SE454729B - Closed system thin layer chromatography appts. - Google Patents

Abstract

The thin layer chromatography plate (25) is contained within a sealed system. under a cover (32). The plate is conditional by a moist gas which passes through pipes (33) (35) (36). When the plate has achieved the desired moisture content, the is supplied to the plate via chamber (28).

Description

10 15 20 25 30 454 729 with variations in air humidity, which among other things results obtained are not exactly reproducible. only.

An object of the present invention is now to provide provide a means of chromatography, which provides repro duceable results.

Another object of the present invention is to provide a chromatography device that can be used for a number of different types of common chromato- which make it possible to achieve precise reproduction duceable results, regardless of the specific technology used.

These and other objects of the present invention are achieved with the method of the present invention, which by placing a thin-layer plate with applied samples race in a chamber, attenuated tightly closed to form a closed system, that the plate is conditioned to the desired moisture by means of a gas which is led into and out of the chamber. maren, and that after reaching equilibrium eluent is added and brought into contact with the plate.

In this way, the humidity of the plate can be regulated via gas phase and thereby the silicon dioxide layer’s own creates are maintained constant, even when chromatographed takes place on different occasions.

According to an embodiment of the invention, the layer plate with the active layer downwards over a space me in the chamber, after which the chamber is tightly closed to of a closed system, the plate is then conditioned to the desired humidity by means of a gas which is led into and out of the space under the plate, after which achieved equilibrium eluent is applied and brought into contact with the plate 10 15 20 25 30 454 729 edge. After conditioning to the desired humidity, the saturation is achieved by the addition of a solution means to the space under the thin-layer plate. The big the advantage of this is that the saturation time can be varied before the chromatography is started.

According to another embodiment of the invention is placed the thin layer plate in the chamber with the active layer upwards, after which the chamber is tightly closed to form a locking tet system. The conditioning is then done by gas guided through the chamber through the gap between the plate and one chamber closing lid until desired humidity is reached.

This makes it possible to introduce gas, for example into in the form of solvents or ammonia, after conditioning ring, which provides increased separation possibilities. By as a lid use a coverslip can chromatography of colored substances directly observed. About the cover plate replaced with a quartz glass cover plate can be UV-absorbent substances are studied with UV lamp. For UV-sensitive substances see, the invention also provides the opportunity to work under to nitrogen atmosphere, whereby the substances can be das.

According to yet another embodiment of the invention the thin layer plate is dried after chromatography according to any of the foregoing embodiments, in order to placed in a closed chamber with the layer down, each on the eluent is brought into contact with a selected area of the plate to specifically chromatograph the selected part by placing an absorbent layer material in the space under the plate in the eluent and in contact with a_nt- selected area of the plate, perpendicular to the elution direction ningen.

The device according to the present invention precedes the island 10 15 20 25 454 729 thin layer chromatography to obtain reproducible results are mainly characterized by a tight bar chamber, a space arranged in the chamber intended to covered by a thin layer plate, into said space opening the gas and liquid supply and discharge means, at least one receiving space arranged in the chamber for elution liquid with means for bringing the eluent into con- pace with the thin layer plate, means of pre-supply and release of liquid in said receiving space and means for gas supply and gas emissions in the chamber. Benefits of the device appears from the above investigation in connection to the method according to the invention.

According to a suitable embodiment of the device are arranged two uptake compartments for eluent on two counter- standing sides of the space over which the thin-layer plate tan is intended to be placed. This allows chromatography take place from two directions on one and the same plate, which leads to that the time required for chromatography of a series of samples can be significantly shortened.

The advantages of the invention may be summarized as follows: jande: A large number of techniques known per se can be performed with the same instrument.

Complete control of humidity and humidity sea saturation.

Oxidation-sensitive substances can be chromatographed in gas atmosphere. .

The chromatographic process can be studied directly. 10 15 20 25 30 454 729 With the device according to the invention, the separation can further by gas phase variation.

Finally, the device according to the invention is very light. worked and is well suited for research and teaching purposes and for routine analyzes.

Additional purposes, benefits and characteristics of the present invention will become apparent from the following detailed description in connection with the accompanying drawings, on which Fig. 1A shows a plan view of the device according to the invention, Fig. 1B shows a perspective view of the device according to the finning with a thin-layer plate laid in, and Figs. 2-5 show the operating principle in the implementation of layer chromatography according to different techniques.

