JPS6334427B2 - - Google Patents
Info
- Publication number
- JPS6334427B2 JPS6334427B2 JP56176112A JP17611281A JPS6334427B2 JP S6334427 B2 JPS6334427 B2 JP S6334427B2 JP 56176112 A JP56176112 A JP 56176112A JP 17611281 A JP17611281 A JP 17611281A JP S6334427 B2 JPS6334427 B2 JP S6334427B2
- Authority
- JP
- Japan
- Prior art keywords
- current
- electrophoresis
- sample
- electricity
- leading liquid
- 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
Links
- 238000001962 electrophoresis Methods 0.000 claims description 28
- 239000007788 liquid Substances 0.000 claims description 23
- 238000001514 detection method Methods 0.000 claims description 15
- 230000005611 electricity Effects 0.000 claims description 15
- 230000005012 migration Effects 0.000 claims description 15
- 238000013508 migration Methods 0.000 claims description 15
- 239000000203 mixture Substances 0.000 claims description 9
- 238000002347 injection Methods 0.000 claims description 5
- 239000007924 injection Substances 0.000 claims description 5
- 150000002500 ions Chemical class 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- 230000010354 integration Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 3
- 238000001649 capillary isotachophoresis Methods 0.000 description 2
- 238000002218 isotachophoresis Methods 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44717—Arrangements for investigating the separated zones, e.g. localising zones
- G01N27/4473—Arrangements for investigating the separated zones, e.g. localising zones by electric means
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44743—Introducing samples
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Physics & Mathematics (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】
この発明は電気泳動装置に関し、さらに詳しく
は、泳動路の任意位置を試料のイオン成分ゾーン
の前端が通過していることを検知しうる手段を備
えた電気泳動装置に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an electrophoresis apparatus, and more particularly, to an electrophoresis apparatus equipped with means capable of detecting that the front end of an ionic component zone of a sample is passing through an arbitrary position of an electrophoresis path. .
電気泳動装置において泳動路の所定の位置を試
料のイオン成分ゾーンの前端が通過していること
を検知する必要性は、特に試料から目的成分を分
取するとき、あるいは逆に目的外の成分を除去す
るときなどに生じる。従来、この検知手段として
は、特許庁文献No.76−53349号（Journal of
Chromatography.119、1976）の第10頁に記載の
ように電気泳動分析装置が本来備えている検出器
に前記検知手段の機能を兼ねさせたもの、あるい
は特公昭56−8302号に記載のように泳動路の所定
の位置に本来の検出器とは別個に同様の検出器を
設けたものが知られている。 In an electrophoresis device, it is necessary to detect that the front end of the ionic component zone of the sample passes through a predetermined position in the migration path, especially when separating the target component from the sample, or conversely, when separating undesired components from the sample. Occurs when removing. Conventionally, this detection means has been disclosed in Patent Office Document No. 76-53349 (Journal of
Chromatography.119, 1976), a detector originally equipped with an electrophoresis analyzer that also functions as the detection means, or as described in Japanese Patent Publication No. 56-8302. It is known that a similar detector is provided separately from the original detector at a predetermined position in the migration path.
しかしながら、前者の場合には、検出器が本来
設置される位置での検知しかできないので用途が
限定されており、後者の場合には、泳動路中にも
うひとつの検出器を設置せねばならないので装置
が複雑になる。 However, in the former case, the detection is limited to the position where the detector is originally installed, so its use is limited, and in the latter case, another detector must be installed in the migration path. The device becomes complicated.
さらに、検出器の出力信号は試料の組成や量に
よつて変動するので、その出力信号の意味を判別
する条件を試料ごとに選定してやる必要があり、
そうしなければ正確な検知を行えないことがあ
る。 Furthermore, since the output signal of the detector varies depending on the composition and amount of the sample, it is necessary to select conditions for determining the meaning of the output signal for each sample.
Otherwise, accurate detection may not be possible.
この発明は、このような状況に鑑みてなされた
もので、上記のような問題点を解消した電気泳動
装置を提供するものである。 The present invention has been made in view of the above circumstances, and aims to provide an electrophoresis device that solves the above-mentioned problems.
