HUT60934A - Process, apparatus and sorbent layer for thin-layer chromatography - Google Patents

Abstract

In a method of and an arrangement for carrying out separation analysis investigations in a planar system with forced flow of a fluid carrier medium, an overpressurizable chamber is created for receiving a sorbent layer (4) bearing a complex sample to be investigated and having channels where a fluid carrier medium (eluent) can flow. Within the overpressurizable chamber at least one barrier (7) is applied for interrupting the flow of the fluid carrier medium in the sorbent layer (4) is arranged. The separation unit comprises at least one separation plate having a sorbent layer for ensuring conditions of straight line front migration of a fluid carrier medium, and the at least one separation plate is connected with a closing frame having at least one barrier (7) for interrupting the flow of the fluid carrier medium in the sorbent layer (4) when the closing frame is pressed to the sorbent layer (4).

Description

PROCEDURE, APPARATUS AND SORBENE LAYER FOR RESISTANCE FLUID CHROMATOGRAPHY TESTS

Applicants: Emil Mincsovics ckl ·. wealth engineer, Szentendre, 35% Dr. Ernő Tyihák oki. ve§yészmér-Rek> Budapest, 30% Baranyi Zoltánná ekl. goposman, Budapest, 22% Barnabás Tapa, viH-emosmór ·, Budapest, 13% 8 <2 £.

Budapsgt ^ - 1990. plantsuztuo

FIELD OF THE INVENTION The present invention relates to a method, apparatus and hair field for dimensional, plenum or non-plenum, forced-flow, preferably overpressure, or electric field or individual force-response tests and combinations thereof. According to the proposed method, one-dimensional, linear unidirectional, bi-directional, two-dimensional and three-dimensional, circular, anticircular / triangular / forced-flow developments are carried out in a device designed for this purpose by virtue of its structural, external pneumatic, hydraulic or mechanical pressure or suitable for securing the edges of sorbent wires, also under external pressure, e.g. for covering sheets, closures, or suitable for carrying a sheet cut; sorbent layer (s) without a sheet which are suitably formed at the edges, preferably at the edges. integrated with cover plates, locking frames, to prevent undesired migration of the vehicle / eluent / due to external pressure. flow is prevented, i.e., the carrier can be forced to flow.

In one of the embodiments of the apparatus constructed according to the proposed method, at least one sorbent layer, with or without a carrier sheet, which can be sealed in accordance with the invention, is provided with at least one sample and the spread »• ·

migrating on the liner to different distances specific to the components, and once the carrier has reached any distance, the flow is stopped and at least one separated component is measured on the sorbent layer. Alternatively, the forced-flow carrier may be continuously flowed, and the carrier (s) leaving the edge of the laminate will be continuously captured, and at least one of its constituents will be measured in the flow vehicle. Alternatively, the two process variants can be combined.

The apparatus for carrying out variations of the method comprises a sorbent layer, a carrier unit for delivering to the spreader layer, a unit layer of a sorbent, and a detector with at least one measuring element coupled to the outlet unit. Suitable embodiments of the method variants are also the apparatus variant which enables multiple development and offline evaluation of at least one component, at least one component, of at least one sorbent layer sealed according to one of the methods of the invention. The apparatus variant of our method also comprises using the apparatus for sample purification so that, after removal of interfering substances in the sample, the components to be analyzed are extruded at the edges by one of the methods of the invention.

The present invention also provides an effective elution of 3 sealed sorbent layers, in which case a forced flow is created in a direction perpendicular to or parallel to the positive pressure created by the overpressure.

According to the present invention, the efficiency of high pressure layer chromatography can be significantly increased.

One important type of separation technique is columnar and layer chromatography techniques, in which the migration of the vehicle by capillary force or forced flow results in the disintegration of the samples into components.

