Abstract
Background: Since the urinary concentration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is a reliable biomarker of exposure to tobacco smoke, we developed a relatively simple high-throughput chromatographic method to quantify total urinary NNAL concentrations in the general population.
Methods: The high-throughput analytical method was developed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) to identify and quantify total urinary NNAL concentrations in 10 non-smokers and 15 otherwise healthy smokers.
Results: Loss of nitric oxide at m/z 30 was found to be the predominant mass transitioned, and therefore was used as the SIM transition to quantify both NNAL and NNAL-methyl-d3 in urine. The analytical method did not require sample derivatization. Standard curves for total NNAL concentrations were linear between 20 and 1500 pg/mL, with coefficients of determination >0.95. Precision and accuracy ranged from 2.2% to 8.6% (CV) and from –5.6% to 10.9% (percent error), respectively. The lowest limit of quantification was 6.7 pg/mL, and 2.0 pg/mL the lowest limit of detection (LLOD). Total urinary NNAL concentrations in non-smoker subjects were <LLOD, whereas in smokers varied between <LLOD to 112.1 pg/mL.
Conclusions: An UPLC-MS/MS analytical method to quantify total urinary NNAL concentrations in smokers that does not require sample derivatization is presented herein. The method could be useful in clarifying the toxicities associated with human exposure to cigarette smoking. However, quantification might be adversely affected by co-eluting interfering compounds or selective ion suppression or enhancement as a result of having only one ion transition to monitor NNAL and NNAL-methyl-d3 in urine.
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