PROCESS FOR CHROMATOGRAPHY ON A GEL OR ORGANIC LIQUID

专利摘要:
The invention relates to a chromatography process in which a gaseous, liquid or supercritical mobile phase containing species to be separated is circulated through a packing, said packing comprising: a plurality of capillary ducts passing through the packing between a so-called face upstream through which the mobile phase enters the lining and a so-called downstream face through which the mobile phase leaves the lining, and - a continuous medium permeable to molecular diffusion extending between said ducts, comprising a porous organic gel or an organic liquid and including at least one network of related pores whose size is greater than twice the molecular diameter of the species to be separated and open on the conduits, so as to provide said species with a diffusive path between said conduits. The invention also relates to a packing for the implementation of such a method and a method of manufacturing such a packing.
公开号:FR3026312A1
申请号:FR1459175
申请日:2014-09-29
公开日:2016-04-01
发明作者:Francois Parmentier
申请人:Francois Parmentier；
IPC主号:

专利说明:

[0001] BACKGROUND OF THE INVENTION The present invention relates to a method of chromatography on a gel or an organic liquid. BACKGROUND OF THE INVENTION Chromatography is a method of molecular separation for separating mixed species in a sample under the contradictory action of dynamic entrainment of said species by a current of a mobile phase (also called "eluent phase" ) and a retention of said species by a stationary phase. The retention conditions depend on the chemical affinity between each species and the stationary phase.
[0002] Chromatography is characterized in that it requires a great regularity of the stationary phase and its support and weak characteristic diffusional distances. This diffusional distance is usually the size of the solid grains of a particulate bed or the diameter of an empty capillary tube. In practice this distance is always less than 0.5 mm. This results in a number of theoretical separation stages greater than 200 in general. Chromatography is a particular technique, which has its own advantages and constraints and differs from other techniques that use solid packings and fluids, such as adsorption and heterogeneous catalysis.
[0003] In adsorption it is sought to retain a compound of a fluid effluent on the surface of which it is adsorbed via an isotherm, or on which it reacts. We seek to purify the fluid. We need specific high surfaces. High capacity beds are needed The effectiveness of the packing is not critical (number of theoretical trays) and it is preferred to use pellet beds of 1 to 2 mm in diameter. Effectiveness has only a negligible influence on the dimensioning of the bed insofar as it will only play on the stiffness of the percolation front, which is good as soon as one reaches about 20 theoretical plates. It is then necessary to regenerate the adsorbent by a combination of means, temperature or chemical reaction, which eliminates the adsorbed or combined impurities. The operation is therefore sequential but the cycle times are in days or weeks. We size on the mass of the bed. The pressure drops are low. In catalysis, it is sought to perform a chemical reaction on the surface of the solid. It is desired that the reagents remain an optimal time in contact with the solid. These are again adsorption forces and chemical reactions. We are interested in living time criteria. The reasoning in number of theoretical plateaux is inoperative. The regularity of the packing is one factor among others and is secondary to the catalytic selectivity. We do not try to separate molecules. The losses are low. In chromatography, several species present in a sequentially admitted fluid charge are separated by a short time interval in minutes, propagating it from an entry point to an exit point of a solid column under the effect. of an eluent fluid. The separation obtained can be achieved by a very wide variety of forces that compete with the driving effect of the eluent, sharing, adsorption, steric interactions, ionic interactions, etc. This method offers a high separating power, each component behaves differently. To emphasize this separating power, the column must have a high number of theoretical plates, for example 1000. This also means that the diffusion resistances must be minimized, and therefore that the diffusion distances are short, and that the column must be long. These combined factors make chromatography a technique which calls for an excellent regularity of the flow and therefore of the packing, and a small characteristic dimension thereof, leading to loss of charges which become rapidly critical with particulate solids. These are the problems that must be solved in chromatography. On the other hand, preparative chromatography in a practical and simple way is one of the essential problems of chemical engineering. U.S. Patent 4,957,620 to Cussler E. discloses the use of polymeric hollow fiber bundles for use as a chromatographic column.
[0004] However, the hollow fibers act independently of one another as chromatographic columns. As a result, the differences in behavior between the fibers lead to very bad efficacies. The fibers have little or no contact and do not communicate by diffusion. They are stacked in a very compact way. Molecular diffusion can not occur between the conduits. This explains the very low number of theoretical plates of the separations obtained, of the order of 40, compared to the maximum expected on a single fiber, of the order of 6000. The publication "Hollow-Fiber Liquid Chromatography" of Hongbing Ding and E. Cussler, AlChE Journal, 1989, Vol 35, No. 5, pp 814-820, details the basis of the previous patent. It is clearly stated on page 815 that the tests are conducted on 4 cm diameter modules containing 27000 hollow fibers of 100 μm internal diameter and 30 μm wall thickness. A simple calculation shows that the hollow fibers do not have a compact stack, and that the volume outside the fibers represents more than 50% of the total volume of the module. It is also explicitly mentioned on page 815, last 3 lines that the solvent constituting the stationary phase wets the hydrophobic fibers, but does not flow through the fiber towards the outside of the fiber. The volume outside the fibers thus remains filled with gas, in this case air, and empty of solvent. As a result, resistance to transfer of radial material between the fibers increases, causing a decrease in the efficiency of the separation. U.S. Patent No. 4,007,138, to Kanig G. discloses a method of making PS-DVB gels (PolyStyren, DiVinylBenzene) provided with a polymeric backing matrix. US Patent No. 8,017,015 to Clarke et al describes the state of the art processes for manufacturing organic gels and their implementation in chromatography columns. U.S. Patent No. 7,922,908 describes the use of X-rays to initiate polymerization of the organic gel. This method is particularly useful for producing solid packings. It will also be noted that the polymerization temperature may be close to ambient, from 50 ° C. to 90 ° C. Patent No. 7,473,367 to Xie S. describes processes for obtaining organic gels. Parmentier patent application VVO 2011/114017 has a packing for chromatography consisting of a monolithic porous packing. It describes in its examples a thermosetting polyester resin packing achievable according to the state of the art. However, this packing consists of a rigid and non-porous polymer that does not allow appreciable diffusivity. Indeed the publication [1] provides permeability and diffusivity measurements of thermosetting polyester. As a result, the water in this material has a diffusivity of 0.6 10-12 m 2 / s, corresponding to a permeability of 750 Barrer. This permeability is similar to that of polyethylene or polycarbonate, materials recognized as sealed and non-porous used to make containers or impermeable walls. The thermosetting polyester thus does not allow a molecular diffusion exchange of material between adjacent conduits, the effectiveness of such packing in chromatography will be limited. It is apparent from the state of the art that these organic monoliths are difficult to manufacture, particularly in large dimensions, and features that are difficult to reproduce. They are also sensitive to pressure applied and swollen in the presence of solvents or molecules to be separated. Their pore size is difficult to adjust. Their mechanical fragility makes them unable to withstand significant compressive forces. They can not therefore be used in chromatography columns operating under high pressure drops, greater than a few bars. Current packings are often particulate, have a high pressure drop, and are therefore limited in particle diameter and therefore efficiency.
[0005] Organic monoliths, however, have many advantages over silica, particularly in terms of pore size and chemical stability at high pH and because of their complete insolubility in water. There remains the need to provide stable, efficient and inexpensive organic monoliths that can be manufactured reproducibly with large dimensions. BRIEF DESCRIPTION OF THE INVENTION The invention proposes a chromatography process in which a gaseous, liquid or supercritical mobile phase containing species to be separated through a packing is circulated, said packing comprising: a plurality of through capillary ducts; the lining between a so-called upstream face through which the mobile phase enters the lining and a so-called downstream face through which the mobile phase leaves the lining, and - a continuous medium permeable to molecular diffusion extending between said ducts, comprising a gel organic porous or organic liquid and including at least one network of related pores whose size is greater than twice the molecular diameter of the species to be separated and open on the ducts, so as to provide said species a diffusive path between said ducts. Advantageously, the average molar diffusion rate of the species to be separated between the adjacent ducts under the effect of a given concentration difference of said species between the walls of said ducts is greater than 0.01 times the average molar diffusion rate of the species between a duct and the stationary phase that constitutes the lining under the effect of the same difference in concentration of the species to be separated between the fluid led by the ducts and the wall of said ducts.
[0006] Preferably, the permittivity of said continuous medium vis-à-vis the species to be separated is greater than 5000 Barrer, that is to say greater than 5 e-7 (cm3 02 cm) / (cm2 cm Hg). According to one embodiment, the diameter of the capillary ducts of the lining is less than or equal to 500 μm, preferably less than or equal to 150 μm and more preferably less than or equal to 50 μm.
[0007] According to one embodiment, said continuous medium is formed of an organic gel, said organic gel being chosen from: (a) a copolymer of styrene and divinylbenzene, (b) polymethyl methacrylate, (c) a copolymer of methacrylate hydroxyethyl and divinylbenzene.
[0008] According to another embodiment, said continuous medium is formed of an organic gel, said organic gel being a polyholoside.
[0009] According to another embodiment, said continuous medium is formed of an organic liquid extending in said network of related pores, said organic liquid being chosen from: (a) an aliphatic or aromatic hydocarbon, (b) an aliphatic alcohol or aromatic, (c) an aliphatic or aromatic ketone, (d) an aliphatic or aromatic amine, (d) a halogenated organic compound. The lining may comprise a molecular diffusion-permeable organic gel monolith through which said capillary conduits extend, said network of related pores extending within said organic gel. Alternatively, the packing comprises a monolith of a chemically inert porous material containing said network of related pores, said pores being filled with said organic gel or said molecular diffusion permeable organic liquid. Alternatively, the packing comprises a monolith of a porous chemically inert material containing said continuous pore network, the surface of said pores being covered with the organic permeable gel to molecular diffusion over a thickness selected so as to leave in said pore network a free volume for the diffusion of the mobile phase, said organic gel forming a continuous network of pores between the conduits. Preferably, the chemically inert material of said monolith is selected from silica, alumina, or a combination of silica and alumina. According to one embodiment, the lining comprises a stack of porous fibers each comprising a lumen forming a capillary duct of the lining and a wall comprising a network of related pores, said fibers being made joined by the porous organic gel or the organic liquid permeable to molecular diffusion. The wall of each fiber may be formed from said organic permeable gel. Alternatively, the pores of the wall of each fiber are filled with said gel or said organic liquid permeable to molecular diffusion. As a variant, the pore surface of the wall of each fiber is covered with the organic gel permeable to molecular diffusion over a thickness chosen so as to leave, in said pore network, a free volume for the diffusion of the mobile phase, said organic gel forming a continuous network of pores within said wall. According to one embodiment, the organic permeable gel for molecular diffusion forms the chromatographic stationary phase.
[0010] Alternatively, the organic gel has pores containing a solid third body forming the chromatographic stationary phase. Another subject of the invention relates to a method of manufacturing a packing for the implementation of the chromatography method described above, comprising the following steps: - supply of a bundle of son called precursors of the capillary ducts, - formation a porous matrix around the wires or ducts, so as to form a monolith, - removing the wires so as to form said capillary ducts.
[0011] The matrix is advantageously an organic gel. Alternatively, the matrix comprises a chemically inert material and is loaded with said matrix of an organic gel. The precursor son of the capillary ducts are advantageously fusible son at a temperature below the degradation temperature of the matrix and the elimination of said son comprises melting and draining said son out of the lining. For example, fusible wires include indium, bismuth, tin, gallium, silver or an alloy thereof with other metals other than lead, mercury and cadmium. Another subject of the invention relates to another method of manufacturing a packing for carrying out the chromatography process described above, comprising the following steps: - supply of a compact bundle of hollow fibers, - inclusion in the porous wall of the hollow fibers of an organic gel or a precursor of said organic gel to be polymerized in situ, so as to leave the lumen of the hollow fibers free and open, - creation of a diffusive bonding between said hollow fibers by said gel or organic liquid. Another object of the invention relates to another method of manufacturing a packing for carrying out the chromatography process described above, in which the organic gel is molded into a structure defining said capillary ducts. Another subject of the invention relates to a packing for chromatography, comprising: a plurality of capillary ducts passing through the packing between a so-called upstream face intended for the entry of the phase in the packing and a so-called downstream face intended for the exit of the mobile phase of the packing, and - a continuous medium permeable to molecular diffusion extending between said conduits, comprising a porous organic gel or an organic liquid and including at least one family of related pores. Advantageously, the diameter of the capillary ducts of the packing is less than or equal to 500 μm, preferably less than or equal to 150 μm and more preferably less than or equal to 80 μm. When the continuous medium is formed of an organic gel, said organic gel may be chosen from: (a) a copolymer of styrene and divinylbenzene, (b) polymethyl methacrylate, (c) a copolymer of hydroxyethyl methacrylate and divinylbenzene. Alternatively, the organic gel may be a polysaccharide. When said continuous medium is formed of an organic liquid extending in the network of related pores, said organic liquid is selected from: (a) an aliphatic or aromatic hydocarbon, (b) an aliphatic or aromatic alcohol, (c) a aliphatic or aromatic ketone, (d) an aliphatic or aromatic amine, (d) a halogenated organic compound.
[0012] According to one embodiment, the lining comprises a molecular diffusion permeable organic gel monolith through which said capillary ducts extend. According to another embodiment, the lining comprises a monolith of a porous chemically inert material having a continuous network of pores, said pores being filled with said gel or said organic liquid permeable to molecular diffusion. According to another embodiment, the lining comprises a monolith of a porous chemically inert material having a continuous network of pores, the surface of said pores being covered with the organic gel permeable to molecular diffusion to a thickness chosen so as to preserve, in said pore network, a free volume for the diffusion of the mobile phase, said organic gel forming a continuous network of pores between the conduits. According to another embodiment, the lining comprises a stack of porous fibers each comprising a lumen forming a capillary duct of the lining and a wall comprising a continuous network of pores, said fibers being made joined by the gel or organic liquid permeable to diffusion. molecular. The wall of each fiber can then be formed of said organic permeable gel molecular diffusion.
[0013] Alternatively, the pores of the wall of each fiber are filled with said gel or said organic liquid permeable to molecular diffusion. BRIEF DESCRIPTION OF THE DRAWINGS Other characteristics and advantages of the invention will emerge from the detailed description which follows, with reference to the appended drawings, in which: FIG. 1 is a sectional view of a schematic diagram of a lining; monolithic according to one embodiment of the invention in a plane parallel to the longitudinal axis of said lining, - Figure 2 is a sectional view of a schematic diagram of said lining in a plane perpendicular to the longitudinal axis of said lining FIG. 3 is a sectional view of a schematic diagram of a multicapillary packing comprising a dimensionally stable porous skeleton whose pores are covered with a molecular diffusion-permeable organic gel; FIG. sectional diagram of a packing formed of a stack of hollow fibers according to one embodiment of the invention in a plane perpendicular to the longitudinal axis of said gar 5 and 6 are top and sectional views of a molded organic gel; FIG. 7 shows the effectiveness of a multicapillary lining in which the wall of the ducts is non-porous (a) and porous; (b). FIGS. 8 and 9 show the diffusive flows between adjacent ducts and inside the same duct.
[0014] DETAILED DESCRIPTION OF THE INVENTION The chromatography method uses a stationary phase which is in the form of a packing of which different embodiments will be described below and a mobile phase containing species to be separated. Chromatography is a particular molecular separation process, characterized in that it separates a mixture of chemical substances under the contradictory action of a dynamic entrainment of these species by a current of an eluting phase. retention of these species by a stationary phase.
[0015] Preferably this process is continued until complete elution of the separated species out of the stationary phase.
[0016] Advantageously, a chromatographic separation or a chromatographic method will advantageously be characterized in that it comprises at least 300 theoretical stages, and preferably at least 1000 theoretical stages. This distinguishes it from membrane separations, catalytic processes and adsorption or ion exchange separations in particular. The number of optimal theoretical stages NET of a process for a non-retained compound can advantageously be calculated by the formula: ## EQU1 ## In which L is the length of the column, Dh is the average hydraulic diameter (arithmetic mean) of the ducts, e is the average wall thickness, P is the void fraction of the walls. Chromatography is applied in liquid, gas and supercritical phases. The invention makes it possible to optimize the efficiency and speed of a chromatographic process by selecting the morphology and the structure of the most advantageous packing. The invention relates to a chromatography method in which a mobile phase containing species to be separated is circulated through a lining, said lining comprising: a plurality of capillary ducts passing through the lining between a so-called upstream face through which the phase mobile penetrates into the lining and a so-called downstream face through which the mobile phase leaves the lining, and - a continuous and connected medium permeable to molecular diffusion extending between said ducts, comprising a porous organic gel or an organic liquid and comprising a pore family providing connectivity between said conduits.
[0017] The process therefore comprises an exchange of material between the mobile phase and a stationary phase, which may consist of the gel or organic liquid itself or of a solid third substance contained in the porosity of the organic gel. By "permeable to molecular diffusion" is meant that the conduits of the lining are connected by a continuous and connected phase comprising at least one porous organic gel and / or an organic liquid on the one hand and optionally a mobile phase on the other hand , and having a family of pores providing connectivity between said conduits. Advantageously, the pore size of the continuous medium is greater than twice the molecular diameter of the species to be separated.