すなわち、この発明は、高電圧電源回路の両端
にそれぞれ接続されたターミナル液電極槽とリー
デイング液電極槽の間に連結した泳動路にて試料
を電気泳動させる電気泳動装置において、泳動電
流を検出する電流検出手段および検出された電流
を時間的に積算する電流積算手段を設けてなる電
気泳動装置を提供する。 That is, the present invention detects electrophoresis current in an electrophoresis apparatus in which a sample is electrophoresed in a migration path connected between a terminal liquid electrode tank and a leading liquid electrode tank connected to both ends of a high voltage power supply circuit. An electrophoresis apparatus is provided which includes a current detection means and a current integration means for temporally integrating the detected current.
上記電流検出手段は、具体的には例えば高電圧
電源回路の一端と電極槽の一つとを接続する接続
線に小抵抗を介設することで得られ、また上記電
流積算手段は前記小抵抗の両端電圧を積分する積
分器を設けることで得られる。 Specifically, the current detecting means can be obtained by, for example, inserting a small resistance in the connection line connecting one end of the high voltage power supply circuit and one of the electrode tanks, and the current integrating means can be obtained by interposing a small resistance in the connecting line connecting one end of the high voltage power supply circuit and one of the electrode tanks. This can be obtained by providing an integrator that integrates the voltage across both ends.
公知のように、電気泳動において供給された電
気量と一つのイオン成分ゾーンの泳動距離とは正
比例するものである。そこで電気量を知ることに
より泳動距離を知ることができる。今、リーデイ
ング液のイオン成分ゾーンに着目すれば、その泳
動距離と電気量の関係は、リーデイング液の組成
のみによつてきまる。ところがリーデイング液の
イオン成分ゾーンの後端は試料のイオン成分ゾー
ンの前端に他ならないから、試料のイオン成分ゾ
ーンの前端の泳動距離もリーデイング液のイオン
成分ゾーンの泳動距離と電気量の関係によつて知
ることができる。つまり、リーデイング液が同一
であるかぎり、同一の判断条件を用いて、電気量
から試料のイオン成分ゾーンの泳動距離を知るこ
とができ、試料の組成や量を考慮しなくてもよい
ことになる。 As is well known, the amount of electricity supplied during electrophoresis is directly proportional to the migration distance of one ionic component zone. Therefore, by knowing the amount of electricity, the migration distance can be determined. Now, if we focus on the ionic component zone of the leading liquid, the relationship between the migration distance and the amount of electricity depends only on the composition of the leading liquid. However, since the rear end of the ionic component zone of the leading liquid is nothing but the front end of the ionic component zone of the sample, the migration distance of the front end of the ionic component zone of the sample also depends on the relationship between the migration distance of the ionic component zone of the leading solution and the amount of electricity. You can know it. In other words, as long as the leading liquid is the same, the migration distance of the ionic component zone of the sample can be determined from the electrical quantity using the same judgment conditions, and there is no need to consider the composition or amount of the sample. .
前記電流積算手段の出力はまさに上記電気量に
他ならないから、結局、この発明の電気泳動装置
によれば、電流積算手段の出力によつて試料のイ
オン成分ゾーンの前端がどの位置まで泳動したか
を知ることができることとなる。しかも、検知位
置は任意であり、また出力信号の判別条件を試料
によつて変える必要もない。 Since the output of the current integration means is exactly the above-mentioned quantity of electricity, in the end, according to the electrophoresis apparatus of the present invention, it is possible to determine to what position the front end of the ionic component zone of the sample has migrated based on the output of the current integration means. You will be able to know. Moreover, the detection position is arbitrary, and there is no need to change the output signal discrimination conditions depending on the sample.
ところで、細管式等速電気泳動分析において分
析時間の短縮と精度のよい検出という２つの目的
を同時に達成するために、分析の初期には泳動電
流を大きくとつてイオンの泳動速度を大にして時
間の短縮を図り、分析の後期には泳動電流を小さ
くしてジユール熱の発生をおさえて精度よく検出
するという方法がよく用いられる。この方法で重
要なことは、泳動電流を切換えるタイミングを適
切にすることである。 By the way, in order to simultaneously achieve the two objectives of shortening the analysis time and highly accurate detection in capillary isotachophoresis analysis, the electrophoresis current is increased at the beginning of the analysis to increase the migration speed of ions. In order to shorten the time, a method is often used in which the electrophoresis current is reduced in the latter stages of the analysis to suppress the generation of Joule heat and to achieve accurate detection. What is important in this method is appropriate timing for switching the electrophoresis current.
