Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/60—Construction of the column
  • G01N30/6004—Construction of the column end pieces
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
  • B01D15/08—Selective adsorption, e.g. chromatography
  • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
  • B01D15/20—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
  • B01D15/08—Selective adsorption, e.g. chromatography
  • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
  • B01D15/20—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
  • B01D15/206—Packing or coating
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/50—Conditioning of the sorbent material or stationary liquid
  • G01N30/56—Packing methods or coating methods
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
  • B01D—SEPARATION
  • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
  • B01D15/08—Selective adsorption, e.g. chromatography
  • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
  • B01D15/22—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the construction of the column
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/50—Conditioning of the sorbent material or stationary liquid
  • G01N30/56—Packing methods or coating methods
  • G01N2030/562—Packing methods or coating methods packing
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
  • G01N30/02—Column chromatography
  • G01N30/60—Construction of the column
  • G01N30/6004—Construction of the column end pieces
  • G01N30/6021—Adjustable pistons

Definitions

• the present invention relates to a new high performance preparative chromatography device which allows to combine a compression of the chromatographic bed and a use of the chromatography column.
• a chromatographic separation process the efficiency of the column used is a key parameter.
• the column is generally filled with a solid product called a stationary phase in the form of very fine grains usually of a size between 5 and 100 ⁇ m for high performance chromatography systems, forming the chromatographic bed.
• the arrangement of the grains inside the column must be as homogeneous as possible and moreover, it is necessary to avoid empty volumes between the chromatographic bed and the entry of the column.
• the columns with Dynamic Axial Compression have a piston or other sliding body driven by a jack actuated for example by an independent hydraulic circuit; the sliding body makes it possible to pack the packing in a chamber between this piston and a fixed bottom or removable cover. The sliding body comes into contact with the solid phase to maintain a constant mechanical pressure on the bed.
• DAC Dynamic Axial Compression
• French patent application FR-A-2 219 797 discloses a chromatography apparatus in the form of a column intended to contain a bed of adsorbent material.
• the apparatus consists of a tube comprising a cover and a body sliding in the tube for exerting pressure in the tube.
• the sliding body is a piston having at its head a porous plate permeable to fluids called frit.
• a suspension of particles of a material capable of constituting an adsorbent bed is introduced into the tube without its lid or its piston.
• the lid is fitted to the tube and then pressure is exerted on the suspension by means of the piston or sliding body.
• the liquid is forced through the porous plates, and said particles are compressed between the piston and the lid.
• This apparatus makes it possible to achieve, in an exemplary embodiment, the stage of compression of the chosen bed and the compaction of the desired particles when the sliding body has generally traveled between 1/4 and 3/4 of the height of the column, a function of the density of the suspension used to fill the column.
• this DAC system by using dedicated compression module.
• These modular systems comprise (i) an axial compression module dedicated to compressing the suspension into a bed, and (ii) a chromatography column equipped with a piston making it possible to ensure homogeneous filling of the column by axial compression at filling, compression which is however not maintained on the bed during the use of the chromatographic column, unlike columns DAC.
• the Merck Selfpacker system for which the filling protocol is described below.
• the column must be mounted with the piston via screws on the hydraulic compression system. Once the top flange of the column is removed, a fill tank is attached above the column. The suspension is introduced from the top into the tank and the column, the system is kept open. A vacuum is applied for 5 to 45 minutes on the line collected by the outlet of the lower flange of the column to cause formation of the grain bed by suction of the liquid through the column outlet frit (at the lower level) . The tank is then removed, the column closed by a flanged system which is fastened with screws, solvent is optionally injected, and the bed is then pressurized by moving the piston from bottom to top in the column. . After compression, the column is disconnected from the hydraulic system after locking the piston by a threaded rod. Applying a vacuum is a very time consuming operation.
• the Modcol system is also known, in which the compression is done this time from top to bottom, but without prior application of vacuum.
• the column lengths are necessarily larger than in the aforementioned case of Merck, since the columns must contain the suspension before compression.
• the piston is blocked by a threaded rod.
• a mechanical spring system can be used to maintain compression on the bed.
• Modcol technology uses elements that must be screwed together.
