JPH06138110A - Separator column for ion chromatography - Google Patents
Separator column for ion chromatographyInfo
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- JPH06138110A JPH06138110A JP4289110A JP28911092A JPH06138110A JP H06138110 A JPH06138110 A JP H06138110A JP 4289110 A JP4289110 A JP 4289110A JP 28911092 A JP28911092 A JP 28911092A JP H06138110 A JPH06138110 A JP H06138110A
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Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、イオンクロマトグラフ
ィーに使用する分離カラムに関する。特に、生体関連物
質の分離・分析に好適なものである。FIELD OF THE INVENTION The present invention relates to a separation column used for ion chromatography. In particular, it is suitable for separation / analysis of bio-related substances.
【０００２】以下、本明細書で使用する主たる略語・用
語の一覧を下記する。The following is a list of main abbreviations and terms used in this specification.
【０００３】ＯＤＳ…オクタデシルシリル化ゲル ツビッタ（Zwitter ）…プラス電荷（＋δ）とマイナス
電荷（−δ）をそれぞれ自らに有する化合物。ODS ... Octadecylsilylated gel Zwitter ... A compound having a positive charge (+ δ) and a negative charge (-δ).
【０００４】水系溶媒…水と極性有機溶媒との混合溶
媒、及び水系緩衝液を含む。Aqueous solvent: Contains a mixed solvent of water and a polar organic solvent, and an aqueous buffer solution.
【０００５】[0005]
【従来の技術及び発明が解決しようとする課題】従来の
液体クロマトグラフィーは、分配系の高速液体クロマト
グラフィーが主流で、、さらに、測定できる試料の種類
を増やすためにイオン交換型のクロマトグラフィーも開
発され、普及してきている。2. Description of the Related Art Conventional liquid chromatography is predominantly distributed high-performance liquid chromatography, and ion-exchange type chromatography is also used to increase the number of measurable samples. It has been developed and is becoming popular.
【０００６】しかし、溶離液が水系溶媒のみで、陽イオ
ン、陰イオン、及び陰陽両イオンが同時に、分離できる
方法は、本発明者が知る限りにおいては存在しない。However, as far as the present inventor knows, there is no method capable of simultaneously separating cations, anions, and both anions and cations with an eluent containing only an aqueous solvent.
【０００７】本発明は、上記にかんがみて、水または水
系溶媒のみで、陽イオン、陰イオン、及び陰陽両イオン
が同時に分離できるイオンクロマトグラフィー用分離カ
ラムを提供することにある。In view of the above, the present invention provides a separation column for ion chromatography capable of simultaneously separating cations, anions, and both anions and cations using only water or an aqueous solvent.
【０００８】[0008]
【課題を解決するための手段】本発明のは、上記課題を
下記構成により解決するものである。According to the present invention, the above-mentioned problems are solved by the following constitution.
【０００９】イオンクロマトグラフィーに使用する分離
カラムであって、固定相が、保持体表面にツビッタを結
合被覆させた充填剤で形成され、溶離液として、水また
は水系溶媒を使用することを特徴とする。A separation column used in ion chromatography, characterized in that the stationary phase is formed of a packing material in which the surface of a support is bound and coated with Zwitter, and water or an aqueous solvent is used as an eluent. To do.
【００１０】[0010]
【実施例】以下、本発明を、詳細に説明をする。The present invention will be described in detail below.
【００１１】Ａ．本発明のイオンクロマトグラフィー用
分離カラムは、下記のようにして製造する。A. The separation column for ion chromatography of the present invention is manufactured as follows.
【００１２】(1) 保持体（被修飾用充填剤）を充填した
逆相液体クロマトグラフィー（ＲＰＬＣ）用分離カラム
の中に、ツビッタ飽和水系溶液を流し、上記保持体表面
に、ツビッタを結合被覆（疎水結合、できれば化学結
合）させる。(1) A Zitterta saturated aqueous solution is caused to flow into a separation column for reverse phase liquid chromatography (RPLC) packed with a support (a packing material to be modified), and the surface of the support is bound and coated with Zwitter. (Hydrophobic bond, preferably chemical bond).