[0018] Advantageously, the pore size of the continuous medium is greater than 10 times the molecular diameter of the species to be separated. Advantageously, the pore size of the continuous medium is greater than 2 times and less than 1000 times the molecular diameter of the species to be separated.
[0019] Advantageously, the pore size of the continuous medium is greater than 2 times and less than 30 times the molecular diameter of the species to be separated. Advantageously, the continuous and connected phase linking the conduits is a condensed phase.
[0020] Advantageously, the continuous and connected porous medium extending between the walls does not present any material interruption. The porosity of the material can be advantageously defined by chromatography in three ways: 1. The porosity of an organic gel can come from the swelling of a crosslinked gel in an organic solvent, mineral or aqueous, swelling advantageously representing more than 2% of its volume, and preferably more than 10% of its volume. 2. It may come from a porosity of the gel in the unsolvated state. 3. It can come from the porosity of a support on which a polymeric gel is deposited in the form of a thin layer. Within this definition of the molecular permeability, it is advantageously understood that under the conditions of the chromatographic process and for the species to be separated: on the one hand, the average molten flow rate Phip between the adjacent ducts under the effect of a given concentration difference of the species to be separated between the walls of said ducts, and on the other hand the average molar flow rate; Phic diffusion between a conduit and the stationary phase that is the packing under the effect of the same difference in concentration of the species to be separated between the fluid conducted by the conduits and the wall of said conduits. are close to each other, mean flow means that the flow is the average of this flow measured on the lining as a whole. By convention, it will be considered that the Phic diffusion molar flow rate between a pipe and the stationary phase constituted by the packing is measured by imposing a uniform concentration on the wall Cs and by calculating the exchange with respect to the average concentration Ce fluid flowing through the duct. This results in a Sherwood number equal to 3.66 in the case of a circular section tube traversed by a fluid in laminar flow. By close to each other is meant in particular that said molar mass transfer rate of the species to be separated between two adjacent ducts between their walls is at least 0.01 times, advantageously at least 0.1 times and again more preferably at least 0.5 times the term of said molar flow rate related to the transfer of material between the ducts and their stationary phase.
[0021] In the present text, the molecular diameter will be calculated in two ways depending on the molecular weight of the substance under consideration. For substances with a gaseous phase and whose critical coordinates are accessible, we will use the covolume, term b of the Van der Vals equation, divided by 4 and by the number of Avogadro, and we will calculate the diameter of a sphere of equivalent volume. It is known that the covolume b is equal to four times the molecular volume. The covolume is easily accessible from the critical coordinates of the body. For macromolecules, biological molecules (proteins, etc ...) and molecules whose critical coordinates are inaccessible, we will use the hydrodynamic diameter measured by dynamic light scattering. The mobile phase is, under the conditions of implementation of the process, in the gaseous state, liquid or in the supercritical state. Preferably, the mobile phase is, under the conditions of implementation of the method, in the liquid state or in the supercritical state. More preferably, the mobile phase is, under the conditions of implementation of the method, in the liquid state. Indeed, the effectiveness of a lining increases proportionally to the density and diffusivity of the mobile phase that passes through it. In order to increase the effectiveness of the packing, the density of the mobile phase passing through it is increased by working in the vicinity of the critical point of said mobile phase or in the liquid state. Furthermore, the lining has sufficient strength, rigidity and mechanical strength to allow handling of the lining. The packing may advantageously be implemented on an industrial scale under pressure losses of between one and a few bars per meter. The packing used in the present invention is a porous lining which comprises a plurality of parallel capillary ducts which extend in the direction of circulation of the mobile phase, this direction being considered as the longitudinal direction of the lining. Such packing is called "multicapillary".
[0022] In such a lining, the capillary ducts are advantageously empty of solid material while the material which surrounds the ducts is porous. In particular, at least the wall of the ducts has a continuous network of pores, said pores being open on the ducts. The capillary ducts are advantageously rectilinear, although it is not excluded that the ducts have bends or angles. The capillary ducts have a uniform section with respect to each other and their length.
[0023] The section variability of the ducts will be conveniently defined by a relative standard deviation. This relative standard deviation represents the ratio of the standard deviation of the pipe diameter to the average pipe diameter, expressed as a percentage. Advantageously, the ducts have a substantially constant mean diameter from one duct to the other, such that the standard deviation of the diameter on the duct duct sample does not exceed 30% of the average diameter, preferably does not exceed 10%. the average diameter, and even more preferably does not exceed 2.0% of the average diameter. In the present text, the mean of a set of values of a variable X is its arithmetic mean E [X]. The standard deviation is defined as the square root of the arithmetic mean of (X-E [X]) 2. By distribution, is meant in the present text a set of values of the variable X. Advantageously, the diameter does not vary by more than 50% over the length of the same conduit. Advantageously, the diameter preferably does not vary by more than 20% over the length of the same conduit. Even more advantageously, the diameter does not vary by more than 10% over the length of the same conduit. More preferably, the diameter does not vary by more than 2% over the length of the same conduit. Advantageously, the ducts pass through the packing from one end to the other, thus making it possible to minimize the pressure drop within the packing during the chromatographic separation process. Advantageously, the volume of the capillary ducts represents more than 5% of the total volume of the lining, preferably more than 30% of said total volume and more preferably more than 50% of the total volume of the lining. The term "total volume of packing" in the present text means the volume occupied by the packing, including its porosity; said total volume can therefore be calculated from the outside dimensions of said lining. The volume of the ducts is measured in the following manner: number of ducts x average section of a duct x average length of a duct. Advantageously, the volume occupied by the organic gel in the packing is greater than 2% of the volume of the packing excluding the ducts, preferably greater than 10% of said volume and more preferably greater than 40% of said volume. . By "packing volume excluding ducts" is meant the difference between the total volume of the lining and the volume of the capillary ducts. The conduits may have a section of any suitable shape, for example a circular, square, rectangular, hexagonal, star, slot-shaped, etc. When the ducts have a non-circular section, the term "diameter" of said ducts their hydraulic diameter. Advantageously, the ducts have a hydraulic diameter of less than or equal to 500 μm. According to one embodiment, the hydraulic diameter of the ducts is less than or equal to 150 μm, or even less than or equal to 50 μm. The hydraulic diameter is conventionally calculated as equal to four times the section of a duct (in m2) divided by the perimeter of said duct wetted by the mobile phase (in m). For a liquid chromatography process, the conduits preferably have a diameter of less than 30 μm, and more preferably less than 15 μm. For a supercritical phase chromatography process, the conduits preferably have a diameter of less than 80 μm, and even more preferably less than 30 μm.
[0024] For a gas chromatography process, the conduits preferably have a diameter of less than 250 μm, and even more preferably less than 50 μm. In fact, the chromatography is carried out simply in gravity apparatus, or the weight of the fluid column on the packing causes its flow. The upper limit of the diameter of the capillaries will be obtained when the flow of the fluid at the speed allowing the optimum of the effectiveness of the packing will cause a loss of load equal to the weight of the column of fluid considered on the height of the bed. It is known that for a multicapillary packing with optimum efficiency: 20 Vdc TT vR Do The Poiseuille law is written Ap = 32, g, LG, vc 25 The pressure caused by a fluid height LG s' Ad) = p * g * LG It follows that: 30 dm '= 3, / 32, g, D0WR P * 9 The table below exemplifies dmax for various fluids common in chromatography. Solvent II (P Do m2 / s) VR p (kg / m3) dmax p Water 0.001 1E-09 5 1000 25.3869139 Hexane 0.00031 3E-09 5 659 28.4754613 Methanol 0.00055 2E-09 5 791 28 , 3363475 Chloroform 0.00057 2E-09 5 1480 23,2716977 VR is in general between 2 and 5. It will be possible to take a simplified way the value of 50 pm as cut-off threshold of the diameter of the ducts making it possible to take advantage advantageously of the advantages of multicapillary chromatography. In the same simplified way, it will be possible to take the value of 80 μm as the cut-off point of the inside diameter of the ducts taking into account the thickness of the stationary phase. In these formulas V, is the mobile phase velocity in the duct, d, is the mean internal diameter of the duct, C is the diffusivity of the species to be separated in the mobile phase, the is the viscosity of the mobile phase, p is the density of the mobile phase, LG is the length of the column, g is the acceleration of gravity, AP is the pressure drop of the fluid in the conduit, dm 'is the maximum permissible mean internal diameter for a chromatographic separation. All these quantities are expressed in the SI unit system. Advantageously, the conduits are distributed in a single mode, that is to say around a single average diameter. Without departing from the scope of the invention, the diameter of the ducts can however be distributed over several modes, and in particular over two modes. The known laws connecting means and variances of several populations can be applied in order to respect the previous numerical constraints connecting mean and relative standard deviation.
[0025] Moreover, the ducts may be arranged regularly in a regular square or triangular axial mesh. By this is meant that the axes of the conduits are arranged at the vertices of squares closely stacked with substantially constant sides or equilateral triangles closely stacked with substantially constant sides.
[0026] Finally, the relative standard deviation of the thickness of the wall separating two adjacent ducts measured on a section of the lining is preferably less than 30%, even more preferably less than 10%, and even more preferably less than 2.0%. . In this case, the relative standard deviation characterizes the ratio between the standard deviation of the thickness of the wall and its average, expressed in%.
[0027] Advantageously, the lining may have any specific surface area of between 0.1 and 1200 m 2 / g. Advantageously, when the packing comprises a porous monolithic organic gel acting as a stationary phase by adsorption on its porous surface, this surface will preferably be greater than 60 m 2 / g, and even more preferably between 80 and 600 m 2 / g. This surface may be rough or modified by surface chemical treatment. Advantageously, when the lining comprises an organic gel supported by an underlying structure and acting in its mass by penetration of the molecules to be separated in its volume, the support structure of the organic phase will have a specific surface area preferably of less than 60 m 2 / g preferably less than 20 m 2 / g, and even more preferably less than 2 m 2 / g. Advantageously, the average pore size of the walls of the lining ducts will be between a few Angstroms and several hundred nanometers depending on the needs and the type of chromatography used. The pore volume of the walls of the lining will advantageously be between 0 cm 3 / g (this is the case when the walls of the lining are filled with a liquid stationary phase for example) and several cm 3 / g (case of PS-type polymeric stationary phases). As mentioned above, the invention uses a packing which comprises a gel or an organic liquid permeable to molecular diffusion. This gel or organic liquid may be the constituent material of the packing, or may form only a part of the packing, for example as impregnation of the porosity of a porous material of different nature or as a coating layer deposited on a porous material of different nature so as to cover the pores of said material. According to one embodiment, the lining is a monolith formed of said organic gel. In this packing, the organic gel forms a continuous skeleton defining a continuous network of pores extending between the ducts and open on the ducts. According to another embodiment, the lining is a monolith comprising a porous skeleton formed of a material other than an organic gel and whose pores are filled with a gel or organic liquid. Thus, the gel or organic liquid forms a continuous medium permeable to molecular diffusion extending between the conduits.
[0028] Alternatively, the packing is a monolith comprising a porous skeleton formed of a material other than an organic gel and whose pore surface is covered with an organic gel film, such that the pores of the material contain a residual volume devoid of organic gel, free for the diffusion of the mobile phase. According to another embodiment, the lining comprises a stack of porous and hollow fibers. Each fiber comprises a lumen forming a capillary duct and a wall comprising a continuous network of pores. Advantageously, these fibers are stacked compactly so as to occupy more than 60% of the total volume of the lining, and preferably more than 80% of the total volume of the lining.
[0029] According to one embodiment, each fiber consists of an organic gel. Alternatively, each fiber is coated or impregnated with an organic gel.
[0030] Preferably, said fibers are joined by means of the gel or organic liquid permeable to molecular diffusion. Compared to the packings consisting of a porous mass of organic gel devoid of capillary ducts, a multicapillary packing of organic gel has, at equal efficiency, a significantly lower pressure drop (of the order of 10 to 30 times). Consequently, the formation of capillary ducts in the packing makes it possible to compensate for the relatively low mechanical strength of the organic gel packing by reducing the pressure drop and consequently obtaining moderate mechanical stresses exerted on the packing during the filling. chromatography process. To ensure good mechanical cohesion of the lining, the capillary ducts are advantageously mechanically secured by means of a continuous solid medium between the ducts and containing a gel or organic liquid permeable to molecular diffusion. This continuous medium creates a bond between the conduits that allows the molecular diffusion to proceed freely between adjacent conduits. As will be explained below, the implementation of a diffusive exchange between the ducts makes it possible to increase the number of theoretical plates available for a chromatographic separation process, and thus to increase the efficiency of such a method. . The implementation of a diffusive exchange between the conduits makes it possible to level the differences of behaviors between individual conduits. It is therefore necessary to increase the intensity of these exchanges in order to obtain an increase in the efficiency measured in terms of theoretical platforms available. The increase in this efficiency results in a greater separating power of the lining with respect to a given mixture. A mixture of closely related species of species can be solved more easily, and these species separated with a greater degree of purity. Since the organic gel is subject to some swelling between the dry state and the wet state, it is advantageous, in order to avoid variations in the diameter of the conduits, to strengthen the organic gel by means of a dimensionally structured structure. more stable than said organic gel. By "dimensionally stable structure" is meant a structure with no or little mechanical deformation in the conditions of use of the packing. In particular, this structure does not exhibit a significant swelling effect in the presence of the eluent solvents characteristic of a chromatographic process in the liquid or supercritical phase, said swelling remaining advantageously less than 10% of the total volume of the lining, and preferably less than 2% of said total volume.
[0031] A dimensionally stable structure that can be used to obtain a packing adapted to the implementation of the invention comprises the highest possible pore volume intended to serve as a reservoir for the gel or organic liquid. Said pore volume is preferably greater than 20% of the volume of the packing excluding ducts, preferably greater than 40% of this volume, and still more preferably greater than 60% of this volume. Advantageously, such a dimensionally stable structure has a chemically inert surface and a low specific surface so as not to interfere with the chromatographic process taking place in the organic gel. Advantageously, this specific surface area will be less than 20 m 2 / g, preferably less than 2 m 2 / g, and even more preferably less than 0.2 m 2 / g. According to one embodiment, the dimensionally stable structure is a fabric or a nonwoven made of a mineral or organic material. Advantageously, this fabric is made of one or more structural fibers such as fiberglass, carbon fiber, aramid fiber, metal fiber or a mixture thereof. To form said structure, a weaving technique is advantageously used: precursor fibers of the ducts (that is to say having an outside diameter equal to the diameter of the ducts and intended to be subsequently destroyed to form the ducts) are woven in wefts and structural fibers are arranged in a chain, or vice versa. The fabric is shaped, for example rolled up or stacked. It is impregnated with precursor liquid of the organic gel or a porous matrix of another material. The precursor liquid is polymerized or the porous matrix is bonded to give the assembly optimum mechanical strength. The structural fiber supports the stresses associated with the swelling of the material and improves its longevity and dimensional stability in operation. Alternatively, the dimensionally stable structure is a fiber bundle of inorganic or organic material. According to another embodiment, the dimensionally stable structure is a reinforcing filler such as fibers, microfibers, cut or milled nanofibers, precipitated silicas, a diatomaceous earth, etc. Among the cut fibers, chopped or crushed, note the polyolefin fibers, glass, silica, aramids, metal. According to one embodiment, the dimensionally stable structure constitutes the rigid skeleton of the lining. This dimensionally stable structure may be a porous monolithic multicapillary structure such as a ceramic, metal or polymer monolith. Among the possible ceramics, in particular, but not limited to, titanium oxide, zirconium oxide, alumina, aluminosilicates, cordierite, mulite, silica, glass, metal silicates such as zinc silicates , magnesium, calcium, aluminum, titanium, zirconium, etc. or the aluminosilicates of these metals. These monoliths can be obtained by known methods, for example by extrusion and sintering. However, these extrusion processes are poorly suited to producing ducts of diameter less than 0.8 to 1.2 mm and are therefore not very effective with respect to diffusive phenomena. In particular, they will be unproductive. Advantageously, said monoliths are obtained by any of the processes described in applications VVO 2011/114017 and VVO 2013/064754 of Parmentier. Such monoliths advantageously comprise the highest possible pore volume in order to serve as a reservoir for the gel or organic liquid, preferably greater than 20% of the volume of the packing excluding the ducts, advantageously greater than 40% of this volume, and more preferably still greater than 60% of this volume. The multicapillary monoliths for carrying out the invention will advantageously be based on networks of organic or inorganic polymers.
