JPS6342221B2 - - Google Patents
Info
- Publication number
- JPS6342221B2 JPS6342221B2 JP14570084A JP14570084A JPS6342221B2 JP S6342221 B2 JPS6342221 B2 JP S6342221B2 JP 14570084 A JP14570084 A JP 14570084A JP 14570084 A JP14570084 A JP 14570084A JP S6342221 B2 JPS6342221 B2 JP S6342221B2
- Authority
- JP
- Japan
- Prior art keywords
- information
- scanning
- memory
- scanning stage
- thin layer
- 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.)
- Expired
Links
- 230000003287 optical effect Effects 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 9
- 238000005375 photometry Methods 0.000 claims description 5
- 230000001360 synchronised effect Effects 0.000 claims description 2
- 238000004809 thin layer chromatography Methods 0.000 claims 1
- 239000000523 sample Substances 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 3
- 238000005070 sampling Methods 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000010365 information processing Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
Landscapes
- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Description
【発明の詳細な説明】
本発明は薄層クロマトグラフにおける薄層プレ
ートの分光吸光度測定に二波長方式を用いる場合
の光走査方法に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an optical scanning method when a two-wavelength method is used to measure the spectral absorbance of a thin layer plate in a thin layer chromatograph.
薄層クロマトグラフの薄層プレートの吸光度測
定において薄層プレートの厚さむら等によるノイ
ズを除去するため二波長方式が用いられるが、従
来はこの二波長方式を実行するために互に異なる
波長の光を出すように設定された２つの分光器を
用い、その両分光器の射出光をセクターミラーを
用いて交互に試料に入射させるようにしていたか
ら、分光器が２台必要であり、またセクターミラ
ー及びその駆動機構が必要で更に２つの分光器か
らの光を同一光路に導くための光学系が必要であ
るため装置は高価複雑なものとなつていた。 When measuring the absorbance of thin-layer plates in thin-layer chromatographs, a dual-wavelength method is used to remove noise caused by uneven thickness of the thin-layer plate. Two spectrometers were used that were set to emit light, and the emitted light from both spectrometers was made to enter the sample alternately using a sector mirror, so two spectrometers were required, and a sector mirror was also used. The device is expensive and complicated because it requires a drive mechanism and an optical system for guiding the light from the two spectrometers to the same optical path.
本発明は薄層プレートの光走査装置で分光器が
１つでよくセクターミラー及びそれに付随する部
分及び２個所から来る光線を同一光路に導入する
光学系が不要な光学走査方法を提供することを目
的としている。 The present invention provides an optical scanning method using a thin-layer plate optical scanning device, which requires only one spectroscope and does not require a sector mirror, an associated portion thereof, and an optical system that introduces light beams coming from two places into the same optical path. The purpose is
本発明は一定波長の光で薄層プレートを走査
し、そのときの測光出力をメモリに記憶させてお
き、次に他の一定波長で薄層プレートを走査し、
先の記憶結果を読出して２回目の走査における測
光値との間で演算を行うことにより二波長方式を
用いたのと同じ結果を得ようとするものである。
以下実施例によつて本発明を説明する。 The present invention scans a thin layer plate with light of a fixed wavelength, stores the photometric output at that time in a memory, and then scans the thin layer plate with another fixed wavelength,
By reading out the previously stored result and performing calculations between it and the photometric value from the second scan, the same result as using the two-wavelength method is obtained.
The present invention will be explained below with reference to Examples.
第１図は本発明の一実施例を示す。１は光源、
２は分光器、３は走査ステージ、４は走査ステー
ジを駆動するパルスモータである。走査ステージ
３上に試料の薄層プレートがセツトされる。光源
１から出た光は分光器２に入射し、分光器２によ
つて単一波長λ１の光のみが分光器２から射出し
て走査ステージ３上の試料に入射せしめられる。
走査ステージ３を駆動するパルスモータは図では
４と番号をつけた一つだけが示されているが、具
体的には走査ステージをＸ方向Ｙ方向に駆動し得
るよう二個のパルスモータがある。５は光検出器
で試料の薄層プレートの透過光或は反射光を受光
し、電気信号に変換する。この電気信号は増幅器
６で増幅された後Ａ−Ｄ変換器１３でデイジタル
信号に変換される。７はマイクロプロセツサであ
る。パルスモータ４は制御回路８から供給される
パルスで駆動せしめられ、制御回路８はマイクロ
プロセツサ７からの指令により正転或は逆転パル
スをパルスモータ４に印加すると共に同じパルス
をマイクロプロセツサに送る。マイクロプロセツ
サはこのパルスを計数して走査ステーズ３の位置
の信号を得る。Ａ−Ｄ変換器１３はマイクロプロ
セツサ７よりパルスモータ４を駆動するパルスと
同期したサンプリング信号を受けて増幅器６の出
力をサンプリングしこれをＡ−Ｄ変換し、このＡ
−Ｄ変換出力はマイクロプロセツサ７を介してラ
ンダムアクセスメモリ（RAM）９に書込まれ
る。このときRAMのアドレスを指定する情報は
マイクロプロセツサ７が制御回路８から受取つた
パルスを計数した値であつて、RAMのアドレス
は薄層プレート上の位置に対応している。１０は
マイクロプロセツサ７の動作プログラムが書込ん
である読出し専用メモリ（ROM）である。 FIG. 1 shows an embodiment of the invention. 1 is a light source,
2 is a spectrometer, 3 is a scanning stage, and 4 is a pulse motor that drives the scanning stage. A thin layer plate of a sample is set on the scanning stage 3. Light emitted from a light source 1 is incident on a spectroscope 2, and only light with a single wavelength λ1 is emitted from the spectrometer 2 and is made to enter a sample on a scanning stage 3.