【００１３】上記被修飾充填剤としては、代表的にＯ
ＤＳシリカゲル等を挙げることができる。ＯＤＳ（炭素
数１８）以外に類似の構造を有するものも使用可能であ
るが、炭素数が１０以下のように小さくなると、疎水性
が小さくなり、被覆力（疎水結合性）が小さくなり、溶
離液として水系溶媒を使用した場合、脱離し易く、耐久
性に問題が発生し易くなる。As the above-mentioned modified filler, O is typical.
DS silica gel etc. can be mentioned. Other than ODS (having 18 carbon atoms), those having a similar structure can also be used, but when the carbon number becomes 10 or less, the hydrophobicity becomes small, the covering power (hydrophobic binding property) becomes small, and the elution occurs. When an aqueous solvent is used as the liquid, it is likely to be desorbed and a durability problem is likely to occur.
【００１４】上記ツビッタとしては、被修飾用充填剤
（保持体）の表面を結合被覆可能なものなら、特に限定
されないが、コール酸、デキシコール酸、グリココール
酸、デキシグリココール酸、タウロコール酸、デオキシ
タウロコール酸、等の胆汁酸の金属塩類を例示でき、好
適には、下記化学構造式で示されるものを使用できる。The Zwitter is not particularly limited as long as it can bind and coat the surface of the filler (support) to be modified, and is not particularly limited. Examples thereof include metal salts of bile acids such as deoxytaurocholic acid, and preferably those represented by the following chemical structural formula can be used.
【００１５】[0015]
【化１】 [Chemical 1]
【００１６】[0016]
【化２】 [Chemical 2]
【００１７】[0017]
【化３】 [Chemical 3]
【００１８】ツビッタ飽和水系溶液としては、ツビッ
タが、ＣＨＡＰＳの場合、例えば水／アセトニトリル
（容量比）＝１／１の水系溶媒を使用する。If CHAPS is used as the saturated aqueous solution of Twitter, for example, an aqueous solvent of water / acetonitrile (volume ratio) = 1/1 is used.
【００１９】(2) 次に、蒸留水を、上記分離カラム内を
通して水洗した後、緩衝液を、同様にして上記カラム内
を通して、ツビッタの膜厚を調製する。(2) Next, distilled water is passed through the separation column and washed with water, and then a buffer solution is passed through the column in the same manner to adjust the film thickness of the Zitter.
【００２０】上記、緩衝液としては、例えば、リン酸塩
水溶液を好適に使用可能である。 Ｂ．上記のようにして調製した本発明の分離カラムは、
例えば、図１に示すようなクロマトグラフィー装置に、
組み込んで使用する。As the above buffer solution, for example, a phosphate aqueous solution can be preferably used. B. The separation column of the present invention prepared as described above,
For example, in a chromatography device as shown in FIG.
Used by incorporating.
【００２１】装置の概略は、溶離液タンク１１と、送液
ポンプ１３、試料導入部１５、分離カラム１７、検出器
１９とを備え、溶離液タンク１１と検出器１９との間
は、被検出成分の拡散を可及的に小さく維持できるよう
に、接続されている。なお、検出器１９は、増幅器２１
を介して表示装置（記録計）２３に接続される。The apparatus is generally provided with an eluent tank 11, a liquid feed pump 13, a sample introduction section 15, a separation column 17, and a detector 19. Between the eluent tank 11 and the detector 19, a substance to be detected is detected. It is connected so that the diffusion of the components can be kept as small as possible. In addition, the detector 19 includes an amplifier 21.
Is connected to the display device (recorder) 23 via.
【００２２】なお、上記検出器としては、イオン成分を
検出できるものなら、特に限定されない。例えば、伝導
度計、分光光度計（紫外・可視）、蛍光光度計、示差屈
折計、質量分析計等を挙げることができる。The detector is not particularly limited as long as it can detect an ionic component. For example, a conductivity meter, a spectrophotometer (ultraviolet / visible), a fluorometer, a differential refractometer, a mass spectrometer, etc. can be mentioned.
