WO2004085020A2

WO2004085020A2 - Apparatus for countercurrent chromatography

Info

Publication number: WO2004085020A2
Application number: PCT/US2004/007934
Authority: WO
Inventors: Yoichiro Ito
Original assignee: The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services
Priority date: 2003-03-21
Filing date: 2004-03-16
Publication date: 2004-10-07
Also published as: WO2004085020A3

Abstract

A plate apparatus for use in countercurrent chromatography comprises a disk shaped tube support having a spiral groove formed in an upper surface (63), at least one slot extending at least partially through the disk connecting an inner opening to an inner groove portion and an outer edge to an outer groove portion, and at least one layer of fluid flow tubing (80) accommodated substantially within the spiral groove and the slot. The tube support further comprises a radial return path from the outer portion of the spiral groove to the inner portion of the spiral groove. In as additional embodiment, the tube support comprises a plurality of interleaved spiral grooves wherein a single or plurality of layers of fluid flow tubing (80) are positioned in the interleaved spiral grooves.

Description

APPARATUS FOR COUNTERCURRENT CHROMATOGRAPHY

Background of the Invention Field of the Invention

[0001] The invention relates to countercurrent chromatography systems, and more particularly to an improved instrument design for use in countercurrent chromatography.

Description of the Related Art

[0002] Chromatography is a separation process that is achieved by distributing the substances to be separated between a mobile phase and a stationary phase. Those substances distributed preferentially in the moving phase pass through the chromatographic system faster than those that are distributed preferentially in the stationary phase. As a consequence, the substances are eluted from the column in inverse order of their distribution coefficients with respect to the stationary phase.

[0003] Chromatography is widely used for the separation, identification, and determination of the chemical components in a complex mixture. Chromatographic separation can be utilized to separate gases, volatile substances, nonvolatile material, polymeric material, and a wide variety of organic and biological substances.

[0004] The performance of countercurrent chromatography systems depends largely on the amount of stationary phase retained in the column, which determines both the resolving power of the solute peaks and the sample loading capacity. Numerous countercurrent chromatography systems have been developed to optimize the retention of the stationary phase of a sample in the column. The maximum attainable retention level tends to fall sharply with the application of higher flow rates of the mobile phase, resulting in loss of peak resolution. Consequently, the applicable flow rate has become one of the major limiting factors in countercurrent chromatography.

[0005] Some countercurrent chromatography systems utilize a complex hydrodynamic motion in two solvent phases within a column comprising a rotating coiled tube. If, for example, a horizontally mounted coil is filled with water and is rotated around its own axis, any object, either heavier or lighter than the water present in the column will tend to move toward one end of the coil. This end is then called the "head" and the other end, the "tail" of the coil. When the coil is filled with two immiscible solvent phases, the rotation establishes a hydrodynamic equilibrium between the two solvent phases, where the two phases are distributed in each turn at a given volume ratio (equilibrium volume ratio) and any excess of either phase remains at the respective tail of the coil for each solvent.

[0006] When the coil is filled with one of the solvents as a stationary phase and the other solvent is eluted from the coil from its head end, the hydrodynamic equilibrium tends to maintain the original equilibrium volume ratio of the two phases in the coil and thereby a certain volume of the stationary phase is permanently retained in the coil while the two phases are undergoing vigorous agitation with rotation of the coil. As a result, the sample solutes present in one phase and introduced locally at the inlet of the coil are subjected to an efficient partition process between the two phases and are chromatographically separated according to their partition coefficients.

[0007] In some cases, countercurrent chromatography utilizes a multi-layer coil as a separation column to produce a high efficiency separation with relatively favorable retention of the stationary phase in many solvent systems. Thus, countercurrent chromatography has been employed to achieve efficient separation of compounds in a sample solution under relatively high flow rates.