Particularly preferred is the use of forced flow for efficient flow of the carrier, which in the case of Layer Liquid Chromatography techniques is by pressure, electro-osmosis, centrifugal force, electric field, vacuum st & effect. An example of the creation of a pressurized flow by pressurization is disclosed in GB 1,570,760, which describes a layer-chromatographic apparatus. The apparatus comprises a layer of sorbent arranged in a pressurized chamber, above which is a pressurized cushion system, for example. over a layer of silica gel, alumina, or other sorbent material, the vapor space is eliminated and pressurized. The chromatographic eluent / carrier is applied to the selected point of the sorbent layer by means of a micropump. At least one side of the laminate is sealed with an impregnation or plastic film and the eluent inlet arrow is used to migrate the eluent on a linear front on the laminate.

- before 4 channels are formed or wire or metal. plastic sheeting is placed.

t

An example of the possibility of further developing overpressure layer chromatography is U.S. Patent No. 4,469,601 to Beaver et al., Which provides an example of two, or even theoretically, three-dimensional separations. After developing in the first dimension, the laminate is dried, so that the eluent is removed from its surface, and then, in the second dimension, a further development is carried out and the amount of components derived from the laminate is quantified. The efficiency of the basic technique can be greatly enhanced by the simultaneous development of more than one layer sheet, preferably by perforating one or more layer sheets on the substrate, preferably at the point of application of the eluent, and by precisely fitting the sheets.

According to the latest development, in the case of pressurized layer chromatography, the constituent streams are measured in separate streams of the carrier leaving the layer edge. The determination of the constituents is continued until a specific retenolation of the components migrated with the nucleus is achieved, then the lamina is moved relative to the chromatographic and / or electrophoretic sensing units and the characteristics of the remaining constituents are measured (Hungarian Patent No. 196,509).

The presented methods and apparatuses have developed the high-performance liquid chromatography techniques of high-performance liquid chromatography, the high-pressure layer chromatography, but the major problem is that the linear one-way and two-way or two-dimensional developments need to seal the edges of the layers. by impregnation so that the pressurized eluent cannot escape from the layer. This wind barrier requires the addition of a manual step, as well as a post-production step, to produce a properly trained plywood board, thereby significantly increasing their cost.

The sealing of the ply sheets also raises another problem, namely the difficulty of producing a wind-impregnating solution that is resistant to a variety of polar and apolar eluents.

A further problem is that organic polymer or other solutions used to impregnate the sorbent layer may contaminate the surface of the layers and, in addition to reducing the efficiency of chromatographic separation, also interfere with the structural tests. they can co-elute with the material to be analyzed. for mass spectrometry.

In addition to contaminating the laminate sheets with the components of the wind-impregnating solutions, it should be noted that prior art sorbent preparation technologies generally use an organic base adhesive to fix the sorbent layer. The different

- 6 ·· ········································································································································································ small and medium molecules of adhesives are equally disturbing in separation but also in structural tests following preparative tests.

In linear one-dimensional development, it is not always possible to separate all the components of the sample, and there may be more than one material in a single spot, which is sometimes the case with two-dimensional development.

Increasing the effectiveness of pressurized layer chromatography also raises the problem of specimen cleanliness. One of the advantages of the technique so far has been that relatively good separation could be achieved without conventional purification, or conventional pre-purification was achieved, which in any case yielded better results than conventional layer chromatography, but this is not enough now; takes advantage of the basic technique for this purpose and, at the same time, based on the similarities between the high performance liquid chromatography techniques of column and layer systems, this operation step creates a special relationship between the two techniques.

SUMMARY OF THE INVENTION The object of the present invention is to solve these problems of pressurized layer chromatography and to meet new needs by providing a method and apparatus meeting the requirements and providing sorbent layers and / or covers and / or frames with or without a solid or elastic carrier sheet.

It has now been found that the edges of sorbent layer sheets can be sealed at any location and size by extruding by hydraulic, mechanical or pneumatic extraction, e.g. sealing the film edges with a film of polymer solution.

It has also been recognized that development efficiency can be enhanced by multiple development on the same layer, and has been found to significantly inhibit the potential for degradation observed in similar developments in the presence of air in a closed system.

One of our new realizations is that the separable complexes can be separated very well into their components in a three-dimensional system. For this, we have developed a practically feasible three-dimensional system.

