JPH0317048A - Device of hydrolyzing protein or peptide - Google Patents
Device of hydrolyzing protein or peptideInfo
- Publication number
- JPH0317048A JPH0317048A JP15080189A JP15080189A JPH0317048A JP H0317048 A JPH0317048 A JP H0317048A JP 15080189 A JP15080189 A JP 15080189A JP 15080189 A JP15080189 A JP 15080189A JP H0317048 A JPH0317048 A JP H0317048A
- Authority
- JP
- Japan
- Prior art keywords
- sample
- specimen
- glass
- peptide
- 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 23
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 23
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 20
- 230000003301 hydrolyzing effect Effects 0.000 title claims abstract description 10
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 31
- 150000001413 amino acids Chemical class 0.000 claims abstract description 19
- 239000002253 acid Substances 0.000 claims abstract description 18
- 239000011521 glass Substances 0.000 claims abstract description 17
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 239000005373 porous glass Substances 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 87
- 230000007062 hydrolysis Effects 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 15
- 230000007246 mechanism Effects 0.000 claims description 15
- 238000000605 extraction Methods 0.000 claims description 11
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 8
- 239000012488 sample solution Substances 0.000 claims description 8
- 230000007723 transport mechanism Effects 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 239000000919 ceramic Substances 0.000 claims description 5
- 238000010574 gas phase reaction Methods 0.000 claims description 5
- 239000003365 glass fiber Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000000969 carrier Substances 0.000 claims description 2
- 239000012530 fluid Substances 0.000 claims description 2
- 239000011261 inert gas Substances 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
- 238000000354 decomposition reaction Methods 0.000 claims 1
- 239000008187 granular material Substances 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
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 abstract description 11
- 238000006243 chemical reaction Methods 0.000 abstract description 5
- 238000011109 contamination Methods 0.000 abstract description 5
- 238000010438 heat treatment Methods 0.000 abstract description 4
- 239000000284 extract Substances 0.000 abstract description 3
- 239000007791 liquid phase Substances 0.000 abstract description 2
- 239000012808 vapor phase Substances 0.000 abstract 1
- 238000000034 method Methods 0.000 description 15
- 238000009834 vaporization Methods 0.000 description 9
- 230000008016 vaporization Effects 0.000 description 9
- 238000010586 diagram Methods 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 239000007790 solid phase Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 239000011810 insulating material Substances 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 125000006850 spacer group Chemical group 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
- 230000002378 acidificating effect Effects 0.000 description 2
- 239000003708 ampul Substances 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 1
- 239000011260 aqueous acid Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012546 transfer Methods 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戒
分析の第一段階であるタンパク質あるいはペプチドの加
水分解を行う装置に関する．〔従来の技術〕
従来、アξノ酸組威分析は例えば次の様に行われている
．
（Ｓ．ムーア，Ｈ．スタイン メソフズ インエンザイ
モロジ−（Ｓ．Ｐ．コロビック．Ｎ．Ｏ．カプラ編＞
１９６３年 ６＠ ８１９〜８３１ページ，アカデξ
ツクプレス．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℃〜１１０℃で２４時間〜１４４
時間加熱する．次にこのアンプルを開管し、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ξ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
ブ バイオケミストリー、１９８７年１０２１１ １５
９３〜１５９７ページ）は、＆ｌ戒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ａ酸であるトリフルオロ酢酸を加
えて、疎水性タンパク賞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もノ酸の混入を防
いで正確なアξノ酸組戊分折を可能とし、合わせて加水
分解後に残存する酸の除去を簡便にした．
〔発明が解決しようとする！ｉｌ！）
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、正確な組戒
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、Ｐ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龜行
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。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 amino acid composition analysis of proteins or peptides. [Prior art] Conventionally, amino acid composition analysis has been carried out as follows, for example. (S. Moore, H. Stein Methoff's In Enzymology (S.P. Corobic. N.O. Capra ed.)>
1963 6 @ pages 819-831, Akade ξ
Tsukupress. New York) First, add distilled azeotropic hydrochloric acid to the dry sample at the bottom of the test tube.
The test tube is sealed under reduced pressure while cooling on ice. Then,
This ampoule was heated at 105℃ to 110℃ for 24 hours to 144 hours.
