Simultaneous solid-phase extraction and gas chromatographic-mass spectrometric determination of hemoglobin adducts from tobacco-specific nitrosamines and aromatic amines.

A new method for the simultaneous determination of hemoglobin adducts from aromatic amines and tobacco-specific nitrosamines (TSNA) is described. After mild base-catalysed hydrolysis releasing aromatic amines and the TSNA adduct 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB), the extraction, cleanup, and concentration are performed by a one-step procedure using C18 cartridges. Determination in the picograms per gram of hemoglobin range by gas chromatography-mass spectrometry with negative chemical ionization requires a separate derivatization procedure with pentafluoropropionic anhydride and pentafluorobenzoylchloride for aromatic amines and HPB, respectively. The method is shown to be quantitative, reproducible, and applicable to the determination of hemoglobin adducts from monocyclic and bicyclic aromatic amines as well as TSNA in smokers and nonsmokers.