JPH01180457A - Hydrolyzing device for protein or peptide - Google Patents
Hydrolyzing device for protein or peptideInfo
- Publication number
- JPH01180457A JPH01180457A JP63005462A JP546288A JPH01180457A JP H01180457 A JPH01180457 A JP H01180457A JP 63005462 A JP63005462 A JP 63005462A JP 546288 A JP546288 A JP 546288A JP H01180457 A JPH01180457 A JP H01180457A
- Authority
- JP
- Japan
- Prior art keywords
- sample
- acid
- protein
- amino acid
- peptide
- 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
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 17
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 17
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 9
- 230000003301 hydrolyzing effect Effects 0.000 title abstract 2
- 239000002253 acid Substances 0.000 claims abstract description 27
- 150000001413 amino acids Chemical class 0.000 claims abstract description 16
- 238000006243 chemical reaction Methods 0.000 claims abstract description 9
- 238000010438 heat treatment Methods 0.000 claims abstract description 9
- 238000006460 hydrolysis reaction Methods 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 12
- 230000007062 hydrolysis Effects 0.000 claims description 9
- 238000002347 injection Methods 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- 239000011259 mixed solution Substances 0.000 claims description 3
- 238000011282 treatment Methods 0.000 claims description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 9
- 239000011347 resin Substances 0.000 abstract description 5
- 229920005989 resin Polymers 0.000 abstract description 5
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 3
- 239000000523 sample Substances 0.000 description 19
- 238000000034 method Methods 0.000 description 8
- 238000010586 diagram Methods 0.000 description 5
- 102000004196 processed proteins & peptides Human genes 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 229910001873 dinitrogen Inorganic materials 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 102000051325 Glucagon Human genes 0.000 description 2
- 108060003199 Glucagon Proteins 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 2
- 229960004666 glucagon Drugs 0.000 description 2
- 239000011261 inert gas Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- -1 Gyflon Polymers 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 Teflon® Polymers 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000000813 peptide hormone Substances 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は、タンパク質あるいはペプタイドを加水分解反
応によりその構成成分であるアミノ酸に分解する装置に
関する。DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an apparatus for decomposing proteins or peptides into their constituent amino acids through a hydrolysis reaction.
本発明は、タンパク質あるいはペプタイドの加水分解反
応における、試料溶液と酸溶液との混合、混合された溶
液の加熱、加熱後の溶液からの酸の除去およびアミノ酸
分析針への試料の注入を自動化し、高速・正確・かつ簡
便にタンパク質のアミノ酸組成値を得ようとするもので
ある。The present invention automates the mixing of a sample solution and an acid solution, the heating of the mixed solution, the removal of the acid from the solution after heating, and the injection of the sample into an amino acid analysis needle in the hydrolysis reaction of proteins or peptides. The aim is to obtain amino acid composition values of proteins quickly, accurately, and easily.
従来、第４図に示すように試料をあらかじめ底に乾燥し
ておいた試験管に共沸塩酸あるいは塩酸と揮発性の有機
酸を入れ、氷冷上真空封管した後１０５℃で２４時間、
あるいは１６６℃で２５分間加熱することによる加水分
解法が知られていた０例えばＳ・ムーアら メソフヅ
イン エンザイモロジ−６巻 ８１９〜８３１ページ
１９６３年発行および、次田晧ら、ヨーロピアン ジャ
ーナルオプ バイオケミストリー １２４巻 ５８５〜
５８８ページ １９８２年発行にこれらの方法が報告さ
れている。Conventionally, as shown in Figure 4, azeotropic hydrochloric acid or hydrochloric acid and a volatile organic acid are placed in a test tube with a sample previously dried at the bottom, the tube is vacuum-sealed on ice, and then heated at 105°C for 24 hours.
Alternatively, a hydrolysis method by heating at 166°C for 25 minutes was known. For example, S. Moore et al.
In Enzymology, Volume 6, Pages 819-831
Published in 1963 and published by Akira Tsugita et al., European Journal of Biochemistry, Vol. 124, 585-
These methods are reported on page 588, published in 1982.
