Determination of Tobacco Specific Hemoglobin Adducts in Smoking Mothers and New Born Babies by Mass Spectrometry

Biological markers for assessment of exposure to a variety of environmental carcinogens has been widely applied in both basic as well as clinical research. Exposure to tobacco smoke presents an ideal environment with which to develop, characterize, and refine biological markers, especially of those carcinogens found in tobacco. In the present study, a sensitive gas chromatography/mass spectrometry (GC/MS) method was developed to measure nitrosamine- hemoglobin adducts (HPB-Hb (4-Hydroxy-3-pyridinyl-1-butanone) at trace levels in red blood cells of both African-American and Caucasian smoking and nonsmoking mothers and their infants. Gas chromatographic and mass spectrometric methods with chemical ionization (CI) of methane reagent gas in both positive and negative ion mode as well as electron ionization (EI) were studied to determine differences in sensitivity of detection among the various ionization methods. Detection limits using both positive and negative chemical ionization modes were found to be 30 femtomoles of HPB, whereas detection using electron impact modes yielded a detection limit of 80 femtomoles of HBP. Comparative derivatization of HPB was performed using O-bis(Trimethylsilyl)-trifluoroacetamide (BSTFA) and 2, 3, 4, 5, 6-Pentafluorobenzoylchloride (PFBC). Both Negative CI and Positive CI modes of analysis were compared to the more widely accepted EI modes of mass spectrometric analysis.