WO2016194108A1 - 分取クロマトグラフ - Google Patents
分取クロマトグラフ Download PDFInfo
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- WO2016194108A1 WO2016194108A1 PCT/JP2015/065796 JP2015065796W WO2016194108A1 WO 2016194108 A1 WO2016194108 A1 WO 2016194108A1 JP 2015065796 W JP2015065796 W JP 2015065796W WO 2016194108 A1 WO2016194108 A1 WO 2016194108A1
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- flow path
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/80—Fraction collectors
- G01N30/82—Automatic means therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/84—Preparation of the fraction to be distributed
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N2030/382—Flow patterns flow switching in a single column
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
Definitions
- the present invention relates to a preparative chromatograph in which a target component separated by a liquid chromatograph column is collected by a fraction collector.
- the preparative chromatograph is composed of a liquid chromatograph section, a detector and a fraction collector provided in the subsequent stage, and a control section for controlling these operations.
- a preparative chromatograph components in a sample separated and eluted in time by a column in a liquid chromatograph part are detected when passing through a detector, introduced into a fraction collector and collected in a preparative container (for example, Patent Documents 1 and 2).
- the liquid chromatograph section is composed of, for example, a liquid feed pump, a sample injection section, and a column, and components eluted from the column are introduced into a detector flow cell such as an absorption spectrophotometer through a pipe.
- a detector flow cell such as an absorption spectrophotometer
- a light source such as a deuterium lamp, a diffraction grating, and a motor for driving the diffraction grating are housed in a single casing, and the components that have passed through the flow cell are introduced into a fraction collector through a pipe.
- the separation flow path to which the sorting container such as a vial is connected, the waste liquid flow path to which the waste liquid container is connected, and the component that has passed through the detector are selected as the separation flow path or the waste liquid flow path.
- a flow path switching unit and the like that are circulated are accommodated in one housing.
- the time required for the target component to reach the flow channel switching unit of the fraction collector from the flow cell (delay time) is taken into consideration.
- the flow path switching unit is switched, and the target component is collected in the sorting container. Specifically, when the delay time has elapsed from the start time of detection of the target component, the flow path switching unit switches the flow path to the sorting flow path side, and sampling of the target component is started. When the delay time elapses, the flow path switching unit switches the flow path to the waste liquid flow path side, thereby completing the collection of the target component.
- This delay time is calculated, for example, by dividing the capacity of the pipe from the flow cell of the detector to the flow path switching unit of the fraction collector by the flow rate of the mobile phase (the amount of liquid fed per unit time) (for example, Patent Documents). 3).
- the target component detected by the detector reaches the flow channel switching unit when the delay time elapses
- the target component is collected by switching the flow channel switching unit.
- the pipe diameter and cross-sectional area have manufacturing errors within tolerances. Since the delay time is calculated based on the pipe capacity determined by the product of the pipe diameter (cross-sectional area) and length, the longer the pipe, the greater the influence of the pipe diameter error, and the delay time becomes inaccurate. Become.
- the target component that has passed through the flow cell flows through the pipe while diffusing in the mobile phase, and reaches the flow path switching unit. Therefore, when reaching the flow path switching unit, the peak start point of the target component is delayed and the peak width is also broad. As a result, there has been a problem in that sorting is started even when the target component has not sufficiently reached, or sorting is terminated even though the peak of the target component continues. These could not be covered by the simple delay time calculated by dividing the pipe capacity by the flow rate.
- the problem to be solved by the present invention is to provide a preparative chromatograph capable of reliably collecting a target component.
- the present invention which has been made to solve the above problems, is a preparative chromatograph for collecting target components in a sample temporally separated in a chromatographic column in each preparative container, a) a detector having a flow cell housed in a housing and a detector for detecting a component passing through the flow cell; b) a first pipe connecting the column and the inlet end of the flow cell; c) a flow path switching unit that selectively accommodates a component that has passed through the flow cell contained in the housing and flows into a sorting flow path or a waste liquid flow path that is a flow path connected to the sorting container; d) It is provided with the 2nd piping which connects the outlet end of the flow cell and the channel change part which were stored in the case.
