JPS6095357A - Analyzing device for crinical analysis - Google Patents

Abstract

PURPOSE:To eliminate the need for standing, serum sepn., dispensing, etc. in a device for analyzing clinically a blood, urine, etc. by enabling analysis with a single device right after blood gathering. CONSTITUTION:The chamber in a base body 5 formed of glass, etc. is partitioned by a dialysis membrane 9 to the 1st chamber 6 and the 2nd chamber 7. A sample liquid such as blood or the like is introduced through an introducing port 10 into the 1st chamber 6 and sample molecules (sugar) are passed through the membrane 9 and are conducted into the 2nd chamber. An immobilized enzyme 12 is packed in the 2nd chamber and an enzyme reaction with the sample molecules is effected. A buffer soln. is then introduced through an introducing port 13 to color the sample subjected to the enzyme reaction. The colored sample liquid is introduced into a detecting cell part 8 and the colorimetry is accomplished by a photodetector 19 such as a photodiode, etc. Since the process of blood cell sepn.- reaction-measurement is accomplished with the single device, direct analysis is made possible. The need for operations such as standing of the blood, serum sepn., dispensing, etc. is thus eliminated and the method is ideally suitable for inspection of an emergency specimen.

Description

Detailed Description of the Invention The present invention is a novel 4f method for clinical chemical analysis of blood, urine, etc.
It concerns an analytical device.

Today, due to the widespread use of automated clinical chemistry analyzers, blood and urine tests are now only 9. ＜In the i time, go to the correct 1i1''
, and has made significant contributions in the clinical field.

However, in conventional clinical analysis, as shown in Figure 1, after the surface is taken (1), it is left to stand for a certain period of time and coagulated (2), and serum separation (3) is performed.
Through the process of dispensing, 1 liter of blood is separated, and in some cases, proteins are further removed, and then introduced into an analyzer for automatic analysis (4). This method requires a lot of time and effort.[Ill also uses a centrifugal force field to separate blood cells. There are blood cells at the bottom of the sample tube and white oil or blood in liquid form at the top, and the blood separated in this way is blue (also CL surface prefecture). Sucking in Bibetsu 1~'Nige'C'r) Trial with Japanese clothing! ! 'jl? 3. This kind of work is extremely troublesome, and there are many cases where urgent blood tests cannot be completed in time.

Therefore, the present invention eliminates and overcomes the above-mentioned conventional drawbacks (゛,
As shown by the dotted line at d3 in Figure 1, the purpose is to provide a new analytical device that can be analyzed immediately after blood collection, and does not require any work such as leaving it still, washing the surface, or using a minute hand. Rir.

The present invention (f4 or mono-substrate with porous thin dialysis I'
, ), the first chamber was filled with the sample to be analyzed ( ), and the second chamber was filled with the immobilized enzyme and the sample that had passed through the dialysis membrane. Extract the first liquid and/or inject a solution that reacts with the sample liquid,
A detection cell part is formed in the same substrate in communication with the second chamber, and the inside (reacted sample o'31
A solution is introduced into the detection region and a predetermined analysis is performed (7).

The invention will be explained below based on the embodiments shown in the drawings.

Figure 2 shows the main part of the tree R1 light! l Longitudinal cross-sectional view, Figure 3 is a △-Δ section view of Figure 2.
It is a base made of excellent glass, 1υreramikku, etc., and there are rooms 6, 7, and 8 inside.The space between the first room 6 and the second room 7 is Dialysis membrane 9
-C・Separated by -C, and the first part 1 contains the white liquid for inspection.
The liquid after inspection is discharged from the outlet 11.

The second chamber is filled with immobilized enzyme 12.
5. Dialysis II! ,! Trial pass 9! An enzymatic reaction occurs with one molecule (sugar). The second room contains dialyzed samples.
Extract the 31 liquid and/or perform a chemical reaction with the sample liquid 4z
It's easy! Liquid C is introduced through the inlet 13. Two or more types of the immobilized enzymes are packed in a mixed state, and both enzymes are used as a mediator to color the buffer solution.
I am working hard to ensure that i is carried out. The sample liquid that has developed one color or is in the process of developing color in the second chamber is introduced into the detection rail iji S through the passage 14 and kept in a stationary state. Above the detection cell section, one end of an A-boole fiber 15 is installed, and a light beam of a predetermined 11i wavelength is introduced from a light source 18 through a lens 16 and a filter 17 arranged at the width of the fiber. It is applied to the detection cell part through the base body 5. The light flux absorbed by the colored sample solution in the detection region further passes through the substrate)71
Detected by a photodetector 19 such as -1 to A-1.

