JPH0675066B2 - Catecholamine detector - Google Patents
Catecholamine detectorInfo
- Publication number
- JPH0675066B2 JPH0675066B2 JP59089139A JP8913984A JPH0675066B2 JP H0675066 B2 JPH0675066 B2 JP H0675066B2 JP 59089139 A JP59089139 A JP 59089139A JP 8913984 A JP8913984 A JP 8913984A JP H0675066 B2 JPH0675066 B2 JP H0675066B2
- Authority
- JP
- Japan
- Prior art keywords
- detector
- sample
- catecholamine
- electrochemical
- catecholamines
- 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 - Fee Related
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
【発明の詳細な説明】 イ．技術の利用分野 本発明は、高速液体クロマトグラフィーに適した検出装
置、より詳しくは微量のカテコールアミン類やその代謝
物を測定する検出装置に関する。Detailed Description of the Invention a. TECHNICAL FIELD The present invention relates to a detector suitable for high performance liquid chromatography, and more particularly to a detector for measuring a trace amount of catecholamines and metabolites thereof.
ロ．従来技術 ３・４−ジヒドロキシフェノール骨核を持つ生体アミン
等のフェノール核を含む化学物質は、生体活性を起す物
質であるため、生化学分野ではこの物質の分析が重大な
課題となっている。B. 2. Description of the Related Art Since a chemical substance containing a phenol nucleus such as a biogenic amine having a 3,4-dihydroxyphenol bone nucleus is a substance which causes bioactivity, analysis of this substance has become a serious problem in the biochemical field.
ところで、このフェノール核を含む化学物質の分析は、
電気化学的検出器、いわゆるEDCを使用したり、トリハ
イドロオキシインドール法が用いられている。前者の手
法では、フェノール核を含んだ物質に対する感度は高い
が、個々の成分に対する特異性が低いため物質の特定が
困難であり、また後者の手段では、アドレナリンやノル
アドレナリンに対する感度、及び特異性は高いが、ドー
パやドーパミンに対する感度が低いという一長一短があ
り、生体組織中に微量しか含まれないこの種の物質を定
性定量的に分析することが困難であるといった問題があ
った。By the way, the analysis of chemical substances containing this phenol nucleus
Electrochemical detectors, so-called EDC, are used, or the trihydrooxindole method is used. The former method has high sensitivity to substances containing phenolic nuclei, but it is difficult to identify the substance because of low specificity to individual components, and the latter method has low sensitivity and specificity to adrenaline and noradrenaline. Although it is high, it has disadvantages such as low sensitivity to dopa and dopamine, and there is a problem that it is difficult to qualitatively and quantitatively analyze this kind of substance that is contained in a living tissue in a trace amount.
ハ．目的 本発明はこのような問題に鑑み、カテコールアミン類、
及びその代謝物質を高い感度で、特異的に分析すること
ができる新規な検出装置を提供することにある。C. OBJECT In view of such problems, the present invention has catecholamines,
Another object of the present invention is to provide a novel detection device capable of specifically analyzing a metabolite thereof with high sensitivity.
ニ．発明の構成 すなわち、本発明の特徴とするところは、電気化学的検
出器の酸化、還元反応を利用してトリハイドロオキシイ
ンドール法を効果的に適用した点にある。D. Structure of the Invention That is, the feature of the present invention resides in that the trihydrooxyindole method is effectively applied by utilizing the oxidation and reduction reactions of the electrochemical detector.
ホ．実施例 そこで、以下に本発明の詳細を図示した実施例に基づい
て説明する。E. Examples Therefore, details of the present invention will be described below based on illustrated examples.