The device according to the invention is denoted by 1 in Fig. 1 and comprises an outer edge area 2, a chamfer 3, against which a lid 51, for example in the form of a coverslip disc sealing bearing. A trough-shaped space 4 is raised taken in the central part of the device, with two opposite walls 5 and 6, the upper edges 7, 8 of which are located below the level of the outer edge area of the device.

In chromatography, the thin layer plate is placed on the said upper edges 7, 8 and then end up with a suitable design of the device at a distance of about 1 mm from the lid underside.

Outside the central boundary of the opposite area, 5, 6 are arranged receiving spaces 9, 10 for eluent with means for bringing the eluent into said receiving means. the space in contact with the thin layer chromatography plate in shape 10 15 20 25 30 454 729 a glass plate 11, 12 which by means of a screw 13, 14 can moved to contact with the thin layer plate. An advantage is that only a very small amount of eluent is required race.

In the central space, connections 15, 16 open for access. supply and discharge of gas / liquid. ' In each of the receiving spaces for eluent mouths a liquid supply connection 17, 18. Furthermore, means for supply and emission of gas arranged in connection to said spaces in the form of a conduit 19, 20 with a plurality evenly distributed across the width of the device. supply channels 21, 22. In order to achieve a lower pressure case over the device, the channels used for gas emissions are designed with larger diameters.

In the embodiment of the device according to Fig. 1 shown the invention is furthermore arranged as suitable means 23, 24 which press the lid to a sealing abutment towards the outer edge area 2 of the device, but of course can other solutions are also considered in this regard.

Working principle of the chromatography apparatus according to the invention is shown in more detail in Fig. 2, which shows a cross-sectional view through the device. A thin layer plate 25 is laid on two walls 26, 27 over a central trough format space 28. Platumx chromatographed in the shown execution only from one side. (the one on the right in the figure), wherein the glass plate 29 in the eluent space 30 by means of the screw 31 bring into contact with the edge of the plate. Of course can__ a thin layer plate is chromatographed from both edges at the same time. The system works with a closed chamber, by a coverslip sheet 32 or equivalent sealing has been applied towards the upper edge area of the device. Gas supply lines 5 10 15 20 25 30 454 729 to the respective eluent space has been indicated by the ducts 33, 34 and the air supply ducts 35, 36.

When the chromatography is to be started, the plate 25 is placed after sample application with the layer downwards over the central space 28. The plate is then conditioned to the desired increased humidity by supplying gas to the respective taken out of the central space via connections indicated at 37 and 38, respectively. When equilibrium is obtained, it is added eluent 39 in the trough 30 and in this case only a tal milliliter. The chromatography is started by the disc 29 with the screw 31 is brought into contact with the thin layer plate 25. If chamber saturation is desired, fill in the space 28 a few milliliters of the desired solvent. The saturation time can then be varied before the chromatography is started, which can be beneficial. In addition, there is opportunity to vary the composition of this liquid. The process is interrupted, either by the end of the eluent or by the by pushing away the cover plate and the chromatography plate lifted out. This technology is also called N-mode.

According to an alternative approach, the the layer plate 25 with the active layer upwards and conditioned the ionization takes place by gas supply through, for example line 33 and ducts 35 while the gas is being removed through channels 36 and line 34. Chromatography the ring is started and interrupted as above. Space 28 is in this case unused, but can for example be used for thermostating the thin layer plate. An opportunity that opens with this construction is to introduce gas, to for example ammonia or solvent vapor, depending on the tionation, which provides increased separation possibilities. Far- With this technology also known as S-mode, crematorium graphing of colored substances is directly observed. About one quartz glass board is used as a cover plate, UV 10 15 20 25 454 729 * -absorbent substances are studied under UV lamp. UV- light-sensitive substances can be protected by: maintain a nitrogen atmosphere in the chamber.

Figures 3 and 4 show the principle of continuous production. cooling in S-mode (Fig. 3) and N-mode (Fig. 4). In S-mode a shorter cover plate 40 is used, whereby the eluent can is made to evaporate to the desired distance by firing the cover plate to a suitable position. Here, too, gas can be passed through the conduit 41 and the channels 42 to achieve desired chromatographic selectivity.

In N mode, the position of the thin layer plate 43 can be varied and maren can still be tightly closed by means of the cover plate 44. In this case, eluent 45 can be filled in the central space 46. In this case, it is advantageous to use one quartz glass plate to study the process under UV light.

