JPS60107548A - Detecting method of bubble in column - Google Patents
Detecting method of bubble in columnInfo
- Publication number
- JPS60107548A JPS60107548A JP58214055A JP21405583A JPS60107548A JP S60107548 A JPS60107548 A JP S60107548A JP 58214055 A JP58214055 A JP 58214055A JP 21405583 A JP21405583 A JP 21405583A JP S60107548 A JPS60107548 A JP S60107548A
- Authority
- JP
- Japan
- Prior art keywords
- column
- light
- bubble
- gel
- photodetector
- 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
- 238000000034 method Methods 0.000 title claims description 9
- 238000004811 liquid chromatography Methods 0.000 claims description 5
- 230000007423 decrease Effects 0.000 abstract description 5
- 230000005540 biological transmission Effects 0.000 abstract 1
- 230000001678 irradiating effect Effects 0.000 abstract 1
- 239000000499 gel Substances 0.000 description 19
- 238000005259 measurement Methods 0.000 description 8
- 238000004587 chromatography analysis Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 239000011543 agarose gel Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000007863 gel particle Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/02—Investigating particle size or size distribution
- G01N15/0205—Investigating particle size or size distribution by optical means
Landscapes
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth 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)
- Geophysics And Detection Of Objects (AREA)
Abstract
Description
【発明の詳細な説明】
技術分野
本発明は、液体クロマトグラフィーに用いるゲルカラム
内に発生又は侵入する気泡を検出する方法に関するもの
である。DETAILED DESCRIPTION OF THE INVENTION Technical Field The present invention relates to a method for detecting air bubbles generated or entering a gel column used in liquid chromatography.
従来技術
ゲルカラムを用いる液体クロマトグラフィーで正確な測
定を行なうには、カラム内にゲル粒子を均一に充填する
と共にカラム内の気泡の発生や空気の侵入を防止するこ
とが極めて重要である。カラム内に気泡が発生すると、
溶液がカラム内を均一に流れずカラム内の反応が不均一
、不安定になったり、発生する気泡がカラム的にかけら
れる圧力に対して緩衝作用をおこしクロマトピークに再
現性がなくなる不具合が生ずる。従って、測定中に気泡
が発生すると測定値自体に信頼性がなくなるためゲルカ
ラム内の気泡の検出は極めて重要な課題になっている。In order to perform accurate measurements in liquid chromatography using conventional gel columns, it is extremely important to uniformly pack gel particles into the column and to prevent the formation of bubbles and the intrusion of air within the column. When air bubbles occur in the column,
Problems arise in that the solution does not flow uniformly within the column, making the reaction within the column uneven and unstable, and that the bubbles generated act as a buffer against the pressure applied to the column, resulting in a loss of reproducibility in chromatographic peaks. Therefore, if air bubbles are generated during measurement, the measured value itself becomes unreliable, so detection of air bubbles in the gel column has become an extremely important issue.
従来、ゲルカラム内の気泡の発生の検出方法として、ゲ
ルカラムに流す溶液の圧力変化を検出する方法が知られ
ている。このような方法は高圧条件の下ではゲルカラム
内の気泡の発生を検出できるが、低圧条件下で行なうク
ロマトグラフィーでは圧力変化が微小であるため気泡の
発生を正確に検出することは不可能である。BACKGROUND ART Conventionally, as a method for detecting the generation of air bubbles in a gel column, a method of detecting a pressure change in a solution flowing through a gel column is known. This method can detect the generation of air bubbles in a gel column under high pressure conditions, but in chromatography performed under low pressure conditions, it is impossible to accurately detect the generation of air bubbles because the pressure changes are minute. .
従って、ゲルカラムを用いてクロマトグラフィー測定を
行なう場合に気泡の発生を正確に検出できる方法の開発
が強く要請されている。Therefore, there is a strong demand for the development of a method that can accurately detect the generation of bubbles when performing chromatography measurements using gel columns.
発明の目的
本発明の目的は、液体クロマトグラフィーに用いるゲル
カラム内に発生又は侵入する気泡を正確に検出できるゲ
ルカラム内の気泡検出方法を提供することにある。OBJECTS OF THE INVENTION An object of the present invention is to provide a method for detecting air bubbles in a gel column that can accurately detect air bubbles generated or entering a gel column used for liquid chromatography.
本発明は、液体クロマトグラフィーに用いるゲルカラム
内に発生又は侵入する気泡を検出するに当り、ゲルカラ
ムに均一に光を投射し、カラムを透過した透過光を受光
素子で受光して、受光素子の出力変化によりゲルカラム
内の気泡を検出することを特徴とするものである。In detecting air bubbles generated or entering a gel column used for liquid chromatography, the present invention uniformly projects light onto the gel column, receives transmitted light that has passed through the column with a light receiving element, and outputs an output from the light receiving element. This method is characterized by detecting air bubbles in the gel column based on changes in the gel column.