- the fraction collector (flow channel switching unit and sorting container) and the detection unit are housed in separate housings, and the piping connecting the flow cell and the flow channel switching unit is in these housings. It was arranged to connect. For this reason, depending on the arrangement of both housings, the length of the pipe connecting them becomes long, and the error of the delay time and the diffusion of the components in the pipe become large.
- the detection unit (flow cell) and the flow path switching unit are housed in the same casing, the length of the second pipe is typically made shorter than before. can do.
- the error of piping capacity can be made small, and delay time can be made more accurate than before.
- the target component can be collected more reliably than before.
- an absorptiometer using an LED as a light source can be suitably used.
- a white light source such as a deuterium lamp is used. Therefore, it is necessary to use a spectroscopic unit having a diffraction grating for extracting light of a desired wavelength and a motor for driving the diffraction grating, and it is difficult to accommodate the entire absorption spectrophotometer in the case of the fraction collector.
- a spectroscopic unit diffraction grating and motor
- the entire detector can be reduced in size and accommodated in the housing.
- a rack containing a plurality of preparative containers is usually arranged in the casing. Also, a fractionation head to which the outlet end of the sorting channel is attached, and the sorting head is moved in the horizontal and vertical directions so that the outlet end of the sorting channel is positioned above a predetermined sorting container.
- the drive mechanism is provided.
- a flow cell and a flow path switching unit are mounted on the fractionation head. That is, since the flow cell and the flow path switching unit move together with the fractionation head, it is not necessary to consider the movement of the fractionation head, and the second pipe can be further shortened. Further, if the flow cell and the flow path switching unit are arranged adjacent to each other, the target component can be collected without delay time.
- the preparative liquid chromatograph of the present embodiment is roughly divided into a liquid chromatograph unit 10 for separating a target component contained in a sample, a fraction collector 20 for collecting a target component separated by the liquid chromatograph unit 10, and these components. It is comprised from the control part 30 which controls operation
- the mobile phase in the mobile phase container 11 is sucked by the liquid feed pump 12 and sent to the column 14 at a predetermined flow rate.
- a sample containing the target component is injected from the sample injection unit 13 and transported to the column 14 by the flow of the mobile phase.
- the target component in the sample is temporally separated and eluted in the column 14.
- Each unit of the liquid chromatograph unit 10 is housed in each housing and connected by a pipe.
- the fraction collector 20 includes an absorptiometer 21 using three LEDs 211a, 211b, and 211c having different emission wavelengths as a light source, a fractionation head 22, a moving mechanism (rail 23, motor, etc.) of the fractionation head 22, and an electromagnetic valve 24. I have.
- the absorptiometer 21 and the electromagnetic valve 24 are connected by a second pipe 27 and placed on the upper surface of the fractionation head 22, and move along the rail 23 together with the fractionation head 22.
- a plurality of sorting containers 26 accommodated in a rack 25 are placed on the fraction collector 20. Each part of the fraction collector 20 is accommodated in one housing.
- FIG. 3 shows the main configuration of the absorptiometer 21.
- the three LEDs 211a, 211b, and 211c are light sources that emit light in a wavelength band that is absorbed by the three types of target components to be sorted, and are based on a control signal from the sorting control unit 32 described later.
- the flow cell 212 is irradiated in a time division manner (that is, light emitted from the three LEDs in order).
- the measurement light that has passed through the flow cell 212 is detected by the first photodiode 213.
- a part of the light emitted from each LED 211a, 211b, 211c is detected by the second photodiode 214. Detection signals from the first photodiode 213 and the second photodiode 214 are sent to the control unit 30. In the control unit 30, after the absorbances of light of three types of wavelengths are calculated, a chromatogram is created and displayed on the screen of the display unit 50 described later.
- the solenoid valve 24 is switched by a sorting control unit 32 described later, and the target component that has passed through the flow cell 212 is collected in a sorting container through the sorting channel. After the target component has passed, the solenoid valve 24 is switched again by the sorting control unit 32, and the component that has passed through the flow cell 212 is guided to the waste liquid flow path.
