RU2216018C2 - Method to realize flow thin-layer chromatography and device for its implementation - Google Patents

Abstract

FIELD: analytical chemistry. SUBSTANCE: method includes preparation of chromatographic chamber, plate, eluent and specimen, continuous supply of eluent on to plate and removal of eluent, deposition of specimen, execution of chromatographic analysis and uninterrupted detection of components of specimen in wetted layer of adsorbent. Chromatographic chamber has cylindrical form and is equipped with motor. Chamber is installed at angle of 0-45 degrees. Standard chromatographic plate is anchored inside chamber on mobile platform and is rotated with speed of 500-3000 rpm. Eluent is continuously supplied into center of plate and spent eluent ( fractions of eluate ) is directed into dish of detector. Specimen with volume from 2.5 microliter to 1.5 milliliter is deposited in center of plate or at point 10-30 mm away from center, circular chromatography and uninterrupted detection are conducted at distance of 35-80 mm from start. Device for implementation of method comprises metal or polymer light-protected chromatographic chamber, chromatographic plate- substrate made of glass, metal or polymer with layer of adsorbent deposited on it put on support and placed inside chamber, unit to supply and remove eluent ( eluate ), light source with light filter and photodetector. Device is distinguished by cylindrical form of chamber that has diameter of 310 mm and height 180 mm. It comes in the form of tray with separable cover. Tray has groove in its bottom located along wall in which eluate is collected and removed outside and into dish of detector over branch pipe. Cover carries cock-measuring box and metering injector placed 10-30 mm away from it. Light source and light filter are built into cover and photodetector is mounted for movement above plate. Mobile metal platform, diameter 290 mm, carrying chromatographic plate standard by size with layer of adsorbent 100-5000 mcm thick and particle size 3-30 mcm is fixed horizontally inside chamber, in center of base of tray on pin connected to motor. Frame of motor of device is solidly joined to chamber from beneath. Motor with chamber is put on support with the aid of screw mounting enabling position of device to be varied at angle of 0-45 degrees. Vessel with eluent, pump, second photodetector, dish and second light source are located outside of chamber. EFFECT: development of method of conducting quick flow thin-layer chromatography with high resolution, mechanization of method. 4 cl, 1 dwg

Description

 The invention relates to analytical chemistry, more specifically to flow thin layer chromatography (PTLC) and a device for its implementation.

 The invention can find application in analytical laboratories that control the production and quality of products, and in medical laboratories in identifying and isolating individual substances with close or small mobility coefficients from mixtures, when determining trace amounts of substances in mixtures.

The following concepts and abbreviations are used in the description:
CHROMATOGRAPHY - a method of separation and analysis of mixtures, based on the different distribution of their components between two phases - fixed and mobile (eluent) [Big Encyclopedic Dictionary. - 2nd ed. - M .: "Big Russian Encyclopedia", St. Petersburg: "Norint", 1997. - S.1323].

 ADSORPTION CHROMATOGRAPHY - chromatography based on the different ability of the components of the mixture to be adsorbed on a stationary phase [Analytical chromatography / K.I. Sakodynsky et al. - M.: Chemistry, 1993. - S. 342].

 LIQUID CHROMATOGRAPHY (LC) - chromatography in which liquid eluent is used [ibid., P.225].

 HIGH-PERFORMANCE LC (HPLC) is an adsorption LC in which a chromatographic column filled with an adsorbent is used as the stationary phase, and the eluent moves under an externally applied high pressure [ibid, p. 225].

 THIN-LAYER CHROMATOGRAPHY (TLC) - adsorption LC carried out on plates coated with an adsorbent layer. The separation of the components of the mixture occurs on an open adsorbent layer, so that three types of elution can be performed: linear, circular, anti-circular. "Planar" version of chromatography. The movement of the eluent occurs under the action of capillary forces. TLC includes the following stages: sample preparation, plate preparation, sample preparation, chromatographic chamber preparation, chromatography (development, chromatogram development), eluent removal from the plate, detection (visualization), identification, quantification (in quantitative analysis) and documentation [there same with 345].

R _f = L _s / L _o (<1) - mobility coefficient;
L _s is the distance from the start line (the place of application of the sample) to the center of the spot on the plate;
L _o - the distance from the start line to the front of the eluent [Osterman L.A. Chromatography of proteins and nucleic acids. - M .: Nauka, 1985. - P.57].

