Journal of the Korean Society of Tobacco Science (한국연초학회지)

The Korean Society of Tobacco Science (한국연초학회)
권호 표지

The Analysis of Free Amino Acids in Tobacco

잎담배 중 유리 아미노산 분석

  • Received : 2010.11.09
  • Accepted : 2010.11.29
  • Published : 2010.12.24
Download PDF
⟨ Previous Next ⟩

Abstract

A simple and sensitive automatic amino acid analyzer method for the determination of free amino acids in tobacco was described. Sample preparation consisted of a single step of extraction with 0.1 mol HCl at ambient temperature in 60 min by sonication, followed by filtration of an aliquot. Automated amino acid analyzer was used to construct a post-column ninhydrin reaction unit to monitor amino acids separated by liquid chromatography using a series of eluting buffers. By optimization of sample preparation, separation of 19 amino acids was achieved. Limits of quantitation was 0.01 mg/g, coefficients of variation ranged from 0.5 % to 8.9 % and recoveries range from 85 % to 106 %. The method was applied to the analysis of amino acids contents of tobacco leaves in different varieties.

Keywords

References

  1. Alexander, R.R. (1985) Griffiths, J.M., Wilkinson, M.L. Identification of amino acids by thin-layer chromatography, Basic Biochemical Methods, Wiley, New York, pp. 24-28.
  2. Bailey, J.L. (1962) Estimation of amino acids by ninhydrin, techniques in protein chemistry, Elsevier, Amsterdam, pp. 73-81.
  3. Cohen, S.A., Bidlingmeyer, B.A., and Tarvin, T.L. (1986) PITC derivatives in amino acid analysis. Nature 320: 769-770. https://doi.org/10.1038/320769a0
  4. Davis, D.D. and Nielsein M.T. (1999) Tobacco production, chemistry and technology, Blackwell Science, Oxford: U. K., p 273.
  5. Dejong, C., Hughes, G.J., Wieringen, E. and Wilson, K.J. (1982) Amino acid analyses by high-performance liquid chromatography-an evaluation of the usefulness of precolumn DNS dervatisation. J Chromatogr 241: 345-359. https://doi.org/10.1016/S0021-9673(00)81759-8
  6. Hirs, C.H.W. Detection of peptides by chemical methods, Methods in Enzymology, Enzyme Structure vol. XI, Academic Press, San Diego, pp. 325-329, 1967.
  7. Hurst, P.L., Sinclair, B.K. and Eason, J.R. (1995) Amino acids interfere with the ninhydrin assay for asparagines, Food Chemistry 53: 467-469. https://doi.org/10.1016/0308-8146(95)99843-O
  8. Laseter, J.L., Weete, J.D., Albert, A. and Walkinshaw, C.H. (1971) Gas chromatographic determination of trimethylsilyl derivatives of free amino acids from a botanical source, Analytical Letter 4: 671-681. https://doi.org/10.1080/00032717108058665
  9. Moore, S. and Stein, W.H. (1948) Photometric ninhydrin method for use in the chromatography of amino acids, J. Biological Chem. 176: 367-388.
  10. Noma, M., Noguchi, M. and Tamaki, E. (1973) Isolation and characterization of D-alanyl- D-alanine from tobacco leaves, Agr. Biol. Chem. 37: 2439. https://doi.org/10.1271/bbb1961.37.2439
  11. Panasiuk, R., marowicz, R., Kostyra, H. and Sijtsma, L. (1998) Determination of α-amino nitrogen in pea protein hydrolysates: a comparison of three analytical methods, Food Chemistry 62: 36337.
  12. Roth, M. (1971) Fluorescence reaction for amino acids, Anal. Chem. 43: 880-882. https://doi.org/10.1021/ac60302a020
  13. Rubinstein, M., Chen-Kiang, S., Stein, S. and Udenfriend, S. (1979) Characterization of proteins and peptides by high-performance liquid chromatography and fluorescence monitoring of their tryptic digests, Anal. Biochem. 95: 117-121. https://doi.org/10.1016/0003-2697(79)90193-3
  14. Tso, T.C. (1991) Production, physiology, and biochemistry of tobacco plant, Ideals Inc., Beltsville, p 497.
  15. Watanabe, Y. and Imai, K. (1981) Highperformance liquid chromatography and sensitive detection of amino acids derivatized with 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole, Anal. Bioehem. 116:471-472. https://doi.org/10.1016/0003-2697(81)90390-0
  16. Yang, S.S. and Smetena, I. Determination of free amino acids in tobacco by HPLC with fluorescence detection and precolumn derivatization. Chromatographia, 37: 593-98.
  17. Zhu, X.L., Zhu, Q.S. and Gao Y. (2008) Fast analysis of free amino acids in tobacco by HPLC with fluorescence detection and automated derivatization, J. the Chinese Chem. Society, 55: 1094-1100. https://doi.org/10.1002/jccs.200800160