Method and apparatus for analyzing samples and collecting sample fractions
Abstract
A method for analyzing a sample using at least one detector and a device for that purpose are disclosed. [Selection diagram] Fig. 1
Term
Projected expiry 4 December 2028.
- Priority
- Filed
- Published
- Today
- Projected expiry
134 claims: 23 independent, 111 dependent
- 1サンプルを分析する方法であって、前記方法は、 (a)液体クロマトグラフィーシステム内の2つまたはそれ以上の検出器から複合信号を発生させるステップを有し、前記複合信号は各検出器からの検出応答成分を有し、 (b)前記方法は、前記複合信号の変化に応じて、画分収集器内に新しいサンプル画分を収集するステップを有する、方法。
- 2請求項1に記載の方法であって、前記複合信号を発生させるステップは、 (a)(i)検出器応答値、(ii)時間の関数として与えられた検出器応答の傾斜(すなわち、与えられた検出器応答の1次微分)、(iii)時間の関数として与えられた検出器応答の傾斜の変化(すなわち、与えられた検出器応答の2次微分)、または(iv)各検出器からの(i)から(iii)までの任意の組み合わせ、を数学的に相関させるステップを有する、方法。
- 3請求項1に記載の方法であって、前記複合信号は、(i)与えられた時間における各検出器の検出器応答時間の積、(ii)与えられた時間における検出器応答の1次微分の積、(iii)与えられた時間における検出器応答の2次微分の積、または(iv)(i)から(iii)までの任意の組み合わせ、を有する、方法。
- 4請求項1に記載の方法であって、前記方法はさらに、 (a)2つまたはそれ以上の光学波長でサンプルを観察し、2つまたはそれ以上の光学波長における2つまたはそれ以上の検出器応答を生成するために、少なくとも1つの検出器を用いるステップを有し、前記複合信号は、2つまたはそれ以上の光学波長における2つまたはそれ以上の検出器応答からの、検出応答成分を有する、方法。
- 5請求項4に記載の方法であって、2つまたはそれ以上の光学波長を観察する少なくとも1つの検出器は、少なくとも1つのUV検出器を有し、前記複合信号は、(i)前記UV検出器の2つまたはそれ以上の光学波長における2つまたはそれ以上の検出器応答からの検出応答成分、および(ii)蒸発性光散乱検出器(ELSD)からの検出応答成分、を有する、方法。
- 6請求項1に記載の方法であって、複合信号は、(i)少なくとも1つのUV検出器からの検出応答成分、および(ii)少なくとも1つの蒸発性光散乱検出器(ELSD)からの検出応答成分、を有する方法。
- 7請求項1に記載の方法であって、さらに (a)少なくとも1つの検出器と流体連通するように位置決めされる、(i)スプリッタポンプ、(ii)シャトルバルブ、または(iii)(i)と(ii)との両方、を介して、液体クロマトグラフィーシステム内の前記少なくとも1つの検出器へ流体流れを能動的に制御するステップ、を有する方法。
- 8請求項1に記載の方法であって、さらに 少なくとも1つの検出器と流体連通するように位置決めされるシャトルバルブを介して、液体クロマトグラフィーシステム内の少なくとも1つの検出器への流体流れを能動的に制御するステップ、を有する方法。
- 9請求項8に記載の方法であって、前記少なくとも1つの検出器は、蒸発性光散乱検出器(ELSD)を有する、方法。
- 10請求項8に記載の方法であって、さらに、 (a)前記シャトルバルブから前記少なくとも1つの検出器へサンプルアリコットを輸送するために、空気を用いるステップを有する、方法。
- 11請求項8に記載の方法であって、前記シャトルバルブは、前記少なくとも1つの検出器へ輸送するために、10秒ごとに少なくとも1サンプルアリコットのサンプリング周波数で、サンプルからサンプルアリコットを取り除く、方法。
- 12サンプルを分析する方法であって、前記方法は、 (a)2つまたはそれ以上の光学波長でサンプルを観察するために、少なくとも1つの検出器を用いるステップと、 (b)(i)第1波長における検出器応答の変化、(ii)第2波長における検出器応答の変化、または(iii)前記第1波長および前記第2波長における検出器応答により表現される複合応答の変化、に応じて、画分収集器に新しいサンプル画分を収集するステップと、を有する、方法。
- 13請求項12に記載の方法であって、さらに、 (a)(i)第1波長における検出器応答の変化、(ii)第2波長における検出器応答の変化、および(iii)前記第1波長および前記第2波長における検出器応答により表現される複合応答の変化、に応じて、前記画分収集器に新しい画分を収集するステップ、を有する、方法。
- 14請求項12に記載の方法であって、さらに、 (a)吸収スペクトルの範囲にわたってn個の波長でサンプルを観察するために、少なくとも1つの検出器内のn個のセンサを用いるステップを有し、nは1より大きい整数であり、 (b)前記方法は、(i)前記n個のセンサからのn個の検出器応答のうちのいずれか1つの変化、または(ii)前記n個の検出器応答により表現される複合応答の変化、に応じて、前記サンプル画分収集器内に新しいサンプル画分を収集するステップを、有する、方法。
- 15請求項14に記載の方法であって、前記吸収スペクトルの範囲は、紫外(UV)領域である、方法。
- 16請求項14に記載の方法であって、前記システムは、n個のセンサだけを有するまたは1つ以上の追加の検出器と組み合わされた、単一のUV検出器を有する、方法。
- 17請求項14に記載の方法であって、さらに、 (a)少なくとも1つの検出器と流体連通するように位置決めされる、(i)スプリッタポンプ、(ii)シャトルバルブ、または(iii)(i)および(ii)の両方、を介して、液体クロマトグラフィーシステム内の少なくとも1つの検出器へ流体流れを能動的に制御するステップ、を有する、方法。
- 18請求項14に記載の方法であって、さらに、 (a)少なくとも1つの検出器に流体連通するように位置決めされるスプリッタポンプを介して、液体クロマトグラフィーシステム内の少なくとも1つの検出器へ流体流れを能動的に制御するステップ、を有する、方法。
- 19サンプルを分析する方法であって、前記方法は、 (a)(i)クロマトグラフィーカラム、(ii)画分収集器、および(iii)第1検出器、を有する液体クロマトグラフィーシステムを提供するステップと、 (b)(iv)前記第1検出器と流体連通するように位置決めされる、スプリッタポンプまたはシャトルバルブを介して、前記第1検出器への流体流れを能動的に制御するステップと、を有する、方法。
- 20請求項19に記載の方法であって、前記第1検出器へ流体流れを能動的に制御するステップは、 (a)(i)前記スプリッタポンプまたは前記シャトルバルブを駆動し、(ii)前記スプリッタポンプまたは前記シャトルバルブを停止し、(iii)前記スプリッタポンプまたは前記シャトルバルブの1つまたはそれ以上の流量または圧力の設定を変更し、または(iv)(i)から(iii)までの任意の組み合わせを行うために、前記スプリッタポンプまたは前記シャトルバルブに駆動信号を送るステップ、を有する、方法。
- 21請求項20に記載の方法であって、1つまたはそれ以上の流量または圧力の設定は、(i)バルブ位置、(ii)スプリッタポンプまたはシャトルバルブの圧力、(iii)バルブへの空気圧、または(iv)(i)から(iii)までの任意の組み合わせ、を有する、方法。
- 22請求項20に記載の方法であって、前記駆動信号は、電気信号、気体信号、ディジタル信号、または無線信号を有する、方法。
- 23請求項19に記載の方法であって、前記第1検出器への流体流れを能動的に制御するステップは、前記スプリッタポンプを介して、前記第1検出器へ流体をポンプ輸送するステップを有する、方法。
- 24請求項19に記載の方法であって、前記第1検出器への流体流れを能動的に制御するステップは、前記スプリッタポンプを介して、前記第1検出器から流体を汲み上げるステップを有する、方法。
- 25請求項19に記載の方法であって、前記第1検出器への流体流れを能動的に制御するステップは、シャトルバルブを介して前記第1検出器へ1つまたはそれ以上のサンプルアリコットを提供するステップを有する、方法。
- 26請求項19に記載の方法であって、前記液体クロマトグラフィーシステムはさらに、(v)第2検出器を有し、前記第1検出器への流体流れを能動的に制御するステップは、前記第1検出器と前記第2検出器との間で流体流れを分割するステップを有する、方法。
- 27請求項19に記載の方法であって、前記液体クロマトグラフィーシステムは、さらに、(v)第2検出器を有し、前記方法はさらに、 (a)前記第検出器がサンプル内の成分を検出することに応じて、前記スプリッタポンプまたは前記シャトルバルブに、前記スプリッタポンプまたは前記シャトルバルブが前記第1検出器への流体流れを開始または停止させるように指示する信号を送るステップ、を有する、方法。
- 28請求項19に記載の方法であって、前記液体クロマトグラフィーシステムはさらに(v)第2検出器、および(vi)第2スプリッタポンプまたは第2シャトルバルブを有し、前記方法はさらに、 (a)前記第2スプリッタポンプまたは前記第2シャトルバルブを介して、前記第2検出器への流体流れを能動的に制御するステップ、を有する、方法。
- 29請求項19に記載の方法であって、さらに、 (a)前記第1検出器からの1つまたはそれ以上の信号に応じて、1つまたはそれ以上のサンプル画分を収集するステップを有する、方法。
- 30請求項26に記載の方法であって、さらに、 (a)(i)前記第1検出器、(ii)前記第2検出器、または(iii)前記第1検出器および前記第2検出器の両方、からの1つまたはそれ以上の検出器信号に応じて、1つまたはそれ以上のサンプル画分を収集するステップ、を有する方法。
- 31請求項26に記載の方法であって、前記液体クロマトグラフィーシステムは、1つまたはそれ以上の非破壊式検出器を備えて破壊式の検出器を備えない、非破壊式システムである、方法。
- 32請求項19に記載の方法であって、さらに、 (a)少なくとも1つの検出器から検出器信号を発生させるステップを有し、前記検出器信号は、(i)時間の関数としての検出器応答の傾斜(すなわち、検出器応答の1次微分)、(ii)時間の関数としての検出器応答の傾斜の変化(すなわち、検出器応答の2次微分)、(iii)随意選択として、検出器応答値の閾値、または(iv)少なくとも(i)または(ii)を含む(i)から(iii)までの任意の組み合わせ、に応答して発生させられる、方法。
- 33請求項32に記載の方法であって、前記少なくとも1つの検出器から検出器信号を発生させるステップは、 (a)(i)時間に関する傾斜データ、(ii)時間に関する傾斜データの変化、(iii)随意選択として、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、の実際の変化を、検出器応答において起こり得るノイズから識別するために、(i)時間に関する傾斜データ、(ii)時間に関する傾斜データの変化、(iii)随意選択として、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、をローパス数値フィルタ処理するステップを有する、方法。
- 34請求項26に記載の方法であって、さらに、 (a)各検出器からの検出器応答成分を有する複合信号を発生させるステップと、 (b)前記複合信号に応じて、新しいサンプル画分を収集するステップと、を有する、方法。
- 35請求項34に記載の方法であって、前記複合信号を発生させるステップは、 (a)(i)検出器応答値、(ii)時間の関数として与えられた検出器応答の傾斜(すなわち、与えられた検出器応答の1次微分)、(iii)時間の関数としてい与えられた検出器応答の傾斜の変化(すなわち、与えられた検出器応答の2次微分)、または(iii)(i)から(iii)までの任意の組み合わせ、を数学的に相関させるステップを有する、方法。
- 36請求項35に記載の方法であって、前記複合信号は、(i)与えられた時間における各検出器の検出器応答値の積、(ii)与えられた時間における検出器応答の1次微分の積、(iii)与えられた時間における検出器応答の2次微分の積、(iv)(i)から(iii)までの任意の組み合わせ、を有する、方法。
- 37請求項19に記載の方法であって、前記液体クロマトグラフィーシステムは、(i)検出器応答の検出と(ii)前記検出応答から発生させられる信号に基づいてサンプル画分を収集するステップとの間の最大時間遅れは、約2.0秒より小さい、方法。
- 38サンプルを分析する方法であって、前記方法は、 (a)(i)クロマトグラフィーカラム、(ii)システム内に破壊式検出器を備えない2つまたはそれ以上の非破壊式検出器、及び(iii)前記2つまたはそれ以上の非破壊式検出器と流体連通する画分収集器、を有する、非破壊式システムの液体クロマトグラフィーシステムを提供するステップと、 (b)前記2つまたはそれ以上の非破壊式検出器からの検出器信号に応じて、1つまたはそれ以上のサンプル画分を収集するステップと、を有する方法。
- 39請求項38に記載の方法であて、さらに、 (a)各検出器からの検出応答成分を有する複合信号を発生させるステップと、 (b)前記複合信号の変化に応じて、新しいサンプル画分を収集するステップと、を有する方法。
- 40請求項38に記載の方法であて、さらに、 (a)少なくとも1つの非破壊式検出器と流体連通するように位置決めされるスプリッタポンプを介して、少なくとも1つの非破壊式検出器への流体流れを能動的に制御するステップ、を有する方法。
- 41サンプルを分析する方法であって、前記方法は、 (a)液体クロマトグラフィーシステム内の少なくとも1つの検出器から検出器信号を発生させるステップを有し、前記検出器信号は、(i)時間の関数と指定の検出器応答の傾斜(すなわち検出器応答の1次微分)、(ii)時間の関数としての検出器応答の傾斜の変化(すなわち検出器応答の2次微分)、(iii)随意選択として、検出器応答値の閾値、または(iv)少なくとも(i)または少なくとも(ii)を含む、(i)から(iii)までの任意の組み合わせ、に応じて発生させられ、 (b)前記方法は、前記少なくとも1つの検出器からの少なくとも1つの検出器信号に応じて、1つまたはそれ以上のサンプル画分を収集するステップ、を有する方法。
- 42請求項41に記載の方法であって、前記少なくとも1つの検出器から検出器信号を発生させるステップは、さらに、 (a)(i)時間に関する傾斜データ、(ii)時間に関する傾斜データの変化、(iii)随意選択として、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、の実際の変化を、検出器応答中の発生しうるノイズから識別するために、(i)時間に関する傾斜データ、(ii)時間に関する傾斜データの変化、(iii)随意選択として、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、を数値フィルタリング処理するステップ、を有する方法。
- 43請求項41に記載の方法であって、前記液体クロマトグラフィーシステムは、2つまたはそれ以上の検出器を有する、方法。
- 44請求項43に記載の方法であって、さらに、 (a)各検出器からの検出応答成分を含む複合信号を発生させるステップと、 (b)前記複合信号の変化に応じて、新しいサンプル画分を収集するステップと、を有する方法。
- 45請求項41に記載の方法であって、さらに、 (a)少なくとも1つの検出器と流体連通するように位置決めされる、(i)スプリッタポンプおよび(ii)シャトルバルブの少なくとも一方を介して、液体クロマトグラフィーシステム内の少なくとも1つの検出器への流体流れを能動的に制御するステップを有する、方法。
- 46サンプルを分析する方法であって、前記方法は、 (a)液体クロマトグラフィーシステムの画分収集器内にサンプル画分を収集するステップを有し、前記画分収集器は、少なくとも1つの検出器からの1つまたはそれ以上の信号を認識し、受信し、および処理して、前記1つまたはそれ以上の信号に基づいて1つまたはそれ以上のサンプル画分を収集するように構成される、方法。
- 47請求項1乃至46のいずれか一項に記載の方法であって、液体クロマトグラフィーシステムは、少なくとも1つのUV検出器、少なくとも1つの蒸発性光散乱検出器(ELSD)、少なくとも1つの質量スペクトル計、少なくとも1つの凝集核形成光散乱検出器(CNLSD)、少なくとも1つのコロナ放電検出器、少なくとも1つの屈折率検出器(RID)、少なくとも1つの蛍光検出器(FD)、少なくとも1つのカイラル検出器(CD)、およびこれらの任意の組み合わせ、から選択される少なくとも1つの検出器を有する、方法。
- 48請求項1乃至47のいずれか一項に記載の方法のステップの1つまたはそれ以上を実行するための、コンピュータが実行可能な命令が格納された、コンピュータで読取り可能な媒体。
- 49請求項1乃至47のいずれか一項に記載の方法を用いてサンプルを分析することができる装置。
- 50請求項1乃至47のいずれか一項に記載の方法を用いるサンプルの分析に貢献することができる、装置または装置コンポーネント。
- 51請求項48に記載のコンピュータで読取り可能な媒体を用いて、サンプルの分析に貢献することができる装置または装置コンポーネント。
- 52サンプルを分析する装置であって、前記装置は、 (a)液体クロマトグラフィーシステム内の2つまたはそれ以上の検出器から複合信号を発生させるように構成されたシステムハードウェアを有し、前記複合信号は、各検出器からの検出応答成分を有し、 (b)前記装置は、前記複合信号の変化に応じて、新しいサンプル画分を収集するように構成される画分収集器、を有する、装置。
- 53請求項52に記載の装置であって、前記複合信号は、(i)検出器応答値、(ii)時間の関数として与えられた検出器応答の傾斜(すなわち与えられた検出器応答の1次微分)、(iii)時間の関数として与えられた検出器応答の傾斜の変化(すなわち与えられた検出器応答の2次微分)、または(iv)各検出器からの(i)から(iii)までの任意の組み合わせ、の間の数学的な相関を有する、装置。
- 54請求項52に記載の装置であって、前記複合信号は、(i)与えられた時間における各検出器の検出器応答値の積、(ii)与えられた時間における検出器応答の1次微分の積、(iii)与えられた時間における検出器応答の2次微分の積、または(iv)(i)から(iii)までの任意の組み合わせ、を有する、装置。
- 55請求項52に記載の装置であって、前記装置は、2つまたはそれ以上の波長でサンプルを観察するように構成される2つまたはそれ以上の検出器と、(i)第1波長における検出器応答の変化、(ii)第2波長における検出器応答の変化、または(iii)前記第1波長および前記第2波長における検出器応答により表現される複合応答の変化、に応じて前記サンプル画分収集器が新しいサンプル画分を収集することを可能にするシステムハードウェハと、を有する方法。
- 56請求項52に記載の装置であって、前記装置はさらに、少なくとも1つの検出器に流体連通するように位置決めされ且つ少なくとも1つの検出器への流体流れを能動的に制御する、スプリッタポンプまたはシャトルバルブ、を有する、装置。
- 57請求項52に記載の装置であって、前記装置はさらに、少なくとも1つの検出器に流体連通するように位置決めされ且つ少なくとも1つの検出器への流体流れを能動的に制御する、シャトルバルブ、を有する、装置。
- 58請求項57に記載の装置であって、前記少なくとも1つの検出器は、蒸発性光散乱検出器(ELSD)を有する、装置。
- 59請求項52に記載の装置であって、前記装置はさらに、ガスまたは液体供給部を有し、前記ガスまたは液体供給部は、シャトルバルブから少なくとも1つの検出器へサンプルアリコットを輸送するように構成される、装置。
- 60請求項59に記載の装置であって、前記シャトルバルブは、10秒毎に少なくとも1サンプルのサンプリング周波数で、少なくとも1つの検出器へ輸送するために、サンプルからサンプルアリコットを取り除くようにプログラムされる、装置。
- 61サンプルを分析する装置であって、前記装置は、 (a)2つまたはそれ以上の光学波長でサンプルを観察するように構成される少なくとも1つの検出器と、 (b)(i)第1波長における検出器応答の変化、(ii)第2波長における検出器応答の変化、または(iii)前記第1波長および前記第2波長で表現される複合応答の変化、に応じて、新しいサンプルを収集するように構成されるサンプル画分収集器と、を有する装置。
- 62請求項61に記載の装置であって、前記画分収集器は、(i)第1波長における検出器応答の変化、(ii)第2波長における検出器応答の変化、および(iii)前記第1波長および前記第2波長で表現される複合応答の変化、に応じて、新しいサンプルを収集するように構成される、装置。
- 63請求項61に記載の装置であって、前記少なくとも1つの検出器は、吸収スペクトルの範囲にわたるn個の波長でサンプルを観察するように構成されるn個のセンサを有し、ここでnは1より大きい整数であり、前記画分収集器は、(i)前記n個のセンサからの検出器応答の任意の1つの変化、または(ii)n個の検出器応答により表現される複合応答の変化、に応じて、新しいサンプルを収集するように構成される、装置。
- 64請求項63に記載の装置であって、前記吸収スペクトルの範囲は紫外(UV)領域である、装置。
- 65請求項63に記載の装置であって、前記システムは、n個のセンサだけを備えるまたは1つまたはそれ以上の追加の検出器との組み合わせた、単一のUV検出器を有する、装置。
- 66請求項61に記載の装置であって、さらに、液体クロマトグラフィーシステム内の少なくとも1つの検出器への流体流れを能動的に制御するために、(i)スプリッタポンプおよび(ii)シャトルバルブの少なくとも一方を有する、装置。
- 67請求項66に記載の装置であって、液体クロマトグラフィーシステム内の少なくとも1つの検出器への流体流れを能動的に制御するために、少なくとも1つのシャトルバルブを有する、装置。
- 68請求項67に記載の装置であって、前記少なくとも1つの検出器は蒸発性光散乱検出器(ELSD)を有する、装置。
- 69サンプルを分析するための装置であって、前記装置は、 (a)クロマトグラフィーカラムと、 (b)画分収集器と、 (c)第1検出器と、 (d)前記第1検出器と流体連通するように位置決めされるスプリッタポンプまたはシャトルバルブと、を有し、前記スプリッタポンプまたは前記シャトルバルブは、前記第1検出器への流体流れを能動的に制御するように構成される、装置。
- 70請求項69に記載の装置であって、さらに、前記第1検出器が前記スプリッタポンプまたは前記シャトルバルブに駆動信号を送り、(i)前記スプリッタポンプまたは前記シャトルバルブを駆動、(ii)前記スプリッタポンプまたは前記シャトルバルブを停止し、(iii)前記スプリッタポンプまたは前記シャトルバルブの1つまたはそれ以上の流量または圧力の設定を変更し、または(iv)(i)から(iii)までの任意の組み合わせを行う、ことを可能にする、システムハードウェアを有する、装置。
- 71請求項70に記載の装置であって、前記1つまたはそれ以上の流量または圧力の設定は、(i)バルブ位置、(ii)スプリッタポンプまたはシャトルバルブの圧力、(iii)バルブへの空気圧力、または(iv)(i)から(iii)までの任意の組み合わせ、を有する、装置。
- 72請求項70に記載の装置であって、駆動信号は、電気信号、気体信号、ディジタル信号、または無線信号、を有する、装置。
- 73請求項69に記載の装置であって、前記スプリッタポンプは、T部と第1検出器との間に位置決めされる、装置。
- 74請求項69に記載の装置であって、前記第1検出器は、T部と前記スプリッタポンプとの間に位置決めされる、装置。
- 75請求項69に記載の装置であって、さらに、 (e)第2検出器、を有する装置。
- 76請求項75に記載の装置であって、前記スプリッタポンプまたは前記シャトルバルブは、前記第1検出器と前記第2検出器との間で流体流れの容積を分割する、装置。
- 77請求項75に記載の装置であって、サンプル中の検出成分に応じて、第2検出器は、前記スプリッタポンプまたは前記シャトルバルブに、前記スプリッタポンプまたは前記シャトルバルブが前記第1検出器への流体流れを開始または停止させる信号を送るように構成される、装置。
- 78請求項69に記載の装置であって、さらに、 (f)第2T部の第2出口に流体連通する第2検出器と、 (g)(vi)第2スプリッタポンプまたは第2シャトルバルブと、を有し、前記第2スプリッタポンプまたは前記第2シャトルバルブは、前記第2検出器への流体流れを能動的に制御する、装置。
- 79請求項69に記載の装置であって、画分収集器は、第1検出器からの1つまたはそれ以上の信号に応じて、1つまたはそれ以上のサンプル画分を収集するように構成される、装置。
- 80請求項75に記載の装置であって、画分収集器は、(i)第1検出器、(ii)第2検出器、または(iii)第1検出器と第2検出器との両方、からの1つまたはそれ以上の検出器信号に応じて、1つまたはそれ以上のサンプル画分を収集する、装置。
- 81請求項75に記載の装置であって、前記装置は、1つまたはそれ以上の非破壊式検出器を備えシステム内に破壊式検出器を備えない、非破壊式システムである、装置。
- 82請求項69に記載の装置であって、さらに、システムハードウェアを有し、前記システムハードウェアは、少なくとも1つの検出器からの検出器信号の発生を可能にし、前記検出器信号は、(i)時間の関数としての検出器応答の傾斜(すなわち、検出器応答の1次微分)、(ii)時間の関数と指定の検出器応答の傾斜の変化(すなわち、検出応答の2次微分)、(iii)随意選択として、検出器応答値の閾値、または(iv)少なくとも(i)または少なくとも(ii)を有する(i)から(iii)までの任意の組み合わせ、に応じて生成される、装置。
- 83請求項82に記載の装置であって、前記システムハードウェハは、(i)時間に関する傾斜データ、(ii)時間に関する傾斜の変化、(iii)随意選択として、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、の実際の変化を、検出器応答内の存在し得るノイズから識別するために、(i)時間に関する傾斜データ、(ii)時間に関する傾斜の変化、(iii)随意選択として、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、をローパス数値フィルタリング処理することを可能にする、装置。
- 84請求項75に記載の装置であって、さらにシステムハードウェアは、各検出器からの検出器応答成分を有する複合信号の発生を可能にする、前記システムハードウェアは、前記複合信号の変化に応じて、画分収集器に新しいサンプル画分を収集させる命令を送るように構成される、装置。
- 85請求項85に記載の装置であって、前記複合信号は、(i)検出器応答値、(ii)時間の関数として与えられた検出器応答の傾斜(すなわち与えられた検出応答の1次微分)、(iii)時間の関数として与えられた検出応答の傾斜の変化(すなわち与えられた検出応答の2次微分)、または(iv)(i)から(iii)までの任意の組み合わせ、との間に数学的な相関を有する、装置。
- 86請求項84に記載の装置であって、前記複合信号は、(i)与えられた時間における各検出器の検出器応答値の積、(ii)与えられた時間における検出器応答の1次微分の積、(iii)与えられた時間における検出器応答の2次微分の積、または(iv)(i)から(iii)までの任意の組み合わせ、を有する装置。
- 87請求項69に記載の装置であって、前記装置は、2つまたはそれ以上の光学波長でサンプルを観察するように構成される少なくとも1つの検出器を有し、前記装置は、システムハードウェアを有し、前記システムハードウェアは、(i)第1波長における検出器応答の変化、(ii)第2波長における検出応答の変化、または(iii)前記第1波長および前記第2波長により表現される複合検出応答の変化、に応じて、前記画分収集器が、新しいサンプル画分を収集することを可能にする、装置。
- 88請求項69に記載の装置であって、前記装置は、吸収スペクトルの範囲にわたってm個の波長でサンプルを観察するように構成されるn個のセンサを備える単一の検出器を有し、nは1より大きい整数であり、前記装置は、前記画分収集器が(i)n個のUV波長におけるn個の検出器応答の任意の1つの変化、(ii)n個の検出器応答により表現される複合信号の変化、に応じて新しいサンプル画分を収集することができるようにするシステムハードウェアを有する、装置。
- 89請求項69に記載の装置であって、前記画分収集器は、少なくとも1つの検出器からの1つまたはそれ以上の信号を認識し、受信し、処理して、前記1つまたはそれ以上の信号に基づいて1つまたはそれ以上のサンプル画分を収集するように構成される、装置。
- 90請求項70に記載の装置であって、前記装置は、(i)検出器応答の検出と(ii)前記検出器応答から生成される信号に基づくサンプル画分の収集のステップとの間の最大時間遅れが、約2.0秒未満である、装置。
- 91サンプルを分析するための装置であって、前記装置は、 (a)液体クロマトグラフィーシステム内の少なくとも1つの検出器からの検出器信号の生成を可能にするシステムハードウェアを有し、前記検出器信号は、(i)時間の関数としての検出器応答の傾斜(すなわち、検出器応答の1次微分)、(ii)時間の関数としての検出器応答の傾斜の変化(すなわち、検出器応答の2次微分)、または(iii)少なくとも(i)または少なくとも(ii)を含む(i)から(iii)までの任意の組み合わせ、に応じて、発生させられる、装置。
- 92請求項91に記載の装置であて、前記システムハードウェアは、(i)時間に関する傾斜データ、(ii)時間に関する傾斜データの変化、(iii)随意選択で、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、を前記検出器応答の存在し得るノイズから識別するために、(i)時間に関する傾斜データ、(ii)時間に関する傾斜データの変化、(iii)随意選択で、検出器応答値の閾値、または(iv)(i)から(iii)までの任意の組み合わせ、をローパス数値フィルタリング処理をする、装置。
- 93請求項91に記載の装置であって、前記装置はさらに、少なくとも1つの検出器からの検出器信号に応じて1つまたはそれ以上のサンプル画分を収集するように構成される画分収集器を有する、装置。
