A RT-HPLC Method for the Determination of Biogenic Amine Produced by Lactic Acid Bacteria

Abstract:

Biogenic amines are produced by some lactic acid bacteria of decarboxylase activities which are potential risk factors for lactic acid bacteria-fermented foods. In this paper, the biogenic amine-producing capacities of 61 lactic acid bacteria strains were analyzed on a decarboxylase medium. Then the methodology of RT-HPLC determination for biogenic amines in fermented broth and milk was studied. The biogenic amines in test samples were extracted by 10% trichloroacetic acid and then derived with benzoyl chloride. The HPLC analysis for the samples were conducted as the following condition: mobile phase was a gradient elution program of methanol and water, the flow rate was 0.8 mL/min. UV detector was used in the final de-termination on 254 nm. The results showed that biogenic amines were separated well on such condition. The method was in a linear way for the amines studied at given concentrations (R2 > 0.995). The average recov-ery ratios ranged from 97.92%?101.14% for all samples. The RSDs were less than 5%. It can be seen that the RT-HPLC method was rapid, stable, high sensitive. Combination of this HPLC method and microbial primary screening on a decarboxylase medium could evaluate the biogenic amine-producing capacity of lac-tic acid bacteria more accurately.

Key words:Lactic acid bacteria, Biogenic amine, Decarboxylase, RT-HPLC