[0032] The multicapillary monoliths for carrying out the invention will advantageously be based on networks of highly crosslinked polymers or on networks of silica or porous alumina or one of their combinations. These networks can be made in the same way as the currently called monolithic packings for chromatography, which are macroporous non-multicapillary high porosity monoliths. By combination of silica and alumina is meant any formulation containing mainly silica and alumina in combined form, or in the form of a mixture, or simultaneously in combined form and in the form of a mixture. In particular, the multicapillary monolith may consist of a bimodal silica. This bimodal silica consists of a three-dimensional mesoporous porous skeleton in which a volume of interconnected macropores is inscribed. In this case, the organic gel is possibly occluded in the volume of the mesopores. By "mesopores" are meant pores whose diameter is between 2 and 50 nanometers; "Macropores" means pores whose diameter is greater than 50 nanometers; "Micropores" means pores whose diameter is less than 2 nanometers. The pore sizes mentioned in the present text are measured according to two different techniques depending on the nature of the material tested: when it is a mineral material and in particular silica, the technique used is mercury porosimetry for the macro and mesoporosity, and nitrogen adsorption for microporosity; in the case of polymeric materials or based on inorganic matrices coated with organic gels, mercury porosimetry is used for macroporosity and nitrogen adsorption porosimetry for mesoporosity and microporosity.
[0033] According to one embodiment, the organic gel covers the mesopore skeleton. The macropore volume advantageously remains open and interconnected so as to ensure free diffusion of the species contained in the mobile phase in the mass of the monolith.
[0034] Advantageously, the volume of the mesopores and macropores is increased. It is also possible to stabilize a silica support monolith by precipitation or adsorption of zirconium oxide on its surface so as to render it stable at alkaline pH. Advantageously, bi-modal silicas are used as support for the organic gel or the organic liquid, the mesopore volume of which is between 10% and 40% of the total pore volume. Advantageously, bi-modal silicas are used as support for the organic gel or the organic liquid, the macropore volume of which is between 60% and 90% of the total pore volume. When the organic gel is included in a porous structure, it may consist of molecules of low molecular weight, advantageously less than 1000 g / mol, more advantageously less than 500 g / mol, and even more preferentially less than 150 g / mol . These low molecular weight substances may in this particular case be organic liquids. In the case of liquid chromatography, the organic liquids that can be used as organic gel for carrying out the invention will be in particular aldehydes, ketones (methyl ethyl ketone, methyl isobutyl ketone, methyl cyclohexanone, dimethyl cyclohexanone). ), esters (cyclohexyl acetate, furfuryl acetate, amyl acetate), ethers (2-chloro-2-methoxy diethyl ether, diisopropyl ether) aliphatic and aromatic hydrocarbons (hexane, dodecane, and benzene, toluene) alcohols (iso-butanol) pentanol, octanol, dodecanol, methyl cyclohexanol, 2-ethyl hexanol), carboxylic acids (octanoic acid, naphthenic acids). Other organic liquids can be used, such as tributyl phospahate, trioctyl phosphate, trioctyl phosphine oxide, phosphonic acid esters, dimethyl phthalate, diethyl oxalate, aryl sulfonic acids, hydroxyoximes, oximes derivatives, beta-diketones, alkylaryl sulfonamides, primary, secondary, tertiary, quaternary amines, etc. An advantage of organic liquids over organic gels is their very homogeneous aging, which makes it possible to maintain a chromatographic process. homogeneous within the packing even when it is subject to aging. Such aging is due in particular to the presence of contaminating particles. Unlike organic gel, the organic liquid allows the diffusion of such particles in its mass so as to obtain a regular distribution within the liquid.
[0035] When the process is used to separate biological molecules, for example proteins, the organic liquid preferably contains nanometric aqueous solution micelles stabilized in the less polar phase by surfactants, referred to as inverted micelles. These reverse micelles have some pH-dependent electrostatic charge that can solubilize an oppositely charged protein. In the context of a chromatographic process, when the products to be separated are proteins and the immobilized organic phase contains such reverse micelles, the pH of the mobile phase is adjusted so that these reverse micelles solubilize a particular protein. After the substantially similar electrostatic charge proteins have been separated (as not being soluble in the reverse micelles), the protein solubilized in the reverse micelles can be eluted by changing the pH of the mobile phase. In the case of gas phase chromatography, the organic liquids that can be used to implement the invention will in particular be polysiloxanes, including methyl, benzyl, trifluoropropyl, cyanopropyl, etc., polyethylene glycols, and the like. The organic gel is sufficiently permeable to allow a high diffusivity of the species of the mobile phase between the different adjacent ducts. The efficiency of a chromatographic process is thus considerably increased by leveling the differences in the behavior of slightly different capillary ducts in diameter, in wall thickness, etc. thanks to the molecular diffusion between neighboring ducts. According to one embodiment, a continuous network of related pores is created for this purpose in the mass of the organic gel or around the organic gel. These related pores consist of mesopores, mesopores and macropores, or macropores.
[0036] Advantageously, these pores belong to at least two independent networks of mesopores and macropores. The pore volume of the packing containing the organic gel is preferably made to be greater than 20%, preferably greater than 40% and even more preferably greater than 60% of the total volume of the packing, excluding the volume of the pipes. Advantageously, although not exclusively, the organic gel is made to contain macropores larger than 50 nanometers in size. These macropores allow the molecules of the solvent and some of those of the charge to diffuse rapidly into the organic gel.
[0037] According to a particularly advantageous embodiment of the invention, the walls of the lining comprise an empty volume fraction of gel or organic liquid and, where appropriate, of stationary phase. This fraction makes it possible to serve as volume reserves with respect to swelling phenomena experienced by the organic gel in contact with a solvent or solutes by releasing the induced mechanical stresses and ensuring the integrity and the dimensional permanence of the diameter of the conduits. Indeed, the phenomena of swelling of the organic gel are absorbed and compensated macroscopically by this empty volume in the internal volume of the walls of the ducts and have no effect on the volume offered to the flow of the mobile phase. Advantageously, this fraction of empty volume is accessible to the stationary phase. Advantageously, this fraction of empty volume represents more than 5% of the volume of the organic gel, preferably more than 10% of the volume of the organic gel, and even more preferably more than 25% of the volume of the organic gel. This phenomenon is particularly advantageously used in combination with a strengthening of the organic gel by a dimensionally stable structure. In fact, the dimensionally stable structure ensures the constancy of the geometrical dimensions of the lining and in particular the diameter of the ducts, by supporting the mechanical stresses associated with the phenomena of swelling or shrinkage of the organic gel during its cycles of use. These shrinkage and swelling phenomena are thus entirely rejected towards the pore volume of the organic gel. The increase or decrease in the volume of the organic gel due to differences in solvation has an integral effect on a decrease or an increase in its pore volume while keeping constant the dimensions of its macroscopic envelope and in particular the diameters of the conduits. For this purpose, the organic gel can be deposited in the form of a thin layer on a porous support. This deposition can be done by any technique known to those skilled in the art as by soaking the dimensionally stable structure in a solution of the organic gel in a solvent, or in a solution of a precursor of the organic gel. The drainage of the structure and the evaporation of the solvent leave a thin layer of organic gel or precursor of the organic gel which can be polymerized or crosslinked in situ throughout the entire inner wall or specific surface of this structure.
[0038] This thin layer of organic gel leaves free an empty porous volume of organic gel. Advantageously, this free pore volume is continuous and connected, that is to say that its pores communicate with each other. This porous volume accessible to the mobile phase allows efficient diffusion of the dissolved molecules between adjacent ducts.
[0039] According to one embodiment, the organic gel is deposited in the form of a thin layer on the backbone of a bimodal silica structure or an organic monolith. Said thin layer is of micrometric or submicron thickness.
[0040] Said thin layer may be deposited in sufficient quantity to cover the skeleton of the monolith, but in insufficient quantity to fill the volume of the macropores. In particular, it is deposited so as to maintain a volume of macropores connected and continuous in the lining and between the ducts. Such a deposition can be carried out for example by filling the macropores and mesopores with the precursors of the organic gel in the dissolved state in a volatile solvent and evaporating the solvent uniformly before crosslinking the organic gel in situ if necessary. The deposit can take place in the whole mass of the structure or on only part of it. In particular it may consist of an organic gel layer deposited on its surface and on the wall of the ducts. In order to give the polymeric organic gel the required porosity, the polymerization can be conducted in the presence of a pore-forming agent subsequently removed. This blowing agent can be a solvent or a body subsequently removed by dissolution, evaporation or etching such as a macromolecule or a silica gel or alumina. When it is a solvent it will lead to the precipitation of the organic gel during its polymerization. When it is a macromolecule it may in particular be used to aggregate or coacerver between them particles of polymeric organic gel. The porogen may in particular be an organic solvent or water. For example, organic solvents such as alcohols, esters, ethers, aliphatic and aromatic hydrocarbons, ketones, di, tri tetraethylene glycols, butane diols, glycerols and the like can be used. Among the porogenic heavy solvents include tetrahydronahthalene, decahydronaphthalene, anthracene, biphenyl, paraffin oils, stearic acid, oleic, palmitic, dialkyl phthalates, camphor and its esters, dodecanol -1, octanol-1, cyclohexanol or a mixture thereof.
[0041] The amount of pore-forming agent may vary between 10 and 90% and preferably between 20 and 60% by volume of the final mixture comprising the monomers. The blowing agent influences the final distribution in terms of pore size. According to one embodiment, the dimensionally stable structure is bonded to the polymeric organic gel by chemical bonds, especially covalent bonds.
[0042] Advantageously, these covalent bonds are made by grafting onto the surface of the dimensionally stable structure of a coupling agent capable of reacting with the organic gel before, after or during its crosslinking or polymerization. The coupling agent may comprise vinyl, acrylic or methacrylic linkages for coupling to vinyl, styrenic or acrylic gels. Advantageously, this coupling agent comprises alcohol groups, and more advantageously oses or holoside molecules for coupling to polyholosides such as dextran or agarose during their crosslinking. Among the coupling agents which may be used, inter alia, but not limited to, are Dodecyltrimethoxysilane, octadecyltrimethoxysilane, methyltrimethoxysilane, n-octyltriethoxysilane, n-octyltrimethoxysilane, n-propyltrimethoxysilane, n-propyltriethoxysilane, methyltriacetoxysilane, ethyltriacetoxysilane, vinyltriacetoxysilane and vinyltri (2-methoxyethoxy) silane. , 3-Chloropropyltriethoxsilane, 3-chloropropyltrimethoxysilane, 35 chloropropylmethyldimethoxysilane, 3-aminopropyltriethoxysilane, 2-am Aminoethy1-3- inopropyltrimethoxysilane, 3-Aminopropyltrimethoxysilane, Bis (trimethoxysilypropyl) amine, 3-U reidopropyltrimethoxysilane, 3- glycidoxypropyltrimethoxysilane, 3-glycidoxypropylmethyldimethoxysilane, 3- Methacryloxypropyltrimethoxysilane, 3-methacryloxypropylmethyldimethoxysilane, bis (3-triethoxysilylpropyl) tetrasulfide, bis (3-triethoxysilylpropyl) disulfide, 3-mercaptopropyltrimethoxysilane, 3-mercaptopropylmethyldimethoxysilane, vinyltris (methylethylketoxime) silane, Inyl Oximino Silane, Methyltris (methylethylketoxime) silane, Methyl Oximino Silane, Tetra (methylethylketoxime) silane, Trifluoropropylmethyldimethoxylsilane, silanes containing epoxy bonds, etc. As previously mentioned, the term "permeable to molecular diffusion" is understood to mean that the conduits of the lining are linked by a continuous and connected phase comprising at least one porous organic gel and / or an organic liquid on the one hand and possibly a mobile phase on the other hand, and comprising a family of pores 20 ensuring a connectivity between said conduits Advantageously the pore size of the porous medium is greater than 2 times the molecular diameter of the species to be separated. Advantageously, the continuous and connected phase linking the conduits is a condensed phase. Advantageously, the continuous and connected porous medium extending between the walls has no material interruption. The porosity of the material can be advantageously defined by chromatography in three ways: 1. The porosity of an organic gel can come from the swelling of a gel crosslinked in an organic solvent, mineral or aqueous, swelling advantageously representing more than 2% of its volume, and preferably more than 10% of its volume. 2. It may come from a porosity of the gel in the unsolvated state. 3. It can come from the porosity of a support on which a polymeric gel is deposited in the form of a thin layer. Within this definition of molecular permeability is advantageously meant that under the conditions of the chromatographic process and for the species to be separated: on the one hand, the average diffusion flow rate Phip between the adjacent ducts under the effect of a difference in concentration of the species to be separated between the walls of said ducts - on the other hand the molar flow rate Phic average diffusion between a duct and the stationary phase that constitutes the lining under the effect of the same difference in concentration of species to be separated between the fluid driven by the conduits and the wall of said conduits. are close to each other. By convention, it will be considered that the Phic diffusion molar flow rate between a pipe and the stationary phase constituted by the packing is measured by imposing a uniform concentration on the wall Cs and by calculating the exchange with respect to the average concentration Ce fluid flowing through the duct. This results in a Sherwood number equal to 3.66 in the case of a circular section tube traversed by a fluid in laminar flow.
[0043] By close to each other is meant in particular that said molar mass transfer rate of the species to be separated between two adjacent ducts between their walls is at least 0.01 times, advantageously at least 0.1 times and again more preferably at least 0.5 times the term of said molar flow rate related to the transfer of material between the ducts and their stationary phase.
[0044] By mean flow means that said flow rates are calculated for a mean pipe diameter and a mean wall thickness of the lining, said means being arithmetic means. This amounts to expressing in other words that the term conductance related to the transfer of material between two adjacent ducts at their wall is at least 0.01 times, advantageously at least 0.1 times and even more preferably at least 0 5 times the term of conductance related to the transfer of matter between the ducts and their stationary phase Recall that Sh = k * D / Diff k is the material transfer coefficient, D the diameter of the duct, Diff the diffusion coefficient. The molar transfer rate of diffusion material Phi per unit area of the wall of the duct is deduced: Phic = k * (Cs-Ce) Advantageously, this value is measured on the components of the mixture to be separated under the conditions of separation or of the chromatographic process. In order to know the permittivity or effective diffusivity of a real wall structure, and to calculate a molar diffusion rate Phip in the wall separating two adjacent ducts, and the diffusion rate between a duct and its wall containing the stationary phase Phic, it will preferably use a computer simulation comprising all the morphological, geometric and constituent, physical and physico-chemical details of said wall and lining. Software such as COMSOL multiphysics makes it easy to achieve such performance. The input data of such a simulation are essentially - the porous fractions filled by mobile phases in the wall, the tortuosity and the average pore size and pore size distribution of these porous fractions as well as the molecular diffusivity of the species. to be separated measured in these phases under the conditions of the chromatographic separation. When these are not available experimentally, we can estimate them by the method of VVilke and Chang. The porous fractions filled with organic gel or a liquid organic stationary phase in the wall, as well as the molecular diffusivity of the species to be separated, measured in these gels if appropriate under the conditions of the chromatographic separation. - The geometry of the wall including details such as the position and the dimensions of the zones filled by the organic gel, the organic liquid and the mobile phase and any dead zones or filled by fluids or substrates other than the mobile phases, organic liquid and organic gel, as well as the molecular diffusivity of the species to be separated measured therein under the conditions of the chromatographic separation. - The partition coefficients of the species to be separated between the different phases present in the concentration range encountered during the chromatographic process. - The pressure drop applied to the packing and the composition of the eluent fluid and its viscosity under the conditions of the chromatographic separation. In a more approximate way in preliminary to a simulation one will be able to approach diffusional flows by the following equations.
[0045] The following numerical quantities can be defined: Gpar01 = K * DeffieP Gmobile = 3. 66 * Do / (D h) with K = Cstat / Cmobile and we deduce Phic / Phip> Gmobile / Gparoi where K is the partition coefficient of the considered species between the stationary phase considered to be defined by the entire volume of the wall, and the mobile phase, Cstat is the concentration of the species to be separated in the stationary phase (mole / m3), Cmobile is the concentration of the species to be separated in the mobile phase (mole / m3) at the thermodynamic equilibrium with the previous one, Deff (m2 / s) is the effective diffusivity in the material of the wall, ep (m) is the average thickness of the wall separating two ducts, C is the diffusion coefficient of the considered species in the free mobile phase, and Dh is the average hydraulic diameter of the capillary duct.
[0046] Similarly, as a preliminary to a simulation, it will advantageously be to set a condition on the ratio between the maximum number of theoretical trays of a NETMax separation and the theoretical number of trays actually observed at the optimum of NET efficiency for an unsuccessful compound. on the actual material. NETMax at optimum efficiency can be taken as equal to the formula previously given in this text. NETMax = 1.6 * L / (Dh + e * P) Advantageously, NETMax is obtained by a computer simulation. Advantageously, NET / NETMax is greater than 0.1, still more advantageously greater than 0.5.
[0047] Advantageously, the pore size of the continuous medium is greater than twice the molecular diameter of the species to be separated. Advantageously, the pore size of the continuous medium is greater than 10 times the molecular diameter of the species to be separated. Advantageously, the pore size of the continuous medium is greater than 2 times and less than 1000 times the molecular diameter of the species to be separated. Advantageously, the pore size of the continuous medium is greater than 2 times and less than 30 times the molecular diameter of the species to be separated. In the condensed phase the steric hindrance to diffusion caused by pores is calculated by the formula (Deen, 1987): With C = Kp * Kr And Kp = (1 - To) 2 Kr = 1 - 2,104 * To + 2,089 * A2 - 0,948 * À3 Rh À = - ro Rh is the molecular radius of the species molecule to be separated as a sphere and the radius of the pores. Kp is a factor that accounts for a difference in the equilibrium concentration between the pores and the infinite medium. Kr takes into account the steric genes of the molecules to be separated in the pore volume.