• the Amicon system uses a suspension loading in a system comprising a column and an adjustment unit, this unit serving in particular as a reservoir for loading the suspension of particles. After loading the suspension, an adapter is attached to the body of the column, which adapter contains the piston, and the compression of the bed can be done in a press, down. Then, the pressure is removed, the adapter is removed and the piston held in place by a safety system in the adjustment unit which is not removable and remains secured to the column.
• the Axxial system uses two opposing pistons that compress the bed in the column at the center of the system.
• the use of two pistons (without the column can not be recompressed next) is complex.
• JP 61283278 discloses a method for compressing a filler to obtain a chromatographic bed.
• Two lower and upper columns are used end to end, a piston being located at the free end of each column.
• the load is disposed in the lower column and the piston pushes the load towards the upper column; then the lower column is removed and a flange is disposed at the lower end of the upper column.
• the piston of the lower column does not enter the upper column.
• the piston of the upper column is actuated to compact the load against the flange.
• This document has the disadvantage of requiring two pistons, one of the pistons to transfer the load from the lower column to the upper column, the other of the pistons to pack the load in the upper column.
• the withdrawal of the lower column to place the flange is hazardous; indeed, the lower end of the load is found in the open air time to place the flange. This process is therefore not efficient.
• JP 63288138 discloses a method for compressing a filler to obtain a chromatographic bed.
• a piston is actuated to compact a load in a column; a cone between the piston and the column makes it possible to adapt the size of the piston to the diameter of the column, the diameter of the column being smaller than the diameter of the piston. It should be noted that the piston can not enter the column as long as the diameter of the piston is greater than the diameter of the column.
• the present invention relates to a modular chromatography device.
• the invention therefore provides a chromatography device comprising:
• a removable reservoir for a bed suspension a base comprising a jack adapted to a piston, said piston sliding in the removable reservoir and the column for compressing the suspension of solid grains in a chromatographic bed,
• the columns, reservoir and base are connected from top to bottom.
• the piston is provided with piston seal friction in the column and in the reservoir.
• the piston and the jack are connected by a coupling cylinder and possibly a set of anti-rotation washers abutting the piston reversibly.
• the columns, reservoir and base are secured at the flanges by means of fasteners by clamps.
• the base comprises two chambers receiving a hydraulic fluid under pressure, disposed on either side of the lower part of the jack.
• the column further comprises a set threaded rod and threaded flange receiving the threaded rod, said threaded rod adapting to the piston, this assembly replacing the reservoir when using the column as a column chromatography.
• the invention further relates to a chromatography column comprising a threaded rod and threaded flange assembly receiving the threaded rod, said threaded rod adapting to the piston, this assembly being fixed to the column by a fastening clamp.
• the invention also relates to a chromatography column comprising a threaded rod and threaded flange assembly receiving the threaded rod, said threaded rod adapting to the piston, this assembly being attached to the column, and wherein the threads of the rod and the tap are covered with an anti-seizing coating.
• the subject of the invention is also a method for compressing a chromatographic bed in a chromatography column, comprising the following steps:
• the method further comprises the following step:
• the method further comprises the following step:
• the method further comprises the step of recompression by clamping tightening the threaded rod.
• the invention further relates to a process for separating at least two compounds from a fluid to be chromatographed comprising chromatography of said fluid in the column according to the invention.
• the invention also relates to a process for separating at least two compounds from a fluid to be chromatographed comprising the bed compression steps according to the invention, and the flow of the fluid to be chromatographed through the bed and its collection. .
• the threads of the rod and the tap are covered with an anti-seizing coating.
• the end of the rod can be machined in hexagon to receive a wrench.
• a spring is present between said threaded rod and the piston.
• the device comprises a modular system shown in the right part of the figure, while the hydraulic system is shown on the left, a dashed line schematically separating the two sets.
• the hydraulic system comprises a pressure regulator 1, a hydropneumatic pump 2, a pressure gauge 3 and a reservoir 4 of hydraulic fluid, typically oil.
• the tank 4 is provided with a vent 5 and a discharge 18 for maintenance of the oil.
• a manual valve 6 is used to direct the oil to the modular system, as will be indicated below.
• a safety valve 7 and an emergency stop button 23 complete the hydraulic system.
