WO2015067912A2 - Procédé de stockage et de concentration d'un composé non volatil - Google Patents
Procédé de stockage et de concentration d'un composé non volatil Download PDFInfo
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- WO2015067912A2 WO2015067912A2 PCT/FR2014/052866 FR2014052866W WO2015067912A2 WO 2015067912 A2 WO2015067912 A2 WO 2015067912A2 FR 2014052866 W FR2014052866 W FR 2014052866W WO 2015067912 A2 WO2015067912 A2 WO 2015067912A2
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- WIPO (PCT)
- Prior art keywords
- porous substrate
- volatile compound
- microchannel
- compound
- fluid
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/405—Concentrating samples by adsorption or absorption
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5023—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures with a sample being transported to, and subsequently stored in an absorbent for analysis
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0678—Facilitating or initiating evaporation
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/069—Absorbents; Gels to retain a fluid
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
Definitions
- the present invention is directed to a method of storing and concentrating a nonvolatile compound. It applies, in particular, to medical diagnostic systems whose samples are in small quantities.
- microfluidic system refers to a system handling fluids of which at least one of the characteristic dimensions is between 1 micrometer and 500 micrometers. These systems have the advantage of requiring a very small amount of an analyte to function.
- a “reagent” is any compound that may interact with an analyte. This interaction may be of a chemical nature, for example with proton exchange, electron exchange, formation and / or rupture of chemical bonds of the covalent bond type, hydrogen bonds, disulphide bridges or Van der Waals bonds. This interaction can also be electrostatic, repulsive or attractive. This interaction may be specific, for example with the formation of antigen-antibody complexes, the formation of enzyme-substrate complexes, the hybridization of complementary DNA strands, or nonspecific DNA strands.
- volatile compound refers to any compound in which a large part of the volume evaporates during the experimental times considered.
- non-volatile compound refers to any compound whose volume which evaporates during the considered experiment time is negligible.
- Microfluidic systems are increasingly used in areas as diverse as chemistry, biology, physics, analysis, screening. There are different types of these systems, including chips from the microfabrication of glass, silicon, metal, polymers or a combination of these materials.
- microchannels can be etched in the substrate by any known method. A solid or thin layer then covers the substrate, thus defining the geometry of the microchannels.
- the microchannels can also be obtained by molding an elastomer in a suitable mold and then placed on a substrate. These microchannels can be arranged to form a network in which fluids circulate.
- the flows are generated, most of the time, by external energy sources such as pumps for pressure control and syringe pumps acting on the flow.
- external energy sources such as pumps for pressure control and syringe pumps acting on the flow.
- a more autonomous microfluidic is obtained by exploiting the capillary forces and the wetting properties.
- US Pat. No. 7,695,687 illustrates the autonomous microfluidics in which the capillary forces make it possible to generate a flow.
- a porous substrate which has the advantage of naturally presenting a network of microchannels. This is, for example, the case of paper in which the water flows spontaneously.
- microfluidic paper systems are particularly applicable in the field of medical diagnostics because these technologies can be deployed on a large scale and at low cost.
- a drop of fluid comprising at least one non-volatile compound is deposited on a paper containing a reagent configured to react with at least one non-volatile compound.
- the medical tests performed with these systems give almost instantaneous results and are for single use only.
- microfluidic paper systems do not optimize the amount of nonvolatile compound, for example present in a sample, necessary to carry out a reaction between a reagent and at least one nonvolatile compound.
- current systems store samples in liquid form in microchannels or containers.
- document FR 2 946 269 which relates to a microfluidic device for conveying a product from a product injection zone to a product arrival zone through a product channeling zone. Because of this conveying objective, the device aims to minimize the evaporation effect of a solvent transporting the product during the conveyance.
- This document is not intended to use a dried sample present on a porous substrate but aims at a technical education of a transport solution of a sample.
- Patent application EP 2,560,004 is also known which relates to a device for detecting the presence of a compound in a fluid.
- the technical effect sought is to cause a chemical reaction of a compound attached to a porous substrate in the presence of the compound to be detected so as to color the porous substrate for example.
- This document does not teach a non-volatile compound transport mechanism for satisfactory storage and retrieval in view of the constraints mentioned above.
- patent application WO 00/54309 is known which relates to a mass spectrometry device for determining the mass of a target molecule to determine the nature of this molecule. This document does not allow optimized storage of the molecules in and / or on a porous substrate with regard to the constraints mentioned above.
- the present invention aims to remedy all or part of these disadvantages.
- the present invention aims, in a first aspect, a method of storage and concentration of at least one non-volatile compound present in a fluid further comprising at least one volatile compound, which comprises:
- a concentration step of each non-volatile compound transported comprising:
- a step of drying each non-volatile compound at a location of the porous substrate made by evaporation of each volatile compound for each step of injecting a volatile compound, a step of drying each non-volatile compound at a location of the porous substrate made by evaporation of each volatile compound.
- the method which is the subject of the present invention comprises, downstream from the concentration step, a step of reaction of at least one non-volatile compound with at least one reagent, each said reagent being configured to react with each said non-volatile compound.
- the method of the present invention comprises, upstream of the concentration step, a step of separating each non-volatile compound at a different location from the porous substrate by differentiated displacement of each non-volatile compound.
- the reaction step employs a plurality of reagents, each reagent being positioned at a different location from the porous substrate so that at least two reagents do not come into contact with each other.
- reagents staining in contact with a non-volatile compound to separately identify the presence of at least two non-volatile compounds.
- the reagent is configured to, during a reaction step, modify the transport properties of the nonvolatile compound.
- the method which is the subject of the present invention comprises a plurality of additional steps for injecting a fluid into or onto the porous substrate so as to concentrate at the same place in the porous substrate, successively, different quantities of the same non-volatile compound.
- the method that is the subject of the present invention comprises a step of destocking at least one non-volatile compound from the porous substrate to a container, by passing through the porous substrate by a solvent in which said non-volatile compound dissolves .
- At least a portion of the porous substrate has a fibrous density different from the rest of the porous substrate.
