WO1992004958A1

WO1992004958A1 - Interchangeable modular chromatography column with self-sealing connectors

Info

Publication number: WO1992004958A1
Application number: PCT/EP1991/001781
Authority: WO
Inventors: Kjell Evert Karlsson; Lennart SÖDERBERG
Original assignee: Dynochrom As
Priority date: 1990-09-17
Filing date: 1991-09-17
Publication date: 1992-04-02
Also published as: GB9020306D0

Abstract

A modular chromatography column unit (10) adapted to cooperate with a holder (21) therefor, the ends of said column (1) being fitted with self-sealing connector means (15, 16) which are slideably engageable and cooperable with corresponding self-sealing connector means (17, 18) on said holder (21) to provide a fluid-tight flow path through each pair of cooperating connector means (15, 17 and 16, 18).

Description

INTERCHANGEABLE MODULAR CHROMATOGRAPHYCOLUMN WITH SELF-SEAL¬ ING CONNECTORS This invention relates to an improved form of interchangeable chromatography column, more particularly to novel modular chromatography columns of particular use in high performance liquid chromatography (HPLC) . One aspect of the growth of HPLC techniques has been the development of a wide range of interchangeable chromatography columns in cartridge form. Such columns may, for example, be made of a metal such as stainless steel or titanium or may employ a plastic (e.g. polyethylene) column which, by virtue of the operating pressures which may be encountered, will normally be enclosed and compressed by a metal cartridge holder.

In view of these relatively high operating pressures it has hitherto been thought necessary to use screw connectors for all connections to such cartridges. Since the dimensions of existing cartridges vary widely depending on, for example, the intended use and the nature of the support material, there has been an inevitable proliferation of connecting pieces, capillary receiving pieces, reducing pieces, adaptors etc., together with their associated nuts, ferrules and the like, in order to incorporate particular cartridges into specific analytical or preparative systems. This can create considerable practical difficulties when, for example, it is desired to change a particular chromatography technique or column size or to adapt to multiuser operation of apparatus, especially when automated systems are concerned.

The present invention is based on our finding that substantial benefits can accrue if chromatography columns are produced in a standard modular unit form, permitting their use with permanently fixed standard holders. Thus according to one aspect of the present invention there is provided a modular chromatography column unit adapted to cooperate with a holder therefor, the ends of said column being fitted with self-sealing connector means which are slideably engageable and cooperable with corresponding self-sealing connector means on said holder to provide a fluid-tight flow path through each pair of cooperating connector means.

The self-sealing connector means may, for example, contain slide valves which operate to close off the apertures of the column and holder when these are not connected together, to prevent ingress of contaminants and egress of reagents. Such valves, which may if appropriate be spring loaded, are positioned such that they are displaced upon slideable engagement of each cooperating pair of connector means so as to permit fluid flow between the holder and column. Appropriate slide valves and coupling units are described in, for example, U.S. Patent No. 4,576,199 (the contents of which are incorporated herein by reference) although only with reference to surgical tubing, for example in infusion or transfusion of body fluids or peritoneal dialysis.

In order to permit easy visual examination of the column and its contents while also achieving maximum strength of construction, it is advantageous to construct the column from glass or a substantially transparent plastics material (e.g. polyethylene or polymethylmethacrylate) and to enclose the column in an essentially rigid, clear or substantially completely clear plastics material (e.g. clear polymethacrylate or epoxy resin) . This enclosing material is preferably in the form of a block to ensure ease of handling and to give a solid, eye-appealing structure which can easily be aligned with a correspondingly shaped holder.

The holder may, for example, possess a pair of extending arms carrying appropriate connector means adapted to engage corresponding connector means located at opposite ends of a chromatography column module. Appropriate grooves, tongues, recesses or the like may optionally be provided on the arms and/or the ends of the modules to assist positive location of a module while being fitted to the holder, thereby facilitating cooperation of the slideably engageable connectors.

Alternatively the columns may possess parallel, radially extending feed arms at each extremity, such arms preferably being positioned within any enclosing material such as a block and being engageable with corresponding socket connectors in one side of the holder.

