Method and device for extracting substances from adsorption layer of chromatography plate

Abstract

The invention relates to a method and a device for extracting substances from an adsorption layer of a chromatography plate. According to the invention, the method comprises forming a microcontainer, the side wall of which is a sleeve wall vertically passing through the adsorption layer and pressed against the carrier plate of the chromatographic plate, and the bottom of which is the carrier plate of the chromatographic plate. One or more dozens to hundreds of microliters of extract liquor are added to the microcontainer formed in this way. And after the extraction liquid is extracted from the sleeve, qualitative and / or quantitative analysis is carried out by using an instrument technology. The method is carried out by adopting a mechanism for vertically pressing the sleeve to the chromatoplate. The sleeve has a lower edge with a special profile, which helps to achieve a tight connection between the lower portion of the sleeve and the chromatography plate carrier plate. The device uses a replaceable sleeve made of a chemically resistant material, preferably plastic, such as polypropylene.

Description

[0001] The subject of this invention is a method and apparatus for extracting substances from specific regions / spots in an adsorption layer on a chromatographic plate, the regions / spots being obtained after chromatographic development, and containing one or more components derived from a separated mixture. The obtained extract can be used for qualitative and / or quantitative analysis using instrumental techniques.

[0002] In laboratory practice, the simplest method for extracting a substance from the adsorption layer of a chromatographic plate is to scrape off a specific area containing the substance or mixture of substances, transfer it to a filter paper layer placed on a funnel, and elute the substance with a suitable solvent. Depending on its intended use, such as for quantitative or qualitative analysis or for preparative purposes, the resulting extract may be further processed. This method is described in the monograph on thin-layer chromatography, *Thin-Layer Chromatography, Fourth Revision and Expanded Edition*. It is described in Chapters 10 and 12 of (Thin-Layer Chromatography, Fourth Edition, Revised and Expanded, B. Fried, J. Sherma, 1999, Marcel Dekker, Inc.).

[0003] In B. Falk and K. Krummen's publication J. Chromatogr. A, 103 (1975), 279–288, a method and apparatus for eluting substances from spots on a chromatographic plate are described. The essence of this method and apparatus is to form an annular groove in the adsorption layer surrounding the spot, press a Teflon head with a seal onto this groove, and use a syringe pump to supply eluent to one opening of the head and collect the eluent through another opening. A suitably selected solvent flows through the adsorption layer region defined by the aforementioned seal, eluting the substance therefrom. Using this apparatus, substances can be eluted from six spots in the adsorption layer of a chromatographic plate simultaneously. The disadvantages of this apparatus are that the groove for the seal on the head needs to be manually made in the adsorption layer, and the apparatus needs to be flushed before the next elution cycle. Furthermore, the use of a syringe pump adds to the cost of the apparatus.

[0004] A known apparatus for eluting substances from an adsorption layer on a chromatographic plate is described in H. Luftmann's publication Anal. Bioanal. Chem. (2004) 378: 964–968, DOI 10.1007 / s00216-003-2293-3 and patent specification DE10036293A1. The apparatus comprises a steel piston with a sharp edge along its entire circumference on its lower surface and two capillary channels, one for supplying the extract and the other for discharging it. The extract is supplied via a pressure pump. To elute the substance from the adsorption layer, the piston is pressed against the layer, and the eluent is forced through the supply channel, the adsorption layer, and the discharge channel by the pressure pump. From the latter channel, the liquid containing the eluted substance is directed to a vial or directly into the analytical instrument. The disadvantages of the proposed method are that the apparatus channels are frequently clogged by adsorbent particles, requiring thorough cleaning before proceeding to the next elution step from the next position on the chromatographic plate adsorbent layer, and the use of a pressure pump, which is typically expensive equipment. This method also consumes a large amount of eluent, and the obtained sample is diluted, sometimes requiring concentration for subsequent steps.

[0005] According to US Patent 5208458, dated May 4, 1993, an apparatus is known for connecting various types of planar electrophoresis maps, including gel electrophoresis maps, to a mass spectrometer. This connection apparatus includes: a pump that delivers solvent to the surface of the electrophoresis map at a predetermined rate; a system for breaking / crushing the gel and releasing the molecules contained therein so that they can be absorbed by the solvent; and a capillary for transferring the solvent containing the eluted material to the mass spectrometer. The limitation of this apparatus is that it is intended for use with electrophoresis maps from which material elutes into the mass spectrometer. Furthermore, the elution solvent must be supplied by a separate pump connected to the sample injector of the mass spectrometer, making the apparatus complex, relatively expensive, and prone to clogging of the connection channels.

[0006] Another solution is described in US patent application US 2011 / 0269166 A1, dated November 3, 2011. The sample, located in an adsorption layer (acting as a filter layer), is placed on a flexible hydrophobic substrate. The sample may contain biological or chemical material as well as an absorbent / filter material / layer. The filter layer contains absorbent material embedded with the sample material. A sampler is used to collect the sample using a sampler that conforms its surface to the absorbent material. The sampler has a sharp edge pressed against the hydrophobic substrate, ensuring a tight bond between the sampler and the flexible hydrophobic substrate and ensuring that the elution solvent flows under pressure through the absorbent / filter material. The sampler has two supply and discharge channels. Substances in the sample are eluted from the adsorption layer using an additional pump, which forces the elution solvent into the supply channel, then through the absorbent / filter layer, and out of the sampler through the discharge channel. The effluent from the discharge channel can be collected in a vial or introduced directly into the instrument analysis device. The sampler has some drawbacks, such as the discharge channel being prone to clogging, the sampler needing to be washed / cleaned after each sample elution process, and the use of an additional pump increasing the cost of using the sampler.

[0007] US Patent Application No. 2012 / 0125127 A1, published on May 24, 2012, describes an apparatus for preparing samples for qualitative and quantitative instrumental analysis. This apparatus allows a porous material containing adsorbed sample components to be enclosed in a chamber formed between two compressed blocks. The connection between the compressed blocks is tight, allowing elution solvent to be supplied to the material enclosed in the chamber under pressure, eluting the substance into vials or instrumental analysis devices. The apparatus must be connected to a pressure pump, which significantly increases its cost. Furthermore, during use, the channels of the apparatus may become clogged with particles of the porous material, and the chamber and channels of the apparatus require thorough cleaning before each subsequent use with a new sample.

[0008] The purpose of this invention is to provide an apparatus for extracting substances from the adsorption layer of a chromatographic plate, wherein the channels of the apparatus are not blocked by adsorbent particles. Furthermore, the invention aims to provide an easy-to-use apparatus that does not require washing before proceeding to the next elution step from the next location on the adsorption layer of the chromatographic plate, and an apparatus that does not require the use of a pressure pump (typically expensive equipment). Additionally, the invention aims to provide an apparatus that requires a small amount of eluent, and the obtained sample is not over-diluted, nor does it require concentration in subsequent steps. The proposed invention solves the problem of extracting substances from specific locations in the adsorption layer of a chromatographic plate. The essence of the method for extracting substances from specific locations in the adsorption layer of a chromatographic plate (typically the region / spot containing the substance after chromatogram development) lies in using a small-capacity sleeve with a specially contoured, sharp tip, pressed vertically through the adsorption layer onto a carrier plate (the carrier plate with the adsorption layer) with minimal force. The sleeve is pressed against the chromatographic plate, thereby forming a miniature container whose bottom is the carrier plate of the adsorption layer, and whose walls are the walls of the sleeve. According to the proposed invention, a predetermined small volume of extract is introduced into a formed micro-container through the upper opening of a sleeve using a syringe or pipette. The extract is then kept in contact with the adsorption layer for a specified short time. The extract containing the extracted substance is then withdrawn from the micro-container using a syringe or pipette. The obtained extract is placed in a vial, and the solid particles in the adsorption layer are either allowed to settle or removed by filtration or centrifugation before being placed in the vial, and then sent for instrumental analysis.

