JP5814499B2 - 溶液中の物質の濃度の測定方法及び装置 - Google Patents
溶液中の物質の濃度の測定方法及び装置 Download PDFInfo
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- 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
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N21/05—Flow-through cuvettes
-
- 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
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
- G01N21/274—Calibration, base line adjustment, drift correction
-
- 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
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- 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
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0193—Arrangements or apparatus for facilitating the optical investigation the sample being taken from a stream or flow to the measurement cell
-
- 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
- G01N2030/746—Optical detectors detecting along the line of flow, e.g. axial
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- G—PHYSICS
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- G—PHYSICS
- G01—MEASURING; TESTING
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- G01N2201/00—Features of devices classified in G01N21/00
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Description
式中、Aは吸光度であり、EはL・mol-1・cm-1単位のモル吸光係数であり、bはcm単位で定義される試料の光路長であり、cはmol・L-1単位で表される溶液中の化合物の濃度である。
i)光源からバンドパスフィルターを通して波長300nm以下の紫外光を送る段階、
ii)バンドパスフィルターから出た紫外光をビームスプリッターに通す段階、
iii)光の第1の部分をビームスプリッターで分岐して、第1の部分の電磁波を対照検出器で定量して対照値を得る段階、
iv)ビームスプリッターを通過する光の第2の部分を既知の光路長の溶液に照射する段階、
v)溶液を通過した電磁波を試料検出器で定量して試料値を得る段階、及び
vi)試料値と対照値から吸光度(A)を計算する段階
を含んでなり、上記光源が300nm以下の波長の光を放射する発光ダイオードであることを特徴とする方法を提供する。
i)溶液を収容するための既知の光路長のセルであって、300nm以下の波長の光に透明なセル、
ii)光源、
iii)バンドパスフィルター、
iv)光を第1の部分及び第2の部分に分割するためのビームスプリッター、
v)ビームスプリッターで分岐された電磁波を検出するための対照検出器、及び
vi)溶液を通過した電磁波を検出するための試料検出器
を備えており、上記光源が300nm以下の波長の光を放射する発光ダイオードであることを特徴とする装置を提供する。
当業者には明らかであろうが、光路長が未知のフローセル(130)を有する同じ装置を用いると、300nm以下に吸収を有する物質の存在を単に検出することができる。溶液中の物質の濃度の決定又は測定には光路長を知得しておく必要がある(ランベルト・ベール則参照)。
20,120 UV発光ダイオード
22 バンドパス干渉フィルター
24 ビームスプリッター
30,130 フローセル
32,132 入口
34,134 出口
36,38,136,138 窓
40 光検出器
42 対照光検出器
140 試料光検出器
150 コリメータレンズ
155 バンドパスフィルター
160 集光レンズ
170 ビームスプリッター
180 対照光検出器
Claims (12)
- 300nm以下に吸収を有する物質の溶液中での濃度を、前記溶液を収容するための既知の光路長のフローセルで測定する方法であって、前記フローセルが、300nm以下の波長の光に透明な一対の窓を有していて、前記一対の窓が前記フローセルの両端に配置されて前記光路長を規定しており、前記フローセルがさらに前記一対の窓のそれぞれに隣接して前記溶液をフローセルに流すための入口と出口を有しており、当該方法が、
i)溶液中に存在する1種以上の化合物から前記物質をクロマトグラフィーによって分離する段階、
ii)300nm以下の波長の光を放射する発光ダイオードである光源からバンド幅10nm未満のバンドパスフィルターを通して波長300nm以下の紫外光を送る段階、
iii)バンドパスフィルターから出た紫外光をビームスプリッターに通す段階、
iv)光の第1の部分をビームスプリッターで分岐して、第1の部分の電磁波を対照検出器で定量して対照値を得る段階、
v)ビームスプリッターを通過する光の第2の部分を前記光路長の溶液に照射する段階、
vi)溶液を通過した電磁波を試料検出器で定量して試料値を得る段階、及び
vii)試料値と対照値から吸光度(A)を計算し、該吸光度を用いて用量−応答曲線により濃度の測定値を得る段階
を含んでなることを特徴とする方法。 - 前記溶液が細胞抽出液、細胞溶解液及び細胞培養液又はこれらの混合物からなる群から選択される、請求項1記載の方法。
