JPH0535825B2 - - Google Patents
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- JPH0535825B2 JPH0535825B2 JP60035095A JP3509585A JPH0535825B2 JP H0535825 B2 JPH0535825 B2 JP H0535825B2 JP 60035095 A JP60035095 A JP 60035095A JP 3509585 A JP3509585 A JP 3509585A JP H0535825 B2 JPH0535825 B2 JP H0535825B2
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- 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
- G01N27/44747—Composition of gel or of carrier mixture
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- Electrochemistry (AREA)
- Physics & Mathematics (AREA)
- Dispersion Chemistry (AREA)
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- Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Description
【発明の詳細な説明】
(イ) 産業上の利用分野
この発明は、電気泳動分析方法に関する。さら
に詳しくは、微量のイオン化物質を含有する被検
液を等速電気泳動分析に付してイオン性物質の分
離分析を行なう際の改良された方法に関する。[Detailed Description of the Invention] (a) Field of Industrial Application This invention relates to an electrophoretic analysis method. More specifically, the present invention relates to an improved method for separating and analyzing ionic substances by subjecting a sample liquid containing a trace amount of ionized substances to isokinetic electrophoresis analysis.
(ロ) 従来技術
従来から、種々のイオン性物質を分離分析する
方法として、等速電気泳動分析が行なわれてい
る。そして、非水溶性の液状媒体中にイオン性物
質を含有する試料を等速電気泳動に付す場合、こ
の試料を、アセトン等の水と自由に混合しうる親
水性有機溶媒に予め混合溶解させることによりそ
の極性を向上させ、これを被検液として等速電気
泳動分析用の電解液中すなわちリーデイング液と
ターミナル液との界面付近に注入して泳動を行な
う方法が汎用されている。(b) Prior Art Conventionally, isotachophoresis analysis has been used as a method for separating and analyzing various ionic substances. When subjecting a sample containing an ionic substance in a non-aqueous liquid medium to isokinetic electrophoresis, the sample must be mixed and dissolved in advance in a hydrophilic organic solvent such as acetone that can be freely mixed with water. A commonly used method is to improve the polarity of the sample and inject it as a test liquid into an electrolytic solution for isotachophoresis analysis, that is, near the interface between a leading solution and a terminal solution, and perform electrophoresis.
しかしながら、この手法では、イオン性物質を
液状媒体中に微量含有する試料を対象とする場
合、被検液中のイオン性物質の量が少ないから通
電性が悪くそれにより発生するジユール熱によつ
て気泡が生成したり分析効率も低下するという問
題点があつた。また、上記電解液中で非水溶性の
液状媒体が分相や沈殿して分析精度の低下や泳動
管の閉塞を招く場合もあつた。 However, when using this method to target a sample that contains a small amount of ionic substances in a liquid medium, the small amount of ionic substances in the test liquid results in poor electrical conductivity and the resulting Joule heat. There were problems such as the generation of bubbles and a decrease in analysis efficiency. Furthermore, in some cases, the non-aqueous liquid medium in the electrolyte solution undergoes phase separation or precipitation, resulting in a decrease in analysis accuracy and clogging of the electrophoresis tube.
そのため、泳動前に上記被検液と泳動用の電解
液（通常、リーデイング液）とを混合することに
より、被検液中のイオン性物質の量を上昇させる
と共に、分相や沈殿する前記液状媒体を予め分離
した後、この電解液含有溶液を注入して泳動を行
なう方法が提案されている。 Therefore, by mixing the above-mentioned test solution and an electrolytic solution for electrophoresis (usually a leading solution) before electrophoresis, the amount of ionic substances in the test solution can be increased, and the liquid may undergo phase separation or precipitation. A method has been proposed in which the medium is separated in advance and then this electrolyte-containing solution is injected to perform electrophoresis.
