title: 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.
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 degrees C. 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) (signal-to-noise ratio = 3), and that for glutathione with TCAcABD-F was 45 fmol on column (20 nM).
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Kohki
authors.initials: K
authors.lastname: Okabe
authors.forename: Reiko
authors.initials: R
authors.lastname: Wada
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Ohno
authors.forename: Seiichi
authors.initials: S
authors.lastname: Uchiyama
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Santa
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Imai
mesh_terms: D001583:Benzoxazoles / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: C474129:4-(N-acetylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole; C474130:4-(N-trichloroacetylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole; D001583:Benzoxazoles; D005456:Fluorescent Dyes; D013438:Sulfhydryl Compounds