- 既知の内容積のフローセル内の溶液に照射し、該フローセルが300nm以下の波長の光に透明である、請求項1又は請求項2記載の方法。
- 前記対照検出器及び試料検出器がUV感受性光電子増倍管又はUV感受性光ダイオードである、請求項1乃至請求項3のいずれか1項記載の方法。
- 前記物質がタンパク質又はペプチドである、請求項1乃至請求項4のいずれか1項記載の方法。
- 前記物質が核酸である、請求項1乃至請求項4のいずれか1項記載の方法。
- 300nm以下に吸収を有する物質の溶液中での濃度を測定するための装置であって、当該装置が、
i)溶液中に存在する1種以上の化合物から前記物質を分離するためのクロマトグラフィー分離手段、
ii)前記溶液を収容するための既知の光路長のフローセルであって、前記フローセルが、300nm以下の波長の光に透明な一対の窓を有していて、前記一対の窓が前記フローセルの両端に配置されて前記光路長を規定しており、前記フローセルがさらに前記一対の窓のそれぞれに隣接して前記溶液をフローセルに流すための入口と出口を有している、フローセル、
iii)300nm以下の波長の光を放射する発光ダイオードである光源、
iv)バンド幅10nm未満のバンドパスフィルター、
v)光を第1の部分及び第2の部分に分割するためのビームスプリッター、
vi)ビームスプリッターで分岐された電磁波を検出するための対照検出器、
vii)溶液を通過した電磁波を検出するための試料検出器、及び
viii)試料値と対照値から吸光度(A)を計算し、該吸光度を用いて濃度の測定値を得るための用量−応答曲線
を備えていて、バンドパスフィルターが光源とビームスプリッターの間に配置され、光の第1の部分がフローセルを通して送られ、光の第2の部分が対照検出器に送られることを特徴とする装置。 - 前記フローセルが既知の内容積のものである、請求項7記載の装置。
- 前記対照検出器及び試料検出器がUV感受性光ダイオード又はUV感受性光電子増倍管である、請求項7又は請求項8記載の装置。
- さらにコリメータレンズを備える、請求項7乃至請求項9のいずれか1項記載の装置。
- さらに、集光レンズを含む、請求項7乃至請求項10のいずれか1項記載の装置。
- タンパク質、ペプチド及び核酸からなる群から選択される物質の濃度を測定するための請求項7乃至請求項11のいずれか1項記載の装置の使用。
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GB0524225.0 | 2005-11-29 | ||
GBGB0524225.0A GB0524225D0 (en) | 2005-11-29 | 2005-11-29 | Methods and apparatus for detecting and measuring the concentration of a substance in a solution |
PCT/EP2006/011394 WO2007062800A1 (en) | 2005-11-29 | 2006-11-28 | Methods and apparatus for measuring the concentration of a substance in a solution |
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JP2009517641A JP2009517641A (ja) | 2009-04-30 |
JP2009517641A5 JP2009517641A5 (ja) | 2010-01-21 |
JP5814499B2 true JP5814499B2 (ja) | 2015-11-17 |
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US (1) | US8068227B2 (ja) |
EP (1) | EP1955047B1 (ja) |
JP (1) | JP5814499B2 (ja) |
CA (1) | CA2631369C (ja) |
GB (1) | GB0524225D0 (ja) |
WO (1) | WO2007062800A1 (ja) |
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JP2009168478A (ja) * | 2008-01-11 | 2009-07-30 | Hitachi High-Technologies Corp | 光度計 |
EP2350612B1 (en) * | 2008-11-24 | 2023-05-24 | Cytiva Sweden AB | Flow cell optical detection system |
JP5239000B2 (ja) * | 2009-10-27 | 2013-07-17 | 有限会社光電鍍工業所 | オゾン濃度測定装置 |
DE102010015083B4 (de) * | 2010-04-15 | 2012-04-12 | Eads Deutschland Gmbh | Sensor und Verfahren zur Online-Überwachung der Säurezahl eines Hydraulikfluids in einem Hydrauliksystem in einem Luftfahrzeug |
EP2577270A1 (en) | 2010-05-27 | 2013-04-10 | Tallinn University Of Technology | Method and device for measuring and monitoring concentration of substances in a biological fluid |
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GB0524225D0 (en) | 2006-01-04 |
US20080304048A1 (en) | 2008-12-11 |
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US8068227B2 (en) | 2011-11-29 |
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