- the control unit 30 includes a storage unit 31 and a sorting control unit 32.
- the fractionation control unit 32 is a functional block that controls the operation of each unit of the liquid chromatograph unit 10 and the fraction collector 20. Further, the input unit 40 and the display unit 50 are connected.
- the sorting control unit 32 displays a sorting condition input screen on the display unit 50, and allows the user to input the piping capacity of the second piping 27 and the feeding flow rate of the feeding pump 12. When these are input, the delay time is calculated from the pipe capacity and the liquid feeding flow rate and stored in the storage unit 31. The delay time is the time required for the (target) component detected by the absorptiometer 21 to reach the electromagnetic valve 24.
- the sorting control unit 32 switches the flow path of the electromagnetic valve 24 to the sorting flow path side when the delay time has elapsed from the start of detection of the target component in the absorptiometer 21 and starts collecting the target component. When the delay time has elapsed from the end of detection of the target component, the flow path of the electromagnetic valve 24 is switched to the waste liquid flow path side, and the sampling of the target component is ended.
- the absorptiometer 21 in this embodiment uses LEDs 211a, 211b, and 211c that emit light having a wavelength that is absorbed by the target component as described above. Therefore, it is not necessary to provide a spectroscopic unit unlike a conventional absorptiometer using a white light source such as a mercury lamp. Therefore, the absorptiometer 21 is small and can be accommodated in the fractionation head 22. Further, in this embodiment, since the internal electromagnetic valve 24 of the fractionation head 22 is also accommodated, the pipe length of the second pipe 27 connecting the flow cell 212 and the electromagnetic valve 24 of the absorptiometer 21 is shorter than before.
- FIG. 2 shows an example in which the absorptiometer 21 and the electromagnetic valve 24 are housed in the fractionation head 22, the absorptiometer 21 and the electromagnetic valve 24 are mounted on the upper surface and side surfaces of the absorptiometer 21. You can also.
- FIG. 4 compares the configurations of the present example and the comparative example. In both the present example and the comparative example, the flow rate was set to 1,000 ⁇ L / min.
- the diameter of the first pipe 15 (column 14 to flow cell 212) of the preparative chromatograph of this example is 0.1 mm, the length is 1000 mm, and the volume is 7.9 ⁇ L.
- the diameter of the second pipe 27 (flow cell 212 to solenoid valve 24) is 0.1 mm, the length is 50 mm, and the capacity is 0.4 ⁇ L.
- the diameter of the first pipe (column to flow cell) is 0.1 mm, the length is 300 mm, and the capacity is 2.4 ⁇ L.
- the diameter of the second pipe (flow cell to solenoid valve) is ⁇ 0.3 mm, the length is 1000 mm, and the capacity is 70.7 ⁇ L.
- the reason why the diameter of the conventional second pipe is different is that the pressure resistance of the flow cell of the detector is low. That is, if a thin and long pipe is connected to the outlet end of the flow cell of the detector, the back pressure becomes too high and liquid leakage occurs. On the other hand, since the second pipe 27 is short in the preparative chromatograph of the present embodiment, there is no fear that an excessive back pressure is applied to the flow cell even if the pipe diameter is small.
- the delay time was calculated by dividing the capacity of the second pipe by the flow rate.
- the comparative example was 4.24 sec, whereas the present example was 0.024 sec. That is, it can be seen that the target component can be reliably collected with substantially no delay time.
- the diffusion capacity from the column to the solenoid valve was determined by the following formula described in Non-Patent Document 1.
- ⁇ v is the diffusion capacity ( ⁇ L)
- d is the pipe diameter (mm)
- L is the pipe length (mm)
- F is the flow rate ( ⁇ L / sec)
- Dm is the diffusion coefficient (0.002mm) 2 / sec, general value).
