JPS6334429B2 - - Google Patents
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- Publication number
- JPS6334429B2 JPS6334429B2 JP56163590A JP16359081A JPS6334429B2 JP S6334429 B2 JPS6334429 B2 JP S6334429B2 JP 56163590 A JP56163590 A JP 56163590A JP 16359081 A JP16359081 A JP 16359081A JP S6334429 B2 JPS6334429 B2 JP S6334429B2
- Authority
- JP
- Japan
- Prior art keywords
- liquid
- urea
- measured
- chamber
- conductivity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
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- 239000007788 liquid Substances 0.000 claims description 77
- 238000010521 absorption reaction Methods 0.000 claims description 32
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 25
- 239000004202 carbamide Substances 0.000 claims description 25
- 108010093096 Immobilized Enzymes Proteins 0.000 claims description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 108010046334 Urease Proteins 0.000 claims description 7
- 238000006911 enzymatic reaction Methods 0.000 claims description 7
- 239000012528 membrane Substances 0.000 claims description 7
- 239000003957 anion exchange resin Substances 0.000 claims description 4
- 239000007789 gas Substances 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 6
- 238000005342 ion exchange Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 238000000691 measurement method Methods 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 239000003929 acidic solution Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/58—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving urea or urease
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Description
【発明の詳細な説明】
本発明は、血液等の被測定液に含まれている尿
素の濃度を吸収液の導電率変化量から間接的に測
定する尿素の測定方法および測定装置に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method and apparatus for measuring urea that indirectly measures the concentration of urea contained in a liquid to be measured such as blood from the amount of change in conductivity of an absorbing liquid.
近年、生医学的測定法の発展により体温、胃腸
内の圧力、血圧、呼吸の速度、および生物学的ポ
テンシヤルのような生理学的変数を連続的に遠隔
測定したり、生成内におけるPO2、PCO3、血液
のPHおよび電解質、並びに胃のPHを連続的に測定
したりする測定方法や測定装置等が開発されるよ
うになつた。 In recent years, advances in biomedical measurements have enabled the continuous telemetry of physiological variables such as body temperature, gastrointestinal pressure, blood pressure, rate of respiration, and biological potentials, as well as the production of PO 2 , PCO 3. Measuring methods and devices for continuously measuring blood PH and electrolytes as well as gastric PH have been developed.
上記生医学的測定法の一つとして、被測定液中
の尿素分解酵素であるウレアーゼ酵素を作用させ
て下式(1)の酵素反応を生じさせ、生成したNH3
をNH3ガス電極若しくはNH4 +カチオン電極にて
検出することにより間接的に被測定液中の尿素を
測定する方法がある。 As one of the above-mentioned biomedical measurement methods, urease enzyme, which is a urea-degrading enzyme, is activated in the liquid to be measured to cause the enzymatic reaction of the following formula (1), and the generated NH 3
There is a method of indirectly measuring urea in a liquid to be measured by detecting it with an NH 3 gas electrode or an NH 4 + cation electrode.
然しながら、上記尿素測定方法においては、上
記電極からの出力信号とNH3濃度との関係は下
式(2)のようなネルンストの式に従つて指数関数的
関係にあり、比較電極の液間電位差の微少変化が
NH3濃度の測定に際して大きな誤差要因になる
といつた欠点があつた。 However, in the above urea measurement method, the relationship between the output signal from the electrode and the NH 3 concentration is an exponential relationship according to the Nernst equation as shown in equation (2) below, and the liquid junction potential difference of the reference electrode The slight change in
It had a drawback that it was said to be a major source of error when measuring NH 3 concentration.