しかしながら、上記電解液含有溶液を被検液と
して用いた場合、その中には分析を意図するイオ
ンと同極性で易動度の高いイオン性物質（通常、
リーデイングイオン）が存在しておりかつこれら
が先に泳動されるため、目的のイオン性物質の検
知時間が長く分析時間が長いという問題点があつ
た。この時間をできるだけ短縮化するためには加
える泳動用の電解液の量をできるだけ少なくする
必要があるが、そのためには被検液の注入量を少
なくする必要があつて結果的に試料導入量を多く
することができず感度が低下する不都合が生じ
た。 However, when the above-mentioned electrolyte-containing solution is used as a test solution, it contains ionic substances (usually
Since there are leading ions (leading ions) and these are migrated first, there is a problem that it takes a long time to detect the ionic substance of interest and the analysis time is long. In order to shorten this time as much as possible, it is necessary to reduce the amount of electrolyte solution added for electrophoresis as much as possible, but to do so, it is necessary to reduce the amount of sample solution to be injected, and as a result, the amount of sample introduced must be reduced. This resulted in the inconvenience of lowering the sensitivity because it was not possible to increase the amount.
一方、非水溶性の液状媒体のみならず水溶性の
液状媒体ことに水溶性溶媒中に微量のイオン性物
質を含有する試料や被検液についても、通電性の
点で前述と同様な問題点があつた。 On the other hand, not only water-insoluble liquid media, but also water-soluble liquid media, and samples and test solutions containing trace amounts of ionic substances in water-soluble solvents have the same problems in terms of electrical conductivity. It was hot.
(ハ) 発明の目的
この発明の上記のごとき問題点を解消すべくな
されたものであり、溶媒中に微量のイオン性物質
を含有してなる被検液を等速電気泳動分析に付す
際に、通電性によるトラブルを生じることなくし
かも高感度でかつ簡便に所望のイオン性物質群を
分析できる電気泳動分析方法を提供しようとする
ものである。(c) Purpose of the Invention This invention has been made to solve the above-mentioned problems of the present invention, and is intended to solve the above-mentioned problems of the present invention. The object of the present invention is to provide an electrophoretic analysis method that can easily analyze a desired group of ionic substances with high sensitivity and without causing troubles due to conductivity.
(ニ) 発明の構成
かくしてこの発明によれば、溶媒中に微量のイ
オン性物質を含有してなる被検液を等速電気泳動
分析に付して該溶媒中の所望のイオン性物質を分
析するに際し、上記被検液中に、分析を意図する
イオンと逆性のイオンとなりかつ上記溶媒及び水
に良溶性のイオン性化合物を加えて等速電気泳動
を行なうことを特徴とする電気泳動分析方法が提
供される。(d) Structure of the Invention According to the present invention, a desired ionic substance in the solvent is analyzed by subjecting a test liquid containing a trace amount of an ionic substance in a solvent to isokinetic electrophoresis analysis. An electrophoretic analysis characterized in that when performing isokinetic electrophoresis, an ionic compound is added to the test solution that forms an ion with the opposite polarity to the ion intended for analysis and is well soluble in the solvent and water. A method is provided.
この発明の最も特徴とする点は、被検液中に分
析対象イオンと逆性の特定のイオン性化合物のみ
を実質的に加えておくことにより、分析対象イオ
ンと同極性及び逆性のイオン性物質を含有する泳
動用の電解液ことにリーデイング液を加えること
なく通電性を向上させ、それにより試料の導入量
の増加による感度向上、気泡発生の防止及び分析
時間の短縮化を可能とした点にある。 The most distinctive feature of this invention is that by substantially adding only a specific ionic compound with the opposite polarity to the analyte ion to the sample solution, it is possible to Improved conductivity without adding a leading solution to the electrophoresis solution containing the substance, thereby improving sensitivity by increasing the amount of sample introduced, preventing bubble generation, and shortening analysis time. It is in.