- 94請求項91に記載の装置であって、前記装置は2つまたはそれ以上の検出器を有する、装置。
- 95請求項94に記載の装置であって、前記システムハードウェアは、各検出器からの検出器応答を有する複合信号を生成することができる、装置。
- 96請求項95に記載の装置であって、前記複合信号は、(i)検出器応答値、(ii)時間の関数として与えられた検出器応答の傾斜(すなわち、与えられた検出器応答の1次微分)、(iii)時間の関数として与えられた検出器応答の傾斜の変化(すなわち、与えられた検出器応答の2次微分)、または(iv)各検出器からの(i)から(iii)までに任意の組み合わせ、の間に数学的な相関がある、装置。
- 97請求項95に記載の装置であって、前記複合信号は、(i)与えられた時間における各検出器の検出器応答値の積、(ii)与えられた時間における検出器応答値の1次微分の積、(iii)与えられた時間における検出器応答の2次微分の積、または(iv)(i)から(iii)までの任意の組み合わせ、を有する、装置。
- 98請求項95に記載の装置であって、前記装置はさらに、前記複合信号の変化におうじて新しいサンプル画分を収集する画分収集器を有する、装置。
- 99請求項91に記載の装置であって、前記装置は、(i)クロマトグラフィーカラムと、(ii)2つまたはそれ以上の非破壊式検出器とを有し、前記装置は、システム内に破壊式検出器を含まず、前記装置は、(iii)2つまたはそれ以上の非破壊式検出器と流体連通する画分収集器を有する、装置。
- 100請求項94に記載の装置であって、前記装置は、少なくとも1つの検出器と流体連通するように位置決めされ、且つ、少なくとも1つの検出器への流体流れを能動的に制御する、スプリッタポンプまたはシャトルバルブを有する、装置。
- 101サンプルを分析するための装置であって、前記装置は、 (a)液体クロマトグラフィーシステム内の画分収集器を有し、前記画分収集器は、少なくとも1つの検出器からの1つまたはそれ以上の信号を認識し、受信し、処理することができ、また、1つまたはそれ以上の信号に基づいて1つまたはそれ以上のサンプル画分を収集するように構成される、装置。
- 102請求項52乃至101のいずれか一項に記載の装置であって、前記装置は、少なくとも1つのUV検出器、少なくとも1つの蒸発性光散乱検出器(ELSD)、少なくとも1つの質量スペクトル計(MS)、少なくとも1つの凝集核形成光散乱検出器(CNLSD)、少なくとも1つのコロナ放電検出器(CDD)、少なくとも1つの屈折率検出器(RID)、少なくとも1つの蛍光検出器(FD)、少なくとも1つのカイラル検出器(CD)、またはこれらの任意の組み合わせ、から選択される少なくとも1つの検出器を有する、装置。
- 103クロマトグラフィーを用いてサンプルを分析する方法であって、前記方法は、 (a)少なくとも1つの非発色性の検出対象物質を有するサンプルを観察するための少なくとも1つの検出器を用いるステップと、 (b)前記非発色性物質の検出器応答の変化に応じて、新しいサンプル画分を画分収集器内に収集するステップと、を有する方法。
- 104請求項103に記載の方法であって、前記サンプルは非発色性の可動フェーズを有する、方法。
- 105クロマトグラフィーを用いて流体サンプルを分析するための装置であって、前記装置は、 (a)サンプル中の発色性物質および非発色性物質を検出することができる少なくとも1つの検出器と、 (b)前記非発色性物質の検出器応答の変化に応答することができる画分収集器と、を有する装置。
- 106請求項105に記載の装置であって、前記サンプルは非発色性の可動フェーズを有する、装置。
- 107クロマトグラフィーを用いて流体サンプルを分析する方法であって、前記方法は、 (a)第1流体を提供するステップと、 (b)前記第1流体の流れ特性に実質的に影響を与えることなく、前記第1流体から流体のアリコットサンプルを取り除くためにシャトルバルブを用いるステップと、 (c)流体の前記アリコットサンプルを観察するために少なくとも1つの検出器を用いるステップと、 (d)検出器応答の変化に応じて、前記第1流体からの新しいサンプル画分を画分収集器に収集するステップと、を有する方法。
- 108請求項107に記載の方法であって、前記シャトルバルブを通る前記第1流体の流れは実質的に層流である、方法。
- 109請求項107に記載の方法であって、前記シャトルバルブを通る前記第1流体の圧力は、実質的に増加せず、または実質的に一定である、方法。
- 110請求項107に記載の方法であって、前記方法はさらに、流体のアリコットサンプルを前記検出器へ運ぶ第2流体を有する、方法。
- 111請求項107に記載の方法であって、前記シャトルバルブを通る前記第2流体の流れは実施的に層流である、方法。
- 112請求項107に記載の方法であって、前記シャトルバルブを通る前記第2流体の圧力は、実質的に増加せず、または実質的に一定である、方法。
- 113クロマトグラフィーを用いて流体サンプルを分析するための装置であって、前記装置は、 (a)第1流体通路と、 (b)流体のサンプルを分析することができる少なくとも1つの検出器と、 (c)前記第1流体通路を通る流体の流れ特性に実質的に影響を与えることなく、前記第1流体流れ通路から前記検出器へアリコットを輸送するシャトルバルブと、を有する装置。
- 114請求項113に記載の装置であって、前記シャトルバルブを通る前記第1流体通路の少なくとも一部は、実質的に線形または真っ直ぐである、装置。
- 115請求項113に記載の装置であって、前記第1流体通路を通る流体の圧力は、実質的に増加しない、または実質的に一定である、装置。
- 116請求項113に記載の装置であって、前記装置はさらに、前記検出器へ流体のアリコットサンプルを運ぶ第2流体通路を有する、装置。
- 117請求項116に記載の装置であって、前記シャトルバルブを通る前記第2流体通路の少なくとも一部は、実質的に線形または真っ直ぐである、装置。
- 118請求項116に記載の装置であって、前記第2流体通路を通る流体の圧力は、実質的に増加しない、または実質的に一定である、装置。
- 119請求項113に記載の装置であって、前記装置はアリコットディンプルを有し、前記第1流体通がアリコットディンプルに実質的に平行である、装置。
- 120クロマトグラフィーを用いた流体サンプルの分析のための装置であって、前記装置は、 (a)第1流体通路と、 (b)第2流体通路と、 (c)前記サンプルを分析することができる少なくとも1つの検出器と、 (d)シャトルバルブを通る連続な第2流体通路を維持しつつ、前記第1流体通路から前記第2流体通路へアリコットサンプルを輸送するシャトルバルブと、を有する装置。
- 121請求項120に記載の装置であって、前記シャトルバルブを通る前記第1流体通路内のおよび/または前記第2流体通路内の圧力は、実質的に増加せずおよび/または実質的に一定である、装置。
- 122請求項120に記載の装置であって、前記装置はアリコットディンプルを有し、前記第1流体通路は実質的に前記アリコットディンプルに平行である、装置。
- 123クロマトグラフィーを用いて流体サンプルを分析する方法であって、前記方法は、 (a)クロマトグラフィーカラムからの流出物の第1流体を提供するステップと、 (b)サンプルを少なくとも1つの検出器へ運ぶための第2流体を提供するステップと、 (c)シャトルバルブを通る第2流体の連続な流れ通路を維持しつつ、前記第1流体から前記第2流体へアリコットサンプルを取り除くためにシャトルバルブを用いるステップと、 (d)前記アリコットサンプルを観察するために少なくとも1つの検出器を用いるステップと、 (e)検出器応答の変化に応じて、前記第1流体から新しいサンプル画分を画分収集器内に収集するステップと、を有する方法。
- 124請求項123に記載の方法であって、前記第1流体からアリコットサンプルが取り除かれて前記第2流体へ輸送されるときに、前記シャトルバルブを通る前記第1流体の連続な流れ通路が維持される、方法。
- 125請求項123に記載の方法であって、アリコットサンプルが第1流体から取り除かれて前記第2流体へ輸送されるときに、前記第1流体および前記第2流体の連続な流れ通路が維持される、方法。
- 126クロマトグラフィーを用いた流体サンプルを分析する方法であって、前記方法は、 (a)サンプルを含む流れを提供するステップと (b)一般的なキャリア流体を用いて前記流れからサンプルのアリコットを取り除くステップと、 (c)前記一般的なキャリア流体中のアリコとを分析する少なくとも1つの検出器を用いるステップと、 (d)前記一般的なキャリア流体は、有機溶媒および水に混和でき、揮発性でなく、また、非発色性である、ガスまたは液体を有する、方法。
- 127請求項126に記載の方法であって、前記キャリア流体は、イソプロピルアルコール、アセトン、メタノール、エタノール、プロパノール、ブタノール、イソブタノール、テトラヒドロフラン、またはこれらの混合物、を含む、方法。
- 128請求項126に記載の方法であって、前記キャリア流体はイソプロピルアルコールを有する、方法。
- 129フラッシュクロマトグラフィーを用いて流体サンプルを分析するための装置であって、前記装置は、 (a)サンプル内の個別の物質を検出することができる蒸発性粒子検出器と、 (b)検出された物質の検出器応答の変化に応答することができる画分収集器と、を有し、 (c)前記蒸発性粒子検出器は、前記装置中の唯一の検出器である、装置。
- 130請求項129に記載の装置であって、前記蒸発性粒子検出器は、化学組成、化学構造、分子量、またはこれらの組み合わせを検出することができる、装置。
- 131請求項129に記載の装置であって、前記蒸発性粒子検出器は、ELSD、CNLSD、または質量スペクトル計を有する、装置。
- 132フラッシュクロマトグラフィーを用いて流体サンプルを分析する方法であって、前記方法は、 (a)サンプルを観察するための、個別の物質を検出することができる蒸発性粒子検出器を用いるステップと、 (b)物質の検出器応答の変化に応じて、新しいサンプル画分を画分収集器内に収集するステップと、を有し、 (c)前記蒸発性粒子検出器は、サンプルを分析するために用いられる唯一の検出器である、方法。
- 133請求項132に記載の装置であって、前記蒸発性粒子検出器は、化学組成、化学構造、分子量、またはこれらの組み合わせを検出することができる、方法。
- 134請求項132に記載の装置であって、前記蒸発性粒子検出器は、ELSD、CNLSD、または質量スペクトル計を有する、方法。
Independent claims134
106 paragraphs, as filed
The present invention relates to methods and devices for collecting sample fractions and analyzing samples in a chromatography system.
In the art, there is a need for methods for efficiently and effectively analyzing samples and collecting sample fractions in chromatographic systems. Further, in the present technical field, there is a need for an apparatus capable of effectively analyzing a sample and collecting a sample fraction.
<p> The present invention provides a method for efficiently and effectively analyzing a sample and collecting a sample fraction in a chromatography system, and can also effectively analyze a sample and collect a sample fraction. Provide the device.</p>
<p> The present invention relates to a method for analyzing a sample and a method for collecting a sample fraction in a chromatography system. The disclosed methods offer many advantages over known methods of analyzing samples. For example, the disclosed methods of the present invention have at least process variables (eg, flow limiting, total flow rate, temperature, and / or solvent composition) to actively control fluid flow through at least one detector. A splitter pump or shuttle valve can be used so as not to adversely affect the fluid flow through one detector. Also, the disclosed methods of the present invention collect one or more sample fractions in response to one or more detector signals from two or more detectors, as well as a more complete sample given. Two or more detectors can be used to provide the analysis.</p><p> The present invention relates to a method of analyzing a sample and a method of collecting a sample fraction. In one exemplary embodiment, the method of analyzing a sample comprises generating a composite signal from two or more detectors in a liquid chromatography system, the composite signal being a detection response component from each detector. And the method of analyzing a sample has a step of collecting a new sample fraction in a fraction collector in response to changes in the composite signal. In one embodiment, the composite signal comprises (i) a detection response component from at least one optical absorption detector (eg, a UV detector) and (ii) a detection response component from at least one evaporative particle detector. Can have. In one embodiment, a color-developing or non-color-developing solvent can be used as the carrier fluid in the chromatography system. In other embodiments, the composite signal comprises (i) a detection response component comprising two or more detection responses from an optical absorption detector (eg, a UV detector) at two or more specific optical wavelengths, and (ii). It has a detection response component from an evaporative particle detector.</p><p> In a further exemplary embodiment according to the invention, the method of analyzing a sample using chromatography comprises the step of using at least one detector to observe the sample having at least one non-color-developing analysis object. It has a step of collecting a new sample fraction into a fraction collector in response to changes in the detector response of the non-color-developing material. This sample can contain a number of different color-developing or non-color-developing substances. In addition, the mobile phase of transporting the sample can include one or more color-developing or non-color-developing substances.</p><p> In other embodiments, common carrier fluids can be used within the chromatography system, the carrier fluids comprising volatile liquids or various gases. In a further embodiment, the non-destructive detector (eg RI, UV detector, etc.) and the destructive detector (eg, evaporative particle detector, mass spectrometer, spectrophotometer, luminescence spectrometer, NMR, etc.) It can be combined, which makes it possible to detect the identification of various compounds in the sample, for example the presence of chemicals at related peaks.</p><p> In a further embodiment, the method of analyzing the sample involves using at least one detector to observe the sample at two or more optical wavelengths and (i) a change in the detector response at the first wavelength. A new sample fraction is placed in the fraction collector according to (ii) changes in the detector response at the second wavelength and (iii) changes in the composite signal represented by the detector responses at the first and second wavelengths. It has a step to collect and. Changes in a given detector response are, but are not limited to, the threshold of the detector response value, the slope of the detector response with respect to time, the threshold of the slope of the detector response value with respect to time, the slope of the detector response with respect to time. Can include reaching or exceeding a threshold of change in, or any combination thereof. In this embodiment, the method comprises the step of using n sensors in at least one detector to observe n wavelengths over the range of the absorption spectrum, where n is an integer greater than 1. Is. The method also depends on (i) any one change in n detector responses from n sensors, or (ii) a change in the composite signal represented by n detector responses. , Has a step of collecting a new sample fraction in a fraction collector.</p><p> In a further exemplary embodiment, the method of analyzing the sample is (i) a chromatography column, (ii) a T section with a first inlet, a first outlet, and a second outlet, and (iii) a first section of the T section. It has steps to provide a liquid chromatography system with a fraction collector that communicates fluid with the outlet, and (iv) a detector that communicates fluid with the second outlet of the T section, and the method also comprises (v). It has a step of actively controlling the fluid flow through the detector via a splitter pump that communicates the fluid with the second outlet of the T section and the detector. In other exemplary embodiments, a shuttle valve may be used in place of the T section and splitter pump to actively control fluid flow to at least one detector. In one exemplary embodiment, the shuttle valve is a continuous flow shuttle valve capable of removing a very small volume from the sample flow.</p><p> In a further exemplary embodiment of the invention, the method of analyzing a sample fluid using chromatography is with the step of providing a first fluid flowing out of the chromatography column; transporting the sample fluid to at least one detector. To remove a fluid aliquot sample from the first fluid and transport this aliquot to the second fluid, while maintaining a continuous second fluid passage through the shuttle valve. With a step using a shuttle valve; with a step using at least one detector to observe an aliquot sample of the fluid; with a fraction collector a new sample fraction of the first fluid as the detector response changes. Has steps and; to collect within. In one embodiment, a continuous flow path for the first fluid through the shuttle valve is maintained when the aliquot sample of the fluid is removed from the first fluid. In other embodiments, a continuous flow path for both the first and second fluids through the shuttle valve is maintained when the fluid sample aliquot is removed from the first fluid and transported to the second fluid. ..</p><p> In another exemplary embodiment according to the invention, the method of analyzing a fluid sample using chromatography is substantially in the step of providing the first fluid containing the sample; and the flow characteristics of the first fluid through the shuttle valve. With the step of using the shuttle valve to remove the aliquot sample of the fluid from the first fluid without affecting the; with the step of using at least one detector to observe the aliquot sample of the fluid; at least one It has a step of collecting a new sample fraction of the first fluid flow and; in the fraction collector as the detector response changes. The first fluid flow through the shuttle valve is substantially laminar, as the first fluid passage or channel through at least part of the valve is substantially linear or straight. In a further embodiment, the pressure of the first fluid through the shuttle valve is substantially constant and / or does not increase substantially. In other embodiments, the flow rate of the first fluid through the shuttle valve can be substantially constant. In one alternative embodiment, the second fluid is used to carry an aliquot sample of the fluid from the shuttle valve to the detector. The flow of the second fluid through the shuttle valve can be substantially laminar, as the second fluid flow passage or channel through at least part of the valve is substantially linear or straight. In one exemplary embodiment, the pressure of the second fluid through the shuttle valve is substantially constant and / or does not increase substantially. In other embodiments, the flow rate of the second fluid through the shuttle valve can be substantially constant.