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 ξ-anoic acid analyzer. However, the fast gas phase hydrolysis method recently developed by Tsugita et al. (Aki Tsugita et al., Journal of Biochemistry, 1987, 10211 15
(pp. 93-1597) improves the hydrolysis, which is the first step in &l precept analysis, as follows. ■In order to increase the hydrolysis rate, the reaction temperature was set to 158℃, and for 22.5 minutes ~
Hydrolysis could be completed quickly in 45 minutes. ■
Trifluoroacetic acid, a volatile and strongly acidic aqueous acid, was added to improve the hydrolysis rate of hydrophobic proteins. ■
By using a gas phase method that uses mixed acid vapor to act on a protein sample, it is possible to prevent the contamination of amino acids contained in the acid itself, enabling accurate ano acid composition analysis, and also to perform hydrolysis. This makes it easier to remove the acid that remains afterwards. [Invention tries to solve! Il! ) The conventional hydrolysis method using distilled azeotropic hydrochloric acid requires a long time of 24 to 144 hours, and requires removal of acid by a complicated evaporation operation after hydrolysis. In addition, since it is a liquid phase method, there is contamination from acids, making accurate analysis of the composition difficult. This effect is particularly noticeable when miniaturization of the sample is required. Furthermore, the gas-phase method developed by Tsugita et al. requires glass fabrication that requires sophisticated techniques, and individual differences are unavoidable, and contamination due to the human being engaged in each operation of sample handling is also a problem. The existence of this 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 targeted measurements and to process a large number of samples continuously.
上記のように構戊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ブチドのアξノ酸＆［ｌ或
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ξノ酸分析針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ξノ＃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。The hydrolysis apparatus constructed as described above is capable of hydrolyzing proteins or peptides supported on sample carriers by transportation, solid-phase gas-phase reaction,
By making extraction and injection into the amino acid analysis needle online, each operation does not require any manual operations, reducing essential contamination and eliminating individual differences, resulting in accurate amino acid analysis. This makes it possible to do this, and also saves labor.
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。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ｆＪＩ威を示すブロック図である
。本発明のタンパク賞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横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・送液機溝４と、反
応機構２と抽出・送液機横４への溶液供給機構５とから
構威されている．タンパク質あるいはペブチドの試料は
、１から４までの各機構を経てアミノ酸へ加水分解され
、直接アミノ酸分析計へ注入される。Examples of the present invention will be described below based on the drawings. FIG. 1 is a block diagram showing the fJI performance of the device of the present invention. The apparatus for hydrolyzing protein or peptide of the present invention comprises a transport mechanism 1 for a sample carrier carrying a dried protein or peptide sample, and a reaction machine for hydrolyzing the dried sample and acid mixed vapor. horizontal 2, a transport mechanism 3 for the sample carrier carrying the hydrolyzed sample, and an extraction/transfer mechanism that extracts the hydrolyzed sample from the sample carrier and sends the extracted sample to the amino acid analyzer. It consists of a liquid machine groove 4, a reaction mechanism 2, and a solution supply mechanism 5 to the extraction/liquid feeding machine side 4. A protein or peptide sample is hydrolyzed into amino acids through mechanisms 1 to 4 and directly injected into an amino acid analyzer.
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、支持部としてのガラ
ス外筒６内に試料の保持部としての多孔質ガラスからな
るガラスフィルタ７を挿入し、ガラス外筒６の両端の外
径が中央部の外径より小さくした構造をとっている。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 has a glass filter 7 made of porous glass as a sample holding part inserted into a glass outer cylinder 6 as a support part, and the outer diameter of both ends of the glass outer cylinder 6 is the same as that of the center part. It has a structure that is smaller than the outer diameter.
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７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
ｌＯに格納される。この滅圧ボ，クス１０は、クィマ１
１を備えた電磁弁１２を経て真空ポンプｌ３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
？ｆＩ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
。First, the sample solution is dried. Impregnate the glass filter 7 of the sample holder 8 with the sample solution using the microsyringe 9. Next, this sample holder 8 is stored in a vacuum box IO. This decompression box 10 is
It is connected to a vacuum pump 13 through a solenoid valve 12 equipped with a solenoid valve 12, and by automatically performing intermittent suction according to a time program set to open and close the solenoid valve 12, The solvent in the fI liquid is removed and the sample is 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ｌ７は三方
コンク■８を通ってシリンジポンブ１９に吸引され、次
いで三方コック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。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 cock 18 is switched to supply the acid mixed solution to the vaporization chamber 15.
この時、気化チャンバー１５と試料ホルダー８はヒーク
ｌ４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。At this time, the vaporization chamber 15 and the sample holder 8 are heated by the heat 14, and the acid vaporized in the vaporization chamber 15 fills the space including the sample holder 8.