An important element of our discovery is that for efficient assays, pre-purification of the samples is accomplished using the principle of high performance liquid chromatography, allowing the use of purified samples in high performance liquid chromatography (HPLC) and high performance liquid chromatography (HPLC).

In order to solve this problem, we have developed a procedure and equipment on the one hand and a special sheet and • • · ·

- 8 sheets without liners, inserts and locking frames are formed.

According to the proposed process, linear development methods that are particularly advantageous in pressurized layer chromatography are carried out on sorbent layers without impregnated edges with or without edges, thereby eliminating a significant separation disturbance and impurities of the test substances. In the case of the Layer Sheet thus used, at least one sample and the constituents determined from the sample are delivered in a sequential order which is forced to flow and the constituents of the sample are migrated in separate streams in the sorbent layer and then the flow is terminated at any distance. at least one separated component is measured on the sorbent layer. We can do this \

and to continuously stream the forced-flow carrier, then capture the individual streams of the carrier leaving the edge of the laminate and combine in a material stream and measure at least one of its constituents in the flow. The two solutions can be combined arbitrarily, as was done in the Hungarian Patent No. 169,509.

As a carrier, chromatography eluent and / or solvent is applied to the sorbent layer, the forced-flow components removed from the sorbent layer are measured with a flow cell detector unit, and the residual components on the sorbent layer are measured with a densitometer. The densitometer and detector can be the same unit, both off-line and on-line.

· · · · · · · · · · · · · ·

- 9 A controlled solution is the controlled change of temperature on the surface of the laminate or at a given point on its surface.

The forced flow of the carrier can be provided, inter alia, by pressurization, electro-osmosis, vacuum, or centrifugal force. Chromatography and electrophoresis can be performed simultaneously by applying an electric field in the vicinity of the sheet. Preferably, the electrode differential electrodes are applied to the laminate, in the same direction or perpendicular to the overpressure flow, with the poles optionally changing.

In a pressurized chamber, the base body may be successful / planar / or cylindrical / non-planar / whereas the top body may accordingly be successful / cylindrical. The sorbent layer (s) or sorbent layer system arranged between the two bodies is suitably pressed against the base body and / or the top body.

The edges of the sorbent layer or sorbent layer or of the sorbent layer or sorbent layer system used in the apparatus according to the invention are sealed with extruded surface overpressure to prevent undesirable migration of the eluent / vehicle / vehicle.

The sorbent or non-carrier sorbent layer or sorbent layer or sorbent layer system is mounted or attached.

with integral locking frame 111. systemmed with locking frames and / or insertions or integral cover or covers.

The process, apparatus, and sorbent layer of the present invention provide high performance separation techniques for both layer and column chromatography and combined electrophoresis.

The invention will now be further exemplified. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Referring to the attached drawing. In the drawing it is

Figure 1 shows three different shapes and systems of the pressurized chamber cover body for superficial sealing of the edges of the sorbent layer and of the extruded surface of the edge, and / or of the eluent / carrier with carrier sheet deformation;

b / double sealing achieved by sorbent compaction providing elastic surface pressure;

c / composite elastic surface pressure solution with carrier sheet deformation, a

Figure 2 shows two different shapes and systems of the body of the pressurized chamber for the sorbent layer. ······························································· · «« <

sealing the edges by providing an overpressure of the sorbent layer surface and the peripheral edge of the edge, namely, a profile that is movable with the base plate;

b / power-displacement profile independent of motherboard; the

FIG. 3 is a view showing the base and top body of the pressurized chamber for forming and sealing a thick sorbent layer; the

Figure 4 is a single-sided sorbent layer with a carrier sheet / a /, a double-sided sorbent layer with a carrier sheet / b / and a non-carrier sorbent layer / o /; the

FIG. 5 is a laminated sheet system for tulip run or on-line circular and anticircular forced flow tests; the

Fig. 6 is a cassette for sealing and activating a sorbent layer in a forced flow system; the

Fig. 7 is a schematic layout of a laminate (a) and a laminate topsheet (b) for multiple layer chromatography; the

Fig. 8 is a two layer sheet system with facing sorbent layers and a sealing frame; the

