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Published on: March 20, 2018
An LC-MS/MS Method for the Quantification of Tobacco-Specific Carcinogen Protein Adducts
Breanne Freeman1, Chengguo Xing1
1Department of Medicinal Chemistry, Center for Natural Products, Drug Discovery and Development (CNPD3), College of Pharmacy, University of Florida, Gainesville, Florida 32610, United States.
Abstract:
4-(Methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and its major metabolite 4-(methylnitrosamino)-l-(3-pyridine)-l-butanol (NNAL) are tobacco-specific lung carcinogens. Methods have been developed to quantify NNK- and NNAL-specific DNA adducts in preclinical samples but are not feasible to translation due to limited access to target tissues for sufficient DNA. In addition, NNAL-specific DNA or protein adducts have never been detected in clinical samples, which are critical to assess the physiological relevance of NNAL bioactivation and carcinogenesis. We herein reported a highly sensitive and specific LC-MS/MS method to quantify the hydrolyzed product, 1-(3-pyridyl)-1,4-butanediol (PBD), from NNAL-induced protein adduct. This method was applied to a variety of biological samples to assess tobacco exposure and NNAL bioactivation.
Insights
Tobacco-specific lung carcinogens 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and its metabolite 4-(methylnitrosamino)-l-(3-pyridine)-l-butanol (NNAL) pose risks. A new LC-MS/MS method detects NNAL-induced protein adducts in clinical samples, aiding cancer research.
Area of Science:
- Environmental Health
- Toxicology
- Analytical Chemistry
Background:
- 4-(Methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and 4-(methylnitrosamino)-l-(3-pyridine)-l-butanol (NNAL) are key tobacco-specific lung carcinogens.
- Existing methods for quantifying NNK/NNAL DNA adducts are not clinically feasible due to limited sample access.
- NNAL protein adducts, crucial for understanding carcinogenesis, have not been detected in clinical samples.
Purpose of the Study:
- To develop a sensitive and specific method for detecting NNAL-induced protein adducts in clinical samples.
- To enable assessment of tobacco exposure and NNAL bioactivation in human populations.
- To bridge the gap between preclinical findings and clinical relevance of NNAL carcinogenesis.
Main Methods:
- Development of a highly sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method.
- Quantification of 1-(3-pyridyl)-1,4-butanediol (PBD), a hydrolyzed product of NNAL-induced protein adducts.
- Application of the method to various biological samples for biomarker analysis.
Main Results:
- A novel LC-MS/MS assay successfully quantified PBD from NNAL-induced protein adducts.
- The method demonstrated high sensitivity and specificity for detecting NNAL bioactivation.
- The assay is applicable to diverse biological matrices, facilitating clinical research.
Conclusions:
- This study presents a viable method for detecting NNAL-induced protein adducts in clinical settings.
- The developed assay is critical for assessing the physiological relevance of NNAL bioactivation and lung cancer risk.
- This advancement supports biomarker development for tobacco exposure and related carcinogenesis.