Ｅ＝Eo＋2.303RT／Ｆ（log［NH3］＋log ａ）
……(2)
〔log［NH3］＋log ａ＝log（［NH3］xa）
＝1og［OH-］
［OH-］／［NH3］＝ａ
Ｅ：平衡電極電位、Eo：標準電極電位、Ｒ：ガ
ス定数、Ｔ：絶体温度、Ｆ：フアラデー定数、
〔NH3〕：NH3濃度、〔OH-〕：OH-濃度、ａ：定
数〕
また、本願発明の発明者らは上記欠点を除去す
るため次のような尿素測定方法および測定装置を
発明し、本願発明と同一出願人名ですでに出願済
みである（昭和55年11月25日出願、特願昭55−
165597、「尿素の測定方法および測定装置」、以下
「前出願」という）。該前出願の要旨は以下の(1)〜
(3)項に示す如くである。すなわち、
(1) 被測定液に含まれている尿素にウレアーゼを
作用させて酵素反応を生じさせる手段と、被測
定液に試薬を添加してアルカリ性にすることに
より前記酵素反応で生成したNH4 +若しくは
NH3をNH3ガスに変化させる手段と、該NH3
ガスをNH3ガス透過性膜を透過させることに
より酸性若しくは弱酸性の溶液からなる吸収液
と接触させて反応させる手段と、該反応による
前記吸収液の導電率変化量を電源と検流計が電
気的に接続された電極を用いて検出する手段と
を講じて、前記吸収液の導電率変化量から間接
的に被測定液中の尿素を測定することを特徴と
する尿素の測定方法。E=Eo+2.303RT/F(log[NH 3 ]+log a) ...(2) [log[NH 3 ]+log a=log([NH 3 ]xa) =1og[OH - ] [OH - ]/[ NH 3 ]=a E: equilibrium electrode potential, Eo: standard electrode potential, R: gas constant, T: absolute temperature, F: Faraday constant,
[NH 3 ]: NH 3 concentration, [OH - ]: OH - concentration, a: constant] In addition, the inventors of the present invention invented the following urea measuring method and measuring device in order to eliminate the above drawbacks. , has already been filed under the same applicant's name as the claimed invention (filed on November 25, 1982, patent application 1982-
165597, “Method and device for measuring urea” (hereinafter referred to as “previous application”). The gist of the previous application is as follows (1) ~
As shown in paragraph (3). That is, (1) a means for causing an enzymatic reaction by causing urease to act on urea contained in a liquid to be measured, and a means for causing an enzymatic reaction by adding a reagent to the liquid to be measured to make it alkaline. + or
A means for converting NH 3 into NH 3 gas, and the NH 3
A means for causing the gas to contact and react with an absorbing liquid made of an acidic or weakly acidic solution by passing through an NH 3 gas permeable membrane, and a power source and a galvanometer to measure the change in conductivity of the absorbing liquid due to the reaction. 1. A method for measuring urea, comprising: detecting using electrically connected electrodes to indirectly measure urea in a liquid to be measured from the amount of change in conductivity of the absorption liquid.
(2) NH3ガスを吸収して反応する吸収液が流入
口から導入されるとともに流出口から流出され
フローセル内の一側の流路を構成する第１室
と、被測定液が流入口から導入されるとともに
流出口から流出されフローセル内の他側の流路
を構成するとともに前記第１室とはNH3ガス
透過性膜を介して隣接する第２室と、被測定液
中の尿素に作用して酵素反応を生ぜしめるウレ
アーゼ酵素が固定化された固定酵素と、該固定
化酵素から前記第２室の流入口に至る流路の途
中に設けられた試薬添加口と、前記第１室の流
出口よりも下流の流路に設けられるとともに電
源および検流計と電気的に接続され前記吸収液
の導電率を検出する電極とを具備し、吸収液の
導電率変化量を計測して被測定液中の尿素を間
接的に定量することを特徴とする尿素測定装
置。(2) A first chamber in which an absorption liquid that absorbs and reacts with NH 3 gas is introduced from the inlet and flows out from the outlet, forming a channel on one side of the flow cell, and a liquid to be measured is introduced from the inlet. The urea in the liquid to be measured is introduced and flows out from the outlet, forming a flow path on the other side of the flow cell. an immobilized enzyme on which a urease enzyme is immobilized that acts to cause an enzymatic reaction; a reagent addition port provided in the middle of a flow path from the immobilized enzyme to the inlet of the second chamber; and an electrode that is provided in a flow path downstream of the outflow port and is electrically connected to a power source and a galvanometer to detect the conductivity of the absorption liquid, and measures the amount of change in the conductivity of the absorption liquid. A urea measuring device characterized by indirectly quantifying urea in a liquid to be measured.
