Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1025, Issue 2, Pages 201-208Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2003.10.094
Keywords
derivatization; LC; amino acids; biogenic amines; Polyamines; N-(9-fluorenylmethoxycarbonyloxy)succinimide
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A high-performance liquid chromatography method for the simultaneous analysis of amino acids and biogenic polyamines, using a new procedure for pre-column derivatization of amino groups with N-(9-fluorenylmethoxycarbonyloxy)succinimide is described. The separation of 20 amino acids and 4 biogenic polyamines was achieved within 32 min on a sequence of three short (50 mm) reversed-phase C-18, 5 mum columns by elution buffers based on dibutylamine phosphate. The method linearity, calculated for each amino acid and polyamine, has a correlation coefficient higher than 0.991, in concentrations ranging from 0.2 to 50 muM, except for spermine and methionine, where the correlation coefficients were r = 0.984 and r = 0.979, respectively. The stability of derivatives in acidified samples at 4degreesC and room temperature was demonstrated. The limit of quantitation was estimated to be around 50 pM in 50 mul sample injection. The repeatability of the method, expressed as R.S.D., ranged from 1.1 to 6.7%. The presented method was applied for the quantitation of amino acid and polyamine contents in beer, wine, and cell culture samples, using 2-aminoheptanoic acid or 1,7-diaminoheptane as internal standard. (C) 2003 Elsevier B.V. All rights reserved.
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