[0009] A method for extracting substances from the adsorption layer of a chromatographic plate / from a specific location in the adsorption layer of a chromatographic plate, characterized by comprising the following steps: A micro-container is formed between a carrier plate of a chromatographic plate and a sleeve pressed onto it, wherein the sleeve is pressed at a right angle onto the carrier plate with an adsorption layer, such that the bottom of the micro-container is the carrier plate with the adsorption layer, and the wall of the micro-container is the wall of the sleeve, wherein the height of the micro-container is preferably up to 50 mm. The carrier plate with the adsorption layer is referred to as a chromatographic plate. The sleeve is pressed onto the chromatographic plate to form a micro-container, the height of which is preferably up to 50 mm, more preferably between 5 mm and 50 mm, and even more preferably between 10 mm and 50 mm; A predetermined small volume of extract, greater than 20 μL, preferably between 20 μL and 500 μL, is introduced into the resulting micro-container through the upper opening of the sleeve. Using such a volume of extract (preferably greater than 20 μL), the liquid can be freely collected from above the adsorption layer using a pipette. Simultaneously, some liquid is not absorbed into the adsorption layer. The extract can be advantageously introduced using a syringe or pipette, in volumes ranging from tens to hundreds of microliters; • The extract is kept in contact with the adsorption layer in a micro-container for a specified short time, ranging from 1 minute to 10 minutes. Advantageously, the contact time between the extract and the adsorption layer can be from 10 seconds to 10 minutes; for screening analysis, this time is preferably from about 10 seconds to 1 minute. The incubation time can also be from 1 minute to 10 minutes. • Extract the extract (i.e., the extract containing the extracted substance) from the microcapsule. Advantageously, use a syringe or pipette to remove the liquid from the microcapsule.

[0010] Preferably, the method is characterized in that, in a subsequent stage, the extract is sent for instrumental analysis, or the solid particles of the adsorbent layer are separated from the extract before being sent for instrumental analysis.

[0011] Preferably, the method according to the invention is characterized by including the step of removing solid particles from the adsorbed layer, wherein the solid particles are removed from the extract by filtration or centrifugation before the extract is placed in the vial, or by allowing the solid particles to undergo a sedimentation process after placement in the vial.

[0012] Preferably, the method according to the invention is characterized in that the micro-container has a circular internal opening with an inner diameter preferably of 2 mm to 10 mm, or has an elliptical internal opening with a minor axis dimension preferably of 2 mm to 6 mm and a major axis dimension of 3 mm to 10 mm.

[0013] Preferably, the method according to the invention is characterized in that a portion of the introduced extract is extracted from the microcontainer and immediately reintroduced into the microcontainer, and the operation of extracting and introducing the portion of extract is repeated 2 to 10 times.

[0014] Preferably, the method according to the invention is characterized in that the extraction process is repeated several times using a new portion of the extract, the obtained extracts are combined, and the number of extraction repetitions is determined experimentally.

[0015] A specific amount of extract is introduced into a microcontainer, and preferably using a syringe or pipette, a portion of it is withdrawn from the microcontainer and immediately reintroduced, repeated multiple times within short time intervals. The number of these withdrawal and introduction operations, and the duration of these operations, are determined experimentally. Immediate withdrawal and introduction of the extract is equivalent to a mixing process. It is advantageous to use the same liquid during both withdrawal and introduction, which mixes it and accelerates extraction and the establishment of thermodynamic equilibrium.

[0016] As an alternative to performing the above-described repeated removal and introduction of a portion of the extract, the extract in the micro-container can be stirred.

[0017] An advantageous feature of the method according to the invention is that the extract in the micro-container is stirred, advantageously using a micro-stirrer or by shaking or ultrasonic treatment.

[0018] An advantageous feature of the method for extracting substances from a chromatographic plate according to claim 1 is that the extraction time for a single extraction process is up to 10 minutes, and is advantageously determined experimentally.

[0019] An advantageous feature of the method according to the invention is that, during the extraction process, the upper opening of the sleeve is advantageously sealed with a stopper / cork with an opening.

[0020] The phrase "during the extraction process" is equivalent to "during the extraction process" or "during extraction." This implies that the substance is transferred from the adsorption layer into the extraction solvent.

[0021] An apparatus for extracting substances from a chromatographic plate, comprising a body and a stage for placing the chromatographic plate, characterized in that it includes: • Components connected to the body for securing at least one sleeve in the form of a bracket and / or fastening beam; • A pressure-applying mechanism connected to the main body of the apparatus ensures that at least one sleeve is pressed vertically onto the chromatographic plate and is capable of removing at least one sleeve from the chromatographic plate; • Elastic elements ensure proper fit between the sleeve edge and the chromatographic plate.

[0022] The main body can be a single component or multiple components.

[0023] The stage uniformly supports the chromatographic plate at the extraction site, and the stage can be integrally formed with the main body (as a single component), or the stage can be an additional component directly or indirectly connected to the main body.

[0024] When directly connected to the body, the pressure mechanism, along with the sleeve, moves toward the chromatographic plate placed on the worktable. When the pressure mechanism is located below the worktable, the worktable, along with the chromatographic plate, is moved and pressed against the sleeve. The worktable can be directly connected to the body, integrally formed with it (it can be part of the body), or the worktable can be indirectly connected to the body, for example, through the pressure mechanism.

[0025] The fastening elements allow for interchangeable vertical fixation of at least one sleeve. The sleeve can be mounted in a bracket connected to, for example, a main body / frame beam, or directly in a pressure beam. The method of connecting the sleeve to the bracket can be arbitrary, as long as a stable connection and uniform fixation of the sleeve are ensured.

[0026] The bracket can be equipped with a snap-fit ​​mechanism for the fixed sleeve, and the bracket can be connected to the pressure application mechanism via an elastic element.

[0027] The sleeve support also functions through a mounting beam, which is preferably equipped with an O-ring mounted in the pressure beam for securing at least one sleeve. The shape of the mounting beam and the arrangement of the sleeve-fixing elements can be arbitrary.

[0028] The pressure mechanism ensures that at least one sleeve is pressed vertically onto the chromatographic plate and allows the sleeve to be moved a certain distance from the chromatographic plate for easy replacement, that is, the sleeve is lifted at least a few millimeters above the chromatographic plate.

[0029] The pressure application mechanism can be directly or indirectly connected to the body of the apparatus, wherein the pressure application mechanism can be located below the stage and lift the stage with the chromatographic plate to at least one sleeve, or attached to an element that fixes at least one sleeve, and then the mechanism and the sleeve are moved together to the chromatographic plate.

[0030] The pressure applying mechanism can adjust the pressure via a turnbuckle, or it can include an additional element that adjusts the pressure based on strain gauge sensor readings by controlling the operation of the pressure applying mechanism.