しかし従来の加水分解法は、ガスバーナーの炎と冷却ト
ラップを備えた真空ポンプを用いる減圧下の試験管の封
管、加熱後の開管、開管した試験管内の酸のロータリー
エバポレータによる蒸発除去といった煩雑なマニュアル
操作を含み、更に蒸発乾固された試料を再溶解しなけれ
ばアミノ酸分析に供するサンプルが調整できないという
欠点があった。そこでこの発明は、従来のこのような欠
点を解決するため、装置に導入されたタンパク質あるい
はペプタイドを閉ざされた流路内で加水分解し、その処
理された試料をマニュアル操作を加えることなく自動的
にアミノ酸分析装置に注入して、高速・正確かつ簡便に
タンパク質あるいはペプタイドの７ミノ＠組成分析値を
得ることを目的としている。However, conventional hydrolysis methods involve sealing a test tube under reduced pressure using a gas burner flame and a vacuum pump with a cold trap, opening the tube after heating, and removing the acid in the open test tube by evaporation using a rotary evaporator. This method involves complicated manual operations such as evaporation to dryness, and has the disadvantage that a sample for amino acid analysis cannot be prepared without redissolving the sample that has been evaporated to dryness. Therefore, in order to solve these conventional drawbacks, this invention hydrolyzes proteins or peptides introduced into the device in a closed channel, and automatically processes the processed sample without manual operation. The purpose is to quickly, accurately, and easily obtain 7-mino@compositional analysis values for proteins or peptides by injecting them into an amino acid analyzer.
上記問題点を解決するためにこの発明は、注入装置から
導入された試料と酸溶液との混合、混合された溶液の反
応装置への移送と加熱による加水分解反応の進行、除去
装置を用いた反応溶液からの酸の除去およびこれらの処
理の完了した試料のアミノ酸分析針への導入を閉鎖系内
でシーケンシャルに行わせることとした。In order to solve the above problems, this invention mixes a sample introduced from an injection device with an acid solution, transfers the mixed solution to a reaction device, progresses the hydrolysis reaction by heating, and uses a removal device. The removal of acid from the reaction solution and the introduction of the sample after these treatments into the amino acid analysis needle were performed sequentially in a closed system.
上記のように構成された加水分解装置においては、注入
器から導入された試料の酸溶液との混合、反応装置にお
ける加熱、加水分解、除去装置における反応後の酸の除
去としてアミノ酸分析計への注入が閉ざされた流路内で
連続して行われるのである。In the hydrolysis apparatus configured as described above, the sample introduced from the syringe is mixed with the acid solution, heated and hydrolyzed in the reaction apparatus, and the acid is removed after the reaction in the removal apparatus and sent to the amino acid analyzer. Injection takes place continuously within a closed channel.
以下にこの発明の実施例を図面に基づいて説明する。第
１図のブロック図に示す如く、本発明の加水分解装置は
酸貯蔵槽１．その貯蔵槽に蓄えられた酸を流路内へ供給
する送液装置２．流路内にタンパク賞試料を導入する注
入装置３．酸溶液とタンパク質試料との混合物を加熱し
加水分解反応を進行させる反応装置４および反応混液か
ら酸を除去するための除酸装置５がこの順にチューブに
よって連結されたものである。酸貯蔵槽１はあらかじめ
酸素を除去した酸溶液を蓄えるものであり装置作動、中
の酸素の混入を除く構造となっている。Embodiments of the present invention will be described below based on the drawings. As shown in the block diagram of FIG. 1, the hydrolysis apparatus of the present invention includes an acid storage tank 1. 2. Liquid feeding device that supplies the acid stored in the storage tank into the flow path. Injection device for introducing the protein prize sample into the channel 3. A reaction device 4 for heating a mixture of an acid solution and a protein sample to advance a hydrolysis reaction and an acid removal device 5 for removing acid from the reaction mixture are connected in this order by a tube. The acid storage tank 1 stores an acid solution from which oxygen has been removed in advance, and has a structure that prevents oxygen from entering the tank during device operation.