[0048] This is the diffusivity reduction factor in the free medium to be applied to obtain the diffusivity in the pores. It is thus found that the group C becomes less than 0.1 for a ratio Δ of 0.5, corresponding to a pore size less than 2 times the diameter of the species molecule to be separated. One loses an order of magnitude on the effective diffusivity, which becomes prohibitively low, and the efficiency of the separation becomes bad. The following table calculates the C ratio for different molecules and pore sizes. Molecule rh m ro (n Kp Kr Organic 0.15 0.3 1 0.00 0.04 0.00 Organic 0.15 0.6 0.5 0.25 0.35 0.09 Organic 0.15 1 0 , 3 0.49 0.53 0.26 Organic 0.15 2 0.15 0.72 0.73 0.53 Organic 0.15 4 0.075 0.86 0.85 0.73 Organic 0.15 6 0, 05 0.90 0.90 0.81 Organic 0.15 10 0.03 0.94 0.94 0.88 Protein 1.5 6 0.5 0.25 0.35 0.09 Protein 1.5 10 0 , 3 0.49 0.53 0.26 Protein 1.5 30 0.1 0.81 0.81 0.66 Protein 1.5 100 0.03 0.94 0.94 0.88 macromolecule 5 30 0, 33 0.44 0.50 0.22 macromolecule 5 100 0.10 0.81 0.81 0.66 macromolecule 5 300 0.03 0.93 0.93 0.87 In the gas phase the diffusion becomes impeded when the flow This occurs when the average free path of the molecules becomes of the order of or greater than the pore diameter Advantageously, the packing has a population of related pores whose diameter is greater than the average free path of the molecules to be measured. separating under the conditions of the process.The diffusivity of Knudsen is written : dPore 8 * K * Na, * TK DKA = 3 * * mA When Knudsen's diffusivity and molecular diffusivity are in competition, we write: 1 1 1-a * yA DAe-DKA + DAB With NB = 1 + - NA In general we simplify this formula by: 1 1 1 DAe DKA + DAB The coefficient C deduces CDAe DKA DAB DAB + DKA In these formulas we note: DKA: diffusivity of Knudsen, m2 / s DAB molecular diffusivity m2 / s DAe: medium diffusivity m2 / s TK: absolute temperature, Kelvin MA molar mass of component A, kg / mol K Boltzmann constant, MKSA Nay: number of Avogadro dpore: pore diameter, m In particular we can consider as standard species test, water, hexane or methanol, in the liquid phase or in the gas phase at the saturated vapor pressure at 25 ° C. Effective diffusivity is the experimentally observed diffusivity found in a real material considered as a macroscopic whole. The actual material is composed of the material extending between the conduits with the exception of these. It comprises at least one porous organic gel or an organic liquid (non-porous) and a possible structural material. The diffusivity in the actual material is related: - to a possible pore volume thereof in which the mobile phase can penetrate and in which the species to be separated can diffuse, and - to the diffusivity in the organic gel itself. The porosity of a crosslinked organic gel may result from its swelling in an organic solvent, mineral or aqueous. This is for example the case of styrene copolymers and 2 to 8% of divinylbenzene. It may also come from a porosity of the gel in the non-solvated state. This is for example the case of styrene copolymers and 20 to 80% of divinylbenzene polymerized in the presence of a porogenic solvent such as an aliphatic alcohol having 8 to 12 carbon atoms in its molecule. It may also come from the porosity of a support on which a polymeric gel is deposited in the form of a thin layer.
[0049] The effective diffusion is measured by imposing on both sides of a uniform thickness Eu and a surface S of material representative of the actual material different concentrations Caval and upstream of a molecule dissolved in the stationary phase, and measuring the material flow cl) of this molecule through said resulting thickness.
[0050] The Deff diffusion coefficient is deduced from the formula: Deff = C1). EU / ((Camont-Cavai) .S) The material exchange surface between a duct and an adjacent duct is advantageously greater than 2% of its periphery, more preferably greater than 10% of its periphery, and even more advantageously greater at 20% of its periphery. In practice the ducts are advantageously connected by a continuum or diffusive bridge in the condensed state. In practice, the effective diffusivity of the species to be separated in the walls of the packing is advantageously greater than one thousandth of their diffusion coefficient in the free mobile phase, more advantageously greater than one hundredth of their diffusion coefficient in the free mobile phase, and even more preferably greater than one-tenth of their diffusion coefficient in the free mobile phase. In order to measure the effective diffusivity, the so-called steady-state conductivity cell method is preferably used. An exhaustive description of this method can be found in [3]. In particular, the effective diffusivity will be measured after complete equilibration of the stationary phase contained in the cell and the species on which the measurement is made. In the case of affinity chromatography, this generally amounts to measuring on a stationary phase saturated with adsorbate. In practice it is ensured that the effective diffusivity of the solutes through the walls of the conduits is advantageously maintained above 12 M2 / S in liquid chromatography, above 1 ° -1 m 2 / s in phase chromatography. supercritical, and above 1e-10 m2 / s in gas phase chromatography. The diffusion of the species of the mobile phase into the walls of the packing can be carried out in at least two ways, namely by diffusion into the mass of the gel or organic liquid or by diffusion outside the organic gel, in its porosity. Such diffusivity can be obtained by a combination of porous volume, organic gel crosslinking rate, elution solvent and operating temperature according to the knowledge of those skilled in the art and the available data.
[0051] The molecular diffusion-permeable organic gel used in the present invention can also be defined by the fact that its permittivity with respect to the species to be separated is greater than 5000 Barrer. The Barrer is a unit of permittivity defined in (cm3 of diffusing solute, in the form of perfect gas under the conditions standard.cm) / (cm2 s cmHg): 1 Barrer = 1e-16 (cm3 02 cm) / (cm2 s cm Hg) The table below shows the permittivities of common organic polymers with respect to oxygen and water. Polymer Trade name Permittivity 02, Permittivity H20, Barrer Barrer Poly (isoprene) Natural rubber 23.3 2290 PolyChloroprene Neoprene G 4.0 910 Poly (vinyl chloride) PVC (unplasticized) 0.045 275 Poly (tetrafluoroethylene) Teflon 4.2 4, 8 Poly (ethylene) low LDPE 2.2 68 density Poly (propylene) PP 1,2 35 Poly (methyl methacrylate) Plexiglas 1,2 3200 Poly (carbonate) Lexan 1,4 1400 Unsaturated polyester Polyester 750 Cellulose Cellulose 18900 Indicative , the diffusivity of water is significant only for materials having a permittivity greater than 5000 Barrer (poly isoprene, Plexiglas) and preferably greater than 25000 Barrer (cellulose). On the other hand, a material such as unsaturated polyester has a permittivity comparable to that of polyethylene and polycarbonate, which are materials known to be waterproof and nonporous since they serve to make containers or impermeable walls. Advantageously, the permittivity of the organic gel with respect to the species to be separated is greater than 105 Barrer, and even more preferably greater than 106 Barrer. In order to measure the permittivity of the organic gel, the assembly described in reference [1] will be used. The lining may comprise, as organic gel permeable to molecular diffusion, a gel consisting mainly of organic chemical species and therefore mainly composed of carbon and species usually bound to carbon in the context of organic chemistry. In particular, it will mainly consist of a combination of a carbon atom skeleton and atoms of hydrogen, oxygen, nitrogen, phosphorus, sulfur, chlorine, fluorine, bromine, diode. The hydrocarbon species, halocarbon, hydroxycarbon, oxycarbon, sulfo carbon, phosphorocarbon, nitrocarbon, etc., will be mentioned. of natural or artificial origin. Said gel has, under the conditions of its use, a permittivity sufficient to allow molecules or macromolecules to diffuse into its breast or its pore volume and to interact selectively with it in order to achieve a chromatographic process in less than a few hours, advantageously less than a few tens of minutes and advantageously less than a few minutes. By "conditions of its use" is meant essentially temperature conditions and the use of gaseous mobile phases, liquid or supercritical to take advantage of its permittivity by allowing species (molecules or macromolecules) present in the mobile phase of diffusing into the mass or the pore volume of said gel and selectively interacting with it. In the context of the present invention, an organic gel is an essentially macroscopic notion. Said gel is a material in its own right that can exist and be handled independently of its possible support. This basically distinguishes it from layers of molecular thickness resulting from a grafting of silanes on a surface, for example, which do not exist independently of their support. According to a preferred embodiment of the invention, this organic gel consists of an organic polymer. Advantageously, the molecular weight of this polymer is greater than 1000 g / mol. According to one embodiment, the organic gel is a copolymer of styrene and divinylbenzene. The copolymers of styrene and divinylbenzene exhibit high diffusivity and permeability with respect to molecules dissolved in a solvent. In order to increase this diffusivity, the rate of styrene is decreased. This decrease has the effect of making the polymer mechanically more fragile. However, because of the low pressure drop of a multi-capillary packing, the mechanical forces on the polymer are less important than for a lining without ducts, which makes it possible to reduce the styrene content without degrading the mechanical strength of the packing by compared to known packings without capillary conduits. According to a first embodiment of the invention, the weight content of divinylbenzene in styrene is less than 20%, advantageously less than 8%, and even more advantageously less than 2%. The porosity of such gels develops by swelling with an eluting solvent. They are in particular used in size exclusion chromatography. According to a second embodiment of the invention, the weight content of divinylbenzene in styrene is greater than 20%, advantageously greater than 40%, and the polymerization is carried out in the presence of a pore-forming agent such as a solvent of the monomers in which the polymer is insoluble. The porosity of such gels is inherent to their structure in the dry state. The gels of polyvinyl alcohol, polymethyl methacrylate, polyhydroxymethyl methacrylate, polyacrylamide, hydroxyethyl methacrylate copolymerized with dimethacrylate may also be mentioned in a nonlimiting manner as being used for carrying out the invention and as materials constituting the lining. glycidyl (GMA-EDMA), etc. The polyacrylamides advantageously consist of copolymers of acrylamide and N N 'methylenebisacrylamide.
[0052] Mention may also be made of cellulose (a polyholoside) and its derivatives, especially carboxymethyl celluloses and diethylaminoethyl celluloses for ion exchange chromatography. It is also possible to use organic polyloside gels known in the state of the art using organic macromolecules such as crosslinked dextrans, for example by N, N methylened diacrylamides or epichlorohydrins. These gels are in particular known under the trade names Sephacryl TM and SephadexTM, products of GE Healthcare. It is also possible to use other organic polysaccharide gels using organic macromolecules such as crosslinked agaroses, for example by epichlorohydrin. These gels are in particular known under the trade names Sepharose TM and SuperdexTM, Superose TM products from GE Healthcare. Other organic gels can also be used using artificial macromolecules such as vinyl polymers containing many hydroxyl groups. These gels are in particular known under the trade name ToyoPearin®, products of Tosoh group, in particular those organic gels can be made from mixtures of monofunctional monomers and multifunctional monomers The multifunctional monomers crosslink the polymer obtained. Organic compounds may be prepared from mono, di and multifunctional monomers known in the art, which may be monomers containing epoxy, vinyl or hydroxyl siloxane radicals, and may be styrene and its derivatives containing hydroxyl, halogen, amino, sulphonic, carboxylic, NO2 group, C4, C8, C12, C18 alkyl chains, etc. These monomers can be acrylates, methacrylates, acrylamides, methacrylamides, vinylpyrrolidones, vinylacetates, acrylic acid, and the like. methacrylic acid, vinyl sulfonic acid The siloxanes may include a hydroxyl group, vinyl, alkyl groups, etc. ...
[0053] In particular these monomers may be chloromethyl styrene, 4-acetoxystyrenea, methacryl methyl, ethyl, propyl, butyl, hexyl, lauryl, triphenylmethyl, pyridyl-2 diphenylmethyl, acrylate methyl, ethyl, propyl, butyl, hexyl, lauryl, glycidyl methacrylate, AMPS, 2-vinyl-4,4-dimethylazlactone, epoxypropyl 2-3 methacrylate etc. .
[0054] These functional groups can be provided before or after the polymerization. The monofunctional monomer level may vary between 2% and 98% by weight of the total monomers. Advantageously it is between 2% and 40% by weight of the total monomers. The bi or multifunctional monomers may be monomers based on benzene, naphthalene, pyridine, alkyl ethylene, glycol, etc. having two or more vinyl or epoxy functional groups. Examples of these components are divinyl benzenes, divinyl naphthalene, alkyl diacrylates and dimethacrylates, diacrylamides, and dimethylacrylamides, divinyl pyridines, dimethacrylates or diacrylates of ethylene glycol, polyethylene glycol, di, or trimethylolpropane trimethacrylate, 1,3 butanedioldiacrylate, di, tri or tetra pentaerythritol methacrylates or acrylates, or mixtures thereof. Hydroxyl, tri or tetra siloxanes often generated from alkoxysilanes can be used. The level of bi or multifunctional monomer may vary between 100% and 2% by weight of the total monomers.
[0055] Advantageously, said content is between 98% and 60% by weight of the total monomers. The initiators used for the polymerization include all those included in the state of the art, such as azo compounds and peroxides. Typical examples are azobisisobutyronitrile, benzoyl peroxide. The typical amount of initiator generally ranges from 0.4 to 2% by weight based on the weight of the monomers. In the case of siloxanes, acid hydrolysis is preferred. Advantageously, these mixtures are polymerized in the presence of a porogenic agent subsequently removed as an organic solvent or a non-reactive polymer. These porogens have been mentioned previously. In particular, dodecanol-1 and cyclohexanol-1 are mentioned. The amount of pore-forming agent may vary between 10 and 90% and preferably between 20 and 60% by volume of the final mixture comprising the monomers.
[0056] X-ray or gamma-ray polymerization can be used for the homogeneous production of large parts. Advantageously, these organic gels can be surface-treated in order to functionalize them by grafting, for example by means of silanes. They may be provided with sulphonated groups (sulphonation), quaternary ammoniums, octadecyl, octyl, butyl, phenyl, amine, diethylamino, ethyl, sulphopropyl or carboxymethyl radicals, hydrophilic polymers, alpha, beta, gamma or methylated cyclodextrins. no, L-amino acids or D-amino acids, proteins, etc. On gels containing aromatic rings functionalization reactions such as chloromethylation, amination, nitration, sulfonation, alkylation may be used. and Friedel Crafts acylation, etc. Advantageously, the organic gels that can be used for ion exchange or complexation chromatography comprise aminodiacetate, phosphonate, amidoxime, amidophosphonate, thiol, sulfonate, primary amine, secondary or tertiary amine or quaternary amine radicals. Advantageously, the organic gels that can be used for the chromatography of complex donor acceptors of elec tron include nitro or chloroaromatic radicals, or phenoxy, pyrene, quinazoline-2 radicals, such as dinitro-2-aminopropyl, trinitro 2,4,6-anilinopropyl, tetranitro-2,4,7-fluorenoiminopropyl, tetrachloro phthalimidopropyl, nitro- 3-naphthalimido-propyl, naphthalene tetracarboxy-1,4,5,8-diimidopropyl, pyromellidiimidopropyl, pentafluorobenzamidopropyl, caffeine, phenoxy, quinazolin-2, quinolinol, -8,2,2,2-trifluoro-1 (10-methyl) -9anthryl) ethanol, pyrenepropyl, etc. Advantageously, the organic gels that can be used for ligand exchange chromatography include the bis dithiocarbamate, cyclam, oxine, dialkyldithiocarbamate, and the like radicals. Advantageously, the organic gels that can be used for the separation of chiral molecules comprise chiral selectors generally chosen from Pirkle type groups, cyclodextrins and crown ethers, natural and synthetic polymers, and proteins. Among the Pirkle-type groups, mention will be made, inter alia, of: R or S (dinitro-3,5) benzoyl) phenylglycine R or S N- (dinitro-3,5-benzoyl) tyrosine n-butylamide S-N- (dinitro- 3,5 benzoyl) tyrosine (naphthyl) -1 ethylamide R or S - (3,5-dinitro-benzoyl) phenylglycine S - (dinitro-3,5-benzoyl) leucine S- (dinitro-3,5-benzoyl) phenylalanine R or S-Naphthylamine R α-methylbenzylurea S α- (naphthyl) -1-ethylamine The following derivatives are also mentioned: R-phenylglycine and S- (4-chloro-phenyl) -isovaleral acid R-phenylglycine and 1-R-acid, 3-R, chrysanthemic Dinitro-3,5-benzoyl and R or S 1-naphthyl glycine tert-butylamine and S valine Dinitro-3,5-aniline and S valine Dinitro-3,5-aniline and S-tert-leucine S - (α-naphthyl) -1 ethylamine and S-valine R- (α-naphthyl) -1 ethylamine and S-valine R-phenylglycine and R- (α-naphthyl) -1 ethylamine R-phenylglycine and S- (a- naphthyl) -1 ethylamine S-proline and R- (α-naphthyl) -1 ethylamine S-proline and S- (α-naphthyl) -1 ethylamine S-tert-leucine and R- (a-naphthyl) -1 ethylamine S-tert-leucine and S- (a-naphthyl) -1 ethylamine tartaric acid and dinitrobenzylphenyl-ethylamine Among the ligand exchangers include among others: Proline, hydroxyproline, valine, etc ... Carboxymethylamino-2 -diphenyl-1,2-ethanol Among the cyclodextrins and crown ethers, mention may be made, inter alia, of the following: α, β, γ-cyclodextrin α, β, γ-substituted cyclodextrin cyclodextrins (β-hydroxypropyl) β-cyclodextrins derived (hydroxypropyl) racemic) f3-cyclodextrin derivatives (S or R (α-naphthyl) -1 ethylcarbamate) f3-cyclodextrin derivatives (racemic ((a-naphthyl) -1 ethylcarbamate) f3-cyclodextrin derivatives (dimethyl-3,5-phenylcarbamate) f3-cyclodextrin derivatives (para toluyl) Among the natural polymers, mention may be made of, inter alia: Triacetylated microcrystalline cellulose Triacetate of cellulose Tribenzoate of cellulose Tri cellulose phenylcarbamate Cell tri- (dimethy1-3,5-phenyl) carbamate dyes cellulose tri-4-chlorophenylcarbamate cellulose tri-4-methylphenylcarbamate cellulose tri-4-methylbenzoate cellulose tricinnamate Tri- (phenylethylamine) amylose carbamate Tri- (dimethy1-3,5-phenyl) amylose carbamate the synthetic polymers include, inter alia: Poly (N-acryloyl-1-phenylalanine ethylester) Poly (triphenyl methylmethacrylate) Poly (pyridyl-2-diphenyl-methylmethacrylate) Among the proteins, mention may be made, inter alia, of: Serum albumin (BSA) Α-glycoproteic acid Human serum albumin Ovomucoid.
[0057] A list of these groups can be found in reference [2] p.574 and 575. These organic gels can be used to perform affinity chromatography. Affinity chromatography separates biochemical molecules based on highly specific interactions such as antigen-antibody, enzyme-substrate, or receptor-ligand.
[0058] The contacting is done by circulation of the mobile phase through the conduits of the packing, allowing the attachment of the target molecules on the organic gel. The bound compounds are eluted by a change in pH, pH, salt concentration, charge, or ionic strength in general. This proceeds in one step (slot) or by an elution gradient to recover the molecules of interest.
[0059] Various forms of affinity chromatography are usually considered: by immunoaffinity, by immobilized metal ions, by recombinant proteins, by lectins. In particular, by immunoaffinity, proteins are coupled to a substrate such as agarose covalently, and they are used for the purification of their antibodies.
[0060] In particular, by immobilized metal ions, metal ions such as Cu, Ni, Co are coupled to a substrate such as agarose for the purification of proteins or peptides containing histidine. Metallic ions such as Fe, Zn, Ga are coupled to a substrate such as agarose for the purification of phosphorylated proteins or peptides. Elution is by pH change or by solutions of competitors such as imidazole.
[0061] In particular, by means of recombinant proteins, the proteins are labeled, so as to be able to select them, by markers such as glutathione S transferase (GST, hexahistidine (Hs), Maltose (MSP).) Histidine has a high affinity for Ni or Co by coordination or covalence Elution is by solutions containing an excess of solute capable of binding to the immobilizing metal, such as imidazole, or as for example an excess of glutathione. for lectins, the labeling of the molecules allows them to bind selectively to carbohydrates.Ligands such as conconovalin A bind to glucose chains glycoproteins and can isolate them.The packing containing organic gels according to the invention are suitable These gels are characterized by a high unit volume cost, it is advantageous to reduce the immobilized volume for a given production.
[0062] Advantageously, the packings for affinity chromatography have a small volume of ducts in front of the total volume of the packing, preferably less than 40%, more preferably less than 20% and even more preferentially less than 10% of the total packing volume. . Advantageously, the distance between adjacent ducts is less than 0.5 mm, preferably less than 0.25 mm, and still more preferably less than 0.1 mm, in order to ensure sufficient rapidity of the diffusional processes. The propagation front of the retained molecule is thus steeper and short column lengths can be used, further reducing the volume of the packing. Advantageously, the organic gel represents a high fraction of the volume of the packing considered, excluding the volume of the ducts, advantageously greater than 40%, more preferably greater than 60% thereof. Advantageously, these packings are used for affinity chromatography in radial or axial continuous annular chromatography devices. This minimizes the amount of stationary phase required for a given production by reducing the amount of stationary phase immobilized by means of short cycle times without expensive devices using instruments operating under low pressure. According to one embodiment of the invention, a porous polymeric organic gel may comprise in its porosity a third solid body. Advantageously, in this case, the lining comprising the polymeric organic gel and intended to serve as a support for a solid third body will have related pores of high size, greater than 50 nm, preferably greater than 200 nm, and even more preferably greater than 500 nm. . Advantageously, the lining comprising the polymeric organic gel and intended to serve as a support for a solid third body has a specific surface area of less than 20 m 2 / g. Advantageously this solid third body is a stationary phase for chromatography. Advantageously this third solid body may be any of the previously listed stationary phases.