このために、従来装置では、泳動路の所定位置
に別個の検出器を設けて、それにより試料のイオ
ン成分ゾーンの通過時刻を検知することが行われ
ていたが、前記したように、検出器の出力信号は
試料の組成や量によつてそれぞれ異なるので、試
料のイオン成分ゾーンの通過を判別する条件を試
料ごとに適切に選定しなければならないという問
題があつた。 For this purpose, in conventional devices, a separate detector is installed at a predetermined position in the migration path to detect the time when the sample passes through the ion component zone. Since the output signal differs depending on the composition and amount of the sample, there is a problem in that the conditions for determining whether the sample passes through the ionic component zone must be appropriately selected for each sample.
この発明の電気泳動装置では、前記したよう
に、判別条件を試料ごとに変更する必要がない。 In the electrophoresis apparatus of the present invention, as described above, there is no need to change the discrimination conditions for each sample.
従つて、この発明の電気泳動装置を利用してそ
の電流積算手段の出力信号に基づき泳動電流を切
換えるようにすれば、試料の如何にかかわらず、
一定の判別条件だけで正確なタイミングで泳動電
流を切換を行いうるようになる。 Therefore, if the electrophoresis apparatus of the present invention is used to switch the electrophoresis current based on the output signal of the current integration means, regardless of the sample,
It becomes possible to switch the electrophoresis current at accurate timing only under certain discrimination conditions.
以下、図に示す一実施例に基づいてこの発明を
詳説するが、ここで示される装置は上記のごとく
泳動電流を切換えるように構成した細管式等速電
気泳動分析装置である。 Hereinafter, the present invention will be explained in detail based on an embodiment shown in the drawings, and the apparatus shown here is a capillary type isotachophoresis analyzer configured to switch the electrophoresis current as described above.
１は細管式等速電気泳動分析装置で、直流高電
圧電源回路２の両端にそれぞれターミナル液電極
槽３とリーデイング液電極槽４とが接続され、こ
れら両電極槽３，４の間に試料注入部５と検出器
６とが管路７にて連結されている。試料注入部５
と検出器６の間の管路７は、管路径の太いプレチ
ユーブ７ａと管路径の細いキヤピラリーチユーブ
７ｂが段状部７ｃを介して直列に連結された２段
チユーブである。直流高電圧電源回路２の出力
は、制御回路８を介してマイクロコンピユータ９
により制御される。直流高電圧電源回路２とター
ミナル液電極槽３の間の接続線に介設されている
１０は電流検出回路であり、その出力がマイクロ
コンピユータ９に入力されている。１１は操作卓
である。 1 is a capillary type isotachophoresis analyzer, in which a terminal liquid electrode tank 3 and a leading liquid electrode tank 4 are connected to both ends of a DC high voltage power supply circuit 2, and a sample is injected between these electrode tanks 3 and 4. The section 5 and the detector 6 are connected through a conduit 7. Sample injection part 5
The pipe line 7 between the detector 6 and the detector 6 is a two-stage tube in which a pretube 7a with a large pipe diameter and a capillary reach tube 7b with a small pipe diameter are connected in series via a stepped portion 7c. The output of the DC high voltage power supply circuit 2 is sent to the microcomputer 9 via the control circuit 8.
controlled by A current detection circuit 10 is interposed in the connection line between the DC high voltage power supply circuit 2 and the terminal liquid electrode tank 3, and its output is input to the microcomputer 9. 11 is an operation console.
さて、この装置１のように分析用泳動路が２段
チユーブである場合には、試料のイオン成分ゾー
ンがプレチユーブ７ａを泳動するときに大電流を
供給し、試料のイオン成分ゾーンがキヤピラリー
チユーブ７ｂを泳動するときに小電流を供給する
ようにすることが、分析時間の短縮と精度のよい
検出を行う上で好ましいことである。そこで、試
料のイオン成分ゾーンの前端が段状部７ｃに到達
した時刻を知り、その時刻に泳動電流を大電流か
ら小電流に切換えるようにするのがよい。 Now, when the analytical migration path is a two-stage tube as in this apparatus 1, a large current is supplied when the ion component zone of the sample migrates through the pretube 7a, and the ion component zone of the sample migrates through the capillary tube 7a. It is preferable to supply a small current when electrophoresing 7b in order to shorten analysis time and perform accurate detection. Therefore, it is preferable to know the time when the front end of the ion component zone of the sample reaches the stepped portion 7c, and to switch the electrophoresis current from a large current to a small current at that time.
