JPH0740028B2 - Additive for blood glucose measurement - Google Patents
Additive for blood glucose measurementInfo
- Publication number
- JPH0740028B2 JPH0740028B2 JP62006967A JP696787A JPH0740028B2 JP H0740028 B2 JPH0740028 B2 JP H0740028B2 JP 62006967 A JP62006967 A JP 62006967A JP 696787 A JP696787 A JP 696787A JP H0740028 B2 JPH0740028 B2 JP H0740028B2
- Authority
- JP
- Japan
- Prior art keywords
- additive
- blood glucose
- blood
- monohalogenated
- buffer
- 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 - Lifetime
Links
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- Investigating Or Analysing Biological Materials (AREA)
Description
【発明の詳細な説明】 （産業上の利用分野） 本発明は，解糖を阻止することにより血糖を長時間にわ
たり初期濃度に維持することが可能で，かつ溶血作用の
ない血糖測定用添加剤に関する。DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Use) The present invention is an additive for blood glucose measurement, which can maintain the initial concentration of blood glucose for a long time by inhibiting glycolysis and has no hemolytic action. Regarding
（従来の技術） 各種臨床検査や病理学的研究のため血液成分，例えばグ
ルコース（血糖），グルコースの分解産物であるピルビ
ン酸の乳酸，の測定がなされている。特に血糖は，生体
のエネルギー源として最も重要な物質のひとつであり，
その測定方法は臨床化学分野における基本的測定手法の
ひとつとなっている。血糖測定時において，採取した全
血をそのまま放置すると，赤血球の解糖作用により血糖
値が低下するため，通常，解糖阻止剤が添加される。解
糖阻止剤としては，モノハロゲン化酢酸金属塩やフッ化
ナトリウム（NaF）が用いられる。しかしモノハロゲン
化酢酸金属塩は，それ自身，比較的不安定であるため，
保存中に分解して解糖阻止能が低下する。さらに，モノ
ハロゲン化酢酸金属塩が分解すると溶血作用を示すこと
が認められている。フッ化ナトリウムは，化合物自体が
溶血作用を有する。(Prior Art) Blood components such as glucose (blood sugar) and lactic acid of pyruvic acid, which is a decomposition product of glucose, are measured for various clinical tests and pathological studies. In particular, blood glucose is one of the most important substances as the energy source of the living body,
The measurement method is one of the basic measurement methods in the field of clinical chemistry. If the whole blood collected is left as it is at the time of measuring blood glucose, the glycolytic action of red blood cells lowers the blood glucose level, and therefore a glycolytic inhibitor is usually added. Monohalogenated acetic acid metal salts and sodium fluoride (NaF) are used as glycolytic inhibitors. However, monohalogenated metal acetate is relatively unstable in itself,
It decomposes during storage and its ability to inhibit glycolysis is reduced. Furthermore, it has been recognized that when the monohalogenated metal acetate is decomposed, it exhibits a hemolytic action. The compound itself of sodium fluoride has a hemolytic action.
（発明が解決しようとする問題点） 本発明は上記従来の欠点を解決するものでありその目的
とするところは，解糖阻止効果を有し、かつ経時的にそ
の解糖阻止能が低下することがなく，そして溶血作用の
ない血糖測定用の添加剤を提供することにある。(Problems to be Solved by the Invention) The present invention solves the above-mentioned conventional drawbacks, and an object of the present invention is to have a glycolytic inhibition effect and to decrease its glycolytic inhibition ability over time. It is to provide an additive for blood glucose measurement which has no hemolytic effect.
（問題点を解決するための手段および作用） 本発明の血糖測定用添加剤は，モノハロゲン化酢酸金属
塩およびpH緩衝剤を含有し，そのことにより上記目的が
達成される。(Means and Actions for Solving Problems) The blood glucose measuring additive of the present invention contains a monohalogenated metal acetate and a pH buffering agent, whereby the above object is achieved.
