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Purification of anionic fluorescent probes through precise fraction collection with a two-point detection system using multiple-stacking preparative capillary transient isotachophoresis

2点検出システムを有するキャピラリー過渡的等速電気泳動法を用いた精確な分取法の構築と陰イオン性蛍光プローブの精製

原賀 智子   ; 辻村 大翔*; 宮内 さおり*; 上村 拓也*; 渋川 雅美*; 齋藤 伸吾*

Haraga, Tomoko; Tsujimura, Hiroto*; Miyauchi, Saori*; Kamimura, Takuya*; Shibukawa, Masami*; Saito, Shingo*

アクチノイド等の分析に必要な蛍光性試薬(陰イオン性蛍光プローブ)を高純度化するための技術を開発した。微小サンプルの分析に用いられるキャピラリー電気泳動法のうち、検出点を複数個所有する過渡的等速電気泳動法を開発し、キャピラリー中の泳動速度を泳動毎に正確に算出することにより、分取のタイミングを調整し、目的の成分のみを精確に分取する手法を確立した。これにより、分析に必要な試薬に含まれる不純物を除去し、95$$sim$$99%の高純度化に成功した。

A novel combination of CE-based separation techniques was used for the precise fractionation of ionic compounds from impurities. The combination of on-capillary concentration and separation using transient isotachophoresis, with multiple injections and a two-point detection system provided higher efficiency, and accuracy at a microliter-scale injection volume, than when CE was individually used for purification. In this paper, we present successful applications of the CE fractionation techniques for the purification of fluorescein, fluorescein-4-isothiocyanate, two fluorescent metal ion probes, and a fluorescein-modified DNA aptamer. The purity of the isolated fluorescent probes ranged from 95 to 99%. The purified probe solutions were practical for use as purified stock solutions. The method developed was useful for the purification of anionic fluorescent reagents to be of ultratrace analytical grade for use with CE-LIF.

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パーセンタイル:46.03

分野:Biochemical Research Methods

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