このために、あらかじめこの装置１では、リー
デイング液の組成とそのリーデイング液を使用し
たときにリーデイング液のイオン成分ゾーンの後
端が試料注入部５から段状部７ｃまで泳動するの
に要する電気量とが対応づけられてマイクロコン
ピユータ９に記憶されている。 For this purpose, in this apparatus 1, the composition of the leading liquid and the amount of electricity required for the rear end of the ion component zone of the leading liquid to migrate from the sample injection part 5 to the stepped part 7c when the leading liquid is used are determined in advance. and are stored in the microcomputer 9 in association with each other.
分析に当つては、まず、操作卓１１を介してリ
ーデイング液の組成をマイクロコンピユータ９に
入力する。そうすると、マイクロコンピユータ９
はそれに対応した電気量Ｑをメモリーから読み出
して内部的に設定する。次に、通常の手順により
試料注入部５にターミナル液とリーデイング液の
境界面を作り、試料を注入し、操作卓１１からス
タート指令を入力する。この指令によりマイクロ
コンピユータ９は制御回路８を介して直流高電圧
電源回路２を制御し、比較的に大きな泳動電流i1
（たとえば200〜300μA）を供給する。泳動電流i1
は、電流検出回路１０でマイクロコンピユータ９
にフイードバツクされている。マイクロコンピユ
ータ９は、この泳動電流i1を時間的に積算すると
ともに、積算によつて得られる電気量ｑと前記電
気量Ｑとを比較し、一致したときに泳動電流i1を
比較的に小さな泳動電流i2（たとえば20〜100μA）
に切換える。電気量ｑが電気量Ｑに一致したとき
は、先述したようにリーデイング液のイオン成分
ゾーンの後端が段状部７ｃに来たときであり、す
なわち試料のイオン成分ゾーンの前端が段状部７
ｃに到達したときである。 For analysis, first, the composition of the reading liquid is input to the microcomputer 9 via the console 11. Then, the microcomputer 9
reads the corresponding quantity of electricity Q from the memory and sets it internally. Next, an interface between the terminal liquid and the leading liquid is created in the sample injection section 5 according to the usual procedure, a sample is injected, and a start command is input from the operation console 11. Based on this command, the microcomputer 9 controls the DC high voltage power supply circuit 2 via the control circuit 8, and the relatively large electrophoretic current i 1
(e.g. 200-300μA). electrophoretic current i 1
is the current detection circuit 10 and the microcomputer 9
Feedback is provided to The microcomputer 9 integrates the electrophoretic current i 1 over time, compares the quantity of electricity q obtained by the integration with the quantity of electricity Q, and when they match, sets the electrophoretic current i 1 to a relatively small value. Electrophoretic current i 2 (e.g. 20-100 μA)
Switch to When the quantity of electricity q matches the quantity of electricity Q, it is when the rear end of the ionic component zone of the leading liquid has reached the stepped portion 7c as described above, that is, the front end of the ionic component zone of the sample has reached the stepped portion. 7
This is when it reaches c.
従つて、試料はまず大きな泳動電流i1によつて
プレチユーブ７ａ内を泳動し、次に小さな泳動電
流i2によつてキヤピラリーチユーブ７ｂ内を泳動
することになる。しかも、これは試料の組成や量
にかかわりなく正確に行われる。 Therefore, the sample first migrates within the pretube 7a due to the large electrophoretic current i1 , and then migrates within the capillary tube 7b due to the small electrophoretic current i2 . Moreover, this is done accurately regardless of the composition or amount of the sample.