この発明において被検液中に共存させる逆性の
イオン性化合物は、被検液を構成する媒体すなわ
ち微量のイオン性物質に対する溶媒に対して溶解
度が高く、かつ水に対しても高溶解性の化合物で
あることを要する。さらにかかるイオン性化合物
としては分析を意図するイオンの逆性のイオンと
なるものが選択される。ここで「逆性のイオンと
なる」とはそれ自体が水性媒体中で解離してアニ
オン又はカチオンとなりうるものでありかつ水素
イオン及び水酸基イオン以外の対イオンを実質的
に生じないことを意味する。これらの逆性イオン
は易動度40〜80×10-5cmV-1sec-1のものが好まし
い。これらのイオン性化合物としては、例えば、
アニオンを分析対象とする場合、エチレンジアミ
ン、ジエチレントリアミン、トリエチレンテトラ
ミン、モノエタノールアミン、ジエタノールアミ
ン、トリエタノールアミン、イミダゾール等のア
ミン類が挙げられ、カチオンを分析対象とする場
合、塩酸、硝酸等の無機酸やシユウ酸、リンゴ
酸、乳酸、蟻酸、酢酸等の有機酸が挙げられる。 In this invention, the opposite ionic compound coexisting in the test solution has high solubility in the medium constituting the test solution, that is, the solvent for trace amounts of ionic substances, and has high solubility in water. Must be a compound. Furthermore, such an ionic compound is selected to have an ion having the opposite polarity to the ion intended for analysis. Here, "becoming a reverse ion" means that it can dissociate itself into an anion or cation in an aqueous medium, and that it does not substantially generate counter ions other than hydrogen ions and hydroxyl ions. . These reverse ions preferably have a mobility of 40 to 80×10 −5 cmV −1 sec −1 . These ionic compounds include, for example,
When anions are to be analyzed, amines such as ethylenediamine, diethylenetriamine, triethylenetetramine, monoethanolamine, diethanolamine, triethanolamine, and imidazole are included; when cations are to be analyzed, inorganic acids such as hydrochloric acid and nitric acid are included. Examples include organic acids such as oxalic acid, malic acid, lactic acid, formic acid, and acetic acid.
上記イオン性化合物の被検液中への導入量は、
該被検液中の微量イオン性物質の量によつても異
なるが通常、１〜20meq／とするのが適してお
り５〜10meq／とするのが好ましい。1meq／
未満の場合は通電性の点で適さず、50meq／
を超える場合は泳動速度の点で不適当である。 The amount of the above ionic compound introduced into the test solution is
Although it varies depending on the amount of trace ionic substances in the test liquid, it is usually suitable to be 1 to 20 meq/, and preferably 5 to 10 meq/. 1meq/
If it is less than 50meq/
If it exceeds , it is inappropriate in terms of migration speed.
一方、この発明の対象とする被検液は、種々の
試料から調製されたものや、試料自体からなる。
この被検液を構成する溶媒は前述のごとく種々の
ものが挙げられるが、少なくともイオン性物質を
微量（通常、20ppm以下）含有する被検液がこの
発明の対象となる。この例としては、前述のごと
く非水溶性液状媒体中に微量のイオン性物質を含
有してなる試料を親水性有機溶媒と混合した被検
液や、水溶性溶媒中に微量のイオン性物質を含有
してなる被検液等が代表的である。前者の具体例
としては、テレフタル酸ジメチル（非水溶性液状
媒体）中にトルイル酸、フタル酸、イソフタル
酸、テレフタル酸、テレフタル酸モノメチル等の
アニオン性の物質を微量含有してなる試料を、水
と任意に混合する有機溶媒（例えば、メタノー
ル、エタノール、ｎ−プロパノール、ｉ−プロパ
ノール、ｔ−ブタノール、アセトン、ジオキサ
ン、アセトニトリル等）で溶解してなる被検液が
挙げられる。また後者の具体例としては、蒸留水
中の無機イオン、例えば塩素イオン、硝酸イオ
ン、硫酸イオン、ナトリウムイオン、アンモニウ
ムイオン等が挙げられる。 On the other hand, the test liquid that is the subject of this invention is prepared from various samples or consists of the sample itself.
