JPS5897655A - Automatic spotting apparatus for thin-layer chromatography - Google Patents

Abstract

PURPOSE:To obtain an automatic spotting apparatus capable of making a uniform analysis in a given time, by bringing a tip end part of a microsyringe needle into contact with a thin layer plate automatically. CONSTITUTION:A prescribed quantity of a material to be inspected is sucked by a microsyringe 1 set with a collar 13 at a constant distance from the tip end of a microsyringe needle 11 and said microsyringe 1 is inserted into a holder 3 and then, is fixed by a set bar 2. The needle 11 is pressed by a needle press bar 12 and is brought into contact with a thin layer plate 4. Next, dropping of the material to be inspected in the microsyringe 1 begins from the time when a press rod 9 contacts with a piston 8 by driving a driving part 6 and dropping down a movable shaft 7 and a press board 10. When the dropping is finished, a decent of the board 10 is stopped and the board 10 is returned to a starting position rising reversely and then, a spotting operation is completed. The uniform spot is obtained regardless of the experience of a worker and also, and the local heat decomposition of the body to be inspected is not caused by using this apparatus.

Description

[Detailed Description of the Invention] Thin-layer chromatographic densitometry is one of the methods to carry out the method, and when spotting a specimen on a thin hot plate, it is possible to spot-manipulate multiple points at the same time. Regarding the self-spot *alltK, which can be properly carried out under a constant sample speed 1 and a more uniform drying tube.

Conventionally, such spot operations have been mainly performed manually. In other words, hold the microsyringe that has aspirated the sample in one hand, hold the hair dryer in the other hand, and place the spot f on the thin plate, which has been determined in advance using a gauge, etc.
A very small amount of the sample in the litIC microsyringe,
! Complete the spot operation by applying drops and immediately drying with a dryer. Further, depending on the case, a large amount of specimen may be spotted by repeatedly dropping and drying the specimen at the same point. Generally 9 to 12 per layer board
When a point spot is formed and microquantity on the order of ng is required, K is several 10 for one spot.
The magnetic spot operation is returned *b three times by μl.

In this way, the spot operation is a simple and repetitive operation, but in order to maintain the precision of analysis, the operator must ensure that the diameter of the spot is uniform at a predetermined position on the thin plate. There is a need to perform spot operations like this.

Furthermore, the specimen spotted on the thin plate is subsequently expanded,
The substance in the sample is unstable and if left for a long time after the spot, the substance will physically or chemically decompose or denature, making accurate analysis impossible. , the time t required for spot operation must be shortened as much as possible.

In quantitative thin-layer chromatography, there is a limit to the number of thin-layer plates that can be processed, that is, the number of samples that can be processed.
Moreover, since there are restrictions on the opening during processing, a considerably high degree of maturity is required for accurate analysis.

An object of the present invention is to provide an automatic spot 5M value that can uniformly perform analysis within a certain period of time regardless of the skill level of the operator, and can greatly increase processing capacity.

The automatic spotting device for thin layer chromatography of the present invention, which has achieved this objective, is capable of attaching one or more microsyringes to the thin layer plate so that the sloped tip of the microsyringe needle and the thin layer plate surface are approximately parallel to each other. Microsyringe fixed flat membrane held at a predetermined inclination angle so as to face each other, microsyringe needle tip IIIA
An automatic specimen dropping machine W comprising a needle pressing means equipped with an elastic body that presses the microsyringe needle in order to bring the microsyringe needle into substantially even contact with the thin layer plate, and a piston pressing means that presses the piston of the microsyringe. It is assumed that tlL has the following.

In addition, the drying mechanism is also unique, and the thin layer plate is similar to Ki4.
A continuous air drying mechanism is provided, which has a mixing chamber that uniformly mixes the collected air to prevent overheating, and an air outlet that blows drying air to the thin layer plate.

Next, an embodiment of the automatic sub- jector of the present invention [Fig.
Although the detailed Km is used based on iK, the present invention is not limited to this example.

FIG. 1 is a partially cutaway schematic side view of an embodiment of the present invention.
The sample automatic dripping mechanism used by Hideki Ki is a microsyringe (
1), U micro syringe (1) T Sera) ) 4-(
2), a holder (8) to be held, a mounting base (5) for fixing the holder (3) at a predetermined angle of inclination, for example 15 degrees, to the thin plate (4), and a variable-speed front drive section. (6) movable shaft 7) a push plate (6) equipped with a push rod (9) installed at the free end of (7) to push the piston (8) of the microsyringe (1), and a pusher plate (6) for pushing the microsyringe needle Dlk; The tic basically consists of a needle pressing bar (2) whose tip is brought into contact with the laminated plate (4) with an appropriate force.

