JPS5840100A - Measurement of uric acid and device therefor - Google Patents

Abstract

PURPOSE:The titled device that is composed of a combination of a means for causing an enzymatic reaction of uric acid, a means for converting carbonate ion or the like resulting from the enzymatic reaction into carbon dioxide by adding an acid, a means for absorbing the carbon dioxide in an absorbing solution and a means for detecting the change in the electroconductivity of the adsorbing solution. CONSTITUTION:A sample such as blood is injected from the sample injector 27 into the carrier to reach the immobilized uricase where the uric acid suffers enzymatic reaction to form carbon dioxide and hydrogen peroxide. At this time, the boric acid buffer solution is 8.5 in pH in the carrier, so that carbonic acid formed from carbon dioxide is in the form of bicarbonate ion and an acidic reagent is added from the inlet 28 to make the pH less than 1, thus most of the bicarbonate ion is changed into carbon dioxide gas. The uric acid in th sample is converted into carbon dioxide, carried into the second chamber 2 in the flow cell 1 to permeate through the membrane 2 in proportion to its partial pressure, into the first chamber 3 where the gas reacts with the absorbing solution. And the change in electroconductivity is detected by means of electrodes 7, 7' to determine the amount of the uric acid.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method and apparatus for measuring pb uric acid, which indirectly measures the concentration of uric acid (contained in a sample such as blood) from a conductivity change line of an absorption liquid.

In recent years, advances in biomedical measurements have enabled continuous remote measurement of physiological variables such as body temperature, gastrointestinal pressure, blood pressure, rate of respiration, and biological potential9, pO2, pCO2 in vivo. , blood pH and electrolytes,
In addition, the pH of the stomach was measured continuously. In this way, in the first part of the Jugyu medical measurement method, uricase enzyme is applied to uric acid in the sample to cause the fermentation reaction of formula (1), and the ultraviolet absorption of uric acid by uric acid is carried out. There is a so-called uricase purple absorption method that measures the concentration of uric acid in a subject from changes in Z).

Neu'Jfjt+ 2u2o+02 -, :1
-Saw meter---+co242■2o2-1-(1) However, in the spectrophotometric method such as the above-mentioned uricase ultraviolet absorption method, if the subject contains a direct protein, etc., the protein is detected. There is a drawback that it is impossible to measure total +ru when blood is the subject because stool must be removed. 1. It also had the disadvantage that it took a long time to develop the color, making it impossible to quickly measure uric acid in the sample.

The present invention was made in view of these drawbacks, and its purpose is to quickly and accurately remove uric acid contained in a specimen such as a whole surface, without compromising pretreatment operations such as protein removal. How to measure uric acid 41+11'ij
? dl,, IE? 1h1ij (surcotonial.

Hereinafter, the present invention i+r will be explained in detail using the drawings.

FIG. 1 is a configuration explanatory diagram showing an embodiment of the present invention, and in the figure, a 1i':l flow cell is made of a thin film such as r'TFE, silicone rubber, etc., which does not allow ions to pass through but allows gas to pass through. A first chamber 3 and a second chamber 4 are adjacent to each other with a diaphragm 2 in between.

1'i&, the first room 3 is provided with a t, l introduction 116 and an outlet 51, and the m2 chamber 4 is provided with an l] introduction 116 and an egress 16I. Furthermore, 2] J:I, for example, a solution containing borate buffer, NaCe1 and E'l' 1M andenite aromatic acid, etc.
Inlet where carriers consisting of deaf people are introduced, 2: (r, J
A sample injector 27 and a support such as glass beads or nylon tubes are provided at the outlet of the carrier, which injects the specimen into the flow path from the introduction 1] 21 to the introduction 16 of the second housing 4. A single fermenter 22 in which uricase solid enzyme is immobilized is provided. In addition, in the middle of the flow path from the immobilized enzyme 22 to the inlet 6 of the second tube 4, for example, IIce or iNo. A reagent addition port 28 is provided for adding an acidic reagent such as an IN solution. On the other hand, 24 is an absorption liquid introduction port, and 1 and the collected liquid 26 in the container 25 (for example, 0.00
It is installed so that it can aspirate an alkaline solution (such as 2H KOII). 10 is an outlet of the absorbent liquid 1, and the absorbent liquid is output from the outlet 5 of the first chamber 3.
0117: In the middle of the flow path, for example, there is a power supply 8 and a galvanometer □
For example, the electrodes 7, 7I with which a total of 9 are connected have an inner diameter of 10 zoI.
11, and is provided near the outlet r15' to detect the electrical conductivity of the absorption liquid drawn out from the outlet 5I.

-1- In the embodiment of the present invention consisting of the configuration described above, introduction 1
] 21 flows through the immobilized enzyme 22, the introduction port 16, the second chamber 4, and the outlet port 6I, and is discharged from the outlet port 23. Also, the carrier introduced from the inlet port 24 2G flows red through the inlet 5, the first chamber 3, and the outlet 5, and is discharged from the outlet 10.

