JPS58131546A - Apparatus for minute flow analysis - Google Patents

Abstract

PURPOSE:To decrease the using quantity to a component of be inspected and reagent, by supplying the component to be inspected and the reagent for an optical analysis in a single pipe for transfer liquid supply through juxtaposed plural sleeve flow pipes and mixing by making it into a turbulent flow and then, supplying said mixture to a flow cell. CONSTITUTION:A sample such as blood serum introduced from a sample introducing entrance 5 is supplied to a fixing enzyme column 6 together with a buffer 7 and said sample is introduced into a single pipe 1 from a supply pipe 11 through a sleeve flow pipe 2a as a component to be inspected containing hydrogen peroxide. Besides, a luminol solution 8 as a luminous reagent and a potassium ferricyanide solution 9, are introduced into the tube 1 from each supply pipe 12, 13 through sleeve flow tubes 2b, 2c and continuously, this sleeve-like flow is introduced into a slender pipe part 15. Then, said flow is broken by making into a turbulent flow by passing through a filter 3 and the component to be inspected is mixed thoroughly with the luminous reagent. This mixed solution is supplied to a flow cell 41 and luminous quantity of the mixed solution passing through the cell 41, is detected by a light detection part 4 including a photodetector 42 and the concentration of glucose contained in the trace quantity of a sample is determined quantitatively.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an IkIk flow analysis device. Zara 4
C In detail, compared to the conventional 70-analyzer, the test component (
The present invention relates to a microflow analyzer that can significantly reduce the amount of samples (sample) and reagents used for optical analysis, and that can provide stable optical detection signals.

Conventionally, flow analysis methods based on optical analysis have been used in various fields such as general analysis and clinical analysis, and because continuous reagents are supplied continuously, there are interlayer points that require a large amount of reagents. there were. Furthermore, in order to perform multi-item analysis using limited samples such as biological samples, it has been desired to obtain more accurate analytical values using as small a sample as possible.

In this regard, a flow system is used to rapidly inject a small amount of sample and optical analysis reagent into a measurement cell, and then the flow is temporarily stopped and analysis is performed based on changes in luminescence, absorption, or fluorescence intensity in the measurement cell over time. The stopped-arrow method has been proposed, but it is difficult to obtain a stable photodetection signal.

This invention has been made in view of the above points.Thus, according to this invention, a test component and a reagent for optical analysis are respectively flowed through a plurality of sheath flow tubes arranged in parallel within a single tube for supplying a transfer liquid. a mixing means connected to the supply means for turbulently mixing the plurality of sheath flows; and a mixing means for continuously detecting the luminescence, absorption or fluorescence intensity of the mixed liquid from the mixing means. In accordance with the present invention, there is provided a micro flow analysis device comprising a photodetecting section, in which various reagents such as luminescent, coloring, or fluorescent reagents can be mentioned as reagents for photoanalysis.

Hereinafter, the present invention will be described in detail with reference to the drawings.

FIG. 1 is a schematic diagram showing a blood glucose concentration measuring device which is an example of the micro flow analyzer of the present invention. Inside the single tube (1), there are midstream tubes (21) for test components, which are arranged parallel to each other in the tube axis direction, and sheath flow tubes (2b) and (2b) for luminol solution and red dish salt solution, which are luminescent reagents. 2C). 2, sheath flow tube (2
a) is connected to a test component supply pipe provided with a sample inlet +51 and an immobilized enzyme column (61 on which glucose okindase is immobilized) in one stage of Ai. (A test component containing hydrogen peroxide produced corresponding to the amount of glucose in the sample by supplying the sample @ (serum etc.) introduced from 51 with buffer (7) to the immobilized enzyme column +61 with a pump A single tube (1
) is configured to be fed as a pod stream within a pod. Also,
The sheath flow tubes (2b) and (2C) are connected to the luminol solution supply pipe flffi and the red blood salt solution supply pipe a3, respectively, and pump a predetermined amount of the luminol solution (marked) and the red blood salt solution (9). It is arranged to feed each of them as an intermediate stream into the single pipe (1) via the respective pipes (1).

On the other hand, the other most characteristic mixing means of this invention consists of a filter (3) provided in the mfse- through the squeeze pipe @4 in the single pipe (1).

The thin tube part that has passed through the filter (31) is connected to the flow 7/I/iia in the form of a spiral flow path in the same plane, and digitally integrates the amount of received light with a photoreceiver consisting of two cells←υ and a photomultiplier tube. Light detection unit 14 due to photon counter fight
1 is configured, 2, - indicates a recorder, and αG indicates a waste liquid reservoir, respectively.