【００２３】そして、生体関連物質等の試料の測定操作
は、一般的なイオンクロマトグラフィーと同様の方法に
より行なう。Then, the measurement operation of the sample such as a biological substance is carried out by the same method as the general ion chromatography.
【００２４】(1) まず、バイパス状態の試料導入部１５
のループインジェクタに、試料（被検体）を所定量保持
する。(1) First, the sample introducing section 15 in the bypass state
A predetermined amount of sample (subject) is held in the loop injector.
【００２５】(2) 次に、試料導入部１５を導通状態と
し、試料導入部１５内の試料を、溶離液により分離カラ
ム１７内に展開させ、試料の分離・分析を行なう。(2) Next, the sample introducing unit 15 is brought into conduction, and the sample in the sample introducing unit 15 is developed in the separation column 17 by the eluent to separate and analyze the sample.
【００２６】ここで、溶離液としては、水（通常、蒸留
水）のみでも良いが、水に極性溶剤やリン酸塩緩衝液を
加えて、分離性能を向上させることも可能である。Here, the eluent may be water (usually distilled water) alone, but it is also possible to improve the separation performance by adding a polar solvent or a phosphate buffer solution to water.
【００２７】(3) そして、分離された成分は、検出器１
９に入り検出され、検出結果が、記録計２３に表示され
る。(3) The separated components are detected by the detector 1
9 is detected and the detection result is displayed on the recorder 23.
【００２８】なお、高い塩濃度が必要とされる場合、ま
たは、高いｐＨを必要とされる場合においても、本発明
の固定相は十分な安定性を示す。後述の試験例は、ｐＨ
約１〜１０で十分な安定性示す。The stationary phase of the present invention exhibits sufficient stability even when a high salt concentration is required or a high pH is required. The test examples described below are for pH
About 1 to 10 shows sufficient stability.
【００２９】[0029]
【発明の作用・効果】本発明のイオンクロマトグラフィ
ー用分離カラムは、固定相が保持体表面にツビッタを被
覆させた充填剤であり、溶離液として、水または水系溶
媒を使用することにより、下記のような作用・効果を奏
する。In the ion chromatography separation column of the present invention, the stationary phase is a packing material in which the surface of the support is coated with Zvitta, and by using water or an aqueous solvent as an eluent, It has the same action and effect.
【００３０】生体関連物質（例えば、唾液・尿・血清
等）等の試料を分離カラム内に導入した場合、試料中に
イオン化していないタンパク質等の有機体が含まれてい
ても、それらはほとんど、固定相と相互作用することな
く、システムピークとして表れ、試料中の目的イオン分
離の妨害とはならない。固定相（ツビッタ）と、試料イ
オンとの相互作用が、静電的な吸引・反発力に基づくた
めである。このため、生体関連物質を試料とする場合に
おいて、タンパク質除去等の前処理が不要となる。When a sample such as a bio-related substance (eg saliva, urine, serum, etc.) is introduced into a separation column, even if the sample contains organisms such as unionized proteins, most of them are not present. , Appears as a system peak without interacting with the stationary phase, and does not interfere with the separation of target ions in the sample. This is because the interaction between the stationary phase (Zwitter) and the sample ions is based on electrostatic attraction / repulsion. Therefore, when using a biological substance as a sample, pretreatment such as protein removal is unnecessary.
【００３１】そして、溶離液として水のみでイオンクロ
マトグラフィーを行なうことができる、即ち、従来のイ
オンクロマトグラフィーの如く、溶離液中に緩衝剤等を
加えずに行なえる。Ion chromatography can be carried out only with water as the eluent, that is, like conventional ion chromatography, it can be carried out without adding a buffer or the like to the eluent.