Therefore, when the specimen is injected into the gearia from the sample injector 27, the specimen is carried by the carrier and reaches the immobilized enzyme 22, where it undergoes an enzymatic reaction as shown in equation (1) above and becomes C02. and lI202. At this time, since the boric acid buffer in the gearia has a pH+ of 845,
The carbonic acid produced from the above CO is bicarbonate ion llCO3
-. When the acidic reagent is added into the gearia from the reagent addition port 28 and the plf of the mixed solution consisting of Ki, Yria, etc. becomes 1 or less, the bicarbonate ion 1■C
Most of O3- will be 002 swords. In this way, the uric acid contained in the subject is converted into CO2 gas, which is transported by the gear from the introduction 1'16 to the 882 chamber 4 in the 70-cell 1. In the second chamber 4, the partial pressure of the 002 gas in the gear 1 is proportional to 0.11, and the diaphragm 2 passes through the diaphragm 17 to reach the first palm 6. In the first chamber 6, -F formula (2) Reacts with the absorbing liquid like this. 1
Due to this reaction, the absorption rate - lead 11f1 rate changes, and the lead 〒
IY, rate co→-2Igu011--neeee-6ji, ni2C03-II+20------------,-(
2) The change is detected by ``Niki Dentsubaki 7.7+''. In this way, the uric acid in the subject is determined indirectly from the conductivity and rate change detected by the electrodes 7 and 71 through predetermined signal processing (y1 not shown).

Also, FIG. 2 shows an embodiment of the present invention. FIG. 3 is an exploded perspective view of the norocell shown in FIG. 3; FIG. In Figures 2 and 5 "C1i, 1.1.7i"
J block, each introduced l',,l 1.1. a
, 1.7n and leads 1111b, 17b are provided, and a cavity 1.3.1.6 forming the first and second chambers, respectively, is provided with 1.2.15 rJ and a gasket l. Also, 14 is a diaphragm that does not allow ions to pass through but allows gas to pass through. -1 In the 1-Hill consisting of two configurations, the carrier containing the analyte etc. is introduced from the introduction II 1.7a, passes through the cavity 16 and is led out from the outlet [1171], and the absorption liquid is introduced from the introduction port 11a. The sound is introduced through the air 1lBI sound 1s13 and is led out from the outlet 111).

1, CO contained in the carrier flowing through the cavity 16
The gases t and l pass through the diaphragm 14, reach the cavity 13, and react with the absorption liquid flowing through the cavity 1:I.

X in the embodiment of the present invention as described in detail in 1-1 below;
1. Requires pre-treatment operations such as drying, sludge removal, and rinsing.
It quickly and accurately detects uric acid contained in the sample.
0]! It has the advantage of being able to

In IJi, FIG. 4 shows a configuration 1 showing another embodiment of the present invention.
This is a clear diagram of 191, and in the diagram, 9 indicates the absorption liquid inlet, and 30
This is a sample switching pulp that supplies the analyte injected from the Keda/Pull injector 27 into the thick gearia by switching the flow path. In figure 4, it is the same as in figure 1.
Numerical symbols (for example, fi'a, 5'b are the same numerical symbols as 51) are used once to convey their meaning, and redundant explanations will be omitted here. In another embodiment of the present invention based on the above configuration, a part of the carrier introduced from the introduction 121 is transferred to the second chamber 4as of the flow cell 1a and the second chamber 4as of the flow cell 1a and the outlet 1''6'a%. via outlet 2
3a, and the remaining +J-immobilized enzyme 22,
4 through the inlet 6b, the second chamber 4b of the flow cell 11), and the lead-out D6Ib].

Moreover, the absorption liquid 26 in the container 25 is introduced from the absorption liquid introduction port 24, and the absorption liquid 26 is introduced from the absorption liquid introduction port 24, and the absorption liquid 26 is introduced from the introduction port 5b,)] -1!
11), and the outflow ITI 5'l)
and is discharged from the discharge port 10b,
The other part 0 is introduced from the absorption liquid introduction port 29, and the introduction 17j
5+1, the first chamber 3a of the flow cell 1a, and the outflow [
215 ma is threaded through the thread Y, ha, J) + is discharged from the outlet 10a. By the way, when a predetermined amount of the analyte injected from the sample injector 27 is supplied to the flow cell 1a through the sample switching pulp 30, the analyte cannot be carried by the carrier.
However, when the acidic reagent is added from Reagent Addition II 2B11, if carbonate ion and bicarbonate ions are included in the above-mentioned sample, these ions are converted to CO2 gas. After that, the CO2 gas and the specimen are transported to the second chamber 4a in the flow cell-a by the gear rear, and the
Only 02 gas is proportional to the distribution 1. The liquid then passes through the partition IIφ2a and reaches the first chamber 3a, where it reacts with the absorption liquid as shown in equation (2) above. Due to this reaction, the conductivity of the absorption liquid in the first I<3+ changes, and this change in conductivity causes the electrodes 7n, 7'a
detected by. In addition, the second chamber 4 of the flow cell 1a
After carbonate ions and bicarbonate ions are removed from the specimen 1 in a as described above, it is transported again to the carrier and discharged from the discharge ITI 23a via the outlet 6'a. On the other hand, when a predetermined amount of the analyte introduced from Zampurui 7 and Kuta 27 is supplied to the second carrier channel leading to the immobilized enzyme 22 via the sample switching pulp 30, the analyte 1 ′: [immobilized enzyme 22 carried by carrier
The CO
2 and lI202. In this case, the boric acid buffer in the carrier l', plf is 8.5, so it is -1
-carbonate 0 bicarbonate ion lIc0 produced from CO2
-. Further, when an acidic reagent is added into the carrier from the reagent addition port 281 and the pII of the mixed solution consisting of the carrier etc. becomes 1 or less, the bicarbonate ion 1C
The thick part 6) of O3- reaches the C02 gas. Therefore, the uric acid contained in the sample is converted to CO2 gas,
v The IJ is carried from the inlet 6b to the second chamber 4b in the flow cell 1b.