[0031] The elastic element ensures that the edge of the sleeve conforms to the chromatographic plate. For example, the elastic element is a pair of elastic bodies connected to the pressure application mechanism. Alternatively, the elastic element is an O-ring, which additionally performs the function of fixing the sleeve.

[0032] Advantageously, the device according to the invention is characterized in that it has at least one replaceable sleeve made of a chemically inert material (preferably plastic) that is sharpened at its end along the entire circumference of its lower edge, and the sharp end is formed by the connection of two surfaces of the sleeve (i.e., the outer surface of the sleeve and the inner chamfered surface of the sleeve), or the edge is formed by two surfaces: one is the surface of the inner wall of the sleeve and the other is the outer chamfered surface of the bottom of the sleeve, or the edge is formed by the outer chamfered and inner chamfered surfaces.

[0033] Advantageously, the apparatus for extracting substances from a chromatographic plate is characterized in that the angle between the surfaces forming sharp edges is an acute angle, ranging from 15° to 70°.

[0034] The sleeve with sharp edges according to the present invention can apply pressure evenly on the circumference and achieve a tight connection between the sleeve and the chromatographic plate carrier.

[0035] Advantageously, the device is characterized in that the edge formed at the junction of the outer surface of the sleeve and the inner chamfered surface of the sleeve has a passivation treatment over the entire circumference, and advantageously, the width of the passivation surface is 0.1 to 0.5 mm.

[0036] Advantageously, the device is characterized in that the sleeve has at least one hole on its wall near the end opposite to the end with the sharp edge, the hole having a diameter that is advantageously in the range of 0.3 to 1.5 mm.

[0037] This opening is optional and is used to equalize the pressure inside and outside the sleeve when the pipette is inserted into the sleeve, fitted to it, and the solution is withdrawn. It can be used as an option when using a manual extractor, as the user may use different pipettes. However, in the case of an automated extractor, a standard pipette will be used, so the presence or absence of this opening is less important.

[0038] Preferably, the device is characterized in that the sleeve has at least one longitudinal groove on its inner wall, starting from the end opposite to the end of the sleeve with a sharp edge, with a length of up to 10 mm, a width of 0.3-1.0 mm, and a depth of 0.3-0.6 mm.

[0039] Advantageously, the device is characterized in that the inner cross-section of the sleeve is circular, with a diameter preferably between 2 and 10 mm, or the inner cross-section of the sleeve is elliptical, with the minor axis preferably in the range of 2 to 6 mm and the major axis preferably in the range of 3 to 10 mm, or the inner cross-section of the sleeve has a shape that is close to circular or elliptical.

[0040] Advantageously, the device is characterized in that the length of the sleeve can be up to 50 mm, preferably between 5 mm and 50 mm, and more advantageously between 10 mm and 50 mm.

[0041] Advantageously, the device is characterized in that the sleeve has one or two protrusions / flanges on its outer surface around its entire circumference, or has one or two grooves on its entire circumference.

[0042] Advantageously, the device is characterized in that the sleeve is advantageously provided with a plug at the hole at the end opposite to the end with the sharp edge, and the plug is equipped with a hole with a maximum diameter of 2 mm.

[0043] Advantageously, the device is characterized in that the handle is equipped with a snap-fit ​​mechanism for a retaining sleeve, wherein the handle is connected to the pressure-applying mechanism via an elastic element.

[0044] Advantageously, the device is characterized in that the sleeve support is an element in the form of a fastening beam, and is advantageously equipped with an O-ring for fixing the sleeve.

[0045] Advantageously, the device is characterized in that the element of the pressure-applying mechanism is a turnbuckle.

[0046] Advantageously, the device is characterized in that the pressure-applying mechanism includes a turnbuckle that allows force adjustment, a lever, an upper pressure plate, and a lower pressure plate connected to an elastic element, which is advantageously a pair of elastic bodies that connect the pressure-applying mechanism to the sleeve support.

[0047] The lever can be operated by the operator. In this type of scheme, the support, along with the sleeve, moves toward the chromatographic plate due to the use of this pressure mechanism.

[0048] Preferably, the device is characterized in that the pressure applying mechanism is connected to the mounting beam via a strain gauge sensor and ensures that the mounting beam with at least one sleeve is displaced and that at least one sleeve is pressed against the chromatographic plate, or the pressure applying mechanism is also connected to the worktable via a strain gauge sensor and ensures that the worktable with the chromatographic plate is displaced toward at least one sleeve.

[0049] Preferably, the device is characterized in that it includes an additional element that uses a system that controls the operation of the pressure application mechanism based on strain gauge sensor readings to regulate the pressure.

[0050] The support / beam is connected to the pressure application mechanism via a strain gauge sensor, allowing at least one sleeve to be pressed onto the chromatography plate with controlled force.

[0051] Advantageously, the device is characterized in that the worktable is directly connected to the body, or the worktable is connected to the pressure applying mechanism via a strain gauge sensor, wherein the pressure applying mechanism is connected to the body.

[0052] Advantageously, the device is characterized in that the fastening beam is connected to a rotating mechanism that enables the beam and the sleeve thereon to rotate 180°.

[0053] The crossbeams for fastening the sleeve and the main body / frame can be installed in a rotatable manner, which is particularly helpful for changing the sleeve.

[0054] Advantageously, the device is characterized in that the elastic element is a pair of elastic bodies placed in the pressure-applying mechanism, and / or the elastic element is an O-ring that simultaneously fixes the sleeve.

[0055] Advantageously, the device is characterized by having a rigid frame platform with rigid support brackets on both sides, above which a rigid frame beam is horizontally attached, and includes a sleeve bracket in which sleeves are interchangeably attached vertically. Two elastic washers, each a few millimeters thick, are attached to the upper left and right sides of the sleeve bracket, above which a lower pressure plate is connected. One end of a turnbuckle is attached from its upper side to the lower pressure plate, and the other end is attached to an upper pressure plate, which is connected to the arm of a pressure lever attached to the frame beam. The lower pressure plate is slidably connected to the frame beam in a vertical direction, wherein the distance between the lower and upper pressure plates is adjusted by the shortening / extending range of the turnbuckle, and the movement of the lower pressure plate is restricted to the distance between a lower limit bar and an upper limit bar connected to the frame beam. Two springs are placed between the lower limit bar and the lower pressure plate to support the upward movement of the lower pressure plate, along with the sleeve bracket, turnbuckle, and upper pressure plate, when the pressure lever arm is raised. The length of the sleeve is preferably 10 to 50 mm.

[0056] The sleeve has one or two protrusions / flanges on its outer surface around the entire circumference.

[0057] Preferably, the sleeve has a plug at the opening at the end opposite to the end with the sharp edge, and the plug has a hole with a diameter preferably up to 2 mm.