[0008] Previous column designs have relied on the use of a helical coil of tubing. U.S. Patent No. 4,430,216, hereby incorporated by reference in its entirety, describes a preparative countercurrent chromatography utilizing a multiple layer coiled column. The coiled column design includes a length of plastic tubing wound around a coil holder to form multiple layers of the coil. Although this system works reasonably well for some solvents, these systems often fail to retain a satisfactory amount of the stationary phase for highly viscous, low interfacial solvent systems such as polymer phase systems, which are useful for the separation of macromolecules and particulates. hi addition, when the column is made in a spiral shape the coiled tubing configuration is difficult to assemble, and connecting the ends of neighboring spiral tubing is rather difficult.

[0009] Another structure that can be used in a countercurrent chromatography column assembly comprises a plurality of separation disks having a plurality of spiral flow channels carved, etched, or molded on the surface of a first side of each separation disk as described in U.S. Patent Number 6,379,973, for example. The spiral flow channel has an inlet end and an outlet end, wherein fluid typically flows along the path of the spiral channel from the inlet end to the outlet end. The spiral channel of one separation disk can be serially connected to the spiral channel of another separation disk by stacking multiple separation disks adjacent to one another with a septum separating each pair. Preferably, an outlet end of a channel on one disk comiects to the inlet end of the channel on the next adjacent disk.

[0010] The flow channel design for countercurrent chromatography, however, has a number of disadvantages. The quality of the material comprising the separations disks is important for the accuracy of the instrument, and a high quality of machining of the channels is required so as not to allow fluid to leak from one channel to the next, and to ensure even flow of the fluid through the channels. The high quality required of the separation disk material and corresponding machining generally results in high manufacturing costs for the disks.

Summary of the Invention

[0011] hi one aspect of the invention, a disk shaped tube support for use in a countercurrent chromatography apparatus comprises a pair of opposed surfaces, an inner opening, an outer edge, at least one spiral groove formed in at least one of the surfaces and configured to accommodate at least one layer of fluid flow tubing, the at least one spiral groove having an inner groove portion and an outer groove portion, and at least one slot extending at least partially through the disk connecting the inner opening to the inner groove portion and the outer edge to the outer groove portion.

[0012] an additional aspect of the invention, the tube support comprises plastic, and more particularly, the tube support comprises a material chosen from the group consisting of foam plastic, polyethylene, and polypropylene.

[0013] The tube support may further comprise at least one return path formed in the tube support and configured to substantially accommodate a length of tubing from the outer groove portion of the at least one spiral groove to the inner groove portion of the same or a different spiral groove. [0014] In another aspect of the invention, the tube support comprises at least two interleaved spiral grooves, and may further comprise at least one return path formed in the tube support and configured to substantially accommodate a length of tubing from the outer groove portion of at least one of the interleaved spiral grooves to an inner groove portion of a different one of the interleaved spiral grooves.

[0015] In yet another aspect of the invention, a countercurrent chromatography apparatus comprises a disk having at least one spiral groove formed in a surface of the disk, and at least one layer of fluid flow tubing, positioned in the at least one spiral groove.

[0016] The apparatus can further comprise at least one radial return path formed in a portion of the disk, and at least one layer of tubing positioned in the at least one radial return path so as to provide a path between an outer groove portion of the at least one spiral groove and an inner groove portion of the same or a different spiral groove.

[0017] In an additional aspect of the invention, the fluid flow tubing has a circular cross-section, a rectangular cross-section, or a convoluted shape. Also, the fluid flow tubing may comprise polytetrafluoroethylene, and the disk may comprise a material chosen from the group consisting of foam plastic, polyethylene, and polypropylene

[0018] In yet another aspect of the invention, a method of manufacturing a plate assembly for use in high speed countercurrent chromatography comprises positioning at least one layer of fluid flow tubing in a spiral groove formed in a disk surface.

[0019] The method may further comprise positioning at least one layer of fluid flow tubing along a return path channel formed in a surface of the disk. The method may also comprise positioning at least one layer of fluid flow tubing in a slot formed in the disk, and positioning at least one layer of fluid flow tubing along a return path formed in a disk surface. In addition, the return path may be a groove, the fluid flow tubing can comprise polytetrafluoroethylene, and the disk may comprise plastic.

