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

• Published in: Journal of Chromatography A Vol. 1356; pp. 272 - 276
• Main Authors: Notou, Maria, Zotou, Anastasia, Tzanavaras, Paraskevas D., Themelis, Demetrius G.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 22.08.2014; Elsevier
• Subjects: Animals; Autoanalysis; Biogenic amines; Biogenic Amines - analysis; Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Chromatography, High Pressure Liquid - standards; Fluorometry; Food industries; Fundamental and applied biological sciences. Psychology; Liquid chromatography; Milk; Milk - chemistry; Milk and cheese industries. Ice creams; Naphthalene-2,3-dicarboxaldehyde; Naphthalenes - chemistry; Reference Standards; Sequential injection analysis; Naphthalene-2,3-dicarboxaldehyde; Sequential injection analysis; Liquid chromatography; Biogenic amines; Milk; Validation; Automation; Fluorescence detector; Chemical analysis; HPLC chromatography; Derivatization; Trademark; Fluorescent labelling; Biogenic amine; Simultaneous measurement; Sequential injection; Quantitative analysis
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2014.06.047

•A novel zone-fluidics (ZF)–HPLC method is described for the determination of biogenic amines.•On-line derivatization of the amines with naphthalene-2,3-dicarboxaldehyde (NDA) is performed using ZF.•Using ZF the consumption of the expensive NDA is minimized to a few micro-liters per run. 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 (250mm×4mm i.d., 5μm), using an isocratic mobile phase of methanol/water (80:20, v/v) at a flow rate of 1.0mLmin−1. Monitoring and quantification was carried out by fluorescence detection at 424/494nm. The limits of detection were at the pg level with a sample volume of 20μL. The whole procedure was evaluated and fully validated for the determination of biogenic amines in milk samples.