JPH0317050A - Device of hydrolyzing protein or peptide - Google Patents
Device of hydrolyzing protein or peptideInfo
- Publication number
- JPH0317050A JPH0317050A JP15080489A JP15080489A JPH0317050A JP H0317050 A JPH0317050 A JP H0317050A JP 15080489 A JP15080489 A JP 15080489A JP 15080489 A JP15080489 A JP 15080489A JP H0317050 A JPH0317050 A JP H0317050A
- Authority
- JP
- Japan
- Prior art keywords
- sample
- specimen
- peptide
- hydrolysis
- protein
- 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 21
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 21
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 18
- 230000003301 hydrolyzing effect Effects 0.000 title claims abstract description 8
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 31
- 150000001413 amino acids Chemical class 0.000 claims abstract description 20
- 239000002253 acid Substances 0.000 claims abstract description 18
- 239000011521 glass Substances 0.000 claims abstract description 14
- 239000000203 mixture Substances 0.000 claims abstract description 7
- 238000010438 heat treatment Methods 0.000 claims abstract description 5
- 239000007787 solid Substances 0.000 claims abstract 2
- 230000032258 transport Effects 0.000 claims abstract 2
- 239000000523 sample Substances 0.000 claims description 82
- 230000007062 hydrolysis Effects 0.000 claims description 26
- 230000007246 mechanism Effects 0.000 claims description 16
- 238000000605 extraction Methods 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 10
- 230000007723 transport mechanism Effects 0.000 claims description 10
- 239000012488 sample solution Substances 0.000 claims description 9
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- 238000010574 gas phase reaction Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims description 2
- 239000011261 inert gas Substances 0.000 claims description 2
- 230000008018 melting Effects 0.000 claims description 2
- 238000002844 melting Methods 0.000 claims description 2
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims 2
- 229910052786 argon Inorganic materials 0.000 claims 1
- 239000000969 carrier Substances 0.000 claims 1
- 238000000354 decomposition reaction Methods 0.000 claims 1
- 230000036571 hydration Effects 0.000 claims 1
- 238000006703 hydration reaction Methods 0.000 claims 1
- 229910052757 nitrogen Inorganic materials 0.000 claims 1
- 238000003825 pressing Methods 0.000 claims 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 abstract description 11
- 238000006243 chemical reaction Methods 0.000 abstract description 6
- 238000011109 contamination Methods 0.000 abstract description 5
- 239000000284 extract Substances 0.000 abstract description 2
- 239000007791 liquid phase Substances 0.000 abstract description 2
- 239000012808 vapor phase Substances 0.000 abstract 1
- 238000000034 method Methods 0.000 description 13
- 230000008016 vaporization Effects 0.000 description 9
- 238000009834 vaporization Methods 0.000 description 7
- 238000010586 diagram Methods 0.000 description 4
- 239000011810 insulating material Substances 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 239000007790 solid phase Substances 0.000 description 4
- 239000007789 gas Substances 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 239000004809 Teflon Substances 0.000 description 2
- 229920006362 Teflon® Polymers 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 239000003708 ampul Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 125000006850 spacer group Chemical group 0.000 description 2
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000005297 pyrex Substances 0.000 description 1
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (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チドのアミノ酸組威
分析の第一段階であるタンパク譬あるいはペプチドの加
水分解を行う装置に関する。DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an apparatus for hydrolyzing proteins or peptides, which is the first step in analyzing the amino acid composition of proteins or peptides.
従来、ア逅ノ酸組戒分析は例えば次の様に行われている
。Conventionally, the analysis of amino acids has been carried out as follows, for example.