この時、電磁弁２０は閉じているが、この電磁弁２０は
圧力計２１と連動させ、試料ホルダ８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、酸混合澤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２
０を通過した酸混合蒸気は、トラップ２２で捕集される
。At this time, the solenoid 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 8 constant. Here, the sample supported on the sample carrier is hydrolyzed by a solid-phase one-gas phase reaction with acid-mixed gas. Next, water vapor is supplied into the sample holder 8 to remove the remaining acid mixed vapor. Solenoid valve 2
The acid mixed vapor that has passed through the trap 22 is collected by the trap 22.
この時、ａ磁弁２０は開放のままである。At this time, the a magnetic valve 20 remains open.
次に試料ホルダー８は、抽出・送液を行うステージに搬
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は三方コフク２５を経てシリンジ
２６に吸引されている．そして三方コソク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六方バルブ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ξ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ｆａｌ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拭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 feeding are performed. Here, the sample holder 8 is fixed by the nozzle 23. Dilute hydrochloric acid 24 is aspirated into a syringe 26 via a three-way pipe 25. Then, switch the three-way gasket 25 and extract the sample with dilute hydrochloric acid 24. The liquid in which the sample was dissolved is passed through the six-way valve 27, which is the injection device of the amino acid analyzer.
is sent to the sample coil 28 inside. The 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 fal is an external view showing an example of the hydrolysis stage of the present invention when the sample holder is released, and Figure 4 (bl is an example of the hydrolysis stage of the present invention when the wipe holder is fixed). The cross-sectional view, FIG. 4(c), is a cross-sectional view showing one embodiment of the vaporization chamber of the hydrolysis stage of the present invention.
第３図に示すヒータ１４は気化チャンバー１５とラバー
ヒーター３０の断熱材３ｌとカバー３２とからなる固定
ヒーティングユニット３３と、ガラスチューブｌ６とラ
バーヒーター３０と断熱材３１とカバー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
ユニット３７と、ラバーヒーター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ット３８と
で構威されている。これらのユニットはベース３９上に
図のように配置されている．支持ユニット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ソト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ｔａｌ ）を
行うことができる．解放時の支持ユニット４ｌの戻る位
置決めのため台車４０を貫通するサボ７トバー４３とス
プリング４４を設゛置してある．気化チャンバー１５は
、技付きパイレソクスガラスチューブ４５がナント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と接続されて構成されている．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１
５へ供給される．
第５図は本発明の抽出送液ステージの構威の一実施例を
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により固賂された状態を示
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はベース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
−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との間のガイドバ−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 vaporization chamber 15, a heat insulating material 3l of a rubber heater 30, and a cover 32, a glass tube l6, a rubber heater 30, a heat insulating material 31, a cover 32, and an air cylinder 35. and a movable heating unit 37 consisting of a roller 36 and a support unit 38 consisting of a rubber heater 30, a heat insulating material 31, a cover 32 and a roller 36. These units are arranged on the base 39 as shown in the figure. The support unit 38 is mounted on a movable heating unit 37 which is mounted on a carriage 40 equipped with rollers 36 via a hinge 41, and the carriage 40 is mounted on a rail 42 using rollers 36, and operates the air cylinder 35. By doing so, the sample holder 8 can be fixed (Fig. 4(b)) and released (Fig. 4 (tal)). A sabot 7 bar 43 and a spring 44 passing through the cart 40 are installed to position the support unit 4l back when released. The vaporization chamber 15 has a Pyresox glass tube 45 made of Nantes 47.
and a Teflon tube 46 provided with a flange using cap screws 48. The acid mixed solution 19 is supplied to the vaporization chamber 1 through this Teflon tube 46.
5. FIG. 5 is an external view showing one embodiment of the structure of the extraction liquid feeding stage of the present invention. The figure shows a sample holder 8 on a support stand 50.
The figure shows a state in which the nozzles 23 are fixed by the pair of nozzles 23 inserted into the movable nozzle support plate 52 and the 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 spacer 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 bar 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〜（ｄｌ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。FIGS. 6A to 6D are cross-sectional views 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６１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
ホルダーローダー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Ｑ）））．
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４ｌ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
））．
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図＋ｄ））．
この様に、試料担体である試料ホルダー８の自ｖＪｌＩ
！送が達成され、タンパク質あるいはペブチドのアミノ
１５２組成分析における、固相一気相反応による加水分
解・抽出及びアξノ酸分析計への注入までを１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。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 61, a hydrolysis stage 29, an extraction liquid feeding stage 49, and a storage case 6.