本発明の血糖測定用添加剤に含有されるモノハロゲン化
酢酸金属塩としては，モノヨード酢酸ナトリウム，モノ
クロロ酢酸カリウム，モノブロモ酢酸ナトリウムなどが
挙げられる。このモノハロゲン化酢酸金属塩は,SH基を
含む化合物の該SH基の部分をアルキル化することが知ら
れている。つまり，モノハロゲン化酢酸金属塩は，解糖
系においてSH基を有する酵素〔グリセルアルデヒド３−
リン酸デヒドロゲナーゼ（glyceraldehyde3−phosphate
dehydrogenase;G3PDH）および乳酸デヒドロゲナーゼ
（lactate dehydrogenase）〕をアルキル化することに
より失活させ，その結果，解糖阻止効果が得られる。こ
のモノハロゲン化酢酸金属塩とSH基との反応には，該モ
ノハロゲン化酢酸金属塩に由来するカルボキシルアニオ
ン（XCH2COO-）とSH基を有する酵素との静電相互作用が
大きな影響を与える。さらにこの反応速度は反応系のpH
により異なり,pHが高い程その反応速度が大きい。しか
し，モノハロゲン化酢酸金属塩は特にアルカリ側で不安
定である。モノハロゲン化酢酸金属塩自体の水溶液のpH
は７〜８であるが,pH7を越える状態で保存されると，該
化合物自身が分解のため反応速度は急激に低下する。そ
のため，本発明においては，モノハロゲン化酢酸金属塩
とともにpH緩衝剤が含有され，解糖阻止剤は水溶液の状
態であるとき，その全体のpHが３〜７となるように調整
される。本添加剤のpHは，好ましくは生理的条件により
近い６〜７である。このようなpH緩衝剤としては，リン
酸緩衝液，クエン酸緩衝液，クエン酸−リン酸二ナトリ
ウム緩衝液，クエン酸−クエン酸ナトリウム緩衝液，フ
タル酸水素カリウム−水酸化ナトリウム緩衝液，コハク
酸−水酸化ナトリウム緩衝液，塩酸緩衝液など；および
水を加えるとpH3〜７の緩衝液を形成しうる化合物が挙
げられる。上記モノハロゲン化酢酸金属塩と緩衝剤との
配合比は特に限定されず，モノハロゲン化酢酸金属塩が
水溶液中においてpH3〜７の状態で存在しうるような量
の緩衝剤が存在すればよい。Examples of the monohalogenated acetic acid metal salt contained in the blood glucose measurement additive of the present invention include sodium monoiodoacetate, potassium monochloroacetate, sodium monobromoacetate and the like. This monohalogenated metal acetate is known to alkylate a portion of the SH group of a compound containing the SH group. In other words, the monohalogenated acetic acid metal salt is an enzyme having an SH group in the glycolysis system [glyceraldehyde 3-
Phosphate dehydrogenase (glyceraldehyde3-phosphate
dehydrogenase (G3PDH) and lactate dehydrogenase (lactate dehydrogenase)] are deactivated by alkylation, resulting in a glycolytic inhibition effect. The reaction of the monohalogenated metal acetate and SH groups, carboxyl anions derived from the monohalogenated acetic acid metal salt (XCH 2 COO -) electrostatic interaction with the enzyme having a SH group a significant impact give. Furthermore, this reaction rate depends on the pH of the reaction system.
The higher the pH, the higher the reaction rate. However, monohalogenated acetic acid metal salts are unstable especially on the alkaline side. PH of aqueous solution of monohalogenated metal acetate
Is 7 to 8, but when stored in a state of exceeding pH 7, the reaction rate rapidly decreases due to decomposition of the compound itself. Therefore, in the present invention, a pH buffer is contained together with the monohalogenated acetic acid metal salt, and the glycolysis inhibitor is adjusted so that the total pH thereof is 3 to 7 when in an aqueous solution state. The pH of this additive is preferably 6 to 7, which is closer to physiological conditions. Such pH buffers include phosphate buffer, citrate buffer, citric acid-disodium phosphate buffer, citric acid-sodium citrate buffer, potassium hydrogen phthalate-sodium hydroxide buffer, amber. Acid-sodium hydroxide buffer, hydrochloric acid buffer and the like; and compounds capable of forming a buffer having a pH of 3 to 7 when water is added. The compounding ratio of the metal monohalogenated acetic acid salt and the buffer is not particularly limited as long as the amount of the buffered metal monohalogenated acetic acid metal salt can be present in an aqueous solution at a pH of 3 to 7. .
本発明の解糖阻止剤は,EDTAやヘパリンのような抗凝固
剤と併用することも可能である。本添加剤は，モノハロ
ゲン化酢酸金属塩と上記緩衝剤とを混合した粉末状の形
態であってもよく，水溶液の状態であってもよい。この
添加剤は，血液に添加して用いられる他，あらかじめ常
圧もしくは真空採血管の内部に収容もしくは管壁内面に
付与しておくこともできる。このような採血管には，さ
らに血清分離剤などが同時に収容されていてもよい。本
発明の添加剤においては，モノハロゲン化酢酸金属塩が
pH緩衝液により所定のpHに保たれるため，該モノハロゲ
ン化酢酸金属塩が保存中に分解することが極めて少な
い。そのため，血液と混合されたときに，解糖阻止能が
低下することがない。さらに，モノハロゲン化酢酸金属
塩の分解による溶血作用がない。The glycolytic inhibitor of the present invention can be used in combination with an anticoagulant such as EDTA or heparin. The present additive may be in the form of a powder in which a monohalogenated acetic acid metal salt is mixed with the above buffer, or may be in the form of an aqueous solution. In addition to being used by adding it to blood, this additive can be housed in a normal pressure or vacuum blood collection tube or applied to the inner surface of the tube wall in advance. Such a blood collection tube may further contain a serum separating agent or the like at the same time. In the additive of the present invention, the monohalogenated acetic acid metal salt is
Since the pH is kept at a predetermined level by the pH buffer solution, the monohalogenated metal acetate is hardly decomposed during storage. Therefore, when it is mixed with blood, the ability to inhibit glycolysis does not decrease. In addition, there is no hemolytic action due to the decomposition of monohalogenated metal acetate.