[0058] In an apparatus for extracting substances from a chromatographic plate, the plate is placed on a rigid frame platform with the adsorption layer facing upwards. Two rigid supports are mounted on opposite sides of this platform. A rigid frame beam is horizontally attached above the platform. The apparatus presses a sleeve vertically against the plate and lifts it above the plate. The sleeve is made of a chemically inert material (preferably plastic, such as polypropylene) and is interchangeably placed in a sleeve holder connected to a mechanism for pressing the sleeve vertically against the plate from the adsorption layer side and lifting it at least a few millimeters above the adsorption layer. The sleeve holder is connected to a flexible pad a few millimeters thick, to which a lower pressure plate is attached. The lower pressure plate is slidably vertically connected to the frame beam and to an upper pressure plate via a turnbuckle. The upper pressure plate is connected to an arm of a pressure lever attached to the frame beam. The arm of the pressure lever applies pressure sequentially to the upper pressure plate, the turnbuckle, the lower pressure plate, the flexible pressure pad, the sleeve holder, and the sleeve, with the sleeve pressing directly against the plate with its sharp edge. The frame beam is equipped with a lower limit bar and an upper limit bar, which restrict the movement of the lower pressure plate. Two springs are located on the left and right sides between the lower pressure plate and the lower limit bar; these springs support the upward movement of the lower pressure plate when the pressure lever arm is lifted. Then, along with the lower pressure plate, the sleeve support, sleeve, turnbuckle, and upper pressure plate are also lifted. The distance between the lower and upper pressure plates, and thus the force exerted by the sleeve against the chromatographic plate, is adjusted using the turnbuckle. The lower end of the sleeve, which is in direct contact with the chromatographic plate, has a sharp edge along its entire circumference, formed by the outer surface of the sleeve and the chamfered surface inside the sleeve. The angle between these surfaces is acute, ranging from 15° to 70°. The solution according to the invention has many advantages. The solution according to the invention is characterized by its very simple and reliable operation, and the elimination of the need for an expensive liquid pump. Each extraction process from a specific location on the chromatographic plate uses at least one sleeve. The sleeve is easy to install and remove from the device. Extraction from another location on the chromatographic plate uses a new / clean sleeve. The sleeve can be used once or repeatedly after cleaning / washing. The device requires no cleaning / washing before the next extraction cycle. Extraction uses extremely small amounts of extractant, ranging from tens to hundreds of microliters, making it very economical and environmentally friendly. The use of an elastic pressure pad in the device ensures uniform pressure on the lower edge of the sleeve and adheres it to the chromatographic plate, which improves the tightness of the connection between the sleeve and the adsorption layer carrier. The proposed shape of the sleeve's sharp edge facilitates easier sealing with the chromatographic plate because, when pressed against the plate, the diameter of the sharp edge increases slightly at the contact point, causing a minimal movement on the carrier. This results in a slight movement of the adsorption layer, thereby removing adsorbent particles from the contact point between the sharp edge and the adsorption layer carrier. This allows for a tight seal to be achieved using relatively small forces when pressing the sleeve against the chromatographic plate.Furthermore, the extraction process in the device is carried out at atmospheric pressure, which helps to use a smaller force to press the sleeve against the adsorption layer to seal the contact points between the sharp edges of the sleeve and the adsorption layer carrier.

[0059] The method according to the invention is also easy to use, requiring no rinsing device before proceeding to the next elution step from the next position on the adsorption layer of the chromatographic plate, and eliminating the need for a pressure pump. Furthermore, this method provides an apparatus that requires a small amount of eluent, and the obtained sample is not over-diluted, eliminating the need for concentration in subsequent steps.

[0060] The subject matter of the invention is illustrated in the accompanying drawings in a non-limiting embodiment, wherein the figures show:

[0061] Figure 1a. A longitudinal vertical cross-section of a sleeve with a sharp lower edge on the outer wall and a circular or elliptical internal opening.

[0062] Figure 1b. Bottom cross-sectional view of a sleeve with a sharp lower edge on the outer wall and a circular internal opening.

[0063] Figure 1c. Bottom cross-sectional view of a sleeve with a sharp lower edge on the outer wall and an elliptical internal opening.

[0064] Figure 2a. A longitudinal vertical cross-section of a sleeve with a sharp lower edge on the inner wall and a circular or elliptical internal opening.

[0065] Figure 2b. Bottom cross-sectional view of a sleeve with a sharp lower edge on the inner wall and a circular internal opening.

[0066] Figure 2c. Bottom cross-sectional view of a sleeve with a sharp lower edge on the inner wall and an elliptical internal opening.

[0067] Figure 3a. A longitudinal vertical section of a sleeve with a sharp lower edge between the outer and inner walls and an internal opening that is circular or elliptical.

[0068] Figure 3b. Bottom cross-sectional view of a sleeve with a sharp lower edge and a circular internal opening between the outer and inner walls.

[0069] Figure 3c. Bottom cross-sectional view of a sleeve with a sharp lower edge between the outer and inner walls and an elliptical internal opening.

[0070] Figure 4. Schematic longitudinal vertical cross-section of a sleeve variant with a blunted sharp lower edge and an upper plug.

[0071] Figure 5. Figure 4 A magnified schematic diagram of a partial longitudinal vertical section of the outer wall of the middle sleeve, showing a blunted, sharp lower edge.

[0072] Figure 6a. Example of a sleeve cross-section marked with internal geometry and the upper edge of the beveled side.

[0073] Figure 6b. Example of a sleeve cross-section with internal geometry marked.

[0074] Figures 7a-f. The nature of the extraction method from the adsorption layer on a chromatographic plate.

[0075] Figure 8a. Front view of the apparatus in one implementation variant, with the sleeve positioned above the chromatographic plate.

[0076] Figure 8b. Front view of the apparatus sleeve pressing against the chromatographic plate.

[0077] Figure 9. Front view of apparatus variant 2 during the extraction process. Components such as the pressure beam, sleeves, and O-rings are shown in cross-sectional form on the plane containing all sleeve axes.

[0078] Figure 10. Front view of device variant 2 in a configuration where sleeve replacement is permitted. Components such as the pressure beam, sleeve, and O-ring are shown in cross-sectional form in the plane containing all sleeve axes.

[0079] Figure 11. A view of the third variant of the device, showing the device with the pressure beam rotated 180 degrees. In this state, the sleeve can be easily installed or removed. This is a supplementary drawing illustrating the working principle of the pressure beam rotation mechanism.

[0080] Figure 12. View of the third variant of the apparatus during extraction. This figure shows a front view of the apparatus. Components such as the pressure beam, sleeves, and O-rings are shown in cross-sectional form in the plane containing all sleeve axes.

[0081] Example 1.

[0082] In a first non-limiting embodiment, Figure 1a shows a longitudinal vertical cross-sectional view of a sleeve 1 with a circular or elliptical internal orifice. The sleeve is made of polypropylene. The sleeve 1 has an inner diameter of 4.5 mm, an outer diameter of 6.5 mm, and a length of 25 mm. On the outer wall, the sleeve 1 has two protrusions 2 with an outer diameter of 10 mm and a thickness of 1.6 mm along its entire circumference. The purpose of the protrusions is to ensure that the sleeve can be placed stably, easily, and repeatedly in the sleeve holder of the device. At its lower end, the sleeve 1 has a sharp edge 3 along its entire circumference. This edge is formed by two surfaces: one is the surface that constitutes the outer wall surface of the sleeve, and the other is the surface that constitutes the chamfer 4 of the inner bottom of the sleeve. The angle between these two surfaces is 45°. In addition, the sleeve 1 has an orifice 5 with a diameter of 1 mm at its upper part. The orifice on the side wall is used to equalize the internal and external pressures of the sleeve when the inserted pipette fits tightly against the inner wall of the sleeve. Otherwise, when the solution is drawn / drawn from the sleeve, a vacuum will be created, which will prevent all the solution from being extracted. This side hole prevents this effect.