Only one pulse motor numbered 4 is shown in the figure for driving the scanning stage 3, but specifically there are two pulse motors that can drive the scanning stage in the X and Y directions. . A photodetector 5 receives transmitted light or reflected light from the thin plate of the sample and converts it into an electrical signal. This electrical signal is amplified by an amplifier 6 and then converted into a digital signal by an A/D converter 13. 7 is a microprocessor. The pulse motor 4 is driven by pulses supplied from the control circuit 8, and the control circuit 8 applies forward or reverse rotation pulses to the pulse motor 4 according to commands from the microprocessor 7, and also applies the same pulses to the microprocessor. send. The microprocessor counts these pulses to obtain a signal for the position of the scanning stage 3. The A-D converter 13 receives a sampling signal synchronized with the pulse that drives the pulse motor 4 from the microprocessor 7, samples the output of the amplifier 6, converts it A-D,
-D conversion output is written to random access memory (RAM) 9 via microprocessor 7. At this time, the information specifying the RAM address is a value obtained by counting the pulses received by the microprocessor 7 from the control circuit 8, and the RAM address corresponds to a position on the thin layer plate. Reference numeral 10 denotes a read-only memory (ROM) in which an operating program for the microprocessor 7 is written.
マイクロプロセツサ７は次のような動作をす
る。装置をスタートさせると試料ステージ３を走
査の始点に移動させると共にパルスモータ１１を
操作して回折格子Ｇを予め定めてある波長λ１の
位置にセツトし、走査ステージ３に走査運動を行
わせる。波長λ１は薄層プレートに対しても同プ
レート上に展開されている試料物質に対しても同
じような吸収を与える波長に選んであり、測光結
果は薄層プレート自体のむらの情報であつてこれ
がRAM１０にメモリされる。一回の走査が終る
と走査ステージを走査始点に戻しパルスモータ１
１を制御して分光器２の波長をλ２にセツトし再
び走査を開始させる。波長λ２は薄層プレート上
に展開されている物質によつて特に吸収される波
長に選んである。測光出力はＡ−Ｄ変換器でサン
プリングされデイジタル信号に変換されるがサン
プリングの度にRAMからそのときのステージ３
の位置に対応するアドレスの記憶を読出し上記デ
イジタル信号と共に演算を行い測光結果から薄層
プレートのむら等に基くノイズ成分を除く。この
ノイズ成分を除去された正しい情報は表示装置１
２に出力されてプリントされ、或はＤ−Ａ変換さ
れてカーブとして描記される。 The microprocessor 7 operates as follows. When the apparatus is started, the sample stage 3 is moved to the scanning starting point, and the pulse motor 11 is operated to set the diffraction grating G at a predetermined wavelength λ1 position, and the scanning stage 3 is caused to perform a scanning movement. The wavelength λ1 was selected to give similar absorption to both the thin layer plate and the sample material developed on the same plate, and the photometry result is information on the unevenness of the thin layer plate itself. It is stored in RAM10. When one scan is completed, the scanning stage is returned to the scanning starting point and the pulse motor 1
1 to set the wavelength of the spectrometer 2 to λ2 and start scanning again. The wavelength λ2 is chosen to be one that is specifically absorbed by the material spread out on the thin layer plate. The photometric output is sampled by an A-D converter and converted to a digital signal, but each time it is sampled, the current stage 3
The memory of the address corresponding to the position is read out and arithmetic is performed together with the digital signal to remove noise components based on unevenness of the thin plate and the like from the photometry results. The correct information from which this noise component has been removed is displayed on the display device 1.
2 and printed, or DA converted and drawn as a curve.
薄層プレートは第２図に示すように複数チヤン
ネルに展開されており、走査ステージ３はＸ、Ｙ
両方向に駆動されるようになつていてマイクロプ
ロセツサ７からの指令でパルスモータを介して走
査レーンの切換えがなされ、走査軌跡は図にＳで
示すような形となり、イの部分は速送りであり、
ロの部分で測光が行われる。 The thin layer plate is developed into multiple channels as shown in Fig. 2, and the scanning stage 3 is
It is designed to be driven in both directions, and the scanning lane is switched via a pulse motor in response to a command from the microprocessor 7, and the scanning trajectory is shaped as shown in the figure by S, with the part A being a fast feed. can be,
Photometry is performed in the area shown in (b).