</p><p> In a further exemplary embodiment, the method of analyzing a sample comprises the steps of providing a liquid chromatography system for a non-destructive system, the liquid chromatography system being (i) a chromatography column, (ii) a system. Two or more non-destructive detectors (eg, optical absorption detectors such as UV detectors), and (iii) two or more non-destructive detectors, in the absence of a destructive detector (eg, a mass spectrometer). It has a destructive detector and a fluid communicative fraction collector, and the method further collects one or more sample fractions in response to detector signals from two or more nondestructive detectors. Have steps.</p><p> In another exemplary embodiment according to the invention, the method of analyzing a sample using flash chromatography is the step of using an evaporative particle detector capable of detecting individual substances to observe the sample. And; with the step of collecting a new sample fraction into the fraction collector in response to changes in the detector response of the substance; where the evaporative particle detector is the only one used to analyze the sample. It is a detector of. Evaporative particle detectors can detect chemical composition, chemical structure, molecular weight, or other chemical or physical properties. This detector can include an ELSD, CNLSD, or mass spectrometer.</p><p> In a further exemplary embodiment, the method of analyzing a sample comprises generating a detector signal from at least one detector in a liquid chromatography system, the detector signal being a function of (i) time. The gradient of the detector response as (ie, the first-order differential of the detector response), (ii) the change in the gradient of the detector response as a function of time (second-order differential of the detector response), (iii) as an optional choice. Occurs in response to reaching or exceeding the threshold of the detector response value, or (iv) any combination of (i) to (iii), preferably including at least (i) or at least (ii). The method further collects one or more sample fractions depending on at least one detector signal from at least one detector.</p><p> In another exemplary embodiment, the method of analyzing a sample comprises collecting the sample fraction in a fraction collector in a liquid chromatography system, the fraction collector (i) at least one. It is configured to recognize, receive, and process one or more signals from one detector, and (ii) collect one or more sample fractions based on one or more signals. To.</p><p> The present invention also relates to an apparatus capable of analyzing a sample. In one exemplary embodiment, the device for analyzing a sample has system hardware configured to generate a composite signal from two or more detectors in a liquid chromatography system, the composite signal. Has a detector response component from each detector, and the device further has a fraction collector configured to collect new sample fractions in response to changes in the composite signal.</p><p> In another exemplary embodiment, the device for analyzing the sample is with at least one detector configured to observe two or more optical wavelengths (eg, UV wavelengths); (i) first wavelength. New samples depending on the change in the detector response in, (ii) the change in the detector response at the second wavelength, or (iii) the change in the composite signal represented by the detector response at the first and second wavelengths. It has a fraction collector configured to collect. As mentioned above, changes in a given detector response include, but are not limited to, changes in the detector response value, reaching or exceeding the threshold of the detector response value, gradient of the detector response value with respect to time, and so on. It can include a threshold for the slope of the detector response value with respect to time, a threshold for the change in the slope of the detector response value with respect to time, or any combination thereof.</p><p> At least one detector can have n sensors for observing at n wavelengths over the range of the absorption spectrum, where n is an integer greater than 1. The fraction collector depends on (i) any one change in the n detector responses from the n sensors, or (ii) a change in the composite response represented by the n detector responses. It is configured to collect new samples. In one embodiment, the device has a single UV detector that has only n sensors or is combined with one or more additional detectors.</p><p> In a further exemplary embodiment, the device that analyzes the sample has system hardware capable of generating a detector signal from at least one detector in a liquid chromatography system, and the detector signal is (1). i) Inclined detector response as a function of time (ie, first-order differential of detector response), (ii) Changes in gradient of detector response as a function of time (ie, second-order differential of detector response), ( iii) Occurs depending on, as an option, reaching or exceeding the threshold of the detector response value, or any combination of (i) to (iii) including (iv) at least (i) or at least (ii). Be made to. The device may further have a fraction collector configured to collect one or more sample fractions in response to a detector signal from at least one detector.</p><p> In another exemplary embodiment according to the invention, an apparatus for analyzing a sample using chromatography can detect at least one detection target that is chromogenic and non-chromogenic in the sample. It has a vessel and a fraction collector that can respond to changes in the detection response of non-color-developing substances. The sample can contain a number of different color-developing and non-color-forming substances. In addition, the mobile phase carrying the sample can include one or more color-developing or non-color-developing substances.</p><p> In a further exemplary embodiment, the apparatus for analyzing the sample is (i) a chromatography column, (ii) a T section with first inlet, first outlet, and second outlet, and (iii) T. Fraction collector that communicates fluid with the 1st outlet of the part, (iv) 1st detector that communicates with the 2nd outlet of the T part and fluid, and (v) 2nd outlet and 1st detector of the T part and fluid It has a splitter pump that is positioned to communicate, and the splitter pump is configured to actively control the fluid flow through the first detector. In other exemplary embodiments, a shuttle valve can be used in place of the T section and splitter pump to actively control fluid flow to at least one detector.</p><p> In a further embodiment of the invention, the apparatus for analyzing a fluid sample using chromatography is a first fluid passage flowing out of a chromatography column or cartridge; at least one detector capable of analyzing the fluid sample. It also includes a shuttle valve capable of transporting an aliquot sample of the fluid from the first fluid passage to the detector without substantially affecting the flow characteristics of the fluid through the first fluid passage. The flow of fluid through the first fluid passage can be substantially laminar, as the first fluid passage or channel is substantially linear or straight where it passes through part of the valve. In a further exemplary embodiment, the pressure of the fluid through the first fluid passage is substantially constant and / or does not increase substantially. In other embodiments, the flow rate of the fluid through the first fluid passage can be substantially constant. In an alternative embodiment, the second fluid passage is used to transport the alicot sample from the shuttle valve to the detector. The fluid flow through the second fluid passage can be substantially laminar, as the second fluid passage or channel is substantially linear or straight where at least part of the valve passes. In an exemplary embodiment, the pressure of the fluid through the second fluid passage is substantially constant and / or does not increase substantially. In a further embodiment, the fluid flow rate is substantially constant through the second fluid passage.</p><p> In a further exemplary embodiment, the apparatus for analyzing the fluid sample using chromatography is with the first fluid passage flowing out of the chromatography column; the fluid sample to at least one detector capable of analyzing the sample. It has a second fluid passage for transporting the fluid; and a shuttle valve for transporting an aliquot sample of fluid from the first fluid passage to the second fluid passage while maintaining a continuous second fluid passage through the shuttle valve. In one embodiment, a continuous first fluid passage through the shuttle valve is maintained when the aliquot sample of fluid is removed from the first fluid passage. In other embodiments, a continuous first or second fluid passage through the shuttle valve is maintained when the fluid aliquot sample is removed from the first fluid passage and transported to the second fluid passage. ..</p><p> In a further embodiment, the apparatus for analyzing the sample is (i) a chromatography column, and (ii) two or more non-destructive detectors without a destructive detector in the system. The apparatus further has (iii) two or more non-destructive detectors and a fluid-communication fraction collector, which is a two or more non-destructive detectors. Collect one or more sample fractions in response to one or more detector signals from the vessel.</p><p> In a further embodiment according to the invention, the apparatus for analyzing a sample using flash chromatography is an evaporative particle detector capable of detecting individual substances in the sample; the detector response of the detected substance. It includes a fraction collector capable of responding to changes, where the evaporative particle detector is the only detector used to analyze the sample. Evaporative particle detectors can detect chemical composition, chemical structure, molecular weight, or other physical or chemical properties. The detector can include an ELSD, CNLSD, or mass spectrometer.</p><p> In another exemplary embodiment, the device for analyzing the sample has a fraction collector in a liquid chromatography system, which fraction collector is (i) one from at least one detector. It is configured to recognize, receive, and process one or more signals, and (ii) collect one or more sample fractions based on this one or more signals.</p><p> The methods and devices of the present invention have at least one detector. Suitable detectors are non-destructive detectors such as, but not limited to, UV detectors, RI detectors, conductivity detectors, fluorescence detectors, light scattering detectors, viscous detectors, and polarization detectors. (Ie, detectors that do not consume or destroy the sample during detection); and / or evaporative particle detectors such as, for example, Evaporative Light Scattering Detector (ELSD) and Aggregate Nucleating Light Scattering Detector (CNLSD). EPDs), corona discharge detectors, mass spectrometers, destructive detectors such as atomic absorptiometers (ie, detectors that consume or destroy samples when detecting) can be included. For example, the apparatus of the present invention includes at least one UV detector, at least one evaporative light scattering detector (ELSD), at least one mass spectrometer, at least one aggregated nucleating light scattering detector (CNLSD), at least. It can include one corona discharge detector, at least one refractometer detector (RID), at least one fluorescence detector (FD), a chiral detector (CD), or a combination thereof. In one exemplary embodiment, the detector has one or more evaporative particle detectors (EPDs), which allow the use of chromogenic or non-chromogenic solvents as the mobile phase. In a further embodiment, a non-destructive detector can be combined with a destructive detector, which is the property, molecular weight, chemical structure, elemental composition of various materials, such as the presence of species associated with peaks. , Allows detection of sample chiral symmetry.</p><p> The present invention further relates to a computer-readable medium containing computer-executable instructions for performing one or more of the steps of any exemplary method described herein. This computer-readable medium can be used to load application code onto a device or device component, such as a device component as described herein, (i) an interface to the operator. Provide and / or (ii) provide logic to perform one or more of the steps of the methods described herein.</p><p> These and other features and advantages of the present invention will become apparent by reference to the detailed description of the embodiments disclosed below and the appended claims.</p>
