2 詳細
Development of hydrophilic fluorogenic derivatization reagents for thiols: 4-(N-acetylaminosulfonyl)-7-fluoro-2,1,3,-benzoxadiazole and 4-(N-trichloroacetylaminosulfonyl)-7-fluoro-2,1,3,-benzoxadiazole
K. Okabe, R. Wada, K. Ohno, S. Uchiyama, T. Santa and K. Imai
J. Chromatogr. A, 2002, 982, 111-118.
■ Abstract
Sensitive, reactive, and hydrophilic fluorogenic reagents for thiols with the benzofurazan skeleton, 4-(N-acetylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (AcABD-F) and 4-(N-trichloroacetylaminosulfonyl) -7-fluoro-2,1,3-benzoxadiazole (TCAcABD-F) have been developed.
These reagents reacted with thiols within 10 min at 60 degree Celsius.
AcABD-F and TCAcABD-F themselves do not fluoresce but are strongly fluorescent after the reaction with thiol compounds.
The generated derivatives were highly water-soluble, since they dissociated a proton and ionized in the neutral pH region.
The derivatives with four biologically important thiol compounds were separated on a reversed-phase HPLC column and detected fluorometrically at 504 nm with excitation at 388 nm.
The detection limit attained for homocysteine with AcABD-F was 25 fmol on column (11 nM) (S/N = 3), and that for glutathione with TCAcABD-F was 45 fmol on column (20 nM).
■ ひとこと
５年後輩に当たる岡部君が，４年後輩の和田さんのテーマを引き継いでまとめた結果です．
なぜかこのころ研究室では水溶性の試薬が一時的なブームになってました．ほとんど何もやっていないのですが，AcABD-F が水溶性試薬になるのではないか・・・と思いついたのが，僕だったのです．