CS230581B2 - Plate for shallow layer pressure chromatography - Google Patents

Description

The disadvantage of classical thin layer chromatography is the long development time due to the short layered plate, the division is not perfect and the theoretical number of plates is limited.

Significant progress has been made in the development of high performance thin layer chromatography (HPTLC) (A. Zkatkis and R. E. Keiser: High Performance Thin-Layer Chromatography, Elsevier, 1977). Compared to conventional thin-layer chromatography, a better separation is achieved at a shorter elution time and not a shorter distance.

The latest technique, however, is the so-called pressure thin-layer chromatography (Hungarian patent! 173 749), which can be carried out on both thin and thick layers and the separation is the fastest. In TLC, the solvent system flows through an adsorbent layer in a closed system due to overpressure. The entire adsorbent layer is preferably covered with an elastic film which is pressed against the adsorbent layer by external pressure. The pressure of 0.2 to 1.0 MPa with which the film is pressed is always higher than the pressure of 0.1 to 0.8 MPa below which the liquid component flows. In comparison with the previously known planar chromatographic methods, a better separation in the so-called pressure ultremicron chamber is achieved

2305-1 for a very short time of 1 to 10 minutes. The apparatus was designed for both concentric and linear chromatograms.

The basic problem of the linear alternative of thin-layer chromatography is that the solvent component escapes at the edges of the chromatographic return and that the solvent queue is not straightforward because there are no concentric, semi-circular or quarter-circular queue lines at the solvent, the solvent is fed at one point.

It is an object of the present invention to provide a thin-layer chromatography plate which can be used in a pressure-thin-layer chromatography apparatus, in that the developing solvent component does not leak at the edges of the plate and 8 by passing the solvent component through a straight sele queue.

The invention is based on the finding that when the edsorbent layer is sealed in an appropriate manner at the edges of the layered board, the solvent does not escape during the slack.

The essence of a thin-layer pressure chromatography plate having a layer of Bdsorbent deposited thereon is that the edsorbent layer is sealed on at least two edges of the plate and, at or where the solvent component is introduced in at least one zone, the edsorbent layer is removed from the carrier plate. .

The adsorbent layer may be removed in one or more thin zones. It is also possible to place a thin baffle plate, plates or wires directly into the solvent feed line of the solvent feed line to form a solvent chamber with the solvent component, thereby ensuring that the front of the solvent component queue is straight.

The adsorbent layer of the present invention is sealed by impregnation, or the adsorbent layer is compressed using powdered glass, where the compression is different at the edges of the layer and in the chromo-gravel field. Alternatively, one or more parallel grooves are formed in the carrier plate, the same carrier plate being sealed at the edges or, in the case of a ceramic carrier plate, provided with glaze ends thereof, for example tin oxide white.

Materials for impregnation or glaze should be insoluble in the solvent used in chromstography, should not be changed due to the excess pressure of the solvent component and given temperature conditions, and there should be no chemical reactions between them with the adsorbent layer, foils, or sealing the adsorbent layer .

As impregnating material, paraffin, synthetic solutions or suspensions and water glass, i.e. sodium silicate, are preferably used. The edges of the ceramic plates ae may cover the glaze of tin-lead white. If powdered glass is used for compression, then glass of different quality is used at the edges of the carrier plate, or other edltive may be used to seal the edges of the carrier plate.

In one-dimensional development, it is sufficient if the two edges of the layer are sealed and the side opposite to the direction of advance of the queue. However, in two-dimensional development, it is necessary to seal the layer on all four sides. In a two-dimensional linear development using an intermediate solvent, it is sufficient to seal the two opposite edges of the board layer, preferably the two longer sides.

the width of the sealing layer may be between 0.05 and 3 cm. Most preferably, the layer is 1 cm or thinner. The length of the layer is the same as the distance of the chromatographic layer.

The straight front of the developing solvent component queue is achieved in accordance with the invention by removing an adsorbent layer 0.1 to 0.5 mm wide in one or more zones at or near the solvent feeding point. The maximum length of the grooves thus formed is the same as the length of the adsorbent layer which remains after covering the ends of the plate, but is preferably 1 to 20 mm shorter on both sides of the support plate. The grooves may be arranged side by side. The maximum depth of the grooves is not greater than the thickness of the adsorbent layer. The number of grooves may preferably be between 1 and 4.

Wire or wires with a diameter of 0.05 to 1.0 mm or baffle plates may be placed in front of the sample application site or sites. The maximum length of the baffle plates or wires after covering the edges of the plates is the same as the length of the adsorbent layer, but is preferably shorter by 1 to 20 mm on both sides of the plate. The baffle plates or wires may be placed side by side. Their number is preferably from 1 to 4. Preferably, a gapless synthetic plate fixed by glazing the plate layer is used.

A so-called concentration zone can also be formed in the part of the solvent supply to the plate layer which has been produced as mentioned above. That is, an additional layer zone is applied to the adsorbent layer. The material used for this layer is, for example, silica earth, powdered glass, synthetic powder or cellite, i.e. they are carriers rather than adsorbents used for separation, such as alumina or silica gel. However, in low porous fine grain layers, high porous silica gel with a wide pore size range is occasionally used. The double layer thus produced facilitates the concentration of the sample.