(3) NH3ガスを吸収して反応する吸収液が流入
口から導入されるとともに流出口から流出され
フローセル内の一側の流路を構成する第１室
と、被測定液が流入口から導入されるとともに
流出口から流出されフローセル内の他側の流路
を構成するとともに前記第１室とはNH3ガス
透過性膜を介して隣接する第２室と、前記第１
室の流出口よりも下流に設けられるとともに電
源および検流計と電気的に接続され前記吸収液
の導電率を検出する電極とを、被測定液が流れ
る流路において固定化酵素の上流に付加して配
設し、固定化酵素の上流および下流における２
つの電極で検出された前記吸収液の導電率変化
量の差から被測定液中の尿素を定量することを
特徴とする上記第(2)項記載の尿素測定装置。(3) A first chamber in which an absorption liquid that absorbs and reacts with NH 3 gas is introduced from the inlet and flows out from the outlet, forming a channel on one side of the flow cell, and a liquid to be measured is introduced from the inlet. The first chamber is introduced and flows out from the outlet to form a flow path on the other side of the flow cell.
An electrode is provided downstream of the outlet of the chamber and electrically connected to a power source and a galvanometer to detect the conductivity of the absorption liquid, and is added upstream of the immobilized enzyme in the flow path through which the liquid to be measured flows. 2 upstream and downstream of the immobilized enzyme.
The urea measurement device according to item (2) above, wherein urea in the liquid to be measured is determined from a difference in the amount of change in conductivity of the absorption liquid detected by two electrodes.
然し乍ら、前出願の発明においては、吸収液
（例えばHCl）とNH3が反応して生ずる物質（例
えばNH4Cl）の導電率に比して吸収液の導電率
は一般に大きいため、該吸収液導電率をベースと
して検出する電極の出力信号はＳ／Ｎ比が悪くな
り易いという欠点があつた。また、吸収液の温度
が変化すると、該吸収液はHCl等でなるため該吸
収液の導電率は温度変化に伴なつて変化し易いと
いう欠点もあつた。 However, in the invention of the previous application, since the conductivity of the absorbing liquid is generally higher than that of the substance (e.g. NH 4 Cl) produced by the reaction between the absorbing liquid (e.g. HCl) and NH 3 , the absorbing liquid The output signal of the electrode, which is detected based on conductivity, has a drawback that the S/N ratio tends to be poor. Another disadvantage is that when the temperature of the absorbing liquid changes, the conductivity of the absorbing liquid tends to change as the absorbing liquid is made of HCl or the like.
本発明は、前記従来例および前出願のかかる欠
点に鑑みてなされたものであり、その目的は、上
述のような欠点が全て除去され被測定液中の尿素
が正確に測定できるような尿素の測定方法および
測定装置を提供することにある。 The present invention has been made in view of the drawbacks of the conventional example and the previous application, and its purpose is to eliminate all of the above-mentioned drawbacks and to provide a urea solution that can accurately measure urea in a liquid to be measured. The object of the present invention is to provide a measuring method and a measuring device.
以下、本発明について図を用いて詳細に説明す
る。図は、本発明の実施例を示す構成説明図であ
り、図中、１はフローセルであつて、NH3ガス
透過性膜２を介して第１室３と第２室４が隣接し
ている。また、第１室３には流入口５と流出口
５′が設けられ、第２室４には流入口６と流出口
６′が設けられている。更に、１１は被測定液導
入口、１３は被測定液排出口であつて、被測定液
導入口１１から第２室４の流入口６に至る流路の
途中にはウレアーゼ等の酵素が固定化された固定
化酵素１２および被測定液に試薬を添加するため
の試薬添加口１７が設けられている。更にまた、
１４は吸収液導入口であつて、容器１５内の吸収
液１６（例えば1/100ＮのHCl）を吸引できるよ
うに設置されている。また、７は流出口５′の下
流に配設され陰イオン交換樹脂が充填されてなる
カラム、１０は吸収液排出口である。更に、カラ
ム７から吸収液排出口１０に至る流路の途中に
は、電源９と検流計９′が電気的に接続された電
極８，８′が例えば内径約１mmの配管に装着され
カラム７の出口の近くに設けられている。尚、固
定化酵素１２はクロマトグラフ用カラムに酸素を
担体に固定化し充填されたもの若しくは酵素固定