In the spot operation, move the microsyringe needle (B) so that the tip of the microsyringe needle (B) is held at a certain position 111.
6) Aspirate a predetermined amount of multiple samples from a microsyringe (1) with a collar (2) set at a certain distance from the tip of the microsyringe (1), insert it into the tube holder (?) and fix it with the setper (2). . After setting the holder (S) to which the microsyringe (1) is fixed onto the stand (6) K, press the microsyringe needle Qυ against the needle presser (2) K, and press the tip of the needle against the layer plate ( 4). The operations so far have been performed by hand, but these operations do not require skill.

Next, when the drive unit (6) is driven, the movable shaft (7)
The sample in the microsyringe (1) begins to drop when the pusher (9) comes into contact with the piston (8). The descending speed of the push plate (6) is set to a constant value based on the characteristics of the sample, such as viscosity. The push rod (9) pushes the piston (8) at a constant speed,
A predetermined amount of sample in the microsyringe (1) is transferred to a thin layer plate (4).
) When the dropping of K is completed, the push plate stops descending, then reverses upwards and returns to the starting position, completing the spotting operation. When using a plurality of microsyringes, there is a Δ variation in length, so if the lowering distance of the push plate is set as t - the total dripping distance of a certain microsyringe, 1IIIK, some microsyringes may not drip the entire amount. The descending distance of the rounded push plate (up to) must be determined to be the longest distance 111 at which the entire amount of the specimen in all microsyringes is dropped. However, in some microsyringes, the piston body may be pressed excessively and the microsyringe may be damaged. In order to prevent such damage to the microsyringe, the push rod (9) and the push plate are connected by a spring (b), and the piston (8) K is connected to prevent excessive pressure from being applied to the microsyringe. K11l has been constructed.

When the spotting operation is completed, the needle presser (2) is released and the laminate (4) is removed from the device and sent to the next unfolding operation. In addition, in the case of a spot operation that is repeatedly performed in batches of several tens of microliters in the determination of wrinkles in the NY order, when the first spot H1 is completed, the needle pressing bar (6) is released and the holder (3) held in the spot is released. Set lever (
2) and remove each microsyringe (1), aspirate the prepared specimen, and then insert the microsyringe (1).
1)? Insert it into the holder (3) again, fix it to the base (I), and repeat the slit operation described above.

In the spot operation, the degree of contact between the tip of the microsyringe needle (b) and the layered plate (4) is an important factor for producing a spot with positive aIK and uniform size. This is because when dropping a minute amount of sample, such as a few microliters, the tip of the microsyringe needle is
A K droplet is formed. The size of the droplet varies depending on the state in which the microsyringe is held, that is, whether it is upright, tilted, or lying on its side. We must grow until we can overcome this. The spots seen by such natural dripping are not uniform and are very large, making it difficult to develop, select, and analyze the sample in many cases. In order to avoid this drawback, if you do it manually, use the microsyringe holding shape gsc111I.
* Without damaging the thin layer, the tip of the microsyringe needle is brought into contact with or close to the thin layer at a 4-degree angle to allow the droplet to land on the thin layer. However, this operation requires considerable skill, and even if sufficient care is taken, it is impossible to always obtain a uniform spot.

In the present invention, as shown in FIG. 2, such a complicated operation is carried out on the microsyringe (1) so that the sloped tip of the microsyringe needle (b) and the thin plate (4) If face each other in parallel. T-Installation is done by holding the stand (5) K and further pressing the microsyringe needle (b) with the needle presser (6) so that the tip of the needle aυ comes into light contact with the thin plate.
It is designed to be easy to do. An elastic body (1) made of synthetic rubber or the like is attached to the needle presser (2) at the part that contacts the microsyringe needle (2).
The elasticity of the elastic body (2) changes the pressing force to an extent that does not damage the surface of the thin laminate. At this time, a certain gap (d) is formed between the slope of the tip of the needle (b) and the surface of the thin layer.
The sample discharged at a constant speed is deposited on the thin plate before a single drop is formed into a tube, and the spot size becomes small and uniform.

Even when forming spots at multiple points in the housing, the elastic body (2) of the needle suppression bar (2) absorbs the variation in the degree of distortion caused by the non-uniform length of the microsyringe needle aυ. Equal contact between the tip of the needle (b) and the laminar plate is obtained, so that every spot is uniform.

In this embodiment, an example is given in which one microsyringe is fixed, but two or more microsyringes can be attached and fixed to the stand at the same time to perform a plurality of spot operations at the same time.