In the above configuration 2, the test component containing hydrogen peroxide supplied from the supply pipe bend is blown into a single tone by the sheath flow 11 (2 m).
It is introduced as Saya-ryu within the country. In addition, luminol solution ta+ and red blood salt solution (9) as luminescent reagents are also passed through the supply tubes (I21 and a), respectively, through the sheath flow tubes (2b) and (2).
C) is introduced as a sheath flow within a single if tll (each sheath flow is shown by a broken line in Fig. 2).Subsequently,
These sheath flows are guided by the Ml buds and flow into the filter (3) while maintaining a bundle shape close to each other. By passing through the filter (31), each sheath flow becomes turbulent and destroyed, resulting in efficient mixing of the analyte component and the luminescent reagent.

The mixed liquid that has passed through the mixing means is then supplied to the flow cell from the inlet IA1 and discharged from the drain (B).
During this time, the amount of light emitted from the mixed liquid passing through 70-7V is integrated over a certain period of time using a photodetector hook and a photon counter, and by comparing it with a standard sample, the hydrogen peroxide concentration in the test component can be determined.
As a result, the glucose concentration in a trace sample can be determined with high sensitivity.

2. If the flow rate of the transfer liquid (buffer: sodium carbonate/sodium hydrogen carbonate system, PH1O,o) in the single tube (1) is 1 m/5 ec (2 ωm in terms of tube diameter), the flow rate of the test component is Approximately 0.3 μt/sec is appropriate, and the flow rates of luminescent reagents +81 and 191 are approximately 1.6 μL/s each.
e c is appropriate. Roughly speaking, the diameter of the sheath flow tube (2a) is approximately 20 pm, and the diameter of the sheath flow tube (2b), (2c) is approximately 20 pm.
X) tl[yk are each approximately 45 μm, which is suitable for forming a sheath flow. Therefore, even if monitoring is performed for 10 seconds, for example, the required amount of the test component is 3 μt, the consumption of the crack sample is very small, and the required amount of the luminescent reagent is 1 μt.
It will be seen that at 6 μt, the amount is significantly reduced compared to the conventional one.

As the &t filter (3), for example, a porous glass filter is suitable. The micro flow analyzer of the present invention is not limited to the specific examples described above. For example, a structure based on a commonly used spectrophotometry method or a structure based on a fluorescence method may be applied. Instead of the luminescent reagent in the example, a coloring reagent or a fluorescent reagent is supplied as a sheath flow in a single tube, and an absorptiometer or fluorometer equipped with a known flow system is used as the photodetector.

The sample inlet is then directly connected to the sheath flow tube via a pump or diluter. As the means for supplying the sheath flow, any known sheath flow generating means in the field of hemocytometers and the like can be applied.

Further, in the present invention, the number of sheath flow tubes arranged in parallel is not limited to three, but is determined as appropriate depending on the analytical method to be applied. Further, the mixing means may be of any type as long as it can at least make the sheath flow turbulent and mix it, and in addition to the filter, for example, a spiral pipe can be used.

Na S? sAs a special application, I in a single pipe as shown in Fig. 2
By applying a structure in which a light receiver is arranged or moved along the flow path of the IjAw section, the progress of the reaction can be confirmed, which is very effective for monitoring enzyme reactions, for example.

As mentioned above, the micro flow analyzer of this invention can be used for flow analysis. Since both the sample and reagent can be used in very small amounts, it is especially useful as an analyzer for biological samples. Because it uses a coarse flow and its turbulent mixing means, excellent mixing is achieved, and even though a small amount of sample or reagent is used, optical detection is more stable than the stop-draw method. get a signal

[Brief explanation of drawings]

FIG. 1 is a schematic diagram showing a blood glucose concentration measuring device which is an example of the subtle flow analyzer of the present invention, FIG. 2 is an enlarged view of section P in FIG. 1, and FIG. It is a top view which shows 70-cells of a spiral flow path shape in the same plane. fi+...Single tube for supplying liquid to be transferred, (2a), (2b),
(2c) ... sheath flow tube, (3) ... filter,
4)...Photodetection section.

Claims (1)

[Claims] 1. A supply means for supplying a test component and a reagent for optical analysis as intermediate flows through a plurality of sheath flow tubes arranged in parallel in a single pipe for supplying the transfer liquid, and a supply means connected to the supply means to disturb the plurality of sheath flows. A trace amount analysis device 70, comprising a mixing means for fluidizing and mixing, and a light detection section capable of continuously detecting the luminescence, absorption, or fluorescence intensity of the mixed liquid from the mixing means.