-rotation detectors. The method of operating the centrifugal chromatograph system of claim 139 further comprises:
Activating only a portion of the non-rotating detector. A method for operating a centrifugal chromatograph system, comprising:
Rotating a rotor holding a plurality of chromatographic columns each containing a chromatographic stationary phase;
A method wherein at least two different samples are passed through at least one chromatographic column while each of the samples is introduced at a different time while continuously rotating the rotor. A method of operating the centrifugal chromatograph system of claim 141, comprising:
A method wherein between 10 and 20 column volumes of fluid are passed through the one chromatographic column between the introduction of the first sample and the introduction of the second sample. A method for operating a centrifugal chromatograph system, comprising:
Rotating a rotor holding a plurality of chromatographic columns each containing a chromatographic stationary phase;
Stabilizing the rotor with one or more gas bearings. The method according to any of the claims further comprises:
A method of causing gradient elution in the chromatographic column by changing the composition of the mobile phase fluid over time. The method according to any of the claims further comprises:
A method of determining whether a steady condition has been reached based on a change over time in physical properties of fluid discharged from the chromatographic column. 145. The method of claim 145, comprising:
A method of measuring the physical characteristics using a flow cell proximate to an outlet of the chromatographic column. The method according to any of the claims further comprises:
A method for determining whether a steady condition has been reached based on a change in angular velocity of the rotor with time. A method according to any of the preceding claims,
A method wherein a sample reservoir for storing sample fluid is retained on the rotor. A method according to any of the preceding claims,
A method wherein a mobile phase fluid reservoir for storing mobile phase fluid is retained on the rotor. The method according to any of the claims further comprises:
Mixing the mobile phase fluid sample fluid in a closed mixing chamber held on the rotor. The method according to any of the claims further comprises:
Supplying a mobile phase fluid from a stationary mobile phase reservoir to the rotor during rotation of the rotor. The method according to any of the claims further comprises:
Supplying a sample fluid from a stationary sample fluid reservoir to the rotor during rotation of the rotor. The method according to any of the claims further comprises:
Storing the fluid eluted from the chromatographic enclosure in a containment structure during rotation of the rotor. 153. The method of claim 153, comprising:
The method wherein the containment structure is held on the rotor. 153. A method according to any of claims 153 or 154, comprising:
The method wherein the fluid eluted from the chromatographic column comprises a fraction separated from the sample fluid. The method according to any of the claims further comprises:
Receive data from one or more detection mechanisms,
2) Generate a chromatogram,
3) A method of storing the data received from the one or more detection mechanisms and the chromatogram. The method according to any of the claims further comprises:
Changing the operating state of one or more valves retained on the rotor to affect changes in the flow path on the rotor. The method according to any of the claims further comprises:
Transmitting a command to a sample introduction mechanism and releasing the sample fluid that moves to the chromatographic column in response to receiving the command. The method according to any of the claims further comprises:
A method for controlling the temperature of one of the chromatographic enclosures.