- the porous substrate has a fiber density gradient along an axis of the porous substrate.
- the porous substrate has a barrier on at least a portion of the surface of the porous substrate, the barrier being configured to be non-porous for at least one non-volatile compound, the porous substrate being configured to be porous, for at least one volatile compound, including under the barrier.
- the present invention aims, according to a second aspect, a storage and concentration support of at least one non-volatile compound present in a fluid further comprising at least one volatile compound, which comprises:
- a porous substrate for at least one volatile compound, including under a barrier and the barrier on at least a portion of the surface of the porous substrate, the barrier being configured to be non-porous for at least one non-volatile compound, at least one non-volatile compound is thus concentrated by flow in the porous substrate of at least one volatile compound.
- the non-volatile compound is spatially concentrated on the porous substrate according to the positioning of the barrier.
- the flow of each volatile compound in the entire volume of the porous substrate allows a rapid concentration of each nonvolatile compound.
- the barrier is positioned at least partially in the porous substrate.
- the barrier is obtained by solidifying a polymer or resin on the surface of the porous substrate.
- the barrier is obtained by melting and then solidifying wax on the porous substrate.
- the barrier is a non-porous adhesive tape for at least one non-volatile compound.
- the advantage of these embodiments is that they allow a low cost installation of a surface barrier or a partial thickness of the porous substrate.
- the barrier is obtained by local contrast of wetting property of the porous substrate.
- the barrier forms a transport channel of each nonvolatile compound.
- the transport channel is configured:
- each non-volatile compound is transported, at least by capillarity, by at least one volatile compound along the channel and
- each non-volatile compound is concentrated in different places of the porous substrate by evaporation of each volatile compound.
- the at least one porous substrate comprises a reagent configured to react with at least one nonvolatile compound.
- a reagent configured to react with at least one nonvolatile compound.
- the porous substrate has a plurality of reagents such that at least two reactive compounds do not come into contact with each other. The advantage of these embodiments is that, in the case of reagents configured to adopt a certain color in case of detection of a certain non-volatile compound, the two colors do not overlap so as to allow easy identification of the presence of each non-volatile compound reacted.
- the barrier is configured to allow attachment of at least one other porous substrate to contact at least two porous substrates.
- the advantage of these embodiments is that they allow the transport of a non-volatile compound from one porous substrate to another.
- At least one other porous substrate attached to the support has a fibrous density different from the fibrous density of the porous substrate.
- the barrier is configured to increase in thickness along an axis of the porous substrate so as to achieve the cross-concentration of each nonvolatile compound in the porous substrate.
- the present invention relates to a device comprising:
- a means for injecting a fluid on the porous substrate configured to perform a plurality of injections so as to concentrate at the same location of the porous substrate, successively, different amounts of the same non-volatile compound.
- a non-volatile compound can be concentrated in the same place of the porous substrate.
- this aspect makes it possible, with a single injection of non-volatile compound, to increase the concentration at a location of the porous substrate by successive injection of volatile compound.
- the present invention is directed to a device comprising:
- a means for injecting a fluid onto the porous substrate configured to produce a plurality of injections of different fluids, at least one fluid of which comprises at least one non-volatile compound on a porous substrate so as to concentrate in different locations of the substrate porous, various nonvolatile compounds or so as to displace at least one nonvolatile compound from one place to another of the porous substrate.
- the device that is the subject of the present invention comprises a means for transporting at least one non-volatile compound from the porous substrate to a container, by passing through the porous substrate by a solvent in which the non-volatile compound dissolves .
- the device which is the subject of the present invention comprises a means of selective destocking of a non-volatile compound by passing through the place in which the non-volatile compound is concentrated by a solvent in which the compound dissolves.
- the present invention provides a method of transporting and retrieving at least one non-volatile compound dried in a porous substrate, which comprises:
- a destocking step of each non-volatile compound in at least one microchannel which comprises:
- the present invention provides a method for transporting and storing at least one non-volatile compound present in a fluid that comprises:
- a step of storing each nonvolatile compound in a porous substrate which comprises:
- each non-volatile compound in the porous substrate makes it possible to store, on a porous substrate, at least one nonvolatile compound present in a microchannel in dried form. This dried nonvolatile compound can then be destocked for use in other applications.
- the transport step comprises a step of joining the porous substrate with at least one microchannel.
- the method that is the subject of the present invention comprises, upstream of the transport step, a step of opening at least one microchannel so as to allow the insertion of a part of the porous substrate into said microchannel.
- the method that is the subject of the present invention comprises, upstream of the injection step, a step of insertion of the porous substrate into at least one microchannel.
- the method that is the subject of the present invention comprises, upstream of the drying step, a step of spatial concentration of each non-volatile compound in the porous substrate.
- the spatial concentration step of each non-volatile compound is performed at different locations of the porous substrate.
- the fluid further comprises a volatile compound configured to allow the transportation of each nonvolatile compound and to evaporate during the drying step.
- a volatile compound configured to allow the transportation of each nonvolatile compound and to evaporate during the drying step.
- the method which is the subject of the present invention comprises a step of selective destocking of a non-volatile compound by passing through the place in which the non-volatile compound is concentrated by a solvent in which the compound dissolves.
- the destocking step comprises a step of dividing the porous substrate into zones in which at least one nonvolatile compound is concentrated.
- the advantage of these embodiments is to improve the achievement of a selective destocking of at least one nonvolatile compound by avoiding pollution to other non-volatile compounds.