Detection was completed in the test No. 11 solution) The first appearance I"20 was discarded into the drain tank (not shown). be done. Figure 4 shows the second
There is a schematic diagram C of the entire analyzer using the analytical device 8 shown in Fig. 1 and Fig. 3. Sample O'3i liquid is introduced and 110 is connected to the sample 1'81 syringe 21, and blood is directly supplied from the syringe. It is injected into the first chamber 6 of FIG. Introducing buffer solution
13 is connected to a C pump 23 via a switching valve 22, so that the buffer solution in the buffer solution 4iV 24 can be injected into the second chamber 7. 25 is a washing liquid tank C, in which 0.8% saline solution having the same salt concentration as in 1 is injected (for example, blood); , or through the valve 27 1) q test) 31 can be introduced into the liquid injector 21 . Also, the JJI ++ of No″Bice Δ
The waste liquid from 4020 is stored in the waste liquid tank 28. The output of the photodetector 19 is sent to the -C signal processing circuit 30 via the amplifier 29, and after being appropriately processed, it is sent to the recording or display device 3.
1, and the analysis (test) results are displayed.

I will explain the case in which a test for glucose in blood is carried out using an apparatus using the above-mentioned device.
As A, a porous polymeric material such as cellophane, vinyl, or polycarbonate having a thickness of several to several tens of microns is selected. Using such a dialysis membrane, an immobilized enzyme containing an appropriate mixture of glucose Axidis and Beroxidis was placed in the IX-divided room 7, and a pump 2 was installed in the room.
Introduce a buffer solution containing 4-21 antivirin by 3 and inject the sample (blood collected from syringe 21 into the first chamber) into the first chamber.

In such a situation, sugars in the J3-C1 plane, such as glucose, have a significantly smaller molecular weight than white spheres, so the glucose passes through the many small pores of the dialysis membrane 9 and becomes the C2
enters the room γ. This infiltrated glucose undergoes an enzymatic reaction with the immobilized enzyme charged with glucose, and is decomposed into gluconic acid and hydrogen peroxide. This hydrogen peroxide causes a chemical reaction with the 4)7-minoandipyrine in the chamber 7 using the Δxydis fixed in the chamber 7 as a catalyst, producing a C light color. The level of color development is 1α! : Since Li is proportional to Li in the dialysis membrane, a coloring phenomenon corresponding to the amount of glucose permeated from the dialysis membrane 519 can be obtained, and color measurement γ can be performed according to the amount of blood as a sample.

The infiltration, or dialysis, of molecules from the first chamber 6 to the second chamber 7 involves the arrival of one substance at the bottom, so that at the beginning 11 the second chamber contains only one molecule of glucose. Since the temperature is 0, glucose dialysis is performed very quickly. Then, as dialysis progresses, the target substance (glucose) that has entered reacts with the immobilized enzyme and its concentration decreases, so that the molecule progresses further in dialysis. However, the molecules that do not participate in the internal reaction of the dialyzed molecules have a concentration of 211
2, part 14, in a state where they are equal] ♂llr
Hassu(・tsubutsu).

In this way, the molecules that cover the purpose become extremely effective〕2
It is extracted into the chamber, and an elemental reaction occurs, and the coloring phenomenon progresses. After a preset period of 11.1 minutes has elapsed, the solution in the second chamber is pumped into the detection barrel 8 without operating the pump 23, and illuminated with a monochromatic light beam! Jll to the liquid J,
Measure the absorbance. The measurement results are recorded in the record t1 or on the display device 31. 1-J constant j, Il″A′
When the test 81 is completed, the changeover valve 22 is switched, the valve 27 is turned on, and the pump 2G is operated to pump the uC cleaning liquid into the first chamber 6 and the second chamber 7. This washing liquid C uses 0.8% saline solution (because it contains saline, it cleans the inside of each room without destroying blood cells).

In an actual device, after using this saline solution, it is recommended to wash with fresh water (J4).

When using an analytical device such as the one described above, +m
The process of JAt separation, dispensing, analysis (reaction) and measurement can be carried out in a single chip. ), making it ideal for testing emergency specimens.

But -6, the device t becomes a non-1:1 raw type and tries one]
The C' device, which does not require a sill 411 for moving liquids, buffers, etc., and requires 4τ piping, does not cause the problem of 111 purification-C/reli L1 scontamination.

The dialysis membrane 9, immobilized enzyme, and buffer are selected optimally depending on the type of protein to be tested.

FIG. 5 shows another embodiment of the invention;
This figure shows an example of the shape of a room. Figure J, clear 4
fJ, sea urchin, chambers 6 and 7 (6 is not shown in the figure) are made in a meandering shape so that the sample solution can efficiently penetrate into the 5A20 chamber and the permeated specimen 31 solution can effectively interact with the immobilized enzyme. It is designed to react. Of course, the shape shown in FIG. 5 is also an example, and this shape is not necessarily the best.