（Ｓ．ムーア．Ｈ．スタイン メソッズ インエンザイ
モロジ−（Ｓ．Ｐ．コロビック．Ｎ．Ｏ．カブラ編）
１９６３年 ６＠ ８１９〜８３１ページ，アカデミ
ンクプレス．ニューヨーク）
まず試験管底の乾燥試料に蒸留した共沸点塩酸を加え、
この試験管を氷冷しながら減圧下で封管する．次いで、
このアンプルを１０５℃〜１）０℃で２４時間〜１４４
時間加熱する．次にこのアンプルを開督し、塩酸を蒸発
除去する．最後に加水分解された試料を溶解しア壽ノ酸
分析計に注入する．ところが、最近次田らによって開発
された高速気相加水分解法（次田晧ら、ジャーナル オ
プ バイオケ多ストリー、１９８７年１０２巻１５９３
〜１５９７ページ）は、＆Ｉｌｔｃ分析の第一段階であ
る加水分解を次のように改良している．■加水分解速度
を上げるため反応温度を１５８℃とし、２２．５分間〜
４５分間で迅速に加水分解を完了できるようにした．■
揮発性かつ強酸性の有機酸であるトリフルオロ酢酸を加
えて、疎水性タンパク質の加水分解率を向上させた．■
酸混合蒸気をタンパク質試料に作用させることを利用し
た気相法により、酸自身に含まれるアξノ酸の混入を防
いで正確なア藁ノａｍ＊分析を可能とし、合わせて加水
分解後に残存する酸の除去を簡便にした。(S. Moore. H. Stein Methods in Enzymology (edited by S. P. Corovic and N. O. Kabra)
1963, 6 @ pages 819-831, Akademinck Press. New York) First, add distilled azeotropic hydrochloric acid to the dry sample at the bottom of the test tube.
Seal the test tube under reduced pressure while cooling it on ice. Then,
This ampoule is heated at 105℃~1)0℃ for 24 hours~144℃.
Heat for an hour. Next, open the ampoule and remove the hydrochloric acid by evaporation. Finally, dissolve the hydrolyzed sample and inject it into the amino acid analyzer. However, recently, a high-speed gas phase hydrolysis method developed by Tsugita et al.
(pages 1597 to 1597) improve the hydrolysis, which is the first step in &Iltc analysis, as follows. ■In order to increase the hydrolysis rate, the reaction temperature was set to 158℃, and for 22.5 minutes ~
Hydrolysis can be completed quickly in 45 minutes. ■
Trifluoroacetic acid, a volatile and strongly acidic organic acid, was added to improve the hydrolysis rate of hydrophobic proteins. ■
By using a gas phase method that utilizes acid mixed vapor to act on a protein sample, it is possible to prevent the contamination of amino acids contained in the acid itself, making accurate amino acid analysis possible. This simplifies the removal of acid.
従来の蒸留した共沸点塩酸を用いる加水分解法は２４時
間〜１４４時間という長時間を必要とし、かつ加水分解
後の煩雑な蒸発操作による酸の除去が必要であった．ま
た、液相法であるため酸からの汚染があり、正確なｋＩ
Ｉ威分析は困難であった．特に試料の微量化が要求され
る場合にこの影響は顕著である．また、次田らの開発し
た気相法においても、熟練した技術を要するガラス組工
が必要であり、個人差が避けられず、更に人が試料の取
り扱いの各操作に従事するための汚染が存在することが
欠点として残されている。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、乾燥されたタンパク質試
料の加水分解から、加水分解物のアミノ酸分析計への注
入までの各操作を一切の手操作を用いることなく連続的
に行うこと、および多数の試料を連続的に処理すること
を目的としている．
〔課題を解決するための手段〕
本発明は前記の欠点を除去するためになされたものであ
り、タンパク質あるいはペプチドの試料を担持するガラ
スフィルタの外縁部を溶融固化した試料担体と、タンパ
ク質あるいはペプチドの乾燥された試料を担持した試料
担体の搬送機構と、乾燥された試料へ加圧・加熱下で酸
混合蒸気を供給して固相一気相反応により加水分解反応
を行わせる機構と、加水分解を受けた試料を担持した試
料担体の搬送機構と、前記試料担体からの加水分解され
た試料の抽出機構と、抽出された試料溶液のアえノ酸分
析装置の試料溶液注入装置への送液機構と、前記加水分
解・抽出・送液に用いる流体の供給機構とを備えている
構威とした．〔作用〕
上記のように構威された加水分解装置は試料担体に担持
されたタンパク賞あるいはペプチドのア壽ノ酸＆ｌｌ戒
分析における、搬送、固相一気相反応による加水分解、
抽出およびアミノ酸分析計への注入までをオンライン化
して、各操作を一切の手操作を用いることなく、本質的
な汚染の低下と個人差の排除により正確なアミンａ組威
分析を可能とし、合わせて省力化を図ったものである．
また、一連の操作の連続処理のみならず、多数試料の連
続処理をも行うことができる。The conventional hydrolysis method using distilled azeotropic hydrochloric acid requires a long time of 24 to 144 hours, and requires a complicated evaporation operation to remove the acid after hydrolysis. In addition, since it is a liquid phase method, there is contamination from acids, and accurate kI cannot be obtained.