- the target component diffuses to a peak of 3.46 sec (full width at half maximum), whereas in the present embodiment, it is suppressed to a peak at 1.07 sec (full width at half maximum). Therefore, in the preparative chromatograph of the present example, the target component can be reliably collected without diffusing the target component in the mobile phase.
- the above embodiment is an example, and can be appropriately changed in accordance with the gist of the present invention.
- the light from the three types of LEDs 211a, 211b, and 211c is irradiated to the flow cell 212 in a time division manner in the absorptiometer 21, but the elution order of the target component that absorbs light of each wavelength is known in advance.
- the LEDs may be switched in that order.
- what is necessary is just to change suitably the number of LED to be used.
- a mercury lamp having a narrow spectrum similar to the LED may be used instead of the LED.
- the absorptiometer 21 is used as a detector, but other detectors (fluorescence detector, electrical conductivity detector, differential refractive index detector, etc.) can also be used. A plurality of detectors can be used in combination.
- an absorption spectrophotometer using a white light source can be used as in the prior art. In that case, only the flow cell is placed in the fraction collector's sorting head, and the spectroscopic unit (for example, a diffraction grating) that extracts monochromatic light from the white light emitted from the light source is placed at any position inside or outside the housing of the fraction collector. To do.
- the monochromatic light extracted in the spectroscopic unit may be transported by an optical fiber and irradiated to the flow cell.
- the absorptiometer 21 was mounted on the fractionation head 22 in the said Example, if it is in the housing
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Abstract
Description
a) 筐体内に収容されたフローセルと、該フローセルを通過する成分を検出する検出器とを有する検出部と、
b) 前記カラムと前記フローセルの入口端を接続する第1配管と、
c) 前記筐体内に収容された、前記フローセルを通過した成分を前記分取容器に接続される流路である分取流路又は廃液流路に選択的に流す流路切替部と、
d) 前記筐体内に収容された、前記フローセルの出口端と前記流路切替部を接続する第2配管と
を備えることを特徴とする。
なお、従来同様に重水素ランプ等の光源及び分光部を有する検出器を使用する場合には、フローセルのみを上記筐体内に収容し、光ファイバーを用いて光源からの照射光やフローセルを通過した測定光を輸送するようにすればよい。
e) 前記分取流路の出口端が取り付けられ、前記フローセル及び前記第2配管並びに前記流路切替部が搭載された分画ヘッドと、
f) 前記分取流路の出口端を前記各分取容器間で移動させる駆動機構と
を備えることが好ましい。
本実施例の分取クロマトグラフの要部構成を図1に示す。また、分取クロマトグラフのフラクションコレクター20の要部構成を図2に示す。本実施例の分取液体クロマトグラフは、大別して、試料に含まれる目的成分を分離する液体クロマトグラフ部10、液体クロマトグラフ部10で分離された目的成分を採取するフラクションコレクター20、及びこれらの動作を制御する制御部30から構成されている。
上式(1)において、σvは拡散容量(μL)、dは配管の直径(mm)、Lは配管の長さ(mm)、Fは流量(μL/sec)、Dmは拡散係数(0.002mm2/sec, 一般値)である。
さらに、従来同様に、白色光源を用いる吸光分光光度計を用いることもできる。その場合には、フローセルのみをフラクションコレクターの分取ヘッドに配置し、該光源から発せられる白色光から単色光を取り出す分光部(例えば回折格子)はフラクションコレクターの筐体内外の任意の位置に配置する。そして、分光部において取り出した単色光を光ファイバーにより輸送してフローセルに照射すればよい。
その他、上記実施例では吸光光度計21を分画ヘッド22上に載置したが、フラクションコレクターの筐体内であれば別の位置に配置してもよい。
11…移動相容器
12…送液ポンプ