[0063] Advantageously this third solid body can be a stationary phase mineral or organo metal. Among these will be named in a nonlimiting manner the oxides of silicon, alumina, titanium, zirconium. Advantageously, this solid third body may be a high surface area silica for chromatography. More generally, the invention relates, in the field of liquid chromatography: liquid-liquid partition chromatography, partition chromatography on grafted stationary phases and nonionic polymers, exchange chromatography, ion chromatography, ion pair chromatography, ligand exchange chromatography, electron donor-acceptor chromatography, steric exclusion chromatography, all variants affinity chromatography. Advantageously, the injection of the mobile phase is through a filter whose cutoff diameter is smaller than the diameter of the ducts, and preferably ten times smaller than the diameter of the ducts. Figure 1 is a sectional view of a multicapillary packing 3 according to one embodiment of the invention in a plane parallel to the longitudinal axis of said lining. In this embodiment, the lining is a porous monolith formed of an organic gel 2 traversed by capillary ducts 1 where a fluid or supercritical mobile phase passing through the lining 3 can flow freely. By "monolith" is meant a porous material in one piece, comprising a continuous skeleton, which can be made of one or more materials. In the example illustrated in FIG. 1, the capillary ducts are straight, parallel, and evenly spaced. The different ducts have morphologies and diameters as identical as possible. Each duct passes through the lining from one side to the other, that is to say it advantageously has its ends open on each side 4 and 5 of the lining, allowing the flow of the fluid from the inlet side 4 to the outlet side 5. Such packing can therefore be used in a chromatographic column. Figure 2 is a top view of the face 5 of the lining of Figure 1 seen in the direction 6. There are the openings of the individual capillary ducts 1 in the mass 2 of organic gel. According to an alternative embodiment, insofar as the organic gel generally has a limited mechanical strength, the monolith comprises a porous skeleton made of a dimensionally stable and more resistant material than the organic gel, and the pores of said skeleton are covered at least partly organic gel. FIG. 3 thus represents a skeleton 7 made of a dimensionally stable material, obtained for example by sintering a molded or extruded powder according to the form of the packing, or obtained by a sol gel or filler-binder process. The organic gel 8 is deposited in the form of a thin layer on the surface of the skeleton 7, and in particular on the surface of the pores. The lining has a continuous network of macropores 9 allowing molecules to diffuse rapidly throughout the thickness of the lining and between adjacent ducts. Figure 4 illustrates another embodiment of the lining. According to this embodiment, hollow fibers are juxtaposed according to an optimal stack that is as compact as possible. These hollow fibers consist of an empty core 101 in which the mobile phase flows and a porous envelope 100 retaining the organic gel or consisting of the organic gel. The space between the fibers 102 is filled with either the mobile phase or the gel or organic liquid. Various methods of manufacturing a packing as described above will now be described. According to a first embodiment of the invention, it is possible to coat an organic gel with a multicapillary monolith obtained by extrusion and sintering. This monolith must be porous. It must therefore be prepared and sintered under conditions allowing to maintain a significant porosity. However, the diameter of the ducts obtained by such an extrusion process, which is of the order of one millimeter, is generally too high with regard to a chromatographic application.
[0064] According to one embodiment, to form a monolithic packing, a method is implemented comprising the steps of: - providing a bundle of son called "precursors conduits", that is to say, son that are intended to be eliminated later to leave their footprint in the form of ducts. Advantageously, the outer diameter of the wires is equal to the inside diameter of the ducts - formation of a porous matrix around the wires or ducts, - removal of the wires so as to form said capillary ducts. According to one embodiment, the matrix formed around the son or conduits consists of a polymeric organic gel.
[0065] Alternatively, the matrix formed around the son or conduits is loaded with an organic gel.
[0066] Optionally, the precursor leads of the conduits comprise an ablative layer of a coating material removed during a first step of the son removal treatment. This ablative material may in particular be a wax or a paraffin. Commercial fibers of nylon, polypropylene, cellulose acetate, polyester, aramid, carbon fiber, metal, etc. can be used as precursors son of these conduits with such constraints. Fibers containing no toxic metal salts are preferably used in order not to pollute the final structure of the material. According to a particular embodiment of the invention, the precursor son of the ducts, possibly coated with the ablative layer, are coated with a spacer prior to the formation of the bundle so as to ensure a minimum thickness of material between two adjacent ducts. . According to one embodiment, son consisting of a hydrolyzable polymer are assembled in a bundle, the bundle is immersed in a precursor solution of a porous organic gel, solution which is caused to gel around the son, and the son are removed by hydrolysis to low molecular weight soluble species by dipping the packing in an acidic or basic solution. Indeed, the very inert nature of an organic gel makes it possible to put it in contact with strongly acidic and strongly basic aqueous solutions. By "precursor solution of an organic gel" is meant a liquid of composition such that, by its evolution under the conditions of the manufacturing process, it leads to an organic gel. Among the hydrolysable polymers, mention may be made, inter alia, of polyesters derived from glycolic acid, lactic acid, cellulose, and in particular cellulose acetate, polyglycolic acid or its copolymers with lactic acid, with ε-caprolactone or with trimethylene carbonate, as well as polydioxanone. A polymer is preferably chosen, the hydrolysis of which is rapid at a temperature of 80 to 100 ° C. in an acid medium or, preferably, a basic medium. According to another embodiment, a multicapillary monolith is produced in silica gel. The porous fraction external to the ducts of the monolith receives the organic gel. The silica backbone is removed by dissolution in an aqueous solution at pH greater than 11.0. In this case, the silica gel acts as a blowing agent. According to one embodiment of the invention the son consist of a metal or alloy of low melting point metals and removed by melting and flow out of the material.
[0067] Advantageously, a metal alloy whose melting temperature is lower than the degradation temperature of the material constituting the organic gel is chosen. Preferably, metals having a melting point of less than 220 ° C., preferably less than 150 ° C., and even more preferentially less than 100 ° C., are chosen. A major use of these organic packings is to make separations on molecules and liquids intended for human or animal consumption. In particular, it concerns drinking water, medicines, food additives, etc. In such a case, all the elements and components of the packing must be compatible with strict health constraints. In particular it is important to avoid any pollution of the packing by toxic residues of the manufacturing process, and to avoid as much as possible the use of toxic manufacturing intermediates. The materials or constituents of the precursor metal son of the conduits are part of these intermediates. In fact, residues of these metals can remain in the packing and pollute the species which will be treated there, or to spread in the nature after its destruction. Among the low melting point metals are mainly lead, tin, bismuth, gallium, mercury, silver, cadmium and indium. Since lead, cadmium and mercury are harmful heavy metals that are harmful to human and animal health and the environment, alloys that do not contain these elements and based on tin, bismuth, indium, gallium, silver or any combination thereof with each other or with other less fusible metals. In particular it may be a mixture of bismuth and tin. There is in particular a eutectic mixture of these metals comprising 58% by weight of bismuth, 42% by weight of tin, and melting at 138 ° C.
[0068] Alternatively, it may be metal alloys based on indium. Of these indium alloys, a 52% indium alloy and 48 wt% tin are preferably used, melting at 118 ° C. It is also possible to use a 32.5% Bismuth, 51% indium and 16.5% by weight tin alloy, melting at 60 ° C. The matrix can be created by a charge-binding process. Advantageously in this case it can be created from a mixture containing a solid third body as a small particle size charge and a binder such as a soil, suspended in a liquid phase. This soil can be any organic or mineral soil, silica sol, alumina, titanium, zirconium, natural or artificial latex, soil of various polymers, colloids. It will be noted for example - natural latex floors, - polystyrene latices, and their functionalized derivatives (amino, carboxy, etc.) - latex copolymers of styrene with butadiene, acrylic acid, and their derivatives functionalized, - soils of nitrile rubbers, etc ... - Soils of polymethylmethacrylate It can also be a soil created in situ by a sol-gel process. The binder may also be a colloid such as albumin, dextran, gelatin, hydroxyethylated starches, etc. Advantageously, the binder will not penetrate the pores of the filler. This can be obtained for example by choosing a binder divided into particles of larger size than those of the pores. In particular, the multicapillary packing supporting the organic gel may be created by a sol gel method. Without departing from the scope of the invention this sol gel method may also be based on an aluminosilicate such as a clay for example.
[0069] Without departing from the scope of the invention, this sol-gel process may also be based on a zirconium oxide, a titanium oxide, a rare earth oxide such as yttrium, cerium or lanthanum, a boron oxide, an oxide of iron, calcium magnesium oxide of strontium or barium, germanium oxide, phosphorus oxide, lithium, potassium or sodium oxide, Niobium oxide or copper oxide. These compounds may be the base of the gel or be combined with one another so as to create a multicomponent gel. Advantageously, as the base of the gel, gels of zirconium oxide or of titanium oxide may be used. Advantageously, the sol-gel process leading to these mono or multicomponent gels will be based on the hydrolysis of organometallic compounds such as alkoxides of the metals in question, alone or mixed with other organometallic compounds and optionally with metal salts such as nitrates or chlorides. . Another method for producing the lining according to the invention comprises the use of hollow fiber bundles with porous walls impregnated with organic gel by dipping and then drained so as to release the lumen of the fibers. It is thus possible to manufacture large packings. Such a manufacturing method comprises the steps of: - providing a compact bundle of hollow fibers, - inclusion in the porous wall of the hollow fibers of an organic gel or its polymerized precursor in situ, so as to leave free and open the lumen of the hollow fibers, - creation of porous or liquid material diffusing bridges between the hollow fibers.
[0070] Diffusion bridges are understood to mean a continuum of molecular diffusion permeable material extending between the fibers. The compactness of the stack of hollow fibers makes it possible to reduce the molecular diffusion distances between them.
[0071] Material bonds between the hollow fibers favorably provide effective diffusive bridges between the conduits. The hollow fibers are advantageously made integral with each other so as to reinforce their mechanical cohesion and to handle them collectively. In particular they can be woven into a composite fabric or stick together together in sheet form. In particular, they can be glued together together to form a monolith. The binder may be the organic gel itself. According to one embodiment, it is possible to immerse the external volume at the periphery of the fibers in an organic liquid.
[0072] According to one embodiment, it is possible to use as binder fibers between them a resin other than the organic gel, porous binder resin. This latter method can be combined with a reinforcement by a fabric of structural fibers. According to another method for producing the packing according to the invention, the organic gel is molded into a structure defining conduits. Advantageously, this molding is performed so as to obtain an organic gel film having a spacer such as reliefs. The walls of the ducts may constitute this spacer. The molding can be performed by stamping, extrusion, calendering of a pre-existing film. This process can be carried out alone or on a film or reinforcing fabric.
[0073] The method may comprise polymerizing the organic gel in situ in the mold having the preform of the spacers and then demolding. Alternatively, the process comprises molding the molten organic gel or its molten precursor in the mold and solidifying it, and then demolding it. The film is then stacked or rolled to obtain the final packing.
[0074] The spacers provide free passage of fluid through the structure. When a porous polymeric organic gel contains a solid third body, said third body may be created by dipping the multicapillary lining into a formulation containing the third body in suspension or a precursor of the third body, or both simultaneously followed by drainage and drying of the gel.
[0075] In particular the third solid body may be created by a charge-binder process. Advantageously in this case it may be created from a mixture containing a solid third body as a small particle size charge consisting of a stationary phase for the particle size chromatography less than the pore size of the polymeric organic gel and a binding as a sol, suspended in a liquid phase impregnating the porosity. This soil may be an organic or inorganic soil, silica sol, natural or artificial latex, soil of various polymers, etc. It may also be a soil or a gel created in situ by a sol-gel process.
[0076] In particular the solid third body may be created on the surface of the polymeric gel by impregnating it with a colloidal solution optionally followed by its gel, and its drying. Silica sols with a high specific surface area are particularly suitable. In particular the third solid body may be created on the surface of the polymeric gel by a sol gel process. Any sol gel method as described previously in this specification may be used. In particular it may be a bimodal silica obtained by a sol gel process. Advantageously this third solid body is silica for high surface area chromatography bound by a silica sol.
[0077] Figures 5 and 6 are seen from above and seen in section respectively an organic gel 10 traversed by conduits 11. This organic gel is in the form of sheet obtained by molding on a preform. It can be attached to an underlying fabric 12 providing better strength. The organic gel may contain a reinforcing filler. The leaves can be rolled up or stacked to achieve any desired shape. Advantageously, the organic gel or the third body that it supports is functionalized after the creation of the conduits. By functionalization is meant the addition of particular chemical groups conferring reactivity or selectivity to the raw packing. It is in particular the contribution of reactive groups ion exchangers, treatment with silanes, grafting of polysaccharides, proteins and chiral molecules, etc. The packing typically has a section greater than 20 cm 2, and preferably greater than 100 cm 2. The lining may be in the form of a disk or a column having two flat straight sections, encapsulated in a contiguous envelope serving as mechanical protection. The tightness of the junction between the lining and the casing can be ensured by an adhesive, a polymer or a plastic film, in particular a heat-shrinkable film. Advantageously, the lining can be created directly in said envelope.
[0078] A chromatographic process is generally characterized by the effectiveness of the packing expressed in number of theoretical plates. Advantageously, the packing has an efficiency greater than 1000 theoretical trays per meter of packing, preferably more than 10,000 theoretical trays per meter of packing and more preferably more than 100,000 theoretical trays per meter of packing. FIG. 7 compares the efficiencies of a multicapillary packing with respect to a chromatographic separation when the walls separating the capillary ducts are porous or non-porous, obtained by computer simulation. The abscissa axis represents the length of the packing expressed in micrometers. The axis of ordinates represents the effectiveness of the packing expressed in number of theoretical plates (NPT).
[0079] The diameters of the ducts are distributed according to a Gaussian law. Curve (a) represents a packing consisting of ducts of randomly varying diameter according to a Gaussian statistical law around an average of 10 μm with a standard deviation of 0.5 μm, for non-porous walls. Curve (a) shows the effectiveness of such packing whose walls are non-porous and whose capillaries behave therefore independently of each other. This efficiency begins with increasing then capping to tend towards a limit independent of the length of the lining. This phenomenon is due to the fact that the diameters of the capillaries are not uniform but distributed according to a random Gaussian law.
[0080] Curve (b) represents the same beam with porous walls having 55% porous volume, a wall thickness of 2 μm and a pore size ten times greater than the molecular diameter of the species to be separated. Curve (b) thus shows the effectiveness of a packing of the same dimensions as the previous one but whose walls of the ducts are porous and in which the adjacent capillary ducts communicate by molecular diffusion. In this case, the efficiency no longer caps but increases proportionally to the length of the lining despite the same Gaussian distribution of the diameters. The non-uniformity of behavior of these conduits is leveled by molecular diffusion between them.
[0081] This phenomenon is specifically relevant for a chromatographic process, where high efficiencies are required. It should be noted that such properties are of secondary importance for an adsorption or catalysis process, and of no interest for a filtration process. Figures 8 and 9 describe the diffusive flux considered to define the concept of molecular permeability according to the invention. FIG. 8 describes the molar flow rate of diffusion between the adjacent ducts under the effect of a given concentration difference of the species to be separated between the walls of said ducts. The duct 13 is assumed to have a concentration profile dictated by the hydrodynamic conditions of the flow resulting in an average Ce concentration. Adjacent ducts are assumed to have a Cs concentration lower here than Ce. The average diffusion molar flow rate Phip is constituted by the sum of the flows (15, 15 ', 15 ", 15") leaving the periphery of the duct 13 and passing through the lining 14. The medium 14 consists of porous organic gel, liquid organic and mobile phase, and a possible porous support thereof. FIG. 9 describes the molar flow rate Phic average diffusion between a pipe and the stationary phase that constitutes the lining under the effect of the same difference in concentration of species to separate between the ducts and the wall of said ducts. The molar diffusion rate between a conduit 13 and the stationary phase constituted by the packing is measured through a peripheral zone of the duct delimiting the empty capillary in which the fluid flows, under the effect of the same difference in concentration. species to separate. The exchange is calculated on the basis of a mean concentration Ce of the flow of eluent in the conduit. The periphery of the duct is assumed at a concentration Cs. The molar diffusion rate is constituted by the sum of the flows (16, 16 ', 16 ", 16") passing from the central zone of the duct 13 to the packing 14. The concentrations are expressed in moles / m 3. The examples developed below describe various processes for manufacturing a packing for carrying out a chromatography process according to the invention. Example 1: Manufacture of a multicapillary packing in an anion exchanger The starting material is a wire made of a tin and bismuth alloy in proportions 58%, 42% by weight respectively. It has 15/100 mm of diameter. The wire is cut into rectilinear sections 200 mm long, covered by dipping with a thin layer of a mixture of styrene, 8% by weight of divinylbenzene by weight, 0.4% by weight of an activator of polymerization (azobis isobutyronitrile) and crushed glass powder at 10 μm (one volume of glass powder for one volume of the solution). The sections of wire are arranged for 24 hours at 70 ° C. under nitrogen. They are then introduced into a bundle about 4 mm in diameter in a glass tube 160 mm long and 4 mm inside diameter previously prepared. A mixture of styrene, 8% by weight of divinylbenzene and 0.4% by weight of a polymerization activator (t-butyl hydroperoxide) is then poured into the tube through the interstices between the sections of yarn so as to completely fill this empty space. The mixture is polymerized for 24 hours at 120 ° C. The composite thus produced is released by cutting the sections of wire on either side of the glass tube flush with its ends, perpendicular to said sections.