Figure 9 is a four-layered sheet system with unidirectional sorbent layers and sealing frames; the *

Fig. 10 is a three-layered sheet system with integrated frame with ancient carrier plates; the • ·

Figure 11: Layer sheets for on-line forced-flow separation testing, namely / separate vehicle and sample inlet;

b / same vehicle and sample inlet; the

Fig. 12 is a split and / or gradient system providing an isocratic or gradient system. an undivided sorbent field multichannel on-line pressurized layer chromatography system, namely the / eluent / vehicle / inlet point;

b / eluent / vehicle / inlet at multiple points; the

Figure 13 is a schematic diagram of combined on-line and off-line chromatographic and / or electrophoretic separations; the

Figure 14 is a schematic diagram of three-dimensional high pressure layer separation; the

Fig. 15 is an on-line coupled pressurized layer chromatography and column chromatography system.

They occupy a special place in the process of the invention. sorbent layers, which may be prepared in a known manner, but may also have special designs as described below. In particular, in addition to using a sorbent layer, it is also possible to use two or more sorbent layers during development, or these sorbent layers may be incorporated into a system. In carrying out the process of the present invention, a sheet or sorbent layer is provided. sorbent layers or non-carrier solutions may also be used. Particularly preferred for the formation of sorbent layers is the pore size distribution with different specific surface area / pore distribution,

- 13 differentiated silica gels, alumina, magnesium silicate, talc, inorganic materials, while cellulose and its derivatives, resinous powder, polyamide from organic materials. Experience has shown that very good results can be achieved with chemically bound reverse phase sorbents. A preferred solution is the use of alkali chains of different lengths and special chains, for example Si-O-Si-C bonded. Suitable for carrying silica functional groups such as amino, aminocyano, nitro, diol / vicinal hydroxyl, the sorbent layers prepared therefrom may also lead to advantageous separations. Organic or inorganic ion exchange layers and combinations thereof have also produced positive results. We have also found it advantageous to combine ion exchange and inorganic sorbents.

In the case of size exclusion, cross-linked polymer gels and inorganic gels, e.g. large pores of silica gel based mixtures can be used. When separating optically active compounds, chiral * / optically active / sorbent layers are preferred.

Plastic sheet, metal sheet, glass sheet can be used to make the carrier sheet sorbent layers.

The carrier-based sorbent layer may be homogeneous, mixed with several materials or linked together e.g. two different materials with different properties, e.g. the concentrating zopic layer.

The thickness of the sorbent layer is preferably between 0.05 and 5 »0 mm, and the separation task determines the layer thickness used.

• · »« »♦ · · · · · · · · · · · · · · · · · · ·

- 14 Depending on the conditions of separation, sample application is carried out in a strip or patch onto a dry or wet sorbent layer, either in the feeder / eluent stream or independently.

The surface of the sorbent layer is controlled by temperature and spatial distribution according to the separation conditions, thus increasing the efficiency of the separation *

Adhesive application of organic or inorganic materials or sintering with plastic powders or filaments is used for the layered fixing of sorbent particles with or without carrier sheets.

It is also a preferred embodiment of the invention that the pressure transfer seals in the rigid and / or elastic material and / or elastic material and their combinations are used to direct and collect the eluent. for straight-line or circular or segmented channels or. we create channel systems, with or without a filter.

A particularly effective and unique solution of the present invention is the three-dimensional development. The three-dimensional development theoretically imagined so far is made possible, after the two-dimensional development done separately, by "cubing the carrier-free sorbent layer - with unprepared, separated materials", which also contains a plurality of non-carrier, enter service.

- 15 After development in the 3rd dimension, each sorbent layer can be selected separately, and the sites, reactions, and reactions of the separated materials can be evaluated and performed on them.

The third step of three-dimensional development can also be accomplished by conventional TLC development by moving the cube to the eluent, migrating to the sorbent layer block under the action of the eluent capillary forces.

The efficiency enhancement advantage of multiple development can be well utilized in the system of the present invention because air oxygen is excluded, development time is minimal, and this greatly reduces the risk of decomposition. The benefits of multiple development techniques can thus be fully exploited in this system and the operation steps automated.