[0020] In yet another aspect of the invention, a disk shaped tube support for use in a countercurrent chromatography apparatus comprises a pair of opposed surfaces, an inner opening, an outer edge, a plurality of interleaved spiral grooves formed in at least one of the surfaces and configured to accommodate at least one layer of fluid flow tubing, each of the plurality of interleaved spiral grooves having an inner groove portion and an outer groove portion, and at least one slot extending at least partially through the disk connecting the inner opening to the inner groove portion of at least one of the plurality of interleaved spiral grooves and the outer edge to the outer groove portion of one of the plurality of interleaved spiral grooves.

[0021] hi an additional aspect of the invention, the tube support can comprise a material chosen from the group consisting of foam plastic, polyethylene, and polypropylene.

[0022] The tube support can further comprise at least one return path formed in the tube support and configured to substantially accommodate a length of tubing from the outer groove portion of the at least one spiral groove to the inner groove portion of the same or a different spiral groove. In addition, the at least one return path can be a groove formed in at least one of the surfaces of the disk.

Brief Description of the Drawings

[0023] Figure 1A is a top plan view of a tube support for use in high speed countercurrent chromatography.

[0024] Figure IB is a cross-sectional view of the tube support of Figure 1A taken along line A- A.

[0025] Figure 1C is a bottom plan view of the tube support of Figure 1 A.

[0026] Figure 2 A is a cross-sectional view of the tube support of Figures 1 A-C with multiple layers of tubing positioned therein.

[0027] Figure 2B is a partial bottom plan view of the tube support of Figure 2 A.

[0028] Figure 3 A is a top plan view of a tube support having a plurality of spiral grooves.

[0029] Figure 3B is a cross-sectional view of the tube support of Figure 3 A taken along line B-B.

[0030] Figure 3C is a bottom plan view of the tube support of Figures 3A-B.

Detailed Description of the Preferred Embodiment [0031] Embodiments of the invention will now be described with reference to the accompanying Figures, wherein like numerals refer to like elements throughout. The terminology used in the description presented herein is not intended to be interpreted in any limited or restrictive manner, simply because it is being utilized in conjunction with a detailed description of certain specific embodiments of the invention. Furthermore, embodiments of the invention may include several novel features, no single one of which is solely responsible for its desirable attributes or which is essential to practicing the inventions herein described.

[0032] Although embodiments of the invention have various applications, many advantageous embodiments of the present invention are directed to an improved plate apparatus for use in countercurrent chromatography. Applicable chromatography techniques include those using synchronous planetary motion such as X-type, -type, and I-type chromatography. The apparatus and methods described herein are especially advantageous when applied to high-speed countercurrent chromatography (HSCCC) with high flow rates. The plate design may also be employed in large column applications for industrial-scale separations of samples by mounting the column assembly on a slowly rotating horizontal shaft.

[0033] Some aspects of the invention are based, in part, on the fact that system cost and performance is improved when the column used in counter current chromatography is formed as a length of tubing which is installed within one or more grooves in a plate or disk (also referred to herein as a tube support).

[0034] One embodiment of a plate or disk shaped tube support 60 for use in a countercurrent chromatography apparatus is illustrated in Figures 1A-1C. Figure 1A is a top view of the tube support 60, Figure IB is a cross-sectional view of the tube support 60 taken along line A-A of Figure 1A, and Figure 1C is a bottom plan view of the tube support 60. The tube support 60 comprises a single spiral groove 62 carved, etched, or molded in an upper surface 63 of the tube support 60 and configured to accommodate or support a length of fluid flow tubing, as described further below. Although the spiral groove 62 is illustrated as having a rectangular cross-section, it will be appreciated that grooves having different geometrically shaped cross-sections may be used, such as a groove with an arcuate or semicircular cross-section. [0035] The spiral groove 62 originates at an inner end 64 located proximal to a center opening 66 of the tube support 60, and ends at an outer end 68 proximal to an outer rim 70 of the tube support 60. hi this embodiment, the inner end 64 and outer end 68 are located at substantially the same angular position on the tube support 60. A length of tubing having an outer diameter approximately equal to or slightly smaller than the width of the groove can then be positioned in the spiral groove 62 with a first end at the inner end 64 and a second end at the outer end 68 to provide a fluid flow path. The tubing can comprise a flexible fluid flow tubing material, a variety of which are well known in the art and are widely commercially available. In one advantageous embodiment, the tubing comprises polytetrafluoroethylene (PTFE).

