JPH08201372A - Coloration method by ninhydrin reaction - Google Patents
Coloration method by ninhydrin reactionInfo
- Publication number
- JPH08201372A JPH08201372A JP1397895A JP1397895A JPH08201372A JP H08201372 A JPH08201372 A JP H08201372A JP 1397895 A JP1397895 A JP 1397895A JP 1397895 A JP1397895 A JP 1397895A JP H08201372 A JPH08201372 A JP H08201372A
- Authority
- JP
- Japan
- Prior art keywords
- ninhydrin
- solution
- compound
- inspected
- detected
- 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
Landscapes
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明はニンヒドリン反応による
呈色方法に関し、さらに詳しくは迅速処理に適し高感度
なニンヒドリン反応による呈色方法に関するものであ
る。特に、カスガノビオサミン誘導体の検出方法に関す
る。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a ninhydrin reaction coloration method, and more particularly to a highly sensitive ninhydrin reaction coloration method suitable for rapid processing. In particular, it relates to a method for detecting a kasuganobiosamine derivative.
【０００２】[0002]
【従来の技術】ニンヒドリン反応は蛋白質の呈色反応の
一つであり、アミノ酸、ポリペプチド、蛋白質だけでな
く、アミノ基を有する有機化合物やイミノ化合物あるい
は核酸塩基の検出用の試薬としても使われている。溶液
中の被検物質の定量、定性のほかに、濾紙クロマトグラ
フや薄層クロマトグラフ上の定性、定量にも簡便に用い
られる。BACKGROUND OF THE INVENTION The ninhydrin reaction is one of the color reactions of proteins and is used not only as amino acids, polypeptides and proteins, but also as reagents for detecting organic compounds having amino groups, imino compounds or nucleobases. ing. In addition to quantitative and qualitative analysis of the test substance in the solution, it can be easily used for qualitative and quantitative analysis on filter paper chromatographs and thin-layer chromatographs.
【０００３】ニンヒドリンと披験化合物との呈色反応は
遅く、迅速に検出するには従来は加熱、加温することが
必要であった。すなわち、溶液中の化合物の検出には、
ニンヒドリン溶液を添加した後煮沸し呈色反応を充分行
わせ次いで溶液を冷却する事が必要であった。また、濾
紙クロマトグラフや薄層クロマトグラフ上の化合物の検
出においても、ニンヒドリンとの接触後の60〜100℃で
の加熱が迅速な呈色反応には必要であった。しかしなが
ら、加熱操作そのものの煩雑性や、不均一な加熱による
むらや定量の欠如がこれらの従来法では問題となってい
た。The color reaction between ninhydrin and the test compound is slow, and heating and heating were conventionally required for rapid detection. That is, for the detection of compounds in solution,
After adding the ninhydrin solution, it was necessary to boil it to allow the color reaction to sufficiently occur, and then cool the solution. Also, in the detection of compounds on filter paper chromatographs and thin-layer chromatographs, heating at 60-100 ° C after contact with ninhydrin was necessary for rapid color reaction. However, the complexity of the heating operation itself, unevenness due to uneven heating, and lack of quantification have been problems in these conventional methods.
【０００４】[0004]
【発明が解決しようとする課題】本発明は、このような
問題点を解決し、加熱操作なしで迅速、簡便に、しかも
高感度にニンヒドリンと被験化合物、特にカスガノビオ
サミン誘導体との呈色反応を行わせることを目的とする
方法である。DISCLOSURE OF THE INVENTION The present invention solves the above problems and provides a color reaction between ninhydrin and a test compound, particularly a kasuganobiosamine derivative, rapidly, simply and with high sensitivity without a heating operation. It is a method for the purpose of letting you do.
【０００５】[0005]
【課題を解決するための手段】本発明の上記目的は、ニ
ンヒドリン反応を、紫外線の照射下で行うことを特徴と
するニンヒドリン反応による呈色方法により達成され
る。The above-mentioned object of the present invention is achieved by a ninhydrin reaction-based coloring method characterized in that the ninhydrin reaction is carried out under irradiation of ultraviolet rays.
【０００６】好ましくは、前記紫外線の波長が400nm以
下であり、更に好ましくは前記紫外線の波長が200〜380
nmである。The wavelength of the ultraviolet rays is preferably 400 nm or less, more preferably the wavelength of the ultraviolet rays is 200 to 380.
nm.
【０００７】本発明の呈色反応について更に詳しく説明
する。The color reaction of the present invention will be described in more detail.
