Validation of an ion-pair liquid chromatography-electrospray-tandem mass spectrometry method for the determination of heterocyclic aromatic amines in meat-based infant foods

Lisa Elviri (1)

1 Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica

We present a method based on ion-pair liquid chromatography electrospray tandem mass spectrometry (LC–MS/MS) for determining in meat-based infant foods heterocyclic aromatic amines encompassing quinoline (IQ, MeIQ), quinoxaline (MeIQx), pyridine (PhIP) and carboline derivatives (AαC, Harman, Norharman). d3-IQ, 13C2-MeIQx and d3-PhIP were used as labelled internal standards. The method uses extraction in acetone followed by a clean-up on an SCX solid-phase extraction column. LC separation was performed on a TSKgel ODS-80TS column (250 x 2.0 mm, 5Μm), the mobile phase being an ammonium formate-formic acid buffer (3.03 mM ammonium formate, pH=2.8) aqueous solution–acetonitrile gradient at a flow rate of 0.2 ml min−1. For unequivocal identification of each analyte, three ions were detected and chosen for selected reaction monitoring (SRM). Validation was carried out on lyophilized meat sample. Mean recoveries ranged between 78°4% to 98°2% for different analytes. Quantification limits generally lower than 8 ng/g were calculated in meat samples for the analytes investigated. The method exhibited good linearity and repeatability. Robustness test evidenced those factors influencing with a statistical significance chromatographic separation and response and suggested which parameters have to be strictly controlled for a reliable analysis of HAAs. In particular, mobile phase flow-rate was found to be statistically significant (Α = 0.05) for the capacity factor (k') of all analytes except for AΑC peak, whereas mobile phase pH resulted to be a critical parameter for the k' values of IQ, MeIQ and Norharman. The method was proved robust versus resolution between IQ and MeIQ peaks. Among mass spectrometric parameters, collision energy was found to significantly affect quantitative response of all analytes except that of IQ. The applicability of the method to the analysis of meat-based infant food samples was demonstrated.

Mots clés: Life Sciences

Lisa Elviri. Validation of an ion-pair liquid chromatography-electrospray-tandem mass spectrometry method for the determination of heterocyclic aromatic amines in meat-based infant foods. Food Additives and Contaminants, 2007, 24 (08), pp.833-841. ⟨10.1080/02652030701278339⟩. ⟨hal-00577287⟩