I-power analysis was difficult. This effect is particularly noticeable when miniaturization of the sample is required. Furthermore, the gas-phase method developed by Tsugita et al. requires a skilled glass worker, individual differences are unavoidable, and contamination occurs due to the human being involved in each operation of sample handling. Its existence remains a drawback. Therefore, in order to solve these conventional drawbacks, this invention allows each operation from hydrolysis of a dried protein sample to injection of the hydrolyzate into an amino acid analyzer to be performed continuously without any manual operations. The purpose is to perform continuous processing and to process a large number of samples continuously. [Means for Solving the Problems] The present invention has been made to eliminate the above-mentioned drawbacks, and includes a sample carrier in which the outer edge of a glass filter supporting a protein or peptide sample is melted and solidified, and a sample carrier that supports a protein or peptide sample. a transport mechanism for a sample carrier supporting a dried sample; a mechanism for supplying acid mixed vapor to the dried sample under pressure and heat to carry out a hydrolysis reaction by a solid phase gas phase reaction; a transport mechanism for a sample carrier carrying a sample that has undergone a reaction, a mechanism for extracting a hydrolyzed sample from the sample carrier, and a transport mechanism for transporting the extracted sample solution to the sample solution injection device of the aenoic acid analyzer. The structure is equipped with a mechanism and a fluid supply mechanism used for the hydrolysis, extraction, and liquid delivery. [Function] The hydrolysis device configured as described above is capable of transporting, hydrolyzing by solid-phase gas-phase reaction,
By making extraction and injection into the amino acid analyzer online, each operation does not require any manual operations, reducing essential contamination and eliminating individual differences, making it possible to perform accurate amine a composition analysis. This is designed to save labor.
Furthermore, it is possible to perform not only continuous processing of a series of operations but also continuous processing of a large number of samples.
以下この発明の実施例を図面に基づいて説明する．
第１図は本発明装置の構威を示すブロック図である．本
発明のタンパク質あるいはペプチドを加水分解する装置
は、乾燥されたタンパク質あるいはペプチドの試料を担
持した試料担体の搬送機構１と、乾燥された試料と酸混
合蒸気との加水分解を行わせる反応機構２と、加水分解
された試料を担持した試料担体の搬送機構３と、加水分
解された試料を試料担体から抽出し、抽出された試料を
アξノ酸分析計に送液する抽出・送液機構４と、反応機
構２と抽出・送液機構４への溶液供給ｍ構５とから構威
されている．タンパク質あるいはペプチドの試料は、１
から４までの各機構を経てアミノ酸へ加水分解され、直
接アξノ酸分析計へ注入される．
次に、本発明のタンパク質あるいはペプチドを加水分解
する装置の動作について述べる．第２図は、本発明に用
いる試料ホルダーの一実施例を示す断面図である．第３
図は、本発明の動作を説明するための工程図である。試
料担体としての試料ホルダー８は、ガラスフィルタの外
縁部を溶融固化したものであり、ガラス７内の内部にガ
ラスフィルタ６が残存している構造となっている．
まず始めに試料溶液の乾燥を行う．試料ホルダー８のガ
ラスフィルタ６にマイクロシリンジ９を用いる試料溶液
を含浸させる，次にこの試料ホルダー８は減圧ボックス
１０に格納される．この減圧ボックス１０は、タイマ１
ｌを備えたｔ磁弁１２を経て真空ボンプ１３と接続して
おり、電磁弁１２の開閉を設定したタイムプログラムに
従い自動的に断続的な吸引を行わせることにより、試料
溶液中の溶媒を除去し試料を乾燥させ、乾燥させた試料