[0036] By routing the fluid through tubing arranged on a grooved tube support rather than directly inside and in contact with the walls of the channels on a disk, the cost of the disk material and required precision of manufacturing can be significantly reduced, hi particular, the disk can comprise any solid material since the disk material itself is not exposed to the sample fluid. The tube support material is advantageously rigid, easily machined or molded, and inexpensive. Examples include polyethylene, polypropylene, and foam plastic. Also, the molding or machining precision required in forming the spiral channel or groove can be reduced where there is no concern regarding fluid leakage or concentration of particles at imperfection points along the groove.

[0037] In addition, leakage can occur in a chromatography apparatus which routes the fluid directly inside and in contact with spiral channels on the surface of a disk when a second disk or other sealing piece is positioned directly above the disk, and must be arranged flush with the channeled surface to avoid leakage of the fluid sample. In addition, when a column of disks are employed, the fluid connection path between neighboring disks must be precisely aligned such that fluid leakage or stoppage due to complete misalignment is avoided. The use of a single length of tubing for fluid flow avoids these leakage problems because the number of joints or connections present within each spiral and between different spiral layers can be minimized.

[0038] hi one advantageous embodiment, a single tube support can be configured to accommodate a continuous length of tubing to form several layers of spirals without connection joints, thereby avoiding leakage and contamination of the fluid sample undergoing the chromatography.

[0039] Thus, in one embodiment of the tube support, more than one layer of tubing is positioned in the spiral groove 62. To facilitate accommodation of multiple layers of tubing, a return path is provided in the tube support 60 for the tubing between the outer end 68 and inner end 64 of the spiral groove 62. hi one embodiment, an inner radial slot 71 is formed in the tube support 60 from the center opening 66 to an inner portion of the spiral groove 62, and an outer radial slot 72 is formed from an outer portion of the spiral groove 62 through the outer rim 70. The inner and outer radial slots 71, 72 can be fonned at the angular location of the inner end 64 and the outer end 68 of the spiral groove 62, as illustrated in Figures 1A-1C.

[0040] The inner and outer radial slots 71 „ 72 also provide a path for the tubing between the upper surface 63 and a radial return path groove, channel, or slot 73 formed in a lower surface 74 of the tube support 60 (Figure 1C). The inner end 64 and the outer end 68 of the spiral groove 62 correspond to the location of an inner end and an outer end of the return path groove 73, and the return path groove 73 is configured to accommodate at least one segment of tubing such that a segment of tubing between each spiral layer on the disk is housed in the return path groove 73.

[0041] Where multiple layers of tubing are housed in the spiral groove 62, the return path groove 73 can be configured to house a corresponding number of tubing segments wherein the return path groove 73 is configured with a depth such that multiple tubing segments can be accommodated in a layered configuration. Alternately, the return path groove 73 can be configured with a width such that multiple tubing segments can be arranged in a side-by-side configuration. In addition, the return path groove 63 may be configured to accommodate a plurality of tubing segments in both a layered and side-by-side configuration.

[0042] As illustrated in Figures 2A and 2B, a spiral column of tubing can be arranged on the tube support 60 by winding a desired length of tubing 80 onto a spool 82 which can freely pass through the center opening 66 of the tube support 60. Alternatively, the tubing may be bundled compactly to pass through the center opening 66. For a small disk with a small center hole, the tubing can be wound onto a long narrow spool, for example. [0043] The tubing 80 is wound around the spool 82 in a configuration such that as a first end of the tubing 80 is positioned at the imier end 64 of the spiral groove 62, the tubing 80 is unwound from the spool 82 and positioned along the spiral groove 62 by rotating the spool 82 in a direction opposite that of the spiral groove 62. For example, if the spiral groove 62 spirals in a clockwise direction from the inner end 64 to the outer end 68, the tubing 80 would be unwound from the spool 82 and positioned along the spiral groove 62 by rotating the spool 82 in a counter-clockwise direction.