本発明の添加剤は，血液1mlあたりモノハロゲン化酢酸
金属塩が0.1〜0.6mgの割合となるように血液と混合され
る。本発明の添加剤が加えられた血液は，例えば22〜23
℃において保存した場合,20〜24時間にわたり安定した
値で血糖の測定が可能である。保存された血液は通常，
遠心分離により血球を除去して血漿を得，ブドウ糖酸化
酵素法などにより血糖が測定される。The additive of the present invention is mixed with blood at a ratio of 0.1-0.6 mg of metal monohalogenated acetic acid salt per 1 ml of blood. Blood to which the additive of the present invention is added is, for example, 22 to 23.
When stored at ℃, blood glucose can be measured at a stable value for 20 to 24 hours. The stored blood is usually
Blood cells are removed by centrifugation to obtain plasma, and blood glucose is measured by the glucose oxidase method or the like.
（実施例） 以下に本発明を実施例につき説明する。(Example) Hereinafter, the present invention will be described with reference to Examples.
実施例 リン酸緩衝液5mlにモノヨード酢酸ナトリウム0.25gおよ
びヘパリンナトリウム2000ユニットを加えて溶解させ
た。この溶液のpHは6.55であった。これを22〜23℃にて
１ヶ月間保存した後,50μを採取して市販の2ml用採血
管に収容した。これにヒト新鮮血2mlを採取し，転倒混
和し,3000rpmで５分間遠心分離して血漿を得，溶血の有
無を観察した。この血漿の血糖値を臨床検査法提要に記
載された電極法によるブドウ糖測定法を用いて測定し
た。同様の方法でヒト新鮮血を採取し転倒混和後,22〜2
3℃にて12時間放置した後に遠心分離して溶血の有無を
観察後，血糖値の測定を行った。さらに24時間放置した
ものについても同様に測定を行った。その結果を表１お
よび表２に示す。Example To 5 ml of a phosphate buffer, 0.25 g of sodium monoiodoacetate and 2000 units of sodium heparin were added and dissolved. The pH of this solution was 6.55. This was stored at 22 to 23 ° C. for 1 month, and 50 μm was collected and placed in a commercially available 2 ml blood collection tube. 2 ml of human fresh blood was collected, mixed by inversion, centrifuged at 3000 rpm for 5 minutes to obtain plasma, and the presence or absence of hemolysis was observed. The blood glucose level of this plasma was measured using the glucose measurement method by the electrode method described in the clinical test method guidelines. Human fresh blood was collected by the same method and mixed by inverting, then
After standing at 3 ° C for 12 hours, centrifugation was performed to observe the presence or absence of hemolysis, and then the blood glucose level was measured. Further, the same measurement was performed for the one left for 24 hours. The results are shown in Tables 1 and 2.
比較例 リン酸緩衝液の代わりに蒸留水を用いたこと以外は実施
例と同様である。その結果を表１および表２に示す。Comparative Example The same as the example except that distilled water was used instead of the phosphate buffer. The results are shown in Tables 1 and 2.
（発明の効果） 本発明によれば，このように，解糖阻止能を有し，かつ
長時間の保存が可能であり，そして溶血を生じない血糖
測定用添加剤が得られる。この添加剤を使用すると，血
液採取後の放置時間にかかわらず血糖値が正確に測定さ
れる。そのため，各種臨床検査や病理学的研究のために
好適に利用され得る。 (Effect of the Invention) According to the present invention, as described above, an additive for blood glucose measurement, which has a glycolytic inhibition ability, can be stored for a long time, and does not cause hemolysis, can be obtained. When this additive is used, the blood glucose level can be accurately measured regardless of the standing time after blood collection. Therefore, it can be suitably used for various clinical tests and pathological studies.
Claims (2)
を含有する血糖測定用添加剤。1. An additive for measuring blood glucose, which comprises a monohalogenated metal acetate and a pH buffer.
１項に記載の添加剤。2. The additive according to claim 1, which has a pH in the range of 3 to 7.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62006967A JPH0740028B2 (en) | 1987-01-14 | 1987-01-14 | Additive for blood glucose measurement |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62006967A JPH0740028B2 (en) | 1987-01-14 | 1987-01-14 | Additive for blood glucose measurement |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63173966A JPS63173966A (en) | 1988-07-18 |
| JPH0740028B2 true JPH0740028B2 (en) | 1995-05-01 |
Family
ID=11652974
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62006967A Expired - Lifetime JPH0740028B2 (en) | 1987-01-14 | 1987-01-14 | Additive for blood glucose measurement |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0740028B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0203280D0 (en) * | 2002-02-12 | 2002-03-27 | Ic Innovations Ltd | Anti-glycolytic composition |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0687062B2 (en) * | 1985-05-10 | 1994-11-02 | 株式会社京都医科学研究所 | How to prevent glycolysis in blood |
-
1987
- 1987-01-14 JP JP62006967A patent/JPH0740028B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPS63173966A (en) | 1988-07-18 |
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