TLC is used as analytical and preparative methods, which provide separation in the range from 10 ^-3 to 10 ^-12 g.

 From the point of view of hardware design and the method of implementing TLC, in contrast to HPLC, the chromatography method is simple and affordable for the mass consumer when identifying substances, that is, in a qualitative analysis. However, these advantages of TLC are lost in quantitative analysis. Quantitative determinations are performed in TLC only at the end of the development of the chromatogram using special precision scanning equipment. In addition, the nonequilibrium nature of the TLC process, combined with the need for each subsequent analysis on a new sorption layer (new plate) adversely affects the accuracy and reproducibility of the results of quantitative analysis.

 Carrying out during the analysis of continuous detection of the components of the mixture in the eluent stream, widely used in HPLC, is not used in TLC. One of the reasons for this is the inconsistency of the mobile phase velocity in TLC, since it is precisely the high accuracy of maintaining the flow of eluent in HPLC column systems that allows us to judge the amount of substance in the chromatographic zone by the peak area in the chromatogram. The only type of TLC, characterized by the constancy of the speed of the mobile phase is PTSH. It is carried out in such a way that the eluent is continuously removed from the end of the flat layer of the sorbent (due to natural evaporation or evaporation when the edge of the plate is heated), while its speed is determined by the capillary properties of the layer and, when the equilibrium is established, the velocity value becomes constant.

 High linear velocity of the eluent and its constancy are achieved in HPLC using precision high-pressure pumping systems. In PTSH, the constancy of the linear velocity of the mobile phase is due to purely capillary effects and requires only evaporation of the solvent to maintain it. However, these same factors determine lower values of the speed of the eluent in PTLC than in HPLC. A well-known attempt to increase the speed of TLC by applying external pressure (by analogy with HPLC) using the Chrompres instrument (Hungary) on plates with a hardened adsorbent layer is not applicable to PTLC because of the technical complexity of implementation [Analytical chromatography / K.I. Sakodynsky et al. - M.: Chemistry, 1993. - S. 362].

 Another obstacle to continuous detection in TLC and in PTLC, in particular, is that all known detectors are adapted to work with transparent solutions (in the eluent stream), and not with light-scattering porous matrices (chromatographic plates), where photometric the law of Lambert - Bouguer - Bera. This problem is solved with some limitations in PTLC when detecting directly in a wetted adsorbent layer on a transparent glass substrate [A. Vorontsov. Flow thin-layer chromatography (PTLC) in the analysis of ecotoxicants: features of the method and criteria for evaluating the results // J. Ecologist, Chemistry, - 1992. - 2. - P. 33-43].

 PTLC with continuous detection of the substance directly in the wetted adsorbent layer is considered an independent method of quantitative chromatographic analysis, which occupies an intermediate position between TLC and HPLC [Analytical chromatography / K.I. Sakodynsky et al. - M.: Chemistry, 1993. - S. 357]. This modification of PTSH is superior to TLC due to the simplification of the quantitative analysis process, however, it is inferior to HPLC in the speed and resolution of the analysis, and the accuracy of the results.

 There is a method of conducting PTX in the analysis of ecotoxicants (chelates of heavy metals, polycyclic aromatic hydrocarbons), which consists in preparing a sample, preparing a chromatographic chamber, preparing a plate and supplying eluent to its edge while ensuring continuous removal of spent eluent from the opposite end of the plate due to evaporation, application samples on the plate to the starting line from the same edge of the plate where the eluent is fed into the eluent stream established in the equilibrium system, chromatograph Studies, continuous detection - identification and quantitative determination of sample components during elution in a wetted layer [A. Vorontsov M. Flow thin-layer chromatography (PTLC) in the analysis of ecotoxicants: method features and evaluation criteria // J. Ecologist. Chemistry. - 1992. - 2. - P.33-43]. The time for one analysis is from 45 minutes to one and a half hours.