が試料ホルダー８のガラスフィルタ６に担持される。Examples of the present invention will be described below based on the drawings. Figure 1 is a block diagram showing the structure of the device of the present invention. The apparatus for hydrolyzing proteins or peptides of the present invention includes a transport mechanism 1 for a sample carrier carrying a dried protein or peptide sample, and a reaction mechanism 2 for hydrolyzing the dried sample and acid mixed vapor. , a transport mechanism 3 for a sample carrier carrying a hydrolyzed sample, and an extraction/liquid feeding mechanism that extracts the hydrolyzed sample from the sample carrier and sends the extracted sample to the amino acid analyzer. 4, and a solution supply mechanism 5 to the reaction mechanism 2 and the extraction/liquid feeding mechanism 4. A sample of protein or peptide is 1
It is hydrolyzed into amino acids through various mechanisms from 4 to 4, and is directly injected into the amino acid analyzer. Next, the operation of the apparatus for hydrolyzing proteins or peptides of the present invention will be described. FIG. 2 is a sectional view showing one embodiment of a sample holder used in the present invention. Third
The figure is a process diagram for explaining the operation of the present invention. The sample holder 8 as a sample carrier is made by melting and solidifying the outer edge of a glass filter, and has a structure in which the glass filter 6 remains inside the glass 7. First, dry the sample solution. The glass filter 6 of the sample holder 8 is impregnated with a sample solution using a microsyringe 9, and then the sample holder 8 is stored in a vacuum box 10. This decompression box 10 has a timer 1
The solvent in the sample solution is removed by automatically performing intermittent suction according to the time program set to open and close the solenoid valve 12. The sample is then dried, and the dried sample is supported on the glass filter 6 of the sample holder 8.
次に試料ホルダー８は加水分解反応を行わせるステージ
に搬送される．ここで試料ホルダー８はヒータ１４内の
気化チャンバー１５とガラスチューブ１６に固定される
。不活性ガスで酸素をパージした酸混合溶液１７は三方
コソクｌ８を通ってシリンジボンプ１９に吸引され、次
いで三方コンク１８を切り換え、気化チャンバー１５へ
酸混合溶液を供給する．この時、気化チャンバー１５と
試料ホルダー８はヒータ１４で加熱されており、気化チ
ャンバー１５内で蒸気となった酸は試料ホルダー８内を
含む空間に満たされる。この時、ｉｕｉ弁２０は閉して
いるが、この電磁弁２０は圧力計２１と連動させ、試料
ホルダーａ内の圧力を一定に保つように開閉させること
もできる。ここで、試料担体に担持された試料は、酸混
合蒸気との固相一気相反応により加水分解される．次に
、試料ホルダー８内に水蒸気を供給して残存する酸混合
蒸気を除去する．電磁弁２０を通過した酸混合蒸気は、
トラソブ２２で捕集される。Next, the sample holder 8 is transported to a stage where a hydrolysis reaction is performed. Here, the sample holder 8 is fixed to a vaporization chamber 15 and a glass tube 16 within the heater 14. The acid mixed solution 17 purged of oxygen with an inert gas is sucked into the syringe pump 19 through the three-way condenser 18, and then the three-way condenser 18 is switched to supply the acid mixed solution to the vaporization chamber 15. At this time, the vaporization chamber 15 and the sample holder 8 are heated by the heater 14, and the acid vaporized in the vaporization chamber 15 fills the space including the sample holder 8. At this time, the IUI valve 20 is closed, but the solenoid valve 20 can also be opened and closed in conjunction with the pressure gauge 21 to keep the pressure inside the sample holder a constant. Here, the sample supported on the sample carrier is hydrolyzed by a solid-phase gas-phase reaction with the acid mixed vapor. Next, water vapor is supplied into the sample holder 8 to remove the remaining acid mixed vapor. The acid mixed vapor that has passed through the solenoid valve 20 is
Collected by TRASOB 22.