- the present invention relates to a method for transporting, storing and retrieving at least one non-volatile compound present in a fluid, which comprises:
- a step of storing each nonvolatile compound in a porous substrate which comprises:
- a destocking step of each non-volatile compound in at least one microchannel which comprises:
- FIG. 1 represents, in the form of a logic diagram, steps of a particular embodiment of the storage and retrieval method of the present invention
- FIG. 2 represents, in the form of a logic diagram, steps of a particular embodiment of the storage and concentration method that is the subject of the present invention
- FIG. 3 represents, schematically, a particular embodiment of a storage and concentration medium which is the subject of the present invention
- FIG. 4 represents, schematically, a particular embodiment of a storage and concentration device which is the subject of the present invention
- FIG. 5 represents, schematically, a particular embodiment of a storage and concentration device which is the subject of the present invention
- FIGS. 6 to 8 show particular embodiments of a microchannel whose number of contact zones and injection-recovery zones varies
- FIGS. 9 to 11 represent a particular embodiment of a microchannel implemented, for example, by the method represented in FIG. 1,
- FIG. 12 represents a particular embodiment of a manual system for regulating the internal pressure of a microchannel
- FIGS. 13 and 14 show two particular modes of opening of a microchannel and bringing into contact a porous substrate
- FIGS. 15 and 16 show particular embodiments of a microchannel
- FIG. 17 represents a particular embodiment of a transfer of liquid from a porous substrate to a microchannel
- FIG. 18 represents a microchannel in which a porous substrate has been inserted
- FIGS. 19 to 22 represent the displacement of a non-volatile compound through a porous substrate according to the characteristics of the non-volatile compound
- FIGS. 23 and 24 represent, by two curves, the evolution of the concentration of a non-volatile compound on a porous substrate as a function of parameters of particular injections,
- FIGS. 25 to 30 show effects, in terms of spatial concentration, of different types of barriers of a porous substrate
- FIGS. 31 to 34 represent different types of barriers
- FIG. 35 shows a microchannel made from barriers placed on a porous substrate.
- porous substrates implemented in the embodiments below are formed, for example, by a network of fibers of characteristic diameter of 20 microns, forming polydispersed pores that can have a mean transverse dimension of five to ten micrometers.
- porous substrate is meant, for example, a paper sheet having a thickness of 200 microns and a centimeter width.
- FIG. 1 shows a particular embodiment of the method 100 of transport, storage and retrieval.
- This method 100 comprises: a step 105 for opening at least one microchannel so as to allow the insertion of a portion of a porous substrate into each said microchannel,
- a step 165 of destocking each non-volatile compound in at least one microchannel which comprises:
- the opening step 105 of at least one microchannel so as to allow the insertion of a portion of the porous substrate in said microchannel is carried out, for example, by the implementation of a section means.
- This section means may be, for example, a pair of scissors.
- This opening step 105 notably makes it possible to avoid too rapid evaporation of a volatile compound contained in the microchannel.
- a particular embodiment of this opening step 105 is detailed, below, with reference to FIG.
- the step of bringing the porous substrate into contact with at least one microchannel is carried out, for example, by inserting the porous substrate into an opening of a microchannel.
- the intended application consists mainly of the storage and destocking of a non-volatile compound.
- the advantage of inserting a porous substrate into a microchannel, having two openings for example, is to allow the non-volatile compounds entering the microchannel to be concentrated on the porous substrate. In this way, the recovery of non-volatile compounds is facilitated since the non-volatile compounds are concentrated.
- a particular embodiment of a device corresponding to this contacting step 1 10 is described below, with reference to FIG. 18.
- the contacting step 1 10 is carried out, for example, by juxtaposition of an opening of at least one microchannel and the porous substrate.
- the step 1 of joining the porous substrate with at least one microchannel is carried out, for example, by the implementation of fixing clips configured to press the porous substrate against an open microchannel.
- a particular embodiment of this joining step 1 is detailed below, with reference to FIG.
- the fluid injection step 125 in at least one microchannel formed in a non-porous material for each non-volatile compound is carried out, for example, by the implementation of a syringe depositing in an opening of each said microchannel fluid comprising at least one non-volatile compound.
- this method 100 comprises a plurality of fluid injection steps 125 in at least one microchannel, so as to concentrate at the same location of the porous substrate, successively, different amounts of the same non-volatile compound.
- this method 100 comprises a plurality of different fluid injection stages 125, at least one fluid of which comprises at least one non-volatile compound in at least one microchannel, so as to concentrate the porous substrate in different places. , various nonvolatile compounds or so as to displace at least one nonvolatile compound from one place to another of the porous substrate.
- the additional injection of a fluid comprising a non-volatile compound present during a previous injection step 125 makes it possible to increase the concentration of this non-volatile compound in the porous substrate.
- the injection 125 of a fluid comprising at least one volatile compound, for example a solvent makes it possible to displace at least one nonvolatile compound.
- This effect may be particularly interesting in the context of a porous substrate comprising a reagent configured to react with a particular non-volatile compound.
- the effects of these additional injections are described below in the descriptions of Figures 20-24.
- the modification step 130 of the pressure of the fluid injected into at least one microchannel so as to cause the flow of the fluid from the microchannel to the porous substrate or a fluid from the porous substrate to the microchannel is carried out, for example, by the implementation of a syringe-type syringe.
- By increasing the pressure in the microchannel the fluid is pushed from the microchannel to the porous substrate.
- By lowering the pressure in the microchannel the fluid is drawn from the porous substrate to the microchannel.
- a device for performing this step 130 of modifying the pressure is detailed below, in the description of FIG. 12.
- the step 135 for transporting each non-volatile compound in the porous substrate for each non-volatile compound is carried out by capillarity.
- the spatial concentration step is carried out by evaporation of each volatile compound carrying at least one non-volatile compound.
- concentration of at least one nonvolatile compound at a site of the porous substrate can be improved by additional injection of volatile compound allowing the transport of portions of the nonvolatile compound that have not moved sufficiently before drying of the volatile compound.
- spatial concentration of each non-volatile compound is produced at different locations of the porous substrate by evaporation of each non-volatile compound.
- the division step 150 of the porous substrate in zones in which at least one non-volatile compound is concentrated is carried out, for example, by section of the porous substrate in different zones.
- a barrier traversing the porous substrate in its thickness and in its width makes it possible to divide the porous substrate into zones.
- the contacting step 1 10 is carried out, for example, by juxtaposing an opening of at least one microchannel and the porous substrate.
- the step 1 of joining the porous substrate with at least one microchannel is carried out, for example, by the implementation of fixing clips configured to press the porous substrate against an open microchannel.