 As a device for implementing the known method, a PTX chromatograph is proposed, including a rectangular metal light-shielding chamber with openings for two optical fibers and for air outlet and with a hinged lid, on which a light source is fixed - a DRK-120 spectral lamp and an interference filter; inside the chamber - a container with an eluent; an eluent supply wick from the left edge of the chromatographic plate immersed at one end in a container with an eluent and the other end touching the chromatographic plate; the chromatographic plate itself of a non-standard size of 70x25 mm, consisting of a substrate (glass or aluminum foil) with an adsorbent layer deposited on it (silica gel) and mounted motionless and horizontally; cover plate; a glass plate located above the cover plate with an opening for introducing a sample; an optical fiber holder with a slit for illuminating the sorbent attached above the cover plate closer to its end and to the right of the glass plate so that detection occurs at a distance of 40 mm from the start line; a wick for evaporation of the eluent from the right edge of the chromatographic plate; under the wick, a fitting for blowing it; outside the camera there is a photodetector (photomultiplier tube PMT-95) and a rotary disk with output ends of the optical fibers. The well-known PTX chromatograph allows photoabsorption and fluorimetric detection. This is achieved through the use of flexible fiber optic fibers.

 Compared with quantitative TLC based on the use of scanning facilities, the known method carried out on a PTX chromatograph has several advantages: reproducibility of results increases 6-8 times; densitometry or faorimetric detection in the wetted layer compared to optical scanning, ceteris paribus, passes with a large signal to noise ratio (signal intensity is higher), which is ensured by increased transparency of the wetted layer; in the PTX mode, the light probe is fixed on one section of the adsorbent layer, which removes the contribution of the layer heterogeneity along the length; it is possible to enter a sample on the same plate up to several hundred times, similar to how a sample is repeatedly introduced into a chromatographic column, which further increases the reproducibility of the analysis and leads to significant material savings; analysis of volatile components is possible, which is not allowed by TLC.

 The disadvantage of this method is the requirement to minimize the volume of the injected sample and the impossibility of introducing a larger sample during evaporation of the solvent, as is the practice in TLC when applying the sample to a dry layer in order to concentrate it.

 According to the sensitivity of the determination of a substance, the known method is close to modern HPLC, however, it is significantly inferior to it in the speed of analysis and in resolution.

 The disadvantages of the known method of conducting PTX are largely associated with the disadvantages and limitations of the PTX chromatograph used for its implementation. The movement of the eluent in PTSH occurs slowly, only due to the nature of the adsorbent and the particle size of the capillary forces. Despite the fact that adsorbents with a large particle size (20-25 μm) are used in the known chromatograph, the analysis time is at least 45 minutes. In this case, there is some blurring of the chromatographic zones and a loss of selectivity for a number of substances. Lengthening the sorbent layer over 70 mm (and therefore transferring the detection to a distance of more than 40 mm from the start line) in order to increase the resolution is impractical - the analysis time is too long, the contribution of molecular diffusion to the erosion of chromatographic zones is increased.

 Thus, the problem of improving the PTSH, attractive from the point of view of simplicity and accessibility of instrumentation of the chromatography method, remains relevant.

 The task of the invention is the creation of the express method of PTSH with high resolution, which could compete with HPLC.

 This problem is solved by the proposed method of conducting PTSH and a device for its implementation.

The inventive method of conducting PTSH has the following set of essential features:
1. Prepare a sample of a mixture of shared components.

2. Prepare a chromatographic chamber of cylindrical shape and fix at an angle of 0-45 ^about .

 3. Prepare a standard chromatographic plate with a continuous layer of adsorbent.

 4. Fasten the plate to the movable platform.

 5. Prepare the eluent.

 6. Start to rotate the movable platform with the plate at a speed of 500-3000 rpm

 7. The eluent is pumped to the center of the plate at a rate of 0.10-3.5 ml / min; in this case, gradient chromatography is possible: supply of an eluent of a composition that varies over time; the spent eluent, and then the eluate fractions flowing to the bottom of the chamber pan due to centrifugal force, are sent to the detector cell.

 8. Maintain the system for 1-3 minutes to establish a steady flow of eluent.

 9. A 2.5 μl-1.5 ml sample is applied to the plate; possible application of the sample before the rotation of the plate.

 9.1. In the center of the plate using a metering tap.

 9.2. At a distance of 10-30 mm from the center of the plate using a syringe dispenser.

 10. Chromatography is performed to ensure continuous collection of eluate fractions and their entry into the detector cuvette.

