Fast Analysis of Free Amino Acids in Tobacco by HPLC with Fluorescence Detection and Automated Derivatization |
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| Authors: | Xiao‐Lan Zhu Qing‐Sheng Zhu Yun Gao |
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| Affiliation: | 1. Research Center of Tobacco and Health, University of Science and Technology of China, Hefei 230052, P. R. China;2. School of Management, Hefei University of Technology, Hefei 230009, P. R. China;3. Technique Center of Modern Education, Anhui Institute of Architecture & Industry, Hefei 230022, P. R. China |
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| Abstract: | A fast, simple, and sensitive HPLC method for the determination of free amino acids in tobacco was described. A fully automated sample processor performed precolumn derivatization of both primary and secondary amino acids with o‐phthalaldehyde/3‐mercaptopropionic acid and 9‐fluorenylmethyl chloroformate (FMOC‐Cl), respectively. All reactions were fully automated by means of an injector programme and accomplished in 10 min. Sample preparation consisted of a single step of extraction with 0.1 mol/L HCl at ambient temperature (assisted by sonication) in 30 min, followed by filtration of an aliquot and derivatization. By optimization of sample preparation and HPLC conditions, separation of 20 amino acids in 30 min was achieved. Detection limits ranged from 0.50 to 1.40 μg/g; coefficients of variation ranged from 1.8% to 3.9%; recoveries ranged from 84.6% to 108.5%. The method was applied to the analysis of amino acids contents of tobacco leaves in different varieties and flue‐curing period. |
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| Keywords: | Amino acids Tobacco Column liquid chromatography Fluorescence detection Pre‐column derivatization |
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