- the step 170 of injecting a fluid onto or into the porous substrate is carried out, for example, by the implementation of a syringe depositing a fluid comprising at least one non-volatile compound on or in the porous substrate.
- the selective destocking step of a nonvolatile compound is conducted through a zone, which includes a locus in which a nonvolatile compound is concentrated, by a solvent in which the compound dissolves.
- the modification step 130 of the pressure of the fluid injected into at least one microchannel so as to cause the flow of the fluid from the porous substrate to the microchannel is carried out, for example, by the implementation of a syringe of the type syringe driver. By reducing the pressure in the microchannel, the fluid is drawn from the porous substrate to the microchannel.
- the step 135 of transporting at least one nonvolatile compound from the porous substrate to at least one microchannel is carried out by passing through the porous substrate by a solvent in which said nonvolatile compound dissolves. This transport is carried out, for example, by capillarity.
- FIG. 2 shows a particular embodiment of the storage and concentration method 200.
- This method 200 comprises:
- a step 215 of concentration of each non-volatile compound transported comprising:
- a step 230 of reacting at least one non-volatile compound with at least one reagent each said reagent being configured to react with each said non-volatile compound
- the fluid injection step 205 in at least one microchannel formed in a non-porous material for each non-volatile compound is carried out, for example, by the implementation of a syringe depositing in or on a porous substrate a fluid comprising at least one non-volatile compound.
- the transport step 210 of the fluid in or on the porous substrate is carried out, for example, by capillarity.
- at least one volatile compound carries at least one nonvolatile compound by capillarity through the porous substrate.
- the porous substrate comprises a fiber density gradient along a longitudinal axis of flow so as to gradually retain the non-volatile compounds as a function of the size of these non-volatile compounds.
- the surface of the porous substrate includes a barrier configured to be non-porous for at least one non-volatile compound, the porous substrate being configured to be porous for at least one volatile compound, including under the barrier.
- the volatile compound is dissipated in the thickness of the porous substrate so as to increase the volume and evaporation surface of the volatile compound. Conversely, the non-volatile compound is retained on the surface and moves only on the surface of the porous substrate.
- the separation step 235 is performed, for example, by the difference in flow rate of at least two non-volatile compounds.
- each non-volatile compound has its own characteristics which influence the speed of displacement of this compound non-volatile in the porous substrate. This displacement, moreover, is limited by the time necessary for the evaporation of each volatile compound in which the nonvolatile compound is dissolved. Thus, each nonvolatile compound moves to a certain place of the porous substrate for the time necessary for evaporation of the solvent. If two non-volatile compounds have a different rate of displacement, the two non-volatile compounds separate during this separation step 235.
- the additional injection step 220 is performed, for example, by the implementation of a syringe depositing in or on a porous substrate a fluid comprising at least one volatile compound. Each additional injection step 220 displaces at least one nonvolatile compound dissolving in at least one volatile compound before being transported by each volatile compound.
- the additional injection step 240 is performed, for example, by the implementation of a syringe depositing in or on a porous substrate a fluid comprising at least one non-volatile compound.
- Each additional injection step 240 makes it possible to increase the amount of at least one nonvolatile compound in the porous substrate, each nonvolatile compound thus injected can then be concentrated in a place of the porous substrate.
- the drying step 225 is carried out, for example, by evaporation of each volatile compound present on or in the porous substrate.
- the reaction step 230 is performed, for example, by the deposition of a reagent at a location of the porous substrate.
- This reagent is configured to react with at least one non-volatile compound.
- a non-volatile compound is transported to the place where the reagent is placed, the reagent and the non-volatile compound react.
- a plurality of reagents are positioned at different locations of the porous substrate.
- At least one reagent is configured to modify the displacement properties of at least one nonvolatile compound such as, for example, to attach a nonvolatile compound to a reagent.
- the destocking step 245 is carried out by passing through a place, which comprises a concentrated nonvolatile compound, with a solvent in which the compound dissolves.
- the solvent is then, for example, sucked by a microchannel whose pressure is reduced so as to cause a flow of the solvent from the porous substrate to the microchannel.
- FIG. 3 shows a particular embodiment of the storage and concentration support 300 which is the subject of the present invention.
- This support 300 comprises:
- a porous substrate 305 for at least one volatile compound, including under a barrier 310, which comprises a plurality of reagents 320 configured to react with at least one non-volatile compound and
- the barrier 310 on at least a portion of the surface of the porous substrate, the barrier being configured to be non-porous for at least one non-volatile compound, at least one non-volatile compound is thus concentrated by flow in the porous substrate of at least one volatile compound.
- the porous substrate 305 is, for example, a sheet of paper on which a barrier 310 is positioned.
- This barrier 310 is, for example, formed with wax deposited on the porous substrate 305 and then melted so as to penetrate into the porous substrate 305.
- This barrier 310 forms, on and in the porous substrate 305, a transport channel 315 for at least one volatile compound comprising at least one non-volatile compound.
- the transport channel 315 is configured:
- each non-volatile compound is transported, at least by capillarity, by at least one volatile compound along the channel 315 and
- each non-volatile compound is concentrated in different locations of the porous substrate 305 by evaporation of each volatile compound.
- Each nonvolatile compound exhibits different porous substrate displacement properties 305 depending on characteristics specific to each nonvolatile compound.
- the differences in the rate of displacement on the porous substrate 305 of each nonvolatile compound cause a concentration in different places of each nonvolatile compound according to the time required for the evaporation of each volatile compound.
- the barrier 310 is configured to allow the attachment of at least one other porous substrate, not shown, so as to contact at least two porous substrates. This barrier 310 is, for example, melted at the surface so as to bond the other porous substrate to the barrier 310.
- the other porous substrate has a fibrous density different from the fibrous density of the initial porous substrate 305.
- the volatile compound is dissipated in the thickness of the porous substrate so as to increase the volume and evaporation surface of the volatile compound.
- the non-volatile compound is retained on the surface by the barrier.
- FIG. 4 shows an embodiment of a storage and concentration device 40 of the present invention.