 11. Conduct continuous detection (identification and quantification) of the separated components of the mixture in the wetted layer during chromatography at a distance of 35-80 mm from the start.

 Chromatography occurs within 1.5-15 minutes.

The inventive device for the implementation of PTSH has the following set of essential features:
1. The device includes a metal or polymer light-proof chromatographic chamber of a cylindrical shape with a diameter of 290-310 mm and a height of 60-180 mm, consisting of a tray and a removable cover. There are holes in the pallet: on the wall - for introducing inert gas, on the bottom in the center with a flange - for the fastening pin of the movable platform, and a nozzle, the pallet has a groove along the wall into which the eluate is collected and brought out through the nozzle, to the detector cuvette, and recesses on the wall for installing the cover plate. A multi-way metering valve connected to the pump is fixed on the lid in the center, and 10-30 mm from it - the dosing syringe, the nozzle of the dosing valve and the needle of the dosing syringe pass through the special holes in the lid; a light source and a light filter, a photodetector or a light guide with a photodetector are integrated into the lid, with the possibility of moving it and installing it above the chromatographic plate for detection at a distance of 35-80 mm from the start due to special holes or using a retractable frame made of corrugated material.

2. The device is equipped with a motor. The motor housing is tightly connected to the chromatographic chamber from below; the motor together with the camera is mounted on a support using a screw mount, which allows you to vary the position of the device at an angle of 0-45 ^o . Outside the chromatographic chamber there is a vessel with an eluent, a pump connected to a vessel with an eluent, a second cell photodetector and a second light source.

 3. Inside the chamber in the center at a distance of 5-50 mm from the base of the pallet, a horizontally movable metal platform with a diameter of 285-290 mm with a standard chromatographic plate of 100-200x100-200 mm in size, which is mounted on it, is fixed on a fastener pin connected to the motor a transparent (glass) or opaque (metal or polymer) substrate with an adsorbent layer deposited on it, with a thickness of 100-5000 μm and a particle size of 3-30 μm, the chromatographic plate on top is covered with a UV transparent coating plate Therians (silica, polytetrafluorethylene) with two openings for the spout-tap dispenser needle and a dosing syringe.

 The inventive device allows continuous photometric and fluorimetric detection in a wetted layer. In the case when the material of the cover plate does not pass UV, a window of 30x30 mm is provided in it opposite the light source.

 The specified set of essential features of the proposed method and device provides a technical result - improving the resolution of PTSH by optimizing the conditions of chromatography. Achieved technical result allows us to characterize the inventive method PTSH as an express method (analysis time is 1.5-15 minutes), which has a resolution at the level of HPLC. An additional technical result: the claimed express method of PTSX is more universal in terms of the possibility of analyzing a variety of mixtures compared to analogues, since it allows the use of different adsorbents with any particle size and plate with a wide range of layer thicknesses; to implement the method and obtain good separation, the length of a standard plate is sufficient; high requirements are not imposed on the quality of the layer (as in TLC under pressure), it is possible to use viscous and low-volatile eluents, samples up to 1.5 ml (in the prototype up to 25 μl). In addition, using the proposed method PTSX possible quantitative separation of the components of the mixture as a result of collecting the eluate.

 The proposed PTSH method, including a method of conducting PTSH and a device for its implementation, is simple and affordable in terms of hardware design.

 Distinctive essential features of the proposed method from the prototype method are signs 2, 4, 6-11, describing the operation of fixing the device, feeding the eluent, feeding the sample, performing chromatography using rotation of a chromatographic plate, continuous detection.

 Distinctive essential features of the claimed device from the prototype device are the signs describing the shape and components of the chromatographic chamber and its dimensions, the presence of a motor, its connection with the camera, mounting the device with the possibility of its horizontal and inclined position, solvent input and samples in the center of the device through a tap - and syringe dispensers, the withdrawal of the eluate through the groove of the pallet through the pipe into the detector cuvette, mounting the chromatographic plate on a movable platform connected to the motor, type pl astins - standard in size and characteristics of the adsorbent (particle size, wide variations in nature - different types of silica gels, aluminum oxide, etc.), adsorbent layer, mounting of the cover plate, a movable photodetector with a light guide, the presence of a pump and the location of the pump and vessel with the eluent outside the chamber , the presence and location of the second photodetector and the second light source.