膜をNH3ガス透過性膜２と貼合わせたもので代
替することも可能である。 Hereinafter, the present invention will be explained in detail using figures. The figure is a configuration explanatory diagram showing an embodiment of the present invention. In the figure, 1 is a flow cell, and a first chamber 3 and a second chamber 4 are adjacent to each other with an NH 3 gas permeable membrane 2 in between. . Further, the first chamber 3 is provided with an inlet 5 and an outlet 5', and the second chamber 4 is provided with an inlet 6 and an outlet 6'. Furthermore, 11 is a sample liquid inlet, 13 is a sample liquid outlet, and an enzyme such as urease is fixed in the flow path from the sample liquid inlet 11 to the inlet 6 of the second chamber 4. A reagent addition port 17 is provided for adding a reagent to the immobilized enzyme 12 and the liquid to be measured. Furthermore,
Reference numeral 14 denotes an absorption liquid introduction port, which is installed so as to be able to suck in the absorption liquid 16 (for example, 1/100N HCl) in the container 15. Further, 7 is a column disposed downstream of the outlet 5' and filled with an anion exchange resin, and 10 is an absorption liquid outlet. Furthermore, in the middle of the flow path from the column 7 to the absorption liquid outlet 10, electrodes 8 and 8', to which a power source 9 and a galvanometer 9' are electrically connected, are attached to, for example, piping with an inner diameter of about 1 mm. It is located near exit 7. The immobilized enzyme 12 can also be replaced by a chromatography column filled with oxygen immobilized on a carrier, or a chromatography column in which an enzyme-immobilized membrane is bonded to the NH 3 gas-permeable membrane 2.
以下、本発明実施例の動作について説明する。
図において被測定液導入口１１から導入された被
測定液は、固定化酵素１２を通り、被測定液中の
尿素がウレアーゼの作用によつて前式(1)のような
酵素反応をうける。その後、該被測定液に試薬添
加口１７からNaOH等の試薬が添加されて被測
定液がアルカリ性となり、前式(1)で生じたNH3
やNH4 +がNH3ガスに変化する。而して、該被測
定液は流入口６から第２室４へ至り、流出口６′
からフローセル１の外へ流出し、被測定液排出口
１３から外部へ排出される。一方、吸収液導入口
１４から導入された吸収液は、流入口５からは第
１室３に至り、流出口５′からフローセル１の外
へ流出し、カラム７を経由して所定のイオン交換
反応を受けてのち吸収液排出口１０から排出され
る。また、カラム７の出口から吸収液排出口１０
へ至る流路の途中において、電極８，８′により
吸収液がイオン交換反応を受けて生成した物質の
導電率が測定される。更に、第２室４における被
測定液中の上記NH3ガスは、NH3ガス透過性膜
２を透過して第１室３に至つて上記吸収液と反応
する。吸収液がHCである場合について、第１室
３におけるNH3ガスと吸収液との反応を例示す
れば次式(3)のようになる
NH3＋HCl→NH4Cl ……(3)
上式(3)で生成したNH4Clは吸収液に運ばれて
カラム７に至り、該カラム７において吸収液（例
えばHCl）およびNH4Clが夫々下式(4)に示すよ
うなイオン交換反応を受ける。 The operation of the embodiment of the present invention will be explained below.
In the figure, the liquid to be measured introduced from the liquid to be measured inlet 11 passes through the immobilized enzyme 12, and urea in the liquid to be measured undergoes an enzymatic reaction as shown in the above formula (1) by the action of urease. After that, a reagent such as NaOH is added to the liquid to be measured from the reagent addition port 17 to make the liquid to be measured alkaline, and the NH 3 generated in the previous equation (1)
or NH 4 + changes to NH 3 gas. Thus, the liquid to be measured reaches the second chamber 4 from the inlet 6 and reaches the outlet 6'.