Fig. 5 shows the principle of "Mobile Rf" mode, one hitherto relatively unusual form of chromatography, which is used for to specifically chromatograph a selected portion of a thin layer plate, which has been dried after chromatography according to any of the methods presented above.

This brings eluent 47, which has been introduced into the central space 48, in contact with the selected part of the thin layer plate 49 via, for example, a folded filter paper 50 or other absorbent material as placed at an angle the right motelution direction. This can suitably be fixed by means of Teflon rods. The process is started by supply of eluent to the space via the previously mentioned the fluid supply connection.

A6

Claims (15)

10 15 20 25 454 729 P a t e n t k r a v A method of producing reproducible thin layer chromatography results, characterized in that a thin layer plate with applied samples is placed in a chamber, that the chamber is tightly closed to form a closed system, that the plate is conditioned to the desired humidity by means of a gas which led into and out of the chamber, and that after reaching equilibrium eluent is supplied and brought into contact with the plate. 2. A method according to claim 1, characterized in that the thin layer plate is placed with the active layer downwards over a space in the chamber, after which the chamber is tightly closed to form a closed system, that the plate is conditioned to the desired moisture by means of a gas which is led in and out of the space under the plate and that after reaching equilibrium eluent is added and brought into contact with the edge of the plate. 3. A method according to claim 2, characterized in that after conditioning to the desired humidity, chamber saturation is achieved by adding a solvent to the space under the thin layer plate and that chromatography is started only after the desired saturation time has elapsed. 4. A method according to claim 1, characterized in that the thin-layer plate is placed in the chamber with the active layer upwards, after which the chamber is tightly closed to form a closed system, that conditioning takes place by. gas is passed through the chamber through a gap between the plate and a lid closing the chamber to the desired humidity and that after reaching equilibrium eluent is supplied and brought into contact with the edge of the plate. 10 15 20 25 30 454 729 10 S. A method according to claim 4, characterized in that a gas such as ammonia, solvent vapor, is introduced into the chamber after conditioning to increase the separation possibilities. 6. A method according to claims 4 - 5, characterized in that the chamber is closed by means of a cover glass sheet. 7. A method according to claims 4 - 5, characterized in that the foam of quartz glass is used to tightly seal the chamber, wherein nitrogen gas is used as a shielding gas atmosphere to protect UV light-sensitive substances when the process is followed under UV light. _ _ 8. A method according to claims 1 - 7, characterized in that the thin layer plate after chromatography is dried and then placed in a closed chamber with the layer downwards, and that the eluent is brought into contact with a selected area of the plate to specifically chromatograph only a selected part of this by placing an absorbent sheet material in the space below the plate in eluent and in contact with the selected area of the plate, perpendicular to the direction of elution. Device for performing thin layer chromatography to obtain reproducible results, characterized by a tightly closable chamber (1), a space (4; 28; 46; 48) arranged in the chamber, intended to be covered by a thin layer plate (25). : 43; 49), into the said space opening supply and discharge means (l5, _l6; 37, 38) for gas and liquid, at least one receiving space (9, 10; 30) for elution liquid with means arranged in the chamber (13, 14; 31) for bringing the eluent into contact with the thin-layer plate, means (17, 18) for supplying and discharging liquid in said receiving space and means h! IS 20 25 454 729 ll (19, 20, 21, 22; 33, 34, 35, 36; 41, 42) for gas supply and gas emissions in the chamber. Device according to claim 9, characterized in that the chamber has a detachable part, tightly closing lid (32; 40; 44) in the form of a cover glass sheet. ll. Device according to claim 9, characterized in that the lid (32; 40; 44) is arranged to be at a certain distance above and parallel to the thin-layer plate (25; 43; 49). ' 12. ' Device according to claim ll, characterized in that said distance amounts to approximately 1 mm. Device according to claims 10 - 12, characterized in that the lid (32: 40; 44) consists of a quartz glass plate. Device according to claim 9, characterized in that the receiving spaces (9, 10; 30) for elution liquid are arranged on two opposite sides of the space (4; 28; 46; 48) over which the thin-layer plate (25; 43; 49) is intended to be placed. Device according to claim 9, characterized in that the means for bringing the eluent into contact with the thin-layer plate (25: 43; 49) consists of a movably arranged disc (11, 12; 29), which by means of screw means (13, 14; 31) or the equivalent is pressed against a wall (5, 6; 26, 27) adjacent to the space on the edge (7, 8) of which the thin-layer plate rests.