Although various solvents can be used as the solvent constituting this test liquid, as described above, a test liquid containing at least a trace amount (usually 20 ppm or less) of an ionic substance is the object of the present invention. Examples of this include a test solution in which a sample containing a trace amount of an ionic substance in an aqueous non-aqueous liquid medium is mixed with a hydrophilic organic solvent, as described above, and a test solution containing a trace amount of an ionic substance in a water-soluble solvent. A typical example is a test liquid containing such a substance. As a specific example of the former, a sample containing a trace amount of an anionic substance such as toluic acid, phthalic acid, isophthalic acid, terephthalic acid, or monomethyl terephthalate in dimethyl terephthalate (a water-insoluble liquid medium) is mixed with water. Examples include a test liquid dissolved in an organic solvent (for example, methanol, ethanol, n-propanol, i-propanol, t-butanol, acetone, dioxane, acetonitrile, etc.) that is optionally mixed with. Specific examples of the latter include inorganic ions in distilled water, such as chlorine ions, nitrate ions, sulfate ions, sodium ions, and ammonium ions.
上記各被検液中に、前記イオン性化合物を加え
る方法としては、直接所定量のイオン性化合物を
添加する方法、試料を溶解して被検液とする際の
親水性有機溶媒中に予めイオン性化合物を溶解し
ておく方法、イオン性化合物の水溶液を上記被検
液に加えて新たに被検液とする方法等が挙げられ
る。なお、イオン性化合物の水溶液を加えた場合
に非水溶性液状媒体の分相が生じた際には、これ
を除去することが好ましい。 The ionic compound can be added to each of the above test solutions by directly adding a predetermined amount of the ionic compound, or by adding ions in advance to a hydrophilic organic solvent when dissolving the sample to prepare the test solution. Examples include a method in which an ionic compound is dissolved in advance, and a method in which an aqueous solution of an ionic compound is added to the above-mentioned test liquid to prepare a new test liquid. Note that when phase separation of the non-aqueous liquid medium occurs when an aqueous solution of an ionic compound is added, it is preferable to remove this.
上記のごときイオン性化合物を含有する被検液
は等速電気泳動に付されるが、この際用いる装置
や泳動条件自体は当該分野で公知のものを採用す
ることができる。例えば、リーデイング液やター
ミナル液中に被検液との親和性を向上させるため
の親水性有機溶媒が加えられていてもよい。 The test solution containing the above ionic compound is subjected to isokinetic electrophoresis, and the apparatus and electrophoresis conditions used at this time can be those known in the art. For example, a hydrophilic organic solvent may be added to the leading liquid or terminal liquid to improve affinity with the test liquid.
(ホ) 実施例
試料としてテトロンの原料となるテレフタル酸
ジメチルを対象とし、この中に微量（通常、数
10ppmオーダー）含まれるイオン性物質すなわち
トルイル酸、フタル酸、イソフタル酸、テレフタ
ル酸、テレフタル酸モノメチル等のアニオン性の
不純物の分析を等速電気泳動分析によつて行なつ
た。(E) Example A sample of dimethyl terephthalate, which is the raw material for Tetron, is used as a sample.
Anionic impurities such as toluic acid, phthalic acid, isophthalic acid, terephthalic acid, and monomethyl terephthalate were analyzed by isotachophoresis analysis.