<figref num="1">It is a figure which shows the exemplary liquid chromatography system of this invention which has a splitter pump for actively controlling a fluid flowing to a detector.</figref><figref num="2">It is a figure which shows another example of the liquid chromatography system of this invention which has a splitter pump and a detector.</figref><figref num="3A">It is a figure which shows an example of the liquid chromatography system of this invention which has a shuttle valve and a detector.</figref><figref num="3B">It is a figure which shows the operation of the exemplary shuttle valve which can be suitably used for this invention.</figref><figref num="3C">It is a figure which shows the operation of the exemplary shuttle valve which can be suitably used for this invention.</figref><figref num="4">It is a figure which shows the exemplary liquid chromatography system of this invention which has a splitter pump and two detectors.</figref><figref num="5">It is a figure which shows the exemplary liquid chromatography system of this invention which has two splitter pumps and two detectors.</figref><figref num="6">It is a figure which shows the exemplary liquid chromatography system of this invention which has a shuttle valve and two detectors.</figref><figref num="7">It is a figure which shows the exemplary liquid chromatography system of this invention which has two shuttle valves and two detectors.</figref><figref num="8">It is a figure which shows the exemplary liquid chromatography system of this invention which has a splitter pump, an evaporative light scattering detector (ELSD), and an ultraviolet (UV) detector.</figref><figref num="9">It is a figure which shows the other exemplary liquid chromatography system of this invention which has a splitter pump, ELSD and a UV detector.</figref><figref num="10A">It is a figure which shows the operation of the shuttle valve which can be suitably used for this invention.</figref><figref num="10B">It is a figure which shows the operation of the shuttle valve which can be suitably used for this invention.</figref><figref num="10C">It is a figure which shows the operation of the shuttle valve which can be suitably used for this invention.</figref><figref num="11">It is a figure which shows the chromatogram generated from the separation of two composite mixtures using the exemplary chromatography system of this invention.</figref>
Specific embodiments of the invention are described to facilitate understanding of the principles of the invention, and specific language is used to describe the embodiments. However, it should be understood that the use of particular language is not intended to limit the scope of the invention. It is expected that variants, further modifications, and further applications of the principles of the invention discussed will be common to those skilled in the art of the invention.
As used herein and in the appended claims, the singular representation includes multiple references unless the context makes it clear. Thus, for example, the term "solvent" is intended to include multiple solvents, including one or more solvents known to those of skill in the art and their equivalents.
For example, it is used in the description of embodiments of the present disclosure that modify the amount, concentration, volume, process temperature, process time, recovery or yield rate, flow rate, and similar values of the components of the complex, as well as the range. The term "about" refers to, for example, numerical variability in quantities that can occur through typical measurement and processing procedures, variability due to errors in these procedures, variability due to differences in the components used to perform the method. , And the variation by approximation. The term "about" also includes the amount of change in formulation over time at a particular initial concentration and mixture, and the amount of change due to mixing or processing the formulation at a particular initial concentration or mixture. Whether or not modified by the word "about", the appended claims include an equal range of these amounts.
As used herein, the term "chromatography" means a physical separation method in which the components to be separated are distributed between two phases, one of which is static (static phase). Yes, the other moves in one direction (movable phase).
As used herein, the term "liquid chromatography" refers to the separation of a mixture by passing the dissolved liquid mixture through a column having a quiescent phase into the "moving phase", the analyte (ie, the target substance). Is separated from other molecules in the mixture so that it is sequestered.
As used herein, the term "moving phase" means a sample to be separated and / or analyzed, and a liquid fluid, gas, or supercritical fluid containing a solvent, where the solvent has an analyte that passes through a column. Move the sample. The mobile phase travels through a chromatography column or cartridge (ie, the container that houses the rest phase), where the analyte in the sample interacts with the rest phase and is separated from the sample.
As used herein, the term "quiescent phase" means a material immobilized on a column or cartridge, which selectively adsorbs an analyte from a sample within the mobile phase and through a column having a quiescent phase. The mixture is separated by passing the dissolved fluid mixture through the "moving phase" so that the analytes measured from other molecules in the mixture are separated and sequestered.
As used herein, the term "flash chromatography" means separation by passing a mixed fluid dissolved in a "moving phase" under pressure through a column having a stationary phase, which is within the mixture. Separate the analyte (ie, the target substance) from other molecules so that it can be sequestered.
As used herein, the term "shuttle valve" means a control valve that controls the supply of fluid from one or more sources to other locations. The shuttle valve can utilize rotary or linear motion to move the sample from the fluid to the other.
As used herein, the term "fluid" means a gas, liquid, or supercritical fluid.
As used herein, the term "laminar flow" means the smooth, regular motion of a fluid, in which there is no turbulence in the laminar flow, and any given side flow is a side of another neighborhood. It moves almost parallel to the flow.
As used herein, the term "substantial" means within a reasonable amount, about 0% to about 50%, about 0% to about 40%, about 0% of the absolute value. Includes amounts that vary from about 30%, from about 0% to about 20%, or from about 0% to about 10%.
The present invention relates to a method of analyzing a sample and collecting a sample fraction. The present invention further relates to an apparatus capable of analyzing a sample and collecting a sample fraction. The present invention further relates to computer software for use within a device or device component capable of analyzing a sample and collecting a sample fraction, wherein the device is one or more described herein. Be able to perform the steps of the method.
Below is a description of an exemplary method for analyzing a sample and an exemplary device capable of analyzing the sample.
1. How to analyze the sample The present invention relates to a method of analyzing a sample and collecting a sample fraction. The method of analyzing this sample can have a number of process steps, some of which are described below.
A. Active control of fluid flow to the detector In some embodiments of the invention, the method of analyzing a sample comprises the step of dynamically controlling the fluid flow to the detector via a splitter pump or shuttle valve. An exemplary liquid chromatography system demonstrating such method steps is illustrated in FIG. As shown in FIG. 1, an exemplary liquid chromatography system 10 has (i) a chromatography column 11 and (ii) a T section 12 comprising a first inlet 21, a first outlet 22, and a second outlet 23. (Iii) Fraction collector 14 that communicates fluid with the first outlet of T section 12, (iv) First detector 13 that communicates fluid with the second outlet 23 of T section 12, and (v) T section. It has a splitter pump 15 positioned to communicate fluid with a second outlet 23 of twelve and a first detector 13.
In this exemplary system, the splitter pump 15 actively controls the fluid flow to the first detector 13. As used herein, the expression "actively controlled" is given in a given splitter pump or shuttle valve, even if there is a change in flow rate elsewhere in the liquid chromatography system. It is the ability to control the flow of fluid through a detector. Unlike "passive" flow splitters, which simply divide the fluid flow, the splitter pumps and shuttle valves used in the present invention have, for example, flow limiting, total flow rate, temperature, and / or solvent composition. Control the fluid flow to at least one detector, regardless of possible fluctuations in the fluid flow in the liquid chromatography system.
The steps of actively controlling the fluid flow to a given detector include, for example, the step of sending a drive signal to the splitter pump or shuttle valve, (i) driving the splitter pump or shuttle valve, and (ii) the splitter. Stop driving the pump or shuttle valve, (iii) change one or more flows, and / or change the pressure setting of the splitter pump or shuttle valve, or (iv) (i) through (iii). Make any combination. Appropriate flow and pressure settings are, but are not limited to, (i) valve position, (ii) splitter pump or shuttle valve pressure, (iii) air pressure on the valve, or (iv) (i) to ( iii) Includes any combination up to. Typically, the drive signal is in the form of, for example, an electrical signal, a gas signal, a digital signal, or a wireless signal.
As shown in FIG. 1, in an exemplary liquid chromatography system 10, the step of actively controlling the fluid flow to the detector 13 is to pump the fluid from the T section 12 to the detector 13. It has a step using a splitter pump 15. In other embodiments, the step of actively controlling the fluid to the detector may include the step of using a splitter pump to draw the fluid through the detector.
FIG. 2 shows an exemplary liquid chromatography system 20, which includes a chromatography column 11; a T section 12 having a first inlet 21, a first outlet 22, and a second outlet 23; Fraction collector 14 that communicates fluid to the first outlet 22 of 12; first detector 13 that communicates fluid to the second outlet 23 of the T section 12; fluid from the second outlet 23 of the T section 12 through the detector 13. It has a splitter pump 15, which is positioned to pull out.
In some desirable embodiments, a shuttle valve, such as the exemplary shuttle valve 151 shown in FIG. 3A-3C, is used to actively control fluid flow to a detector, such as detector 131. As shown in FIG. 3A, an exemplary liquid chromatography system 30 comprises a chromatography column 11; a chromatography cartridge inlet 111, a fraction collector outlet 114, a gas or liquid inlet 115, and a detector outlet 113. Shuttle valve 151 and; Fraction collector 14 that communicates fluid to fraction collector outlet 114 of shuttle valve 151; First detector 131 that fluid communicates to detector outlet 113 of shuttle valve 151; Gas of shuttle valve 151 Alternatively, it has a fluid supply unit 152, which supplies a fluid to the liquid inlet 115.
In a further embodiment of the invention, the method of analyzing a sample using chromatography is a step of providing a first fluid flowing out of a chromatography column; a second for transporting the fluid sample to at least one detector. With the steps to provide the fluid; remove the sample aliquot from the first fluid, transport this aliquot to the second fluid, and use the shuttle valve to maintain a continuous path through the shuttle valve of the second fluid. Steps to use; Steps to use at least one detector to observe an aliquot of the fluid sample; Collect a new sample fraction of the first fluid in the fraction collector as the detector response changes. Has steps to do. In one embodiment, the continuous flow path through the shuttle valve of the first fluid is maintained when the aliquot of the fluid sample is removed from the first fluid. In another embodiment, a continuous flow path through both the first and second fluid shuttle valves is maintained when the aliquot of the fluid sample is removed from the first fluid and transported to the second fluid.
In another exemplary embodiment of the invention, the method of analyzing a fluid sample using chromatography is substantially the step of providing a first fluid containing the sample; substantially the flow characteristics of the first fluid through the shuttle valve. A step using a shuttle valve to remove the aliquots of the fluid sample from the first fluid, which would affect; a step using at least one detector to observe the aliquots of the fluid sample; at least one detector It has a step of collecting a new sample fraction of the first stream into a fraction collector, depending on the change in the response of. The flow of the first fluid through the shuttle valve can be substantially laminar by allowing the first fluid passage or channel to be substantially linear or straight through at least part of the valve. In a further embodiment, the pressure of the first fluid through the shuttle valve remains substantially constant and / or the pressure does not increase substantially. In other embodiments, the flow rate of the first fluid can be made substantially constant through the shuttle valve. In an alternative embodiment, a second fluid is used to transport the aliquot of the liquid sample from the shuttle valve to the detector (s). The flow of the second fluid through the shuttle valve can be substantially laminar because the second fluid passage or channel is substantially linear or straight through at least part of the valve. In an exemplary embodiment, the pressure of the second fluid through the shuttle valve is substantially constant and / or the pressure does not increase substantially. In other embodiments, the flow rate of the second fluid can be substantially constant through the shuttle valve.