- This device 40 comprises:
- a fluid injection means 325 on or in the porous substrate 305, configured to carry out a plurality of injections so as to concentrate at the same location of the porous substrate 305, successively, different quantities of the same non-volatile compound,
- a means 335 for selective destocking of a non-volatile compound is provided.
- the injection means 325 is, for example, a syringe.
- This syringe makes it possible to perform a plurality of injections of a non-volatile compound in or on the porous substrate 305.
- This non-volatile compound whether transported by a volatile compound or not, is transported on the porous substrate 305 before drying at a location of the porous substrate 305.
- the transport means 330 is, for example, a syringe configured to inject into the porous substrate 305 a solvent in which at least one nonvolatile compound dissolves.
- a container 340 such as a microchannel
- the porous substrate 305 is brought into contact with the microchannel.
- a means for reducing the internal pressure, not shown, of the microchannel makes it possible to cause the flow of the solvent compound, comprising at least one non-volatile compound, from the porous substrate 305 to the microchannel.
- the selective retrieval means 335 is, for example, a pair of scissors for cutting the porous substrate 305 according to the positioning of each place where a nonvolatile compound is concentrated.
- the injection of a solvent into each of the cut parts allows the selective destocking of at least one non-volatile compound.
- FIG. 5 shows one embodiment of a storage and concentration device 50 that is the subject of the present invention.
- This device 50 comprises:
- a means 325 for injecting a fluid onto or in the porous substrate 305 configured to produce a plurality of different fluid injections of which at least one fluid comprises at least one non-volatile compound on a porous substrate 305 so as to concentrate in different places of the porous substrate 305, various non-volatile compounds or so as to displace at least one nonvolatile compound from one place to another of the porous substrate,
- a means 335 for selective destocking of a non-volatile compound is provided.
- the injection means 325 is, for example, a syringe. This syringe makes it possible to perform a plurality of injections of at least one non-volatile compound in or on the porous substrate 305. Each non-volatile compound, whether transported by a volatile compound or not, is transported on the porous substrate 305. before drying at a location of the porous substrate 305.
- the transport means 330 is, for example, a syringe configured to inject into the porous substrate 305 a solvent in which at least one nonvolatile compound dissolves.
- a container 340 such as a microchannel
- the porous substrate 305 is brought into contact with the microchannel.
- a means for reducing the internal pressure, not shown, of the microchannel makes it possible to cause the flow of the solvent, comprising at least one non-volatile compound, from the porous substrate 305 towards the microchannel.
- the selective destocking means 335 is, for example, a pair of scissors making it possible to cut the porous substrate 305 as a function of the positioning of each place where is concentrated a non-volatile compound.
- the injection of a solvent into each of the cut parts allows the selective destocking of at least one non-volatile compound.
- FIG. 6 shows a first embodiment of a microchannel 60 viewed from above.
- This microchannel 60 comprises means 605 for receiving a fluid, this fluid can be poured into the receiving means 605 by a reservoir comprising in variants a microfluidic flow control system in the receiving means 605.
- This microchannel 60 further comprises a contact zone 610 configured to be placed in contact with a porous substrate, such as a sheet of paper for example.
- microchannels can be made by a step of shaping a material.
- this formatting step is performed by:
- thermoforming or hot molding polymer ablation, or polymer molding.
- the material used can be of any type of polymer, and for example polymers such as polystyrene (PS), polycarbonate (PC), polyvinyl chloride (PVC), cyclic Olefin copolymers (COC) Poly (Metyl Methacrylate) (PMMA), Thermoset PolyEster (TPE), PolyUrethane MethAcrylate (PUMA), or Acrylonitrile Butadiene Styrenes.
- polymers such as polystyrene (PS), polycarbonate (PC), polyvinyl chloride (PVC), cyclic Olefin copolymers (COC) Poly (Metyl Methacrylate) (PMMA), Thermoset PolyEster (TPE), PolyUrethane MethAcrylate (PUMA), or Acrylonitrile Butadiene Styrenes.
- PS polystyrene
- PC polycarbonate
- PVC polyvinyl chloride
- COC cyclic Olefin cop
- the material may also be selected from photocurable or photosensitive liquids or glues, for example Norland Optical Adhesive ("NOA").
- NOA Norland Optical Adhesive
- the material is positioned on a layer of a flat substrate.
- the molded material is positioned so that the recess, created by molding, etching or machining, forms a microfluidic channel on the flat substrate side.
- FIG. 7 shows a second embodiment of a microchannel 70 seen from above.
- the microchannel 70 has a plurality of contact areas 710.
- FIG. 8 shows a third embodiment of a microchannel 80 viewed from above.
- the microchannel 80 comprises a contact zone 810 whose width increases as the distance of a means 805 for receiving a fluid increases.
- FIG. 9 shows a first sectional view of a first particular embodiment of a microchannel 90.
- This microchannel 90 is produced, for example, according to one of the techniques described in FIG. formed between the treated material 910 and the substrate 915 acts as a conduit.
- a fluid may be deposited, for example by flow, at the opening of this cavity 905.
- the microchannel 90 is thus formed at the interface of two layers of material, at least one of the two materials being a substrate.
- the volume of the microchannel is contained in the recess formed between the substrate and the other material.
- FIG. 10 shows a second sectional view of the microchannel 90 depicted in FIG. 9.
- a portion of the material 910 treated and then placed on the substrate 915 is deformed by applying pressure on one end 920.
- the microchannel 90 formed by the material 910 so as to peel the material 910 from the substrate 915.
- the material 910 forming the end 920 of the microchannel 90 is connected to the rest of the material 910 by a hinge.
- the material 910 is shape memory and resumes an initial position in contact with the substrate 915 when the pressure is released.
- the end 920 of the microchannel 90 formed by the material 910 is fixable to the substrate 915.
- FIG. 11 shows a third sectional view of the microchannel 90 described in FIGS. 9 and 10.