[0083] The sleeve, made of plastic (preferably polypropylene), is cost-effective. This plastic has a degree of flexibility, which makes it easy to conform to the surface of the glass substrate, thus achieving a seal with less pressure.

[0084] If the sleeve is made of acid-resistant steel, greater pressure needs to be applied to the glass carrier. However, if the carrier is made of plastic or aluminum, the steel sleeve can easily seal against the carrier.

[0085] In an alternative embodiment, the sleeve may be made of acid-resistant steel and other plastics such as polyetheretherketone (PEEK).

[0086] Example 2.

[0087] In the second embodiment shown in Figure 2a, the sleeve 1 has a circular or elliptical internal hole, wherein the sleeve 1 has a sharp edge 3 at the lower end of the bottom of the inner wall. The sleeve 1 has a sharp edge 3 along the entire circumference of its outer wall at its lower end. This edge is formed by two surfaces: one is the surface constituting the inner wall surface of the sleeve, and the other is the surface constituting the chamfer 4' at the bottom of the outer side of the sleeve. The sleeve is made of polypropylene. In another variant, it is made of acid-resistant steel, or in yet another variant, it is made of polyetheretherketone (PEEK).

[0088] In the variant shown in Figure 2b, a bottom cross-sectional view of a sleeve with a circular inner bore is displayed. Figure 2b shows the circular inner bore of the sleeve and the circular sharp edge 3 at the bottom of the inner wall. In the alternative variant shown in Figure 2c, a bottom cross-sectional view of a sleeve with an elliptical inner bore is displayed. This figure shows the elliptical inner bore of the sleeve and the elliptical sharp edge 3 at the bottom of the inner wall.

[0089] Example 3.

[0090] In the third sleeve embodiment shown in Figure 3a, the sleeve 1 has a circular or elliptical internal hole, wherein a sharp edge 3 at the bottom of the sleeve 1 is located between the outer wall and the inner wall of the sleeve. This edge is formed by the surfaces of an outer chamfer 4'' and an inner chamfer 4'''. The sleeve is made of polypropylene.

[0091] In the embodiment shown in Figure 3b (which shows a bottom cross-sectional view of a sleeve with a circular inner hole), it can be seen that the inner hole of the sleeve is circular, and there is a circular sharp edge 3 between the outer wall and the inner wall at the bottom of the sleeve 1.

[0092] Figure 3c shows an alternative variant, displaying a bottom cross-sectional view of a sleeve with an elliptical internal bore. The figure shows the elliptical internal bore of the sleeve and the elliptical sharp edge 3 between the outer and inner walls of the bottom of the sleeve 1.

[0093] Example 4.

[0094] Figure 4 shows a sleeve 1 with its sharp edge 3 having a passivation treatment 6 along its entire circumference at the junction of the chamfered surface 4 and the outer surface of the sleeve wall 1 (Figure 5). The passivation width is 0.1 mm. The passivated surface is perpendicular to the outer surface of the sleeve wall and forms a 135° angle with the chamfered surface 4 on the inner side of the sleeve 1. Near the upper end of the sleeve, opposite the lower end with the sharp edge 3, there is a hole 5 with a diameter of 1 mm in the sleeve wall.

[0095] The sleeve 1 may be without a stopper, or it may have a stopper 7 at the hole opposite the sleeve bore with the sharp edge 3. In this variant, the stopper 7 has a hole with a maximum diameter of 2 mm.

[0096] Passivation can also be performed on any type of sleeve described in this application.

[0097] The stopper may also be present in other variations of the sleeve.

[0098] Method Implementation Examples

[0099] In one non-limiting embodiment, Figures 7a-f illustrate a method for extracting substances from a chromatographic plate.

[0100] According to the proposed invention, in the first step, a micro-container is created between the carrier plate of the chromatographic plate and the sleeve pressed on it, wherein the sleeve is pressed at a right angle against the carrier plate of the adsorption layer, such that the bottom is the carrier plate of the adsorption layer, and the wall of the micro-container is the wall of the sleeve. As shown in the figure, in order to create the micro-container, the sleeve 1 is vertically lowered onto the chromatographic plate 8 with the adsorption layer 9 and the glass carrier plate 10 at the position of the adsorption layer 9 where the spot of the substance to be extracted is located (Figure 7a). Arrow 11 indicates the direction in which the sleeve 1 is lowered into the adsorption layer 9 until it contacts the adsorption layer 9, and then the sleeve 1 is pressed vertically downward with a sharp edge and a small pressure as indicated by arrow 12, thereby achieving direct contact and tight connection between the sharp edge 3 of the sleeve 1 and the glass carrier plate 10 (Figure 7b). In this way, a micro-container is formed, the wall of which is the wall of the sleeve 1, and the glass carrier plate 10 constitutes the bottom of the micro-container. The carrier plate 10 with the adsorption layer 9 is referred to as the chromatographic plate 8. The chromatographic plate 8 and the sleeve 1 pressed on it form a micro-container. In a preferred embodiment, the microcontainer has a maximum height of 50 mm and a circular internal opening. The size of the microcontainer can be arbitrary, but its inner diameter is preferably 2 mm to 10 mm, or it can be elliptical, with a minor axis of 2 mm to 6 mm and a major axis of 3 mm to 10 mm. In the next step, a predetermined small volume of extract 13 is introduced into the formed microcontainer through the upper opening of the sleeve. A volume of 20 μL to 500 μL of extract 13 is introduced using a syringe or pipette 14 (Figure 7c). The extract is kept in contact with the adsorption layer 9 in the microcontainer for a predetermined short time (from 1 minute to 10 minutes) (Figure 7d), after which extract 15 (i.e., extract containing the extracted substance) is withdrawn from the microcontainer. The liquid is preferably removed from the microcontainer using a syringe or pipette 14 (Figure 7e). The obtained extract 15 is then transferred to a vial for instrumental analysis. In an alternative embodiment, solid particles are separated by filtration before transfer to a vial. After extracting the extractant 15 from the microcapsule, release the pressure on sleeve 1 as indicated by arrow 12 and lift it above the adsorption layer (Fig. 7f). To extract from the next location in the adsorption layer of the chromatographic plate, use a new sleeve 1, or clean and reuse the used sleeve for the extraction process.

[0101] Method Example 2.

[0102] In a second embodiment of the method, a specific amount of extract 13 is introduced into a microcontainer, and preferably a syringe or pipette 14 is used to extract a portion of it from the microcontainer and immediately reintroduce it into the microcontainer. This extraction and introduction of the liquid is repeated multiple times within short time intervals. The number of extraction and introduction operations is determined experimentally, as is the duration of these extraction and introduction operations.

[0103] As an alternative to performing the above-described multiple extraction and introduction of a portion of the extract, the extract in the micro-container can be stirred.

[0104] Device Example 1.