この時、電磁弁２０は開放のままである。At this time, the solenoid valve 20 remains open.
次に試料ホルダー８は、抽出・送液を行うステ一ジに搬
送される．ここで試料ホルダー８はノズル２３により固
定される．希塩酸２４は三方コック２５を経てシリンジ
２６に吸引されている．そして三方コック２５を切り換
え、希塩酸２４により試料を抽出する。試料を溶解した
液体はアミノ酸分析計の注入装置である大方バルブ２７
内のサンプルコイル２８へと送られる。以上の操作に続
くアミノ酸分析については周知であるのでここでは述べ
ない．本実施例における諸条件を第一表にまとめる．第
１表
ここで、加水分解ステージについて、具体的に説明する
．第４図（ａｌは、本発明の加水分解ステージの試料ホ
ルダー解放時の一実施例を示す外観図、第４図ｆｂｌは
、本発明の加水分解ステージの試料ホルダー固定時の一
実施例を示す断面図、第４図（Ｃは、本発明の加水分解
ステージの気化チャンハーの一実施例を示す断面図であ
る。Next, the sample holder 8 is transported to a stage where extraction and liquid delivery are performed. Here, the sample holder 8 is fixed by the nozzle 23. Dilute hydrochloric acid 24 is sucked into a syringe 26 via a three-way cock 25. Then, the three-way cock 25 is switched, and the sample is extracted with dilute hydrochloric acid 24. The liquid in which the sample was dissolved is sent to the main valve 27, which is the injection device of the amino acid analyzer.
The sample coil 28 is sent to the sample coil 28 inside. Amino acid analysis following the above procedure is well known and will not be described here. The various conditions in this example are summarized in Table 1. Table 1 Here, the hydrolysis stage will be specifically explained. Figure 4 (al is an external view of an embodiment of the hydrolysis stage of the present invention when the sample holder is released, Figure 4fbl is an embodiment of the hydrolysis stage of the present invention when the sample holder is fixed) Cross-sectional view, FIG. 4 (C is a cross-sectional view showing one embodiment of the vaporizing chamber of the hydrolysis stage of the present invention.
第３図に示すヒータ１４は気化チャンハ−１５とラバー
ヒーター３０の断熱材３１とカバー３２とからなる固定
ヒーティングユニソト３３と、ガラスチューブｌ６とラ
バーヒーター３０と断熱材３１とカハー３２とエアシリ
ンダー３５とローラー３６とからなる可動ヒーティング
ユニノト３７と、ラハーヒーター３０と断熱材３１とカ
バー３２とローラー３６とからなる支持ユニソト３８と
で構戊されている。これらのユニノトはベース３９上に
図のように配置されている。支持ユニソト３８はローラ
ー３６を備えた台車４０にヒンジ４１を介して乗ってい
る可動ヒーティングユニノト３７と、台車４０は、ロー
ラー３６を用いてレール４２に載せられており、エアシ
リンダー３５を作動させることによって、試料ホルダー
８の固定（第４図（ｂ））と解放（第４図（ａ））を行
うことができる。解放時の支持ユニット４１の戻る位置
決めのため台車４０を貫通するサポートパー４３とスプ
リング４４を設置してある．気化チャンバー１５は、技
付きパイレックスガラスチューブ４５がナット４７と袋
ネジ４８を用いて、フランジを設けたテフロンチューブ
４６と接続されて構戒されている．このテフロンチュー
ブ４６を通して酸混合溶液ｌ９が気化チャンバー１５へ
供給される．
第５図は本発明の抽出送液ステージの構戒の一実施例を
示す外観図である．図は支持台５０上の試料ホルダー８
がエアシリンダー５ｌの作用により、可動ノズル支持プ
レート５２と固定ノズル支持プレート５３とにそれぞれ
挿入された一対のノズル２３により固定された状態を示
している．支持台５０はヒンジ５４を介してスベーサ−
５５と連結されている．固定ノズル支持プレート５３と
スベーサ−５５はべ−ス５６上に図のように固定されて
いる．固定ノズル支持プレート５３には２本のガイドバ
ー５７が貫通しており、そのガイドバー５７の両端でリ
リースプレート５８が支持されている．試料ホルダー８
側のリリースプレート５８と固定ノズル支持プレート５
３との間のガイドパー５７にはスプリング５９が設置し
