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

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.

Determination of phenolic compounds by ultra high liquid chromatography-tandem mass spectrometry: Applications in nuts

“…Validation of UHPLC-ESI-MS/MS method for analysis of selected fourteen phenolic compounds was achieved by investigating following parameters; linearity, limit of detection, limit of quantification, repeatability (intra-day and inter-day precision), and recovery (Li et al, 2005;Matuszewski, Constanzer, & Chavez-Eng, 2003;Ramsauer et al, 2011;Shou, Jiang, & Naidong, 2004;Wu, Yee, Jiang, & Cai, 2007). Fig.…”

Changes in levels of biomarkers of exposure and biological effect in a controlled study of smokers switched from conventional cigarettes to reduced-toxicant-prototype cigarettes

“…We measured 23 BoE and five BoBE in urine. The methods utilised for measurement of BoE for aromatic amines and polycyclic aromatic hydrocarbons have been reported elsewhere (Riedel et al, 2006;Ramsauer et al, 2011). Methods for BoE for nicotine and metabolites, tobacco-specific nitrosamines, 1,3-butadiene, crotonaldehyde, acrolein and acrylonitrile were adapted from published methods (Roethig et al, 2007;Kavvadias et al, 2009;Urban et al, 2003;Scherer et al, 2007;Ding et al, 2009).…”

Changes in Biomarkers of Exposure and Subjective Effects When Smokers Switch to Dual Use of Cigarettes and Either Snus or a Dissolvable Tobacco Product: A Summary of Three Clinical Studies

“…Concentrations of phenolic metabolites of polycyclic aromatic hydrocarbons (OH-PAHs) naphthalene, fluorene, phenanthrene, and pyrene were determined by GC-MS/MS according to RAMSAUER et al (32) with major modifications. Modifications included the following: the cited method describes an LC-MS/MS assay compared with the GC-MS/MS assay employed here; the volume of urine used was increased to 10 mL; 10 mL of n-hexane/acetonitrile (100:0.7) was used for elution from C18 solid phase extraction (SPE) cartridges, followed by derivatization with pentafluoropropionic acid anhydride; and all OH-PAHs were chromatographically separated.…”