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1. Moving affinity boundary electrophoresis and its selective isolation of histidine in urineElectronic supplementary information (ESI) available: Further experimental information and results. See DOI: 10.1039/c000472c.

Author

Jia MengThe first two authors contributed equally to the paper., Wei Zhang, Cheng-Xi Cao, Liu-Yin Fan, Jin Wu, and Qiu-Ling Wang

Subjects
*CAPILLARY electrophoresis, *URINALYSIS, *CHEMICAL affinity, *METAL ions, *METABOLITES, *LIGANDS (Biochemistry), *ANALYTICAL chemistry
Abstract

Affinity analysis is a key biotechnique used in the fields of biology and biomedicine. Herein, we advanced the concept of moving affinity boundary (MAB) using metal ion Ni(ii) and histidine (His) as the model inorganic ion and ligand, respectively, developed the simple method of MAB affinity capillary electrophoresis (MAB-ACE), and carried out the relative experiments. The experiments manifested that (a) an MAB could be created with the model metal ion and ligand; (b) the MAB-ACE could specifically capture His rather than other amino acids, or numerous metabolites in human urine; and (c) the capture had the merits of simultaneous focusing and separation to the target metabolite of His. It was further revealed that the specificity of MAB-ACE was originated from the selective affinity interaction and the effective control of affinity conditions. The analyses of His in raw urine by the MAB-ACE are in agreement with those viathe standard amino acid analyzer, indicating the reliability of the developed method. Additionally, the MAB-ACE with UV detector had good sensitivity (LOD = 43 ng mL−1, S/N = 3), 1.0–150 μM linearity and <5% intra-/inter-day variations. The novel method has an evident potential application for capture of a target metabolite in complex biological sample. [ABSTRACT FROM AUTHOR]

Published
2010
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2. キャピラリー電気泳動による誘導体化したチオール類の分離と検出

Author

Yukiko, Imaizumi, Tomoe, Yamashiro, yoshiko, Arikawa, 原著, Original Paper, 日本女子大学理学部物質生物学科, and Dept. of Chemical and Biological Sciences, Japan Women's University

Subjects
キャピラリー電気泳動, 誘導体化, capillary electrophoresis, Ellman試薬, Ellman's reagent, derivertized thiol, チオール類, thiols
Abstract

低分子のチオール類は揮発性が高くまた空気中で酸化されやすい。そこで5,5'-ジチオビス(2-ニトロ安息香酸; Ellman試薬)を用いてチオールを誘導体化したのち, キャピラリー電気泳動によりチオール類の分解・検出を行った。SH基を有する化合物にEllman試薬を過剰に加えると, 混合ジスルフィドとチオレートアニオンが生成するが, 電気泳動を行うことにより, 一部重複していた各成分のピークの分離が可能となった。泳動液は80μMテトラデシルアンモニウムブロミド(TTAB)を含む10mMリン酸二水素ナトリウム溶液を用いた。 TTAB溶液を添加することで, キャピラリー内壁の電荷を制御し電気浸遠流を減少させることにより各成分の持つ構造の違いによる泳動時間の差が明確になり分離が完全になった。キャピラリーは内径0.075mm, 全長500mm(有効長378mm)の溶融シリカ管を用いた。試料は落差10mmで30秒間キャピラリー内に注入し印加電圧十30kVにて泳動を行った。キャピラリー周囲は25℃に保った。検出にはEllman試薬を用いて誘導化したチオール類の吸収極大波長である357nmを用いた。エタンチオール, 1-プロパンチオール, 1-ブタンチオールを短時間で分離・検出できた。, The determination and detection of low molecular weight thiols by capillary electrophoresis (CE) system has been studied. Though electrophoresis of thiols has to be carried out at low pH due to their instability, each migration time of thiols was fairly long and the separation of thiols wasn't completely. We examined to derivatize thiols with 5,5'-dithiobis (2-nitorobenzoic acid) which has been used as the photometric reagent for the determination of thiols, usually called as Ellman's reagent. In this study, ethanethiol, 1-propanethiol and 1-butanethiol were investigated to be separated by CE with UV detection. Running buffer was prepared to be pH 7.4 containing 1OmM disodium hydrogenphosphate and 80μM tetradecyltrimethylammonium bromide. Ethanethiol, 1-propanethiol and 1-butanethiol were prepared as a 0.5mM solution in lOmM sodium phosphate buffer (pH=7.4). The applied voltage was set at +30kV. The detection of derivatized rhiols was measured at 357 nm. These thiols were separated each other in 18min.

Published
2003