JPH01209366A - Separation of amino acid - Google Patents
Separation of amino acidInfo
- Publication number
- JPH01209366A JPH01209366A JP63032691A JP3269188A JPH01209366A JP H01209366 A JPH01209366 A JP H01209366A JP 63032691 A JP63032691 A JP 63032691A JP 3269188 A JP3269188 A JP 3269188A JP H01209366 A JPH01209366 A JP H01209366A
- Authority
- JP
- Japan
- Prior art keywords
- column
- separation
- acetonitrile
- eluent
- amino acid
- 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.)
- Pending
Links
- 238000000926 separation method Methods 0.000 title claims abstract description 18
- 150000001413 amino acids Chemical class 0.000 title claims abstract description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000003480 eluent Substances 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 11
- 239000007853 buffer solution Substances 0.000 claims description 5
- 238000004255 ion exchange chromatography Methods 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims 1
- 239000007924 injection Substances 0.000 claims 1
- 238000010828 elution Methods 0.000 abstract description 6
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract description 5
- 238000004458 analytical method Methods 0.000 abstract description 3
- 239000000872 buffer Substances 0.000 abstract description 3
- 239000003456 ion exchange resin Substances 0.000 abstract description 3
- 229920003303 ion-exchange polymer Polymers 0.000 abstract description 3
- 239000003729 cation exchange resin Substances 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 abstract description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 abstract 3
- 150000001450 anions Chemical class 0.000 abstract 1
- 238000004587 chromatography analysis Methods 0.000 abstract 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 18
- 235000001014 amino acid Nutrition 0.000 description 8
- 235000019441 ethanol Nutrition 0.000 description 8
- 230000000694 effects Effects 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 2
- 239000004475 Arginine Substances 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 2
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 2
- 239000004473 Threonine Substances 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000002522 swelling effect Effects 0.000 description 1
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明はアミノ酸の分離方法に係り、特にアミノ酸のイ
オン交換クロマトグラフ装置における圧力変動が少なく
、分離性の優れたアミノ酸の分離に好適な溶離液を用い
たアミノ酸の分離方法に関するものである。[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a method for separating amino acids, and in particular to an elution method suitable for separating amino acids with small pressure fluctuations and excellent separation properties in an ion exchange chromatography device for amino acids. This invention relates to a method for separating amino acids using a liquid.
従来の溶離液は、クエン酸緩衝液が用いられているが、
これのみでは、トレオニン及びセリンの分離が悪く、そ
の改善法として、緩衝液にエチルアルコールを添加して
用いている。イオン交換法で用いる緩衝液は、最低３種
の緩衝液が用いられ、トレオニン及びセリンは初期に溶
出する成分であるため、エチルアルコールは第１緩衝液
に添加され、その量は約１０〜１６％（ｖ／ｖ）の範囲
にある。The conventional eluent is citrate buffer, but
With this alone, separation of threonine and serine is poor, and as an improvement method, ethyl alcohol is added to the buffer solution. At least three types of buffers are used in the ion exchange method, and since threonine and serine are components that elute initially, ethyl alcohol is added to the first buffer, and the amount is about 10 to 16 % (v/v).
分離能向上、分析時間の短縮をはかろうとするにしたが
い、用いるイオン交換樹脂の粒径を小さくして交率を高
めることが試みられている。このとき、特に問題となる
のはカラム抵抗、すなわち、カラムに緩衝液を流すとき
の圧力上昇である。現在使用しているイオン交換体は、
スチレン／ジビニルベンゼンの共重合体を母体としたも
のがよく、その架橋度は、分難性の面で低いところに抑
える必要がある。 ゛
このため、緩衝液にエチルアルコールを混入したとき、
その膨潤性からカラムの圧力抵抗が上昇する。さらに、
このカラムに流す溶離液は一定のものでなく、グラジェ
ントあるいはステップワイズ法を用いるため、カラム圧
力は常に変動し、充てん剤は徐々に出口側へと詰ってい
き、分離性が低下していく。すなわち、分離カラムの寿
命が短かくなる。In an effort to improve separation performance and shorten analysis time, attempts have been made to reduce the particle size of the ion exchange resin used to increase the cross-over ratio. At this time, a particular problem is column resistance, that is, the pressure increase when the buffer solution flows through the column. The ion exchangers currently in use are
It is preferable to use a styrene/divinylbenzene copolymer as the base material, and the degree of crosslinking must be kept to a low level from the viewpoint of separation resistance.゛For this reason, when ethyl alcohol is mixed into the buffer solution,
Its swelling property increases the pressure resistance of the column. moreover,
The eluent flowing through this column is not constant, but uses a gradient or stepwise method, so the column pressure constantly fluctuates, and the packing material gradually becomes clogged toward the outlet, reducing separation. . In other words, the life of the separation column is shortened.