[0044] When the tubing reaches the outer end 68 of the spiral groove 62, the tubing is passed through the outer slot 72 by passing the spool 82 around the outer rim 70 and under the tube support 60, and the tubing 80 is bent through the outer slot 72 and into the return path groove 73 and positioned in the return path groove 73. When the tubing has been sufficiently positioned along the return path groove 73, the spool 82 is passed through the center opening 66, and the tubing is bent through the inner slot 71 to return to the inner end 64 of the spiral groove 62.

[0045] Subsequent layers of tubing can be wound on the tube support 60 by pushing the previously positioned layer of tubing deeper into the spiral groove 62 and return path groove 73. Thereby, a plurality of spiral layers of tubing can be accommodated on a single tube support as shown in Figures 2A and 2B so as to form a spiral column of tubing on a single plate.

[0046] It will be appreciated by a person skilled in the art that the return path groove is not restricted to a radial or groove configuration, and other return path configurations are anticipated, such as a channel or slot having a first end and a second end at different angular positions.

[0047] The single spiral groove embodiment of the tube support 60 provides an asymmetrical distribution of the tubing and groove, which may require careful balancing of the column for centrifugation. However, a tube support having a plurality of spiral grooves can provide a more advantageous structure which does not require such careful balancing.

[0048] The column for counter current chromatography can also be fonned as a length of tubing which is installed within a plurality of grooves in a plate or disk, wherein the grooves are interleaved. The centrifugal force gradient produced by the spiral pitch in the interleaved groove embodiment of the tube support increases the efficiency of distribution of the heavier phase in the periphery and the lighter phase in the central portion of the column.

[0049] In one embodiment, multiple interleaved spiral grooves are formed symmetrically around the center of a plate or disk shaped tube support for use in a countercurrent chromatography apparatus, such that the spiral pitch is increased as compared to the spiral pitch of the single spiral groove embodiment of Figure 1A.

[0050] Figure 3 A is a top plan view of a tube support 100 having a plurality of interleaved spiral grooves configured to accommodate a plurality of layers of tubing, Figure 3B is a cross-sectional view of the tube support 100 taken along line B-B of Figure 3 A, and Figure 3C is a bottom plan view of the tube support of Figures 3A-B.

[0051] The tube support 100 comprises four interleaved spiral grooves 104, 106, 108, and 110 fonned in an upper surface 112 of the tube support 100. It will be appreciated, however, that the number of spiral grooves can vary, and the number of grooves illustrated and described herein is only exemplary in nature. Each spiral groove 104, 106, 108, 110 has an inner end denoted I_1; I₂, 1₃, and , respectively, located proximal to a center opening 120 of the tube support 100. Each groove 104, 106, 108, 110 spirals from its inner end I_ls I₂, 1₃, and J4 to an outer end denoted Oi, O₂, O₃, and O₄, respectively, located proximal to an outer rim 122 of the tube support 100. As discussed above with respect to the single spiral embodiment, the interleaved spiral grooves 104, 106, 108, 110 can have a cross-section other than the rectangular shape illustrated, such as an arcuate or semi-circular shape.

[0052] hi the embodiment of the tube support 100 illustrated in Figures 3A-C, the inner ends I , I₂, 1₃, and I₄ are positioned along an inner circumference of the tube support 100 at 90° intervals, and each spiral groove 104, 106, 108, and 110 forms 3.25 spiral turns such that the outer end (Oi, O , O , and O₄) of a given groove is located at substantially the same angular position as the inner end (I₂, I₃, I and If) of the next groove. Thus, as shown in Figure 3 A, O] is at the same angular orientation as I , O₂ is at the same angular orientation as I , etc.