まず、上記試料（テレフタル酸ジメチル）0.5
ｇをアセトン５mlに溶解し、これにさらにイミダ
ゾールの10ｍＭ水溶液（この発明におけるイオン
性化合物の水溶液）を加えて、全量10mlとし、生
じた沈殿を分離後、上澄みの10μ及び50μを
細管式等速電気泳動分析装置〔IP2A型、島津製
作所製〕に注入した電気泳動を行なつた。なお、
この場合のリーデング液としては、（0.01M−Ｌ
−ヒスチジン塩酸塩＋0.01M−Ｌ−ヒスチジ
ン）／アセトン＝（１／１）の水性溶液を用い、
ターミナル液としては、0.01Mのｎ−カプロン酸
ナトリウム水溶液を用いた。 First, the above sample (dimethyl terephthalate) 0.5
Dissolve g in 5 ml of acetone, add 10 mM aqueous solution of imidazole (aqueous solution of ionic compound in this invention) to make a total volume of 10 ml, separate the resulting precipitate, and then transfer 10 μ and 50 μ of the supernatant to a capillary tube at constant velocity. Electrophoresis was performed using an electrophoresis analyzer (IP2A type, manufactured by Shimadzu Corporation). In addition,
In this case, the leading liquid is (0.01M-L
- using an aqueous solution of histidine hydrochloride + 0.01M - L-histidine)/acetone = (1/1),
As the terminal liquid, a 0.01M aqueous solution of sodium n-caproate was used.
このようにして得られた分析結果を第１図
（10μ注入）及び第２図（50μ注入）に示し
た。 The analysis results thus obtained are shown in FIG. 1 (10μ injection) and FIG. 2 (50μ injection).
一方、比較例として、イミダゾール水溶液に代
えてリーデング電解液を同容量加え、同様に分析
を行なつた結果を第３図（10μ注入）に示し
た。 On the other hand, as a comparative example, the same volume of leading electrolyte solution was added in place of the imidazole aqueous solution, and the same analysis was carried out. The results are shown in FIG. 3 (10μ injection).
図中、Ａはリーデングイオン（塩素イオン）、
Ｂはフタル酸、Ｃはテレフタル酸モノメチル、Ｄ
はｐ−トルイル酸、Ｅはターミナルイオン（ｎ−
カプロン酸イオン）をそれぞれ示すものである。
なお、比較例の被検液を50μ注入したところ泳
動管内に気泡が発生して通電不可能となつた。 In the figure, A is a leading ion (chlorine ion),
B is phthalic acid, C is monomethyl terephthalate, D
is p-toluic acid, E is terminal ion (n-
caproate ion).
Note that when 50μ of the test liquid of the comparative example was injected, bubbles were generated in the electrophoresis tube, making it impossible to apply electricity.
第１図及び第３図（比較例）に示されるよう
に、微量イオン性物質と逆性のイオン性化合物
（イミダゾール）を加えた被検液を用いた際には、
リーデイングイオン量が少ないために分析時間が
短縮化されていることが判る。また50μの被検
液を注入した場合においても通電性は良好であ
り、被検液の増量に対応して感度が向上している
ことが判る。 As shown in Figures 1 and 3 (comparative examples), when using a test solution containing a trace amount of ionic substance and an opposite ionic compound (imidazole),
It can be seen that the analysis time is shortened due to the small amount of leading ions. Furthermore, even when 50μ of the test liquid was injected, the conductivity was good, and it was found that the sensitivity improved as the amount of the test liquid increased.
(ホ) 発明の効果
この発明の方法によれば、イオン性物質の量が
微量の試料や被検液を、通電性のトラブルを生じ
ることなく短時間で等速電気泳動分析することが
でき、しかも被検液の注入量を増加させることに
より感度の向上も容易に行なうことができる。そ
して、かりにイオン性化合物が加えられた被検液
が泳動用の電解液中で分相を起して目的の微量イ
オンが一時的に共沈した場合においても、水良溶
性のイオン性化合物が加えられているため系の通
電性は実質的に阻害されることなく、微量イオン
の分析を行なうことも可能である。(E) Effects of the Invention According to the method of the invention, samples and test liquids containing minute amounts of ionic substances can be analyzed by isokinetic electrophoresis in a short time without causing problems with electrical conductivity. Moreover, the sensitivity can be easily improved by increasing the injection amount of the test liquid. Even if the sample solution to which an ionic compound is added undergoes phase separation in the electrolyte for electrophoresis and the desired trace ions temporarily co-precipitate, the water-soluble ionic compound Since this is added, the conductivity of the system is not substantially inhibited, and it is also possible to analyze trace ions.
第１図及び第２図は、この発明の方法で得られ
る分析チヤート（電位勾配−時間特性）を例示す
るイソタコフエログラム、第３図は従来の方法で
得られる分析チヤートを例示するイソタコフエロ
グラムである。
1 and 2 are isotacopherograms illustrating an analytical chart (potential gradient-time characteristics) obtained by the method of the present invention, and FIG. 3 is an isotacopherogram illustrating an analytical chart obtained by a conventional method. It is a ferogram.
Claims (1)
被検液を等速電気泳動分析に付して該溶媒中の所
望のイオン性物質を分析するに際し、上記被検液
中に、分析を意図するイオンと逆性のイオンとな
りかつ上記溶媒及び水に良溶性のイオン性化合物
を加えて等速電気泳動を行うにあたつて、分析を
意図するイオンがアニオンの場合、加えるイオン
性化合物がアミン類であり、分析を意図するイオ
ンがカチオンの場合、加えるイオン性化合物が無
機酸又は有機酸であることを特徴とする電気泳動
分析方法。 ２ 被検液が、非水溶性液状媒体中に微量のイオ
ン性物質を含有してなる溶液を親水性有機溶媒と
混合してなり、任意に水を混合した溶液からなる
特許請求の範囲第１項記載の分析方法。 ３ 被検液が、水溶性媒体中に微量のイオン性物
質を含有してなる溶液からなる特許請求の範囲第
１項記載の分析方法。[Scope of Claims] 1. When a test solution containing a trace amount of an ionic substance in a solvent is subjected to isokinetic electrophoresis analysis to analyze a desired ionic substance in the solvent, When performing isokinetic electrophoresis by adding an ionic compound to the solution that has the opposite polarity to the ion intended for analysis and is well soluble in the above solvent and water, if the ion intended for analysis is an anion. An electrophoretic analysis method characterized in that the ionic compound added is an amine, and when the ion intended for analysis is a cation, the ionic compound added is an inorganic acid or an organic acid. 2. Claim 1, in which the test liquid is a solution containing a trace amount of an ionic substance in a non-aqueous liquid medium, mixed with a hydrophilic organic solvent, optionally mixed with water. Analytical method described in section. 3. The analysis method according to claim 1, wherein the test liquid is a solution containing a trace amount of an ionic substance in an aqueous medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60035095A JPS61194342A (en) | 1985-02-22 | 1985-02-22 | Electrophoretic analysis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60035095A JPS61194342A (en) | 1985-02-22 | 1985-02-22 | Electrophoretic analysis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS61194342A JPS61194342A (en) | 1986-08-28 |
| JPH0535825B2 true JPH0535825B2 (en) | 1993-05-27 |
Family
ID=12432383
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60035095A Granted JPS61194342A (en) | 1985-02-22 | 1985-02-22 | Electrophoretic analysis |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS61194342A (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9104411D0 (en) * | 1991-03-01 | 1991-04-17 | Elchrom Limited | Improvement of gel composition in gels for submerged gel electrophoresis |
| JP2800754B2 (en) * | 1996-01-12 | 1998-09-21 | 日本電気株式会社 | Method for analyzing ions in liquid crystals |
| JP2014055979A (en) * | 2013-12-09 | 2014-03-27 | Sharp Corp | Electric field generation device and electric field generation method |
| JP2020060534A (en) * | 2018-10-12 | 2020-04-16 | 国立大学法人福井大学 | Anion detection method and kit therefor |
-
1985
- 1985-02-22 JP JP60035095A patent/JPS61194342A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS61194342A (en) | 1986-08-28 |
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