Tobacco specific nitrosamines (TSNAs) are a group of carcinogens inherent to tobacco and tobacco products. There is strong evidence for TSNAs’ important role in inducing tobacco-related diseases. Of all TSNAs, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is the most carcinogenic. 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), the primary metabolite of NNK, is a cancer risk biomarker in addition to its well-established role as an exposure biomarker in epidemiologic studies. In our laboratory, a high throughput liquid chromatography tandem mass spectrometry (LC/MS/MS) method with robotic sample preparation and automatic data processing was validated for the measurement of four TSNAs in urine.  However, the TSNA method has limitations, including implementation challenges and cost. On the other hand, the properties of NNAL make it a better biomarker for studies of tobacco product exposure. Therefore we improved the existing method to a measurement of NNAL only by shortening the analysis time, and therefore, increasing the throughput. The sample size was decreased from previously reported 1.7 mL to 0.25 mL. The sample preparation time was shortened from previous reported 10 hours to 2 hours. The automatic data process by using Indigo Ascent™ combined peak integration and quality assurance, resulting in improved precision and reproducibility. Furthermore, the robotic sample preparation method increased throughput.  Our results suggest that this NNAL only method is suitable for application to large epidemiological investigations of health risks associated with the use of tobacco products.

Improved method for the measurement of tobacco-specific carcinogen biomarker urinary 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in tobacco users

2019 Fall National Meeting

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