[0082] The composite is carried in an oven at 145 ° C, until the sections of wire are melted, and the molten metal is removed by ultrasonic vibrations and under the action of gravity and a slight air flow. The monolith thus produced is converted into a basic ion exchanger by subjecting it to a 15% by weight solution of tin tetrachloride in chlorodimethyl ether at 0 ° C. for 6 hours. The lining is then washed with methanol and then with water and quaternized by the action of a 40% aqueous solution of trimethylamine for a period of ten hours. The packing is then washed to neutrality and the quaternary ammonium is converted to its hydroxyl form.
[0083] Example 2 Manufacture of a multicapillary packing in a cation exchanger A monolith with porous walls of alpha alumina 160 mm long having 100 conduits of 0.45 mm in diameter distributed in a square mesh of 1.2 mm side is produced by extrusion and sintering of a 20 μm elemental diameter alpha alumina powder.
[0084] A mixture of 49 g of styrene, 1 g of divinylbenzene, 200 mg of a polymerization activator (t-butyl hydroperoxide) and 150 ml of pentane is then poured into the monolith so as to completely fill the porosity of the wall. The core of the ducts is drained from the liquid phase that it contains. The monolith heated to 50 ° C so as to evaporate the majority of the pentane, then briefly pulled under vacuum so as to complete this evaporation.
[0085] The deposition of monomers and activator thus deposited in the form of a liquid film covering the wall of the grains of alumina is polymerized for 24 hours at 120 ° C. under a nitrogen atmosphere. The monolith thus produced is converted into an acidic ion exchanger by sulfonation. The monolith is treated with a stream of concentrated sulfuric acid containing 0.1% by weight of silver carbonate at 100 ° C for 3 hours. The sulphonated packing is then treated with progressively less concentrated sulfuric acid and finally with distilled water. Example 3: Manufacture of multicapillary packing having porosity A mixture is prepared containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and degassed under nitrogen for 20 minutes. This mixture is heated at 70 ° C. for 24 hours. The mixture polymerizes. The monolith A thus produced is washed and percolated with THF for 30 minutes and dried in an oven at 90 ° C.
[0086] Monolith A is milled under liquid nitrogen until a grain diameter of 10 μm is obtained. Example 3 A yarn of a mixture of indium and tin in weight proportions 48, 52 melting at 118 ° C, is produced with a diameter of 0.25 mm. The wire is cut into rectilinear sections 15 cm long, covered by dipping with a thin layer of a mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and a volume of the monolith A milled at 10 pm equivalent to 9/10 of the volume of the solution. The sections are polymerized for 24 hours under nitrogen at 70 ° C. They are then cut into lengths of 120 mm and assembled into a bundle of about 4 mm diameter in a glass tube 100 mm long and 4 mm in diameter previously prepared.
[0087] A mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile and 120 g of dodecanol is prepared in parallel and degassed under nitrogen for 20 minutes. The previously prepared bundle is filled with this mixture and heated to 70 ° C for 24 hours. The mixture polymerizes.
[0088] The composite thus produced is released by cutting the sections of wire on either side of the glass tube flush with its ends, perpendicular to said sections. The composite is carried in an oven at 125 ° C, until the sections of wire are melted, and the molten metal is easily removed by applying ultrasonic vibrations by gravity and under a slight flow of air under pressure.
[0089] The monolith thus produced is washed under THF for 30 minutes. Example 4: Manufacture of multicapillary packing with porosity A mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and degassing under nitrogen for 20 minutes is prepared.
[0090] This mixture is brought to 70 ° C. for 24 hours. The mixture polymerizes. The monolith A thus produced is washed and percolated with THF for 30 minutes and dried in an oven at 90 ° C. Monolith A is milled under liquid nitrogen until a grain diameter of 10 μm is obtained.
[0091] A polydioxanone wire is produced with a diameter of 0.25 mm. The wire is cut into rectilinear sections 15 cm long, covered by dipping with a thin layer of a mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and a volume of the monolith A milled at 10 pm equivalent to 9/10 of the volume of the solution. The sections are polymerized for 24 hours under nitrogen at 70 ° C. They are then cut into lengths of 120 mm and assembled into a bundle of about 4 mm diameter in a glass tube 100 mm long and 4 mm in diameter previously prepared.
[0092] A mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 890 mg of azobisisobutyronitrile and 120 g of dodecanol is prepared in parallel and degassed under nitrogen for 20 minutes. The previously prepared bundle is filled with this mixture and heated to 70 ° C for 24 hours. The mixture polymerizes. The composite thus produced is released by cutting the sections of wire on either side of the glass tube flush with its ends, perpendicular to said sections. The monolith thus produced is washed by percolating THF for 30 minutes and dried in an oven at 90 ° C.
[0093] The polydioxanone son are dissolved in 90 ° C sodium hydroxide percolated through the packing for 1 hour, then the packing is washed with distilled water until neutral. EXAMPLE 5 Manufacture of Multicapillary Lining in Crosslinked Agarose A mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile and 120 g of dodecanol was prepared and degassed under nitrogen for 20 minutes. This mixture is brought to 70 ° C. for 24 hours. The mixture polymerizes. The monolith A thus produced is washed and percolated with THF for 30 minutes and dried in an oven at 90 ° C.
[0094] Monolith A is milled under liquid nitrogen until a grain diameter of 10 μm is obtained. A polydioxanone wire is produced with a diameter of 0.25 mm. The wire is cut into rectilinear sections 15 cm long, covered by dipping with a thin layer of a mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and a volume of the monolith A milled at 10 pm equivalent to 9/10 of the volume of the solution. The sections are polymerized for 24 hours under nitrogen at 70 ° C. They are then cut into lengths of 120 mm and assembled into a bundle of about 4 mm diameter in a glass tube 100 mm long and 4 mm in diameter previously prepared.
[0095] A mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile and 120 g of dodecanol is prepared in parallel and degassed under nitrogen for 20 minutes. The previously prepared bundle is filled with this mixture and heated to 70 ° C for 24 hours. The mixture polymerizes.
[0096] The composite thus produced is released by cutting the sections of wire on either side of the glass tube flush with its ends, perpendicular to said sections. The monolith thus produced is washed by percolating THF for 30 minutes and dried in an oven at 90 ° C.
[0097] The polydioxanone son are dissolved in 90 ° C sodium hydroxide percolated through the packing for 1 hour, then the packing is washed with distilled water until neutral. One deciliter of agarose beads are dissolved in one deciliter of deionized water at 95 ° C. The porosity of the monolith is impregnated with this solution by soaking and drainage at 95 ° C of the core of the pipes and cooling. The occluded and frozen agarose is washed with distilled water A decilitre of 1N NaOH solution containing 2 ml of epichlorohydrin and 0.5 g of NaBH4 is prepared. We fill the ducts of the monolith. The whole is heated to 60 ° C for one hour. The crosslinked gel obtained is washed with hot water until neutral. EXAMPLE 6 Manufacture of Multicapillary Lining in Crosslinked Agarose A monolith with porous walls of alpha alumina of 300 mm in length and having 100 conduits of 0.45 mm in diameter distributed in a square mesh of 1.2 mm on one side is produced by extrusion and sintering of a 20 μm diameter alpha alumina powder A deciliter of agarose beads are dissolved in a decilitre of deionized water at 95 ° C.
[0098] The porosity of the monolith is impregnated with this solution by soaking and drainage at 95 ° C of the core of the pipes and cooling. The occluded and frozen agarose is washed with distilled water A decilitre of 1N NaOH solution containing 1 ml of epichlorohydrin and 0.25 g of NaBH4 is prepared. We fill the ducts of the monolith.
[0099] The whole is heated to 60 ° C for one hour. The crosslinked gel obtained is washed with hot water until neutral. EXAMPLE 7 Manufacture of Multicapillary Lining in Crosslinked Agarose A monolith with porous walls of 150 mm long pyrex glass having 100 conduits of 0.45 mm in diameter distributed in a square mesh of 1.2 mm on one side is produced by extrusion and sintering of a pyrex glass powder of 20 μm in elemental diameter A deciliter of agarose beads are dissolved in a deciliter of deionized water at 95 ° C. The porosity of the monolith is impregnated with this solution by soaking and draining the pipes at 95 ° C. and then cooling. The occluded and frozen agarose is washed with distilled water A decilitre of 1N NaOH solution containing 20 ml of epichlorohydrin and 5 g of NaBH4 is prepared. We fill the ducts of the monolith.
[0100] The whole is heated to 60 ° C for one hour. The crosslinked gel obtained is washed with hot water until neutral. 50 ml of 2N NaOH and 0.25 g of NaBH 4 are prepared. The monolith is drilled by this solution and is autoclaved at 120 ° C. for one hour.
[0101] The gel is washed with 1N NaOH containing 0.5% hot NaBH4 and then cold. The monolith is rapidly transferred to a pH-buffered ice bath with a solution of acetic acid and sodium acetate. The monolith is washed by circulation of hot distilled water, then iced. EXAMPLE 8 Manufacture of Crosslinked and Functionalized Multicapillary Agarose Pelleting A monolith with porous walls of alpha alumina 150 mm long having 100 conduits 0.45 mm in diameter distributed in a square mesh of 1.2 mm The side is produced by extrusion and sintering of a 20 μm diameter alpha alumina powder. One liter of agarose beads are dissolved in one liter of deionized water at 95 ° C. The porosity of the monolith is impregnated with this solution by soaking and draining the pipes at 95 ° C. and then cooling. The occluded and frozen agarose is washed with distilled water A decilitre of 1N NaOH solution containing 1 ml of epichlorohydrin and 0.25 g of NaBH4 is prepared. We fill the ducts of the monolith. The whole is heated to 60 ° C for one hour. The crosslinked gel obtained is washed with hot water until neutral. 50 ml of 2N NaOH and 0.25 g of NaBH 4 are prepared. The monolith is drilled by this solution and is autoclaved at 120 ° C. for one hour. The gel is washed with 1 N NaOH containing 0.5% warm NaBH4 and then cold. The monolith is rapidly transferred to a pH-buffered ice bath with a solution of acetic acid and sodium acetate. The monolith is washed by circulation of hot distilled water, then iced. The monolith is lyophilized and the dry gel is treated with 60 ml of a mixture of pyridine and acetic acid at equal volumes. Acetylation is conducted by adding 4 ml of acetyl chloride at 60 ° C for 75 minutes, circulating the mixture through the conduits. The monolith is transferred to 100 ml of dry dioxane. 1 g of LiAIH4 are added to the medium, circulating the mixture through the conduits. The temperature is slowly raised to 80 ° C and maintained for 2 hours. The reaction is stopped by adding ethyl acetate and then water.
[0102] The monolith is cooled in an ice bath. 1M hydrochloric acid is circulated through the conduits. The monolith is briefly washed with 0.1M HCl and then with water. Deacetylation is conducted at 80 ° C for 15 minutes with M NaOH containing 0.1% NaBH4. EXAMPLE 9 Manufacture of Multicapillary Polymeric Gel Filling by Immersion of a Hollow Polypropylene Fiber 500 micron-sized polypropylene fibers of CELGARD X30240 polypropylene 300 μm in outer diameter and 240 μm in internal diameter and 300 mm long 10 in the form of a cylinder in a glass tube of 8 mm internal diameter are joined together. A mixture containing 16 g of hydroxyethyl methacrylate, 64 g of divinylbenzene, 890 mg of azobisisobutyronitrile and 120 g of dodecanol was prepared and degassed under nitrogen for 20 minutes. The wall, lumen and interstices of the hollow fiber bundle are impregnated with this mixture by vacuum immersion and then returned to atmospheric pressure followed by drainage of the fiber lumen. The beam is brought to 70 ° C for 24 hours. The mixture polymerizes. The mixture freezes in the porous wall of the fibers under these conditions. The monolith thus produced is washed under THF for 30 minutes and then dried under a stream of dry air at 70 ° C. EXAMPLE 10 Manufacture of multi-capillary polymeric gel packing by molding 2 parts of polypropylene having a MI of 0.8 g / 10 min and one and half part of tetrahydronaphthalene are mixed in a homogeneous mixture at a temperature of about 160 ° C and after cooling to 140 ° C are mixed with one part of styrene containing 8% by weight of dinvinylbenzene. 0.5% by weight of pmethoxyphenol as a polymerization inhibitor and 0.1% by weight of di-t-butyl hydroperoxide are added to the mixture as the polymerization initiator. The mixture is continued for 5 minutes. The mixture is then polymerized in a reactor under pressure at 180 ° C for 8 hours. The resulting polymer is extruded into a sheet 50 mm wide and 0.5 mm thick having 100 grooves 0.4 mm wide and 0.4 mm deep equidistant. The resulting sheet is cut into sheets of lengths 250 mm. The solvent present in these layers is extracted with methanol at boiling point. 100 sheets are then stacked along their length in a square bundle constituting a packing for chromatography. Example 11: Manufacture of multicapillary colloidal silica support pad A polydioxanone wire is produced with a diameter of 0.25 mm.
[0103] The wire is cut into rectilinear sections 15 cm long, covered by dipping with a thin layer of a mixture containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and a volume of Aerosil 380 equivalent to 9/10 of the volume of the solution. The sections are polymerized for 24 hours under nitrogen at 70 ° C. They are then cut into lengths of 120 mm and assembled into a bundle of about 4 mm diameter in a glass tube 100 mm long and 4 mm in diameter previously prepared. A mixture containing 16 g of hydroxyethyl methacrylate, 64 g of divinylbenzene, 890 mg of azobisisobutyronitrile and 120 g of dodecanol is prepared in parallel and degassed under nitrogen for 20 minutes. The previously prepared bundle is filled with this mixture and heated to 70 ° C for 24 hours. The mixture polymerizes. The composite thus produced is released by cutting the sections of wire on either side of the glass tube flush with its ends, perpendicular to said sections.
[0104] The monolith thus produced is washed by percolating THF for 30 minutes and dried in an oven at 70 ° C. The polydioxanone son are dissolved in 90 ° C sodium hydroxide percolated through the packing for 1 hour, then the packing is washed with distilled water until neutral.
[0105] The porosity of the monolith is impregnated with a Ludox SM30 silica sol solution at 30% dry matter by percolating this soil and then draining it out of the lumen of the fibers. The silica sol is dried in situ by circulating dry air at 80 ° C. Example 12 Manufacture of Cellulose Multi-Pillow Lining A monolith with porous walls of alpha alumina 300 mm long having 100 conduits of 0.45 mm in diameter distributed in a square mesh of 1.2 mm side is produced by extrusion and sintering of an alpha alumina powder of 20 μm elemental diameter. 150 g of cellulose for thin layer chromatography (Sigma Aldrich brand) are milled to a particle diameter of 2 μm. The powder obtained is progressively added to 50 ml of 0.35 μm aqueous colloidal polystyrene with a particle size of 5% by weight concentration (PolyScience Inc. mark) until a thick suspension having a viscosity of about 1 Poiseuille is obtained.
[0106] The suspension obtained is poured into the monolith through the ducts so as to fill the porosity of the walls, and the ducts are drained from the suspension they contain. The monolith is dried in an oven at 80 ° C.
[0107] Example 13 Manufacture of a Multicapillary Lining with an Organic Liquid Stationary Phase A mixture is prepared containing 8 g of hydroxyethyl methacrylate, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and degassed under nitrogen during 20 minutes. This mixture is heated at 70 ° C. for 24 hours. The mixture polymerizes. The monolith A thus produced is washed and percolated with THF for 30 minutes and dried in an oven at 90 ° C. Monolith A is milled under liquid nitrogen until a grain diameter of 10 μm is obtained. A polydioxanone wire is produced with a diameter of 0.25 mm. The wire is cut into rectilinear sections 15 cm long, covered by dipping a thin layer of a mixture containing 8 g of styrene, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile, 120 g of dodecanol and a volume of monolith A milled at 10 pm equivalent to 9/10 of the volume of the solution. The sections are polymerized for 24 hours under nitrogen at 70 ° C. They are then cut into lengths of 120 mm and assembled into a bundle of about 4 mm diameter in a glass tube 100 mm long and 4 mm in diameter previously prepared. A mixture containing 8 g of styrene, 32 g of divinylbenzene, 445 mg of azobisisobutyronitrile and 120 g of dodecanol is prepared in parallel and degassed under nitrogen for 20 minutes. The previously prepared bundle is filled with this mixture and heated to 70 ° C for 24 hours. The mixture polymerizes. The composite thus produced is released by cutting the sections of wire on either side of the glass tube flush with its ends, perpendicular to said sections. The monolith thus produced is washed by percolating THF for 30 minutes and dried in an oven at 90 ° C. The polydioxanone son are dissolved with N sodium hydroxide at 90 ° C containing anionic surfactant and percolated through the packing for 1 hour, then the packing is washed with distilled water until neutral. The lining is impregnated by immersion in cyclohexyl acetate followed by draining of the lumen of the conduits. EXAMPLE 14 Manufacture of a Multicapillary Lining by Immersion of a Hollow Fiber of Polypropylene in an Organic Liquid
[0108] 500 micron-sized polypropylene fibers of CELGARD X30-240 polypropylene 300 μm in outer diameter and 240 μm in internal diameter and 300 mm long in the form of a cylinder in a glass tube having an internal diameter of 8 mm are aligned.
[0109] The wall, lumen and interstices of the hollow fiber bundle between the fibers are impregnated with cyclohexyl acetate by total immersion in vacuo and then returned to atmospheric pressure followed by drainage of the fiber lumen. The interstices between the individual fibers are filled with cyclohexyl acetate Example 15: Manufacture of a multicapillary packing with an organic liquid stationary phase A monolith with porous walls of 300 mm long pyrex glass having 100 conduits of 0.45 mm Diameter distributed in a square mesh of 1.2 mm side is produced by extrusion and sintering of a pyrex glass powder of 20 pm of elementary diameter. The lining is immersed in hexamethyl disilazane and heated to 140 ° C under pressure in a closed container so as to hydrophobise the surface of the glass. The lining is impregnated by immersion in cyclohexyl acetate followed by drainage of the lumen of the ducts.
[0110] EXAMPLE 16 Manufacture of Multicapillary Sepharose Lining A monolith with porous walls of alpha alumina of 300 mm in length and having 100 conduits of 0.45 mm in diameter distributed in a square mesh of 1.2 mm on one side is produced by extrusion and sintering of an alpha alumina powder of 20 μm in elementary diameter. 150 g of Sepharose (GE HealthCare) are ground in liquid nitrogen to a particle diameter of 2 μm. The powder obtained is progressively added to 50 ml of aqueous colloidal polystyrene of 0.80 .mu.m of particle size at 5% concentration by weight (PolyScience Inc. mark) until a thick suspension having a viscosity of about 1 Poiseuille is obtained.
[0111] The suspension obtained is poured into the monolith through the ducts so as to fill the porosity of the walls, and the ducts are drained from the suspension they contain. The monolith is dried in an oven at 80 ° C.
[0112] REFERENCES [1] "VVater Diffusion and Permeability in Unsaturated Polyester Resins Flows Characterized by Measurement Performed with a VVater-Specific Permeate: Analysis of the Transient Permeation", S. Marais, M. Métayer, M. Labbé, Journal of Polymer Science, Dec. 1999, Vol. 74, Issue 14, pp. 3380-3396 [2] Chromatographies in liquid and supercritical phases, R. Rosset, M. Caudé, A. Jardy, MASSON 3rd edition, 1991 [3] Measurement of the Effective Diffusivity in Porous Media by the diffusion Oeil Method. Soo Park, Duong D. Do, Catalysis Review: Science and Engineering, 1996, Vol. 38, Issue 2 pp. 189-247

