Analysis for Eight Biogenic Amines in Traditional Pixian Broadbean by a Pre-column Derivatization-HPLC Method Based on Response Surface Optimization

A potential risk of biogenic amine contamination exists in Pixian broadbean due to its the traditional fermentation process. The concentration, time and temperature of dansyl Chloride (Dns-Cl) derivation, as well as reaction system pH and extraction solvent were optimized by response surface methodology in order to establish the pre-column derivatization reversed-phase high performance liquid chromatography (RP-HPLC) for biogenic Amines (BAs) analysis. Under these optimum conditions, eight kinds of biogenic amines, including histamine, tyramine, cadaverine, putrescine, phenethylamine, tryptamine, spermine and spermidine in Pixian broadbean samples with three different fermentation times were analyzed. The results show that the optimum conditions are: Dns-Cl concentration is at 11 mg/mL, and derivatization conditions are 38 min at 41 °C, and pH is 9. Under these conditions, each biogenic amine in the Pixian broadbean could be detected in a good linear correlation covering the concentration range of 1 ~ 500 mg/L, (R² ≥0.997 7). The detection limits ranged from 0.1 to 0.6 mg/L, and the limit of quantification ranged from 0.4 to 2.0 mg/L. The recovery rates were 90.44% to 109.29%, with relative standard deviations (RSD) ranging from 1.02% to 2.54%. In conclusion, the developed method owns high sensitivity, good reproducibility and accurate and reliable results.It can be used for the detection of biogenic amines in Pixian broadbean.