[0053] Four pairs of inner and outer slots are formed in the tube support 100, comprising first inner and outer slots 130, 131, second inner and outer slots 132, 133, third inner and outer slots 134, 135, and fourth inner and outer slots 136, 137, wherein each pair of inner and outer slots are formed at substantially the same angular position on the tube support 100. The inner slots 130, 132, 134, 136 are formed from the center opening 120 to an inner portion of a corresponding spiral groove 106, 108, 110, 112, respectively, and the outer slots 131, 133, 135, 137 are formed from the outer rim 122 to an outer portion of the previous spiral groove 104, 106, 108, 110. Each pair of inner and outer slots 130, 131, 132, 133, 134, 135, 136 provides a path for the tubing between the upper surface 112 and a radial return path groove, channel, or slot 140, 142, 144, 146 (Figure 3C) formed in a lower surface 148 of the tube support 100. More specifically, the inner end I₂ and the outer end Oi correspond to the location of an inner end and an outer end of the return path groove 140, wherein the return path groove 140 is configured to accommodate at least one segment of tubing.

[0054] As previously discussed, it will be appreciated by a person skilled in the art that the return path is not restricted to a radial or groove configuration, and other return path configurations are anticipated.

[0055] Tubing can be positioned on the tube support 100 in a procedure similar to that previously described with respect to the tube support 60 having a single spiral groove. More specifically, starting at the first inner end I_l9 a length of tubing is unwound from a spool and positioned along the first spiral groove 104 to the outer end 0_\. The spool of tubing is then passed around the outer rim 122 and under the tube support and the tubing is bent through the first outer slot 131 and positioned along the first return path groove 140. When the tubing has been sufficiently positioned in the first return path groove 140, the spool is moved up through the center opening 120, and the tubing is through the first inner slot 130 and positioned at the inner end I₂ of the second spiral groove 106.

[0056] The process of positioning the tubing in the remaining three spiral grooves 106, 108, 110 and return path grooves 142, 144, 146 is performed similar to the process used to position the tubing along the first spiral groove 104 and first return path channel 140.

[0057] Similar to the tube support 60 with a single spiral groove, multiple layers of tubing can be arranged on the tube support 100 with four spiral grooves, wherein each time an additional layer of tubing is positioned along one of the spiral grooves 104, 106, 108, 110 or one of the return path grooves 140, 142, 144, 146, the previous layer of tubing is pushed deeper into the groove. Thereby, a plurality of spiral layers can be accommodated on a single, balanced tube support with increased spiral pitch as compared to a tube support with a single spiral groove.

[0058] Tubing with a circular cross section can be used to provide a fluid flow path on the tube support along the spiral and radial grooves, however, tubing having a cross- section with a non-circular geometry or convoluted shape can provide improved results. For example, tubing with a circular cross section has been found to produce a plug flow in tubing with a small diameter, particularly for the organic mobile phase of a two-phase solvent system with high interfacial tension and/or small density differences between the two phases. Such an effect may be largely reduced by implementing a tubing with a rectangular or triangular cross section, hi addition, tubing having a cross section with a rectangular or triangular cross section, for example, can provide improved stacking conditions for implementing multiple levels of spiral fluid flow paths on a single tube support. Alternately, tubing having a non-circular cross section can be twisted before positioning on the tube support so as to improve the partition efficiency.

[0059] In accordance with the foregoing, certain embodiments of the invention provide an improved plate design for use in high speed countercurrent chromatography. It will be appreciated, however, that no matter how detailed the foregoing appeai-s in text, the invention can be practiced in many ways. As is also stated above, it should be noted that the use of particular terminology when describing certain features or aspects of the invention should not be taken to imply that the terminology is being re-defined herein to be restricted to including any specific characteristics of the features or aspects of the invention with which that terminology is associated. The scope of the invention should therefore be construed in accordance with the appended claims and any equivalents thereof.