• the modular system comprises, during the compression phase of the suspension in a chromatographic bed, the following elements: a column 10 closed on the upper part by a flange 15 held by a clamp 21a, a removable reservoir 9 intended to receive the suspension for chromatographic bed, and a base 8.
• a jack 14 is disposed in the base. It is adapted to a column piston 11 via a coupling cylinder 19.
• the coupling cylinder 19 is not integral with the column piston 1 1, but it transmits the pressure of the cylinder 14 to the piston 1 1 which can thus mount in the tank 9 and the column 10 and compress the bed or down to release the pressure on the piston 1 1 according to the position of the valve 6.
• Two chambers for hydraulic fluid 16 and 17 are disposed on either side of the cylinder 14, the cylinder sliding in the base between the two chambers by a seal 13.
• the chambers 16 and 17 are defined by the movement of the base 20 of the cylinder 14.
• the two chambers are kept separate and sealed together by the cylinder 14 and the gasket of the cylinder 13.
• the hydraulic system is used to pump the hydraulic fluid from one of these chambers and fill the second to move the cylinder up or down depending on the direction of pumping selected using the manual valve 6.
• the upper flange of the column is removed by removing the tie clip 21a and the column piston 11 is positioned at the base of the reservoir 9, the cylinder 14 being retracted downwardly to obtain a maximum available volume corresponding to the volume of the column 10 and the tank 9 into which the suspension is poured.
• the piston seals 12 make it possible to seal the walls and to maintain the suspension in the volume corresponding to the reservoir 9 and to the column 10.
• Variable reservoir lengths make it possible to accommodate variable suspension volumes.
• the cylinder 14 For the compression of the bed (operation called "packing" of the column), the cylinder 14 is pushed by the hydraulic fluid compressed by the hydropneumatic pump 2.
• the hydropneumatic pump 2 directs the hydraulic fluid, through a choice of the position of the valve 6, to the chamber 16, which leads to an upward movement of the cylinder 14.
• the reduced volume of the chamber 17 leads to a drain of the fluid which is returned to the tank 4.
• the difference in air volume that results in the tank 4 is compensated by the vent 5 which allows the air to escape or enter in order to maintain the atmospheric pressure in the tank 4.
• a coupling cylinder 19 is mounted fixed on the cylinder (for example by screwing) and receives at its upper part the piston 11.
• the piston originally placed on the coupling cylinder 19 pushes the suspension upwards to form the bed by compression.
• the piston is slid into the reservoir into the column.
• the piston 1 1 is not attached to the coupling cylinder 19.
• the piston 11 comprises seals 12 which prevent leakage of the container from the column and the reservoir downwards. Column lengths and / or variable reservoirs make it possible to accommodate variable bed heights.
• Clamps 21a, 21b, 21c are used to ensure the connection of the column and the other elements of the modular system. Clamps 21a and 21b make it possible to seal respectively between the upper flange and the column and between the column and the tank by means of O-rings placed between these elements.
• the speed of the compression of the bed is controlled in particular by the air flow of the regulator 1 supplying the hydropneumatic pump 2.
• the final pressure exerted on the bed is controlled by the air pressure fixed at the regulator which sets the pressure of the pump 2 and the ratio of the surfaces between the piston 1 1 and the lower part 20 of the cylinder 14.
• the safety valve 7 limits the pressure exerted on the compressed medium forming the bed. This valve is calibrated at a hydraulic pressure equal to the maximum operating pressure of the medium forming the bed multiplied by the ratio of the surfaces between the piston 1 1 and the lower part 20 of the cylinder 14.
• the cylinder is retractable, but it is not telescopic. It is not formed of multiple elements sliding into each other to gain space once retracted. Indeed, space saving is not necessary with the jack in the system according to the invention.
• the hydraulic fluid pressure is therefore applied to the same surface of the jack during the entire compression phase, this which makes it possible to maintain a constant force exerted on the piston and to have only one safety valve calibrated with a single pressure.
• the flange 15 is disengaged from the column 10 by removing the clamp 21a.
• the hydropneumatic pump 2 is actuated by an air inlet controlled by the pressure regulator 1 by positioning the valve 6 so that the hydraulic fluid enters the chamber 16, so as to discharge the fluid from the chamber 17 to the reservoir.
