UPLC-MS/MS for high-throughput analysis of aromatic amines in cigarette smoke
Aromatic Amines (AAs) are included in the “Established List of the Chemicals and Chemical Compounds Identified by FDA as Harmful and Potentially Harmful Constituents [HPHCs] in Tobacco Products and Tobacco Smoke” (Federal Register (Vol. 77, No. 64) Docket No. FDA-2012-N-0143). To date, no standardized method for AA determination in mainstream cigarette smoke has been developed. Previously reported techniques for the quantitative analysis of trace amounts of AAs in cigarette smoke (ng per cigarette) include gas chromatography with mass spectrometry (GC-MS) involving multistep solid phase extractions (SPE) or high performance liquid chromatography with multistage mass spectrometry (HPLC-MS/MS) with liquid-liquid extraction. The purpose of this work was to evaluate a higher throughput approach using ultra-performance liquid chromatography (UPLC) with MS/MS with a single step SPE clean-up. This method demonstrated applicability to all AAs on the FDA HPHC list and was validated for the 3 AAs on the current abbreviated HPHC list. All requirements for method validation were met such as linearity, accuracy, precision, limits of detection (LOD), and limits of quantitation (LOQ). For example, the linearity was demonstrated with a coefficient of determination of greater than 0.990 for the calibration ranges of 1.5 to 150 ng/cigarette for 1-aminonaphthalene, 0.75 to 75 ng/cigarette for 2-aminonaphthalene, and 0.6 to 30 ng/cigarette for 4-aminobiphenyl under the ISO smoking regime.