[0105] Figure 8a schematically illustrates an example apparatus with a sleeve 1 positioned above a chromatographic plate 8 (consisting of an adsorption layer 9 and a carrier plate 10). This apparatus is a mechanism that vertically presses the sleeve 1 against the chromatographic plate, which is placed on a rigid platform 16 of the frame. The main body / frame 17 of the apparatus consists of the platform 16, a support frame 18 attached to the platform 16, and a frame beam 25 attached to the support frame 18. In this embodiment, the sleeve 1, made of polypropylene, is held at its lower edge 3, approximately 10 mm from the adsorption layer 9, by a fastening element in the form of a handle 19. The support 19 of the sleeve 1 is connected to a 5 mm thick flexible pressure pad 20, directly above which is a lower pressure plate 21, connected to an upper pressure plate 23 by turnbuckles 22. An arm 24 of a pressure lever is attached to the upper pressure plate 23. The pressure lever is not shown in the figure; it is not visible because it is located on the back of the apparatus shown in Figure 8a. The lever is attached to the frame beam 25 from the back. Only the lever frame 24 is visible from the front. The lower pressure plate 21 is vertically slidably connected to the frame beam 25, and the movement of the lower pressure plate 21 relative to the frame beam 25 is limited to the distance between the lower limit bar 26 and the upper limit bar 27 attached to the frame beam 25. Two springs 28 are provided between the lower limit bar 26 and the lower pressure plate 21 to assist the upward movement of the lower pressure plate 21, along with the sleeve bracket 19 with the sleeve 1, the turnbuckle 22, and the upper pressure plate 23, when the pressure lever arm 24 is lifted. Two rigid support frames 18 are respectively attached to opposite sides of the rigid worktable 16, and the frame beam 25 is horizontally attached to these two rigid support frames 18 above the rigid worktable 16 (the lower pressure plate 21 and the lever with the pressure arm 24 are slidably vertically attached to the frame beam 25).

[0106] Figure 8b shows an exemplary device, identical to that in Figure 8a, in a state where the sleeve 1 is pressed against the chromatographic plate 8 placed on the rigid worktable 16 of the frame. In the figure, the arm 24 of the pressure lever applies pressure sequentially to the upper pressure plate 23, the turnbuckle 22, the lower pressure plate 21, the elastic pressure pad 20, the sleeve support 19, and the sleeve 1, with the sleeve pressing vertically against the chromatographic plate with its bottom edge. Figure 8bThe deformation of the elastic pressure pad 20 under pressure is shown. Due to the presence of the elastic pressure pad 20, the contact between the lower edge 3 of the sleeve 1 and the glass carrier plate 10 is achieved with the same force across the entire circumference of the sleeve 1 edge 3. The force pressing the sleeve against the chromatographic plate is adjusted by the turnbuckle 22. The lower end of the sleeve 1, which is in direct contact with the chromatographic plate 8, has a sharp edge 3 along its entire circumference, formed by the outer surface of the sleeve 1 and the chamfered surface 4 of the inner sleeve 1. The angle between these surfaces is acute, at 45°. Figure 8c shows an enlarged lower portion of an exemplary sleeve 1 and a chromatographic plate 8 with a visible adsorption layer 9 and a carrier plate 10. The figure shows that the diameter of the sleeve 1 increases slightly at the portion where its edge 3 contacts the carrier plate 10. This results in a slight expansion of the adsorption layer 9, allowing the edge 3 of the sleeve 1 to better conform to the carrier plate 10. This slight increase in the lower diameter of sleeve 1 is achieved by the force applied by the arm of pressure lever 24 and by the shape of the lower part of sleeve 1 given by the chamfered surface 4 and the outer surface of the sleeve 1 wall. In an alternative embodiment, the sleeve is made of acid-resistant steel and has the edges shown in Figures 1-6.

[0107] Device Example 2

[0108] In one non-limiting embodiment, an automated apparatus for extracting substances from an adsorption layer is described, in a four-channel version.

[0109] The apparatus according to the invention in this embodiment is shown in FIG. 9 in the pre-extraction state and FIG. 10 in the state during extraction with the sleeves 1 clamped. It includes four sleeves 1, O-rings 29, a mounting beam 30, and a pressure applying mechanism 31. The apparatus body 32 provides a rigid connection between the pressure applying mechanism 31 and the worktable 33 for placing the chromatographic plate 8. The sleeves 1 are placed in the mounting beam 30, and because an independent O-ring 29 is used for each sleeve 1, its lower edge can freely conform to the surface of the carrier plate 10 of the chromatographic plate 8. The O-rings 29 have a dual function—they connect the sleeves 1 to the beam 30 and hold them above the chromatographic plate 8. The grooves 34 in the mounting beam 30 are used to fix the O-rings 29, and their inner diameter is 30 mm smaller than the diameter of the sleeves 1, which ensures their stable positioning within the beam 30. The chamfering of the upper edge of the sleeves 1 facilitates their insertion into the O-rings 29.

[0110] The pressure application mechanism 31, responsible for the movement of the mounting beam 30 relative to the chromatographic plate 8, can take the form of a pneumatic linear actuator or a screw mechanism driven by an electric motor. This mechanism is additionally equipped with a pair of strain gauge sensors 35 for precise pressure regulation. Once the set force is reached, the control system stops the actuator or motor, maintaining a constant force level throughout the process. The use of a dual pressure application mechanism ensures uniform force distribution across all sleeves 1, which is crucial for obtaining optimal extraction results. In cases where the mounting beam 30 and the chromatographic plate 8 are not perfectly parallel, the O-ring 29 allows the lower edge of the sleeve 1 to flexibly adjust to conform to the surface of the carrier plate 10 of the chromatographic plate 8, compensating for any deformation of the components caused by pressure or component tolerances.

[0111] Alternatively, in addition to O-rings, other flexible elements can be used that allow the beam to connect to the sleeve while maintaining its adjustable function.

[0112] The described scheme can be similar to the described four-channel example and used in devices with more or fewer channels. Due to its modular design, the device can be adapted to various research needs, ensuring high flexibility and efficiency in the extraction process from the adsorption layer.

[0113] Device Example 3

[0114] The apparatus according to the present invention in this embodiment is shown in the accompanying drawings. Figure 11 shows the state for easy replacement of the sleeve—the mounting beam is flipped upwards; Figure 12 shows the state during extraction where the sleeve is pressed against the chromatographic plate.

[0115] In this variant, the chromatographic plate 8 is pressed against a static mounting beam 36 equipped with a sleeve 1 and an O-ring 37. The chromatographic plate is placed on a movable stage 38, which, similar to the second variant, uses a dual pressure mechanism 39, which can take the form of a pneumatic actuator or a screw mechanism driven by an electric motor. The system is equipped with two strain gauge sensors 40 for precise pressure control, maintaining it at a constant level throughout the extraction process.

[0116] The role of the O-rings 37 is crucial here—they act as a connector between the sleeve 1 and the pressure beam 36, while allowing the lower edge of the sleeve 1 to freely conform to the surface of the plate 10 supporting the chromatographic plate 8. This design compensates for any imprecision in the assembly manufacturing process and deformation caused by pressure. The O-rings 37 allow the sleeves 1 to move within the beam 36, so that each of them can be adjusted according to the unevenness of the chromatographic plate 8 or the pressure beam 36, which ensures proper adjustment and optimal contact during extraction. Using O-rings 37 with an inner diameter smaller than the diameter of the sleeve 1 they mate with ensures that they are securely held within the groove 41 of the beam in Figures 11 and 12. The chamfer 42 on the upper edge of the sleeve makes it easier to insert into the O-rings 37. The chamfer on the upper edge is also applicable to other sleeves according to the invention.

