Abstract:

A method for detecting the content of biogenic amine in traditionally fermented Douchi by HPLC was developed.
The method was based on a reversed-phase column (Agilent XDB C18 column, 250 mm × 4.6 mm × 5 μm) for pre-column
derivatization with benzoyl chloride. Douchi samples were separated by HPLC using gradient elution (methanol and ultra
pure water), and detected with a UV-visible detection at 254 nm. The method allowed simultaneous determination of 8
amines in less than 25 min including putrescine, cadaverine, tryptamine, 2-phenyl-ethylamine, spermidine, spermine,
histamine and tyramine. A good linearity for each amine was observed between 1 μg/mL and 80 μg/mL. The detection
limits ranged from 0.03 to 0.10 μg/mL. The average recovery rate for biogenic amines in traditionally fermented Douchi
was 96.80%–106.10%. Due to its high sensitivity, precision and repeatability, the developed method was feasible for the
determination of biogenic amines in traditionally fermented Douchi. Our results indicated that biogenic amine types and
contents varied among different traditionally fermented Douchi. The variation of total biogenic amine in traditionally
fermented Douchi was 101.07–427.19 mg/kg.

Key words: traditionally fermented Douchi, biogenic amines, high performance liquid chromatography (HPLC)