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  • Automated derivatization and fluorimetric determination of biogenic amines in milk by zone fluidics coupled to liquid chromatography.

Automated derivatization and fluorimetric determination of biogenic amines in milk by zone fluidics coupled to liquid chromatography.

Journal of chromatography. A (2014-07-06)
Maria Notou, Anastasia Zotou, Paraskevas D Tzanavaras, Demetrius G Themelis
ABSTRACT

A novel zone-fluidics/high pressure liquid chromatographic (ZF-HPLC) method is described for the simultaneous determination of six biogenic monoamines, in the presence of hexylamine as internal standard. Automated on-line derivatization of the analytes with naphthalene-2,3-dicarboxaldehyde/cyanide ions was performed using the ZF concept and the derivatives were injected on the HPLC column for separation/detection. The influence of the ZF operation conditions on the derivatization reaction was investigated. The isoindoles formed were separated on a Kromasil C18 column (250 mm × 4 mm i.d., 5 μm), using an isocratic mobile phase of methanol/water (80:20, v/v) at a flow rate of 1.0 mL min(-1). Monitoring and quantification was carried out by fluorescence detection at 424/494 nm. The limits of detection were at the pg level with a sample volume of 20 μL. The whole procedure was evaluated and fully validated for the determination of biogenic amines in milk samples.

MATERIALS
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