权利要求:
Claims (32)
[0001]
REVENDICATIONS1. A chromatography process in which a gaseous, liquid or supercritical mobile phase containing species to be separated is circulated through a lining, said lining comprising: a plurality of capillary ducts passing through the lining between a so-called upstream face through which the phase mobile penetrates the lining and a so-called downstream face through which the mobile phase leaves the lining, and - a continuous medium permeable to molecular diffusion extending between said ducts, comprising a porous organic gel or an organic liquid and including at least one a network of related pores whose size is greater than twice the molecular diameter of the species to be separated and open on the ducts, so as to provide said species with a diffusive path between said ducts.
[0002]
2. Method according to claim 1, wherein the average molar diffusion rate of the species to be separated between the adjacent ducts under the effect of a given concentration difference of said species between the walls of said ducts is greater than 0.01 times the molar flow rate. means of diffusion of the species a conduit and the stationary phase that constitutes the packing under the effect of the same difference in concentration of the species to be separated between the fluid led by the conduits and the wall of said conduits.
[0003]
3. Method according to one of claims 1 or 2, wherein the permittivity of said continuous medium vis-à-vis the species to be separated is greater than 5000 Barrer, that is to say greater than 5 e-7 (cm3 02 cm) / (cm 2 cm Hg).
[0004]
4. Method according to one of claims 1 to 3, wherein the diameter of the capillary ducts of the lining is less than or equal to 500 pm, preferably less than or equal to 150 pm and more preferably less than or equal to 50 pm.
[0005]
5. Method according to one of claims 1 to 4, wherein said continuous medium is formed of an organic gel, said organic gel being selected from: (a) a copolymer of styrene and divinylbenzene, (b) polymethacrylate of methyl, (c) a copolymer of hydroxyethyl methacrylate and divinylbenzene.
[0006]
6. Method according to one of claims 1 to 4, wherein said continuous medium is formed of an organic gel, said organic gel being a polysaccharide.
[0007]
7. The method according to one of claims 1 to 4, wherein said continuous medium is formed of an organic liquid extending in said network of related pores, said organic liquid being selected from: (a) an aliphatic or aromatic hydocarbon (b) an aliphatic or aromatic alcohol, (c) an aliphatic or aromatic ketone, (d) an aliphatic or aromatic amine, (d) a halogenated organic compound.
[0008]
The method according to one of claims 1 to 6, wherein the packing comprises a molecular diffusion permeable organic gel monolith through which said capillary conduits extend, said network of related pores extending within said gel. organic.
[0009]
The method according to one of claims 1 to 7, wherein the packing comprises a monolith of a chemically inert porous material containing said network of related pores, said pores being filled with said organic gel or said molecular diffusion-permeable organic liquid. .
[0010]
The method according to one of claims 1 to 9, wherein the packing comprises a monolith of a chemically inert porous material containing said continuous network of pores, the surface of said pores being coated with the organic permeable gel to molecular diffusion on a thickness chosen so as to leave, in said pore network, a free volume for the diffusion of the mobile phase, said organic gel forming a continuous network of pores between the conduits.
[0011]
11. Method according to one of claims 9 or 10, wherein the chemically inert material of said monolith is selected from silica, alumina, or a combination of silica and alumina.
[0012]
12. Method according to one of claims 1 to 7, wherein the lining comprises a stack of porous fibers each comprising a lumen forming a capillary duct of the lining and a wall comprising a network of related pores, said fibers being made joined by the porous organic gel or organic liquid permeable to molecular diffusion.
[0013]
The method of claim 12, wherein the wall of each fiber is formed from said organic permeable molecular diffusion gel.
[0014]
The method of claim 12, wherein the pores of the wall of each fiber are filled with said gel or said organic liquid permeable to molecular diffusion.
[0015]
15. The method of claim 12, wherein the pore surface of the wall of each fiber is covered with the organic permeable gel molecular diffusion to a selected thickness so as to leave, in said pore network, a free volume for the diffusion of the mobile phase, said organic gel forming a continuous network of pores inside said wall.
[0016]
16. The method as claimed in one of claims 1 to 15, in which the organic permeable gel for molecular diffusion forms the chromatographic stationary phase.
[0017]
17. Method according to one of claims 1 to 15, wherein the organic gel has pores containing a solid third body forming the chromatographic stationary phase.
[0018]
18. A method of manufacturing a packing for the implementation of the chromatography method according to one of claims 8 to 11, comprising the following steps: - providing a son beam called precursors of the capillary ducts, - formation of a porous matrix around the wires or ducts, so as to form a monolith, - removing the wires so as to form said capillary ducts.
[0019]
19. The method of claim 18, wherein the matrix is an organic gel.
[0020]
The method of claim 18, wherein the matrix comprises a chemically inert material and is loaded with said matrix of an organic gel. 35
[0021]
21. Manufacturing method according to one of claims 19 to 20, wherein the precursor son of the capillary ducts are fusible son at a temperature below the degradation temperature of the matrix and the elimination of said son comprises the fusion and the draining said wires out of the lining.
[0022]
22. The method of claim 21, wherein the fusible wires comprise indium, bismuth, tin, gallium, silver or an alloy thereof with other metals other than lead. , mercury and cadmium.
[0023]
23. A method of manufacturing a packing for the implementation of the chromatography method according to one of claims 12, 14 or 15, comprising the steps of: - providing a compact bundle of hollow fibers, - inclusion in the porous wall of the hollow fibers of an organic gel or a precursor of said organic gel to be polymerized in situ, so as to leave the lumen of the hollow fibers free and open, - creation of a diffusive bonding between said hollow fibers by said gel or organic liquid.
[0024]
24. A method of manufacturing a packing for carrying out the chromatography method according to one of claims 1 to 6, wherein is carried a molding of the organic gel in a structure defining said capillary ducts.
[0025]
25. Padding for chromatography, comprising: a plurality of capillary ducts passing through the packing between a so-called upstream face intended for the entry of the phase into the packing and a so-called downstream face intended for the exit of the mobile phase of the packing, and a continuous molecular diffusion permeable medium extending between said conduits, comprising a porous organic gel or an organic liquid and including at least one family of related pores.
[0026]
26. Padding according to claim 25, wherein the diameter of the capillary ducts of the packing is less than or equal to 500 μm, preferably less than or equal to 150 μm and more preferably less than or equal to 80 μm.
[0027]
27. Padding according to one of claims 25 or 26, wherein said continuous medium is formed of an organic gel, said organic gel being selected from: (a) a copolymer of styrene and divinylbenzene, (b) polymethacrylate of methyl, (c) a copolymer of hydroxyethyl methacrylate and divinylbenzene.
[0028]
28. Padding according to one of claims 25 or 26, wherein said continuous medium is formed of an organic gel, said organic gel being a polysaccharide.
[0029]
29. The liner as claimed in claim 25, in which said continuous medium is formed of an organic liquid extending in the network of related pores, said organic liquid being chosen from: (a) an aliphatic or aromatic hydocarbon (b) an aliphatic or aromatic alcohol, (c) an aliphatic or aromatic ketone, (d) an aliphatic or aromatic amine, (d) a halogenated organic compound.
[0030]
30. Padding according to one of claims 25 to 28, comprising a molecular diffusion-permeable organic gel monolith through which said capillary ducts extend.
[0031]
31. The liner according to one of claims 25 to 29, comprising a monolith of a porous chemically inert material having a continuous network of pores, said pores being filled with said gel or said organic liquid permeable to molecular diffusion.
[0032]
32. Padding according to claim 31, comprising a monolith of a porous chemically inert material having a continuous network of pores, the surface of said pores being covered with the organic permeable gel molecular diffusion to a thickness selected so as to retain, in said pore network, a free volume for the diffusion of the mobile phase, said organic gel forming a continuous network of pores between the conduits.

