title: Validation of an ion-pair liquid chromatography-electrospray-tandem mass spectrometry method for the determination of heterocyclic aromatic amines in meat-based infant foods.
abstract: A method based on ion-pair liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) is reported for determining heterocyclic aromatic amines (HAAs) in meat-based infant foods. The HAAs encompassed quinoline (IQ, MeIQ), quinoxaline (MeIQx), pyridine (PhIP), and carboline derivatives (AalphaC, Harman, Norharman) with d(3)-IQ, (13)C(2)-MeIQx, and d(3)-PhIP used as labelled internal standards. The method used extraction into 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, 5 microm), 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 samples. Mean recoveries ranged between 78 +/- 4% and 98 +/- 2% for different analytes. Limits of quantification generally lower than 8 ng g(-1) were demonstrated in meat samples for the analytes investigated. The method exhibited a good linearity and repeatability. Robustness testing identified those factors which were statistically significant in influencing chromatographic separation and response, and indicated which parameters have to be strictly controlled for a reliable analysis of HAAs. In particular, the mobile-phase flow rate was found to be statistically significant (alpha = 0.05) for the capacity factor (k') of all analytes except for AalphaC peak, whereas the mobile-phase pH resulted to be a critical parameter for the k' values of IQ, MeIQ, and Norharman. The method was proved to be robust vs. 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.
authors.affiliation: Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica, Università degli Studi di Parma, 43100 Parma, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Calbiani
authors.forename: M
authors.initials: M
authors.lastname: Careri
authors.forename: L
authors.initials: L
authors.lastname: Elviri
authors.forename: A
authors.initials: A
authors.lastname: Mangia
authors.forename: I
authors.initials: I
authors.lastname: Zagnoni
mesh_terms: D000588:Amines / Q000032:analysis*; D000818:Animals; D002273:Carcinogens / Q000032:analysis*; D002853:Chromatography, Liquid / Q000379:methods; D006571:Heterocyclic Compounds / Q000032:analysis*; D006801:Humans; D007223:Infant; D007225:Infant Food / Q000032:analysis*; D008461:Meat Products / Q000032:analysis*; D009153:Mutagens / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D000588:Amines; D002273:Carcinogens; D006571:Heterocyclic Compounds; D009153:Mutagens