3B and 3C show the operation of the shuttle valve in one exemplary embodiment within a given liquid chromatography system. As shown in FIG. 3B, the shuttle valve 151 is provided with a chromatography cartridge inlet 111 that provides fluid flow from the chromatography column (eg, column 11) to the shuttle valve 151; with an inflow sample alicot volume 116; from the shuttle valve 151. With the fraction collector outlet 114, which provides fluid flow to the fraction collector (eg, fraction collector 14); provides gas (eg, air, nitrogen, etc.) or liquid (eg, alcohol) flow through a portion of the shuttle valve 151. It has a gas or liquid inlet 115; an outflow sample aliquot volume 117; and a detector outlet 113 that provides fluid flow from a shuttle valve 151 to a detector (eg, a detector 131 such as an ELSD).
When a fluid flows from the chromatographic cartridge inlet 111 to the fraction collector outlet 114 through the shuttle valve 151, the inflow sample aliquot volume 116 is filled with a specific volume of fluid, which is referred to herein as the sample aliquot 118. (Shown as a shaded area in Figure 3B). At the desired time, the shuttle valve 151 transports the sample aliquot 118 within the inflow sample alicot volume 116 into the outflow sample alicot volume 117 shown in FIG. 3C. When the sample aliquot 118 is transported into the spill sample alicot volume 117, gas or liquid from the inlet 115 passing through the spill sample alicot volume 117 will detect the sample alicot 118 through the detector outlet 113. Transport to 131 (eg ELSD).
Shuttle valve 151 can be programmed to remove sample aliquots (eg, sample aliquot 118) from a sample at a desired sampling frequency for transport to at least one detector. In one exemplary embodiment, the sampling frequency is at least one sample aliquot every 10 seconds (or at least one sample alicot every 5 seconds, or at least one sample alicot every 3 seconds, or It can be at least one sample aliquot every 2 seconds, or at least one sample alicot every 0.5 seconds, or at least one sample alicot every 0.1 seconds).
FIG. 10A-10C shows an exemplary shuttle valve of the present invention and also shows the operation of the shuttle valve within a given chromatography system. As shown in FIG. 10A, the shuttle valve 151 has a chromatography cartridge inlet 111 that provides fluid flow from the chromatography column (eg, column 11) to the shuttle valve 151; and a channel 117 that connects the inlet 111 to the outlet 114. With the inflow sample aliquot volume 118 in the dimple 116 of the dynamic body 119; with the fraction collector outlet 114 that provides the fluid flow from the shuttle valve 151 to the fraction collector (eg fraction collector 14); With a gas or liquid inlet 115 that provides a flow of gas (eg air, nitrogen, etc.) or liquid (eg alcohol) through the shuttle valve 151; with an outflow sample alicot volume 118 in the dimple 116; connecting the inlet 115 to the outlet 113. It has a channel 120 and a detector outlet 113 that provides fluid flow from the shuttle valve 151 to the detector (eg, a detector 131 such as an ELSD).
As the fluid flows through the shuttle valve 151 from the chromatography cartridge to the inlet 111 and through the channel 117 to the fraction collector outlet 114, the inflow sample alicot volume 118 in the dimple 116 becomes with the sample alicot 118. Filled with the volume of the particular fluid mentioned (shown in the shaded area of Figure 10A). At the desired time, the shuttle valve 151 transports the sample aliquot 118 in the dimples 116 removed from the channel 117 to the channel 120 by rotating the dimples 116 in the dynamic body 119 through the dimple rotation passage 121. To do. When the sample aliquot 118 is transported to the channel 120, the gas or liquid flowing from the inlet 115 through the channel 120 transports the sample alicot 118 to the detector 131 (eg ELSD) via the detector outlet 113. Another advantage of the shuttle valve according to the invention relates to the hydrodynamic design of the channel through the valve. The flow through channels 117 and 120 is continuous to minimize back pressure in the chromatography system. This is to place channels 117 and 120 within the static body 122 so that the flow through the shuttle valve 151 is continuous (shown in FIG. 10B) no matter where the dynamic body 119 is located. Achieved by. As shown in FIG. 10A, at least part of the sample flow channel 117 and the detector flow channel 120 is substantially in plane or circumference, which reduces turbulence and further increases pressure through the valve. To minimize. In addition, at least part of the sample flow channel 117 and the detector flow channel 120 are substantially parallel when adjacent to the dimple 116, which further limits turbulence and limits pressure buildup in the valve. .. This includes a configuration that does not increase the pressure in the valve, 50 psi or more, preferably 30 psi or more, more preferably 20 psi or more, still more preferably 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 psi. After that Above, it includes a configuration that does not increase the pressure. The dimples 116 are located within the dynamic body 119 and communicate fluidly with the surface of the dynamic body, which is adjacent to the static body 122. Thus, when the dynamic body 119 is in the first position, the dimples 116 communicate with the sample flow channel 117, and when the dynamic body 119 moves to the second position, the dimples 116 fluid into the detector flow channel 120. Communicate. The dimples 116 can have any shape, but are shown as concave hemispheres. Further, the dimple 116 can have any size. In one exemplary embodiment, the dimples can be extremely small in size (eg, less than 2000nL, preferably less than about 500nL, more preferably less than about 100nL, more preferably less than about 1nL), but from 1nL. It can contain any dimensions up to 2000nL, which allows for rapid sampling. In addition, the dimensions of the small dimples 116 allow for a very short dimple rotation passage 121, which significantly reduces the surface friction between the dynamic body 119 and the static body 122 and requires maintenance of the shuttle valve 151. Longer service life (for example, 10 million times before maintenance). A rotary motion shuttle valve is shown in Figure 10A-10C, but a linear motion shuttle valve, or an equivalent shuttle valve, can also be used in the present invention. It can be extremely small (eg, less than 2000nL, preferably less than about 500nL, more preferably less than about 100nL, more preferably less than about 1nL), but can include any size from 1nL to 2000nL. , This allows for quick sampling. In addition, the dimensions of the small dimples 116 allow for a very short dimple rotation passage 121, which significantly reduces the surface friction between the dynamic body 119 and the static body 122 and requires maintenance of the shuttle valve 151. Longer service life (for example, 10 million times possible before maintenance). A rotary motion shuttle valve is shown in FIGS. 10A-10C, but a linear motion shuttle valve, or an equivalent shuttle valve, can also be used in the present invention. It can be extremely small (eg, less than 2000nL, preferably less than about 500nL, more preferably less than about 100nL, more preferably less than about 1nL), but can include any size from 1nL to 2000nL. , This allows for quick sampling. In addition, the dimensions of the small dimples 116 allow for a very short dimple rotation passage 121, which significantly reduces the surface friction between the dynamic body 119 and the static body 122 and requires maintenance of the shuttle valve 151. Longer service life (for example, 10 million times before maintenance). A rotary motion shuttle valve is shown in FIGS. 10A-10C, but a linear motion shuttle valve, or an equivalent shuttle valve, can also be used in the present invention.
The shuttle valve 151 can be programmed to remove a sample aliquot (eg, a sample aliquot 118) from a sample at a desired sampling frequency for transport to at least one detector. In one exemplary embodiment, the sampling frequency is at least 1 sample alicot every 10 seconds (or at least 1 sample alicot every 5 seconds, or at least 1 sample alicot every 3 seconds, or at least every 2 seconds. It can be 1 sample alicot, or 1 sample alicot every 0.5 seconds, or at least 1 sample alicot every 0.1 seconds). This shuttle valve is further described in the pending US Provisional Patent Application ____, which is incorporated herein by reference in its entirety.
In other embodiments, common carrier fluids, including volatile liquids and various gases, can be used in the chromatography system to carry the sample to the detector. As shown in FIG. 3A, the carrier fluid from the fluid supply 152 enters the shuttle valve 151 at the inlet 115, where the carrier sphere takes out the sample aliquot 118 and through the outlet 113 the detector 131. Carry to. The sample aliquot should not settle in the carrier fluid of the valve, and the associated piping should not be blocked so that the sample covers the wall of the flow path so that some or all of the sample does not reach the detector. Not. The composition of samples in flash chromatography is very diverse and includes a variety of chemicals including inorganic molecules, organic molecules, polymers, peptides, proteins, and oligonucleotides. Solubility in various solvents depends on the class of compound. Also, compatibility with the detector limits the types of carrier fluids that can be used. For example, in the case of UV detectors, the solvent should be non-coloring at the detection wavelength. For evaporated particle detection (EPD) techniques (ELSD, CNLSD, mass spectrum, etc.), the solvent should be able to easily evaporate at a temperature well below the melting point of the sample. In addition, the carrier fluid should be miscible with the sample flowing between the valve inlet 111 and the fraction collector outlet 114. For example, if hexane is used in one flow path, water should not be used in another. This is because they are not miscible. The carrier fluid proposed above should be customized when changing the separation solvent. This is time consuming and impractical. According to an exemplary embodiment of the invention, this problem can be avoided by using a volatile, non-coloring solvent that is miscible with an organic solvent and water. A volatile, non-coloring, intermediately polar solvent such as isopropyl alcohol (IPA) can be used as the carrier fluid. IPA is almost all It is miscible with all solvents, is non-coloring at common UV detection wavelengths, and easily evaporates at low temperatures. In addition, IPA dissolves a wide range of chemicals and species. Therefore, IPA is suitable as a carrier fluid for virtually any sample type. Other carrier fluids can include acetone, methanol, ethanol, propanol, butanol, isobutanol, tetrahydrofuran and the like. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. It is non-color-developing at typical UV detection wavelengths and easily evaporates at low temperatures. In addition, IPA dissolves a wide range of chemicals and species. Therefore, IPA is suitable as a carrier fluid for virtually any sample type. Other carrier fluids can include acetone, methanol, ethanol, propanol, butanol, isobutanol, tetrahydrofuran and the like. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. It is non-color-developing at typical UV detection wavelengths and easily evaporates at low temperatures. In addition, IPA dissolves a wide range of chemicals and species. Therefore, IPA is suitable as a carrier fluid for virtually any sample type. Other carrier fluids can include acetone, methanol, ethanol, propanol, butanol, isobutanol, tetrahydrofuran and the like. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. Evaporates easily. In addition, IPA dissolves a wide range of chemicals and species. Therefore, IPA is suitable as a carrier fluid for virtually any sample type. Other carrier fluids can include acetone, methanol, ethanol, propanol, butanol, isobutanol, tetrahydrofuran and the like. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. Evaporates easily. In addition, IPA dissolves a wide range of chemicals and species. Therefore, IPA is suitable as a carrier fluid for virtually any sample type. Other carrier fluids can include acetone, methanol, ethanol, propanol, butanol, isobutanol, tetrahydrofuran and the like. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. PA is suitable as a carrier fluid for virtually any sample type. Other carrier fluids can include acetone, methanol, ethanol, propanol, butanol, isobutanol, tetrahydrofuran and the like. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. PA is suitable as a carrier fluid for virtually any sample type. Other carrier fluids can include acetone, methanol, ethanol, propanol, butanol, isobutanol, tetrahydrofuran and the like. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. Lochlann and the like can be included. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. Lochlann and the like can be included. In an exemplary alternative embodiment, gas can be used as the carrier fluid. Precipitation does not occur because the sample remains in the separation solvent or in the mobile phase through the shuttle valve and subsequent detectors. Similarly, the separation solvent or mobile phase does not mix with other solvents so that miscibility issues do not arise. Since the carrier is a gas, volatility is not an issue. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. So volatility doesn't matter. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. So volatility doesn't matter. In addition, many gases are non-coloring and are suitable for UV detection. When the gas is used as a carrier, the sample aliquot 118 is fed from the valve 151 to the detector 131 as a separate slag sandwiched between the gas pockets 123, as shown in FIG. 10C. The use of gas as the carrier fluid also has other advantages. For example, when used with an evaporative light scattering detector, or other detection technique, the sample is atomized here and the gas can be used to transport the sample and also to atomize the sample. It can also eliminate the need for a separate fog-forming gas supply. Moreover, since the gas does not need to evaporate, the ambient drift tube temperature can be used to eliminate the need for a drift tube heater. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. To. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used. To. Since the sample that evaporates at high temperature is in a solid or liquid state as it passes through the drift tube, a wide range of samples can be detected. Various gases can be used as carrier fluids, which can include air, nitrogen, helium, hydrogen, and carbon dioxide. Supercritical fluids such as supercritical carbon dioxide can also be used.
B. Detection of sample components in fluid flow The method of the present invention can further use at least one detector to detect one or more sample components in the fluid stream. Suitable detectors for use in the liquid chromatography system of the present invention include, but are not limited to, non-destructive and / or destructive detectors. Suitable detectors include, but are not limited to, non-destructive detectors (ie, detectors that do not consume or destroy the sample during detection), such as UV detectors, RI detectors, conductive detectors. , Fluorescence detectors, light scattering detectors, viscometers, polarization meters, etc., and suitable detectors include destructive detectors (ie, detectors that consume or destroy samples during detection), eg , Evaporative Particle Detectors (EPDs), Corona Discharge Detectors, Mass Spectrum Detectors, Atomic Absorption Detectors, etc., such as Evaporative Light Scattering Detectors (ELSD) and Aggregated Nucleating Light Scattering Detectors (CNLSD). .. For example, the apparatus of the present invention has at least one UV detector, at least one evaporative light scattering detector (ELSD), at least one mass spectrum detector (MS), and at least one aggregated nucleating light scattering detector (CSLSD). ), At least one corona discharge detector, at least one differential refractometer (RID), at least one fluorescence detector (FD), at least one chiral detector (CD), or a combination thereof. .. In an exemplary embodiment, the detector can include one or more Evaporative Particle Detectors (EPDs), which allows the use of luminescent and non-luminescent solvents as fluidized beds. In a further embodiment, the non-destructive detector can be combined with a destructive detector, which is a sample, such as chemical properties, chemical structure, molecular weight, etc., associated with each chromatographic peak. The properties of various compounds can be detected. When combined with mass spectrum detection, the chemical structure and / or molecular weight of the fraction can be detected at the time of detection, and the desired fraction is continuously identified. In the current system, the chemical identity and structure of the fractions are treated
Regardless of the type of detector used, a given detector provides one or more detection responses, which generate a signal and one or more in the liquid chromatography system described herein. Used to transmit to components (eg fraction collectors, other detectors, splitter pumps, shuttle valves, T-shaped parts, etc.). Typically, a change in the response of a given detector triggers signal generation and transmission. In the present invention, the change in the response value of a given detector, which triggers the generation of a signal and transmitted to one or more components, is not limited, but the change in the response value of the detector and the threshold value of the detection value. Includes reaching or exceeding, the time gradient of the detected response value, the threshold of the time gradient of the detected response value, the change of the time gradient of the detected response value, the change of the threshold of the time gradient of the detected response value, or a combination thereof.
In some exemplary embodiments, the liquid chromatography system of the present invention has at least two detectors as shown in FIG. An exemplary liquid chromatography system 40 shown in FIG. 4 has a chromatography column 11; a first inlet 21, a first outlet 22, and a second outlet 23 in a T section 12; a first outlet 22 of the T section 12. Fraction collector 14 that communicates fluid; 1st detector 13 that communicates fluid to the 2nd outlet 23 of T section 12; Actively controls the fluid flow from the 2nd outlet 23 of T section 12 to the 1st detector 13. It has a splitter pump 15; and a second detector 16 that communicates fluid with the second outlet 23 of the T section.
In the presence of more than one detector, the liquid chromatography system provides the operator with many analytical options. For example, in the exemplary liquid chromatography system 4 shown in FIG. 4, the method of analyzing a sample is optical, such as first detector 13 (eg ELSD) and / or second detector 16 (eg UV detector). The absorption detector) transmits one or more signals to the fraction collector 14, causing the fraction collector 14 to collect a new sample fraction. One or more signals from the first detector 13 and / or the second detector 16 is a single signal from the first detector 13 or the second detector 16, the first detector 13 and the second detector. It can include two or more signals from 16 or a composite signal from the first detector 13 and the second detector 16. In the exemplary liquid chromatography system 40 shown in FIG. 4, the method of analyzing a sample further transmits a signal from the second detector 16 to the splitter pump 15 to detect the sample component in the fluid flow. 2 The response of the detector 16 can have a step of starting or stopping the fluid flow to the first detector 13.