The sorbent layers may be enclosed in a cassette, with or without a carrier sheet, whereby an eluent guiding channel may be formed in the cover sheet and provided with a closure frame or frames for sealing the sorbent layer. This system can also be used for three-dimensional separations,

An essential element of the present invention is the combination of sample purity and forced flow development, and consequently, the combination of stratified and columnar high performance liquid chromatography. This can be accomplished by attaching the pressurized chamber to the loop of the HPLC injector loop or alternatively operating it. Another solution is to pre-purify the pressurized chamber by on-line delivery and transfer the desired fraction to the column. Separated by development with conventional TLC,

- Of the 16 major fractions, elute from the localized area to the HPLC column via the pressurized chamber. Alternatively, the components eluting with the outlet (s) may be captured and / or applied directly to a layer and / or column, or sprayed onto a sorbent layer, and developed after drying.

The structural analysis and physico-chemical characterization of the constituents of the fractions eluting with one of the methods of the invention can also be carried out by introducing the desired substance (s) into an efficient, sensitive measuring and / or structural assay system. Such measuring systems include: infrared, ultraviolet spectrometer, mass spectrometry, nuclear magnetic resonance spectrometer and others.

The object of the present invention is well illustrated in Figure 1, whereby the edges of the sorbent layer are sealed with three different shapes and systems. Variant a / is a viable solution with a deformation of the carrier sheet whereby the eluent inlet 1 and the sorbent layer 4 on the carrier plate jj are placed between the pressure-transmitting sealing plates £ with the help of the layer-plate positioning frame 8. Extreme surface overpressure is provided by 7 edge profiles. In the case of variant b /, sealing is performed by means of sorption compression, the use of an inert elastic pressure-transfer cover sheet 9, and the complete sealing is ensured by the leak-tight seal 10. The c / variant is a complex, flexible surface pressurized solution with a deformation of a carrier sheet, with the addition of 11 flexible pressure transfer covers. In Figure 2 • «

-17a is provided on the base body of the pressurized chamber, in the case of variant a /, the profile movable with the anvil plate closes, while the eluent outlet 12 has a similar closure system, while the variant b / 13 has a movable leading edge independent of the pressure plate. The latter solution is preferably used after surface sealing. The use of the 2 inlastic elastic pressure transfer covers is a significant factor for effective closure,

Figure 3 illustrates a system for preparative separation, where complete sealing is provided by leakage sealing gaskets 10, and an important element of the system is an eluent control channel 15 provided with a filter element 15 at the eluent inlet.

Figure 4 illustrates that the sorbent layer may be on one side of the carrier sheet portions / a, this is the conventional solution, but it may be the sorbent layer on both carriers o / b /, and significant progress is made on the carrierless solution / c / which e.g. polymerization or ceramic base.

Figure 5 shows a solution for tulip run or on line circular and anticircular overpressure layer chromatography, where the sample application sites 16,

Fig. 6 shows a cassette which is also capable of sealing the eluent guide between the base and the cover plate of the cassette 18, but also showing the eluent collecting channel 17; ···· · ··· <

·· • 4

- 18 na plus 19 locking frames ·

One of the most attractive variants of overpressure layer chromatography is overpressure multilayer chromatography, which uses more than one layer sheet per development, so it can perform the separation of a large number of samples per development. Figure 7 illustrates the conditions for the use of the multilayer chromatography system according to the present invention, whereby the perforation topsheet 21 is a positioning element of this system. Figures 8 and 9 · show two or two figures, respectively. depicts a four-layer sheet system, facing or opposite. with one-way facing sorbent layers and sealing frames. Figure 10 illustrates a three-layered sheet system with integrated 22-frame carrier sheets.

The coupling between Layer and Column Liquid Chromatography techniques is particularly illustrated by the development of forced-flow Layer techniques. THE

Figure 11 illustrates sorbent layers which can be used for on-line pressurized layer chromatography in accordance with the present invention. Two solutions are presented: for a / there is separate eluent and sample inlet, whereas solution b / means for same eluent and sample inlet.