【００３２】従って、上記前処理を不要で、かつ、水の
みを溶離液としてイオンクロマトグラフィーを行なうこ
とができるので、試料中の微量な分析目的物でも、その
逸散・破壊・汚染がほとんどなく、環境中の存在状態に
近い状態で、分析可能となる。Therefore, since the above-mentioned pretreatment is unnecessary and the ion chromatography can be carried out using only water as an eluent, even a trace amount of the objective substance to be analyzed is hardly scattered, destroyed or contaminated. , It becomes possible to analyze in a state close to the existing state in the environment.
【００３３】この分離カラムを使用してイオンクロマト
グラフィーを、臨床分析、環境分析さらには、天然物化
学分野の分析に応用すれば、従来では困難であった正確
な試料の持つ情報を引き出すことが期待できるものであ
る。If ion chromatography is applied to clinical analysis, environmental analysis, and analysis in the field of natural product chemistry using this separation column, it is possible to extract accurate information possessed by an accurate sample, which was difficult in the past. It can be expected.
【００３４】なお、液体クロマトグラフィーとマススペ
クトロメトリーとの連結において、溶離液が水のみとい
うことは脱溶媒部が無用となる、という効果も奏する。In the connection of liquid chromatography and mass spectrometry, the fact that the eluent is only water has an effect that the desolvation part becomes unnecessary.
【００３５】[0035]
【試験例】以下、本発明の効果を確認するために行なっ
た試験例について説明をする。[Test Example] Hereinafter, a test example performed for confirming the effect of the present invention will be described.
【００３６】(i) クロマトグラフィー用カラムの調製 市販のＯＤＳ充填カラム内に、下記ツビッタを、水／ア
セトニトリル系溶媒に１００ｍモルの濃度で溶解し、
０．５ｍｌ／min の速度で約２０分間流下させて、ＯＤ
Ｓ充填剤（保持体）の表面にツビッタ膜（ミセル膜）を
疎水結合により被覆させる。(I) Preparation of Chromatographic Column In a commercially available ODS packed column, the following Zwitter was dissolved in a water / acetonitrile-based solvent at a concentration of 100 mmol,
Flow it down at a rate of 0.5 ml / min for about 20 minutes, and
The surface of the S filler (support) is coated with a vititter film (micelle film) by hydrophobic bonding.
【００３７】その後、被覆時と同様の条件で蒸留水によ
り水洗を行ない、さらには、１ｍmol リン酸緩衝液を、
同様の条件で流し、膜厚調製を行なう。Thereafter, washing with distilled water was carried out under the same conditions as the coating, and further 1 mMol phosphate buffer solution was added.
Flow under the same conditions to adjust the film thickness.
【００３８】ツビッタ…試験例１：ＣＨＡＰＳ、試験例
２：ＮａＴＤＣ、いずれもシグマ社製 (ii)試料の分離方法 上記カラムの流出口を、検出器（紫外線分光光度計：
「ＰＣＤ−６Ａ」、電導度検出器：「ＣＤＤ６Ａ」、島
津製作所社製）に接続する。Zwitter: Test Example 1: CHAPS, Test Example 2: NaTDC, both manufactured by Sigma (ii) Sample separation method The outlet of the above column was connected to a detector (UV spectrophotometer:
"PCD-6A", conductivity detector: "CDD6A", manufactured by Shimadzu Corporation).
【００３９】試料はそれぞれ、下記処方のものを使用
した。The samples used had the following formulations.
【００４０】試験例１（陰イオン含有水溶液）：Ｉ
Ｏ3 -、ＮＯ2 -、Ｎ０3 -、ＳＣＮ- の標準各０．５ｍモル
溶液 試験例２（陰陽両イオン含有水溶液）：Ｉ- 、ＳＣＮ
- 、Ｎａ+ 、Ｋ+ の標準各１０ｍモル溶液 次に、上記試料１０μｌを移動相流（バルブインジェ
クタ）によりカラム内へ流入させる。そして、溶離液と
して下記のものを使用して、下記流速で流下させて、上
記試料の分離を行った。そして、それぞれ下記検出波長
で検出した。Test Example 1 (anion-containing aqueous solution): I
O 3 − , NO 2 − , N0 3 − , SCN − standard 0.5 mmol each solution Test Example 2 (anion and cation ion containing aqueous solution): I − , SCN
- , Na + , K + standard 10 mmol each solution Next, 10 μl of the sample is introduced into the column by a mobile phase flow (valve injector). Then, the following was used as an eluent, and the sample was separated at the following flow rate. Then, each was detected at the following detection wavelengths.
