CORESTA Meeting, Smoke/Technology, Budapest, 1993, ST 09

Determination of amino acids in tobacco by HPLC/fluorescence with precolumn derivatization

Philip Morris USA, Richmond, VA, USA
Precolumn derivatization has been of interest in amino acids determination for years, especially in the cases where complex matrix presents. In this study, an HPLC method is described for the analysis of amino acids in tobacco with a precolumn derivatization using a fluorogenic reagent, naphthalene-2,3-dicarboxaldehyde. Sample preparation consists of a single extraction at ambient temperature with the aid of sonication. Different solvents, aqueous and organic, have been evaluated for the extraction efficiency. An aliquot of tobacco extract is filtered and then mixed with the derivatizing reagents. The reaction is complete within 30 minutes under ambient conditions. High performance liquid chromatographic analysis of the derivatized amino acids is performed on a C-18 column followed by fluorescence detection (ex/em, 246/470 nm). This method has been used to analyze different types of tobacco materials (e.g. Burley, Bright and Oriental). Parameters and analytical results of this procedure will be discussed. Other derivatizing reagents for the quantitative analysis of amino acids in tobacco also are evaluated for comparison.