In another exemplary embodiment, the liquid chromatography system of the present invention has at least two detectors and at least two splitter pumps, as shown in FIG. An exemplary liquid chromatography system 50 shown in FIG. 5 includes a chromatography column 11; a first T section 12 with a first inlet 21, a first outlet 22, and a second outlet 23; a second outlet of the first T section 12. 1st detector 13 that communicates fluid with 23; 1st splitter pump 15 that actively controls the fluid flow from the 2nd outlet 23 of the 1st T part 12 to the 1st detector 13; 1st inlet 31, 1st outlet 32, and a second T section 18 with a second outlet 33; a second detector 16 that communicates fluid to the second outlet 33 of the second T section 18; from the second outlet 33 of the second T section 18 to the second detector 16. It has a second splitter pump 17 that actively controls the fluid flow; a fraction collector 14 that communicates the fluid with the first outlet 32 of the second T section 18.
As mentioned above, the liquid chromatography system of the present invention has one or more shuttles in place, as shown in FIGS. 6 and 7, in order to actively control the fluid flow to at least one detector. It can be equipped with a valve or a combination of one or more T-part / splitter pumps. As shown in FIG. 6, an exemplary liquid chromatography system 60 is a shuttle with a chromatography column 11; a chromatography cartridge inlet 111, a fraction collector outlet 114, a gas or liquid inlet 115, and a detector outlet 113. Valve 151; Fraction collector of shuttle valve 151 Fluid collector 14; Fluid communication to detector outlet 113 of shuttle valve 151 First detector 131; Gas or liquid inlet 115 of shuttle valve 151 It has a fluid supply unit 152; a second detector 161; which communicates the fluid with the detector outlet 113 of the shuttle valve 151.
As shown in FIG. 7, an exemplary liquid chromatography system 70 comprises a chromatography column 11; a chromatography cartridge inlet 111, a fraction collector outlet 114, a gas or liquid inlet 115, and a detector outlet 113. 1 Shuttle valve 151; First detector 131 that communicates fluid to the detector outlet 113 of the shuttle valve 151; Fluid supply unit 152 that supplies fluid to the gas or liquid inlet of the shuttle valve 151; Chromatography cartridge inlet 121, Fraction collection Second shuttle valve 171 with instrument outlet 124, gas or liquid inlet 125, and detector outlet 123; second detector 161 with fluid communication to detector outlet 123 of shuttle valve 171; gas or liquid inlet 125 of shuttle valve 171 It has a fluid supply unit 172; and a fraction collector 14; which communicates the fluid with the fraction collector outlet 124 of the shuttle valve 171.
In these exemplary embodiments, ie, the exemplary liquid chromatography systems 50 and 70, the method of analyzing a sample is further via a second splitter pump 17 (or second shuttle valve 171) to a second detector 16. It has steps to actively control fluid flow to (or second detector 161), as well as first detector 13 (or first shuttle valve 151) via first splitter pump 15 (or first shuttle valve 151). 1 Has a step to actively control the fluid flow to the detector 131). Although not shown in FIG. 5, the first splitter pump 15 and / or the second splitter pump 17 has the first detector 13 and the second detector 16 in an exemplary liquid chromatography system 50, respectively. It should be understood that it can be positioned to push or pull the fluid through.
In some exemplary embodiments, one or more optical absorption detectors, such as one or more UV detectors, are used to observe the detector response and response changes at one or more wavelengths of the absorption spectrum. Can be used. In these embodiments, one or more light sources can be used in combination with a single detector or multiple sensors in multiple detectors to detect light absorption by the sample at multiple wavelengths. For example, one or more UV detectors can be used to observe the detector response and response change at one or more wavelengths over the entire UV absorption spectrum.
In one exemplary method of analyzing a sample, the method comprises the step of using an optical absorption detector, such as a UV detector, which has n pieces across all wavelengths of the UV absorption spectrum. It has n sensors for observing samples at wavelengths, and this method also has (i) any one change in n detection responses at n UV wavelengths, or (ii) n detections. It has a step of collecting a new sample fraction according to the change in the composite response represented by the response. If there are n sensors or multiple detectors, they are relative to each other so as to affect signal timing for fraction collectors and / or other system elements (eg other UV detectors). It can be positioned as desired.
When using full-spectrum UV (or other spectral range) analysis, the spectrum can be divided into the desired number of regions of interest (eg 200 nm to 400 nm divided by 5 nm). Significant temporal changes in each spectral range can be monitored. A sharp drop in the received light energy within a given range (eg, a drop in the first and second derivatives of the detection response) indicates the arrival of a substance that absorbs light at a wavelength in the given range of interest. In this exemplary embodiment, each range can be made smaller to increase accuracy, and alternative, each range can be made larger to reduce the computational burden (ie, per hour). Less computation and less memory required).
In other exemplary embodiments, a plurality of different types of detectors can be used to observe different detection responses and to observe changes in detection responses within a given system. In the liquid chromatography system 80 shown in the exemplary FIG. 8, an evaporative particle detector (EPD) such as an evaporative light scattering detector (ELSD) (ie, first detector 13) alone or UV detection. Used in combination with a device (ie, second detector 16). An exemplary liquid chromatography system 80 further comprises a chromatography column 11; with a T section 12 having a first inlet 21, a first outlet 22, and a second outlet 23; with a fraction collector 14; with a T section 12. An EPD 13 that communicates fluid to the second outlet; a splitter pump 15 that actively controls the fluid flow to the EPD 13; and a UV detector that communicates fluid to the first outlet 22 of the T section 12; Have. In this exemplary embodiment, the use of an evaporative particle detector offers several benefits. The non-color-developing movable phase must be used with UV detection, otherwise background absorption of the movable phase hides the sample signal. This eliminates the use of solvents with significant color development such as toluene, pyridine and others. For evaporative particle detection, the color development of the mobile phase is not important. Color-developing solvents can be used to detect evaporative particles as long as the mobile phase is more volatile than the sample. This opens up the opportunity to improve separation by using a highly selective chromogenic solvent as the mobile phase. In addition, the UV detector will not detect non-color-developing sample components. Fractions collected based solely on UV detection may contain one or more undefinable non-coloring components, which compromises the purity of the fraction. Conversely, non-color-developing samples are completely overlooked by UV detectors and sent directly to disposal or collected within what is supposed to be sample-free (blank fractions). Net results result in reduced productivity, contaminated fractions, or loss of valuable sample components. When EPD (eg ELSD) is used alone or with UV detection in a flash system, chromogenic and non-chromogenic components are detected and collected to improve fraction purity. A flash system that uses the UV system alone overlooks sample components or inaccurately determines the pure fraction, many flash users check the purity, and the blank fraction is really blank. The collected fractions will be screened by thin film chromatography to see if. This is a time-consuming separation procedure that slows down the work. These fractions found to contain one or more components often require a second chromatography step to properly separate the components. Alternatively, when used with UV detection, chromogenic and non-chromogenic components are detected and collected to improve the purity of the fraction. A flash system that uses the UV system alone overlooks sample components or inaccurately determines the pure fraction, many flash users check the purity, and the blank fraction is really blank. The collected fractions will be screened by thin film chromatography to see if. This is a time-consuming separation procedure that slows down the work. These fractions found to contain one or more components often require a second chromatography step to properly separate the components. Alternatively, when used with UV detection, chromogenic and non-chromogenic components are detected and collected to improve the purity of the fraction. A flash system that uses the UV system alone overlooks sample components or inaccurately determines the pure fraction, many flash users check the purity, and the blank fraction is really blank. The collected fractions will be screened by thin film chromatography to see if. This is a time-consuming separation procedure that slows down the work. These fractions found to contain one or more components often require a second chromatography step to properly separate the components.
In an exemplary liquid chromatography system 80, signals 31 and 61 from detector 13 (eg ELSD) and UV detector 16 are sent to fraction collector 14, respectively, such as for collection of new sample fractions. Start some activities from the fraction collector 14. In a preferred embodiment, depending on (i) detector ELSD13, (ii) UV detector 16, or (iii) one or more detector signals 31 and 61 from both ELSD13 and UV detector 16, The fraction collector 14 collects a new sample.
Similar to the exemplary liquid chromatography system 80, in the liquid chromatography system 60 shown in the exemplary FIG. 6, signals 311 and 611 from the ELSD 131 and UV detector 161 are sent to the fraction collector 14, respectively. And initiate some activity from the fraction collector 14, such as collecting new sample fractions. In a preferred embodiment, depending on (i) detector ELSD131, (ii) UV detector 161 or (iii) one or more detector signals 311 and 611 from both ELSD131 and UV detector 161. The fraction collector 14 collects a new sample.
As mentioned above, the UV detector 16 (or UV detector 161) has n sensors and is configured to observe the sample at n specific wavelengths over a portion of the entire UV absorption spectrum. To. In the liquid chromatography system 80 shown in the exemplary FIG. 8, (i) a single signal from either ELSD 13 or UV detector 16, (ii) two from ELSD 13 and UV detector 16 or it. Depending on the above signal, or (iii) a composite signal with two or more detection responses at two or more specific UV wavelengths (up to n detection responses), the fraction collector 14 is a new sample. To collect. Similarly, the liquid chromatography system 60 shown in exemplary FIG. 6 is: (i) a single signal from either ELSD131 or UV detector 161 and (ii) 2 from ELSD131 and UV detector 161. Depending on one or more signals, or (iii) a composite signal with two or more detection responses at two or more specific UV wavelengths (up to n detection responses), the fraction collector 14 , Collect new samples.
Further, in an exemplary liquid chromatography system 80, the UV detector 16 may be used to generate a detector signal (not shown), the detector signal being (1) (i) single. It arises from a single detection signal response from the sensor of, or (ii) n detector responses of n detectors greater than 1, and the detection signal is (2) at least one splitter pump 15, ELSD13. And sent to T section 12. Further, a detector signal (not shown) resulting from the detector response of the ELSD 13 may be sent to the UV detector 16 to change one or more settings of the UV detector 16. Similarly, in the liquid chromatography system 60 shown in the exemplary FIG. 6, the UV detector 161 may be used to generate a detector signal (not shown), which is (1) ( i) results from a single detection signal response from a single sensor, or (ii) n detector responses from n detectors greater than 1, and the detector signal is (2) at least one. Sent to shuttle valve 151 and ELSD13. Further, a detector signal (not shown) resulting from the detection response of the ELSD 131 may be sent to the UV detector 161 to change one or more settings of the UV detector 161.
As shown in the exemplary liquid chromatography system 90 shown in FIG. 9, the location of different types of detectors within a given system is desired to provide one or more features of the system pro-contact. Can be adjusted as In an exemplary liquid chromatography system 90, the ELSD 13 is positioned downstream of the UV detector 16. In such a configuration, the UV detector 16 provides a detector response for the fraction collector 14 prior to the generation of the signal 31 from the ELSD 13 and the signal 61 (eg, (i) single). It is positioned so that it can generate a single detector response from a sensor or (ii) a signal resulting from n detector responses of n sensors greater than 1. The UV detector 16 also provides a detector response for at least one splitter pump 15, ELSD 13 and T section 12 so that the splitter pump 15, ELSD 13 and / or T section 12 can be driven or stopped. Generate a signal (not shown) (for example, (i) a single detector response from a single sensor, or (ii) a signal resulting from n detector responses from n sensors greater than 1). Positioned so that it can.
Although not shown, it is understood that the shuttle valve can be used in place of the T section 12 and splitter pump 15 in the exemplary liquid chromatography system 90 shown in FIG. 9 to provide similar system process features. I want to be. In such a configuration, the UV detector 16 provides a detector response for the fraction collector 14 prior to the generation of the signal 31 from the ELSD 13 and the signal 61 (eg, (i) single). Positioned so that a single detector response from a sensor, or (ii) a signal resulting from n detector responses from n sensors greater than 1) can be generated. The UV detector 16 also provides a detector response and a signal (not shown) (eg, (eg, (not shown)) so that the shuttle valve and / or ELSD can be driven or stopped for at least one shuttle valve and ELSD 13. It is positioned to be able to generate (i) a single detector response from a single sensor, or (ii) a signal resulting from n detector responses from n sensors greater than 1. Systems 60, 80, and 90 referred to ELSD and UV as detectors, but destructive detectors such as any EPD can be used for ELSD, and any non-destructive detector can be used. It can be used in place of the UV detector.
In another exemplary embodiment, the liquid chromatography system of the present invention is non-destructive with two or more non-destructive detectors (eg, one or more optical absorption detectors such as the UV detectors described above). A destructive system can be provided and there is no destructive detector (mass spectrometer) in the system. In one exemplary embodiment, the liquid chromatography system has two optical absorption detectors, such as a UV detector, and the method of analyzing the sample is to observe the sample at two or more specific wavelengths. Steps with two or more detectors; (i) changes in the first detector response at the first wavelength, (ii) changes in the second detector response at the second wavelength, or (iii) first detection It has a step of collecting a new sample fraction, depending on the change in the synthetic response represented by the instrument response and the second detector response.
In embodiments that use two or more optical absorption detectors, such as two or more UV detectors, the optical absorption detectors are given liquid chromatography systems to provide the benefits of one or more systems. Can be positioned within. Two or more optical absorption detectors can be positioned parallel to each other so that samples reach each detector at substantially the same time, and two or more optical absorption detectors can be positioned at substantially the same time. Generate signals (ie, first detector response and second detector response) and send the signals to the fraction collector.
In a further embodiment, a non-destructive detector (eg, RI detector, UV detector, etc.) alone or a destructive detector (eg, EPD, mass spectrometer, spectrophotometer, radiation spectrometer, NMR, etc.) ) Can be used in combination. For example, a destructive detector such as a mass spectrometer can simultaneously detect a peak of a component and the chemical properties associated with that peak. This enables quick determination of the fraction including the target. For other detection techniques, for example, a spectrophotometer, a mass spectrometer, a radiation spectrometer, an NMR, or the like requires a separation determination after the fraction containing the target. When two or more chemistries are simultaneously extracted from a flash cartridge (ie, they have the same residence time) and when a particular detector is used (ie, a detector that cannot identify differences in chemistries), these The detectors cannot determine the chemical composition, so they are deposited in the same vial by the system. In an exemplary embodiment in which the mass spectrum detector is used as a destructive detector, all components extracted simultaneously can be identified. This eliminates the need to check for purity after separation.
In any of the liquid chromatography systems described above, at least one detector, such as at least one UV detector, is downstream (eg, in series) of at least one UV detector or at least one other detector, such as ELSD. Positioning can be advantageous. In such an embodiment, the first detector response in the first detector provides a signal and (1) splitter pump, (2) shuttle valve, (3) second detector, and (4) T. Used to send a signal to at least one of the parts. For example, the first detector response of the first detector is used to generate a signal and send the signal to the splitter pump or shuttle valve, (i) drive the splitter pump or shuttle valve, and (ii) the splitter pump. Or stop the shuttle valve, (iii) change one or more flow or pressure settings of the splitter pump or shuttle valve, and at least one of (iv) or any combination of (i) to (iii). Do one. Appropriate flow and pressure settings include, but are not limited to, the flow and pressure settings described above. Typically, the signal is in the form of, for example, an electrical signal, a gas signal, a digital signal, a wireless signal.
In some embodiments, the plurality of detectors (ie, two or more detectors) are independent of the other detectors in the system, with each detector being (1) a splitter pump, (2) a shuttle valve. , (3) other detectors, (4) T section, positioned to be able to send signals to at least one of them. For example, multiple optical absorption detectors (eg, UV detectors) could be positioned within a given system to provide an independent signal to the shuttle valve, which was actively controlled by the shuttle valve. Have the fluid sample provided to another detector such as ELSD.
In other embodiments, the first detection response of the first detector is used to generate a signal and send the signal to the second detector, (i) driving the second detector, and (ii) second. 1 Drive the second detector at a wavelength substantially similar to the first wavelength used by the detector, and (iii) drive the second detector at a wavelength other than the first wavelength used by the first detector. (iv) Stop the second detector, (v) change some other settings of the second detector (for example, the observed wavelength of the second detector), or (vi) (i) Perform any combination from (v) to (v).