本発明の目的は、圧力上昇を小さく抑えることができ、
同一の分離性を得ることができるアミノ酸の分離方法を
提供することにある。The purpose of the present invention is to be able to suppress pressure rise to a small level,
The object of the present invention is to provide a method for separating amino acids that can obtain the same separation performance.
上記目的は、溶離液である緩衝液にアセトニトリルを添
加した溶離液を用いてアミノ酸を分離するようにして達
成するようにした。The above object was achieved by separating amino acids using an eluent in which acetonitrile was added to a buffer solution.
アセトニトリルはエチルアルコールに比べ粘性抵抗が小
さく、拡散が速やかであることから、分析成分のピーク
もシャープになり、分離性が向上すると同時に、カラム
圧力低下を満たしてくれる。Since acetonitrile has lower viscous resistance than ethyl alcohol and diffuses more quickly, the peaks of analytical components become sharper, improving separation and at the same time satisfying the drop in column pressure.
その結果、同一の分離性を得るために添加する量も従来
のエチルアルコール量に対し約２／３以下で同じ効果を
得ることができるため、樹脂の膨潤量も小さく抑えるこ
とができる。As a result, the amount of ethyl alcohol added in order to obtain the same separation performance can be about 2/3 or less of the conventional amount of ethyl alcohol to achieve the same effect, and the amount of swelling of the resin can also be kept small.
以下本発明の方法の一実施例を第１図、第２図を用いて
詳細に説明する。An embodiment of the method of the present invention will be described in detail below with reference to FIGS. 1 and 2.
第１表はアミノ酸分析用第１溶離液組成表で、第１表の
Ａは従来用いているエタノールを添加した溶離液で、Ｂ
はエタノールの代りにアセトニトリルを添加した溶離液
である。この二者の溶離液を用いて得られたクロマトグ
ラムがそれぞれ第１図の（Ａ）、（Ｂ）である。有機溶
媒が混入した溶離液では、グリシンとアラニンが接近し
て分離不良になるが、アセトニトリル系では、この点も
エタノールより優れている。Table 1 is the composition of the first eluent for amino acid analysis.
is an eluent containing acetonitrile instead of ethanol. Chromatograms obtained using these two eluents are shown in FIG. 1 (A) and (B), respectively. In an eluent containing an organic solvent, glycine and alanine come close to each other, resulting in poor separation, but acetonitrile is also superior to ethanol in this respect.
第 １ 表
なお、このときの他の分離条件は、次の通りである。分
離カラムサイズ；４．６ｍｍ１．Ｄ、Ｘ６０ｍｍＬ、充
でん剤；日立カスタム陽イオン交換樹脂２６２２ＳＣ，
カラム温度：５７℃、溶離液流量；０．４ｍＱ ／ｍ
１ｎｙカラム圧力；１００〜１２０ｋｇ／ａＪである。Table 1 Other separation conditions at this time are as follows. Separation column size: 4.6mm1. D, X60mmL, filler; Hitachi custom cation exchange resin 2622SC,
Column temperature: 57℃, eluent flow rate: 0.4mQ/m
1ny column pressure: 100 to 120 kg/aJ.