- a porous substrate 925 is introduced in contact with the microchannel 90 by inserting the porous substrate 925 between the substrate 915 and the material 910 forming the microchannel 90. Once the porous substrate 925 is inserted, the peeled portion of the material 910 and the substrate 915 are forced into contact with the porous substrate 925 by a fastening clip 930 surrounding the substrate 915 and the material 910 at the location of the insertion of the porous substrate 925.
- the porous substrate 925 is composed for example of one or more cellulose, nitrocellulose or cellulose acetate paper sheets, supplemented or not by other additives; a filter paper; a textile fabric; glass fibers; and generally any porous medium in which a capillary flow of liquid occurs.
- porous substrate 925 Once the porous substrate 925 has been in contact with the microchannel 90, and according to the pressure in the microchannel 90, a fluid contained in the microchannel 90 migrates towards the porous substrate 925, or a fluid contained in the porous substrate 925 migrates towards the microchannel 90.
- the flow of liquid in the porous substrate 925 can be controlled by the shape of the substrate, by solid barriers formed in situ such as, for example, hydrophobic wax, resin or polymers, by a wetting contrast such as, for example silanization, Alkyl Ketene Dimer treatment, called "AKD", or any other flow control technique in a porous medium.
- a wetting contrast such as, for example silanization, Alkyl Ketene Dimer treatment, called "AKD”
- Soaking a paper in an AKD bath causes the chemical grafting of this molecule, which then makes the paper, naturally hydrophilic, hydrophobic.
- a plasma treatment through a metal mask makes it possible to attack the grafted chemical structure and to find locally the hydrophilic character of the paper.
- hydrophilic-hydrophobic wetting contrast a technique similar to silanization coupled with UV insolation.
- FIG. 12 shows a sectional view of a manual system for varying the internal pressure of a microchannel.
- This pressure variation system comprises a syringe 1205, a reservoir 1210 and means 1215 connecting a fluid contained in the reservoir 1210 and the microchannel.
- a user presses on the moving part of the syringe 1205 the fluid or gas contained in this syringe 1205 is injected into the reservoir 1210, increasing the pressure in this reservoir 1210.
- the increasing internal pressure of the reservoir 1210 causes evacuation part of the fluid to the microchannel.
- the pressure then increases in the microchannel until the fluid contained in the microchannel is pushed towards a porous substrate which is in contact with the microchannel.
- control of the pressure of the microchannel is achieved by pressure controllers, syringe pumps or other flow control systems.
- FIG. 13 shows a sectional view of a particular embodiment of a microchannel 1300.
- a porous substrate 1305 is inserted into a cavity 1310 formed between a treated material 1315 and a substrate 1320 through an opening in the material 1315. This opening is made by incision of the material 1315.
- FIG. 14 shows a sectional view of a second particular embodiment of a microchannel 1400.
- a porous substrate 1405 is inserted into a cavity 1410 formed inside a 1415 treated material fixed on a substrate 1420 by incision of the material 1415 and insertion of the porous substrate 1405 into the incision made.
- the microchannel 1400 is permanently fixed to the porous substrate 1405. This permanent fixing is achieved, for example, by molding the material 1415 around the porous substrate 1405. This attachment can be made, for example, by bonding the substrate. porous 1405 to material 1415.
- the purpose of bringing a microchannel into contact with a porous substrate is to be able to transfer a liquid sample from one system to another.
- the contacting can be maintained by tools to maintain a certain seal, such as fixing clips as described in Figure 1 1.
- FIG. 15 shows, viewed from above, a microchannel 1500 comprising a plurality of means 1505 for receiving a fluid, each fluid being directed as a function of the pressure applied to the fluid.
- FIG. 16 shows, viewed from above, a microchannel 1600 comprising a plurality of porous subtrates 1605 each containing at least one fluid, each fluid being sucked into the microchannel 1600 by regulating the internal pressure of the microchannel 1600.
- FIG. 17 shows, in section, a particular embodiment of an analyte 1700 retrieval kit.
- This analyte retrieval kit 1700 comprises a microchannel 1705 and a porous substrate 1710 comprising dried analytes. Dried analytes are obtained by evaporation of a solvent transporting the analytes through a porous substrate 1710.
- the porous substrate 1710 is contacted with the microchannel 1705 and a solvent 1720 is injected in the porous substrate 1710.
- the solvent 1720 is distributed in the porous substrate 1710, carrying the dried analytes during its passage.
- the pressure in the microchannel 1705 is decreased so that the analyte transported by the solvent 1720 enters the microchannel 1705. This analyte penetrates more or less into the microchannel 1705 depending on the pressure exerted.
- FIG. 18 shows, in section, a particular embodiment of a kit for storing or retrieving analytes 1800.
- a porous substrate 1805 is embedded inside a microchannel. 1810.
- the microchannel 1810 has two openings 1815, one upstream of the porous substrate 1805 and one downstream of the porous substrate 1805.
- the insertion into a first opening of a solvent comprising an analyte makes it possible to store the analyte in the porous substrate 1805 in a dry manner by evaporation of the solvent.
- a solvent is introduced through one of the openings 1815 of the microchannel 1800 so that the analyte dissolves in the solvent and leaves the microchannel 1800 through the second opening 1815.
- the Porous substrate 1805 serves as a storage matrix.
- the dry or wet chemical modification of the surfaces of the porous substrate 1805 and / or the microchannel 1810 makes it possible to control the hydrophilic or hydrophobic affinity for the samples used.
- microfluidic chip When several microchannels are mounted on a substrate, it is called "microfluidic chip".
- the microchannels of a microfluidic chip have, for example, a length of, for example, between 0.1 nanometers and several centimeters.
- the channel formed in a porous substrate may be of a size similar to that of the microchannel or very different depending on the technological means available.
- a microchannel can be manufactured, as an indication, as follows:
- the photolithography of a resin deposited on a silicon wafer makes it possible to obtain a solid and reusable mold.
- a polymer of PolyDiMethylSiloxane type, abbreviated "PDMS”, and a crosslinking agent are mixed and poured onto the mold.
- the polymer thus molded is then dissociated from the silicon mold, by slight mechanical deformation, and an inlet is pierced by a punch.
- the polymer, containing the microchannel is then glued on a glass slide by plasma treatment under oxygen.
