Determination of Biogenic Amines in Digesta by High Performance Liquid Chromatography with Precolumn Dansylation

Abstract A rapid high-performance liquid chromatographic method for the determination of nine biogenic amines in digesta sample from pig cecum and colon was developed using a simple direct derivatization with dansyl chloride. After precipitation with trichloroacetic acid and extraction by n-hexane, the amines were separated on a C18 column using a water–acetonitrile gradient elution program. The calibration curve for each amine was linear over the range of 50 to 250 µmol/L, and the relative standard deviation for intra-assay precision was below 0.5%. The recovery ranged from 86.79% to 117.62% while the limit of detection was 0.1 µmol/L for the amines except for spermine with a value of 0.03 µmol/L. This procedure was successfully applied to identify and quantify biogenic amines in the hindgut digesta of the pig. Keywords: Biogenic aminesDigestaHigh performance liquid chromatography ACKNOWLEDGMENT The author thanks masters' student Fan Zhao for assistance with the Waters data processing system (Empower 3), and the Key Laboratory of Meat Processing and Quality Control for its kind offer of Waters alliance HPLC system. Notes a Concentration of the nine standard amine solution using were 50, 100, 150, 200, and 250 µmol/L. b Square of regression coefficient. c Limit of detection, three times the noise level, µmol/L. d Limit of quantification, ten times the noise level, µmol/L. a Two levels of standard solution (0.1 and 0.2 mmol/L) were added. b Means of six repetitions. c SD, standard deviation. d RSD, relative standard deviations. a One sample of pig cecum digesta spiked with the lowest amine standard solution was used for testing repeatability and reproducibility during the same day and over a period of 5 days (50 µmol/L). b Means of six repetitions, µmol/L. c SD, standard deviation. d RSD, coefficient of variation. a Results were mean values from six individuals, values are µmol per g digesta. b Not detected (<0.01 µmol/g). c Standard Error of the Mean.