てあり、試料ホルダー８解放時の駆動源となっている。The heater 14 shown in FIG. 3 includes a fixed heating unit 33 consisting of a vaporizing chamber 15, a heat insulating material 31 of a rubber heater 30, and a cover 32, a glass tube 16, a rubber heater 30, a heat insulating material 31, a heat insulating material 32, and an air heater 33. It consists of a movable heating unit 37 consisting of a cylinder 35 and a roller 36, and a support unit 38 consisting of a Raha heater 30, a heat insulating material 31, a cover 32 and a roller 36. These Uninotes are arranged on the base 39 as shown in the figure. The supporting unit 38 is mounted on a truck 40 equipped with rollers 36 via a hinge 41. The movable heating unit 37 and the truck 40 are placed on a rail 42 using rollers 36, and actuate the air cylinder 35. By doing so, the sample holder 8 can be fixed (FIG. 4(b)) and released (FIG. 4(a)). A support par 43 and a spring 44 are installed that pass through the carriage 40 to determine the return position of the support unit 41 when released. The vaporization chamber 15 is constructed by connecting a fitted Pyrex glass tube 45 to a Teflon tube 46 provided with a flange using nuts 47 and cap screws 48. The acid mixed solution 19 is supplied to the vaporization chamber 15 through this Teflon tube 46. FIG. 5 is an external view showing one embodiment of the arrangement of the extraction liquid feeding stage of the present invention. The figure shows a sample holder 8 on a support stand 50.
is shown fixed by a pair of nozzles 23 inserted into a movable nozzle support plate 52 and a fixed nozzle support plate 53, respectively, due to the action of the air cylinder 5l. The support base 50 is attached to the base via a hinge 54.
It is connected to 55. A fixed nozzle support plate 53 and a baser 55 are fixed on a base 56 as shown in the figure. Two guide bars 57 pass through the fixed nozzle support plate 53, and a release plate 58 is supported at both ends of the guide bars 57. Sample holder 8
Side release plate 58 and fixed nozzle support plate 5
A spring 59 is installed on the guide par 57 between the sample holder 3 and the sample holder 8, and serves as a driving source when the sample holder 8 is released.
第６図（ａｌ〜ｔｄ）は、本発明の試料ホルダーの搬送
機構の動作を説明するための断面図である。FIG. 6 (al to td) is a cross-sectional view for explaining the operation of the transport mechanism of the sample holder of the present invention.
本発明装置の一例としてトレー６０とリテーナ６ｌを介
してトレー６０に固定されたエアシリンダー６２とから
構成されるオートホルダーローダー６３と、加水分解ス
テージ２９と、抽出送液ステージ４９と、格納ケース６
４とが、この順に並んで配置されており、溶媒が除去さ
れ、乾燥された試料を担持する試料ホルダー８がオート
ホルダーローター６３のトレー６０上に搭載されている
（第６図（ａｌ）．次に、試料ホルダー８がエアシリン
ダー６２の作用により、トレー６０から加水分解ステー
ジ２９に搬送される（第６図（ｂｌ），
加水分解ステージ２９で加水分解工程を経た試料ホルダ
ー８は、リテーナ６５を介して台車４０に固定されたエ
アシリンダー６６の作用により、ヒンジ４１によって連
結された支持ユニント３８の一端を上昇せしめ、抽出送
液ステージ４９の支持台５０に搬送される（第６図（Ｃ
ｌ），
抽出送液工程を経た試料ホルダー８は、リテーナ６７を
介してスペーサ５５及びベース５６に固定されたエアシ
リンダー６８の作用により、スベーサ５５にヒンジ５４
を介して連結されている支持台５０の一端が上昇し、格
納ケース６４に格納される（第６回！ｄ＋）．
この様に、試料担体である試料ホルダー８の自動搬送が
達戒され、タンパク質あるいはペプチドのアミノ酸組或
分析における、固相一気相反応による加水分解・抽出及
びア旦ノ酸分析計への注入までを一切の手操作を用いる
ことなく、連続的に行うことができる．
試料保持部としては、試料溶液及び加水分解・抽出に用
いる塩酸等の溶液と反応を行わず、かつ、それらの溶液
を保持する構造が必要であり、ガラスフィルタが用いら
れている。An example of the device of the present invention includes an autoholder loader 63 consisting of a tray 60 and an air cylinder 62 fixed to the tray 60 via a retainer 6l, a hydrolysis stage 29, an extraction liquid feeding stage 49, and a storage case 6.