The holder will normally be permanently positioned and incorporate fixed connections to one or more appropriate systems for input and efflux of fluids, e.g. such liquids as buffer eluants and liquid samples, particularly via automatic analysis devices. Rapid interchange of column modules may thus be effected simply by "plugging in" each module to the holder.

The holder is advantageously fitted with a sample inlet port whereby uniform, reproducible injection of samples to the heads of the modular chromatographic columns may readily be achieved. This port may, for example, be bored into the input fluid flow path in a region closely adjacent to the connector means of the appropriate arm of a holder, and when not in use may be closed by, for example, a screw cap or insert.

It may be advantageous, particularly where use of substantially transparent columns and enclosing material is envisaged, to incorporate a light into the body of the holder to facilitate viewing of the column and its contents. Such a light may, for example, be controlled by switching means such as microswitches operated when a chromatography column module is inserted into the holder. It is particularly preferred to arrange the switching means in such a way that the light operates in different modes (e.g. either continuous or flashing) depending on which way round the module is inserted, e.g. as a precaution against inadvertent reverse flow of fluid through a column being permitted or to ensure the correct positioning of a column for cleaning protocols. The chromatography columns of modules according to the invention may contain any convenient chromatographic support material, for example any of the variety of microparticulate materials proposed for use in HPLC. The use of monodisperse polymer particles, e.g. prepared as described in U.S. Patent No. 4,459,378 (the contents of which are incorporated herein by reference) may be particularly beneficial. Thus, for example, highly onosized particles of polymers such as polystyrene crosslinked with divinylbenzene and functionalised with acid groups such as sulphonic or carboxylic acid groups, with base groups such as tertiary amine or quaternary ammonium hydroxide groups or with salts of such acid and base groups may permit especially high resolution of proteinaceous materials in ion exchange separations.

The low operating pressures that the use of monodisperse particles permit is also advantageous and render them compatible with the use of peristaltic pumps.

When using columns containing microparticulate chromatographic support material it will normally be necessary to provide filter material (e.g. a conventional filter pad) at each end of the support material in a column in order to retain the support material in position. It has been found advantageous, especially when using monodisperse polymer particles, to position a distribution plate between the filter and the support material at each end of the latter, to enhance substantially uniform fluid flow through the support material. Such a distribution plate may, for example, possess a plurality of concentric annular grooves on the face contacting the support material, said grooves communicating via one or more (preferably symmetrically disposed) radial grooves with an axial bore through the plate. The depths and pattern of the grooves may be selected to maximise the uniformity of fluid flow for any particular column cross-sectional area. The filter and distribution plate may for example be fixably located across a tubular member adapted to be sealingly inserted into one end of the chromatography column, said tubular member extending to or otherwise carrying the connector means for that end of the column. In a finished modular assembly according to a preferred feature of this embodiment of the invention the tubular member/connector means is conveniently locked in its operating position by one or more screws or other locating (e.g. snap-fit) means passing through or otherwise cooperating with the block or other material surrounding the column.

In order to facilitate filling of support material into chromatography columns according to this last- mentioned embodiment of the invention, a filling orifice is advantageously provided in the side of the column towards one end thereof, such that said orifice is sealed by the appropriate tubular member in its operating position but permits introduction of support material (e.g. via a side arm and/or bore in the surrounding block or other material) if the tubular member is in a partially withdrawn position. For ease of manufacture and operation it may be preferred to construct such filling orifices and any associated side arms and/or bores at both ends of the column, e.g. to give a symmetrical assembly, although in practice only one filling orifice will normally be used. It will be understood that the tubular member should still close the end of the column in the partially withdrawn position referred to above. The microparticulate support material, e.g. comprising monodisperse polymer particles, is conveniently introduced in the form of a slurry in an appropriate liquid, e.g. water, preferably through the upper orifice of a substantially vertically disposed column. The pressure of the incoming slurry flow is desirably maintained at a substantially constant level throughout the filling procedure, an equal pressure of pure liquid also, if necessary or desired, being applied via the connector means of the partially withdrawn tubular member to prevent any backflow of liquid. Liquid is allowed to pass through the connector means of the opposite tubular member, whereby a bed of support material builds up above the filter/distribution plate of said opposite tubular member. Once a desired level of support material has entered the column the partially withdrawn tubular member is moved to and locked in its operating position, fixing the column of support material between the opposing filter/distribution plates, the applied slurry and liquid pressures being maintained until this operation is complete.