- the glass slide may have been pre-coated with a polymer layer.
- a drying step in a silane atmosphere may precede the bonding of the polymer with the glass slide, in order to functionalize the surfaces with a chemical group present on the silane.
- the PDMS mold can be used as a stamp to transfer the pattern to a photo-crosslinkable glue after ultraviolet irradiation.
- a microfluidic paper chip can be made, as an indication, as follows:
- the paper microfluidic system can result from a simple cutting of the substrate.
- Paper channels can be defined by wetting contrast.
- a prior chemical treatment (AKD type or silanization) makes it possible to control the state of the entire substrate.
- Ultraviolet insolation or plasma treatment located through a mask made of metal, quartz, resin, chromium or plastic for example or by focusing makes it possible to change the state locally.
- the channels on paper can be defined by in situ solidification of a compound making it possible to block the pores of the porous substrate: such as a wax, a polymer or a resin.
- a compound making it possible to block the pores of the porous substrate: such as a wax, a polymer or a resin.
- the location of these barriers can result from specific insolation (photolithography) or from a specific deposit (solid ink printer for example).
- the formation of microstructures dug, for example by a laser, can also constitute a form of barrier.
- the paper support may also have been incised so as not to form a porous wetting medium but a single channel.
- the contacting between the microchannel and the porous substrate is carried out, for example, as follows:
- the microfluidic paper system and the microchannel can be brought into contact by specifically detaching the two constituent layers of the material and the substrate with a scalpel.
- the layers are of polymer, or polymer-glass, or photocrosslinkable-glass glue, or photocrosslinkable glue.
- the porous substrate is then slid between the two constituent layers.
- a fixing clip is positioned so as to exert a pressure on both sides of the two constituent layers to restore the seal.
- the porous substrate may also be slid into a slot, horizontal, vertical or oblique, made within the constituent polymer of the microchannel.
- the contacting between the porous substrate and the microchannel can be performed during manufacture of the microchannel by sliding the porous substrate between the two constituent layers before or during the bonding step, or by molding a portion of the microchannel around the porous substrate.
- FIG. 19 shows a porous substrate 1900 comprising an injection-recovery zone 1905 on which is deposited a solvent comprising dissolved analytes. In FIG. 19, the case in particular is observed where the flow velocity of the analytes is lower than the flow rate of the solvent.
- the solvent forms a solvent front 1910 and the position of the analytes is bounded between this solvent front 1910 and the injection-recovery zone 1905.
- a porous substrate 2000 as described is observed. in Figure 19 wherein the flow rate of the analytes is zero. In this configuration, the analytes remain at the 2005 injection-recovery zone.
- Evaporation of the solvent is naturally present because of the large air / liquid free interfaces, the microfluidic channels being open.
- an enrichment process which consists of making several successive solvent deposits in which analytes are dissolved, separated by waiting times to allow evaporation, provides a concentration effect by increasing the amount of analyte deposited . This phenomenon is interesting from the moment when the injection-recovery zone is controlled, for example with a view to a selective destocking or a reaction with a reagent whose position is specific.
- FIG. 21 shows a porous substrate 2100 comprising a zone 2105 for injection-recovery of solvent comprising an analyte 21 of interest.
- FIG. 22 the porous substrate 2100 depicted in FIG. 21 is seen in which solvent has been added to the injection-recovery zone so as to displace the analyte 21 on the porous medium.
- FIG. 23 shows a curve representative of the local concentration of analyte in a function of the number of successive solvent deposits on the injection-recovery zone of a porous substrate. In particular, it is observed that the analyte concentration increases until a saturation threshold is reached which depends on the total quantity of analyte deposited.
- FIG. 24 shows a curve of the local concentration of analyte of interest as a function of the volume of solvent deposits comprising an analyte of interest on the injection-recovery zone and the volume of solvent deposited in the zone. injection-recovery.
- the method consists in limiting the spreading of the drop on the surface to restrict the injection-recovery zone while allowing the capillary pump to extract the solvent. Without barriers, the drop spreads over a large area. With barriers throughout the thickness of the porous substrate, the drop is well retained spatially but the evaporation time is long because the drop remains in the form of spherical cap. With barriers on the surface or in a partial thickness of the porous substrate, there is no spreading of droplets, so the injection-recovery zone is restricted, the capillary pump makes it possible to extract the solvent. The drop is transformed into a thin film which has a much shorter evaporation time. FIG.
- FIG. 25 shows a sectional view of a particular embodiment of a deposit of a 2505 drop of a solvent comprising an analyte on a porous substrate 2510 having no barrier.
- FIG. 26 shows a view from above of the depot illustrated in FIG. 25. In this FIG. 26, it can be seen in particular that the injection-recovery zone of the analyte 2605 and the injection-recovery zone of the solvent 2610 are of similar size. In this configuration, evaporation of the solvent is rapid but the injection-recovery zone of the analyte is wide.
- FIG. 27 shows a sectional view of a particular embodiment of a deposit of a 2705 drop of a solvent comprising an analyte on a porous substrate 2710 comprising barriers 2715 on the surface.
- FIG. 28 shows a top view of the depot illustrated in FIG. 27. In this FIG. 28, it can be seen in particular that the injection-recovery zone of the analyte 2805 bounded by the barrier 2815 forms a closed surface. , is much smaller in size than the 2810 solvent injection-recovery zone, which is not limited by the barrier. In this configuration, evaporation of the solvent is rapid and the area of injection-recovery of the analyte is small.
- FIG. 28 shows a sectional view of a particular embodiment of a deposit of a 2705 drop of a solvent comprising an analyte on a porous substrate 2710 comprising barriers 2715 on the surface.
- FIG. 28 shows a top view of the depot illustrated in FIG. 27. In this FIG. 28, it can
- FIG. 29 shows a sectional view of a particular embodiment of a deposition of a drop 2905 of a solvent comprising an analyte on a porous substrate 2910 comprising barriers 2915 in the thickness of the substrate porous.