第１図は本発明による気泡検出方法の一例の構成を示す
線図である。光源１から発した光を拡散板２により拡散
光とした後、フメリータレンズ３により平行光としてカ
ラム４に投射する。この入射光は拡散板２で拡散光にさ
れているからカラムにはほぼ全面に亘り均一の強度分布
の光束が入射する。カラム４は透明ガラス管５に分子数
８０〜１４０μｍのアガロースゲル６が均一に充填され
、更に溶液が上方より入り下方に流出するように構成さ
れている。カラム４に入射した光はカラム内のゲルによ
り均一に拡散され受光器７で受光される。受光器７は、
俵数の受光素子をカラム４の全長に亘って配列した構成
となっている。カラム４内に気泡が発生すると、気泡の
屈折率と溶液の屈折率が相異するため気泡表面で散乱反
射が生じ、気泡が存在する部分では光の透過量が著しく
減少して受光器７で受光される光量が減少する。気泡の
大きさはアガロースゲル５の大きざに比べはるかに大き
いため、気泡による透過光量の減少は受光器７により明
瞭に観測される。特に１受光器７を構成する受光素子の
受光面積を気泡の断面積とほぼ等しい大きさに設定すれ
ば、気泡の位置と対応する位置にある受光素子の光量変
化は非常に大きなものとなる。FIG. 1 is a diagram showing the configuration of an example of the bubble detection method according to the present invention. Light emitted from a light source 1 is converted into diffused light by a diffusion plate 2, and then projected onto a column 4 as parallel light by a fumerita lens 3. Since this incident light is diffused by the diffuser plate 2, a light beam with a uniform intensity distribution is incident on the column almost over the entire surface. The column 4 is constructed such that a transparent glass tube 5 is uniformly filled with agarose gel 6 having molecules of 80 to 140 μm, and the solution enters from above and flows out from below. The light incident on the column 4 is uniformly diffused by the gel within the column and is received by the light receiver 7. The light receiver 7 is
The structure has a number of bales of light receiving elements arranged over the entire length of the column 4. When air bubbles occur in the column 4, the refractive index of the air bubbles and the refractive index of the solution are different, so scattering reflection occurs on the bubble surface, and the amount of light transmitted in the area where the air bubbles exist is significantly reduced, causing light to be transmitted to the light receiver 7. The amount of light received decreases. Since the size of the bubbles is much larger than the size of the agarose gel 5, the decrease in the amount of transmitted light due to the bubbles is clearly observed by the light receiver 7. In particular, if the light-receiving area of the light-receiving element constituting one light receiver 7 is set to be approximately equal to the cross-sectional area of the bubble, the change in the light amount of the light-receiving element located at the position corresponding to the bubble will be very large.
第２図は受光器７の出力の信号処理回路の一例の構成を
示すブロック図である。受光器７を構成する各受光素子
の出力端子８からの出力信号ａ〜ｍを加算器９に入力さ
せ、各受光素子の出力の和を比較器１０の一方の端子に
入力する。比較器１０の他方の端子には基準電圧ｖｒを
印加し受光器７の出力電圧と基準電圧ｖｒとを比較する
。この基準電圧は１各種の測定溶液及び各種の充填する
ゲル毎に気泡のない状態での受光器７の出力電圧をあら
かじめ測定しておき定めることができる。このように構
成すれば、クロマトグラフィー測定中にカラム４内のい
ずれかの箇所で気泡が発生しても、受光器７の出力信号
が比較器７で基準電圧と比較され、比較器７の出力変動
により即時に気泡の発生が検知される。そして、比較器
７の出力信号により気泡の混入を表示しクロマトグラフ
ィーの測定を一時停止することにより異常値の測定を未
然に防止することができる。FIG. 2 is a block diagram showing the configuration of an example of a signal processing circuit for outputting the light receiver 7. As shown in FIG. Output signals a to m from the output terminals 8 of the light receiving elements constituting the light receiver 7 are input to an adder 9, and the sum of the outputs of the respective light receiving elements is input to one terminal of the comparator 10. A reference voltage vr is applied to the other terminal of the comparator 10, and the output voltage of the light receiver 7 and the reference voltage vr are compared. This reference voltage can be determined by measuring in advance the output voltage of the light receiver 7 in a bubble-free state for each type of measurement solution and each type of gel to be filled. With this configuration, even if bubbles occur anywhere in the column 4 during chromatography measurement, the output signal of the photodetector 7 is compared with the reference voltage in the comparator 7, and the output of the comparator 7 is The occurrence of bubbles is immediately detected due to fluctuations. The output signal of the comparator 7 indicates the presence of bubbles and the chromatography measurement is temporarily stopped, thereby making it possible to prevent abnormal values from being measured.