The application of constant slurry and liquid pressures in the filling stage encourages substantially completely uniform packing density for the support material throughout the entire volume of the column. By selecting a filling pressure which exceeds the maximum intended working pressure for the column in subsequent use it is possible to avoid the formation of liquid pockets, e.g in the upper part of the column, in operational usage within the specified pressure range.

This method of filling chromatography columns, which may also be applied to apparatus other than the modular columns hereinbefore described (e.g. omitting the self-sealing connector means) and apparatus therefor constitute further features of the present invention.

The invention is now further described with reference to the accompanying non-limitative drawings, in which:-

Fig. 1 represents a schematic side elevation of a pair of cooperable self-sealing connector means of use - 1 - in apparatus according to the invention;

Fig. 2 represents a schematic side elevation of the connector means of Fig. 1 in slideably engaged position; Fig. 3 represents a schematic side elevation of a holder and a chromatography column module according to the invention;

Fig. 4 represents a vertical cross-section through the column and encasing block of a module according to the invention; Fig. 5 represents a plan view of one face of a distribution plate of use in columns according to the invention;

Fig. 6 represents a schematic side elevation of a module in which the column is ready to be filled with support medium; and

Fig. 7 represents a schematic side elevation of the finished assembly of the module of Fig. 6 after filling of the column with support material.

In the apparatus of Fig. 1 chromatography column 1 is encased in block 2 and is closed at its upper end by slide 3 and rubber seal 4. Slide 3 is maintained against a stop (not shown) by pressure from spring 5. Fluid feed duct 6 of a holder arm (not shown) is similarly closed by slide 7, pressured by spring 8. Rubber seal 9 is positioned below said slide 7.

In the engaged position shown in Fig. 2, the two units are held together by, for example, cooperating flanges and recesses (not shown) such that slides 3 and 7 are displaced laterally, compressing springs 5 and 8 and permitting fluid flow between feed duct 6 and column 1 via cooperating seals 4 and 9.

As may be seen from Fig. 3, a module 10 may conveniently comprise a column 1 encased in a block 2 and having filling arms and orifices 11 and 12 sealed respectively by tubular members 13 and 14 which contain filter/distribution plate assemblies (not shown) and terminate respectively in connector means 15 and 16. These connector means 15 and 16, which are preferably female, are engageable with (preferably male) connector means 17 and 18 on upper and lower arms 19 and 20 of column holder 21 and may be latched in the engaged position by, for example, lock mechanisms 22 and 23 or a corresponding snap-fit mechanism, thereby permitting fluid to flow from source 6 through column 1 to fluid exit 24.

In the embodiment shown in Fig. 4 the overall column comprises a glass tube 25 set in block 2 and extended at each end by cylindrical recesses 26 and 27 moulded into the block 2 and communicating with filling arms 11 and 12.

Insertion of tubular members (not shown) into recesses 26 and 27 will effectively seal off filling arms 11 and 12 and define the operating length of the column as being the glass tube 25.

As shown in Fig. 5, that face of the distribution plate 28 which contacts the support material conveniently comprises a plurality of annular grooves 29 connected to central orifice 30 by radially extending grooves 31.

Fig. 6 demonstrates the manner in which the upper tubular member/connector means assembly 13/15 may be partially withdrawn to permit a slurry of support material to be fed into column 1 through side arm 11 while simultaneously applying an equal pressure of liquid to connector means 15 and permitting liquid to escape via connector means 16. As shown in Fig. 7 the upper tubular member/connector means assembly 13/15 may thereafter be lowered and locked in its operating position to seal off side arm 11 and fixably to locate the support material 32 between the filter/distribution plate shown at 33 in upper tubular member 13 and the corresponding filter/distribution plate of lower tubular member 14 (not shown) .

