Simultaneous Solid-Phase Extraction and Gas Chromatographic--Mass Spectrometric Determination of Hemoglobin Adducts from Tobacco-Specific Nitrosamines and Aromatic Amines

Abstract

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.

Haemoglobin adducts from aromatic amines in children in relation to area of residence and exposure to environmental tobacco smoke

“…In this study internal exposure to aromatic amines has been studied in children by an established analytical method (Tannenbaum and Skipper 1994) slightly modi®ed by us (Kutzer et al 1997), and acknowledged after further minor modi®cations by the German Commission for the Investigation of Health Hazards of Chemical Compounds in the Work Area (Lewalter and Gries (2000). Adducts from 4-ABP, o-, m-and p-toluidine isomers, as well as from o-anisidine, were detected in all samples.…”

“…Aromatic amine Hb adducts were determined as previously described, with some minor modi®cations (Kutzer et al 1997). Brie¯y, Hb solutions obtained after centrifugation of lysed red Table 1 Sociodemographic data of the study population.…”

4-Hydroxy-1-(3-pyridyl)-1-butanone, an Indicator for 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone–Induced DNA Damage, Is Not Detected in Human Pancreatic Tissue

“…We have analyzed 58 samples of pancreatic DNA by gas chromatography-mass spectroscopy (7). As a positive control, we used DNA isolated from mouse lung treated by a single i.p.…”

Analytical Aspects of Aromatic Amines

“…A similar procedure was applied for isolation of 7a from the 7a-Hb adduct in smokers' blood after spiking the Hb lysate with 4 -fluoro-4-biphenylamine as internal standard; the pentaphenylpropionamide derivatives of the analyte and standard (Section III.B.1) were subjected to end analysis by GC-MS 35 . The lysate of PAA and tobacco-specific nitrosamines recovered by hydrolysis of their Hb adducts in smokers' blood was subjected to SPE on C 18 , using stainless steel vessels instead of the conventional plastic material, which may dissolve part of the analytes 119 .…”

“…5a), taking place at 160 to 180 • C. The following NIR spectral ranges were useful for this study: primary amine first overtones (ν N−H ) at 7000 to 6400 cm −1 , double bond first overtone (ν C=C−H ) at 6100 cm −1 , aromatic first overtones (ν C−H ) at 6000 to 5750 cm −1 , aliphatic first overtones (ν C−H ) at 5750 to 5350 cm −1 and primary aromatic amine combination bands first overtones (ν N−H + δ NH2 ) at 5150 to 4800 cm −1 . The process consisted mainly of a second-order Michael addition, as depicted in equation 14, and not the plausible imide opening to yield a maleic dianilide (119), as shown in equation 15. A Michael addition between maleimide moieties and secondary amine moieties present in the products (118) also takes place, however at a rate of about one fourth of that of the primary amine moieties.…”