 The analysis of the prior art did not allow to find a solution that completely coincides in the totality of essential features with the claimed method of conducting PTSH and a device for its implementation.

 Known rotational TLC (RTSX), in which the movement of the eluent is due to centrifugal force. For its implementation, devices of the Rotachrome type have been developed, the design of which includes a rotor with a rotating platform (rotation speed 1000 rpm). However, PTX, unlike PTLC, is a variant of traditional TLC with a one-time sample injection and the use of precision scanning devices. The method allows you to work only with a circular version of TLC and is structurally designed for original plates with adsorbent deposited in the form of a ring.

 The disadvantage of this method is that the development of the chromatogram is stretched to a distance of 10 cm, that is, to the edge of the plate. The known method is not used for quantitative analysis. It is still believed that the PTX is not adapted for continuous detection, since it is practically impossible to install the detector in the PTX device [Mazurek M., Witkiewiczh. Rotation planar chromatography // Chem. Anal. (Warsaw). - 1998. - 43.- P.536].

 Thus, the claimed invention meets the criterion of "novelty."

 Only a set of essential features of the proposed method for conducting PTSH and a device for its implementation allows to achieve the above effect. From the fame of certain features of the claimed invention does not follow clearly the possibility of developing a new express TLC method, competing in resolution, sensitivity and time of analysis with HPLC. In addition, the claimed invention solved the complex technical problem of installing the detector in the process of flow-rotation TLC, which until now was considered impossible. Therefore, it was impossible to assume in advance that the application of centrifugal force to the PTSH would be installed in the system and that this would lead to a significant acceleration of the process and a fundamental improvement in the quality of the analysis. Thus, in the claimed invention, a new functional dependence "operation - property" is implemented. This allows us to assert that the proposed invention meets the eligibility condition "inventive step" ("non-obviousness).

 Both proposed objects, method and device, are united by a common inventive concept aimed at solving the problem.

 To confirm the compliance of the decision with the condition of protection "industrial applicability" and to better understand the essence of the invention, we give examples of specific performance.

 The drawing shows a diagram of the inventive device. 1 - chamber pan, 2 - removable cover, 3 - light source with a light filter, 4 - movable photodetector, 5 - multi-way metering valve, 6 - syringe metering device, 7 - inert gas inlet hole, 8 - chromatographic plate mounting pin 9 - a movable platform, 10 - a chromatographic plate, 11 - a cover plate, 12 - a pump, 13 - a vessel with an eluent, 14 - a motor, 15 - a pin-fixture of a moving platform, 16 - a motor support, 17 - a second photodetector, 18 - the second light source, 19 - a cuvette.

 The light source is a DRK-120 spectral lamp, an interference optical filter, a photodetector - an FEU-95 photoelectronic multiplier, a photodiode, and a video camera. Detectors (the base is a photodetector, light source, cuvette) are photometric or fluorometric, for example, a photometric detector of the SKB RAS "Analytical Instrument Making".

 In the invention, standard size chromatographic plates with a layer of silica gel, alumina, modified silica gel with a thickness of up to 5000 μm from Merck (Germany) or domestic, as well as plates prepared as follows, were used.

19 g of KSKG silica gel with different average particle sizes (3-30 μm), 1 g of soluble starch and 36 ml of distilled water are heated with stirring to 85 ^o C, until the mixture thickens. After cooling, a little water is added to adjust the viscosity of the mixture, after which a homogenized suspension is applied to a glass (metal, polymer) substrate, dried in air for 2 hours at room temperature, then activated at 110 ^° C for 40 minutes. Layer thickness after drying 200 microns.

 All solvents (isooctane - eluent, hexane - solvent for the test mixture) were qualified chemically pure. and dried before use with 5A molecular sieves.

For analysis, mixtures of fullerenes C ₆₀ and C ₇₀ from the company Fullerene Technologies LLC (St. Petersburg) were used.

The resolution of Rs for fullerenes is calculated by the following formula [Current state of liquid chromatography. Edited by J. Kirkland. M .: Mir, 1974]:
R _s = 2 (t ₁ -t ₂ ) / (w ₁ + w ₂ ), where w is the width of the chromatographic strip, t is the retention time of the substance.

