Validation study of a HPLC method for biogenic amines quantification in bananas

Abstract

In this paper we describe an internal study for validating high performance liquid chromatography (HPLC) method for quantification of the biogenic amines in bananas. The evaluated features for validating the measuring method by means of the high performance liquid chromatography are as follows: linearity, precision, accuracy (repeatability and reproducibility), selectivity, sensitivity (detection limit, quantification limit), and robustness. The analyzed biogenic amines are: tryptamine, phenylethylamine, putrescine, cadaverine, histamine, serotonin, tyramine, spermidine and spermine. The calibration curves for the biogenic amines are linear and the values of the linearity coefficients (r2 ) are as follow: tryptamine r 2 = 0,9953, β-phenylethylamine r 2 = 0,9983, putrescin r2 = 0,9985, cadaverine r2 = 0,9985, histamine r2 = 0,9981, serotonin r2 = 0,9966, tyramine r2 = 0,9986, spermidine r2 = 0,9986, spermine r2 = 0,9982. The average recovery in the concentration levels 0.5 … 2 µg/ml for the banana samples recorded the following values: tryptamine 63-93%; βphenylethylamine 80-87%; putrescine 85-99%; cadaverine 96-110%; histamine 80-93%; serotonin 60-85%; tyramine 82-98%; spermidine 80- 105% and spermine 90-110%. The proposed method for biogenic amines quantification in bananas by HPLC is selective. The peaks for every biogenic amine are separated by the baseline and by the vicinity peaks. The resolution that characterize the selectivity is greater than 1. LOD (limit of detection) for every amine are as follows: tryptamine 0.006 µg/ml, β-phenylethylamine 0.050 µg/ml, putrescin 0.022 µg/ml, cadaverine 0.030 µg/ml, histamine 0.035 µg/ml, serotonin 0.015 µg/ml, tyramine 0.006 µg/ml, spermidine 0.005 µg/ml, spermine 0.009 µg/ml. LOQ (limit of quantitation) are as follows: tryptamine 0.012 µg/ml, β-phenylethylamine 0.100 µg/ml, putrescin 0.044 µg/ml, cadaverine 0.060 µg/ml, histamine 0.07 µg/ml, serotonin 0.030 µg/ml, tyramine 0.012 µg/ml, spermidine 0.01 µg/ml, spermine 0.018 µg/ml.