【０００８】本発明の溶液中のニンヒドリン呈色反応
は、被験化合物に対し過剰量のニンヒドリンを添加し行
われる。一般的には、好ましくは0.1〜５％のニンヒド
リン溶液を被検化合物の溶解された溶液に対し１／100
〜１／10容加える。紫外光を一定時間照射し生成した生
成色素を検出する。アミノ酸、ポリペプチド、アミン化
合物、カスガノビオサミン誘導体の場合は最大吸収波長
が570nm前後、イミン化合物の場合は440nm前後、核酸塩
基の場合は250nm前後である。The ninhydrin color reaction in the solution of the present invention is carried out by adding an excessive amount of ninhydrin to the test compound. Generally, 0.1 to 5% ninhydrin solution is preferably added to 1/100 of the solution of the test compound.
Add ~ 1/10 volume. Ultraviolet light is radiated for a certain period of time to detect the formed dye. The maximum absorption wavelength is around 570 nm for amino acids, polypeptides, amine compounds, and kasuganobiosamine derivatives, around 440 nm for imine compounds, and around 250 nm for nucleobases.
【０００９】本発明に特に適用できる被倹物質は、カス
ガノビオサミン誘導体である。カスガノビオサミン誘導
体の例としては、カスガノビオサミン、カスガマイシ
ン、N2-CBZ-カスガノビオサミン及びこれらの塩が挙げ
られる。カスガマイシンはアミノグリコシド抗生物質で
あり、稲のいもち病の防除剤として用いられる。カスガ
ノビオサミンは、Advan.Chem.Ser.;74,15-40(1968)記載
の方法により、カスガマイシンのアルカリ加水分解によ
り合成できる。カスガノビオサミン誘導体については、
特開昭46-28593号、同46-28594号、Bull.Inst.Chem.Re
s.,Kyoto Univ.;50(3),275-302(1972)などに記載されて
いる。The target substance particularly applicable to the present invention is a kasuganobiosamine derivative. Examples of the kasuganobiosamine derivative include kasuganobiosamine, kasugamycin, N 2 -CBZ-cassanobiosamine, and salts thereof. Kasugamycin is an aminoglycoside antibiotic and is used as a control agent for rice blast disease. Kasuganobiosamine can be synthesized by alkaline hydrolysis of kasugamycin by the method described in Advan. Chem. Ser .; 74 , 15-40 (1968). For kasuganobiosamine derivatives,
JP-A-46-28593, JP-A-46-28594, Bull.Inst.Chem.Re
s., Kyoto Univ .; 50 (3), 275-302 (1972) and the like.
【００１０】本発明において、濾紙クロマトグラフや薄
層クロマトグラフ上の化合物の検出には、まずニンヒド
リン溶液と接触させる。接触は濾紙や薄層をニンヒドリ
ン溶液に浸漬させるか、噴霧で行われるが、噴霧するこ
とが好ましい。0.1〜５％のニンヒドリン溶液（溶解液
としてはアセトンやアルコール類が好ましい）を噴霧し
た後風乾し、次いで紫外光を一定時間照射した後、生成
した色素を検出する。In the present invention, in detecting a compound on a filter paper chromatograph or a thin layer chromatograph, first, a compound is brought into contact with a ninhydrin solution. The contact is performed by immersing the filter paper or thin layer in a ninhydrin solution or by spraying, but spraying is preferable. A 0.1 to 5% ninhydrin solution (acetone or alcohol is preferable as a solution) is sprayed, air-dried, and then irradiated with ultraviolet light for a certain period of time, and then the produced dye is detected.
【００１１】本発明で使用できる紫外光は波長400nm以
下の光である。好ましくは200nmから380nmの光である。
紫外光の照射時間と光強度は呈色反応が充分行われる範
囲であればよく、それぞれの兼ね合いで変化する。濾紙
クロマトグラフや薄層クロマトグラフ上の化合物の検出
には、光強度0.5〜１mW／cm3程度の一般のＵＶランプで
あれば２〜10分間で充分である。The ultraviolet light that can be used in the present invention is light having a wavelength of 400 nm or less. Light of 200 nm to 380 nm is preferable.
The irradiation time and the light intensity of the ultraviolet light may be in the range where the color reaction is sufficiently carried out, and change depending on the respective balance. For detection of compounds on filter paper chromatographs and thin-layer chromatographs, a general UV lamp with a light intensity of 0.5 to 1 mW / cm 3 is sufficient for 2 to 10 minutes.
【００１２】[0012]
【実施例】以下、実施例を挙げて本発明を詳細に説明す
るが、本発明の態様はこれに限定されない。The present invention will be described in detail below with reference to examples, but the embodiments of the present invention are not limited thereto.