Samples are prepared as follows:
a mixture of fullerenes is dissolved in hexane. The composition of the mixtures is C ₆₀ : C ₇₀ : 1: 1.1: 2.1: 3.

 Example 1

 An example was carried out in the conditions of the prototype method on a device for implementing the prototype method (PTX). The plate is prepared according to the method above, with silica gel with a particle size of 4 μm. The sample volume is 2.5 μl. The resolution for a 1: 1 fullerene mixture is 0.7. The experiment time is 2 hours

 Example 2

 An example was carried out under the conditions of example 1, the particle size of the silica gel is 20 μm. Resolution is 1.1. The experiment time is 1.5 hours

 Example 3

 An example was carried out under HPLC conditions on a Hypersil ODS column measuring 25 cm x 2.1 mm (inner diameter). Resolution is 6.5. The experiment time is 10 minutes

 Example 4

 An example is carried out in the conditions of the proposed method and on the device for its implementation.

The device is tilted by 35 ^o to a horizontal surface. The plate was obtained according to the above method with silica gel with a particle size of 4 μm. Clear fluoroplastic cover plate.

A sample of fullerenes of composition 1: 1 with a volume of 25 μl was prepared. A chromatographic chamber is prepared. Tilt her. Fix the chromatographic plate on a movable platform. Prepare the eluent - isooctane. They begin to rotate the moving platform at a speed of 500 rpm. The eluent is pumped at a rate of 0.1 ml / min through a metering valve to the center of the plate. Stand for 2 minutes to establish a steady flow of eluent. The metering tap is switched, a sample is introduced into the center of the plate. Switch the dispensing tap to pass the eluent. It is chromatographed. At the same time, at a distance of 50 mm from the start, continuous photometric detection, collection of eluate fractions and quantitative measurements are carried out. The experiment time is 10 minutes The resolution is 1.8. Quantitative analysis results:
Test composition: 1: 1.

 The result obtained: 1.05: 0.96.

 Example 5

 The example is carried out under the conditions of example 4. A plate with silica gel with a particle size of 20 microns. Speed of rotation is 500 rpm. The resolution is 2. The experiment time is 15 minutes.

 Example 6

 The example is carried out under the conditions of example 4. A plate with silica gel with a particle size of 4 μm. Rotational speed 2000 rpm. The resolution is 4. The experiment time is 10 minutes.

 Example 7

 The example is carried out under the conditions of example 4. A plate with silica gel with a particle size of 20 microns. Rotational speed 2000 rpm. Resolution is 1.4. The experiment time is 10 minutes

 Example 8

 The example is carried out under the conditions of example 4. A plate with silica gel with a particle size of 4 μm. Rotational speed 3000 rpm. Resolution is 3.5. The experiment time is 10 minutes

 Example 9

 The example is carried out under the conditions of example 4. A plate with silica gel with a particle size of 10 μm. Rotational speed 2000 rpm. The resolution is 5. The experiment time is 10 minutes.

 Example 10

 The example is carried out under the conditions of example 4. A plate with silica gel with a particle size of 30 μm. Rotational speed 2000 rpm. Resolution is 2.2. The experiment time is 10 minutes

 Example 11

 The example is carried out under the conditions of example 4. The thickness of the silica gel layer on the plate is 100 μm. Resolution is 2.0. The experiment time is 10 minutes

 Example 12

 An example is carried out under the conditions of example 4. The installation is horizontal. Resolution is 1.7. The experiment time is 15 minutes

 Example 13

 The example is carried out under the conditions of example 4. Detection is carried out at a distance of 40 mm from the start. Resolution is 1.7. The experiment time is 10 minutes

 Example 14

The example is carried out under the conditions of example 4 on a plate RP ₁₈ firm Merck at a speed of rotation of 2000 rpm The sample volume is 50 μl. Resolution is 6.5. The experiment time is 15 minutes

 Example 15

 The example is carried out under the conditions of example 2 on a Merck plate. Resolution is 1.3. The experiment time is 1.5 hours

 If necessary, thermostating of the process is possible.

 The claimed invention is not limited to the above examples.