4 are arranged 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 loader 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. 6Q)). The sample holder 8 that has undergone the hydrolysis process in the hydrolysis stage 29 is moved up by the action of an air cylinder 66 fixed to the trolley 40 via a retainer 65 to raise one end of the support unit 38 connected by the hinge 4l, and the sample holder 8 is extracted and transported. The liquid is transported to the support stand 50 of the liquid stage 49 (Fig. 6 (C)
))． The sample holder 8 that has undergone the extraction liquid feeding process 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, which is connected via the support stand 50, rises and is stored in the storage case 64 (Fig. 6+d)). In this way, the self-vJlI of the sample holder 8, which is the sample carrier,
! In the amino-152 composition analysis of proteins or peptides, hydrolysis/extraction by solid-phase gas phase reaction and injection into the amino acid analyzer can be carried out continuously without any manual operations. It can be carried out.
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、ガラス繊維を織ったもの
、ガラスを多孔質に加工したもの、ガラス又はセラミッ
クスのビーズを詰めたものが使用される．
（発明の効果）
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組戒分析に
おける、試料の加水分解とアξ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
旦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
持される限り類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．The sample holding part must 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, such as woven glass fibers or porous glass. These are processed into clay or filled with glass or ceramic beads. (Effects of the Invention) As explained above, this invention adopts the gas phase hydrolysis method, and in the analysis of protein or peptide amino acids, the sample is hydrolyzed and injected into the amino acid analyzer. By making it possible to continuously process all the steps up to this point without using any manual operations, and by making it possible to process a large number of samples continuously, it is possible to perform accurate amino acid composition analysis by eliminating manual operations that require skill and individual differences. This makes it possible to save labor.
It should be noted that the terms and explanations used herein do not exclude similar terms as long as the characteristics of the present invention are maintained.
第１図は本発明装置の構或を示すブロック図、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
ｆ８＋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ｆｅｌは本発明の加水分解ステージの気
化チャンバーの一実施例を示す断面図、第５図は本発明
の抽出・送液ステージの横或の一実施例を示す外観図、
第６図ｔａ＋〜（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 block diagram showing the structure of the apparatus of the present invention, 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 f8+ is an external view showing an embodiment of the hydrolysis stage of the present invention when the sample holder is released, and FIG. A sectional view showing an embodiment of the hydrolysis stage when the sample holder is fixed, FIG. 4 fel is a sectional view showing an embodiment of the vaporization chamber of the hydrolysis stage of the present invention, and FIG. An external view showing an embodiment of the liquid stage,
Figures 6 ta+ to d+ are cross-sectional views for explaining the operation of the transport mechanism of the sample holder of the present invention.
Claims (4)
セラミックス粒からなり、タンパク質あるいはペプチド
の試料を担持する試料保持部と、該試料保持部を支持す
るガラスもしくはセラミックスからなる外筒とで構成す
る試料担体と、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) Consisting of a sample holder made of porous glass, glass fiber, glass grains, or ceramic granules and supporting a protein or peptide sample, and an outer cylinder made of glass or ceramics that supports the sample holder. A sample carrier, a transportation mechanism for the sample carrier carrying a dried sample of protein or peptide, and a hydrolysis reaction through a solid phase-gas phase reaction by supplying acid mixed vapor to the dried sample under pressure and heat. a mechanism for transporting a sample carrier carrying a hydrolyzed sample; a mechanism for extracting the hydrolyzed sample from the sample carrier; and a sample solution injection mechanism for the extracted sample solution into an amino acid analyzer. 1. A device for hydrolyzing proteins or peptides, comprising: a mechanism for feeding liquid to the device; and a mechanism for supplying fluid used for the hydrolysis, extraction, and liquid feeding.
のそれよりも細くしてあ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を加水分解する装置。(2) The apparatus for hydrolyzing proteins or peptides according to claim 1, wherein the outer cylinder of the sample carrier has an outer diameter smaller at both ends than at the center.
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。(3) 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。(4) 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 |
|---|---|---|---|
| JP15080189A JPH0317048A (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 |
|---|---|---|---|
| JP15080189A JPH0317048A (en) | 1989-06-13 | 1989-06-13 | Device of hydrolyzing protein or peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0317048A true JPH0317048A (en) | 1991-01-25 |
Family
ID=15504727
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP15080189A Pending JPH0317048A (en) | 1989-03-17 | 1989-06-13 | Device of hydrolyzing protein or peptide |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0317048A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0317050U (en) * | 1989-07-03 | 1991-02-20 |
-
1989
- 1989-06-13 JP JP15080189A patent/JPH0317048A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0317050U (en) * | 1989-07-03 | 1991-02-20 |
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