FIG. 6 shows still another embodiment of the present invention, in which the detection means is used instead of colorimetric measurement; . In the figure, the same symbols as in Figure 2 are the same 1! 1) In the process of the enzymatic reaction described above, arsenic is generated, and the peroxide During the reaction between hydrogen and the buffer solution, oxygen is released, which exists in the solution as dissolved acid.

When this dissolved oxygen is measured by the oxygen electrode 32, an enzymatic reaction occurs in the second chamber, and kC (kC) of sugar (11) can be actually measured.

Figure 7 shows I:J3 III of the analytical device according to the present invention.
IVI+ (・, multiple analytical devices △1.
△2, A3.

....... are connected to both columns, and different sugars (5+J/
, the molecular weight of each molecule) is measured sequentially using each device. That is, the sample liquid of the first device? ”] Input D 10 Sample entered from a (
Output from the outlet '11a (test of second device A2):
jl tl, the sample liquid enters the input Li 10b and enters the second device from the discharge port 11b to the third device △3
It is configured so that it enters the injection port 10G of. Each device also has a light source 18a, 181) as a measuring means.

18c ``'''''' and photodetectors 19a, 19b, 19
c.... is set (). Of course, depending on the pressing force of the immobilization element in each device, the type of buffer solution, the type and thickness of the dialysis membrane, etc. is used.

FIG. 8 shows still another example. In the example of FIG. 2, a plurality of immobilized enzymes were mixed and filled into the second chamber 7, but here two types of immobilized enzymes were mixed and filled ( 111), a third chamber 33 is formed between the second chamber and the detection cell section 8, and a third chamber 33 is formed between the second chamber and the detection cell section 8, The immobilized enzyme is filled with surrounding immobilized enzyme 34.

Then, between the second room 7 and the third section 1gi 33, the second room
A buffer solution inlet 35 is provided, and a buffer solution that is different from the buffer solution injected into the second chamber 7 is supplied. Of course, if necessary, a room filled with different types of immobilized enzymes can be provided.

With such a configuration, the first buffer injected from step 13 is the first buffer solution (7).The one that is most suitable for extracting only the desired sugar from the fit store is selected (7). ·tree,
A second buffer solution optimal for color reaction is added to the solution in which the sugars efficiently extracted from sea urchin-C react with the immobilized enzyme.
It is now possible to inject from the injection port 35, allowing efficient measurement.

FIG. 9 shows still another embodiment, in which an enzymatic reaction and a chemical reaction are combined. (P, >J-
A reaction test sample was injected from the human body (a chemical reaction was caused with the enzymatic reaction product produced in the second chamber), and the resulting color was measured by absorbance. .

In case of j, eel ii+i as explained above, the collected blood can be analyzed directly in the aliquot device C, and the blood serum can be analyzed by standing, coagulating or centrifuging as in f+L rice.
It eliminates the need for BLL, pipetting, etc., enables extremely rapid measurement, and is effective for testing urgent samples.

[Brief explanation of drawings]

Fig. 1 is a diagram showing the flow of conventional clinical chemistry analysis, Fig. 2 is a vertical sectional view of an analytical device showing an embodiment of the present invention, and Fig. 3
The figure is a sectional view taken along the line A-A in FIG. 2, FIG. 4 is a schematic diagram of an analyzer using the device shown in FIG. 2, and FIGS. d5 on the acceptance drawing. 5: M body 6: First chamber 7: Second chamber 8: Detection cell section 9: Dialysis membrane 10: Sample fluid injection 1] 11: Sample outlet 12: Immobilized enzyme 13: PAii liquid inlet 15 Nigel fiber 16: Optical lens 17: Filter 18: Light source 19: Photodetector Fig. 1 71 Fig. 4 Fig. 6 Fig. 7

Claims (1)

[Scope of Claims] (1) Single) "Separated by a thin porous dialysis membrane inside the body. Forming two rooms separated, the first room has 4J'
The second part is filled with immobilized enzyme, and the sample liquid that has passed through the dialysis membrane is extracted and/or a solution that reacts with the sample liquid is injected. A detection cell part is formed in the base body in communication with the second chamber, and a sample solution reacted in the second chamber is introduced into the detection cell part to perform a predetermined analysis. Analytical device for clinical analysis. 2) The analytical device for clinical analysis according to claim 1, wherein the second chamber is filled with a plurality of types of immobilized enzymes. (3) Form at least one chamber between the second chamber and the detection cell section, and fill the chamber with an enzyme different from the immobilized enzyme filled in the second chamber. An analytical device for clinical analysis according to claim 1. (4) An inlet for causing a chemical reaction with an enzyme reaction product between the second chamber and the detection cell section is provided, as set forth in Claim UIJJ Paragraph 1 or 1J Paragraph 2 iH. Analytical device for clinical analysis. (5) Sample) 3+liquid 1. discharged from the first chamber. L
Special H'1 introduced into the first chamber of another analytical device
An analytical device for clinical analysis according to any one of claims 1 to 4.