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引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

US20070017870A1|2003-09-30|2007-01-25|Belov Yuri P|Multicapillary device for sample preparation|

---

WO2011114017A2|2010-03-15|2011-09-22|Parmentier Francois|Multicapillary monolith|

---

WO2013064754A1|2011-09-15|2013-05-10|Parmentier Francois|Multi-capillary monolith made from amorphous silica and/or activated alumina|FR3049874A1|2016-04-06|2017-10-13|Francois Parmentier|CHROMATOGRAPHY PROCESS|US4007138A|1972-05-25|1977-02-08|Badische Anilin- & Soda-Fabrik Aktiengesellschaft|Manufacture of ion-exchanging shaped articles|

---

US4957620A|1988-11-15|1990-09-18|Hoechst Celanese Corporation|Liquid chromatography using microporous hollow fibers|

---

US6749749B2|2002-06-26|2004-06-15|Isco, Inc.|Separation system, components of a separation system and methods of making and using them|

---

US7473367B2|2002-06-26|2009-01-06|Dionex Corporation|Monolithic column|

---

JP4314219B2|2005-07-04|2009-08-12|株式会社東芝|Filter circuit and wireless communication apparatus using the same|

---

US8017015B2|2006-10-20|2011-09-13|Quest Diagnostics Investments Incorporated|Monolithic column chromatography|

---

JP5290603B2|2007-05-28|2013-09-18|オルガノ株式会社|Particle aggregation type monolithic organic porous body, method for producing the same, particle aggregation type monolithic organic porous ion exchanger, and chemical filter|

---

WO2009020649A1|2007-08-08|2009-02-12|Jordi Flp|Suspension homopolymerization of an isocyanurate|

---

JP5525848B2|2009-03-18|2014-06-18|オルガノ株式会社|Ion chromatography device column, suppressor and ion chromatography device|

---

FR3026313B1|2014-09-29|2018-07-13|Francois Parmentier|METHOD OF CHROMATOGRAPHY ON A MULTICAPILLARY TRIM|FR3041547B1|2015-09-29|2019-09-20|Francois Parmentier|METHOD OF CHROMATOGRAPHY ON A POROUS TRIM MADE BY STRETCHING|

---

CN108951249B|2018-06-22|2020-11-24|洛阳师范学院|Palm single fiber microtubule, capillary gas chromatographic column, preparation and application thereof|

---

FR3112083A1|2020-07-03|2022-01-07|François PARMENTIER|Manufacturing process of a multicapillary filling|

---

GB202010885D0|2020-07-15|2020-08-26|Johnson Matthey Plc|Methods for the separation and/or purification of metals|

法律状态:
2015-09-25| PLFP| Fee payment|Year of fee payment: 2 |

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2016-04-01| PLSC| Publication of the preliminary search report|Effective date: 20160401 |

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2016-09-30| PLFP| Fee payment|Year of fee payment: 3 |

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2017-09-26| PLFP| Fee payment|Year of fee payment: 4 |

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2019-09-26| PLFP| Fee payment|Year of fee payment: 6 |

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2021-02-24| PLFP| Fee payment|Year of fee payment: 7 |

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2021-09-30| PLFP| Fee payment|Year of fee payment: 8 |

优先权:

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申请号 | 申请日 | 专利标题

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FR1459175A|FR3026312B1|2014-09-29|2014-09-29|PROCESS FOR CHROMATOGRAPHY ON A GEL OR ORGANIC LIQUID|FR1459175A| FR3026312B1|2014-09-29|2014-09-29|PROCESS FOR CHROMATOGRAPHY ON A GEL OR ORGANIC LIQUID|

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US15/515,082| US10864461B2|2014-09-29|2015-09-29|Organic gel or liquid chromatography method|

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JP2017535965A| JP6858701B2|2014-09-29|2015-09-29|Organic gel or liquid chromatography method|

---

EP15771943.6A| EP3200890A1|2014-09-29|2015-09-29|Organic gel or liquid chromatography method|

---

CN201580063426.4A| CN107107029A|2014-09-29|2015-09-29|Organogel or liquid chromatography|

---

PCT/EP2015/072466| WO2016050789A1|2014-09-29|2015-09-29|Organic gel or liquid chromatography method|

---

US17/096,456| US20210069613A1|2014-09-29|2020-11-12|Organic gel or liquid chromatography method|