第１図は、本発明の実施例を示す装置の構成図であっ
て、図中符号１は、本発明の特徴部分をなす検出装置
で、試料流入口側から電気化学的検出器11、ミキシング
コイル12、蛍光検出器13がパイプにより直列に接続し、
またミキシングコイル12には分岐管により送液ポンプ２
を介してアルカリ試薬を収容した反応液槽３がパイプに
より接続している。４は、中央制御装置で、装置全体の
動作タイミングの制御、及び検出信号の処理を行なうも
のである。なお、図中符号５は、試料注入口６がポンプ
７を介してキャリア液槽８に、吐出口側が検出装置１に
接続する高速液体クロマトグラフィ用カラムを示してい
る。FIG. 1 is a block diagram of an apparatus showing an embodiment of the present invention. In the figure, reference numeral 1 is a detection device which is a characteristic part of the present invention. An electrochemical detector 11 and a mixing device are provided from the sample inlet side. The coil 12 and the fluorescence detector 13 are connected in series by a pipe,
In addition, the mixing coil 12 is provided with a liquid delivery pump 2 by a branch pipe
A reaction solution tank 3 containing an alkaline reagent is connected via a pipe. A central control unit 4 controls the operation timing of the entire device and processes detection signals. In the figure, reference numeral 5 indicates a high performance liquid chromatography column in which the sample injection port 6 is connected to the carrier liquid tank 8 via the pump 7 and the ejection port side is connected to the detection device 1.
この実施例において、試料注入口６から分析すべき試料
をカラム５に注入すると、成分に応じて分離され、検出
装置１に流入する。電気化学的検出器11は、成分に分離
され試料をその電気化学作用によって酸化−還元し、成
分に応じた電気信号を出力する。この酸化−還元された
成分は、ミキシングコイル12を経由して蛍光検出器13に
流入する。このようにして電気化学的検出器11からの出
力を基にしてスクリーニングを行ない、目的成分が検知
された時点で、ポンプ２を作動して反応液槽３からアル
カリ試薬をミキシングコイル12に注入すると、電気化学
的検出器11により予め酸化−還元を受けている目的成分
は、ミキシングコイル12を経由しながら反応液と十分に
攪拌されて蛍光反応を起して特異性を付与され蛍光検出
器13により検出される。In this embodiment, when the sample to be analyzed is injected from the sample injection port 6 into the column 5, it is separated according to the components and flows into the detection device 1. The electrochemical detector 11 oxidizes and reduces the sample separated into its components by its electrochemical action, and outputs an electric signal corresponding to the component. This oxidized-reduced component flows into the fluorescence detector 13 via the mixing coil 12. In this way, screening is performed based on the output from the electrochemical detector 11, and when the target component is detected, the pump 2 is operated to inject the alkaline reagent from the reaction liquid tank 3 into the mixing coil 12. , The target component that has been previously subjected to oxidation-reduction by the electrochemical detector 11 is sufficiently stirred with the reaction solution while passing through the mixing coil 12 to cause a fluorescent reaction to give specificity and the fluorescence detector 13 Detected by.
［実施例］ 強イオン交換樹脂を固定相に、リン酸二水素ナトリウム
（0.06M,PH4.3）を移動相とする高速液体クロマトグラ
フ用カラムにカテコールアミン類を含む試料を注入した
ところ、α−メチルドーパ、ドーパ、ノルアドレナリ
ン、アドレナリンドーパミンの順序で分離されて電気化
学的検出器に流入した。このように分離された試料は、
電気化学的検出器内において酸化−還元反応を受け、 となった。次いでミキシングコイルに流入した時点で４
〜６規定のNa・OHを反応液として0.2〜0.5ml/分の割合
で送給した。これにより、例えばノルアドレナリンに例
を採ると、 なる反応により蛍光体に変換され、波長420nm〜520nmの
紫外線の照射を受けて特異的に蛍光を発した。他の成分
も同様に特異的に蛍光を発生し、第２図にしたようにα
−メチルドーパ、ドーパ、ノルアドレナリン、ドーパミ
ンがそれぞれ独立した状態で検出することができた。[Example] A sample containing catecholamines was injected into a column for high performance liquid chromatography using a strong ion exchange resin as a stationary phase and sodium dihydrogen phosphate (0.06M, PH4.3) as a mobile phase. Methyldopa, dopa, noradrenaline, adrenaline and dopamine were separated in this order and flowed into the electrochemical detector. The sample separated in this way is
Undergoes an oxidation-reduction reaction in the electrochemical detector, Became. Then when it flows into the mixing coil, 4
~ 6 N NaOH was fed as a reaction solution at a rate of 0.2 to 0.5 ml / min. Thus, taking noradrenaline as an example, It was converted to a phosphor by the reaction, and when it was irradiated with ultraviolet rays having a wavelength of 420 nm to 520 nm, it specifically emitted fluorescence. Other components similarly generate fluorescence specifically, and as shown in FIG.