Split or sub-isocratic or gradient systems an undivided sorbent field multi-channel on-line OPLC system is shown in Figure 12, and there are two options: a / eluent introduction at a single point; b / eluensb © leadership at multiple points. The gradient pump 24 delivers solvents from the eluent tanks 25.

·· ··? · ·· ♦ · · 4 · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

- 19 I i {r <

The / version of Figure 13. on-llne OPLC separation and sat I lr shows paralysis with the electrophoretic combination in the direction of development. Detection is solved with 26 deIl (I tectal annulus), while 27 electrodes are located behind or prior to the eluent discharge.

In variant b / a, the electrodes 27 are arranged parallel to the direction of development. Variation c / 27 shows a schematic representation of the use of electrode pairs individually or simultaneously.

Figure 14 is a schematic diagram of three-dimensional OPLC, where sample / 16 represents the start state, while b / shows the result of unidirectional development, 28 is the eluent front, and 29 is the separated material. C / the second dimeh / I nutatja stratallapotot ^one dimensional, while d / shows the separation of the second dimension of the end result. The development in the third dimension e / a is illustrated by a carrier sheet (ff using sorbent layers without ^f , included in the system. F / also shows the eluent control block 30, which is an important element of the smooth elution process.

/ 1 ' ^1, the two-dimensionally developed carrier-free sorbent layer is placed on an embedded layer block without being called up, below the eluent control block 30. In the third dimension

After the conventional or overpressure development I (I ') is carried out, the layers of the sorbent block are separated, and the screen head is written and optionally evaluated.

One sorbent layer of the system, after separation (by s development, either by its own fluorescence or by color), may exhibit an image similar to that shown in Fig. D /.

¹ i

Fig. 15 is an on-line OPLC-HPLC system / illustrated semantically where 3 is the eluent dispenser unit, 32 is the sample feed loop, and 33 is the illustrated HPLC column. 3A is a conventional (eg flow-through) detector system. This system allows the OPLC's first full-HPLC kombinálá1 ¹ <

The use of the same column-switching technique for the use of <RTIgt; lllnt </RTI> so-called solid phase extraction OPLC for sample preparation by HPLC, especially for biological samples.

Claims (28)