4 are arranged side by side in this order, and a sample holder 8 carrying a dried sample from which the solvent has been removed is mounted on the tray 60 of the autoholder rotor 63 (FIG. 6(al). Next, the sample holder 8 is transported from the tray 60 to the hydrolysis stage 29 by the action of the air cylinder 62 (Fig. 6 (bl), the sample holder 8 that has undergone the hydrolysis process in the hydrolysis stage 29 is transferred to the retainer 65. By the action of an air cylinder 66 fixed to the trolley 40 through
l) After the extraction liquid feeding process, the sample holder 8 is attached to the spacer 55 by the hinge 54 by the action of the air cylinder 68 fixed to the spacer 55 and the base 56 via the retainer 67.
One end of the support stand 50 connected via the is raised and stored in the storage case 64 (6th! d+). In this way, automatic transportation of the sample holder 8, which is a sample carrier, has been achieved, and it can be used to analyze the amino acid composition of proteins or peptides, from hydrolysis and extraction through solid-phase gas phase reactions to injection into the amino acid analyzer. can be performed continuously without any manual operations. The sample holding section needs to have a structure that does not react with the sample solution and solutions such as hydrochloric acid used for hydrolysis and extraction, and that can hold these solutions, and a glass filter is used.
この発明は以上説明したように、気相加水分解法を採用
し、タンパク質あるいはペプチドのア果ノ酸＆［ｌ或分
析における、試料の加水分解とアミノ酸分析計への注入
までの一切の操作を手操作を用いることなく連続処理可
能とし、また多数試料の連続処理を可能としたことによ
って、熟練を要する手操作及び個人差の排除により正確
なアミノ酸組戒分析を可能とし省力化を図ったものであ
る．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Ｉｓ似のものを除去するものでないこ
とは言うものでもない．As explained above, this invention adopts the gas phase hydrolysis method and eliminates all operations from sample hydrolysis to injection into an amino acid analyzer in the analysis of proteins or peptides. By enabling continuous processing without manual operations and by enabling continuous processing of multiple samples, accurate amino acid composition analysis is possible by eliminating manual operations that require skill and individual differences, resulting in labor savings. It is. It should be noted that the terminology and explanations used herein do not exclude anything similar to Is as long as the features of the present invention are maintained.
第ｌ図は本発明装置の構或を示すプロソク図、第２図は
本発明に用いる試料ホルダーの一実施例を示す断面図、
第３図は本発明の動作を説明するための工程図、第４図
（ａ）は本発明の加水分解ステージの試料ホルダー解放
時の一実施例を示す外観図、第４図（ｂｌは本発明の加
水分解ステージの試料ホルダー固定時の一実施例を示す
断面図、第４図（Ｃ）は本発明の加水分解ステージの気
化チャンバ−の一実施例を示す断面図、第５図は本発明
の抽出・送液ステージの構戒の一実施例を示す外観図、
第６図ｆａ）〜＋ｄ＋は本発明の試料ホルダーの搬送機
構の動作を説明するための断面図である．以上FIG. 1 is a schematic diagram showing the structure of the apparatus of the present invention, and FIG. 2 is a sectional view showing an embodiment of the sample holder used in the present invention.