Claims

1. A modular chromatography column unit (10) adapted to cooperate with a holder (21) therefor, the ends of said column (1) being fitted with self-sealing connector means (15,16) which are slideably engageable and cooperable with corresponding self-sealing connector means (17,18) on said holder (21) to provide a fluid- tight flow path through each pair of cooperating connector means (15,17 and 16,18) .

2. A modular unit (10) as claimed in claim 1 wherein each pair of cooperating connector means (15,17 and 16,18) comprise two spring loaded slide valves (3,5 and 7,8) positioned such that they are mutually displaced upon slideable engagement of the module (10) into a holder (21) so as to permit fluid flow between the holder (21) and the column (1) .

3. A modular unit (10) as claimed in claim 1 or claim 2 wherein the column (1) is constructed from glass or a substantially transparent plastics material and is enclosed in an essentially rigid clear or substantially completely clear plastics material (2) .

4. A modular unit (10) as claimed in claim 3 wherein the enclosing plastics material (2) is in the form of a block.

5. A modular unit (10) as claimed in any of the preceding claims in association with said holder (21) .

6. A modular unit (10) as claimed in claim 5 in association with a holder (21) which possesses extending arms (19,20) carrying connector means (17,18) adapted to engage corresponding connector means (15,16) located at opposite ends of the module (10) .

7. A modular unit (10) as claimed in claim 5 or claim 6 in association with a holder (21) fitted with a sample inlet port to permit injection of samples to the head of the chromatography column (1) .

8. A modular unit (10) as claimed in any of claims 5 to 7 in association with a holder (21) which incorporates a light.

9. A modular unit (10) as claimed in claim 8 in association with a holder (21) wherein the light is controlled by switching means operated when the module (10) is inserted into the holder, in such a way that the light operates in different modes depending on the direction of insertion of the module (10) .

10. A modular unit (10) as claimed in any of the preceding claims wherein the chromatography column (1) contains microparticulate chromatographic support material (32) .

11. A modular unit (10) as claimed in claim 10 wherein the support material (32) comprises monodisperse polymer particles.

12. A modular unit (10) as claimed in claim 10 or claim 11 wherein the support material (32) is retained in position by filter material positioned at each end of the support material (32) .

13. A modular unit (10) as claimed in claim 12 wherein a distribution plate (28) is positioned between the filter material and the support material (32) at each end of the support material (32) .

14. A modular unit (10) as claimed in claim 13 wherein each distribution plate (28) possesses a plurality of concentric annular grooves (29) on the face contacting the support material (32) . said grooves (29) communicating via one or more radial grooves (31) with an axial bore (30) through the plate (28) .

15. A modular unit (10) as claimed in any of claims 12 to 14 wherein each filter means and any associated distribution plate (28) are fixably located across a tubular member (13,14) sealingly inserted into one end of the chromatography column (1) and extending to or otherwise carrying the connection means (15,16) for that end of the column (1) .

16. A modular unit (10) as claimed in claim 15 wherein at least one filling orifice (11,12) is provided in the side of the chromatography column (1) such that it is sealed by the appropriate tubular member (13,14) in its operating position but permits the introduction of support material (32) if the tubular member (13,14) is in a partially withdrawn position.

17. A modular unit (10) as claimed in claim 16 wherein the chromatography column (1) has been filled with support material (32) by applying a slurry thereof in an appropriate liquid at substantially constant pressure to a filling orifice (11) of a substantially vertically disposed column (1) , which orifice is open by virtue of the upper tubular member (13) being in a partially withdrawn position, if necessary or desired applying an equal pressure of liquid to the connector means (15) of said upper tubular member to prevent backflow of liquid, allowing liquid to pass through the connector means (16) of the lower tubular member (14) whereby a bed of support material (32) builds up above the filter means and any associated distribution plate (28) of said lower tubular member (14) , moving said upper tubular member (13) into its operating position once a desired level of support material (32) has entered the column (1) , and releasing the applied pressure of slurry and that of any liquid employed.

18. A modular unit (10) as claimed in claim 17 wherein the chromatography column (1) has been filled using slurry at a pressure exceeding the maximum intended working pressure for the column (1) .