JP6063529B2 - 物質分離用前処理カートリッジ及びそれを利用した前処理方法 - Google Patents
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Description
1−a)アミノプロピルシリカゲルへ重合開始剤の導入するために下記の化合物を使用した。
アミノプロピルシリカゲル 5g
V−501[4,4’−アゾビス(4−シアノバレリン酸(4,4’−azobis(4−cyanovaleric acid)(分子量:280・28)] 3.5g(12.5mmol)
EEDQ[N−エトキシカルボニル−2−エトキシ−1,2−ジヒドロキノリン(N−Ethoxycarbonyl−2−ethoxy−1,2−dihydroquinoline)(分子量:247.30)] 6.18g(25.0mmol)
それぞれのものを上述の割合で配合し、V−501に対しEEDQを縮合剤として使用し、これらをDMF 50mlに溶解させ、その中にアミノプロピルシリカゲルを分散させることで反応させた。これを遮光下で30分間N2(窒素)ガスでバブリングし、その後完全にN2置換し、N2風船をつけて室温で6時間反応させた。反応後、ろ過してからDMFで洗浄した。これによりシリカゲル表面に重合開始剤を導入した。
1−b)温度応答性固定相表面の形成のために下記の化合物を使用した。
1−aで作製したV−501結合シリカゲル4g
IPAAm 10g
BIS[N,N’−メチレン−ビス(アクリルアミド)(N,N’−Methylene−bis(acrylamide))(分子量:154.17)] 0.27g
上述の量で1−aで作製したV−501結合シリカゲル、IPAAm、BISをEtOH 200mlに溶解させた。これを遮光下で1時間N2バブリングした。その後、完全にN2置換し、N2風船をつけて70℃(油浴)で5時間反応させた。これにより正荷電を持つPIPAAm表面のゲル層を形成した。反応後、ろ過してからメタノールと水で洗浄した。これを充填剤として減圧乾燥してデシケーターに保存した。この充填剤150mgを使って、前処理用カートリッジ((4mm/1mL)を作製した。
Claims (9)
- 固定相表面に0〜80℃の温度範囲内で水和力が変化するポリマーが被覆され、そのポリマー鎖の温度変化による収縮、膨潤により当該固定相表面と分離したい物質との親和性を変えられる固定相が充填された、物質分離用前処理カートリッジであって、
該固定相が100μm以下の粒径の異なるシリカビーズ、及び/またはポリマービーズの混合物であり、
該ポリマーは、N−イソプロピルアクリルアミドの単独の重合体、もしくはN−イソプロピルアクリルアミドとその他のモノマーとの共重合体であり、
該ポリマーの固定相表面への被覆量は0.8〜10.0mg/m 2 であり、
該前処理カートリッジの素材は、ポリメチルメタクリル酸メチル(PMMA)、ポリカーボネート、ポリプロピレン、ポリエチレン、ポリメチルペンテン、ポリスチレン、ポリテトラフルオロエチレン、ABS樹脂、ポリジメチルシロキサン、シリコン、及びそれらの高分子化合物を含む共重合体あるいは複合体からなる群から選ばれ、
該前処理カートリッジは横断面が円の円筒形であり、流路の一端に試料の送液口を有し、及び流路の他端に試料の溶出口を有する、
前記物質分離用前処理カートリッジ。 - 前記その他のモノマーが疎水性モノマーである、請求項1に記載の物質分離用前処理カートリッジ。
- 前記N−イソプロピルアクリルアミド及び/またはその他のモノマーが荷電を有する、請求項1または2に記載の物質分離用前処理カートリッジ。
- 特定の温度のもとで利用し、水系移動相によって試料中の物質を分離することによって、試料液の前処理を行うための、請求項1〜3のいずれか1項に記載の物質分離用前処理カートリッジ。
- 前記試料の前処理が除タンパク質操作である、請求項4に記載の物質分離用前処理カートリッジ。
- 前記試料の前処理は、試料液中の分離したい物質を一度、前処理カートリッジの固定相表面に付着させ、その後、固定相表面の性質を変えることで溶出させるものである、請求項4または5に記載の物質分離用前処理カートリッジ。
- 前記試料が環境試料、血液、血漿、又は尿である、請求項4〜6のいずれか1項に記載の物質分離用前処理カートリッジ。
- 前記試料中の物質が、農薬、医薬品、ペプチド、又は蛋白質である、請求項4〜7のいずれか1項に記載の物質分離用前処理カートリッジ。
- 遠心力、注射器による加圧、又は吸引により前記試料を送液する、請求項1〜8のいずれか1項に記載の物質分離用前処理カートリッジ。
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JP6169885B2 (ja) * | 2013-05-07 | 2017-07-26 | 株式会社日立製作所 | 精製装置及び精製方法 |
US10359402B2 (en) * | 2015-03-25 | 2019-07-23 | Hitachi High-Tech Science Corporation | Two-dimensional liquid chromatographic analyzer and analytical method |
WO2016208777A1 (ja) * | 2015-06-26 | 2016-12-29 | 国立研究開発法人国立循環器病研究センター | 細胞培養器 |
EP3578972A1 (en) * | 2018-06-06 | 2019-12-11 | Blink AG | A device for fractionating a suspension sample |
JP7471626B2 (ja) | 2019-11-29 | 2024-04-22 | 慶應義塾 | 細胞精製用材料およびその利用 |
CN111330311B (zh) * | 2020-02-13 | 2021-02-23 | 浙江大学 | 一种相变诱导靶标富集的方法 |
CN113842672B (zh) * | 2021-09-16 | 2023-02-03 | 暨南大学 | 一种固相微萃取薄膜及其制备方法与应用 |
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JP3694519B2 (ja) | 2004-10-26 | 2005-09-14 | 松下電器産業株式会社 | ディスク記録再生装置 |
JP2008012383A (ja) | 2006-07-03 | 2008-01-24 | Hitachi Plant Technologies Ltd | 包括固定化担体、包括固定化担体を用いた廃水処理装置及び廃水処理方法 |
JP5635725B2 (ja) * | 2007-03-27 | 2014-12-03 | 株式会社セルシード | 生理活性高分子物質の分離方法 |
JP2009174938A (ja) * | 2008-01-23 | 2009-08-06 | Toray Ind Inc | プレカラム |
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2010
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- 2010-09-01 EP EP10813743.1A patent/EP2474827A4/en not_active Withdrawn
- 2010-09-01 WO PCT/JP2010/064956 patent/WO2011027794A1/ja active Application Filing
- 2010-09-01 JP JP2011529921A patent/JPWO2011027794A1/ja active Pending
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JP2015232573A (ja) | 2015-12-24 |
EP2474827A1 (en) | 2012-07-11 |
EP2474827A4 (en) | 2013-11-20 |
WO2011027794A1 (ja) | 2011-03-10 |
JPWO2011027794A1 (ja) | 2013-02-04 |
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