The liquid flows out of the flow cell 1 and is discharged to the outside from the liquid to be measured outlet 13. On the other hand, the absorption liquid introduced from the absorption liquid introduction port 14 reaches the first chamber 3 from the inflow port 5, flows out of the flow cell 1 from the outflow port 5', passes through the column 7, and performs a predetermined ion exchange process. After undergoing the reaction, it is discharged from the absorption liquid discharge port 10. Also, from the outlet of the column 7 to the absorption liquid outlet 10
On the way to the flow path, the electrodes 8 and 8' measure the conductivity of the substance produced by the ion exchange reaction of the absorption liquid. Further, the NH 3 gas in the liquid to be measured in the second chamber 4 passes through the NH 3 gas permeable membrane 2 to reach the first chamber 3 and reacts with the absorption liquid. When the absorption liquid is HC, an example of the reaction between the NH 3 gas and the absorption liquid in the first chamber 3 is as shown in the following equation (3): NH 3 +HCl→NH 4 Cl ...(3) Above equation The NH 4 Cl generated in (3) is carried by the absorption liquid and reaches column 7, where the absorption liquid (for example, HCl) and NH 4 Cl undergo an ion exchange reaction as shown in the following formula (4). receive.
（但し、は樹脂を示す記号である）上式(4)で生
成したH2OおよびNH4Clは、カラム７から導出
され吸収液排出口１０から排出されるが、カラム
７から吸収液排出口１０に至る間に上記電極８，
８′によつて夫々の導電率が検出される。すなわ
ち、上記吸収液中に上記第１室３でNH3が吸収
されないときは上記電極８，８′によつてH2Oの
導電率が連続的に検出されてベースラインの値と
なり、上記吸収液中に上記NH3が吸収されてく
ると上式(4)で生成したNH4CHが上記電極８，
８′によつてその導電率が検出されて検出値の値
となる。このようにして上記電極８，８′によつ
て検出された導電率に関する信号は、所定の信号
処理（図示せず）が施こされて被測定液中の尿素
濃度して表示器等（図示せず）に表示される。 (However, is a symbol indicating resin.) The H 2 O and NH 4 Cl generated in the above formula (4) are led out from the column 7 and discharged from the absorption liquid outlet 10; While reaching the outlet 10, the electrode 8,
8' detects the respective conductivity. That is, when NH 3 is not absorbed into the absorption liquid in the first chamber 3, the conductivity of H 2 O is continuously detected by the electrodes 8, 8' and becomes the baseline value, When the above NH 3 is absorbed into the liquid, the NH 4 CH generated in the above equation (4) is transferred to the above electrode 8,
The conductivity is detected by 8' and becomes a detected value. The signals related to conductivity detected by the electrodes 8 and 8' in this manner are subjected to predetermined signal processing (not shown) and converted to the urea concentration in the liquid to be measured on a display such as an indicator (see the figure). (not shown).
以上詳しく説明したような本発明の実施例によ
れば、カラム７内に充填された陰イオン交換樹脂
によつて吸収液等をイオン交換するような構成で
あるため、ベースラインがH2Oの導電率となつ
ており検出されるNH4OH導電率のＳ／Ｎ比が良
いという利点を有する。また、前記前出願におい
てはベースラインである吸収液の導電率が検出さ
れるNH4Clの導電率よりも大きいため、該ベー
スラインが周囲の温度変化に伴なつて変化し易い
という欠点があつたが、本発明実施例によれば吸
収液をイオン交換したのち導電率を検出している
ため、検出されるNH4OHの導電率の方がベース
ラインとなる物質の導電率よりも大きく、且つ周
囲の温度変化に伴なつてベースラインが変化する
ようなこともないという利点を有する。更に、本
発明の実施例によれば、前記従来例に比して、被
測定液中の尿素濃度を短時間で正確に測定できる
という利点も有している。 According to the embodiment of the present invention as described in detail above, since the structure is such that the absorption liquid etc. is ion-exchanged by the anion exchange resin packed in the column 7, the baseline is H 2 O. It has the advantage that the S/N ratio of the detected NH 4 OH conductivity is good. In addition, in the previous application, since the conductivity of the absorbing liquid, which is the baseline, is higher than the conductivity of the detected NH 4 Cl, there is a drawback that the baseline easily changes with changes in the surrounding temperature. However, according to the embodiment of the present invention, the conductivity is detected after ion-exchanging the absorption liquid, so the conductivity of the detected NH 4 OH is higher than the conductivity of the baseline substance. Another advantage is that the baseline does not change with changes in ambient temperature. Furthermore, the embodiment of the present invention has the advantage that the urea concentration in the liquid to be measured can be accurately measured in a short time compared to the conventional example.