【００１３】実施例１ Whatman3MM濾紙上に200ug／mlの濃度のカスガマイシン
塩酸塩水溶液を５ul点着した。次いで、この濾紙にニン
ヒドリンの0.5％エタノール溶液を均一に噴霧した。風
乾後、紫外線ランプを用い365nmの紫外線（光強度0.61m
W／cm3）を照射した。10分後には青紫色のスポットが確
認できた。一方、紫外線を照射しなかった場合は、１時
間後でもスポットは確認できなかった。Example 1 On a Whatman 3MM filter paper, 5ul of an aqueous solution of kasugamycin hydrochloride having a concentration of 200ug / ml was spotted. Then, a 0.5% ethanol solution of ninhydrin was uniformly sprayed on the filter paper. After air-drying, use an ultraviolet lamp to emit 365 nm ultraviolet light (light intensity 0.61 m
W / cm 3 ) was irradiated. After 10 minutes, a blue-violet spot was confirmed. On the other hand, when ultraviolet rays were not irradiated, no spots could be confirmed even after 1 hour.
【００１４】実施例２ N2-CBZ-カスガノビオサミンを100ug／mlとなるよう純水
に溶解し、この２ulをシリカゲルTLCプラスチックシー
ト（メルク社製）に点着した。次いで、このシートにニ
ンヒドリンの0.3％アセトン溶液を均一に噴霧した。風
乾後、紫外線ランプを用いて312nmの紫外線（光強度0.5
8mW／cm3）を照射した。５分後には青紫色のスポットが
確認できた。一方、紫外線を照射しなかった場合は、１
時間後でもスポットは確認できなかった。Example 2 N 2 -CBZ-cassuganobiosamine was dissolved in pure water to 100 ug / ml, and 2 ul of this was spotted on a silica gel TLC plastic sheet (Merck). Then, the sheet was uniformly sprayed with a 0.3% acetone solution of ninhydrin. After air-drying, use an ultraviolet lamp to emit 312 nm ultraviolet light (light intensity 0.5
It was irradiated with 8 mW / cm 3 ). After 5 minutes, a blue-violet spot could be confirmed. On the other hand, if no ultraviolet ray was irradiated, 1
The spot could not be confirmed even after hours.
【００１５】[0015]
【発明の効果】本発明により、加熱操作なしで迅速、簡
便に、しかも高感度にニンヒドリンと被験化合物との呈
色反応を行わせることができる。EFFECTS OF THE INVENTION According to the present invention, a color reaction between ninhydrin and a test compound can be carried out rapidly, simply and highly sensitively without heating.
Claims (4)
行うことを特徴とするニンヒドリン反応による呈色方
法。1. A method of coloring by a ninhydrin reaction, which comprises performing the ninhydrin reaction under irradiation of ultraviolet rays.
とを特徴とする請求項１記載のニンヒドリン反応による
呈色方法。2. The coloration method by the ninhydrin reaction according to claim 1, wherein the wavelength of the ultraviolet rays is 400 nm or less.
とを特徴とする請求項１記載のニンヒドリン反応による
呈色方法。3. The coloration method by the ninhydrin reaction according to claim 1, wherein the wavelength of the ultraviolet rays is 200 to 380 nm.
項１、２又は３記載の呈色方法で行うことを特徴とする
ニンヒドリン反応による呈色方法。4. A coloring method by a ninhydrin reaction, which comprises detecting the kasuganobiosamine derivative by the coloring method according to claim 1, 2 or 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1397895A JPH08201372A (en) | 1995-01-31 | 1995-01-31 | Coloration method by ninhydrin reaction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1397895A JPH08201372A (en) | 1995-01-31 | 1995-01-31 | Coloration method by ninhydrin reaction |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH08201372A true JPH08201372A (en) | 1996-08-09 |
Family
ID=11848322
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1397895A Pending JPH08201372A (en) | 1995-01-31 | 1995-01-31 | Coloration method by ninhydrin reaction |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH08201372A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004507741A (en) * | 2000-08-24 | 2004-03-11 | バイオ−ラッド ラボラトリーズ，インコーポレイティド | Dry powder formulation for labeling proteins |
| JP2017517744A (en) * | 2014-06-13 | 2017-06-29 | エーエーエー サイエンティフィック リミテッド | Apparatus and method for preparing ninhydrin reagent |
-
1995
- 1995-01-31 JP JP1397895A patent/JPH08201372A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004507741A (en) * | 2000-08-24 | 2004-03-11 | バイオ−ラッド ラボラトリーズ，インコーポレイティド | Dry powder formulation for labeling proteins |
| JP2017517744A (en) * | 2014-06-13 | 2017-06-29 | エーエーエー サイエンティフィック リミテッド | Apparatus and method for preparing ninhydrin reagent |
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