-Methyldopa, dopa, noradrenaline and dopamine could be detected independently.
ヘ．効果 以上、説明したように本発明によれば、高速液体クロマ
トグラフィにより分離されたカテコールアミン類を、電
気化学的検出器とトリハイドロオキシインドール法を併
用して検出するようにしたので、電気化学的検出器によ
りスクリーニングを行なうことができるばかりでなく、
試料の酸化−還元によりトリハイドロオキシインドール
法が効果的に機能して微量のカテコールアミン類やその
代謝物を特異的かつ定量的に検出することができる。F. Effect As described above, according to the present invention, catecholamines separated by high performance liquid chromatography are detected by using an electrochemical detector in combination with the trihydroxyindole method. Not only can screening be performed with a vessel,
Due to the oxidation-reduction of the sample, the trihydroxyindole method effectively functions and a trace amount of catecholamines and their metabolites can be detected specifically and quantitatively.
第１図は、本発明の一実施例を示す装置の構成図、第２
図は、同上装置による分析結果の一例を示す説明図であ
る。 11……電気化学的検出器 12……ミキシングコイル 13……蛍光検出器、３……反応液槽 ４……高速液体クロマト用カラムFIG. 1 is a block diagram of an apparatus showing an embodiment of the present invention, and FIG.
The figure is an explanatory view showing an example of an analysis result by the same apparatus. 11 …… Electrochemical detector 12 …… Mixing coil 13 …… Fluorescence detector 3 …… Reaction solution tank 4 …… High performance liquid chromatography column
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.5 識別記号 庁内整理番号 ＦＩ 技術表示箇所 Ｇ０１Ｎ 33/50 Ｂ 7055−2Ｊ ─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. 5 Identification code Office reference number FI technical display location G01N 33/50 B 7055-2J
Claims (1)
電気化学的検出手段、該手段から流出する試料成分にア
ルカリ試薬を混合する手段、及び蛍光検出手段をそれぞ
れ上流側から直列に接続してなるカテコールアミン類検
出装置。1. An electrochemical detection means for detecting a component based on an oxidation-reduction reaction, a means for mixing an alkaline reagent with a sample component flowing out from the means, and a fluorescence detection means are respectively connected in series from the upstream side. A catecholamines detector.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59089139A JPH0675066B2 (en) | 1984-05-02 | 1984-05-02 | Catecholamine detector |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59089139A JPH0675066B2 (en) | 1984-05-02 | 1984-05-02 | Catecholamine detector |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS60233548A JPS60233548A (en) | 1985-11-20 |
| JPH0675066B2 true JPH0675066B2 (en) | 1994-09-21 |
Family
ID=13962537
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59089139A Expired - Fee Related JPH0675066B2 (en) | 1984-05-02 | 1984-05-02 | Catecholamine detector |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0675066B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0781999B2 (en) * | 1986-03-27 | 1995-09-06 | 株式会社島津製作所 | High performance liquid chromatograph for analysis of catecholamines |
| JP2711976B2 (en) * | 1993-03-15 | 1998-02-10 | 日立ソフトウエアエンジニアリング株式会社 | Gel electrophoresis pattern reader |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS57191548A (en) * | 1981-05-20 | 1982-11-25 | Yanagimoto Seisakusho:Kk | Analysing apparatus of catechol amine |
| JPS5827062A (en) * | 1981-08-12 | 1983-02-17 | Sekisui Chem Co Ltd | Liquid chromatograph for measuring catecholamine |
-
1984
- 1984-05-02 JP JP59089139A patent/JPH0675066B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPS60233548A (en) | 1985-11-20 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |