JPH0715467B2 - Anion analysis method - Google Patents
Anion analysis methodInfo
- Publication number
- JPH0715467B2 JPH0715467B2 JP60067411A JP6741185A JPH0715467B2 JP H0715467 B2 JPH0715467 B2 JP H0715467B2 JP 60067411 A JP60067411 A JP 60067411A JP 6741185 A JP6741185 A JP 6741185A JP H0715467 B2 JPH0715467 B2 JP H0715467B2
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- JP
- Japan
- Prior art keywords
- ion
- anion
- anions
- chromatography
- phase
- 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
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (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、液体クロマトグラフイによる陰イオン分析
法に関する。さらに詳しくは逆相イオンペアクロマトグ
ラフイによる陰イオン分析の改良に関する。The present invention relates to an anion analysis method by liquid chromatography. More specifically, it relates to improvement of anion analysis by reverse phase ion pair chromatography.
（ロ）従来技術 液体クロマトグラフイによる陰イオンの分析法としては
いわゆるイオンクロマトグラフイ法が知られているが、
装置が専用化してしまい汎用性に乏しくかつ高価であ
る。この点に鑑み、イオン交換クロマトグラフイにおい
て、移相中に紫外吸収性イオンを添加して陰イオンの分
離を行ない、対象陰イオンの溶出の際にこの紫外吸収性
イオンが減少して溶出液の紫外吸光光度が低下する現象
を利用して各種陰イオンの分離分析を行なう提案がなさ
れている（特開昭57-20662号公報）。この方法は分析対
象の陰イオンと上記光吸収性イオン（陰イオン）との和
が系中の対イオン（陽イオン）に対応して一定となる電
気的中性の原理に基づくものであり、光吸収性イオンは
陰イオンの溶離用として働きかつ開接的な陰イオンの検
出用として働く。そして液体クロマトグラフイで一般の
用いられる紫外吸光光度計を用いるため汎用性は高い方
法といえる。(B) Conventional technology As a method for analyzing anions by liquid chromatography, a so-called ion chromatography method is known.
The device is dedicated and has poor versatility and is expensive. In view of this point, in ion exchange chromatography, ultraviolet absorbing ions are added during phase shift to separate anions, and when the target anions are eluted, the ultraviolet absorbing ions decrease and the eluate A proposal has been made to separate and analyze various anions by utilizing the phenomenon that the ultraviolet absorption intensity of the above-mentioned decreases (Japanese Patent Laid-Open No. 57-20662). This method is based on the principle of electrical neutrality in which the sum of the anion to be analyzed and the light absorbing ion (anion) is constant corresponding to the counter ion (cation) in the system, The light-absorbing ion serves as an elution of anion and serves as a detection of an anion that is cleaved. It can be said that the method is highly versatile because it uses an ultraviolet absorptiometer commonly used in liquid chromatography.
かかる紫外吸収性溶離イオンを用いた手法を、いわゆる
逆相イオンペアクロマトグラフイに応用する提案もなさ
れている〔M.Cooke,“Journal of High Resolution Chr
omatography ＆ Chromatography Communications"Vol7,
第515〜519頁（1984）〕。ここで逆相イオンペアクロマ
トグラフイとは、極性移動相と非極性固定相とを組み合
せ、分析対象のイオンとイオンペアを形成しうるカウン
ターイオンの存在下で分離分析を行なう方法であり、一
般にイオン交換クロマトグラフイよりも高カラム効率が
得られ、またイオン交換クロマトグラフイでの成分の保
持は充填剤のイオン交換容量により決定されるのに対
し、この逆相イオンペアクロマトグラフイでは極性移動
相に添加するカウンターイオン（例えば、テトラブチル
アンモニウムイオン）の濃度により保持を調節するとい
う利点を備えた方法である。そして上記M.Cookeの文献
によれば、カウンターイオンとしてテトラブチルアンモ
ニウムを用い、紫外吸収性溶離イオンとしてフタル酸を
用いることにより各種陰イオンの紫外吸収による分離分
析が可能であるとされている。Proposals have also been made to apply the technique using such ultraviolet absorbing eluting ions to so-called reverse-phase ion pair chromatography [M. Cooke, “Journal of High Resolution Chr.
omatography & Chromatography Communications "Vol7,
515-519 (1984)]. Reverse-phase ion-pair chromatography is a method in which a polar mobile phase and a non-polar stationary phase are combined, and separation analysis is performed in the presence of counter ions that can form an ion pair with the ion to be analyzed. The column efficiency is higher than that of chromatography, and the retention of components in ion exchange chromatography is determined by the ion exchange capacity of the packing material. This is a method with the advantage that retention is controlled by the concentration of the counter ion (eg, tetrabutylammonium ion) added. According to the M. Cooke document, it is possible to separate and analyze various anions by ultraviolet absorption by using tetrabutylammonium as a counter ion and phthalic acid as an ultraviolet absorbing elution ion.
しかしながら、上記逆相イオンペアクロマトグラフイー
においては、移動相中に添加される紫外吸収性溶離イオ
ン、すなわちフタル酸の濃度があまり高いと吸光度測定
セルを通過する光量自体が小さくなり、その結果バツク
グラウンドが高くなつてノイズやドリフトの原因とな
り、また直線性も悪化するという問題点が生じる。その
ためフタル酸添加量をできるだけ減少させることが考え
られるが、この場合には保持時間が長くなつて感度が低
下し、ことに硫酸イオンなどの保持の強い陰イオンを含
む試料を対象とした際に分析時間が著しく長くなるとい
う不都合が生じる。また、移動相のpHを上げることによ
り硫酸イオンなどの溶出をはやめることも考えられる
が、溶出のはやいイオンの分離を損わずに行うには限界
がある。さらにまた、pHを上げることは液体クロマトグ
ラフイで広く用いられるシリカ担体の逆相クロマトグラ
フイ用充てん剤の劣化をはやめることにもなり好ましく
ない。However, in the above-mentioned reversed-phase ion pair chromatography, if the concentration of the ultraviolet-absorbing eluting ions added to the mobile phase, that is, phthalic acid is too high, the light amount itself passing through the absorbance measuring cell becomes small, and as a result the background As the value becomes higher, it causes noise and drift, and linearity deteriorates. Therefore, it is possible to reduce the amount of phthalic acid added as much as possible, but in this case, the retention time becomes longer and the sensitivity decreases, especially when a sample containing strongly anions such as sulfate ions is targeted. The disadvantage is that the analysis time becomes extremely long. Further, it is possible to stop the elution of sulfate ions and the like by increasing the pH of the mobile phase, but there is a limit to carrying out the separation of ions that elute quickly without impairing them. Furthermore, increasing the pH is also not preferable because it deteriorates the deterioration of the packing material for reverse phase chromatography of the silica carrier which is widely used in liquid chromatography.
（ハ）発明の目的 この発明は、上記従来の問題点を解消すべくなされたも
のであり、感度の低下や保持時間の増加を招くことな
く、ノイズやドリフトを改善できる紫外吸収による逆相
イオンペアクロマトグラフイを提供しようとするもので
ある。(C) Object of the invention The present invention has been made to solve the above-mentioned conventional problems, and it is possible to improve the noise and the drift without reducing the sensitivity and increasing the holding time. It is intended to provide a chromatography.
本発明者らは、各種分析条件について鋭意研究を行なつ
た結果、紫外吸収性溶離イオンとしてフタル酸の代わり
にトリメシン酸等の芳香族三塩基酸を用いることによ
り、フタル酸より低い添加量で、かつ、pHを上げること
なく弱酸性領域で陰イオンの保持時間を短縮でき、陰イ
オンの保持時間を短縮でき、溶出のはやいイオンの分離
を損わずに硫酸イオン等の保持の強い陰イオンの溶出時
間を著しく短縮化でき、またフタル酸よりも低い添加量
でも従来と同程度の分析時間や感度が得られるためセル
通過光量を分析時間や感度に悪影響を与えることなく増
加させることができバツクグラウンドを低減させること
ができる事実を見出した。As a result of intensive studies on various analysis conditions, the present inventors have used an aromatic tribasic acid such as trimesic acid instead of phthalic acid as an ultraviolet-absorbing eluting ion, thereby adding a lower amount than phthalic acid. In addition, the retention time of anions can be shortened in a weakly acidic region without raising the pH, the retention time of anions can be shortened, and anions with strong retention of sulfate ions without impairing the separation of ions that elute quickly The elution time can be remarkably shortened, and the amount of light passing through the cell can be increased without adversely affecting the analysis time or sensitivity because the same analysis time and sensitivity as before can be obtained with a lower addition amount than phthalic acid. We have found the fact that the background can be reduced.
（ニ）発明の構成 かくしてこの発明によれば、極性移動相と非極性固定相
とを用い分析対象の陰イオンとイオンペアを形成しうる
カウンターイオンの存在下で陰イオンの分離分析を行な
う逆相イオンペアクロマトグラフイにおいて、 上記極性移動相中に紫外吸光性溶離剤としての芳香族三
塩基酸を添加して陰イオンの分離を行ない、分離流出液
の芳香族三塩基酸による紫外吸光光度に基づいて陰イオ
ンを検出することを特徴とする陰イオン分析法が提供さ
れる。(D) Structure of the Invention Thus, according to the present invention, a reversed phase in which an anion is separated and analyzed using a polar mobile phase and a non-polar stationary phase in the presence of a counter ion capable of forming an ion pair with the anion to be analyzed. In ion pair chromatography, aromatic tribasic acid as an ultraviolet light absorbing eluent was added to the polar mobile phase to separate anions, and the separation effluent was analyzed based on the ultraviolet absorptivity of the aromatic tribasic acid. An anion analysis method is provided, which comprises detecting anions.
この発明は、逆相イオンペアクロマトグラフイにおける
従来の紫外吸収性溶離イオンの代わりに芳香族三塩基酸
を用いたものである。これにより従来に比して分析時間
を改善することができ、またバツクグラウンドを低下さ
せることができるため、ノイズ、ドリフト、感度及び直
線性を改善することが可能である。This invention uses an aromatic tribasic acid in place of the conventional ultraviolet absorbing eluent ion in reversed phase ion pair chromatography. As a result, the analysis time can be improved and the background can be reduced as compared with the conventional case, so that noise, drift, sensitivity and linearity can be improved.
上記芳香族三塩基酸としては、トリメシン酸、トリメリ
ツト酸、ヘミメリツト酸等のベンゼントリカルボン酸が
挙げられる。かかる芳香族三塩基酸はもちろん塩の形態
で添加されてもよい。この添加量は条件によつても異な
るが、紫外光の検出器の感度やバツクグラウンドの点
で、0.1〜1.5mM（移動相中）とするのが適している。Examples of the aromatic tribasic acid include benzenetricarboxylic acid such as trimesic acid, trimellitic acid and hemimellitic acid. Such aromatic tribasic acids may of course be added in salt form. Although the added amount varies depending on the conditions, it is suitable to be 0.1 to 1.5 mM (in the mobile phase) in view of the sensitivity of the detector of ultraviolet light and the background.
なお、他の条件は公知の逆相イオンペアクロマトグラフ
イで知られた条件を適宜選択すればよい。その代表例と
しては、いわゆるODS系の非極性固定相を用い、カウン
ターイオンとしてのテトラブチルアンモニウムイオンを
含むpH4〜７の移動相を基本的に用いて溶離を行なう方
法が挙げられる。Note that other conditions may be appropriately selected from those known by known reversed-phase ion pair chromatography. As a typical example thereof, there is a method in which a so-called ODS-based non-polar stationary phase is used, and elution is basically performed using a mobile phase having a pH 4 to 7 containing tetrabutylammonium ion as a counter ion.
（ホ）実施例 各種陰イオン成分を含む試料（水溶液）についてこの発
明の方法により分離分析を行なつた。用いた試料中の陰
イオン成分は下表の通りである。(E) Example A sample (aqueous solution) containing various anion components was separated and analyzed by the method of the present invention. The anion components in the samples used are as shown in the table below.
なお、分析条件は以下の通りである。 The analysis conditions are as follows.
分析カラム:Shim−pack CLC−ODS カラム （島津製作所社製；内径6.0mm長さ15cm） 移動相:0.8mMトリメシン酸（紫外吸光性溶離剤）、1.0m
M水酸化テトラブチルアンモニウム、（カウンターイオ
ン）を含むpH4.6水溶液（KOHでpH調整） 流 速:1.5ml/分 カラム温度:40℃ 検出：紫外282nm（感度0.08AUFS） 試料注入量:20μｌ 得られた結果を第１表に示す。なお、図中の番号は前記
表１のNo.に対応する。このように各陰イオン成分が保
持時間約10分で効率良く分離分析されていることが判
る。Analytical column: Shim-pack CLC-ODS column (manufactured by Shimadzu Corporation; inner diameter 6.0 mm, length 15 cm) Mobile phase: 0.8 mM trimesic acid (ultraviolet absorbing eluent), 1.0 m
PH 4.6 aqueous solution containing M tetrabutylammonium hydroxide (counter ion) (pH adjusted with KOH) Flow rate: 1.5 ml / min Column temperature: 40 ° C Detection: UV 282 nm (sensitivity 0.08 AUFS) Sample injection volume: 20 μl Obtained The results obtained are shown in Table 1. The numbers in the figure correspond to the numbers in Table 1 above. Thus, it can be seen that each anion component is efficiently separated and analyzed with a retention time of about 10 minutes.
これに対し、トリメシン酸の代わりにフタル酸を用いて
上記と同条件下で分析を行なつた場合には、硫酸イオン
の溶出に30分以上かかることとなり、分析時間を短縮し
感度を高めるためには、フタル酸の濃度を増加させる必
要があることが判るが、この場合にはバツクグラウンド
を上昇させることとなるため好ましくない。On the other hand, if phthalic acid was used instead of trimesic acid and the analysis was performed under the same conditions as above, it would take 30 minutes or more to elute the sulfate ion, which shortened the analysis time and increased the sensitivity. , It is necessary to increase the concentration of phthalic acid, but in this case, the background is increased, which is not preferable.
（ヘ）発明の効果 この発明の陰イオン分析法によれば、フタル酸の代わり
に陰イオン溶出力の高い芳香族塩基酸を紫外吸光性溶離
剤として用いるため、少ない添加量で各種陰イオンの分
離を効率良く行なうことができる。そのため、従来に比
してバツクグラウンドを減少させることができ、ノイズ
やドリフトを低減でき、濃度−感度の直線性も改善する
ことができる。(F) Effect of the Invention According to the anion analysis method of the present invention, since an aromatic basic acid having a high anion-dissolving output is used as an ultraviolet absorptive eluent instead of phthalic acid, a small amount of various anions can be added. Separation can be performed efficiently. Therefore, the background can be reduced, noise and drift can be reduced, and the concentration-sensitivity linearity can be improved as compared with the conventional case.
第１図はこの発明の陰イオン分析法により得られる分析
結果を例示するクロマトグラム図である。FIG. 1 is a chromatogram diagram illustrating an analysis result obtained by the anion analysis method of the present invention.
フロントページの続き (56)参考文献 特開 昭57−20662（ＪＰ，Ａ) Ｍ．Ｃｏｏｋｅ，“Ｊｏｕｒｎａｌ ｏ ｆ Ｈｉｇｈ Ｒｅｓｏｌｗｔｉｏｎ Ｃ ｈｒｏｍａｔｏｇｒａｐｈｙ ＆ Ｃｈｒ ｏｍａｔｏｇｒａｐｈｙ Ｃｏｍｍｕｎｉ ｃａｔｉｏｎｓ”，ｖｏｌ．７，Ｐ．515 〜Ｐ．519（1984)Continuation of front page (56) References JP-A-57-20662 (JP, A) M.M. Cooke, "Journal of High Resolution Chromatography & Chromatography Communications", vol. 7, P.I. 515-P. 519 (1984)
Claims (1)
象の陰イオンとイオンペアを形成しうるカウンターイオ
ンの存在下で陰イオンの分離分析を行なう逆相イオンペ
アクロマトグラフィにおいて、 上記極性移動相中に紫外吸光性溶離剤としての芳香族三
塩基酸を添加して弱酸性領域で陰イオンの分離を行い、
分離流出液の芳香族三塩基酸による紫外吸光光度に基づ
いて陰イオンを検出することを特徴とする陰イオン分析
法。1. A reversed-phase ion pair chromatography for separating and analyzing anions using a polar mobile phase and a non-polar stationary phase in the presence of a counter ion capable of forming an ion pair with an anion to be analyzed. Aromatic tribasic acid as an ultraviolet absorbing eluent was added to the mixture to separate anions in the weakly acidic region.
An anion analysis method characterized by detecting anions based on the ultraviolet absorption spectrophotometry of the separated effluent with an aromatic tribasic acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60067411A JPH0715467B2 (en) | 1985-03-30 | 1985-03-30 | Anion analysis method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60067411A JPH0715467B2 (en) | 1985-03-30 | 1985-03-30 | Anion analysis method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS61226658A JPS61226658A (en) | 1986-10-08 |
| JPH0715467B2 true JPH0715467B2 (en) | 1995-02-22 |
Family
ID=13344145
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60067411A Expired - Fee Related JPH0715467B2 (en) | 1985-03-30 | 1985-03-30 | Anion analysis method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0715467B2 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8742331B2 (en) | 2008-02-15 | 2014-06-03 | Sigma-Aldrich, Co. | Imidazolium-based liquid salts and methods of use thereof |
| EP2245653A1 (en) | 2008-02-15 | 2010-11-03 | Sigma-Aldrich Co. | Anion detection by esi-ms using dicationic liquid salts |
| JP2020060534A (en) * | 2018-10-12 | 2020-04-16 | 国立大学法人福井大学 | Anion detection method and kit therefor |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1168705A (en) * | 1980-05-27 | 1984-06-05 | Hamish Small | Ion exchange chromatography with indirect photometric detection |
-
1985
- 1985-03-30 JP JP60067411A patent/JPH0715467B2/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| Ｍ．Ｃｏｏｋｅ，"ＪｏｕｒｎａｌｏｆＨｉｇｈＲｅｓｏｌｗｔｉｏｎＣｈｒｏｍａｔｏｇｒａｐｈｙ＆ＣｈｒｏｍａｔｏｇｒａｐｈｙＣｏｍｍｕｎｉｃａｔｉｏｎｓ"，ｖｏｌ．７，Ｐ．５１５〜Ｐ．５１９（１９８４） |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS61226658A (en) | 1986-10-08 |
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