Urinary biomarkers to assess exposure of cats to environmental tobacco smoke.

OBJECTIVE: To evaluate the use of urinary biomarkers to assess exposure of cats to environmental tobacco smoke (ETS). ANIMALS: 61 healthy client-owned cats (19 from households in which smoking was reported and 42 from households in which there was no smoking). PROCEDURES: Urine samples were obtained from each cat and assayed for total nicotine (nicotine plus nicotine glucuronide) and total cotinine (cotinine plus cotinine glucuronide) content by use of gas chromatography-mass spectrometry. In addition, total urinary content of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a major metabolite of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, was measured by use of gas chromatography with nitrosamine-selective detection. RESULTS: Cats from households in which smoking was reported had significantly higher concentrations of total nicotine (70.4 ng/mL), total cotinine (8.53 ng/mL), and total NNAL (0.0562 pmol/mL) in urine, compared with concentrations for cats that lived in households in which there was no smoking (4.89 ng/mL, 0.74 ng/mL, and 0.0182 pmol/mL, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of these data provided biochemical evidence of exposure to ETS and uptake of tobacco-specific carcinogens by cats that live in households with smokers. Biomarkers could facilitate investigation of the health effects of ETS in cats and other species.

4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronides in the urine of infants exposed to environmental tobacco smoke.

Biomarkers of carcinogen uptake could provide important information pertinent to the question of exposure to environmental tobacco smoke (ETS) in childhood and cancer development later in life. Previous studies have focused on exposures before birth and during childhood, but carcinogen uptake from ETS in infants has not been reported. Exposures in infants could be higher than in children or adults because of their proximity to parents who smoke. Therefore, we quantified 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronides (total NNAL) in the urine of 144 infants, ages 3 to 12 months, who lived in homes with parents who smoked. Total NNAL is an accepted biomarker of uptake of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. Cotinine and its glucuronide (total cotinine) and nicotine and its glucuronide (total nicotine) were also quantified. Total NNAL was detectable in 67 of 144 infants (46.5%). Mean levels of total NNAL in the 144 infants were 0.083 +/- 0.200 pmol/mL, whereas those of total cotinine and total nicotine were 0.133 +/- 0.190 and 0.069 +/- 0.102 nmol/mL, respectively. The number of cigarettes smoked per week in the home or car by any family member when the infant was present was significantly higher (P < 0.0001) when NNAL was detected than when it was not (76.0 +/- 88.1 versus 27.1 +/- 38.2). The mean level of NNAL detected in the urine of these infants was higher than in most other field studies of ETS exposure. The results of this study show substantial uptake of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in infants exposed to ETS and support the concept that persistent ETS exposure in childhood could be related to cancer later in life.

Improved method for determination of 1-hydroxypyrene in human urine.

We have developed an improved method for the analysis of human urine for 1-hydroxypyrene (1-HOP), an accepted biomarker of polycyclic aromatic hydrocarbon uptake. This method takes advantage of commercially available 96-well format devices, which expedite sample preparation before quantitation by HPLC with fluorescence detection. In addition to improved speed of analysis, which is critical for the application of this assay in molecular epidemiology studies, the method described here uses an internal standard, 1-hydroxybenz[a]anthracene, improved sample preparation methods, and optimized HPLC and fluorescence detection conditions. The resulting method for analysis of 1-HOP is sensitive (detection limit, 0.05 pmol/mL urine), accurate (as determined by known addition of 1-HOP to urine), and precise [relative SD (RSD), 4.13%]. A longitudinal study of 1-HOP levels in the urine of 10 nonsmokers showed considerable day-to-day (mean RSD, 55.1 %) and week-to-week (mean RSD, 38.2 %) intra-individual variation, indicating the necessity for multiple sampling in studies concerned with relatively small differences in polycyclic aromatic hydrocarbon exposure.

Effects of reduced cigarette smoking on levels of 1-hydroxypyrene in urine.

We investigated the effects of smoking fewer cigarettes/day (CPD) on urinary levels of 1-hydroxypyrene (1-HOP), a biomarker of carcinogenic polycyclic aromatic hydrocarbon (PAH) uptake. We randomly assigned 151 smokers to either a reduction group or a waitlist group. In the reduction group, we measured urinary 1-HOP at two baseline intervals. Then, the smokers were expected to reduce their CPD by 25% in weeks 0-2, 50% in weeks 2-4, and 75% in weeks 4-6 and to maintain reduced smoking through week 26. In the waitlist group, four baseline measurements were taken and then the smokers joined the reduction group. Urinary 1-HOP was quantified at weeks 4, 6, 8, 12, and 26 after baseline. Statistically significant reductions in urinary 1-HOP were observed at most time points examined in groups of smokers who reduced to different extents. Reductions in the waitlist group were also generally significantly greater than baseline levels. The reductions in 1-HOP were usually modest (ranging from 14% to 35% in all groups and time points examined), which partially reflects the fact that there are sources of pyrene exposure other than cigarette smoke. Thus, cessation of smoking would only be expected to result in partial reductions of 1-HOP. The observed reductions in 1-HOP were not fully consistent with reductions in CPD probably due to uncontrolled dietary factors. Collectively, the results demonstrate that some smokers can achieve substantial reductions in 1-HOP, reflecting reduced uptake of polycyclic aromatic hydrocarbons, by reducing CPD, but there was not a consistent relationship between these parameters.

Quantitation of metabolites of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone after cessation of smokeless tobacco use.

Two major metabolites of the tobacco-specific lung carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) were previously shown to be highly persistent in human urine after cessation of cigarette smoking. We hypothesized that NNK or its metabolite, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), was sequestered in the lung. In this study, we further evaluated this hypothesis by quantifying the NNK metabolites, NNAL and its glucuronides (NNAL-Gluc), in urine and plasma after cessation of smokeless tobacco use, in which NNK is administered p.o. rather than by inhalation. Thirteen male nonsmokers, 11 snuff dippers and 2 tobacco chewers, participated in the study. Urine and plasma were obtained at baseline and at intervals 2-126 days after cessation of smokeless tobacco use. The distribution half-lives t(1/2alpha) (days) of NNAL (1.32 +/- 0.85 versus 3.35 +/- 1.86) and NNAL-Gluc (1.53 +/- 1.22 versus 3.89 +/- 2.43) were significantly shorter in smokeless tobacco users than in smokers. There were no significant differences in the terminal half-lives t(1/2beta) (days) of NNAL (26.3 +/- 16.7 versus 45.2 +/- 26.9) and NNAL-Gluc (26.1 +/- 15.1 versus 39.6 +/- 26.0) in smokeless tobacco users and smokers. Baseline levels as well as renal clearance of the NNK metabolites correlated with number of tins or pouches of smokeless tobacco consumed. Ratios of (S)-NNAL:(R)-NNAL and (S)-NNAL-Gluc:(R)-NNAL-Gluc in urine were significantly (3.1-5.7 times) higher 7 days after cessation than at baseline in both smokeless tobacco users and smokers, indicating stereoselective retention of (S)-NNAL. Collectively, the results of this study suggest that there is a receptor in the human body, possibly in the lung, for (S)-NNAL, the more carcinogenic NNAL enantiomer. These data may have considerable implications for understanding mechanisms of tumor induction by NNK.