 The data of the examples indicate that as a result of the invention, a new PTSX method was obtained - flow-rotational (RPTSX), characterized by expressivity and high resolution. Compared to the prototype method and apparatus, the time of quantitative analysis decreased by 4-10 times and approached the time of HPLC (2-10 min), the resolution exceeds the resolution of PTLC by 5-6 times and reaches the lower limit of HPLC. The invention solved the complex technical problem of the compatibility of flow processes, the application of centrifugal force and continuous detection. Thus, the claimed invention solved the task of creating a simple in hardware design competing with HPLC TLC method.

 The claimed invention allows the use of the same plate repeatedly, as well as continuous multiple analysis (chromatography - washing the plate with eluent - the next chromatography without replacing the plate).

 Exceeding the declared interval parameters makes it impossible to implement the invention: the resolution of the method decreases. When the rotation speed increases above 3000 rpm, the eluent flows along the surface of the plate.

Claims (4)

1. The method of conducting thin-layer chromatography, including the preparation of a chromatographic chamber, a chromatographic plate consisting of a substrate and a layer of adsorbent, eluent, sample deposited on it, continuously supplying the eluent to the plate and removing the eluate from the plate, applying the sample to the plate, chromatography and simultaneous continuous detection of sample components in a wetted adsorbent layer, characterized in that a cylindrical chromatographic chamber equipped with a motor is used in preparing the chromatographic chamber is installed at an angle of 0-45 ^o , the standard chromatographic plate is fixed inside the chamber on a movable platform and rotated with a motor at a speed of 500-3000 rpm. / min, the eluent is continuously fed into the center of the plate at a speed of 0.1-3.5 ml / min, while the spent eluent (eluate fractions) flowing to the bottom of the chamber pan due to centrifugal force is sent to the detector cell, the system is maintained for 1-3 min to establish a steady flow of eluent, apply a sample with a volume of 2.5 μl - 1.5 ml to the center of the plate or to a distance of 0-30 mm from the center of the plate, chromatography and continuous detection are carried out at a distance of 35-80 mm from the start.  2. The method according to p. 1, characterized in that the sample is applied to the plate before the rotation of the movable platform with the plate.  3. The method according to p. 1, characterized in that carry out gradient chromatography using eluent of variable composition. 4. A device for conducting thin-layer chromatography, consisting of a metal or polymer light-protective chromatographic chamber, which is a housing with a lid, with a chromatographic substrate plate made of glass, metal, polymer, and an adsorbent layer deposited on it, mounted on a support, and located inside the chamber body covered with a cover plate made of UV-transparent material, a device for feeding and removing eluent (eluate) from the plate, a light source with a light filter and a photodetector, excellent In that the chromatographic chamber has a cylindrical shape with a diameter of 290-310 mm and a height of 60-180 mm and is made in the form of a tray with a removable cover, there are holes in the tray: on the wall for inert gas, on the bottom in the center with a side for the mounting bracket pin, and the nozzle, the pallet has a groove at the bottom along the wall into which the eluate is collected and through the nozzle is pulled out or into the detector cuvette, and a recess in the wall for installing the cover plate, a multi-way dispensing tap is fixed in the center on the lid, connected with us the wasp, and 10-30 mm from it - the syringe dispenser, the nozzle of the metering valve and the needle of the syringe dispenser are inserted inside the chamber through special holes in the lid, a light source and a light filter, a photodetector or a light guide with a photodetector are built-in to it and installation above the chromatographic plate for detection at a distance of 35-80 mm from the start due to special holes or using a retractable frame of corrugated material; inside the chamber in the center at a distance of 5-50 mm from the base of the pallet, a horizontally movable metal platform with a diameter of 285-290 mm with a standard chromatographic plate of 100-200x100-200 mm in size mounted on it is fixed on a fastener pin connected to the motor a layer of adsorbent with a thickness of 100-5000 microns and a particle size of 3-30 microns, above the chromatographic plate is a cover plate with two holes for the spout of the dispensing tap and the needle of the syringe dispenser; the device is additionally equipped with a motor, the motor housing is tightly connected to the chromatographic chamber from below, the motor together with the camera is mounted on a support using a screw mount, which allows you to vary the position of the device at an angle of 0-45 ^o , outside the chromatographic chamber there is also a vessel with an eluent, a pump connected to a vessel with an eluent, a second photodetector, a cuvette and a second light source.