第２図はここに記した機能を満たす加水分解装置の一例
である。貯蔵容器７に蓄えられているｆａ温溶液窒素ボ
ンベ６から供給される窒素ガスで加圧されている。窒素
ガスの代わりに酸素を含まない、いかなる不活性ガズを
も使用可能である。酸溶液は三方コック８を経てシリン
ジポンプ９に吸引された後に吐出される０本例ではシリ
ンジポンプを用いているが、ペリスタリンクポンプある
いはプランジャーポンプといったいかなる定量送液装置
をも使用可能である。タンパク質試料は六方パルプ１０
から系内に自動的に導入される。試料の導入は手動でも
よく、また他のいかなる注入装置をも本装置に組み込む
ことが可能である。酸溶液と混合された試料は恒温槽１
２内の反応コイル１１に導かれ加水分解される。恒温加
熱には、ブロック恒温槽、オイルバス、赤外線ヒータと
いったいかなる方法も採用可能である。加熱加水分解さ
れた試料は樹脂チューブ１３内に導かれる。このチュー
ブにはテフロン、グイフロン、あるいはシリコンを含む
気体透過性のいかなる樹脂をも使用可能である。FIG. 2 is an example of a hydrolysis device that satisfies the functions described herein. It is pressurized with nitrogen gas supplied from a fa hot solution nitrogen cylinder 6 stored in a storage container 7. Any oxygen-free inert gas can be used in place of nitrogen gas. The acid solution is sucked into a syringe pump 9 via a three-way cock 8 and then discharged. Although a syringe pump is used in this example, any metering liquid delivery device such as a peristaltic link pump or a plunger pump can be used. be. Protein sample is hexagonal pulp 10
automatically introduced into the system. Sample introduction can be done manually, and any other injection device can be incorporated into the device. The sample mixed with the acid solution is placed in a constant temperature bath 1.
It is guided to the reaction coil 11 in 2 and is hydrolyzed. Any method such as a block constant temperature bath, an oil bath, or an infrared heater can be used for constant temperature heating. The heated and hydrolyzed sample is introduced into the resin tube 13. The tube can be made of any gas permeable resin including Teflon, Gyflon, or silicone.
試料中に含まれる酸は樹脂チューブ１３を通過中に真空
ポンプ１４の減圧により系内より除去される。The acid contained in the sample is removed from the system by the reduced pressure of the vacuum pump 14 while passing through the resin tube 13.
系内からの酸の除去には第３図に示したように、気体透
過性を持つ樹脂チューブ１３とそれをおおう外筒１５と
の間に窒素ガス等の不活性ガスを流す方法を用いること
もできる。酸を除かれた試料はアミノ酸分析装置の自動
試料注入装置へ導かれる。To remove the acid from the system, as shown in Figure 3, a method is used in which an inert gas such as nitrogen gas is passed between the gas-permeable resin tube 13 and the outer cylinder 15 that covers it. You can also do it. The sample from which the acid has been removed is guided to the automatic sample injection device of the amino acid analyzer.
第５図は本装置により処理された、ペプチドホルモンの
一種であるグルカゴンのアミノ酸分析のチャートである
。この結果は本装置を用いることにより、処理の途中に
いかなるマニュアル操作を加えることなくタンパク質試
料９アミノ酸組成を得ることが可能であることを示して
いる。FIG. 5 is a chart of amino acid analysis of glucagon, a type of peptide hormone, processed by this apparatus. This result shows that by using this apparatus, it is possible to obtain a 9-amino acid composition of a protein sample without adding any manual operations during the processing.
（発明の効果〕
この発明は以上説明したように、装置に導入されたタン
パク質あるいはペプタイドを閉ざされた流路内で加水分
解し、その処理された試料を自動的にアミノ酸分析装置
に注入して高、速・正確かつ簡便にタンパク質のアミノ
酸組成分析値を得る効果がある。(Effects of the Invention) As explained above, the present invention hydrolyzes proteins or peptides introduced into the device in a closed channel, and automatically injects the treated sample into the amino acid analyzer. It has the effect of obtaining protein amino acid composition analysis values quickly, accurately, and easily.
なお、ここで使用した用語及び説明は本発明の特徴が維
持される限り類似のものを除去するものではないことは
言うまでもない。It goes without saying that the terms and explanations used herein do not exclude similarities as long as the characteristics of the present invention are maintained.