図は本発明実施例の構成説明図である。
１……フローセル、２……NH3ガス透過性膜、
３，４……室、５，６……入口、５′，６′……出
口、７……カラム、８，８′……電極、１２……
固定化酵素、１５……容器、１６……吸収液。
The figure is a configuration explanatory diagram of an embodiment of the present invention. 1...Flow cell, 2... NH3 gas permeable membrane,
3, 4... Chamber, 5, 6... Inlet, 5', 6'... Outlet, 7... Column, 8, 8'... Electrode, 12...
Immobilized enzyme, 15... Container, 16... Absorption liquid.
Claims (1)
させる酵素反応によつてNH4 +を生成させ、該
NH4 +を前記被測定液へのアルカリ試薬添加によ
つてNH3に変換させ、その後該NH3を吸収液に
吸収させ、該吸収液の導電率変化量を検出して前
記尿素の濃度を間接的に測定する尿素測定方法に
おいて、前記吸収液を陰イオン交換樹脂へ透過せ
しめてのち導電率変化量を検出するようにしたこ
とを特徴とする尿素の測定方法。 ２ 吸収液が夫々導入・導出される導入口および
導出口を有しフローセル内の一側の流路を構成す
る第１室と、被測定液が夫々導入・導出される導
入口および導出口を有しフローセル内の他側の流
路を構成するとともに前記第１室とはNH3ガス
透過性膜を介して隣接する第２室と、被測定液中
の尿素に作用して酵素反応を生ぜしめるウレアー
ゼ酵素が固定化された固定化酵素と、該固定化酵
素から前記第２室の導入口に至る流路の途中に設
けられた試薬添加口と、前記第１室の導出口より
も下流に設けられ陰イオン交換樹脂が充填された
カラムと、該カラムの下流に設けられ該カラムを
透過してきた前記吸収液の導電率を検出する電極
とを具備し、前記吸収液の導電率変化量から間接
的に前記被測定液中の尿素濃度を測定する尿素測
定装置。[Claims] 1. NH 4 + is produced by an enzymatic reaction in which urease acts on urea contained in a liquid to be measured, and
NH 4 + is converted to NH 3 by adding an alkaline reagent to the liquid to be measured, and then the NH 3 is absorbed into an absorption liquid, and the change in conductivity of the absorption liquid is detected to determine the concentration of urea. A method for indirectly measuring urea, characterized in that the amount of change in conductivity is detected after the absorption liquid is permeated through an anion exchange resin. 2. A first chamber having an inlet and an outlet through which the absorption liquid is introduced and taken out, respectively, and forming a flow path on one side of the flow cell, and an inlet and an outlet through which the liquid to be measured is introduced and taken out, respectively. The second chamber is adjacent to the first chamber through an NH 3 gas permeable membrane, and acts on urea in the liquid to be measured to cause an enzyme reaction. an immobilized enzyme on which a urease enzyme is immobilized, a reagent addition port provided in the middle of a flow path from the immobilized enzyme to the inlet of the second chamber, and downstream of the outlet of the first chamber. a column provided in the column and filled with an anion exchange resin, and an electrode provided downstream of the column for detecting the conductivity of the absorption liquid that has passed through the column, and detecting the amount of change in the conductivity of the absorption liquid. A urea measuring device that indirectly measures the urea concentration in the liquid to be measured.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56163590A JPS5863853A (en) | 1981-10-14 | 1981-10-14 | Method and apparatus for measuring urea |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56163590A JPS5863853A (en) | 1981-10-14 | 1981-10-14 | Method and apparatus for measuring urea |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5863853A JPS5863853A (en) | 1983-04-15 |
| JPS6334429B2 true JPS6334429B2 (en) | 1988-07-11 |
Family
ID=15776804
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP56163590A Granted JPS5863853A (en) | 1981-10-14 | 1981-10-14 | Method and apparatus for measuring urea |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5863853A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4649671B2 (en) * | 2001-01-05 | 2011-03-16 | 学校法人早稲田大学 | Detection cell and enzyme sensing system in enzyme sensing |
-
1981
- 1981-10-14 JP JP56163590A patent/JPS5863853A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5863853A (en) | 1983-04-15 |
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