For the chromatographic plate layer according to the invention, preferably a fine grain layer having a grain size of 0.5 to 10 µm or a normal grain size of 10 to 40 µm is used. Any commercially available layered board may be treated according to the invention for thin layer pressure chromatography. However, the adsorbent layer can also be produced non-commercially, directly in the laboratory.

The novel b higher effect of the plates produced according to the invention lies in the fact that in the case of overpressure, the developing solvent component does not leak at the edges of the plate and longer in that the solvent component proceeds with a straight front of the queue. Until now, this effect has not been achieved with the known thin layer chromatography plates.

An exemplary embodiment of the invention is shown in the accompanying drawings, in which: FIG. 1 is a schematic diagram of a TLC plate with a silica gel layer; FIG. 2 is a schematic diagram of a TLC plate with an edge-impregnated silica gel layer; Fig. 3 shows a thin-layer pressure chromatography plate with an impregnated edge and a concentration zone; Fig. 4 shows a thin-layer pressure chromatography plate with a magnesium silicate layer impregnated on four sides, in which the solvent is regulated in both dimensions by a synthetic film; Fig. 5 shows a thin-layer pressure chromatography plate in which the adsorbent layer is embedded in grooves formed in the support plate.

Fig. 1 shows a thin-layer pressure chromatography plate provided with a solid paraffin-impregnated silica gel layer (1). At the front of the solvent inlet g there is a steel wire behind which the sample application points 2 are arranged.

FIG. 2 shows a thin-layer pressure chromatography plate having grooves arranged to control the progress of the solvent in the layer.

The thin-layer pressure chromatography plate shown in FIG. 3 has impregnated edges 6 and a concentration zone 11.

23058,

Giant. 4 shows a thin-layer pressure chromatography plate in a two-dimensional development arrangement with a magnesium silicate absorbent layer (8), where after one-dimensional development in the direction of arrow e, the plate is impregnated on the fourth side and the development proceeds in the direction of arrow. The solvent supply and progression are controlled by synthetic film 2 ·

Not shown in Fig. 5 is a thin-layer printing chromatography plate in a five-solution arrangement, with barriers 11 arranged at the edge of the edeorbent layer 10. formed from a carrier plate. The barriers 11 separate the individual grooves 2 from each other.

The invention is further illustrated by the following examples:

He did

1.0 ml of a 0.85% aqueous solution of a high-polymer polyacrylamide homopolymer is added to 32 g of silica gel powder having a grain size of 4 to 8 .mu.m and an inner diameter of 600 nm. After mixing, the properly cleaned plates, 20 x 20 cm glass panels and a thickness of 1.5 m are applied in a known manner in a thickness of 0.25 mm. The layers were dried at room temperature. The produced layers have good mechanical resistance even after activation.

The ends of the layered sheets thus produced and activated are impregnated on three sides in width, 0 and immersed in paraffin, which melts at 130 ° C.

To guide the solvent process, a 0.2 mm diameter steel wire 6 is placed on the Selo solvent inlet 2 as shown in Fig. 1. Authentic coloring, for example, butter gel, Sudan G, indophenol, can be well placed in an ultramicron chamber at pressure of 1.0 MPa, elution time 10 minutes, elution path length 15 cm, elution was carried out with methylene chloride.

Example 2

The procedure is as in Example 1 except that 30 g of neutral alumina is used instead of 32 g of silica gel powder. The grain size is 15 to 20 µm, the thickness of the deposited layer is 0.3 mm.

Example 3

100 ml of a 1.3% aqueous solution of an anionic acrylamide-acrylic acid polymer mixture was added to a 4 to 5 Aid silica gel powder having an inner grain diameter of 600 nm. After mixing, the suspension is applied in a thickness of 0.3 mm to a 20 x 40 cm polyterephthalic film and a thickness of 0.4 mm. The layers dried at 75 ° C have adequate resistance even after activation. Immediately after the application of the layer and through the drying described above, 2 mm wide, 0.3 mm thick and 85 mm are placed at the front of the solvent inlet 2, 15 mm from the odd impregnated board edge and equidistant from the other two edges to be impregnated a long polyterephthalic film to control the flow of the solvent (see schematic figure 1). This plate is adhered to the layer during drying.

The edges of the three sides of the layered sheets thus dried are impregnated with the control board or the foil by soaking in a 1.5% aqueous solution of a high-polymer, non-ionic polyacrylamide homopolymer in a width of 5 mm. It is then dried at room temperature.

On the layer thus produced, the digital glycosides were effectively separated within 18.5 minutes in a linear pressure ultramicron chamber, eluting with 2-butanone over a distance of 35 cm, a pressure of 1.0 MPa.

Example

The procedure of Example 2 is followed except that two grooves 2t are formed in the layer with respect to solvent control, each of which is 3 mm wide (see schematic figure 2).