第２図はアミノ酸１８成分を分離した例で、第１図はこ
の前段の部分に相当する。また、後段で溶出する成分（
塩基性）に対してアセトニトリルの添加された溶離を用
いると、アンモニア及びアルギニンの溶出が促進される
。Figure 2 shows an example of separation of 18 amino acid components, and Figure 1 corresponds to the first part. In addition, components eluted in the latter stage (
Using elution with acetonitrile added to (basic) facilitates the elution of ammonia and arginine.
以上説明した本発明によれば、イオン交換樹脂の膨潤及
び溶離液の粘性にともなうカラム圧力上昇が約２／３に
抑えられ、その結果、樹脂の疲労が少なくなり、カラム
の寿命が３倍以上に伸びるという効果がある。According to the present invention described above, the increase in column pressure due to swelling of the ion exchange resin and viscosity of the eluent is suppressed to about two-thirds, and as a result, fatigue of the resin is reduced and the life of the column is increased by more than three times. It has the effect of stretching.
また、分離の最終段ｌ；新らたにアセトニトリルを添加
することにより、アンモニア、アルギニンの溶出位置を
特異的に変えることができるという効果がある。Furthermore, in the final stage of separation, newly adding acetonitrile has the effect that the elution positions of ammonia and arginine can be specifically changed.
第１図は本発明の方法による場合と従来法とのクロマト
グラム結果の比較を示す図、第２図は本発明の方法によ
る実際のクロマ１〜グラムの全体を示した図である。FIG. 1 is a diagram showing a comparison of chromatogram results according to the method of the present invention and the conventional method, and FIG. 2 is a diagram showing the entirety of actual chromatograms 1 to gram according to the method of the present invention.
Claims (1)
注入部と、分離カラムと、該分離カラムの温度制御機能
とを備えたアミノ酸のイオン交換クロマトグラフ装置に
おいて、前記溶離液である緩衝液にアセトニトリルを添
加した溶離液を用いてアミノ酸を分離するようにしたこ
とを特徴とするアミノ酸の分離方法。1. In an amino acid ion exchange chromatography apparatus comprising an eluent, a pump for feeding the eluent, a sample injection section, a separation column, and a temperature control function for the separation column, the eluent is A method for separating amino acids, characterized in that amino acids are separated using an eluent prepared by adding acetonitrile to a buffer solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63032691A JPH01209366A (en) | 1988-02-17 | 1988-02-17 | Separation of amino acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63032691A JPH01209366A (en) | 1988-02-17 | 1988-02-17 | Separation of amino acid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01209366A true JPH01209366A (en) | 1989-08-23 |
Family
ID=12365884
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63032691A Pending JPH01209366A (en) | 1988-02-17 | 1988-02-17 | Separation of amino acid |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01209366A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003528602A (en) * | 2000-03-29 | 2003-09-30 | アーカー−ダニエルズ−ミッドランド カンパニー | Method for separating basic amino acids from fermentation broth |
| CN104922932A (en) * | 2015-05-26 | 2015-09-23 | 安徽皖东树脂科技有限公司 | Application method of special resin for extracting amino acid |
| CN109507314A (en) * | 2018-10-29 | 2019-03-22 | 中科谱研（北京）科技有限公司 | The ion chromatography method of sodium, potassium, magnesium, calcium in Amino Acid Compound Injection |
-
1988
- 1988-02-17 JP JP63032691A patent/JPH01209366A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003528602A (en) * | 2000-03-29 | 2003-09-30 | アーカー−ダニエルズ−ミッドランド カンパニー | Method for separating basic amino acids from fermentation broth |
| JP2011177709A (en) * | 2000-03-29 | 2011-09-15 | Archer-Daniels-Midland Co | Method for separating basic amino acid from fermentation broth |
| CN104922932A (en) * | 2015-05-26 | 2015-09-23 | 安徽皖东树脂科技有限公司 | Application method of special resin for extracting amino acid |
| CN109507314A (en) * | 2018-10-29 | 2019-03-22 | 中科谱研（北京）科技有限公司 | The ion chromatography method of sodium, potassium, magnesium, calcium in Amino Acid Compound Injection |
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