Determination of biogenic amines in apples and wine with 8-phenyl-(4-oxy-acetic acid N-hydroxysuccinimide ester)-4, 4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene by high performance liquid chromatography Original Research Article

A precolumn-derivatization high performance liquid chromatographic method for biogenic amine analysis has been developed. Derivatization of biogenic amines was performed with a new fluorescent reagent, 8-phenyl-(4-oxy-acetic acid N-hydroxysuccinimide ester)-4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene (TMPAB-OSu), which was synthesized in our lab. The derivatization conditions and the influence of elution composition on the separation were investigated. In pH 8.5 H3BO3–Na2B4O7 media, the reaction of biogenic amines with TMPAB-OSu completed at 45 °C for 5 min. The labeled biogenic amines were separated on an ZORBAX Eclipse XDB-C8 column (150 mm × 4.6 mm, 5 μm) and detected with fluorescence at excitation and emission wavelengths of 497 and 509 nm, respectively. Detection limits of biogenic amines were 0.1–4 nmol L−1, at a signal-to-noise ratio of 3. The proposed method has been applied to the quantitative determination of spermine, phenethylamine, spermidine, cadaverine and putrescine in pericarp and pulp of mature and immature apples and wine with recoveries of 95–102% and R.S.D. of 0.7–3.9.