Example 5

100 ml of a 2% aqueous solution of polyvinyl acetate was added to 26 g of a silica gel powder having a grain size of 8-10 µm and an inner diameter of 600 nm. At the same time, 30 ml of a 2% aqueous polyvinyl acetate suspension was added to 8 g of silica of a grain size of 10-20 .mu.m.

The two systems are homogenized separately in an electric homogenizer for one minute. The two suspensions are then poured into two troughs, suitably spaced in terms of space and length, and applied in the usual manner to aluminum foils of 20 x 20 cm and thickness of 0.2 mm. In this way, a 0.25 mm thick layer with two adsorbent zones is produced. The width of the inactive zone, depending on the groove setting, is 40 nm (Fig. 3).

Before drying of the adsorbent layer, immediately after application, 15 mm from the odd impregnable edge of the plate, facing away from the side of the inactivated adsorbent zone, and at the same distance from the other two parallel edges to be solvent, impregnate, place a 0.3 mm thick, 1 mm wide and 160 mm long polyterephthalene plate in order to control the solvent flow thereafter. The layer is then dried and optionally activated. In this case, the board edge impregnation is carried out as described in Example 2 above.

On the so-called concentration zone 2 layer, samples of volatile oils obtained from inflorescence of Matricaria chamomilla L. can be efficiently distributed over 12 minutes by steaming. The process is carried out in a linear pressure chamber, eluting with benzene, a pressure of 0.7 MPa, a solvent velocity of 20 ml / h, eluting to a distance of 15 cm.

Example

The procedure is identical to the procedure described in Example 4 except that instead of 8 g of silica clay, 7 g of cellulose powder having a grain size of 10 to 20 µm is used.

Example 7 g of powdered magnesium silicate having a grain size of 6 to 10 µm together with 110 g of an aqueous solution of sodium silicate having a density of 1.042 g / cn? measured at 15 ° C homogenizes the crushers in trams. Thereafter, a suitably cleaned 1.5 mm thick glass panel is applied in a conventional manner. The plates were stored for 2.5 hours in a 75% humidity chamber. Then they are removed at room temperature and dried. In this way a stable layer is produced.

The edges of the sheets thus produced are impregnated as described in Example 1 above. These sheets can be used for efficient dye distribution.

Example e

The procedure is as in Example 3, except that the two opposite edges of the layer are impregnated by immersion in a 1.5% aqueous solution of a high polymeric nonionic polyacrylamide homopolymer prior to drying the adsorbent layer 10. At the same time, a 1 mm thick groove 2 is produced in the center of the adsorbent layer 10 at the solvent supply point, followed by solvent flow control and two-dimensional developments. Then the adsorbent layer 10 is dried and used for chromatography.

Example 9

The procedure of Example 7 is followed except that the adsorbent layer is adapted for two-dimensional development. After one-dimensional development, the board is impregnated with a fourth wiped coating. The solvent is regulated as shown in Figure 4.

He did

Six grooves χ deep 0.3 mm are formed in an aluminum plate 20 x 20 cm, 2 mm thick. The grooves 1 are filled with a paste made of 8 to 1 µm powdered polyamide 11 (25 g of polyamide powder are mixed in 100 ml of a 1.5% aqueous solution of polyvinyl acetate and the solution is then homogenized for 1 minute.

A 15 mm wide barrier 11 is produced at the edges of the adsorbent layer 10. The barriers 11 which separate the individual grooves χ from each other are made by smoothing the surface at a width of 10 mm. A synthetic polyethylene film 2 of 16 x 3 x 0.3 mm, as shown in Figure 5, is placed in the Sela of the five solvent delivery points χ, as shown in Figure 5. This ensures a straight line of solvent. Samples are applied 2 mm from the solvent inlet X opening.

Example

The procedure was as in Example 1 to 5, wherein the silica gel used contained manganese-activated zinc silicate as a fluorescent material.

Claims (8)

SUBJECT OF THE INVENTION A thin-layer chromatographic plate having an adsorbent layer deposited on a plate, characterized in that the adsorbent layer (10) is sealed at least on two edges of the plate and at least one zone at least one zone at the solvent inlet (3). 10) adsorbent is removed from the plate, A plate according to claim 1, characterized in that a partition, preferably in the form of a plate or wire, for example a steel wire (4), is provided at the point of solvent supply (3) to form the solvent chamber. The board according to claim 1, characterized in that the sealing of the adsorbent layer (10) is effected by impregnation, for example with paraffin or sodium silicate. 4. The board according to claim 1, characterized in that the sealing of the adsorbent layer (10) is carried out by compressing it with powdered glass. The plate according to claim 1, characterized in that a groove (5) is formed in the adsorbent layer (10) and the adsorbent layer (10) is sealed at the edges of the plate. 6. The board according to claim 1, characterized in that at least two parallel grooves (5) are formed in the adsorbent layer (10) for the two-fold development, and the adsorbent layer (10) is sealed at the edges of the board. A board according to claim 1, characterized in that it is ceramic and its edges are provided with a glaze, for example white lead, for sealing the adsorbent layer (10). A plate according to claim 1, characterized in that it is provided with an inactive concentration zone (7), for example of silica clay, powdered glass or cellite.