Claims

WHAT IS CLAIMED IS:

1. A disk shaped tube support for use in a countercurrent chromatography apparatus, comprising: a pair of opposed surfaces; an inner opening; an outer edge; at least one spiral groove formed in at least one of said surfaces and configured to accommodate at least one layer of fluid flow tubing, said at least one spiral groove having an inner groove portion and an outer groove portion; and at least one slot extending at least partially through said disk connecting said inner opening to said inner groove portion and said outer edge to said outer groove portion.

2. The tube support of Claim 1, wherein said tube support comprises plastic.

3. The tube support of Claim 1, wherein said tube support comprises a material chosen from the group consisting of foam plastic, polyethylene, and polypropylene.

4. The tube support of Claim 1, further comprising at least one return path formed in said tube support and configured to substantially accommodate a length of tubing from said outer groove portion of said at least one spiral groove to the inner groove portion of the same or a different spiral groove.

5. The tube support of Claim 4, wherein said return path is a groove formed in one of said surfaces.

6. The tube support of Claim 1, comprising at least two interleaved spiral grooves.

7. The tube support of Claim 6, further comprising at least one return path formed in said tube support and configured to substantially accommodate a length of tubing from the outer groove portion of at least one of said interleaved spiral grooves to an inner groove portion of a different one of said interleaved spiral grooves.

8. A countercurrent chromatography apparatus comprising: a disk having at least one spiral groove formed in a surface of said disk; and at least one layer of fluid flow tubing positioned in said at least one spiral groove.

9. The apparatus of Claim 8, further comprising at least one radial return path formed in a portion of said disk, and at least one layer of tubing positioned in said at least one radial return path so as to provide a path between an outer groove portion of said at least one spiral groove and an inner groove portion of the same or a different spiral groove.

10. The apparatus of Claim 9, wherein said radial return path is a groove formed in a surface of said disk.

11. The apparatus of Claim 8, wherein said fluid flow tubing has a circular cross- section.

12. The apparatus of Claim 8, wherein said fluid flow tubing has a rectangular cross-section.

13. The apparatus of Claim 8, wherein said fluid flow tubing has a convoluted shape.

14. The apparatus of Claim 8, wherein said fluid flow tubing comprises polytetrafluoroethylene.

15. The apparatus of Claim 8, wherein said disk comprises a material chosen from the group consisting of foam plastic, polyethylene, and polypropylene.

16. A method of manufacturing a plate assembly for use in high speed countercurrent chromatography comprising positioning at least one layer of fluid flow tubing in a spiral groove formed in a disk surface.

17. The method of Claim 16, further comprising positioning at least one layer of fluid flow tubing in a slot formed in said disk.

18. The method of Claim 17, further comprising positioning at least one layer of fluid flow tubing along a return path formed in a disk surface.

19. The method of Claim 16, further comprising positioning at least one layer of fluid flow tubing along a return path formed in a disk surface.

20. The method of Claim 19, wherein said return path is a groove.

21. The method of Claim 16, wherein said fluid flow tubing comprises polytetrafluoroethylene.

22. The method of Claim 16, wherein said disk comprises plastic.

23. The method of Claim 16, wherein said disk comprises a material chosen from the group consisting of foam plastic, polyethylene, and polypropylene.

24. A disk shaped tube support for use in a countercurrent chromatography apparatus, comprising: a pair of opposed surfaces; an inner opening; an outer edge; a plurality of interleaved spiral grooves fonned in at least one of said surfaces and configured to accommodate at least one layer of fluid flow tubing, each of said plurality of interleaved spiral grooves having an inner groove portion and an outer groove portion; and at least one slot extending at least partially through said disk connecting said inner opening to said inner groove portion of at least one of said plurality of interleaved spiral grooves and said outer edge to said outer groove portion of one of said plurality of interleaved spiral grooves.

25. The tube support of Claim 24, wherein said tube support comprises a material chosen from the group consisting of foam plastic, polyethylene, and polypropylene.

26. The tube support of Claim 24, further comprising at least one return path formed in said tube support and configured to substantially accommodate a length of tubing from said outer groove portion of said at least one spiral groove to the inner groove portion of the same or a different spiral groove.

27. The tube support of Claim 26, wherein said at least one return path is a groove formed in at least one of said surfaces.