Patent claims: A method for performing dimensional, plenum or non-plenum separation tests with forced price and preferably overpressure and / or electric field strength and combinations, wherein at least one sample and a subset of material defined for the sorbent layer or sorbent layer or sorbent layer system are defined. forced flow, continued development of the flow until a specific retention of the components of the sample is achieved, and measurement of at least one of the separated components, We measure that the one-dimensional linear one-way and two-way, two-dimensional self is three-dimensional, cir culinary, anticircular / triangular / propulsion flow improvements are made in a system capable of providing, in addition to pneumatic, hydraulic, mechanical post-sorption-bed overpressure, extruded surface overpressure at appropriate locations to prevent the flow of vehicle in the forced flow. A method according to claim 1, characterized in that the extra surface overpressure is produced at certain locations in the sorbent layer, preferably by a leading edge profile of the base and / or top body of the pressurized chamber. The method according to claims 1 and 2, characterized in that the extrudate overpressure is provided with a preferably sharp profile on the surface of the sorbent layer or on the carrier sheet. 4 / 1-3. A method according to claim 1, wherein the excess surface pressure on the sorbent layer is formed by the base or top body of the pressurized chamber and the sorbent layer or overlay. preferably with a sorbent compacting frame or between the hoioseozing plate. frames along one or more edges. 5 / 1-4. The method of claim 1, wherein the edge-line closure is formed by deformation of the carrier sheet along the edge penetrating the solids bed. 6 / 1-5. The process of claim 1, wherein the extrudate overpressure is applied to the lower and / or upper surface of the sorbent layer. 7 · / Figures 1-6. The method of claim 1, wherein the extra surface pressure is resolved depending on the surface pressure and / or independently. 8. / Apparatus for dimensional, planar, or non-planar, separation flow, preferably at least one sorbent layer, , a unit for applying a carrier to the surface of the sorbent layer, and a fluid conduit to the edge of the sorbent layer, the conduit? ¹ comprising a detector with at least one measuring member and a detecting unit disposed relative to the surface of the sorbent layer relative to one another, characterized in that it extends or extends along the edges of the sorbent layer or sorbent layers, under temperature-programmed conditions. 9. Apparatus according to claim Q, wherein an anti-leak seal is also used to completely prevent the elution leakage. 10. / A 8-9. Device according to Claim 1, characterized in that the pressure-transmitting sealing plates are made of rigid and / or elastic material and / or elastic material or combinations. 11. / A 8-10. Apparatus according to Claim 1, characterized in that the pressure-transmitting sealing plates are used for the orientation and collection of the eluent. straight-line or circular or segmented channels or. we create channel systems, with or without a filter. Apparatus according to claim 8, characterized in that in the circular and anticircular forced flow tests, the eluent inlet or outlet is used. -displacement is solved in one point or in an arc. Device according to claim 8, characterized in that the pressurized chamber is connected to the loop of the HPLC injector loop or alternatively operated. 14./ 8 and I3. Apparatus according to claim 1, characterized in that in the pressurized chamber pre-purification of the on-line is performed and the desired fraction is applied to the column. Apparatus according to claims 13 and 14, characterized in that the main fractions separated by conventional TLC development are eluted from the localized area through the pressurized chamber. Apparatus according to claim 8, characterized in that the components eluting with the outlet (s) in the pressurized chamber are captured and / or applied directly to a layer and / or column and / or sprayed onto a sorbent layer. Apparatus according to Claim 8, characterized in that electrodes which create an electrical potential difference are fitted to the sorbent layer (s). Apparatus according to claim Q, characterized in that the upper and lower bodies of the pressurized chamber are cylindrical, with sorbent cores formed with a curved surface, with or without a carrier plate. 19. Apparatus according to claim 8, characterized in that the fraction containing at least one component obtained in the pressurized chamber according to one of the methods according to the invention is discharged or discharged. outputs to other efficient measuring and / or structural inspection systems. -2 ^ - 20. / A sorbent layer for forced-flow separation studies where a straight-line migration of the eluent is provided by a channel or other eluent guide formed in the sorbent, characterized in that it extends over and over one or two layers of sorbent with or without / or integrated with the carrier plate (s) · as required. An embodiment of the sorbent layer according to claim 21, characterized in that the sorbent layer (s) with or without carrier sheet and the inert elastic pressure-transfer cover sheet (s) form a system or are connected thereto. The sorbent layer according to claims 22 and 21, characterized in that the extrudate overpressure is formed on the sorbent layer on two, three or four sides in a straight line or along a circle delimiting the entire sorbent field or portions thereof. 23. / a 20-22. A sorbent layer according to any one of claims 1 to 3, characterized in that a sorbent layer without a carrier sheet or a sorbent layer on a carrier sheet or two sorbent layers on both sides of the carrier sheet is used. 24 / a 20-23. A sorbent layer according to any one of claims 1 to 3, characterized in that the carrier sheet for the sorbent layer according to the invention is made of glass, plastic or metal. 26 ~ 25. / a 20-24. Sorbent layer according to any one of claims 1 to 3, characterized in that the material of the sorbent layer is silica gel, polar and / or apolar modified silica gel, alumina moxide, talc, magnesium silicate, hydroxyapatite, cellulose, modified cellulose, organic or inorganic ion exchange,? optically active and other organic and inorganic sorbents. 26. / A 20-25. A sorbent layer according to any one of claims 1 to 4, characterized in that the layer system for fixing the sorbent particles with or without the carrier sheet is glued with an organic or inorganic material or with a color powder with a plastic powder or fiber. A sorbent layer according to claim 20, characterized in that the cover sheets, which are placed or assembled to cover the sorbent layer, are made of rigid and / or elastic material and / or elastic material. are made of combinations. The sorbent layer according to claim 20, characterized in that the sorbent layers without carrier sheet are superimposed and provided with an eluent guide block for three-dimensional development. 29. / The sorbent layer of claim 28, wherein the development in the third dimension utilizes conventional TLG development.