A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the determination of phenolic polycyclic aromatic hydrocarbons (OH-PAH) in urine of non-smokers and smokers |
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Authors: | Bernhard Ramsauer Katharina Sterz Heinz-Werner Hagedorn Johannes Engl Gerhard Scherer Mike McEwan Graham Errington Jim Shepperd Francis Cheung |
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Institution: | (1) Analytisch-Biologisches Forschungslabor GmbH, Goethestrasse 20, 80336 Munich, Germany;(2) British American Tobacco, Group Research and Development, Regents Park Road, Southampton, SO15 8TL, UK; |
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Abstract: | Polycyclic aromatic hydrocarbons (PAH) are products of the incomplete combustion of organic materials and, therefore, occur
ubiquitously in the environment and also in tobacco smoke. Since some PAH have been classified as carcinogens, it is important
to have access to suitable analytical methods for biomarkers of exposure to this class of compounds. Past experience has shown
that measuring a profile of PAH metabolites is more informative than metabolites of a single PAH. Assessment of environmental
and smoking-related exposure levels requires analytical methods with high sensitivity and specificity. In addition, these
methods should be fast enough to allow high throughput. With these pre-conditions in mind, we developed and validated a high-performance
liquid chromatographic method with tandem mass spectrometric detection (LC-MS/MS) for the determination of phenolic metabolites
of naphthalene, fluorene, phenanthrene and pyrene in urine of smokers and non-smokers. Sample work-up comprised enzymatic
hydrolysis of urinary conjugates and solid-phase extraction on C18 cartridges. The method showed good specificity, sensitivity,
and accuracy for the intended purpose and was also sufficiently rapid with a sample throughput of about 350 per week. Application
to urine samples of 100 smokers and 50 non-smokers showed significant differences between both groups for all measured PAH
metabolites, and strong correlations with markers of daily smoke exposure in smoker urine. Urinary levels were in good agreement
with previously reported data using different methodologies. In conclusion, the developed LC-MS/MS method is suitable for
the quantification of phenolic PAH metabolites of naphthalene, fluorene, phenanthrene, and pyrene in smoker and non-smoker
urine. |
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