title: Separation-oriented derivatization of native fluorescent compounds through fluorous labeling followed by liquid chromatography with fluorous-phase.
abstract: We have developed a new and simple method based on "fluorous derivatization" for LC of native fluorescent compounds. This method involves the use of a column with a fluorous stationary phase. Native fluorescent analytes with target functional groups are precolumn derivatized with a nonfluorescent fluorous tag, and the fluorous-labeled analytes are retained in the column, whereas underivatized substances are not. Only the retained fluorescent analytes are detected fluorometrically at appropriate retention times, and retained substrates without fluorophores are not detected. In this study, biologically important carboxylic acids (homovanillic acid, vanillylmandelic acid, and 5-hydroxyindoleacetic acid) and drugs (naproxen, felbinac, flurbiprofen, and etodolac) were used as model native fluorescent compounds. Experimental results indicate that the fluorous-phase column can selectively retain fluorous compounds including fluorous-labeled analytes on the basis of fluorous separation. We believe that separation-oriented derivatization presented here is the first step toward the introduction of fluorous derivatization in quantitative LC analysis.
authors.affiliation: Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Sakaguchi
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Yoshida
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Todoroki
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Nohta
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Yamaguchi
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000032:analysis / Q000097:blood; D002264:Carboxylic Acids / Q000032:analysis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D002264:Carboxylic Acids; D005456:Fluorescent Dyes