• the seed bed is extruded from the column 10 is recovered from the top of the column, for example in a container positioned at the column outlet.
• the jack 14 After having compressed the bed in the column 10, the jack 14 is retracted, and thus also the coupling cylinder 19 by turning the valve 6 so that the hydraulic fluid enters this time into the chamber 17, so as to repress the fluid from chamber 16 to the tank.
• the piston 11 remains in place by the effect of the friction forces exerted on the walls of the column 10.
• it could also be used a mechanical locking system, if desired.
• the bed is maintained between the piston and the flange. Due to the piston present in the column, the advantage is that only one piston is used both to place the chromatographic bed and then to adjust the pressure of the bed during a chromatographic separation operation.
• a spring for example of the Belleville washer type 22 (FIG 2b) can be arranged between the piston 1 1 and the threaded rod 24, so as to compensate for a possible pressure drop with time during the chromatography operations.
• the use of an intermediate removable reservoir 9 makes it possible to avoid the use of very long columns (as in particular in the case of Modcol). Chromatography columns shorter and therefore lighter are more easily manipulated. Since the reservoir can be available in several lengths, greater flexibility is achieved. Investment costs are also reduced.
• the intermediate tank 9 which is disconnected when using the column 10 thus allows a shorter column.
• the column in the invention is unitary, namely it comprises a single cylinder in which the piston slides and in which the chromatographic bed is located; there are no elements mounted successively on each other.
• the column 10 and the intermediate removable tank 9 are advantageously made of stainless metal, stainless steel and PTFE seals, so that the parts of the device in contact with the solvents or the products are chemically stable and are compatible with the requirements in this respect. especially for pharmacy.
• the internal diameters can typically vary from 20 to 200 millimeters, for column lengths of 50 to 600 millimeters, and reservoir lengths of 100 to 600 millimeters.
• the devices according to the invention using columns of larger diameters will incorporate preferably handling assistance means. The preferred use of fastening by clamps is very easy and very fast.
• the joint use of the anti-rotation assembly makes it possible to limit the drive of the piston by rotation and thus to control the pressure exerted on the compressed bed by the torque to be exerted on the threaded rod.
• the invention also offers an advantage by controlling the pressure exerted on the bed as a function of the tightening torque by the clamping nut 26 located at the base of the threaded rod 24.
• the fact of using a coating avoiding the seizing makes it possible to control the pressure exerted by the screw system, advantageously linearly. It is thus possible to manually recompress the column if necessary, by tightening the threaded rod.
• This technical feature is furthermore independent of the general system of the invention and may apply to any type of column. It is also possible to use an external hexagonal machining at the screw for clamping the piston on the bed. It is also possible to install an anti-rotation washer between the piston and the rod in addition to the jack.
• the end of the piston rod 24 is machined in the form of clamping nut 26 (previously described in connection with Figure 2a) to receive a key; this key can be used to manually tighten the piston to compensate for the eventual compression loss over time during chromatography operations; in addition, the use of a torque wrench makes it possible to dose the tightening according to the characteristics of the bed.
• clamping nut 26 previously described in connection with Figure 2a
• a nut with a flange machined in the form of a clamping nut 27 and inserted between the threaded flange 25 and the chromatography column 10; this nut slides on the base of the threaded flange 25 (the nut is rotatable relative to the flange) and drives the piston during tightening; this nut may be preferably bronze or steel, optionally coated with an anti-seizing material.
• the advantage of the nut 26 or 27 is that it allows to re-compress the bed at will, and this at any time; in particular, the nut 26, 27 makes it possible to compress the bed again during a separation step by chromatography.
• a set of washers or an anti-rotation plate 28 can be placed between the piston of the column 11 and the piston rod 24 in order to promote the axial displacement of the bed during the tightening, and also in order to better maintain a quasi-linear relationship between the tightening torque to exert on the clamping nut and the resulting pressure exerted on the compressed bed.
• the device according to the invention is used as follows. - Once the column 10 and the intermediate removable reservoir 9 in place, the suspension can be poured.
• the upper end of the column is closed by a flange 15 with a clamping clip 21a;
• the pressure is applied to the cylinder 14 which transmits it to the piston 11 to compress the chromatographic bed.
