A simple, economical and reproducible benzoylation procedure was developed for the derivatization of biogenic amines (BAs) prior to high-performance liquid chromatography determination. The significant factors affecting biogenic amine benzoylation yield were optimized by central composite design. The derivatized BAs were extracted from basic aqueous solutions by 2-undecanol for direct HPLC injection. The obtained optimal conditions significantly decreased reagent consumption, uncertainty and time of analysis. The optimized method was used for further determination of BAs in real samples of non-alcoholic beers. The method showed good linearity (correlation coefficients > 0.997) and good recoveries (from 87.3 to 96.3%). The repeatability and reproducibility of the method were >4.6% and >6.7% respectively. Moreover, the detection limits of BAs were calculated between 0.02 and 0.09 μg ml−1 in non-alcoholic beers samples.