[0117] In a key innovation of this variant, the clamping beam 36 is connected to the rotating mechanism 43 (connected to the main body 44), allowing it to rotate 180 degrees. This design enables rapid installation and removal of the sleeve 1. Once the sleeve is loaded via the robotic arm, the beam 36 rotates toward the chromatographic plate 8, and the clamping process then begins.

[0118] Even when the clamping beam 36 and the chromatographic plate 8 are not perfectly parallel, the O-ring 37 allows the sleeve 1 to be flexibly adjusted to compensate for any manufacturing errors or deformations, thereby improving the reliability and efficiency of the device.

[0119] In this example, the sleeve can be changed manually by the operator or by a manipulator arm, which can be part of the extraction unit or operate as a separate device that works in conjunction with the extractor.

[0120] Thanks to the use of the rotating mechanism 43 and the automated robotic arm, the sleeve replacement process is significantly simplified and automated, which improves the efficiency of the device and reduces the time required to prepare for subsequent extraction operations. The use of O-rings and the elasticity of the sleeve ensure proper fit with the chromatographic plate even in the event of slight deformation or unevenness.

[0121] Application Examples

[0122] The following describes an example of the device's application in coccidiostats (substances used as feed additives in farm animals to prevent coccidiosis). Experiments were conducted with five selected coccidiostats: maduramicin, narasin, salinomycin, monensin, lasalocid, and nigericin as an internal standard. Stock solutions were prepared by weighing appropriate amounts of each substance and dissolving them in methanol to a concentration of 1000 µg / ml for each coccidiostat. For solutions in pure solvents, calibration curves were prepared from the stock solutions to achieve final concentrations of coccidiostats in the samples of 0.1, 0.4, 0.9, 2.0, 4.0, 8.0, and 12.0 mg / L. The internal standard concentration was 0.25 mg / ml. For the solution of the substance added to poultry feed, 2.5 g of feed was weighed into a 50 ml polypropylene centrifuge tube, and then a working solution prepared from the stock solution was added to make the final concentrations of coccidiostats in the sample 0.1, 0.4, 0.9, 2.0, 4.0, 8.0, and 12.0 mg / kg, respectively, with an internal standard concentration of 0.25 mg / kg. The sample was then shaken. 10 ml of acetonitrile was added and the sample was shaken vigorously again. The resulting 5 µl volume of each test coccidiostat solution was spotted onto the adsorption layer of a chromatographic plate with silica gel (HPTLC Silica gel 60 F254) using a semi-automatic pipette. The area of ​​the test substance was extracted using the apparatus described according to the invention. To extract the substance from the spotting location on the chromatographic plate, a 50 µl volume of methanol was introduced into a micro-container formed by the chromatographic plate carrier (8) and the sleeve (1) pressed onto the chromatographic plate, with a pressure equivalent to a mass of 15 kg. During extraction, the top opening of the microcapsule was capped. After 5 minutes, the solution was withdrawn from the microcapsule using a 250 µl microsyringe equipped with a needle tip angled at 90° relative to the needle axis and transferred to a 100 µl vial. These solutions were then filtered to remove solid particles from the adsorption layer of the chromatographic plate.The solutions (extracts) obtained in this manner were analyzed using LC-MS (HPLC 1290 Infinity liquid chromatograph equipped with a DAD UV-VIS detector, Agilent, USA; column: Zorbax Eclipse Plus C18, 4.6 x 100 mm; 3.5 μm, Agilent, USA; mobile phase: 94.9% methanol, 5% water and 0.1% formic acid (v / v); triple quadrupole mass spectrometer: 6460 Agilent, USA) to obtain the dependence of the ratio of the peak area of ​​the test substance to the peak area of ​​the internal standard on the concentration of the substance in methanol and feed samples. The obtained dependence is given as a linear equation and the value of the coefficient of determination R2 is as follows. They were obtained at seven different concentrations of the test substance in the range of 0.1 to 12 mg / dm3. For each concentration, the ratio of the peak area of ​​the substance to the peak area of ​​the internal standard was measured three times.

[0123] Results of solutions of coccidiostats dissolved in methanol:

[0124] Results of poultry feed samples with added coccidiostats mixed with methanol:

[0125] Based on the results, it can be concluded that for the extract obtained using the method and apparatus proposed in this invention, the ratio of the peak area of ​​the test substance to the peak area of ​​the internal standard exhibits a very good linear relationship with the change of the concentration of the test substance.

Claims

1. A method for extracting substances from the adsorption layer of a chromatographic plate / from a specific location within the adsorption layer of a chromatographic plate, characterized in that... Includes the following steps: A micro-container is formed between the carrier plate of the chromatographic plate and the sleeve pressed on it, wherein the sleeve is pressed at a right angle to the carrier plate of the adsorption layer, such that the bottom is the carrier plate of the adsorption layer, and the wall of the micro-container is the wall of the sleeve. Preferably, the height of the micro-container is up to 50 mm. • A predetermined small volume of extract is introduced into the resulting micro-container through the upper opening of the sleeve. The volume is greater than 20 μL, preferably between 20 μL and 500 μL. • In a micro-container, maintain contact between the extract and the adsorption layer for a specified short time, ranging from 1 min to 10 min. • Extract the extract (i.e., the extract containing the extracted substance) from the microcapsule. Preferably, the liquid is removed from the microcapsule using a syringe or pipette.

2. The extraction method according to claim 1, characterized in that, In subsequent stages, the extract is sent for instrumental analysis, or the solid particles of the adsorbed layer are separated from the extract before being sent for instrumental analysis.

3. The extraction method according to claim 2, characterized in that, It includes the step of removing solid particles from the adsorbent layer, wherein the solid particles of the adsorbent layer are separated from the extract and the solid particles are removed by filtration or centrifugation before the extract is placed in a vial, or by allowing the solid particles to undergo a sedimentation process after placement in a vial.

4. The method according to claims 1-3, characterized in that, The micro-container has a circular internal opening, the inner diameter of which is preferably 2 mm to 10 mm, or an elliptical internal opening, the minor axis of which is preferably 2 mm to 6 mm and the major axis of which is 3 mm to 10 mm.

5. The extraction method according to claims 1-4, characterized in that, A portion of the introduced extract (11) is extracted from the microcapsule and immediately reintroduced into the microcapsule. This extraction and reintroduction of the portion of extract (11) is repeated 2 to 10 times.

6. The extraction method according to claims 1-4, characterized in that, It is preferable to use a fresh portion of the extract (11) to repeat the extraction process several times and combine the resulting extract solutions, and the number of extraction repetitions is determined experimentally.

7. The extraction method according to claims 1-6, characterized in that, The extract (11) is mixed in a micro-container, preferably using a micro-stirrer or by shaking or sonication.

8. The method for extracting substances from a chromatographic plate according to claims 1-6, characterized in that, The extraction process can take up to 10 minutes.

9. The method for extracting substances from a chromatographic plate according to claims 1-8, characterized in that, During the extraction operation, the upper opening of the sleeve is preferably closed with a plug / cork with an opening (7).