In other embodiments, the first detection response of the first detector can be used to generate a signal and send the signal to the T section, (i) open a valve, or (ii) a liquid chromatography system. Close the valve to start or stop the flow of fluid through a portion of the. As mentioned above, the signal is typically in the form of, for example, an electrical signal, a gas signal, a digital signal, or a wireless signal. C. Signal generation from detector response One method according to the invention can further include the step of generating a signal from one or more detector responses. In some embodiments, such as the exemplary liquid chromatography system 10 shown in FIG. 1, a single detector detects the presence of a sample component and is based on the presence and concentration of the sample component in the fluid stream. Generates a detector response. In other embodiments, such as the exemplary liquid chromatography system 50 shown in FIG. 6, two or more detectors can be used to detect the presence of one or more sample components and fluid flow. Two or more detection responses can be generated based on the presence and concentration of one or more sample components within.
As mentioned above, a given detector provides one or more detector responses, which generate a signal and one or more in a liquid chromatography system as described herein. It can be used to signal a component (eg, a fraction collector, other detector, splitter pump, shuttle valve, or T section). Typically, a change in a given detector response triggers the generation and transmission of a signal. Changes in a given detection response that can generate a signal and trigger transmission of a signal to one or more components are, but are not limited to, changes in the detector response value, to the threshold of the detector response value. Reached or exceeded, the time gradient of the detector response value, the threshold of the time gradient of the detector response value, the change of the time gradient of the detector response value, the change of the threshold value of the time gradient of the detector response value, or these Including combinations.
In one exemplary embodiment, the method of the invention comprises the step of generating a detector signal from at least one detector, which is (i) a gradient of the detector response as a function of time. (Ie, the first derivative of the detector response), (ii) the change in the gradient of the detector response as a function of time (ie, the second derivative of the detector response), (iii) optionally, of the detector response value. Generated in response to a threshold, or any combination from (iv) (i) to (iii) and a desired combination including at least (i) or at least (ii). In this exemplary embodiment, the substance is recognized from the shape of the detector response, and in particular from the first and / or second derivative (ie, tilt and tilt changes, respectively) with respect to the time of the detector response. .. In particular, the computer program analyzes the time sequence of the detector response values and evaluates their rate of change (ie, first derivative) and rate of change (ie, second derivative). If both the first and second derivatives are increasing, the substance is beginning to be detected. Similarly, when both the first derivative and the second derivative are decreasing, the detection of the substance is finished.
Actual detector values are typically noisy (eg jagged) and it is desirable to use a numerical lowpass filter for time (eg smoothing). Therefore, the step of generating a detector signal from at least one detector is preferably further (i) time-slope data, (ii) time-slope change, (iii) optionally, a threshold of detector response, or (iv) For any combination from (i) to (iii), it has steps to perform a numerical low-pass filter, (i) time slope, (ii) change in time slope, (iii) optionally detected. Identify the actual change in the threshold of the instrument response value, or any combination of (iv) (i) to (iii), from the possible noise of the detector response. In a preferred embodiment, a finite impulse response (FIR) filter or an infinite impulse response (IIR) filter can be used as a numerical lowpass filter for data on time (eg, a simple average of several samples). Typically, the deterministic algorithm uses a small number of sequential calls in time to confirm the actual detector response / signal rather than noise.
In another embodiment, the method of analyzing a sample comprises a step of generating a composite signal having a detection response component from each detector and a step of collecting a new sample fraction in response to changes in the composite signal. Can be done. In these embodiments, the steps to generate the composite signal are (i) the detection response value, (ii) the slope of the given detection response as a function of time (ie, the first derivative of the given detection response). , (Iii) obtained as a function of time R other changes in the slope of the detection response (ie, the second derivative of the given detection response), or (iv) each detector (ie each of two or more detectors) It can include a step of numerically correcting any combination from (i) to (iii). For example, in some embodiments, the composite signal is (i) the product of the detector response values of each detector (ie, each of two or more detectors) at a given time, (ii) detection at a given time. It can have the product of the first derivative of the instrument response, (iii) the product of the second derivative of the detector response at a given time, or any combination of (iv) (i) to (iii).
In other embodiments where a composite signal is used, the steps to generate the composite signal are (i) the detector response value, (ii) the gradient of the detector response given as a function of time (ie, the given detector). First derivative of the response), (iii) change in slope of the detector response given as a function of time (ie, second derivative of the given detector response), or (iv) within one detector Any combination from (i) to (iii) from each sensor (ie, n sensors observing the sample at n wavelengths), or (iv) with other detector responses present in the system. Any combination of, can have a step of numerically correcting. For example, in some embodiments, the composite signal is (i) the detector of each sensor in one detector (ie, n sensors observing the sample at n wavelengths) at a given time. The product of the response values, and the product of any additional detector response values from other detectors (eg, ELSD used in combination with the UV detector), (ii) one at a given time. The product of the first-order differentials of the detector response of each sensor in the detector (ie, n sensors observing the sample at n wavelengths), and any additional detection response from other detectors, (iii) The detector response of each sensor in one detector (ie, n sensors observing the sample at n wavelengths) at a given time, and optionally additions from other detectors. Can have a detection response, a product of second-order differentials, or any combination of (iv) (i) to (iii).
D. Collection of one or more sample fractions In order to collect one or more sample fractions in response to one or more signals from at least one detector in a given liquid chromatography system, the methods of the invention further include. Fraction collectors such as the exemplary fraction collector 14 shown in FIGS. 1-3A and 4-9 may be used. For example, in the exemplary liquid chromatography systems 10, 20, and 30, respectively shown in FIGS. 1, 2, and 3A, the method of analyzing a sample is further based on one or more signals from the first detector 13. A step of collecting one or more sample fractions can be provided. In the exemplary liquid chromatography systems 40, 50, 60 shown in FIGS. 4, 5 and 6, respectively, the method of analyzing the sample is further described in First Detector 13 (or First Detector 131), Second Detector. In response to one or more signals from 16 (or the second detector 161), or both the first detector 13 and the second detector 16 (or both the first detector 131 and the second detector 161). , Can be provided with a step of collecting one or more sample fractions.
In some embodiments of the invention, the fraction collector is configured to recognize, receive, and process one or more signals from at least one detector, and is also based on one or more signals. It is configured to collect one or more sample fractions. In other embodiments, an additional computer or microprocessor device is used to process one or more signals from at least one detector and then provide the fraction collector with a recognizable signal. The signal to the fraction collector is that the fraction collector is instructed to collect one or more sample fractions based on one or more signals from an additional computer or microprocessor device. Is.
As mentioned above, the system components can be positioned within a given liquid chromatography system in order to be able to provide one or more system features. For example, at least one detector has a time delay between (i) detecting a given detector response and (ii) collecting a sample fraction based on the signal generated from the detector response. Can be positioned within a given liquid chromatography system to minimize. In an exemplary embodiment of the invention, the liquid chromatography system is preferably of less than 2.0 seconds (or less than 1.5 seconds, less than 1.0 seconds, or less than 0.5 seconds) of a given detector signal. The time delay to the fraction collector (ie, the time delay between (i) the detection of a given detector and (ii) the step of collecting a sample fraction based on the signal generated by the detector). It has a maximum value.
In an exemplary embodiment of the invention using at least one detector (described below) with two or more detectors or n sensors, the liquid chromatography system is preferably less than 2.0 seconds (described below). Or less than 1.5 seconds, less than 1.0 seconds, or less than 0.5 seconds, time delay from any detector to the fraction collector of any detector signal (ie, (i) given detector Indicates the maximum value between the detection of the response and (ii) the time delay between the step of collecting the sample fraction based on the signal generated from the detector response (eg, single or compound signal).
E. Sample component separation step The method of the present invention uses a liquid chromatography (LC) step to separate the components in a given sample. Various LC columns, moving phases, and other process step conditions (eg, feed rate, gradient, etc.) can be used, depending on the particular sample.
Many LC columns can be used in the present invention. In general, polymer or inorganic normal phases, opposite phases, ion exchanges, affinities, hydrophobic interactions, hydrophilic interactions, mixed modes, and dimension exclusion columns can be used in the present invention. Exemplary commercially available columns are, but are not limited to, VYDAC®, GRACERESOLV®, DAVISIL®, ALLTIMA®, VISION® from Grace Davison Discovery Sciences. ), GRACEPURE , EVEREST , and DENALI , including those available from other similar companies.
In the present invention, many movable phase components can be used. Suitable mobile phase components are, but are not limited to, acetonitrile, dichloromethane, ethyl acetate, heptane, acetone, ethyl ether, tetrahydrofuran, chloroform, hexane, methanol, isopropyl alcohol, water, ethanol, buffers, and combinations thereof. including.
F. User interface step The method of analyzing a sample in the present invention can further comprise one or more steps in which an operator or user accesses one or more system components of a liquid chromatography system. For example, a method of analyzing a sample can include one or more of the following steps: That is, the step of placing the sample in a liquid chromatography system for testing; the step of adjusting one or more settings (eg, flow or pressure settings, wavelength, etc.) of one or more elements in the system; one or more. Steps to program at least one detector to generate a signal based on the desired mathematical algorithm that considers one or more detector responses from the sensor and / or multiple detectors; The step of programming one or more system components (other than the detector) to generate a signal based on the desired mathematical algorithm that considers the detector response; the signal from at least one detector (eg, a single signal). Steps to program a fraction collector to recognize (or a composite signal) and collect one or more sample fractions based on the received signal; the fraction collector is one or more system components (fractions). Recognize the input signal from at least one detector so that one or more samples can be collected based on the input from (other than the collector), and turn this input signal into a signal that the fraction collector can recognize and process. The step of programming one or more system components (other than the fraction collector) to convert; and other actions at the desired time or within the liquid chromatography system (eg, displayed to the operator or user). A step of driving or stopping one or more system components (eg, a T-valve, a splitter pump, a shuttle valve, or a detector) in response to a detected detector response can be provided.
II. Equipment for analyzing samples The present invention also relates to devices and equipment components capable of analyzing or contributing to the analysis of a sample using one or more of the steps of the method described above.
As mentioned above, in some exemplary embodiments of the invention, the device for analyzing the sample is T comprising (i) a chromatography column, (ii) a first inlet, a first outlet, and a second outlet. Part, (iii) Fraction collector that communicates fluid with the first outlet of part T, (iv) First detector that communicates fluid with the second outlet of part T, and (v) Exit 2 and second of part T The configuration can be provided to actively control the fluid flow to the first detector, which is positioned to communicate with the detector. In another exemplary embodiment of the invention, a shuttle valve can be used instead of the T section / splitter pump combination to actively control the fluid flow to the first detector.
Although not shown in Figure 1-9, any device described above (eg, an exemplary liquid chromatography system 10-90) or device component can further include system hardware, this system hard wafer. Is (i) recognition of a change in the detector response value or the detector response value, (ii) generation of a signal from the change in the detector response value or the detector response value, (iii) the signal to one or more system components. Allows transmission, (iv) recognition of the signal generated by the receiving component, (v) processing of the recognized signal within the receiving component, and (vi) initiation of processing steps of the receiving component in response to the recognized signal. To.
In one embodiment, the device (eg, liquid chromatography system 10-90) or a given device component can further include system hardware, the system hard wafer being the first detector, the splitter pump. Or allow the drive signal to be transmitted to the shuttle valve, (i) drive the splitter pump or shuttle valve, (ii) stop the splitter pump or shuttle valve, (iii) flow or pressure of the splitter pump or shuttle valve. Change one or more of the settings, or make any combination of (iv) (i) through (iii). Appropriate flow and pressure settings are, but are not limited to, from (i) valve position, (ii) splitter pump or shuttle valve pressure, (iii) air pressure to the valve, or (iv) (i). Includes any combination up to (iii).
In some embodiments, the splitter pump can be positioned between the T section and the first detector (eg, the splitter positioned between the T section 12 and the first detector 13 in FIG. 1). See Pump 15). In other embodiments, the first detector can be positioned between the T section and the splitter pump (eg, the first detector positioned between the T section 12 and the splitter pump 15 in FIG. 2). Please refer to).
In other embodiments, the apparatus of the present invention comprises (i) a chromatography column, (ii) two or more detectors, and (iii) a fraction collector that communicates fluid with two or more detectors. A fraction collector is configured to collect one or more sample fractions in response to one or more detector signals from two or more detectors. In some embodiments, the two or more detectors have two or more non-destructive detectors (eg, two or more UV detectors) and no destructive detector in the system.
If more than one detector is present, a splitter pump or shuttle valve can be used to split the volume of fluid flow between the first and second detectors. In other embodiments, a splitter pump or shuttle valve can be used to start or stop the fluid flow to the other detector in response to the detector response from one detector. In addition, multiple splitter pumps and / or shuttle valves can be used in a given system to actively control fluid flow to more than one detector.
As mentioned above, the device can further include system hardware, which allows the generation of detector signals from one or more detector responses. In one exemplary embodiment, the device comprises system hardware that allows the generation of a detector signal, which (i) tilts the detector response as a function of time. (Ie, first-order differentiation of detector response), (ii) change in slope of detector response as a function of time (ie, second-order differentiation of detector response), (iii) optionally threshold of detector response value , Or (iv) any combination of (i) to (iii), including at least (i) or (ii). The system hard wafer preferably also has low-pass numerical filtering capability, from (i) tilt data, (ii) tilt data changes, (iii) optionally, detector response thresholds, or (i). Any combination up to (iii), filtered over time, from possible noise in a given detector response, i) tilt data, (ii) changes in tilt data, (iii) optionally, detector Distinguish the actual changes in the response value threshold, or any combination from (i) to (iii).
In a multi-detector system, the system hard wafer allows the generation of a composite signal, including a detector response component from each detector, a detection response component from multiple sensors in a given detector. Can be used. In these embodiments, the system hardware is configured to send a fraction collector a command / signal instructing the fraction collector to collect a new sample fraction in response to changes in the composite signal. .. The composite signal is (i) the detector response value from each detector, (ii) the slope of the detector response given as a function of time (ie, the first derivative of the given detector response), (iii) time. Mathematical between changes in the slope of a given detector response (ie, the second derivative of a given detector response), or any combination of (iv) (i) to (iii), as a function of Correlation can be provided. For example, the composite signal is (i) the product of the detector response values of each detector at a given time, (ii) the product of the first derivative of the detector response values at a given time, (iii) given. It can include the product of the second derivative of the detector response in time, or any combination of (iv) (i) to (iii).
In one desirable configuration, the device for analyzing the sample has at least one detector configured to observe the sample at two or more specific optical wavelengths (eg, within the UV spectrum). In addition, the device has system hardware, and the system hardware wafer has a fraction collector that (i) changes in the detector response at the first wavelength, (ii) changes in the detector response at the second wavelength, or (iii) It is possible to collect new sample fractions in response to changes in the composite signal represented by the detector response at the first and second wavelengths. Each detector can operate at the same wavelength, at different wavelengths, or at multiple wavelengths. In addition, each detector can be in parallel, in series, or in some combination of parallel and series with each other.
As mentioned above, in one exemplary embodiment, the device samples at n optical wavelengths over part or all of the UV absorption spectrum (or any part of the absorption spectrum with other types of detectors). A single detector with n sensors configured for observation can be provided. Also, the device can be equipped with system hardware, where the fraction collector is (i) any one change in the response of n detectors at n optical wavelengths, (ii) n. Allows collection of new sample fractions in response to changes in the composite response, represented by the individual detector responses.
If a splitter pump or shuttle valve is present to actively control the flow of fluid to at least one detector, the device for analyzing the sample also produces drive vibrations to the splitter pump or shuttle valve. It can (i) drive the splitter pump or shuttle valve, (ii) stop the splitter pump or shuttle valve, (iii) change the flow rate or pressure of one or more of the splitter pump or shuttle valve, or (i) iv) Perform any combination of (i) to (iii). The drive signal can be generated by a system component such as a system operator or detector (ie, the drive signal depends on the detector response value or the detector response of the detector, as described above. Generated and transmitted by the detector as the value changes).
In a further embodiment of the invention, the apparatus for analyzing a fluid sample using a chromatography system is a first fluid passage flowing out of a chromatography column or cartridge; at least one capable of analyzing the fluid sample. Includes a detector; and a shuttle valve that moves a sample alicot of the fluid from the first fluid passage to the detector without substantially affecting the flow characteristics of the fluid through the first fluid passage. Since the first fluid passage or channel is substantially linear or straight, at least as it passes through a portion of the valve, the flow of fluid through the first fluid passage can be substantially laminar. In a further exemplary embodiment, the pressure of the fluid through the first fluid passage is substantially constant and / or does not increase substantially. In other embodiments, the fluid flow rate is substantially constant as it passes through the first fluid passage. In one alternative embodiment, a second fluid passage is used to transport an aliquot sample of fluid from the shuttle valve to the detector. Since the second fluid passage or channel is substantially linear or straight, at least through a portion of the valve, the flow through the second fluid passage can be substantially laminar. In one exemplary embodiment, the pressure of the fluid through the second fluid passage is substantially constant and / or does not increase substantially. In a further embodiment, the flow rate of the fluid can be substantially constant as it passes through the second fluid passage.