FIG. 3 is a process diagram for explaining the operation of the present invention, FIG. 4(a) is an external view showing an embodiment of the hydrolysis stage of the present invention when the sample holder is released, and FIG. FIG. 4(C) is a cross-sectional view showing an embodiment of the vaporization chamber of the hydrolysis stage of the present invention when the sample holder is fixed, and FIG. An external view showing an example of the structure of the extraction/liquid feeding stage of the invention,
Figures 6 fa) to +d+ are cross-sectional views for explaining the operation of the transport mechanism of the sample holder of the present invention. that's all
Claims (3)
ラスフィルタの外縁部を溶融固化した試料担体と、タン
パク質あるいはペプチドの乾燥された試料を担持した試
料担体の搬送機構と、乾燥された試料へ加圧・加熱下で
酸混合蒸気を供給して固相−気相反応により加水分解反
応を行わせる機構と、加水分解を受けた試料を担持した
試料担体の搬送機構と、前記試料担体からの加水分解さ
れた試料の抽出機構と、抽出された試料溶液のアミノ酸
分析装置の試料溶液注入装置への送液機構と、前記加水
分解・抽出・送液に用いる流体の供給機構とを備えてい
ることを特徴とするタンパク質あるいはペプチドを加水
分解する装置。(1) A sample carrier made by melting and solidifying the outer edge of a glass filter carrying a protein or peptide sample, a transport mechanism for the sample carrier carrying a dried protein or peptide sample, and a mechanism for pressurizing and applying pressure to the dried sample. A mechanism for supplying acid mixed vapor under heating to perform a hydrolysis reaction by a solid phase-gas phase reaction, a transport mechanism for a sample carrier carrying a hydrolyzed sample, and a mechanism for carrying out a hydrolysis reaction from the sample carrier. The present invention is characterized by comprising: a mechanism for extracting a sample, a mechanism for feeding the extracted sample solution to a sample solution injection device of an amino acid analyzer, and a mechanism for supplying a fluid used for the hydrolysis, extraction, and liquid feeding. A device that hydrolyzes proteins or peptides.
除去には、アルゴンあるいは窒素といった不活性ガスあ
るいは水蒸気あるいはこれらの混合物を用いることを特
徴とする請求項１記載のタンパク質あるいはペプチドの
加水分解装置。(2) The protein or peptide hydration according to claim 1, wherein an inert gas such as argon or nitrogen, water vapor, or a mixture thereof is used to remove the acid mixture from the sample that has undergone hydrolysis. Decomposition equipment.
上の試料担体の連続的な搬送を行うことを特徴とする請
求項１記載のタンパク質あるいはペプチドを加水分解す
る装置。(3) The apparatus for hydrolyzing proteins or peptides according to claim 1, wherein the sample carrier transport mechanism continuously transports one or more sample carriers.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15080489A JPH0317050A (en) | 1989-06-13 | 1989-06-13 | Device of hydrolyzing protein or peptide |
| EP19900302842 EP0388224A3 (en) | 1989-03-17 | 1990-03-16 | Method and apparatus for effecting chemical treatment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15080489A JPH0317050A (en) | 1989-06-13 | 1989-06-13 | Device of hydrolyzing protein or peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0317050A true JPH0317050A (en) | 1991-01-25 |
Family
ID=15504792
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP15080489A Pending JPH0317050A (en) | 1989-03-17 | 1989-06-13 | Device of hydrolyzing protein or peptide |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0317050A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR200447545Y1 (en) * | 2007-10-23 | 2010-02-03 | (주)한보이앤씨 | Accessory for portable telephone with a magnifying glass |
-
1989
- 1989-06-13 JP JP15080489A patent/JPH0317050A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR200447545Y1 (en) * | 2007-10-23 | 2010-02-03 | (주)한보이앤씨 | Accessory for portable telephone with a magnifying glass |
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