13…試料注入部
14…カラム
15…第1配管
20…フラクションコレクター
21…フローセル
21…吸光光度計
211a~211c…LED
212…フローセル
213…第1フォトダイオード
214…第2フォトダイオード
22…分画ヘッド
23…レール
24…電磁弁
25…ラック
26…分取容器
27…第2配管
30…制御部
31…記憶部
32…分取制御部
40…入力部
50…表示部
Claims (4)
- クロマトグラフのカラムにおいて時間的に分離された試料中の目的成分を各分取容器に採取する分取クロマトグラフであって、
a) 筐体内に収容されたフローセルと、該フローセルを通過する成分を検出する検出器とを有する検出部と、
b) 前記カラムと前記フローセルの入口端を接続する第1配管と、
c) 前記筐体内に収容された、前記フローセルを通過した成分を前記分取容器に接続される流路である分取流路又は廃液流路に選択的に流す流路切替部と、
d) 前記筐体内に収容された、前記フローセルの出口端と前記流路切替部を接続する第2配管と
を備えることを特徴とする分取クロマトグラフ。 - 前記検出部がLED光源を備えた吸光光度計であることを特徴とする請求項1に記載の分取クロマトグラフ。
- 前記検出部が、前記筐体外に配置された光源から発せられ前記フローセルに照射される照射光を輸送する光ファイバー、を備えることを特徴とする請求項1に記載の分取クロマトグラフ。
- e) 前記分取流路の出口端が取り付けられ、前記フローセル及び前記第2配管並びに前記流路切替部が搭載された分画ヘッドと、
f) 前記分取流路の出口端を前記各分取容器間で移動させる駆動機構と
を備えることを特徴とする請求項1から3のいずれかに記載の分取クロマトグラフ。
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PCT/JP2015/065796 WO2016194108A1 (ja) | 2015-06-01 | 2015-06-01 | 分取クロマトグラフ |
CN201580080553.5A CN107615058B (zh) | 2015-06-01 | 2015-06-01 | 制备色谱仪 |
US15/574,661 US20180136174A1 (en) | 2015-06-01 | 2015-06-01 | Preparative chromatograph |
JP2017521367A JP6394803B2 (ja) | 2015-06-01 | 2015-06-01 | 分取クロマトグラフ |
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Cited By (4)
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CN110325830A (zh) * | 2017-02-23 | 2019-10-11 | 锋翔科技公司 | 用于液相色谱的集成照射检测流动池 |
WO2020179003A1 (ja) | 2019-03-06 | 2020-09-10 | 株式会社島津製作所 | 分取クロマトグラフシステム |
DE102022001352A1 (de) | 2021-04-26 | 2022-10-27 | Shimadzu Corporation | Präparativer Chromatograph und Präparationsverfahren unter Verwendung eines präparativen Chromatographen |
US11953474B2 (en) | 2018-12-13 | 2024-04-09 | Shimadzu Corporation | Preparative chromatograph |
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CN108459120A (zh) * | 2018-01-29 | 2018-08-28 | 天津博纳艾杰尔科技有限公司 | 一种纯化制备色谱系统 |
DE102019205509B3 (de) * | 2019-04-16 | 2020-10-08 | Bruker Biospin Gmbh | Totvolumen-freie Fraktionssammel-Vorrichtung |
IT201900013656A1 (it) * | 2019-08-01 | 2021-02-01 | F Lab S R L | Dispositivo per la produzione di schiuma di un liquido, particolarmente di un liquido alimentare quale latte, caffè o simili. |
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JP5092851B2 (ja) * | 2008-04-04 | 2012-12-05 | 株式会社島津製作所 | 分取液体クロマトグラフシステム |
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JP2020508451A (ja) * | 2017-02-23 | 2020-03-19 | フォセオン テクノロジー, インコーポレイテッドPhoseon Technology, Inc. | 液体クロマトグラフィー用の統合型照明検出フローセル |
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WO2020179003A1 (ja) | 2019-03-06 | 2020-09-10 | 株式会社島津製作所 | 分取クロマトグラフシステム |
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Also Published As
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CN107615058A (zh) | 2018-01-19 |
JPWO2016194108A1 (ja) | 2017-11-02 |
US20180136174A1 (en) | 2018-05-17 |
JP6394803B2 (ja) | 2018-09-26 |
CN107615058B (zh) | 2020-01-17 |
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