尚、本例ではカラム５の全長に亘って複数の受光素子を
配列したがカラム５の全長を補える太きさの１個の受光
素子で検出する構成としてもよい０また、カラム４の前
面にカラム５の全長に対応した長さを有し、カラム管径
より小さい幅を有するスリット開口をあけたスリット板
を配置すれば、カラムの周辺を通過する妨害光をカット
でき、受光器７における検出精度を高めることができる
０更に、本例では信号処理回路の構成として、複数の受
光素子からの出力信号の和を基準電圧と比較する構成と
したが、この信号処理回路の構成は幾多の変形が可能で
ある。例えば、各受光素子の出力信号毎に基準電圧と比
較する構成によれば〜透過光量の減少をより正確に検出
することが可能になる。又、上述した実施例では充填ゲ
ルとしてアガロースゲルを用いたがアガロースゲルに限
定されるものではなく、クロマトグラフィー用の各種の
ゲルにも適用できる。In this example, a plurality of light receiving elements are arranged over the entire length of the column 5, but a configuration in which detection is performed with a single light receiving element of a thickness that can compensate for the entire length of the column 5 may also be used. By arranging a slit plate with a slit opening having a length corresponding to the entire length of the column 5 and a width smaller than the column tube diameter, interference light passing around the column can be cut, and detection by the light receiver 7 can be prevented. Furthermore, in this example, the configuration of the signal processing circuit is such that the sum of output signals from a plurality of light receiving elements is compared with a reference voltage, but the configuration of this signal processing circuit can be modified in many ways. is possible. For example, if the configuration is such that each output signal of each light receiving element is compared with a reference voltage, it becomes possible to more accurately detect a decrease in the amount of transmitted light. Furthermore, although agarose gel was used as the filling gel in the above embodiments, the present invention is not limited to agarose gel, and can also be applied to various gels for chromatography.
発明の詳細
な説明したように本発明によれば、簡単な構成でゲルカ
ラム内の気泡の発生を正確に検知することができ、低圧
条件下で行なうクロマトグラフィーの測定中における異
常な測定値の釦生を未然に防止し正確な測定データの採
取が可能になる０DETAILED DESCRIPTION OF THE INVENTION According to the present invention, it is possible to accurately detect the occurrence of air bubbles in a gel column with a simple configuration, and it is possible to detect abnormal measured values during chromatography measurements performed under low pressure conditions. This makes it possible to prevent errors and collect accurate measurement data.
第１図は本発明による気泡検出方法の一例の構成を示す
線図、
第２図は本発明の信号処理回路の一例の構成を示すプシ
ツク図である。
１・・・光源 ２・・・拡散板
８・・・コリメータレンズ ４・・・ゲルカラム５…ガ
ラス管 ６・・・アガ田−スゲル７・・・受光器 ８・
・・出力端子
９・・・加算器 １ｏ・・・比較器FIG. 1 is a diagram showing the configuration of an example of the bubble detection method according to the present invention, and FIG. 2 is a diagram showing the configuration of an example of the signal processing circuit of the present invention. 1... Light source 2... Diffusion plate 8... Collimator lens 4... Gel column 5... Glass tube 6... Agada-Sgel 7... Light receiver 8.
...Output terminal 9...Adder 1o...Comparator
Claims (1)
又は侵入する気泡を検出するに当たり、ゲルカラムに均
一に光を投射し、カラムを透過した透過光を受光素子で
受光して、受光素子の出力変化によりゲルカラム内の気
泡を検出することを特徴とするカラム内の気泡を検出す
る方法。L To detect air bubbles generated or entering a gel column used for liquid chromatography, light is uniformly projected onto the gel column, the transmitted light transmitted through the column is received by a light receiving element, and the change in the output of the light receiving element is used to detect bubbles inside the gel column. A method for detecting air bubbles in a column, comprising detecting air bubbles.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58214055A JPS60107548A (en) | 1983-11-16 | 1983-11-16 | Detecting method of bubble in column |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58214055A JPS60107548A (en) | 1983-11-16 | 1983-11-16 | Detecting method of bubble in column |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS60107548A true JPS60107548A (en) | 1985-06-13 |
Family
ID=16649511
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP58214055A Pending JPS60107548A (en) | 1983-11-16 | 1983-11-16 | Detecting method of bubble in column |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS60107548A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016217857A (en) * | 2015-05-20 | 2016-12-22 | 株式会社ネクスコ東日本エンジニアリング | Detection device and detection method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4915402U (en) * | 1972-05-15 | 1974-02-08 | ||
| JPS543759A (en) * | 1977-06-10 | 1979-01-12 | Hitachi Ltd | Machine hand |
-
1983
- 1983-11-16 JP JP58214055A patent/JPS60107548A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4915402U (en) * | 1972-05-15 | 1974-02-08 | ||
| JPS543759A (en) * | 1977-06-10 | 1979-01-12 | Hitachi Ltd | Machine hand |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2016217857A (en) * | 2015-05-20 | 2016-12-22 | 株式会社ネクスコ東日本エンジニアリング | Detection device and detection method |
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