第１図は加水分解装置の構成を示すブロック図、第２図
は加水分解装置の構成図、第３図は除酸装置の一例を示
す構成図、第４図はグルカゴン加水分解物のアミノ酸分
析のチャート図、第５図は従来の加水分解法の説明図で
ある。
一以上
出願人 セイコー電子工業株式会社
；
■
アミノ政丸づｆ第４装置
加水分′ｌ＃伎１の構成を示すブロック図第１図
フ゛ルカゴン加水＋ぬ物のアミノ叙分析のナヤート第
４ 図Figure 1 is a block diagram showing the configuration of a hydrolysis device, Figure 2 is a configuration diagram of a hydrolysis device, Figure 3 is a configuration diagram showing an example of an acid removal device, and Figure 4 is an amino acid analysis of glucagon hydrolyzate. FIG. 5 is an explanatory diagram of the conventional hydrolysis method. One or more applicants: Seiko Electronics Co., Ltd.
4 Figure
Claims (1)
れた溶液の反応装置への移送と加熱による加水分解反応
の進行、除去装置を用いた反応溶液からの酸の除去およ
びこれらの処理の完了した試料のアミノ酸分析計への導
入を、閉鎖系内でシーケンシャルに行わせることを特徴
とする、タンパク質あるいはペプタイドの加水分解装置
。Mixing the sample introduced from the injection device with the acid solution, transferring the mixed solution to the reaction device and proceeding with the hydrolysis reaction by heating, removing the acid from the reaction solution using the removal device, and performing these treatments. A protein or peptide hydrolysis device characterized by sequentially introducing a completed sample into an amino acid analyzer in a closed system.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63005462A JPH01180457A (en) | 1988-01-13 | 1988-01-13 | Hydrolyzing device for protein or peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63005462A JPH01180457A (en) | 1988-01-13 | 1988-01-13 | Hydrolyzing device for protein or peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01180457A true JPH01180457A (en) | 1989-07-18 |
Family
ID=11611895
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63005462A Pending JPH01180457A (en) | 1988-01-13 | 1988-01-13 | Hydrolyzing device for protein or peptide |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01180457A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0377855A (en) * | 1989-08-18 | 1991-04-03 | Seiko Instr Inc | Device for hydrolyzing protein or peptide |
| JPH0377853A (en) * | 1989-08-18 | 1991-04-03 | Seiko Instr Inc | Device for hydrolyzing protein or peptide |
| GB2246719A (en) * | 1990-08-10 | 1992-02-12 | Savant Instr | Hydrolysis unit and method |
| CN102062708A (en) * | 2010-11-26 | 2011-05-18 | 兰州大学 | Device for pretreatment of amino acid test sample |
| CN103063785A (en) * | 2012-12-27 | 2013-04-24 | 大连依利特分析仪器有限公司 | Multi-functional pretreatment device used for amino acid analysis |
-
1988
- 1988-01-13 JP JP63005462A patent/JPH01180457A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0377855A (en) * | 1989-08-18 | 1991-04-03 | Seiko Instr Inc | Device for hydrolyzing protein or peptide |
| JPH0377853A (en) * | 1989-08-18 | 1991-04-03 | Seiko Instr Inc | Device for hydrolyzing protein or peptide |
| GB2246719A (en) * | 1990-08-10 | 1992-02-12 | Savant Instr | Hydrolysis unit and method |
| US5252485A (en) * | 1990-08-10 | 1993-10-12 | Savant Instruments, Inc. | Unit for hydrolyzing amino-acid containing specimens |
| CN102062708A (en) * | 2010-11-26 | 2011-05-18 | 兰州大学 | Device for pretreatment of amino acid test sample |
| CN103063785A (en) * | 2012-12-27 | 2013-04-24 | 大连依利特分析仪器有限公司 | Multi-functional pretreatment device used for amino acid analysis |
| CN103063785B (en) * | 2012-12-27 | 2014-11-12 | 大连依利特分析仪器有限公司 | Multi-functional pretreatment device and method used for amino acid analysis |
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