以上の説明から理解されるように、この発明の
電気泳動装置によれば、試料のイオン成分ゾーン
の前端が泳動路の任意のある位置に到達したこと
を、試料の組成や量を考慮することなく正確に検
知することができる。そこで未知試料の分析に極
めて有用である。また、泳動電流の積算値のみを
問題にし瞬間値は問題にしないから、泳動電流が
定電流であれ刻々変化する電流であれ、いずれの
場合でも正確に上記検知を行いうる利点もある。 As can be understood from the above description, according to the electrophoresis apparatus of the present invention, it is possible to determine when the front end of the ion component zone of the sample has reached an arbitrary position in the migration path, taking into account the composition and amount of the sample. It can be detected accurately without any problems. Therefore, it is extremely useful for analyzing unknown samples. Further, since only the integrated value of the electrophoretic current is considered and the instantaneous value is not a problem, there is an advantage that the above-mentioned detection can be performed accurately regardless of whether the electrophoretic current is a constant current or a current that changes every moment.
第１図はこの発明の電気泳動装置の一実施例の
構成説明図である。
１……細管式等速電気泳動分析装置、２……直
流高電圧電源回路、３……ターミナル液電極槽、
４……リーデイング液電極槽、７……管路、８…
…制御回路、９……マイクロコンピユータ、１０
……電流検出回路、１１……操作卓。
FIG. 1 is an explanatory diagram of the configuration of an embodiment of the electrophoresis apparatus of the present invention. 1... Capillary isotachophoresis analyzer, 2... DC high voltage power supply circuit, 3... Terminal liquid electrode tank,
4... Leading liquid electrode tank, 7... Conduit, 8...
...Control circuit, 9...Microcomputer, 10
...Current detection circuit, 11...Operation console.
Claims (1)
ターミナル液電極槽とリーデイング液電極槽の間
に連結した試料注入部を有する泳動路にて試料を
電気泳動させる電気泳動装置において、 泳動路が管路径の太いプレチユーブと管路径の
細いキヤピラリーチユーブとが段状部を介して直
列に連結された２段チユーブで構成され、リーデ
イング液の組成とそのリーデイング液を使用した
ときにリーデイング液のイオン成分ゾーンの後端
が試料注入部から段状部まで泳動するのに要する
電気量とを対応させて記憶する記憶手段と、泳動
電流を検出する電流検出手段と、検出された電流
を時間的に積算する電流積算手段と、電流積算手
段から出力される電気量と記憶手段に記憶された
電気量とを比較し一致した場合に信号を出力する
比較手段と、比較手段の出力する信号により泳動
電流を切り換える電流切換手段とを具備したこと
を特徴とする電気泳動装置。[Scope of Claims] 1. In an electrophoresis device for electrophoresing a sample in an electrophoresis path having a sample injection section connected between a terminal liquid electrode tank and a leading liquid electrode tank connected to both ends of a high voltage power supply circuit, respectively. The electrophoresis path consists of a two-stage tube in which a pre-tube with a large pipe diameter and a capillary reach tube with a small pipe diameter are connected in series through a stepped part, and the composition of the leading liquid and when the leading liquid is used. a storage means for storing the amount of electricity required for the rear end of the ion component zone of the leading liquid to migrate from the sample injection part to the stepped part in correspondence with the amount of electricity; a current detection means for detecting the migration current; and a current detection means for detecting the electrophoresis current; a current integrating means for temporally integrating the electrical quantity; a comparing means for comparing the quantity of electricity output from the current integrating means with the quantity of electricity stored in the storage means and outputting a signal when they match; 1. An electrophoresis device comprising: current switching means for switching electrophoresis current according to a signal.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56176112A JPS5876751A (en) | 1981-10-31 | 1981-10-31 | electrophoresis device |
| US06/361,738 US4459198A (en) | 1981-07-27 | 1982-03-23 | Electrophoretic apparatus |
| DE8282102577T DE3270957D1 (en) | 1981-07-27 | 1982-03-26 | Electrophoretic apparatus |
| EP82102577A EP0070963B1 (en) | 1981-07-27 | 1982-03-26 | Electrophoretic apparatus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56176112A JPS5876751A (en) | 1981-10-31 | 1981-10-31 | electrophoresis device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5876751A JPS5876751A (en) | 1983-05-09 |
| JPS6334427B2 true JPS6334427B2 (en) | 1988-07-11 |
Family
ID=16007892
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP56176112A Granted JPS5876751A (en) | 1981-07-27 | 1981-10-31 | electrophoresis device |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5876751A (en) |
-
1981
- 1981-10-31 JP JP56176112A patent/JPS5876751A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5876751A (en) | 1983-05-09 |
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