Summary
The use of 2-(9-carbazole)ethyl chloroformate (CEOC) for pre-column derivatization of biogenic amines (BA) has been tested for the first time. The reagent reacts completely with BA within 3 min at ambient temperature in acetonitrile solution to form stable derivatives that are readily analyzed by reversed-phase HPLC. Study of the derivatization conditions revealed derivatization yields to be excellent in borate buffer over the pH range 9.0–10.0. Maximum yields were obtained by use of a three- to fourfold molar excess of reagent. The reaction is extremely tolerant of common buffer salts, no decrease in reaction yield is discernible in well-buffered samples. The emission maximum for the CEOC-derivatives is 360 nm (λ ex = 293 nm). All the derivatives fluoresced strongly and direct injection of the reaction mixture was possible, with no significant disturbance from the major fluorescent reagent degradation by-products, 2-(9-carbazole)ethanol (CEOC-OH) and bis-(2-(9-carbazole)ethyl) carbonate (CEOC)2. Separation of the derivatized BA by high-performance liquid chromatography with gradient elution was tested on a Hypersil BDS C18 column. Excellent response linearity was observed over the concentration range from 0.25 to 94.6 μmol L−1 for the labeled BA. Detection limits were 117–840 fmol at a signal-to-noise ratio of 3∶1. Analysis of BA in a shrimp sauce extract was conducted to demonstrate the applicability of the technique to real sample matrixes; results were satisfactory.
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Vasundhara, T.S.; Kumudavally, K.V.; Jayathiilakan, K.; Jeyashakila, R.J. Food Sci. Technol. 1998,35, 551.
Kirschbaum, J.; Rebscher, K.; Brückner, H.J. Chromatogr. A 2000,881, 517.
Eclerq, M.; Gouilloux, L.Technol. Biol. 1989,5, 212.
Gouygou, J.P.; Martin, C.; Sinquin, C.; Durand, P.Oceanis 1989,15, 599.
Beyer, C.; Van den Ende, A.Clin. Chim. Acta 1980,129, 211.
Yamamoto, S.; Itano, H.; Kataoka, H.; Makita, M.J. Agric. Food. Chem. 1982,30, 435.
Draisci, R.; Cavalli, S.; Lucentini, L.; Stacchini, A.Chromatographia 1993,35, 584.
Akira, T.; Kentaro, A.; Tomofumi, S.; Kazuhiro, I.Anal. Chem. 2000,72, 732.
Nicole, J.M.; Zhili, H.; David, N.; James, P.L.Anal. Chem. 2000,72, 2765.
Anders, E.; Per, E.A.; Björm, J.; William, D.P.Anal. Chem. 1995,67, 3018.
Blau, K.; King, G.Handbook of Derivatives for Chromatography, Heyden, London,1977.
Knapp, D.R.Handbook of Analytical Derivatization Reactions, Wiley, New York,1979.
Vendrell, J.; Aviles, F.X.J. Chromatogr. 1986,358, 401.
Muramoto, K.; Kamiya, H.; Kawauchi, H.Anal. Biochem. 1984,141, 446.
Hirano, H.; Wittmann-Liebold, B.Biol. Chem. Hoppe-Seyler 1986,367, 1259.
Ishida, T.; Yamaguchi, M.; Iwata, T.; Nakamura, M.Anal. Chim. Acta 1989,223, 319.
Toyo’oka, T.; Suzuki, T.; Watanabe, T.; Liu, Y.M.Anal. Sci. 1996,12, 779.
Busto, O.; Mestres, M.; Guasch, J.; Borrull, F.Chromatographia 1995,40, 404.
Petridies, K.D.; Steinhart, H.Z. Lebensm. Unters. Forsch. 1995,201, 256.
Frattini, V.; Lionetti, C.J. Chromatogr. A 1998,809, 241.
Tateda, N.; Matsuhisa, K.; Hasebe, K.; Kitajima, N.; Miura, T.J. Chromatogr. B 1998,718, 235.
Lindroth, P.; Mopper, K.Anal. Chem. 1979,5, 1667.
Hill, D.W.; Walters, F.H.; Wilson, T.D.; Stuart, J.D.Anal. Chem. 1979,51, 1338.
Chem, R.F.; Scott, C.; Trepman, E.Biochim. Biophys. Acta 1979,576, 440–445.
Barth, H.G.; Barber, W.E.; Lochmuller, C.H.; Majors, R.E.; Regnier, F.E.Anal. Chem. 1988,60, 387.
Einarsson, S.; Josefsson, B.; Lagerkvist, S.J. Chromatogr. 1983,282, 609.
Einarsson, S.; Folestad, S.; Josefsson, B.; Lagerkvist, S.Anal. Chem. 1986,58, 1638.
Kirschbaum, J.; Busch, I.; Brückner, H.Chromatographia 1997,45, 263.
Inoue, H.; Kohashi, K.; Tsuruta, Y.Anal. Chim. Acta 1998,365, 219.
Kovacs, A.; Simon-Sarkadi, L.; Ganzler, K.J. Chromatogr. A,1999,836, 305.
Busto, O.; Guasch, J.; Borrull, F.J. Chromatogr. A 1996,737, 205.
Steven, A.; Dennis, P.M.; Michaud,Anal. Biochem. 1993,211, 279.
Romero, R.; Bagur, M.G.; Sanchez-Vinas, M.; Gazquez, D.Chromatographia 2000,51, 404.
Valle, M.; Malle, P.; Bouquelet, S.J. AOAC Int. 1997,80, 49.
Moret, S.; Bortolomeozzi, R.; Lercker, G.J. Chromatogr. 1992,591, 175.
Bockhardt, A.; Krause, I.; Klostermeyer, H.Z.Lebensm. Unters. Forsch. 1996,203, 65.
Xinjun Fan; Jinmao You; Jinwu Kang; Qingyu Ou; Qingcun ZhunAnal. Chim. Acta. 1998,367, 81.
Arthur, J.F.; Hans, V.; Hans, D.B.Anal. Chem. 1991,63, 292.
Ahnoff, M.; Grundevik, I.; Arfwidsson, A.; Fonselius, J.; Bengt, A.P.Anal. Chem. 1981,53, 485.
Ghosh, P.B.; Whitehouse, M.W.Biochem. J. 1968,108, 155.
Fardiaz, D.; Markakis, P.J. Food Sci. 1979,44, 1562.
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You, J., Zhang, Y. Determination of biogenic amines by RPHPLC with fluorescent detection after derivatization with 2-(9-carbazole)ethyl chloroformate (CEOC). Chromatographia 56, 43–50 (2002). https://doi.org/10.1007/BF02490245
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DOI: https://doi.org/10.1007/BF02490245