Another important factor in obtaining a uniform plate is the drying of the sample dropped onto the thin plate. Conventionally, as mentioned above, the sample that has been dropped is immediately dried using hot air from a dryer or a heat plate, resulting in a larger spot. However, when such a method is used, unless consideration is given to the heat distribution of the generated heat source, there is a risk of locally high heat generation and, depending on the type of specimen, decomposition and p-denaturation.

In the spotting device of the present invention, as shown in FIG. 1, the mixing wIQη and the mixing ii! are provided with a diffuser plate inside. (b) Drying air with uniform temperature distribution inside the tVW plate (4) The nine continuous air dryer board is basically composed of the air outlet (2) that blows the air, and the plate is placed on the back side of the thin layer plate (4). By placing the KLA in close proximity to each other, the drying rate of each dropped specimen can be adjusted to the same level to form a uniform spot.

Mixed ii! (B) is formed within the installation base (to) of the thin layer plate (4), and an opening is provided in the upper surface of the installation base (2). Therefore, by placing the thin plate (4) on the installation stand (6), the opening is closed, and drying air is sent out only from the air outlet.

The drying air is supplied from the blower 4. The air is supplied from the air blower ms (e). The air is heated to a predetermined temperature by the heater (b), and after being purified by the dust removal filter 4, it is passed through the conduit @. Mixed fit 07) K is sent in. In the mixing w1v), the sent drying air collides with the diffusion plate 94, diffuses radially, mixes with the air in the mixing chamber αη, and then is sent out from the entire surface of the air outlet. Ventilation port (to)
is open near the base of the microsyringe needle Qυ, that is, near the microsyringe (1) where the dropped specimen is also exposed. In addition, the air outlet (to) is long in the direction perpendicular to the direction of the flame force of the thin layer plate (4), and the opening area is smaller than the cross-sectional area of the mixed wlaη, so it can be used as a drying air outlet. With positive pressure, a uniform air volume can be obtained over the entire surface of the air outlet.

In this way, the drying air blown from the air outlet (to) has a uniform temperature distribution and the same air volume in all parts, so it is possible to achieve a local high #I! II does not occur, and the nine samples of solvent dropped onto the thin plate evaporate uniformly, and all spots are dried at a constant rate, so the spots seen are of equal size.

In the spotting apparatus of the present invention, the spot size t can be freely controlled by appropriately selecting the dropping rate of the specimen, the amount of drying metal, and the temperature.

As described above, when using the spotting device of the present invention,
A uniform spot can be obtained regardless of the operator's experience and without local thermal decomposition of the specimen.

[Brief explanation of the drawing]

Fig. 1 is a -S notch schematic side view of a complete example of the automatic spotting device of the present invention, and Fig. 2 is a schematic explanatory diagram showing the state in which the microsyringe needle is pressed by needle pressing (-). Yes. (Main symbols in the drawing) (1): Micro syringe (3): Holder (4): Thin plate (b): Mounting base (9): Push keyaki αQ: Push plate Qv = Micro syringe needle 42 Needle suppression par o3: Elastic body (lL7): Mixing chamber (to): Ventilation port

Claims (1)

[Scope of Claims] Eleven or two or more microsyringes are held at a predetermined inclination angle to a thin laminate so that the sloped tip of the microsyringe needle and the thin laminate face are substantially parallel to each other. Microsyringe fixing means, microsyringe needle tip at thin layer plate! A thin layer comprising an automatic specimen dropping mechanism consisting of a needle pressing means equipped with an elastic body that presses the microsyringe needle to bring the microsyringe needle into qualitatively uniform contact, and a piston suppressing means that presses the piston of the microsyringe. Automatic spotting device for chromatography. 2. Special feature where the inclination angle of the microsyringe is about 15°! ff--A device according to claim 1. A microsyringe fixing means for holding 31 or more microsyringes at a predetermined inclination angle so that the processed tip MWi of the microsyringe needle and the thin layer plate surface are opposed to each other in parallel to each other. An elastic body is provided to suppress the microsyringe needle in order to bring it into substantially uniform contact with the WAt thin plate at the top end of the syringe, and a sample automation system consisting of a suitable suppression means and a piston suppression mechanism that presses the piston of the microsyringe. An automatic spotting device for thin layer chromatography comprising a dripping mechanism, a mixing chamber for uniformly mixing drying air, and a continuous air blow drying mechanism provided at an air outlet 1 for blowing collected air to a layered plate. 4. The device according to item 3, wherein the microsyringe has an oblique angle of about 15°. 5. The installation according to claim 3, wherein the continuous air drying mechanism is provided near the back surface of the thin laminate. 6. Claim 5, wherein the air outlet of the continuous air drying mechanism is provided near the mounting base of the microsyringe needle.
Apparatus described in section.