10. An apparatus for extracting substances from a chromatographic plate, comprising a main body and a stage for placing the chromatographic plate, characterized in that... It includes: • An element for securing at least one sleeve (1), in the form of a bracket (19) and / or a fastening beam (30, 36), and connected to the body (17, 32, 44); • The pressure application mechanism (22, 23, 24, 26, 31, 39) connected to the main body of the device (17, 32, 44) ensures that at least one sleeve (1) is pressed vertically onto the chromatographic plate (8) and is capable of removing at least one sleeve (1) from the chromatographic plate (1); • Elastic elements (20, 35, 40) ensure that the edge of the sleeve (1) fits against the chromatographic plate (8).

11. The apparatus for extracting substances from a chromatographic plate according to claim 10, characterized in that, It has at least one replaceable sleeve (1) made of a chemically inert material (preferably plastic) that is sharp at the end of the entire circumference of the lower edge (3) and the sharp end is formed by the connection of two surfaces of the sleeve (i.e., the outer surface of the sleeve (1) and the surface of the chamfer (4) on the inner side of the sleeve), or the edge (3) is formed by two surfaces: one is the surface that forms the inner wall surface of the sleeve and the other is the surface that forms the outer chamfer (4') at the bottom of the sleeve, or the edge is formed by the surfaces of the outer chamfer (4'') and the inner chamfer (4''').

12. The apparatus for extracting substances from a chromatographic plate according to claim 11, characterized in that, The angle between the surfaces that make up the edge (3) is acute, ranging from 15° to 70°.

13. The apparatus for extracting substances from a chromatographic plate according to claims 10-12, characterized in that, The edge formed at the junction of the outer surface of the sleeve (1) and the chamfer (3) surface of the inner sleeve (1) has a passivation treatment (6) on the entire circumference, and the width of the passivation (6) surface is preferably 0.1 to 0.5 mm.

14. The apparatus for extracting substances from a chromatographic plate according to claims 10-13, characterized in that, The sleeve (1) has at least one hole (5) on its wall near the end opposite to the end with the sharp edge (3), the diameter of which is preferably in the range of 0.3 to 1.5 mm.

15. The apparatus for extracting substances from a chromatographic plate according to claims 10-14, characterized in that, The sleeve (1) has at least one longitudinal groove on its inner wall, starting from the end opposite to the end of the sleeve with sharp edge (3), with a length of up to 10 mm, a width of 0.3-1.0 mm, and a depth of 0.3-0.6 mm.

16. The apparatus for extracting substances from a chromatographic plate according to claims 10-15, characterized in that, The internal cross-section of the sleeve (1) is circular, with a diameter preferably between 2 and 10 mm, or the internal cross-section of the sleeve (1) is elliptical, with a minor axis preferably in the range of 2 to 6 mm and a major axis preferably in the range of 3 to 10 mm, or the internal cross-section of the sleeve has a shape that is close to circular or elliptical.

17. The apparatus for extracting substances from a chromatographic plate according to claims 10-16, characterized in that, The length of the sleeve (1) can be up to 50 mm, preferably between 5 mm and 50 mm, and even more preferably between 10 mm and 50 mm.

18. The apparatus for extracting substances from a chromatographic plate according to claims 10-17, characterized in that, The sleeve (1) has one protrusion / flange or two protrusions / flanges (2) on its outer surface around the entire circumference, or has one recess / groove or two grooves / recesses on its entire circumference.

19. The apparatus for extracting substances from a chromatographic plate according to claims 10-18, characterized in that, The sleeve (1) preferably has a plug (7) at the opening opposite to the sleeve opening with a sharp edge (3), and the plug (7) has a hole with a maximum diameter of 2 mm.

20. The apparatus for extracting substances from a chromatographic plate according to claims 10-19, characterized in that, The bracket (19) is equipped with a snap-fit ​​mechanism for the fixed sleeve (1), wherein the bracket (19) is connected to the pressure mechanism via an elastic element (20).

21. The apparatus according to claims 10-20, characterized in that, The sleeve support is an element in the form of a fastening beam (30, 36), preferably equipped with an O-ring (29, 37) for fixing the sleeve (1).

22. The apparatus according to claims 10-21, characterized in that, The component of the pressure-applying mechanism is a turnbuckle (22).

23. The apparatus according to claims 10-22, characterized in that, The pressure mechanism includes a turnbuckle (22) that allows force adjustment, a lever (24), an upper pressure plate (23), and a lower pressure plate (21) connected to an elastic element (preferably a pair of elastic bodies (20)) that connects the pressure mechanism (21, 22, 23, 24) to the support (19) of the sleeve (1).

24. The apparatus according to claims 10-23, characterized in that, The pressure application mechanism (31) is connected to the fastening beam via a strain gauge sensor (35) and ensures that at least one sleeve (1) is displaced by the fastening beam (30) and at least one sleeve (1) is pressed against the chromatographic plate (8), or the pressure application mechanism (39) is also connected to the worktable (38) via a strain gauge sensor (40) and ensures that the worktable (38) with the chromatographic plate (8) is moved and pressed against at least one sleeve (1).

25. The apparatus for extracting substances from a chromatographic plate according to claims 10-25, characterized in that, It includes an additional element that uses a system that controls the operation of the pressure application mechanism (31, 39) based on strain gauge sensor (35) readings to regulate the pressing force.

26. The apparatus according to claims 10-25, characterized in that, The platform (16, 33, 38) is directly connected to the main body, or the platform is connected to the pressure application mechanism via a strain gauge sensor, wherein the pressure application mechanism is connected to the main body.

27. The apparatus according to claims 10-26, characterized in that, The fastening beams (30, 36) are connected to a rotating mechanism (43) that enables the beam / sleeve to rotate 180°.

28. The apparatus according to claims 10-27, characterized in that, The elastic element is a pair of elastic bodies placed in the pressure-applying mechanism, and / or the elastic element is an O-ring that simultaneously fixes the sleeve.

29. The apparatus for extracting substances from a chromatographic plate according to claims 10-28, characterized in that, It includes a rigid platform (16) of a frame; the frame is equipped with two rigid support frames (18) on opposite sides, and a rigid crossbeam (25) of the frame is horizontally attached above the rigid platform; the device includes a bracket (19) for a sleeve (1), the sleeve (1) being interchangeably and vertically attached therein; two elastic washers (20) several millimeters thick are attached from above to the upper left and right sides of the sleeve bracket (19), and a lower pressure plate (21) is attached from above to the elastic washers (20); one end of a turnbuckle (22) is attached from its upper side to the lower pressure plate (21), and the other end of the turnbuckle (22) is attached to an upper pressure plate (23); the upper pressure plate (23) is connected to the arm (24) of a pressure lever attached to the frame crossbeam (25); the lower pressure plate (21) is connected to the frame crossbeam (25) in a manner that allows it to slide vertically; wherein, the lower pressure plate (21) The distance between the upper pressure plate (23) and the turnbuckle (22) is adjusted by shortening / extending the range of the turnbuckle (22); the range of motion of the lower pressure plate (21) is limited between the lower limit bar (26) and the upper limit bar (27) attached to the frame beam; two springs (28) are provided between the lower limit bar (26) and the lower pressure plate (21) to support the upward movement of the lower pressure plate (21) and the sleeve bracket (19), sleeve (1), turnbuckle (22) and upper pressure plate (23) together with it when the pressure lever arm (24) is raised.

Images