• the piston is moved into the column, the piston being able to slide in the column. Once the compression pressure of the bed reached, release the pressure on the cylinder 14; the piston 1 1 remains in place under the effect of friction in the column or through a mechanical locking system.
• the column is disconnected from the assembly by opening the fastener by clamp 21b.
• the threaded rod 24 and the threaded flange 25 are then placed on the column 10 and fixed by means of a clamping clip 21 d (the fasteners 21a, 21b, 21c and 21d being advantageously identical) and the pressure exerted on the bed is controlled as a function of the tightening torque.
• Column 10 is then ready for use as a chromatography column. When the pressure is removed at the piston, there may be a slight expansion of the bed. Recompression of the bed may be recommended, either before or during chromatography operations.
• the system according to the invention is rapid: after having poured the homogeneous suspension into the column and the reservoir, the duration to close the device and to put the bed in compression is of the order of one minute, which makes it possible to avoid the sedimentation of the suspension in the column, detrimental to the arrangement of the grain bed during the compression step. This is also particularly advantageous when several columns must be used (SMB, Varicol®), which reduces the preparation time of the separation process. Since the compression process is fast, it is not generally necessary to stir the suspension in the column before compression. In general, it is no longer necessary to re-fluidize the suspension in the tank.
• the device uses a unitary column, that is to say that does not need to be coupled to adjustment units (unlike, for example, the Amicon system).
• the device is flexible and can be used in "DAC” mode by keeping the column mounted on the compression module and in "non-DAC” mode according to the operating mode described with different lengths of bed. -
• the device allows to achieve in "non-DAC” performance that are close to the DAC systems, especially in terms of reproducibility. Investment and operating costs are reduced. -
• the operating procedures are easy and fast, reducing the risk of mishandling.
• the volume of the column which is necessary to contain the initial volume of the suspension is no longer linked to the chromatography column but to the removable reservoir.
• the hydraulic device associated with the base is compact and ready for use once the hydraulic fluid reservoir is filled and the air purged.
• the design is modular.
• the intermediate removable tank is placed between the base and the column and is dedicated to the departure suspension; it is then removed from the column. It can be sized according to the volume of suspension needed and be available in different lengths.
• the clamp fastening system eliminates the need for screwing at the flanges and thus eliminates bad operations and improves assembly speed.
• An anti-seizing coating on the threaded rod and the tap avoids blockages and makes it possible to control the pressure exerted as a function of the torque exerted on the threaded rod.
• the invention offers the possibility of adjusting the pressure linearly as a function of the torque exerted on the piston. - All parts of the device in contact with solvents and products to be separated are made of stainless steel or PTFE, which allows compatibility with conventional chromatographic solvents. The process is simple and fast.
• the method is a simple method for maintaining a satisfactory bed stability over time (and thus a constant column separation power).
• Column 10 is usable in all systems, especially in an SMB separation process, Simulated Moving Bed (US 2,985,589, US 3,291,726, US 3,268,605 and US 3,266,604 belonging to UOP, US 5,578,215 and US 5,578,216 belonging to IFP and NOVASEP, EP 471,082 and EP 563,388 belonging to DAICEL) or in a process by VARICOL (US 6,136,198). Such methods implement a set of these associated columns loop.
• Column 10 can also be used in batch chromatographic or CYCLOJET methods (US 6,063,284 and US 5,630,943).
• the tank is fixed at the bottom of the compression system, and then fixed below the column with a fastening system by clamp.

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• Chemical & Material Sciences (AREA)
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• General Health & Medical Sciences (AREA)
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• Treatment Of Liquids With Adsorbents In General (AREA)

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• 2006
  • 2006-03-03 FR FR0601907A patent/FR2898064A1/fr not_active Withdrawn
• 2007
  • 2007-03-02 US US12/280,914 patent/US20100163490A1/en not_active Abandoned
  • 2007-03-02 WO PCT/FR2007/000376 patent/WO2007099237A2/fr active Application Filing
  • 2007-03-02 CN CNA2007800076795A patent/CN101395468A/zh active Pending
  • 2007-03-02 EP EP07731075A patent/EP1994404A2/de not_active Withdrawn

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