マイクロプロセツサ４は与えられるプログラム
によりRAM１０の記憶情報及び現に測光しつゝ
ある情報等を用い、単にノイズ除去のみでなく他
の種々な情報処理が可能である。例えば微分動作
で現在得られた情報とそれより少し以前例えば前
回サンプリング時の情報（RAMに入れてある）
との差を求めれば微分情報が得られ吸光度の記録
だけでは明瞭に分離できないような二つのピーク
が分離可能となり、また同一波長を用いて試料の
展開されているレーンとその中間のレーンとを走
査することにより両者の差で薄層プレートの汚れ
とか曲り等のゆつくりした変化による測光結果の
ベースラインの変動を消去するような動作をさせ
ることもできる。 The microprocessor 4 uses information stored in the RAM 10, information currently being measured, etc. according to a given program, and is capable of performing not only noise removal but also various other information processing. For example, the information currently obtained by the differential operation and the information a little earlier, for example, from the previous sampling (stored in RAM)
By calculating the difference between the two peaks, differential information can be obtained, making it possible to separate two peaks that cannot be clearly separated by recording absorbance alone.Also, using the same wavelength, it is possible to separate the lane in which the sample is developed and the lane in between. By scanning, the difference between the two can be used to eliminate fluctuations in the baseline of photometry results due to slow changes such as dirt or bending of the thin layer plate.
本発明光走査方法は上述したような構成で２波
長方式を用いる場合分光器が１台でよく、光学系
が簡単になり、従つて装置構成が安価である。ま
たメモリのアドレスの指定に走査ステージ上の光
照射位置のデータを検出してこれをアドレス指定
情報としているから、二回の走査における測定値
の位置の対応が正確に得られるので、二波長波の
特長である薄層自体に基くノイズ除去が完全に行
われる。なおマイクロプロセツサを利用すると自
動化と共にプログラムの組み方により２波長方式
以外の種々な動作及びデータ処理ができる。 When the optical scanning method of the present invention uses the two-wavelength method with the above-described configuration, only one spectroscope is required, the optical system is simple, and the device configuration is therefore inexpensive. In addition, since the memory address is specified by detecting the data of the light irradiation position on the scanning stage and using this as the addressing information, it is possible to obtain accurate correspondence between the positions of the measured values in the two scans. Noise removal based on the thin layer itself is completely performed. Note that when a microprocessor is used, it is possible to perform various operations and data processing other than the two-wavelength method depending on automation and how the program is assembled.
第１図は本発明の一実施例装置の構成を示すブ
ロツク図、第２図は薄層プレート上の走査線を示
す平面図である。
１……光源、２……分光器、３……走査ステー
ジ、４……パルスモータ、１２……表示装置。
FIG. 1 is a block diagram showing the configuration of an apparatus according to an embodiment of the present invention, and FIG. 2 is a plan view showing scanning lines on a thin layer plate. DESCRIPTION OF SYMBOLS 1...Light source, 2...Spectroscope, 3...Scanning stage, 4...Pulse motor, 12...Display device.
Claims (1)
個の分光器と走査ステージ上の光照射位置の駆動
と同期した信号により走査ステージの位置の情報
を作り、これによつてメモリのアドレスを指定し
て一回目の走査における測光結果を記憶せしめ、
二回目の走査において、上記同様に位置の情報を
作り、これによつて上記メモリのアドレスを指定
して上記記憶を読出しこの記憶と現在得られた測
光データを用いて演算を行うことを特徴とする薄
層クロマトグラフイ用光走査方法。1. Information on the position of the scanning stage is created using a single spectrometer that projects light for sample irradiation onto the scanning stage and a signal that is synchronized with the drive of the light irradiation position on the scanning stage, and the address of the memory is specified using this information. to store the photometry results from the first scan,
In the second scan, position information is created in the same manner as above, and the address of the memory is specified using this information, the memory is read out, and calculations are performed using this memory and the currently obtained photometric data. An optical scanning method for thin layer chromatography.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14570084A JPS6046445A (en) | 1984-07-12 | 1984-07-12 | Optical scanning method for thin layer chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14570084A JPS6046445A (en) | 1984-07-12 | 1984-07-12 | Optical scanning method for thin layer chromatography |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6046445A JPS6046445A (en) | 1985-03-13 |
| JPS6342221B2 true JPS6342221B2 (en) | 1988-08-22 |
Family
ID=15391083
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14570084A Granted JPS6046445A (en) | 1984-07-12 | 1984-07-12 | Optical scanning method for thin layer chromatography |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6046445A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH087138B2 (en) * | 1987-02-14 | 1996-01-29 | 株式会社島津製作所 | Illumination mechanism for chromatographs |
-
1984
- 1984-07-12 JP JP14570084A patent/JPS6046445A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6046445A (en) | 1985-03-13 |
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