- FIG. 30 shows a top view of the depot illustrated in FIG. 29. In this FIG. 30, it can be seen in particular that the injection-recovery zone of the analyte 3005 is of a size similar to the size of the zone. In this configuration, the evaporation of the solvent is slow but the injection-recovery zone of the analyte is small.
- analyte By repeating the deposition process on a porous substrate with surface barriers a large number of times, a large amount of analyte can be deposited on the injection-recovery zone in a relatively short time because of the short time required. 'evaporation. The deposited analyte concentration thus increases linearly with the deposited sample volume.
- FIG. 31 shows a sectional view of a particular embodiment of a barrier 3105 on a porous substrate 31 in which the barrier is a hydrophobic adhesive tape.
- FIG. 32 shows a sectional view of a second particular embodiment of a barrier 3205 on a porous substrate 3210 in which the barrier is a wax deposited in a small amount on the porous substrate 3210.
- Fig. 33 is a sectional view of the second particular embodiment of the barrier 3305 as described in Fig. 32 wherein the wax has been heated to penetrate a porous substrate 3310 to form a barrier therein.
- FIG. 34 shows a sectional view of a third particular embodiment of a barrier 3405 whose thickness varies in the porous substrate 3410. In this configuration, the concentration of analyte is carried out both in the direction of the flow and transversely in the porous substrate.
- the device comprises an injection-recovery zone, delimited at least partially by a barrier, and one or more ends.
- the sample used contains at least two analytes: one well retained by the porous substrate, the other well transported by the flow.
- the sample is deposited in one or more volumes on the injection-recovery zone, then one or more solvent volumes are added.
- the well-retained compound is concentrated before the surface barrier, the transported compound is deposited on the end.
- the separation and concentration steps are performed simultaneously. It is possible, in addition, to use a plurality of analytes moving with the solvent with varying speeds so as to separate and spatially concentrate several analytes with a single device.
- FIG. 35 shows a particular embodiment of a device 3500 for separating and concentrating a plurality of analytes, which comprises a porous substrate 3505 in which barriers 3510 throughout the thickness of the porous substrate allow to guide the flow.
- This device 3500 further comprises an injection-recovery zone 3515 of a plurality of analytes dissolved in one or more solvents delimited by a barrier 3520 at the surface or at a partial thickness to effect separation and concentration.
- a method of concentrating a flow-transported sample in a porous medium can be carried out as follows:
- the method of concentrating a sample well transported by the flow comprises a step of depositing the sample followed by additions of solvent - water in the case of hydrophilic compounds.
- the porous substrate is treated by a wetting contrast, or solid barriers, or a blank, or any other manufacturing method, so as to have a closed channel type geometry at the end. It may also have an injection-recovery zone connected to the channel.
- a sample volume is deposited on the injection-recovery zone.
- the liquid flows in all accessible geometry, thanks to the capillary pump. Large free interfaces between liquid and air facilitate evaporation.
- the analyte dissolved in the solvent is then deposited in dried form over the entire accessible surface of the porous medium.
- the analyte is dried and is again dissolved and transported a certain distance, before being deposited in dried form, again.
- the volume of solvent required depends on the porosity of the porous and the geometry of the channel.
- a method of concentrating a sample retained in a porous medium can be performed as follows:
- the method of concentrating a sample well retained by the porous medium comprises several successive sample deposits.
- the porous substrate comprises an injection-recovery zone delimited by a barrier. This barrier can be made by a piece of tape or by depositing a small amount of wax.
- the sample is deposited on the injection-recovery zone.
- the capillary pump extracts the solvent, and the evaporation is fast thanks to this extraction.
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Abstract
Description
Claims
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BR112016010225-8A BR112016010225A2 (pt) | 2013-11-08 | 2014-11-07 | método e suporte para armazenar e concentrar um composto não volátil. |
EP14809478.2A EP3065870A2 (fr) | 2013-11-08 | 2014-11-07 | Procédé de stockage et de concentration d'un composé non volatil |
US15/034,524 US20160327460A1 (en) | 2013-11-08 | 2014-11-07 | Method and support for storing and concentrating a non-volatile compound |
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FR1360987 | 2013-11-08 | ||
FR1360987A FR3012982B1 (fr) | 2013-11-08 | 2013-11-08 | Procede de stockage et de concentration d'un compose volatil |
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WO2015067912A2 true WO2015067912A2 (fr) | 2015-05-14 |
WO2015067912A3 WO2015067912A3 (fr) | 2015-07-02 |
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PCT/FR2014/052866 WO2015067912A2 (fr) | 2013-11-08 | 2014-11-07 | Procédé de stockage et de concentration d'un composé non volatil |
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US (1) | US20160327460A1 (fr) |
EP (1) | EP3065870A2 (fr) |
BR (1) | BR112016010225A2 (fr) |
FR (1) | FR3012982B1 (fr) |
WO (1) | WO2015067912A2 (fr) |
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FR3117897A1 (fr) * | 2020-12-21 | 2022-06-24 | Commissariat à l'Energie Atomique et aux Energies Alternatives | Procédé de récupération d'un échantillon capturé dans une membrane hydrophile et dispositif destiné à mettre en œuvre le procédé |
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2013
- 2013-11-08 FR FR1360987A patent/FR3012982B1/fr active Active
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2014
- 2014-11-07 EP EP14809478.2A patent/EP3065870A2/fr not_active Withdrawn
- 2014-11-07 BR BR112016010225-8A patent/BR112016010225A2/pt not_active Application Discontinuation
- 2014-11-07 US US15/034,524 patent/US20160327460A1/en not_active Abandoned
- 2014-11-07 WO PCT/FR2014/052866 patent/WO2015067912A2/fr active Application Filing
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Also Published As
Publication number | Publication date |
---|---|
EP3065870A2 (fr) | 2016-09-14 |
US20160327460A1 (en) | 2016-11-10 |
FR3012982A1 (fr) | 2015-05-15 |
WO2015067912A3 (fr) | 2015-07-02 |
FR3012982B1 (fr) | 2015-12-25 |
BR112016010225A2 (pt) | 2018-05-02 |
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