Summary
An HPLC method for the determination of primary amino acids in tobacco, based on precolumn derivatization with the fluorogenic reagent naphthalene-2,3-dicarboxaldehyde, is described. Sample preparation consists of a single step of extraction with waterethanol (1∶1, v/v) at ambient temperature (assisted by sonication), followed by filtration of an aliquot and addition of derivatization reagents. High performance liquid chromatographic separation of the derivatized amino acids is performed on a Novapak C-18 column using fluorescence detection with excitation and emission wavelengths set at 246 and 470 nm respectively. Extraction efficiency, matrix interferences and other important variables are studied. This method has been used to analyze different types of tobaccos including burley, bright and oriental with satisfactory results.
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Yang, S.S., Smetena, I. Determination of free amino acids in tobacco by HPLC with fluorescence detection and precolumn derivatization. Chromatographia 37, 593–598 (1993). https://doi.org/10.1007/BF02274107
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DOI: https://doi.org/10.1007/BF02274107