【００４１】 試験例１…水、１ｍｌ／min ；ＵＶ２３０ｎｍ 試験例２…２．５×１０-3モル硫酸銅水溶液、１ｍｌ／
min ； ＵＶ２１５ｎｍ (iii) 結果及び考察 上記試験結果を、図２（試験例１）及び図３（試験例
２）に示す。奇麗なピークが表れ、分離効率が良好であ
ることが分る。Test Example 1 ... Water, 1 ml / min; UV 230 nm Test Example 2 ... 2.5 × 10 −3 mol copper sulfate aqueous solution, 1 ml / min
min; UV215nm (iii) Results and Discussion The test results are shown in Fig. 2 (Test Example 1) and Fig. 3 (Test Example 2). It can be seen that a beautiful peak appears and the separation efficiency is good.
【図１】本発明の分離カラムの適用するイオクロマトグ
ラフィー装置の擬略図である。FIG. 1 is a pseudo-schematic diagram of an ion chromatography device to which the separation column of the present invention is applied.
【図２】試験例１（陰イオン試料）の分析結果を示すク
ロマトグラムである。FIG. 2 is a chromatogram showing the analysis results of Test Example 1 (anion sample).
【図３】試験例２（陰陽両イオン試料）の分析結果を示
すクロマトグラムである。FIG. 3 is a chromatogram showing the analysis results of Test Example 2 (anion and cation sample).
１１ 溶離液タンク １５ 試料導入部 １７ 分離カラム １９ 検出器 11 Eluent tank 15 Sample introduction part 17 Separation column 19 Detector
Claims (1)
離カラムであって、 固定相が、保持体表面にツビッタを結合被覆させた充填
剤で形成され、 溶離液として、水または水系溶媒を使用することを特徴
とする、イオンクロマトグラフィー用分離カラム。1. A separation column for use in ion chromatography, wherein the stationary phase is formed of a packing material in which the surface of a support is bound and coated with Zwitter, and water or an aqueous solvent is used as an eluent. Characteristic separation column for ion chromatography.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4289110A JPH06138110A (en) | 1992-10-27 | 1992-10-27 | Separator column for ion chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4289110A JPH06138110A (en) | 1992-10-27 | 1992-10-27 | Separator column for ion chromatography |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH06138110A true JPH06138110A (en) | 1994-05-20 |
Family
ID=17738921
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4289110A Withdrawn JPH06138110A (en) | 1992-10-27 | 1992-10-27 | Separator column for ion chromatography |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH06138110A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010514674A (en) * | 2006-10-12 | 2010-05-06 | ベラス ヘルス （インターナショナル） リミテッド | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
-
1992
- 1992-10-27 JP JP4289110A patent/JPH06138110A/en not_active Withdrawn
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010514674A (en) * | 2006-10-12 | 2010-05-06 | ベラス ヘルス （インターナショナル） リミテッド | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
| US8748656B2 (en) | 2006-10-12 | 2014-06-10 | Bhi Limited Partnership | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
| JP2015007047A (en) * | 2006-10-12 | 2015-01-15 | ビーエイチアイ リミテッド パートナーシップ | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
| US9499480B2 (en) | 2006-10-12 | 2016-11-22 | Bhi Limited Partnership | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
| US10238611B2 (en) | 2006-10-12 | 2019-03-26 | Bellus Health Inc. | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
| US10857109B2 (en) | 2006-10-12 | 2020-12-08 | Bellus Health, Inc. | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
| US11020360B2 (en) | 2006-10-12 | 2021-06-01 | Bellus Health Inc. | Methods, compounds, compositions and vehicles for delivering 3-amino-1-propanesulfonic acid |
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