In a further exemplary embodiment, the apparatus for analyzing a fluid sample using chromatography is a first fluid passage flowing out of the chromatography column; the fluid to at least one detector capable of analyzing the sample. Second fluid passage for transporting samples; a shuttle valve that moves an aliquot sample of fluid from the first fluid passage to the second fluid passage and maintains a continuous second fluid passage through the shuttle valve. including. In one embodiment, a continuous first flow passage through the shuttle valve is maintained as the fluid aliquot sample is removed from the first fluid passage. In another embodiment, the continuous first and second flow passages through the shuttle valve are maintained as the fluid aliquot sample is removed from the first fluid passage and moved to the second flow passage.
In an exemplary embodiment of the invention, the apparatus for analyzing a sample further comprises a fraction collector configured to collect one or more sample fractions in response to one or more detector signals. Have detector signals: (i) 1st detector, (ii) 2nd detector (or any number of additional detectors), or () 1st and 2nd detectors (or optional) Emitted from both of the number of additional detectors). When multiple detectors are used, the device collects a fraction collector as described above in response to changes in the composite signal due to one or more detector responses from each detector. Can be configured to:
As mentioned above, in some exemplary embodiments, the device for analyzing the sample is a fraction collector configured to recognize, receive, and process the signal from at least one detector. And the fraction collector collects one or more sample fractions based on one or more signals. In other embodiments, the device that analyzes the sample has an additional computer or microprocessor device that processes one or more signals from at least one detector and fractionally collects the input signal. Can be converted into a recognizable signal. In this latter embodiment, the fraction collector collects one or more sample fractions based on one or more signals from an additional computer or microprocessor device and becomes the signal processing component of the fraction collector. Not based.
Any of the exemplary liquid chromatography systems described above can be equipped with any number of detectors, splitter pumps, T-sections, shuttle valves, which are provided to provide one or more system performances. Strategically placed within the system. For example, although not shown in the exemplary liquid chromatography system 60 of FIG. 6, additional detectors are provided between column 11 and shuttle valve 151 and / or between shuttle valve 151 and detector 161. Can be positioned to. Although not shown in the exemplary liquid chromatography system 70 shown in FIG. 7, additional detectors are provided between column 11 and shuttle valve 151 and / or between shuttle valve 151 and shuttle valve 171. , And / or can be positioned between the shuttle valve 171 and the fraction collector 14. Additional detectors can be similarly positioned within the exemplary liquid chromatography systems 80 and 90 shown in FIGS. 8 and 9, respectively.
As described below, a number of commercially available components can be used in the apparatus of the present invention.
A. Chromatography column Any known chromatography column can be used in the apparatus of the present invention. Suitable commercially available chromatography columns are, but are not limited to, from Grace Davison Discovery Sciences, Deerfield, Illinois, GRACEPURE , GRACERESOLV , VYDAC , and Available under the DAVISIL® trademark.
B. Detector Any known detector can be used in the apparatus of the present invention. Suitable commercially available detectors are, but not limited to, UV detectors available under the USB2000 trademark from Ocean Optics (Dannydin, Florida); Grace Davison Discovery Sciences (Dearfield). Evaporative Light Scattering Detectors (ELSDs), available from Illinois, USA under the trademark 3300 ELSD; Aggregate Nucleating Light Scattering Detectors (CNLSDs), available under the QT-500 trademark from Quant (Brain, Mass.); CORONA CAD trademark from ESA (Chelmsford, Mass.) Corona Discharge Detectors (CDDs) available at; Refractive Rate Detectors (RIDs) available at 2414 from Waters Corporation (Milford, Mass.); And Laboriance (St. Includes fluorescence detectors (FDs) available under the ULTRAFLOR trademark from Collect (Pennsylvania).
In some embodiments, commercially available detectors may need to be modified or programmed. Alternatively, some detectors may need to be constructed to perform one or more of the steps of the method described above in the present invention.
C. Splitter pump Any known splitter pump can be used in the apparatus of the present invention. Suitable commercially available splitter pumps include, but are not limited to, splitter pumps available under the LIQUID MICRO trademark from KNF (Trenton, NJ).
D. Shuttle valve Any known shuttle valve can be used in the apparatus of the present invention. Suitable commercially available shuttle valves are, but are not limited to, CHEMINERT from Valco (Texas, Houston), shuttle valves available under the Rheodyne trademark, and MRA from Idex Corporation. Includes shuttle valves available under the Trademark (Registered Trademark) and continuous flow shuttle valves described herein.
E. Fraction collector Any known fraction collector can be used in the apparatus of the present invention. Suitable commercially available fraction collectors include, but are not limited to, fraction collectors available from Gilson (Middleton, Wisconsin) with a label of 215.
In some embodiments, commercially available fraction collectors may need to be modified and / or programmed. Alternatively, some fraction collector may need to be constructed to perform one or more of the steps of the method described above in the present invention. For example, a fraction configured to recognize, receive, and process one or more signals from at least one detector to collect one or more sample fractions based on the one or more signals. Collectors may not be commercially available at this time.
III. Computer software The invention further relates to a computer-readable medium containing computer-executable instructions for performing one or more of the steps of the method described above. For example, a computer-readable medium containing computer-executable instructions adjusts one or more settings (eg, flow settings, wavelength, etc.) of one or more elements in the system; one or more. Generate a signal based on the desired mathematical algorithm that considers the detector response; recognize the signal from at least one detector; collect one or more sample fractions based on the received signal; at least one It recognizes the input signal from the detector and converts the input signal into a signal that the fraction collector can recognize and process, and the fraction collector can do it at the desired time or at some other time in the liquid chromatography system. Collect one or more sample fractions based on input from one or more system components (eg T-valve, splitter pump, shuttle valve, or detector) depending on the activity (eg, detector response). Store instructions to allow you to.
IV. Application / Use The methods, devices and computer software described above can be used to detect the presence of one or more components in various samples. The methods, devices, and computer software described above can be applied to any industry that uses liquid chromatography, including, but not limited to, the petroleum industry, the pharmaceutical industry, and analytical research fields.
Example The present invention is further described by the following examples. These examples do not limit the scope of the invention in any way. By referring to the description herein, various other embodiments, modifications, and equivalents will be clearly recognized by those skilled in the art without departing from the spirit and / or the appended claims. Will be done.
<u style="single">Example 1</u>In this example, a flash REVELERIS system (available from Grace Vavison Discovery Sciences) was used. A 4 mL mixture containing sucrose and aspirin was injected into a 4 g GRACE RESOLV C18 flash column (available from Grace Vavison Discovery Sciences) mounted within the flash system. A 50/50 methanol / water mobile phase was pumped through the system using the ALLTECH® model 300LC pump. The column effluent was directed to the KNF splitter pump, which led the column effluent to ALLTECH® 3300 ELSD at 300 uL / min. The rest of the effluent flowed through the Ocean Optics UV detector to the Gilson fraction collector.
Sucrose and aspirin were separated on a flash column. Both sucrose and aspirin were detected by ELSD. Only UV detectors detected aspirin. Both detectors responded to aspirin at the same time. Fraction collectors deposited sucrose and aspirin in separate collection vials in response to composite signals from UV and ELSD detectors.
<u style="single">Example 2</u>In this example, a flash REVELERIS system (available from Grace Vavison Discovery Sciences) was used. A 4 mL mixture containing dioctylphthalate and butylparaben was injected into a 4 g GRACE RESOLV C18 flash cartridge (available from Grace Vavison Discovery Sciences) mounted within the flash system. An 80/20 methanol / water mobile phase was pumped using an ALLTECH® model 300LC pump. The column effluent was guided to the shuttle valve described herein, which led the column effluent to ALLTECH® 3300 ELSD at 300 uL / min. The rest of the effluent flowed through the Ocean Optics UV detector to the Gilson fraction collector.
The mixture of these two components is a non-color-developing compound (a substance that does not absorb UV light) and a color-developing compound. The non-color-developing compound is first extracted from the flash cartridge. FIG. 11 shows a chromatogram, which shows that only ELSD identifies all components in the sample, evidenced by two peaks in the chromatogram. The UV detector does not identify non-coloring compounds at the two wavelengths (identified as the first peak in ELSD). Only ELSD signals will be able to properly control the fraction collector and capture both compounds. When a UV detector drives a fraction collector (as in a traditional flash system), the first compound is sent to the disposal unit and improperly deposited in the collection vessel, and these fractions. Is not found to contain the desired sample. In conventional flash equipment, all fractions are screened by thin layer chromatography (TLC) after chromatographic separation to look for substances that the UV detector could not identify. This example shows that according to the present invention, a flash device equipped with ELSD can identify and separate both color-developing and non-color-developing compounds without the need for post-separation RLC screening.
Although the present invention has been described with a limited number of embodiments, these particular embodiments are not intended to limit the scope of the invention, and the scope of the invention is described in the appended claims. It is described in the range of. By referring to the exemplary embodiments herein, one of ordinary skill in the art will recognize that further modifications, equivalents, and variations are possible. All parts in the example, percentages, like the rest of the specification, are by weight unless otherwise stated. In addition, the range of numbers stated in the specification or claims to indicate a particular property, unit of measure, condition, physical condition, proportion is intended to be literally and explicitly integrated by reference. The numbers in the range include any subset within the stated range. For example, when a numerical range with a lower bound RL and an upper bound RU is disclosed, any number R within this range is explicitly disclosed. In particular, numerical values R in the following range are specifically disclosed. That is, R = RL + k (RU-RL) is disclosed, where k is a variable in the range of 1% to 100% in 1% increments, for example, k is 1%, 2%, 3%, 4%, 5% ... 50%, 51%, 52% ... 95%, 96%, 97%, 98%, 99%, or 100%. In addition, any numerical range indicated by any two values of R, calculated as described above, is disclosed. In addition to these disclosures, any modifications of the invention will be apparent to those skilled in the art from the above description and accompanying drawings. Such amendments are intended to be within the appended claims. All references cited herein are integrated herein.
Every citation, both ways
| Document | Relation | Office | Cited during |
|---|---|---|---|
| JP2014514541A | Cited by | Japan | Examiner |
| JP2004045263A | Cites | Japan | Search report |
| JP2004045263A | Cites | Japan | Examiner |
| JP2007183173A | Cites | Japan | Examiner |
| JP2007183173A | Cites | Japan | Search report |
| JPH0329850A | Cites | Japan | Examiner |
| JPH0329850A | Cites | Japan | Search report |
| JPH04326058A | Cites | Japan | Search report |
| JPH06118073A | Cites | Japan | Search report |
| JPH0643075A | Cites | Japan | Search report |
| JPH11258224A | Cites | Japan | Search report |
| JPS6011172A | Cites | Japan | Examiner |
| JPS6011172A | Cites | Japan | Search report |
60 members in 15 offices
Priority claims9
| Document | Office | Kind | Date |
|---|---|---|---|
| 559007 | United States of America | P | |
| 559007 | United States of America | P | |
| 61005590 | United States of America | – | |
| 2008013359 | United States of America | W | |
| 2008013359 | United States of America | W | |
| 2007005590 | – | – | – |
| 2008013359 | – | – | – |
| US20070005590P | – | – | – |
| WO2008US13359 | – | – | – |
Members60
| Document | Office | Kind | |
|---|---|---|---|
| AU2008336091A1 | Australia | A1 | |
| CA2684774A1 | Canada | A1 | |
| WO2009075764A2 | World Intellectual Property Organization (WIPO) | A2 | |
| MX2009013107A | Mexico | A | |
| AR069597A1 | Argentina | A1 | |
| TW201022666A | Taiwan Province of China | A | |
| WO2009075764A3 | World Intellectual Property Organization (WIPO) | A3 | |
| NO20100961L | Norway | L | |
| US2010238444A1 | United States of America | A1 | |
| CN101849184A | China | A | |
| EP2232253A2 | European Patent Office (EPO) | A2 | |
| KR20100108331A | Republic of Korea | A | |
| JP2010540965AThis record | Japan | A | |
| RU2009137790A | Russian Federation | A | |
| HK1149078A1 | Hong Kong, China | A1 | |
| US2011301865A1 | United States of America | A1 | |
| US2011301868A1 | United States of America | A1 | |
| US2011310390A1 | United States of America | A1 | |
| US8115930B2 | United States of America | B2 | |
| US8305581B2 | United States of America | B2 | |
| US8305582B2 | United States of America | B2 | |
| US8314934B2 | United States of America | B2 | |
| JP2013011615A | Japan | A | |
| JP2013011616A | Japan | A | |
| US2013074579A1 | United States of America | A1 | |
| AU2013203554A1 | Australia | A1 | |
| AU2013203558A1 | Australia | A1 | |
| AU2008336091B2 | Australia | B2 | |
| KR20130083490A | Republic of Korea | A | |
| AU2013228049A1 | Australia | A1 | |
| KR20130119504A | Republic of Korea | A | |
| JP2013231734A | Japan | A | |
| JP2014038105A | Japan | A | |
| JP2014038106A | Japan | A | |
| CN103630639A | China | A | |
| CN103630640A | China | A | |
| CN103630641A | China | A | |
| CN103630642A | China | A | |
| CN103713073A | China | A | |
| CN101849184B | China | B | |
| KR20140114071A | Republic of Korea | A | |
| BRPI0811525A2 | Brazil | A2 | |
| EP2803994A1 | European Patent Office (EPO) | A1 | |
| JP2015038505A | Japan | A | |
| AU2013203558B2 | Australia | B2 | |
| AU2013228049B2 | Australia | B2 | |
| KR20150070427A | Republic of Korea | A | |
| KR20150079988A | Republic of Korea | A | |
| JP2015135346A | Japan | A | |
| KR20150090283A | Republic of Korea | A | |
| HK1204676A1 | Hong Kong, China | A1 | |
| KR20160013277A | Republic of Korea | A | |
| US2016069846A1 | United States of America | A1 | |
| US9322813B2 | United States of America | B2 | |
| KR101626272B1 | Republic of Korea | B1 | |
| CN103630640B | China | B | |
| CN103630642B | China | B | |
| JP5973522B2 | Japan | B2 | |
| CN103713073B | China | B | |
| JP6052507B2 | Japan | B2 |
11 legal events, as the office reported them to INPADOC
Over the term
Point at a mark for the eventEvents
| Event | Code | |
|---|---|---|
| Decision of refusalJAPANESE INTERMEDIATE CODE: A02A02 | A02 | |
| Written permission of extension of timeJAPANESE INTERMEDIATE CODE: A602A602 | A602 | |
| Written request for extension of timeJAPANESE INTERMEDIATE CODE: A601A601 | A601 | |
| Notification of reasons for refusalJAPANESE INTERMEDIATE CODE: A131A131 | A131 | |
| Written amendmentJAPANESE INTERMEDIATE CODE: A523A521 | A521 | |
| Written permission of extension of timeJAPANESE INTERMEDIATE CODE: A602A602 | A602 | |
| Written request for extension of timeJAPANESE INTERMEDIATE CODE: A601A601 | A601 | |
| Written permission of extension of timeJAPANESE INTERMEDIATE CODE: A602A602 | A602 | |
| Written request for extension of timeJAPANESE INTERMEDIATE CODE: A601A601 | A601 | |
| Notification of reasons for refusalJAPANESE INTERMEDIATE CODE: A131A131 | A131 | |
| Report on retrievalJAPANESE INTERMEDIATE CODE: A971007A977 | A977 |
Numbers
- Publication
- 2010540965
- Publication, DOCDB
- 2010540965
- Publication, EPODOC
- JP2010540965
- Application
- 2010528007
- Application, DOCDB
- 2010528007
- Application, EPODOC
- JP20100528007
Titles2
- Japanese
- サンプルの画分を収集しサンプルを分析するための方法および装置
- English
- Methods and equipment for collecting sample fractions and analyzing samples
Classification
- CPC, 7
- G01N30/82
- G01N30/28
- G01N2030/621
- G01N30/74
- G01N30/78
- G01N2030/322
- G01N30/72
- IPC, 4
- G01N30 78
- G01N30 84
- G01N30 74
- G01N30 72
Designated states4
- Regional, 4
- Zimbabwe
- Turkmenistan
- Türkiye
- Togo