Technical guidelines on testing the migration of primary aromatic ...

Annex 1 – Primary aromatic aminesThis annex presents three HPLC-MS based methods and one HPLC-DAD methodfor the determination of primary aromatic amines (PAA) in the food simulants distilledwater and 3 % acetic acid.Table of contents1 PROTOCOL A BY LC-MS .......................................................................................................231.1 PRINCIPLE ...........................................................................................................................231.2 MATERIALS AND CHEMICALS ..............................................................................................231.2.1 Analytes and chemicals ..............................................................................................231.2.2 Solutions .......................................................................................................................241.2.3 Apparatus......................................................................................................................251.3 PROCEDURES......................................................................................................................251.3.1 Analysis.........................................................................................................................251.3.2 HPLC conditions ..........................................................................................................261.3.3 HPLC calibration..........................................................................................................261.3.4 Quantification of analyte level....................................................................................271.3.5 Confirmation of identity ...............................................................................................271.4 PRECISION DATA .................................................................................................................271.4.1 Validation ......................................................................................................................271.4.2 Detection limits.............................................................................................................271.4.3 Recovery.......................................................................................................................282 PROTOCOL B BY LC-MS .......................................................................................................292.1 PRINCIPLE ...........................................................................................................................292.2 MATERIALS AND CHEMICALS...............................................................................................302.2.1 Analytes and chemicals ..............................................................................................302.2.2 Solutions .......................................................................................................................302.2.3 Apparatus......................................................................................................................312.3 PROCEDURES......................................................................................................................312.3.1 Samples (3% acetic acid migration solutions).........................................................312.3.2 HPLC conditions ..........................................................................................................312.3.3 HPLC Calibration .........................................................................................................322.3.4 Quantification of the analyte level .............................................................................332.3.5 Confirmation of identity ...............................................................................................332.4 PRECISION OF THE DATA.....................................................................................................332.4.1 Validation ......................................................................................................................332.4.2 Detection limits.............................................................................................................343 PROTOCOL C BY LC-MS .......................................................................................................353.1 PRINCIPLE ...........................................................................................................................353.2 MATERIALS AND CHEMICALS ..............................................................................................353.2.1 Analytes and chemicals ..............................................................................................353.2.2 Solutions .......................................................................................................................363.2.3 Apparatus......................................................................................................................363.3 PROCEDURE........................................................................................................................373.3.1 Analysis of test samples.............................................................................................373.3.2 HPLC Conditions .........................................................................................................373.3.3 Quantification of the analyte level .............................................................................373.4 PRECISION OF THE DATA.....................................................................................................383.4.1 Limit of detection and quantification..........................................................................383.4.2 Bias and precision .......................................................................................................3921