In the second chamber 4b, C02 gas permeates through the diaphragm 2 in proportion to the partial pressure of the C02 gas in the gear rear, and the first chamber 31) reaches K'C.
It reacts with the absorption liquid as shown in equation (2) above. Due to this reaction, the conductivity of the absorption liquid in the first chamber 3b changes 17, and this change in conductivity is detected by the electrodes 7b and 71b. Therefore, a predetermined calculation process (Fig. (not shown) is applied, even if carbonate ions or bicarbonate ions are contained in the carrier or the sample, uric acid in the sample will be measured regardless of these ions.

According to other embodiments of the present invention as described in detail above, even if carbonate ions or 11 carbonate ions are rarely contained in the specimen, carbonate ions or bicarbonate ions are present in the carrier. Zl has the advantage of being able to accurately measure uric acid in a test sample quickly and without being interfered with by C), even if it is dissolved.

[Brief explanation of the drawing]

11, a configuration explanatory diagram showing an embodiment of the present invention, and 2.
6 and 6 are an exploded slanted view and a partially assembled view of a flow cell according to an embodiment of the present invention, and FIG. 4 is a configuration explanatory diagram showing another embodiment of the present invention. 1.1a, 1+J...flow cell, 2.2a, 2+
)-diaphragm, 3.3a. 3L+-1st character, 4.4a, Sen...2nd house, 5.5a
, 5b, 6.6a. Gb, 21. , 24.29・”Introduction [-1,5',
5'a, 5'b, 6', 6'a,, 6'b...
・Derivation [1,? , 7n, 7b, 7°, 7'a,
7'b -1 [Polar, 10.10a. 10b, 23...Discharge port, 25...Response, 2
6...Absorption liquid, 27...-Lean pull injector, 28.28a, 28b...Reagent addition port, 30...
・Rimple switching pulp. Stone 3 呵/ltb

Claims (3)

[Claims] (1) A means for causing an enzymatic reaction by causing uricase to act on uric acid contained in the specimen, and a means for adding an acid to convert carbonate ions or bicarbonate ions generated in the enzymatic reaction into carbon dioxide gas. means for absorbing the carbon dioxide gas into the basic absorption liquid by permeating the carbon dioxide gas through a diaphragm that does not allow ions to pass through, and means for detecting the amount of change in conductivity of the basic absorption liquid. A method for measuring uric acid, characterized in that uric acid in a subject is indirectly measured from a change in the conductivity of the absorption liquid. (2) A first port is provided with an inlet and a lead-out port 1 through which a basic absorption liquid that absorbs carbon dioxide gas is introduced and taken out, respectively, and constitutes a flow path on one side of the flow cell, and a carrier containing an analyte is provided. A second chamber is provided with an inlet and an outlet for introduction and extraction, respectively, and is separated from the first chamber by a diaphragm that does not allow dissolved ions to pass through, but allows gas to pass through, and constitutes a flow path on the other side within the flow cell. a chamber, an immobilized enzyme on which a uricase enzyme is immobilized that acts on uric acid in the specimen to cause an enzymatic reaction, and a flow path from the immobilized enzyme to the inlet of the second chamber. A reagent addition port provided in the
A sample injector is provided near the inlet of the immobilized enzyme and injects the sample from a predetermined side port, and an electrode is provided near the outlet of the first chamber and detects the conductivity of the absorption liquid. l/, a uric acid measuring device that indirectly measures uric acid in a subject from the amount of change in conductivity of the absorption liquid. (3) An inlet and an outlet are provided through which a basic absorption liquid that absorbs carbon dioxide gas is introduced and extracted, respectively.
In a first chamber constituting a flow path on one side of the cell, an electrode provided near an outlet of the first chamber to detect the conductivity of the absorption liquid and a carrier containing an analyte are introduced and extracted, respectively. A second chamber is provided with an inlet and an outlet, and is separated from the first chamber via a diaphragm that does not allow ions to pass through but allows gas to pass through, and constitutes a flow path on the other side within the flow cell; The introduction of two chambers [1] and the reagent addition provided near 1 are placed in the -IM of the immobilized enzyme, and the immobilized enzyme is injected from the sample injector into the second and [streams: l , -tJ Uric acid 4 (11
Fixed device a'.