SU1404937A1 - Apparatus for thin film chromatography under pressure - Google Patents

Abstract

The invention makes it possible to expand the analytical capabilities of the device by introducing additional capillary fittings for introducing eluent located above the perimeter and the inner zone of the chromatographic plate. Each of the capillary fittings is connected through a flow regulator and flow booster to the eluent source, as well as through the flow regulator to the vacuum source. The eluent is fed to one or more capillary fittings. In order to provide the required trajectory for mixing, the samples through the other capillary fitting or several fittings select the eluate by connecting the fittings to the vacuum source. Thus, it is possible to achieve any trajectory of sample movement with an unlimited path of separation in any configuration of the eluent front. 8 il.

Description

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This invention relates to chromatography and can be used for the analysis of complex mixtures of vesicles.

The purpose of the invention is to expand the available capabilities of a device for thin-layer chromatography under pressure.

FIG. 1 shows a device, a longitudinal section; in fig. 2 - conditional scheme of the device; in fig. 3 shows one of the variants of the arrangement of capillary fittings above the surface of the chromatographic plate; in fig. 4-8 are chromatograms of separation of a complex mixture of organic substances.

A device for thin layer chromatography under pressure comprises a housing 1 with an internal flow cavity 2 for temperature control and pressure clamps 3.

A thin layer chromatographic plate 4 is placed on the housing 1 on which the membrane 5 is attached. On the housing 1 there is also a transparent cover 6 with a peripheral seal 7 so that a cavity 8 is formed between the inner surface of the cover 6 and the membrane 5 to create a back pressure on the membrane. Said cavity through a valve (not shown) communicates with a pressure source (not shown). Capillary fittings 9 for introducing the sample (one of them is shown) and capillary fittings 10.1, 10.2, ..., Y.n. for introducing eluent, located above the perimeter and the internal area of the chromatographic plate, are located in the lid 6, adjacent to the membrane 5 (FIG. 2 and 3).

Each of the capillary fittings 10.1, 10.2, ..., Yu.n. is connected (Fig. 2) through the flow controller 11 and the flow booster 12 to the source of 13 eluent, as well as through the flow controller 11 to the vacuum source 14. Capillary fittings 10.1, 10.2, ..., 10.n can be interconnected via flow regulators 11.

In the preferred embodiment, the flow controller 11, the flow boosters 12, and the vacuum source 14 are operatively connected to the control unit 15.

The device works as follows.

The sample of the mixture of substances to be separated is put on the chromatographic plate either preliminarily or through a capillary fitting 9.

In the cavity 8, a certain pressure of the medium (liquid or gas) is created by means of a valve. With the aid of the flow driver, 12, the eluent from the eluent source 13 is supplied under pressure (eluent pressure depends on pressure in cavity 8, sorption layer thickness and viscosity of eluent) to the capillary fitting 10.1 (or simultaneously to several capillary fittings). The composition of the eluent can vary according to any predetermined law, it can be

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various gradient compositions both with the help of a single driver 12 consumption, and with the help of the other boosters 12 consumption. In order to provide the required trajectory of sample movement through another capillary clamp (or several such nozzles at the same time), start the selection of the eluate, connecting them to the vacuum source 14. In this way, any sample trajectory and any configuration of the solvent front can be achieved; the length of the separation path may be unlimited.

Data on the order of connecting the fittings 10.1, ..., P. to the sources of eluent and dilution, on the composition of the eluent supplied and its required consumption is obtained as a result of preliminary calibration or detection results of the separation directly during the process, moving the transparent flap 6 end optical fiber optical detector. (In the case of short-circuited compounds, the movement of the eluted components can be observed directly, in the case of unstained compounds, with the help of marking dyes). In relatively simple cases, the separation adjustment can be done manually; in more complex operation of the device, block 15 controls the algorithm. In the course of separation, in order to obtain more compact chromatographic zones, it is possible to focus the zones with opposing eluent streams of the same or different composition, fed to the corresponding capillary fittings. With a sufficient number of capillary fittings, the chromatograph can carry out the separation according to all known variants of planar separation.

Example. The section of this mixture includes: a - 1,3 - dipalmitylglyceride; b - 1,2-dipalmityl glyceride; c - dipalmityl glyceride; d is cholesterol; e - palmitic acid; f is methyl palmitate; g - 2 monopalmitate; h - 1-monopalmitate; i-monogalactosyltriglyceride; j - cholesteryl palmitate; k is tetracosane; 1 - squalene; m - digalactosyldiglyceride; p - lecithin; about - lysolecithin. The separation is carried out on a layer of silica gel of size 20x20 cm. In FIG. 4-8, the location of the capillary fittings 10.1 -10.9 for the introduction of eluent is indicated by a cross, the eluted components of the mixture are indicated by letters.

At the first stage, fittings 10.1, 10.6 and 10.9 are connected to a source of eluent of chloroform - ethanol - trimethylborate (100: 1: 6), and fittings 10.3, 10.4 and 10.7 - to a vacuum source. After the separation in the first direction is completed (Fig. 4), the fittings 10.1, 10.2 and 10.3 are connected to the source of the eluent benzon-ethyl acetate-trimethyl borate (100: 20: 7,2), the fittings 10.7, 10.8 and 10.9 are connected to source of dilution, fittings 10.4 and 10.6 disconnect. After the process is completed in the second direction (Fig. 5), the fittings 10.2 and 10.8 are similarly disconnected, the fittings 10.1, 10.6 and 10.9 are connected to the vacuum source, and

10.3,10.4 and 10.7 - with a source of eluent of the composition heptane-benzene (3: 2). The result of separation in the third direction is shown in FIG. 6. Next shut off fittings 10.3,

10.4, 10.7 and 10.1, fitting 10.9 is connected to a source of eluent-heptane, and 10.6 is connected to a source of vacuum, which achieves separation of components k and 1 (Fig. 7). To separate the components t, p, and o, a chloroform-methanol-water eluent (69: 21: 2.6) is fed into the capillary nozzle 10.1, and the eluate is taken out through the nozzle 10.2 (the remaining nozzles are closed). The chromatogram of the separated mixture is shown in FIG. eight.

Claims (1)

Invention Formula A device for thin-layer chromatography under pressure, comprising a housing with an internal flow cavity for temperature control and pressure straps, a thin layer chromatography plate placed on the housing, a membrane adjacent to the chromatographic plate, a transparent cover with a peripheral seal mounted on the housing so that the inner surface of the lid and the membrane formed a cavity to create a back pressure on the membrane,  Capillary nipples for sample introduction and a capillary nipple for eluent input, located in the cap and adjacent to the membrane, connected to the eluent source through the flow regulator and flow rate stimulator, in order to expand the analytical capabilities, the device is equipped with additional capillary fittings for the introduction of eluent, connected to the sources of eluent through flow controllers and flow rate boosters and hch located above the perimeter and the inner zone of the chromatographic plate, all capillary fittings for introducing eluent are connected to the vacuum source through flow regulators. 0U2.2 Fig.Z if i i 7 / i // bjkx / fcf one Qf Fy Fie.6 Yu.7 FIG. five FIG. 7 FIG. eight